Bucuresti Vol2018

SCIENTIFIC PAPERS
SERIES B. HORTICULTURE
Volume LXII, 2018
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University of Agronomic Sciences
and Veterinary Medicine of Bucharest
Faculty of Horticulture
SCIENTIFIC PAPERS
SERIES B. HORTICULTURE
Volume LXII
2018
BucharesT
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SCIENTIFIC COMMITTEE
• Bekir Erol AK - University of Harran, Sanliurfa, Turkey
• Arina ANTOCE - University of Agronomic Sciences and Veterinary Medicine of Bucharest, Romania
• Adrian ASĂNICĂ - University of Agronomic Sciences and Veterinary Medicine of Bucharest, Romania
• Liliana BĂDULESCU - University of Agronomic Sciences and Veterinary Medicine of Bucharest, Romania
• Valentina BOZHKOVA - Fruit Growing Institute Plovdiv, Bulgaria
• Dorin CAMEN - Banat's University of Agricultural Sciences and Veterinary Medicine "King Mihai I of Romania" from
Timisoara, Romania
• Daniela CICHI - University of Craiova, Romania
• Mirela CORDEA - University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca, Romania
• Sina COSMULESCU - University of Craiova, Romania
• Miljan CVETKOVIC, University of Banja Luka, Bosnia and Herzegovina
• Elena DELIAN - University of Agronomic Sciences and Veterinary Medicine of Bucharest, Romania
• Alin DOBREI - Banat University of Agricultural Sciences and Veterinary Medicine Timisoara, Romania
• Elisabeta DOBRESCU - University of Agronomic Sciences and Veterinary Medicine of Bucharest, Romania
• Lucia DRAGHIA - University of Agricultural Sciences and Veterinary Medicine Iasi, Romania
• Franco FAMIANI – Universita degli Studi di Perugia, Italy
• Gheorghe GLĂMAN - President of the Romanian Horticultural Society, Romania
• Luca Corelli GRAPPADELLI - University of Bologna, Italy
• Dorel HOZA - University of Agronomic Sciences and Veterinary Medicine of Bucharest, Romania
• Károly HROTKÓ - Corvinus University of Budapest, Hungary
• Olimpia IORDĂNESCU - Banat University of Agricultural Sciences and Veterinary Medicine Timisoara, Romania
• Karsten KLOPP - Obstbauversuchsanstalt Jork, Germany
• Carmen MANOLE - University of Agronomic Sciences and Veterinary Medicine of Bucharest, Romania
• Mekjell MELAND - Norwegian Institute of Agricultural and Environmental Research - Bioforsk, Norway
• Liu MENGJUN – Hebei Agricultural University, China
• Viorel MITRE - University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca, Romania
• Adrian PETICILĂ - University of Agronomic Sciences and Veterinary Medicine of Bucharest, Romania
• Peter ROUSSOS, Agricultural University of Athens, Greece
• Silviero SANSAVINI - University of Bologna, Italy
• Alvaro STANDARDI - University of Perugia, Italy
• Radu SESTRAŞ - University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca, Romania
• Florin STĂNICĂ - University of Agronomic Sciences and Veterinary Medicine of Bucharest, Romania
• Dorin SUMEDREA - Research Institute for Fruit Growing Pitesti - Maracineni
• Florin TOMA - University of Agronomic Sciences and Veterinary Medicine of Bucharest, Romania
• Valerica TUDOR - University of Agronomic Sciences and Veterinary Medicine of Bucharest, Romania
• Cristos XILOYANNIS - University of Pisa, Italy
EDITORIAL BOARD
General Editor: Dorel HOZA
Executive Editor: Adrian ASĂNICĂ
Members: Bekir Erol AK, Valentina BOZHKOVA, Luca CORELLI GRAPPADELLI, Elena DELIAN,
Elisabeta DOBRESCU, Károly HROTKÓ, Karsten KLOPP, Adrian PETICILĂ, Florin TOMA
PUBLISHERS:
University of Agronomic Sciences and Veterinary Medicine of Bucharest - Faculty of Horticulture
Address: 59 Marasti, District 1, 011464 Bucharest, Romania
E-mail: [email protected], Webpage: www.horticultura-bucuresti.ro
CERES Publishing House

Address: 29 Oastei Street, District l, Bucharest, Romania
Phone: + 40 317 90 23, E-mail: [email protected], Webpage: www.editura-ceres.ro
Copyright 2018
To be cited: Scientific Papers. Series B. Horticulture, Vol. LXII, 2018
The publishers are not responsible for the opinions published in the Volume. They represent the authors’ point of view.
Print ISSN 2285-5653, CD-ROM ISSN 2285-5661, Online ISSN 2286-1580, ISSN-L 2285-5653
International Database Indexing: Web of Science Core Collection (Emerging Sources Citation Index), Index
Copernicus, CABI, Ulrich’s Periodicals Directory (ProQuest), PBN, Scientific Indexing Service, Cite Factor
(Academic Scientific Journals), Scipio, OCLC (WorldCat), Research Bible, Google Scholar.
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SUMMARY
FRUIT GROWING
1 EFFECT OF INFECTION BY ILAR-VIRUSES ON VEGETATIVE GROWTH OF

SWEET CHERRY IN NURSERY AND YOUNG TREES - Aneliya BORISOVA... 17
2 MECHANICAL PROPERTIES OF THE FRUITS OF SOME PERSPECTIVE
SWEET CHERRY CULTIVARS - Simeon KRUMOV, Nicolai CHRISTOV ….... 21
3 RESEARCH ON THE VARIETY INFLUENCE ON VEGETABLE GROWTH AT
APRICOT SPECIES IN SANDY SOILS CONDITIONS IN SOUTHERN
OLTENIA - Viorel ENACHE, Mihaela CROITORU, Milica DIMA …….……. 25
4 STUDIES REGARDING THE INFLUENCE OF PRE-HARVEST AND POST-
HARVEST TREATMENTS UPON THE QUALITY OF SOME APPLE FRUIT
VARIETIES - Adrian CHIRA, Lenuța CHIRA, Elena DELIAN, Constanța
ALEXE ………………………………………………………………………………. 29
5 SYSTEMS FOR SHAPING AND PRUNING OF PLUM TREES USED IN
BULGARIA - Denitsa Serbezova ………………………………………………… 33
6 ASPECTS REGARDING THE QUALITY EVOLUTION OF SOME APRICOT
FRUIT VARIETIES DEPENDING ON THE STORAGE CONDITIONS - Lenuța
CHIRA, Adrian CHIRA, Elena DELIAN, Ligia ION, Constanța ALEXE 39
7 ECONOMIC EFFECT OF SUSTAINABLE APPLE PRODUCTION - Iliyana
Krishkova, Denitsa Serbezova …………………………………………………… 45
8 STUDIES ON THE BEHAVIOR OF SOME ELITE AND VARIETIES OF PEAR
IN THE POLLINATION PROCESS - Mihaela (ERCULESCU) STOICA, Dorel
HOZA, Lenuţa Constantina CHIRA, Constantin PĂUN, Ligia ION …………. 51
9 RESEARCH ON THE ISOLATION OF GENOMIC DNA FROM OLD APPLE
VARIETIES - Ligia ION ……………………………………………………..…..… 55
10 EFFECTS OF GRAFTING COMBINATIONS ON THE FRUIT QUALITY FOR
THE PINOVA APPLE TREE - Radu BĂRĂSCU, Dorel HOZA, Ligia ION,
Gheorghița HOZA ……………………..…………………………………………. 59
11 PHYSICAL AND CHEMICAL PARAMETERS OF THE FRUIT IN FOUR
PRUNUS DOMESTICA LOCAL POPULATIONS FROM BUZAU COUNTY -
Daniel Constantin POTOR, Aurora DOBRIN, Mihaela Ioana GEORGESCU,
Dorel HOZA ……………………………………………………………….……… 65
12 ˈHARLAYNEˈ Х ˈHARCOTˈ – PERSPECTIVE CROSSBREED FOR COMBI-
NING GOOD FRUIT QUALITY AND RESISTANCE TO PLUM POX VIRUS -
Marieta NESHEVA, Valentina BOZHKOVA, Snezhana MILUSHEVA 71
13 EVALUATION OF INTRODUCED PLUM CULTIVARS UNDER
AGROCLIMATIC CONDITIONS OF PLOVDIV REGION, BULGARIA -
Valentina BOZHKOVA, Petar SAVOV ………………………………………… 77
14 ADVENTITIOUS SHOOT REGENERATION FROM PETIOLE EXPLANTS IN
BLACK CHOKEBERRY (ARONIA MELANOCARPA) - Ionela RUSEA, Aurel
POPESCU, Valentina ISAC, Anca Nicoleta ȘUȚAN, Dorel HOZA ………….. 83
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15 PHYSIOLOGICAL STUDY OF VARIOUS CULTIVARS OF PUNICA
GRANATUM (L.) WITH AN EYE TO ACCLIMATIZATION TO ROMANIA’S
ENVIRONMENTAL CONDITIONS - Ana-Maria PETRE, Aurelia DOBRESCU,
Adrian PETICILĂ, Monica Luminița BADEA ………………………………… 93
16 EFFECT OF DIFFERENT AMINO ACID FOLIAR FERTILIZERS ON YIELD
AND FRUIT QUALITY OF ”REDIX” APPLE CULTIVAR - Alina Viorica ILIE,
Cristina PETRIȘOR, Dorel HOZA …………………………………………...… 101
17 RESEARCHES ON GENETIC RESISTANCE TO APRICOT VARIETY
BERGERON TO FROST FROM WINTER - Constantin PĂUN ……………...… 105
18 STUDY OF THE CYTOLOGICAL CHARACTERISTICS AND GERMINATION
POTENTIAL OF SOME PEACH CULTIVARS FROM RSFG CONSTANTA -
Corina GAVĂȚ, Elena DOROFTEI, Liliana MIRON, Vlăduț Alexandru
OPRIȚĂ, Georgeta BELENIUC ……………………………………………..…. 109
19 BEHAVIOR OF SOME APRICOT CULTIVARS GRAFTED ON NEW
VEGETATIVE ROOTSTOCKS - Vlăduț Alexandru OPRIȚĂ, Corina GAVĂȚ 115
20 PRODUCTIVITY OF GOOSEBERRY VARIETIES IN THE REPUBLIC OF
MOLDOVA CONDITIONS - Parascovia SAVA …………………………...…….. 119
21 STUDIES ON IN VITRO FERTILIZATION BIOLOGY OF MESPILUS
GERMANICA L. CV. İSTANBUL - Sultan Filiz GÜÇLÜ, Hasan BEKTAŞ,
Fatma KOYUNCU ………………………..……………………………………….. 123
22 RELATIONSHIP BETWEEN VEGETATIVE GROWTH AND NUT
CHARACTERISTICS IN ALTERNATE BEARING PISTACHIO (PISTACIA
VERA) CULTIVARS EXPOSED TO DROUGHT - Hakan CETINKAYA,
Muhittin KULAK, Ahsen Işık OZGUVEN …………..…………………...……… 127
23 EVALUATION OF POMOLOGICAL PROPERTIES OF SOME ORANGE
VARIETIES FROM WEST MEDITERRANEAN, TURKEY - Gulay DEMIR
Fatma KOYUNCU ………………………….………………………..…………… 135
24 POLLEN PERFORMANCES OF NATURALLY GROWN BLACKBERRIES IN
ISPARTA-TURKEY - Sultan Filiz GÜÇLÜ, Ayşe Gül SARIKAYA, Fatma
K O Y U N C U ............................................................................................................ 141
25 EFFECT of POSTHARVEST OXALIC ACID TREATMENT ON COLD
STORAGE OF APRICOT CV. APRIKOZ - Mehmet Ali KOYUNCU, Tuba
SECMEN, Cemile Ebru ONURSAL, Derya ERBAS, Atakan GUNEYLI, Seda
SEVINC UZUMCU, Fatma KOYUNCU ……………….…………………….… 147
26 EFFECTS OF PRE-HARVEST RETAIN TREATMENTS WITH MAP ON COLD
STORAGE QUALITY OF SWEET CHERRY CV. 0900 ZIRAAT - Atakan
GUNEYLI, Mehmet Ali KOYUNCU, Cemile Ebru ONURSAL, Derya ERBAS,
Melike CETİNBAS, Sinan BUTAR, Fatma KOYUNCU …………………..….. 153
27 AN INVESTIGATION ON EFFECTS OF DRY AND WET CLIMATE
CONDITIONS ON PISTACHIO (PISTACIA VERA) YIELD IN MIDDLE
EUPHRATES BASIN SOUTHEAST OF TURKEY - Mehmet Ali ÇELİK,
Muhittin KULAK, Hakan CETİNKAYA, Mehmet KOC, Ali GÖCERİ, Salman
ÖZÜPEKÇE ……………………………………………………………………..…. 159
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28 LOW COST AND LABORATORY SCALE NIR SPECTROSCOPY FOR
QUALITY EVALUATION OF FRUITS AND VEGETABLES - Zehan
KESİLMİŞ, Kubilay K. VURSAVUŞ ……………………………………...……. 165
29 INCREASING THE ECONOMIC PERFORMANCE BY PROMOTING HIGH-
DENSITY APPLE ORCHARDS IN THE DÂMBOVIŢA FRUIT BASIN -
Gheorghe PETRE, Valeria PETRE, Adrian ASANICA ……………….………. 169
30 TWENTY YEARS OF EXPERIENCE IN INTENSIVE PLUM PRODUCTION ON
PRUNUS CERASIFERA EHR. ROOTSTOCK - Miljan CVETKOVIĆ, Nikola
MICIC ………………………………………………………………………….……. 175
31 SENSORIAL EVALUATION OF 26 HIGHBUSH BLUEBERRY VARIETIES IN
ROMANIA - Adrian ASĂNICĂ ………………………………………………….. 181
32 EVALUATION OF THE CONSUMER PREFERENCE FOR SWEET CHERRY
FRUITS AT THE “SWEET CHERRY FEST” IN ISTRITA-BUZAU - Adrian
ASĂNICĂ, Florin STĂNICĂ, Valerica TUDOR, Alexandru IACOB, Violeta
ZOLOTOI, Livia PEROJUC ………………………………………………….…. 187
33 THE BEHAVIOR OF GALA, JONAGOLD, GOLDEN DELICIOUS AND
GRANNY SMITH APPLE VARIETIES IN ORGANIC FARMING SYSTEM -
Andreea BUJDEI, Roxana CICEOI, Florin STĂNICĂ ……………………..… 191
34 SOME RESULTS ON NUTRITIONAL PROPERTIES OF ORGANIC ROSE
PETALS AND RELATED PROCESSED PRODUCTS - Ana Cornelia
BUTCARU, Florin STĂNICĂ, Aurora DOBRIN ………………………….…… 197
35 CULTIVATION TECHNOLOGY OF ORGANIC ROSES FOR PETAL
PRODUCTION - Ana Cornelia BUTCARU, Florin STĂNICĂ ………………….. 203
36 MORUS SPP. FOR REVIGORATING SILKWORM BREEDING IN ROMANIA
AND PROMOTING HEALTH BENEFITS OF LEAVES AND FRUITS - Otilia
BOBIS, Daniel Severus DEZMIREAN, Liviu Alexandru MĂRGHITAŞ,
Victoriţa BONTA, Adriana URCAN, Claudia PAŞCA, Adela Ramona MOISE 211
37 IN VITRO EFFECT OF GENOTYPE, GROWTH SEASON AND CYTOKININES
ON PEACH VARIETIES (Prunus persica (L.) Batsch) PROPAGATION - Nazar
AL GHASHEEM, Florin STĂNICĂ, Adrian George PETICILĂ, Oana VENAT 217
38 IN VITRO EFFECT OF VARIOUS STERILIZATION TECHNIQUES ON PEACH
(Prunus persica (L.) Batsch) EXPLANTS - Nazar AL GHASHEEM, Florin
STĂNICĂ, Adrian George PETICILĂ, Oana VENAT ……………..…….…… 227
39 SENSORY ANALYSIS OF THE DEHYDRATED PRODUCT OBTAINED FROM
APPLES HARVESTED FROM THE TRADITIONAL ORCHARDS OF THE
BRAN - ZĂRNEȘTI AREA - Cătălina ITIGAN, Florin STĂNICĂ ……..…….. 235
40 CUTTINGS PROPAGATION OF SOME FIG GENOTYPES (FICUS CARICA) -
Holia AHMAD, Florin STĂNICĂ, Vlad STANCIU …………………………… 241
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VITICULTURE AND OENOLOGY
1. CHARACTERIZATION OF GRAPE AND WINE QUALITY INFLUENCED BY

TERROIR IN DIFFERENT ECOSYSTEMS FROM ROMANIA CULTIVATED
WITH FETEASCĂ NEAGRĂ - Alin DOBREI, Alina Georgeta DOBREI,
Eleonora NISTOR, Gheorghe POȘTA, Mihaela MĂLĂESCU, Marius BALINT 247
2. STUDY OF THE ANTHOCYANIC POTENTIAL OF GRAPES VARIETIES FOR
RED WINES IN DRANIC WINE CENTER - Camelia MUNTEAN, Felicia
STOICA, Constantin BĂDUCĂ CÂMPEANU, Daniela Doloris CICHI ……... 255
3. THE BEHAVIOR OF THREE CABERNET SAUVIGNON CLONES IN VALEA
CALUGĂREASCĂ AREA - Vlad-Andrei FILIP, Elena BRÎNDUȘE, Marian
ION, Petruța Mihaela MATEI, Laura Lidia CĂLUGĂRU …………………… 261
4. TEMPERATURE AND RAINFALL INFLUENCE ON SHOOT LENGTH IN
PINOT NOIR, MERLOT AND CABERNET SAUVIGNON VARIETIES -
Eleonora NISTOR, Alina DOBREI, Alin DOBREI, Dorin CAMEN, Giovan
Battista MATTI …………………………………………………………………… 267
5. VOLATILE PROFILE OF FETEASCA NEAGRA WINES OBTAINED IN
MURFTALAR REGION AND THE INFLUENCE OF THE ORGANIC AND
CLUSTER THINNING VITICULTURAL PRACTICES - Victoria ARTEM,
Arina Oana ANTOCE ………………………………………………….………… 275
6. RESEARCHES REGARDING QUALITY AND QUANTITATIV PERFOR-
MANCE OF SOME TABLE GRAPES IN THE EXPERIMENTAL FIELD OF
UASVM BUCHAREST - Marinela Vicuţa STROE, Daniel Nicolae COJANU 281
7. INFLUENCES OF GRAPE CULTIVATION TECHNOLOGY ON CERTAIN
AROMA COMPOUNDS SIGNIFICANT FOR THE DIFFERENTIATION OF
CABERNET SAUVIGNON AND FETEASCĂ NEAGRĂ WINES - George
Adrian COJOCARU, Arina Oana ANTOCE …………………………………. 287
8. INCREASE OF ANTHOCYANINS ACCUMULATION BY PREHARVEST 24-
EPIBRASSINOLID (24-EBL) APPLICATIONS IN HOROZ KARASI GRAPE
CULTIVAR - Zehra BABALIK, Tunhan DEMİRCİ, Özlem ARAS AŞCI,
Nilgün GÖKTÜRK BAYDAR ………………………………………………….. 295
9. THE INFLUENCE OF PREHARVEST BORIC ACID APPLICATIONS ON THE
ACCUMULATION OF SOME ANTIOXIDANT COMPONENTS IN ALPHONSE
LAVALLÉE GRAPE CULTIVAR - Zehra BABALIK, Tunhan DEMİRCİ,
Özlem ARAS AŞCI, Nilgün GÖKTÜRK BAYDAR ………..…………………… 299
10. MOLECULAR DETECTION OF BLACK ASPERGILLUS AND PENICILLIUM
SPECIES FROM DEALU MARE VINEYARD - Camelia Filofteia DIGUȚĂ,
Radu Cristian TOMA, Călina Petruța CORNEA, Florentina MATEI 305
11. INFLORESCENCE AND FLOWER DIFFERENTIATION IN THE GRAPEVINE
(V. VINIFERA L.) VARIETIES ’ŽILAVKA’ AND ’BLATINA’ - Tatjana
JOVANOVIC-CVETKOVIC, Nikola MICIC, Gordana ĐURIĆ, Miljan
CVETKOVIĆ ………………………………………..…………………..… 311
12. DETERMINATION OF THE PHENOLIC COMPOUNDS, ANTIOXIDANT AND
ANTIRADICAL ACTIVITIES OF SENIRKENT KARASI GRAPE CULTIVAR’S
SKIN AND SEEDS - Filiz HALLAÇ TÜRK, E. Sema ÇETİN, Zehra
BABALIK, Nilgün GÖKTÜRK BAYDAR ……….………………………… 317
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13. STUDY ON THE BEHAVIOR OF THE VICTORIA TABLE GRAPE VARIETY
IN THE HINOVA VITICULTURAL AREA, MEHEDINTI COUNTY - Liviu
Teodor MUNTEAN ……………………………………………………………..….. 323
14. CONSIDERATIONS REGARDING THE USE OF ANTI-HAIL NETS FOR THE
PROTECTION OF TABLE GRAPE PLANTATIONS IN HINOVA
VITICULTURAL AREA, MEHEDINTI COUNTY - Liviu Teodor MUNTEAN … 327
15. EVALUATING SOME OLFACTORY CHARACTERISTICS OF WINE IN THE
CONTEXT OF GLOBAL WARMING IN THE “PLAIURILE DRÂNCEI”
VITICULTURAL REGION - Alina-Mihaela NISTOR, Liviu-Gabriel
GRIGORICĂ, Cristina-Elena SCUTARAȘU, Ștefan-Dragoș COTAN, Valeriu
V. COTEA, Marius NICULAUA ……………………………..………..…………. 331
VEGETABLE GROWING
1 HERITABILITY OF THE TOMATO GENOTYPES RESISTANCE TO THE

HIGH AIR TEMPERATURES - Nadejda MIHNEA, Galina LUPAȘCU ……... 339
2 ESTIMATION OF VARIABILITY PARAMETERS OF SOME QUALITATIVES
COMPONENTS AT A SET OF SWEET CORN LINES FROM TURDA
AGRICULTURAL RESEARCH STATION - Luana PĂCURAR (GRECU),
Alexandru Ioan APAHIDEAN, Gheorghiţa HOZA, Maria DINU, Rodica
SOARE, Maria APAHIDEAN, Voichiţa HAŞ …………………………………. 345
3 RESEARCH REGARDING THE INFLUENCE OF FERTILISER RATES ON
MORPHOLOGICAL FEATURES IN COMMON BEAN (PHASEOLUS
VULGARIS L. CONVAR. VULGARIS) PODS CULTIVATED IN SOLARIUM
AT THE DIDACTIC AND RESEARCH BASE IN TIMISOARA. ROMANIA -
Marius BALINT, Alin DOBREI, Alina Georgeta DOBREI, Gheorghe POȘTA 351
4 THE EFFECTS OF DIFFERENT NITROGEN DOSES ON YIELD AND
NUTRIENT UPTAKE OF ROCKET (ERUCA SATİVA) PLANT - Volkan
YORUK, Hasan DURUKAN, Handan SARAÇ, Ahmet DEMIRBAS, Tolga
KARAKOY ………………………………………………………………..……… 359
5 THE IMPACT OF DIFFERENT METHODS USED FOR IMPROVING FLOWER
BIND UPON THE YIELD OF SOME TOMATO HYBRIDS CULTIVATED ON
MINERAL WOOL SUBSTRATE IN INDUSTRIAL GREENHOUSES -
Alexandra BECHERESCU, Gheorghiţa HOZA, Maria DINU, Olimpia
IORDĂNESCU, Ioan SĂRAC, Daniel POPA ………………………………….. 367
6 RESEARCH ON THE EFFECT OF CLIMATE CHANGE ON THE
PRODUCTION CAPACITY OF SOME SWEET POTATOES VARIETIES
(IPOMOEA BATATAS) CULTIVATED ON PSAMOSOILS IN THE SOUTHERN
AREA OF ROMANIA - Aurelia DIACONU, Alina-Nicoleta PARASCHIV, Reta
DRĂGHICI, Mihaela CROITORU, Milica DIMA ……………………………… 379
7 THE INFLUENCE OF FOLIAR BIOACTIV TREATMENTS ON TOMATOES
SEEDLINGS - Elena DOBRIN, Evelina GHERGHINA, Gabriela LUŢĂ,
Daniela BĂLAN …………………………….………………………….………..… 385
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8 INTER-SPECIFIC (CAPSICUM CHACOENSE HUNZ. AND CAPSICUM
ANNUUM L.) INHERITANCE OF FRUIT DETACHMENT FORCE TRAIT IN
HOT PEPPER - Bekir Bülent ARPACI, Faika YARALI KARAKAN ………… 391
9 ASSESSING TOMATO GERMPLASM FROM V.R.D.S. BUZAU TO IDENTIFY
GENOTYPES WITH DISTINCT FEATURES - Bianca ZAMFIR, Dorel HOZA,
Costel VÎNĂTORU, Camelia BRATU, Elena BARCANU ………………………. 395
10 NEW EGGPLANT GENOTYPES WITH DISTINCT PHENOTYPIC
EXPRESSIVITY OBTAINED AT V.R.D.S BUZĂU - Camelia BRATU, Florin
STĂNICĂ, Costel VÎNĂTORU, Bianca ZAMFIR, Elena BARCANU ………… 399
11 STUDY OF THE EFFECT OF ULTRASOUND ON VEGETABLE CROPS IN
DIFFERENT EXPOSURES - Vangel CHILINGIROV, Manol DALLEV,
Nikolina SHOPOVA, Irena DAKOVA, Galya HRISTOVA, Ivan MITKOV … 403
12 DETERMINATION OF CHLORINE CONCENTRATION AND PRODUCTIVITY
IN SOME BEAN GENOTYPES, FROM NORTH-EAST OF ROMANIA, UNDER
SALT STRESS - Beatrice Alexandra MODIGA, Carmenica Doina
JITĂREANU, Cristina SLABU, Alina Elena MARTA, Mihaela COVAȘĂ ……. 409
13 STUDY OF THE INFLUENCE OF A BIOSTIMULATOR USED IN THE
TREATMENT SEED OF FAMILY APIACEAE BY ULTRASOUND - Irena
DAKOVA, Nikolina SHOPOVA, Manol DALLEV, Vangel CHILINGIROV,
Galya HRISTOVA, Ivan MITKOV …………………………………………...… 417
14 EFFECT OF FOLIAR BIOACTIVE TREATMENTS ON THE OXIDATIVE
STRESS TOLERANCE IN TOMATO SEEDLINGS - Gabriela LUŢĂ, Daniela
BĂLAN, Evelina GHERGHINA, Elena DOBRIN ……………………………... 423
15 DETERMINATION OF THE RESPONSES OF DIFFERENT TOMATO SPECIES
TO TUTA ABSULUTA - Ali KAYAHAN, Betül SIMSEK, Ismail KARACA,
Hakan AKTAS …………………………………………..………….......................... 431
16 RESPONSES ABOUT SEED FORMATION AND SEED PRODUCTION IN
DIFFERENT GENOTYPES OF CAPE GOOSEBERRY (PHYSALIS
PERUVIANA L.) - Nikolay PANAYOTOV ……………………………………....... 437
17 NEW BRED CULTIVARS OF BELL PEPPER OBTAINED AT V.R.D.S. BUZĂU
- Elena BARCANU-TUDOR, Elena Maria DRĂGHICI ……………………….. 445
18 PRELIMINARY STUDY RELATED ON YIELD AND QUALITY POTENTIAL
OF TWO NEW SWEET PEPPERS VARIETIES OBTAINED AT V.R.D.S.
BUZĂU - Elena BARCANU-TUDOR, Elena Maria DRĂGHICI, Aurora
DOBRIN, Andreea STAN, Carmen CONSTANTIN, Mihaela ZUGRAVU,
Oana BUJOR-NENIȚĂ …..………………………………………..……………...... 451
19 INFLUENCE OF PLANT MANAGEMENT SYSTEMS ON GROWTH AND
FRUCTIFICATION OF TOMATO PLANTS IN PROTECTED CULTURE -
Gheorghița HOZA, Maria DINU, Rodica SOARE, Alexandra Dida
BECHERESCU, Ionuț Alexandru APAHIDEAN, Dorel HOZA ……………… 457
20 EFFECT OF GRAFTING ON YIELD AND FRUIT QUALITY OF PEPPER
(CAPSICUM ANNUUM L.) GROWN UNDER OPEN FIELD CONDITIONS -
Volkan ERGUN, Hakan AKTAS …….……………………………………….…... 463
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21 RESEARCH ON THE EFFECT OF VERMICOMPOST FERTILIZATION ON
EGGPLANTS SEEDLINGS (SOLANUM MELONGENA L.) - Constantin PETRE,
Elena DOBRIN, Elena Maria DRAGHICI …..……………………….… 467
22 EFFECT OF THE USE OF NEW METHODS FOR THE REMEDIATION OF OIL
POLLUTED SOIL - Elena Maria DRAGHICI, Virgil SCARLAT, Maria PELE,
Elena DOBRIN, Gabi-Mirela MATEI, Sorin MATEI …………...…………...... 471
23 THE INFLUENCE OF ALTERNATIVE TECHNOLOGICAL SEQUENCES ON
THE QUALITY OF MELON PRODUCTION - Rodica SOARE, Maria DINU,
Cristina BĂBEANU, Marin SOARE ………………………………………..……... 477
24 EFFICACY OF PLANT BASED BIOPRODUCTS AGAINST TOMATO
SEEDLINGS DAMPING OFF DISEASE - SHORT OVERVIEW - Steliana
RODINO, Marian BUTU, Alina BUTU …………………………………….……... 483
25 DETERMINATION OF STRENGTH PROPERTIES FOR MECHANICAL
HARVEST OF PARSLEY (PETROSELINUM CRISPUM) - Mehmet Emin
GOKDUMAN, Deniz YILMAZ ………………………………….………..………. 487
26 WHICH IS CRUCIAL FOR HETEROSIS? TRAITS, GENETIC OR
CHARACTERISTIC DIVERSITY: PUNGENCY PARADIGM - Bekir Bülent
ARPACI, Cihan FIRAT, Mehmet KOÇ, Faika YARALI KARAKAN ……..…. 491
27 THE EFFICACY OF DIFFERENT TREATMENTS FOR PATHOGENS CON-
TROL ON THE EGGPLANT CROPS IN THE FIELD - Mihaela Alina BUZATU,
Marcel COSTACHE, Dorel HOZA, Gabriela ȘOVĂREL, Stelica CRISTEA …. 495
28 PATHOGENS WITH ECONOMIC IMPORTANCE FOR TOMATO CROPS
GROWING IN THE FIELD AND THEIR CONTROL - Iuliana MÂNDRU,
Marcel COSTACHE, Dorel HOZA, Stelica CRISTEA …………………………... 499
29 EXOGENOUS CYTOKININ APPLICATION INCREASED THE CAPSAICIN
AND ASCORBIC ACID CONTENT IN PEPPER FRUIT - Mostafakamal
SHAMS, Ertan YILDIRIM, Melek EKINCI, Guleray AGAR, Metin TURAN,
Raziye KUL …………………………………………..…...………………..………... 507
30 ESSENTIAL OILS AND HOMEMADE FUNGICIDES AGAINST FUSARIUM
OXYSPORUM F.SP. LYCOPERSICI TOMATO PATHOGEN – Andreea-Alexandra
TIMOFTE, Andreea – Mihaela FLOREA, Florin Daniel LIPȘA, Eugen ULEA … 513
31 POSITIVE ASPECTS OF AN ORNAMENTAL VEGETABLE GARDEN AND
ITS EFFECTS TOWARDS FAMILY AND COMMUNITY SUSTAINABILITY -
Florina-Maria GALEA (DELEANU), Neculai MUNTEANU, Vasile STOLERU,
Gabriel Ciprian TELIBAN ……………………………….……………………..…. 517
FLORICULTURE, ORNAMENTAL PLANTS

DESIGN AND LANDSCAPE ARCHITECTURE
1 PRELIMINARY RESEARCHON THE INFLUENCE OF THE PLANTING TIME

ON THE GROWING AND BLOOMING OF VARIETY OF HERBACEOUS
PEONY IN THE FIELD - George Nicolae CAZAN, Sorina PETRA, Florin
TOMA …………………………………………………………………..………… 523
11
2 PRELIMINARY RESEARCH ON THE GROWTH AND FLOWERING OF
DIFFERENT VARIETIES OF HERBACEOUS PEONY UNDER CONDITIONS
OF CONTAINERIZED AND FORCED CULTIVATION - George Nicolae
CAZAN, Sorina PETRA, Florin TOMA ……………………………………….. 533
3 TREE SPECIES SELECTION GUIDELINES FROM THE PERSPECTIVE OF
BIOCLIMATIC LANDSCAPE DESIGN - Vladimir Ionuț BOC ……………..….. 541
4 ANALYSIS OF CHANGES IN GROWING DEGREE-DAY VALUES BY
ALTITUDE: OIL ROSE (ROSA DAMASCENA MILL.) CASE - Ali YUCEL,
Atılgan ATILGAN, Cagatay TANRIVERDI, Hasan ÖZ ……...………………. 551
5 IMPROVING BIOLOGICAL AND PHYSIOLOGICAL PARAMETERS OF
NERIUM OLEANDER L. CUTTINGS BY USING BIOSTIMULATING
SUBSTANCES - Diana VÂȘCĂ-ZAMFIR, Elena DELIAN, Ramona COTRUȚ 559
6 EFFECT OF RHIZOGENIC BIOSTIMULATORS ON ROSMARINUS
OFFICINALIS ROOTED CUTTINGS BIOCHEMICAL COMPOSITION - Diana
VÂȘCĂ-ZAMFIR, Daniela BĂLAN, Gabriela LUȚĂ, Evelina GHERGHINA 565
7 COPPER EFFECT ON SEED GERMINATION AND PLANT SPROUTING OF
ALYSSUM MURALE SPECIES - Maria BRÎNZĂ, Elena Liliana CHELARIU,
Andreea AIORDĂCHIAEI Lucia DRAGHIA ……………………….…………. 571
8 DECODING THE SIGNIFICANCES OF BUILT LANDSCAPE PROFILES -
Doina Mira DASCĂLU, Mirela COJOCARIU …………………………….….. 575
9 STUDIES REGARDING THE BEHAVIOUR OF ORNAMENTAL SPECIES
LAGURUS OVATUS IN CROPPING CONDITIONS FROM N-E AREA OF
ROMANIA - Elena Liliana CHELARIU, Lucia DRAGHIA, Maria BRÎNZĂ,
Mirela COJOCARIU, Bogdan-Vlad AVARVAREI ………………………….. 581
10 THE RESPONSE OF BULGARIAN SPRAY-CARNATION (D.
CARYOPHYLLUS F. SPRAY, HORT) CV. RUSALKA TO DROUGHT - IN
VITRO INDUCED BY DIFFERENT PEG CONCENTRATIONS - Violeta
IVANOVA, Nadejda ZAPRIANOVA, Bistra ATANASOVA, Valentin
PANCHEV ……………………………………………………………..….……… 585
11 ASPECTS REGARDING THE ORNAMENTAL VALUE OF SOME ROSE
NURSEY SPONTANEOUS IN IASI DISTRICT CONDITIONS - Roberto
BERNARDIS, Marius DASCĂLU ……………………………………………… 591
12 STUDIES ON GROWTH AND DEVELOPMENT OF HEDERA HELIX L. ON
DIFFERENT WOODY SPECIES - Cristina Rodica MĂNESCU, Claudiu
BUDA, Florin TOMA, Sorina PETRA ……………………………………… 597
13 POLLEN ANALYSIS IN SOME TULIP CULTIVARS - Mirela Irina CORDEA,
Ioana Delia POP, Sonia BORS-OPRIŞA, Lucia MIHALESCU ……………. 601
14 THE EFFECT OF DIFFERENT LIGHT CONDITIONS ON THE GROWTH AND
DEVELOPMENT OF CHLOROPHYTUM AMANIENSE ENGL. ‘FIRE FLASH’
- Manuela MANDA, Carmen NICU, Diana ZAMFIR-VÂȘCĂ ……….……... 605
15 TINERETULUI PARK – FUNCTIONAL AND COMPOSITIONAL
COMPONENTS IN THE URBAN LANDSCAPE OF BUCHAREST - Anca
STĂNESCU, Suzana COCIOABĂ ……………………………………….……... 611
12
16 NATURAL HERITAGE CONSERVATION IN BUILT-UP AREAS. STUDY ON
THE CIRCUS PARK AS AN URBAN LANDSCAPE ASSET - Suzana
COCIOABĂ, Anca STĂNESCU ……………..………………………………….. 615
17 LEAF STOMATAL PARAMETERS OF IRIS GERMANICA L. INFLUENCED
BY CULTIVAR AND ARBUSCULAR MYCORRHIZAE INOCULATION IN
FIELD CONDITIONS, ROMANIA - Ioana CRIŞAN, Roxana VIDICAN, Vlad
STOIAN, Sorin VÂTCĂ, Andrei STOIE …………………………………… 619
18 PRELIMINARY TESTING OF SRAP PRIMERS IN FOUR RANUNCULACEAE
SPECIES FROM ROMANIA - Cristina D. KELEMEN, Orsolya BORSAI,
Katalin SZABO, Doina CLAPA, Monica HÂRŢA, Doru PAMFIL ……………. 629
MISCELLANEOUS
1 POSSIBILITIES FOR SMALL-SCALE COMPOSTING OF HORTICULTURAL

PLANT WASTES - Milena YORDANOVA, Erik MINCHEV, Radoslav
BORISOV …………………………………………………………….……………. 637
2 MORPHOLOGICAL AND ANATOMICAL INVESTIGATION OFALOYSIA
CITRODORA PALAU – NEW MEDICINAL PLANT INTRODUCED IN
ROMANIA - Vasilica LUCHIAN, Elena SĂVULESCU, Jorj PĂDURARU,
Vlad POPA, Viorica LAGUNOVSCHI-LUCHIAN ............................................. 643
3 DYNAMICS OF MICROBIOLOGICAL INDICATORS FOR COMPARATIVE
STUDY OF COMPОST VARIANTS - Boyka MALCHEVA, Milena
YORDA NOVA, Ra doslav BOR ISOV , Teodora VICHEVA, May a
NUSTOROVA ……………………………………………….……………….… 649
4 CONTRIBUTIONS TO HALOPHILIC FLORA AND VEGETATION IN
OLTENIA (ROMANIA) - Daniel RĂDUȚOIU, Cătălin George SIMEANU, Ion
STAN ……………………………………………………………….……………… 655
5 PLANT GROWTH PROMOTING BACTERIA WITH ANTIFUNGAL
ACTIVITY - Oana-Alina BOIU-SICUIA, Sorina DINU, Cristina PETRIȘOR,
Florica CONSTANTINESCU………………………………………….…………. 661
6 ANTIMICROBIAL POTENTIAL OF ROMANIAN SPONTANEOUS FLORA -
A MINIREVIEW - Tomina PURCARU, Camelia DIGUŢĂ, Florentina MATEI 667
7 TRICHODERMA SPP. SOURCE OF HYDROLASE ENZYMES WITH ROLE IN
F. OXYSPORUM INHIBITION - Mohammed Naithel RADHI, Cristina
PETRISOR, Florica CONSTANTINESCU, Ioan ROSCA, Hattf Bazool
FARHOOD, Hiba ABD- AL ADHEEM, Gheorghe STOIAN …………………. 681
8 EFFECT OF PERIODS AND REPETITION OF THE BURNING METHOD IN
THE CONTROL OF WATER HYACINTH (EICHHORNIA CRASSPIES)
PLANT - Al-Gburi Omar ABDULRAZZAQ SHIHAP, Talal Abed KHATTAB,
Cristina PETRIȘOR ……………………….……………...……………………... 687
9 BAKERY PRODUCTS FORTIFIED WITH DRIED FRUITS OF ARONIA
MELANOCARPA - Monica CATANĂ, Luminița CATANĂ, Enuța IORGA,
Adrian Constantin ASĂNICĂ, Nastasia BELC ……………………………….. 693
13
10 INTEGRATING SOIL PH MEASUREMENT INTO AN INTERNET OF THINGS
APPLICATION - Adrian ZĂRNESCU, Răzvan UNGURELU, Marius – Ionuţ
MACOVEI, Gaudenţiu VĂRZARU ……………….………………………..….. 703
11 ADVANTAGES AND DISADVANTAGES OF PESTICIDE ANALYSIS
METHODS USED IN AGRICULTURAL SAMPLES - Elena-Cornelia
M I T R A N , E l e n a P e r d u m , O v i d iu G e o r g e I O R D A C H E , I u l i a n a
DUMITRESCU ……………………………………………………………………. 709
12 SCREENING OF MICROORGANISMS DISPLAYING ACETYL XYLAN
ESTERASE ACTIVITY - Aglaia BURLACU, Florentina ISRAEL-ROMING,
Călina Petruța CORNEA ………………………………………………………… 715
14
Fruit
Growing
15
16
Scientific Papers. Series B, Horticulture. Vol. LXII, 2018
EFFECT OF INFECTION BY ILAR-VIRUSES

ON VEGETATIVE GROWTH OF SWEET CHERRY
IN NURSERY AND YOUNG TREES
Aneliya BORISOVA
Institute of Agriculture - Kyustendil, 2500 Kyustendil, Bulgaria
Corresponding author email: [email protected]
Abstract
The effect of infection with Prunus necrotic ringspot virus (PNRSV) and Prune dwarf virus (PDV) on vegetative growth
of sweet cherry trees was investigated in a nursery and during the first three years in an orchard. The subjects of the
research were trees of cultivars ʻVanʼ, ʻKozerskaʼ, ʻDrogans yellowʼ, ʻLambertʼ and ʻStefaniaʼ. In the spring of 2012
virus-free P. mahaleb rootstocks (IK-М9) were budded with virus-free buds of these cultivars. Fourteen days after that,
they were artificially infected with isolates of PNRSV, PDV and mixed infection of them, using 'double chip budding'
method. Serological analyses were made by DAS-ELISA method. There was a significant negative virus effect on the
vegetative growth in most of the single infected with PNRSV or PDV and all mixed infected trees of Van and Lambert in
a nursery. As an average for the period of study there were not significant virus effects on the trunk diameter, the height
of the trees and the length of the shoots of virus infected trees of the cultivars ʻStefaniaʼ, ʻKozerskaʼ and ʻVanʼ,
compared to the healthy young trees.
Key words: influence, PNRSV, PDV, sweet cherry, vegetative growth.
INTRODUCTION combined infections, the damage is always

higher than if the viruses infected separately
The viral infections are one of the main (Cropley, 1968). In the fruit-bearing orchards
problems in the production of healthy these viruses induce also growth and crop yield
propagated material from stone fruit species reduction (Parker et al., 1959; Posnette et al.,
and profitable fruit production. The most 1968). They can delay maturity in fruit
common and economically important among development up to 2 weeks compared to virus-
the viral pathogens infecting the sweet cherry free trees (Howell and Mink, 1984).
trees are Prunus necrotic ringspot virus The purpose of our study was to investigate the
(PNRSV) and Prune dwarf virus (PDV). These influence of PNRSV and PDV on the some
viruses, members of the genus Ilarvirus, spread vegetative properties of five sweet cherry
not only by infected propagating material, but cultivars grafted on Prunus mahaleb rootstock.
also by seed and pollen (Amari et al., 2009;
Aparicio et al., 1999; Mink, 1993). MATERIALS AND METHODS
Quantitative data on the relationship between
virus infection and the effect on the vegetative The investigation was carried out at the
and reproductive development of sweet cherry Institute of Agriculture-Kyustendil, Bulgaria.
trees is limited and most the publications are In the spring of 2012, virus-free P. mahaleb
from the last century. rootstocks (IK-М9) were budded with virus-
PNRSV and PDV may cause a significant free buds of cultivars ʻVanʼ, ʻKozerskaʼ,
reduction in taking of buds, varying by ʻDrogans yellowʼ, ʻLambertʼ and ʻStefaniaʼ. 14
percentages dependent on the rootstocks days after that, they were artificially infected
(Nemeth, 1972; Proebsting et al., 1995). with isolates of PNRSV, PDV and mixed
The infection of Ilarvirus also has an adverse infection of them, using 'double chip budding'
effect on the development of young trees in the method (Table 1). Ten trees of each cultivar
nursery, and retards their growth (Milbrath, were infected with PNRSV, PDV and the
1950; Milbrath, 1957; Nemeth, 1986; Gilmer et combination of both. Virus-free trees from each
al., 1976; Nyland et al., 1976). In the case of cultivar were used as controls.
17
Table 1 Virus treatments and source of inoculum viruses on the vegetative properties of trees of
from commercial sweet cherry orchards investigated five cultivars in a nursery are
Virus treatment Source of virus represented in Table 2.
PNRSV naturally infected cherry, cultivar In most of the single and all mixed infected
‘Merchant’
PDV naturally infected cherry, cultivar trees of cultivar ʻVanʼ and ʻLambertʼ was
‘Superstar’ found a significant negative influence of the
PNRSV+ PDV I naturally infected cherry, cultivar viruses on vegetative growth of tree.
‘Diana’
PNRSV+ PDV II naturally infected cherry, cultivar
The development was the least disturbed by
‘Priusadebnaya’ PNRSV infection: the rootstock diameter was
less by 8.2% in cultivar Lambert and 10.6% in
The 2-year-old nursery trees of cultivars ʻVanʼ, Van, the diameter of scion was less by 17.0%
ʻKozerskaʼ and ʻStefaniaʼ artificially infected in Van, the height of the young tree was less by
with PNRSV, PDV and mix combination, as 2.3% in Lambert and 12.4% in Van.
well as - virus free ones were planted in autumn The strongest effect on the development was
2013 in randomised complete blocks, at a observed in the combined infection of PNRSV
spacing 4.80 х 4.00 m. and PDV: the diameter of rootstocks was less
Each variation of each cultivar was represented by 21.4 - 25.3% in Van, the diameter of scion
by three trees. was less by 21.9 -30.4% in Van, the height of
The performance of these trees in terms of the tree was less by 27.8 - 37.0% in Lambert. In
vegetative growth – trunk diameter, height of the cultivars ʻDrogans yellowʼ, ʻStefaniaʼ and
the tree and length of the shoots on infected and ʻKozerskaʼ the viruses and the mixed infections
virus free trees was examined for three years. of them insignificantly reduced vegetative
In February 2015, a pruning of healthy and properties of the trees.
infected trees was carried out. With the exception of the cultivar ʻKozerskaʼ,
The viral status of the used rootstocks IK-М9 where PNRSV infection induced reduction of
and sweet cherry cultivars was determined by the diameter and the height of trees by 32.5%,
ELISA testing. All plant materials were mixed infection reduced the scion diameter by
subjected to DAS-ELISA using the PNRSV, 24.7%. In cultivar ʻStefaniaʼ the combined
PDV, PPV, CLRV and RpRSV detection kit of infection decreased the diameter of rootstocks
Loewe Biochemica (GmbH, Germany) and by 15.5%, which was statistically proven.
Cocktail-ELISA for identification of ACLSV
following recommendations of the supplier. In a non – fruit - bearing sweet cherry
The infection of virus free trees by pollen orchard:
transfer was followed by regular ELISA testing The effect of virus infections on the vegetative
each year. properties of the investigated cultivars in the
The data was statistically processed by analysis orchard is shown in Table 3. As an average for
of variance using F for test significance and the period of study, there were not significant
LSD for significance of the differences virus effects on the trunk diameter, the height
between variant means and control, at level of the trees and the length of the shoots of virus
P<0.05, 0.01 and 0.001, depending on data infected trees of cultivars ‘Stefania’,
dispersion. ‘Kozerska’ and ‘Van’, compared to the healthy
The data analysis was performed by computer young trees.
programs developed by Maneva, 2007 on the Although it was noted that virus infections
base of standard statistical algorithms suitable insignificantly decreased the trunk diameter by
for small set of data with biological origin 2.2% in trees of cultivar ‘Van’ infected with
(Sokal and Rohle, 1981). PNRSV to 27.8% in trees ‘Stefania’ with
PNRSV+PDV, the height of the young trees by
RESULTS AND DISCUSSIONS 3.8% in cultivar ‘Stefania’ infected with
PNRSV to 13.9% in ‘Stefania’ with PDV, the
In a nursery shoot length by 3.2% in trees of ‘Stefania’ with
The data from the experiment of the influence PNRSV to 28.9% in trees of ‘Kozerska’ with
of PNRSV, PDV and mixed infection with both mix infection PNRSV+PDV.
18
Table 2. Vegetative properties of virus-free and Howell, 2001). It should be noted that in our
virus infected trees in a nursery experiment, it was used only P. mahaleb
Rootstock Scion
rootstock and this may be one of the reasons for
Virus Height of the established insignificant influence of these
Cultivar diameter, diameter,
treatments tree, cm
cm cm viruses on the vegetative growth of infected
virus free 1.82 1.40 146.8 young sweet cherry trees in an orchard.
ʻDrogans PNRSV 1.66 ns 1.28 ns 132.6 ns
yellowʼ PDV 1.76 ns 1.11 ns 126.4 ns
However, recent research revealed that trees on
PNRSV+ 1.82 ns 1.36 ns 134.6 ns some new dwarfing rootstock like Gisela 6,
PDV I
PNRSV+ 1.78 ns 1.32 ns 124.6 ns
Damil, Inmil and others exhibited detrimental
PDV II reactions to infection by PDV and PNRSV
Sd
F
0.1146
0.6574
0.1660
0.9819
17.5374
0.4995
(Lang et al., 1998; Andersone et al., 2002;
LSD 0.05 0.243 0.3518 37.17 Lankes, 2007).
virus free 1.94 1.40 160.4
ʻStefaniaʼ PNRSV 1.84 ns 1.52 ns 157.6 ns
Table 3. Vegetative properties of virus-free and virus
PDV 1.80 ns 1.34 ns 152.8 ns
PNRSV+ 1.64 ⃰ 1.40 ns 149.2 ns infected trees in orchard
PDV I
PNRSV+ 1.96 ns 1.36 ns 164.4 ns
Trunk Height Length of
Cultivar
PDV II Virus diameter, of tree, one-year
Sd 0.1129 9.2518 11,1132 treatments cm cm shoots, cm
F 2.5830 1.1403 0.5873 Average (2014-2016)
LSD 0.05 0.2394 0.1961 23.56 virus free 1.53 228.3 41.89
virus free 1.92 1.78 185.8 PNRSV 1.36 ns 219.6 ns 40.52 ns
ʻKozerskaʼ PNRSV 1.90 ns 1.20 ⃰ ⃰ 125.4 ⃰ ʻStefaniaʼ PDV 1.20 ns 196.6 ns 35.08 ns
PDV 1.76 ns 1.58 ns 182.0 ns
PNRSV+ 1.11 ns 200.3 ns 33.99 ns
PNRSV+ 1.84 ns 1.34 ⃰ 145.0 ns
PDV I
PDV
PNRSV+ 1.86 ns 1.58 ns 180.8 ns Sd 0.2503 33.2811 7.5832
PDV II F 1.0957 0.4180 0.5341
Sd 0.1843 0.1782 22.4815 LSD 0.6133 81.53 18.57
F 0.2285 3.2544 2.9007 0.05
LSD 0.05 0.3906 0.3778 47.66 virus free 1.82 260.6 56.95
ʻKozerskaʼ

ʻLambertʼ PNRSV 2.00 ns 1.48 ns 159.8 ns PDV 1.726 ns 248.0 ns 48.00 ns
PDV 1.84 ⃰ 1.40 ns 143.2 ns
PNRSV+ 1.473 ns 238.3 ns 40.46 ns
PNRSV+ 1.80 ⃰ 1.18 ⃰ 118,0 ⃰
PDV I PDV
PNRSV+ 1.72 ⃰ 1.10 ⃰ 101.8 ⃰ ⃰ Sd 0.3005 19.6718 6.8374
PDV II F 0.7765 0.5017 2.3030
Sd 0.1582 0.1259 16.0780 LSD 0.7362 48.19 16.75
F 2.6772 3.7680 5.5424 0.05
LSD 0.05 0.3354 0.2669 34.08 virus free 1.52 249.6 49.34
ʻVanʼ
ʻVanʼ PNRSV 1.84 ns 1.36 ⃰ 153.8 ns

PDV 1.386 ns 232.3ns 40.03ns
PDV 1.68 ⃰ ⃰ 1.36 ⃰ 145.8 ⃰
PNRSV+ 1.62 ⃰ ⃰ 1.28 ⃰ 129.6 ⃰ ⃰ ⃰
PNRSV+ 1.36 ns 223.3 ns 38.21sn
PDV I PDV
PNRSV+ 1.54 ⃰ ⃰ ⃰ 1.14 ⃰ ⃰ 136.0 ⃰ ⃰ Sd 0.0879 29.7069 5.853
PDVII F 1.5340 0.2808 1.5135
Sd 0.1278 0.1263 11.1178 LSD 0.2155 72.78 14.34
F 5.2043 4.1705 5.1885 0.05
LSD 0.5 0.2709 0.2678 23.56
CONCLUSIONS
During the period of the investigation, each
spring the virus free trees of the investigated There was a significant negative virus effect on
cultivars were retested for the present of viruses the vegetative growth in most of the single
- PNRSV and PDV. The results of the infected with PNRSV or PDV and all mixed
serological analyzes confirmed that all control infected trees of cultivars ‘Van’ and ‘Lambert’
trees were virus free. in a nursery.
Sweet cherry trees in Bulgaria are grown As an average for the period of study there
primarily on Mahaleb (P. mahaleb) or Mazzard were not significant virus effects on the trunk
(Prunus avium) seedlings. These rootstocks are diameter, the height of the trees and the length
generally tolerant to infection by the pollen- of the shoots of virus infected trees of the
borne ilarviruses PDV and PNRSV (Lang and cultivars ‘Stefania’, ‘Kozerska’ and ‘Van’
19
grafted on Prunus mahaleb rootstock, Lang G., Howell W., 2001. Lethal sensitivity of some
compared to the healthy young trees. new cherry rootstocks to pollen-borne viruses. Acta
Hort. 557: 151-154.
Lankes, Ch., 2007. Testing of prunus rootstock clones
ACKNOWLEDGEMENTS for virus tolerance. Acta Hort 732: 351-353
Maneva S., 2007. Mathematical modelling in plant
This article is published with the financial protection. PhD Dissertation, Plant Protection
support of the Ministry of Education and Institute, Bulgaria, Kostinbrod.
Milbrath J. A., 1950. Latent ringspot virus of cherries
Science / Fund "Scientific Research" / - reduces growth of nursery trees. Plant Dis. Rep.
Contract DNTS/China 01/8/ 6.11.2014. 34:374-375.
Milbrath J. A., 1957. Effect of some sour cherry viruses
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Keulemans J., 2002. Effect of infection by viruses on Nemeth M., 1986. Virus, Mycoplasma and Rickettsia
vegetative and reproductive growth of sweet cherry Diseases of Fruit Trees. Akad. Kiado, Budapest,
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Aparicio F., Sánchez-Pina M. A., Sánchez-Navarro J. A., ringspot group, p. 104–132. In: Virus diseases and
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necrotic ringspot virus and prune dwarf virus in fruit 380-384.
trees. Tagungsberichte 97: 139-142. Posnette A. F., Cropley R., Swait A. J., 1968. The
Gilmer R. M., Nyland G., Moore J. D., 1976. Prune incidence of virus diseases in English sweet cherry
dwarf In: Virus diseases and noninfectious disorders orchards and their effect on yield. Ann. Appl. Biol.
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Agr. Res. Serv. Agr. Hdbk. 437, 104–132 Proebsting E. L., Ophardt D., Howell W. E., Mink G.I.,
Howell W. E., Mink G. I., 1984. Control of natural Patten, K.D., 1995. Variation in horticultural traits of
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20
MECHANICAL PROPERTIES OF THE FRUITS

OF SOME PERSPECTIVE SWEET CHERRY CULTIVARS
Simeon KRUMOV, Nicolai CHRISTOV
Institute of Agriculture - Kyustendil, 2500, Kyustendil, Bulgaria
Abstract
It was performed a research on the resistance of sweet cherry fruits to mechanical pressure and detachment force from
their fruits stem for the cultivars ‘Merchant’, ‘Pobeda Кrimska’, ‘Summit’, ‘Vanda’, ‘Stella’, ‘Sunburst’, ‘Rainier’,
‘Bing’, ‘Mizia’, ‘Kozerska’, ‘Vasinika’, ‘Alexton’ and control cultivar ʻVanʼ. The survey was conducted during
harvesting maturity of the fruits. These indicators characterize the transportability of the sweet cherry fruits and
suitability of the cultivars for mechanized harvesting. The results show that the fruits of all investigated cultivars had
from „good“ to „very good“ transportability, with the exception of ‘Pobeda Кrimska’ and ‘Sunburst’. The cultivar
‘Vasinika’, created at the Institute of Agriculture - Kyustendil, Bulgaria stands out with easily removable fruits, without
leakage of juice. This cultivar was the most suitable for mechanized harvesting from all investigated cultivars.
Key words: cultivar, mechanized harvesting, sweet cherry, transportability.
INTRODUCTION authors was found that the cultivar ‘Volsko’

had the weakest detachment force of the fruits
Along with the size and flavor, the density of from the stems-less than 250 g.
sweet cherry fruits is one of the main criteria It was most difficult to detach the fruit stems
determining the quality of fruits for fresh from fruits in cultivars ‘Badaconi’ and
consumption (Guyer et al., 1993; Bernalte et ‘Poznanska’ - from 531 to 740 g.
al., 1999; Vursavuş et al., 2006). The purpose of this study was to establish the
In some cultivars, the fruit flesh can be soft, in theoretical transportability and suitability for
others - tight in different degree (Georgiev et mechanized harvesting of some promising
al., 2007). sweet cherry cultivars.
According to Zhivondov et al. (2011), the later
ripening cultivars are characterized by good MATERIALS AND METHODS
density, crispness, tender skin, juiciness and
separability of the stone, while the early ones The research was conducted during the period
always have a soft or semi-thick texture. 2014-2016 at the experimental sweet cherry
In general the market preferences are targeted plantation of the Institute of Agriculture -
at sweet cherry cultivars having hard-fleshed Kyustendil, Bulgaria.
fruits, mainly because of their better transpor- The object of the study were cultivars
tability, storage and resistance to manipulation. ‘Merchant’, ‘Pobeda Кrimska’, ‘Summit’,
The strength of attachment of the fruits to the ‘Vanda’, ‘Stella’, ‘Sunburst’, ‘Rainier’, ‘Bing’,
fruit stems is directly related to the suitability ‘Mizia’, ‘Kozerska’, ‘Vasinika’ and ‘Alexton’.
of the cultivar for mechanized harvesting. The cultivar ʻVanʼ was used as control.
When detached, the fruit stem may tear the The trees were planted in the spring of 2002 at
fruit flesh or skin, and cause leakage of juice. distance of 6 x 5 m in the rows and formed in
The cultivars, which fruits are easily detached free growing crown.
from the stem, without tearing the skin and They are grafted on P. mahaleb rootstock IK-
flesh and no juice leakage, are suitable for M9, grown without irrigation. Each sweet
mechanized harvesting (Georgiev et al., 2007). cherry cultivar was represented by 5 trees.
In the investigation carried out by the same
21
The soil in the experimental plantation is pressures from 1812.9 to 2036.3 g had also the
highly leached, slightly sandy-clay cinnamon fruits of ‘Bing’, ‘Alexton’, ‘Kozerska’,
forest soil with a neutral reaction. ‘Rainier’ and ‘Vanda’. The cultivars ‘Vasinika’
The fruit transportability was experimentally (1675.7 g), ‘Stella’ (1495.0 g) and ‘Merchant’
determined by measuring the resistance of the (1461.0 g) are characterized by "good"
fruits to mechanical pressure until cracking of transportability.
the skin, and by measuring the detachment Relatively low resistance to pressure and "low"
force from their fruit stems. transportability, respectively, were found for
The measurements were carried out during fruits of the cultivars ‘Pobeda Кrimska’ (977.1
physiological maturity of the fruits, with a g) and ‘Sunburst’ (1277.7 g).
device made for this purpose, similar to the The strength of attachment of the fruit to the
apparatus АС 2 (Georgiev et al., 2007). fruit stem has a great practical importance.
The mechanical resistances most closely The cultivars, which fruits are easily to detach
imitate the transport pressures and the force of from the fruit stems, without tearing the skin
detachment of the fruits from their fruit stems and no leakage of juice, are suitable for
during harvesting. mechanized harvesting.
The indicators are measured in grams. After The data showed that during the study period
detachment of the fruit stems, it has been the smallest force of detachment of the fruits
reported whether there was observed any from their fruit stem was found in cultivar
tearing of the skin and leakage of juice. The ‘Pobeda Кrimska’ - from 220.5 to 240.7 g,
indicator is presented as a percentage of the followed by ‘Vasinika’ - from 335.0 g to 459.3
total number of fruits recorded. g. With "easy to medium hard" detachment of
There were investigated 3 replicates of 50 fruits the fruits were the cultivars ‘Merchant’ (from
per each cultivar. 409.6 g to 505.3 g), ‘Van’ (from 359.1 g to
The obtained experimental results were 578.0 g), ‘Sunburst’ (from 399.0 g to 595.3 g)
processed by the method of analysis of and ‘Stella’ (from 447.0 g to 571.3 g). For the
variance, using the LSD-test to prove statistical cultivars ‘Alexton’ (from 379.3 g to 610.0 g)
significance of the differences found between and ‘Rainier’ (from 450.2 g to 590.0 g),
the control and the variants. The evaluation was detachment of the fruits was "medium hard".
made at levels of significance P <0.05 , P <0.01 An average of 580.9 g to 622.3 g of force was
and P <0.001 (Maneva, 2007). needed for cultivars with "hard" detachment of
the fruits – ‘Vanda’, ‘Summit’ and ‘Mizia’.
RESULTS AND DISCUSSIONS "The most hard" was detachment of the fruits
in a cultivar ‘Kozerska’ from 689.5 to 871.7 g
The transportability of sweet cherry fruits is (Table. 1).
defined to a large extent by the biological The lower the value of this indicator, the more
characteristics of the cultivar. The rainfalls and suitable the cultivar for mechanized harvesting.
their amount during the ripening period, as well The statistical analysis of the results showed
as the applied agro-technology, also have an that proven differences were established
impact. between the cultivars ‘Mizia’ and ‘Summit’,
The results of our study show that, with the and the control in regard to resistance of the
exception of ‘Pobeda Кrimska’ and ‘Sunburst’, fruits to mechanical pressure.
the fruits of other sweet cherry cultivars had For the other cultivars the statistical proof is
good to very good transportability (Table 1). negative or insignificant.
As an average for the period of study with the The force of detachment of the fruits from their
highest endurance of mechanical pressure, stems was less than the control for cultivars
exceeding the value of the control ‘Van’ (2151 ‘Pobeda Кrimska’, ‘Vasinika’ and ‘Merchant’.
g), were the fruits of the cultivars ‘Mizia’ The difference between ‘Sunburst’ and the
(2278.3 g) and ‘Summit’ (2214.1 g). "Very control ‘Van’ was insignificant, and for other
good" transportable qualities, and resistance to cultivars - positively proven (Table 1).
22
Table 1. Mechanical properties of sweet cherry fruits, 2014-2016
Mechanical properties
The force of mechanical pressure The force of detachment of the fruits from
Cultivar of the fruits to crack the fruit stems
g g
2014 2015 2016 x* 2014 2015 2016 x*
1520.7 1427.9 1421.7 1461.0 505.3 409.6 358.0 424.3
‘Merchant’
−−− −−− −−− −−− − − n.s. −
1020.2 967.2 944.0 977.1 240.7 230.9 220.5 230.7
‘Pobeda Кrimska’
−−− −−− −−− −−− −−− −−− −−− −−−
2820.5 2031.8 1790.0 2214.1 784.2 581.1 396.0 586.7
‘Summit’
+++ −−− −−− + +++ +++ n.s. +++
1871.9 1801.9 1765.0 1812.9 753.1 560.4 429.2 580.9
‘Vanda’
−−− −−− −−− −−− +++ +++ ++ +++
1650.0 1469.9 1365.0 1495.0 571.3 465.0 447.0 494.4
‘Stella’
−−− −−− −−− −−− n.s. n.s. +++ +
1168.7 1249.5 1415.0 1277.7 595.3 420.0 399.0 471.4
‘Sunburst’
−−− −−− −−− −−− n.s. n.s. n.s. ns
1680.1 1640.0 1707.0 459.3 351.6 335.0 382.0
‘Vasinika’ 1675.7
−−− −−− −−− −−− −−− n.s. −−−
2148.2 1970.0 1833.0 1983.7 610.0 514.9 379,3 501.4
‘Alexton’
−− −−− n.s. −−− n.s. +++ n.s. +
2700.5 2249.5 1885.0 2278.3 750.2 569.8 547.0 622.3
‘Mizia’
+++ n.s. n.s. +++ +++ +++ +++ +++
1926.5 1990.0 1897.0 1937.8 590.0 450.2 504.0 514.7
‘Rainier’
−−− −−− n.s. −−− n.s. n.s. +++ ++
2116.0 2000.0 1800.0 1972.0 871.7 710.0 689.5
‘Kozerska’ 757.0
−−− −−− −− −−− +++ +++ +++
2222.6 1999.0 1887.3 2036.3 610.1 541.7 462.0 537.9
‘Bing’
− −−− n.s. −−− n.s. +++ +++ +++
‘Van’ (control) 2372.0 2195.5 1880.0 2149.1 578.0 445.0 359.1 460.7
F 133.6 275.3 222.9 384.1 53.9 106.2 63.4 124.2
SD 65.7 33.1 27.4 28.2 31.0 16.4 20.2 16.0
LSD (0.05) 135.4 68.2 56.4 58.1 63.9 33.9 41.6 32.9
* Average for the period 2014-2016
Figure 1 shows the average data for the cultivars with easily detached fruits, ‘Vasinika’
presence of skin tearing after detachment of the stood out with only 5.0% tearing of the skin
fruit stem from the fruit, as well as leakage of and without leakage of juice. As a result of the
juice. For the cultivars with "medium hard" and biological cultivar characteristics, the
"hard" detachment of the fruits, the value of the differences between the cultivars included in
indicators was low, and in five of them - the experimental work are significant.
‘Summit’, ʻMisiaʼ, ‘Kozerska’, ‘Bing’ and The variation of the indicators over the years is
ʻVanʼ control - zero. in a relatively narrow range. The absence of
An exception were ‘Sunburst’ and ‘Rainier’, big variations in the values means that the
where it was found that about 20% of the fruits studied cultivars are stable in terms of these
had torn skin and leakage of juice. From the important mechanical parameters.
23
%
Skin tearing Leakage of juice
55 60
50
30
40
20 22,5 30
19,4 1820
20
0 00
4,82 55 5
0
55
00 00 00 00
10
0
Figure 1. Condition of the fruits after detachment from the fruit stems
CONCLUSIONS The sweet cherry cultivar ‘Vasinika’, created at

the Institute of Agriculture - Kyustendil,
The transportability of sweet cherry fruits is Bulgaria stands out by "easily" detached fruits
defined to a large extent by the biological from stems, without leakage of juice. These
characteristics of the cultivar. The fruits of the qualities define it as the most suitable cultivar
investigated cultivars ‘Mizia’, ‘Summit’, for mechanized harvesting.
‘Bing’, ‘Alexton’, ‘Kozerska’, ‘Rainier’ and
‘Vanda’ had a "very good" transportability. ACKNOWLEDGEMENTS
With "good" transportable qualities were
characterized the fruits of ‘Vasinika’, ‘Stella’ This article is published with the financial
and ‘Merchant’, and with "low" - ‘Pobeda support of the Ministry of Education and
Кrimska’ and ‘Sunburst’. Science /Fund "Scientific Research"/ - Contract
A relatively "easy" picking of the fruits from DNTS/China 01/8/ 6.11.2014.
their stems was found for the cultivars ‘Pobeda
Кrimska’ and ‘Vasinika’. "Easy to medium REFERENCES
hard" for ‘Merchant’, ʻVanʼ, ‘Sunburst’ and
‘Stella’. "Medium hard" for ‘Alexton’ and Bernalte M., Hernández M., Vidal-Aragón M., Sabio E.,
ʻRaineerʼ. "Hard" detachment was found for 1999. Physical, chemical, flavor and sensory
characteristics of two sweet cherry varieties grown in
the cultivars ‘Vanda’, ‘Summit’ and ‘Mizia’, ‘Valle del Jerte’ (Spain). Journal of Food Quality, 22,
and "the most hard" - for the ‘Kozerska’. 403–416.
The absence of big variations in the values Georgiev V., Borovinova M., Koleva A., 2007. Sweet
means that the studied cultivars are stable cherry. Zemizdat, Sofia, 352 p.
regarding these important mechanical Guyer D., Sinha N., Tung-Sung Chang, Cash J., 1993.
Physicochemical and sensory characteristics of
parameters. selected Michigan sweet cherry (Prunus avium L.)
There are statistically proven differences cultivars. Journal of Food Quality, 16, 355–370.
between the cultivars ‘Mizia’ and ‘Summit’ Maneva S., 2007. Mathematical methods in plant
and the control in regard to resistance of the protection. PhD Thesis, Sofia, 201 p.
fruits to mechanical pressure. Vursavuş K., Kelebek H., Selli S., 2006. A study on
some chemical and physico-mechanic properties of
The force of detachment of the fruits from their three sweet cherry varieties (Prunus avium L.) in
stems is lower for the cultivars ‘Pobeda Turkey. Journal of Food Engineering, 74, 568-575.
Кrimska’, ‘Vasinika’ and ‘Merchant’ than in Zhivondov A., Malchev S., Tsareva I., 2011. Sensory
the control ʻVanʼ. profile and chemical components of fruits of sweet
cherry cultivars and elites. Plant science, 48, 24 – 30.
24

RESEARCH ON THE VARIETY INFLUENCE
ON VEGETABLE GROWTH AT APRICOT SPECIES
IN SANDY SOILS CONDITIONS IN SOUTHERN OLTENIA
Viorel ENACHE, Mihaela CROITORU, Milică DIMA
Research-Development Station for Plant Crop on Sands Dăbuleni, Dolj County, Romania
Abstract
The growth of shoots is a basic element in the relationship between growth and fructification, on which depends the
fruit trees equilibrium respectively the production provision for year and next years. Cuttings made during the spring
and green works on annual increases are a very important link in agrotechnics of intensive plantations, but at the same
time an element that loads the production cost. In conditions the sandy soils from Dăbuleni, fourteen apricot varieties
were studied, with different maturation periods, but also with differences in growth processes. Of the three experimental
years, the highest annual growth rates were recorded in year 2015 (158.21 cm average varieties) year in which both the
air temperature and the amount of rainfall were at normal values during the intensive growth of the shoots. The rhythm
of most intense growth was recorded in June and July, after which it diminished slightly in August and September as a
result of the ending vegetative increases of vegetative growth and the beginning of preparing the trees for the winter
dormancy. The year 2016, following the measurements made, showed smaller growth increases (124.21 cm), due to the
thermohydric stress conditions in the sandy soil area. In year 2017, on the background of optimum soil humidity the
growth of shoots was very intense as early as May. Due to the fact that in 2017 the trees presented fruit, the growth rate
decreased in June and July, and after the harvesting production, the vegetative growths again showed an intense
rhythm. The average value of varieties was 121.60 cm. The varieties that showed the highest annual increases in the
three years of study were 'Crystal' (130.2-190.2 cm) and 'Orizont' (132.8-199.2 cm).
Keys words: dynamic, annual growths, shoots, works in green.
INTRODUCTION of leaves and the foliar surface grow rapidly,

the leaves being of normal size (Baciu Adrian
The changes expected in the climate regime in A., 2005).
Romania fall within the global context, but In the juvenile period the interval that is
with particularities specific to the geographic sufficient for the tree not to suffer is between
region in which it is located. Compared with 10-20 branches on the tree. As trees grow, their
northwestern Europe, for example, where the number may increase, suggesting that 40% of
warmest heating is expected in winter, for the branches are vegetative (Mitrea, and
Romania, heating is expected to be more Tudosie, 2011).
pronounced during the summer (Sunley et al.,
2006). MATERIALS AND METHODS
In the intensive growth process, the synthesized
substances are consumed almost entirely, the The research was carried out at the, Research-
accumulations being poorly represented. In the Development Station for Plant Crop on Sands
phenophase of intense growth, trees have high Dăbuleni in the apricot species, with fourteen
nitrogen and water needs to form new tissues to varieties.
increase the volume and size of the crowns. The trees were planted at a distance of 4 m x 4
The intense growth phenophase of shoots is m, and the crown shape was flattened.
due to the production and spread of neoformed Determinations have been made on the growth
internodes, an activity that is growing more and rate of annual shoots from May to September
more accelerating. As a result of this intense during the period 2015-2017.
activity, the shoots grow in length, the number
25
RESULTS AND DISCUSSIONS 250 y = -0,111x2 + 4,456x
6x + 85,92
r=0,72** y = 0,275x2 - 0,271x + 123,2
28 June r=0,80**
Measurements of the growth rate of shoots in 2
200
y = -0,041x2
0 041x2 + 1,903x
1 903x + 55,59
555,59
5 59 26 July
Mean length of shoots per tree (cm)

apricot varieties were carried out in vegetation r=0,48
30 May
phases from May to the end of September. The 150
results obtained are presented in table 1. The

measurements included a group of fourteen 100
varieties with different ripening time and

highlighted very differentiated growth 50 y = 0,299x2 - 0,783x + 142,4
y = 0,214x2 + 0,841x + 132,7
r=0,77** r=0,77**
depending on the variety and climatic 28 August 28 September
conditions. 0
0 5 Soiul/variety 10
1 15
In the climatic conditions of year 2015, the
30.mai 28.iun
length of the shoots was of 77.8 cm at the 26.iul 28.aug
'Goldrich' variety, and 101.2 cm at the 'Cristal' 28.sep Poly. (30.mai)
Poly. (28.iun) Poly. (26.iul)
variety in May, and reached the end of the
growth at values ranged between 128.5 cm at Figure 1. The correlation between the average length of
the 'Goldrich' variety and 190.2 cm to the apricot shoots and the growth dynamics of apricot
'Cristal' variety (Table 1). varieties studied in the year 2015
Compared to the 'Olimp' control variety, the
annual growth rates were of 158.4 cm in Compared to 2015, in 2016, the measurements
September, higher values been determined to showed smaller growth increases between 50.7
the next varieties: 'Histria' (169.2 cm), 'Cristal' cm for 'Mamaia' variety and 76.5 cm for
(190.1 cm), 'Auraş' (161.4 cm), 'Ceres' (179 'Orizont' variety in May, and at the end of the
cm) and 'Orizont' (199.2 cm). increases, the length of the shoots was 104.6
The growth rate was very intense in June and cm in the 'Goldrich' variety and 146.8 cm in the
July, after that it slightly diminished in August 'Orizont' variety, but the rhythm of growth was
and September as a result of the interruption of much more intense (Table 2). The correlation
vegetative growth and the start of preparing factors determined were significant as early as
trees for winter dormans (Figure 1). May, from 0.73** to 0.82** in August, after
then fell to 0.77** (Figure 2).
Table 1. The dynamics growth of shoots at apricot
cultivars in 2015 Table 2. The dynamics growth of shoots at apricot
species in year 2016
Variety Mean length of shoots on tree to species
apricot (cm) Variety Mean length of shoots on tree to species
May June July August September apricot (cm)
May June July August September
'Fortuna' 65.2 96.9 118.7 131.5 142.3
'Fortuna 55.7 73.7 94.7 106.7 115.5
'Dacia' 58.3 97.5 125.5 142.7 145.7
'Dacia' 46.8 66.8 91.6 108.8 114.8
'Goldrich' 50.7 89.6 115.3 122.1 128.5
'Goldrich' 51.9 68.7 87.9 98.9 104.6
'Harcot' 66.7 110.7 145.5 161.3 165.8 'Harcot' 58.8 77.5 100.8 116.2 125.1
'Amiral' 62.3 98.7 129.9 132.4 134.6 'Amiral' 60.6 81.3 103.5 113.2 119.4
'Mamaia' 51.1 94.5 126.6 136.8 141.3 'Mamaia' 48.4 71.4 98.8 107.5 113.8
'Olimp' 'Olimp'
82.5 111.6 139.9 156.9 158.4 70.1 88.9 112.9 125.7 131.7
(Control) (Control)
'Augustin' 68.5 102.8 126.6 137.8 141.5 'Augustin' 61.4 81.3 104.1 117.2 123.1
'Histria' 54.7 116.4 145.5 168.7 169.2 'Histria' 49.9 72.9 91.9 107.3 112.1
'Cristal' 89.0 145.9 172.1 189.6 190.2 'Cristal' 79.8 102.7 121.3 132.4 139.2
'Auraş' 73.8 129.9 141.0 157.8 161.4 'Auraş' 67.8 92.8 113.7 124.8 131.5
'Euxin' 'Euxin' 65.8 87.8 110.4 122.6 126.5
70.8 118.7 140.0 153.5 157.6
'Ceres' 'Ceres' 72.6 95.4 118.6 129.9 134.8
71.3 133.6 162.1 176.7 179.3
'Orizont' 76.5 98.7 125.8 139.7 146.8
'Orizont' 71.4 110.7 188.8 196.9 199.2
26
The growth rate of shoots depends on the Table 3. The dynamics growth of shoots at apricot
variety, the climate conditions, the amount of species in year 2017
water in the soil and the supply of nutrients. Variety Mean length of shoots on tree to species
In the early years of planting the apricot has a apricot (cm)
rapid growth rate, forming shoots of 60-120 cm
'Fortuna 49.3 67.6 86.4 96.1 104.5
and numerous early shoots.
'Dacia' 50.1 73.1 97.4 109.9 118.1
160 y = 0,064x2 + 0,859x + 50,71 y = 0,039x2 + 1,656x + 67,57 'Goldrich' 45.6 64.6 82.5 104.6 110.5
r=0,73** r=0,81** 'Harcot' 105.6 113.4
140 56.2 79.2 95.5
30 mai 28 iunie
'Amiral' 58.5 79.8 104.1 113.9 120.6
120
'Mamaia' 52.3 70.2 90.3 101.5 108.8
100
'Olimp' 122.1 128.0
66.7 98.0 112.3
80 (Control)
'Augustin' 63.2 90.2 108.6 121.5 128.4
60 y = 0,132x2 + 0,104x + 113,7
r=0,77** 'Histria' 52.5 84.5 100.0 112.7 122.3
40 28 septembrie
y = 0,076x2 + 1,176x + 91,1
'Cristal' 71.4 93.4 108.8 120.4 130.2
y = 0,110x2
0 110x2 + 0,593x
0 593x + 105,4
105 4
20 r=0,82** r=0,82** 'Auraş' 72.4 97.4 110.3 123.6 128.8
26 iulie 28 august
0 'Euxin' 68.1 87.1 105.4 116.0 126.5
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 'Ceres' 123.0 129.3
74.3 101.3 114.1
Soiul/ Variety 'Orizont' 69.7 92.2 105.4 118.9 132.8
30.mai 28.iun 26.iul
28.aug 28.sep Poly. (30.mai)
Poly. (28.iun) Poly. (26.iul) Poly. (28.aug) 140 y = -0,023x2 + 2,350x + 44,81
1 y = -0,121x2 + 3,594x + 103,4
r=0,85** r=0,84**
Figure 2. The correlation between the average 120 30 mai 28 septembrie
tree length of apricot shoots and the growth dynamics
1
100
of the apricot varieties studied in the year 2016

80
In 2017, on the background of optimum soil
humidity the growth of shoots was very intense 60
y = -0,141x2 + 4,459x + 61,00
as early as May. 40 r=0,83**
Values were included between 45.6 cm in the 28 iunie y = -0,145x2 + 3,794x + 95,63
r=0,80**
'Goldrich' variety and 74.3 cm in 'Ceres' variety. 20 y = -0,155x2 + 4,059x + 82,29,2 28 august
Compared to the 'Olimp' control variety which 0
r=0,77**
26 iulie
in may the annual increases were 66.7 cm, the 0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16
'Cristal', 'Auraş', 'Euxin', 'Ceres' and 'Orizont' Soiul/ Variety
30.mai 28.iun 26.iul
varieties were recorded higher values between 28.aug 28.sep Poly. (30.mai)
68.1 cm in the 'Euxin' variety and 74.3 cm in Poly. (28.iun) Poly. (26.iul) Poly. (28.aug)
the 'Ceres' variety (Table 3).
Figure 3. The correlation between the average
Due to the fact that in 2017 the trees presented tree length and the growth dynamics of apricot
fruit, the growth rate decreased in June and varieties studied in the year 2017
July, and after harvesting production, the
vegetation growths again showed an intense
If we compare the three years of study, the
rhythm.
highest annual growth rates were recorded in
The correlation factors shown in (Figure 3)
2015, when both the air temperature and the
show the intensity of this process.
amount of rainfall were at normal values during
The highest annual growth rates were recorded
the intensive growth of the shoots.
in 2015, in which both the air temperature and
The highest values were determined for the
the amount of rainfall were at normal values
varieties: 'Cristal' (190.2 cm), 'Orizont' (199.2
during the intensive growth of the shoots.
cm) and 'Ceres' (179.3 cm) (Figure 4).
varieties: 'Cristal' (190.2 cm), 'Orizont' (199.2
cm) and 'Ceres' (179.3 cm).
27
250 The highest annual growth rates were recorded
199,2 in 2015, when both the air temperature and the
Mean length of shoots(cm)
200 190,2
169,2 179,3 amount of rainfall were sitated at normal values
158,4
150 during the intensive growth of the shoots.
100 varieties: 'Cristal' (190.2 cm), 'Orizont' (199.2
cm) and 'Ceres' (179.3 cm).
50
The growth rate of shoots depends on the
0 variety, climatic conditions, the amount of
water in the soil and the supply the tree with
nutrients.
Variety
Year 2015 Year 2016 REFERENCES
Year 2017
Baciu A., 2005. Pomicultură generală. Editura
Universitaria Craiova; 231-236.
Figure 4. Annual vegetative increases in apricot varieties Mitrea R., Tudosie R.E., 2011. The behavior of some
studied in the period 2015-2017 varieties of apple tree to the attack of pathogens
Venturia inaequalis (Cke.) Wint. and Podosphera
CONCLUSIONS leucotricha (Ell. Et Everh.) Salm, XVI (LII).
Sunley R.J., Atkinson C.J., Jones H.G., 2006. Chill unit
model sand recent changes in the occurrence of
In three years of experiments (2015-2017), winter chill and spring frost in the United Kingdom.
climatic conditions in 2015 have inprinted an Journal of Horticultural. Science & Biotechnology,
overlapping of phenophases of growth and 81: 949-958.
fructification earlier by about a week,
compared to 2017.
28
STUDIES REGARDING THE INFLUENCE OF PRE -HARVEST
AND POST - HARVEST TREATMENTS UPON THE QUALITY
OF SOME APPLE FRUIT VARIETIES
Adrian CHIRA1, Lenuța CHIRA1, Elena DELIAN1, Constanța ALEXE2

1
University of Agronomic Sciences and Veterinary Medicine of Bucharest,
59 Mărăşti Blvd, District 1, 011464, Bucharest, Romania
2
Research and Development Institute for Processing and Marketing of the Horticultural Products
Bucharest, No.1A, Intrarea Binelui Street, District 4, Bucharest, Romania, phone: 0770534388,
fax 0214600725, email: [email protected]

Abstract
Losses during storage of fruits are still considerable in some cases: about 20 - 30% of all produced harvested
worldwide is not consumed because of fungal or physiological disorders. In the present paper, we show the research
results of the pre-harvest treatments using the products: Rover - 0.2%, Sumilex - 0.1% and Topsin - 0.1%, as well as the
post-harvest treatments using: Rover - 0.2% and Sumilex - 0.1%. The treatments performed in the orchard before the
harvesting period have had a major effect to reduce the percent of rotten fruits in the storehouse. The reduction was
above 50% in the case of Rover - 0.2%, as compared with the untreated control. The studied apple varieties
(‘Jonathan’, ‘Generos’ and ‘Golden Delicious’) originated from the private farmers in Voinesti - Dambovita.
Key words: pre-harvest, post-harvest, rotten fruits.
INTRODUCTION vegetation period, but at the same time, imply a

special care of diminishing the pesticide
The biggest part of apples production is residuum on fruits (Bompeix G., 1985).
intended for storage, which allows the During storage period, apples can be attacked
commercialization for long periods after by a high number of fungus pathogens that
harvest (Hackbarth C. et al., 2017). cause their diseases. Infection can begin from
In the apple growth technology, the most the orchard or during transport and storage
important link is constituted by the period (Franchet J., 1991).
phytosanitary treatments performed in the
orchard, as well as during the storage period. MATERIALS AND METHODS
The losses due to the fungus impact during the
storage period are considerable, being up to The experience was carried out at Voinesti, in
20% from the total yield. the private orchards of some members of The
Improving the cultural practices and choosing Dambovita Fruit Growing Association.
the best varieties has an important contribution The purpose of this experience was that of
to yield increase and to the fruit quality. evaluating the apple fruits storage capacity and
Pre-harvest and post-harvest phytosanitary the maintenance of quality, following the
treatments represent an indispensable link for phytosanitary treatments applied in the orchard
apple culture. and after harvest, in the autumn of the year
Optimal postharvest treatments for fresh 2017. Fruits samples were also analysed, with a
produce seek to slow down physiological view to appreciate the physicochemical
processes of senescence and maturation, reduce characteristics, at the end of the storage period,
/ inhibit development of physiological disorders for ‘Jonathan’, ‘Golden Delicious’ and
and minimize the risk of microbial growth and ‘Generos’ varieties.
contamination (Mahajan et al., 2014). In our country the traditional variety is
Economical losses caused by parasite fungus ‘Jonathan’ (20%), but in the last decades new
justify the phytosanitary treatments during the and valuable varieties have been introduced,
29
such as ‘Generos’, ʻFlorinaʼ and so on. These Table 1. Pre-harvest treatments efficacy during storage
feature a genetic resistance to scab (Stanica Fl., period
Variety Variant Concentration Storage Rotten
2011). (%) period fruits
It is necessary to mention that in the orchard (days) (%)
the treatments were performed on 0.5 ha/ ‘Jonathan’ Control - 80 17.2
farmer, and after harvest the fruits were Rover 0.2 80 6.8
Sumilex 0.1 80 7.0
exposed to phytosanitary treatments, 100 kg Topsin 0.1 40 8.6
fruits on each variety. ‘Generos’ Control - 60 14.0
The fungicides used pre-harvest were Rover Rover 0.2 60 6.9
0.2%; Sumilex 0.1% and Topsin 0.1%. Sumilex 0.1 60 7.9
These were applied 20 days before harvesting Topsin 0.1 60 9.4
‘Golden Control - 95 19.0
and are recommended to prevent and control Delicious’ Rover 0.2 95 9.2
the major apple fruits storage diseases, Sumilex 0.1 95 10.4
produced by fungus: Penicillium sp.; Botrytis Topsin 0.1 95 11.3
cinerea and Gloeosporium album.
The fruits were stored in a unit with natural We can state that the treatments performed in
ventilation, with the following conditions: the orchard before harvest period have had a
temperature 14 - 150C and air relative humidity major effect on reducing the percent of rotten
70 - 75%. fruits in the storehouse. The reduction was
Spraying was performed as a post-harvest above 50% in the case of Rover 0.2%, as
treatment, using the products: Rover 0.2% and compared with the untreated control.
Topsin 0.1%. For the two experimental variants Also, it was observed that ‘Jonathan’ cv. the
both the fruits and the packaging were treated. principal pathogen was Penicillium sp. which
produce the moist rot, while for ‘Golden
RESULTS AND DISCUSSIONS Delicious’, the most important was the
lenticelary rot produced by the fungus
As far as the pre-harvest treatment is Gloeosporius album.
concerned, from the data presented in Table 1 it If we consider the storage period, that was 80
can be noticed that for all varieties, the best days for ‘Jonathan’, 60 days for ‘Generos’ and
results were obtained with the product Rover at 95 days for ‘Golden Delicious’, we can say that
0.2%. the last variety had a very good behaviour
The attack percent was 6.8% in the case of during storage, in relation with the major
‘Jonathan’; 6.9% for ‘Generos’ and 9.2% for pathogens.
‘Golden Delicious’, but after different storage As concerning the post-harvest treatment, as it
period, depending on the variety. can be observed in Table 2, these were more
From the tested products, the poorest results efficient than those performed during the
were obtained in the case of Topsin, the apple vegetation period, on the same product and at
fruits being attacked in a percent of 8.6% - the same concentration.
‘Jonathan’; 9.4% - ‘Generos’ and 11.3% - The Rover product in a concentration of 0.2%
‘Golden Delicious’. stood out again and it gave the best results.
The Sumilex product was more efficient than ‘Golden Delicious’ had a high percent of
the fungicide Topsin, but less efficient than the rotting fruits, as a consequence of the longest
Rover product. storage period.
30
Table 2. Pre-harvest treatments efficacy during storage period
Variety Variant Concentration Storage period Rotten fruits

(%) (days) (%)
‘Jonathan’ V1-Control - 80 13.2
V2-Rover (fruits) 0.2 80 5.9
V3 –Rover (wraps+fruits) 0.2 80 1.6
V4 –Sumilex (fruits) 0.1 80 6.6
V5 – Sumilex (wrap+fruits) 0.1 80 3.4
‘Generos’ V1-Control - 60 11.2
‘Golden Delicious’ V1-Control - 95 14.2
From the present data it can be noticed that the was a higher soluble carbohydrate content, a
pre-harvest and especially the post-harvest diminishing of fruits weight as a consequence
treatments - including wraps disinfecting - are of water losses, and a decrease of fruits
efficient to control pathogens during storage firmness because of pectin’s enzymatic
period. Finally, at the end of the storage period, breakdown.
physicochemical tests were run, with a view to In the case of fruit originated from the treated
characterise the fruit quality. Results are variants, the fruit storage capacity was better
presented in Table 3 and Table 4. and the qualitative characteristics were higher
It was emphasized that during the storage as compared to the untreated control.
period, the water contents decreased and there
Table 3. Fruits chemical analysis
Variety Water content Total dry Soluble dry Total acidity Acid ascorbic Minerals
(%) weight weight (%) (mg/100 g (%)
(%) (%) fw.)
‘Jonathan’ 81.40 18.60 15.60 0.27 3.96 0.25
‘Generos’ 80.50 19.50 14.90 0.43 4.76 0.27

‘Golden 78.60 21.40 15.10 0.18 4.25 0.23
Delicious’
Table 4. Fruits physical analysis

Variety Mean weight (g) Specific weight (g/cm3) Firmness (kgf/cm2)
‘Jonathan’ 120 0.744 3.9
‘Generos’ 135 0.780 3.7
‘Golden Delicious’ 138 0.760 3.6
31
CONCLUSIONS REFERENCES
The treatments performed in the orchard before Bompeix G., 1985. Traitment des pommes apres recolte
the harvesting period have had a major effect to contre les maladies de conservation. Larboriculture
fruitiere, 375, p. 37-40.
reduce the percent of rotten fruits in the Franchet J., 1991. Maladies de conservation des pommes
storehouse. The reduction was above 50% in et des poires. Phytoma, 432, p. 26-32.
the case of Rover 0.2%, as compared to the Hackbarth C. et al., 2017. Postharvest quality of Galaxy
untreated control. apples submitted to the treatment with 1 - MCP and
For all varieties, the lower rotting percentage different cooling times between harvest and cold
storage. Ciencia Rural, vol. 47, no. 8, Santa Maria.
has been registered for the variant were both Mahajan P.V. et al., 2014. Postharvest treatments of
the fruits and the wraps were treated, because fresh produce. Mathematical, physical and
these are an important source of pathogen engineering science, vol. 372.
infection. Stanica F., Braniste N., 2011. Ghid pentru pomicultori.
In the case of fruit originated from the treated CERES Publishing House, Bucharest.
variants, the fruit storage capacity was better
and the qualitative characteristics were higher
as compared with the untreated control.
32
SYSTEMS FOR SHAPING AND PRUNING OF PLUM TREES

USED IN BULGARIA
Denitsa SERBEZOVA
University of Forestry, 10 „St. Kliment Ohridski” Blvd, Sofia, Bulgaria
Abstract
The article presents an overview of the formation systems that have been imposed in the plum production in Bulgaria.
Crowns and pruning systems are directly related to the intended use of the fruit production, the planting density, the
degree of mechanization of the planting processes and harvesting. For fresh consumption (manual harvesting), at stem
height 50 -60 cm and planting distances are 5.0-5.5 m between the rows and 4.0 - 4.5 m in the rows, the trees are
formed as oblique palmette trees or fruit hedge (free palmette). For fruit processing such as dried, marmalade, mousse,
juice etc., mechanized harvesting requires stem height 90 - 100 cm at planting distances 6.5-7.0 m between the rows
and 4.5-5.0 m in the rows, trees are formed as modified central leader, semi-flat free crown and applying contour
pruning. For brandy and fruit spirit - the peculiarities of formation and pruning are described.
Key words: plum, pruning, forming systems.
INTRODUCTION The purpose of the currant study is to present the

most widely used systems for shaping and
Plum is an economically valuable fruit pruning plum trees in Bulgaria.
species with traditions of production in our
country. It occupies the second place after MATERIALS AND METHODS
sweet cherry, which is 6,705 ha, or 18 % of
areas under permanent crops in our country An in-depth analysis of the shaping and pruning
(MAF, 2017). systems used in Bulgaria was made.
Pruning and pruning systems in Bulgaria are Documents and literary, agrobiological and tech-
constantly evolving, due to the necessity to nological sources have been researched in order
increase yield and increase the quality of fruit to provide more complete information about the
production (Bozhkova 2006; Djouvinov and applied system formations in Bulgaria.
Vitanova, 2002; Djouvinov et al., 2012; Iliev A parallel is made between the systems used for
et al., 1977; Vitanova et al., 2010). shaping and pruning and are presented the most
The shaping and pruning of plum trees suitable for our country depending on the way of
depends on the variety and rootstock growth harvesting and the purpose of the obtained fruit
characteristics, the application of fruit pro- production.
duction (for fresh consumption or proce-
ssing), and the need to adapt the crown to RESULTS AND DISCUSSIONS
some of the mechanized agro-technical
activities (soil treatments, pruning, harvesting, Different types of crowns are known in practice,
etc.) (Djouvinov et al., 2012; Domozetov et which are divided into 3 main groups, depending
al., 2014; Petrov et al., 1979; Velkov, 1965; on the purpose of the resulting fruit production:
Vitanova et al., 2010). Hand fruit harvesting is used for fresh
The shaping and pruning of plum trees in the consumption. The trees are formed as oblique
production plantations must facilitate mecha- palmette trees or fruit hedge (free palmette) and
nized harvesting of fruit and contour pruning the planting distances are 5.0 to 5.5 m between
(Djouvinov and Vitanova, 2002; Petrov et al., the rows and 4.0 to 4.5 m in the rows.
1979; Sotirov et al., 2015). For processing fruits such as dried, marmalade,
mousse, juice etc., mechanized harvesting is
33
recommended, which requires a height of the Scaffold branches of the oblique palmette tree
stem 90-100 cm. Trees formed as modified formed only subscafold wood, evenly located
central leader, semi-flat free crown and sideways and outwards.
applying contour pruning. Planting distances It is used in varieties grafted on dwarf rootstocks.
are 6.5-7.0 m between the rows and 4.5-5.0 m This system requires a lot of manual work, strong
in the row. pruning and the cost of materials to build a
For brandy production and the production of supporting structure. It provides better crown
fruit spirit - the shaping and pruning of trees lightening, more effective plant protection,
for these production lines are related to the convenient for harvesting of high quality fruit.
simplification and reduction of the cutting First dormant pruning (pruning after planting).
operations in order to low quality require- Newly planted trees are cut to 80-90 cm above
ments of the resulting fruit production. the soil surface. Two opposing early twigs are
Plum trees used for the production of fresh selected to form the first floor, the strong ones
fruit in our country are formed as oblique cut to 15-20 cm and the weak to 4-5 buds.
palmette trees or fruit hedge (free palmette). In the first year, young trees should be grown so
These are semi-flat and flat crowns, which are that the leader and scaffolds grow longer than the
suitable for dessert varieties, the orientation of inter-floor distance.
the skeleton branches is in a plane in the Second dormant pruning. Suitable for scaffolds
direction of the row or with very little are opposite branches. It is desirable for
deviation from it. The total height of the tree scaffolds to select shoots aligned in length, thus
is 3.5-4 m and the stem 60-70 cm. saving extra measures to balance their growth.
For manual harvesting the planting distances Scaffolds are not abbreviated during the second
are 5.0-5.5 m between the rows and 4.0-4.5 m dormant pruning, as well as throughout the
in the row. Different systems of tree formation period. Head back leader to form the
formation are used. Formation and pruning of second floor. Cut at a height of 10-15 cm longer
oblique palmette plum trees. It is a flat than inter-floor distance. If the leader has not
artificial crown with a leader on which 2-3 reached that height by more than 10 cm, he does
floors are formed. Each floor consists of two not shorten.
scaffold branches located opposite that are Third dormant pruning. Two strong and aligned
directed in the direction of the row (Figure 1). lengths for scaffolds on the second set of
Thus, a continuous wall of scaffold and lateral branches are also choosen. Starters that make a
wood with a width of about 1.5-2 m is formed larger angle with the leader are preferred, and
in the row. their competitors are eliminated. The other lower
The distances between the scaffolds and the ones on the leader, with slight and moderate
number of branch set groups are determined growth, are not cut.
depending on the growth of the trees and the The leader shorten to 120-130 cm to form third
growing wood. In the case of highly growing set of scaffolds.
trees, three floors are formed at 110-120 cm The scaffolds on the first floor tilt if they are not
apart. inclined, the competitors are removed as well as
the stronger vertically growing branches.
Next pruning to complete the formation. In next
years trought the summer and dormant pruning
the third floor is shaped and the growth of the
already formed scaffolds is guided.
After the formation of the last floor, the leader is
not shorten. It is left without prunning for a few
years until the fruit grows, and growth is signi-
ficantly reduced, then reduced to 50-60 cm
above a moderately growing lateral branch with
fruit buds.
Until the trees come into full fruiting, the
pruning must be as weak as possible. Primary the
Figure 1. Oblique palmette plum trees
34
competitors of the leader ans scaffolds are re- For mechanized harvesting of plum fruits, the
moved. Some very strong and upright growing trees are formed as modified central leader,
branches, grown from the inside side of semi-flat free crown. The planting distances are
scaffolds, are also removed. Forming is easier 6.5-7.0 m between the rows, 4.5-5.0 m in the row
and faster when unnecessary shoots is done by and the height of the stem 90-100 cm.
summer pruning. Shortening of one-year-old Modified central leader is a naturally rounded
twigs is not recommended except for the length crown with a leader and 4-6 scaffolds, the first
of the leader to obtain branches for scaffolds. three forms the first floor and the next are
Pruning for shaping and growing of fruit located individually on the leader (Figure 2). It is
hedge (free palmette). This system is a free easy to create, the trees are vital and give very
palmette with a larger volume of the skeletal high yields.
part. It is used with moderate rootstocks.
First dormant pruning (pruning after plan-
ting). Trees cut down to 80 cm above the soil
surface. Weak twigs are shortened to 5-6 buds
and the strong ones to 15-20 cm, and those in
the stem area are cut to the base.
Second dormant pruning. Slight pruning is
done. Oppositely, disposed branches are se-
lected, with wide angle of deflection (above
45°) and grow in the direction of the scaffolds.
There is a head back the leader to form the
second floor. Cut at a height of 10-15 cm
greater than the specified inter-floor distance.
If the leader don’t exceeded this height by
more than 10 cm, it won’t be shortened.
Upright and upward-facing shoots are remo- Figure 2. Modified central leader
ved. For branching varieties ('Gabrovska', 'Pop First dormant pruning. The trees is cut to 100-
Hariton' etc.), the leader and scaffolds are not 130 cm. Premature twigs in the area of floor
cut, and those with weak branching ('Stanley' formation are shortened - weak to 4-5 buds, and
etc.) Require a moderate reduction in length. strong to 15-20 buds and lower located on the
Next pruning to complete the formation. stem part are removed to the base.
Using summer and dormant pruning in the Second dormant pruning. Appropriate twigs are
following years the remaining floors are for- selected for the main skeleton parts and the
med and the growth of the already formed branches of the main floor, observing the basic
scaffolds is guided. principles of pruning. The best-developed and
After the formation of the last floor, the leader centrally-occupied twig is chosen as leader.
is not abridged. It is left un pruned for a few From the sprouts grown under it, three scaffolds
years until the fruit grows, and growth is are selected which will form the floors which
significantly reduced, then reduced to 50-60 must be subordinate to the leader, well
cm above a moderately growing lateral branch developed and symmetrically located around the
with fruit buds. central axis, have a suitable angle of deflection
After the 5th-6th year, crown regulation is per- (45-55º) and, if possible be proportionate. The
formed, which can be done manually on con- remaining twigs are removed to the base.
tours or mechanized with RAC-6, in 2-3 years. Selected for scaffold branches are shortened in
Contour pruning can only be on the top, on length (60-70 cm) from the base, above the outer
both sides, with a triangular contour accom- or lateral bud. If their length is shorter, no
panied by a slight manual pruning even in the pruning is done. The leader cuts down in the area
middle years. on the second floor, if reached such a height.
The aim is to keep width of 3.0-3.5 m and a When there are no suitable branches for skeletal
height of 4.0-4.5 m of fruit hedge. ones or if the present ones are weak and the
leader is stronger, it is shortened by 15-20 cm
35
above the level of the scaffolds to dominate First dormant pruning. The trees are cut to 130-
over them. 140 cm from the soil surface. Premature twigs in
Third dormant pruning. One or two scaffolds the crown formation area are shortened to 15-20
on the leader on 80-90 cm are selected and the buds and removed in the stem area.
first subscaffolds on the scaffolds of the first
floor are determined. The leader is qoosen. Its
competitors are removed. Choose a suitable
branch for a scaffold, which should be well
developed, grow at a wide angle from the
leader and grow over one of the interstices of
the floor branches. The remaining strong
branches around them are removed, and the Figure 3. Schematic arrangement of the scaffolds and
weak, growing at right angles, are left over for subscaffold in semi-flat freely formed crown
growing wood.
The three scaffolds of the floor are chosen to Second - fifth dormant pruning. There is very
continue and their competitors are removed. little pruning, the strong and upright growing and
On each scaffold is selected for one branch branches in the intersection are removed.
for the first subscafold. For subscafolds, Trimming of the crown is done in two ways -
strong branches are used that grow sideways manually and mechanically.
and out of the scaffolds. Scaffolds are reduced Pruning fruiting trees. The balance between
to an outer bud of 70-80 cm above the growth and fruiting is good when the average
selected first skeletal branches so that they length of the trees of the fruit trees is not less
remain at one level. After headed back the than 20-25 cm, and the young - 50-60 cm.
leader continues to dominate over the It is achieved as 1-2 years of pruning and partial
scaffolds. rejuvenation.
The newly selected scaffold is shortened to Contour pruning is done to limit crowns to space
60-70 cm from the base to obtain the first between rows and height.
subscaffold. In the third dormant pruning, It can be done manually or mechanically with
only strong branches are removed, which are RAC-6 trimming. It can be carried out simul-
competitors of the leader, the selected taneously as a one-, two- or three-sided contour.
scaffolds and branches, and shorten their It can be held consecutively for 2-3 years.
lengths to obtain new scaffolds and subsca- Depending on the crown, the physiological state
folds. Pruning should be limited. of the trees in a given plantation determines the
Other pruning to complete the crown strength and the way of pruning.
formation. In the fourth and at the latest the For the successful application of industrial
fifth year, the formation of the crown is technologies and especially for the mechani-
finished, leaving the leader with 1-2 scaffolds, zation of harvesting the crowns of the trees should
20-40 cm apart, and on the scaffolds - another not exceed 4-4.5 m high and 4.5-5 m wide. After
1-2 scaffolds spaced apart 60-70 cm from the the contour pruning the trees get the shape of a
first, located on the other side of each cone by providing a free stripe for passage of the
scaffold. Annually, scaffold and one servicing technique (Figure 4).
subscaffold are left.
With the growing wood, the competitors of
scaffolds and the strong branches, growing in
the crown, the damaged and broken, are
removed. During the formation of the crown,
neither a severe reduction nor a strong dilu-
tion should be made in order not to cause
excessive growth and the fructification period
to be delayed.
Another mechanism used for mechanized har-
vesting is a semi-flat freely formed crown
Figure 4. Contour pruning scheme
(Figure 3).
36
Along with the contour pruning in the fruit- Operational Program "Science and education for
bearing plantations, different sanitary, intelligent growth", co-funded by the European
enlightening and detailed pruning can be Union through the European Structural and
carried out, taking into account varietal Investment Funds.
combinations and their growth and
reproduction. REFERENCES
Formation and pruning of trees for brandy
production and fruit spirit. Particularities Bozhkova V., 2006. Tree architecture for some plum
related to this particular trend are the more varieties. Collection of Reports of the First International
Symposium "Environmental Approaches to the
freely formed crowns, by reducing cutting Production of Safe Foods" 19-20 October 2006, 163-
operations and reduced fruit requirements. 168.
After the plum tree is fully harvested, the Djouvinov V., Vitanova I., 2002. Plum production in
pruning is minimized, but the fruit-bearing Bulgaria. Acta Horticulturae. 577, 25-31
wood has to be periodically rejuvenated. For Djouvinov V., Dinkova H., Bozhkova V., Minev I.,
Dragoyski K., Kutinkova H., Gercheva P., Nacheva L.,
4-5 years, a contour pruning is performed, Stefanova B., 2012. Plum. Biofruit Bg-Ltd., Plovdiv,
which is supplemented with a low manually 165-174.
pruning for shaping the crown. Domozetov D., Gandev S., Serbezova D., 2014. Shaping
and pruning of fruit tree and bushes. Biofruit Bg-Ltd.,
CONCLUSIONS Plovdiv, 55-57.
Iliev I., Vitanov M., Yoncheva M., Belyakov V., 1977.
Plum. Hristo Danov, Plovdiv, 139-151.
Systems for shaping and pruning plum trees Petrov A., Yoncheva M., Milanov B., Zhelev I., 1979.
in our country are differentiated according to Pruning of fruit plants. Hristo Danov, Plovdiv, 182-197
the purpose of the fruit production, which Sotirov D., Domozetov D., Borovinova M., Krumova A.,
determines the different height of the stem Krishkova I., Petrova V., 2015. Plum. Institute of
Agriculture - Kyustendil, Agricultural Academy - Sofia.
and the different degree of mechanization of 11-13.
the working processes for cultivation of the Velkov V., 1965. Pruning fruit trees. Fifth edition.
plum plantation. Zemzemat. Sofia. 140-149.
For manually harvesting the fruit, the trees are Vitanov, I., Marinova N., Ivanova D., Dimkova S.,
formed as flat and semi-flat crowns such as Kutinkova H., 2010. Plum Production in Bulgaria. Acta
Hortic. 874. 373-376.
oblique palmette trees or fruit hedge (free *** MAF, Department "Agrostatistics", Bulletin 326
palmette), as annual and high yields are "Fruit Production in Bulgaria in 2016", 2017.
obtained. https://goo.gl/vZ5GDa.
In mechanized harvesting, trees are formed as
modified central leader, semi-flat free crown
and applying contour pruning that is easy to
create, trees are vital and yields are very high.
For brandy production and fruit spirit - tree
pruning is weaker and crowns are formed
more freely to facilitate mechanized
cultivation and harvesting processes.
ACKNOWLEDGEMENTS
The article is published in collaboration with

Project 154 "Features in technology and
economic assessment of certain species and
varieties of perennial plantations in Bulgaria",
financed by the Science-Research Sector at
the University of Forestry and Project
BG05M2OP001-2.009-0034 "Support for the
development of scientific capacity at the
University of Forestry" financed by the
37
38
ASPECTS REGARDING THE QUALITY EVOLUTION OF SOME APRICOT

FRUIT VARIETIES DEPENDING ON THE STORAGE CONDITIONS
Lenuța CHIRA1, Adrian CHIRA1, Elena DELIAN,1 Ligia ION1, Constanța ALEXE2
1
2
Research and Development Institute for Processing and Marketing of the Horticultural
Products-Bucharest, No.1A, Intrarea Binelui Street, District 4, Bucharest, Romania,
phone: 0770534388, fax 0214600725, email: [email protected]
Abstract
For apricot fruits an important problem is to define the optimal picking time, the best storage variant and the variety
availability for storage as to avoid having different quality levels between on–tree and in-store maturity. In response to
this problem and some others that may arise, the research was performed in the orchard of the University of Agronomic
Sciences and Veterinary Medicine of Bucharest. Early results have revealed that fruits from 9 apricot cultivars
evaluated towards maturity in all the storage variants: NA-normal atmosphere (T=26°C), CA-cold atmosphere (T=2-
3°C, relative humidity 78 to 83%) and MA-modified atmosphere (T=2-3°C, relative humidity 85 to 90%) by wrapping
the storage case in plastic film. The MA storage variant may be positively considered since the total losses (both in
quantity and quality) were lower than in the NA and CA variants. The best response to temporary storage was found in
the late maturing varieties ʻFavoritʼ, ʻOlimpʼ and ʻExcelsiorʼ and in the semi-early maturing variety ʻDaciaʼ, the
storage period length being of 8 days in NA, 14 days in CA and 30 days in MA.
Key words: modified atmosphere, rottenness, storage condition.
INTRODUCTION MATERIALS AND METHODS
The present day opinion considers that the main A wide range of 9 varieties was analysed:
elements defining the quality of apricots, ʻDanubiuʼ, ʻOvidiusʼ, ʻFavoritʼ, ʻExcelsiorʼ,
mainly of those with double destination (fresh ʻMamaiaʼ, ʻDaciaʼ, ʻOrizontʼ, ʻOlimpʼ and
consumption and industrial processing) are the ʻHarcotʼ, which are well-known in Romania.
following: dry substance content of 18 - 20% The fruits were harvested from the orchard of
and over 20%, titratable acidity of 1 to 2%, the experimental area of the Faculty of
vitamin C of 20 - 25 mg/100 g, proteins 2 to Horticulture, in Bucharest.
2.5%, pectines 0.7 to 1%, average fruit weight We recorded the fruit bearing phases, the
of 65 to 80 g and over 80 g, appealing physical and chemical features of the fruits,
appearance, authentic apricot fragrance defined when harvested as in-completely and
(Alejandro Perez-Pastor et al., 2007). fully maturated and at in-store maturity,
The question that arises here is about the weight, weight losses and quality drops
correlation that may be carried forth between throughout storage, with special reference to
the levels obtained by the components of fruit the attack of Monilinia sp.
quality of valuable cultivar at wax ripeness and The fruits were stored in the facilities of the
at on - tree maturity and at in-store maturity laboratory existing at the Horticultural Faculty,
stages (Hitka, 2011). being conducted in three variants, and each
The temporary storage of the fruit depends on variant was repeated three times.
temperature and relative air moisture. The NA – normal atmosphere, namely storage in
damage caused by very high or low shed located in the orchard in a shady place and
temperatures results in tissue decomposition at an ambient temperature of 26°C.
(Bălan et al., 2008).
39
CA – cold atmosphere, 2 to 3°C temperature matter content, overall sugar and ascorbic acid
and 78 to 83% relative humidity. content (Table 1 and Table 2).
MA – modified atmosphere achieved by Higher gains were recorded in the ʻDanubiuʼ
wrapping the storage case in plastic film at a variety: where the increase from wax ripening
temperature of 2 to 3°C. to full maturity was of 5% in soluble dry matter
and the ʻFavoritʼ variety recording 10.1%
RESULTS AND DISCUSSIONS increase of soluble dry matter at the beginning
of ripening and a fruit size of 59 g with an
Physicochemical features of the fruits increase of up to 16% of soluble dry matter and
Harvesting at wax ripening and at full maturity a growth of up to 68.5g of the fruit; ʻExcelsiorʼ
with a content 9.4% soluble dry matter at wax
By comparing the levels of some essential ripening and 14.6% at full maturity and
components of fruit quality we noticed an ʻDaciaʼ, where the levels increased by 5% for
evolutive change in fruit size, in soluble dry soluble dry matter and by 15 g fruit weight.
Table 1. The main physical and chemical characteristics of the fruits of some apricot varieties
as determined at picking in the wax-ripe phase
Variety Average Total sugar Soluble dry Titratable Total Ascorbic acid
weight (g) (%) matter (%) acidity (%) sugar/titratable (mg/100 g)
acidity ratio
ʻDanubiuʼ 55 7.2 9.0 2.21 3.26 7.04
ʻOvidiusʼ 50 9.44 11.8 2.30 4.10 5.28
ʻFavoritʼ 59 8.08 10.1 2.04 3.96 5.28
ʻExcelsiorʼ 68 7.52 9.4 1.95 3.86 5.28
ʻMamaiaʼ 65 7.84 9.8 1.48 5.30 6.16
ʻDaciaʼ 52 9.12 11.4 2.05 4.45 5.28
ʻOrizontʼ 65 10.4 13.0 1.90 5.46 5.28
ʻOlimpʼ 73 8.8 11.0 1.61 5.46 4.4
ʻHarcotʼ 74 9.44 11.8 1.82 5.19 4.52
Table 2. The levels of some fruit quality in apricot varieties at full maturity
Variety Average Soluble dry Total sugar Titratable Total Ascorbic acid
weight (g) matter (%) (%) acidity (%) sugar/titratable (mg/100 g)
acidity ratio
ʻDanubiuʼ 57.5 14.0 12.3 2.02 6.1 16.19
ʻOvidiusʼ 53.7 13.0 10.0 2.20 4.5 11.50
ʻFavoritʼ 68.5 16.0 15.3 1.91 8.0 12.60
ʻExcelsior 70.6 14.6 13.0 1.67 7.8 11.62
ʻMamaiaʼ 67.8 13.0 10.3 1.22 8.4 11.52
ʻDaciaʼ 67.3 16.0 13.4 1.50 8.9 14.40
ʻOrizontʼ 68.5 15.0 12.3 1.32 9.3 12.02
ʻOlimpʼ 65.7 14.0 12.6 2.12 5.9 15.40
ʻHarcotʼ 67.2 15.0 12.8 1.40 6.4 13.80
Evolution of the main physicochemical features In the cold atmosphere variant at 2 to 3°C
during in-store fruit maturity temperature and 78 to 83% relative humidity
the increase in sugar content was lower than in
The overall sugar content recorded by us the normal atmosphere, still higher than in the
increased irrespective of the storage variant or modified atmosphere (Table 3). Difference
cultivar, except for Favorit, where a slight between varieties also occurred as far as the
decrease under modified atmosphere conditions content of sugar during storage was concerned.
was recorded.
40
Table 3. The main chemical characteristics of the fruits of some apricot varieties as determined after storage period
Variety Storage Soluble dry Total sugar Titratable Total Ascorbic acid
conditions matter (%) (%) acidity (%) sugar/titratable (mg/100 g)
acidity ratio
ʻDanubiuʼ NA 10.0 8.0 1.6 5.0 6.06
CA 9.5 7.6 1.7 4.5 6.26
MA 10.0 8.0 1.9 4.2 6.45
ʻOvidiusʼ NA 12.5 10.0 1.6 6.2 4.25
CA 12.0 9.6 1.8 5.3 4.65
MA 12.0 9.6 2.0 4.8 4.80
ʻFavoritʼ NA 14.4 11.5 1.6 7.2 4.05
CA 14.2 11.4 1.8 6.3 4.35
MA 10.0 8.0 1.9 4.2 4.57
ʻExcelsiorʼ NA 10.5 8.4 1.5 5.6 4.15
CA 10.0 8.0 1.6 5.0 4.45
MA 9.5 7.6 1.7 4.5 4.70
ʻMamaiaʼ NA 12.0 9.6 1.3 7.4 5.16
CA 12.5 10.0 1.4 7.1 5.46
MA 13.0 10.4 1.5 7.4 5.70
ʻDaciaʼ NA 15.2 12.2 1.5 8.1 4.00
CA 15.0 12.0 1.6 7.5 4.20
MA 1.,5 10.0 1.9 5.3 4.52
ʻOrizontʼ NA 15.9 12.7 1.4 9.1 3.92
CA 15.5 12.4 1.6 7.7 4.11
MA 14.0 11.2 1.8 6.2 4.40
ʻOlimpʼ NA 8 10.2 6.7 16.9 3.80
CA 14 14.3 6.5 20.8 3.91
MA 30 4.1 2.3 6.4 4.00
ʻHarcotʼ NA 8 7.6 6.9 14.5 4.92
CA 14 15.8 6.7 22.5 5.02
MA 30 4.2 2.4 6.6 5.15
NA = Normal atmosphere; CA = Cold atmosphere; MA = Modified atmosphere.
In a normal atmosphere the highest rises were (ʻMamaiaʼ) to 1.8% (ʻOvidiusʼ and ʻFavoritʼ);
found in the ʻFavoritʼ variety (by 3.4%), while in the modified atmosphere the limits
followed by ʻDaciaʼ (3.08%), ʻHarcotʼ (2.76%) were from 1.5% (ʻMamaiaʼ) to 2% (ʻOvidiusʼ).
and ʻOrizontʼ (2.3%). In cold atmosphere the Storing in modified atmosphere caused a higher
ʻFavoritʼ variety recorded rises of 3.32%, level of organic acids following the inhibitive
ʻDaciaʼ 2.28%, ʻHarcotʼ 2.56% and ʻOrizontʼ effect of the CO2 content increase into the
2%. In modified atmosphere, the varieties dehydrogenase activity.
ʻDaciaʼ, ʻOrizontʼ and ʻOlimpʼ showed A decrease in the ascorbic acid content was
increases of only 0.8%. A different behaviour between the beginning of the ripening and the
was obvious with the ʻMamaiaʼ variety, which post maturity stage in all the investigated
recorded a 2.56% increase and the ʻFavoritʼ, varieties, with different intensities in each of
where the sugar content drop correlated with the three storage variant.
the flesh browning was evident. The normal atmosphere variant recorded an
Titratable acidity also recorded different ascorbic acid content as low as 3.8 mg/100 g
evolutions, depending on the variety and the (ʻOlimpʼ) and 6.06 mg/100 g (ʻDanubiuʼ).
storage method. Its lowest value was recorded Higher levels were recorded in the cold
in normal atmosphere, caused by the speeding atmosphere variant, of 3.91 mg/100 g (ʻOlimpʼ)
up of oxidation at 26°C. and 6.26 mg/100g (ʻDanubiuʼ). The highest
While at harvesting at wax ripening the levels were recorded in the modified
titratable acidity limits ranged from 1.48% atmosphere variant (ʻOlimpʼ variety featuring 4
(ʻMamaiaʼ) to 2.3% (ʻOvidiusʼ), in the normal mg/100 g) and ʻDanubiuʼ to 6.46 mg/100 g
atmosphere the limits ranged from 1.3% ascorbic acid. The value of the overall
(ʻMamaiaʼ) and 1.6% (ʻOvidiusʼ). In the cold sugar/titratable acidity ratio that is a milestone
atmosphere the limits were from 1.4% for assessing the gustative quality of
41
horticultural products, encountered changes variants, as it is revealed in Table 4. Thus, the
throughout the post-harvest period, following optimal storage duration in normal atmosphere
the unequal rate of these two components. was of 8 days, 14 days in cold atmosphere and
When harvesting at wax ripeness, this ratio 30 days in modified atmosphere. Overall losses
ranged from 3.26 (ʻDanubiuʼ) to 5.46 (quantitative and qualitative) in fruits stored in
(ʻOrizontʼ and ʻOlimpʼ) going up to 5 (normal normal atmosphere after 8 days were high
atmosphere variant), 4.5 (cold atmosphere) and because of the high temperature and low rela-
4.2 (modified atmosphere) for ʻDanubiuʼ and to tive humidity, ranged from 14.5% in ʻHarcotʼ
9.1 (normal atmosphere), 7.7 (cold atmosphere) variety and 26.7% in ʻOvidiusʼ variety.
and 6.2 (modified atmosphere) for ʻOrizontʼ In the case of cold atmosphere, high overall
while for ʻOlimpʼ the results were 7.5 (normal losses were also encountered, ranging from
atmosphere), 7.8 (cold atmosphere) and 6.4 13.2% in ʻOrizontʼ and 27.9% in ʼDanubiuʼ,
(modified atmosphere). A higher value of the that is close to previous values (normal
ratio under normal atmosphere conditions and a atmosphere), attained however after a 14 days’
difference between the ratio in cold atmosphere laps of time. The overall losses after 30 days of
and that in modified atmosphere were storage under modified atmosphere were
highlighted because modified atmosphere clearly lower than those in the two preceding
slows down the metabolism of organic acids variants. Thus, weight losses ranged from 1.5%
and of glucides, which results in failing to (Excelsior) to 5.3% (ʻDanubiuʼ). While the
attain the characteristic gustative features. qualitative ones (Monilinia sp. attack or inner
browning) ranged from 2.7% (ʻOrizontʼ and
Capacity of temporary storage of apricot 14.5% ʻFavoritʼ). The ʻFavoritʼ variety, as
varieties already mentioned, proves improved behaviour
under these conditions, since fruits do not
The duration of apricot fruits storage picked at mature and the flesh becomes brown.
the wax ripening phase differed in the three
Table 4. Behaviour of the apricot fruits varieties harvested at wax ripeness
Variety Storage conditions Storage period (days) Weight losses (%) Quality losses (%) Total losses (%)
ʻDanubiuʼ NA 8 9.8 10.5 20.3
CA 14 21.4 6.5 27.9
MA 30 5.3 3.4 8.7
ʻOvidiusʼ NA 8 14.4 12.3 26.7
CA 14 20.2 6.8 27.0
MA 30 5.2 3.1 8.3
ʻFavoritʼ NA 8 12.1 13.5 25.6
CA 14 20.1 7.3 27.4
MA 30 2.6 14.5 17.1
ʻExcelsiorʼ NA 8 14.6 10.7 25.3
CA 14 11.1 5.8 16.9
MA 30 1.5 2.7 4.2
ʻMamaiaʼ NA 8 18.0 6.2 24.2
CA 14 1.1 5.5 16.6
MA 30 1.6 3.4 5.0
ʻDaciaʼ NA 8 13.4 7.5 2.,9
CA 14 11.9 5.9 17.8
MA 30 4.2 2.3 6.5
ʻOrizontʼ NA 8 9.0 6.3 15.3
CA 14 7.5 5.7 13.2
MA 30 3.1 1.7 4.8
ʻOlimpʼ NA 8 10.2 6.7 16.9
CA 14 14.3 6.5 20.8
MA 30 4.1 2.3 6.4
ʻHarcotʼ NA 8 7.6 6.9 14.5
CA 14 1.8 6.7 22.5
MA 30 4.2 2.4 6.6
42
CONCLUSIONS value throughout the storage period, if
compared to the normal atmosphere and cold
In order to obtain highly qualitative fruits as far atmosphere variants.
as the organoleptic aspect is concerned, after
their temporarily storage under various REFERENCES
conditions, fruits picked at wax ripeness period
must feature values of the overall PérezǦPastor A., RuizǦSánchez M.C., Martínez J.A.,
sugar/titratable acidity ratio of 3.26 to 5.46. Nortes P.A., Artés F., Domingo R., 2007. Effect of
deficit irrigation on apricot fruit quality at harvest and
The optimal storage period for apricot fruits during storage. Journal of the Science of Food and
picked at wax ripeness stage was of 8 days in Agriculture, 87, 2409 – 2415.
normal atmosphere, 14 days in cold atmosphere Bălan V., Stanică F., Chira L., Asănică A., Oprea M.,
and 30 days in modified atmosphere. Topor E., Hoza D., Marin D., Corneanu M., Tudor
The capacity of apricot fruits which were V., Chira A., Nistor E., Chiriceanu C., Ştefan S.,
2008. Caisul si caisele. CERES Publishing House,
temporarily stored varied within close limits, Bucharest, 564.
the best results were recorded in ʻOrizontʼ, Hitka G., 2011. Development of the controlled
ʻOlimpʼ, ʻDaciaʼ and ʻHarcotʼ varieties. atmosphere storage technology of apricot. Doctoral
The modified atmosphere obtained inside the theses, Faculty of Food Science Corvinus, Budapest,
storage package wrapped in semipermeable 75 – 78.
plastic film keeps the overall losses at a low
43
44
ECONOMIC EFFECT OF SUSTAINABLE APPLE PRODUCTION
Iliyana KRISHKOVA1, Denitsa SERBEZOVA2

1
Institute of Agriculture - Kyustendil, 1 „Sofiysko shose” Street, Kyustendil, Bulgaria
2
University of Forestry, 10 „St. Kliment Ohridski” Blvd, Sofia, Bulgaria
Abstract
The experiment was carried out during the period 2014-2016 at the Institute of Agriculture - Kyustendil using ʻFlorinaʼ
and ʻFreedomʼ cvs. grafted on seedling rootstocks of Winter Gold Pearmain and wild apple cultivated in a high density
plantation with trees of the same cultivars on clonal rootstocks MM 106 (in the row) and M 9 and Marga Hndzor (MH)
(between the rows). The soil is chromic luvisols. In order to study the influence of the intercropping on the growth and
production of the trees, there are 3 experiments – cultivar-rootstock trail without intercrop, cultivar-rootstock trail with
intercrop in rows and inter rows spacing. The production costs required for the cultivation of one hectar apple tree
orchard of the studied combinations range from 1900 euro/ha from Freedom of Winter Gold Pearmain to 282 euro /ha
at Freedom on Winter Gold Pearmain with interplants on MM 106 and M9. Cost differences are the result of the
number of trees per hectar, the average yields and the associated costs of collecting the additional fruit production. The
average yields and the resulting gross output have a significant impact on the net profit. In ʻFlorinaʼ variety, the most
effective combination of wild apple rootstock with in row intercrop of MM 106, and in the inter row of MH. For
Freedom, better results have been obtained using a Winter Gold Pearmain with intercrop on M9 rootstock, indicating
that for the different varieties it is necessary to select suitable rootstocks.
Key words: apple, rootstock, yield, quality, economic analysis.
INTRODUCTION found that the density of the trees and the

variety influence total
Numerous studies in apple plantations have yield. So, for ‘Auriu de Bistrita’ cv., the yield
shown that optimal planting density is the at density 2,500 exceeds control with 53% and
dominant factor for yields per unit area. A for 5,000 with 112% and for ‘Florina’ cv. with
decisive role in increasing the density of apple 69.3 and 135.5%, respectively (Platon et al.,
plantation has two main factors - rootstock and 2014).
variety. The use of dwarf rootstock allows
Trees of ʻJohnagoldʼ variety on MM 106 of the intercropping of the area, early fruiting of trees,
following planting distances were tested: 4 x 1, sustainable productivity, good fruit quality and
4 x 1.5, 4 x 2, 4 x 2.5 and 4 x 3 m, with the high economic efficiency (Dyankov, 1995;
largest cross-sectional area of the stem, the Domozetov and Radomirska, 2009).
largest crown volume and the highest yield In the analysis of the economic performance of
being obtained at a distance of 4 × 3 m. The apple plantation grown at three planting
results show that planting distances have a densities, an optimal density of 2,500 trees/ha
negligible effect on the quality of the fruit but was established (Fett and Waquil, 2001).
are important for the coloring (Kiprijanovski et The aim of the this study is to make a com-
al., 2009). In an experiment in Ireland with an parative analysis of variety-rootstock combina-
M9 rootstock at a planting density of 672, 961 tions grown as an intercrop apple plantation.
and 1,492 trees/ha, and M27 at a density of
1,279, 1,492 and 1,957, it was found that M9 MATERIALS AND METHODS
had the highest yield at the highest density
(Mac an Saoir et al., 2014). In Romania, an To investigate the effect of the use of intercrops
experiment with the apple ʻFlorinaʼ and ʻAuriu in apple plantations with different rootstocks,
de Bistriţaʼ cvs., grafted on M9 and using a three planting experiments were established in
different planting density (1,666,500 and 5,000 the spring of 1998, with ʻFlorinaʼ and
trees/hectare) formed in the ʻVʼ system, it was ʻFreedomʼ trees grafted on the seedlings of
45
Winter Gold Pearmain (WGP) and wild apple The influence of the kind of the rootstock is
(WA) and on the vegetative rootstocks - clearly expressed, taking into account the
MM106, M9 and Marga Hondzor (MH): average values of the stem diameter of all
1. Variety-rootstock trail without intercrop variants. Trees of both varieties on seed
- the planting distance between rows is 8 m and rootstocks have a larger stem diameter than
in the row 7 m or 179 trees/ha; those on MM 106, M9 and MH. Difference was
2. Variety-rootstock trail with intercrop in found for both types of seed rootstocks in
the row - on the seed rootstock (main trees) the variants with and without intercrops. The
planting distances are 8 x 7 m or 179 trees/ha. differences with Winter Gold Pearmain are
Trees - intercrop in the row on vegetative statistically proven. The thicker tree stem is
rootstock MM106 are planted (over one of the explained by the growth force of the seed
main trees) at 7 m (8 m x 7 m or 179 trees/ha). rootstock, which induces a stronger thickening
A total of 358 trees/ha; of the tree stem of the used grafts. When
3. Variety-rootstock trail with intercrop in comparing the influence of the variety on the
rows and between rows - on seed rootstock thickening of the stem, it can be seen that the
(main trees) planting distances are 8 x 7 m or ʻFlorinaʼ trees have a thicker stems than those
179 trees/ha. Trees - intercrops in rows on of ʻFreedomʼ and the different types of
vegetative rootstock MM 106 are planted (over rootstocks, except for MM106 and MH.
one of the main trees) at 7 m distance (8 m x 7 The crown height data for both varieties shows
m or 179 trees/ha) and between rows on that trees on seed rootstock tended to have a
vegetative rootstock M9 and MH - 2 m (8 x 2 higher crown height than those of the MM 106,
m) or 625 trees/ha. Total of 980 trees/ha. M9 and MH clone rootstocks. Trees of
During the fruiting period, the main economic ʻFlorinaʼ cv. on different types of rootstocks
indicators are calculated annually: gross output, have a larger crown volume than those of
euro/ha; production costs, euro/ha; net income, ʻFreedomʼ (Table 2).
euro/ha; rate of profitability, %. The necessary Average, for the period 2014-2016, tree yield
means of production are established on the for non-intercrop variants is highest in ʻFlorinaʼ
actual costs incurred on the basis of the norms on a WGP rootstock that amounts to 122.77
and tariffs for manual and mechanized works in kg/tree, followed by trees on a wild apple
the Institute of Agriculture - Kyustendil and the rootstock - 89.4 kg/tree (Table 3).
prices of the raw materials and supplies. The
Table 3. Yield, mean weight and fruit quality
valuation of the output was made at the actual
realization prices during each year. Yield Average
Quality, %
Planting
Variety Rootstock per fruit
combination
tree, kg weight, g Extra I II
RESULTS AND DISCUSSIONS
122.77 115.67 54.95 30.00 15.05
ʻFlorinaʼ WGP
The values for the average stem diameter are Main trees
without WA 89.40 116.67 63.17 24.55 12.28
constantly increasing for trees of all variants intercrop WGP 61.80 116.67 69.53 15.28 15.19
ʻFreedomʼ
for both varieties. In 2014, the largest diameter WA 64.23 115.67 66.75 18.00 15.25
of the stem was the ʻFlorinaʼ tree on the WGP 44.80 124.33 72.50 18.59 8.90
WGP
without intercrops and pruning - 21.60 cm and ʻFlorinaʻ
Main trees 68.73 126.33 82.97 12.24 4.79
the smallest with a intercrop of MH - 11.40 cm with
WA
(Table 1). In the ʻFreedomʼ cv.with the highest intercrop
WGP
60.60 91.33 47.84 24.19 27.97
thickness are the trees grafted on WGP, but ʻFreedomʼ
49.50 118.67 76.86 12.66 10.48
with a intercrop - 17.5 cm, and the smallest at WA
MM 106 42.97 113.83 63.51 24.36 12.13
M9 - 11 cm. Trends in ʻFlorinaʼ are preserved ʻFlorinaʼ M9 20.17 115.00 54.67 29.83 15.50
in the years 2015, 2016 and 2017, with the Trees - MH 18.40 124.00 73.12 19.76 7.12
growth of the stem being greatest in the trees intercrops MM 106 50.55 80.67 33.02 23.42 43.56
grafted on wild apple with a intercrop - 1.36 cm ʻFreedom M9 40,23 92,33 53,58 19,22 27,20
MH 31,97 70,00 21,27 24,54 54,19
and smallest on those with intercrop on M9 -
0.54 cm. In ʻFreedomʼ cv., this trend is
preserved except for trees grafted on MM106.
46
Table 1. Diameter of stem, cm
ʻFlorinaʼ 2014 2015 2016 2017 Growth, cm

WA without intercrop (st) 19.42 19.94 20.17 20.55 1.13
WGP without intercrop 21.33 ++ 21.86 ++ 22.13 ++ 22.47 ++ 1.14
WGP without pruning 21.60 ++ 21.87 ++ 22.29 ++ 22.62 ++ 1.02
WA with intercrop 18.86 19,24 19.68 20.22 1.36
WGP with intercrop 18,19 + 18.49 + 19.01 19.09 + 0.90
ММ 106 – in row intercrop 14.27 14.61 14.85 15.05 0.78
М 9 – between row intercrop 12.30 12.58 12.66 12.84 0.54
MH – between row intercrop 11.40 11.60 11.76 11.98 0.58
Sd 0.5004 0.5565 0.5853 0.5567
Gd 0.5 1.09 1.213 1.275 1.213
f 17.4407 15.1577 12.7847 14.8716
47
ʻFreedomʼ 2014 2015 2016 2017 Growth, cm
WA without intercrop (st) 16.64 16.76 17.48 18.15 1.51
WGP without intercrop 16.72 17.24 17.96 18.19 1.47
WA with intercrop 17.16 17.36 18.31 18.75 1.59
WGP with intercrop 17.52 17.88 18.35 18.75 1.23
ММ 106 – in row intercrop 15.16 14.77 15.05 15.13 0.72
М 9 – between row intercrop 11.03 11.22 11.42 11.90 0.87
MH – between row intercrop 11.80 11.98 12.30 12.74 0.94
Sd 0.8850 0.7926 0.9327 0.9597
Gd 0.5 2 1.791 2.107 2.168
f 0.4309 0.6703 0.3778 0.2420
Table 2. Size and volume of the crown
2014 2015 2016 2017
h-height, d-width, V- h - height, d-width, V-volume, h-height, d-width, V-volume, h-height, d-width, V-volume,
Variety m length, m volume, m3 m length, m m3 m length, m m3 m length, m m3
ʻFlorinaʼ
WA without intercrop (st) 3.65 3.86 14.23 3.31 4.10 14.56 3.75 3.79 14.09 3.18 4.04 13.58
WGP without intercrop 3.92 4.12 17.41 ++ 4.07 4.40 20.62 ++ 3.92 4.23 18.35 + 3.94 4.75 23.26 +++
WGP without pruning 3.82 4.13 17.05 + 3.75 4.36 18.65 + 4.22 4.47 22.06 ++ 3.56 4.40 18.03 ++
WA with intercrop 3.62 3.22 9.82 ++ 3.50 3.25 9.67 + 3.88 3.34 11.32 3.20 3.31 9.17 ++
WGP with intercrop 3.38 3.13 8.66 +++ 2.75 3.06 6.74 +++ 3.52 3.10 8.85 + 3.09 3.26 8.59 ++
ММ 106 – in row intercrop 2.95 2.79 6.01 2.61 2.69 4.94 3.08 2.74 6.05 2.86 2.86 6.12
М 9 – between row intercrop 2.29 2.32 3.22 2.35 2.30 3.25 2.66 2.22 3.43 2.28 2.27 3.07
MH – between row intercrop 2.40 2.14 2.88 2.39 2.15 2.89 2.59 2.11 3.02 2.02 2.04 2.20
Sd 1.0266 1.6722 1.8897 1.2637
Gd 0.5 2.237 3.645 4.119 2.754
48
f 30.8976 24.9451 16.0754 48.4594
ʻFreedomʼ
WA without intercrop (st) 3.02 3.05 7.35 2.90 3.40 8.77 3.02 3.10 7.59 2.62 3.26 7.29
WGP without intercrop 3.28 3.02 7.83 2.75 3.18 7.28 3.35 3.05 8.15 3.12 3.38 9.51
WA with intercrop 3.35 3.19 8.92 3.10 3.28 8,73 3.68 3.04 8.90 3.08 3.49 9,82
WGP with intercrop 3.08 3.12 7.84 2.70 3.18 7.14 3.35 3.06 8.21 3.08 3.40 9.32
ММ 106 – in row intercrop 3.05 2.79 6.21 2.65 2.88 5.75 3.30 2.70 6.29 3.00 3.14 7.74
М 9 – between row intercrop 2.75 2.16 3.36 2.50 2.26 3.34 2.78 1.95 2.77 2.92 2.49 4.74
MH – between row intercrop 2.59 2.27 3.49 2.46 2.20 3.12 2.76 2.10 3.18 2.72 2.46 4.31
Sd 1.0259 1.2636 0.9618 1.2418
Gd 0.5 2.318 2.855 2.173 2.806
f 0.8864 1.0298 0.5139 1.4218
The difference in tree yield at ʻFlorinaʼ cv. The lowest value is the ʻFreedomʼ cv. grafted
under the influence of seed rootstocks is 33.37 on WGP.
kg and ʻFreedomʼ cv. 2.43 kg. Compared to The production costs required to grow one
ʻFlorinaʼ, the ʻFreedomʼ yields are lower with apple plantation from the studied combinations
25 kg/tree on WA rootstocks and 60.97 kg/tree ranged from 1900 euro/ha at ʻFreedomʼ on
on WGP. In the variants with intercrops, better WGP to 2817 euro/ha at ʻFreedomʼ on WGP
results have been obtained with WA rootstock with intercrops MM 106 and M9 (Figure 2).
for ʻFlorinaʼ - 68.73 kg/tree, and for ʻFreedomʼ
with WGP - 60.60 kg/tree.
Using vegetative rootstocks the best yielding is
ʻFreedomʼ grafted on MM 106, followed by
ʻFlorinaʼ on MM 106, with yield difference of
7.6 kg. The lowest average yield per tree was
obtained at ʻFlorinaʼ grafted on MH - 18.4 kg.
In the studied variety-rootstock combinations,
the average fruit weight of both types of seed
rootstocks was not significantly influenced by
the variety and the rootstock and ranged of 91
- 126 g. In trees on vegetative rootstocks, it is
Figure 2. Production costs and net income, euro/ha
larger for ʻFlorinaʼ than for ʻFreedomʼ.
The highest percentage of extra quality of fruit
from ʻFlorinaʼ on WA- 82.97 %, followed by Differences in costs are a result of the number
ʻFreedomʼ on WA - 76.86 %, and with the of trees per hectare, the quantity of the average
lowest ʻFreedomʼ on MH - 21.27 %. On both yield and the associated costs of harvesting
seed rootstocks the quantity of extra-quality additional fruit production.
ʻFlorinaʼ fruit is higher in the variants with Quantity of the average yields and the gross
intercrops. The same trend has not been output obtained have a significant impact on
established for ʻFreedomʼ variety. At all the amount of net income. In the ʻFlorinaʼ cv.,
vegetative rootstocks on ʻFlorinaʼ more than 50 the most effective combination is WA
% of the quality fruits are obtained, while in rootstock with intercrop in rows on MM 106,
ʻFreedomʼ variant only on M9 rootstock. and between rows on MH. For `Freedom`cv.,
The received gross production of variants of better results are obtained using WGP with
ʻFlorinaʼ cv. has the highest value in the intercrops on M9 rootstock, indicating that it is
combination of wild apple with intercrop in the necessary to choose suitable rootstocks for the
row on MM 106 and between rows on MH - individual varieties.
12402 euro/ha. In this indicator ʻFreedomʼ cv.
is grafted on a WGP rootstock with intercrop in CONCLUSIONS
row on MM106 and between row on M9 -
12065 euro/ha (Figure 1). Trees of both varieties on seed rootstock have a
larger stem diameter than those of MM 106,
M9 and MH. Difference was found for both
types of seed rootstocks in variants with and
without intercrops. The thicker tree stem is
explained by the growth force of the seed
rootstocks, which induces a stronger thickening
of the tree stem of the used grafts.
Trees of the ʻFlorinaʼ cv.on different types of
rootstocks have a larger crown volume than
those of ʻFreedomʼ cv.
The production costs required to grow one
hectare of apple plantation from the
Figure 1. Gross production, euro/ha combinations studied range from 1900 euro/ha
49
to 2817 euro/ha. Cost differences are the result REFERENCES
of the number of trees per hectare, the quantity
of the average yield and the associated costs of Dyankov D., 1995. Influence of planting distances on the
harvesting additional fruit production. development of young trees from some summer apple
varieties. Plant Sciences, No 3, 71-74.
In ʻFlorinaʼ cv., the most effective combination Domozetov D., Radomirska I., 2009. Productive
of a rootstock wild apple with in row intercrops behaviour of apple variety-rootstock combinations,
on MM 106, and between rows on MH. cultivated as intercropping plantation. Plant Sciences,
For ʻFreedomʼ cv., better results are obtained 46, 17-20.
using a WGP with intercrop on M9 rootstocks, Fett M., Waquil P., 2001. Analise economica de sistemas
de cultivo de macieiras. Revista Brasileria de
indicating that it is necessary to select suitable Fruticultura, Brazil, 23 (1): 69-73
rootstocks for the individual varieties. Kiprijanovski M., Ristevski B., Arsov T., Gjamovski V.,
2009. Influence of planting distance to the vegetative
ACKNOWLEDGEMENTS growth and bearing of 'Jonagold' apple cultivar on
'MM106' rootstock. Acta Hortic. 825, 453-458.
Mac an Saoir S., Cross G., Mansfield J., Ward F., 2014.
The article is published in collaboration with Millennium planting density trial of 'Bramley's
Project BG05M2OP001-2.009-0034 “Support seedling' apple on M.9 and M.27 rootstocks 2000-
for the development of scientific capacity at the 2012. Acta Hortic. 1058, 613-617.
University of Forestry” financed by the Platon I., Jakab Z.S., Stănică F., 2014. Effect of planting
Operational Program “Science and education system on two apple varieties cultivated in the North-
eastern part of Romania. Acta Hortic. 1058, 181-184.
for intelligent growth”, co-funded by the
European Union through the European
Structural and Investment Funds.
50
STUDIES ON THE BEHAVIOR OF SOME ELITE

AND VARIETIES OF PEAR IN THE POLLINATION PROCESS
Mihaela (ERCULESCU) STOICA, Dorel HOZA, Lenuţa Constantina CHIRA,

Constantin PĂUN, Ligia ION

59 Mărăşti Blvd, District 1, Bucharest, Romania
Abstract
Fungal plant pathogens belonging to the genus Venturia cause damaging scab diseases of members of the Rosaceae. In
terms of economic impact, the most important of these are Venturia inaequalis, which infects apple, and Venturia
pirina, which is a pathogen of European pear. Given that Venturia fungi colonise the sub-cuticular space without
penetrating plant cells, it is assumed that effectors that contribute to virulence and determination of host range will be
secreted into this plant-pathogen interface. The use of resistance varieties in the pollination process is an important
way to obtain varieties with genetic resistance to disease. In this paper were used as mother genitors some selections
from Pyrus serotina (‘9/34-94’, ‘20/1-91’ and ‘5/104-84’) with genetic resistance to diseases and pests and as father
genitors (pollen) two valuable varieties of the European assortment (‘Williams’, ‘Beuré Bosc’) and ‘Cristal’ cv. -
Romanian pear variety registered in 2009 at Research Station for Fruit Growing Voinesti, Dambovita. The selection ‘9
/ 34-94’ performed the best results in the pollination process in crossing process with ‘Williams’ and ‘Cristal’ cvs., with
21.7%, respectively 32.11% fruit set.
Key words: pear, resistance, pathogen, scab.
INTRODUCTION difficult to classify them to particular race.

Understanding the mechanisms of Venturia
The Venturia pathogen might have originated pathogenesis and intricacies of its interaction
in Asia and from there spread to Europe and with apple should provide important insights
recently to other apple growing countries. for developing new strategies to combat the
Hypervariability and evolution of strains that disease (Jha G. et al., 2009). The whole
have overcome host resistance are attributed to genome mutagenesis screen should be initiated
the ability of Venturia to recombine its genetic to identify key virulence factors. The
material every year (Parisi L. et al., 1993).The availability of standardized transformation
conidia and conidiophores, together, give a methodologies in V. inaequalis will facilitate
characteristic velvety appearance to the young such efforts (Tenzer I. et al., 1997).
lesions of scab. The conidia of Venturia are
capable of adhesion and germination on MATERIALS AND METHODS
nonhost plants such as Pyrus communis,
however further development to establish Plant material evaluated included:
infections occurs only on the host plants Pollen from pear cultivated varieties like
(Chevalier M. et al., 2004). The isolates of V. ‘Williams’, ‘Cristal’, ‘Beurré Bosc’, was
inaequalis are hypervariable and exhibit collected in spring 2016.
differential pathogenicity on apple cultivars The resistance selections ‘9/34-94’, ‘20/1-91’,
(known as differential hosts). Based upon such ‘5/104-84’ were used in the controlled crossing
differences, the pathogen has been categorized process as mother genitors.
into eight physiological races (MacHardy W.E., The scab source for inoculum was procured
1996; Bénaouf G. et al., 2000; Bus V.G.M. et from the collection of the Research Station for
al., 2005). However, some of the isolates of Fruit Growing Voinesti, Romania.
Venturia are capable of growing on two These breeding progenies will be characterized
different differential hosts and hence it is after screening infections tests. Infection tests
51
will be performed in greenhouse conditions - the combination C4 of 58 pollinated flowers,
according to Chevalier et al. (1991) will be 4 fruits were bound, with a fruit set percentage
used for selection of resistant plants. Mixtures of 6.89%.
of isolates will be used for plantlet inoculation.
Seedlings will be spraying with a conidial Table 1.The percentage of fruits set
suspension of Venturia inaequalis CKE.
Seedlings will be incubating for 48 hours at
Combinations
nated flowers
No. of fruits
No. of polli
harvested
Genitors
obtained
18°C and 100% relative humidity. Disease
No. crt.
No. of
No. of
♀x♂
seeds
fruits
set
symptoms will be evaluated macroscopically
after 21 days of cultivation in a greenhouse.
Seedlings will be dividing into 5 classes. Plants 9/34-94 x
1. C1 450 98 95 650
in class 0 were without symptoms of infection. ‘Williams’
Plants of class 4 had lesions with full 2. C2
9/34-94 x
273 88 83 535
sporulation. ‘Cristal’
20/1-91 x
3. C3 ‘Untoasa 70 5 5 30
RESULTS AND DISCUSSIONS Bosc’
5/104-84 x
4. C4 ‘Untoasa 58 4 4 32
In order to plan an efficient breeding program Bosc’
to obtain cultivars resistant to pear scab, it is
important to know the genetic control of this In the Figure 2, it is noted that in the hybrid
resistance. Although there is controversy about combination, C1% of the linked fruit did not
the genetic control of the resistance to pear produced hybrid fruit, and in the hybrid
scab, all authors consider that resistance could combination C2 is noted 6% of the linked fruit
be transmitted from resistant progenitors to did not produce hybrid fruit. At C3 and C4
offspring. However, the descendants from combinations, the linked fruits had a 100%
crosses between susceptible and resistant success rate.
cultivars segregated in a complex way.
FRUIT BINDING
SEXUAL HYBRIDATION
No. of binding fruits No.of hybrids fruits
No. of pollinated flowers
No. of hybrids seeds
450
273
650
535
70
58
98
95
88
83
32
30
5
5
4
4
C1 C 2 C3 C4
C 1 C 2 C3 C4
Figure 1. Number of pollinated flowers
In the Table 1 and Figure 1 we observed that Figure 2. The fruits after pollination
the most value combination is C2 with From Table 2 and Figure 3 and 4 we see a
32.23%, percentage of fruit set, followed by the success of 44% for Hybrid Combination C1,
combination C1 with 21.77% percentage of the 31% for Hybrid Combination C2, and for
fruit set: Hybrid C3 we have a success rate of 83% and
- the combination C1 of 450 pollinated flowers, for C4 we have a 100% success rate.
98 fruits were bound, with a fruit set percentage The resistant maternal progenitor (ˋ9/34-94ˋ,
of 21.77% ˋ5/104-84ˋ, ˋ20/1-91ˋ) was able to transmit the
- the combination C2 of 274 pollinated flowers, scab resistance to the descendants, in
88 fruits were bound, with a fruit set percentage agreement with previous results observed by
of 32.23% other authors (Chevalier M., 2004).
- the combination C3 of 70 pollinated flowers,
5 fruits were bound, with a fruit set percentage
of 7.14%
52
along with sub-cuticular pH of such leaves are
speculated to play a role in governing such
resistance. A breakdown of ontogenic
resistance revealed by restored growth of the
pathogen is observed in the old senescing
leaves of apple. Detailed studies are needed to
elucidate the functionality of such resistance
and understand its breakdown mechanism.
Figure 3. Hybrids seeds
Germinated seeds and hybrid

seedlings
100 88
62
50 27 27 20 20
18 15
0
C1 C2 C3 C4
Figure 5. Aspects of pollinations
germinated seeds and hybrid seedlings
No.hybrid seedlings CONCLUSIONS
Figure 4. The percentage of germinated seeds and It might be evident from this review that
number of hybrid seedlings Venturia. Sp. is an important plant pathogen
because it causes huge economic losses and
Table 2.The percentage of germinated seeds and number
also has a very interesting lifestyle. It is an
of hybrid seedlings
appropriate time to sequence whole genome of
the pathogen. The availability of genome
Combinations
No. of seeded
Percentage of
combinations
sequence will not only stimulate research in the

No. hybrid
seedlings
No. crt.
rooting
Hybrid
seeds
field of Venturia pear interactions and

contribute to the basic understanding of this
pathosystem but can also revolutionize the
9/34-94 x understanding of pathogenesis of other obligate
1. C1 62 27 44%
‘Williams’ pathogens. Understanding the mechanisms of
9/34-94 x Venturia pathogenesis and intricacies of its
2. C2 88 27 31%
‘Cristal’
interaction with apple should provide important
20/1-91 x
3. C3
‘Untoasa Bosc’
18 15 83% insights for developing new strategies to
5/104-84 x combat the disease. The whole genome
4. C4 20 20 100%
‘Untoasa Bosc’ mutagenesis screen should be initiated to
identify key virulence factors. The availability
During the course of coevolution, pear has of standardized transformation methodologies
evolved mechanisms to prevent the severity of in V. inaequalis will facilitate such efforts.
scab. The matured leaves of pear demonstrate The resistant elite progenitors (9/34-94, 5/104-
ontogenic resistance because of which the 84, 20/1-91) are an important step on the pear
pathogen growth is suppressed immediately breeding program.The scab conidia could
after cuticle penetration and appearance of germinate on pear leaves undergoing defense
disease symptom gets delayed. The responses, formation of primary hyphae is
strengthened cell wall and cuticular membrane delayed and growth of subcuticular stroma is
53
suppressed resulting in reduced conidiation. Chevalier M., Bernard C., Tellier M. et al., 2004. Host
Phenolic produced in response to V. inaequalis and non-host interaction of Venturia inaequalis and
Venturia pirina on Pyrus communis and Malus x
infections in pear are known to inhibit pathogen domestica. Acta Horticulturae; 663: 205–208.
growth and are ascribed to be associated with Jha G., Thakur K., Thakur P., 2009. The Venturia apple
defense mechanisms of scab resistant cultivars. pathosystem: pathogenicity pathosystem mechanisms
and plant defense responses. J. Biomed. Biotechnol.
REFERENCES 680- 160.
MacHardy W.E., 1996. Apple Scab, Biology,
Bénaouf G., Parisi L., 2000. Genetics of host-pathogen Epidemiology, and Management. St. Paul, Minn,
relationships between Venturia inaequalis races 6 and USA: APS.
7 and Malus species. Phytopathology; 90 (3): 236– Parisi L., Lespinasse Y., Guilaumes J., Kruger J., 1993:
242. A new race of Venturia inaequalis virulent to apples
Bus V.G.M., Laurens F.N.D., Van de Weg W.E. et al., with resistance to the Vf gene. Phytopathology, 83:
2005. The Vh8 locus of a new gene-for-gene 533–537.
interaction between Venturia inaequalis and the wild Tenzer I., Gessler C., 1997. Subdivision and genetic
apple Malus sieversii is closely linked to the Vh2 structure of four populations of Venturia inaequalis
locus in Malus pumila R12740-7A. New Phytologist. in Switzerland. Eur. J. Plant Pathol., 103: 565–571.
166 (3): 1035–1049.
54
RESEARCH ON THE ISOLATION OF GENOMIC DNA

FROM OLD APPLE VARIETIES
Ligia ION

Abstract
The apple scab disease has probably evolved over a long time along with the apples. The disease is caused by the
fungus Venturia inaequalis (Cke.) Wint, anamorph Spilocaea pomi Fr. The aim of this study were the isolation and
quantification the genomic DNA on old Romanian varieties in order to select the most important to them for the Marker
Assisted Selections (MAS) on apple trees. This paper presents the results of DNA isolation by exploring local
populations of apple like ‘Prescurate’, ‘Seghese’, ‘Vieşti’ and ‘Kniş’ apples old varieties, that are well adapted to the
conditions in Romania and that are un interesting genetic potential for resistance to scab. Only the old apple varieties
were used in the study. The implementation of marker selection depends on the quality and quantity of isolated genomic
DNA. The results of the quantification performed following genomic DNA isolation show a large variability in the
amount of DNA in each old apple variety. The DNA concentration in apple varieties both parents and some descendants
have shown that the values are higher in parents than hybrids, with the highest value for ‘Turnu’ variety with 179.1 ng /
μl, followed by ‘Calvil alb’ variety with 161.1 ng / μl, then ‘Varga’ variety with 155.0 ng / μl.
Key words: scab, varieties, apple, resistance.
INTRODUCTION breakdown of this type of resistance has been

found in the literature; thus, ontogenic
Apple (Malus x domestica Borkh.) is one of the resistance is considered durable (MacHardy et
most cultivated fruit crops in temperate al., 2001). Goethe (1887) and Aderhold (1900)
climates. The major constraint of apple are believed to have been the first researchers
cultivation is the apple scab, a fungal disease noticing age-related resistance in the Malus-
caused by Venturia inaequalis, which can lead Venturia pathosystem.
to important crop losses if not properly The authors observed a decrease of leaf
controlled (Holb et al., 2003). susceptibility with increasing tissue age. Nearly
The 17 major resistance genes in apple (Rvi1 to three decades later, Keitt and Jones (1926)
Rvi17) against V. inaequalis have been found showed an increase in incubation period and a
(Bus et al., 2011). decrease of disease severity by increasing leaf
Only Rvi6 (previously Vf from Malus age.
floribunda 821) has been extensively used for Following these observations, many researches
resistance breeding to date (Gessler et al., have been carried out on Malus-Venturia
2012). interaction during leaf infection. Gessler and
Since the breakdown of the Riv6 gene in the Stumm (1984), Li and Xu (2002), and Gusberti
early nineties (Parisi et al., 1993), new breeding et al. (2012) showed that the fungus grew faster
programmes have started to investigate other in young leaves compared to old ones.
resistance genes for future resistance breeding The first unfurled and expanding leaf is
(Gygax et al., 2004; Gaecia-Ruiz and Murphy, considered susceptible to the apple scab
2001). disease, while the fifth leaf (starting from the
An increase in resistance with increasing apple top of the shoot) is considered fully resistant
leaf age (ontogenic resistance) has been (MacHardy, 1996).
observed in all apple genotypes and is known Disease resistance mechanisms during tissue
to act against all known Venturia inaequalis ontogenesis have been studied in different plant
strains. Up to know, no report of the pathogen systems and some factors have been
55
suggested to be correlated to the observed age- with the FP 120 Fast-Prep machine (Bio 101
related resistance. Savant Instruments Inc., Qbiogene, France) for
Among them, the most important appears to be 30 s at a speed of 5.5 m s-1 with an intermediate
chemical compounds such as salicylic acid immersion in liquid nitrogen between the two
(Kus et al., 2002; Hugot et al., 1999) and grinding steps. Total RNA Isolation System
pathogenesis-related proteins (Wyatt et al., (Promega Corporation, USA) and column
1991), physiological barriers like the cuticle purified following the manufacturer’s instruct-
(Peries, 1962), lenticels (Kennelly et al., 2005), tions.
restricted phloem movement (Garcia-Ruiz and After RNA isolation and quality assessment,
Murphy, 2001) or a limiting nutritional samples were stored at -80°C until cDNA
substrate for fungal infection (Juen and library construction and transcriptomic assay
Hwang, 1991). the each population were carried out according
However, since a different mechanism for age- to conditions specified in Zhebentyayeva et al.,
related resistance is described in each crop 2003.
plant, much work remains to unveil the
mechanism underlying this type of resistance in RESULTS AND DISCUSSIONS
other plants. In apple, several aspects have been
investigated in order to unveil the nature of The results on the concentration in genomic
ontogenic resistance. DNA of the studied apple varieties showed that
Physiological barriers like the cuticle and the highest concentration was recorded in
papillae (Stadler, 1988), were not correlated to ‘Turnu’ variety with 179.1 ng / μl, followed by
the age-related resistance. the ‘Calvil alb’ variety with values of 161.1 ng
The objectives of this study were the isolation / μl, then ‘Varga’ variety with 155.0 ng / μl,
and quantification the genomic DNA on old compared to 12.6 ng / μl ‘Rosu Marin’, follo-
Romanian varieties in order to select the most wed by ‘Sălciu’ with 51.2 ng / μl (Table 1).
important to them for the Marker assisted
Selections on apple trees. Table1. DNA concentration in apple varieties
MATERIALS AND METHODS No. Cultivars Conc Ra- Ra- Absor-

DNA pport pport bance
(ng/µl) 260/ 260/ x260
Plant materials 280 230
The old Romanian apples genotypes 1 Prescurate 121.3 2.21 0.23 2.425
(‘Prescurate’, ‘Gurguiate’, ‘Viești’, ‘Kniș’, ‘Turnu’, 2 Turnu 179.1 2.11 0.29 3.582
‘Moharat’) were tested for their ability to 3 Sălciu 51.2 2.50 0.14 1.028
support the artificial infection to scab. All this 4 Venchi 76.9 2.30 0.15 1.537
material was inoculated (by spreeding) with 5 Gurguiate 114.6 1.88 0.60 2.291.
6 Iridium 55.2 2.54 0.12 1.104
fiels races of Venturia inaequalis. 7 Calvilalb 161.1 2.16 0.31 3.222
DNA isolation and SSR analysis 8 Vânători 48 87.2 2.23 0.17 1.744
Genomic DNA was isolated from fresh apple 9 Varga 155.0 2.04 0.29 3.099
leaves using the hexadecyltrimethylammonium 10 Renet 143.7 1.35 0.56 2.874
bromide (CTAB) protocol described by. DNA Portocaliu
concentrations were measured by a NanoDrop. 11 Costat de 46.8 2.00 0.454
Albești
Working of dilutions of genomic DNA at 100 12 Andrifișer 97.8 2.04 5.57 1.957
ng/μl in TE buffer (pH 8.0) were prepared for 13 Roșu Marin 12.6 2.38
analysis. Fresh leaf tissue (<100 mg) was 14 Sângeriu 66.5 1.91 4.55 1.330
sampled from inoculated and uninoculated leaf 15 Roșu de 59.6 2.04 11.83 1.192
samples at 72 and 96 hpi and collected in 2 ml Cluj
Eppendorf tubes (Eppendorf, Germany),
previously prepared with 5 to 10, 2 mm sterile The results of the quantification performed
glass beads, immersed in liquid nitrogen following genomic DNA isolation show a large
immediately after sampling and stored at -80°C variability in the amount of DNA in each old
until processing. Tissues were ground twice apple variety (Figures 1-4).
56
and precipitation of proteins with organic
solvents such as phenol and chloroform;
Following centrifugation, the precipitated
proteins can be separated from the DNA, which
remains in the supernatant.
The implement of resistance older apple
varieties of Romanian origin could be a
promising way for a viable breeding program in
Romania.
First step in this work was the identification
and collection (from different parts of the
country) and evaluation an important number
Figure 1. Apple DNA isolation
of old local varieties (Ion L. et al., 2016)
DNA Concentration Absorbanta 260

200 4
100 2
0 0
Figure 2. DNA concentration on apple cultivars Figure 4. Apple DNA for absorbance 260
DNA purification involves the removing from Using DNA markers of resistance to V.
the lysate all components except DNA, inaequalis will be based on the polymerase
dividing the DNA, that is, separating the chain reaction (PCR).
different DNA species into distinct categories. Recovery in the Romanian breeding program
The two aspects are not necessarily constituted the old local varieties, best suited to the
in separate events. climatic conditions of Romania was used like a
In fact, DNA purification can still begin at the natural source of resistance to pathogen attacks.
stage of cell lysing.
CONCLUSIONS
During three years of field-controlled

pollination for new hybrids, other sources of
resistance such as some old apple varieties,
such as ‘Amelie Wacsman’, ‘Gurguiate’, ‘No
Name’, ‘Mohorât‘, ‘Nobile de Geoagiu‘,
‘Plocsay’s favorite‘ etc.
The final number of signatures was relatively
low.
Regarding the DNA concentration in apple
varieties both parents and some descendants
have shown that the values are higher in
parents than hybrids, with the highest value for
Figure 3. Aspects of DNA isolation
‘Turnu’ variety with 179.1 ng / μl, followed by
Removal (after lysate formation) of all ‘Calvil alb’ variety with 161.1 ng/μl, then
biomolecules and contaminants of DNA is ‘Varga’ variety with 155.0 ng / μl.
accomplished by several methods: denaturation
57
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58
EFFECTS OF GRAFTING COMBINATIONS ON THE FRUIT
QUALITY FOR THE PINOVA APPLE TREE
Radu BĂRĂSCU1,2, Dorel HOZA1, Ligia ION1, Gheorghița HOZA1

1
Faculty of Horticulture, 59 Mărăşti Blvd, 011464, Bucharest, Romania;
2
Vâlcea Plant Nursery, 343 A, Mihăeşti, Vâlcea County, Romania;
[email protected]

Abstract
The quality of apple fruits is influenced by variety and within each variety by the rootstock and by the culture
technology applied. The research presented in this paper highlighted the influence of the rootstock on the fruit quality.
The experiment was conducted during 2016-2017 in the Vâlcea plant nursery, in Romania, as a comparative study for
the ʻPinovaʼ variety with several rootstocks (M9, B9, M20, Pi80, M106), including variants with grafting interstems
(B9/A2, B9/M111). The size of the fruit was larger for the trees grafted on the rootstock B9 with the interstem M111,
while the firmness was positively influenced by the rootstocks M9 and B9/A2. The content of soluble dry substance was
favourably influenced by the rootstocks M20, B9 and Pi 80, while the titratable acidity had higher values for the fruits
produced by the trees grafted on M106 and M9/M111. The total anthocyanins content was higher for the fruits obtained
from the trees grafted on the rootstock B9 with the interstems M111 and A2.
Key words: fruit weight, interstem, rootstock, soil management.
INTRODUCTION standard rootstocks and interstocks are used in

order to decrease the vigour (Webster, 1995).
The apple tree is one of the most important The interstock influences the quality of fruits
fruit-growing specie cultivated in Romania due and the colour grade (Vercammen et al., 2007)
to its economic value and significant produc- and induces precocity (early fruiting) (Webster
tion. Fresh apples are highly appreciated for et al., 1995).
consumption throughout the whole year due to The length of the interstock influences the fruit
their healthy effects on the human body but can production.
also be industrially processed in order to obtain The one with a higher length leads to a higher
various products based on pulp or juice. productivity value and balances growth with
The quality of fruits depends on a number of fructification (Di Vaio, 2009).
factors from which the following can be The research conducted by Samad et al., (1999)
mentioned: the variety, the culture technology, with various dwarf rootstocks used did not
the age of the trees, the pruning used (different highlight any significant differences of fruit
training system) (Lord et al., 1985; Dudu et al., weight in correlation with the rootstocks used.
2015; D’ Abrosca et al., 2017). In order to observe the way in which quality
Apple growers are constantly focused on and quantity of fruits are influenced by the
making the apple production intensive with the rootstocks and interstocks used a study was
aim of reducing the height of trees and the conducted using 13 grafting combinations at
manual labour costs. ‘Pinova’, a scab resistant apple variety, dwarf
Making the production intensive is possible by and semi-dwarf rootstocks and grafting
grafting trees on dwarf rootstocks, particularly interstocks to reduce the vigour.
M9, but those trees need a supporting system.
Also, dwarf rootstocks are more sensitive to
MATERIALS AND METHODS
draught in comparison to standard rootstocks
(Zhou et al., 2016). The research was conducted in the Valcea area
In order to renounce the supporting systems during 2016-2017, in a ‘Pinova’ apple orchard
and to ensure a better soil stability of trees, established in 2015 with the planting distance
59
of 3.6 m x 1.25 m and a number of 2222 used and the results were expressed in kg/cm2
trees/ha. (Saei, 2011).
In order to highlight the influence of the Soluble solids were determined from blueberry
grafting interstock, two lengths were used: 30 juice (Saei, 2011; Oltenacu, 2015), with the
and 40 m respectively. refractive device Kruss DR301-95 (% Brix).
The soil was maintained either worked or The titratable acidity was determined by
grassy. Researchers used 13 grafting combina- titration with 0.1N NaOH to pH 8.1 (Saei,
tions, simple or with interstock, and for each 2011). For titration with 0.1 N NaOH the
combination they chose 9 trees divided in 3 automatic titrator TitroLine easy was used. The
repetitions. results were expressed in g citric acid/100g of
The following experimental variants resulted: fresh weight.
V1 – Pinova/M106 – control; Total anthocyanins content was measured with
V2 – Pinova/M9; spectrophotometrically at wavelength λ = 540
V3 – Pinova/M20; nm (Bărăscu et al., 2016), after an adapted
V4 – Pinova/Pi 80; method. The extracts were filtered under
V5 – Pinova/B9; vacuum and completed up to 50 ml volume.
V6 – Pinova/B9/MM111, interstock 30 cm, The results were calculated using the formula:
worked soil; Total anthocyanins = DO540 x F, where DO540
V7 – Pinova/B9/MM111, interstock 30 cm, is the absorbance and factor F = 11.16. The
grassy soil; total anthocyanins content was expressed in
V8 – Pinova/B9/MM111, interstock 30 cm, mg/100 g of fresh weight.
buried; All determinations described above were
V9 – Pinova/B9/MM111, interstock 40 cm, performed with Specord 210 Plus spectropho-
worked soil; tometer. The preliminary data registered were
V10 – Pinova/B9/A2, interstock 30 cm, grassy statistically interpreted using the method of
soil; variant analysis for probability of 5%, 1% and
V11 – Pinova/B9/A2, interstock 30 cm, worked 0.1 %.
soil;
V12 – Pinova/B9/A2, interstock 40 cm, grassy RESULTS AND DISCUSSIONS
soil;
V13 - Pinova/B9/A2 interstock 40 cm, buried, The average production of fruits per tree for the
grassy soil. first 2 years of fructification (2nd and 3rd year of
For combinations with interstock the influence life) was moderate but different for the experi-
of deep planting was tested, the interstock mental variants (Table 1).
being planted as well. Approximately half of the variants had a
The maintenance technology applied in the relatively equal productivity.
orchard was the standard one used at high- A lower production was registered at the
density apple culture. grafting combinations M20, MM111+B9
At harvest, the data registered regarded the interstock with the length of 30 cm, buried and
production per tree and per hectare and average A2+B9 interstock of 40 cm.
samples of 15 fruits were collected from each The best production for ʻPinovaʼ resulted from
grafting combination on which physical and the combination MM111+B9 interstock of 30
chemical determinations were carried out. cm and worked soil. Statistically, the highest
The dry matter and water content of the production was registered at variants V6 and
samples were determined by oven drying for 24 V7, very significant in comparison to the
hours at 105oC using a UN110 Memmert oven, average one.
method used also by Delian (2011), Corollaro On the other hand, the lowest was at variants
(2014), Muresan (2014). To determine the fruit V3, V10 and V11, significantly negative.
firmness an electronic penetrometer TR was
60
Table 1. Fruit production and average fruit weight at the ʻPinovaʼ apple variety
Variant Production Average fruit weight
kg/tree t/ha g Std
V1 10.0oo 22.22 161.47*** 8.22
V2 11.9 * 26.44 137.41000 7.24
V3 8.5ooo 18.88 156.32*** 6.53
V4 11.3 N 25.11 128.56000 9.20
V5 12.0 ** 26.66 151.62*** 7.13
V6 12.7 *** 28.22 104.41000 8.37
V7 12.3 *** 27.33 161.47*** 6.57
V8 11.9 * 26.44 138.26000 4.36
V9 12.1 ** 26.88 194.52*** 8.62
V10 9.0ooo 19.99 141.91000 5.34
V11 9.5ooo 21.11 114.57000 4.87
V12 11.2 N 24.88 157.70*** 6.58
V13 11.6 N 25.77 169.04*** 4.69
Average- control 11.16 24.61 147.48 4.21
LSD 5% 0.66 0.81
LSD 1% 0.90 1.10
LSD 0.1% 1.21 1.47
* - significant values for the 5% probability; ** - significant values for the 1% probability; *** - significant values for the 0.1% probability.
Average fruit weight was influenced The fruit diameter influenced fruit weight, even
dramatically by the variants used. Thus, the though the variation limit was small (Table 2).
biggest fruits were obtained at V9, over 194 g/ The correlation index between the average
fruit, followed by V13 with 169 g /fruit and weight and fruit diameter was of r = -0.4881
variants V1 and V7 with over 161 g/fruit. The (Figure 1). The grafting combination also
smallest fruits were obtained at V6 with only influenced the fruit firmness. The strongest
104 g/fruit and V11 with roughly 114g/fruit. It fruits were obtained by V2 with 10.24 kg/cm2
is worth mentioning that the first two variants followed by V11 with 9.14 kg/cm2. Lower
with big fruits were the ones with grafting values were noted at V3 and V7 with 7.29
interstock on standard rootstocks. Statistically, kg/cm2 and 7.33 kg/cm2, respectively. All other
the variants were divided in 2 groups: one variants had intermediate values (Table 2).
under average, significantly distinctive and the Statistically, values over 8.60 kg/cm2 were
other above the average, significantly positive. significantly positive and the ones under 7.92
kg/cm2 were significantly negative.
y = 16,068 - 0,1052 x
Correlation: r = -0,4881
10,5
10,0
9,5
Diameter, mm
9,0
8,5
8,0
7,5
7,0
66 68 70 72 74 76 78 80 82
Weight, g 0,95 Conf.Int.
Figure 1. Correlation between fruit weight and fruit diameter
61
Table 2. Diameter and firmness of fruits at the ʻPinovaʼ apple variety grafted on various rootstocks
Variant Fruit diameter Firmness

mm Std kg/cm-2 Std
V1 75.00** 7.07 7.92ooo 0.49
V2 68.33ooo 5.77 10.24*** 1.39
V3 73.33 N 2.89 7.29ooo 0.39
V4 68.00ooo 3.54 8.30 N 0.66
V5 73.33 N 5.77 8.86*** 0.86
V6 75.00** 0.58 8.65*** 0.44
V7 71.67 N 5.77 7.33ooo 0.31
V8 71.67 N 2.89 8.18 o 1.10
V9 80.00*** 0.58 7.69ooo 0.43
V10 75.00** 5.00 8.60** 0.71
V11 68.00ooo 3.54 9.14*** 0.75
V12 75.00** 5.00 8.55** 0.75
V13 78.00 N 3.54 7.91ooo 1.07
Average 73.26 4.00 8.36 0.72
LSD 5% 1.40 - 0.14 -
LSD 1% 1.90 - 0.19 -
LSD 0.1% 2.55 - 0.25 -
* - significant values for the 5% probability; ** - significant values for the 1% probability; *** - significant values for the 0.1% probability.
The water and the total dry substance content and V13 the difference was significantly
were less affected as opposed to the physical negative in contrast to the average one.
parameters of fruits (Table 3). The lowest Glucides formulated in °Brix accumulated
content of water was observed at V11 of more at fruits from V3, reaching the maximum
roughly 78.25%, and the highest at fruits from value (19 °Brix) and it was noted statistically as
V13, of 81.40%. The content of total dry very significant. Lower values were obtained at
substance was complementary to the one of V7 and V8, under 15.5 °Brix. The content of
water. Statistically at V6, V9 and V11 the ash was between 0.21% at V2 and 0.44% at
difference was significantly positive in V11 without a visible correlation with the total
comparison to the average and at V7, V8, V12 dry substance and the content of glucides.
Table 3. Some biochemical parameters of fruits from the ʻPinovaʼ variety grafted on various rootstocks
Variant Water Total dry substance Glucides Ash

% % (°Brix) %
V1 80.69 19.31 N 16.6 N 0.37
V2 79.53 20.47** 17.27 N 0.21
V3 80.08 19.92 N 19*** 0.27
V4 80.14 19.86 N 17.4 N 0.38
V5 80.34 19.66 N 15.8 o 0.27
V6 79.04 20.96*** 16.82 N 0.24
V7 83.48 16.52ooo 14.07 ooo 0.20
V8 81.33 18.67ooo 15.52oo 0.28
V9 78.91 21.09*** 17.46 N 0.31
V10 80.21 19.79 N 16.65 N 0.26
V11 78.25 21.75*** 17.5 N 0.44
V12 81.27 18.73ooo 15.63 o 0.25
V13 81.40 18.60ooo 15.47 N 0.28
Average- Control 80.36 19.64 16.55 0.29
LSD 5% 0.46 0.81
LSD 1% 0.62 1.10
LSD 0.1% 0.84 1.47
* - significant values for the 5% probability; ** - significant values for the 1% probability; ***- significant values for the 0.1% probability.
62
Table 4. Titratable acidity and antocyanin content at fruits from the ʻPinovaʼ variety grafted on various rootstocks
Variant Titratable acidity Anthocyanin
g/100 g f.w. Std mg/100g f.w. Std
V1 0.379 0.001 0.763 0.037
V2 0.330 0.005 0.143 0.063
V3 0.296 0.016 0.706 0.052
V4 0.318 0.004 0.576 0.070
V5 0.348 0.012 0.773 0.060
V6 0.307 0.016 0.537 0.020
V7 0.186 0.001 0.848 0.034
V8 0.298 0.003 1.151 0.033
V9 0.369 0.004 0.603 0.870
V10 0.362 0.006 0.478 0.051
V11 0.342 0.002 0.591 0.057
V12 0.346 0.007 1.064 0.013
V13 0.270 0.005 0.673 0.085
Average - Control 0.319 0.006 0.685 0.111
Fruit acidity oscillated from simple to double, REFERENCES

being weaker at V7, with roughly 0,186 g/ 100
g f.w. and maximum at V1, with 0.379 g/100 g Bărăscu R., Hoza D., Bezdadea-Cătuneanu I., Năftănăilă
f.w. All other variants registered intermediate M., Albulescu A., 2016. Preliminary research
regarding the influence of grafting interstock and soil
values (Table 4). The highest quantity of maintenance on fruit quality for ʻPinovaʼ Variety.
anthocyanins was registered at fruits from V8 Agriculture and Agricultural Science Procedia 10:
and V12 with over 1060 mg/100 g f.w and the 167-171.
smallest values were at fruits from V2 with Corollaro M.L., Aprea E., Endrizzi I., Betta E., Demattè
only 0.143 mg/100 g f.w. M.L., Charles M., Bergamaschi M., Costa F., Biasioli
F., Grappadelli L.C., Gasperi F., 2014. A combined
sensory-instrumental tool for apple quality
CONCLUSIONS evaluation. Postharvest Biology and Technology 96:
135-144.
The present study proved how the grafting D’Abrosca B., Scognamiglio M., Corrada L., Chiocchia
combinations influenced the size and quality of I., Petriccione M., 2017. Evaluation of different
training system on Annurca apple fruits revelated by
fruits of the ʻPinovaʼ variety. agronomical, qualitative and NMR based
Generally, fruits of better quality were obtained metabolomic approaches. Food Chemistry, 222(1):
at combinations were a grafting interstock was 18-27.
used. Interestingly, the most used rootstock, the Delian E., Petre V., Burzo I., Bădulescu L., Hoza D.,
M9, yielded well and produced firm fruits with 2011. Total phenols and nutrients composition
aspects of some apple cultivars and new studied
a high content of dry substance but they were breeding creations lines grown in Voineşti area –
small, weakly coloured and with few minerals. Romania. Romanian Biotechnological Letters vol. 16
Good colours were obtained at V8 and V12 and (6): 165-170.
a high content of dry substance, over 21%, was Di Vaio C., Cirillo C., Buccheri M., Limongelli F.,
registered at V9 and V11. 2009). Effect of interstock (M9 and M27) on
vegetative growth and yield on apple trees (cv.
The worked soil determined a better growth of Anurca). Scientia Horticulturae 119 (3): 270-274.
fruits but it did not register other correlations Dudu G., Cimpeanu M.S., Jerca O.I., 2015. The
for the other indicators observed. The grassy influence of bio-fertilization on production level of
soil assured a better colouration. some harvested apple cultures (Vf) in an intensive
The interstock of 30 cm induced a slight system in the southeast of Romania. Scientific
Papers, Series B, Horticulture, LIX: 43-46.
growth of the fruit size and a satisfactory Lord W.J., Greene D.W., Damon R.A., Baker J.H., 1985.
accumulation of soluble dry substance in Effects on stem piece and rootstock combinations on
comparison to the interstock of 40 cm. growth, leaf mineral concentration, yield and fruit
Deep planting of trees, including of the quality of Empire apples trees. J. Am. Soc. Hortic.
interstock actuated a slight growth of fruit size, Sci. 110 (3): 422-425.
Militaru M., Butac M., Sumedrea D., Chitu E., 2015.
a good firmness and a better colouration. Effect of Metaxenia on the fruit quality of scab
63
resistant apple varieties. Agriculture and Agricultural Samad A, Niel D.N., Khan Z.U., 1999. Effect of
Science Procedia, 6: 151-156. interstock bridge grafting (M9, dwarfing and same
Mureşan E., Muste S., Borşa A., Vlaic R., Mureşan V., cultivars cutting) on vegetative, reproductive growth
2014. Evalution of physical-chemical indexes, sugars, and carbohydrate composition of mature apple trees.
pigments and phenolic compounds of fruits from Scientia Horticulturae 79 (1-2):23-38.
three apple varieties at the end of storage period. Vercammen I., van Daele G., Gomand A., 2007. Can
Bulletin UASVM Food Science and Technology 71 fruit size and colouring of Ionagold be improved by
(1): 45-50. an interstock? Acta Hortic. 832: 165-170.
Oltenacu N., Lascăr E., 2015. Capacity of maintaining Webster A.D., 1995. Rootstock and interstock effect on
the apples quality, in fresh condition-case study. deciduous fruit tree vigour precocity and yield
Scientific Papers, Series Management, Economic productivity. N.Z.J. Crop Hort. Sci., 23: 373-382.
Engineering in Agriculture and Rural Development Zhou B., Sun J., Liu S.Z., Jin W.M., Wei Q., 2016.
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Saei A., Tustin D., Zamani Z., Talaie A., Hall A., 2011. temperatures. A study on physiological features and
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64
PHYSICAL AND CHEMICAL PARAMETERS OF THE FRUIT IN FOUR

PRUNUS DOMESTICA LOCAL POPULATIONS FROM BUZĂU COUNTY
Daniel Constantin POTOR, Aurora DOBRIN, Mihaela Ioana GEORGESCU, Dorel HOZA

Abstract
The four local populations of Prunus domestica, T1, T2, T3, respectively T4, selected for observations and
measurements are distinguished by a series of particular attributes of fruit: one of the four populations matures their
fruits early in August while the remaining populations are ripening during September; the endocarp is not adherent to
the T3 population, whereas the remainder has different degrees of adhesion; the average weight of fruit varies clearly
from species to species, and the ratio between fruit weight and fruit diameter is relatively close for the 3 populations;
the sugar content, with one exception, is close to the average of 15.95, and the dry matter content is also variable
depending on the population.
Key words: Prunus domestica, fruit, physical parameters, chemical parameters.
INTRODUCTION related to local environmental factors, are

valuable information for fruit growers when
Among the fruit species cultivated in our they want to broaden their range of varieties.
country, the plum is on the first place both for By their content in antioxidant substances,
the cultivated area and for the fruit yield along with vitamin C and minerals like
obtained (Coman et al., 2012, calcium, potassium or magnesium
http://www.fao.org/). (https://ndb.nal.usda.gov/), the plum fruits have
The pedo-climatic factors can influence the a significant nutritional role in our diet.
pomological, physical and biochemical Researches on the physical and chemical
proprieties of the local biotypes or genotypes of characteristics of the local varieties of plum
the cultivated fruit tree species (Ionica et al., will contribute to the desired objective, the
2013; Iordanescu and Costea, 2014). improvement of the cultivar assortment (Botu
It is important to have information about these et al., 2012).
particularities, useful to the breeding or Following this direction, our study presents the
growing processes, or to enlarge the assortment morphological and chemical characteristics of
of local fresh, conditioning or preserved fruits four local plum varieties from Buzău County.
(Vitanova et al., 2004; Okatan et al., 2017).
There are different researches on fruits MATERIALS AND METHODS
proprieties from acquainted cultivars or local
varieties, such as the nutritional characteristics The fruits of four local populations in
of local cultivars (Rop et al., 2009), the changes Pătârlagele (Buzău County) (Potor et al., 2017)
that occur in their physical or chemical were collected in full maturity in 2017.
characteristic during the fruits ripening or For their characterization the following
keeping (Usenik et al., 2008; Miletic et al., physical and chemical parameters were used:
2012; Oltenacu and Oltenacu, 2014), the diameter, weight, respiratory intensity, the dry
chemical compounds and their contribution to matter contents, titratable acidity, soluble solid
the fruit aroma, color (Usenik et al., 2009; Pino and total anthocyanins content.
and Quijano, 2012) or to the antioxidant
activity (Donovan et al., 1998); these features,
65
where DO530540 is absorbance at wavelength
λ = 530540 nm and factor F = 11.16 and
expressed in mg/100g plant material (Bezdadea
Cătuneanu et al., 2017).
The titratable acidity was determined according
to AOAC official method 942.15 and Saad et
al., 2014: 10 g of the sample were diluted with
50 ml of water and titrated with 0.1 N sodium
hydroxide to pH 8.1. The formula for
calculation is:
ͳͲͲ
Figure 1. Fruit of T3 population ܶ݅‫ ݕݐ݈ܾ݅݀݅ܿܽ݁ܽݐܽݎݐ‬ൌ

V = volume of titrant; N = normality of titrant;
The four populations are distinct from the C= Citric acid equivalent; m = mass of the
moment of fruit maturation and the degree of sample.
adherence of the mesocarp, as follows:
- the T1 population - the mesocarp partially The analysis was performed with TitroLine
adherent; early ripe period, in the second easy titrator.
decade of August; 0.1 is the normality of NaOH (N), 0.064 is the
- T2 population - adherent mesocarp; the ripe conversion factor for citric acid, V is the
period situated in the first decade of September; volume of NaOH used (mL) and m is the mass
- T3 population - non-adherent mesocarp of plum sample used (g).
(figure 1); the ripe period situated in the first The results were expressed in mg citric
decade of September, too; acid/100 g fresh sample
- T4 population - adherent mesocarp; the latest Total soluble solids (TSS) were determined
ripe period between the 4 populations, at the from plum juice (Bezdadea Cătuneanu et al.,
end of September. 2017), with Kruss DR301-95 Digital Handheld
Fresh fruits were used for physical and Refractometer and the results were expressed in
chemical analysis. % Brix.
The diameter average value was determined by
measuring of 20 fruits / population with the RESULTS AND DISCUSSIONS
fruit caliber; the results were expressed in mm.
The average weight was determined by Diameter, weight and dry matter content
weighing 30 fruits / population with the
Table 1. Fruits diameter, weight and dry matter content
Partner-PS 600 R2 technical balance, the result to the T1-T4 Pătârlagele local populations
being expressed in g.
Dry matter content
Respiratory intensity, based on the CO2 Population Diameter (mm) Weight (g)
(%)
measurement of the plant material and T1 28,82 ± 1.01 15,32 ± 2.05 16,99
expressed in mg CO2 / kg-1h-1, was determined T2
T3
33,41 ± 2.40
44,29 ± 3.50
18,94 ± 3.52
45,50 ± 7.64
27,05
18,24
with the CO2 analyzer. Determination of the T4 33,06 ± 1.48 21,14 ± 1.72 21,71
dry matter content was achieved by weighing
the fresh vegetable material and drying at Data of Table 1 and Figure 1 show that the T3
105°C with the moisture analyzer MAC 50 population form the largest fruits in size and
PARTNER, the result being expressed as a weight, but with a content in dry matter lower,
percentage. 18.24%, relative to the T2 and T4 populations.
For the total anthocyanin content, the adapted The T1 population has small fruits with a low
method was used after Giusti et al., 2001, dry matter content compared to the rest of the
respectively, the extraction in acidified populations.
methanol with 1% hydrochloric acid and The ratio between dry matter content and fruit
spectrophotometric dosing at the wavelength of diameter is obviously higher in T1 and T3
530 nm. Results were calculated based on the populations, while in T2 and T4 populations
formula: Total anthocyanins = DO530540 x F, the differences are smaller (Figure 2).
66
Comparing the data in the table 1 with those while T2 have the highest value of the
from Ionică et al., 2013, it can be noticed that 3 populations studied: 25.30%.
of the 4 populations, namely T1, T2 and T4, T1 and T2 populations have the values of dry
form smaller fruit, the average of the weight matter and total soluble solids content very
between 15.32 g and 21.14 g, are outside of the close, the T3 population is showing a small
average weight range of the fruit from the difference of the two components, while to the
mentioned paper work. T4 population the dry matter content is
obviously higher than that in soluble solids
60 (Figure 2).
50
40
30
20
10
0
weight (g) diameter (mm) dry matter content
T1 T2 T3 T4 (%)
Figure 2. Acid citric, Brix and dry matter content
Figure 1. Physical parameters and dry matter

content of fruits of the T1-T4 populations The higher value of the citric acid content of
the T1 and T3 populations (Table 2)
With an average weight of 45.50 g, the fruits of corresponds to a low content of total soluble
the T3 population account for the highest value solids; in the fruits of the T4 population, the
among the local populations of Pătârlagele; the citric acid content is small, similar to the
dry matter content for T1, T3, T4, ranging from content of total soluble solids, while in the
16.99% to 21.71%, is according with the results fruits of the T2 population the content of total
obtained by Ionică et al.; the fruits of the T2 soluble solids is high and the value of the citric
population, with an average of the dry matter acid is at the medium level comparative to the
content of 27.05%, are above the values recor- rest of populations (Table 2, Figure 2).
ded by the other populations (table 1, figure 1). With the exception of the T4 population where
the difference between the dry matter content
Titratable acidity, soluble solids and dry and the total soluble solids content is
matter content significant, to the rest of the populations the
The citric acid content ranges from 0.11 mg / ratio between citric acid content and dry matter
100 g fresh sample to T4 and 0.81 mg / 100 g content is similar to the total soluble solids
fresh sample at T1 (Table 2). content (Figure 2).
Table 2. Titratable acidity, total soluble solids Respiratory intensity and dry matter
and dry matter content of fruits of the
T1-T4 Pătârlagele local populations
content
Titratable acidity Total soluble Table 3. Respiratory intensity and dry matter content of
Dry matter
Population (mg citric acid solids
/100 g) (% Brix)
content ( %) fruits of the T1-T4 Pătârlagele local populations
T1 0.81 ± 0,089 15.78 ± 1.91 16,99 Respiratory intensity
T2 0,45 ± 0,010 25.30 ± 2.76 27,05 Population Dry matter content (%)
(mg CO2/kg-1h-1)
T3 0,67 ± 0,028 15.87 ± 3.64 18,24 T1 149.60 ± 29.43 16.99
T4 0,11 ± 0,004 15.95 ± 1.28 21,71 T2 64.85 ± 10.93 27.05
T3 104.66 ± 27.66 18.24
The content of total soluble substances is very T4 61.90 ± 7.94 21.71
close to 16% in the T1, T3 and T4 populations,
67
The respiratory intensity (Table 3) is between High respiratory intensity is found in T1 and
61.90 mg CO2 / kg-1h-1 (T4) and 149.60 mg T3 populations.
CO2 / kg-1h-1 (T1). Populations T2 and T3 have In the fruits of the four Pătârlagele populations
an intensity of fruit respiration approx. 2 times the content of anthocyanins is relatively
smaller than T4. reduced.
From Figure 3 it can be seen that the two
populations with less of the respiratory REFERENCES
intensity have the higher dry substance content.
Bezdadea Cătuneanu I., Bădulescu L., Dobrin A., Stan
200 A., Hoza D., 2017. The influence of storage in
150
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70
ʻHARLAYNEʼ Х ʻHARCOTʼ – PERSPECTIVE CROSSBREED

FOR COMBINING GOOD FRUIT QUALITY AND RESISTANCE
TO PLUM POX VIRUS
Marieta NESHEVA, Valentina BOZHKOVA, Snezhana MILUSHEVA
Fruit Growing Institute, 12 Ostromila Street, Plovdiv, Bulgaria
Abstract
Most apricot breeding programs aim to obtain genotypes combining good fruit qualities and resistance to Plum pox
virus (PPV). Using a combination between field observations and Marker Assisted Selection improves the efficiency of
the breeding process. The study was conducted at a breeding orchard in Fruit Growing Institute - Plovdiv, Bulgaria
and describes progenies from the hybrid family ʻHarlayne’ х ‘Harcot’, genotyped for resistance to PPV. Fruit ripening
period, fruit biometry, total soluble solids (TSS), sensory analyses, fruit ground and over color were observed. The fruit
ripening period of the most of the hybrids is after Harcot, the fruits were classified from small to large size and 82% of
them surpass the TSS content of the parental cultivars. According to the sensory evaluation the hybridˈs fruits were
highly scored (5.07-7.67). All of them were with orange ground color of varying intensity. The fruit over color was from
10 to 80 %. Two of the studied hybrids were selected and grafted on rootstock for final evaluation. A lot of the other
hybrids had desirable traits and the breeding process with them will continue in F2 for improving their fruit quality.
Key words: fruit breeding; hybrids; Plum pox virus; pomological traits; Prunus armeniaca L.
INTRODUCTION most devastating disease and a major limiting

factor for the apricot cultivation. The selection
Most apricot breeding programs point at of resistant genotypes requires a lot of time and
genotypes combining good fruit qualities and significantly slows down the whole breeding
resistance to different pests. The commercial process. Therefore, a number of scientists have
value of apricots depends entirely on the fruit focused their efforts on developing molecular
quality. It is determined by the combination of markers associated with genes of PPV
physical and chemical fruit characteristics such resistance (Abernathy et al., 2004). The
as appearance, firmness, taste and aroma development of Marker Assisted Selection
(Velisek and Cejpek, 2007). These (MAS) effectively complements plant breeding
characteristics are genetically highly variable process (Singh B.D., Singh A.K., 2015) and
and their phenotypic expression is influenced gives us an opportunity to make it shorter by
by the environmental conditions in the year of doing a proper selection at the very early stages
cultivation (Dirlewanger et al., 1999). The of it. Our study describes field observations of
knowledge of the ways of inheritance of fruit genotyped for PPV resistance hybrid family
quality traits would result in a higher efficiency ʻHarlayneʼ х ʻHarcotʼ and its aim is to
of the breeding process and would help the investigate the way of inheritance of the fruit
choice of genitors. These traits can be quality traits and evaluate the progeny.
improved by the breeding process and there is a
great diversity of genetic resources for them MATERIALS AND METHODS
(Krška et al., 2006a). Most of the fruit quality
traits are under polygenic control and are This research was conducted at a breeding
quantitatively inherited (Salazar et al., 2013). orchard of Fruit Growing Institute - Plovdiv,
The conventional breeding is successful in Bulgaria. The hybrids were obtained by the
developing cultivars with improved fruit methods of conventional fruit breeding and
quality but the same cannot be said for the planted in 2011. In 2014 the progeny was
disease resistance. Plum pox virus (PPV) is the genotyped for PPV resistance within the work
71
on project MARS (7 FP- Collaborative project RESULTS AND DISCUSSIONS
nr. 613654). This hybrid family consists 153
seedlings. In the present study, the hybrids that Seventeen of the hybrids fruited in the three
fruited consistently over the three years (2015, consistently years. Their ripening period started
2016 and 2017) are described. An average in the second half of June and it’s duration in
sample of fruits was taken and biometric data 2015 was 18, in 2016 - 12 and in 2017 - 20
was measured with Mitutoyo 500-196-30 days. In 2016 the ripening period was shorter
Digimatic Absolute Caliper 150 mm. than in the other two years probably because
Total soluble solid content (Brix0) in juice this trait is strongly influenced by the climatic
using a handheld Sper Scientific 300019 factor (Milošević et al, 2010). During the three
Digital Refractometer was determined. For the years, the same trend was observed - fruit
descriptive characteristics of the fruits, UPOV ripening time is genetically variable trait and in
(2007) and IBPGR (1984) descriptors were the progeny, there are hybrids which fruits
used. Sensory evaluation was done by a group ripen earlier or later than both parental
of trained consumers. cultivars. According to Audergon et al. (2011),
For statistical data processing, Duncan's this is due to the genetic background of the
multiple range test at P≤0.05 through IBM parents which has a strong influence on the
SPSS Statistics 19 was used. inheritance of the fruit ripening time.
June 2015 July 2015 June 2016 July 2016 June 2017 July 2017
15 20 25 30 5 10 15 20 25 30 5 10 15 20 25 30 5 10
Harcot 1 1
2 1 1
1 1 Harcot
4 1 1
3 Harcot 1
Harlayne 2 1
2 Harlayne Harlayne
3 5 2
1 3 1
1 2 3
1 2
3
1
Figure1. Fruit ripening date in 2015, 2016 and 2017

Different digits show the number of hybrids ripened on that date
On figure 1 it is noticeable that most of the and probably for that reason, there is such
hybrids ripened later than ʻHarcotʼ and diversity in the hybrid family.
ʻHarlayneʼ. In our previous study with the
Table 1. Fruit size categories according
crossbreed ʻModestoʼ х ʻHarcotʼ, the biggest to IBPGR descriptor
group of hybrids were with intermediate
Fruit weight Number of
ripening time (Bozhkova and Nesheva, 2016). (g)
Fruit size (IBPGR)
hybrids
In both cases, most of the hybrid fruits ripened < 20 Extremely small 0
later than ʻHarcotʼ cv. 20 - 30 Very small 0
This might means that ʻModestoʼ and 31 - 40 Small 8
ʻHarlayne’ later ripening time is the dominant 41 - 45 Small/medium Harlayne + 3
trait. 46 - 55 Medium 3
These results are in accordance with Nyujtó 56 - 60 Medium/large Harcot + 2
and Banai (1986) proposition for who the late 61 - 70 Large 1
ripening period is dominant. Bassi and Negri 71 - 85 Very large 0
(1991) assume it is under oligogenic control >85 Extremely large 0
72
Fruit weight is a trait of which depend the fruit sorting (Mohsenin, 1980). All three fruit
quality and often the yield. According to the dimensions depend on the cultivar.
IBPGR descriptor, 47% of the fruits of the After Duncan's multiple range test at P≤0.05, it
studied progeny were classified as small size can be said that: according to the data for all
(table 1). Attractive and medium-sized fruits fruit dimensions - length (FL), width (FW) and
are preferred by the producers and consumers thickness (FT) the groups are overlapping and
and also are desired trait in apricot breeding there are hybrids with intermediate phenotype
(Guerriero et al.; 2005). Almost half of the and hybrids that resemble the parental cultivars
studied hybrids (53 %) had small/medium to (table 2).
large sized fruits. Many well-known cultivars Although the clear trend that the fruits in the
are classified as small to medium-sized fruits progeny are getting smaller in size a few
with an average weight up to 50-55 grams hybrids were found to surpass both parental
(Bozhkova and Todorova, 2012). Two of the cultivars: by FL – HH 12-42 and HH 12-26, by
hybrids had medium/large fruits as Harcot and FW – HH 12-19 and HH 12-42, by FT – HH
one surpassed them with its large sized fruits. 12-26, HH 12-42 and HH 12-19 but the
There is a high correlation between the main statistical difference with ʻHarcotʼ is non-
fruit physical characteristics - fruit height, fruit significant. The biggest fruit weight was
width, and thickness and all three of them are recorded for HH 12-42 which is close to the
highly correlated with the fruit weight one measured for ʻHarcotʼ. The larger fruits
(Mratinic et al., 2011). Fruit biometry is had stones with higher weight. However, HH
important when the fruits are intended for 12-42, HH 12-26, and ʻHarcotʼ had very good
processing, especially for their mechanical stone relative share- less than 6.
Table 2. Fruit biometric analysis
Avarage Fruit
Genotype Fruit Length Fruit Width Fruit Thickness Stone weight Relative share %
Weight
HH 13-3 45.03 bcdef 41.39 abcdef 45.36 abcde 48.43 abc 3.06 abcd 6.32 abcd
HH 13-54 43.26 cdef 37.93 cdef 43.64 abcdef 41.89 bcd 3.27 abc 8.10 abc
HH 13-43 41.13 def 37.98 cdef 39.51 ef 35.53 cd 2.27 de 6.36 abcd
HH 12-63 45.74 bcde 40.97 abcdef 45.34 abcde 47.27 abcd 3.07 abcd 6.51 abcd
HH 13-14 38.58 f 37.39 def 39.72 ef 34.60 cd 2.07 e 6.04 bcd
HH 12-26 52.80 a 41.63 abcd 47.72 abc 55.87 ab 2.90 bcde 5.23 d
HH 12-67 47.49 abcd 39.60 abcdef 45.36 abcde 46.49 abcd 2.80 bcde 6.04 bcd
HH 12-42 50.65 ab 45.74 a 49.08 ab 63.09 a 3.57 ab 5.59 cd
HH 12-19 48.08 abc 44.46 ab 49.83 a 59.02 ab 3.87 a 6.56 abcd
HH 13-4 41.06 def 34.54 f 38.58 ef 32.64 cd 2.77 bcde 8.58 ab
HH 13-15 41.24 def 38.44 bcdef 40.88 def 36.45 cd 2.28 de 6.42 abcd
HH 12-62 44.56 bcdef 38.77 bcdef 42.50 bcdef 42.12 bcd 2.77 bcde 6.85 abcd
HH 12-41 43.45 cdef 36.55 def 40.31 def 36.14 cd 2.46 cde 7.30 abcd
HH 12-9 49.63 abc 38.84 bcdef 42.02 cdef 44.40 bcd 2.69 cde 6.08 bcd
HH 12-59 38.46 f 38.40 bcdef 39.60 ef 37.29 cd 2.49 cde 7.14 abcd
HH 12-60 38.80 f 34.86 f 39.49 ef 30.22 d 2.63 cde 8.82 a
HH 12-22 38.99 ef 34.96 ef 38.34 f 32.73 cd 2.53 cde 8.56 ab
Harlayne 43.57 cdef 39.72 abcdef 43.30 abcdef 43.29 bcd 2.67 cde 6.23 abcd
Harcot 49.83 abc 44.12 abc 46.58 abcd 58.44 ab 3.04 abcd 5.19 d
* Mean values followed by different letters within a column are significantly different by Duncan’s multiple range test at P≤0.05.
Measuring the TSS content is a method that can cultivars ʻBerge cotʼ, ʻFlavor cotʼ, ʻLady cotʼ,
quickly give us an idea of the fruit biological ʻTom cotʼ, ʻPerle cotʼ, ʻJenny cotʼ and ʻSweet
value. For 128 apricot cultivars cultivated in cotʼ TSS content ranges from 13.40 to 23.30
Malatya, Turkey TSS range is from 11-26.50 °Brix (Bozhkova and Nesheva, 2016). Total
°Brix (Asma and Ozturk, 2005). For the Soluble Solids (TSS) content is important
73
especially for the dried apricot production. In the present study the lowest TSS content was
High TSS content is associated with high recorded for HH 12-42 (15.5 °Brix) and the
amount of sugars in the apricot fruits and highest - HH 12-22 (21.9 °Brix). Here the trend
increases their quality and the yield of dried is reversed - TSS content increases in the
product (Akin et al., 2008). TSS content grows progeny and 82% of the hybrids outperform
with the fruit ripening, i.e. the highest value both the parental cultivars (table 3).
(Brix0) is found in fully ripened fruits (Xi et al., The differences between ʻHarcotʼ and all
2016). This chemical characteristic is strongly hybrids are non-significant. There is a statis-
influenced by the environmental factors tically significant difference between the other
(Bartolini et al.; 2015). parental cultivar ʻHarlayneʼ and HH 12-22.
Table 3. Fruit appearance and taste qualities
Total Soluble
Relative area of fruit Sensory
Genotype Solids (°Brix) Fruit ground color Sensory score
over color (%) evaluation
(TSS)
HH 13-3 20.07 abcd med. orange 10-30 6.22 Good

HH 13-54 21.47 ab med. orange 80 5.71 Good
HH 12-63 18.8 abcd light to med. orange 10 5.91 Good
HH 13-14 19.97 abcd med. to dark orange 30-40 5.26 Good
HH 12-26 18.2 abcd light to med. orange 50-60 7.67 Very Good
HH 12-67 19.83 abcd light to med. orange 20-50 5.91 Good
HH 12-42 15.5 d med. orange 40-60 6.66 Good
HH 12-19 20 abcd med. orange 60-70 6.33 Good
HH 13-15 21.00 abc med. to dark orange 50 5.71 Good
HH 12-62 15.8 cd med. to dark orange 30 5.53 Good
HH 12-41 16.77 abcd med. orange almost missing 5.11 Good
HH 12-9 18.33 abcd light to med. orange 40-50 5.31 Good
HH 12-59 18.67 abcd med. to dark orange 40-60 5.07 Good
HH 12-60 20.97 abc light to med. orange 10-30 5.74 Good
HH 12-22 21.9 а med. orange 40 5.93 Good
Harlayne 16.47 bcd med. to dark orange 60 6.62 Good
Harcot 17.37 abcd med. to dark orange 10 6.03 Good
Like most of the fruit characteristics, the (Mazza and Miniati, 1993). The intensity and
ground color is genetically determined trait relative area of fruit over color strongly depend
which expression is highly influenced by the on the light, radiation, irrigation and nutrition
environment. Consumers in our country prefer of the trees. The relative area of fruit over color
large fruits, with dark orange ground color and for the studied hybrids ranged from 10 to 80 %.
bright red over color (Bozhkova and Nesheva, More than half of them (65%) have an over
2016). All of the fruits of the studied hybrids color above 40% which gives them very
were with orange ground color which shade attractive appearance. After sensory evaluation,
varies from light to medium and from medium all the 17 hybrids were highly scored (5.07-
to dark orange. The red blush is the most 7.67).
attractive feature of apricots and the larger the Their taste was evaluated as good as both
area it occupies is the more seductive the fruits parental cultivars with already proven qualities.
are. It has a great commercial impact and it is One of the hybrids HH 12-26 is evaluated as
much-desired trait in the breeding programs better than the others with very good taste and
74
score 7.67. Usually, well-informed consumers the stones and fruits of three of the hybrids.
prefer fruits with good taste and when they are Two of them were genotyped as sensitive and
valued by sensory analyzes, taste and aroma are for one of them the data is missing.
of greater importance (Bozhkova and Nesheva,
2016). CONCLUSIONS
Krška et al., (2006b) prove that Harleyne's
resistance is controlled by three independent As a result of the hybrid analyses, for their
complementary dominant genes, and after 10 good fruit qualities and resistance to PPV, HH
years of research, Polak and Kominek (2012) 12-42 and HH 12-19 were grafted on
report it as immune to 6 strains of Plum pox P.cerasifera rootstock and continue to the next
virus. In this breeding program, this cultivar stage of the breeding process. These two
was used as a donor of resistance. Harcot is hybrids were the best ones in the studied
partially resistant to PPV - resistant to strain progeny. The others also have good qualities
PPV - D but it is susceptible to PPV - M and the work with them will continue to F2 for
(Rankovic et al., 1997). Its fruits are large with improving their disadvantages. HH 12-26 also
excellent taste and in this breeding program and has good fruit qualities but on the field, it
much more is used as a donor of good fruit showed symptoms of PPV. The fruit weight of
quality (Karayiannis, 2005). After genotyping the resistant genotypes - HH 13-3 and HH12-22
within the work on project MARS (7 FP- should be improved. Because of the great
Collaborative project nr. 613654) ʻHarlayneʼ heterozygosity and the big number of traits
was found to be resistant. Partial resistance was under polygenic control, picking up two
detected in ʻHarcotʼ. In the progeny in 47% of hybrids out of only 17 is a considerable success
the hybrids were genotyped as resistant, 33% as for the breeding program. Usually, such a result
partially resistant and 20% - sensitive (table 4). is obtained by observing hundreds of hybrids.
Very high percent of the progeny is resistant or
Table 4. Resistance to PPV virus partially resistant to the PPV. That gives us a
reason to believe that the crossbreed ʻHarlayneʼ
Phenotype/PPV х ʻHarcotʼ is very perspective for the apricot
Genotype PPV resistance MAS
symptoms 2017
breeding programs.
HH 13-3 resistant -
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76
EVALUATION OF INTRODUCED PLUM CULTIVARS UNDER

AGROCLIMATIC CONDITIONS OF PLOVDIV REGION, BULGARIA
Valentina BOZHKOVA, Petar SAVOV
Fruit Growing Institute, 4004 Plovdiv, Bulgaria
Corresponding author e-mail: [email protected]
Abstract
The study was carried out in the experimental plantation of the Fruit-Growing Institute - Plovdiv in the period 2014 -
2016 with six German plum cultivars: ‚'Jojo', 'Topstar plus', 'Topgigant plus', 'Toptaste' and 'Tophit plus' compared to
'Stanley' as a standard. The tree volume, trunk diameter, angle of the skeletal branches, annual shoot length growth,
average number of flower buds on spurs and shoots, some phenological characteristics, fruit biometrical analyses,
chemical composition and the sensory characteristics were studied. The biggest tree volume and trunk diameter were
calculated for the cultivars 'Toptaste' and 'Stanley'. The largest angle of skeletal branches was measured on 'Jojo'. The
biggest shoot length was recorded in 'Tophit plus'. The earliest flowering time was observed on 'Jojo' and 'Toptaste', the
latest one on the 'Topstar plus' and 'Tophit plus'. The ripening period of the investigated cultivars was in August and
only for 'Stanley' and 'Tophit plus' in September. According to the biometric data 'Topgigant plus' and 'Tophit plus'
were with the largest fruits in size. The highest sugar content and total soluble solid were found of 'Toptaste's fruits.
The investigated cultivars are suitable to grow under agroclimatic conditions of Plovdiv region.
Key words: plum cultivars, vegetative growth, phenological characteristics, fruit characteristics.
INTRODUCTION processes within the tree canopy, i.e., tree

architecture, is required to optimize the
Plum is a traditional fruit crop in Bulgaria. The growing technologies and especially for the
South Central Region represents 25.6 % of the right choice of training and pruning methods.
total area occupied with plum trees, which is As an answer to this need, some plum cultivars
the first place among the six regions of the were introduced from Germany in the Fruit
country (Agrostatistics, 2016). Growing Institute - Plovdiv.
The main grown cultivar is still 'Stanley', In this study are presented the results of an
because of its good adaptability (Djuvinov V. investigation on some of those cultivars. The
and Vitanova I., 2002). A lot of new cultivars aim is to recommend the best one to the plum
were registered in Europe as tolerant to Plum producers.
pox virus (Sharka disease), but few of them
were accepted by the producers and spread in MATERIAL AND METHODS
the orchards (Jacob, 2002; Blažek and
Pištěková, 2009). The study was carried out in the period 2014-
Till now only the plum cultivar 'Jojo' is known 2016 at the Fruit Growing Institute, Plovdiv.
as resistant to Plum pox virus and in the past The trees of the studied cultivars 'Jojo', 'Topstar
ten years was widespread in the orchards plus', 'Topgigant plus', 'Toptaste' and 'Tophit
(Neumüller et al., 2010). plus' compared to 'Stanley' as a standard were
Unfortunately, it turned out that this cultivar is planted in a collection plantation in 2011 on
susceptible to late spring frosts, which force alluvial-meadow soil at a distance of 4×4 m
Bulgarian producers to look for other cultivars. and grown under non-irrigated conditions and
At the same time, the manipulation of tree without pruning.
architecture is the cornerstone of horticultural Dimensions of minimum five trees were
management. determined to be calculated the tree volume and
According to Costes et al. (2004) an accurate trunk diameter. The angle between the skeletal
knowledge of growth, branching and flowering branches and the central leader was measured
77
in four trees of each cultivar in three conse- growth dynamic is the faster. The other
cutive years, to trace if the angle will be cultivars are in the second group. Among them,
changed during the period of tree growth. the smallest tree volume was registered on
One skeletal branch from four trees per cultivar 'Jojo' and 'Topgigant plus' but statistically it is
was selected for measuring the average annual not proven.
shoot length growth and the average number of Data concerning average trunk diameter shows
flower buds in a spur and a shoot. there are no statistically proven differences. It
The observed phenological characteristics is known that on fruit species the larger angle
included flowering and fruit ripening. Biome- of the skeletal branches is preferred (Vitanov,
trical, chemical and sensory analyses of the 1977). Out of the studied cultivars, the largest
fruits were performed. Data were statistically angle of the skeletal branches was recorded for
processed by Duncan's test (Steele and Torrie, the cultivars 'Jojo' and 'Topgigant plus'.
1980). Nevertheless, it was found that the wood for the
'Topgigant plus' is fragile and very often under
RESULTS AND DISCUSSIONS the weight of the fruits the skeleton branches
are broken.
Data concerning the tree volume and trunk In 'Jojo' and 'Toptaste' the larges angle was
diameter are presented in Table 1. The biggest recorded for the secondary branches. The
tree volume and trunk diameter were calculated dimensions for 'Jojo' are favorable and
for the cultivars 'Toptaste' and 'Stanley'. Their statistically proven.
Table 1. Tree volume and trunk diameter during the period 2014-2016
Tree volume, m3 Trunk diameter, cm
Cultivar 2014 2015 2016 Average 2014 2015 2016 Average

'Jojo' 1.15 1.50 1.91 1.52 b1 14.38 23.00 25.13 20.84 a
'Tophit plus' 1.12 2.09 2.53 1.91 b 15.33 23.33 25.33 21.33 a
'Topgigant plus' 1.20 1.61 2.10 1.64 b 17.25 25.13 27,38 23.25 a
'Topstar plus' 1.25 1.93 2.55 1.91 b 17.00 26.38 28.00 23.79 a
'Toptaste' 2.46 3.05 3.89 3.13 a 21.25 28.75 30.75 26.92 a
'Stanley' 2.77 3.02 3.82 3.20 a 22.50 28.13 30.00 26.88 a
1
Mean values followed by different letters within a column are significantly different by Duncan’s multiple range test at
P≤0.05.
Table 2. Angle of the main and the secondary skeletal branches (°)
Branch angle º (2014 -2016)

2014 2015 2016 Average
Cultivar
Main Secondary Main Secondary Main Secondary Main Secondary
branch branch branch branch branch branch branch branch
angle angle angle angle angle angle angle angle
'Jojo' 66.25 77.50 70.00 76.25 78.00 75.25 71.42 a 76.33 a
'Tophit plus' 40.00 46.25 38.75 38.75 40.25 39.50 39.67 c 41.50 cd
'Topgigant plus' 47.50 44.29 51.25 45.00 55.75 47.00 51.50 b 45.43 c
'Topstar plus' 37.50 37.14 35.00 37.50 38.00 42.00 36.83 c 38.88 d
'Toptaste' 52.50 57.50 43.75 53.75 47.25 55.50 47.83 b 55.58 b
'Stanley' 35.00 42.50 37.50 36.25 40.50 38.25 37.67 c 39.00 d
1
Mean values followed by different letters within a column are significantly different by Duncan’s multiple range test at
P≤0.05.
78
According to Costes et al. (2004) the final size
of trees is a cumulative variable resulting from
the annual shoot length developed each year.
In the studied cultivars, the largest annual
shoot length in 2014 was established in
'Topstar plus' and 'Topgigant plus' (Figure 1).
In 2015 the largest annual shoot length was
reported for 'Tophit plus' and 'Toptaste' and in
the third vegetation for 'Toptaste' and
'Topgigant plus'. In the same year for all
investigated cultivars were obtained the
highest dimensions.
That confirms the research by some authors Figure 1. Annual shoot length per skeletal branch
that shoot growth is strongly influenced by
climatic factors and, above all, rainfall of the studied cultivars varied from 3.26 to
(Seleznyova et al., 2003). The average values 3.96, but the differences between the cultivars
of the studied trait showed that 'Stanley' and are statistically insignificant (Table 3).
'Jojo' had the smallest total annual shoot length The largest number of flower buds on shoots
and 'Tophit plus' had the largest one. The mean was reported in the cultivars 'Jojo' and
number of flower buds in the spurs 'Toptaste' and it is statistically proven.
Table. 3. Number of flower buds on spurs and annual shoots in plum cultivars
On one spur On 1 m of annual shoot

Cultivar
2014 2015 2016 Average 2014 2015 2016 Average
'Jojo' 3.44 3.26 3.74 3.48 a 10.35 28.13 32.45 23.64 a
'Tophit plus' 3.54 3.32 3.56 3.47 a 0.92 12.66 11.38 8.32 b
'Topgigant plus' 3.40 3.28 3.28 3.32 a 9.93 12.13 13.01 11.69 b
'Topstar plus' 3.30 3.30 3.96 3.52 a 19.88 17.98 16.88 18.25 ab
'Toptaste' 3.34 3.38 3.70 3.47 a 20.58 25.18 23.11 22.96 a
'Stanley' 3.48 3.54 3.90 3.64 a 3.86 10.45 9.40 7.90 b
Among the investigated cultivars earlier The stones of the cultivars 'Topgigant plus' and
flowering time was recorded for 'Jojo', and 'Stanley' were above 2 g in weight and in the
'Toptaste'. Two to three days later started to rest varied from 1.52 to 1.87 g, but the diffe-
flower the cultivars 'Stanley', 'Topgigant plus', rences between the cultivars are statistically
'Tophit plus' and 'Topstar plus'. insignificant.
Fruits of the studied cultivars ripen within a The fruits of 'Topgigant plus' were large but
period of one month - from the beginning stones were small and the relative stone to fruit
of August to the beginning of September ratio is low (3.17).
(Figure 2). This data for 'Stanley' is the most unfavorable.
Similar results about the period of ripening of
those cultivars were also obtained by other August September
authors in the Czech Republic and Poland Cultivar 1-10 10-20 20-30 1-10 10-20
(Blazek and Pistekova, 2009). Jojo
According to the data of the biometric analysis Tophit plus
the fruits of 'Topgigant plus' and 'Tophit plus' Topgigant plus
were very large (Figure 3). Topstar plus
The fruits of the other four cultivars were Toptaste
Stanley
medium in size and it was statistically proven
(Table 4). Figure 2. Fruit ripening time
79
Table 4. Fruit biometry ( 2014-2016)
Fruit
Stone Relative share
Cultivar Thickness Fruit weight
Lenght (mm) Width (mm) weight (g) (%)
(mm) (g)
'Stanley' 50.06 ab 35.31 b 37.32 b 38.10 b 2.10 a 5.51 a
'Jojo' 47.55 ab 36.20 b 35.62 b 37.29 b 1.87 a 5.05 ab
'Toptaste' 42.67 c 36.51 b 37.61 b 35.43 b 1.66 a 4.66 abc
'Topgigant Plus' 51.11 a 42.27 a 42.78 a 53.06 a 2.10 a 3.97 bcd
'Topstar Plus' 46.15 bc 38.34 b 37.64 b 38.91 b 1.52 a 3.90 cd
'Tophit plus' 51.44 a 42.20 a 44.36 a 55.20 a 1.72 a 3.17 d
Determining the total soluble solids of the proven. The acid content is low. Only in
fruits is the quickest way to get information 'Topgigant plus' it was 1.75%. For all of the
about the content of the major chemical studed cultivars pH varied within a small
components. The total soluble solids varied range - from 2.99 in 'Topgigant plus' to 3.61 in
from 14.77% in 'Topgigant plus' to 25.90% in 'Stanley'. According to the obtained data, the
'Toptaste' and the differences between them is cultivar 'Toptaste' has the best chemical
significant (Table 5). composition. The fruit quality is a complex of
The data showed that the highest sugar content many different characteristics describing both
was established in the cultivar 'Toptaste'. The external appearance and taste qualities. That is
values for the other cultivars varied from why the sensory evaluation is as important as
8.91% to 10.86% but statistically it was not the chemical analysis.
Table 5. Chemical composition of plum fruits (2014 - 2016)
Total soluble Sugar, % Titratable

Cultivar solids (°Brix) Total Invert Sucrose acidity, % рН
'Stanley' 18.15 b 10.77 b 6.69 bc 4.10 ab 0.82 b 3.61 a
'Jojo' 18.33 b 10.86 b 8.37 a 2.37 b 1.08 b 3.44 ab
'Toptaste' 25.90 а 13.18 a 7.79 ab 4.57 a 1.06 b 3.64 a
'Topgigant Plus' 14.77 c 8.91 b 5.70 c 3.05 ab 1.75 a 2.99 c
'Topstar Plus' 15.43 bc 9.56 b 5.37 c 4.05 ab 1.29 ab 3.07 bc
'Tophit plus' 18.13 b 9.00 b 6.43 c 2.42 b 1.03 b 3.40 ab
The results of the sensory analysis are taste is of the greatest importance for grading
presented in Table 6. The larger fruit size is the cultivars in sensory evaluation.
always preferred not only for plum, but The best taste had 'Toptaste' followed by
generally for all fruits. The difference in the 'Tophit plus' (Figure 5). For the other cultivars
score of this property between the studied this score is not so impressive. The studied
cultivars was just 1.5 points and does not cultivars showed big variations in aroma and
correspond exactly to the established fruit less in sweetness.
weight of the cultivars. In this case, the According to the final evaluation only for
assessment is subjective and depends on the cultivars 'Toptaste' and 'Tophit plus' the
participants of the testing panel. sensory characteristic was excellent. Looking
Compared to the fruit size, the score for the at the values of the chemical analysis we could
fruit shape varied less. Fruit colour together not find a correlation between the chemical
with fruit size contributes to fruit properties and the results of the sensory
attractiveness. Judging by the scores given for evaluation. Similar conclusions have been
fruit coloration, it is obvious that the dark made in our previous studies of plum and
coloured fruits are preferred (Figure 4). apricot (Bozhkova, 2014; Bozhkova and
Taste qualities combine the scores given for Nesheva, 2016).
the texture, taste, aroma and sweetness The
80
Table 6. Sensory evaluation of the investigated plum cultivars fruits
Appearance Taste quality

Fruit Fruit Fruit Total Final
Cultivar size shape colour Texture Taste Aroma Sweetness score evaluation
'Stanley' 7.3 7.3 7.4 6.6 6.5 5.9 6.45 47.4 first class
'Jojo' 6.9 7.3 7.3 7.0 6.6 6.1 6.3 47.5 first class
'Toptaste' 6.4 7.0 6.7 7.5 8.4 7.2 7.8 51.0 excellent
'Topgigant
7.7 7.6 6.7 6.5 5.1 4.9 5.4 44.1 first class
Plus'
'Topstar Plus' 7.8 7.8 7.6 6.8 5.6 5.4 6.0 47.0 first class
'Tophit plus' 8.4 8.1 7.2 7.3 7.1 6.7 6.9 51.8 excellent
Figure 3. Cultivar 'Topgigant plus' Figure 4. Cultivar 'Jojo' Figure 5. Cultivar 'Toptaste'
CONCLUSIONS keep in mind that 'Jojo' is sensitive to late

spring frost.
The biggest tree volume and trunk diameter
were calculated for the cultivars 'Toptaste' and REFERENCES
'Stanley'. The largest angle of skeletal
branches was measured on 'Jojo'. Agrostatistics, Production of fruits - harvest 2016,
The biggest shoot length was recorded in Ministry of Agriculture, Food and Forestry Republic
of Bulgaria.
'Tophit plus', but a more uniform growth rate Blažek J., Pištěková I., 2009. Preliminary evaluation
over the years was established in 'Topgigant results of Top. new plum cultivars in a dense
plus'. It was found that the wood of 'Topgigant planting. Horticultural Science, Prague, 36, 45-54.
plus' is quite fragile. Bozhkova V., 2014. Chemical composition and sensory
The earliest flowering time was observed on evaluation of plum fruits. Trakya University Journal
'Jojo' and 'Toptaste', the latest one on the of Natural Sciences, 15 (1): 31-35.
'Topstar plus' and 'Tophit plus'. Bozhkova V., Nesheva M., 2016. Some results of
evaluation of new-introduced apricot cultivars under
The ripening period of the investigated conditions of Plovdiv region. Agricultural science
cultivars was in August and only for 'Stanley' and technology, v. 8,3: 262-265.
and 'Tophit plus' in the beginning of Costes E., Lauri P.E., Laurens F., Moutier N., Belouin
September. A., Delort F., Legave J.M., Regnard J.L., 2004.
According to the biometric data 'Topgigant Morphological and architectural traits on fruit trees
which could be relevant for genetic studies: a
plus' and 'Tophit plus' had the largest fruits in review. Acta Hortic. (ISHS) 663, 349–356.
size. Djuvinov V., Vitanova I., 2002. Plum production in
The highest sugar content and total soluble Bulgaria. Acta Horticulturae (ISHS) 577, 25-31.
solids were found in the 'Toptaste' fruits. Jacob H.B., 2002. New plum and mirabelles varieties
The investigated cultivars are suitable for out of the breeding work and development in
Geisenheim. Acta Horticulturae (ISHS) 577, 173-
growing under agroclimatic conditions of 176.
Plovdiv region. Neumüller M., Treutter D., Hartman W., 2010.
We do not recommend to producers the Breeding for sharka resistance and high fruit quality
cultivar 'Topgigant plus' and remind them to in European plum (Prunus domestica L.) at
81
Weihenstephan: breeding strategy and selection dwarfing rootstock/interstock combinations. Ann.
tools. Acta Hortic. (ISHS) 874: 221-228. Bot. 91 (6), 665-672.
Seleznyova A.N., Thorp T.G., White M., Tustin S., Steele R., Torrie J., 1980. Principles and procedures of
Costes E., 2003. Application of architectural statistics, McGraw - Hill, New Jork, USA.
analysis and AMAP mod methodology to study Vitanov M., 1977. Genetic Investigations Concerning
dwarfing phenomenon: the branch structure of Prunus domestica L. Cultivar Hybridization,
‘Royal Gala’ apple grafted on dwarfing and non- Publishing House of the Bulgarian Academy of
Science, Chap. VI, 107 - 112.
82
ADVENTITIOUS SHOOT REGENERATION FROM PETIOLE EXPLANTS

IN BLACK CHOKEBERRY (ARONIA MELANOCARPA)
Ionela RUSEA1, Aurel POPESCU2, Valentina ISAC3, Anca Nicoleta ȘUȚAN2, Dorel HOZA1
1
59 Mărăști Blvd, Bucharest, Romania
2
University of Pitești, 1 Târgul din Vale Street, Pitești, Romania
3
Research Institute for Fruit Growing, 402 Mărului Street, Mărăcineni, Romania
Abstract
Due to outstanding nutritional and health benefits, and also to its ornamental value, black chokeberry was gaining
recently high interest from the small fruit growers in Europe. Together with vegetative propagation, in vitro
micropropagation from meristems and adventitious shoots offers suitable methods for the rapid clonal propagation of
new or improved cultivars, to provide sufficient quantities of planting material to the growers and to accelerate the
establishment of large black chokeberry plantings. In this respect, different concentrations of N6-benzylaminopurine
(BAP), dichlorophenoxyacetic acid (2,4-D) and indole butyric acid (IBA) in Murashige and Skoog (MS) and Lee-
Fossard (LF) basic culture media, respectively, were assessed for their effects on adventitious shoot regeneration of the
black chokeberry cultivar 'Nero'. The ability of callus formation and shoot regeneration from petiole segments was
assessed using various combinations of BAP (2.5; 5.0; 10 mg L –1), 2,4-D (0.25; 0.5; 1.0 mg L–1), and IBA (0.25; 0.5;
1.0 mg L–1). Data on callus formation and shoot regeneration were recorded after 60 days of culture. The highest
percentage of black chokeberry petiole explants forming callus (100%) was found in treatments containing a
combination of 2.5 mg L–1 of BAP, 0.25 mg L–1 of 2,4-D, and 0.25 mg L–1 of IBA in MS medium. The only growth
regulators combination which resulted in 100% petiole explants forming callus on both MS and LF media was
5 mg L–1 of BAP, 0.5 mg L–1 of 2,4-D, and 0.5 mg L–1 of IBA. Adventitious shoot regeneration from petiole-derived
callus was high in treatments with 10 mg L–1 and 1.0 mg L–1 IBA, on both MS and LF basic media. Excepting the
cytokinin-auxin combination of 2.5 mg L–1 of BAP, 0.25 mg L–1 of 2,4-D and 0.25 mg L–1 of IBA, shoot regeneration
from petioles of 'Nero' cv. was better on MS medium. However, the best adventitious regeneration and the highest
number of shoots formed per explant ocurred by direct organogenesis. Thus, an average number of 4.3 shoots per
petiole explant was achieved through direct organogenesis on MS medium supplemented with BAP at 5 mg L –1, 0.5 mg
L–1 of 2,4-D, and 0.5 mg L–1 of IBA.
Key words: Aronia, in vitro culture, growth regulators, callus, organogenesis.
INTRODUCTION al., 2010; Ochmian et al., 2012; Rugină et al.,

2012).
Aronia melanocarpa (Michx.) Elliot (black Aronia melanocarpa berries constitute a very
chokeberry), a native North American shrub rich source of numerous substances exerting a
which was naturalized and is well adapted in beneficial impact on health, including mainly
Europe, is an extraordinary medicine plant polyphenols (proanthocyanidins, anthocyanins,
(McKay, 2001). ‘Nero’, ‘Rubina’, ‘Viking’, flavonoids, and phenolic acids), possessing
‘Galicjancka’, ‘Fertödi’, ‘Hugin’, ‘Aron’ and antioxidative, anti-inflammatory, antiviral,
‘Melrom’ are among the most popular varieties anticancer, antiatherosclerotic, hypotensive,
of A. melanocarpa in Europe (Strigl et al., antiplatelet, and antidiabetic properties
1995; Kulling and Rawel, 2008; Walther and (Benvenuti et al., 2004; Slimestad et al., 2005;
Schnell, 2009; Borowska and Brzoska, 2016; Naruszewicz et al., 2007; Jakobek et al., 2012;
Șuțan et al., 2017). They differ from each other Bădescu et al., 2015; Borowska and Brzóska,
in the efficiency of juice extraction, content of 2016; Park et al., 2017). Now it is well known
total polyphenols, anthocyanins and proantho- that black chokeberry possesses one of the
cyanidins, total antioxidative capacity, as well highest antioxidant activities among fruits
as the weight and diameter of the fruit (Rop et (Denev et al., 2012).
83
A. melanocarpa berries are also known to be Recently, Park et al. (2017) reported that A.
rich of cyanidin glycosides (Wiczkowski et al., melanocarpa show beneficial effects against
2010). Therefore, black chokeberries has been hepatic lipid accumulation along with
extensively investigated for their phenolic improvements in body weight, liver functions,
compound content, antioxidant properties and lipid profiles and antioxidant capacity
potential positive influence on the health suggesting the potential therapeutic efficacy of
(Tanaka and Tanaka, 2001; Jakobek et al., its juice on nonalcoholic fatty liver disease (a
2007; Rop et al., 2010; Jakobek et al., 2012; hepatic manifestation of metabolic syndrome).
Litwinczuk, 2013). Aronia phenolics are considered to be also
From the researches on the antioxidant beneficial for cardiovascular health (Wu et al.,
(Kahkonen et al., 2001; Wu et al., 2004; 2017).
Oszmianski and Wojdylo, 2005; Olas et al., Successful plant micropropagation from in
2008; Denev et al., 2012), anti-inflammatory vitro cultured meristems has been reported for
(Zapolska-Downar et al., 2012), hepato- Aronia species, including A. melanocarpa
protective (Kowalczyk et al., 2003; Valcheva- (Brand and Cullina, 1990; Brand and Cullina,
Kuzmanova et al., 2004), cardioprotective (Bell 1992; Petrovic and Jacimovic-Plavsic, 1992;
and Burt, 2007; Naruszewicz et al., 2007), Velchev and Mladenova, 1992; Staniene et al.,
hypotensive and lipid lowering (Hellstrom et 1999; Litwińczuk, 2002; Mahečić, 2009;
al., 2010; Park and Park, 2011), hypoglycaemic Litwinczuk, 2013; Kwak et al., 2015; Șuțan et
and antidiabetic effects (Simeonov et al., 2002; al., 2017) and A. arbutifolia (Kane et al., 1991).
Rugină et al., 2011; Bădescu et al., 2015; However, there is no relevant information on
Banjari et al., 2017), to those on antimutagenic adventitious shoot regeneration using somatic
(Gasiorowski et al., 1997; Duthie et al., 2007) tissue explants in Aronia melanocarpa.
and antitumoral effects (Malik et al., 2003; Adventitious shoot organogenesis and somatic
Bermudez-Soto et al., 2007; Olas et al., 2010; embryogenesis are the basis for implementing
Sharif et al., 2013), or those on protective new genetic variability and biotechnological
action against degenerative diseases, the approaches in woody species, particularly if
scientific literature is rich in information somatic tissues from valuable cultivars are used
highlighting their prophylactic and therapeutic (Silvestri et al., 2016). Regardless of the type
properties, without suggests on any unwanted (direct organogenesis, indirect organogenesis
or side effect of their use (Kokotkiewicz et al., or somatic embryogenesis), regeneration
2010). process is usually influenced by biotic factors
Zielińska-Przyjemska et al. (2007) studied the including genotype, explant type, and abiotic
in vitro effects of Aronia melanocarpa juice on factors such as culture media and
oxidative metabolism and apoptosis of environmental conditions. Although plant cell
neutrophils from obese and non-obese totipotency theoretically enables any of the
individuals, and reported that Aronia juice cells to retain the ability to regenerate whole
exert beneficial effects in cells and may, new plants through organogenesis or somatic
therefore, be useful in the treatment of obesity embryogenesis, the regeneration capacity of
disorders. plant cells usually varies between species,
Bijak et al. (2011) and Sikora et al. (2012) cultivars, and explant types (Ganeshan et al.,
reported the anticoagulant properties of Aronia 2002). Proper regeneration of adventitious
extract, based on the results of their studies, shoots rely on composition of nutrient medium,
which showed significant inhibition of platelet plant grow regulators and types of explant
aggregation after black chokeberry extract (Popescu and Isac, 2000; Isac and Popescu,
administration. 2009).
Experimental data indicate that not only the In the present study, we investigated the ability
fruit but also the leaves of A. melanocarpa and of adventitios shoot regeneration of ‘Nero’, one
their products may be effective means for of the most valuable black chokeberry cultivars
prevention and treatment of the effects of toxic in both central and south-eastern Europe, using
action of some xenobiotics in humans petiole explants, and found that both basic
(Borowska and Brzoska, 2016). culture medium and growth regulators
84
combination and concentration affected The freshly prepared explants were placed
adventitious shoot regeneration. aseptically onto regeneration medium in 100 ml
glass jars (four petiole segments per jar), each
MATERIALS AND METHODS containing 40 ml of sterile medium solidified
with 0.9% plant agar (Duchefa Biochemie).
Petiole segments of about 10 mm in length, Based on our previous experience with
excised from in vitro micropropagated shoots raspberry petiole segments cultured in vitro
of chokeberry cultivar ‘Nero’, were cultured on (Popescu and Isac, 2000), the cut ends of
Murashige and Skoog (MS) (1962) and Lee chokeberry petiole segments were slightly
and Fossard (LF) (1977) basic culture media, deeped into the culture medium in order to
respectively, supplemented with N6- prevent dehydration and promote the nutrients
benzylaminopurine (BAP) in concentration of and growth regulators uptake. The explants
either 2.5, 5.0 or 10 mg L–1, dichloro- from the same donor plantlet were randomly
phenoxyacetic acid (2,4-D) in concentration of distributed in different jars in order to avoid the
either 0.25, 0.5 or 1.0 mg L–1, and indole possible errors in interpretation of results due
butyric acid (IBA) in concentration of either to the differences in physiological state of the
0.25, 0.5 or 1.0 mg L–1 (Table 1). source of explants. Each treatment consisted of
For medium preparation, separate stock 20 petiole segments, in five replications.
solutions of macronutrients and micro-nutrients For all treatments, the petiole segments
were used. Iron was added to the medium as cultured in vitro were subjected to an initial
separate stock solution of ferric sodium salt dark treatment for one week, at 20-22°C.
EDTA (32 mg L–1). BAP and IBA were Subsequently, the cultures were maintained in
disolved in 1N HCl and 1N NaOH, the growth chamber at 23±1°C, under a 16
respectively. Dextrose was used as carbon hours photoperiod of 30 lE m-2 s-1 from cool
source in the culture media (40 g L–1). The pH white fluorescent tubes. Transfer of the
of the culture medium was adjusted to 5.7 with explants to fresh medium was performed every
0.1N KOH before autoclaving for 20 minutes at 4 weeks.
121ιC.
Table 1. Composition of the culture medium used for in vitro adventitious regeneration of shoots
in ‘Nero’ cultivar of A. melanocarpa (Michx.) Elliot
Growth regulators
Experiment Basal medium (mg L–1)
2,4-D IBA BAP
E1 MS 0.25 0.25 2.5
E2 MS 0.5 0.5 5.0
E3 MS 1.0 1.0 10.0
E1 LF 0.25 0.25 2.5
E2 LF 0.5 0.5 5.0
E3 LF 1.0 1.0 10.0
The observations on adventitious shoot the standard analysis of variance (ANOVA)

formation by direct organogenesis or via callus with mean separation by Duncan’s test (p >
were made weekly, and after two months of 0.05).
culture, when regeneration frequency RESULTS AND DISCUSSIONS
(percentage of the petiole explants with at least
one shoot) and number of shoots per explant The in vitro response of black chokeberry
were recorded. For all treatments were petiole explants from cultivar ‘Nero’ was
calculated both the percentage of petiole significantly influenced by both the culture
explants forming shoots and the average medium and concentration of growth regulators
number of shoots per explant. Data for shoot tested. The highest percentage of petiole
regeneration were analyzed for significance by explants forming callus (100%) was found in
85
treatments containing a combination of 2.5 mg al., 2013; Kwiecien et al., 2013; Szopa and
L–1 of BAP, 0.25 mg L–1 of 2,4-D, and 0.25 mg Ekiert, 2014).
L–1 of IBA in Murashige-Skoog medium. We did not find a strong correlation between
The only growth regulators combination which the concentrations of the auxins in the culture
resulted in 100% petiole explants forming calli medium, and type and amount of callus formed
on both Murashige-Skoog and Lee-Fosard by the petiole explants. However, the green
media was 5 mg L–1 of BAP, 0.5 mg L–1 of 2,4- calli derived from petiole explants cultured on
D, and 0.5 mg L–1 of IBA. MS medium containing auxins in smaller
While petiole explants cultured on MS medium concentrations showed a higher ability to form
formed small amounts of callus in all adventitios buds and regenerate shoots by
treatments, only a few on those cultured on LF indirect organogenesis (Figure 1C). Highly
medium were induced to form callus in proliferative petiole-derived calli (Figure 1B)
treatments with IBA and 2,4-D in did not show ability to form adventitious buds.
concentrations of 0.25 mg L–1 and 0.5 mg L–1, Depending on the treatment, shoot regeneration
respectively. Although the presence of two was observed either through direct
different auxins in the culture medium, which organogenesis where the regenerants emerged
is not usual, would be expected to induce a mostly at the cut edge of explants or wound
higher potency of callus formation, a sites (Figure 1 and Figure 2), or indirect
synergistic effect of IBA and 2,4-D was not organogenesis (via callus), where regenerants
obvious. This observation is important in the appeared on the entire surface of the petiole-
context of recent reports showing the derived calli (Figure 1).
production and accumulation of hydroxy- Optimum combination of plant growth
benzoic acids and other biologically active regulators for shoot regeneration was medium-
phenolic acids in shoot and callus cultures of dependent (Table 2).
Aronia melanocarpa (Michx.) Elliott (Szopa et
Table 2. Frequency of adventitious shoot regeneration in vitro by either direct organogenesis
or indirect organogenesis (via callus) from petiole explants of black chokeberry cultivar ‘Nero’
Average number of Average number
Average length Average number
Experimental adventitious shoots of calli formed
of adventitious of shoots regenerated per
variants regenerated per petiole per petiole
shoots (cm) petiole-derived callus
explant explant
E1-MS 0.9±0.41 c 2.23±0.15 ab 1.1±0.20 a 0.6±0.15 b
E2-MS 4.3±0.62 a 2.80±0.48 a 0.9±0.06 ab 1.0±0.00 a
E3-MS 3.4±0.42 ab 2.03±0.60 abc 1.0±0.00 ab 0.9±0.10 a
E1-LF 1.4±0.36 c 0.97±0.19 cd 0.1±0.06 d 0.9±0.06 a
E2-LF 2.1±0.80 bc 0.92±0.12 d 0.4±0.20 cd 1.0±0.05 a
E3-LF 2.6±0.48 bc 1.56±0.21 bcd 0.6±0.17 bc 0.9±0.06 a
*Values presented are mean ± SE. Means followed by the same letter are not significantly different (Duncan test, p>0.05)
The best development of adventitious buds and explants of black chokeberry cultivar ‘Nero’
formation of shoots was achieved on MS cultured onto LF medium, and those of
medium supplemented with 0.1 mg L–1 IBA, adventitious shoots regenerated on MS medium
0.1 mg L–1 2,4-D, and 0.5 mg L–1 BAP. (Table 2). The best adventitious regeneration
Excepting the cytokinin-auxin combination of and the highest number of shoots formed per
2.5 mg L–1 of BAP, 0.25 mg L–1 of 2,4-D and explant ocurred by direct organogenesis
0.25 mg L–1 of IBA, shoot regeneration from (Figure 1 and Figure 2). Thus, an average
petioles of ʻNeroʼ cv. was better on MS number of 4.3 shoots per petiole explant was
medium. However, the statistical analysis achieved through direct organogenesis on MS
showed that there is no difference between the medium supplemented with BAP at 5 mgL–1,
percentages of adventitious shoots regenerated 0.5 mgL–1 of 2,4-D, and 0.5 mg L–1 of IBA
in vitro by direct organogenesis from petiole (Table 2).
86
A B
C D
Figure 1. A-D: Adventitious shoot regeneration of black chokeberry cultivar ‘Nero’

through direct organogenesis from petiole segments
The petiole-derived shoots formed from only a few shoots grew and suppressed the
adventitious buds through direct organogenesis development of the rest. Although a high
were not uniform in appearance (Figure 2), frequency of shoot formation is most often
probably due to interaction between the desired even from the initial tissue explants,
endogenous hormones in the plantlets used as individual shoots rather than clusters are
source of explants and plant growth regulators generally advantageous, because they have a
added in the shoot induction (regeneration) higher vigor (and consequently a better ability
medium. to multiplicate), and also because the thin and
Regardless of the culture medium, when petiole crowded clusters of shoots could not be
explants from chokeberry cultivar ‘Nero’ separated easily for the stage of multiplication.
formed multiple adventitious buds (Figure 1D),
87
A B
C D
Figure 2. A-D: Length and vigour of the adventitious shoots regenerated in vitro by direct organogenesis
from petiole explants of black chokeberry cultivar ‘Nero’
As with micropropagation of Aronia, which is Jacimovic-Plavsic, 1992; Velchev and

considered far more efficient than other Mladenova, 1992; Staniene et al., 1999;
conventional cloning methods like layering or Litwińczuk, 2002; Mahečić, 2009; Litwinczuk,
softwood cuttings (Litwinczuk, 2013), 2013; Șuțan et al., 2017). Even the use of
adventitious shoot regeneration from somatic Woody Plant Medium (WPM) (Lloyd and
tissue explants (e.g. petiole segments) proved McCown, 1980) became frequent with woody
to be effective by direct organogenesis and plants, including Aronia (Kwak et al., 2015;
even from tissue-derived calli (at a lower Chen, 2017), MS medium is a choice with
frequency) on Murashige and Skoog medium, reliable results, supported by many authors,
and much more less effectively on Lee and such as Brand and Cullina (1990; 1992), who
Fossard medium. The effectiveness of MS reported that both MS medium and WPM
medium for the in vitro culture of chokeberry medium supported vigorous shoot proliferation
was emphasized in many published reports in Aronia arbutifolia and A. melanocarpa.
(Brand and Cullina, 1990; Kane et al., 1991; Currently, experiments are underway in our
Brand and Cullina, 1992; Petrovic and laboratory to improve the conditions of in vitro
88
culture of A. melanocarpa, in order to achieve Benvenuti S., Pellati F., Melegari M., Bertelli D., 2004.
higher regeneration percentages per petiole Polyphenols, anthocyanins, ascorbic acid, and radical
scavenging activity of Rubus, Ribes and Aronia.
explant. Journal of Food Science, 69: FCT 164.
Bermúdez-Soto M.J., Tomás-Barberán F.-A., García-
CONCLUSIONS Conesa M.-T., 2007. Stability of polyphenols in
chokeberry (Aronia melanocarpa) subjected to in
We have undertaken the first studies to vitro gastric and pancreatic digestion. Food
Chemistry, 102 (3): 865-874.
investigate the ability of adventitious shoot Bijak M., Bobrowski M., Borowiecka M., Podsedek A.,
regeneration by either direct or indirect Golanski J., Nowak P., 2011. Anticoagulant effect of
organogenesis from somatic tissues of Aronia polyphenols-rich extracts from black chokeberry and
melanocarpa. grape seeds. Fitoterapia, 82: 811-817.
In the current study, shoots were induced by Borowska S., Brzóska M.M., 2016. Chokeberries
(Aronia melanocarpa) and their products as a
both direct and indirect organogenesis from possible means for the prevention and treatment of
petiole segments of in vitro cultured plants of noncommunicable diseases and unfavorable health
A. melanocarpa, cultivar ‘Nero’. effects due to exposure to xenobiotics.
Our results showed the possibility of Comprehensive Reviews in Food Science and food
establishing an effective in vitro adventitious safety, 15 (6): 982-1017.
Brand M.H., Cullina W.G., 1990. Micropropagation of
shoot regeneration system for Aronia
Aronia arbutifolia and A. melanocarpa. Hort
melanocarpa, which holds great promise for Science, 25 (9):1096.
either micropropagation and/or genetic Brand M.H., Cullina W.G., 1992. Micropropagation of
transformation studies in black chokeberry. red and black chokeberry (Aronia spp.). Hort
Also, the results of this investigation may be Science, 27 (1): 81.
Cheng Y., 2017. Tissue culture of Aronia melanocarpa.
useful in optimizing shoot regeneration systems
Protection Forest Science and Technology.
for other cultivars of Aronia melanocarpa, and Denev P.N., Kratchanov C.G., Ciz M., Lojek A.,
also as an alternative allowing rapid mass Kratchanova M.G., 2012. Bioavailability and
propagation of elite genotypes independent antioxidant activity of black chokeberry (Aronia
from seasonal influences. melanocarpa) polyphenols: in vitro and in vivo
evidences and possible mechanisms of action.
Comprehensive Reviews in Food Science and Food
ACKNOWLEDGEMENTS Safety, 11: 471-489.
Duthie S.J., 2007. Berry phytochemicals, genomic
This research was supported by a Doctoral stability and cancer: evidence for chemoprotection at
Research Support grant from the Ministry of several stages in the carcinogenic process. Molecular
National Education. We greatly appreciate the Nutrition & Food Research, 51:665-674.
Ganeshan S., Caswell K.L., Kartha K.K., Chibbar R.N.,
support received through the collaborative 2002. Shoot regeneration and proliferation. In:
work undertaken with the Laboratory of Tissue Khachatourians G.G., McHughen A., Scorza R., Nip
Culture at the Research Institute for Fruit W.K., Hui Y.H. (Eds), Transgenic Plants and Crops.
Growing, and University of Pitești. Marcel Dekker, Inc., New York, 69-84.
Gasiorowski K., Szyba K., Brokos B., Kołaczynska B.,
Jankowiak-Włodarczyk M., Oszmianski J., 1997.
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92
PHYSIOLOGICAL STUDY OF VARIOUS CULTIVARS

OF PUNICA GRANATUM (L.) WITH AN EYE TO ACCLIMATIZATION
TO ROMANIA’S ENVIRONMENTAL CONDITIONS
Ana-Maria PETRE, Aurelia DOBRESCU, Adrian PETICILĂ, Monica Luminița BADEA

59 Mărăști Blvd., District 1, Bucharest, Romania
Abstract
The phytotherapeutic and economic importance given to Punica granatum (L.) in the last decades indicates the rebirth
of this species. In Romania, the species is barely known by farmers and hobby growers, while the pomegranate products
are widely consumed. This research is supposed to ensure the physiological data that will be the basis for the
development of pomegranate crop technology for eight P. granatum cultivars that were introduced for acclimatization
in the experimental fields of University of Agronomic Sciences and Veterinary Medicine of Bucharest. In this study,
physiological processes are described, insisting on photosynthesis, transpiration and respiration in interrelation with
the chlorophyll content, water and dry matter content existing in leaves. The measurements were performed during the
main growth and flowering phenophases, starting late March 2016. The results showed a connection between the
photosynthesis process, leaf dry matter and leaf chlorophyll content, transpiration process and leaf water content,
respiration process and the flowering phenophase (in some cultivars, not in all).
Key words: phenophase, photosynthesis, respiration, transpiration, chlorophyll.
INTRODUCTION have been published, focusing on treatment and

prevention of cancer, cardiovascular disease,
Punica granatum (L) is a fruit-bearing shrub, diabetes, bacterial infections or antibiotic
belonging to the Punicaceae family, having its resistance (Jurenka, 2008).
origins in Middle East. From its origin, Considering lack of water resources and
considered now Iran and Afghanistan, the intensification of abiotic stresses such as
pomegranate spread east to India and China and drought and salinity, importance of pome-
west to Mediterranean countries such as granate has increased in recent years, since this
Turkey, Egypt, Tunisia, Morocco, Greece, Italy species is a tolerant fruit crop and thrives well
and Spain. It is assumed that Spanish under arid and semi-arid climatic conditions
missionaries brought the pomegranate to the (LaRue, 1980; Jamali and Bonyanpour, 2017).
American continent in the 1500’s (Hodgson, The P. granatum shrubs have an effective
1917; LaRue, 1980). control of the plant water status by reducing
In Romania, the shrub can be cropped transpiration due to stomatal closure and
especially in the areas with a warm climate maintaining a consistent maximum stem with
such as Dobrogea and Banat. changes in evaporative demand (Intrigliolo et
Pomegranate is the symbol and heraldic device al., 2011; Parvizi et al., 2016).
of the ancient city of Granada in Spain, from This species can not be cropped only in regions
which the city gets its name. The genus name, with water deficit, but in seasonally wetlands as
Punica, was the Roman name for Carthage, well.
where the best pomegranates were known to The photosynthetic rate, as well as transpiration
grow. Pomegranate is known by the French as and respiration processes depend on various
grenade, the Spanish as granada, and literally parameters like chlorophyll content, leaf water
translates to seeded (“granatus”) apple and dry matter content, abiotic factors
(“pomum”) (Jurenka, 2008). (temperature, light intensity, humidity), plant
In the past decade, numerous studies on the age or plant phenophases (Burzo et al., 2005).
antioxidant, anticarcinogenic, and anti-inflam- The present research, whose results are
matory properties of pomegranate constituents recorded in the writing, aims to thoroughly
93
fathom the physiological and biochemical Photosynthesis as well as transpiration and
particularities of certain P. Granatum cultivars respiration process intensities were determined
of different origins, more precisely to compare with the LCA-4 electronic analyser and
certain physiological parmeteres during the expressed in µmol/CO2/m2/s (photosynthesis
species’ vegetation period, informations that and respiration) and mmol/H2O/m2/s (transpi-
will allow the crop technology acquaintance for ration).
this species’ acclimatization to Romania’s The photosynthetic pigments content was
pedo-climatic conditions. determined using 80% acetone extraction and
by colorimetry at wavelengths of 470 nm, 663
MATERIALS AND METHODS nm and 646 nm. The results were calculated
using the Lichtenthaler & Wellburn’s formula
Plant material was represented by eight P. (1983) and the results obtained expressed in
granatum cultivars: Hicaz, Kandahar, Nikitski mg/100g fresh weight (FW).
ranni, Echen, Mollar, Shahvar, Dolce and The total water and dry matter content were
Local selection cultivar. determined through gravimetric analysis. And
The botanical characterization of the selected expressed in percentages.
P. granatum cultivars was based on
macroscopic (visual) observations made RESULTS AND DISCUSSIONS
directly in the study field.
Thus, the studied plants are characterized as The intensity of the Photosynthesis process
deciduous shrubs, with a gray-brown bark, that in P. granatum leaves during the Growth
grow between 0.7 and 1 meter tall and have 3 Phenophase
to 5 branches with twigs of a length between 3 The maximum photosynthetic rate is recorded,
and 60 cm. for most species, before growth cessation of the
The leaves are simple, petiolated, entire, leaves, when they reached 37 - 90% of their
lanceolate, oposite, coriaceous and have a foliar surface (Burzo et al., 2004).
length of 3 to 7 cm and a width of 1 to 2 cm. From Table 1 data analysis, it is observed that
The plant assortment lies on a 60 m2 surface the photosynthesis process recorded similar
and comprise a total of 13 plants of kindred age values at Echen and Mollar cultivars (8.26
(aprox. 6 years old), originated in Europe and µmol/CO2/m2/s). They carried out the photo-
Middle East, planted at a 2.5 m distance synthesis process with the highest intensities,
between plants and 2.7 m distance between the 1.30 times higher than Hicaz and Dolce, 1.23
rows (also, there is no plant support system) in times higher than Kandahar, 1.94 times higher
the experimental field belonging to Fruit than Shahvar and 1.51 times more intense than
Growing Department of the Faculty of Local selection cultivar.
Horticulture. Table 1. The intensity of the Photosynthesis process in P.
The measurements were performed during the granatum leaves during the Growth Phenophase
main growth and flowering phenophases, on
Light Temperature Photosynthesis
similar leaves belonging to the middle level of Cultivar intensity (°C) intensity
the analyzed plants, starting late March 2016 (µmol/m2/s) (µmol/CO2/m2/s)
(up until late June 2016), 3 times a week - for Hicaz 1245 18.0 6.32
1189 18.1 6.68
the phisyological processes - between 9:00 am Kandahar
Nnikitski 1204 17.7 8.01
and 10:00 a.m, at a light intensity with values ranni
between 1083 and 1586 µmol/CO2/m2/s and an Echen 1153 17.5 8.26
air temperature of 17.5 up to 19.7°C. Mollar 1083 18.6 8.26
Also, the analysis of photosynthetic pigments Shahvar 1073 19.0 4.25
and total water and dry matter content were Dolce 1126 17.7 6.32
completed twice a month, the determination of Local 1119 17.8 5.45
photosynthetic pigments was made using the selection
leaves belonging to the upper level (young

leaves) of the P. granatum plants taken in Nikitski ranni also recorded a high photo-
study. synthetic rate of 8.01 µmol/CO2/m2/s.
94
The Shahvar cultivar, during the growth period, 1.83 times, Kandahar 1.25 times, Nikitski ranni
carried out the photosynthesis process with the 1.21 times, Echen 1.25 times, Mollar 1.09
lowest intensity, compared to the other studied times, Shahvar 1.22 times, Dolce 1.24 times
cultivars, namely 4.25 µmol/CO2/m2/s. and Local selection cultivar 1.44 times.
Of note were Hicaz, Dolce and Kandahar The Hicaz cultivar (with the highest photo-
cultivars, whose photosynthesis processes were synthetic rate) carried out the photosynthesis
carried out with related intensities, the values process with an intensity of 1.38 times higher
being between 6.32 and 6.68 µmol/CO2/m2/s. than Kandahar, 1.19 times higher than Nikitski
Of the obtained data, it was found that the ranni, 1.12 times higher than Echen, 1.28 times
photosynthesis process was influenced by the higher than Mollar, 2.23 times higher than
type of cultivar taken in study, representing an Shahvar, 1.46 times higher than Dolce and 1.29
internal factor that the specialty literature times higher than Local Selection.
signals in modifying dynamics of the
photosynthesis process (Burzo et al. 2005; Total leaf Chlorophyll content of P.
Dobrescu, 2007; Bădulescu, 2016). granatum during the Growth Phenophase
Following the chlorophyll pigment analysis, the
The intensity of the Photosynthesis process highest content in total leaf chlorophyll was
in P. granatum leaves during the Flowering found at Nikitski ranni cultivar, 38.78 mg/100g
Phenophase FW, followed by Echen cultivar, 28.14
During the appearance of the first flowers at mg/100g FW (Table 3).
Hicaz, Kandahar, Nikitski ranni and Local
Selection cultivar, the photosynthesis process Table 3. Total leaf Chlorophyll content of
was carried out with a high intensity in all P. granatum during the Growth Phenophase
studied cultivars. Total Chlorophyll
The highest photosynthesis process intensity Cultivar (mg/100g FW)
Hicaz 23.29
was determined at Hicaz, its value being 11.58
Kandahar 26.41
µmol/CO2/m2/s, followed by the Echen, 10.33
Nnikitski ranni 38.78
µmol/CO2/m2/s and Nikitski ranni, 9.71 Echen 28.14
µmol/CO2/m2/s (Table 2). Mollar 22.80
Shahvar 24.44
Table 2. The intensity of the Photosynthesis process in P. Dolce 22.09
granatum leaves during the Flowering Phenophase
Local selection 26.38
Light Temperature Photosynthesis
Cultivar intensity (°C) intensity
(µmol/m2/s) (µmol/CO2/m2/s) Regarding these 2 cultivars, there was a rele-
Hicaz 1402 18.8 11.58
vancy between the total leaf chlorophyll
Kandahar 1514 19.7 8.37
content and the intensity of the photosynthesis
Nnikitski 1415 18.2 9.71
ranni process. Therefore, the photosynthesis proce-
Echen 1425 19.5 10.33 sses of these cultivars showed the highest
Mollar 1505 18.5 9.03 intensity, given the highest content of
Shahvar 1498 18.2 5.19 chlorophyll (Tables 3, 1).
Dolce 1567 19.1 7.86 As for Hicaz, Kandahar, Mollar, Shahvar,
Local 1586 18.5 9.16 Dolce and the Local Selection cultivars, there
selection
was no interdependence between the total leaf
chlorophyll content and the intensity of the
The lowest intensity was recorded at Shahvar photosynthesis processes.
cultivar (5.19 µmol/CO2/m2/s).
There was recorded an increase of the photo- Total leaf Chlorophyll content of P.
synthesis process during flowering phenophase, granatum during the Flowering Phenophase
compared to growth phenophase, thus: at the During the flowering phenophase, it was noted
flowering phenophase moment, the Hicaz a different accumulation of chlorophyll pig-
cultivar intensified its photosynthesis process ments of the analyzed cultivars. The highest
95
values were shown at Kandahar (which The lowest transpiration intensity was recorded
flourished), Nikitski ranni (also flourished) and at Nikitski ranni, of 1.93 mmol/H2O/m2/s .
Shahvar, and the lowest were recorded at Analyzing the amount of water present in
Echen (Table 4). leaves and the intensity of transpiration
process, it was observed that the Echen cultivar
Table 4. Total leaf Chlorophyll content of correlates the considerable quantity of water
P. granatum during the Flowering Phenophase present in leaves with the high intensity of
Total Chlorophyll transpiration process (Table 5, 7).
Cultivar (mg/100g)
The studied cultivars carried out the transpi-
Hicaz 27.33
ration process with different values, albeit they
Kandahar 30.89
Nnikitski ranni 32.27
showed a similar water content in their leaves.
Echen 22.90 This occurrence is explained by the absorption
Mollar 28.89 of a distinct amount of light energy caused by
Shahvar 32.63 the difference in leaf size of each analyzed
Dolce 28.15 cultivar (the light intensity directly influences
Local selection 26.39
the transpiration process through its caloric
effect) (Burzo and Dobrescu, 2011; Dobrescu,
2007).
The Hicaz, Dolce and Mollar cultivars recorded
Table 5. The intensity of Transpiration process in P.
a higher chlorophyll content during flowering granatum leaves during the Growth Phenophase
phenophase, compared to growth phenophase,
with the remark that Dolce and Mollar reported Cultivar
Light
intensity
Temperature
(°C)
Transpiration
intensity
a similar increase rate of chlorophyll content. (µmol/m2/s) (mmol/H2O/m2/s)
These cultivars showed a connection between Hicaz 1245 18.0 3.30
the chlorophyll content and the intensity of the Kandahar 1189 18.1 2.45
photosynthesis process. At the Dolce cultivar, Nnikitski 1204 17.7 1.93
ranni
the photosynthesis process intensified with the Echen 1153 17.5 4.82
same rate that the chlorophyll content increased Mollar 1083 18.6 4.13
(1.25 times ) (Tables 4, 2). Shahvar 1073 19.0 4.58
It is worth mentioning that during the flowering Dolce 1126 17.7 2.27
phenophase, the chlorophyll content of Local Local 1119 17.8 3.64
selection cultivar did not change compared to selection
the growth phenophase and the photosynthesis
process was not influenced by this biochemical The intensity of the Transpiration process in
indicator. Therefore, these two physiological P. granatum leaves during the Flowering
parameters were not influenced by the pheno- Phenophase
phase at this cultivar. During the flowering phenophase, according to
specialty literature data, plant water
The intensity of the Transpiration process in requirements are higher and stimulate water
P. granatum leaves during the Growth absorption at the root level, which enhances the
Phenophase transpiration process (Burzo, 2016; Burzo et
Data analysis revealed an intense transpiration al., 2005; Dobrescu, 2007).
at Echen cultivar (4.82 mmol/H2O/m2/s), The transpiration process was carried out with a
followed by Shahvar (4.58 mmol/H2O/m2/s), distinct intensity from one cultivar to another:
and Mollar (4.13 mmol/H2O/m2/s) (Table 5). the highest transpiration intensities, with values
Referring to Echen cultivar, the intensity of between 5.15 and 5.56 mmol/H2O/m2/s were
transpiration process was: 1.41 times lower at recorded at Shahvar and Mollar cultivars,
Hicaz, 1.96 times lower at Kandahar, 2.49 followed by Echen and Local Selection
times lower at Nikitski ranni, 1.16 times lower cultivars of 4.34 and 4.90 mmol/H2O/m2/s.
at Mollar, 1.05 times lower at Shahvar, 2.12 The lowest values were registered at Nikitski
times lower at Dolce, and 1.32 times lower at ranni and Hicaz of 3.50 and 3.60
Local selection cultivar. mmol/H2O/m2/s (Table 6).
96
Table 6. The intensity of Transpiration process in P. Also, at Echen cultivar, it was recorded a
granatum leaves during the Flowering Phenophase high transpiration intensity, this process
Light Temperature Transpiration being conditional on the leaf water content
Cultivar intensity
(µmol/m2/s)
(°C) intensity
(mmol/H2O/m2/s)
(Tables 7, 5).
Hicaz 1402 18.8 3.60
The Dolce cultivar showed the lowest leaf
Kandahar 1514 19.7 4.12 water content, of 62.87%, compared to the
Nnikitski 1415 18.2 3.50 other analyzed cultivars. It was noted that, for
ranni this cultivar, the low leaf watter content
Echen 1425 19.5 4.34 diminished the intensity of photosynthesis and
Mollar 1505 18.5 5.56
transpiration processes (Tables 7, 1, 5).
Shahvar 1498 18.2 5.15
The rest of the studied P. granatum cultivars
Dolce 1567 19.1 3.92
showed an approximately equal leaf water
Local
selection
1586 18.5 4.90
content, with values between 64.8% and
65.8%.
The highest leaf dry matter content was regis-
The transpiration process was strongly influen-
tered at Dolce cultivar, of 37.13%, followed by
ced in its development by the phenophase,
Nikitski ranni and “Local Selection”.
stating that in all cultivars the transpiration
It was found that the Echen cultivar, whose
intensity increased during the flowering
leaves had the lowest dry matter content, of
phenophase, compared to growth phenophase,
31.82%, can not be explained because the
for example 1.12 times, respectively 1.34 times
intensity of the photosynthesis process was
at Shahvar and Mollar cultivars.
higher (it is known that photosynthesis process
High transpiration values during the flowering
leads to the accumulation of leaf dry matter)
phenophase can be explained due to both high
(Tables 7, 1).
light intensity and high temperature, external
Data obtained showed that the Kandahar,
factors that directly influence this process.
Mollar and Shahvar cultivars had a similar leaf
dry matter content, therefore this indicator does
Total leaf water and dry matter content of P. not allow the correlation with the
granatum during the Growth Phenophase photosynthesis process, process that presented
During growth phenophase, the highest water
fluctuations of values for each individual
content was determined at Echen cultivar, of
studied cultivar (Tables 7, 1). It is considered
68.18%, which was 1.05 times higher than the
that part of leaf dry matter resulted from
other studied cultivars (Table 7).
photosynthesis was translocated to growing
vegetative organs.
Table 7. Total leaf water and dry matter content of P. The accumulation of dry matter in plant organs
granatum during the Growth Phenophase
is mostly the result of the photosynthesis
Water content Dry matter efficiency. Also, the amount of dry matter in P.
Cultivar (%) content
(%) granatum leaves varies, depending on the
Hicaz 64.88 35.12 cultivar type, cultivar age, or the leaves
Kandahar 65.62 34.38 position on the branches.
Nnikitski ranni 64.40 35.60
Echen 68.18 31.82 Total leaf water and dry matter content of P.
Mollar 65.89 34.11 granatum during the Flowering Phenophase
Shahvar 65.71 34.29 During the flowering phenophase, the leaf
Dolce 62.87 37.13
water content of P. granatum cultivars showed
Local selection 64.55 35.45 a decreasement of approximately 1.04 times at
all studied cultivars compared to the growth
At Echen, the leaf water content had positively phenophase (Table 8).
influenced the process of photosynthesis which Also, it was found that the leaf water content
recorded the highest intensity (it is known that did not influence the transpiration process
the water represents a key matter for the which registered different intensities from one
photosynthesis process) (Tables 7, 1). cultivar to another (Tables 8, 6).
97
Table 8. Total leaf water and dry matter content of P. Table 9. The intensity of Respiration process in P.
granatum during the Flowering Phenophase granatum leaves during the Growth Phenophase
Water content Dry matter Light Temperature Respiration
Cultivar (%) content Cultivar intensity (°C) intensity
(%) (µmol/m2/s) (µmol/CO2/m2/s)
Hicaz 62.59 37.41 Hicaz 1245 18.0 - 2.77
Kandahar 64.26 35.74 Kandahar 1189 18.1 - 2.94
Nnikitski ranni 58.11 41.89 Nnikitski 1204 17.7 - 3.18
Echen 61.15 38.85 ranni
Mollar 63.27 36.73 Echen 1153 17.5 - 2.59
Shahvar 62.53 37.47 Mollar 1083 18.6 - 2.46
Dolce 61.60 38.40 Shahvar 1073 19.0 - 3.36

Dolce 1126 17.7 - 3.13
Local selection 63.18 36.82
Local 1119 17.8 - 3.06
selection
At Hicaz, Kandahar and Local Selection, the
leaf water content had values between 62.59% The lowest respiration intensity was registered
and 64.26%, lower than the ones registered in Mollar cultivar leaves, that also had the
during the growth phenophase. These values slightest biometry (based on macroscopic
are in interrelation with the high intensities of observations).
the transpiration process (some of the water The Shahvar cultivar had the highest
quantity present in leaves was eliminated) respiration process intensity.
(Table 8, 6).
The lowest leaf water content was determined The biochemical energy requirement for
at Nikitski ranni, of 58.11%. biosynthesis of organic substances involved in
During the flowering phenophase, the highest the growing process made the Shahvar cultivar
leaf dry matter content was recorded at Echen have the largest biometry (based on
cultivar, which was 1.22 times higher microscopic observations).
compared to growth phenophase. Also its
photosynthesis process intensified 1.25 times The intensity of the Respiration process in P.
(probably, the biosynthesized organic substance granatum leaves during the Flowering
following the photosynthesis process was Phenophase
present in the dry matter content) (Tables 8, 2). During the flowering phenophase there was a
Withal, the leaf dry matter content of Hicaz, difference in the floral evocation of the
Kandahar, Nikitski ranni, Mollar, Shahvar, analyzed cultivars, as follows: Hicaz,
Dolce, Local selection registered an increase- Kandahar, Nikitski ranni and Local selection
ment, compared to the growth phenophase, cultivar were characterized by the appearance
explained by a high photosynthesis process of the first flowers while the other cultivars
intensity (Tables 8, 2). Yet these cultivars’ leaf were not.
dry matter content was 1.15 times lower that It was revealed that the respiration process had
Echen’s. higher intensities in flowery cultivars compared
Therefore, this biochemical indicator can to the ones whose floral evocation did not
characterize and differentiate the analyzed happen (Table 10).
cultivars, influencing their metabolism and, the Analyzing the cultivars in terms of metabolism,
degree of acclimatization. namely the photosynthesis and respiration
values, representing the anabolic side,
The intensity of the Respiration process in P. respectively the catabolic side, it was found
granatum leaves during the Growth that the early flowery cultivars had a more
Phenophase intense metabolism compared to those of which
During the growth phenophase, the respiration the flowering process did not manifest during
process was carried out with different intensity the determination.
values at the analyzed cultivars: between -2.46
(Mollar) and -3.36 μmol/CO2/m2/s (Shahvar)
(Table 9).
98
Table 10. The intensity of Respiration process in P. Burzo I., 2016. Fiziologia procesului de înflorire.
granatum leaves during the Flowering Phenophase Elisavaros, București.
Burzo I., Delian E., Dobrescu A., Voican V., Bădulescu
Light Temperature Respiration L., 2004. Fiziologia plantelor de cultură, Vol. I,
Cultivar intensity (°C) intensity
(µmol/m2/s) (µmol/CO2/m2/s) Procesele fiziologice din plantele de cultură. Editura
Hicaz 1402 18.8 - 3.75
Ceres, București.
Burzo I., Delian E., Hoza D., 2005. Fiziologia plantelor
Kandahar 1514 19.7 - 4.85
de cultură, Vol. IV, Fiziologia pomilor, arbuştilor şi
Nnikitski 1415 18.2 - 4.50
ranni
plantelor ierboase fructifere. Elisavaros, București.
Burzo I., Dobrescu A., 2005. Fiziologia plantelor, Vol.
Echen 1425 19.5 - 3.55
VII, Fiziologia arbuștilor și plantelor lemnoase
Mollar 1505 18.5 - 2.88
spontane. Elisavaros, București.
Shahvar 1498 18.2 - 3.56 Dobrescu A., 2007. Fiziologia plantelor. Elisavaros,
Dolce 1567 19.1 - 3.61 București.
Local 1586 18.5 - 4.04 Hodgson R.W., 1917. The pomegranate. Bulletin of
selection California Agricultural Experiment Station. 76:163-
192.
Intrigliolo D.S.,Nicolas E., Bonet L., Ferrer P., Alarcón
CONCLUSIONS J.J., Bartual J., 2011. Water relations of field grown
Pomegranate trees (Punica granatum) under different
The Hicaz, Nikitski ranni, Echen, Mollar drip irrigation regimes. Agricultural Water
cultivars are the first ones that entered the Management, 98 (4): 691-696.
vegetative period, justifying a more intense Jamali B., Bonyanpour A.R., 2017. Evaluation of
adaptability potential of seven Iranian pomegranate
metabolic reactions during the research. cultivars in Southern Iran, Arsenjan region. Adv.
Only Hicaz, Nikitski ranni, Kandahar and Hort. Sci., 31 (2): 97-105.
Local Selection cultivars registered the floral Jurenka J., 2008. Therapeutic Applications of
evocation during the research, marking the Pomegranate (Punica granatum L.): A Review.
beginning of acclimatization process Alternative Medicine Review, 13 (2):128-144.
LaRue J.H., 1980. Growing pomegranates in California.
Regarding P. granatum species, the University of California Division of Agriculture
physiological processes and biochemical Science. Berkeley, Leaflet 2459.
parameters are directly influenced by the type Lichtenthaler H.K., Wellburn A.R., 1983.
of cultivar and by the phenophase. Determinations of total carotenoids and chlorophylls
a and b of leaf extracts in different solvents.
Biochem. Soc. Trans. 11, 591-592.
REFERENCES Parvizi H., Sepaskhah A.R., Ahmadi S.H., 2016.
Physiological and growth responses of pomegranate
Bădulescu L., 2016. Botanică și fiziologia plantelor. tree (Punica granatum L. cv. Rabab) under partial
Elisavaros, București. root zone drying and deficit irrigation regimes.
Burzo I., Dobrescu A., 2011. Stresul termohidric la Agricultural Water Management, 163 (1): 146-158.
plante - temperaturi ridicate și secetă. Editura Ceres,
București.
99
100
EFFECT OF DIFFERENT AMINO ACID FOLIAR FERTILIZERS

ON YIELD AND FRUIT QUALITY OF ʻREDIXʼ APPLE CULTIVAR
Alina Viorica ILIE1, 3, Cristina PETRIȘOR2, Dorel HOZA3

1
Research and Development Station for Tree Fruit Growing Băneasa, 4 Ion Ionescu de la Brad
Blvd, District 1, Bucharest, Romania
2
Research and Development Institute for Plant Protection, 8 Ion Ionescu de la Brad Blvd, District 1,
Bucharest, Romania
3
University of Agronomic Sciences and Veterinary Medicine of Bucharest, 59 Mărăşti Blvd,
District 1, Bucharest, Romania
Abstract
In fruit orchards the foliar application of nutrients is very important especially in certain periods when fruit trees
required important quantity of different compounds. The present study was aimed to determine the effect of foliar
application of amino acids on yield and fruit quality of ʻRedixʼ apple cultivar. The field experiments were carried in two
succesive seasons of 2016 and 2017, in a superintensive apple experimental orchard of USAMV Bucharest. The
fertilizers utilized in this experiment were Terra sorb complex, Terra sorb foliar, Naturamin, Hit AA in different
concentration. Fruit quality: average fruit size, weight, firmness, shape index (length/diameter) and also fruit chemical
features: dry matter, total soluble solids content of fruits and fruit acidity were recorded. Results indicates that all the
applied treatments were very effective in increase yield and fruit quality of ʻRedixʼ than control unfertilized.
Key words: firmness, weight, acidity, foliar application.
INTRODUCTION transportation of micro nutrients easier inside

the plant (Abo-Elmagd et al., 2015).
The production of apple using sustainable and Today, foliar spraying of aminoacids in most of
environmentally friendly agricultural practices the fruit orchards in the world has become
plays an essential role in determining their usual and its positive effects on fruit growth
market value and nutritional benefits. and quality characteristics are evident (Murtic
In recent years, different strategies have been et al., 2017; Arabloo et al., 2017; Molaie et al.,
applied to enhance the quality and productivity 2013; Fayek et al., 2011; Morales-Payan and
of apple without compromising quality Stall, 2003; Koksal et al., 1999).
standards (Tanou et al., 2017; Sala et al., 2014; The objectiv of this study were to test several
Jivan and Sala, 2013; Amiri et al., 2012). amino acids formulations by applied them
One of these is using fertilizers which can be during growth and development of fruit and
applied through the plant root suystem or determining the impact on quality and yield of
through leaves and each of these treatments has the ʻRedixʼ apple cultivar.
its advantages and disavantages (Murtic et al.,
2017; Dudu et al., 2015; Amiri et al., 2012; MATERIALS AND METHODS
Jafapour and Poursakhi, 2011).
Foliar fertilization has advantages of low The experiment was carried out at the USAMV
application rates, uniform distribution of Bucharest experimental orchard during the
fertilizer materials, easiest method of appli- period 2016-2017 in an superintensive orchard
cation and quick responses to applied nutrients planted in 2012 with ʻRedixʼ apple cultivar
(Abd El-Gleel Mosa et al., 2015). grafted on the M9 rootstock, at planting
Amino acids act as buffers for maintain distance of 3.5 x 1 m.
favorable pH value within the plant cell and In both years of investigation the following
also have a chelating effect on micro nutrients foliar fertilization treatments were applied to
when applied together making absorption and the plots:
101
Vo - Control (water only); RESULTS AND DISCUSSIONS
V1 - Terra sorb complex applied to leaves as a
0.2% solution; According to our results the foliar application
V2 - Terra sorb foliar applied to leaves as a of amino acids improved both quality and yield
0.2% solution; of ʻRedixʼ apple fruit.
V3 - Naturamin applied to leaves as a 0.2% The obtained results showed that the foliar
solution; application of amino acids gave great increases
V4 - Hit AA applied to leaves as a 0.2% in the yield, fruit firmness, average fruit
solution. weight, soluble solids content, and decreased
First treatment was applied at petal drop and the fruit acidity.
next treatments were done every two weeks. It can be noticed that foliar fertilizers
Each year, the chemical composition of the application does not significantly influence
fruits was analysed with specific methods in fruit diameter and fruit length in both seasons
three replications. Yield per tree (kg) was as compared to the control.
measured on five trees on three replications. A The fruit weight of apple increased signify-
sample of randomly picked 15 fruits per cantly from 173.26 g in the control variant to
cultivar was harvested at commercial maturity 188.80g in Terra sorb foliar variant (V2) and
for determining of quality traits. 190.86 g in Naturamin variant (V3) and also
Dry matter of fruits was determined with a 225 g in Hit AA (V4) in 2016 year (Table 1).
gravimetric method through drying an aliquot The similar results were obtained by Arabloo et
~5 g of fruit tissue at 105°C to constant weight. al., 2017 who have found a significant increase
Titratable acidity (TA) was determined by in fruit weight for ʻGolden Deliciousʼ and
titration of an extract of fruit homogenate with ʻGranny Smithʼ cultivars fertilized with amino
0.1 N NaOH to the end point of pH 8.1 and acids.
expressed as malic acid percent. The average fruit weight was higher in 2017
Soluble solids (SS) expressed as °Brix were compared to 2016.
measured in juice pressed from the whole fruit The all foliar application also resulted in a
sample using a digital refractometer (model significant increase in yield of apple fruit as
PR-101, ATAGO, Tokyo, Japan) at 21°C. compared to the control. Kamiab et al. (2015)
Fruit firmness (kg/cm2) was measured using a reported that foliar spraying of amino acids
penetrometer (FT-327) with a 11-mm diameter increased the quantitative and qualitative
probe from three different areas (top, middle characteristics of pistachio.
and bottom) of the whole fruit.
Table 1. Influence of foliar fertilizer on fruit size, fruit weight and fruit yield
of ʻRedixʼ cultivar during 2016-2017
Treatment Fruit weight Fruit diameter Fruit length Fruit yield/tree Yield
(g) (mm) (mm) (kg) (t/ha)
2016 2017 2016 2017 2016 2017 2016 2017 2016 2017
V0 - water 173.26 183.06 77.00 70.60 9.03 8.71 10.57 12.27 30.20 35.06
V1 - Terra sorb 175.21 186.75 75.00 71.05 10.51 9.52 20.32 18.30 58.05 52.28
complex
V2 - Terra sorb 188.80 198.28 77.50 73.70 10.33 9.82 24.36 16.66 69.60 47.60
foliar
V3 - Naturamin 190.86 192.21 79.00 74.55 9.13 10.20 20.07 21.33 57.34 60.94
V4 - Hit AA 225.12 197.99 81.20 71.80 9.06 10.68 21.84 30.69 62.40 87.68
Data presented in table 2 show that in all foliar determined in variant V4 (16.35 g%) followed
treatments (V1-V4), dry matter content was by the variant V3 (16.32 g%) and V2 (16.07
higher in comparison to control variant (V0) g%) compared to the control variant V0 (14.91
both in 2016 as weel as in 2017. The highest g%). The values of this compound are slightly
dry matter content in apple fruit was low in 2017 compared with 2016.
102
Table 2. Influence of foliar fertilizer on fruit quality parameters of ʻRedixʼ cultivar in 2016-2017
Treatment Dry matter Firmness Soluble solids Titratable acidity SS/TA ratio
(g%) kg/cm2 °
Brix (malic acid %)
2016 2017 2016 2017 2016 2017 2016 2017 2016 2017
V0-water 14.91 14.65 9.03 8.71 10.62 10.2 0.96 0.85 11.06 12
V1 - Terra sorb 15.60 14.98 10.51 9.52 12.04 11.03 0.85 0.82 14.16 13.45
complex
V2 - Terra sorb 16.07 15.08 10.33 9.82 11.10 10.65 0.91 0.84 12.20 12.68
foliar
V3 - Naturamin 16.32 15.11 9.13 10.20 12.78 11.76 0.88 0.80 14.52 14.70
V4 - Hit AA 16.35 15.68 9.06 10.68 13.80 12.20 0.81 0.79 17.04 15.44
Foliar application of V1 and V2 foliar Application of Naturamin and Hit AA

fertilizers resulted in a significant increase in fertilizers had the highest positive effect to
apple fruit firmness. Firmness of the fruit to improve the percentages of yield and average
which the Naturamin (9.13 kg/cm2) and Hit AA fruit weight. Also, it increased dry matter
(9.06 kg/cm2) fertilizers has been applied was content of apple in both seasons, as compared
very close to the value of the control (9.03 to the control treatment.
kg/cm2) in 2016 year. Similar results for fruit All fertilizers used decreased the percentage of
firmness were obtained by Arabloo et al., 2017 acidity, but differences were not significant
on ʻGolden Deliciousʼ and ʻGranny Smithʼ among treatments for each year studied.
apple cultivars. Milosevic and Milosevic, 2015
reported that spraying with foliar fertilizer REFERENCES
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cultivar . Abd El- Gleel Mosa W.F.A., Abd EL-Megee N.A., Paszt
L.S., 2015.The effect of the foliar application of
The TA content of apple fruit was not potassium, calcium, boron and humic acid on
significantly affected by the application of vegetative growth, fruit set, leaf mineral, yield and
foliar fertilizers. However application of fruit quality of ʻAnnaʼ apple trees. American Journal
Naturamin and Hit AA fertilizers decreased of Experimental Agriculture. 8 (4): 224-234.
titratable acidity percent. Abo-Elmagd N.A., Nasr M.M., Ahmed N.A., 2015.
Enhancing fruit quality and storability of ʻAnnaʼ
From the results obtained, the all foliar apple cultivar by using amino acids, ethylene and
application of amino acids improved TSS and some nutrients middle east. Journal of Agriculture
TSS/ acid ratio and decreased acidity per- Research. 4 (4): 802-812.
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in the two seasons studied. The results of our foliar and ground fertilization on yield, fruit quality,
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experiment confirm data obtained by Abo- Plant Nut. 31 (3): 515-525.
Elmagd et al., 2015, that found a positive effect Arabloo M., Taheri M., Yazdani H., Shahmoradi M.,
of amino acids on the soluble solids content of 2017. Effect of foliar application of amino acid and
apple fruit. However results obtained by Malik calcium chelate on some quality and quantity of
and Singh, 2006 showed that total soluble Golden Delicious and ʻGranny Smithʼ, Trakia Journal
of Sciences, 1: 14-19.
solids content of fruit was reduced by the Dudu G., Cimpeanu S.M., Jerca O.I., 2015. The
aqueous solutions of amino acids when sprayed influence of biofertilization upon production level of
on mango, cv. ʻKensington pride" like foliar some harvested apple cultures (Vf) in an intensive
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foliar spray on fruit quality was also supported Papers, Series B, Horticulture, LIX: 43-46.
Fayek M.A., Yehia T.A., El-Fakhrany E.M.M, Farag
by other authors (Fayek et al., 2011; Khan et A.M., 2011. Effect of ringing and amino acids
al., 2012). application on improving fruiting of ʻLe Conteʼ pear
trees. Journal of Horticultural Sci. & Ornamental
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Khan A.S., Ahmad B., Jaskani M.J., Ahmad R., Malik
A.U., 2012. Foliar application of mixture of amino
The foliar application of amino acids had a acids and seaweed (Ascophylum nodosum) extract
positive effect to improve the yield and fruit improve growth and physicochemical properties of
quality of ʻRedixʼ apple trees. grapes. Int J. of Agric & Biol, 14 (3): 383-388
103
Kamiab F., Heidari M., Zamani E., 2015. Evaluation the on photosynthesis and yield of two commercial
effects of foliar treatments of polyamines and some cultivars in pistachio orchards of Kerman province in
organic acids on quantitative and qualitative traits in Iran. Inter. J. Agri. Crop Sci, 23: 2827-2830.
some pistachio cultivars. Journal of Nuts, 6 (2): 131- Morales Payan J.P., Stall W.M., 2003. Papaya (Carica
142. papaya) response to foliar treatments with organic
Koksal A.I., Dumanoglu H., Gunes N.T., 1999. The complexes of peptides and amino acids. Proc. Fla.
effects of different amino acid chelate foliar State Hortic. Soc, 116: 30-32.
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Fe, Zn, Cu, Mn content of leaves in ʻWilliamsʼ pear quality and calcium content as affected by fertilizer
cultivar (Pyrus communis L.). Turkish Journal of treatment. Pol. J. Environ. Stud., 26 (5): 2107-2111.
Agriculture and Forestry, International Conference on Sala F., Iordănescu O., Dobrei A., Jivan C., 2014.
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Jivan C., Sala F., 2013. Variation of foliar area in apple supplement in achieving apple production. Journal of
in response to nutrition guidance through foliar Horticulture, Forestry and Biotechnology, 18 (2):
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Malik A.U., Singh Z., 2006. Improved fruit retention, Balakrishnamurthy G., 1998. Effect of micronutrients
yield and fruit quality in mango with exogenous on yield and quality of mandarin orange (Citrus
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167-174. 131.
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Molaie H., Panahi B., Tajabadipour A., 2013. The effect
of foliar application of some amino acid compounds
104
RESEARCHES ON GENETIC RESISTANCE TO APRICOT VARIETY

’BERGERON’ TO FROST FROM WINTER
Constantin PĂUN

Abstract
The research was conducted in a orchard of apricot aged 7 years, located at 50 km away from Bucharest. Trees planted
at a distance of 4x4m were studied during the vegetation season paying particular attention to resistance to low
temperature. In the winter of 2013, temperatures in winter decreased to -20 degree C and produced losses of flower
buds. In the spring of 2014 the ‘Bergeron’ variety bloomed well, 100% of flower buds being resistant to negative
temperatures from winter. In 2015 was a warm winter but the temperature decreased in January to -29 degree C and
the trees suffered a thermal shock. Following, resistance to frost decreased and the percentage of flower buds affected
was 98.2%. Production losses were very high and vegetative buds were not affected by frost and the trees were properly
vegetated grew normally. In January of 2016 were recorded temperatures of -25 degrees C associated with very cold
wind, the flower buds were affected in 100%. In 2017 ‘Bergeron’ variety was not affected.
Key words: flower buds, temperatures, thermal shock.
The apricot is a species appreciated for fruit Research and observations were made in an
quality and is grown successfully in areas with orchard set up in 2010, located at a distance of
high temperatures (Cociu V. et al., 2000; 50 km N, compared to Bucharest. The studies
Sestraș R. et al., 2004). In Romania, grows in began in 2013, when the orchard was in the 3rd
the S part of the country and in the Plain of V. year of vegetation, and the trees began to fruit
In Romania, the apricot, do not make the fruits and continued until 2017. The biological
every year because of the low temperatures that material studied was the ‘Bergeron’ apricot
affect the flowers (Braniște N. et al., 2007). variety produced in Romania, grafted on the
The ‘Bergeron’ is a new variety introduced in Myrobolan 29 C rootstock. The trees were
crop and has not been studied for resistance to planted at a distance of 4 m between the rows
low winter temperatures and temperatures and 4 m between the plants per row and were
during the flowering period (Ghena N., led as a vessel. The fructification cuts were
Braniște N., 2003). made in August, after harvesting the fruit. Soil
We made a trial with ‘Bergeron’ variety to has been kept clean by weeds by manual and
study resistance to winter cold, fructification mechanical work. Were studied 50 trees in 5
and fruit quality. rehearsals. To record the temperatures during
As a result of the researches we were able to winter and during the vegetation period we
establish the resistance limit at low used our own thermometers and meteorological
temperatures and I observed that the ‘Bergeron’ data from the nearby weather station. To
variety behaves differently from one year to determine the percentage of buds affected by
another depending on the evolution of the low winter temperatures, we made observations
temperatures. In cold winters associated with in the laboratory where we analyzed at
wind, production losses are very high, up to microscope the fruit buds and vegetative buds
100% (Păun C., 2017). and in the spring we made observations on the
Researchers are concerned about obtaining field. Observations were made in November
apricot varieties that start late in vegetative and (before frost), in February, at the beginning of
resist winter frost. the vegetative period (March) and during the
105
vegetative period. After the flowering we Table 3. Behavior of the ‘Bergeron’ variety
determined the percentage of flowers formed to in the winter 2016
determine the influence of winter cold on fruit % %
buds. During the vegetation period, we made Variety Rehearsal Nr. buds viable affected
analyzed fruit buds
observations on the percentage of fruit formed, buds
the length of the shoots, the production of the R1 620 0 100
fruit obtained and the qualities of the fruits. The R2 514 0 100
shoots were measured at the end of July, after ‘Bergeron’ R3 387 0 100
stopped of growth. The fruits were weighed in R4 427 0 100
the laboratory and analyzed for determinate the R5 489 0 100
Average 487.8 0 100
average fruit weight, colour, pulp weight and
kernel weight. Table 4. Behavior of the ‘Bergeron’ variety
in the winter 2017
RESULTS AND DISCUSSIONS % %
Variety Rehearsal Nr. buds viable affected
In the 3rd year of vegetation, the ‘Bergeron’ analyzed fruit buds
variety went through the winter without losing buds
the fruit buds. This year, the first fruit R1 382 98.7 0.7
R2 417 98.4 0.6
production was obtained. ‘Bergeron’ R3 354 99.7 0.3
Research into the resistance of fruit buds to R4 472 99.4 0.6
winter cold began in the winter of 2014 when R5 395 99.3 0.7
the trees were 4 years old. Average 404 99.1 0.5
The year 2014 was a normal year from the
thermal point of view and the trees resisted Table 5. Analysis of the influence of low temperatures
very well during the winter. on the percentage of fruit formed
In spring, the trees flourished normally without Variety 2014 2015 2017
the loss of fruit buds. Rehearsal % fruit % fruit % fruit
formed formed formed
Fruit buds resisted 100% and fruit production R1 31 58 27
was normal (Table 1). R2 28 60 31
‘Bergeron’ R3 38 62 25
Table 1. Behavior of the ‘Bergeron’ variety R4 37 58 28
in the winter 2014 R5 35 55 24
% % Average 33.8 58.6 27
Variety Rehearsal Nr. buds viable affected
analyzed fruit buds
buds
In 2014 and 2017, winter temperatures
R1 380 100 0 decreased to -25 ° C, but the temperature drop
‘Bergeron’ R2 300 100 0 was gradual, and the buds were resistance and
R3 420 100 0 was achieved a normal production. In 2015 the
R4 200 100 0 heat shock destroyed the buds, and in 2016 the
R5 470 100 0 very cold wind from the winter period
Average 354 100 0
amplified the cold and destroyed all the fruit
buds (Table 2 and Table 3). Observations
Table 2. Behavior of the ‘Bergeron’ variety
in the winter 2015 regarding the genetic resistance of flowers at
low temperatures during bloom showed that the
% %
Variety Rehearsal Nr. buds viable affected ‘Bergeron’ variety very well tolerates low
analyzed fruit buds temperatures. In years with high production,
buds when winter frost did not destroy fruit buds, the
R1 500 1.4 98.6 flowers were not affected by low temperatures
R2 347 2.7 97.3 during the flowering period. In 2014 and 2017
‘Bergeron’ R3 289 1.6 98.4
during the bloom when the flowers were
R4 320 1.5 98.5
R5 415 1.8 98.2 opened, during the night were temperatures of
Average 374,2 1.8 98.2 -3 ... -4 degree C, and the apricot flowers were
106
not affected (Table 4). The results regarding the (Table 7). At the end of the growing season
influence of low temperatures on the percen- (July), the shoots were measured in length and
tage of fruits formed are presented in table 5. the number of vegetative and fruit buds was
During the vegetation period observations were determined on each shoots for estimate the
made on the length of the shoots, number of production of the following year.
shoots/tree, number of vegetative buds/shoots The correlation between fruit weight and pulp
and number of fruits buds/shoot (Table 6). weight is presented in figure 1, and the
The fruits were analyzed morphologically and correlation between fruit weight and pulp
was determined the production on the tree weight is presented in figure 2.
Table 6. Observations on the growth of shoots

Variety Year The length of shoots Nr. of shoots/tree Nr. of vegetative Nr. of fruit
(cm) buds/shoots buds /shoot
2014 62 120 58 121
2015 58 134 49 102
‘Bergeron’ 2016 50 119 47 94
2017 45 143 38 84
Average 43 102.4 38.4 80.2
Table 7. Observations on characteristics fruits and productivity
Average fruit weight Weight of pulp Weight of Fruit production

Variety Year
(g) (g) kernel (g) (kg/tree)
2014 86 80.2 5.8 14
2015 90 84.3 5.7 2
‘Bergeron’ 2016 0 0 0 0
2017 92 85.9 6.1 21
Average 89.3 83.4 5.8 12.3
g pulp (g)
88
86 R² = 0,9967
84
82
80
78
85 86 87 88 89 90 91 92 93
Weight of fruit ( g)
Figure 1. The correlation between fruit weight and pulp weight
g. kernel (g)
6,2
6 R² = 0,3324
5,8
5,6
85 86 87 88 89 90 91 92 93
Weight of fruit ( g)
Figure 2. The correlation between fruit weight and pulp weight
107
CONCLUSIONS normally, being resistant to low temperatures
during the flowering period
‘Bergeron’ variety is a relatively resistant
variety to low winter temperatures and REFERENCES
resistance depends on how low the temperature
is. Braniște N. et al., 2007. Soiuri de pomi, arbusti fructiferi
For protect the ‘Bergeron’ variety to low si capsuni create in Romania. Editura Paralela 45.
Cociu V. et al., 2000. Progrese in ameliorarea plantelor
temperature from winter, the orchard must be horticole. Editura Ceres, vol. I, Pomicultură, 215 pag.
established in areas where the wind does not Ghena N., Braniște N., 2003. Cultura speciala a pomilor.
blow. Editura Matrix Rom, București, 399 pag.
Under thermal shock, fruit buds can be affected Păun C., 2017. Pomicultura in imagini. Editura La
to 98.2%, and cold wind causes 100% fruit bud Sanziene.
Sestraș R. et al., 2004. Ameliorarea speciilor horticole.
losses. Editura Academic Pres, Cluj-Napoca.
If the fruit buds are not affected during the
winter, the ‘Bergeron’ variety fructify
108
STUDY OF THE CYTOLOGICAL CHARACTERISTICS

AND GERMINATION POTENTIAL OF SOME PEACH CULTIVARS
FROM RSFG CONSTANŢA
Corina GAVĂȚ1, Elena DOROFTEI2, Liliana MIRON2,

Vlăduț Alexandru OPRIȚĂ1, Georgeta BELENIUC2
1
Research Station for Fruit Growing Constanţa, No. 25 Pepinierei Street,
907300, Valu lui Traian, Constanţa, Romania
2
Ovidius University Constanţa, 124 Mamaia Blvd, Constanţa, Romania

Abstract
The paper shows the results of the genetic potential of some peach cultivars (Prunus persica L.) used as genitors in the
breeding process. A detailed morphological and structural description of pollen grains of four peach cultivars ‘Raluca’,
‘Florin’, ‘Filip’ and ‘Monica’ was done. The pollen was collected from peach flowers in April from the demostrative
plots at Research Station for Fruit Growing Constanta (RSFG Constanta). The viability and pollinated tube cultures
were made in order to determine the fertility and germination capacity of the analyzed biological material. The one
with the highest viability of the pollen grains was recorded at ‘Raluca’. ‘Florin’ cv. has been shown the highest
germination capacity on a 15% sucrose nutrient medium after a 2 hour sprouting time (89.6%), followed by a 5%
sucrose nutrient medium with after 2 hours of germination time (88.9%).
Key words: Prunus persica L., pollen, sucrose.
The peach is one of the most appreciated In order to analyze the viability of the pollen,
species that is very very well adapted to the were used substances that bind H2-catalysed
southeastern part of the country. dehydrazes resulting a new colored substance
The studied peach cultivars were created to (formazam) that distinguished the viable pollen
RSFG Constanta and belong to the fresh from the unviable one. The mature anthers
consumption type. strike on a lamella in a drop of 0.5% carmin
Pollen sterility is a character that is finding in acetic solution; 3 - 5 preparations are microsco-
few cultivars of peach, being a recessive and pic analysis, respectively 5 microscopic fields
monogamy character (Scott and Weinberger, of each preparation; microscopic fields include
quoteds by Cociu V., 1981). red pollen granules because viable granules are
Reached by stigma, due to wind, insects, birds, colored in red while unviable pollen retains its
water and humans, the pollen grains are soaked original yellow colored and there are wrinkled
with water from the stigmatic liquid, it is swells (Andrei and Paraschivoiu, 2003; Doroftei et al.,
and germinates: the exine cracks and intine 2008). The statistical interpretation was based
exits out like a pollinic tube through the on the number of germinated and non-
germination pore. germinated pollen grains from a total of 1000
The pollen is retained by the stigma and pollen grains from the analyzed microscopic
absorbs the water from the adhesive mixture fields in 3 repetitions, in order to determine the
that is produced by it; it is moisturizes and then germination capacity of pollen grains.
germinates. For microscopic analysis, observations were
In general, pollen moisturizing lasts from few made and images were taken on a FLUO 2
seconds to few minutes and germination begins research microscope provided with a BEL
after 45-210 minutes, both phenomena PHOTONICS DV-1300 video camera.
depending on the species (Andrei, 1978; Determination of pollen germination
Şerbănescu-Jitariu and Toma, 1980). capacity was achieved by collecting pollen of
109
peach cultivars at the time of full flowering: put average dimensions ranging from 19-30 μm
5%, 10% and 15% sucrose solutions on one (Table 1).
blade, sprinkle mature pollen on each blade and The pollen membrane has a complex compo-
analyzed at a microscope at 30 minute intervals sition that differs from the outside consisting of
(up to two hours); the coloring with a 0.5% ectexin and endexin.
carmin acetic solution for 2-3 minutes, then Exina is covered with tubers or beaks of
covering with the lamella. Three to five prepa- different shapes and sizes, with positive ele-
rations are analyzed for each experimental ments prevailing, the surface of which is
variant, 5 microscopic fields of each prepa- praying, with ornamental elements larger than
ration are numbered, and germinated and non- one micrometer, elongated and distributed
germinated pollen granules are included in the irregularly.
microscopic fields (Andrei and Paraschivoiu, In the field of microscopy they appear as elon-
2003; Doroftei et al., 2008). gated, irregularly spaced ridges (Table 1,
In order to determine the germination capacity Figure 1).
of pollen grains the statistical interpretation Apertures or germinating apertures include
was based on the number of germinated and colps and pores. Colps have different shapes
non-germinated pollen grains from a total of and can form with the granule (with regular
1000 pollen grains from the analyzed micros- margins) or later than the granule (with
copic fields in 3 repetitions. For microscopic irregular margins).
analysis, observations were made and images On the analyzed microscopic preparations we
were taken on a FLUO 2 research microscope usually observed 3 colts, but their number can
provided with a BEL PHOTONICS DV-1300 be between 2 and 4.
video camera. The pores are cone-shaped acamerate. The
main pollen morphological types observed at
RESULTS AND DISCUSSIONS the microscope were: bicolpat, polarized or
tricolpic pollen, tricolor (Table 1).
Morphological an structural characteristics
of pollen grains to peaches cultivars studied Table 1. The characteristics of the analized pollen
The cross sections in the antennas at different from four peach cv.
stages point to the existence of a homogeneous Average Elements of
mass of meristematic cells (at the beginning); Cultivars
size
Shape
the exine
Apertures
differentiation of the array of hypodermic cell ʻRalucaʼ 28 μm triangular, rugulată tricolpat

spheroidal triporat
strings (at a more advanced stage); repeated ʻFlorinʼ 30 μm triangular, rugulată tricolpat
division of the anther's cells and differentiation triporat
ʻFilipʼ 26 μm triangular, rugulată tricolpat
of the primary parietal layer as well as the spheroidal triporat
primary sporogen layer. monocolpat
monoporat
The parietal layers that appear a little bit later ʻMonicaʼ 23 μm triangular, rugulată tricolpat
are: epidermis, endothelium, several medium oval triporat
bicolpat
layers and the inner most layer called biporat
wallpaper, whose physiological importance is
special because it serves to nourish spores.
The sporogenic tissue has as orgin primary
sporogenic cells. Some of the sporogen cells
degenerated frequently and are absorbed by the
rest of the cells. The resulted microspors from
the microsporus parent cell divisions are
disposed in tetrades that can be of different
types.
The distinct elements of the pollen grains are
the polar axis, the equatorial axis, and the
poles. Pollen granules analyzed have different Figure 1. Morphological aspect of pollen grains in
shapes: spherical, oval or triangular, with Prunus persica L. ʻMonicaʼ (1000x). There are exine,
intine, 3 germinating pores and the surface of the exine
110
The viability of the pollen it can be seen as time (88.9%). This cultivar proved to be fertile,
one of the most fertile peach from the cultivars although viability was the smallest of the four
analyzed, with the highest viability is 'Raluca' analyzed varieties (74.2%) (Table 3).
and low fertile with minimal viability 'Florin' At ‘Raluca’ cultivars, the highest germination
(Table 2). capacity was observed on a 5% sucrose nutrient
Table 2. The viability of the pollen for the analized medium after a 2 hours (85.9%) germination
cultivars time (Figure 3) and the lowest on a 10%
Cultivars The total The The Viability
sucrose nutrient medium after a germination
number of number of number of % time of 30 minutes (22%) (Table 3).
the analized unviable viable
pollen pollen pollen
At the ‘Filip’ cultivars, the highest germination
grains grains grains capacity was observed on a 5% sucrose nutrient
ʻRalucaʼ 1000 125 875 87.5
medium after a 2 hours germination time
ʻFlorinʼ 1000 258 742 74.2
ʻFilipʼ 1000 189 811 81.1
(72.4%) (Figure 4) and the lowest on a 15%
ʻMonicaʼ 1000 176 824 82.4 sucrose nutrient medium after a germination
time of 30 minutes (42%).
Germination capacity The comparative ʻMonica'sʼ cultivar highest germination
analysis of the four peach cultivars showed that capacity was observed on a 10% sucrose
the highest germination capacity was observed nutrient medium after a 2 hours germination
to the ‘Florin’ cultivar on a 15% sucrose nu- time (61.2%), and the lowest on a 5% sucrose
trient medium after a 2 hours germination time nutrient medium after a germination time of 30
(89.6%) (Figure 2), followed by 5% sucrose minutes (24.3%) (Figure 5).
nutrient medium after 2 hours of germination
Table 3. The germination capacity of analyzed peach cultivars
Non-
Total number of Germinated Germination
Cultivars Treatment germinated
pollen grains pollen grains capacity %
pollen grains
ʻRalucaʼ Distillated water (Control) 1000 0 1000 0
Sucrose 5%, 30 minutes 1000 556 444 55.6
Sucrose 5%, 1 hour 1000 668 332 66.8
Sucrose 5%, 2 hours 1000 859 111 85.9
Sucrose 10%, 30 minutes 1000 220 780 22.0
Sucrose 10% 1 hour 1000 276 724 27.6
Sucrose 10% 2 hours 1000 283 717 28.3
Sucrose 15%, 30 minutes 1000 478 522 47.8
Sucrose 15%, 1 hour 1000 544 456 54.4
Sucrose 15%, 2 hours 1000 604 396 60.4
ʻFlorinʼ Distillated water (Control) 1000 0 1000 0
Sucrose 5%, 30 minutes 1000 678 322 67.8
Sucrose 5%, 1 hour 1000 790 210 79.0
Sucrose 5%, 2 hours 1000 889 111 88.9
Sucrose 10%, 30 minutes 1000 584 416 58.4
Sucrose 10% 1 hour 1000 667 333 66.7
Sucrose 10% 2 hours 1000 834 166 83.4
Sucrose 15%, 30 minutes 1000 632 368 63.2
Sucrose 15%, 1 hour 1000 821 179 82.1
Sucrose 15%, 2 hours 1000 896 104 89.6
ʻFilipʼ Distillated water (Control) 1000 0 1000 0
Sucrose 5%, 30 minutes 1000 482 518 48.2
Sucrose 5%, 1 hour 1000 663 327 66.3
Sucrose 5%, 2 hours 1000 724 276 72.4
Sucrose 10%, 30 minutes 1000 355 645 35.5
Sucrose 10% 1 hour 1000 507 493 50.7
Sucrose 10% 2 hours 1000 654 346 65,4
Sucrose 15%, 30 minutes 1000 420 580 42.0
Sucrose 15%, 1 hour 1000 506 494 50.6
Sucrose 15%, 2 hours 1000 591 409 59.1
ʻMonicaʼ Distillated water (Control) 1000 0 1000 0
Sucrose 5%, 30 minutes 1000 243 757 24.3
111
Non-
Total number of Germinated Germination
Cultivars Treatment germinated
pollen grains pollen grains capacity %
pollen grains
Sucrose 5%, 1 hour 1000 305 695 30.5
Sucrose 5%, 2 hours 1000 347 653 34.7
Sucrose 10%, 30 minutes 1000 462 538 46.2
Sucrose 10% 1 hour 1000 506 494 50.6
Sucrose 10% 2 hours 1000 612 388 61.2
Sucrose 15%, 30 minutes 1000 269 731 26.9
Sucrose 15%, 1 hour 1000 318 682 31.8
Sucrose 15%, 2 hours 1000 364 636 36.4
Figure 2. Pollen grains of Prunus persica ‘Florin’ on sucrose 15%, 2 hours (400x)
Figure 3. Pollen grains of Prunus persica ‘Raluca’ on sucrose 5%, 2 hours (400x)
Figure 4. Pollen grains of Prunus persica ‘Filip’ on Figure 5. Pollen grains of Prunus persica ‘Filip’ on
sucrose 5%, 2 hours (400x) sucrose 5%, 30 minutes (100x)
112
CONCLUSIONS sprouting time (89.6%) followed by the 5%
sucrose nutrition medium after 2 hours of
The analyzed pollen granules have different germination time (88.9%).
shapes: spherical, oval or triangular, with
average dimensions ranging from 19-30 μm. ACKNOWLEDGEMENTS
Exina is covered with tubers or beaks of
different shapes and sizes with positive This research work was carried out with the
elements predominantly, the surface is rugulate, support of Ministry of Agriculture and Rural
with ornamental elements larger than one Development, and was financed from Project
micrometer, elongated and distributed ADER 3.2.2.
irregularly. Three colps and three pores were
usually observed on the analyzed microscopic REFERENCES
preparations, but their number may range from
2 to 4. The pores are conical. Andrei M., 1978. Anatomia plantelor. Editura Didactică
From all peach varieties analyzed the one with şi Pedagogică, Bucuresti, 321-326, 339-343.
Andrei M., Anghel I., Popescu I., Stoica E., 1981.
the highest viability is ‘Raluca’ and the least Lucrări practice de biologie vegetală. Editura
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The comparative analysis of the five peach and Andrei M., Paraschivoiu R.M., 2003. Microtehnica
nectarine varieties showed that the highest botanica. Editura Niculescu, Bucureşti.
germination capacity was observed in the Cociu V. si colab, 1981. Cultura piersicului. Editura
Ceres, Bucuresti.
ʻFlorinʼ variety on a 15% sucrose nutrient Doroftei E., Rotaru-Stăncic M., Dobrinas S., Mandalopol
medium after a 2 hours sprouting time (89.6%) D., 2008. Relationships between metals extent and
followed by the medium 5% sucrose nutrition cytological characteristics at Prunus persica L.
after 2 hours of germination time (88.9%). pollen. Environmental Engineering and Management
The comparative analysis of the four peach Journal, Vol.7, No.2, 97-101.
Şerbănescu-Jitariu G., Toma C., 1980. Morfologia şi
cultivars showed that the highest germination anatomia plantelor, E.D.P., Bucureşti.
capacity was observed in the ʻFlorinʼ cv. on a
15% sucrose nutrient medium after a 2 hours
113
114
BEHAVIOR OF SOME APRICOT CULTIVARS GRAFTED

ON NEW VEGETATIVE ROOTSTOCKS
Vlăduț Alexandru OPRIȚĂ, Corina GAVĂȚ
Research Station for Fruit Growing Constanţa, No. 25, Pepinierei Street,
907300, Valu lui Traian, Constanţa, Romania
Abstract
In the Dobrogea area an important component in obtaining orchards with notable economic performances is the
finding of the best variety-rootstock combinations. The different physical and chemical structure and composition of
soil in Dobrogea obliges to find scientific solutions that will come to the future farmers. At RSFG Constanta in the
period 2013-2018, the compatibility and development of a large apricot species grafted on different rootstocks
originating in the Mediterranean area was assessed. Thus, the ʻAmiralʼ apricot variety was grafted on the following
rootstocks: Adesoto, Myrobolan 29 C, Weiwa, Ishtara. As a result of the measurements and the analyzed data, it was
noticed that the Ishtara rootstock shows an increased compatibility and, once again, imparts a medium vigour, which
means that by introducing the multiplication of this rootstock we have high-density orchards, so the fruit growing in the
Dobrogea area a model to be followed for other apricot-favorable areas.
Key words: Prunus armeniaca (L.), compatibility, orchard systems.
INTRODUCTION annual precipitation amount is 400 mm, out of

which during the growing season (April 1 to
The used rootstocks for apricot are originated September 30), 240.7 mm. The lowest winter
from six species of Prunus genus: P. temperatures below -20°C are not very often: 1
armeniaca L., P. cerasifera Ehrh., P. domestica out of 10-15 years and so are the spring frosts
L., P. insititia Jussl., P. persica Stock., P. susceptible to cause apricot yield damage.
amygdalus L. (Cociu et al., 1993). All over the The climatic water deficit reaches as much as
world the apricot rootstocks are used according 400 mm/year, so irrigation application is
to the pedoclimatic conditions, affinity, needed for apricot. The zonal soil type is a
cropping systems, etc. Among the most calcaro-calcic chernozem formed on loess, with
frequently used are small apricot, franc apricot, loam texture and a proper capacity of water
Manicot (GF 1236), cherry plum, myrobolan preserving, holding and circulation. The humus
plum, franc plum (Bălan et al., 2008). content ranges between 2.5 and 4%; pH of the
soil is poor alkaline (7.0-8.1).
MATERIALS AND METHODS Observations and determinations were
observed in Field I and II within the RSFG fruit
The study was carried out in apricot tree nursery. In the third year (2017) fifty fruit
demonstrative lots at Research Station for Fruit trees of ʻAmiralʼ cv. grafted on each mentioned
Growing Constanta, located in south-eastern vegetative rootstocks were planted in an
Romania, near the Black Sea. experimental field. The planting scheme was
The site is located at 44°10’ Northern latitude chosen 4/4 m, the orientation of the rows was
and 28°29’ Eastern longitude, and 70 m above from North to South and the shape of the crown
sea level. Climate is continental with warm and was improved vase.
droughty summers, frequent dry winds all the
year round and temperate winter generally RESULTS AND DISCUSSIONS
without snow.
The mean annual temperature is 12.0°C and the All four rootstocks come from the
total active temperature is 3988°C, out of Mediterranean, are used in countries like Italy,
which 3170°C during the growing season; the France, Spain, Turkey.
115
Ishtara - is a French rootstock that has medium- susceptible to Pseudomonas syringae and
to-small vigour, it ensures precocity and good resistant to root-knot nematodes (Meloidogyne
fruit quality; it does not ensure proper tree spp).
anchoring and has no affinity with all apricot Resistant to calcareous soils (8-9% active
cultivars. lime). Suitable for replanting. All plum
Weiwa - Prunus domestica rootstock obtained cultivars shows perfect grafting compatibility
from the cultivar ‘Wangenheims’. This and high yield efficiency. With plum cultivars,
roostock is very resistant to “apricot and plum this rootstock reduces vigour by 15-20%
decline” and is tolerant to phytoplasma disease. compared to ‘Myrobalan B’. Many apricot
Trees grafted onto Weiwa rootstock have 30% cultivars rootstocks have been breeded and are
less vigour compared to Myrobolan 29 C spread out without knocking their behavoir in
(http://www.vitroplant.it/weiwa/?lang=en). fruit tree nursery (Field I and Filed II)
It has great affinity with all apricots and plums. regarding the grafting compatibility, scion
Weiwa has a very positive impact on both growing and the productivity.
cropping efficiency and fruit size. The The percentage of grafting was 62% at Adesoto
rootstock is not suckering. up to 98% at Ishtara (Table 1).
Adesoto - Selection of Pollizo de Murcia Regarding the trunk diameter, the lowest value
obtained in Spain. Interesting for the positive was recorded at Ishtara (11.85 mm) and the
grafting affinity and adaptability to very highest value (18.57) was found at Myrobolan
calcareous and dry soils. Tolerates to 29 C cultivar (Table 2).
Armillaria root-sucking. In Field II the height of the trees recorded was
Myrobolan 29 C - Clone selected from a 146 cm at Ishtara and 180 cm at Myrobolan 29
progeny of Prunus cerasifera in California. C.
Rootstock for plums and most apricot cultivars. The number of anticipated shoots varies from 3
Suitable for all soil types, adapts well to dry to the Weiwa cultivar up to 7 for the Ishtara
soils and heavy soils with low permeability. cultivar. The number of fruit buds ranges from
Moderately resistant to Agrobacterium 4 to Adesoto and 9 up to Ishtara.
tumefacens, Verticillium and Leptonecrosis,
Table 1. Number of trapped eyes and percentage of grafting, RSFG Constanta

APRICOT
Nr. eyes caught in
No. The number of grafted Percentage of
Rootstocks Variety the first inventory
seedlings (August 2015) grafting
(April 2016)
1. ADESOTO 50 31 62
2. ISHTARA 50 49 98
ʻAmiralʼ
3. MYROBOLAN 29 C 50 48 96
4. WEIWA 50 42 84
*The used grafting height was 5 cm above the soil.
Table 2. Behavior of some apricot rootstocks in field II of RSFG nursery Constanta, 2016
Number of anticipations Number of fruit
No. Rootstocks Cultivar Ø (mm) H (cm)
(pieces) buds (pieces)
1 ADESOTO 16.02 163 4 4
2 ISHTARA Amiral 11.85 146 7 9
3 MYROBOLAN 29 C 18.57 180 5 5
4 WEIWA 17.56 171 3 6
Average 16.00 165 4.75 6.00
116
Figure 4. Ishtara rootstock at the planting time
Figure 1. Diameter of trunck in second year CONCLUSIONS

at Myrobolan 29 C
Grafting percentage was 98% for Ishtara
variety.
The Ishtara cultivar shown the lowest value for
the thickness of the trunk.
The number of anticipated shoots is higher for
the Ishtara cultivars.
The number of fruit buds is higher for the
Ishtara cultivars.
The lowest height compared to the other
rootstocks was registered with the Ishtara
variety.
REFERENCES
Bălan et al., 2008. Caisul. Editura Ceres, București, 686.
Cociu et al., 2003. Caisul. Editura Ceres, Bucuresti, 438.
http://www.vitroplant.it/weiwa/?lang=en
Figure 2. Tree height in second year at Adesoto rootstock
Figure 3. Adesoto rootstock at planting in nursery
117
118
PRODUCTIVITY OF GOOSEBERRY VARIETIES

IN THE REPUBLIC OF MOLDOVA CONDITIONS
Parascovia SAVA
Scientifico-Practical Institute of Horticulture and Food Technology,

14 Costiujeni Street, Chişinău, MD 2019, Republic of Moldova

Abstract
In the paper are reflected the results of the scientific researches carried out in Republic of Moldova regarding the
determination of the phonological phases of the cultivation of gooseberry, the productivity and the exploitation period
of the intensive plantation established in 2004 with the ‘Colobok’, ‘Captivator’, ‘Grushenka’, ‘Sadko’, ‘Smena’,
‘Severnîi capitan’ varieties during the years 2015-2017 on irrigated land. Among the studied gooseberry varieties with
few thorns, resistant to mildew, low temperatures and drought, showed great fruits are: ‘Smena’ (3.5 g), Colobok (3.4
g) and with small fruits: ‘Severnîi capitan’ (1.7 g) and ‘Grushenka’ (1.9 g). The average weight of the gooseberry fruits
through the studied varieties ranged between 1.4 g and 3.3 g. The average harvest of the varieties studied grew between
6.2-18.1 t/ha.
Key words: gooseberry, fruits weight, phenophases, variety, yield.
INTRODUCTION Gooseberry productivity vary strongly

depending on climatic conditions specific to the
Variety occupies a central place in increasing year, characteristic features of varieties and
productivity, being an independent and abso- may decrease from 12 t/ha to over 1 t/ha
lutely determined factor. On the basis of a more (Olhina, Riabuschina,1987).
productive variety, without additional costs, it Gooseberry harvest depends on the genetic
is possible to increase the crop yield 1.5 times potential of the variety, the climatic conditions
and more. An intensive variety of gooseberry established during the growing season of each
must combine a whole range of qualities such year and their pollination by the bees. The
as: the adaptation reaction to the conditions of productivity of gooseberry plants is influenced
the growing medium, in case it does not adapt directly by the variety (Kawecki, 1989).
well, the accumulated energy is spent on Form the productivity point of view, the stu-
adaptation and not on the formation of the crop, died gooseberry varieties were divided in four
the poor resistance to diseases and pests can categories: low productivity (‘Grushenka’ - 1.2
destroy more than 1/3 of the crop, early entry t/ha), medium productivity (ʻZenitʼ - 3.5 t/ha,
bearing, self-fertility, which guarantees harvest ʻSomeşʼ - 3.3 t/ha, ʻOrlionokʼ, ʻFinikʼ - 2.8
in unfavorable conditions for flowering, fruit t/ha), high productivity 5.5 - 4.2 t/ha - ʻScedrâiʼ,
quality, cold resistance, drought and the final ʻRuskiiʼ, ‘Smena’, and very high productivity:
result - the crop (Sergheeva, 1985). ʻSevernîi capitanʼ - 14.6 t/ha (Sava, 2010).
In order to optimize the fuller realization of the The varieties with vigorous growth as ‘Cap-
productive potential, it is necessary to know the tivator’, ‘Severnîi capitan’, ‘Colobok’, under
tolerance potential of the variety for cultivation irrigation conditions, in intensive plantations
in different ecological areas, the conditions of (1.5 x 1.0 m) can give a high average yields
which correspond better to the requirements of ranging from 13.8 to 19.5 t/ha (Sava, 2012).
the plantation. Although the level of resistance,
which is specific to each species, variety, or MATERIALS AND METHODS
even plant, is a hereditary, genetically control-
led character, under optimal conditions it is The investigations were carried out during the
latent, unrealized, and only occurs under years 2015-2017 according to the methods
extreme stress conditions (Ştefârţă, Alunici et established studying shrubs and influenced the
al., 2001).
119
cultivation conditions in determining the days and the flowering started on 03.04.2016.
duration of the phenological phases of develop- At the end of March there were frosts higher
ment of the gooseberry plants, the fruit mass than -5 degrees C, which affected the first
and the productivity of the intensive plantation, young leaf of goosebrry, a phenomenon that
established in the year 2004 with a planting has not happened in the past. In 2017, the
distance of 1.5 x 1.0 m, on irrigated land, with vegetation period began earlier, similar to
varieties which have a few thorns and are 2016, and the budding took place on 08.03., the
resistant to mildew: ‘Smena’, ‘Sadko’, period until the flowering lasted 32 days and
‘Colobok’, ‘Captivator’, ‘Severnîi capitan’, the flowering began on 09.04.2017. At the end
‘Grushenka’. of April (19-22) frosts took place and wet snow
fell with a layer of 50 cm, which affected the
RESULTS AND DISCUSSIONS harvest, the plants being with formed small
fruits.
Climate change and its influence on the
cultivation conditions of shrub species becomes
more evident. The climatic conditions during
the 2017 vegetation period were manifested by
lower temperatures than in the previous years,
starting in the third decade of April, which has
led to a delay in going through the phenological
phases of the development of gooseberry
plants, and the rains from the end of May to the
beginning of June and the low temperatures at
night delayed the growth of plants.
The observations made during the evolution of
the phenological phases of gooseberry during
the vegetation period, 2015-2017, were Figure 1. Climatic conditions during the vegetation
introduced in Table 1. period, 19-22.04.2017
Table 1. The duration of the phenological phases of

gooseberries plants
Name of Years Ave
phenophases, rage
duration between
them (days) 2015 2016 2017
Budding date 16.03 06.03 08.03
Duration between 34 34 32 33
phases (days)
Blooming 19.04 03.04 09.04

Figure 2. Growing of gooseberry fruits
Duration between 59 72 73 68
phases (days)
The productivity of gooseberry varieties
Ripening 17.06 19.06 21.06 studied per hectare depends on the climatic
conditions of the year, the characteristics of
each variety, the capacities to adapt to the new
As can be seen from Table 1 data, the duration cultivation conditions, crop maintenance, the
between budding and blooming phases in 2015 age of plantation etc.
reached 34 days. In 2016, the vegetation period The study of the gooseberry varieties
began earlier, similar to the year 2002, when introduced during 2007-2017 allowed their
positive temperatures were established at the appreciation by mass of the fruit and the
end of February and the budding took place on production obtained, and the results are
06.03, the period until the flowering lasted 34 included in Table 2.
120
Table 2. Production and average mass of gooseberries
fruits depending on variety and year conditions
Average
2015 2016 2017
Variety fruit fruit fruit fruit
yield, yield yield, yield,
mass, mass, mass, mass,
t/ha t/ha t/ha t/ha
g g g g
Colobok 2.5 8.9 1.9 6.0 3.4 12.7 2.6 9.2
‘Captivator’ 2.2 19.2 2,9 18.0 2.1 17.1 2.4 18.1
‘Sadko’ 3.0 6.7 2,7 6.7 3.1 6.7 2.9 6.7
ʻSevernîi 1.4 15.9 1,7 22.0 1.7 15.1 1.6 17.7 Fig. 4. Gooseberry varieties with high productivity -
capitanʼ ‘Captivator’
‘Smena’ 3.0 4.7 3,4 6.7 3.5 7.3 3.3 6.2
1.4 11.9
CONCLUSIONS
‘Grushenka’ 1.1 14.0 - - 1.7 9.8
1.1- 4.7- 1.7- 6.0- 1.7- 6.7- 1.4- 6.2- On the basis of the research carried out on the
Average 3.0 19.2 3.4 22.0 3.5 17.1 3.3 18.1 gooseberry varieties introduced to the
pedoclimatic conditions of the Republic of
Moldova, it was established that:
As shown in Table 2, the average weight of The phenological phases of the development of
the fruits of gooseberry varieties ranged from the gooseberry plants are influenced by the
1.4 to 3.3 g values. The largest fruits were climatic conditions, and on average between
obtained in 2017, the maximum mass ranged the stages of budding and blossoming there is a
from 1.7 g to 3.5 g. period of 33 days, and between the flowering
The varieties with large fruits are: ‘Smena’ and the maturing of the fruit the interval is 68
(3.5 g), ‘Colobok’ (3.4 g), ‘Sadko’ (3.1 g). The days. The average weight of gooseberry fruits
varieties with small fruits are: ‘Severnîi capitan’ varied between 1.4 g in the ‘Grushenka’ variety
(1.7 g) and ‘Grushenka’ (1.9 g). The average of and 3.3 g in the ‘Smena’ variety.
fruit weight ranged from 1.4 g (variety The average fruit harvest obtained in the goose-
‘Grushenka’) to 3.3 g (variety ‘Smena’). berry varieties ranged between 6.2 (‘Smena’
The average fruit harvest obtained for the variety) and 18.1 t/ha (‘Captivator’ variety).
gooseberry varieties studied ranged between 6.2
(variety ‘Smena’) and 18.1 t/ha (variety REFERENCES
‘Captivator’). The maximum yield of 22.0 t/ha
of gooseberry fruits was obtained in the 2016 Kawecki Z., 1989. Wzrost I plonowanie krzewow dwoch
year for the variety ‘Severnîi capitan’, and in odmia agrestu wwarun Kach Olztynai Tolkmicka.
2015 year - 19.2 t/ha for the ‘Captivator’ variety. Acta Acad. Agricult. Techn. Olst. Agric. Nr. 13,
189-198.
Olihina E.I., Riabushkina E.V., 1987. Sortoizucenie
iagodnâh culitur. Sb. Naucinah trudov., vap. 49,
Vsesoiuznai NII sadovodstva im. I.V. Miciurina,
Miciurinsc, 11-16.
Sava P., 2010. Specific caracteristics of gooseberry va-
rieties in new cultivation conditions. Scientific
papers Journal, Horticulture Series, LIII, „Ion
Ionescu de la Brad” Publishing House, Iaşi, 235-240.
Sava P., 2012. Bazele știintifice ale culturii agrișului în
Republica Moldova. Monografie. Tipografia UASM,
Chișinau, 192.
Ştefârţă A., Alunici N., Buceaţcaea S., Brânză L., Vrabie
V., 2001. Metodele noi de diagnosticare şi
ameliorare a rezistenţei plantelor la un deficit
moderat de umezeală. Fiziologia şi biochimia
plantelor la început de mileniu: Realizărişi
Fig. 3. Gooseberry varieties with high profuctivity -
perspective. Mater. Conr. II, Ch., 342-348.
ʻSevernîi capitanʼ
121
122
STUDIES ON IN VITRO FERTILIZATION BIOLOGY

OF MESPILUS GERMANICA L. CV. ʻİSTANBULʼ
Sultan Filiz GÜÇLÜ1, Hasan BEKTAŞ2, Fatma KOYUNCU2

1
Suleyman Demirel University, Atabey Vocational School, Isparta, Turkey
2
Suleyman Demirel University, Faculty of Agriculture, Horticultural Science,
32260, Isparta, Turkey
Abstract
This study was aim to investigate the fertilization biology of ʻİstanbulʼ medlar variety. For this purpose, firstly pollen
production capacity and morphological homogeneity were assessed (hemocytometer slide). The pollen viability was
tested with TTC and IKI staining methods. Pollen germination experiments were carried out in different medium with
different sucrose concentration (agar in plate method) during 2 h, 6 h, 12 h and 24 h period. The number of anthers in a
flower 36.5, pollen number in an anther 160.55 and pollen number in a flower 12753.8 were found. Morphological
homogeneity was high with 98%. Pollen viability rates varied between 94% (TTC) and 96% (IKI). Contents of medium
V: 20% sucrose 1% agar-agar+5 ppm boric acid and 24 hours incubation duration were stated as optimum pollen
germination rate and tube growth.
Key words: Medlar, pollen germination, morphological homogeneity, pollen tube growth, TTC.
INTRODUCTION Moore, 1996; Tosun and Koyuncu, 2007). The

first condition of formation of seed and fruit is
Medlar (Mespilus germanica L.) has been developing healthy male and female organs of
cultivated for over thousands of years in the flower and cells, except for an apparent
temperate zones of Anatolia. The medlar, partenocarpy of some cultivars. Pollen
called as ‘Muşmula’, ‘Beşbıyık’ or ‘Döngel’ performance, which includes pollen
(in Turkish), is botanically classified as a germination, pollen tube growth rate and pollen
pome and produces edible fruits (Atay, 2013). competition, is an important component of
Mespilus germanica L. belongs to Rosaceae fertilization success in seed-producing plants.
family and it grows mainly in frost-free areas, Pollen performance is clearly affected by the
and on rocks and poor soils (Haciseferoǧullari genotype of the pollen (Acar and Kakani, 2010;
et al., 2005). The flowers are hermaphrodite, Hedly et al., 2004). There have been some
five-piece, white-pink color, and each bud has studies about ‘İstanbul’ medlar, but apparently
one flower. Flowers generally open in May- almost no work has been done on fertilization.
June. In Turkey, they are abundant The aim of this study is to examine the pollen
particularly in north and west-Anatolia and performance of the ʻİstanbulʼ medlar variety
Marmara regions. Medlar is one of the latest and determine the optimum pollen germination
maturating fruits and the ripening occurs in medium protocol.
late October before frosts in Turkey. The
fruits are used as a nutrition component by the MATERIALS AND METHODS
local population and are prepared by the local
people as marmalade or pickle (Ercisli et al., In the study, pollens taken from 'İstanbul'
2012). medlar trees in the orchard of Eğirdir Fruit
Fertilization success in plants is the result of Research Institute were used. Pollens were
processes that take place during the progamic obtained from flowers at balloon stage. The
phase (Thompson, 2004; Güçlü and Koyuncu, flowers were transferred to the laboratory
2017). Pollen germination and pollen tube immediately. Anthers were removed and placed
growth are important components of into the dark-colored bottle to promote
fertilization success in fruit trees (Janick and dehiscence at room temperature. For the pollen
123
performance, pollen production capacity, germination and pollen tube growth
morphological homogeneity, pollen viability experiments. For each assay, 2 mL of medium
rates were investigated. In addition, optimum was placed into Petri dishes. Counts were made
germination medium and incubation period from 4 different microscope fields (100-150
were determined. The pollen production pollen grains per field for each Petri dishes)
capacity and morphological homogeneity (Hedly et al., 2004; Koyuncu, 2006). Statistical
percentages of pollens were assessed with the analysis was conducted using Duncan’s
hemocytometer (Marienfeld, Germany) slide multiple range test within the general linear
(Eti, 1990). Imperfectly shaped pollen grains model procedure of SPSS 16.0.
were considered as aborted pollen. The final
percentage of morphological homogeneity RESULTS AND DISCUSSIONS
(MH) was defined as:
Pollen production capacity and pollen
ܰ‫ ݏ‬െ ܰܽ viability
‫ ܪܯ‬ൌ ‫ͲͲͳݔ‬
‫ܣ‬ Pollen production amount and viability results
were shown in Table 1. Pollen morphological
Ns: Number of shaped pollen per area; homogeneity (98%) and pollen viability (96%)
Na: Number of aborted pollen per area; were found very high IKI viability test (96%)
A: Total number of pollen area. was higher than TTC (94%) test. Similarly,
researcher reported that pollen viability in the
In the stain tests, pollen viability was estimated IKI test was generally higher (84.83-100%) and
by using TTC (2, 3, 5-triphenyl tetrazolium stable than TTC test (11.78-91.66%)
chloride) and FDA (fluorescein di acetat) (Çavusoğlu and Sülüsoğlu, 2013). Koyuncu
stains. Pollens were scattered onto TTC and (2006) studied strawberry pollens using TTC
FDA solutions, and stained pollens were and reported that pollen viability ratios reached
counted after 2 hours and 15 minutes, 82% (Allstar and Elvira) and 86.5%
respectively. To determine the pollen viability, (Chandler). Koyuncu and Tosun (2009) used
pollens of each cultivar (of four different areas) TTC, FDA and IKI stain tests for the same
were observed onto two slides under a light sweet cherry cultivars. They reported that the
microscope (×100 magnification). The stained pollen viability differed according to stain
pollen was considered as viable in these tests. methods and cultivars.
Four concentrations of sucrose were tested in
the germination medium for the in vitro Table 1. Pollen production capacity, morphological
germination test. homogeneity and pollen viability tests
I. 5% sucrose + 1% Agar-Agar + 5 ppm
H3BO3 (Boric Acid)
II. 10% sucrose + 1% Agar-Agar + 5 ppm
H3BO3
III. 15% sucrose + 1% Agar-Agar + 5 ppm
H3BO3 N: Numbers of anthers in a flower; M: Mean pollen
IV. 20% sucrose + 1% Agar-Agar + 5 ppm number in an anther; PN: Pollen number in a flower;
MH: Morphological homogeneity.
H3BO3
The ‘agar in plate’ method was used to
establish pollen germination and pollen tube Pollen germination tests
growth (Koyuncu and Tosun, 2009). Pollen For the optimum germination medium different
tube long at least as its diameter was sucrose concentrations were tried.
considered to be ‘germinated’. The percentage A linear equation best described the
of pollen germination was determined after 2 h, relationship between the germination
12 h and 24 h incubation period at 21°C. An percentage and different concentration of
ocular micrometer was used to measure pollen sucrose (r 0.966). In Medium I, sucrose
tube length, under a light microscope, at a concentration was the lowest, pollen
magnification. Four Petri dishes were used for germination has not started in 2 hours whereas
the others even low gave germination at 2
124
hours with increasing sucrose concentration. As When different incubation periods are
seen Figure 1, pollen germination rates compared in the same germination medium, the
increased with increasing sucrose difference between the values is statistically
concentration. The highest ratio of pollen significant (p<0.05). Pollen germination raised
germination was obtained from Medium IV as much as 3 fold between 6 h and 12 hours
(20% sucrose + 1% Agar-Agar + 5 ppm reached own maximum percentage at 24 hours
H3BO3). In terms of mean pollen germination (10.1; 22.3; 54.9; 86.8, respectively) for all
rate, it followed by Medium III (16.82%) and mediums. Results of incubation duration
Medium II (7.25) (Table 2). This suggests that experiments were similar to the findings of
the increased concentration of sucrose Yıldız and Yılmaz (2002), who reported that
increases the germination rate. Çavuşoğlu and the germination pollen of strawberry cultivar
Sülüsoğlu (2013) reported that pollen ʻTuftsʼ began within 1 h at 24°C. Tosun and
germination varied between 16.4%-66.67% for Koyuncu (2007) also reported that the
all germination media. Another study which germination rates increased with incubation
was conducted in Arabidopsis emphasized period in cherries.
sugar-dependent multilayer regulation of
Arabidopsis pollen germination is supported, Table 2. In vitro pollen germination percentages of
which makes this approach a valuable medlar in different sucrose concentrations and incubation
periods
experimental system for future studies
addressing sugar sensing and signaling
(Hirsche et al., 2017). Güçlü and Koyuncu
(2017) found the suitable germination medium
0.5% agar + 15% sucrose + 5 ppm H3BO3
(Boric acid) for apple’s pollen.
*Values within a column followed by different letters are
significantly different (p<0.05).
When the data were evaluated, in a medium

with different sucrose concentrations, a linear
increase was observed between the rate of
germination and the duration of incubation
(Figure 2).
Figure 2. Linear regression curves between germination

rate and incubation period
The in vitro elongation of pollen tubes was

Figure 1. In vitro pollen germination percentages of affected by different sucrose concentration
medlar at 2 h, 6 h,12 h and 24 hours (p<0.05) at different (Table 3). The effect of different concentrations
sucrose concentration (a); Regression curve between
germination rate and sucrose concentration (b)
of sucrose on the pollen tube elongation was
statistically significant (p<0.05). As well as the
pollen germination rate, pollen tube growth
125
increased during the incubation period and (Mespilus germanica L.), Journal of Applied Botany
measured at 24 hours later. As the sucrose and Food Quality 85 (1): 86-90.
Eti S., 1990. Cicek tozu miktarini belirlemede kullanilan
concentration increased, the length of the pratik bir yontem. Journal of Agricultural Faculty,
pollen tube increased, reaching the highest Cukurova University, 5: 49-58.
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126
RELATIONSHIP BETWEEN VEGETATIVE GROWTH AND NUT

CHARACTERISTICS IN ALTERNATE BEARING PISTACHIO (PISTACIA
VERA) CULTIVARS EXPOSED TO DROUGHT
Hakan CETINKAYA1*, Muhittin KULAK2, 3, Ahsen Işık OZGUVEN4

1
Kilis 7 Aralık University, Faculty of Agriculture, Department of Horticulture, Kilis, Turkey
2
Kilis 7 Aralık University, Faculty of Arts and Sciences, Department of Biology, Kilis, Turkey
3
Department of Nursing, Yusuf Serefoglu School of Health, Kilis 7 Aralık University, Kilis, Turkey
4
Department of Plant Production and Technologies, Faculty of Agricultural Sciences and
Technologies, Cyprus International University, Nicosia, Turkish Republic of Northern Cyprus
Corresponding author email: [email protected].
Abstract
Among abiotic stress factors, drought stress is of the most influential factors limiting plant growth and impairing plant
metabolism. In addition to the abiotic stress factors, alternate bearing is a main physiological problem for horticultural
plants including olive, pistachio, hazelnut and apple. In this context, a 3-year field study was designed for three
pistachio cultivars, namely ‘Uzun’, ‘Siirt’ and ‘Ohaidi’. The cultivars were exposed to irrigated and non-irrigated
conditions. After experimental period, yield, shoot length and nut characteristics were measured. According to years,
shoot length, 100 nut weight, nut internal weight, nut length, nut width and nut thickness decreased but blank nut ratio
increased in productive year as a yield. Regarding with varieties, there was no significant difference yield, blank nut,
and split nuts among cultivars but shoot length, nut width and thickness were higher in ‘Ohaidi’ cultivar. Lowest 100
nut weight and nut internal weight were determined in ‘Uzun’ cultivar and highest nut length was observed in ‘Siirt’
cultivar. Irrigation only affected the shoot length from given properties. As a result, yield of the tree affected vegetative
development and nut characteristics. Along with this study, finally vegetative growth and fruit characteristics were
correlated.
Key words: alternate bearing, pistachio cultivars, Pistacia vera, vegetative growth, nut characteristics.
INTRODUCTION trees, inadequate pollination, traditional

cultural practices, fertilization, harvesting,
Turkey has an important place in the world prolonged period of water stress etc. (Elloumi
with its unique ecological conditions and is the et al., 2013; Acar and Eti, 2007; Khoyerdi et
gene center of pistachio and pistachio al., 2016; Marcinska et al., 2013; Dag et al.,
production, spreading in the south-eastern parts 2009; Kallsen, 2017). Disturbance in adequate
of Turkey. The irregular fruiting behaviour and balanced nutrition of the plant lead to low
(alternate bearing) and its effects in yield productivity and quality. The alternate bearing
reduction in pistachio are of the major is the result of the disruption deviating from
problems. Alternate bearing is related to many optimal nutrition caused by domination of fruit
factors mainly classified as intrinsic factors in relation to nutrient use competition between
such as endogenous hormones, nutrition, fruit, buds and leaves (Crane and Nelson,
carbohydrate accumulation allocation between 1971). Pistachio is mainly grown under rain fed
reproductive and vegetative organs of the plant, conditions in the Mediterranean basin and
rootstocks, cultivars, genetic factors (Nzima et considered as drought and saline-resistant crop
al., 1997; Spann et al., 2008; Kumar et al., (Behboudian et al., 1986; Rieger, 1995).
2016; Goldschmidt and Golomb, 1982; Irrigation is of the essential factor for plant
Rosecrance et al., 1998; Durand et al., 2013; yield. In pistachio, irrigation improves nut
Kafkas et al., 2006) and extrinsic such as quality and reduces the alternate bearing
environmental, climatic and soil properties, pattern (Kanber et al., 1993).
climatic changes in rainfall and temperature, Plenty of flower buds are formed every year in
compensate chilling and total heat demand of pistachio that makes it different from other
127
species showing alternate bearing. However, Statistical analysis:
the flower buds fall off during the summer SPSS statistical program was used to determine
months. statistical significance levels by employing the
The alternate bearing in pistachio is the result independent one-way ANOVA followed by
of flower buds falling off, not concerned with Duncan multiple range test and the differences
flower bud formation (Crane, 1971). between individual averages were considered to
Of the abiotic stress factors, drought is be statistically important at p< 0.05. The results
considered as the most hazardous factors on were expressed as mean.
sustainable agriculture worldwide (Din et al.,
2011; Anjum et al., 2012; Lefèvre et al., 2012). RESULTS AND DISCUSSIONS
Drought is assumed to be the consequences of
soil and/or atmospheric water deficit, causing The yield, vegetative growth and nut
substantial eco-physiological constraints to characteristics values of ‘Uzun’, ‘Siirt’ and
plant survival and to crop productivity and ‘Ohaidi’ pistachio varieties were measured for
quality (Boyer, 1982). three years on irrigated and non-irrigated
Drought associated studies are great concern of conditions and the inter-parameter relationships
the plant scientists for developing, improving were determined by statistically evaluating the
or exploring the drought-tolerant plant species changes between parameters according to
(Hamrouni et al., 2001; Al-Barrak, 2006; years, irrigation and varieties. The obtained
Bettaieb et al., 2009; Bybordi, 2010). values and changes are shown in Table 1.
In the current study, it was aimed to reveal the Changes in yield, growth and nut
relationship between yield and vegetative characteristics’ parameters according to
growth and nut characteristics in pistachio irrigation, cultivars and years are represented in
cultivars bearing alternatively in response to Table 2. Accordingly, irrigation increased shoot
the irrigated and non-irrigated conditions. length. Of the varieties, ‘Ohaidi’ with more
shoot, ‘Siirt’ and ‘Ohaidi’ with more 100-nut
MATERIALS AND METHODS weight and internal nut weight, ‘Siirt’ with
broader nut and ‘Ohaidi’ with thicker nuts were
Experimental Material of the results of the present study.
The research was carried on 25 years old When differences between the years were
cultivars ‘Siirt’, ‘Uzun’ and ‘Ohaidi’ grafted investigated, the yields of the following years
onto a Pistacia vera rootstocks which were were about the same, while the first year
cultivated in 7 x 2 m intervals at Pistachio received more yield than the other years.
Research Institute. During the year with fewer yields, higher shoot
The experiments were designed under irrigated length and lower blank nut, 100-nut weight,
and non-irrigated conditions according to the internal nut weight, split nut, nut length, width
randomized plot design with four replicates. and thickness were obtained.
Changes and correlations between the yield,
Yield, vegetative growth and nut shoot growth and nut characteristics under
characteristics irrigated and non-irrigated conditions for
Yield (kg/tree) was obtained by dividing the ‘Uzun’ cultivar are shown Table 3 and Figure
total harvested product per tree for each 1. In terms of correlation coefficient (r) values
variety. there were negative correlation between yield
Shoot length (SL) was found by measuring the
and blank nut under non-irrigated (-0.811)
shoots at the end of vegetation. Counts and
condition, yield and 100-nut weight under
measurements of the fruits in the clusters were
made for the fruit characteristics. Nut length irrigated (-0.844), yield and shelling percen-
(NL), nut width (NW), nut thickness (NT), nut tages under irrigated (-0.888) condition, yield
weight (100-NW), internal nut weight (INW), and nut width under non-irrigated (-0.807)
shelling percentage (SP) and blank nut (BN) condition, yield and nut thickness under
are also evaluated for the fruit characteristics. irrigated (-0.974) and non-irrigated (-0.979)
128
Table 1: Growth and nut characteristics’ parameters under irrigated and non-irrigated conditions for pistachio cultivars
‘UZUN’ ‘SIIRT’ ‘OHAIDI’

1.Year 2.Year 3.Year 1.Year 2.Year 3.Year 1.Year 2.Year 3.Year
IR 4.6 2.9 3.3 5.6 3.7 3.8 4.5 3.1 4.8
Yield
N-IR 6.4 3.2 5.1 7.7 3.7 3.6 6.4 2.8 4.3
(kg/tree)
AVE 5.49 3.05 4.20 6.65 3.68 3.71 5.44 2.93 4.58
IR 12.8 13.0 21.5 16.6 18.6 22.5 25.0 25.9 32.0

Shoot lenght
N-IR 13.0 14.3 19.8 13.8 13.1 21.2 21.5 24.1 27.9
(cm)
AVE 12.90 13.65 20.65 15.20 15.85 21.85 23.25 25.00 29.95
IR 17.7 15.7 9.2 22.9 9.7 8.0 18.1 10.9 7.1

Blank Nut
N-IR 14.0 22.2 11.7 25.4 17.8 11.2 40.1 14.6 9.7
(%)
AVE 15.85 18.95 10.45 24.15 13.75 9.60 29.10 12.75 8.40
IR 168.5 181.2 187.7 187.0 249.7 252.3 198.4 244.6 249.9

Nut weight
N-IR 165.1 183.4 185.5 179.2 245.3 246.0 220.6 243.9 246.9
(g/100 nut)
AVE 166.8 182.3 186.6 183.1 247.5 249.2 209.5 244.3 248.4
IR 59.0 56.0 61.6 69.7 92.3 94.4 81.0 91.1 95.2

Internal nut weight
N-IR 61.4 56.9 59.6 74.4 88.8 89.7 90.6 93.5 93.2
(g/100 nut)
AVE 60.20 56.45 60.60 72.05 90.55 92.05 85.80 92.30 94.20
IR 30.0 51.5 59.0 36.0 68.5 80.0 26.0 76.5 77.0

Shelling
N-IR 57.5 56.0 61.5 32.5 65.0 75.0 30.0 43.5 74.0
(%)
AVE 43.75 53.75 60.25 34.25 66.75 77.50 28.00 60.00 75.50
IR 25.0 25.5 26.1 24.5 27.0 27.4 23.0 23.8 23.0

Nut lenght
N-IR 24.5 25.0 26.1 26.0 26.5 27.4 23.0 23.8 22.5
(mm)
AVE 24.75 25.25 26.10 25.25 26.75 27.40 23.00 23.80 22.75
IR 13.5 13.5 14.0 14.2 15.1 15.0 14.5 15.0 15.5

Nut width
N-IR 13.0 13.5 13.5 14.2 15.1 14.5 15.0 14.5 15.0
(mm)
AVE 13.25 13.50 13.75 14.20 15.10 14.75 14.75 14.75 15.25
IR 12.1 13.0 13.0 13.0 14.3 14.2 15.3 15.0 15.0

Nut thickness
N-IR 12.1 13.0 12.5 13.5 14.3 14.2 15.3 14.5 14.5
(mm)
AVE 12.10 13.00 12.75 13.25 14.30 14.20 15.30 14.75 14.75
Table 2: Changes in yield, growth and nut characteristics’ parameters according to irrigation, cultivars and years
Shoot Internal nut Nut Nut Nut

Yield Blank Nut weight Shelling
length weight length width thickness
kg/tree Nut (%) (g/100 nut) (%)
(cm) (g/100 nut) (mm) (mm) (mm)
IR 4 20.8 a 13.2 213.3 78.2 56.1 24.8 14.4 13.6

IRRIGATION N-IR 4.8 18.7 b 18.6 212.9 78.0 55.0 24.6 14.2 13.6
‘UZUN’ 4.2 15.6 b 15.1 178.6 b 59.2 c 52.6 25.2 b 13.5 b 12.4 c
‘SIIRT’ 4.7 17.6 b 15.8 226.6 a 84.9 b 59.5 26.2 a 14.4 a 13.8 b
CULTIVARS ‘OHAIDI’ 4.3 26.0 a 16.8 234.0 a 90.8 a 54.5 22.8 c 14.9 a 14.7 a
1.YEAR 5.9 a 17.1 b 23.0 a 186.5 b 72.7 b 35.3 c 24.2 b 14.0 b 13.3 b
2.YEAR 3.2 b 18.1 b 15.1 b 224.7 a 79.9 a 60.2 b 25.0 a 14.3 ab 13.8 a
YEARS 3.YEAR 4.2 b 24.1 a 9.6 b 228.0 a 82.3 a 71.1 a 24.9 a 14.6 a 13.8 a
129
condition, positive correlation between yield nut weight and shelling (0.981), nut length
and internal nut weight under irrigated (0.999) (0.894), width (0.951) and thickness (0.887),
condition. There was no particular correlation between shelling percentage and nut length
between yield and nut length. (0.964), width (0.993) and thickness (0.781),
between nut length and nut width (0.989) and
Table 3. Correlation coefficient (r) values of the growth thickness (0.989), between nut width and nut
and developmental parameters under irrigated and non-
irrigated conditions for ‘Uzun’ cultivar
thickness (0.700).
Y SH BN 100-NW INW SP FL FW shoot growth and nut characteristics under
IR -0.312 irrigated and non-irrigated conditions for
SL N-IR -0.073 “‘Siirt’” cultivar are shown Table 4 and Figure
AVE -0.136 2. Effects of yield on vegetative growth and nut
IR 0.500 -0.979 characteristics were more significant in ‘Siirt’
BN N-IR -0.811 -0.525 cultivar than the ‘Uzun’ and ‘Ohaidi’ cultivars.
AVE -0.315 -0.898
IR -0.844 0.773 -0.887

Table 4. Correlation coefficient (r) values of the growth
100- and developmental parameters under irrigated and non-
N-IR -0.748 0.716 0.218
NW irrigated conditions for ‘Siirt’ cultivar
AVE -0.775 0.731 -0.355
IR 0.565 0.834 -0.703 0.294 Y SH BN 100-NW INW SP FL FW

IR -0.729
INW N-IR 0.999 -0.066 -0.815 -0.744
SL N-IR -0.450
AVE 0.791 0.500 -0.830 -0.226
AVE -0.568
IR -0.888 0.714 -0.843 0.996 0.209
IR 0.989 -0.824
SP N-IR 0.366 0.901 -0.842 0.343 0.373
BN N-IR 0.895 -0.801
AVE -0.639 0.849 -0.828 0.981 -0.034
AVE 0.958 -0.779
IR -0.695 0.900 -0.970 0.972 0.510 0.948
IR -0.997 0.782 -0.998
NL N-IR -0.201 0.992 -0.411 0.800 -0.194 0.838
100-
N-IR -0.999 0.439 -0.890
NW
AVE -0.411 0.959 0.736 0.894 0.234 0.964
AVE -0.999 0.593 -0.967
IR -0.292 1.000 -0.974 0.760 0.845 0.700 0.891
IR -0.992 0.808 -1.000 0.999
NW N-IR -0.807 0.648 0.309 0.996 -0.803 0.254 0.741
INW N-IR -0.999 0.478 -0.909 0.999
AVE -0.541 0.907 -0.628 0.951 0.087 0.993 0.989
AVE -0.997 0.629 -0.977 0.999
IR -0.974 0.517 -0.682 0.943 -0.041 0.968 0.839 0.500
IR -0.955 0.899 -0.989 0.976 0.984
NT N-IR -0.999 0.117 0.784 0.777 -0.999 -0.325 0.244 0.832
SP N-IR -0.979 0.623 -0.967 0.976 0.985
AVE -0.979 0.333 0.117 0.887 -0.651 0.781 0.989 0.700
AVE -0.969 0.754 -0.999 0.976 0.985
IR -0.985 0.837 -0.999 0.996 0.999 0.992
Considering average value there were no NL N-IR -0.786 0,906 -0.979 0.779 0.805 0.896
correlation between yield and shoot length, AVE -0.953 0.790 -0.999 0.962 0.973 0.998
blank nut, shelling percentage, nut length and IR -0.999 0.690 -0.979 0.991 0.984 0.937 0.974
nut width but there were correlation 100-nut NW N-IR -0.742 -0.265 -0.365 0.750 0.721 0.589 0.169
weight (-0.775), internal nut weight (0.791) and AVE -0.926 0.215 -0.780 0.914 0.895 0.804 0.768
nut thickness (-0.979). IR -1.000 0.713 -0.985 0.995 0.989 0.948 0.981 0.999
As regards to correlations between each other NT N-IR -0.991 0.325 -0.827 0.992 0.986 0.942 0.695 0.826
except yield, correlations were found between AVE -0.997 0.502 -0.933 0.994 0.988 0.947 0.927 0.952
shoot length and blank nut (-0.898), 100-nut

weight (0.731), shelling (0.849), nut length According to correlation coefficient (r) values
0.959) and nut width (0.907), between blank there were negative correlations between yield
nut and internal nut weight (-0.830), shelling and shoot length under irrigated (-0.729), yield
(-0.828) and nut length (0.736), between 100- and 100-nut weight under irrigated (-0.997) and
130
non-irrigated (-0.999) condition, yield and and blank nut weight under non-irrigated
internal nut weight under irrigated (-0.992) and (0.837) condition, yield and nut width under
non-irrigated (-0.999) condition, yield and non-irrigated (0.814) condition, yield and nut
shelling percentages under irrigated (-0.955) thickness under non-irrigated (0.910) condition.
and non-irrigated (-0.979) condition, yield and Considering average value there were correla-
nut length under irrigated (-0.985) and non- tions between yield and 100-nut weight (-0.696),
irrigated (-0.786) condition, yield and nut width nut length (-0.814) and thickness (0.763).
under irrigated (-0.999) and non-irrigated (-
0.742) condition, yield and nut thickness under Table 5. Correlation coefficient (r) values of the growth
irrigated (-1.000) and non-irrigated (-0.991) and developmental parameters under irrigated and non-
irrigated conditions for ‘Ohaidi’ cultivar
condition, positive correlation between yield
and blank nut weight under irrigated (0.989) Y SH BN 100-NW INW SP FL FW
and non-irrigated (-0.895) condition. There was IR 0.541
no particular correlation between yield and nut SL N-IR -0.490
length. AVE -0.075
Considering average value there were correla- IR 0.011 -0.835
tions between yield and blank nut (0.958), 100- BN N-IR 0.837 -0.887
nut weight (-0.999), internal nut weight (- AVE 0.618 -0.830
0.997), shelling percentage (-0.969), nut length IR -0.261 0.671 -0.968
(-0.953), width (-0.926), and thickness (-0.997). 100-

NW
N-IR -0.862 0.864 -0.999
As regards to correlations between each others AVE -0.696 0.768 -0.995
except yield, correlations were found between IR -0.073 0.799 -0.998 0.982
shoot length and blank nut (-0.779), shelling INW N-IR -0.945 0.748 -0.970 0.980
(0.754) and nut length (0.790), between blank AVE -0.605 0.839 -1.000 0.993
nut and 100-nut weight (-0.967), internal nut IR -0.342 0.606 -0.943 0.996 0.962
weight (-0.977), shelling (-0.999), nut length (- SP N-IR -0.389 0.994 -0.830 0.803 0.669
0.999), width (-0.780) and thickness (-0.993), AVE -0.516 0.893 -0.992 0.974 0.994
between 100-nut weight and internal nut weight IR -0.986 -0.394 -0.176 0.417 0.237 0.493
(0.999), shelling (0.976), nut length (0.962), NL N-IR -0.531 -0.479 0.018 0.028 0.225 -0.573
width (0.914) and thickness (0.994), between AVE -0.840 -0.479 -0.093 0.195 0.076 -0.032
internal nut weight and shelling (0.985), nut IR 0.165 0.919 -0.984 0.909 0.972 0.870 0.001
length (0.973), width (0.895) and thickness NW N-IR 0.814 0.108 0.364 -0.407 -0.579 0.218 -0.924
(0.988), between shelling percentage and nut AVE 0.179 0.968 -0.663 0.582 0.675 0.751 -0.684
length (0.998), width (0.804) and thickness IR 0.350 -0.599 0.940 -0.996 -0.960 -1.000 -0.500 -0.866
(0.947), between nut length and nut width NT N-IR 0.910 -0.807 0.989 -0.995 -0.996 -0.736 -0.132 0.500
(0.768) and thickness (0.927), between nut AVE 0.763 -0.702 0.980 -0.995 -0.976 -0.947 -0.289 -0.500
width and nut thickness (0.952).
As regards to correlations between each others
shoot growth and nut characteristics under
except yield, correlations were found between
irrigated and non-irrigated conditions for
shoot length and blank nut (-0.830), 100-nut
‘Ohaidi’ cultivar are shown Table 5 and Figure
weight (0.768), internal nut weight (0.839),
3. According to correlation coefficient (r)
shelling (0.893), nut width (0.968) and
values there were negative correlations between
thickness (-0.702), between blank nut and 100-
yield and 100-nut weight under non-irrigated (-
nut weight (-0.995), internal nut weight (-1.00),
0.862) condition, yield and internal nut weight
shelling (-0.992) and nut thickness (0.980),
under non-irrigated (-0.949) condition, yield
between 100-nut weight and internal nut weight
and nut length under irrigated (-0.986)
(0.993), shelling (0.974) and nut thickness
condition, positive correlation between yield
(-0.995), between internal nut weight and
131
shelling (0.994), nut width (0.675) and vegetative growth and little fruit on olive trees
thickness (-0.976), between shelling percentage (Lavee, 2007).
and nut width (0.751) and thickness (-0.947), Competition for energy resources between the
between nut length and nut width (-0.684). vegetative shoot meristem and fruit reduce
The reasons underlying alternate bearing are vegetative growth of the tree, so it designates
classified as (environmental, climatic and soil the reproductive status of the tree the following
properties) and intrinsic factors (nutritional year (Samach and Smith, 2013). For this reason
status, endogenous balance of hormone and in alternate bearing varieties cultural practices
interactions between organs). Extrinsic factors such as pruning, girdling, applications of plant
affect bearing alternatively to yield because of growth regulators and flower and fruit thinning
climatic changes in rainfall and temperature are to maintain a balance of vegetative and
(Elloumi et al., 2013). Intrinsic factors are reproductive shoots (Pellerin et al., 2011; Dag
related to endogenous hormones, nutrition and et al., 2009).
carbohydrate accumulation allocation between In pistachio nut, the nut splitting is a genetic
reproductive and vegetative organs of the plant characteristic and nut splitting ratio is affected
(Nzima et al., 1997; Spann et al., 2008). by rootstock, cultivar, plant nutrition, alternate
The results indicated that yield affected the bearing, climatic conditions, cultural manage-
vegetative growth and nut characteristics. In the ment and pollen source (Takeda, 1979; Crane
year of low yield, shoot length was increased et al., 1982).
accordingly blank nut, 100-nut and internal nut Percentage of split, non-split and blank nuts
weight, shelling percentage, nut length, width varied by shoot type, clusters harvested from
and thickness decreased. Irrigation increased long-shoots have higher total yield and split nut
shoot length. compared to short-shoot clusters because of the
Drought conditions decrease rate of cell locally higher carbohydrate supply from the
division and expansion, stem elongation, plant greater leaf area of the long-shoots (Kumar et
water and nutrient uptake and water use al., 2016). Because of photosynthesis and the
efficiency (Li et al., 2009), shoot and root dry transportation of in other photo-assimilate
weight, leaf relative water content, total (Marschner, 1995) to the developing nuts, 100-
chlorophyll, carotenoids and increased nut weight, split nuts, and internal nut weight
oxidative stress products, some osmoregulators increased.
and antioxidant agents (Khoyerdi et al., 2016; Nutrient status of trees and fertilization has
Marcinska et al., 2013; Shamshiri and Fattahi, effects on nut yield and quality in pistachio.
2014; Spann et al., 2009). Nitrogen, phosphorus, potassium and foliar
Because of competition between developing boron application influenced vegetative growth
fruit and vegetative growth, heavy fruit load measurements, the flowering and fruit set,
uses the plants of nutrients and carbohydrates reduced buds abscission, blank pistachios and
which are required for vegatative growth in turn improved nut quality characteristics
(Goldschmidt, 1999; Stevenson et al., 2000). A including, nut length, width and thickness
heavy fruit loaded branch will have little (Kumar et al., 2016).
Figure 1. Changes in growth and nut characteristics under irrigated and non-irrigated conditions for ‘Uzun’ cultivar
132
Figure 2. Changes in growth and nut characteristics under irrigated and non-irrigated conditions for ‘Siirt’ cultivar
Figure 3. Changes in growth and nut characteristics under irrigated and non-irrigated conditions for ‘Ohaidi’ cultivar
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134
EVALUATION OF POMOLOGICAL PROPERTIES OF SOME ORANGE

VARIETIES FROM WEST MEDITERRANEAN TURKEY
Gulay DEMIR1, Fatma KOYUNCU2

1
Bati Akdeniz Agricultural Research Institute, Antalya, Turkey
2
Suleyman Demirel University, Faculty of Agriculture, Isparta, Turkey
Abstract
Citrus trees can be productively grown in Mediterranean region of Turkey. In this study, six orange varieties
(Belladonna, Biondo, Biondo Riccio, Calabrese, Mediterranean Sweet, Parson Brown), grafted C. aurantium L. var.
ʻYerliʼ, were re-evaluated under Antalya ecological conditions. During the 2 consecutive trial years, fruit dimensions
(weight, height, width), rind thickness, seed number, total soluble solids (TSS), titratable acidity (TA), TSS/TA ratio,
fruit juice content, pH, vitamin C content were analyzed. There were statistically differences according to some
pomological properties of six varieties. Mediterranean Sweet has the biggest fruits (187.54 g). The highest TSS ratio
(11.0%) and vitamin C content (61.20 mg/100ml) was in Parson Brown orange. Biondo fruits have the lowest rind
thickness.
Key words: Citrus, C. sinensis, fruit quality.
INTRODUCTION potential to competing with other

Mediterranean countries in terms of quality
Mediterranean basin has suitable ecological (Tuzcu, 1998). It is important to evaluate well-
conditions to enable the edible production of adapted, productive and having good fruit
citrus fruits. Citrus species are the most quality varieties for optimal growing (Yılmaz
important fruit groups for Turkey in view of et al., 2013).
production and export quantities. Orange The fruit quality of the citrus species is affected
(Citrus sinensis) is the first species among by cultivars or types, rootstocks, ecology and
Citrus with high level of production. Tropical cultural practices (Ozcan and Ulubelde, 1984;
and semitropical citrus fruits are more Economides and Gregoriou, 1993; Castle 1995;
concentrated in subtropical regions (Davies and Tuzcu et al., 1999). Similarly Hodgson (1967)
Albrigo, 2005). Located in the Mediterranean emphasized that fruit quality is influenced by
basin, Turkey has quite favorable ecological three main factors as climate, rootstock and
conditions for production of quality edible nutrition. Moreover, climate is the most impor-
citrus (Tuzcu, 1998). China, Brazil and India tant parameter affecting fruit quality and size.
are the top countries in world citrus production In this study, the pomological characteristics of
(FAO, 2017). Turkey is the 9th country with some sweet orange varieties have been
4,293,007 tons among the major citrus evaluated in the Antalya region, which has an
producers in 2016 (TUIK, 2017). The citrus important potential for citrus growing.
fruits are generally the first in the total export
of fresh fruits and vegetables in Turkey (AKİB, MATERIALS AND METHODS
2017).
The present research was conducted on citrus
Although Turkey is no homeland of citrus, a lot
germplasm collection of Bati Akdeniz
of varieties have been brought from different
Agricultural Research Institute (BATEM) in
countries has led to the emergence of an
Antalya (Turkey) during 2015 and 2016. Six
important gene source. The ecological
orange varieties (Belladonna, Biondo, Biondo
conditions of the Mediterranean and Aegean
Riccio, Calabrese, Mediterranean Sweet and
regions in Turkey allow for citrus cultivation
Parson Brown), budded on the sour orange
successfully. In his way, Turkey has been the
‘Yerli’ (Citrus aurantium L. var. 'Yerli') in
135
1938, were used as plant material. The terms of both yield and quality even in
experimental plot located at 36° 52’ 29.9’’ N advanced ages under appropriate cultural
and 30° 43’ 28.5’’ E latitude, the altitude is conditions (data were not shown here).
about 37 m. Sweet orange orchard had a typical Results of average of some pomological
Mediterranean climate, and soil had alkaline characteristics of six orange varieties for last
reaction, calcareous, unsalted, and rich in two consecutive years are shown in Table 1 and
phosphorus. Table 2.
The fruits were randomly harvested from all Fruit quality is an occurrence that expresses
sides of different 4 trees for each variety in many different parametrizes. Each fruit has
middle-January on both trial years. 20 healthy specific fruit quality factors although there are
fruits were used for pomological analyzes. The similarities among others. Janick and Moore
fruit quality parameters of orange varieties (1975) set these factors as: fresh size, shape,
were assessed according to (Ozsan and colour, peel, juice soluble solid, solid: acid
Bahçecioglu, 1970). ratio, flavor, ease of peeling, seed content and
Different physical and chemical parameters i.e. juice content.
fruit weight, fruit height, fruit diameter, rind It is well known fruit size is very important
thickness, segment number, seed number, juice especially consuming as fresh fruit. The
content, total soluble solids (TSS), total acidity variance analysis showed that fruit weight of
(TA) and ascorbic acid were evaluated. Fruit orange varieties was significant (P≤0.05). The
dimensions and peel thickness were measured highest fruit weight was the obtained from
with a digital caliper (0.01 mm sensitivity). The Mediterranean Sweet (187.54 g) while the
fruits, which had almost same color and lowest one was Biondo Riccio (151.71 g).
dimensions, were weighed by 0.01 g sensitivity Tuzcu (1990) reported these values of different
balance (Shimadzu) and their juice extracted. orange varieties grown in Turkey as; 191.16 g
The juice retrieved was weighed and calculated (Yafa), 160.81 g (Valencia), 160.99 g
the fruit content percentage. Segment number (Hamlin), 198.91 g (Finike Yerli), 162.36 g
and seed number were counted. Total soluble (Alanya Dilimli), 165.97 g (Kozan Yerli),
solid (TSS in oBrix) was measured using hand 164.06 g (Dörtyol Yerli). The weight of
refractometer (N.O.W TokyoͲ̱͵ʹ). Titratable different Valencia clones in Antalya changed
acidity (TS) was determined with AOAC between to 228.71 g (VAA 75) and 214.82 g
method (NaOH 0.1 N to pH 8.1) using a titrator (VAA 59) (Tuncay, 2005). Fruit weight was
and expressed as grams of anhydrous citric determined by Altan (1995) as 205 g (Hamlin),
acid/100 ml fruit juice. Maturity index 164 g (Magnum Bonum), 148 g (Dörtyol
(TSS/TA) was calculated. pH of fruit juice was Yerli), 196 g (Kozan Yerli) and 157 g
measured by digital pH meter (WTW Ino Lab). (Valencia) in Çukurova Region. Khan (2015)
The ascorbic acid content was determined as stated that fruit weight was between 189.75 g
reported earlier by using method of Horwitz (Pineapple) to 140.50 g (Robel) in Pakistan.
(1975) and expressed mg/100 ml.) Even if fruit weights of our results are different,
The experimental design was performed they are included in the boundary of others.
according to a randomized design with 4 The current differences can be attributed to
replications, each replication consisting of one differences of tree age and ecology.
tree. Data analysis was done by analysis of The highest average fruit height for last two
SAS statistical software and means were years was taken from Calabrese (70.99 mm)
separated by using LSD test at (P≤0.05). and Mediterranean Sweet (70.02 mm).
Average data of last 2 years were presented in Statistically significant differences were
this present study. obtained between orange varieties expressed in
Table 1. Similarly, fruit diameter of trial
RESULTS AND DISCUSSIONS oranges has significant difference (P≤0.05)
according to variance analysis.
It has been observed that the sweet orange As seen in Table 1, Mediterranean Sweet had
varieties, budded on C. aurantium L., are able the widest fruits diameter with 71.81 mm,
to maintain their commercial significance in while Biondo Riccio had the small one with
136
67.09 mm. Tuncay (2005) stated that the fruit it does not affected to rootstocks, climatic
length/width of Valencia and Yafa clones conditions and etc. This notification is confirm
grown in Antalya were the 77.14 mm (VAA to our results, which there was no significant
72)/ 76.13 (VAA 70) mm and 91.21 mm/ difference of segment number among trial
(YAA 46) 80.92 mm (YAA 45). varieties (Table 1).
When compared to the rind thickness of the Fruit quality and its appearance are very
fruits there was no significant differences, even important from a consumer’s viewpoint.
though the thin-rind fruit obtained from cv. Especially seedless fruit are desired quality
Biondo (3.66 mm), Mediterranean Sweet (4.07 characteristics for orange citrus. In this study,
mm) had the thicker-rind. All these reports Calabrese, Biondo, Biondo Riccio and
indicated that fruit dimensions and rind Mediterranean Sweet oranges were defined as
thickness can vary to ecology. commercial seedless varieties in Antalya (south
Tuzcu et al. (1999) have stated that segment Turkey).
number is an important genetic factor and also
Table 1. Main fruit characteristics of orange varieties (mean of two years)

Fruit weight Fruit height Fruit width Rind thickness Segment Seed
Variety
(g) (mm) (mm) (mm) number number
Belladonna 175.18±21.77 ab 66.38±1.50b 70.99±1.97a 3.80±0.32 9.37±0.44 c 6.84±1.56 cd
Biondo 162.36±15.55 bc 65.67±1.10 bc 67.68±2.84b 3.66±0.35 9.45±0.24 c 4.55±1.56 ab
Biondo Riccio 151.71±30.64 c 63.54±4.81 c 67.09±4.14b 3.92±0.13 9.36±0.36 c 5.55±1.06 bc
Calabrese 173.43±14.89 ac 70.99±1.64a 69.53±2.62ab 3.97±0.22 9.87±0.27 b 3.07±0.96 a
Mediterranean
187.54±10.32 a 70.02±2.50 a 71.81±1.37a 4.07±0.17 10.33±0.33 a 5.16±0.84 bc
Sweet
Parson Brown 160.24±15.38 bc 63.07±1.03 c 68.99±2.23 ab 3.97±0.24 9.91±0.36 b 8.09±3.17 d
LSD (0.05) 22.006 2.8231 3.0545 NS 0.3876 1.964
*Mean separation within columns by LSD multiple range test P≤0.05.
Fruit quality, is eventually a matter of significant difference among orange cultivars.

consumer preference, can measure as physical Juice content was expressed as different ratio in
traits and chemical composition. In current a lot of research established different ecology
study, some chemical properties are evaluated (Tuzcu, 1990; Tuzcu et al., 1993; Yılmaz et al.,
and shown in Table 2. Juice content changed 2013). Total soluble solid (TSS), juice acidity
between 59.78% (Calabrese) 56.91% (TA) and TTS/TA are abundant quality
(Mediterranean Sweet). There was no parameters in citrus.
Table 2. Main fruit quality properties of orange varieties (mean of two years)
Fruit juice content Total Soluble solids Soluble solids/ Ascorbic acid
Variety Acidity (%) pH
(%) (%) Acidity mg/100ml
Belladonna 56.94±2.09 8.33±0.57 bc 1.44±0.12 5.90±0.95 3.38±0.43 54.86±2.04bc
Biondo 59.70±2.37 9.30±0.95 b 1.70±0.31 5.81±1.65 3.31±0.37 49.60±1.76de
Biondo Riccio 57.22±3.04 8.85±1.20 bc 1.71±0.41 5.72±2.05 3.27±0.33 57.10±2.58b
Calabrese 59.78±3.62 7.48±1.44 c 1.51±0.11 5.04±1.25 3.07±0.21 46.71±1.40e
Mediterranean Sweet 56.91±2.33 8.83±1.82 bc 1.49±0.23 6.68±2.51 3.08±0.13 52.44±6.21cd
Parson Brown 56.82±1.15 11.00±1.94a 1.58±0.20 7.26±2.11 3.32±0.51 61.20±2.86a
LSD (0.05) NS 1.6049 NS NS NS 3.6987
*Mean separation within columns by LSD multiple range test P≤0.05.
Parson Brown had the highest with 11% TTS trial varieties were no significant. Fruit quality
but Calabrese had the lowest (7.48%), and is foremost an inherent scion cultivar trait. It
difference among orange varieties was can be modified but not radically changed
significant (P≤0.05). Our TTS results are without genetic manipulation (Castle, 1995).
almost similar to reports of Khan (2015) and Among the secondary factors, climate is widely
Tuzcu (1990). TA, TTS/TA and pH values of recognised as a major fruit quality factor
137
judging by the prominence of climatic commercial significance in terms of both yield
adaptation on plant breeders' lists of objectives and quality even in advanced ages under
(Hodgson, 1967; Janick and Moore 1975). In appropriate cultural conditions.
citrus, rootstocks have many scion interactive
effects; the principal internal factors are juice ACKNOWLEDGEMENTS
content and colour, and soluble solids, acid
concentrations and their ratio (Wutscher, The authors thank The Scientific Research
1988). Projects Coordination Unit of Süleyman
Citrus fruits have been consumed for good Demirel University of Turkey for their
sources of antioxidant. Stuetz et al. (2010) financial support (Project No: 4641-YL1-16)
stated that they were acknowledged as a good
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reticulata Blanco cv. ʻSainampuengʼ) from Northern Tuzcu O., Yıldırım B., Düzenoglu S., Bahçeci İ., 1999.
Thailand J. Agric. Food Chem, 58: 6069-6074 6 Değişik turunçgil anaçlarının Washington Navel ve
Tounsi MS., Wannes W.A., Ouerghemmi I., Jegham S., Moro kan portakal çeşitlerinin meyve verim ve
Njima Y.B., Hamdaoui G., Zemni H., Marzouka B., kalitesi üzerine etkileri. Turkish Journal of
2011. Juice components and antioxidant capacity of Agriculture and Forestry, 23: 213-222.
four Tunisian citrus varieties. J. Sci. Food Agric, 91: Wutscher H.K., Bistline F.W., 1988. Rootstock
142-151. influences juice color of 'Hamlin' orange. Hort
TUİK., 2017. Statistical database. Science 23 (4): 724-725.
https://biruni.tuik.gov.tr/bitkiselapp/bitkisel.zul. Yılmaz B., İncesu M., Çimen B., Yeşiloglu T., Pamuk
Accessed:12 May 2017. S., 2013. Bazı göbekli portakal çeşitleri ile türkiye’de
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Turunçgillerde aşıgözü seleksiyon- sertifikasyonu ve adana ekolojik koşullarında verim ve kalite
özelliklerinin belirlenmesi. TABAD, 6 (2): 05-11.
139
140
POLLEN PERFORMANCES OF NATURALLY GROWN BLACKBERRIES

IN ISPARTA - TURKEY
Sultan Filiz Güçlü1, Ayşe Gül Sarıkaya1, Fatma Koyuncu2

1
Süleyman Demirel University, Atabey Vocational School, Isparta, Turkey
2
Süleyman Demirel University, Faculty of Agriculture, Horticultural Science, 32260, Isparta,
Turkey
Abstract
Fertilization biology is one of the most important criteria used to identify as candidate genotype and variety. In this
research pollens of 6 wild-grown blackberry shrubs, preselected from Isparta - Turkey, were used. In order to
determine the pollen performance which is the basis of the studies on fertilization biology, the amount of pollen
production, pollen viability degree and the morphological homogeneity rate were determined. In addition, temperature
experiments (150C, 200C and 250C) were performed to determine the optimum pollen germination condition. However,
the effects of some growth regulators and mineral substances (GA3, KNO3, BA) on pollen germination and tube growth
were investigated. N6 had the highest values in terms of anther number (71.2) average number of pollens per anther
(13151.7), number of pollen per flower (414245.7), and morphological homogeneity (95.8%). 200C was found optimum
temperature for pollen germination and tube growth. Gibberellic acid and Potassium nitrate were determined as
promoter while benzyl adenine had inhibitory effect on pollen germination and tube growth. As a result of fertilization
biology studies N6 can be thought promising, terms of pollen performance.
Key words: Rubus canescens, pollen tube growth, TTC, KNO3.
INTRODUCTION to shortly tomentose, armed like the turions.

Sepals are ovate-oblong, acute, pubescent
Turkey is located in the region of the tomentose, reflexed after flowering. Petals are
intersection of three phytogeography regions. white (drying pale yellowish), obovate-oblong
While Turkey flora has approximately 11,466 (rarely broader), 5-8 (-10) mm. Fruits are about
plant taxon, whole Europe continent has 1 cm in size and red purple color. Drupelets are
approximately 12,000 plant taxon. Rosaceae black, small and numerous, glabrous (Davis,
family has some are thorny and trailing that are 1972).
rarely grassy, bush and tree exist. This family is It is light-half-shadow plant. It is resistant to
represented by 115 genus and approximately cold and hot. It likes moisture but can also
3200 species in the world. Rosaceae family grow in dry places. It is used of forceps,
members are represented by 35 genus and 318 diuretic and diabetes in folk medicine. The
species in Turkey. Rubus taxon is an important liquid obtained from the fruits is used as a
genus of Rosaceae family. mouthwash against inflammations of the tonsils
It has about 16 species in Turkey (Güner, (Durmuşkahya, 2006).
2012). Rubus canescens DC. var. canescens is Fertilization success in plants is the result of
low, usually trailing shrub. The red purplish processes that take place during the progamic
body is angular and often prickly. Thorns are phase (Thompson, 2004; Güçlü and Koyuncu,
short, flat and sickle-shaped. Leaves are ternate 2017). Pollen germination and pollen tube
or sometimes pedate with 3 or 5 leaflets, growth are important components of
discolorous; leaflets tomentellous, dull green or fertilization success in fruit trees (Janıck and
greyish green, canescent-tomentose with Moore, 1996; Tosun and Koyuncu, 2007).
stellate hairs below, bidentate, lateral ones The first condition of formation of seed and
subsessile, terminal one petiolulate, cuneate- fruit is developing healthy male and female
obovate to rhombic; stipules linear. Flowering organs of the flower and cells, except for an
shoots are erect, 15-40 cm, angled, pubescent apparent partenocarpy of some cultivars.
141
Pollen performance, which includes pollen The final percentage of morphological homo-
germination, pollen tube growth rate and pollen geneity (MH) was defined as:
competition, is an important component of ܰ‫ ݏ‬െ ܰܽ
fertilization success in seed-producing plants. ‫ ܪܯ‬ൌ ‫ͲͲͳݔ‬
Pollen performance is clearly affected by the ‫ܣ‬
genotype of the pollen (Acar and Kakani, 2010; Ns: Number of shaped pollen per area;
Hedly et al., 2004). Na: Number of aborted pollen per area;
Pollen-pistil interactions and environmental A: Total number of pollen area.
factors also affect pollen performance (Dafni
and Firmage, 2000). Temperature is one of the Pollen viability was determined by using TTC
most important environmental factors for (2, 3, 5-triphenyl tetrazolium chloride) staining
pollen germination, fruit set and seed set. test. A few drops of 1% TTC (0.2 g. TTC and
Temperature has been shown to affect the 12 g. sucrose were dissolved in 20 ml distilled
chemical composition of pollen, pollen water) were dropped by Pasteur pipettes on
viability, pollen tube growth as well (Johanson microscope slides and pollen were shaked with
and Stephanson, 1998). a slim brush (each brush used only one plant
Pollen germination and pollen tube growth are type) covered with a coverslip of used four
important research materials for morphological, microscope slides with three replication were
physiological, biotechnological, ecological, counted after 2 hours.
evolutional, biochemical, systematic and To determine the pollen viability, pollens of
molecular studies. Additionally testing pollen each cultivar (of four different areas) were
performance could be helpful for a fruit observed onto two slides under a light
cultivation of genetic progeny for breeding microscope (×100 magnification). The stained
purpose, and especially for selecting which pollen was considered as ‘viable’ in the test.
cultivars should be used by researchers and The ‘agar in plate’ method was used to
growers. establish pollen germination and pollen tube
For this purpose we tried to observe the pollen growth (Koyuncu and Tosun, 2009).
performances of 6 wild-grown blackberries. Pollen tube long at least as its diameter was
considered to be ‘germinated’. For temperature
MATERIALS AND METHODS experiments; pollen grains were sowed in the
medium containing 15% sucrose+0.5% agar-
Pollens were collected from flowers of 6 wild- agar+5 ppm (H3BO3) boric acid at 150C, 200C
grown blackberry shrubs (N1, N2, N3, N4, N5, and 250C in the dark.
N6) at balloon stage. For the growth regulators and mineral
The 50 flowers at balloon stage were picked substances experiments GA3, KNO3 and BA
up. The flowers were transferred to the were added at the determined doses as a
laboratory immediately. Anthers were removed preliminary experiment (50 ppm GA3, 50 ppm
and placed into the dark-colored bottle to KNO3, and 50 ppm BA) (Tosun and Koyuncu,
promote dehiscence at room temperature. For 2007b).
the pollen performance, pollen production An ocular micrometer was used to measure
capacity, morphological homogeneity, pollen pollen tube length, under a light microscope, at
viability rates were investigated. a 40x magnification. Four Petri dishes were
In addition, optimum germination temperature used for germination and pollen tube growth
and optimum germination medium consist of experiments. For each assay, 2 mL of medium
some growth regulators and mineral substances was placed into Petri dishes. Counts were made
were determined. The pollen production from 4 different microscope fields (100-150
capacity and morphological homogeneity pollen grains per field for each Petri) (Hedly et
percentages of pollens were assessed with the al., 2004; Koyuncu, 2006).
hemocytometer (Marienfeld, Germany) slide Statistical analysis was conducted using
(Eti, 1990). Imperfectly shaped pollen grains Duncan’s multiple range test within the general
were considered as aborted pollen. linear model procedure of SPSS 16.0.
142
RESULTS AND DISCUSSIONS Pollen germination and pollen tube growth
tests
Pollen production capacity and pollen viability Three constant temperature regimes (150C,
Pollen production amount and viability results 200C and 250C) were evaluated on the pollen
were shown in Table 1. The difference in the germination and expressed as the percentage of
amount of pollen production of genotypes was germinated pollen (Table 2). As seen as Table
statistically significant (p<0.05). The highest 2, the effects of different constant temperature
results were taken from N6, in terms of on pollen germination were statistically
numbers of anthers in a flower (71.2), mean significant (p<0.05). N6 is the best genotype
pollen number in an anther (13,151.74) and with highest value (52.25) in terms of pollen
pollen number in a flower (414,245.07). germination rate, at all temperature. This is
However, the difference between pollen followed by N1 (32.40) and N5 (23.39),
viability and morphological homogeneity were respectively. The lowest pollen germination
not statistically significant (p<0.05). The pollen rate was obtained from N3 (7.29). 200C was
viability rate ranged from 79.88% (N2) to determined as the optimum germination
83.22% (N6). Morphological homogeneity temperature for all genotypes. When the
rates were found upper than 92%. Türemiş and temperature rose up from 150C to 200C, the
Derin (1999), reported that pollen viability pollen germination rate also increased
levels varied from 79.75% to 91.94% some dramatically, but when it reached 250C,
blackberry cultivars. Koyuncu (2006) studied germination began to fall down. This shows us
strawberry pollens using TTC viability test and of high temperatures had the negative effect on
reported that pollen viability ratios reached to the pollen germination.
82% for cvs. ʻAllstarʼ and ʻElviraʼ and 86.5%
for cv. ʻChandlerʼ. Koyuncu and Tosun (2009) Table 2. In vitro pollen germination (%) of blackberries
used TTC, FDA and IKI stain tests for the pollen at different temperatures after 24 hours
incubation, an a medium containing 15% sucrose + 0.5%
sweet cherry cultivars. They reported that the agar-agar + 5 ppm (H3BO3)
pollen viability differed according to stain
methods and cultivars. The viability and Incubation temperature (oC)
15 20 25 Mean
morphological homogeneity related to pollen
N1 9 48.21 40 32.40bx
quality. They are the most important properties
N2 8.56 12.9 12 11.15d
in fruit trees. These properties are useful for
N3 2.96 10.5 8.41 7.29d
plant breeders, geneticists and growers.
N4 5.78 13.24 10.67 9.90d
However, an easy method for determining
N5 7.25 33.59 29.34 23.39c
pollen viability is required to increase the N6 16.64 75.87 64.23 52.25a
efficiency of the breeding program and the Mean 8.37cy 32.39a 27.44b
selection of a suitable pollinizer while the x
Values within a column followed by different letters are
orchard is being established (Ercisli, 2007). significantly different (p<0.05).
y
Values within same row followed by different letters are
Table 1. Pollen production capacity, morphological significantly different (p<0.05).
homogeneity and pollen viability test
n m pn pv (%) MH (%)
The in vitro elongation of pollen tubes was
* x x
affected by incubation temperature (Table 3).
N1 67.4b 12919.14b 391742.11b 83.10 95.12
Responses of tested cultivars to different
N2 65.5b 12431.10b 371130.07b 79.88 93.5
temperatures were statistically significant
N3 66.3b 12782.19b 383435.14b 81.16 92.2
(p<0.05). As seen as Table 3, the longest pollen
N4 66.9b 12830.71b 392424.16b 80.23 92.9
tubes for all varieties were measured at 200C
N5 65.8b 12754.09b 374126.09b 81.63 94.0
when the shortest ones at 150C. The most
N6 71.2a 13151.74a 414245.07a 83.22 95.8
augmentation pollen tube elongation was
n: numbers of anthers in a flower; m: mean pollen
obtained from N6 at 15 to 200C (95.88 µm-
number in an anther, pn: pollen number in a flower; pv:
pollen viability, MH : morphological homogeneity. 154.36 µm). Pollen tube length of N6 was
above the average (Figure 1).
*Values within a column followed by different letters are
significantly different (p<0.05).xThe difference between
the values is not statistically significant. SUB TABEL 1!
143
Table 3. The effect of incubation temperature on pollen varies among species and cultivars of the same
tube growth (µm) after 24 hours species (Mert, 2009; Kuroki et al., 2017).
Incubation temperature (oC) The effects of chemicals on pollen germination
o
15 C 20 oC 25 oC Mean and tube growth were statistically different,
N1 67.86 89.74 82.51 80.04dx (p<0.05). Potassium nitrate and gibberellic acid
N2 91.26 101.36 95.9 96.17b were determined as promoter while benzyl
N3 72.25 98.25 84.63 85.04d
adenine effects as inhibitory pollen germination
and tube growth (Tables 4, 5). Pollen
N4 84.26 97.65 92.54 91.48c
germination rate decreased to 24.82 in BA
N5 92.33 106.67 99.51 99.50b
supplemented medium and increased to 30.72%
N6 95.88 154.36 148.69 132.98a in GA3 added medium by 41.16%. The study
Mean 83.97cy 108.01a 100.63b which was conducted at blackberries it is
x
Values within a column followed by different letters are reported that the effects of minerals on pollen
significantly different (p<0.05).
y
germination were found to vary according to
significantly different (p<0.05). cultivars and doses (Türemiş and Derin, 1999).
Kumar et al. (2016) found GA3 as a promoter
for ornamental tropical tree species pollens.
Table 4. The effect of different plant growth regulator on

pollen germination (%)
Control BA KNO3 GA3 Mean
N1 48.21 36.2 52.7 55.3 48.10bx
N2 12.9 8.2 14.71 19.34 13.78d
N3 10.5 7.7 18.6 24.5 15.32d
N4 13.24 10.2 18.22 26.4 17.01d
N5 33.59 27.7 39.79 45.61 36.67c
Figure 1. Pollen tube elongation N6 65.87 58.9 68.71 75.8 67.32a
at different incubation temperature Mean 30.72cy 24.82d 35.46b 41.16a
Temperature is among the most important

Table 5. The effect of different plant regulators on pollen
environmental factors affecting plant tube length (µm) after 24 hours
reproductive processes such as pollen
Control BA KNO3 GA3 Mean
germination pollen tube growth and fruit set
(Kakani et al., 2005). Temperature ranges and N1 98.22 94.65 114.26 128.11 108.81cx
optimum temperature values for pollen N2 103.3 82.33 109.55 117.09 103.06c
germination and tube growth were studied for N3 94.68 103.26 113.6 120.39 107.98c
different fruit species, including strawberry N4 145.63 127.58 139.41 154.69 141.82b
(Koyuncu, 2006), pears (Vasilakasis and N5 130.65 107.89 156.86 165.99 140.34b
Porlingis, 1985), cherries (Tosun and Koyuncu, N6 151.36 144.85 161.48 174.25 157.98a
2007a), pistachio species (Acar and Kakani, Mean 120.64cy 110.09d 132.52b 143.42a
2010), pomegranate (Gökbayrak and Engin, x
Values within a column followed by different letters are
2018) The optimum temperature required for significantly different (p<0.05).
pollen germination and tube growth was about y
200C for apricot, cherry and sour cherry significantly different (p<0.05).
(Austin et al., 1998; Koyuncu and Tosun,
Pollen germination regulated by water, amino
2009). These results parallel to our findings.
acids, sugars, calcium and growth regulators
Another study which was conducted at apricots
such as gibberellins, auxins and kinetin.
as our study, pollen germination and tube
Gibberellins have been traced in developing
growth above 25°C reduced (Egea et al., 1992).
pollen grain after anthesis (Singh et al., 2002).
Low or high temperatures have negative effects
Our results have been supported these findings.
on pollen germination and tube growth. The
Pollen performance criteria (pollen vitality,
optimum temperature for pollen germination
144
germination and pollen tube growth rate) are Egea J., Burgos L., Zoroa N., Egea L., 1992. Influence of
critical for discharging male gametes in the Temperature on the In Vıtro Germination of Pollen of
Apricot (Prunus armeniaca L.,). J. Hort. Sci. 67, 247-
embryo sac and are a prerequisite for 250.
fertilization and fruit set. In vitro germination Ercisli S., 2007. Determination of pollen viability and in
studies are powerful tools for genetic, vitro pollen germination of Rosa dumalis and Rosa
physiological, biochemical and cytochemical villosa. Bangladesh J. Botany, 36: 185-187.
studies for a wide range of plant species Eti S., 1990. Cicek tozu miktarini belirlemede kullanilan
pratik bir yontem. Journal of Agricultural Faculty,
belonging to different families (Radıčevıć et Cukurova University, 5: 49–58.
al., 2013). These studies are also a good Fotirić Akšić M., Cerović R., Rakonjac V., Bakić I.,
predictor of in vivo pollen behavior but only for Čolić S., Meland M., 2017. Vitality And In Vitro
autotrophic phase of pollen growth where the Pollen Germination Of Different 'Oblačinska' Sour
initial steps of pollen germination and pollen Cherry Clones. Genetika (0534-0012), 49 (3), 791-
800. Doi:10.2298/Gensr1703791a.
tube growth are independent of style nutrients, Gökbayrak Z., Engin H., 2018. Effects of Foliar-Applied
sugars and plant growth regulators. They help Brassinosteroid on Viability and In Vitro
with selections for breeding programmers, in Germination of Pollen Collected from Bisexual and
vitro assessments can also help to predict Functional Male Flowers of Pomegranate.
possible problems of sterility of that particular International Journal of Fruit Science, 1-5.
Güclü S.F., Koyuncu F., 2017. Effects Of Relative
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Akšić et al., 2017) Growth In Sweet Cherries (Prunus avium L.)
Scientific Papers. Series B, Horticulture. Vol. LXI,
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Güclü S.F., Koyuncu F., 2017. An In Vitro Study of
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determined as promoter while benzyl adenine sweet cherry, Prunus avium (Rosaceae). Am J Bot 91
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146
EFFECT OF POSTHARVEST OXALIC ACID TREATMENT

ON COLD STORAGE OF APRICOT CV. ʻAPRIKOZʼ
Mehmet Ali KOYUNCU1, Tuba SECMEN2, Cemile Ebru ONURSAL2, Derya ERBAS1,
Atakan GUNEYLI3, Seda SEVINC UZUMCU3, Fatma KOYUNCU1
1
Süleyman Demirel University, Faculty of Agriculture, Department of Horticulture, Isparta, Turkey
2
Batı Akdeniz Agrıcultural Research Institute, Antalya, Turkey
3
Fruit Research Institute, Isparta, Turkey
Abstract
In this study, effects of postharvest oxalic acid (OA) treatments on storage life and quality of apricot cv. ʻAprikozʼ were
investigated. Fruit were harvested at optimum stage (firm ripe stage) and transported to the postharvest physiology
laboratory, immediately. Apricots were immersed in different doses (0, 1, 2, 4 and 8 mM) of OA solution+ Tween 20 for
10 minutes. After treatments, fruit were held at room conditions for drying during 30 minutes. Dried fruit were placed
in modified atmosphere packages (MAP) and stored for 40 days at 0°C and 90±5% relative humidity. The weight loss,
total soluble solids content, titratable acidity, fruit flesh firmness, fruit skin color, respiration rate and gas composition
of MAP were determined at the beginning of the storage and 10-day intervals during the cold storage period. As a
result, all doses of OA gave better results than control group in terms of some quality parameters. Especially, 1 mM
dose of OA was the most effective treatment for decreasing weight loss and maintaining fruits firmness. The results
suggest that OA has the potential to extend the storage life of apricot by delaying quality loss.
Key words: apricot, oxalic acid, modified atmosphere, cold storage.
INTRODUCTION plays an important role in physiological

functions such as regulating stress responses,
Apricots are one of the most popular fruit in resistance against disease (Wu et al., 2011). In
both domestic and international markets owing addition, postharvest treatments with OA were
to its delicious taste and aroma (Özdoğru et al., effective in delaying some processes of
2015). Apricots have sufficient amounts of climacteric fruit through an inhibition of
sucrose, glucose, fructose and high antioxidant ethylene biosynthesis (Martinez-Espla et al.,
components (lycopene, β-carotene, vitamins A 2014). Another beneficial effect of pre-storage
and E) and minerals (K, P, and Mg). OA applications is reduced chilling injury in
Consumption of apricots plays an important pomegranate and mango in connection with an
role in preventing diseases and maintaining enhance in antioxidant capacity (Li et al.,
healthy life (Muradoğlu et al., 2011). Orchard 2014). In previous studies, it was reported that
conditions, developmental stage at harvest, and both pre- and postharvest OA treatments
other postharvest factors such as chilling injury, extended storage life, and maintained fruit
high temperature and mechanical damage quality during the storage period. Zheng et al.
influence fruit quality. Fruit quality in apricot is (2007a; 2007b) stated that OA treatments in
an important factor that affects consumers’ mango and peach fruit were effective in
perception. Apricots as climacteric stone fruit extending the postharvest life. Razavi and
have a limited postharvest life (Ezzat et al., Hajilou (2016) treated sweet cherries with OA
2017). The main factor limiting the postharvest and stored them for 20 days under cold
life of apricot fruit are very rapid maturation temperature, and reported that OA delayed the
process due to the high rate of respiration after postharvest ripening. In literature, it can be
harvest (Abd El Wahab, 2015; Jing et al., found some researches related to OA but there
2018). Oxalic acid (OA), as a final metabolite are not enough studies about the effect of OA
product in plants (Martinez-Espla et al., 2014), on the postharvest life and quality of apricot
147
during cold storage. The aim of this study was colorimeter over 15 fruit in each replicate. The
to investigate the effect of postharvest OA values were expressed by the CIE L*
treatment on cold storage of apricot cv. (brightness-darkness), a* (+a*: red, −a*: green)
ʻAprikozʼ. and b* (+b*: yellow, −b*: blue) system.
Respiration rate and ethylene production were
MATERIALS AND METHODS measured in 600-700 g of fruit samples for
each replicate. Fruit were weighed and placed
Harvest and postharvest treatments in 2 L airtight jars for 2 h at 20ºC. Then gas
The fruit of apricot cv. ʻAprikozʼ were sample was taken from jars and injected into
harvested at commercial harvest stage gas chromatographs. Results were expressed as
(yellowish-green and firm ripe), and μL/kg.h for ethylene production and mL
transported to the postharvest laboratory, CO2/kg h for respiration rate. Gas concentration
immediately by a frigofric car. Fruit were (O2 and CO2) in the packages was measured by
selected for uniformity (color, size and shape) Gaspace 2 (Gas Headspace analyzer, Systech
and any blemished/diseased fruit were Instruments) and expressed as percentage.
discarded. Selected apricots were randomly External appearance was rated on a hedonic
divided into five groups. First group (control) scale of 1-9 (1-3: unmarketable, 5: marketable,
apricots were dipped into distilled water + 7: good, 9: very good), taste was rated on a
0.01% Tween-20 (a surfactant) for 10 min. The hedonic scale of 1-5 (1: very bad, 2: bad, 3:
other four group apricots were dipped into in medium, 4: good, 5: very good) and internal
different doses (1, 2, 4 and 8 mM) of OA browning was rated on a hedonic scale of 0-4
solution + 0.01% Tween 20 for 10 minutes. (0: healthy, 1: 1-10%, 2: 11-33%, 3: 33-66%, 4:
After dipping treatments, fruit were held at 66-100%). The experiment was set up
room conditions for drying during 30 minutes. according to the factorial randomized design
Dried fruit were placed in modified atmosphere with 3 replications (40 fruit per replication).
packages (MAP) (25 μm low-density Data were subjected to analysis of variance
polyethylene) and stored for 40 days at 0°C and (ANOVA, JMP7), means were separated by
90±5% relative humidity. All analyses were means of Tukey test (P<0.05).
performed at harvest date and 10 days intervals
during cold storage. RESULTS AND DISCUSSIONS
Chemical and physical analysis
Weight loss of apricots was measured over 15 Weight loss. Weight losses of apricots treated
fruit in each replicate and expressed as the with different doses of OA, during the cold
percentage of loss of weight with respect to the storage, was given in the Table 1. Weight loss,
initial weight. Weight loss was determined by one of the most important factors limiting the
the formula; Weight loss = [(First weight - Last storage life of the products, has increased
weight)/First weight] × 100. Fruit flesh continuously during storage. But this increase
firmness was measured over 15 fruit in each was found to be lower in OA treatments
replicate. Fruit flesh firmness (a small slice of (except for 4 mM) than in control groups. The
fruit skin was removed from each side of a weight loss of the apricots treated with dose of
fruit) was determined using a digital texture 1 mM OA (1.36 %) was significantly delayed
machine and measured via compression using a compared to control and other doses at the end
50 N load cell and a stainless steel, 5.1 mm of cold storage (Table 1.) Fruit lost their weight
diameter. The results were expressed as mainly due to respiration and transpiration
Newton (N). Total soluble solid (TSS) content through skin and various metabolic activities.
was measured using a digital refractometer The positive effects of OA on weight loss of
(Atago Pocket PAL-1) and expressed as fruit might be due to slowed metabolic process
percentage. Titratable acidity (TA) was and decreased respiratory rate (Razzaq et al.,
determined by a digital pH meter (Hanna 2015). According to some researches which are
Instruments HI 9231) and titrimeter (Digitrat, parallel to our results, OA-treated fruit
Isolab), and expressed as percentage. Fruit skin exhibited reduced weight loss compared with
color was determined using a Minolta CR-300 control (Sayyari et al., 2010; Razzaq et al.,
148
2015). Sayyari et al., 2010 reported that pre- the postharvest life of fruit. Correspondingly
storage OA treatments reduced the weight loss Wu et al. (2011) stated that the application of
in pomegranate fruit stored at 2oC for 84 days. OA delayed softening of plum fruit. They
Also, Razzaq et al. (2015) reported that OA suggested that the inhibition of softening was
treatments reduced weight loss in mango fruit. associated with decreased polygalacturonase
Table 1. Weight loss (%) of ʻAprikozʼ apricots treated
(PG) and pectin methyl esterase (PME)
with OA during MAP storage at 0°C activities; that is, the retardation of pectin
solubilization/degradation (Razavi and Hajilou,
Treatment 10 d 20 d 30 d 40 d Means
Control 0.55 1.34 1.94 2.58 1.59 ab*
2016).
1 mM 0.56 1.17 1.53 2.23 1.36 c Total soluble solid (TSS) content and
2 mM 0.64 1.43 1.55 2.53 1.53 ab titratable acidity (TA). The effects of OA
4 mM 0.60 1.50 1.93 2.60 1.65 a
8 mM 0.51 1.34 1.57 2.45 1.46 bc
treatments on TSS content and TA were
Means 0.57 d 1.35 c 1.70 b 2.47 a statistically significant (P<0.05). The TSS and
*Means followed by the same letter in the same row and TA decreased in all treatments with increasing
column are not statistically significant (P<0.05); d: days.
storage period. The TSS of fruit treated with
Fruit flesh firmness. The OA treatments had 1.0 and 8.0 mM doses were significantly higher
positive effects on fruit firmness. The flesh than those of control. The TSS contents varied
firmness in treated fruit was maintained between 10.50% and 9.57% at the end of 40
compared with the control, thus fruit softening days (Table 3). The TA contents of fruit
rate was delayed by OA during storage. The gradually decreased over the storage period
highest firmness value (34.43 N) was regardless of treatments. The highest TA value
determined in 1 mM OA treatment, whereas the was obtained from 1 mM dose of OA varying
lowest (30.71 N) was from control fruit (Table in the range of 1.10-0.92 throughout storage
2). There are similar results obtained by other period. Zheng et al. (2007a) reported that OA
researchers. Studies on peach (Razavi and treatments in mango, a climacteric fruit,
Hajilou, 2016; Zheng et al., 2007b), mango increased TSS while TA was decreased.
(Zheng et al., 2007a) and plum (Wu et al., Adverse results of Zheng et al. (2007a; 2007b)
2011) showed that OA treatments maintained related to TSS may be attributed to different
flesh firmness, delayed softening and extended species and storage condition.
Table 2. Firmness (N) of ʻAprikozʼ apricots treated with OA during MAP storage at 0°C
Treatment 0d 10 d 20 d 30 d 40 d Means
Control 36.10 31.46 29.64 29.20 27.13 30.71 c*
1 mM 36.10 35.87 34.23 33.89 32.04 34.43 a
2 mM 36.10 35.00 33.65 32.55 30.31 33.52 ab
4 mM 36.10 33.24 32.30 29.55 28.96 32.03 bc
8 mM 36.10 31.96 31.71 29.34 29.30 31.68 c
Means 36.10 a 33.51 b 32.30 bc 30.91 cd 29.55 d
*Means followed by the same letter in the same row and column are not statistically significant (P<0.05); d: days.
Table 3. TSS and TA of ʻAprikozʼ apricots treated with OA during MAP storage at 0°C
Control 11.53 11.63 11.00 10.80 9.97 10.99 bc*
1 mM 11.53 11.70 11.47 11.20 10.50 11.28 a
2 mM 11.53 10.93 10.87 10.63 10.47 10.89 bc
TSS
4 mM 11.53 11.00 10.97 10.70 9.57 10.79 c

8 mM 11.53 11.90 11.43 10.57 10.27 11.14 ab
Means 11.53a 11.39ab 11.19b 10.78c 10.15d
Control 1.10 1.02 1.00 0.89 0.87 0.98 ab*
1 mM 1.10 1.07 1.02 0.93 0.92 1.01 a
2 mM 1.10 1.04 0.96 0.92 0.86 0.98 ab
TA
4 mM 1.10 0.95 0.94 0.90 0.81 0.94 b

8 mM 1.10 0.94 0.91 0.90 0.77 0.92 b
Means 1.10 a 1.00 a 0.97 a 0.91 b 0.85 b
149
Ethylene production and respiration rate. In are given in Table 5. The effects of storage
the present study, the effect of the treatments period and treatments on L*, a* and b* value of
on ethylene production (μL/kg h) and fruit skin were statistically significant,
respiration rate (mL CO2/kg h) was significant (P<0.05). L* value, which shows fruit
(P<0.05). The highest ethylene production brightness of fruit skin, decreased during the
(0.58 μL/kg h) was determined from untreated storage period. The highest L* value was 72.93
fruit, whereas the lowest ethylene production with OA 8 mM treatment. The a* and b* values
(0.46 μL/kg h) was detected in 8 mM OA- of the fruit generally showed an increase during
treated fruit. The OA treatments influenced the storage. All treatments showed similar
ethylene production of apricots depending on characteristics of the a* value when compared
dose levels. The average ethylene production of to the control group. The highest a* (-6.80) and
fruit decreased with increasing doses of OA lowest b* (44.68) values were obtained from 4
showing its obvious effect on ethylene mM OA treated fruit. The skin color of apricots
biosynthesis (Table 4). Respiration rate is an is one of the symbols of fruit senescence. In
important factor in maintaining quality during general, the color of the skin changes from
cold storage and shelf life of fruit. For this yellow-green (at harvest) to orange-yellow (at
reason, it is important to reduce the respiration the end of storage). OA-treated fruit turned to
rate of the apricots during storage. It can be orange-yellow slowly compared to control fruit
seen from data that the respiration rates of OA during storage. Turning color to dark-orange
treated fruit are suppressed better than control has been associated with ripening of apricot.
group. The lowest respiration rate (44.64 mL OA delayed the ripening process of climacteric
CO2/kg h) was detected in 4 mM OA-treated fruit such as mango (Zheng et al., 2007a) and
fruit. Similarly in a previous study, the peach (Zheng et al., 2007b), due to the
decreased ethylene production by OA inhibition of ethylene production.
contributed to the delaying of ripening of plum Sensory analyses. Storage period and treat-
fruit, and reported that reduced ethylene ments affected significantly the external
production in OA-treated fruit might be appearance, taste scores and internal browning
ascribed to the reduced 1-aminocyclopropane- of apricots during storage (p<0.05).
1-carboxylic acid synthase (ACS) activity (Wu OA treated apricots preserved their external
et al., 2011). In addition, it was reported that appearance and taste values better than control
OA treatments reduced respiration rate of fruit (Table 6).
peach fruit, and inhibited the ethylene The average highest external appearance (8.79)
production rates in mango and plum (Huang et and taste (4.83) scores were obtained from 1
al., 2013). mM OA treatment during storage.
Fruit skin color. The results of the effects of
OA treatments on the change of fruit skin color
Table 4. Ethylene production (μL/kg h) and respiration rate (mL CO2/kg h) of ʻAprikozʼ apricots treated with OA
during MAP storage at 0°C
Control 0.54 0.54 0.63 0.73 0.48 0.58 a*
Production
1 mM 0.36 0.56 0.52 0.67 0.57 0.54 ab

Ethylene
2 mM 0.37 0.42 0.42 0.80 0.65 0.53 ab

4 mM 0.40 0.50 0.40 0.50 0.67 0.50 ab
8 mM 0.43 0.58 0.42 0.45 0.45 0.46 b
Means 0.42 c 0.52 abc 0.48 bc 0.63 a 0.56 ab
Control 95.97 67.46 48.90 32.77 34.74 55.97 a*
Respiration
1 mM 75.52 42.99 48.01 41.14 34.79 48.49 bc

2 mM 86.28 59.50 50.39 36.55 33.80 53.31 ab
Rate
4 mM 80.01 35.38 33.33 43.25 31.25 44.64 c

8 mM 84.71 52.83 35.70 39.77 34.35 49.47 abc
Means 51.63 a 43.27 b 38.70 c 33.79 cd 34.50 d
150
Table 5. Change color (L*, a*, b*) of ʻAprikozʼ apricots treated with OA during MAP storage at 0°C
Control 74.07 72.32 73.65 71.90 70.81 72.55 ab*
1 mM 74.07 73.47 72.75 71.54 70.51 72.47ab
2 mM 74.07 73.14 73.35 72.26 68.24 72.21 b
L*
4 mM 74.07 72.65 73.30 71.84 68.13 71.99 b

8 mM 74.07 74.28 74.22 73.43 68.65 72.93a
Means 74.07 a 73.17 c 73.45 b 72.19 c 69.26 d
Control -11.79 -9.75 -4.32 -6.02 -4.85 -7.34ab*
1 mM -11.79 -8.37 -4.56 -5.60 -7.69 -7.60b
2 mM -11.79 -7.53 -5.10 -4.95 -6.51 -7.17ab
a*
4 mM -11.79 -7.11 -5.04 -4.54 -5.53 -6.80a

8 mM -11.79 -6.87 -5.39 -7.10 -6.41 -7.51ab
Means -11.78 d -7.92 c -4.88 a -5.63 ab -6.19 b
Control 43.99 45.18 47.64 46.84 45.16 45.76a*
1 mM 43.99 47.47 45.71 46.89 44.46 45.70a
2 mM 43.99 45.20 46.96 45.53 43.00 44.93b
b*
4 mM 43.99 43.85 45.38 46.94 43.27 44.68b

8 mM 43.99 47.01 47.34 46.19 43.89 45.68a
Means 43.99 c 45.74 b 46.60 a 46.47 a 43.95 c
Table 6. The external appearance, taste and internal browning scores of fruit during MAP storage at 0°C
Control 9.00 9.00 9.00 7.45 8.33 8.12 d*
Appearance
1 mM 9.00 9.00 9.00 8.60 8.05 8.79 a

External
2 mM 9.00 9.00 9.00 8.50 7.96 8.71 b

4 mM 9.00 9.00 9.00 8.20 7.80 8.63 c
8 mM 9.00 9.00 9.00 8.14 6.15 8.59 c
Means 9.00 a 9.00 a 9.00 a 8.18 b 7.66 c
Control 5.00 5.00 5.00 4.20 3.20 4.42 c*
1 mM 5.00 5.00 5.00 4.86 4.30 4.83 a
Taste
2 mM 5.00 5.00 5.00 4.25 4.00 4.65 b

4 mM 5.00 5.00 5.00 4.10 4.00 4.62 b
8 mM 5.00 5.00 4.70 4.05 3.80 4.57 b
Means 5.00 a 5.00 a 4.94 a 4.29 b 3.86 c
Control 0.00 0.00 0.60 1.50 1.90 0.84 a*
1 mM 0.00 0.00 0.00 1.10 1.40 0.50 c
browning
Internal
2 mM 0.00 0.00 0.00 1.20 1.60 0.56 bc

4 mM 0.00 0.00 0.00 1.40 1.80 0.64 bc
8 mM 0.00 0.00 0.00 1.60 1.70 0.66 c
Means 0.00 c 0.00 c 0.12 c 1.36 b 1.68 a
External appearance: 1-3: unmarketable, 5: marketable, 7: good, 9: very good; Taste: 1: very bad, 2: bad, 3: medium, 4:
good, 5: very good; Internal browning: 0: healthy, 1: 1-10%, 2: 11-33%, 3: 33-66%, 4: 66-100%.
Control fruit gave the lowest external limiting factor for marketable quality of
appearance (8.12) and taste (4.42) scores. apricots (Koyuncu et al., 2010).
Internal browning of fruit increased compared Gas composition. The CO2 concentration in the
to the initial values at the end of cold storage packages increased, while a decreasing was
regardless of treatment, but the lowest value found in O2 concentration compared to initial
was determined in 1 mM OA treatment. The levels during storage period (Table 7). The
OA treatments limited internal browning effects of storage period and treatments on O2
incidence of apricots (Table 6). It can be said and CO2 values in MAP were significant
that internal browning is the important and (P<0.05). The highest average O2 (15.06%) and
151
the lowest CO2 (5.65%) concentrations were characteristics and sensory quality properties of
measured in packages of 1 mM OA treatment. apricot fruit during cold storage and shelf-life. Polish
Journal of Food and Nutrition Sciences, 67 (2): 159-
This means that the 1 mM dose of OA 166.
suppressed respiration rate of apricots better Huang H., Jing G., Guo L., Zhang D., Yang B., Duan,
than the other treatments. Generally, OA X., Ashraf M., Jiang Y., 2013. Effect of oxalic acid on
treatments decreased respiration rate of apricots ripening attributes of banana fruit during storage.
compared to control group according to gas Postharvest Biology and Technology, 84: 22-27.
Jing Y., Ma X., Jin P., Zhu X., 2018. Effects of harvest
compositions in MAP. Our findings related to maturity on chilling injury and storage quality of
respiration rate (Table 4), which indicates apricots. Journal of Food Quality, 1-7 (4954931).
suppressing effect of OA on respiration, Koyuncu M.A., Dilmaçünal T., Özdemir Ö., 2010.
support present results. Modified and controlled atmosphere storage of
apricots. Acta Horticulturae, 876: 55-66.
Table 7. The O2 (%) and CO2 (%) composition of MAP Li P., Zheng X., Liu Y., Zhu Y., 2014. Pre-storage
during storage at 0°C application of oxalic acid alleviates chilling injury in
mango fruit by modulating proline metabolism and
Treatment 10 d 20 d 30 d 40 d Means energy status under chilling stress. Food Chemistry,
Control 15.55 14.55 14.15 11.60 13.96ab* 142:72-78.
1 mM 15.85 15.90 15.25 13.23 15.06 a Martinez-Espla A., Zapata P.J., Valero D., Garcia-
2 mM 15.95 14.35 14.11 12.96 14.34 ab Viguera C., Castillo S., Serrano M., 2014. Pre-harvest
O2
4 mM 15.80 15.45 13.90 13.63 14.70 ab application of oxalic acid increased fruit size,
bioactive compounds, and antioxidant capacity in
8 mM 15.85 13.15 12.65 10.90 13.14 b
sweet cherry cultivars (Prunus avium L.). Journal of
Means 15.80a 14.68ab 14.01bc 12.46c
Agricultural and Food Chemistry, 62: 3432-3437.
Control 5.88 6.30 7.10 8.65 6.98 a* Muradoğlu F., Pehlivan M., Gündoğdu M., Tuncay K.,
1 mM 4.15 4.90 6.30 7.25 5.65 b 2011. Iğdır yöresinde yetiştirilen bazı kayısı (Prunus
2 mM 5.70 5.95 7.40 8.20 6.81 a armeniaca L.) genotiplerin fizikokimyasal özellikleri
CO2
4 mM 5.85 6.15 7.30 7.90 6.80 a ile mineral içerikleri, Iğdır Üniversitesi Fen Bilimleri
8 mM 4.82 6.15 7.30 8.35 6.66 a Enstitüsü Dergisi, 1 (1): 17-22.
Özdoğru B., Şen F., Acarsoy Bilgin N., Mısırlı A., 2015.
Means 5.28 c 5.89 c 7.08 b 8.07a
Bazı sofralık kayısı çeşitlerinin depolanma sürecinde
*Means followed by the same letter in the same row and
fiziksel ve biyokimyasal değişimlerinin belirlenmesi.
column are not statistically significant (P<0.05); d: days.
Ege Üniviversitesi Ziraat Fakültesi Dergisi, 52 (1):
23-30.
CONCLUSIONS Razavi F., Hajilou J., 2016. Enhancement of pos-tharvest
nutritional quality and antioxsidant capacity of peach
In conclusion, all doses of OA gave better fruits by pre-harvest oxalic acid treatment. Scientia
Horticulturae, 200: 95-101.
results than control group in terms of some
Razzaq K., Khan AS., Malik AU., Shahid, M., 2015.
quality parameters. Especially, 1 mM dose of Effect of oxalic acid application on Samar Bahisht
OA was the most effective treatment for Chaunsa mango during ripening and pos-tharvest.
decreasing weight loss and maintaining fruit LWT-Food Science and Technology, 63: 152-160.
flesh firmness, TA and sensory quality. The Sayyari M., Valore D., Babalar M., Kalantari S., Zaptara
P.J., Serrano M., 2010. Pre-storage oxalic acid
results suggest that OA has the potential to
treatment maintained visual quality bioactive
extend the storage life of apricot by delaying compounds, and antioxidant potential of pomegranate
quality loss. Based on our results, it can be after long-term-storage at 2°C. Journal of Agricultural
indicated that OA could maintain fruit firmness and Food Chemistry, 58: 6804-6808.
and delayed quality loss of apricot by suppre- Wu F., Zhang D., Zhang H., Jiang G., Su X., Qu H., Jian
ssion of ethylene production and respiration Y., Duan X., 2011. Physiological and biochemical
response of harvested plum fruit to oxalic acid during
rate. ripening or shelf-life. Food Research International, 44:
1299-1305.
REFERENCES Zheng X., Tian S., Gidley M.J., Yue H., Li B., 2007a.
Effects of exogenous oxalic acid on ripening and
Abd El Wahab S.M., 2015. Apricot pos-tharvest fruit decay incidence in mango fruit during storage at room
quality, storability and marketing in response to pre temperature. Post-harvest Biology and Technology,
harvest application. Middle East Journal of 45: 281-284
Agriculture, 4 (2): 347-358. Zheng X., Tian S., Meng X., Li B., 2007b. Physiological
Ezzat A., Ammar A., Szabó Z., Nyéki J., Holb I.J., 2017. and biochemical responses in peach fruit to oxalic
Post-harvest treatments with methyl jasmonate and acid treatment during storage at room temperature.
salicylic acid for maintaining physico-chemical Food Chemistry, 104: 156-162.
152
EFFECTS OF PRE-HARVEST RETAIN TREATMENTS WITH MAP ON

COLD STORAGE QUALITY OF SWEET CHERRY CV. ʻ0900 ZIRAATʼ
Atakan GUNEYLI1, Mehmet Ali KOYUNCU2, Cemile Ebru ONURSAL3, Derya ERBAS2,
Melike CETİNBAS1, Sinan BUTAR1, Fatma KOYUNCU2
1
Fruit Research Institute, Isparta, Turkey
2
Suleyman Demirel University, Faculty of Agriculture, Department of Horticulture, Isparta, Turkey
3
Batı Akdeniz Agrıcultural Research Institute, Antalya, Turkey
Abstract
Effect of pre-harvest treatments of ReTain [active ingredient: Aminoethoxyvinlglycine (AVG) 15 %] on cold storage
quality of sweet cherry cv. ʻ0900 Ziraatʼ grafted on Gisela 5 (Prunus cerasus × Prunus canescens) rootstock was
investigated. For this purpose ReTain within concentrations of 50, 100, and 150 mg/L was applied as a spray 25 days
before harvest. Fruits were harvested at the optimum harvest date and stored in modified atmosphere packages (MAP)
at 0°C temperature and 90±5% relative humidity conditions for 6 weeks. Weight loss, fruit skin colour, fruit firmness,
total soluble solid and titratable acidity content were determined during the storage period. In addition, sensory
analysis was performed. Weight loss increased in all treatments during storage. Titratable acidity of fruit decreased
with increasing storage period, but the highest average acidity was determined in fruit treated with 100 mg/L ReTain.
The treatment of 50 mg/L of ReTain maintained fruit firmness better than those of others. These results indicated that
pre-harvest ReTain treatment can be a tool for maintaining some quality attributes of sweet cherry during cold storage.
Key words: ʻ0900 Ziraatʼ, sweet cherry, MAP, ReTain, cold storage.
INTRODUCTION needed, which have to be considered as healthy

and environmentally friendly (Serrano et al.,
Sweet cherry is one of the most important crops 2005). Among these technologies, the use of
of Turkey, constituting approximately 20-25% modified atmosphere packaging (MAP) has
of world production with the production been reported to be effective in cherry storage
capacity of 599,650 tons (2016 year) (FAO, (Spotts et al., 2002; Tian et al., 2004;
2018). In addition to production, sweet cherry, Kupferman et al., 2005). The MAP is used for
cv. ʻ0900 Ziraatʼ, is an important species for keeping the postharvest quality of fruit by
export of Turkey (Eştürk et al., 2012). But fruit lowering the respiration rate of fruit
losses after harvest are still high depending on (decreasing O2 and increasing CO2) (Kader et
the species, harvest methods, transporting and al., 1989). MAP extends the postharvest life of
marketing conditions and length of storage. cherry and decreases decay resulting from
Sweet cherry decays rapidly after harvest, so softening, and maintain green stem colour
the consumption period is short and limited. (Kupferman et al., 2005). As in other fruit,
The main causes of fruit deterioration are water plant growth regulators are used to increase
loss, stem browning, softening, colour changes quality and to extend post-harvest life of sweet
and pitting (Bernalte et al., 2003). Water loss cherries (Onursal et al., 2013). ReTain is, a
and firmness are important quality attributes of commercial product that includes 15% AVG, a
cherries, and are directly related to storability plant growth regulator, and effects harvesting
(Martinez-Romero et al., 2006). Increase in criteria (Clayton et al., 2000).
these postharvest losses can cause great Aminoethoxyvinylglycine (AVG) is a natural
economic losses (Esti et al., 2002). compound produced in plant tissues (Rath et
Many pre- and post-harvest technologies have al., 2006). Researchers have shown that pre-
been used to delay these losses, but the use of harvest AVG treatment has maintained skin
chemicals has been limited in some countries. colour and firmness of some fruits (Jobling et
So, alternative technologies for preservation are al., 2003; McGlasson et al., 2005; Çetinbaş et
153
al., 2012). This research was carried out to 96801, Hanna, UK) and expressed as a per-
determine the effects of pre-harvest ReTain centage (%). Titratable acidity (TA) was
(AVG) treatment on quality of ʻ0900 Ziraatʼ determined by an automatic titrator (Mettler
sweet cherry during cold storage. Toledo T50, Switzerland) and expressed as
grams of malic acid equivalent per 100 g fresh
MATERIALS AND METHODS weight.
Overall acceptability was rated on a hedonic
Plant material and retain treatment scale of 0-2 (0: good commercial quality, 1:
The study was conducted at Fruit Research some damage but still commercially salable, 2:
Institute, Isparta-Turkey. The uniform trees, cv. not commercially salable), described by Feng et
ʻ0900 Ziraatʼ sweet cherry, on Gisela-5 al. (2004). Taste was rated on a scale of 1-5 (1:
(Prunus cerasus × Prunus canescens) very bad, 2: bad, 3: medium, 4: good, 5: very
rootstock, were used. Standard cultural good), described by Erbaş and Koyuncu
practices including, thinning and pesticide (2016).
sprays were provided to the trees. The experiment was set up according to the
For pre-harvest treatment, 50, 100 and 150 factorial randomized design with 3 replications
mg/L Retain and Tween-20 (0.01%) (as a (5 kg fruit per replication). Data were subjected
surfactant) were sprayed on sweet cherry trees to analysis of variance (ANOVA, JMP7),
as well as distilled water with Tween-20 (pre- means were separated by means of LSD test
harvest control group) 25 days before (P<0.05, 0.01, 0.001).
commercial harvest (when fruits turned to
straw-color). Cherries were harvested at RESULTS AND DISCUSSIONS
commercial harvest maturity from an orchard
and transported to the laboratory immediately. Weight loss which is the most important factor
Harvested cherries were dipped in cool water limiting the storage period, increased during
(0-2°C) for 8-10 minutes for pre-cooling. After storage in all treatments, especially in control
pre-cooling, cherries were placed on the MAP treatment (Table 1). In this study, the weight
(Xtend) and stored at 0°C and 90±5% relative loss of cherries was low levels because of the
humidity for 6 weeks. water vapour permeability properties of the
Chemical and physical analysis MAP. The effects of storage periods on weight
Weight loss of cherries was measured over 5 kg loss were statistically significant but there was
fruits in each replicate and expressed as the no significant effect of treatments (P<0.01)
percentage of loss of weight with respect to the (Table 4). At the end of storage, the weight loss
initial weight. Weight loss was determined by of cherries was between 2.36% (control) and
the formula; [(First weight - Last weight) / First 2.07% (100 mg/L) (Table 1). The results found
weight] × 100. in this research agree with those of previous
Fruit skin color was determined using a studies (Üstünel et al., 2008; Machado et al.,
colorimeter (CR400, Minolta Co., Japan) over 2010).
15 fruits in each replicate. Minolta color
measurement apparatus was calibrated Table 1. Effect of pre-harvest ReTain treatment on
according to the standard white calibration weight loss of ʻ0900 Ziraatʼ cherries stored at MAP
plate (Y: 92.3, x: 0.3136 and y: 0.3194). The 1w 2w 3w 4w 5w 6w Means
values were expressed by the CIEL* Control 1.69 2.04 2.05 2.19 2.78 3.38 2.36ns
(brightness-darkness), a* (+ a*: red, ** - a*: 50 mg/L 1.10 1.66 1.87 2.64 2.92 2.97 2.19
green) and b* (+ b*: yellow, - b*: blue). 100 mg/L 1.57 1.61 2.22 2.27 2.34 2.42 2.07
Fruit firmness was determined using a texture 150 mg/L 1.64 2.01 2.03 2.21 2.81 3.18 2.31
Means 1.50C* 1.83C 2.04BC 2.33ABC 2.71AB 2.99 A
analyzer (Guss FTA Type GS14, Strand, South *Means followed by different letters with in the same
Africa). It was defined as the maximum load row is significantly different at P<0.01; ns:
required to penetrate the probe (5 mm nonsignificant; w: weeks.
diameter) into the fruit flesh (6 mm). The
results were expressed in Newton (N). Softening of cherries reduces both market value
Total soluble solids (TSS) content was and consumer acceptability. Changes in fruit
measured using a digital refractometer (HI firmness during cold storage are presented in
154
Table 2. The effect of storage time and (0.68%) in 100 mg/L dose of ReTain. 150
treatments on the fruit firmness were mg/L dose of ReTain treatment was the best
statistically significant (P<0.01) (Table 4). treatment for maintaining the TA. During
Fruit firmness decreased with increasing storage, TA values were continuously
storage periods but generally, fruit softening decreased (Table 3). Our results are in
were minimized by ReTain treatment agreement with Khorshidi et al. (2011) who
regardless of doses. At the end of the storage, reported that sweet cherries, a nonclimacteric
100 mg/L doses of ReTain was the best fruit, use sugars and acids for respiration, so
treatment for maintaining of firmness compared TA can decrease depending on storage period.
to other treatments. The positive effects of The colour of sweet cherries is probably the
ReTain treatment combined with the MAP on main quality attribute considered by
fruit firmness were recorded in this study. In consumers. Changes in fruit skin color during
previous studies, pre- and post-harvest ReTain cold storage are given in Figure 1. L* values,
treatments maintained fruit firmness better than which shows fruit brightness-darkness,
control groups during the storage (Drake et. al., decreased throughout storage. Fruit in control
2006; Kharoshaki et. al., 2008; Lara, 2013). group lost their brightness more than ReTain
These results can be explained by delaying treatments. While the highest mean L* value
water loss and maintaining pectin level in the was obtained from the dose of 100 mg/L
cell wall of fruit related to ReTain. (31.55), the greatest decrease (27.09) occurred
A large part of the TSS is composed of sugars, in control group at the end of the storage.
and change of TSS during the storage is due to Similar trend was also observed for a* and b*
changes in fruit carbohydrate structure. The values of skin colour. The a* (+a*: redness, -a*:
water loss of fruit affects fruit TSS content, and greenness) and b* (+b*: yellowness, -b*:
generally it increase (Kader, 1989). But in this blueness) values steadily decreased during the
study, the weight loss of cherries was very low storage. However, the amount of decrease in
(Table 1) so the increasing of TSS was low, ReTain-treated cherries less than control
too. The TSS of fruit increased at the end of 42 treatment (Figure 1). ReTain treatments have
days compared to initial values, with been reported to be beneficial in maintaining
fluctuation during storage (Table 3). No fruit colour (Onursal et al., 2013).
significant interaction existed between The results of the sensory analyses are
treatments and storage periods, but differences presented in Figure 2. Overall acceptability and
between storage periods and treatments for TSS taste decreased during storage. Storage period
values were significant (Table 4). The lowest and treatments affected significantly the
change in TSS value according to initial values external appearance and taste scores of cherries
was obtained from dose of 150 mg/L (Table 3). during storage (p<0.01) (Table 4). ReTain-
Similar observations were recorded by Remon treated cherries (especially 100 mg/L dose)
et al. (2000) and Onursal et al. (2013) for preserved their taste scores better than control
cherries. The main factor of taste formation in fruit. The better taste was obtained from 100
fruit and vegetables are TA (Karaçalı, 2009). mg/L dose during 5 weeks, while control fruit
Storage periods and treatments significantly had a bad taste after 3 weeks of storage. Similar
affected TA contents of cherries (Table 4). results were reported by Drake et al. (2006) and
During the 6-weeks cold storage, the lowest Olmstead et al. (2012).
average (0.65%) TA value was obtained from
50 mg/L dose of ReTain, and the highest
Table 2. Effect of pre-harvest ReTain treatment on fruit firmness of ʻ0900 Ziraatʼ cherries stored at MAP
0w 1w 2w 3w 4w 5w 6w Means
Control 2.65 2.55 2.47 2.45 2.39 2.17 1.68 2.34b*
50 mg/L 2.65 2.60 2.59 2.57 2.49 2.48 2.52 2.56a
100 mg/L 2.70 2.34 2.71 2.71 2.75 2.67 2.37 2.61a
150 mg/L 2.52 2.43 2.39 2.40 2.35 2.30 1.99 2.34b
Means 2.64A* 2.48AB 2.54A 2.53A 2.50AB 2.41BC 2.14C
*Means followed by different letters with in the same row and column are significantly different at P<0.01; w: weeks.
155
Table 3. Effect of pre-harvest ReTain treatment on total soluble solid content and titratable acidity
of ʻ0900 Ziraatʼ cherries stored at MAP
0w 1w 2w 3w 4w 5w 6w Means
Control 15.81 16.07 16.37 17.00 16.50 16.07 15.93 16.25 b*
50 mg/L 15.43 15.80 16.67 16.77 17.97 16.98 15.70 16.34 a
TSS 100 mg/L 16.83 17.40 17.83 17.87 18.07 17.97 17.43 17.63 a
150 mg/L 15.60 16.37 16.80 17.43 17.23 16.63 15.63 16.53 b
Means 15.92AB* 16.41BC 16.92B 16.25C 17.27BC 16.91A 16.17BC
Control 0.79 0.72 0.68 0.63 0.63 0.60 0.57 0.66 bc
50 mg/L 0.76 0.72 0.65 0.64 0.60 0.59 0.58 0.65 c
TA 100 mg/L 0.80 0.75 0.69 0.68 0.65 0.61 0.60 0.68 a
150 mg/L 0.78 0.74 0.67 0.65 0.64 0.61 0.61 0.67 ab
Means 0.78A* 0.73B 0.67C 0.65C 0.63D 0.61D 0.59D
*Means followed by different letters with in the same row and column are significantly different at P<0.01; w: weeks.
Table 4. Anova for dependent variables for treatments, storage period and their interactions for cherries
Weight loss Firmness Overall acceptability Taste L* a* b* TSS TA
SP ** ** ** ** ** ** ** ** **
T ns ** ** ** ** ** ** ** **
SP × T ns ns ns ns ** ** ** ns ns
SP: Storage period; T: Treatments; ns: represents non-significance at P< 0.05; **Represents significance at the 0.01;
TA: Titratable acidity; TSS: Total soluble solid.
Control 50mg/L 100mg/L 150mg/L

Control 50mg/L 100mg/L 150mg/L
35 2,5
34
Overall Acceptability (0-2)
33 2,0
32
31
1,5
L*
30
29
1,0
28
27
26 0,5
LSDsp: 0.272; LSD t: 0.206; LSD t×sp: 0.544
25
0 1 2 3 4 5 6 0,0
0 1 2 3 4 5 6
29
6
27
5
25
4
23
Taste (1-5)
a*
21 3
19 2
17 1
LSDsp: 0.335; LSD t: 0.253; LSD t×sp: 0.669
15
0 1 2 3 4 5 6 0
0 1 2 3 4 5 6
16 Storage periods (weeks)
15
14 Figure 2. Effect of pre-harvest ReTain treatment on
13 sensory analysis of ʻ0900 Ziraatʼ cherries stored at MAP
12 Vertical bars represent the standard error of the mean
(n=3). Overall acceptability: 0: good commercial quality,
b*
11
10 1: some damage but still commercially salable, 2: not
9
commercially salable; Taste scores: 1: very poor, 2: poor,
8 LSDsp: 0.494; LSD t: 0.373; LSD t×sp: 0.987
7
3: mild, 4: good, 5: excellent
0 1 2 3 4 5 6
Storage periods (weeks) CONCLUSIONS
Figure 1. Effect of pre-harvest ReTain treatment on fruit
skin color (L*, a* and b*) of ʻ0900 Ziraatʼ cherries stored At the end of the 6 weeks of storage, AVG
at MAP (SP: Storage period; T: Treatments) treatment was more effective for maintaining
156
postharvest quality of cherries compared to vegetables in Critical Reviewes in Food Science and
control treatment. Treated cherries with AVG Nutrition, 28 (1): 1-30.
Karaçalı İ., 2009. Bahçe Urunlerinin Muhafaza ve
delayed colour changes, softening and loss of Pazarlanması. Ege University, Faculty of Agriculture,
acidity. In addition, AVG treatment was Publication No: 494, İzmir-Türkiye, 486.
effective in the maintaining sensory quality Kharoshaki M.Z.K., Ashari M.Z., Fattahi M., 2008.
during the storage. Especially 100 mg/L AVG Effect of exogenus putracine on post-harvest life of
as pre-harvest treatment was the most effective sweet cherry (Prunus avium) fruit, cultivar ʻSurati-e
Hamedaʼ. Journal of Applied Horticulture, 10 (2):
dose and it can be used to maintain sweet 154-157.
cherry postharvest quality during storage of 35 Khorshidi S., Davarynejad G., Tehranifar A., Fallahi E.,
days at 0°C. 2011. Effect of modified atmosphere packaging on
chemical composition, antioxidant activity,
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158
AN INVESTIGATION ON EFFECTS OF DRY AND WET CLIMATE

CONDITIONS ON PISTACHIO (PISTACIA VERA) YIELD IN MIDDLE
EUPHRATES BASIN, SOUTHEAST OF TURKEY
Mehmet Ali ÇELİK1, Muhittin KULAK2, 3, Hakan CETİNKAYA4, Mehmet KOC4, Ali
GÖCERİ5, Salman ÖZÜPEKÇE1
1
Kilis 7 Aralık University, Faculty of Arts and Sciences, Department of Geography, Kilis, Turkey
2
Kilis 7 Aralık University, Faculty of Arts and Sciences, Department of Biology, Kilis, Turkey
3
Kilis 7 Aralık University, Yusuf Serefoglu School of Health, Department of Nursing, Kilis, Turkey
4
Kilis 7 Aralık University, Faculty of Arts and Sciences, Department of Horticulture, Kilis, Turkey
5
Kahramanmaras Sutcu Imam University, Graduate School of Natural and Applied Sciences,
Kahramanmaras, Turkey
Abstract
Extreme conditions in the climate play an important role on the plant growth and yield of crops. Meteorological
drought is one of these extreme climate conditions. In this study, 16 days photosynthetic activity of pistachios was
investigated by integrating both field research and modern techniques (remote sensing and statistical methods).
Furthermore, the effects of climatic factors on the yield of pistachio were analysed. Along with the present study, the
relationships between pistachio and dry and wet climatic conditions were presented. According to the results of the
drought analysis of both Birecik (a district in the south-eastern parts of Turkey) and Gaziantep (a province in the south-
eastern parts of Turkey) stations, pistachio is significantly affected by extreme climatic conditions. The photosynthetic
activity of pistachio increased during the periods of wet climatic conditions.
Key words: climate, drought, photosynthetic activity, Pistacia vera.
INTRODUCTION significant effects on agricultural production

(Box et al., 1989; Alexandrov and
Turkey's southern line constitutes one of the Hoogenboomb, 2000; Chmielewski and Rötzer,
sensitive areas where short or long-term 2001; Ichii et al., 2002; IPCC, 2007; Sanchísa
climate change in the world can be and Feijoo Bello, 2009; Cheng and Wu, 2011).
experienced. Southeast Anatolia is particularly The study of the impact of climate change on
vulnerable to climate change (Cosun and the phenology of agricultural products has a
Karabulut, 2009). One of the most important major precaution in terms of food supply and
consequences of climate change is the increase socio-economic in the world with a population
in extreme climatic conditions. The arid period, of over 7 billion. One of the most worried
in which the humid period with extreme issues for the future is the danger of
precipitation and the amount of rainfall and agricultural production due to extreme climatic
humidity are well below the average, is the conditions and the resulting scarcity. The
extreme climate. Depending on the climate answer to the question of how global climate
change, there is an increase in the frequency of change will impact on agricultural production
extreme climatic conditions in the area where is among the most important concerns of
Gaziantep, Şanlıurfa and Adıyaman province concern all over the world (Tubiello and
are located. Fischer, 2007).
Depending on this increase, significant changes Pistacia vera (pistachio) belonging to
occur in the phenology and yield of agricultural Anacardiaceae is of the characterized crop for
products. Also, many previous studies report the South-eastern parts of Turkey. Southeastern
that changes in climate varieties can lead to Anatolia Region of Turkey is the most
phenological changes in agricultural products pistachio producing region. This region is
and changes in phenological cycles will have followed by the Mediterranean and Aegean
159
regions (Külekçi and Aksoy, 2011; Ertürk et of 1 km are scanning the globe twice a day. The
al., 2015). More than 80% of the pistachio- first of these takes place at around 10 pm local
planted areas in the Southeast are in Gaziantep time and the other at 2.30 pm. All these
and Şanlıurfa. Gaziantep and Sanliurfa also collected data are collected via various earth
covers a portion of 78% of the pistachio field in stations (Karabulut, 2006).
Turkey. Therefore, any possible incident
regarding drought that may emerge in Sanliurfa
and Gaziantep affect significantly the
production of pistachios in Turkey. In Turkey,
pistachio plantings of that period it was first
built in 1960 production was 34 tons (Gul and
Akpinar, 2006) in a humid climate today late in
the period, this figure rises to 150 tons. In the
years when the climate is arid (for example in
2014), pistachio production has been
determined as 80 tons. Therefore, it is
necessary to extensively analyze the effect of
climate conditions on the production of
pistachio nuts. The results of the drought
analysis applied to the climate data obtained
Figure 1. Location map of study area
from the General Directorate of Meteorology
(MGM) show that both in Sanliurfa and in
MODIS NDVI images with a spatial resolution
Gaziantep, the years of 2012 and 2013 are
of 250 m are published in 16 day composites
humid. On the other hand, in 2014, it is
for drawing twice a day. Radiometric resolution
observed that dryness occurs in these two
of the images taken twice a day for 16 days is
stations where pistachio production is highest.
published by compositing the images that are
Significant changes in Turkey's pistachio yield
free from brdf and cloud. The MODIS NDVI
during dry and wet climate periods are great
images, consisting of 4800 rows and 4800
concern.
columns, are capable of analyzing the variation
In this context, the current study was designed
in vegetation activity over a wide area (Çelik
to investigate the possible influence of dry and
and Sonmez, 2013).
wet climate conditions on the pistachio nut
Another plant index model that has recently
yield from two stations, namely Gaziantep and
been used frequently is Enhanced Vegetation
Sanliurfa, which are considered as main
Index (EVI). EVI images are not affected much
pistachio suppliers for Turkey.
by aerosol and cloudiness. The EVI images
provided by MODIS to its users are calculated
by the following formula (Huete et al., 2002;
Galvão et al., 2011; Zhang et al., 2014):
For the current study, three provinces in the
south-eastern parts of Turkey (Gaziantep,
EVI = G × (Near Infrared-Red) / (Near Infrared
Sanlıurfa and Adıyaman) were selected to due
+ C1 × Red-C2 × Blue + L)
to their potential pistachio production (Figure
1). In this study, during the dry and wet
The gain factor used in the formula is 2.5, L =
climatic periods, the yield and phenology of
1, C1 = 6, C2 = 7.5. With this formula values
pistachios were monitored. In this context,
appear in the band interval ranging from -1 to
Moderate Resolution Imaging
+1. EVI displays, which are not much affected
Spectroradiometer (MODIS) and National
by atmospheric conditions, do not show too
Oceanic and Atmospheric Administration
much deviation during the year as a trend.
(NOAA) Advanced Very High Resolution
In order to determine the periods when the
Radiometer (AVHRR) satellite data were used.
climate was arid and humid, Standardized
The meteorological satellites carrying the
Rainfall Index (SPI) developed by Mckee et al.
NOAA-AVHRR sensor with spatial resolution
(1993) was used and SPI was calculated by the
160
following formula; low yield of pistachio could be consequences of
X i X iort periodicity, inadequate pollination, fertilization,
SPI
water stressful conditions and traditional
According to the results of the formula, when cultural practices.
the index falls below zero, it is considered as
Table 1. Area harvested, yield and production of
the beginning of the month, and when the index
pistachio between 2000-20016 in Turkey
is positive, the month is considered as the end
of the drought. Years Area harvested Yield Production
(ha) (t/ha) (t)
RESULTS AND DISCUSSIONS 2000 36349 2.0633 75000
2001 36999 0.8108 30000
Monitoring of agricultural areas in short 2002 37428 0.9351 35000
periods is of great importance in terms of 2003 37570 2.3955 90000
revealing the effects of environmental 2004 37572 0.7985 30000
phenomena (such as climatic and biotic 2005 40000 1.5000 60000
factors). In this regard, remote sensing 2006 40000 2.7500 110000
technology has many advantages over 2007 40661 1.8056 73416
conventional methods since traditional methods 2008 40954 2.9329 120113
are inadequate in detecting short-term changes 2009 43063 1.8994 81795
in large agricultural areas. In this study, 16 2010 42310 3.0253 128000
days photosynthetic activity of pistachios was 2011 44097 2.5399 112000
investigated by integrating both field research 2012 53071 2.8264 150000
and modern techniques (remote sensing and 2013 54451 1.6272 88600
statistical methods). Furthermore, the effects of 2014 56186 1.4238 80000
climatic factors on the yield of pistachio were 2015 57996 2.4829 144000
analysed. Along with the present study, the 2016 60814 2.7954 170000
relationships between pistachio and wet and
dry climatic conditions were presented. It has
been observed that extreme rainfall conditions
have caused considerable changes on the
pistachio phenology and physiology. The area
harvested, yield and production of pistachio
between 2000-2016 years in Turkey are
represented Table 1 and Figure 2 (FAOSTAT,
2016). Pistachio yield and production values
fluctuate between years. Fluctuation or
irregular bearing are considered as
consequences of endogenous and exogenous
factors. Endogenous factors are defined as plant
own characteristics including cultivar,
agricultural practises, genetic and physiology
of plant (Nzima et al., 1997; Spann et al.,
2008). Exogenous or environmental factors
such as climatic variation of temperature and
precipitation, wet and dry climate condition are
significant predictor on yield (Elloumi et al., Figure 2. Changes Area harvested, Yield and Production
2013). of pistachio between 2000-20016 in Turkey
Middle Euphrates region provides a significant
portion of which Turkey pistachio production. Variation in climatic changes is of the
Turkey meets 20-25% of the world pistachio predictive factors on pistachio yield in Turkey.
production. Yield and yield per tree are low in In the study area, there has been extreme dry
Turkey. Kumar et al. (2016) reported that the period (severe dry, very severe drought,
extraordinary dry) 5 times in last 54 years. In
161
contrast to this, it is observed 8 times extreme or external-induced perturbations result
humid period (very humid, extremely humid, changes in biochemical and physiological
extraordinary humid). The year 2014 is the aspects as a consequence of molecular level
most arid period of the last 54 years in the changes. As a response to drought conditions,
study area. 2012 is one of the five most humid Khoyerdi et al. (2016) reported the shifts in
periods of the last 54 years (Figures 3, 4). value of vegetative growth, RWC, TChl and
Pistachio nut yield changes depending on dry carotenoids, TSP and an increase in MDA and
and wet periods in Gaziantep affect the average H2O2 etc. while reduction rate concerned with
yield of Turkey. Accordingly, the year 2014 is cell expansion and division rate were reported
an extraordinary dry period in the by Li et al. (2009).
meteorological station of Gaziantep, Şanlıurfa Based on the literature, we cannot deduce or
and Adıyaman. Turkey's average yield was simplify the factors responsible for irregular
1.4238 t/ha of pistachios for 2014. In contrast, bearing, which is of the complex phenomenon
the average value of Turkey was found to be of the plant kingdom (Spann et al., 2009).
2.8264 t/ha for 2012 when is excessive wet
period. CONCLUSIONS
Pistachio trees require very hot climates in
summers and cold climates in winters. In this Extreme conditions in the climate play an
context, pistachios are resistant to extreme hot important role on the crop production.
conditions, while photosynthetic activity Meteorological drought is one of these extreme
declines during the dry periods. It is observed climate conditions. Depending on global
that the photosynthetic activity of pistachio is climate changes, plant growth and physiology
particularly low in the growing season during are influenced and causes many problem in
the dry season of 2014. In contrast, the agriculture. According to the results of the
Enhanced Vegetation Index (EVI) values are drought analysis of Gaziantep, Sanliurfa and
above normal in the corresponding humid Adiyaman (in the south-eastern parts of
period of 2012 (Figure 5). Turkey) stations, pistachio is significantly
affected by extreme climatic conditions. The
Wet Period photosynthetic activity of pistachio trees grown
3,50 in Gaziantep, Sanliurfa and Adiyaman
increased during the periods of wet climatic
3,00 conditions. Especially at the beginning of 2010,
Dry Period pistachio plant exhibited highest photosynthetic
2,50
Yield (tonnes/ha)
activity and the activity was considered to be

2,00 associated with wet climatic conditions in both
areas at the end of 2009. The present results
1,50
indicate that the pistachio does not react
1,00 immediately to wet climatic conditions,
suggesting that pistachio respond or develop
0,50 any mechanisms to this climatic stimulant 2
0,00 and 3 months after the humid climatic
conditions.
It has been observed that pistachio was affected
Years
by extreme climatic conditions and extreme
rainfall conditions have caused considerable
changes on the pistachio phenology and
Figure 3. Yield of pistachios in wet and dry periods physiology. As a result it was observed that the
photosynthetic activity of pistachio increased
As reported in many and various studies, any during the periods of wet climatic conditions.
shift in metabolic activities of plants which
might be consequences of plant-own structure
162
Figure 4. Drought analysis results of Standardized Precipitation Index (SPI) of the Adıyaman (A), Gaziantep (B) and
Sanliurfa (C) (1952-2018) Source: General Directorate of Meteorology (MGM)
Figure 5. Photosynthetic activity in wet and dry periods according to EVI analysis results
163
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315-327. Khoyerdi F.F., Shamshiri M.H., Estaji A., 2016. Changes
Box E., Holben B.N., Kalb V., 1989. Accuracy of in some physiological and osmotic parameters of
AVHRR vegetation index as a predictor of biomass, several pistachio genotypes under drought stress.
primary productivity and net CO2 flux. Vegetation, Scientia Horticulturae, 198: 44-51.
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Çelik M.A., Sönmez M.E., 2013. Kızıltepe İlçesinin Köylerinde Antepfıstığı Üretim Maliyetlerinin
Tarımsal Yapısındaki Değişimlerin MODIS NDVI Karşılaştırılması. Uludağ Üniversitesi Ziraat
Verileri Kullanılarak İzlenmesi ve İncelenmesi, Fakültesi Dergisi, 25 (1): 41-51.
Marmara Coğrafya Dergisi, Sayı: 27: 262-281. Kumar P., Sharma S.K., Chandel R.S., Singh J., Kumar
Cheng Q., Wu X., 2011. Mapping Paddy Rice Yield in A., 2016. Nutrient dynamics in pistachios (Pistacia
Zhejiang Province Using MODIS Spectral Index. vera L.): The effect of mode of nutrient supply on
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Chmielewski F.M., Rötzer T., 2001. Response of tree temperate ecosystem. Scientia Horticulturae, 210:
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Agricultural and Forest Meteorology, 108: 101-112 Li Y., Sperry J.S., Shao M., 2009. Hydraulic
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164
LOW COST AND LABORATORY SCALE NIR SPECTROSCOPY

FOR QUALITY EVALUATION OF FRUITS AND VEGETABLES
Zehan KESİLMİŞ1, Kubilay K. VURSAVUŞ2

1
Adana Science and Technology University, Department of Electrical and Electronics Engineering,
Adana, Turkey
2
Çukurova University, Department of Agricultural Machinery and Technologies Engineering,
Adana, Turkey
Abstract
NIR spectroscopy has proved to be one of the efficient and easy tools to monitor the quality of agricultural products.
NIR spectrometers are versatile devices to monitor the ripeness or quality parameters of the fruits. We demonstrate a
low-cost spectrometer design that is produced with off the shelf components. In this work, the development,
characterization and validation of a prototype is discussed. The proposed device has a dedicated user interface on the
PC to plot and analyze spectral data. The performance of the proposed spectrometer is comparable to existing
laboratory scale spectrometers in terms of stability and resolution. The spectral resolution and response range of the
proposed spectrometer are 20 nm and 640-1050nm, respectively. Proposed device consists of MEMS based Hamamatsu
spectrometer sensor (C11708MA), microcontroller (Arduino) and IR light source. Roles of the Arduino are generating
essential control signals and sampling output of the C11708MA. These spectral response data have a huge advantage
in generating data sets that may be useful in building machine learning based models.
Key words: fruit and vegetable quality spectrometer, near infrared spectroscopy, non-destructive detection.
INTRODUCTION design that is produced with off the shelf

components.
An NIR spectrometer consists of a light source
Spectroscopy is gaining importance for fruit
(tungsten, halogen or LED), sample
firmness detection, pharmaceutical and
presentation accessory, detector, and optical
environmental monitoring. Near infrared (NIR)
components, such as lenses. NIR radiation
is a fast and non-destructive method that can be
covers the range of the electromagnetic
applicable to any biological or non-biological
spectrum between 780 and 2500 nm. NIR
materials. NIR technique has been widely used
spectrometer is a device that irradiates the
to measure internal qualities of various fruits
sample with NIR radiation and measures the
such as tomato, apple, and kiwifruit (Huang,
reflected or transmitted radiation.
2018; Ye et al., 2017; Li et al., 2017).
There are three different measurement setups
Spectroscopy technique can be utilized in
for obtaining near infrared spectra. These are
detecting plant diseases (Khaled et al., 2018).
reflectance, transmittance and interactance
Besides, multiple spectrometer modules can be
modes. In reflectance mode light source and
installed into packaging or processing lines for
sensor are mounted under a specific angle. In
measuring quality characteristics of the goods.
transmittance mode the light source is
However, most spectrometer equipment which
is utilized in the industry or laboratory is positioned opposite to the sensor. In
interactance mode the light source and detector
expensive and bulky. Although, there has been
are positioned parallel to each other.
some work on portable and low cost
Micro Electro Mechanical Systems (MEMS)
spectroscopy devices, there is a need for
offers inherently device miniaturization and
several improvements in terms of quality and
wide applications in sensors and actuators
repeatability of data (Das et al., 2016). Also,
Thanks to MEMS technology spectrometer
excessive price of these devices makes them
components can be easily produced in one
less accessible for many researchers. This work
package. Proposed spectrometer utilized with
aims to demonstrate a low-cost spectrometer
165
MEMS based Hamamatsu spectrometer sensor MATERIALS AND METHODS
(C11708MA) (Hamamatsu, 2018). C11708MA
spectrometer sensor allows designing low price In this section, the essential components of the
and high-performance spectrometer design. proposed spectrometer are described in detail.
Longitudinal section view of the C11708MA is The block diagram of the spectrometer system
given is Figure 1. designed in this study is given in Figure 2.
Fibre optics
USB Embedded
Light
electronic
Source
system
Sample Holder
Computer
Figure 2. System containing computer,

embedded system and light source
Figure 1. Longitudinal section view Proposed device consists of three parts:

of the C11708MA (Hamamatsu, 2018) computer, embedded electronic system and
light source. Light source holds the NIR lamp
C11708MA is equipped with 256-pixel CMOS and fibre optic cable in an appropriate
linear image sensor, micro optical slit (75 x 750 geometry. In our work, we prefer tungsten lamp
µm), a grating that is formed on a convex lens. as the source of the NIR light. Higher light
These features have made the C11708MA very intensity levels can be obtained by using
compact and versatile. tungsten lamp. Power consumption of tungsten
In this work, we demonstrate a low-cost lamp is somewhat high respect to the LED and
spectrometer design that is produced with off laser sources. In future works for the mobile
the shelf components. and battery-based operation system can easily
The development, characterization and utilize with LED light source.
validation of a prototype are discussed. The Embedded electronic system is responsible for
proposed device has a dedicated user interface the generating control signal of the MEMS
on the PC to plot and analyse spectral data. spectrometer chip.
The performance of the proposed spectrometer The block diagram of the embedded electronic
is comparable to existing laboratory scale system is given in Figure 3.
spectrometers in terms of stability and
resolution. Technical specifications of the MCP6001
C11708MA are given in Table 1.

USB
Table 1. Technical specifications of the C11708MA 5V

Ground
Dimensions 27.6 x 16.8 x 13 mm

Video
Weight 9g EOS
Gain
Start
Spectral range 640-1050 nm Clock
C11708MA 74LS125 Arduino UNO

Supply Voltage 5V
Power Consumption 30 mW
Figure 3. Block diagram of the embedded
electronic system
Number of pixels 256
Arduino UNO generates clock pulse for

The digital resolution and response range of the C11708MA which was set to 1 kHz in the
proposed spectrometer are 2 nm/pixel and 640- proposed design. Arduino sends a start pulse to
1050 nm, respectively. The designed device C11708MA to trigger the pixel read-out
and supplied software is user friendly so that process and the interval between two start
anybody with basic knowledge of spectroscopy pulses is accepted as the integration time of the
can easily use it. Spectrometer device sensor. When Arduino receives a capture
communicate with computer via USB cable. command from MATLAB, a start pulse was
166
triggered which initiated the charge integration Off the shelf fibre optic cable has been used for
of each pixel. The C11708MA provided an End the assessment of light levels inside the fruit
of Scan (EOS) pin which was used to terminate sample which has been illuminated with NIR
the read out process. As declared above, main light source. The probe has minimal effect on
purpose of the embedded electronic system is the light being measured and enables the direct
sampling and sending the spectrum data to the measurement of light levels at any location in
computer. Embedded electronic system is the fruit.
controlled by the computer software for The spectrometer, light source and sample
coherent detection of spectrum. Embedded holder design was wholly completed in
electronic system utilized with low cost Sketchup (Sketchup, 2018), a solid modelling
Arduino microcontroller board. Roles of the software designed by Google. The design of the
Arduino are generating essential control signals case body is shown schematically in Figure 4.
and sampling output of the C11708MA. The case was printed with polylactic acid
Arduino UNO contains built in 10-bit precision (PLA) media.
Analog to Digital converter (ADC). Resolution
of this ADC is suitable for NIR spectrometer Sample holder Light Out
application. C11708MA contains 5 digital I/O

interface pins for logical control and one analog
output for spectral response data. To avoid Light in
excessive current sink from analog output port

buffered with OPAMP (MCP6001) buffer
amplifier. Fibre Optic
Cable
The embedded software based upon the open- Spectrometer Case Light Source
source Arduino IDE platform. The Arduino

firmware was developed with the Arduino IDE Figure 4. 3D design of the spectrometer
and provides an easy to navigate hierarchical
menu system for selection of device functions. RESULTS AND DISCUSSIONS
The computer software was developed in the
MATLAB environment. The computer The spectra of apple and carrot fruit samples
software stores and displays received spectrum are shown in Figures 5, 6, respectively. The
information. similar spectral pattern was also observed in the
As decelerated above, C11708MA contains 256 spectra of apple fruit measured in the
pixels to detect specific wavelengths. These interactance mode.
pixels are numbered and these numbers have to
be transformed into the corresponding
wavelength. This can be done by a multiorder
polynomial as given in Eq. (1).
(nm) A0 B1x B2 x2 B3 x3 B4 x4 B5 x5 (1)
where, x is the pixel number and A0, B1, B2, B3,

B4 and B5 are calibration coefficients whose
values are supplied by the sensor producer.
This approximation polynomial gives a
precision wavelength vector. Computer
software generates a wavelength vector by
using Eq. (1). Computer software also detects Figure 5. Transmittance spectrum of apple
the peak and Full Width at Half Maximum
(FWHM) values of the spectrum. Peak and
FWHM values are important for evaluating the
properties of the sample.
167
ACKNOWLEDGEMENTS
Authors would like to thank Mr. Kayhan

KESİLMİŞ for valuable support for 3D design
and production of spectrometer case. Authors
also would like to thank to Erdal Machine
Limited Company for its valuable contributions
in this research.
REFERENCES
Das A.J., Wahi A., Kothari I., Raskar R., 2016. Ultra-
portable, wireless smartphone spectrometer for rapid,
non-destructive testing of fruit ripeness. Scientific
reports, 6, 32504.
Figure 6. Transmittance spectrum of carrot Hamamatsu web site accessed: 23.02.2018
https://www.hamamatsu.com/jp/en/C11708MA.html
Huang Y., Lu R., Chen K., 2018. Prediction of firmness
CONCLUSIONS parameters of tomatoes by portable visible and near-
infrared spectroscopy. Journal of Food Engineering,
In conclusion, the designed MEMS based NIR 222, 185-198.
spectrometer is a portable and low-cost device. Khaled A.Y., Abd Aziz S., Bejo S.K., Nawi N.M.,
In the Results and Discussions section it was Seman I.A., Onwude D.I., 2018. Early detection of
diseases in plant tissue using spectroscopy-
shown that proposed device works with applications and limitations. Applied Spectroscopy
sensitivity and resolution acceptable for many Reviews, 53 (1), 36-64.
tasks both in education and research. Future Li M., Pullanagari R.R., Pranamornkith T., Yule I.J.,
work is necessary for this design to realize the East A.R., 2017. Quantitative prediction of post
full potential of the tool for fruit internal storage ‘Hayward’ kiwifruit attributes using at
harvest Vis-NIR spectroscopy. Journal of Food
properties detection applications. As discussed Engineering, 202, 46-55.
above main role of this device is obtaining set Sketchup web site accessed: 23.02.2018
of spectral response data for various types of https://www.sketchup.com/
fruits. These spectral response data may be Ye X., Ishioka S., Zhang S., 2017. Estimation of the
useful in building machine learning based degree of red coloration in flesh of a red-fleshed
apple cultivar ‘Kurenai no Yume’ with a UV-vis-NIR
models. In addition, work is needed to make interactance device. Post-harvest Biology and
the NIR spectrometer portable, such as the Technology, 124, 128-136.
incorporating batteries.
168
INCREASING THE ECONOMIC PERFORMANCE BY PROMOTING
HIGH-DENSITY APPLE ORCHARDS IN THE DÂMBOVIŢA FRUIT BASIN
Gheorghe PETRE1, Valeria PETRE1, Adrian ASĂNICĂ2

1
Research and Development Station for Fruit Growing Voineşti, 387 Main Street,
Voineşti, Romania
2
Abstract
The researches carried out at the Research and Development Station for Fruit Growing Voineşti in the period 2015 -
2017, had as objective the evidence of modern plantations established after 2007 by the producers of SC Mere de
Voineşti SRL; Owner Dan Ionescu, and I.I. Luminita Marin from Malu cu Flori. The tree culture in high density system
extends mainly to apple trees in the Dâmboviţa fruit-tree basin, the interest of fruit growers is evident in the promotion
of productive varieties, well adapted to the pedoclimatic conditions of the area, grafted on M.9 rootstock planted at a
distance of 3,5 x 1 m, with a density of 2,857 trees/ha. The biggest yield belonging to SC Mere de Voineşti SRL were
harvested from following varieties: ʻGolden deliciousʼ (43.2 t/ha), ʻBraeburnʼ (35.4 t/ha) and ʻGalaʼ (35.2 t/ha), trees
that are 9-11 years old, in comparison with ʻGalaʼ apple varieties (36.2 t/ha) and ʻIdaredʼ (33.8 t/ha) of 7-8 years old.
In the apple tree orchard for 3 years, in the ʻGolden deliciousʼ and ʻIonaprinceʼ varieties, production of 40.2 t/ha and
38.6 t/ha was achieved in 2017. At the Owner Dan Ionescu, with 8 years old trees, the most productive varieties were:
ʻGolden deliciousʼ (38.9 t/ha), ʻIdaredʼ (34.7 t/ha) and ʻGalaʼ (32.7 t/ha), and the producer I.I. Luminita Marin ,
located in Malu cu Flori, with trees in the 4th year after planting, recorded 34.3 t/ha in the ʻRed Galaʼ variety, 46.8
t/ha in ʻGolden deliciousʼ and 42.8 t/ha to ʻPinovaʼ. The high density apple system is recommended for expansion in the
well-established fruit-growing areas of our country, including the Dâmboviţa fruit basin, due to the high yield and
efficiency, the way of periodic and rapid replacement of the assortments, demanded more and more by consumers.
Key words: high apple density system, apple assortment, yield, crop systems.
INTRODUCTION full economic efficiency. The generalization of

the high-density apple system (Petre Gh. et al.,
Fruit growing remains one of the field of 2009), represents the upgrade of the moder-
horticulture of great interest to growers in the nization of the fruit growing sector in our
traditional fruit-growing areas in Romania. country, including the Dâmboviţa basin.
Both from an economic and social point of
view, the cultivation of trees and in particular MATERIALS AND METHODS
the apple trees provides permanent activity and
manages to capitalize inappropriate areas for By implementing the fruit tree thematic
other agricultural crops. subprogram of PNDR in the period 2015 -
The extension of modern apple culture systems 2020, the aim is to increase the technical and
with higher precocity and short-term economic competitiveness in fruit growing
exploitation is a way of periodic and rapid sector, by promoting technologies adapted to
replacement of the assortments (Petre Gh., the pedoclimatic conditions in Romania, with
2006). In this regard, introduction of modern the goal to set up new apple plantations,
techniques and novelties in the production including in the Dâmboviţa fruit basin.
adapt better the apple to the high quality In order to emphesize the interest of the
European standards (Comanescu D., 2015; Dâmboviţa fruit growers in the promotion of
Petre Gh. et al., 2005). modern culture systems, during the period
Apples are nowadays cultivated in high density 2015-2017 some high-density apple
system and faster expand in Romanian fruit plantations, established after 2007, by some
growing areas, the results achieved confirm its apple producers from Dâmboviţa fruit basin
169
were highlighted: SC Mere de Voineşti SRL; high density culture system, the interest of the
Owner Dan Ionescu; I.I. Luminita Marin from fruit growers is evident through the establish-
Malu cu Flori. ment of new modern plantations, using
Data on the planted area, varieties and European varieties, proved to be well adapted
rootstocks used, planting distances, soil mainte- to the pedoclimatic of the county.
nance, crown shape, phytosanitary treatments
scheme, fertilization etc. were recorded. Also, Evidence of high-density apple systems in
the production potential of the apple trees was the Dâmboviţa fruit basin
monitored, especially since the newly The significant successes achieved in the world
established plantations used trees from Italy or fruit tree, but especially in the European one in
Netherland, with varieties different from the recent years, as well as the valuable experience
ones regulary cultivated in Romania. gained, led some of the farmers in the
Dâmboviţa fruit basin to show interest in
RESULTS AND DISCUSSIONS expanding the apple-tree system, replacing the
old plantations with over aged trees charac-
The high density apple system offers the terized by a low production potential.
possibility of easy change the assortment (due The tendency to expand the high-density
to the short period of cultivation), pedestrian apple orchards to some of the Dâmboviţa fruit
labour in the orchard, increased performance in growers is outlined in Table 1 by planted
terms of yield and production quality. areas, the age of trees and the density of
In recent years, in the Dâmboviţa fruit tree planting.
basin, especially apple tends to generalize the
Table 1. The examples of high density apple orchards in the Dâmboviţa County
Area planted Year of Trees age Plantng
No Owner Trees/ha
(ha) planting (years) distances (m)
1 SC Mere de Voineşti SRL 4 2007 11 3.5 x 1 2,857
4 2009 9 3.5 x 1 2,857
2 2010 8 3.5 x 1 2,857
7 2011 7 3.5 x 1 2,857
6 2015 3 3.5 x 1 2,857
2 Dan Ionescu 5 2010 8 3.5 x 1 2,857
3 I.I. Luminita Marin -
1.50 2014 4 3.5 x 1 2,857
Malu cu Flori
From the data presented in the above table, SC grafted on M9 rootstock. The fruit trees were
Mere de Voineşti SRL has an area of 23 ha, purchased from Italy.
cultivated with apple in a high density system, with The area of 5 ha cultivated with apple by Dan
trees aged from 3 to 11 years, planted at a distance Ionescu, the assortment consists of the apple
of 3.5 x 1 m, of 2,857 trees/ha. At the same planting varieties ‘Golden delicious’, ‘Idared’, ‘Granny
density (2,857 trees/ha), but with the age of trees of Smith’, ‘Braeburn’, ‘Gala’ grafted on the M9
4-8 years old, there are apple orchards belonging to
rootstock. The Dutch system is present with a
Dan Ionescu farmer, with a surface of 5 ha and I.I.
Luminita Marin - Malu cu Flori with a surface of
wood tutor for each individual tree.
1.5 ha. The irrigation system is layered on the ground
The 23 ha, cultivated with apple at SC Mere de and provide a flow rate of 2 l/h.
Voineşti SRL, are equipped with a 3-wire trellising At the I.I. Luminita Marin from Malu cu Flori,
system, the first one placed at 60 cm from the assortment consists of ‘Pinova’, ‘Red Gala’
the ground, on which the drip irrigation system of and ‘Golden delicious’ varieties grafted on M9.
2.2 l/h is tied up. The support system has 2 wires, the first wire is
The irrigation system can be used for fertilizers. sitated at 60 cm from the ground level, on
The apple assortment consists of varieties which the drip irrigation pipes are installed and
‘Braeburn’, ‘Gala’, ‘Ionaprince’, ‘Granny Smith’, work with a flow rate of 2 l/h. Each tree is
‘Golden delicious’, ‘Stark’ group of different types
supported by a bamboo stick.
170
Apples cultivated on these plots, are susceptible Evaluation of production potential in apple-
to diseases. tree orchards in the Dâmboviţa fruit basin
Therefore, to ensure a proper phytosanitary In 2017, due to the low temperatures below the
state, a number of 16-18 phytosanitary freezing threshold recorded in the area, the
treatments have been applied annually. yields were substantially reduced depending on
In the overproduction years, the chemical the variety and age of the trees.
thinning of the fruit, followed by a manual thin, Analyzing the productions obtained by SC
was applied to obtain fruits with valuable Mere de Voineşti SRL during the 3 years of
appearance and price on the market. study (2015-2017), it is obvious that the most
In all of these apple orchards, the trees are productive apple varieties in the 9-11 years old
trained as Spindle bush and the soil is trees were ‘Golden delicious’ with 43.2 t/ha,
maintained between the rows covered with followed by the ‘Braeburn’ apple variety with
grass and clean within the row using different 35.4 t/ha and ‘Gala’ with 35.2 t/ha (Table 2).
herbicides.
Table 2. The apple yield obtained at the varieties cultivated in the high density system between 2015-2017 by
SC Mere de Voineşti SRL
Apple trees Trees/ Variety/M9 Yield (t/ha), year Average
age (years) ha 2015 2016 2017 (t/ha)
9 - 11 2,857 ʻGalaʼ 42.0 47.1 16.5 35.2
ʻGolden deliciousʼ 36.0 75.1 18.6 43.2
ʻBraeburnʼ 48.0 50.0 8.3 35.4
ʻStark deliciousʼ 20.0 45.1 15.0 26.7
ʻGranny Smithʼ 21.0 41.7 12.0 24.9
7-8 2,857 ʻGalaʼ 36.8 53.1 18.6 36.2
ʻIonaprinceʼ 20.4 45.1 15.2 26.9
ʻIdaredʼ 35.6 47.1 18.6 33.8
3 2,857 ʻGolden deliciousʼ 18.2 14.6 40.2 24.3
ʻIonaprinceʼ 12.6 15.7 38.6 22.3
In the 7-8 years old trees, the highest yields, Dan Ionescu, in 2017, in 8-year-old trees,
averaging 3 years of production, were recorded registered a drastically reduced production
by ‘Gala’ variety with 36.2 t/ha, followed by compared to the previous year due to the frost
the ‘Idared’ variety with 33.8 t/ha. In the apple phenomena. The smallest production was
orchard with 3 years old trees, in 2017, the recorded in the ‘Braeburn’ variety by 8 t/ha
‘Golden delicious’ variety, recorded a (considered less adapted to the climatic
production of 40.2 t/ha and 38.6 t/ha for the conditions in the Voineşti area). The highest
‘Ionaprince’ variety. The production was yields, were obtained by ‘Golden Delicious’
considered as normal because the flower buds with 31.3 t/ha, followed by ‘Idared’, with 20.6
in the frozen moment was in the balloon stage t/ha (Table 3).
and were not affected.
Table 3. The apple yield obtained at the vaieties cultivated in the high density system between 2015-2017 by
Dan Ionescu and I.I. Luminita Marin - Malu cu Flori
Apple trees Trees/ Variety/M9 Yield (t/ha), year Average
age (years) ha 2015 2016 2017 (t/ha)
8 2,857 ʻGolden deliciousʼ 25.0 60.3 31.3 38.9
ʻIdaredʼ 18.3 65.1 20.6 34.7
ʻGranny Smithʼ 18.2 30.3 15.5 21.3
ʻBraeburnʼ 24.3 59.4 8.0 30.6
ʻGalaʼ 24.8 58.8 14.6 32.7
4 2,857 ʻPinovaʼ 30.0 35.7 42.8 36.2
ʻRed Galaʼ 30.0 25.1 34.3 29.8
ʻGolden deliciousʼ 29.8 30.0 46.8 35.5
171
Analyzing the production recorded over the 3
years of study, the highest yields were recorded
in the ‘Golden delicious’ apple variety with
38.9 t/ha, ‘Idared’ by 34.7 t/ha and ‘Gala’ with
32.7 t/ha.
Luminita Marin, located in Malu cu Flori,
harvested from the 4th year trees, in 2017 34.3
t/ha to 46.8 t/ha, depending on variety, as
follows: ‘Red Gala’ - 34.3 t/ha, ‘Golden
delicious’ 46.8 t/ha and ‘Pinova’ 42.8 t/ha.
Analyzing the productions recorded during the Figure 1. The efficiency and the exploitation period of
3 years of study, ‘Pinova’ was the most apple orchards in different cultural systems
productive apple variety with 36.2 t/ha close to
next one ‘Golden delicious’ with 35.5 t/ha. The differences between the apple culture
systems are quite obvious in terms of fruit input
The efficiency of the high-density apple of the trees, the economic growth and the level
system compared to other cultivation of production during the full production period,
systems as well as the upper limit of the exploitation
The intensification of the fruit growing sector period.
in our country must accept as an important What really differentiates apple cultivation
objective the introduction of new intensive crop systems is the precocity. The classic system
cultivation systems adapted to the new socio - start be economically after 10-12 years from
economic conditions and to the continuously planting, to the intensive system after 6-7 years,
developing technical and material basis. and to the high density after 3-4 years.
In connection with the introduction of new crop If we take into consideration the first 10 years
systems, it should be noted that the conven- since planting, it is clear that the yields
tional or classic culture system has a number of produced significantly differences in the apple
limits beyond which it is not overpassed, culture systems. If the high-density and even
regardless of the applied technologies. intensive system yields/ha is high as a result of
The production of fruit is delayed in the the rapid entry of the fruit trees and the
classical orchards, from 8-10 years onwards achievement of a large productive volume of
after planting. Some high-volume main works, the crown through the planting density itself,
such as pruning and harvesting fruit, require the classic system should expect much more for
excessive workload due to the high tree height, both the bearing of fruit trees and the formation
7-10 m, which forces growers to use large and of the skeleton branches for yield bearing.
inconvenient stairs. The mechanized execution The high density apple system is recommended
of phytosanitary treatments, soil mainenance is for extension in the well-established fruit-
hampered by the globular form of the crown or growing areas of our country, including the
is realised with low efficiency and unecono- Dâmboviţa fruit basin, due to the high yield and
mically. efficiency, the way of periodic and rapid
Intensive crops and, above all, high-density replacement of the assortments, but also the
plants offer greater flexibility in changing fruit obtaining of bigger apples volumes demanded
varieties due to the lower economic exploi- more and more by consumers.
tation period of these orchards (22 to 30 years
in intensive orchards, 15-16 years to high CONCLUSIONS
density ones).
The high density system expands mainly to
The yields of the high density production of
apple trees in the Dâmboviţa fruit basin, the
apple during the exploitation period are
interest of fruit growers is evident in the
superior, reflected by the productive potential
promotion of productive varieties well adapted
differences compared to the intensive and
to the pedoclimatic conditions of the area,
classic culture systems (Figure 1).
grafted on the M9 rootstock planted at a
172
distance of 3.5 x 1 m, with planting density of Luminita Marin from Malu cu Flori, highlight
2,857 trees/ha. the 4th year trees, that produced from 34.3 t/ha
Although the production was diminished by the to 46.8 t/ha It was remarked ‘Red Gala’,
low temperatures recorded during the flowering ‘Golden delicious’ and ‘Pinova’.
period in 2017, the highest average production
for three years period belonging to SC Mere de REFERENCES
Voineşti was recorded in 9-11 years old apple
trees by the varieties: ‘Golden delicious’, Comănescu D.N., 2012. Cercetări privind sistemul de
mare densitate la măr, în scopul obţinerii de producţii
‘Braeburn’ and ‘Gala’ and in 7-8 years old adaptate la cerinţele de comercializare, Teză de
apple trees, the varieties ‘Gala’ and ‘Idared’. doctorat.
In the apple tree orchard for 3 years, the Petre Gh., 2006. Cerinţele pomiculturii moderne. Revista
‘Golden delicious’ and ‘Ionaprince’ varieties Amsem, nr.7, decembrie.
produced 40.2 t/ha and 38.6 t/ha in 2017. Petre Gh, Andreieş N, Petre V., 2005. Tehnologia
obţinerii unor producţii de mere competitive. Editura
In Dan Ionescu orchard, the 8 years old trees Pilner Târgovişte.
performed better in varieties such as: ‘Golden Petre Gh, Petre V., Comănescu D.N., 2009. Promovarea
delicious’, ‘Idared’ and ‘Gala’. sistemelor moderne la măr, în pomicultura
românescă. Revista Agricultorul român, nr.9.
173
174
TWENTY YEARS OF EXPERIENCE IN INTENSIVE PLUM

PRODUCTION ON PRUNUS CERASIFERA EHRH. ROOTSTOCK
Miljan CVETKOVIĆ1, Nikola MICIC1, 2

1
University of Banja Luka, Faculty of Agriculture, Bulevar Vojvode Petra Bojovića 1A,
78000 Banja Luka, Bosnia and Herzegovina
2
University of Banja Luka, Genetic Resources Institute, Bulevar Vojvode
Petra Bojovića 1A, 78000 Banja Luka, Bosnia and Herzegovina
Abstract
As a result of two decades long research this paper provides basic parameters and characteristics of training system
plum spindle on Myrobalan (Prunus cerasifera Ehrh.) seedling rootstock. Utilization of Myrobalan rootstock induces
intensive plum growth in initial years of production. This growth can only be controlled by application of adequate and
precise pomotechnics and induction of early fruiting as a result of applied treatments. Obtained results indicate that
training principles for plum spindle system are similar for most analyzed cultivars. On the other hand, cultivar specifics
concerning biological predispositions for this training system may greatly influence success of production. Certain
cultivars (ʻČačanska Lepoticaʼ and ʻStanleyʼ) can serve as model plants for correct and successful high intensity plum
production in spindle system. All factors should be taken into consideration, when defining training systems adequate
for plum (Prunus domestica L.) on Myrobalan rootstock, so the optimal combination could be defined depending on
growing conditions and level of application of agrotechnical measures. Level of applied measures in orchard needs to
be one of the most important elements of financial feasibility of plum production in spindle system. Planting density
depending on the cultivar and growing conditions is between 1000 and 2000 trees per hectare. Increase in number of
individual spindles per hectare is possible by modification of planting system (“V” spindle planting) or modification of
training system (multi-leader spindle systems), which implies different approach to production.
Key words: canopy management practices, cultivar, density.
PLUM PRODUCTION ON MYROBALAN suppression and initiation of greater generative

SEEDLING ROOTSTOCK bud differentiation. In pome fruits, this results
in enhanced fruiting potential, and the fact that
The introduction of dwarfing rootstocks in fruit pome fruit generative buds contain vegetative
production has led to its intensification, cones means that an increase in the total
primarily through changes in orchard systems number of generative buds per tree does not
and increases in planting density and yield per induce a decrease in leaf surface area per fruit
unit area (Hrotko et al., 1998; Achim et al., i.e. there is no decrease in the total number of
2017). Apart from significant initial leaves required for successful fruit
investments, vegetative rootstocks of weak development in trees bearing a high crop load
vigor require optimum growing conditions and (Lučić et al., 1996). On the other hand, in stone
proper cultural and tree management practices fruits, growth suppression and increased
to manifest their positive characteristics. generative bud differentiation lead to a
Impressive results in improving orchard decrease in vegetative buds per tree and, hence,
systems for apple and pear have given rise to leaf biomass per fruit. There is the impression
intensive research in dwarfing rootstock that intensive production systems for stone
selection for stone fruits as well. However, fruits can be successfully established on semi-
intensive systems of production on dwarfing dwarfing rootstocks using specific canopy
rootstocks have not been as successful for stone management practices that enable fruiting each
fruits as for pome fruits (Paunović et al., 2011). year. Regardless of the development of plum
This can be explained by the fact that dwarfing vegetative rootstocks, the Myrobalan Prunus
rootstocks modify scion growth and cerasifera Ehrh. seedling is still a predominant
development through physiological growth rootstock in the Bosnia and Herzegovina and
175
Serbia, and some other Balkan countries as During the period, it is necessary to use all tree
well (Botu and Botu, 2017). Less favorable soil management treatments which, apart from
conditions for plums grown in this area, partial having a role in tree architecture, ensure the
use of cultural and tree management practices fastest possible formation of fruiting wood and
in orchards, higher costs of vegetative fruit-bearing branches, as the best procedure for
rootstocks and the resulting nursery trees, as initial vigor suppression. Successful plum
well as the marked presence of the traditional production is only possible if intensive summer
approach to plum production are just some of pruning operations are used (during the
the reasons for the current state. Plum is grown growing season), which is not the case in
on Myrobalan rootstock mostly in extensive or conventional training systems. To this end, all
semi-intensive orchards, with an average pruning treatments should be used at optimum
planting density of 400 to 800 trees per hectare dates. Notching is applied during dormancy or
(Micic et al., 2005; Botu and Botu, 2017), and immediately before the growing season (Lučić
using only minimum tree management et al.,1988; Glisic, 2012; Cvetković et al.,
practices (pruning during dormancy). 2017a). Plum has a biological predisposition to
Nevertheless, experience indicates that the creating “floors” i.e. branches growing in
intensification of plum production on several blocks along the central leader;
Myrobalan rootstock is possible and justified, if therefore, notching provides great results in
all necessary tree management practices are balanced targeted positioning of the shoots
used, especially in the initial years of along the central leader. Effects of notching are
production (Cvetkovic et al., 2017b). not related to its timing, and the cultivar-
Considering the high fruiting potential of dependent response to growing point initiation
currently grown plum cultivars, the specific ranges from 95.77-98.11% (Cvetković et al.,
manner of formation and character of fruiting 2017a). The formation of a crotch angle of
wood, as well as the pronounced tendency for approximately 90° relative to the central leader
decreased generative bud differentiation in provides the best predisposition to the creation
certain cultivars (regardless of rootstock type), of quality growth of main lateral branches and
strong vegetative growth of the Myrobalan fruit-bearing branches. In some cultivars
seedling rootstock can have a significant effect (‘Čačanska Lepotica’), this procedure is
on the regular formation and renewal of fruiting sufficient for the formation of main lateral
wood. This paper presents 20 years of branches properly positioned relative to the
experience in intensive plum production on central leader; however, this is not the case for
Myrobalan rootstock, specific characteristics of most cultivars. Spreading the branches and
the “plum spindle” training system, as well as maintaining their position at an initial crotch
the biological predisposition of certain cultivars angle of approximately 90° is usually
to this training system. performed with different types of wires and
twines (Mićić et al., 2005; Cvetković et al.,
CHARACTERISTICS OF MODERN 2017b), keeping in mind the timing.
“PLUM SPINDLE” TRAINING SYSTEMS Maintaining crotch angles by shoot twisting
must be applied successively during the
It is possible to train plum trees to a modified growing season before the newly formed shoots
spindle system, slightly higher (3.2-3.5 m) than are lignified at the base. Twisting is uniquely
the standard spindle, with a trunk height of up and highly effectively used in plum (Mićić et
to 0.5 m, and with more pronounced main al., 2005; Glisic, 2012; Cvetković et al.,
lateral branches at the base of the tree. The 2017b). Pruning during dormancy is applied in
branches should be distributed irregularly, accordance with standard principles and
spirally along the central leader, taking care cultivar specificity. Succession of old lateral
that their length is shortened towards the top so branches is usually done by “stub cuts” and
that the tree takes the final form of a “conical successive use of undercutting to initiate the
frustum” (Mićić et al., 2005). During the first 2 growth of lateral shoots which will take the role
or 3 years after planting, trees are trained of main lateral branches. Training plum trees to
according to the spindle training principles. a spindle system ensures satisfactory yields
176
(25-40 t/h), high quality fruit and ease of access production of ‘Čačanska Najbolja’ and
for fruit harvest, thus increasing the number of ‘Čačanska Rana’ is strong initial vigor and self-
extra quality fruits in the total production sterility issues which very often additionally
structure. contribute to strong vigor. ‘Katinka’ requires
more precise pruning combined with some
CULTIVAR SPECIFICITY AND other measures to achieve the satisfactory fruit
SUITABILITY FOR THE “PLUM size.
SPINDLE” TRAINING SYSTEM Table 1. Biological specificity of growth and
development that impede spindle training system
The biological specificity regarding the growth Cultivar Specificity
and development of individual cultivars, as ‘Čačanska vigorous
well as their response to tree management Najbolja’ self-sterile
late first bearing
treatments are of special importance for very intensive shoot growth in initial years of
defining a proper approach to training certain production
cultivars to the spindle system. Morphological insufficient number of strong shoots adequate for
main lateral branches
characteristics of growth and development non-uniform lateral branching of the central
largely affect the formation of the “plum leader
spindle” training system and high-density acute crotch angles of the lateral branches relative
to the central leader
planting. Tree architecture, type of growth of lack of response to treatments inducing the
fruit-bearing branches, intensity of formation succession of main lateral branches
and character of fruiting wood, dominant type ‘Elena’ vigorous
of fruit-bearing branches, fruit positioning on very intensive shoot growth in initial years of
production
the tree, degree of fruit-bearing branch long shoots
shedding and moving further from the central extremely acute crotch angles of lateral branches
leader after fruiting and the specific activation relative to the central leader
fast shoot lignification at the base
of new points of growth for the scheduled highly intensive transfer of growth points to top
succession of main lateral branches are just parts of the tree
some of the cultivar-specific morphological unfavorable response to the succession of main
lateral branches
characteristics of growth and development that ‘Čačanska self-sterile
greatly affect the spindle training approach Rana’ occurrence of alternate bearing
taken for individual cultivars. Certain late first bearing
shoots have specific growth dynamics
biological characteristics of cultivars (Table 1) insufficient number of strong shoots adequate for
can pose significant difficulty to the approach main lateral branches
taken to the formation and maintenance of the relatively unfavorable response to the succession
of main lateral branches
training system, which has to be taken into ‘Stanley’ intensive shoot growth in initial years of
consideration while designing the training production
system. Experience indicates that the above too high percentage of short fruit-bearing
branches in the total fruit-bearing branch structure
listed cultivars can be conditionally classified ‘Katinka’ alternate and partially alternate bearing
in three groups (Table 2), based on their tendency to form thin long fruit-bearing branches
biological predisposition to spindle training as dominant short fruit-bearing branches on main
lateral branches
well as on treatment intensity for the affinity towards shedding of the base of main
satisfactory formation and maintenance of the lateral branches
training system. ‘Čačanska Lepotica’ and transfer of growth points to top parts of the tree
‘Čačanska alternate bearing
‘Stanley’ are in the first group as model Rodna’ intensive growth of new shoots – longer treatment
cultivars for intensive production; their period
characteristics are suitable for spindle system long elastic shoots inappropriate for treatments
during the growing season
training. All treatments are highly effective and relatively unfavorable branching angles relative to
successful, even though they are somewhat the central leader
more extensive and demanding in ‘Stanley’. A extreme shedding of main lateral branches after
fruiting
more complex approach is necessary for intensive transfer of growth points to the
‘Čačanska Najbolja’, ‘Čačanska Rana’ and periphery of the tree
‘Katinka’. The greatest difficulty in the ‘Čačanska lack of formation of long main lateral branches
Lepotica’
177
‘Čačanska Rodna’ and ‘Elena’ belong to a necessity to move the intensification of plum
separate group as they are highly demanding as training systems on Myrobalan rootstock in
regards training to the spindle system and some other direction.
require high-intensity treatment regardless of
orchard age. CONCLUSIONS
Table 2. Cultivar predisposition to spindle training and Myrobalan (Prunus cerasifera Ehrh.) seedling
treatment intensity for proper training (more pluses is the dominant rootstock for plum in the
indicate better predisposition i.e. greater treatment
intensity)
Western Balkans. Even though it induces
stronger vigor, with adequate and timely use of
Cultivar Predisposition Treatment intensity
‘Čačanska Najbolja’ ++ +++ all necessary tree management treatments, it is
‘Elena’ + +++ possible to control growth and establish an
‘Čačanska Rana’ ++ ++
‘Stanley’ +++ +
appropriate balance of vegetative and
‘Katinka’ ++ ++ generative growth in initial years.
‘Čačanska Rodna’ + +++ Plums can be successfully grown in a modified
‘Čačanska Lepotica’ +++ +
spindle system, with a planting density of 900
PLANTING DENSITY to 2000 trees per hectare, which is largely
affected by cultivar-specific growth and
Planting density is dependent on the expression development.
of cultivar vigor in combination with Cultivars demonstrate significant differences in
myrobalan rootstock, as well as on cultivar their response to intensive tree management
specificity regarding tree architecture (Table 3). treatments, which should be considered when
designing an orchard.
Table 3. Planting density Tree management treatments during the
growing season are essential for plums to be
Cultivar Spacing Spacing Number of
between rows within the plants/ha trained to the spindle system.
(m) row (m) Treatments during dormancy should be
‘Čačanska 4.2 – 4.6 2.0 – 2.4 900 – 1190
Najbolja’ corrective and less severe so as to prevent
‘Elena’ 4.2 – 4.6 2.0 – 2.2 980 – 1190 additional vigor.
‘Čačanska Rana’ 4.0 – 4.4 1.8 – 2.0 1130 – 1380
‘Stanley’ 3.8 – 4.2 1.6 – 1.8 1320 – 1640 Plum production using this system involves
‘Katinka’ 3.8 – 4.0 1.6 – 1.8 1380 – 1640 more manual labor in the total cost structure,
‘Čačanska 3.8 – 4.2 1.4 – 1.6 1480 – 1870
Rodna’
but it ensures higher yields and optimal quality
‘Čačanska 3.6 – 4.0 1.4 – 1.6 1560 – 1980 of the fruit.
Lepotica’
REFERENCES
The highest spacing is required for cultivars
with strong vigor, which require more complex Achim G., Botu I., Botu M., Duţu I., Stănică F., Mazilu
tree management treatments. Experience shows C., 2017. The rootstok's role in modernizing plum
that ‘Stanley’ can be grown successfully at a culture. Acta Horticulturae, 1175: 49-54.
narrower spacing within the row (1.5 m), which Botu M., Botu I., 2017. Evaluation of intensive plum
culture system in the north of Oltenia - Romania.
is also true for ‘Čačanska Lepotica’ (1.2-1.4 Acta Horticulturae, 1175: 55-60.
m). The intensification of plum production Cvetković M., Mićić N., Đurić G., Bosančić B., 2017a.
using the spindle training system is additionally Leader management techniques to induce vegetative
possible through the “V” planting system bud development in plum. Acta Horticulturae, 1175:
(Mićić et al., 2005), which allows an increase 41-47.
Cvetkovic M., Djuric G., Micic N., 2017b. Canopy
in planting density by additional 25-30%, thus management practices in modern plum (Prunus
increasing crop yield (Mitrovic et al., 2005). domestica L.) production on vigorous rootstocks.
On the other hand, the need to use complex Scientific Papers. Series B, Horticulture. Vol. LXI,
trellises in this training system makes 117-122.
investment more expensive. At the same time, Glišić I., 2012. Pomotehničke specifičnosti šljive gajene
u gustoj sadnji. Doktorska disertacija. Agronomski
difficulty in pruning areas of the tree facing fakultet, Čačak.
inwards of the row and the frequent inability to
use adequate disease control imply the
178
Hrotkó K., Magyar L., Simon G., Klenyán T., 1998. Mitrović M., Blagojević M., Karaklajić-Stajić Ž.,
Effect of rootstocks on growth of plum cultivars in a Rakićević M., 2005. Sistem guste sadnje u
young orchard. Acta Horticulturae, 478: 95-98 savremenoj tehnologiji gajenja šljive. Traktori i
Lučić P., Đurić Gordana, Mićić N., 1996. Voćarstvo I. pogonske mašine, 10 (2): 103-107.
Institut za istraživanja u poljoprivredi Srbija: Nolit- Paunovic G., Milosevic T., Glisic I., 2011.
Partenon. Morphometric traits of newly bred rootstocks suckers
Mićić N., Đurić G., Cvetković M., 2005. Sistemi gajenja in domestic and cherry plum. Acta Scientiarum
i rezidba šljive. Ministarstvo poljoprivrede, Polonorum, Hortorum Cultus, 10 (2): 203-212.
šumarstva i vodoprivrede Republike Srbije, Beograd,
1-60.
179
180
SENSORIAL EVALUATION OF 26 HIGHBUSH BLUEBERRY VARIETIES

IN ROMANIA
Adrian ASĂNICĂ


Abstract
Among many blueberry growers and general consumers perception, often are cross-cutting issues. The quality of the
fruits depends on many factors such as variety, area were fruits are produced (environmental conditions), ripening and
harvest time, producing technology etc. Of course many other influencing factors could intervene during the picking
time till consumption of fruits and affects the fruit appearance, firmness, colour or other internal or external
parameters. Besides all these reasons, the final consumer has particular demands and preferences which stays at the
base of their choices. In 2017, a tasting session with 26 blueberry varieties cultivated in Romania were evaluated by
different categories of consumers. Cultivar tested were: ʻCovilleʼ, ʻChandlerʼ, ʻAzurʼ, ʻDarrowʼ, ʻBluerayʼ, ʻLegacyʼ,
ʻSimultanʼ, ʻBrigittaʼ, ʻAugustaʼ, ʻSpartanʼ, ʻVitalʼ, ʻPembertonʼ, ʻDeliciaʼ, ʻRekaʼ, ʻToroʼ, ʻBerkeleyʼ, ʻDenise blueʼ,
ʻDukeʼ, ʻSafiʼr, ʻLaxʼ, ʻHannah’s choiceʼ, ʻBluecropʼ, ʻPatriotʼ, ʻNelsonʼ, ʻBluettaʼ, ʻPink lemonadeʼ. All these varieties
were evaluated against fruit size, skin colour, firmness, juiciness, taste and flavor. ʻCovilleʼ and ʻChandlerʼ performed
better as general total score. ʻChandlerʼ and the Romanian variety ʻAugustaʼ remarked by large fruits and were
appreciated accordingly. ʻCovilleʼ and ʻBluerayʼ showed good appearance of the skin colour since the most firm fruits
were noticed at ʻLegacyʼ. Most tasty and aromatic fruits were scored for ʻCovilleʼ.
Key words: Vaccinium corymbosum L., organoleptic assessment, consumer preferences.
INTRODUCTION Evaluation of blueberries traits is a

continuously process required by the breeders,
As many other fruits, blueberries are subject of producers and consumers. Many studies were
organoleptic analysis (Polashock J. et al.). conducted in this regard (Itle R.A. and NeSmith
General consumers perception is of great D.S., 2017) trying to emphasize which is the
interest when it comes to retailers and best solution and what varieties are most
influences the fresh fruit market. But, even suitable for a specific regions, countries or
with the latest perfomant lab equipments consumer preference.
(Saftner R. et al., 2008) the consumers Therefore, the main objective of the present
preference is stil a cross-cuting issue. It was study was to investigate the general consumers
demonstrated that major traits of the varieties preference in Romania for the fresh blueberry
that influence the positive perception of fruits fruits since the last 10-15 years, the blueberry
quality are the taste, fruit size, color and flavor consumption increase worldwide very fast.
(Gilbert J.L. et al., 2015). A subsequent goal was also to decelate the
Indeed, the quality of the fruits depends on consumers preference regarding each criteria of
many factors such as area were fruits are assessment. This could be also relevant for the
produced (environmental conditions), ripening breeders when they decide to release new
and harvest time, producing technology etc. varieties on the market.
Many other influencing factors could intervene
during the picking time till consumption of MATERIALS AND METHODS
fruits and affects the fruit appearance, firmness,
colour or other internal or external parameters. In July 17, 2017, a tasting session with
Besides all these reasons, the final consumer different 26 blueberry varieties cultivated in
has particular demands and preferences which Romania (pot crop system), Dambovita county
stays at the base of their choices (Yue C. and were evaluated by different categories of
Wang J., 2017). consumers: students, professors, researchers,
181
farmers and regular people. Also it was a 10 points have been allocated to the fruit
balanced distribution between young, mid age appearance (fruit size plus skin color), 6 points
and elder people as long as male/female to consistence/texture (firmness and juiciness)
participation. and 12 points to taste (taste and flavour). The
The blueberry varieties tested were: ‘Coville’, varieties were reviewed by 25 assessors
‘Chandler’, ‘Azur’, ‘Darrow’, ‘Blueray’, delivering at the end the sheets with the scores
‘Legacy’, ‘Simultan’, ‘Brigitta’, ‘Augusta’, for each analysed item.
‘Spartan’, ‘Vital’, ‘Pemberton’, ‘Delicia’,
‘Reka’, ‘Toro’, ‘Berkeley’, ‘Denise blue’, RESULTS AND DISCUSSIONS
‘Duke’, ‘Safir’, ‘Lax’, ‘Hannah’s choice’,
‘Bluecrop’, ‘Patriot’, ‘Nelson’, ‘Bluetta’, ‘Pink In the first step, all the scores were summed
lemonade’. Seven of them are Romanian and the varieties ranked from the highest value
varieties and the other 19, international to the lowest one.
blueberry varieties (Figure 1). The overall chart highlight ‘Coville’ variety
that gathered 23.18 points out of maximum 28
points (Figure 2) followed closer by ’Chandler’
with 22.43 total points.
Both varieties showed very large fruits and
confirms the appreciation of general consumers
for the higher size of the fruits.
Nevertheless, the appearance of this particular
two varieties was enhanced by the silver pruin
of the skin and the freshness look.
At the bottom of the list, was sitated a novel
blueberry variety ‘Pink lemonade’ with pinkish
and aromatic fruits that did not comply with the
consumers expectation.
Figure 1. The 26 blueberry varieties in the scene of ‘Bluetta’ as a early variety was overripened at
reviewers (17.07.2017)
the time of consumption and therefore has
registered less points for most of the criteria.
All these varieties were evaluated against six It is remarkable the presence of two Romanian
criteria: fruit size (1…5), skin color (1…5), blueberry varieties (‘Azur’ and ‘Simultan’) in
firmness (1…3), juiciness (1…3), taste (1…7) the top 10 most appreciated ones by evaluators.
and flavor (1…5). From the total of 28 points,
Chandler
Pink Lemonade25,00 Augusta (RO)
Bluecrop Azur (RO)
20,71
18,3620,00 22,43 21,82
Nelson - Pemberton
20,21
Hannah's choice18,75 15,00
9,39 20,93 Brigitta
12,50
Lax (RO) 18,82 10,00 21,50 Darrow
5,00
Safir (RO)19,11 20,32 Vital (RO)
0,00
Reka19,86 21,18 Legacy
11,96
Bluetta 23,18Coville (RO)
19,14 19,36
Duke 12,82 Berkeley
Simultan (RO)
21,11 21,29Blueray
19,29
Patriot - 19,71 20,14 Denise blue
Toro 20,64 Delicia (RO)
Spartan
Figure 2. The overall total scores of the tested blueberry varieties

182
In relation with the fruit size, blueberry ‘Augusta’ followed the first variety in the class
varieties ranged between 1.86 at the ‘Pink (Figure 3). Together with ‘Azur’ situated in the
lemonade’ and 4.68 points at ‘Chandler’ out of third position indicate that national varieties
the maximum 5 points foreseen in this criteria. also could be reliable on the international
For only 0.04 points less, the Romanian variety competitive market.
5,00 4,68 4,64 4,54 4,50

4,36 4,32 4,32 4,25 4,21 4,18
4,50 4,11
3,86 3,75
4,00 3,61 3,61 3,61 3,57 3,43
3,25 3,04
3,50 2,89 2,86 2,86 2,82 2,75
3,00
2,50 1,86
2,00
1,50
1,00
0,50
0,00
Figure 3. The ranking list of the blueberry varieties according to the fruit size
In the overall appearance perception, the color is an advantage for the varieties like ‘Coville’,
of the fruits skin is very significant. Thus, a ‘Blueray’ or ‘Safir’ (Figure 4).
proeminent uniform blue of the blueberries skin
5,00 4,54 4,39 4,36

4,39 4,32 4,29 4,21 4,11
4,50 4,04 4,00
4,07 3,93 3,89 3,86 3,82 3,82 3,79
4,00 3,75 3,57 3,54 3,54
3,46 3,43
3,50 3,11
3,00 2,79 2,71
2,50
2,00
1,50
1,00
0,50
0,00
Figure 4. The arch of the blueberry varieties ranked by skin color total scores
The fruits firmness is nowadays, beside the too (Strik B.C. et al., 2017). ‘Reka’ and the
shelf life of the product, more and more desired Romanian variety ‘Delicia’ took an upper
in the fresh consumption. In this respect we position in the ranking list, next by ‘Chandler’
assist at a different rank of the blueberry and ‘Duke’, varieties that are well known for
varieties when it comes to a crispy pulp. In the their firm fruits (Figure 5).
top of the list, ‘Legacy’ confirms the texture
preference of the evaluators and the literature
183
2,82
3,00
2,79 2,79
2,64 2,61 2,61 2,57 2,57 2,57
2,50 2,50 2,46 2,43
2,39 2,39 2,32 2,32
2,50 2,25 2,25 2,21 2,18
2,07
2,00
1,68
1,50
1,50 1,25
1,00 0,82
0,50
0,00
Figure 5. The variability of pulp firmness for the 26 blueberry varieties
The consistence of the fruits is direct so, it were remarked old varieties like ‘Darrow’
influenced by the variety, harvest time, post- or ‘Brigitta’ next by four Romanian blueberry
harvest storage conditions and many other varieties: ‘Vital’, ‘Lax’, ‘Simultan’ and ‘Azur’.
factors that intervene in the process. Therefore, Except few varieties like ‘Toro’, ‘Nelson’,
the values registered by the varieties tested in ‘Patriot’, ‘Bluetta’ and ‘Pink lemonade’, al the
the same day are in a flesh juiciness stage other varieties did not vary too much in the
according to their postharvest evolution. Even scores given by the evaluators (Figure 6).
3,00
2,64
2,61 2,54 2,43 2,39
2,50 2,50 2,46 2,43 2,43 2,39 2,36 2,36 2,36 2,36
2,36 2,21
2,21 2,182,18
2,14
2,00 1,93
1,86
1,50
1,21
1,07
1,00 1,04
0,82
0,50
0,00
Figure 6. The juiciness spectrum of the 26 blueberry varieties
Taste is imediately after appearance the most the variety when the client wants to buy again.
important trait of the fruit assesed by the In this regard, a special attention was payed by
consumer. the breeders trying to match the consumer
It is definetely one of the criteria and reason for preference. The results obtained in our tasting
the consumer to return and taste again. It session restore the performance of an old
represents the path to remember the name of blueberry variety, famous for taste and medium
184
to large fruits, namely ‘Coville’. It convinced in points). Both are Romanian varieties less
a significant and evident way by gathering no popular that other foreign varieties. ‘Augusta’
less than 5.36 points (Figure 7) out of the performed better due to its special and
maximum 7 followed at some distance by particular taste and flavor.
‘Augusta’ and ‘Azur’ with the same score (4.68
1,64
Pink Lemonade 1,68
1,86
Nelson - 2,00
3,61
Bluecrop 3,75
3,86
Berkeley 3,89
3,93
Safir (RO) 3,93
4,00
Denise blue 4,07
4,07
Pemberton 4,11
4,11
Brigitta 4,21
4,32
Simultan (RO) 4,32
4,36
Blueray 4,39
4,43
Azur (RO) 4,54
4,64
Augusta (RO) 4,68
4,68
Coville (RO) 5,36
0,00 1,00 2,00 3,00 4,00 5,00 6,00
Figure 7. The ranking of all 26 bluebery varieties according to the consumers taste preference
The flavor of the fruits (Figure 8) completes the it was expected, the first two varieties ‘Coville’
taste and inprint the consumer memory. As and ‘Augusta’ were assigned as most flavored
many regular and unadvised consumer, the varieties recording the highest scores.
taste results are similar with the flavor one. As
Coville (RO)
1,36
1,501,46 4,21 Augusta (RO)
1,68 3,89
3,07 Chandler
3,11 3,86
Simultan (RO)
3,18 3,86 Spartan
3,25 Azur (RO)

3,75
Darrow
3,32
3,68 Hannah's choice
3,43 Brigitta
3,61 Blueray
3,43
Denise blue
3,57
3,43 Lax (RO)
3,54
3,46 Safir (RO)
3,50
3,46 3,50 Toro
3,46
Reka
Figure 8. Consumers preference range for the blueberry flavor assesed in the tasting session
185
It is again remarkable the fact that ‘Coville’ REFERENCES
detached in a significant way from the second
position also for this item analysed. Another Gilbert J.L., Guthart M.J., Gezan S.A., Pisaroglo de
Romanian blueberry variety (‘Simultan’) Carvalho M., Schwieterman M.L., Colquhoun T.A.,
Bartoshuk L.M., Sims C.A., Clark D.G., Olmstead
remains in the top preference of the consumers J.W., 2015. Identifying Breeding Priorities for
for the flavor, considered one of this variety Blueberry Flavor Using Biochemical, Sensory, and
major advantage. Genotype by Environment Analyses. PLoS ONE
10(9): e0138494.
CONCLUSIONS https://doi.org/10.1371/journal.pone.0138494
Itle R.A., NeSmith D.S., 2017. Evaluation of fruit quality
traits in southern highbush and rabbiteye blueberries.
‘Coville’ and ‘Chandler’ varieties performed Acta Hortic. 1180, 393-400
better as overall total score and taste in the DOI: 10.17660/ActaHortic.2017.1180.54
consumers preference range. Polashock J., Saftner R., Ehlenfeldt M. Organoleptic
‘Chandler’ and the Romanian variety ‘Augusta’ analysis of blueberry cultivars, Rutgers New Jersey
Agricultural Experiment Station, 152-153.
were remarked for their large fruits. Saftner R., Polashock J., Ehlenfeldt M., Vinyard B.,
‘Coville’ and ‘Blueray’ showed good appea- 2008. Instrumental and sensory quality characteristics
rance of the skin color. of blueberry fruit from twelve cultivars. Post-harvest
The firmest fruits were noticed at ‘Legacy’ and Biology and Technology, Volume 49, Issue 1, 19-26.
‘Reka’. Strik B.C., Vance A.J., Finn C.E., 2017. Northern
Highbush Blueberry Cultivars Differed in Yield and
Most tasty and aromatic fruits were ones from Fruit Quality in Two Organic Production Systems
‘Coville’ and again ‘Augusta’. from Planting to Maturity, Hort. Science 52, 844-851
High aromatic fruits were positive evaluated at Yue C., Wang J., 2017. Consumer preferences for fresh
‘Coville’, ‘Augusta’ and ‘Chandler’. blueberry attributes. Acta Hortic. 1180, 1-8 DOI:
Evaluation of the blueberry varieties from the 10.17660/ActaHortic.2017.1180.1
consumers preference point of view has to be
understood as a dinamic process and must be
periodically repeated.
ACKNOWLEDGEMENTS
This work was supported by a grant of the

Romanian National Authority for Scientific
Research and Innovation, CNCS-UEFISCDI,
project number PN-II-RU-TE-2014-4-0749.
186
EVALUATION OF THE CONSUMER PREFERENCE

FOR SWEET CHERRY FRUITS AT THE “SWEET CHERRY FEST”
IN ISTRIŢA - BUZĂU
Adrian ASĂNICĂ, Florin STĂNICĂ, Valerica TUDOR,

Alexandru IACOB, Violeta ZOLOTOI, Livia PEROJUC

Abstract
Since 2013, the Univeristy of Agronomic Sciences and Veterinary Medicine of Bucharest yearly organizes at the Istriţa
Nursery and Didactic-Experimental Farm the “Sweet Cherry Fest” in the first decade of June. The event is part of the
“Agriculture for Life, Life for Agriculture” International Conference programme and consist in a large sweet cherry
exhibition and tasting of fresh sweet cherry fruits gathered from own trial orchards or from comercial sweet cherry
plantations within the country. Depending on the environmental conditions and weather conditions of each year, the
samples of cherries for tasting session varies a lot but comprise as many as possible varieties, most of them from early
to medium harvest time. In 2017, the tasting panel evaluated 24 samples coming from 17 varieties. These were as
follows: ʻEarly redʼ, ʻBurlatʼ, ʻCelesteʼ, ʻKordiaʼ, ʻFirm redʼ, ʻGrace Starʼ, ʻNew Starʼ, ʻEarly Bigiʼ, ʻGiant redʼ,
ʻSummitʼ, ʻFerroviaʼ, ʻUlsterʼ, ʻVanʼ, ʻRubinʼ, ʻKatalinʼ, ʻSkeenaʼ and ʻLapinsʼ. Best score was recorded by ʻEarly redʼ
followed by ʻNew Starʼ and ʻGiant redʼ. The fruit size and appearance of the ʻEarly redʼ and ʻGiant redʼ varieties were
rewarded by tasting panel with high appreciation scores. The taste was also remarked by the consumers for ʻEarly redʼ
and also the aroma for ʻNew Starʼ.
Key words: variety, organoleptic assesment, taste, fruit appearance, consumer preference.
INTRODUCTION analyses taken by panelists versus common

consumers support retailers, farmers or
Since 2013, the Univeristy of Agronomic breeders to figure out the key drivers for sweet
Sciences and Veterinary Medicine of Bucharest cherry market need (Revell J., 2008).
yearly organizes at the Istrita Nursery and Different compounds of the fruits are identified
Didactic-Experimental Farm the “Sweet Cherry (Legua P. et al., 2017) and associated with the
Fest” in the first decade of June. consumers preference (Vavoura M.V. et al.,
The event is part of the “Agriculture for Life, 2015; Serradilla M.J. et al., 2012).
Life for Agriculture” International Conference But characterization of sweet cherry cultivars
programme and consist in a large sweet cherry only by lab instruments is unpaired without
exhibition and tasting of fresh sweet cherry tasting evaluation session.
fruits gathered from own trial orchards or from
commercial sweet cherry plantations within the MATERIALS AND METHODS
country.
Depending on the environmental conditions The exhibition and tasting session within the 4th
and weather conditions of each year, the edition of the “Sweet Cherry Fest” took place
samples of cherries for tasting session varies a in Istrita Nursery and Farm in 10th of June,
lot but comprise as many as possible varieties, 2017.
most of them from early to medium harvest The tasting panel consist of 44 persons which
time. assessed 24 samples represented by 17 varieties
Besides many researches conducted in this field (Figure 1). The gender balance was good, in the
(Amidei R. et al., 2017; Ross C.F. et al., 2010; session participating 25 male and 19 female
Chauvin M.A. et al, 2009), the sensorial above 18 years.
187
Sweet cherry varieties introduced in the contest The external traits gather 10 points, the internal
were as follows: ‘Early red’, ‘Burlat’, ‘Celeste’, characters 9 points and the taste 7 points
‘Kordia’, ‘Firm red’, ‘Grace Star’, ‘New Star’, maximum plus 4 points from the flavor.
‘Early Bigi’, ‘Giant red’, ‘Summit’, ‘Ferrovia’, Total score is 30 points.
‘Ulster’, ‘Van’, ‘Rubin’, ‘Katalin’, ‘Skeena’
and ‘Lapins’.
Figure 1. Participants during the tasting session -

10.06.2017 at Istrita, Buzau
The samples provenience were mainly from the

University of Agronomic Sciences and
Veterinary Medicine Bucharest. Figure 2. Tasting sheet used by the participants in the
The host - Istrita Nursery and Fruit growing sweet cherry quality assessment
Farm provided nine varieties and the Faculty of
Horticulture Bucharest came with 10 sweet RESULTS AND DISCUSSIONS
cherry varieties from the didactic and experi-
mental field situated in the Bucharest campus. All the samples accepted in the contest were
Two private companies: “Livada fermecata” scored by the evaluators and centralized after
Agroclaas SRL (Haret, Vrancea) and Multifruct collecting the sheets.
SRL (Chiselet, Calarasi) introduced in the Summarizing the scores from each item we
tasting session another five varieties. obtained the total scores that ranked better the
From the Research and Development Fruit ‘Early red’ variety from the Istrita Nursery
Growing Station Iasi, the varieties and hybrids and Farm (27.25 points out of max 30 points)
were presented only in the exhibition. (Figure 3).
The samples were encoded and each participant The second best variety was ‘New Star’ from
move from one variety to another marking in the Istrita Nursery and Farm with 26.44 points
the tasting sheet their appreciation (Figure 2). followed by ‘Giant red’ from the same
In total, eight items were analyzed for the sweet institution (25.51 p).
cherry assessment.
The fruit size, appearance, skin color, firmness,
juiciness, juice color, taste and flavor.
188
Figure 3. The distribution of total scores among all sweet cherry samples participating in the tasting evaluation session (2017)
Considering the size of the fruit, the highest In terms of appearance of the fruits, the most
scores were registered by ‘Early red’ (Figure appreciated variety was ‘Burlat’ from the
4), ‘Firm red’ and ‘Giant red’ from the Istrita “Livada fermecata”, Haret, Vrancea which gain
Farm and Nursery Station. Bigger fruits were an average of 2.94 points out of maximum 3.0
also remarked at ‘Burlat’ from “Livada points. In the second position was notced
fermecata” and ‘Grace star’ from Multifruct ‘Early red’ from the Istrita Farm and Nursery
SRL. and in the third level ‘Celeste’ from the same
institution.
The highest appreciation for the skin color was
highlighted by the same ‘Early red’ from Istrita
followed closely by ‘Burlat’ from “Livada
fermecata” (Figure 4).
Surprisingly, ‘Ferrovia’ from the Multifruct
SRL impress the tasters with a very firm fruits
(2.81 p out of 3.0 p). The second place was
ocupied by ‘Firm red’ from Istrita, the name of
the variety is in this way reconfirmed.
‘Celeste’ from Istrita was the variety with
higher content of juice in the perception of
assessors together with ‘Early Bigi’ from
Multifruct SRL.
The color of juice emphasized ‘Burlat’ from
“Livada fermecata” and ʻEarly red’ from Istrita
followed by ‘Kordiaʼ and ‘Celeste’ from Istrita
too.
The most important feature of the sweet cherry
fruit is the taste. As we expected, ‘Early red’
from Istrita was very much appreciated, with an
Figure 4. The appearance and fruit size average score of 5.98 p out of 7.0 p. ‘Burlat’
of ‘Early redʼ variety even that is an old variety earn the respect
189
of the evaluators which scored with 5.42 p. The biggest fruit size was remarked at ‘Early
(Figure 5). Red’, followed by ‘Firm red’ and ‘Giant red’
varieties from Istrita Nursery and Farm.
In terms of fruit appearance and skin color,
‘Burlat’ from “Livada fermecata” and ‘Early
red’ from Istrita were rewarded by the
evaluators with higher scores.
The firmest pulp had ‘Ferrovia’ from
Multifruct SRL and ‘Firm red’ from Istrita.
The taste and aroma of ‘Early red’, ‘Burlat’ and
‘Kordia’ was remarked and highly appreciated
by the consumers.
REFERENCES
Amidei R., Castellari L., Missere D., Grandi Mi., Lugli

S., 2017. Fruit sensory test of new sweet cherry
cultivars. Acta Hortic. 1161, 593-598 DOI:
10.17660/ActaHortic.2017.1161.94
Chauvin M.A., Whiting M., Ross C.F., 2009. The
Influence of Harvest Time on Sensory Properties and
Consumer Acceptance of Sweet Cherries, Hort
Technology October-December, vol. 19 no. 4 748-
754.
Figure 5. Variety ‘Burlat’ from “Livada fermecata”, Legua P., Domenech A., Martínez J., Sánchez-Rodríguez
Vrancea L., Hernández F., Carbonell-Barrachina A.,
Melgarejo P., 2017. Bioactive and Volatile
‘Kordia’ and ‘Celeste’ from Istrita also were Compounds in Sweet Cherry Cultivars. Journal of
Food and Nutrition Research, 5 (11), 844-851.
remarked from this point of view. The most Revell J., 2008. Sensory profile & consumer
disliked varieties were ‘Lapins’, ‘Katalin’ and acceptability of sweet cherries. Master thesis
‘Ferrovia’. University of Nottingham.
A strong correlation between taste and aroma Ross C.F., Chauvin M.A., Whiting M., 2010.
was found, most of the evaluators ranking Assignation of sweet cherry selections to 3 taste
groupings based on perceived sweetness and
higher the same varieties of sweet cherry. sourness, J Food Sci. Jan-Feb;75 (1): S48-54. doi:
Therefore, ‘Early red’ and ‘Burlat’ plus 10.1111/j.1750-3841.2009.01410.x.
‘Kordia’ and ‘Celeste’ were considered the Serradilla M.J., Martín A., Ruiz-Moyano S., Hernández
most aromatic fruits between the samples. A., López-Corrales M., de Guía Córdoba M., 2012.
Physicochemical and sensorial characterisation of
four sweet cherry cultivars grown in Jerte Valley
CONCLUSIONS (Spain), Food Chemistry, Volume 133, Issue 4, 1551-
1559.
Best total score was recorded by ‘Early Red’ Vavoura M.V., Badeka A.V., Kontakos S., Kontominas
from Istrita Nursery and Farm followed by M.G., 2015. Characterization of Four Popular Sweet
‘New Star’ and ‘Giant red’ from the same Cherry Cultivars Grown in Greece by Volatile
Compound and Physicochemical Data Analysis and
institution. Sensory Evaluation. Molecules, 20, 1922-1940.
190
THE BEHAVIOR OF GALA, JONAGOLD, GOLDEN DELICIOUS AND

GRANNY SMITH APPLE VARIETIES IN ORGANIC FARMING SYSTEM
Andreea BUJDEI1, Roxana CICEOI2, Florin STĂNICĂ1

1
University of Agronomic Sciences and Veterinary Medicine of Bucharest, 59 Mărăști Blvd,
011464, District 1, Bucharest, Romania
2
University of Agronomic Sciences and Veterinary Medicine of Bucharest, Research Center for
Studies of Food Quality and Agricultural Products, 59 Mărăști Blvd, 011464, Bucharest, Romania

Abstract
In the present study, the behaviour of the ʻGalaʼ, ʻJonagoldʼ, ʻGolden deliciousʼ and ʻGranny Smithʼ apple varieties,
grown in an organic orchard in Arad County, Romania is presented. The intensive orchard was planted in 2010, on
3.275 ha with apple trees, grafted on M9 rootstock, using 2.6 m x 0.8 m planting distances and 3.5 m high concrete
poles with 5 lines of wires, as trellis system. The total yield and productivity of the apple varieties are presented and
compared with the initial data estimated in the business plan of the structural funds' project. 2017 was the first year in
which the total harvested production surpassed the estimation from the business plan, seven years after planting, but
the overall real total production was still 68.1 tones, lower than previsioned. Fruit quality was reduced due to scab
attack symptoms, sunburns, cracking and low calibre. Even so, by using intelligent marketing strategies as “buy
locally” and “pick it yourself” besides the organic distribution chains, the economic losses were diminished.
Considering our results, we do not recommend the cultivation of high disease sensitive apple varieties in organic
orchards in areas with high infection pressure and without an efficient phytosanitary protection. The cultivation of scab
resistant varieties and a proper orchard management could be the solution for the development of the organic apple
production.
Key words: apple production, high density, productivity, fruit quality.
INTRODUCTION farming, including advantages offered by

different EU structural funds measures for
As eating healthy and life quality becomes agriculture for those activating in the organic
more and more important, the market shifts its sector. For the period 2007-2013 the
focus and the business owners reorient procentage of the European Union subsidy
themselves towards the organic products and received by Romania was 52.02% in average
services that satisfy the clients needs. EU (Marinas et Prioteasa, 2016), altghough some
makes serious efforts to fulfill their citizen afirm. For the Measure 1.2.1 (Agriculture), in
expectations and promote and support the Octomber 2016, in Romania, out of 8738
organic farming developpment. The organic projects proposals, 3849 were aproved and
farming in Romania is still at its beginning, the 2789 finalised, with a total of 662,274,906 euro
dynamic of organic production indicators payments.
showing different fluctuations, except the Apple is the most important fruit species in
organic fruit and vineyard areas, where the Romania, our country having the second
trend was constantly ascending (MADR, 2018). highest area cultivated with the apple in Europe
This ascending situation was also sustained by (after Poland), representing a share of 10.2% of
the EU structural funds given for agriculture the total EU-28 production area and only a
and rural development. Although the total share of 3.6% of the total EU-28 harvested
organic area cultivated in Romania decreased production (EUROSTAT, 2016).
with 21.5% in the period 2012-2016 Organic fruit growing has increasingly gain
(EUROSTAT, 2017), the organic orchard and interest both from Romanian and European
vineyard area tripled in the same period farmers or investors, althgough uncertified
(Burghelea et al., 2016). Romania makes organic food (as farmers’ market, self-
serious efforts to support and promote organic production etc.) is considered by consumers to
191
be more “organic” than certified organic food ha) in four days. On the 90 m x 360 m plot 3
(Petrescu et al., 2016). years old plants of ʻGalaʼ, ʻJonagoldʼ (ʻRed
The company Fruit4you, Belgian investors, Princeʼ) ʻGolden deliciousʼ and ʻGranny Smithʼ
which owns the 3,275 ha of organic apples in varieties, grafted on M9 rootstock, were
Horia village, Vladimirescu commune, Arad planted.
county, received more than 350,000 euro for The investors preferred to plant at 2.6 m x 0.8
the implementation of their organic apple m, for a „fruit wall shape” high density
orchard. orchard, with 4,000 trees/ha, having 3.5 m high
In this paper, we present the economic results concrete poles and 5 lines of wires. The poles
of the organic apple orchard in Arad County, in were distributed every 10 m, for 360 m, with a
terms of production, productivity, income in space in the middle of the orchard, where the
the 2011-2017 period and we compare these tractor can make the “U” turn. The varieties
results with the previsions made in the were planted by groups of four rows, starting
structural funds business plan. and the scheme repeated for 27 lines. The inter
rows were mowed mechanically and the row
MATERIALS AND METHODS was cleaned by hand.
Fertirrigation was insured by drip irrigation
The data used for this study were gathered from lines placed directly at the ground level,
the apple orchard belonging to company working on 4 sectors, every day, with 2 hours
Fruit4you, lat: 46.1986, long: 21.4285 (Figure of watering per sector.
1). As the investors were from Belgium, the The hail and pest protection was insured by a
same technology as in Belgium was used for hail protection net that covers the top and the
the apples plantation. The soil was prepared in four laterals of the orchard (Figure 2).
the spring and the planting was done with a
planting machine.
Figure 1. The Fruit4you orchard
The trees were imported from a Belgian

nursery, transported in a temperature-controlled
cargo and kept at 25-28°C (the normal
temperature environment), after arrival, with Figure 2. The trellis and the hailnet system
the roots in water for 24 hours to get hydrated.
The planting was done in June-July 2011, No tree pruning was done in the first year. In
because of the delays in trees delivery. The late the second year the trellis system was put in
planting period caused a premature period of place, the first three wires, followed by the next
dormancy, which made the young trees fourth and fiftth wires in the third year.
apparently looking as they were died in the The canopy forming prunings were performed
autumn. The delay in plants delivery also after the Belgian method, during the flowering
forced the investor to build a buffer for the time, which also implies slow shoot growths of
irrigation system that collects the water from around 30 cm/year.
the well and reduce the water temperature As the orchard is included in the organic
difference before the irrigation. The planting farming system, the insects were controled with
machine, with an potential efficiency of 4,800 pheromone disruptors and the hail nets, that
trees/day manage to plant the orchard (3,275 were kept closed from May 1st to September
192
30.800 units/ha of Isomate CLR mating A similar high yield was reported by Sumedrea
disruptors (active substance codlemone), were et al. in 2016, a production of 44.40 t/ha for
used each year, being placed at height of the 3rd ʻGolden deliciousʼ Clone B. As in the period
wire (~ at 1.8 m), one every 3 trees, one the 2012-2015, the productions were lower than
edges and one every 5 or 8 trees in the centre of expected, the producer tried in one year, on one
the orchard. Every year the dispensers were put row, a production without thinning.
in place at the beginning of May (Bujdei et al., The result was promising, with 35% higher
2016). The insects attack was estimated under production on that row, but in the following
1%, in the flowering period only, when the net year no production was obtained, as no
is still closed, for a better pollination. flowering buds were differentiate.
After the net was closed, no insect issues were
observed in all 6 years of cultivation. Mineral Table 2. The estimated and realized productions
of each apple variety in the 2011-2017 period
oil was sprayed in spring 2012 and treatments
with cooper (Bouille Bordelaise WDG) were Year Golden Gala Jonagold Granny Total yield
(t) (t) (t) Smith (t) (t)
applied during the seasons. 2011 1.00 1.00 1.00 0.30 3.30
The fruits were picked at maturity and kept in 2012 4.50 5.40 5.10 0.90 15.90
cold storage, at 4°C. 2013 3.00 9.00 11.40 - 23.40
2014 8.40 8.40 10.50 - 27.30
RESULTS AND DISCUSSIONS 2015 13.50 16.50 12.60 - 42.60
2016 15.00 17.40 9.00 - 41.40
2017 48.60 53.00 57.40 - 159.00
Our paper presents the real experience of one Total
applicant to 1.2.1 Rural Development Measure, yield/ 94.00 110.70 107.00 312.90
focusing on the real challenges that one variety
investor met within an organic apple orchard. Average
Yield, productivity and economic efficiency yield 14.35 16.90 16.34
/ha
The average fruit production/ha was estimated
at 18.58 t/ha in the business plan and the
realized production was of 15.91 t/ha, with ʻGalaʼ variety was the most productive one,
14.35% less (Table 1). The differences were with a productivity of 16.90 t/ha, calculated for
much higher in the first five years, only 50% of the 2011-2017 period (Table 2).
what was estimated was realised until 2016. The ʻGranny Smithʼ variety was replaced after
The year 2017 was a very good year for apple the first 2 years, due to its scab sensitivity and
growing in the region with an average annual low production. Because of scab attack, the
production of 48.55 t/ha, which represented a general aspect of the trees and apples was
record. unsatisfactory, which forced the producer to
sell the ʻGranny Smithʼ apple at a reduced price
Table 1. The estimated and the realized (50% lower than expected). Although this
apple production in the 2011-2017 period variety should produce big apples, no fruit was
bigger than 65+ mm (Figure 3).
Difference
Difference
Real yield
Estimated
yield
Year
(t)
(t)
(t)
2011 3.03
2012 30.00 -14,10 -47.00
2013 55.00 23.40 -31.60 -57.45
2014 70.00 27.30 -42.70 -61.00
2015 70.00 42.60 -27.40 -39.14
2016 70.00 41.40 -28.60 -40.86
2017 70.00 159.00 89.00 127.14
Total yield 365.00 312.63 -52.37 -14.35
Total annual
yield 60.83 52.11 -8.73 -14.35
Annual
yield/ha 18.58 15.91 -2.67 -14.35 Figure 3. Small fruits produced by ʻGranny Smithʼ
193
ʻJonagoldʼ had a lower productivity when Crop issues
compared with ʻGalaʼ, but the apples were in The most frequent crop issues that the producer
majority of 1st category (80+ calibre) (Figure 4). faced were sunburns, cracking and scab
ʻGolden deliciousʼ had in average 65% scab (figures 5, 6 and 7) and the most susceptible
free apples, with fruits varying in size between variety was ʻGalaʼ.
65 and 80+mm. ʻGalaʼ was more affected by
scab, with around 45% scab free apples and the
calibre between 60 and 70 mm.
Figure 5. Sun burns and cracking on ʻGalaʼ variety
Figure 4. The calibre measurments for ʻJonagoldʼ apples
Regarding the income, in the period 2012-2016

all our estimations were unrealistic, as the real
income between 39 and 61% of the estimated
one (Table 3).
Table 3. The estimated and actual orchard incomes in the Figure 6. Scab symptoms on ʻGalaʼ variety leaves
2011-2017 period
Year/ to Estimated Real Difference Difference Regarding scab, more than 45% of the ʻGalaʼ,
income income (euro) (%) more than 35% of ʻGoldenʼ and more then 15%
(euro) (euro)
of ʻJonagoldʼ fruits had the disease marks, a
2011
2012 17,523 4,437 -13,086 -74.68
fact wich led to a loss due to the lower selling
2013 32,126 5,986 -26,140 -81.37 price.
2014 40,887 13,029 -27,858 -68.13
2015 40,887 17,427 -23,460 -57.38 Selling strategies
2016 40,887 14,113 -26,774 -65.48
2017 40,887 42,400 +1,513 +3.70
The producer mainly used two selling
strategies: “buy locally” and “pick it yourself”.
The average price of apples was 2.5 lei/kg (55
euro cent/kg) and the producer was able to
maintain this price only because he has built
through the structural funds a cold storage, that
allowed him to keep and sell the fruits later
than all the other producers in the area and also
to deliver in high quantities at once, loading a Figure 7. The “buy locally” and “pick it yourself”
full truck. Still, the price was not as expected. announcements on Fruit4you Facebook page
Another issue was the lower production
obtained in comparison with the estimated one. The main retailer client was Real Hyper Market
Beside the stress, this fact leads to the loss of Arad, but a part of the fruits also were bought
important contracts and direct income losses. by the Belgian fruits and vegetables dealer,
Fresh Fruit Service BvB.
194
CONCLUSIONS ACKNOWLEDGEMENTS
The estimations in the business plan when the This research work was carried out with the
producer applied to structural funds were more support of SC Fruit For You SRL.
optimistic than the results obtained in reality,
especially regarding the income and this had a REFERENCES
negative impact on the business.
The highly scab sensitive varieties: ʻGalaʼ, Bujdei A., Ciceoi R., Mardare E., Stănică F., 2016.
ʻJonagoldʼ (ʻRed Princeʼ) ʻGolden deliciousʼ Methods of pests prevention and control applied in
two organic apple orchards, Journal of Horticulture,
and ʻGranny Smithʼ faced serious problems Forestry and Biotechnology, 20 (4), 28-32.
under a high infection pressure and the lack of Burghelea C., Uzlău C., Ene C. M, Topor D., 2016.
good protection products. Cooper sulphate Prospects for organic farming in Romania, Scientific
couldn’t offer an effective protection against Papers Series Management, Economic Engineering in
scab and the percentage of affected fruits was Agriculture and Rural Development, 16: 73-78.
EUROSTAT, 2016. The fruit and vegetable sector in the
extremely high, from 15-45%. ʻGranny Smithʼ EU - a statistical overview, http://ec.europa.eu/
was eliminated for this reason, after two years eurostat/statistics-explained/index.php?title=
of cropping. Insects attack on fruits was kept The_fruit_and_vegetable_sector_in_the_EU_-
less than 1% by using winter oil treatment, _a_statistical_overview
matting disruption and net closed orchard. The EUROSTAT, 2017. Organic farming statistics,
http://ec.europa.eu/eurostat/statistics-
level of fruit production and its poor quality explained/index.php/Organic_farming_statistics
affected seriously the orchard incomes, the Marinas L.E., Prioteasa E., 2016. Spotlight on Factors
results being with 57.38-81.37% lower than the Influencing the Absorption Rate of EU Funds in
estimated ones. Only in 2017, the income Romania, Journal of Eastern Europe Research in
overpassed the estimation, due to an Business & Economics, 2016: 1-12, Article ID
500580, DOI: 10.5171/2016.500580
exceptional production. MADR, 2018. Dinamica operatorilor si a suprafetelor in
Taking in consideration our experience, we agricultura ecologica http://www.madr.ro/agricultura-
recommend to other farmers: ecologica/dinamica-operatorilor-si-a-suprafetelor-in-
- to set up of the irrigation drip line at 60-80 cm agricultura-ecologica.html
height instead of laying directly on the soil - no Organic Food Market Development in Romania,
Management Strategies Journal, Constantin
rats eating the tube and no need to massive Brancoveanu University, vol. 32 (2), pages 4-14.
replacement due to destruction during weeding; Romania Insider, 2016. Absorption of EU funds from
- to use higher planting distances between 2007-2013 period reaches 81.73% in Romania,
rows, to at least 3.2 m; https://www.romania-insider.com/absorption-eu-
- to use mechanical equipments for weed funds-2007-2013-period-reaches-81-73-romania/
Petrescu D.C., Petrescu-Mag R.M., Burny P., Azadi H.
control on the fruit tree row; 2016: A New Wave in Romania: Organic Food.
- to associate in growers groups, in order to fill Consumers’Motivations, Perceptions and Habits,
the eventual gaps that some may have in some Agroecology and Sustainable Food Systems,
years, reduce the cost of cold storage and not at DOI:10.1080/21683565.2016.1243602
last, to be able to negotiate with the buyer for Sumedrea D., Chitu E., Chitu V., Sumedrea M., Marin
F., Tanasescu N., 2012. Fruit growing technologies to
fair selling prices. mitigate the negative impact of climatic changes from
The general conclusion is to avoid the the southern part of Romania, Scientific Papers,
cultivation of scab sensitive varieties in organic Series B, Horticulture, Vol. LVI: 185-192.
apple orchards and to replace them with scab
resistant ones. A proper orchard management is
needed to control the phytosanitary issues, to
ensure a proper fertilization and a rational tree
growth and fruit bearing.
195
196
SOME RESULTS ON NUTRITIONAL PROPERTIES OF ORGANIC ROSE

PETALS AND RELATED PROCESSED PRODUCTS
Ana Cornelia BUTCARU, Florin STĂNICĂ, Aurora DOBRIN

Research Centre for Studies of Food and Agricultural Products Quality,
59 Mărăşti Blvd, District 1, Bucharest, Romania,
Abstract
The actual paper presents the first results on the nutraceutical substances found in the rose petals of different varieties,
grown organically in different experimental variants. The petals were collected from each variant and analysed in fresh
and dehydrated status. Rose petal jams produced with different ingredients (ginger, lemon, seabuckthorn) were also
analysed. Total carotenoids content in fresh petals for all ʻCrown Princess Margaretaʼ variants was influenced by the
mulch and ameliorative variants. Dried petals preserved the carotenoids content while in the rose petal jam was very
low. Total anthocyanin content in fresh petals of ʻFalstaffʼ variants were in average at 33.06 mg/100 g. Wool was
beneficial, the maximum values on anthocyanin being registered in these variants. Water content was similar in all
variants with an average of 84.14%. Total soluble solid (TSS) content varied with the mulch variants (wood chips and
wool) and also with the plant ameliorative species. The maximum value obtained was 11.10%. Both rose varieties
analyzed have close TSS values.
Key words: organic horticulture, carotenoids, anthocyanins, rose petal jam
INTRODUCTION ʻFalstaffʼ, created in 1999, is a large and full

petalled flowers with a dark crimson colouring.
From ancient times, roses have been used in The growth is strong and can reach 2.0 - 2.5 m
medical treatments and as food ingredients. high as a climber.
Still now, rose petals and fruits are used in ʻBrother Cadfaelʼ, created in 1986, has a large,
different combinations in pharmaceutical and globular with a clear pink flowers. It has a
food products, being highly appreciated for particulary strong and rich old rose fragance
their bioactive substances: etheric oils, tannins, and can reach 2.5 - 3.0 m high.
carotenoids, anthocyanin, organic acids, ʻCrown Princess Margaretaʼ, created in 1999,
vitamins, minerals etc. (Vasilca Mozăceni, has a strong and fruity fragance like a Tea Rose
2002; Milică et al., 2010; Lambraki, 2001.) type. It grows to 2.5 - 3.0 m high, usually
Rose petals, like the rose fruits, are an import bearing clusters of large apricot colour flowers.
source of antioxidants (Barros et al., 2011). (Austin, 2012; Wagner, 2010).
The actual paper presents the first results on the Before and after establishing the organic rose
nutraceutical substances found in the rose culture, three ameliorative species: Sinapis alba
petals of different varieties, grown organically L., Phacelia tanacetifolia L. and Tagetes patula
in different experimental variants. L. were use to increase the soil biologic activity
and for soil disinfection and disinsection.
MATERIALS AND METHODS All the three ameliorative species were sown in
the spring, by combining the three species in 7
The organic culture was established in 2015 at variants: V1 Sinapis, V2 Sinapis + Phacelia,
the Didactic Experimental Field of the V3 Phacelia, V4 Sinapis + Tagetes, V5 Sinapis
University of Agronomic Sciences and + Tagetes + Phacelia, V6 Tagetes + Phacelia,
Veterinary Medicine of Bucharest, using three V7 Tagetes and a control parcel V8, was kept
climbing David Austin rose varieties: as black field, without sowing.
ʻFalstaffʼ, ʻBrother Cadfaelʼ and ʻCrown They were trimmed and incorporated into the
Princess Margaretaʼ. soil after the flowering period.
197
Starting with 2016, on the rose rows, the soil (2014), Ticha (2015), Bezdadea Cătuneanu et
was mulched with saw dust and wool for each al. (2017).
initial variant (Vn): Vn.1. wood chips and Total anthocyanins content was studied to
Vn.2. wool, while the control Vn.3., was ʻFalstaffʼ variety and was measured with
represented by unmulched soil. Both mulched Specord 210 Plus spectrophotometer at λ = 540
rows had the same 1 m width with the specific nm (Bărăscu et al., 2016; Bezdadea Cătuneanu
material. et al., 2017), after an adapted method. The
For each experimental variant, the same extracts were filtered under vacuum and
technology was applied using manure completed up to 50 ml volume. The results
fertilizing at planting and other organic were calculated using the formula: Total
products in the following years. Plant anthocyanins = DO540 x F, where DO540 is
protection was ensured with copper, sodium absorbance at wavelength λ = 540 nm and
bicarbonate, sulfur, pepper, garlic and propolis factor F = 11.16. The total anthocyanins
tincture. Different biostimulators and caw milk content was expressed in mg/100 g in fresh
were used to increase the plant immunity weight.
system. Total carotenoids were studied to ʻCrown
The petals were collected from each variant Princess Margaretaʼ variety. The identification
begining with May, in the morning (08.00 - and quantification of total carotenoids content
10.00), being collected initially in plastic bags was made after an adapted method after
and transported in cold rooms. Lichtenthaler and Wellburn (1983) and Arnon
A part of the rose petals were dehydrated with (1949).
an Escalibur dehidrator, 3 hours at 40°C. The The analyses were conducted in the Research
dehydrated petals were kept in paper bags at Center for Studies of Food and Agricultural
room temperature. Products Quality, University of Agronomic
From fresh petals were produced more variants Studies and Veterinary Medicine of Bucharest.
of rose petal jam (D1 - ʻCrown Princess
Margaretaʼ, D2 - ʻFalstaffʼ, D3 - ʻBrother RESULTS AND DISCUSSIONS
Cadfaellʼ) with different ingredients: lemon
(Dn.1), ginger (Dn.2), seabuckthorn (Dn.3). For the ʻCrown Princess Margaretaʼ variety
The receipe for every variant consisted in a were analysed total soluble solids, dry matter
base formula (sugar siroup: 1.5 kg sugar at 1.0 l and total carotenoids content in fresh petals,
water with 0,5 kg fresh rose petals) with dry petals and jam comparing the variants of
ingredients added in the final stage ( 0.3 ml ameliorative species and mulch used (Table 1).
lemon juice, 1.2 ml seabuckturn or 50 g.
ginger) Table 1. Influence of variants on evolution of total
This study presents the first results on some soluble solids and dry matter on fresh petals of ʻCrown
Princess Margaretaʼ variety
nutraceutical substances found in rose petals
and rose jams, as total carotenoids and total
anthocyanins, influenced by the three varieties Dry matter Water Total Soluble
Variant content content (U) Solids (TSS)
and the applied organic growing technologies. (D.M. %) (%) (% Brix)
The content in total soluble solids and dry
matter is also reported. V1.1 19.33 80.67 8.90
Total soluble solids were determined from rose V2.1 16.38 83.62 8.50
petal juice (Yoon, 2005; Saei, 2011; Mureşan, V3.1 15.06 84.94 9.80
2014; Oltenacu, 2015; Bezdadea Cătuneanu et
V4.1 14.97 85.04 7.80
al., 2017), with refractive device Kruss DR301-
V5.1 15.60 84.40 7.60
95 (% Brix).
Dry matter and water content of the samples V6.1 17.02 82.98 9.70
were determined by oven drying for 24 hours at V7.1 14.66 85.34 8.10
105oC using a UN110 Memmert oven, method V8.1 15.09 84.91 7.90
used also by Moura (2005), Skupień (2006), Average Vn.1 16.01 83.99 8.54
Delian (2011), Corollaro (2014), Mureşan V1.2 15.51 84.49 8.00
198
Dry matter Water Total Soluble (85.04%). In the wool mulch variants the
Variant content content (U) Solids (TSS)
(D.M. %) (%) (% Brix) minimum was in V8.2 (82.03%) and the
V2.2 17.16 82.84 10.30 maximum in V3.2 (84.84%). In the control
V3.2 15.16 84.84 8.20 mulch variants the minimum was in V7.3
V4.2 17.43 82.57 8.50 variant (81.37%) and the maximum in V4.3
V5.2 16.47 83.53 7.80
variant (86.54%).
V6 variant registered the lowest value of water
V6.2 17.50 82.50 8.70
content in average (82.56%) and V4 variant the
V7.2 15.83 84.17 8.10
maximum value (84.71%).
V8.2 17.98 82.03 9.80
Average Vn.2 16.63 83.37 8.68 Total soluble solids for all the ʻCrown Princess
V1.3 15.75 84.25 8.00 Margaretaʼ variants were in average at 8.64 (%
V2.3 15.24 84.76 7.80 Brix).
V3.3 16.70 83.30 9.60 In the wood chips mulch variants the minimum
V4.3 13.47 86.54 10.20
were in V5.1 (7.60%), V4.1 (7.80%) and V8.1
(7.90%); the maximum values were in V3.1
V5.3 15.98 84.02 9.70
(9.80%) and V6.1 (9.70%).
V6.3 17.82 82.18 7.80
In the wool mulch variants the minimum were
V7.3 18.63 81.37 7.60 in V5.2 variant (7.80%) and the maximum in
V8.3 14.88 85.12 9.00 V2.2 (10.30%) and V8.2 (9.80%).
Average
16.06 83.94 8.71 In the control mulch variants the minimum
Vn.3.
Average were in V7.3 (7.60%), V2.3 (7.80%) and V6.3
16.23 83.77 8.64
Vn.n. (7.80%); maximum were in V4.3 (10.20%),
V5.3 (9.70%) and V3.3 (9.60%).
Water content was similar in all variants, with V7 variant registered the lowest value of total
an average of 83.77%. In the wood chip mulch soluble solids in average (7.93%) and V3
variants the minimum was in V1.1 (80.67%) variant the maximum values of TSS (9.20%).
and the maximum in V6.1 (85.34%) and V4.1
3,50
3,00
2.65
2.47
μg.(ml of plant extract)-1
2.42
2,50
2.22 2.21
2.00 1.95
2,00 1.88
1.78
1.65 1.71
1.63
Wood chips
1.47 1.51
1,50 1.27 1.30
1.37 Wool
1.25
1.25 1.25
1.24
1.13 1.12
0.96 0.93
105
0.96
1.07
0.93
Control
1,00 0.84
0.76
0.71
0.62
0.68
0.62 Average/mulch
0.53
0,50
0,00
Figure 1. Variation of total carotenoids content (μg (ml of plant extract)-1) in fresh petals of ʻCrown Princess Margaretaʼ
variety, according to mulch and ameliorative species variants
199
Total carotenoids content in fresh petals for all Table 3. Influence of variants on evolution of total
ʻCrown Princess Margaretaʼ variants were in soluble solids and dry matter on fresh petals of ʻFalstaffʼ
variety
average at 1.37 (μg.(ml of plant extract)-1).
In the wood chips mulch variants the minimum
Dry matter Water Total Soluble
were in V5.1 (0.62), V7.1 (0.68), V8.1 (0.62); Variant content content Solids (TSS)
the maximum values were in V3.1 (2.65) and (D.M. %) (U) (%) (% Brix)
V2.1 (2.47).
In the wool mulch variants the minimum were V1.1 16.06 83.94 7.20
in V5.2 variant (0.71%) and the maximum in V2.1 13.68 86.32 8.00
V8.2 (2.21%). V3.1 14.63 85.37 8.30
In the control mulch variants the minimum V4.1 19.87 80.13 8.30
were in V4.3 (0.53) and the maximum were in V5.1 15.20 84.80 10.30
V3.3 (2.42%). V6.1 18.59 81.41 9.90
V4 variant registered the lowest value of total
V7.1 23.15 76.85 10.00
carotenoids content in average (0.84) and V3
V8.1 13.31 86.69 8.70
variant the maximum values (2.00) (Figure 1).
Average Vn.1 16.81 83.19 8.84
Table 2. Total soluble solids content, dry matter and total
carotenoids content on dehydrated rose petals and rose V1.2 15.09 84.91 8.40
petal jam from ‘Crown Princess Margareta’ variety V2.2 16.35 83.65 8.00
Total V3.2 15.91 84.09 10.10

Dry
Soluble Total
matter Water V4.2 16.70 83.30 7.40
Solids carotenoids
Variant content content
(TSS) (μg.(ml of plant V5.2 15.25 84.75 8.80
(D.M. (U) (%)
(% extract)-1)
%) V6.2 14.88 85.12 11.10
Brix)
U 91.90 8.10 2.07±0.170 V7.2 13.40 86.60 8.50
D1 20.00 80.00 68.50 0.13±0.082 V8.2 14.33 85.67 7.10
D4 26.00 74.00 61.30 0.05±0.055 Average Vn.2 15.24 84.76 8.68
D7 20.00 80.00 68.10 0.09±0.008 V1.3 14.70 85.31 7.30
Average
22.00 78.00 65.97 0.09±0.063 V2.3 13.19 86.81 6.70
D.n
V3.3 14.34 85.66 7.90
In the dry petals of ʻCrown Princess Margaretaʼ V4.3 14.38 85.62 8.50
variety, total carotenoids were similar to the V5.3 15.78 84.22 7.70
average of fresh petals (2.07±0.170) (Table 2). V6.3 13.96 86.04 7.80
In all rose petal jams from this rose variety, V7.3 13.86 86.14 6.90
total carotenoids were significant lower than in
V8.3 15.09 84.91 7.80
fresh or dry petals.
Average Vn.3 14.41 85.59 7.58
For the ʻFalstaffʼ variety total soluble solids, Average Vn 15.49 84.51 8.36
dry matter and total anthocyanin content in
fresh petals, dry petals and jam were analysed V4 variant registered the lowest value of water
comparing the variants of ameliorative species content in average (83.02%) and V8 variant the
and mulch used (Tables 3, 4). maximum value (85.76%).
Water content was similar in all variants, with Total soluble solids for all the ʻFalstaffʼ
an average of 84.51%. In the wood chip mulch variants were in average at 8.36 (% Brix). In
variants the minimum was in V7.1 (76.85%) the wood chips mulch variants the minimum
and the maximum in V8.1 (86.69%). In the was in V1.1 (7.20%) and the maximum was in
wool mulch variants the minimum was in V4.2 V5.1 (10.30%) and V7.1 (10.00%). In the wool
(83.30%) and the maximum in V7.2 (86.60%). mulch variants the minimum was in V8.2
In the control mulch variants the minimum was variant (7.10%) and the maximum in V6.2
in V5.3 (84.22%) and the maximum in V2.3 (11.10%).
(86.81%).
200
In the control mulch variants the minimum V2 variant registered the lowest value of total
were in V2.3 (6.70%), V7.3 (6.90%) and soluble solids in average (7.57%) and V6
maximum was in V4.3 (8.50%). variant the maximum values of TSS (9.60%).
45,00
40,00 38.74 38.41

37.85
38.49 38.69
36.03 36.35
36.10 36.59 36.15
35.39 35.71
34.26 34.47 34.54
35,00 33.04 33.51 33.16 33.06
32.46
32.42
31.78 31.40 31.83 31.48
30.72 30.31
30.09 30.06
29.6229.08
29.39
30,00 28.56 28.79
26.28
25.26
mg/100 g
25,00
Wood chips
20,00 Wool
Control
15,00
Average/mulch
10,00
5,00
0,00
Figure 3. Variation of total anthocyanin content (mg/100 g) of ʻFalstaffʼ variety,

according to mulch and ameliorative species variants
Total anthocyanin for all the ʻFalstaffʼ variants Table 4. Total soluble solids, dry matter and total
were in average at 33.06 mg/100 g. anthocyanin on dehydrated rose petals and rose petal jam
of ʻFalstaffʼ variety
In the wood chips mulch variants the minimum
was in V3.1 (15.26) and maximum was in V5.1 Total
Total
Dry matter Water soluble
(38.49). Variant content content solids
anthocyanin
content
In the wool mulch variants the minimum was in (D.M.) (%) (U) (%) (TSS) (%
(mg/100 g)
Brix)
V7.2 variant (31.48%) and the maximum in Fd 92.40 7.60 - 39.76±0.272
V3.2 (38.74), V8.2 (38.69) and V4.2 (38.41).
D2 21.00 79.00 68.70 6.67±0.009
In the control mulch variants the minimum was
D5 29.00 71.00 58.60 7.20±0.329
in V3.3 (26.28) and maximum was in V5.3
(36.10). D8 27.0 73.00 61.80 5.96±0.399
Average
V7 and V3 variants registered the lowest values D.n
25.67 74.33 63.03 6.61±0.599
of total anthocyanin in average (30.06
respectively 30.09); V5 and V4 variants
registered the maximum values of total CONCLUSIONS
anthocyanin in average (36.35 respectively
36.03). In the dry petals of Falstaff variety, This study aimed to determine and compare
total anthocyanin content was bigger than the different parameters analyzed in rose petals and
average of fresh petals (33.06±0.626). rose jams, as total carotenoids, total
In all rose petal jams from this rose variety, anthocyanins, total soluble solids and dry
total anthocyanin content was significant lower matter influenced by the three varieties of roses
than in fresh or dry petals. and the applied organic growing technologies.
201
Water content was similar in all variants both Bezdadea Cătuneanu I., Bădulescu L., Dobrin A., Stan
for the two rose variety ʻCrown Princess A., Hoza D., 2017. The influence of storage in
controlled atmosphere on quality indicators of three
Margaretaʼ and ʻFalstaffʼ, with an average of blueberries varieties. Scientific Papers. Series B,
84.14%. Total soluble solid content varied Horticulture. Vol. LXI, 91-100.
with the mulch variants (wood chips and wool) Corollaro M. L., Aprea E., Endrizzi I., Betta E., Demattè
and also with the plant ameliorative species. M.L., Charles M., Bergamaschi M., Costa F., Biasioli
The lower values was in V7 (7.93%) variant F., Grappadelli L.C., Gasperi F., 2014. A combined
sensory-instrumental tool for apple quality
and the biggest in V3 for ʻCrown Princess evaluation. Post-harvest Biology and Technology 96:
Margaretaʼ (9.20%). The maximum value was 135-144.
obtain in V4.3 with 10.20%. For the ʻFalstaffʼ Delian E., Petre V., Burzo I., Bădulescu L., Hoza D.,
the lower value was in V2 variant (7.57%) and 2011. Total phenols and nutrients composition
the biggest in V6 (9.60%). The maximum value aspects of some apple cultivars and new studied
breeding creations lines grown in Voineşti area –
was obtain in V6.2 with 11.10%. Both rose Romania. Romanian Biotechnological Letters, Vol.
varieties have close TSS values on average. 16, No.6: 6722-6729.
Total carotenoids content in fresh petals for all Lambraki M., 2001. Plantes, herbees, Legumes verts,
ʻCrown Princess Margaretaʼ variants was Fruits, 80.
influenced by the mulch and ameliorative Lichtenthaler H.K., Wellburn A.R., 1983.
Determinations of total carotenoids and chlorophylls
variants. V4 variant, Sinapis + Tagetes a and b of leaf extracts in different solvents.
registered the lowest value of total carotenoids Biochemical Society Transactions, 11 (5): 591-592.
content in average (0.84) and V3, Phacelia, Milică C., Bavaru A., Bavaru E., 2010. Trandafirul:
variant the maximum values (2.00). Wood chip gingășie, frumusețe, sănătate. Monografia genului
mulch variants had a positive influence, total Rosa, Editura Pim, Iași, 225-229.
Moura C., Masson M., Yamamoto C., 2005. Effect of
carotenoids content was higher in these osmotic dehydration in the apple (Pyrus malus)
variants. Dried petals preserved the varieties ʻGalaʼ, ʻGoldʼ and ʻFujiʼ. Thermal
carotenoids contents while in the rose petal jam Engineering, vol. 4: 46-49.
was very low. Total anthocyanin content in Mureşan E., Muste S., Borşa A., Vlaic R., Mureşan V.,
fresh petals of ʻFalstaffʼ variants were in 2014. Evalution of physical-chemical indexes, sugars,
pigments and phenolic compounds of fruits from
average at 33.06 mg/100 g, more than in many three apple varieties at the end of storage period.
fruits as blueberry, apple (Bezdadea Cătuneanu Bulletin UASVM Food Science and Technology 71
et al., 2017). V7, Tagetes, and V3, Phacelia, (1).
variants registered the lowest values of total Oltenacu N., Lascăr E., 2015. Capacity of maintaining
anthocyanin in average (30.06, respectively the apples quality, in fresh condition-case study.
Scientific Papers Series Management, Economic
30.09) while the mixed species variants V5 and Engineering in Agriculture and Rural Development
V4 registered the maximum values of total Vol. 15: 331-335.
anthocyanin in average (36.35, respectively Saei A., Tustin D., Zamani Z., Talaie A., Hall A., 2011.
36.03). Wool was beneficial, the maximum Cropping effects on the loss of apple fruit firmness
values on anthocyanin being registered in these during storage: The relationship between texture
retention and fruit dry matter concentration. Scientia
variants. Dried petals preserved the Horticulturae 130: 256-265.
anthocyanins while in the rose petal jam they Skupień K., 2006. Chemical composition of selected
are very low. cultivars of highbush blueberry fruit (Vaccinium
corymbosum L.). Folia Horticulturae 18/2: 47-56.
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F., Zadak Z., 2015. Sugar composition of apple
cultivars and its relationship to sensory evaluation.
Arnon D.I., 1949. Copper enzymes in isolated Nauka. Technologia. Jakość 4 (101): 137-150.
chloroplasts. Polyphenoloxidase in Beta vulgaris. Yoon K. Y., Woodams E.E., Hang Y.D., 2005.
Plant Physiol., 24 (1): 1-15. Relationship of acid phosphatase activity and
Austin D., 2012. Handbook of Roses - European edition, Brix/acid ratio in apples. Lebensm.-Wiss. u.-Technol,
p. 12, 40-41. 38: 181-183.
Barros L., Carvalho A.M., Ferreira I.C.F.R, 2011. Exotic Vasilca Mozăceni A., 2002. Incursiune in Fitoterapie,
fruits as a source of important phytochemicals: Editura Polirom, Iași, 284.
Improving the traditional use of Rosa canina fruits in Wagner Ș., 2010. Trandafiri cu parfum, Editura Napoca
Portugal, Food Research International. Star, Cluj-Napoca, 76-80.
202
CULTIVATION TECHNOLOGY OF ORGANIC ROSES

FOR PETAL PRODUCTION
Ana Cornelia BUTCARU, Florin STĂNICĂ
University of Agronomic Sciences and Veterinary Medicine of Bucharest, Research Centre for
Studies of Food and Agricultural Products Quality, 59 Marasti Blvd, District 1, Bucharest, Romania
Abstract
The aim of this paper is to present a technology for the cultivation of organic edible roses. The basic principle followed
is the creation of an organic production ecosystem considering the natural and anthropogenic factors. To increase the
soil biologic activity and for soil disinfection and disinsection, three ameliorative species: Sinapis alba L., Phacelia
tanacetifolia L. and Tagetes patula L. were used before and after planting. Microbiological and agrochemical analyses
were made to monitor the soil activity and the influence on the rose plants. On the row, the soil was mulched with wood
chips and wool and for irrigation, a drip systems was used. The rose climbing plants were supported by wire trellis. For
plant protection, a strategy to alternate copper and sulfur based products with alternative ones like sodium
bicarbonate, pepper, garlic and propolis tincture and other products based on plant and animal extracts (Mimosa sp.,
chitosan) was followed. Different bio-stimulators and caw milk were used to increase the plant immunity system. The
cultivation technology of organic roses can be implemented on large scale farms with good economical results.
Key words: soil management, plant canopy, fertilization, plant protection, picking, storage.
INTRODUCTION climbing varieties: ʻFalstaffʼ, ʻBrother Cadfaelʼ

and ʻCrown Princess Margaretaʼ, was
Rose is one of the most mankind beloved plant established (Austin, 2012).
specie since the ancient times. It is a symbol of The rose crop was protected by planting a
love and victory, sign of nobility and poplar (Populus balsamifera) wind break.
refinement. Its therapeutic properties are At the beginning, to increase the soil biologic
usually forgotten in favor of the ornamental activity and for soil disinfection and
role (Lambraki, 2001; Milică et al., 2010; disinsection, three ameliorative species: Sinapis
Vasilva Mozăceni, 2002). It is important to alba L., Phacelia tanacetifolia L. and Tagetes
rediscover them and for an appropriate analyze patula L. were used before and after planting.
an organic culture is needed. Microbiological and agrochemical analyses
The aim of this paper is to present a technology were made to monitor the soil activity and the
for the cultivation of organic roses for petal influence on the rose plants (Butcaru et al.,
production. 2016; Butcaru et al., 2017)
The basic principle followed is the creation of On the row, the soil was mulched with saw dust
an organic production ecosystem considering and wool and for irrigation, a drip systems was
the natural and anthropogenic factors (Milică et used (Matei et al., 2017; Butcaru et al., 2017).
al., 2010; Wagner, 2010). The inter-row was kept grassy through repeated
The cultivation technology of organic roses for mowing.
petal production detailed in this paper, can be The rose climbing plants were supported by
implemented on large scale farms with good wire trellis and starting with the second
economical results. growing year, a specific pruning scheme was
applied. Each plant was formed with three
MATERIALS AND METHODS parallel, vertical shoots (Trident). In the first
year, from the beginning of flowering, the
At the experimental field of the University of flower buds were eliminated until the principal
Agronomic Sciences and Veterinary Medicine shoots reached more than 1 m height. After
of Bucharest, an organic rose culture with three that, the roses were left to flower.
203
The same organic crop management regarding 1.2. Plotting the land
fertilization and plant protection was applied Depending on the size of the land taken in the
for all experimental variants. Manure was crop, it can be divided into several plots for
applied at planting, while other organic ease of maintenance. Usually the parcels are
fertilizers on different phenological stages. rectangular, the rows of roses being parallel to
Wool mulch applied on plant rows proved to the small side of the plot. At the same time,
have high qualities as organic fertilizer too. roads and return areas are usually arranged,
(Butcaru et al., 2017) which are usually ground or sown.
For plant protection, a strategy to alternate
copper and sulfur based products with 1.3. Fencing the culture
alternative ones like sodium bicarbonate, Fencing the rose culture is intended to protect
pepper, garlic and propolis tincture and other against wild animals. It is possible to use a
products based on plant and animal extracts galvanized net, stretched on concrete pillars,
(Mimosa sp., chitosan) was followed. Different 2.5 m high, fixed at 2.0 m between them. From
bio-stimulators and caw milk were used to 50 to 50 m the pillars will be provided with
increase the plant immunity system. oblique counter blades in order to stretch the
The petals were collected in plastic bags during corresponding net.
the full flowering period (from May to The net will have a height of 2.0 m. Its base
October), in the morning (7.00 - 10.00 a.m.). has to be buried 10 to 15 cm in the ground to
After sorting and cleaning, petals were used for prevent the rabbits or other animals from
drying or processing as jam. The unprocessed entering under it.
petals were kept in cold storage at 1-20C and
85-95% humidity for 7-10 days. Diferent rose 1.4. Creating the windbreak
petal jam variants were tested for production The organic edible rose culture is well suited to
(Butcaru et al., 2017). having a windbreak both in creating a
microclimate where external influences are
RESULTS AND DISCUSSIONS diminished (possibly polluting substances from
neighboring crops) and to create a shelter for
The technology for the cultivation of organic useful avifauna.
roses for petal production consists in several In the established culture within the
stages, presented below. Experimental Field of UASMV Bucharest, a
poplar windbreak (Populus balsamifera) was
Stage 1. Preparing the land for planting planted (Figure 2). In March 2015, a grass band
1.1. Land clearing of existing vegetation and 2.0-2.5 m wide was sown on all four sides of
clean the other existing rubbish the cultivated land (1,350 m2). In December
This operation consists in cleaning the land 2015, the boreholes were dug and planted
from the older vegetable debris of the last poplar (Populus balsamifera) on each side, 1 m
culture and in the same time eliminates stones, apart, respectively, in the center of the strip.
boulders, glass etc. (Figure 1).
Figure 2. Leveling band with drainage slope; Poplar

windbreak (Populus balsamifera)
1.5. The proper preparation of the land

1.5.1. Scarification of the land
Scarification of the land is done in two
directions, with a scarifier with active organs at
Figure 1. Collecting debris (stones, boulders, glass)
204
a depth of 60-80 cm. It has the role of
deflecting the land in depth and removing any
roots from the ground. It is good to be done a
few months ahead.
1.5.2. Ground leveling

It can be done either by plant soil intake or by
leveling the existing one. In the land taken in
culture, in March 2015, the small oscillation of
level were leveled by moving a small amount
of surface soil vegetation between different
points.
1.5.3. Depth plowing 25-30 cm Figure 3. Ameliorative plants details with soil
It is usually done in autumn with specific incorporation
machines.
Stage 2. Establishing the organic edible rose
1.5.4. Soil preparation with a rotary cutter culture
It has the role of creating a perfectly flat 2.1. Establishing the culture
surface with well-grounded soil. 2.1.1. Supply of seedlings
In order to establish an organic rose culture,
1.5.5. Natural soil disinfection and seedlings must be procured in advance. In
organic farming, these must be certified. If
disinsection
organic suppliers of organic planting material
An important stage in the soil preparation in an are not available, conventional seedlings can be
organic system consists in the actions carried used with the proving the absence of certified
out in order to increase the biological activity seedlings. If the plants arrive long before
in the soil. One step in this direction is the planting, they are stratified into ditch or
sowing of ameliorative plants with a role in soil containers.
restructuring, the elimination of dangerous soil
diseases and pests, the potentiation of fungi and 2.1.2. Establishment of planting distances and
favorable soil bacteria (Figure 3). It is land plotting
recommended to use the herbaceous plant Depending on the varieties of roses to be
species: mustard (Sinapis alba) at 12 kg/ha, planted, the distance between the rows and the
Tagetes sp., 6-8 kg/ha, Phacelia tanacetifolia in row is then determined, followed by picketing
the amount of 8-10 kg/ha. They are sown early on the field.
spring (March - early April). After blossoming For the used varieties of roses, planting
(50 to 55 days for mustard and Phacelia; 70 distances were 2 m between rows and 1 m per
days for Tagetes sp.) the plants are row. To make the plotting, pickets of 0.5 m
incorporated in the soil. length, 50 m long roulette and 0.5 m long
The best results on the potentiation of knotted wire of about 50 m long are used.
microbiological activity in soil were observed When land is picked up, it is meant to mark the
when plants were used in combinations of 2 or place where each rose will be grown. Direction
3 species. Since they have different flowering of rows is recommended to be north - south.
periods, sowing is recommended in strips 1-3 Considering the 1 m distance between plants, it
meters wide. is recommended to open planting ditches.
After incorporation of the plants into the soil,
the soil is kept free of weeds until the 2.1.3. Basic fertilization at planting
establishment of the crop, by mowing them and After opening the planting ditch, well
leaving them on the ground. fermented manure (10-15 t/ha) is applied. Other
fertilizers in organic farming can also be
applied.
205
In areas where it is difficult to purchase
manure, it can be replaced by other types of
fertilizer such as Organofert (based on poultry
manure) (2 t/ha).
2.1.4. Installing trellis system

The trellis system is usually installed prior to
planting. The variant chosen for the crop was
made up of: acacia pillars and counter pillars of
diameters 80-100 cm, height 300 cm and pillars
for interior of diameter 60-80 cm, height 300
cm. The pits can be done with the help of the Figure 5. Fertigation system details
motor or manual drill 100 cm (Figure 4).
The pillars are fixed to the rows of roses at a 2.1.6. Rose planting
distance of about 7 m between them and at a Planting period. The most suitable period for
depth of 60 cm. It is first burned at the base planting is autumn after the fall of the leaves.
with the burner at a length of 1 m. Preparing plant material. The plants have to be
Three galvanized wires of 2.5 mm are fastened hydrate 24 hours before the planting time.
with bows, the first one at 70 cm of soil, the Check the condition of the roots, where dry or
second one at 130 cm and the third one at 200 injured portions are eliminated until to the
cm of soil. The second wire is recommended to healthy portion. Then the roots are dipped in a
be doubled to direct the branches of roses mud made from fresh dung, to which the
inside the created space. addition of bio stimulators for root formation
(eg Rootip mix 2-3 l/ha) is recommended.
Planting. The plants are planted with the
grafting point located 10-15 cm above ground
level. Place it with its roots in the planting ditch
on a loose soil mound. Draw soil from the soil
fertile ground; rotate evenly around the plant
from the outside to the inside.
An organic-mineral fertilizer can be applied at
this time to stimulate the rapid growth of the
roots or later to a few days application of
Rootip mix through the fertilization system.
Then the rest amount of soil is added and wet
with at least 15-20 liters of water/plant.
In the case of autumn plantings, the roses are
then mussed by pulling the ground at the base
of the roots, forming a 20-30 cm high hill.
Early spring (end of February - beginning of
March) breaks down to favor growing buds.
Figure 4. Trellis system details
2.1.5. Install fertigation system

The fertigation system consists of a main
pipeline and several secondary pipes for each
row of roses. The drip tubes were lifted on the
first wire of the support system (Figure 5).
Figure 6. Planting details
206
2.1.7. Soil maintenance Remove the blooms until the bush reaches an
It is recommended to mulch the roses. Wool, average height of 1m, and then let it bloom.
wood chips or different wraps can be used. As a form of branch leadership, it can be
Wool has the advantage of being a very good chosen three vigorous branches that make up
fertilizer. It is recommended to use it in a the skeleton elements, the rest shortening.
slightly processed form to avoid the holes that Year 2. In the second year, the three branches
leave the weeds to grow (Figure 7). of the previous year remain, springing down
the anticipated branches. Shorten to the ring or
very short the other branches of the bush
(Figure 8).
Figure 7. Mulch details (wool and wood chips)
Between rows, the vegetation is regularly

mowed, keeping it at low heights.
2.2. Training pruning

Year 1. Early spring pruning works are carried Figure 8. Pruning details (first and second year)
out, following the development of the bushes
and the distribution of uniform branches.
2.3. Organic treatment

Table 1. Scheme of organic treatments for edible roses
No. Phenophase Pathogen/pest Recommended plant protection products
1 The beginning of vegetation Aphids, mites (hibernate forms) - vegetable oil (Ovipron 2000 - 0,5%)
- Oleorgan – 0,3%
- Laser 240 – 0,25%
- Canelys – 0,3%
Mildew, Black spot, Rust - Bouille bordelaise – 0,5-1,0%
- Microthiol – 0,3%
- sodium bicarbonate – 0,5%
- Mimox – 0,3%
- Altosan – 0,3-0,8%
2 Growing shoots (April - May) Diseases produced by fungi and - Bouille bordelaise – 0,5%
bacteria (Black spot, mildew, - Microthiol – 0,3%
rust) - sodium bicarbonate – 0,5%
- Mimox – 0,3%
- Altosan – 0,3-0,8%
Aphids, other pests - Oleorgan – 0,3%
- Laser 240 – 0,25%
- Canelys – 0,3%
3 June - September Diseases produced by fungi and - Bouille bordelaise – 0,5%
bacteria (Black spot, mildew, - Microthiol – 0,3%
rust) - sodium bicarbonate – 0,5%
- Mimox – 0,3%
- Altosan – 0,3-0,8%
Aphids - Oleorgan – 0,3%
- Laser 240 – 0,25%
- Canelys – 0,3%
- propolis tincture – 0,15%
- hot pepper tincture – 0,3%
-garlic tincture – 0,3%
4 Fall of the leaves (November - Mycotic pathogens Bouille bordelaise – 0,5% - 1,0%
December) and bacteria
5 Winter period Mycotic pathogens - vegetable oil (Ovipron 2000 - 0,5%)
and bacteria, pest (hibernating - Bouille bordelaise – 1,0%
forms)
207
2.4. Fertilization
Table 2. Scheme of fertilization for the organic edible rose culture
No Phenophase Recommended plant protection products
1 Înfrunzit, creşterea lăstarilor (aprilie - mai) Alga 0,3% (foliar or radicular)
Magnetic fertilizer
Cropmax 0,2%
Caw milk 1,0%
2 June - September Alga 0,3% (foliar or radicular)
Cropmax 0,2%
Caw milk 1,0%
3 At the fall of the leaves, during the winter period Bentonite 1,5% (frost protection)
2.5. Rose petal harvest

In the first year, the harvesting of flowers
begins in July, after the plants have reached an
average height of 1 m. Since the second year,
the flowers can be harvested since May (Figure
9).
Petals are harvested in the morning, after dew,
in pots or baskets. It is sorted immediately,
eliminating the various impurities.
They are packaged in vacuum bags of
convenient size for the assigned destination
(according to the specific quantities).
The vacuum bags are kept in a cold store at
Figure 9. Flowering details (second year)
temperatures of 1-2°C when the petals are
processed for the next 5-7 days or in the freezer
at temperatures of -18 to -20°C for extended
periods.
Table 3. Technology to establish an organic edible rose culture
No. Operațion Period Equipment / Supplies
1 Land clearing of existing vegetation and March- September, year I Backhoe/bags /
clean the other existing rubbish. Containers/Tractor with trailer
2 Plotting March- September, year I Pickets, roulette
3 Fencing March- September, year I Drilling tractor/Concrete pillars (h = 2.5 m),
galvanized mesh (h = 2 m)
4 Land scarification March- September, year I Scarifier
5 Ground leveling March- September, year I Backhoe
6 Depth plowing 25-30 cm September – October, year I Tractor
7 Soil preparation with a rotary cutter February – March, year II Tractor, rotary cutter
8 Creating the windbreak March- September, year I Drilling tractor/Poplars (Populus balsamifera)
manure
9 Natural soil disinfection and March– April, year II Sowing equipment/ seeds: mustard (Sinapis alba)
disinsection - 12 kg/ha, Tagetes sp. - 6-8 kg/ha, phacelia
(Phacelia tanacetifolia) - 8-10 kg/ha.
Tractor
10 Supply of seedlings September, year II Seedlings
11 Establishment of planting distances and October – November, Pickets, roulette, string
land plotting year II
12 Installing trellis system October – November, Drill 100 mm/ acacia pillars, burner, wire 2,5 mm,
year II staples, wire tensioners
13 Install fertigation system October – November, Principal pipelines, drip, elements
year II
14 Opening the planting ditch October – November, Tractor
year II
15 Basic fertilization at planting October – November, Fertilization distribution equipment /
year II manure (10-15 t/ha) or other (ex. Orgafert 2 t/ha)
16 Rose planting October – November, scissors/fresh dung mud/Rootip mix (2-3 l/ha)
year II
208
Table 4. Maintenance of an organic rooting plant
No. Operation Period Equipment/Supplies
1 Winter treatment January- February - vegetable oil (Ovipron 2000 - 0,5%)
- Bouille bordelaise – 1,0%
- bentonite 1,5% (frost protection)
2 Eliminate the winter soil hills February - March - dig/ dibble
3 Pruning and trellising branches February - March - Scissors/trellis elements
4 Eliminate dried leaves, branches (cultural March - Scissors
hygiene)
5 Spring treatment March Insecticides:
- Oleorgan – 0,3%
- Laser 240 – 0,25%
- Canelys – 0,3%
Fungicide/Bio-stimulators:
-Bouille bordelaise – 0,5-1,0%
- Sodium Bicarbonate – 0,5%
- Mimox – 0,3%
- Altosan – 0,3-0,8%
6 Soil maintenance March - Wool/twood chips/ processed wool
- trimer/ mowing equipment
7 Ameliorative plants sowing March – April -Sowing equipment/ ameliorative plants seeds
8 Treatment sprout growing and fertilization April – May Fungicide/Bio-stimulators
- Mimox – 0,3%
- Altosan – 0,3-0,8%
Insecticides
- Oleorgan – 0,3%
- Laser 240 – 0,25%
- Canelys – 0,3%
Fertilizers
- Alga 0,3% (foliar or radicular)
- Magnetic fertilizer
- Cropmax 0,2%
- Cow milk 1,0%
9 Remove the blooms (year I) May – June
10 Soil maintenance May – October - trimer/ mowing equipment
11 Irrigation May – September Irrigation system
12 Trellising new sprouts June – September - trellising elements
13 Protection and fertilization treatments June – September Fungicide/Bio-stimulators
- Mimox – 0,3%
- Altosan – 0,3-0,8%
Insecticides
- Oleorgan – 0,3%
- Laser 240 – 0,25%
- Canelys – 0,3%
- propolis tincture – 0,15%
- hot pepper tincture – 0,3%
-garlic tincture – 0,3%
Fertilizers
- Alga 0,3% (foliar or radicular)
- Cropmax 0,2%
- Caw milk 1,0%
- Bombardier 0,25%
14 Rose petals harvesting July - October - Boxes/baskets/scissors
15 Complete plants October - December - drill, scissors/manure/fresh dung/
seedlings/Rootip mix
16 Falling leaves treatments November - - Bouille bordelaise – 0,5% - 1,0%
December
209
The study aimed to present the technology for Austin D., 2012. Handbook of Roses - European edition,
an organic edible rose crop. It had, as basic 12, 40-41.
Butcaru A.C., Stănică F., Madjar R., 2017. Influence of
principle, the creation and maintenence of an ameliorative plants and mulch on some soil
organic production ecosystem considering the agrochemical characteristics in an organic edible rose
natural and anthropogenic factors. crop, USAMVB Scientific Papers, Series
The technology focus in the first stages of Horticulture, Vol. LXI, 101-106.
establishment of the organic edible rose crop on Butcaru A.C., Stănică F., Matei G.M., Matei S., 2017.
Soil microbial activity in an organic edible rose crop,
the increase of soil biological activity. USAMVB Scientific Papers, Series Horticulture, Vol.
Using three ameliorative species (Sinapis alba, LXI, 107-112.
Phacelia tanacetifolia and Tagetes patula) the Butcaru A.C., Stănică F., Velcea M.N., 2017.
soil bacteria and fungi increased at important Preliminary studies regarding the production of jam
levels. from organic rose petal, Bulletin UASVM Food Science
and Technology 74 (2)/2017 ISSN-L 2344-2344;
Climbing rose varieties used proved several Print ISSN 2344-2344; Electronic ISSN 2344-5300
advantages towards the classical edible rose: DOI: 10.15835/buasvmcn-fst: 0022, 50-57.
trellising on the vertical position, repeated Butcaru A.C., Stănică F., Matei G.M., Matei S., 2016.
flowering since May to October and fragrance Alternative methods to improve soil activity before
varieties with multiple way of valorisation. planting an organic edible rose crop, Journal of
Horticulture, Forestry and Biotechnology, Volume 20
An innovative pruning system refer to a 2 year (4): 12-17, ISSN: 2066-1797.
rotation scheme of branches, giving the Lambraki M., 2001. Plantes, herbees, Legumes verts,
possibility to have a constant production every Fruits, 80.
year. Matei G.M., Dumitru S., Matei S., Butcaru A.C., Stănică
The average quantity of petals per hectare was F., 2017. Microbiological aspects in soil microbiome
composition and activity under the influence of
9.44 t/ha at ʻCrown Princess Margaretaʼ organic mulches, Conference Proceedings Vol. 17,
variety, 7.54 t/ha at ʻFalstaffʼ variety and 5.62 Nano, Bio, Green and Space-Technologies for a
t/ha at ʻBrother Cadfaelʼ variety. Sustainable Future Issue 61, Micro and Nano
The best variant of mulch used was wool, Technologies, Advances in Biotechnology, 813-820.
which proved to have special qualities as a Milică C., Bavaru A., Bavaru E., 2010. Trandafirul:
gingășie, frumusețe, sănătate. Monografia genului
fertiliser too. Rosa, Editura Pim, Iași, 225-229.
The best temperature for maintain the harvested Vasilca Mozăceni A., 2002. Incursiune in Fitoterapie,
petals were 1-2°C when the petals are Editura Polirom, Iași, 284.
processed in the next 7-10 days. Wagner Ș., 2010. Trandafiri cu parfum, Editura Napoca
Star, Cluj-Napoca, 76-80.
210
MORUS SP. FOR REVIGORATING SILKWORM BREEDING

IN ROMANIA AND PROMOTING HEALTH BENEFITS
OF LEAVES AND FRUITS
Otilia BOBIS1, Daniel Severus DEZMIREAN2, Liviu Alexandru MĂRGHITAŞ2,

Victoriţa BONTA1, Adriana URCAN2, Claudia PAŞCA2, Adela Ramona MOISE2
1
Life Science Institute “King Michael I of Romania”, University of Agricultural Sciences
and Veterinary Medicine Cluj-Napoca, 3-5 Mănăştur st. Cluj-Napoca, Romania
2
Faculty of Animal Breeding and Biotechnology, University of Agricultural Sciences
and Veterinary Medicine Cluj-Napoca, 3-5 Mănăştur st., Cluj-Napoca, Romania
Abstract
Besides supplying food for silkworm rearing, initiation of a mulberry plantation is an important business, which may
bring important income in rural areas. Plants of the genus Morus are known to be a rich source of bioactive
compounds from flavonoids and phenolic acids, compounds with known antioxidant and other benefic properties. The
aim of the present study was the characterization of mulberry extracts (from leaves and fruits) regarding their chemical
composition and determination of some bioactive properties of the mentioned matrices. Sugar profile, total protein and
lipid content, total phenolics and flavonoids, mineral composition and aminoacid profile were determined. Mulberry
leaves are important protein sources (being the principal feed for silkworm) and also important source of free
aminoacids, including the essential ones. 25 free aminoacids (aa), were identified and quantified by internal standard
method using LC-MS and EZ: faast Phenomenex kit. The main aa were alanine (144 mg/100g) and Gama-aminobutiric
acid (153 mg/100g). The main minerals from mulberry extracts were: iron, zinc and calcium. Total identified simple
sugars ranged between 8 and 13% and lipid content between 0.5 to 1.5%.
Key words: Morus sp., agriculture, mulberry leaves; bioactive properties, silkworm
INTRODUCTION properties (Imran et al., 2010; Okatan et al.,

2016), blood glucose reductor (Shin et al.,
Mulberry (Morus sp.) are grown worldwide for 2016), antibacterial, anti-inflammatory and
sericulture. Their leafs being the only natural anticancer activity, as well as cardiovascular
food for silkworms, but also for other purposes, and cardioprotective activity (Gryn-Rynko et
among them being fruit production. The most al., 2016). Even the phytochemical composition
popular species are Morus alba and Morus of leaves and fruits of mulberry are studied
nigra, and the intensive selection and mutation worldwide, new promising sources of natural
breeding have resulted in thousands of culti- antioxidants or other phytotherapeuticals can be
vars, hybrids and polyploids (Yongkang, 2000). found in this valuable matrix.
Mulberry trees are valuable species not only for The aim of our study was to evaluate the
silkworm rearing, but also for gardening and chemical composition of extracts from fresh
landscaping, street shade and reduction of and dry leaves and fruits of Morus nigra,
pollution level in the environment and soil Ukraina variety.
(Rafati et al., 2011).
The fruits are important sources of bioactive MATERIALS AND METHODS
compounds such as simple sugars (glucose and
fructose) (Imran et al., 2010), alkaloids (Kim et Chemicals and materials. All chemicals and
al., 2014), carotenoids, polyphenols, antho- reagents were analytical grade or chromate-
cianins and minerals (Dimitrijevic et al., 2014; graphic grade and ultrapure water, purchased
Okatan et al., 2016). from Sigma, Merck and Fluka.
The bioactivity of mulberry fruits was deter- Mulberry leafs, belonging to Morus nigra
mined in many studies, having antioxidant specie, ʻUkrainaʼ vartiety, were harvested in
211
2017, placed at -18oC or air dried in the dark made with Büchi, KjelFlex K-360 unit, every
and grounded in fine powder and kept at 4oC determination is made with a mixture of
until analysis. Mulberry fruits were harvested reagents (50 ml H2O: 90 ml NaOH: 60 ml
in 2017, and kept at -18oC in plastic bags or H3BO3). Titration is made with automatic
dried in the oven at 60oC until constant weight. titrator TitroLine Eeasy (Schott), using 0.05M
After drying, the fruits were grounded in fine sulphuric acid for low protein matrices, until
powder and refrigerated in plastic bags until pH of 4.65.
determinations. Determination of mineral content. To deter-
Sample preparation. Mulberry fruits and leafs mine the levels of micro and macroelements:
were prepared according to the parameter to be Na, Mg, K, Ca, Fe, Pb from studied plant
determined: water extracts, ethanol extracts or matrices, the atomic absorption spectrometry
calcinations for mineral determination. method was used. The mineralization of the
Sugar spectrum. Method used for determi- samples was performed in a microwave
nation of sugar spectrum is high performance furnace, Berghof digestion system MWS-2.
liquid chromatography with refractive index Approximately 0.3 grams of the homogenized
detection (Bonta et al., 2008). samples were placed in special Teflon tubes, 2
Operational parameters of chromatographic ml of 65% HNO3 was added and let to react for
system were: column: modified Alltima Amino 15 minutes, after which 3 ml of H2O2 was
100Ẳ, 5 μm, 250 x 4.6 mm; mobile phase flow: added before the container was sealed (Quinn
1.3 ml/min; mobile phase: acetonitrile/water et al., 1994; Finger et al., 2014). At the end of
(75/25; v/v); column temperature: 30oC; injec- the initiated program, the solution is transferred
tion volume : 20 μl; separation time: 60 min. into plastic containers and the sample is diluted
Sugar standard solutions are prepared like the with ultrapure water to a volume of 125 ml.
analyzed sample. Standards are injected and An Aanalyst 800 Atomic Absorption Spectro-
analysed separately to determine the retention meter from Perkin-Elmer was used, equipped
time of each sugar and in mixture for the with a cross-linked graphite furnace. It is
calibration curve construction. Results are electrically heated and the voltage will be
expressed in g/100 g sample. applied transversely to the tube, perpendicular
Lipid content. Total lipids were determined by to the light beam, and finally the electromagnet
extraction with organic solvents using Soxhlet will generate a magnetic field parallel to the
method. Two grams of sample were weighted radiation beam emitted by the lamp (Farrukh,
on filter paper, which will be packed and 2012).
placed in the paper cartridge. Dry and clean Free aminoacid profile. Aminoacid profile was
extraction glasses containing 2 boiling stones, determined by liquid chromatography – mass
will be weighted and together with the cartridge detection (LC-MS), as a method with high
and the solvent (70 ml n-hexane) will be fixed selectivity and sensitivity. Determinations were
in PTFE cylinders. The method is set from the made on a Shimadzu LC-MS (Japan), with
multistat: extraction temperature 140°C, electrospray ionization, opedrating in positive
extraction time 3 h, 25 min, washing 30 min, mode. Operational parameters of the method
solvent evaporation in hot air flow 10 min. were: EZ: faast AAA-MS, 250 x 3.0 mm
Extraction glasses, dried in the oven at 60oC chromatographic column, mobile phases:
and cooling, are weighted and the result were ammonium formate 10 mM in water (A) and
expressed as percent. ammonium formate 10 mM in methanol (B),
Protein content. From the homogenized 0.3 ml/min flow, column temperature 35ºC,
sample, 1g is weighted in paper bags handled injection volume 1 µL, detector tension 1.7
with a tweezer so that they are not contami- KV, acquisition time 33 min.
nated with different substances that may con- Polyphenolic and Flavones Contents. For the
tain nitrogen. Paper bags are placed in diges- total polyphenol content determination, the
tion unit vialse (Buchi Digesion Unit K-424), Folin-Ciocâlteu method was used (Folin and
with 2 Kjeldahl tablets and 20 ml concentrated Ciocâlteu, 1927), modified by various authors
sulphuric acid (95-98%). Digestion lasts 2 h, and adapted to all types of matrices (Duda et
until the solution turns to green. Distilation is al., 2015; Mihaylova et al., 2013; Singleton and
212
Rossi, 1965; Kim et al., 2003). A volume of 25 Table 1. Sugar spectrum, total lipids and total
μl of each ethanolic extract was mixed for 5 nitrogen (%) of fruits and leaves of Morus nigra
minutes with 125 μl of 0.2 N Folin-Ciocâlteu. Specification
Fresh
Dry leaves Fresh fruits Dry fruits
leaves
The samples were incubated in the dark for 120 Glucose 0.56 1.48 5.96 27.09
min. The absorbance was measured at 760 nm, Fructose 0.89 1.77 6.16 29.65
using a Sinergy 2 Biotek Multichannel Zaharoza 1.43 3.06 - -
Galactoza 1.02 2.03 - -
spectrophotometer. The standard curve was
Total lipids 2.41 3.40 1.51 3.65
prepared by using different concentrations of Total nitrogen 11.56 19.70 0.96 1.57
gallic acid and the results were expressed as
gallic acid equivalents/100 g). Leaves of mulberry contain fructose and glucose in
The quantification of flavones in the samples small amounts, but a higher content of sucrose.
was made using Dowd method (1959) based on Also galactose is present in this matrix. Instead, the
the reaction of aluminium chloride, as specific fruits contain only fructose (in higher amounts) and
reagent, with the flavonoids present in the glucose. The amounts determined in our study were
in accordance with other studies of Morus nigra
sample giving a yellow color, proportionally
fruits (Okatan et al., 2016). Dry fruits are important
with the concentration of the compounds, simple sugar sources, as seen in Table 1.
determined spectrophotometrically at 415 nm. Leaves of mulberry contain high amounts of lipids,
Radical Scavenging Activity Assay (DPPH). several times higher than determined by different
The antioxidant activity is the primary step in authors (Gryn-Rynko et al., 2016; Imran et al.
determining the biological activity of any 2010), but smaller than those of Iqubal et al. (2012).
natural matrix. DPPH free radical method is an However, the presence of an appreciable content of
antioxidant assay based on electron-transfer lipids demonstrates the potential of these leaves to
that produces a violet solution in ethanol. This have dietary purposes with promising nutritional
free radical, stable at room temperature, is attributes.
reduced in the presence of an antioxidant Higher amounts of lipids were obtained in the fruits
of Morus nigra, compared with literature studies
molecule, giving a discoloration of solution,
(Imran et al., 2010).
proportionally with the amount of antioxidant The contents of some minerals and lead from
present in the sample (Paşca et al., 2016; Duda mulberry leaves and fruits are given in Table 2.
et al., 2015). The free radical scavenging ability
of the ethanolic extracts was measured in terms Table 2. Mineral content of leaves
of hydrogen donation or radical scavenging and fruits of Morus nigra
ability using this method. Thus, 5 μl of 1% Specification
Fresh
Dry leaves Fresh fruits Dry fruits
plant alcoholic extract was mixed with 295 μl leaves
Na (mg/kg) 23.41 50.51 71.65 81.51
of 0.02 mg/ml DPPH solution in methanol, Mg (mg/kg) 2.54 4.03 41.80 56.76
stirred and incubated in the dark for 20 Ca (mg/kg) 542.25 908.79 2693.82 7102.44
minutes. The absorbance changes were Fe (mg/kg) 50.41 76.21 67.52 86.86
K (mg/kg) 1756.22 3308.46 3333.33 7009.18
monitored at 517 nm using a Sinergy 2 Biotek Pb (mg/kg) 0.00 0.00 0.00 0.00
multichanel spectrophotometer. The percent
inhibition of DPPH free radical was calculated The distribution of the elements among the
by the formula: Percentage inhibition (% I) = leaves and fruits of mulberry were in favour of
[(Ablank - Asample)/Ablank] × 100, where, Ablank is fruits, much higher amounts being determined
the absorbance of the control reaction (DPPH in fruits compared to leaves.
alone) and Asample is the absorbance of DPPH No lead was determined in leaves and fruits of
solution in the presence of the test compound. mulberry, indicating no pollution in the area of
mulberry plantation.
RESULTS AND DISCUSSIONS Dimitrijevic et al. (2014) found in mulberry
fruits different heavy metals in fruits of
Mulberry fresh leaves and fresh fruits have a mulberry grown in Serbia (9 mg/kg lead and 36
high content of water (62.07% in leaves and mg/kg nichel).
40.89% in fruits). Very high amounts of potassium were
Chemical composition of leaves and fruits of determined both in leaves and in fruits of
Morus sp. are presented in Table 1. mulberry. If the leaves are good source of pota-
213
ssium for silkworm, the fruits are very good (903.1 mg/100 g), followed by the leaves from
candidates for potassium supplementation in middle (643.2 mg/100 g) and the leaves from
human diet. High amounts of calcium (7102.44 the top of the tree (541.6 mg/100 g).
mg/kg) were determined in dry fruits. This High amount of alanine and Gama-
mineral is very important in diet, and having aminobutiric acid were determined (Table 3),
such a high content in the fruits of Morus followed by asparagine and proline.
nigra, makes this fruit even more valuable. Phenolic compounds are secondary metabolites
Iron and magnesium determined in our study of plants and contribute to different flavours
was similar with other studies (Imran et al., (sweet, bitter, astringent), and determine the
2010). As can be seen in Table 2, mulberry antioxidant activity of the matrix (Thomas-
fruits may be considered as good sources of Barberan and Espin, 2001).
magnesium. The amount of total polyphenols and total
Very different amounts were determined by flavonoids are presented in Figure 1.
Yigit et al. (2010) in some varieties of mulberry
(leaves and fruits) drom Turkey. Similar results
were obtained only for calcium and sodium.
Taking into consideration the high amount of
protein from mulberry leaves, we conducted a
study of determining the free aminoacid profile
from this matrix (Table 3).
Table 3. Aminoacid profile of mulberry leaves
Aminoacid Leaf Leaf Leaf

(base) (middle) (top)
Arginine 33.46 20.48 45.54 Figure 1. Polyphenolic and flavonoid content
Serine 164.43 16.25 17.05 in mulberry leaves and fruits
Asparagine 75.67 43.33 46.63
1-metil-histidine 0.18 0.14 0.21
4-hidroxiproline 46.29 27.25 28.82
Higher amounts of phenolics (total polyphenols
Glicine 13.21 15.07 8.85 and flavonoids) were determined in fruits,
Glicina-proline (dipeptide) 0.28 0.23 0.27 compared to leaves.
Treonine 29.80 31.10 23.61
The obtained results were in accordance with
Alanine 144.01 128.96 66.54
Gama-aminobutiric acid 153.30 129.07 105.78 literature studies (Dimitrijevic et al., 2014;
Alfa-aminobutiric acid 0.32 0.51 0.00 Popescu et al., 2014; Okatan et al., 2016)
Ornitine 0.55 0.38 0.48 Radical scavenging activity of the ethanolic
Metionine 0.21 0.13 0.17
Proline 60.66 47.63 42.90
extracts from leaves and fruits of mulberry
Lizine 23.20 21.49 23.46 ranged between 45 - 67% inhibition percent,
Aspartic acid 11.39 15.36 12.52 similar to literature studies (Iqbal et al., 2012;
Histidine 6.72 6.14 5.64 Dimitrijevic et al., 2014; Okatan et al., 2016).
Thiaproline 0.06 0.08 0.02
Valine 25.69 26.93 12.80
Glutamic acid 27.29 33.07 20.18 CONCLUSIONS
Triptophan 13.58 12.57 12.60
Leucine 24.94 22.75 21.07
Phenilalanine 18.71 16.00 20.12
The high polyphenolic content, together with
Izoleucine 11.45 12.29 8.71 the high amount of simple sugars, total lipids
Tyrozine 17.71 16.07 17.64 and total proteins, underline the nutritive and
phytopharmaceutical potential of mulberry.
Twenty five free aminoacids were detedrmined This study suggested that mulberry fruits, but
in the leaves of mulberry, harvested from three not only, may be used as a potential healthy
different parts of the tree. No significant food, or an important antioxidant carrier for
differences were observed in the three locations different pharmaceutical applications and food
of the tree in respect of free aminoacid profile, supplements.
but the total amount of free aminoacids were
determined in leaves from the base of the tree
214
ACKNOWLEDGEMENTS Mol. Sci., 13, 6651-6664, doi:
10.3390/ijms13066651.
Kim S.B., Chang B.Y., Hwang B.Y., Kim S.Y., Lee
Financial support was granted by the project ADER
M.K., 2014. Pyrrole alkaloids from the fruits of
413/2015, developed through the Ministry of Morus alba. Bioorganic & Medical Chemistry
Agriculture and Rural Development, Romania. The Letters, 24 (24), 5656-5659
authors declare no conflict of interests. Kim D.O., Chun O.K., Kim Y.J., Moon H.Y., Lee C.Y.,
2003. Quantification of polyphenolics and their
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215
216
IN VITRO EFFECT OF GENOTYPE, GROWTH SEASON

AND CYTOKININES ON PEACH VARIETIES
(Prunus persica (L.) Batsch) PROPAGATION
Nazar AL GHASHEEM1, 2, Florin STĂNICĂ1, Adrian George PETICILĂ1, Oana VENAT1

1
University of Agronomic Sciences and Veterinary Medicine of Bucharest, 59 Marasti Blvd,
2
College of Agricultural and Marshes, Thi - Qar University, Iraq
Abstract
In vitro propagation is one of the most convenient methods for plant material multiplication in order to obtain virus
free planting material in high quantity and short time. The paper presents the influences of genotype, growth season
and hormones on in vitro propagation of some new Romanian peach varieties with very good prospects on the market.
Three peach genotypes: ʻFlorinʼ, ʻFilipʼ and ʻMimiʼ from the Didactic Field of Faculty of Horticulture, University of
Agronomic Sciences and Veterinary Medicine of Bucharest were tested. In the protocol for the in vitro establishment,
winter and spring growth season were chosen. Initial shoot explants were obtained in winter by forcing the dormant
shoots in the growing chamber and in spring (April-May), directly from the field. The explants were cultured on MS
(Murashige and Skoog, 1962) basal medium supplemented with 30 g sucrose and 7 g agar, as carbon source. Benzyl
aminopurine (BAP) on four variants with 0, 1, 5 and, respectively, 10 mg/l was used as cytokinine. The results show
that Florin genotype was superior to the rest of the varieties used in the experiment in terms of the number of formed
shoots and the strength of their growth. For the in vitro culture initiation, the shoots taken in spring, during the growth
season, gave the best results compared to the winter period. The results showed a significant correlation between the
concentration of BAP and the shoots number (multiplication rate) and height. The concentration of 5 mg/l BAP (V3),
gave the best rate of shoot formation and the highest elongation rate.
Keywords: benzyl aminopurine, culture media, shoots, multiplication rate, elongation rate
INTRODUCTION sophisticated laboratory procedure with unique

facilities and special skills (Hartmann et al.,
Peach (Prunus persica (L.) Batsch) is one of 2004; Sathyanarayana, 2007).
the most popular stone fruits. Peaches belong to Micropropagation is affected by many factors
Prunoideae, Prunus genus a subfamily of such as genotype, plant growth regulators
Rosaceae. It is one of the most popular (PGRs), agar, type of explants, culture medium
deciduous fruits and ranks second to apple and light conditions etc. For example,
among temperate zone deciduous fruit trees cytokinins are major factors in the induction of
from the standpoint of production and value somatic organs (George, 1993; Feyissa et al.,
(Childers, 1978; USDA, 2017). 2005).
China is the centre of origin for peaches and Several experiments were carried out for the
nectarines and was domesticated there 4000 multiplication of wood plants by tissue culture
years ago (Wang and Zhuang, 2001). Peach such as Ferradini N. et al. (1996) on apple
fruits are of high nutritional value because they rootstock and Peticila A.G. (2012) on kiwi. In
contain high levels of carbohydrates, fats, salts addition, the difficulty of regenerating plants
and vitamins and are used in the treatment of from mature tissues of woody plants is well
anaemia, poor digestion and nourishment of the established (Smigocki et al., 1991).
nervous systems (Al-Sheikh, 2003). Micropro- Peach is one of the most recalcitrant species
pagation is one form of tissue culture which with regard to micropropagation (Bhansali et
allows the production of large number of plants al., 1990; Padilla et al., 2006). Successful
from small pieces of the mother plant in regeneration of peach plants were from
relatively short period of time and limited immature seeds (Meng and Zhou., 1981;
space. It is an aseptic process which requires Hammerschlag et al., 1985; Scorza et al., 1990;
217
Bhansali et al., 1991; Smigocki et al., 1991; jars containing water to stimulate the sprouts to
Svircev et al., 1993; Pérez- Clemente et al., grow and break the dormancy (Figure 1). After
2004). Also regenerated from leaves explants two-three weeks were taken buds formed and
excised from shoots apexs culture (Gentile et used in the experiment, while the nodes were
al., 2002). used directly in the experiment. In the growing
Therefore, the main goal of this study was to season (spring: April-May) the explants were
establish a micropropagation protocol for taken and used directly in the experiment.
ʻFlorinʼ, ʻFilipʼ and ʻMimiʼ peach varieties in Media and culture conditions
order to produce a large scale of plants in a All explants were rinsed with ethanol 70% for
short period. Plant hormones are the most 2-3 min then were washed with distilled water
important effect factors in shoot regeneration three times for 2-3 min, Explants were surface
(Bhojwani and Razdan, 1996). Cytokinins are a sterilized with NaOCl (10% v/v) for 10-15 min.
type of plant growth regulators (PGRs). Then explants were washed with sterile
Contributed to in many processes of plants distilled water at least three times for 5 min.
growth, like cells division, Shoot and root MS (Moorashige and Skoog, 1962) consisting
morphogenesis. PGRs are regulating axillary of 30 g/l sucrose and 7 g/l agar without any
bud growth. Considering the importance of hormone supplements during initiation stage
these PGRs, among growth regulators used in and added 0, 1, 5, respectively, 10 mg/l Benzyl
peach tissue culture media are the cytokinins: aminopurine (BAP) during multiplication stage.
TDZ, kinetin and BAP, for auxins: IBA, IAA The pH of medium was adjusted to 5.6 with
and NAA (Hammerschlag, 1985; Mante et al., HCl 0.1 N or NaOH 0.1 N before sterilization
1989). This study aimed to evaluate different by autoclaving at 121°C for 15 minutes.
concentrations of BAP cytokinins for in vitro Sterilized explants were inoculated on culture
shoot development in peach. media and then placed in an incubation room at
22±2°C, and 16 hours daily (Stănică et al.,
MATERIALS AND METHODS 2002).
Statistical analysis
The study was conducted at the tissue culture The experiment was repeated two times, each
laboratory of the Faculty of Horticulture, treatment contained 10 replicates initiation
University of Agronomic Sciences and stage and 5 replicates tested on multiplication
Veterinary Medicine of Bucharest during the stage. All experiments were arranged in a
period October 2016 - June 2017 on Peach completely randomized design (CRD). Culture
(Prunus persica L.). Three peach varieties period ranged between four to eight weeks
(ʻFlorinʼ, ʻFilipʼ and ʻMimiʼ) were included in depending on individual experiment. Data were
the experiment. Explants were taken from trees recorded on shoots number formed, shoots
planted in the field of experiments to the length and leaves number were analyzed.
Faculty of Horticulture, University of Significance of differences between the results
Agricultural Sciences and Veterinary Medicine was estimated by Analysis of Variance
in Bucharest. Two explants types namely (ANOVA) on SPSS version 14 (SPSS 2005)
shoot-tips and nodes (one node with a single program with the means compared with LSD
axillary buds) were taken at 0.5-1cm length, test at < 0.05.
were tested on their ability to maintain and
initiate shoots on MS medium without any
hormone supplements during initiation stage
and added 0, 1, 5, respectively, 10 mg/l Benzyl
aminopurine (BAP) during multiplication stage.
V1=0 mg/l (control); V2=1 mg/l; V3=5 mg/l;
V4=10 mg/l.
Shoot-tips and nodes were obtained from two
sources (last year's growths, modern growths).
The explants were taken in the winter season
(dormancy buds) 15-20 cm long and placed in Figure 1. Peach branches placed in water
for break dormancy
218
RESULTS AND DISCUSSIONS hormone-free medium. It can be assumed that
the differences in their response in tissue
Analysis of variance (Tables 1 and 2) revealed culture were determined by the balance of their
that the treatment had highly significant effect endogenous hormones (Razdan, 2003). The
on mean leaves number and shoots length. The difference might be due to intra-metabolism of
results of means mean leaves number and plant which affected cell division and
shoots length formed developing in response to differentiation (Techato et al., 2002).
all the varieties of the experiment for the tissue The results showed that there were statistical
culture and there were significant differences differences in the shoots length of and leaves
between the varieties, where Florin variety had number formed by shoot-tips and nodes.
the largest number of leaves formed 5.00 leaf Results showed superiority shoot-tips on nodes
per explants in winter and 5.80 leaf per (Tables 1 and 2). Results showed that each type
explants in spring while the ʻFillipʼ and ʻMimiʼ of explants are characterized by a certain
varieties gave the least number of leaves. Also regeneration potential, depending on the
the ʻFlorinʼ variety gave the longest shoots species of plant and its degree of maturity,
formed (2.37 cm in winter and 4.77 cm in which is of physiological state of explants.
spring) while the ʻFilipʼ variety gave the Several types of explants have been widely
shortest shoots formed (1.69 cm in winter and used for in vitro such as Citrus lemon (Rathore
2.77 cm in spring). et al., 2004); young leaves on French bean
(Kamal and Praven, 1991); terminal buds of
renewal on gladioli (Rumynin et al., 1990);
apical buds on hybrid of mountain ash
(Suvorova et al., 1990) and Rough lemon (Ali
and Mizra, 2006).
Analysis of the results showed that there were
significant differences between leaves number
formed and shoots length from explants which
were taken from different seasons (Table. 2).
Explants taken in the growth season (spring)
gave the best results, as opposed to explants
which were taken in the winter for all varieties
(Table.1).
Previous studies have confirmed that there is a
relationship between phenolic compounds and
the age of the plant used, a common problem
reported in tissue culture of woody species (Mc
Figure 2. Hoot-tips of peach on MS Cown, 2000; Mathur et al., 1999). Ozyigit
after 2 week from culture
(2008) indicated a positive direct relationship
between age of explants and phenolic
There were no significant differences in shoot
exudation in tissue culture of cotton.
number per explants in initiation stage. There
Different combinations of cytokinins (BAP)
was a single shoot in all explants (Figure 2).
interacted significantly in terms of the shoots
Growth of roots in the hormone-free medium
number (Table 3). Variant V3 (BAP 5 mg/l)
was not observed. In multiplication stage also
gave the maximum shoots number (ʻFlorinʼ
there were significant differences between the
8.00, ʻFilipʼ 7.40 and ʻMimiʼ 5.40). Lowest
varieties. ʻFlorinʼ variety superior on ʻFilipʼ
number of shoot (1.00) were obtained in
and ʻMimiʼ varieties in the number and length
control medium (without any plant growth
of shoots formed (Figure 3). The effect of
regulators). Increasing BAP doses in
genotype on successful tissue culture has been
combination had an increasing effect up to a
previously reported (Gubis et al., 2003;
certain level V4 (BAP10 mg/l). Data presented
Blinstrubiene et al., 2004). Cotton callus
in higher shoots length were obtained in control
initiation (Zouzou et al., 1997; Zouzou et al.,
medium (without any plant growth regulators).
2000) at all the genotypes were cultured onto
219
This data shows that the shoot’s lengths were that the inclusion of BAP in shoot proliferation
markedly affected by various combinations of media enhanced the growth of axial shoots and
cytokinins. Statistically, after treatment V4 promoted the multiplication of shoots from the
(BAP 10 mg/l) with elevated levels of basal tissues of explants (Ohki and Sawaki,
cytokinins, shoot length decreased (ʻFlorinʼ 1999). A decline in the number of shoots with
2.59 cm, ʻFilipʼ 1.77 cm and ʻMimiʼ 1.94 cm). higher BAP levels has also been reported.
Analysis of LSD values between varieties and Waseem et al. (2009) showed that the use of
variants showed that the effect of BAP on the higher concentrations of PGRs may result in
shoots number and shoots lengths were plant weakness and decreased growth
significant at p≤0.05 (Tables 4 and 5). (Panjaitan et al., 2007).
Similar regeneration behaviours of BAP in pear Al-Sulaiman and Barakat (2010) cytokinins had
(Kadata and Numi, 2003); in peach rootstock a positive effect on the production of lateral
GF 677 (Ahmad et al., 2003); in bananas by shoots of Ziziphus spinachristi. The appropriate
(Vuylsteke, 1989; Arinaitwe et al., 2000). addition of cytokinin promotes the growth of
Previous researchers Vuylsteke and De Langhe shoots and reduces the dominance of the apical
(1985); Bairu et al. (2008) indicated that 5 mg/l bud (Asaad et al., 2009). In the Prunus species
BAP was the best concentration for banana the type of citokinine and its concentration are
varieties. important factors for multiplication and
In vitro multiplication rate was largely elongation rate (Leontiev-Orlov et al., 2000a).
controlled by interaction the varieties and Increasing (BAP) causing a rise in the numbers
cytokinins concentration and BAP is the most of buds primordia in chrysanthemum (Karim et
economical cytokinins (Gaspar et al., 1996; al., 2002, 2003). PGRs for bud break and shoot
Augusto,. 2001; Silveira et al., 2009). Rapid differentiation due to their role in cell
growth and multiplication of shoots are based multiplication and the breakdown of apical
on the quantity and quality of cytokinins and dominance (Casimiro et al., 2001). In woody
auxins in media as well as on their endogenous plants, BAP is paramount for growth compared
levels in plants. Histological studies showed to kinetin (Fráguas et al., 2004).
A B C
Figure 3. Peach genotypes after 3 weeks on V3 (MS+ BAP 5 mg/l): ʻFlorinʼ (A); ʻFillipʼ (B); ʻMimiʼ (C)
220
Table 1. Analysis of Means and Std. Deviation for the effect of genotype, growth season and explants on leaves number
and shoots length after 4 weeks cultivation on three peach varieties (ʻFlorinʼ, ʻFilipʼ and ʻMimiʼ)
Genotype Season Explants Mean leaves Std. Mean shoots Std.

number deviation length deviation
Florin Winter Shoot-tips 5.0000 1.00000 2.3740 .80640
Nodes 3.0000 1.22474 2.0180 .90502
Total 4.0000 1.49071 2.1960 .82961
Spring Shoot-tips 5.8000 1.09545 4.7760 .92802
Nodes 8.0000 2.34521 3.0220 .31260
Total 6.9000 2.07900 3.8990 1.13172
Total Shoot-tips 5.4000 1.07497 3.5750 1.50813
Nodes 5.5000 3.17105 2.5200 .82914
Total 5.4500 2.30503 3.0475 1.30227
Filip Winter Shoot-tips 3.8000 .83666 1.6900 .48974
Nodes 3.2000 1.30384 1.3960 .20671
Total 3.5000 1.08012 1.5430 .38678
Spring Shoot-tips 4.4000 .54772 2.7700 .53810
Nodes 4.6000 .54772 2.2960 .54344
Total 4.5000 .52705 2.5330 .56776
Total Shoot-tips 4.1000 .73786 2.2300 .74786
Nodes 3.9000 1.19722 1.8460 .61258
Total 4.0000 .97333 2.0380 .69389
Mimi Winter Shoot-tips 2.6000 .54772 1.6140 .68090
Nodes 2.4000 1.14018 1.2900 .29589
Total 2.5000 .84984 1.4520 .52357
Spring Shoot-tips 3.8000 1.30384 2.9420 .40752
Nodes 2.6000 .54772 1.9780 .13554
Total 3.2000 1.13529 2.4600 .58319
Total Shoot-tips 3.2000 1.13529 2.2780 .87735
Nodes 2.5000 .84984 1.6340 .42256
Total 2.8500 1.03999 1.9560 .74722
Total Winter Shoot-tips 3.8000 1.26491 1.8927 .71550
Nodes 2.8667 1.18723 1.5680 .61784
Total 3.3333 1.29544 1.7303 .67726
Spring Shoot-tips 4.6667 1.29099 3.4960 1.12216
Nodes 5.0667 2.65832 2.4320 .56753
Total 4.8667 2.06336 2.9640 1.02771
Total Shoot-tips 4.2333 1.33089 2.6943 1.23283
Nodes 3.9667 2.31164 2.0000 .72996
Total 4.1000 1.87490 2.3472 1.06373
221
Table.2. Analysis of variance (ANOVA) for the effect of genotype, growth season and explants on leaves number and
shoots length after 4 weeks cultivation on three peach varieties (ʻFlorinʼ, ʻFilipʼ and ʻMimiʼ)
Source Dependent Variable Sum of Squares df Mean Square F Sig.
Corrected Model Leaves number 144.200a 11 13.109 9.956 .000
Shoots length cm 50.635b 11 4.603 13.702 .000

Intercept Leaves number 1008.600 1 1008.600 766.025 .000
Shoots length cm 330.551 1 330.551 983.935 .000
Varieties Leaves number 67.900 2 33.950 25.785 .000
Shoots length cm 14.781 2 7.391 21.999 .000
Season Leaves number 35.267 1 35.267 26.785 .000
Shoots length cm 22.829 1 22.829 67.954 .000
Explants Leaves number 1.067 1 1.067 .810 .373
Shoots length cm 7.231 1 7.231 21.526 .000
Genotype *Season Leaves number 17.433 2 8.717 6.620 .003

*Explants
Shoots length cm .946 2 .473 1.408 .255
Genotype *Season Leaves number 14.233 2 7.117 5.405 .008
Shoots length cm 1.653 2 .826 2.460 .096
Genotype *Explants Leaves number 1.633 2 .817 .620 .542
Shoots length cm 1.145 2 .572 1.704 .193
Season *Explants Leaves number 6.667 1 6.667 5.063 .029
Shoots length cm 2.050 1 2.050 6.102 .017
Error Leaves number 63.200 48 1.317
Shoots length cm 16.126 48 .336
Total Leaves number 1216.000 60
Shoots length cm 397.312 60
Corrected Total Leaves number 207.400 59
Shoots length cm 66.760 59
222
Table. 3. Combined effect of genotype and concentrations of BAP on shoots number and shoots length per explants
Mean shoots length Std.

Genotypes Variants Mean shoots number Std. deviation
(cm) deviation
Florin V1 1.0000 .00000 5.7380 .67887
V2 4.0000 .70711 3.8440 .69540
V3 8.0000 1.87083 4.7760 .92802
V4 3.8000 1.30384 2.5920 .91319
Total 4.2000 2.78341 4.2375 1.40635
Filip V1 1.0000 .00000 3.3340 .69346
V2 2.4000 .54772 2.7700 .53810
V3 7.4000 1.51658 3.1440 .62616
V4 1.6000 .89443 1.7680 .43275
Total 3.1000 2.73188 2.7540 .81770
Mimi V1 1.0000 .00000 4.0980 .43390
V2 2.2000 .83666 2.9420 .40752
V3 5.4000 1.14018 3.8880 .44263
V4 1.2000 .44721 1.9400 .69199
Total 2.4500 1.93241 3.2170 .99420
Total V1 1.0000 .00000 4.3900 1.18345
V2 2.8667 1.06010 3.1853 .71137
V3 6.9333 1.83095 3.9360 .94388
V4 2.2000 1.47358 2.1000 .75069
Total 3.2500 2.57514 3.4028 1.24945
223
Table 4. LSD values between genotype on shoots number and shoots length per explants at P ≤ 0.05
(I) (J) Mean difference (I- Std. Error Sig. Mean difference Std. Sig.
Genotypes Varieties J) Shoots number (I-J) Shoots length Error
(cm)
Florin Filip 1.1000* .30687 .001 1.4835* .20450 .000

* *
Mimi 1.7500 .30687 .000 1.0205 .20450 .000
Filip Florin -1.1000* .30687 .001 -1.4835* .20450 .000
Mimi .6500* .30687 .039 -.4630* .20450 .028

*
Mimi Florin -1.7500 .30687 .000 -1.0205* .20450 .000
Filip -.6500* .30687 .039 .4630* .20450 .028
Based on observed means. The error term is mean square (error)=0.942. Shoots number. The error term is mean square
(error)=0.418. Shoots lengths (cm). *The mean difference is significant at the 0.05 level.
Table 5. LSD values between variants on shoots number and shoots length per explants at P ≤ 0.05
(I) Variants (J) Mean difference (I-J) Std. Sig. Mean difference Std. Sig.
Variants Shoots number Error (I-J) Shoots length Error
(cm)
*
V1 V2 -1.8667 .35434 .000 1.2047* .23614 .000
*
V3 -5.9333 .35434 .000 .4540 .23614 .060
V4 -1.2000* .35434 .001 2.2900* .23614 .000
* *
V2 V1 1.8667 .35434 .000 -1.2047 .23614 .000
V3 -4.0667* .35434 .000 -.7507* .23614 .003
V4 .6667 .35434 .066 1.0853* .23614 .000
*
V3 V1 5.9333 .35434 .000 -.4540 .23614 .060
V2 4.0667* .35434 .000 .7507* .23614 .003
V4 4.7333* .35434 .000 1.8360* .23614 .000
* *
V4 V1 1.2000 .35434 .001 -2.2900 .23614 .000
V2 -.6667 .35434 .066 -1.0853* .23614 .000
V3 -4.7333* .35434 .000 -1.8360* .23614 .000
Based on observed means. The error term is mean square (error)=0.942. Shoots number. The error term is mean square
(error)=0.418. Shoots lengths (cm). *The mean difference is significant at the 0.05 level.
CONCLUSIONS elongation rate was obtained at a concentration

of 5 mg/l BAP (V3).
The regenerative activity in the studied three
varieties depends on the highest level from the ACKNOWLEDGEMENTS
genotype, type of the explants and growing
season were better regeneration activity is The authors thank all colleagues, students,
registered with shoot-tip and spring season. The researchers, management and staff in
results showed a significant correlation micropropagation laboratory, Centre for
between the concentration of BAP and the Studies of Food and Agricultural Products
shoots number (multiplication rate) and height. Quality, USAMV for their assistance in this
The best rate of shoot formation and the highest work.
224
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226
IN VITRO EFFECT OF VARIOUS STERILIZATION TECHNIQUES

ON PEACH (Prunus persica (L.) Batsch) EXPLANTS
Nazar AL GHASHEEM1, 2, Florin STĂNICĂ1, Adrian George PETICILĂ1, Oana VENAT1

1
59 Marasti Blvd, District 1, Bucharest, Romania
2
College of Agricultural and Marshes, Thi- Qar University, Iraq
Corresponding author,e-mail:[email protected]
Abstract
One of the major challenges in Romania in fruit growing sector is the production of certified planting material,
considering the specificities of the local climate conditions and the consumers’ preferences. Due to the damages
produced by hard winters and late spring frosts and the lack of resistant and suitable peach varieties, in the last
decades, this species was almost eliminated from the producers choices in establishing new orchards. One of the major
research projects of the Faculty of Horticulture within the University of Agronomic Sciences and Veterinary Medicine
of Bucharest is to identify and multiply the best peach varieties (Prunus persica L. Batsch) adapted to the Romanian
harsh conditions. This paper presents different sterilization techniques applied to peach explants necessary for the
initiation of the in vitro culture. The research was conducted at the Micropropagation Laboratory within the Faculty of
Horticulture. For peach tissue culture initiation, four sterilization agents where tested in 18 different variants: Sodium
hypochlorite (NaOCl) in three concentrations: 5%, 10% and 15%, for 5 and 10 min; Hydrogen peroxide (H2O2), in two
concentrations: 5% and 10%, for 10 and 20 min; Captan (50%) fungicide, in four concentrations: 1%, 2%, 3% and 4%,
for 5 min and Boric acid (B(OH)3), in two concentrations: 1% and 2%, for 5 and 10 min. The explants (shoots-tip and
nodes) were cultured in MS (Murashige and Skoog, 1962) basal medium supplemented with 30g sucrose, as carbon
source and 7g agar. The growth chamber for the in vitro cultures had 22±2°C temperature and 70 to 80% relative
humidity, with a photoperiod of 16 h day light and 8 h dark. The percentage of contamination, survival rate and shoot
growth were studied during the initiation phase. Among the different sterilization variants, sodium hypochlorite was the
most effective treatment with 50% survival rate at V5 (15% for 5 min) and 60% at V4 (10% for 10 min). After
sterilization, shoots continued to grow vigorously and the multiplication phase initiated.
Key words: shoot-tip, node, sodium hypochlorite, hydrogen peroxide, boric acid.
INTRODUCTION in micropropagation, plant tissues on their

surfaces inherently have various bacteria and
Peach (Prunus persica L. Batsch) is one of the fungi, moulds etc. It is necessary that the
most important stone fruits which are grown explants be free from any surface contaminants
extensively in different parts of the world. prior to tissue culture since contaminants can
Peaches are native to China and their planting grow in the culture medium, rendering the
dates refer to at least 4000 years (Wang and culture non activity (Hiremath, 2006). There is
Zhuang, 2001). Peach trees are infected by so much pathogen (microbial contaminants)
numerous different pests and diseases. which has been a major threat to tissue cultures
Micropropagation which encompasses cell due to their rapid proliferation characteristics,
culture, tissue culture, organ and embryo microbes can be come from explants,
culture, has been a vital technique for laboratory instruments, conditions in the
considerable multiplication of plants, laboratory and contaminants may be introduced
elimination of plant diseases through tissue with the staff during manipulations in the
culture technique, plant conservation and laboratory (Leifert and Cassells, 2001; Enjalric
improvement production through gene transfer et al., 1998).
(Sarasan et al., 2011). Contamination is a real problem that opposes
Tissue culture techniques are used for the progress and development of tissue culture
commercial and research purposes extensively technology (Webster et al., 2003). These
to grow many different plants (Hussain et al, microbes compete adversely with explants for
2012). Aseptic conditions are usually practiced nutrients, and their presence often results in
227
variable growth or increased culture mortality seed and explants sterilization in many plants.
or can also result, reduced shoot proliferation, Hypochlorite was used to surface sterilize
tissue necrosis and reduced rooting (Oyebanji wheat seeds (Sauer and Burroughs, 1986).
et al., 2009). A successful in vitro culture Also sodium hypochlorite has been reported to
protocol, starts with effective explants be very effective factors different types of
sterilization, the sterilization chosen for an bacterial strains (Nakagawara et al., 1998).
experiment depend on the type of explants, Hydrogen peroxide solution as sterilizing agent
Sterilize material and plant genotype (Dodds has been reported for plants (Dumroese et al.,
and Roberts, 1985; Rezadost et al., 2013). 1988; Ogawa and Masaki, 2001; Rosner et al.,
There are several various sterilization factors 2003). It is recommended to use in the initial
are used to sterilize tissues, these disinfect sterilization as ethanol (Stănică et al., 2002).
materials are also toxic to explants tissues, and Hydrogen peroxide solution as sterilizing factor
therefore select the correct concentration and and improved germination on many plants like
the times of exposure to explants, must be wax, cotton, barley, pines, safflower and
elected to reduce the injury of plants (CPRI, currant (Dumroese et al., 1988; Rosner et al.,
1992). 2003; Cram and Fraedrich, 2009; Çavusoglu
So there is a state of balance between and Kabar, 2010; Lizarraga-Paulin et al., 2013).
sterilizing explants and killing the explants A report confirmed by Dolatabadian and
themselves (Qin et al., 2012; Olew et al., 2014). Modarressanavy (2008) hydrogen peroxide is
Many researchers have used these sterilizing more effective than other sterilization factors
agents successfully; also there are studies on for plant tissue. However, there are several
the effect of fungicides and antibiotics on these studies indicates that hydrogen peroxide was
kinds of contaminants (George, 1993; Rashid et ineffective for surface sterilization of explants
al., 2008; Maqbool et al., 2010; Bakhsh et al., and seeds (Miche and Balandreau, 2001).
2012). Several different mechanism are used to Captan 50% WP, agricultural fungicide for the
eliminate fungal and bacterial contamination, control of certain fungus diseases of fruit,
including the use of inactivation by heat and vegetables and ornamental crops are used in
light, fungicides and antibiotics, the time of surface sterilization in vitro.
sterilization is dependent on the type of tissue There are several studies like reported by
(Haldeman et al., 1987; Kneifel and Leonhardt, Sohnle et al. (1998). Also the effectiveness of
1992; Leifert et al., 1992). Bavistin was confirmed by Garla et al. (2011).
Explants are commonly surface-sterilized using Reported by Altan, et al. (2010), it failed to
ethanol, sodium hypochlorite, mercuric inhibit the microorganism and the activity of
chloride, hydrogen peroxide, fungicides and these sterilizing factors. Shields et al. (1984)
antibiotics. Therefore, the present study was analyzed the effects of fungicides against in
conducted to compare different sterilizing vitro on tobacco cultures. They recommend two
protocols for peach micropropagation and to fungicides, carbendazim and fenbendazole.
find out the best, efficient and cost effective Boric acid in agriculture is used as an
sterilization procedure that may result in least insecticide, herbicide and fungicide in food
or no contamination in peach tissue culture. In crops and orchards (EPA. U.S., 1993). Used in
our study we compared several modifications the United States as a fungicide on citrus
of four surface sterilization methods based on (Olkowski et al. 1993). The results by Jan H. et
the use of, sodium hypochlorite, hydrogen al. (2002) indicate that use of boric acid (3%
peroxide, captan (50%) and boric acid with solution) as pre-plant seed treatment on potato
using explants of peach accessions with was gave the lowest percent incidence of
different degrees of contamination. powdery scab diseases 4.3%.
However, in this study we did not used the
mercuric chloride, this material is very MATERIALS AND METHODS
dangerous because of more difficult to dispose
and high toxicity (Li et al., 2005; Jean-Philippe Explants surface sterilization
et al., 2012). Sodium hypochlorite (bleach) is Shoot tips and nodes (0.5-1 cm in size) of
the most common sterilization agent used for peach (Prunus persica L. Batsch cv ʻFlorinʼ)
228
were collected from the orchard of trees planted % of explants survived; mean length of shoots
in the field of experiments to the Faculty of (cm); mean number of leaves per explant.
Horticulture, University of Agronomic Experiments were conducted as factorial
Sciences and Veterinary Medicine of Bucharest experiments based on Completely Randomized
to used as explants for in vitro culture Design (CRD).
establishment.
Explants were placed under running tap water RESULTS AND DISCUSSIONS
with detergent for 30 min to remove any
foreign contaminants. Effect of sterilization factors
After washing, explants were dissected and The study showed there is an effect of
surface sterilized in a laminar air flow hood substances used in sterilization, sodium
with rinsed with ethanol 70% for 2-3 min then hypochlorite the most effective treatment with
were washed with distilled water three times 50% survival rate in 15% for 5 min and 60% in
for 2-3 min after that. For peach tissue culture 10% for 10 min and has outperformed the rest
initiation, four sterilization agents where tested of the other sterilizers which their results were
in 18 different variants: Sodium hypochlorite not satisfactory as the results were hydrogen
(NaOCl) in three concentrations: 5%, 10% and peroxide (H2O2) with 25% survival rate in 10%
15%, for 5 and 10 min (Figures 3, 4); Hydrogen for 20 min; Captan 50% with 25% survival rate
peroxide (H2O2), in two concentrations: 5% and in 4% for 5 min; Boric acid (B(OH)3) with 20%
10%, for 10 and 20 min (Figure 1); Captan survival rate in 2% for 10 min (Table 2).
(50%) WP fungicide, in four concentrations: These results are similar to the studies Satish et
1%, 2%, 3% and 4%, for 5 min (Figure 2) and al. (2012) on sugarcane; Siddique et al. (2018)
Boric acid (B(OH)3) in two concentrations: 1% on Skimmia laureola, when they used different
and 2%, for 5 and 10 min. (Table 1). substances in sterilization, where the results
differed according to the sterilizers.
Culture media and culture conditions Studied by many researchers, the solution of
The explants (shoots-tips and nodes) were sodium hypochlorite for superficial sterilization
cultured in MS (Murashige and Skoog, 1962) of explant was efficient and didn't injury the
basal medium supplemented with 30 g sucrose, explants at appropriate focus (Gertlowski K.
as carbon source and 7 g agar. pH was adjusted and Petersen M., 1993).
between 5.7 and 5.8 by using either 1 N HCl or These results are similar with those of
1 N NaOH before the agar was added. Hippolyte (2000), which reference that the high
Media was then heated on a hot plate with focus of sodium hypochlorite can be effective
continuous stirring using a magnetic stirrer in sterilizing the superficial explants cultivated
until agar is dissolved and media put in the in vitro, but it is accompanied by the death of
culture tubes. The culture tubes were covered explants.
with lids and put in trays and autoclaved. Many researchers have found these sterilizing
Autoclave was adjusted at a temperature of agents successfully (Rashid et al., 2008;
121°C for 15 min. The growth chamber for the Maqbool et al., 2010; Bakhsh et al., 2012).
in vitro cultures had 22±2°C temperature and
80-85% relative humidity, with a photoperiod Effect of concentration and exposure time
of 16 h day light and 8h dark. (Stănică et al., The study showed that there was a correlation
2002) between the dipping period and the concen-
tration of the substance used in the sterilization
Data collection on the extent of their effect on the percentage
In the experiment ten replicates (one explants of explants survived, explants contaminated by
in one tube culture) were used for each fungi, bacteria and explants dead due to the
treatment and the experiment was repeated increased concentration of the material used
twice. (Tables 2, 3 and 4).
Results were taken after 2, 4 weeks of planting Increasing the exposure duration and sterile
and the following data were recorded; % of concentration had reduced the contamination
contamination (fungus+ bacteria and sterilizer);
229
rate but highest number of loss explants Effect of explants
resulted. The study showed there are differences in the
The influence of sterilizing chemical ruin the extent of the response of the explants used in
shape and functions of microbe's enzymes rate of plant growth (shoots length and leaves
(George et al., 2008). number), contaminated rate and survival rate
But the increasing exposure duration and (Tables 3 and 4). Also, the shoots had been
concentration of sterilizes above certain registered the lowest rate of infection and the
optimum limit cause loss of explants because of most effect to increase the concentration of
the oxidant chemical ingredient ruin the plant sterile material compared to the contract also
tissue as well (Danso et al., 2011). explained that the shoots and nodes gave the
These findings are similar of the negative best rate of shoots length and leaves number
effects of Sodium hypochloride at high formed when using concentrations less (Figures
concentration were observed (Colgecen et al., 3 and 4).
2011). These results are similar with (Rezadost et al,
And a higher concentration of hydrogen 2013) who confirmed that the surface sterilize
peroxide 5% was reported to negatively affect used for an experiment typically depend on the
in sunflower (Dolatabadian and explants and plant species.
Modarressanavy, 2008).
Figure 3. Peach explants (shoots-tip) after 28 day from

Figure 1. Fungal contamination on peach explants after sterilization V10 ( NaOCl 10% for 10 min). There are
14 day from sterilization V9 (H2O2 10% for 10 min) leaves damaged because of the sterilizer
Figure 2. Fungal contamination on peach explants after

14 day from sterilization V11 (captan 1% for 5 min)
Figure 4. Peach explants (nodes) after 28 day from

sterilization V10 (NaOCl 10% for 10 min)
230
Table 1. Types of sterilizing agents used in a different concentration with varying time of sterilizings on peach explants
Pre-sterilization Exposure
Concentration Exposure time Surface Concentration
Variants Substance time
% (min) sterilizer %
disinfectants (min)
V1 Ethanol 70% 2-3 Sodium hypochlorite 5 5
V7 Ethanol 70% 2-3 Hydrogen peroxide 5 10
V11 Ethanol 70% 2-3 Captan 50% 1 5
V15 Ethanol 70% 2-3 Boric acid 1 5
Table 2. Effect of surface sterilizer, various concentrations and time exposure on % of contamination and % of survived
on explants after 14, 28 days from culture
Exposure
Surface Concentration
Variants time Shoot-tips Nodes
sterilizer %
(min)
Contamination Survived Contamination Survived
% % % %
after after after after
14 28 14 28
days days days days
V1 Sodium hypochlorite 5 5 70 100 00 80 100 00
V7 Hydrogen peroxide 5 10 70 85 15 70 85 15
V11 Captan 50% 1 5 35 90 10 50 100 00
V12 Captan 50% 2 5 35 90 10 70 100 00
V13 Captan 50% 3 5 30 80 20 40 85 15
V14 Captan 50% 4 5 30 75 25 45 85 15
V15 Boric acid 1 5 50 100 00 70 100 00
V16 Boric acid 1 10 50 85 15 60 100 00
V17 Boric acid 2 5 40 80 20 70 90 10
V18 Boric acid 2 10 45 80 20 80 85 15
231
Table 3. Effect of surface sterilizer, various concentrations and time exposure on % of contamination
(fungus + bacteria and sterilizers) on explants after 28 days from culture
Exposure
sterilizer %
(min)
Contamination %
Contamination %
after 28 fungus+ after 28 fungus+
sterilizer sterilizer
days bacteria days bacteria
V3 Sodium hypochlorite 10 5 70 59.5 10.5 70 63 7
V5 Sodium hypochlorite 15 5 50 15 35 50 42.50 7.50
V10 Hydrogen peroxide 10 20 75 52.5 22.5 80 68 12
V11 Captan 50% 1 5 90 90 00 100 100 00
V12 Captan 50% 2 5 90 90 00 100 100 00
V13 Captan 50% 3 5 80 56.25 23.75 85 76.50 8.50
V14 Captan 50% 4 5 75 56.25 18.75 85 67.25 12.75
V15 Boric acid 1 5 100 100 00 100 100 00
V16 Boric acid 1 10 85 85 00 100 100 00
V17 Boric acid 2 5 80 60 20 90 65.50 4.50
V18 Boric acid 2 10 80 60 20 85 67.25 12.75
Table 4. Effect of surface sterilizer, various concentrations and time exposure on mean shoots length (cm)
and mean (no) leaves shoot on explants after 28 days from culture
Exposure
sterilizer %
(min)
Mean Mean Mean Mean
Survived Survived
shoot shoot shoots shoot
after after
length Leaves length Leaves
28% 28%
(cm) (on) (cm) (on)
V1 Sodium hypochlorite 5 5 00 0.00 0.00 00 0.00 0.00
V7 Hydrogen peroxide 5 10 15 4.34 4.44 10 3.45 7.33
V11 Captan 50% 1 5 10 4.50 5.99 00 0.00 0.00
V12 Captan 50% 2 5 10 4.22 5.78 00 3.13 7.23
V13 Captan 50% 3 5 20 3.80 4.43 15 2.98 7.11
V14 Captan 50% 4 5 25 3.62 3.68 15 2.77 6.78
V15 Boric acid 1 5 00 0.00 0.00 00 0.00 0.00
V16 Boric acid 1 10 15 4.17 4.16 00 0.00 0.00
V17 Boric acid 2 5 20 3.87 4.20 10 3.01 6.89
V18 Boric acid 2 10 20 3.45 3.80 15 2.48 6.22
232
CONCLUSIONS decontamination and subsequent plantlet regeneration
of sugarcane Sacchrum officinarum L. in vitro. Int. J.
Integr. Biol. 11: 90-96.
Among the different sterilization protocols
Dolatabadian A., Modarressanavy S.A.M., 2008.
tested for the successful establishment of in Effect of ascorbic acid, pyridoxine and hydrogen
vitro culture of peach tissue culture. Our results peroxide treatments on germination, catalase activity,
showed that during the sterilization were protein and malondialdehyde content of three oil
different depend on the sterilization factors, seeds. Not. Bot. Horti. Agrobot. Cluj-Napoca. 36: 61-
66.
exposure time and explants type was used for
Dodds J.H., Roberts L.W., 1985. Experiments in Plant
micropropagation. Tissue Culture 2nd Ed. Cambridge University Press,
It is recommended for this study to be used Cambridge.
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sodium hypochlorite was the most effective 1988. Douglas-fir seed treatments: effects on seed
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(ed) Western Forest Nursery Council, Intermountain
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Pesticide Programs. Washington D.C. USA.
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234
SENSORY ANALYSIS OF THE DEHYDRATED PRODUCT

OBTAINED FROM APPLES HARVESTED FROM THE TRADITIONAL
ORCHARDS OF THE BRAN - ZĂRNEȘTI AREA
Cătălina ITIGAN, Florin STĂNICĂ

59 Marasti Blvd, District 1, Bucharest, Romania Romania
Abstract
Having a scientific foundation of more than two decades, sensory analysis is an easy method for assessing the quality of
a food product by bringing its characteristic features to the public consumers’ attention. As a complementary method in
food control and expertise, sensory analysis mirrors the outcome of certain physiological and psychological aspects of
the tasters, which involve such actions as reception, recognition, ordering, description, and appreciation. Relying on
the consumer’s first contact with the product, the analysis measures its sensory properties that affect the subjects’
choice power as well as their purchase decision. By describing the results obtained from the sensory analysis of apple
chips, the present paper seeks to emphasize the importance of tasting in the presentation, refinement and promotion of
the product. This objective was accomplished at the INDAGRA International Fair, which took place at Romexpo
Bucharest on 4th November 2016. The apple chips and other food products were featured by the stand of the University
of Agronomic Sciences and Veterinary Medicine of Bucharest. 191 people, 21-60 years old, were invited to visit the
stand of the University of Agronomic Sciences and Veterinary Medicine Bucharest and taste the products. The apple
chips were evaluated for different organoleptic features like appearance, taste and aroma as well as for the general
impression to simulate a possible authentication of the product as a preferred registered trademark. At the same time,
the experiment aimed to emphasize the importance of nutraceutical fruit consumption by attracting the consumer’s
attention with the present analysis and through raising awareness of healthy food and eating. The paper presents the
results, being an important step to follow with the serial production of the tested products.
Key words: apple chips, taste, appearance, aroma.
INTRODUCTION tasting in the presentation, refinement and

promotion of the product.
Having a scientific foundation of more than This objective was accomplished at the
two decades, sensory analysis is an easy INDAGRA International Fair, which took place
method for assessing the quality of a food at Romexpo Bucharest on 4th November 2016.
product by bringing its characteristic features to The apple chips and other food products were
the public consumers’ attention. featured by the stand of the University of
As a complementary method in food control Agronomic Sciences and Veterinary Medicine
and expertise, sensory analysis mirrors the of Bucharest.
outcome of certain physiological and To attain the objective, we planned on
psychological aspects of the tasters, which questioning the visitors open to taste the
involve such actions as reception, recognition, products and participate in the survey that
ordering, description, and appreciation. highlights the sensory properties of these
Relying on the consumer’s first contact with products.
the product, the analysis measures its sensory
properties that affect the subjects’ choice power MATERIALS AND METHODS
as well as their purchase decision.
By describing the results obtained from the Apples chips
sensory analysis of apple chips, the present Drying has been used in the United States for
paper seeks to emphasize the importance of more than a century (Downing, 1989) and
235
represents the process that underlies the The target consumers
obtaining of different food products like apple Intending to achieve a highly objective and
chips. Researcher S. Gould reported that at the complex result, the sensory analysis targeted
turn of the 19th century fruit dehydration had subjects of different age, gender and class.
been common for several years in the western Consequently, the realization of this selection
part of the New York state. was sought within the INDAGRA International
Shaped as rings or slices, apple chips are the Fair.
result of drying healthy, ripened fruit. 191 people, 21-60 years old, were invited to
Their thickness and shape are often irregular, visit the stand of the University of Agronomic
dehydration being a procedure that transforms Sciences and Veterinary Medicine of Bucharest
the fresh apple pieces in a particular way. and taste the products. Most of them, mainly
The drying proper implies the vaporization of males, gladly answered the invitation, some of
water within the fruit and its removal from the them showing an interest in forming or
used drying system. enriching their healthy diet culture.
As important as dehydration is product As expected, the subjects 40-60 years old were
packaging, which must ensure long-term present in the highest number, 70%, the
storage. younger people being responsive in a lower
At the same time, packaging must meet the percentage, 30%. As already mentioned, the
requirements of regular handling during male subjects outnumbered the female ones by
transportation and storage as well as of keeping 68.06%.
product integrity by removing moisture,
oxygen, light and foreign odour. Observance of Tasting the chips
packaging standards and norms guarantees In key with the particularity of the Bran -
product quality. Zărnești area, the apple chips obtained from the
Jonathan and Scortos cultivars were manually
The sensory analysis packed in small plastic bags decked with strips
As mentioned in the introduction, the quality featuring traditional folk themes. Likewise, the
measurement of the dehydrated products was products were exhibited in wattle baskets made
achieved through the sensory analysis method, by local craftsmen. The apple chips were
which recorded a wide range of customers’ offered in servings of four, as half-round slices.
preferences. The tasters were free to choose their preferred
The apple chips were evaluated for different products, which they did by picking one or
organoleptic features like appearance, taste and more food samples.
aroma as well as for the general impression to To measure and record the analysis results, the
simulate a possible authentication of the tasting was required to occur on the spot.
product as a preferred registered trademark.
At the same time, the experiment aimed to The tasting card
emphasize the importance of nutraceutical fruit The research method of the present experiment
consumption by attracting the consumer’s was the hedonic analysis, or the preference
attention with the present analysis and through test represented by the tasting card (Figure 1).
raising awareness of healthy food and eating.
236
Figure 1. Tasting card - research method
Figure 1 shows that the tasting card was Aiming at a highly objective result, the tasting
conceived by following certain parameters. By card did not require the subject’s name, except
putting a cross in a certain box, each taster their age and gender.
expressed their opinion about the appearance,
aroma, taste and general impression of the RESULTS AND DISCUSSIONS
product.
While the first three characteristics were The results were measured via a questionnaire,
associated with parameters like unpleasant, whose attitude scale interpreted the obtained
half-pleasant, neuter, pleasant, very pleasant, or information statistically.
no aroma, slightly aromatic, indifferent, The survey recorded relative and absolute
aromatic, very aromatic, the general impression frequencies, which were associated with
was measured in figures on a 1 to 5 scale. percentage and numerical results (Clinciu,
For a correct assessment, the figures were 2013).
associated with the following ratings: Accordingly, organoleptic dimensions like
unsatisfactory, satisfactory, good, very good aroma and taste recorded the results shown in
and excellent. the following tables (Tables 1, 2).
237
On the strength of Table 1, one can analyse the appreciated by the consumers and their market
tested consumers’ opinion about the aroma of production might be successful.
the prepared and presented chips.
It is noticeable that the female respondents Table 2. General taste determination
accept these chips more easily as they might Taste determination
become new and healthy prepared food for
both adults and children.
Unpleasant
Pleasant
pleasant
pleasant
Neuter
Respondents
Half-
Very
Table 1. Aroma determination
Indifferent
No aroma
Aromatic
aromatic
aromatic
Slightly
Respondents Male (%) - 0.76 1.54 40.78 56.92
Very
Female (%) - - 1.63 44.17 54.10
Male % 0.76 5.38 0.76 49.24 43.84
Female % - 1.63 - 47.54 50.82
In Figure 2, one can statistically notice that

females become more attracted by these natural
products as compared to males.
Figure 3. Chip taste determination by male and female

respondents
Except for one taster, who thought that the

product had no aroma, the others appreciated
Figure 2. Determination of apple chip aroma by the male
its quality. The vast majority considered that its
and female respondents
features were pleasant and very pleasant.
According to gender and age, the opinions
Chip taste determination leads to the varied from one subject to another, as the
observation that there is only a slight difference above tables clearly show.
between the male and female respondents. As a It was a welcome surprise to find that the
conclusion, the respondents’ taste does not vary appearance and general impression of the
too much. product recorded no parameters for unpleasant
Figure 3 is a graphical representation, and half-pleasant. Almost 100 percent of the
according to male and female respondents, of subjects opted for the pleasant and very
the individual taste of the analysed chips. pleasant appearance and general impression of
One can therefore say that both genders have the product.
close taste regarding the apple chips prepared It is worth remarking that more than 50% of the
in the laboratory. As more than 50% of the female subjects had a positive attitude,
respondents described the taste as being very classifying the product as very pleasant and
pleasant and more than 40% as pleasant, one excellent (Tables 3, 4).
can gather that these apple chips are
238
Table 3 recorded the data regarding the Table 4 statistically indicates that about 60% of
appearance of the chips obtained through the subjects are very impressed by the new
dehydration and drying. food products, while about 40% of them are
Following the data obtained from the examined only pleasantly impressed.
respondents, one can notice that the majority
are satisfied with the appearance of the Table 4. General impression
consumed chips, which are very similar to the
Unpleasant
classic chips such as those made from potatoes.
Pleasant
pleasant
pleasant
Neuter
Half-
Very
Respondents
Table 3. Appearance determination
Chip appearance determination 1 2 3 4 5
Male (%) - - 2,30 39.24 58.46
Unpleasan
Respondents Pleasant
pleasant
pleasant
Female (%) - - - 37.71 62.29
Neuter
Half-
Very
t
Figure 5 graphically demonstrates the same

Male (%) - - 3.08 43.84 53.08 tendency towards the appreciation of the
Female (%) - - - 42.62 57.38 remarkable quality of these Romanian apple
chips.
Statistically, as shown in the Figure 4 below,
both the male and female consumers share a
common opinion about the appearance of apple
chips.
Figure 4. Determination of the chips exterior appearance

by the male and female respondents Figure 5. Determination of the subjects’ general
impression regarding the chips obtained from apples
Feedback specific to the Bran - Zărnești area
The feedback obtained from the subjects of our
experiments shows that the females are more CONCLUSIONS
pleased with the new apple chips even as
compared to the ones made from potatoes, The Bran - Zărnești area is traditionally
while the males, at close quarters, are also appropriate for the growth of apple cultivars
pleasantly impressed by the new apple that have aromatic fruit and are resistant to the
products, which might be produced soon and cold climate of the region.
are healthy to boot. Likewise, the male and The aim of this research was to test the
female subjects appreciate all the quality consumer’s preferences on dried apples in
characteristics of these chips as well as the order to promote locally new possibilities of
aroma and the pleasant taste of the traditional valorisation of apple fruits.
Romanian apples which, compared to the The results showed that more than 62% of the
imported ones, are usually tasteless and have women and 58% of men appreciated this
no aroma, except for a pleasant appearance. product with the highest scores.
239
Both women and men evaluated the taste of the Romexpo INDAGRA International Fair, for
product with „pleasant” and „very pleasant” which we thank them.
scores, being a very important parameter of the This research activity was conducted with the
sensorial analysis. assistance of the University of Agronomic
For the dried apples it was chosen a package Sciences and Veterinary Medicine of
with national traditional aspects. The Bucharest.
consumers liked the theme, this parameter
being noted with the highest score also. REFERENCES
For the aroma parameter, the women showed
more interest than men, most of them Râpeanu G., 2010. Controlul falsificarilor produselor
appreciated it. alimentare. Tehnici si analize de laborator. Editura
Didactica si Pedagogica, Bucuresti.
The positive results of this study are an Cimpoieș Gh., 2012. Cultura marului. Editura Bons
important step for continuing the research in Offices, Chisinau.
the valorisation of the local products in the Heasman M., Mellentin J., 2001. The Functional foods
Bran - Zărnești area. revolution. Healthy people, healthy profits? Earthscan
Publications Ltd, London and Sterling, VA.
Braniște N., Uncheașu G., 2011. Determinator pentru
ACKNOWLEDGEMENTS soiuri de mere. Editura Ceres, Bucuresti, ISBN 978-
973-40-0401-5.
The present paper was done with the help of
the subjects who visited our stand at the
240
CUTTINGS PROPAGATION OF SOME FIG GENOTYPES (FICUS CARICA)
Holia AHMAD, Florin STĂNICĂ, Vlad STANCIU

Abstract
This paper presents the first result of the cuttings propagation of some genotypes of fig (Ficus carica) using an
integrated system of basal heating with artificial fog. Fig plants are spread in many regions of Romania, especially in
the south parts, and their fruits are very much appreciated by the consumers. Unfortunately, the offer of plant material
is very low on the market. In the research project of identification, monitoring and propagation of valuable fig
genotypes, more cuttings were collected from different genotypes and different regions of Romania and Iraq. Green and
dry cuttings were used and planted in benches for rooting with basal heating, in the greenhouses of the Research
Center for Studies of Food and Agricultural Products Quality at the UASVM Bucharest. Different substrates: sand +
perlite 70:30, sand + perlite +sawdust, perlite + peat + marc, wood chips with perlite, sand + perlite 60:40 were used.
The best results were obtained in the sand with perlite variants 70:30 (61.87%). After rooting, the new obtained plants
were transferred in pots and grown under controlled conditions (cold glass houses).
Key words: artificial mist, fig genotypes, rooted cuttings.
INTRODUCTION southern Banat, southern Oltenia and its Sub-

Carpathian depression, Dobrogea, as well as
Fig plants are cultivated on a large scale in around the main cities in the west and south of
tropical, subtropical and warm temperate areas the country. In these areas, local varieties or
of the world, especially in Morocco, Syria and varieties are grown, producing 1-2 crops per
Italy. Turkey is, however, the largest producer year depending on climatic conditions. In most
of dried figs (Sinha, 2003). crop areas, the fig plant is protected during
Despite its varietal diversity, fresh fig is still winter and, if planted in sheltered places, it can
considered as a minor fruit in trade with withstand even without protection.
Mediterranean countries (Mavsar et al., 2008). In plantations, plants need 4-5 m between rows
Fig plants are spread in many regions of and 2.5-4.0 m per row. In Romania it is grown
Romania, especially in the south parts, and in the garden next to the house, with promising
their fruits are very much appreciated by the results in winter protected areas (Stănică et al.,
consumers. Unfortunately, the offer of plant 2011).
material is very low on the market. Some of the A perennial woody plant, the fig plant behaves
major challenges for the fig plants are the like a shrub or even a tree in the conditions of
resistance to winter frost and the quality of Romania, and in the pedoclimatic conditions in
fruits. Being considered a species of interest for Iraq it can reach heights up to 12 m (Ghena et
Romania, many authors have presented over al., 2004). Due to the natural capacity of the
the years the biology and the peculiarities of species to form roots on sliced and rooted
the fig tree culture. portions of the plant, the fig plant can be
Cepoiu et al. (2005) mention in their book successfully multiplied by the vegetative
„Practical Fruit Growing” that in the climatic method by means of coventional methods, such
conditions of Romania, the fig plant can be as by dry or green cuttings (Stănică, 2002).
cultivated only protected over winter with Fig plant is multiplied vegetative by slips,
different materials or at the shelter of some cuttings or grafting. Because it has great
buildings. rooting capacity, dry cuttings is the most used
In „Pomology”, Hoza (2000) states that in our method in nurseries (Hoza, 2002; Grădinariu,
country the fig tree culture has developed in 2002)
241
This paper presents the first result of the 13 cm. The rooting bench in the dry place is
cuttings propagation of some genotypes of fig placed in a heated greenhouse, watering can be
(Ficus carica) using an integrated system of done manually or automated with a sprinkler
basal heating with artificial fog. system.
The rooting bank for green cuttings is made of
MATERIALS AND METHODS an aluminum vat about 20 centimeters deep
above which is mounted an arcade system that
In the research project of identification, supports a shading net that ensures future plants
monitoring and propagation of valuable fig maintain humidity and a temperature easier to
genotypes, more cuttings were collected from control. Also on the arcade system is mounted
different fig genotypes and different regions of the artificial fog system which is connected
Romania and Iraq. with a device called „artificial leaf” with the
Green and dry cuttings were used and planted purpose of starting and stopping the irrigation
in benches for rooting with basal heating, in the system. The biological material studied consists
greenhouses of the Research Center for Studies in different genotypes (Table 1).
of Food and Agricultural Products Quality at
the University of Agronomic Sciences and Table 1. Fig genotypes
Veterinary Medicine of Bucharest. Variant Genotype Origin
Different substrates: sand + perlite 70:30, sand V1 Secuilor 1 Bucharest, Romania
V2 Secuilor 2 Bucharest, Romania
+ perlite +sawdust, perlite + peat + marc, wood V3 Negoiești 01 Prahova, Romania
chips with perlite, sand + perlite 60:40 were V4 Str. Ștefan cel Mare Bucharest, Romania
used. After rooting, the new obtained plants V5 Muzeul Storck Bucharest, Romania
were transferred in pots and grown under V6 PH Corno Prahova, Romania
V9 Sebus Oradea, Bihor, Romania
controlled conditions (cold glass houses). In the V10 Stoica Dan Mangalia, Romania
same time, part of the collected biotypes are V11 Galben mare Giurgiu Giurgiu, Romania
planted in four testing plots at Pietroasa (Braniștea G.)
Viticulture and Wine Processing Research V17 Olimpia Tm Timișoara, Romania
V18 Smochin negru Ploiesti, Prahova, Romania
Station, Dăbuleni Research Station, Fruit V21 Str. Părintele Bucharest, Romania
Nursery and Farm Istrita in Buzau county and Stăniloae
Svinița Village, Mehedinți. All the plants are V28 Brazi Brazi, Prahova, România
monitored. V34 Rot negru Otiman Sviniţa, Caras-Severin,
Romania
Green cuttings: consists in cutting some pieces V42 Irak 1 Kerkuk, Irak
of seedlings of 13-15 cm, leaving the upper
leaves unsteady and the rest of the leaves out. RESULTS AND DISCUSSIONS
The cuttings thus obtained are inserted into the
rooting bed at a depth of 10-13 cm. They are The results obtained were synthesized in the
regularly wetted, requiring a humidity of 96- following tables and graphs.
98% and a temperature at the cutting level of Five months after planting in rooting lawns
20-22 degrees Celsius. (Figure 1), the cuttings were transplanted into
Cuttings are kept inside the rooting bench for pots and analyzed the following: the percentage
150-160 days after which they are carefully of viability of the plants obtained, the number
removed without breaking the roots. Depending of roots formed and their length, the size of the
on the degree of root development, the cuttings cuttings.
are planted in pots with a diameter of 15-20
centimeters.
Dry cuttings: it is harvested after the fall of the
leaves from November to the beginning of
March. An important aspect to be taken into
account when harvesting shoots during the cold
period is the degree of freezing during the
winter. The cuttings are shorten to 13-15 cm
and inserted into the substrate at a depth of 11- Figure 1. Fig plant cuttings detail
242
Table 2 presents the number of roots formed by From the measurements made it was found that
genotype and their size, 5 months after planting the substrate in which perlite and sand is in the
in the rooting bed (Figure 2). ratio of 30:70 proved to be the best substrate
for this type of propagation, confirming the
authors cited (Stănică et al, 2002; Hoza, 2000)
In order to observe the most effective method
of propagation, the percentage of those who
formed viable roots was transcribed when
rooted cuttings were planted in pots.
Thus, for the genotypes studied as a vegetative
propagation method, the dry one, which had a
rate of 42.07%, is favorable as opposed to the
one in the green which has a percentage of
14.69%. (Table 4)
Figure 2. Rooted cuttings detail Table 4. The degree of rooting according

to the type of cuttings
Table 2. Rooting results on fig genotypes Final
Initial Rooting
Cuttings rooted
Variant Lenght (cm) Roots (no) Average length (cm) No. cuttings percentage
type cuttings
(no) (%)
V5 40.33 7.33 5.23
(no)
1. Green 490 72 14.69
V9 86.67 9.67 8.57
2. Dry 920 387 42.07
V11 87.50 10.00 5.54
V17 35.40 5.80 6.04

Depending on the sun's exposure during the
V21 95.50 11.10 8.78 first two weeks of transplantation, the plants
V28 75.33 11.67 6.54 responded differently.
V34 84.67 9.00 9.77 They have undergone two different situations.
One of the situations requires the plant to
be exposed in full sunshine, and in the second
V21 genotype presented the best parameters
plants it is protected from the sun's rays
followed closed by V11, V9 and V34
(Table 5).
genotypes.
As a result of the vegetative propagation Table 5. The influence of shading on survival
activities of fig plants, the influence made by percentage of transplanted cuttings
the composition of the substrate in the rooting
process was made (Table 3). Viable
Cuttings
Growth cuttings after Rate
No. into pots
Table 3. Rooting percentage of cuttings depending conditions two weeks (%)
(no)
on the substrate used in the bench (no)
1. Sun 55 34 61.81%
Number of banks Percentage of
No. 2. Shadow 144 129 89.91%
(substrate type) success (%)
Bank I
1. 61.87
sand + perlite (70:30) As a result of the measurements made it was
Bank II found that the sun exposure of the plants
2. sand + perlite + 30.00 transplanted on the pots is not indicated, so that
sawdust
at this stage the fig plants have to be sheltered
Bank III from the direct sun rays in accordance with the
3. 21.76
perlite + peat + marc
results presented by Stănică et al., 2002.
Bank IV Two years later measurements were made on
4. 44.50
sand + perlite (60:40)
plant growths, foliar surfaces, and the number
of fruits obtained.
243
In Table 6 one can observe the measurements CONCLUSIONS
made on pot plants.
For the fig genotypes studied the best
Table 6. Measurements made of potted plants vegetative propagation method was dry cuttings
Length Shoots Leaves Fruit instead of green cuttings.
Variant
(cm) (no.) (no.) (no.) The substrate in which perlite and sand is in the
52.60 3.20 12.80 2.60 ratio of 30:70 proved to be the best substrate
V1
for this type of propagation. After transplanting
54.40 4.20 14.80 1.00
V2 the plants in pot, they have to be put in a
V3
63.00 1.62 14.00 4.25 shadow space for better results.
53.40 4.20 17.80 2.20
V21 genotype presented the best rooting
V4
parameters followed closed by V11, V9 and
V5 60.85 1.57 13.43 1.28 V34 genotypes.
71.87 1.50 10.37 2.50
V6
REFERENCES
V10 54.20 1.40 15.40 2.40
71.80 1.60 12.40 2.40 Ahmad H., Stănică F., Al Masoody M.M., Butcaru A.C.,
V11
2017. Preliminary Characterization of some Fig
V18
4.20 2.20 8.30 4.25 Biotypes in Romania, Bulletin USAMV,
Horticulture, vol. 74, 117-122.
46.25 2.00 11.50 1.75
V34 Cepoiu N., Păun C., Spița V., 2005. Pomicultură
21.75 1.50 8.00 1.25 practică. Editura Ceres, București.
V42
Ghena N., Braniște N., Stănică F., 2004. Pomicultura
generală. Editura Matrixrom, București.
It can be noticed that the largest length of the Grădinariu G., 2002. Pomicultura specială. Editura Ion
Ionescu de la Brad, Iași.
plants was made at genotypes V6 and V11 Hoza D., 2000. Pomologie. Editura „S.A.”, Prahova.
(71.8 cm). Mavsar D.B. editor, 2008. The Common Fig (Ficus
The largest number of shoots was formed at carica L.). Publishing House Annales Koper, Istrian.
genotypes V2 and V4 (4.2). The average Sinha K.K., 2003. Figs, T.M. Bhagalpur University,
number of leaves per plant had values between Bhagalpur, India.
Stănică F, Branişte N., 2011, Ghid pentru pomicultori.
8.0 for genotype V42 and 17.80 for genotype Editura Ceres, București.
V4. Most genotypes have fruits, but the average Stănică F., Dumitraşcu M., Davidescu V., Madjar R.,
value did not exceed 5 fruits per plant. The Peticilă A., 2002, Înmulţirea plantelor horticole
highest value was recorded in genotypes V3 lemnoase. Editura Ceres, București.
and V18 (4.25).
244
VITICULTURE
AND OENOLOGY
245
246
CHARACTERIZATION OF GRAPE AND WINE QUALITY INFLUENCED

BY TERROIR IN DIFFERENT ECOSYSTEMS FROM ROMANIA
CULTIVATED WITH FETEASCĂ NEAGRĂ
Alin DOBREI, Alina Georgeta DOBREI, Eleonora NISTOR,

Gheorghe POȘTA, Mihaela MĂLĂESCU, Marius BALINT
1
Banat University of Agricultural Sciences and Veterinary Medicine “King Michael I of Romania”,
Calea Aradului 119, Timișoara, Romania
Abstract
Grapes and especially wine from same variety may be very different, depending on many factors. These attributes have
the advantage that, different tastes, curiosities and demands of wine connoisseurs can be satisfied. The experiments
concerning the influence of terroir on the grapes and wine quality in ‘Fetească Neagră’ variety were located in the
main vineyards from western Romania (Minis-Maderat, Recas and Buzias-Silagiu). The aim of the research was to
found the influence of the ecological resources, type of pruning, fruit loading and age plantation on grapes quality
(sugar, acidity, must yield), but also on the wine quantity and quality (liters per hectare, alcohol concentration,
organoleptic characteristics, anthocyanin concentration, acidity, full-bodied). Although belongs to the same variety, the
resulted wines had different characteristics on most parameters analyzed. Recas wines are fruity; those from Buzias are
more obvious full bodied, while those from Minis-Maderat have higher anthocyanin pigmentation. After results
analyzing, it can be concluded that in all three areas of research, terroir allowed obtaining quality wines, but with
different characteristics, each bearing the mark of the origin area.
Key words: acidity, grapes, sugar, terroir, wine.
INTRODUCTION proper technology and winemaking, remarkable

results have been achieved (Dobrei et al.,
The soil, climatic and technological diversity in 2016b).
which the grapevine is cultivated have major A harmonious correlation of the crop
impact on wine quality - even in the same technology (fruit breeding, green pruning and
variety - so that wines with certain quality and training, optimum harvesting time), with the
typicality can be obtained, that can satisfy a ecological resources and climatic conditions of
wide range of consumers (Gladstones, 2011). the crop year, most often resulted in the
Quality cannot be achieved only through a ‘Fetească neagră’ grapes of high quality,
harmonious combination of factors: soil- without which it is not possible to obtain
climate-soil-technology (Dobrei et al., 2016a). reference wines (Belda et al., 2017; Cichi et al.,
In red wines, the influence of terroir on quality 2016).
is high, resulting in wines with different A harmonious correlation of the crop
properties (body and balance, anthocyanin technology (crop load, green pruning, favorable
pigmentation, tannins etc. - Dobrei A. et al., harvesting time) with the ecological resources
2010). of the area and with the climatic conditions,
Wine texture qualities are essential in have often resulted in obtaining grapes of
determining wine quality classes and are special quality in ‘Fetească neagră’ variety,
determined in accordance with wine tasting and without which it is not possible to obtain
typicity which reflects varietal origins (Dobrei special wines (Rotaru et al., 2010; Rotaru et al.,
et al., 2016a). 2013).
One of the most valuable red varieties is the
‘Fetească neagră’, considered by many MATERIALS AND METHODS
scientists and consumers, the most valuable
Romanian variety. However, although not Resarch was carried out during 2016-2017 in
highly advertised in last decades, through a four different locations: Buziaș-Silagiu, Recaș,
247
Miniș-Măderat and Mocrea vineyards. Mocrea relation equivalents / L sample) and was
was recognized from 1199 as wine-growing calculated according to the following relation:
settlement, now part of Miniș-Măderat
vineyards. (A MW DF 1000 )
Anthocyani ns (mg/l)
The complex influences of terroir - climate, L
soils, slope, and vineyard growing technology -
on grapes and wine quality from ‘Fetească where: A = (A520 nm pH 1.0 - A700 nm pH
neagră’ variety was investigated. 1.0) - (A520 pH 4.5 nm - A700 nm pH 4.5);
For more accurate results, the experiments were MW = molecular weight of cyanidin-3-
located in vineyards with different glucoside (449.2); DF = dilution factor; ε =
characteristics (vine age, vine training system, molar absorbance of cyanidin-3-glucoside in
crop load) placed on different soil types and acidic aqueous solution; L = optical path (1
land with various exposure to sunlight (Table cm).
1). Research years were climatically different: The method of wine colour determination with
2017 has been marked by high temperatures the highest accuracy was done by the
and low rainfall, while 2016 was a warmer year spectrophotometric method. This involves
and more rainfall, but without exceeding the raising VIS spectra (in the visible wavelength:
normal. 380-720 nm) in reflectance or transmittance.
Temperature associated with vineyard The sensory analysis was carried out in
placement and soil, has strong control on accordance with the Regulation of the
ripening and then on grape and wine quality. Association of Authorized Tasters in Romania
Located not far away from each other, all three and according to the International Vine and
vineyards produced wine for long time. Wine Office.
However, several geo-morphological variations According to the OIV (International Vine and
which induce different micro-climatic Wine Organization) quality assessment system,
conditions between vineyards affect the wine is assessed on three main characteristics
grapevine development and growth. and nine sub-characteristics, as well as a global
Grape maturity during ripening was determined assessment called harmony.
by sugar concentration and titratable acidity, in The main characteristics that were evaluated
the third decade of September in 2016 and are: appearance (turbidity or haze, color),
second decade of September in 2017. aroma, taste and mouthfeel (acidity, sweetness,
Grape samples were harvested from bitterness, astringency, any new flavors or
experimental plots from each vineyard in which aromas perceived). The tasting was done by a
the research was carried out. five member group, under laboratory
After berries crushing, the samples of must conditions, using tasting glasses, wines having
were subjected to analyze for sugars and total a temperature of 18°C.
acidity determination. The rating was divided into the following
Sugar concentration (g/l) was measured using a categories, based on the average of the points
digital refractometer at 20oC. received, obtained by adding the five notes and
The acidity of the samples was performed by dividing them to five.
the titrimetric method by neutralizing the The score obtained was as follows: excellent
acidity of the samples with a known 0.1 M (full body); very good (medium to full body);
NaOH solution (F = 0.9527). The obtained good (medium).
results are expressed in g/l tartaric acid. Results concerning the influence of terroir from
Winemaking technology has not been the experimental variants on ‘Fetească neagră’
subject of our research, this being carried by a grapes production and quality (sugar, acidity,
standard protocol specific to each vineyard. must yield), but also on the wine quantity and
Wine analyses were done after finishing the quality (wine yield per hectare, alcohol by
winemaking process. volume, wine sensory properties and texture,
Total content of monomeric anthocyanins anthocyanin pigmentation) were statistically
(cyanidin-3-glucoside equivalents / L sample) analyzed. All experiments were repeated three
was calculated according to the following times.
248
Table 1. Experimental variants
Exposure to Vine age Crop load
Vineyard Soil type Vine training system
sunlight (years) (buds/vine)
V1 - Recaș Reddish brown S-E 20 Guyot 20
Luvisols
V2 - Buziaș E 12 Cazenave 25
Podzoluvisols
V3 - Mocrea Rendzina soils S-E 6 Cazenave 16
V4 - Miniș Rendzina soils S 15 Guyot 18
Average and standard deviations of the data Buziaș and 10758 kg/ha at Recaș. In Miniș and
were calculated. The average comparison test Mocrea vineyards, yields were lower compared
and the Fisher test (LSD) were applied to to those from Buziaș and Recaș, but in normal
compare the results of must and wine limits for ‘Fetească neagră’ variety. In these
characteristics. The confidence interval was set areas, rendzina soils are considered to have
at 5%. For all statistical tests, the SPSS (SPSS limited potential for grapevine production.
Inc., Chicago, IL) for Windows, version 22.0 Concerning the grapes sugar content, results
was used. were good considering the climatic conditions
variation during the year; the highest sugar
RESULTS AND DISCUSSIONS content was recorded in Mocrea vineyard,
followed by results from Miniș and Recaș.
Year 2016 was normal for grapevine growing, Higher sugar concentrations recorded in the
without extremely favourable climatic Mocrea and Miniș areas are statistically
conditions due to the high rainfall in July, significant, as result of the rendzina soils
August and September. properties which lowered the crop load.
Grape yields were high, close to the maximum Regarding the must yield, differences between
crop potential, especially in the Buziaș and investigated variants are insignificant, with
Recaș vineyards (Table 2). In these areas the limits between 75-79%, values considered
soil fertility correlated with rainfall and crop normal.
load increased the production to 11375 kg/ha at
Table 2. The influence of terroir on production quality during 2016
Experimental Production Sugars Acidity Must yield Difference to control Significance
variant (kg/ha) (g/l must) (g/l H2SO4 ) (%) (sugars g/l )
V1 - Recaș 10758 209 4.9 79 -4 -
V2 - Buziaș 11375 198 5.2 78 -15 00
V3 - Mocrea 8773 226 4.6 75 +13 **
V4 - Miniș 9145 218 4.8 76 +5 *
Mean (Mt) 10013 213 4.8 77 - -
DL 5% - 4.93 1% - 10.17 0.1% - 18.32
During 2017 climate conditions were high and 8152 kg/ha at Mocrea. The sugar
temperatures and low rainfall, with positive concentration increased in grape berries from
influence on grape quality, but with lower all four vineyards experimental plots, ranging
yields than the previous year (Table 3). The from 249 grams per liter in Mocrea and 218
yields ranged between 10873 kg/ha at Buziaș grams per liter in Buziaș.
Table 3. The influence of terroir on production quality during 2017
V1 - Recaș 10120 228 4.7 78.0 -4 -
V2 - Buziaș 10873 218 5.0 76.0 -14 00
V3 - Mocrea 8152 249 4.4 73.0 +17 ***
V4 - Miniș 8207 233 4.6 74.0 +1 -
Mean (Mt) 9338 232 4.6 75.2 - -
DL 5% - 4.03 1% - 9.24 0.1% - 16.92
249
Table 4. The influence of terroir on production quality during 2016 - 2017
V1 - Recaș 10439 219 4.8 78.5 -4 -
V2 - Buziaș 11124 208 5.1 77.0 -15 00
V3 - Mocrea 8463 238 4.5 74.0 +15 **
V4 - Miniș 8676 226 4.7 75.0 +3 -
Mean (Mt) 9676 223 4.7 76.1 - -
DL 5% - 4.64 1% - 9.43 0.1% - 17.20
Due to the climate variability from both years, the accumulation of medium amounts of sugars
for the accuracy of the results is presented an in must (208 g/l), which are lower compared to
average of the two years of research in Table 4. the other three grape-growing areas. This value
Regarding the production, were recorded for the sugars is the only one with negative
average and high values for this variety in statistical significance compared to the average
correlation with climatic conditions, soil type results found during the research.
and crop loads. The smallest productions were During 2016, the wine yield per hectare varied
obtained on less fertile rendzina soils and on between 8 531 l/ha at Buziaș and 6 404 l/ha at
lower crop loads from Mocrea and Miniș Mocrea vineyards (Table 5). The more fertile
vineyards. The more fertile soils from Buziaș soils from Buziaș and Recaș favoured a higher
and Recaș, in correlation with higher crop amount of wine per hectare compared with less
loads, create a positive effect on the average fertile rendzina soils from Miniș and Mocrea.
production potential of this variety and higher In contrast, rendzina soil provided higher
yields were recorded. alcohol concentration and higher anthocyanin
With respect to grapes quality, all four grape- content compared to fertile soils.
growing areas have proven to be very In 2017, high temperatures and low rainfall
favourable to ‘Fetească neagră’ variety, sugar decreased the amount of wine yield per hectare
concentration recorded in grape berries being in all grape-growing areas investigated, but
high or very high. instead had a positive influence on alcohol
Rendzina soils correlated with the favourable concentration and anthocyanin content. This
exposure to sunlight and smaller crop loads, year, the smallest wine yields per hectare were
favoured the accumulation of very high produced on rendzina soils from Miniș (5 826 l/
amounts of sugar (238 g/l) in the grape must ha) and Mocrea (5 787 l/ha) respectively (Table
from Mocrea and 226 g/l in grape must from 6).
Miniș. Very good grape quality was also Concerning the sensory wine properties, in both
recorded in Recaș vineyards, where the years, wines were marked by their clarity,
reddish-brown soil associated with the southern colour and body, with a plus for those produced
exposure of the vine rows and a moderate crop in 2017 (Table 7). These wines, by proper
load on long spurs/canes, favoured the storing and vintage can become reference
accumulation of 219 g/l sugar. wines with real chances of assertion in the
In Buziaș vineyards, the luvisols in correlation future national or international wine contests.
with a higher crop load on short canes favoured
Table 5. The influence of terroir on wine quality during 2016

Experimental Wine yield Alcohol (%) Anthocyanin Sensory properties qualities
variant (l/ha) (mg/l) Limpidity Color Body
V1 - Recaș 8176 12.0 233.0 Clear Red ruby Medium to full
V2 - Buziaș 8531 11.5 218.0 Clear Red ruby Medium
V3 - Mocrea 6404 13.1 236.0 Limpid Red ruby Full
V4 - Miniș 6675 12.7 238.0 Limpid Dark red Full
Mean (Mt) 7446 12.3 231.2 - - -
250
Table 6. The influence of terroir on wine quality during 2017
Experimental Wine yield Alcohol (%) Anthocyanin Sensory properties qualities
variant (l/ha) (mg/l) Limpidity Color Body
V1 - Recaș 7590 13.2 243 Clear Red ruby Medium to full
V2 - Buziaș 7937 12.7 224 Clear Red ruby Medium
V3 - Mocrea 5787 14.5 245 Limpid Dark red Full
V4 - Miniș 5826 13.5 248 Limpid Dark red Full
Mean (Mt) 6785 13.4 240 - - -
Table 7. The influence of terroir on wine quality during 2016 -2017

Experimental Wine yield Alcohol (%) Anthocyanin
variant (l/ha) (mg/l)
V1 - Recaș 7883 12.6 238.0
V2 – Buziaș 8234 12.1 221.0
V3 - Mocrea 6096 13.8 240.5
V4 – Miniș 6251 13.1 243.0
During the research made by Artem et al. alcohol concentration (12.8 - 13.4%), higher
(2014) on four red wine varieties, they found in values for acidity (6.33 - 5.77 mg/l) and for
‘Fetească neagră’, highest values for acidity anthocyanins (409 - 531 mg/l).
(7.72 g/l), and for anthocyanins (325.92 mg/l). Wine components are much or less correlated
The alcohol concentration found in the same with each other. Alcohol concentration (%)
research (13.55%), was quite similar with the from ‘Fetească neagră’ during 2016-2017 is
average of ‘Fetească neagră’ from our highly and negatively influenced by the wine
investigation (13.4%). Mori et al. (2007) yield (l/ha) and by the crop load. The higher the
mention in their research that temperature over crop load, the lowest is the alcohol
350C decreased the anthocyanin content in red concentration in the wine. The wine yield is
grape berries. In a study carried out during strong influenced by the crop load. The
2014-2015, Coldea et al., found out in anthocyanin content in the wine is low
‘Fetească neagră’ variety a lower total acidity, correlated with crop load, while alcohol
between 3.28 and 3.72 mg/l, and alcohol concentration in the wine is negative correlated
content with limits between 11.78 and 13.70%. with the anthocyanin content and crop load
In the research carried out by Artem et al. (Table 8). Strong positive correlation was
(2015) in Murfatlar vineyards on ‘Fetească recorded between wine yield and crop load.
neagră’ variety, they found out close results for
Table 8. Correlation between wine yield, anthocyanin and crop load in ‘Fetească neagră’ variety, during 2016-2017
Wine yield Alcohol (%) Anthocyanin Crop load
(l/ha) (mg/l) (buds/vine)
Wine yield (l/ha) 7883 1 -0.936946137 0.545198709 0.989044201
Alcohol (%) 12.6 -0.936946137 1 -0.217855545 -0.978270352
Anthocyanin (mg/ l) 243 0.545198709 -0.217855545 1 0.415475039
Crop load (buds/vine) 20 0.989044201 -0.978270352 0.415475039 1
Table 9. Variability in Fetească neagră variety traits during 2016-2017, in research locations
Wine yield (l/ha) Alcohol (%) Anthocyanin (mg/l) Crop load (buds/vine)
Mean 6860.33 13.00 233.17 19.67
Standard Error 688.28 0.49 6.13 2.73
Standard Deviation 1192.15 0.85 10.61 4.73
Minimum 6096.00 12.10 221.00 16.00
Maximum 8234.00 13.80 240.50 25.00
CV % 17.37.00 6.57 4.55 24.05
251
As Jackson and Lombard (1993) said, the ensures customer demands through the special
quality of the wine “is not easy to define”, but sensory qualities, typicity and authenticity.
it is sure that a good wine must have good taste,
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254
STUDY OF THE ANTHOCYANIC POTENTIAL OF GRAPES VARIETIES

FOR RED WINES IN DRANIC WINE CENTER
Camelia MUNTEAN, Felicia STOICA, Constantin BĂDUCĂ CÂMPEANU,

Daniela Doloris CICHI
University of Craiova, Faculty of Horticulture, 13 A.I. Cuza Street, Craiova, Romania
Abstract
The Dranic plantation is one of the youngest in Dolj county and was founded in 2010 through the process of
reconversion and restructuring of the wine sector in our country. Under the favorable ecologically conditions for
viticulture in the Oltenia area, the varieties for red wines besides the superior potential of glucidic accumulation, have
a remarkable ability for biosynthesis of anthocyanins. The chromatic structures of the anthocyanin extracts represented
by the yellow, red and blue pigments are balanced, attractive and fully in line with current requirements. They are
dimensioned by colour intensity values, but especially by the qualitative chromatic indicators: the colour tone and the
proportions of the flavylium cations. This study may be the start for the suitability of this area for obtaining high-quality
red wines.
Key words: anthocyanin, grape maturity, red, wines, extractability, chromatic structure.
INTRODUCTION Anthocyanins are contained in the grape skin,

except in the case of a few cultivars whose pulp
The Dranic plantation is one of the youngest in is also pigmented. Thus, a close relationship
Dolj county and was founded in 2010 through can be expected between anthocyanin
the process of reconversion and restructuring of composition and visual appearance (Fernandez-
the wine sector in our country. In the absence Lopez et al., 1998). The International Office of
of clear data from the literature showing the the Vine plants (OIV) has a descriptor list for
direct relationship between natural factors and grape vine varieties and Vitis species which
the evolution of the red wine grape ripening classifies the grape varieties in seven groups
process, this detailed study was needed. This according to their external colour: 1 (green-
study may be the start for the suitability of this yellow), 2 (pink), 3 (red), 4 (red-grey), 5 (red-
area for obtaining high-quality red wines dark violet), 6 (blue-black) and 7 (red-black)
(Muntean et al., 2017). (OIV, 1988).
Phenolics, such as flavonoids, phenolic acids, Anthocyanins are gradually accumulated in
and tannins, are considered to be the main berry skins from veraison through grape
antioxidants in fruits and vegetables (Mnari et ripening (Gomez-Plaza, 2006; Fournand et al.,
al., 2016). 2006) malvidin-3-glucoside being the most
Phenolic compounds, such as phenols, phenolic abundant anthocyanin in almost all red grape
acids, flavonoids, tannins, and anthocyanins, varieties. However, the anthocyanin
have received considerable attention for their concentration may decline just before harvest
high antioxidant activity (Karakaya et al., 2001; and/or during over-ripening (Rolle et al., 2011).
Rice-Evans et al., 1996). Phenolic compounds The relationship between cell wall composition
are free radical scavengers because they are and extractability of anthocyanins from red
nucleophiles that inhibit lipid peroxidation and grape skins was assessed in Tempranillo grape
chelators of metal ions that induce oxidation samples harvested at three stages of ripening
(Han and Baik, 2008). (pre-harvest, harvest and over-ripening) and
Anthocyanins, the pigments responsible for the three different contents of soluble solids (22, 24
red colour of grapes, are widely used to carry and 26 °Brix) within each stage (Hernandez-
out classification and chemotaxonomic studies Hierro et al., 2014).
in flavonoid metabolism.
255
The extractability of anthocyanins from wine- These analyzes were conducted in 3
grapes with different skin hardness at two consecutive viticultural years, 2015, 2016 and
different ripening stages was evaluated by Role 2017.
et al (2009). Significant interactions between
ripening stage and skin hardness were found in Anthocynins extraction from grapes
the composition of individual anthocyanins Grape anthocyanins determined by the method
present in the extract. of Poissant Leon. An average sample of 50
Some researchers Romero-Cascales et al., in grape berries, weighed than the skin are
2005, were studying the differences in carefully detached from the grape pulp. In
anthocyanin extractability from grapes to wines order to remove excess moisture, the skins
according to variety. The anthocyanin were wiped with filter paper and dried in a hot
concentration of Vitis vinifera L. cvs. ʻCabernet air source.
Sauvignonʼ, ʻMerlotʼ, ʻSyrahʼ and ʻMonastrellʼ Dry skins are powdered quartz fine sand after
grape skins was determined together with which they are passed into a flask with ground
anthocyanin extractability at the exact time of glass stopper by repeated washing with 1% HCl
harvest (measured by an extractability assay solution (approx. 10 ml concentrated HCl/liter).
based on the comparison of the anthocyanin In order to extract anthocynins first wash
concentration of two different solutions fraction (50 ml) to be in contact with the skins
obtained after macerating the grapes for four to about 12 hours. The reafter, the filtered or
hours at two different pH values). These data centrifuged extraction liquid is brought to a
were compared with the anthocyanin constant volume of 200 ml by mixing and the
concentration and chromatic characteristics of fractions resulting from repeated acid addition
the resulting wines. and kept in contact with the extract for at least
The purpose of this study was to investigate the one hour.
evolution of anthocyanin content during grapes After obtaining the skin sample and the
maturation and the degree of extraction in anthocynins extract, the filtered extract is read
wines on a 1 cm cuvette spectrophotometer at the
optical density (OD) of 520 nm. The amount of
MATERIALS AND METHODS anthocynins is determined on the basis of the
formula:
Grape and wine sample mg kg grapes
OD 520 22 .76 0.4
1000
This study was carried out with grapes of the berries weight
three cultivars Vitis vinifera cv. ʻCabernet Anthocynins from wines
Sauvignonʼ, ʻMerlotʼ and ʻFetească neagrăʼ Anthocyanins in wine are determined with
from the wine-growing center Dranic-Dolj. spectrophotometer by pH difference (method
Grape samples of three red cultivars (V. Ribereau Gayon - Stonestreet - 1968). The
vinifera L.) were collected at full maturity difference between the optical densities read at
(FM) and at different other date after full 520 nm of a solution at two different pH is
maturity, means FM+10 days and FM+15 days. proportional to the amount of anthocynins
The grapes was harvest at FM+10 days which contains the wine. From the optical density of
corespond with technological maturity. the solution of pH 0.6, the optical density of the
The wine was obtained under the same solution of pH 3.5 is decreased, thus obtaining
biotechnological conditions of red vinification the PA. The concentration in anthocynins is
which was SO2 - 50 mg/l; ADY addition 15 determined from the calibration curve, based on
g/hl; 7 days maceration time; maceration the data, and the expression is expressed in
temperature 26-280C. mg/l.
Determination of anthocyanins from grapes and The colour of red wines (Glories, 1984) is
wine, pigments structure and flavilium cation determined with a T 70+UV/Vis spectrometer
from grapes and also chromatic composition PG Instruments Ltd. in the visible field at wave
and intensity and tonality of wine was done. lengths of 420, 520 and 620 nm.
After reading the yellow component at 420 nm
(attributed to tannins and anthocyanin
256
degradation products), the red component at Sauvignonʼ grapes is explained, as well as the
520 nm (attributed mainly to the free values of the parameter for brilliance for red is
anthocyanins as favylium cations and to the high (dA% - 76.5 - 79.3). The ʻFetească
polymeric red pigments) and the blue naegrăʼ variety showed the lowest anthocyanin
component at 620 nm (attributed to content and the highest yellow pigments (39.5-
anthocyanin quinoidal bases and 41.7), while the blue component showed the
copigmentation derived from a non-covalent lowest value (8.1-8.9%).
complex copigment - anthocyanin), the colour On the other hand Merlot variety, showed an
intensity (Ic), the colour tone (Tc) and the intermediate characteristics, achieving a
flavylium cation-brilliance of red (dA%) ratios balanced chromatic structure with positive
are calculated according to the following visual valences.
formulas: In the three years of study, the phenolic
maturity was performed after full maturity, at
Ic OD 420 OD520 OD 620 about 10 days, between 15.X. and 25.X. for the
ʻCabernet Sauvignonʼ variety, between 10.X.
OD 420 and 20.X. for the ʻMerlotʼ variety. The
Tc
OD 520 maximum amount of anthocyanins in grapes
does not correspond to the maximum amount of
OD 420 OD 620 1 anthocyanins in wines. Correlated with
dA% OD 520 OD 520 100
2 phenolic maturity, the phenomenon is
explained by the increase of extracts of
RESULTS AND DISCUSSIONS anthocyanins from the skins to the over
ripening of the grapes. At over-ripening,
The anthocyanic potential and the chromatic anthocyanin content in berries is lower, but
structure of the grapes of the main varieties for wine has higher anthocyanin content.
high quality red wines grown in the center of Table 2 shows the evolution of anthocyanin
the Winery are listed in Table 1. content in grapes and their degree of extraction
By its genetic nature, the histological structure at full maturity and over-ripening in 2017, a
of the berry and the ability to harness the year of excessively high temperatures.
natural conditions superior to the ʻCabernet In all 3 varieties observed, although total
Sauvignonʼ, accumulate the highest anthocyanin content decreases continuously, it
anthocyanin content, at the technological increases, in turn, its extraction due to the
maturity ranging from 1394 - 1493 mg/kg of degradation of the walls of the anthocyanin
fresh berries. The second place is occupied by blasts of the hypodermis, under the action of its
the Merlot variety, containing a concentration own pectolytic enzymes.
in anthocyanin ranging from 1205 to 1273 Thus, the proportion of extracted anthocyanins
mg/kg of fresh berries. Modest contents of increases from the 41.5% to 54.1% (ʻCabernet
anthocyanin accumulates in grapes of ʻFetească Sauvignonʼ), from the 41.7% to 52.5%
neagrăʼ variety being below 1200 mg/kg of (ʻMerlotʼ) and from the 40.8% to 53.5%
fresh berries. (ʻFetească neagrăʼ).
The chromatic structure, given by the In the wines obtained in 2017 the chromatic
proportions of the different categories of structure of the obtained wines has a different
pigments, varies from one species to another configuration (Table 3). As far as the over
and from one year to the next. Thus, in the ripening of the grapes - raw material, in the
anthocyanin complex of ʻCabernet Sauvignonʼ anthocyanic complex extracted under the action
grapes, the yellow-orange pigments have the of biotechnological factors, uniformly applied,
lowest values among the study varieties (31.5 - they increase the proportions of yellow and red
33.9%) while the blue pigments are the most pigments and decrease the proportions of blue
abundant (9.2 - 9.7%). The largest proportions pigments, which is still a qualitative advantage
of red and blue pigments have been of wine coloring, even when younger.
accumulated, and the intensity of the Values optical density at 420 nm wavelengths,
anthocyanins extracts from the ʻCabernet 520 nm and 620 nm specific to different types
257
of pigments used to calculate the qualities ʻFetească neagrăʼ. The blue component of
chromaticity wines, listed in Table 3 reveals wines is well and positively correlated with the
differences in some cases quite important. They red component.
are mostly of the genetic nature of variety and An accurate image of the quantity and quality
less of primary winemaking technology. of material coloured wine is obtained from:
Yellow component showed the lowest value in absolute content of anthocyanins, the
wines produced from ʻCabernet Sauvignonʼ, participation percentage of different types of
and the largest values in wines from ʻFetească pigments and values qualities chromaticity for
neagrăʼ while the red component showed the their definition being considered optical density
highest values in wines of ʻCabernet values (referred to in Table 4).
Sauvignonʼ and the lowest value in wines of
Table 1. Anthocyanin content and chromatic structure in grapes, technological maturity
(Average 2015-2017)
Antocyanin, Yellow pigments Red pigments Blue pigments Brilliance of red,

Varieties mg/kg fresh OD 420 nm OD 520 nm OD 620 nm dA %
berries
Variation Average Variation Average Variation Average Variation Average Variation Average
Cabernet 1394.0 - 1443.5 31.5- 32.7 56.8- 57.9 9.2-9.7 9.4 76.5- 77.9
Sauvignon 1493.0 33.9 59.0 79.3
Merlot 1205.0 - 1239.0 34.5- 35.1 54.6- 55.7 8.9-9.5 9.2 74.6- 75.0
1273.0 35.8 56.8 75.5
Fetească 1099.0 - 1119.5 39.5- 40.0 49.5- 51.4 8.1-8.9 8.5 72.5- 73.3
neagră 1140.0 41.7 53.4 74.2
Table 2. Anthocyanin content of grapes and their degree extraction, depending on the harvesting period – 2017
STAGE OF MATURITY
Full maturity (FM) FM + 10 days FM + 15 days
Varieties Total Extractable Degree of Total Extractable Degree Total Extractable Degree of
anthocyanin, anthocyanin, extr., anthocyanin, anthocyanin, of extr., anthocyanin, anthocyanin, extr.,
mg/kg of mg/kg of % mg/kg of mg/kg of % mg/kg of mg/kg of %
fresh berries fresh berries fresh berries fresh berries fresh berries fresh berries
Cabernet
1498.0 623.2 41.5 1493.0 744.5 49.8 1469.0 795.4 54.1
Sauvignon
Merlot 1281.0 534.7 41.7 1273.0 598.0 46.9 1192.0 625.8 52.5
Fetească
1146.0 468.5 40.8 1140.0 584.6 51.2 1132.0 605.8 53.5
neagră
Table 3. The optical densities of red wines varieties 2017
OD 420 nm OD 520 nm OD 620 nm

Varieties
(yellow pigments) (red pigments) (blue pigments)
Cabernet Sauvignon 0.489 0.832 0.140
Merlot 0.492 0.737 0.125
Fetească neagră 0.522 0.658 0.115
Table 4. Chromatic composition of red wines 2017
Composition of pigments % Intensity Tonality of

Varieties Yellow pigm. Red pigm. Blue pigm. of colour colour
(OD 420 nm) (OD 520 nm) (OD 620 nm) (Ic) (Tc)
Cabernet Sauvignon 33.35 57.05 9.60 1.46 0.58
Merlot 36.50 54.30 9.20 1.35 0.67
Fetească neagră 40.30 50.80 8.90 1.29 0.79
Data on participation of various types of levels of characteristics chromatic of complex
pigment in composition of coloring matters and anthocyanins, show structures chromatic very
258
favorable to all wines and proportions of Han H., Baik B.K., 2008. Antioxidant Activity and
different types of pigments are able to provide Phenolic Content of Lentils (Lens culinaris),
Chickpeas (Cicer arietinum L.), Peas (Pisum sativum
levels of color corresponding total over the L.) and Soybeans (Glycine max), and Their
claims current. Quantitative Changes During Processing.
International Journal of Food Science and
CONCLUSIONS Technology, 43, 1971-1978.
Hernandez-Hierro J.M, Leticia Martínez Lapuente, Belén
Ayestarán, Rivas-Gonzalo J.C., Teresa Escribano-
The natural conditions that are particularly Bailón, 2014. Relationship between skin cell wall
favorable to the vineyards in the Dranic composition and anthocyanin extractability of Vitis
vineyard and significantly mark the vinifera L. cv. ʻTempranilloʼ at different grape
anthocyanin - rich content of the grapes of the ripeness degree, Food Chemistry, Vol. 146, 41-47.
ʻCabernet Sauvignonʼ, ʻMerlotʼ and even of the Karakaya S., El S., Tas A., 2001. Antioxidant Activity of
Some Foods Containing Phenolic Compounds.
native ʻFetească neagrăʼ variety. International Journal of Food Sciences and Nutrition
During the over-ripening process of grapes - a 2001, 52, 501-508.
condition that is generally required for the Mnari A.B., Arij Harzallah A., Amri Z, Aguir S.D.,
production of high class red wines, anthocyanin Hammami M., 2016. Phytochemical Content,
content decreases with the progress of process, Antioxidant Properties, and Phenolic Profile of
Tunisian Raisin Varieties (Vitis vinifera L.),
but in the same time it was observed an International Journal of Food Properties, 19: 578-590.
increase of the extraction degree and, on the Muntean Camelia, Stoica Felicia, Baduca Campeanu C.
other hand, chromatic characteristics of the Rotaru Iulia, 2017. Relationship between the main
resulted wines are improved substantially. quality parameters and antocyanic content during
grapes ripening in Dranic viticultural area, Annals of
the University of Craiova - Agriculture,
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Fernandez-Lopez J.A., Almela L., Munoz J.A., Hidalgo OIV, 1988. Nouvelles varieties de raisins de table et de
V., Carreno J., 1998. Dependence between colour and raisins secs. Ed. OIV, Paris.
individual anthocyanin content in ripening grapes, Rice-Evans C.A., Miller J.M., Paganga G., 1996.
Food Research International, vol. 31, no. 9, p. 667- Structure-Antioxidant Activity Relationship of
672. Flavonoids and Phenolic Acids. Free Radical Biology
Fournand D., Vicens A., Sidhoum L., Souquet J.M., Medical 1996, 20, 933-956.
Moutounet M., Cheynier V., 2006. Accumulation and Rolle L., Río Segade S., Torchio F., Giacosa S.,
extractability of grape skin tannins and anthocyanins Cagnasso E., Marengo F., Gerbi V., 2011. Influence
at different advanced physiological stages. J. Agric. of Grape Density and Harvest Date on Changes in
Food Chem. 2006, 54, 7331-7338. Phenolic Composition, Phenol Extractability Indices,
Gomez-Plaza E., 2006. The effect of grape ripening stage Journal of Agricultural and Food Chemistry, 59,
on red wine color. J. Int. Sci. Vigne Vin 2006, 40, 15- 8796-8805.
24.
259
260
THE BEHAVIOR OF THREE CABERNET SAUVIGNON CLONES

IN VALEA CĂLUGĂREASCĂ AREA
Vlad-Andrei FILIP1, Elena BRÎNDUȘE1, Marian ION1, Petruța Mihaela MATEI2,

Laura Lidia CĂLUGĂRU3
1
Research and Development Institute for Viticulture and Oenology, Valea Călugărească,
Mantei Street no. 1, 107620, Prahova, Romania
2
District 1, 011464, Bucharest, Romania
3
National Office of Vine and Wine Products, 49 Iancului Street, District 2, 021719,
Bucharest, Romania
Abstract
Cabernet Sauvignon variety is one of the most widespread grapevine for wine grown in Romania. By application of
clonal selection, the only method to avoid the degeneration phenomenon of varieties to biotypes, five clones of Cabernet
Sauvignon have been obtained and homologated in Romania. The present study aimed to analyse the behaviour of three
Cabernet Sauvignon clones, namely Cabernet Sauvignon 30 Vl., 131 St. and 54 Mn. in the Valea Calugareasca
viticultural area, considering the extension of their culture. The experiments were carried out during 2016-2017 harvest
seasons, and were referred to the phenotypic spectrum, the fertility and productivity characteristics, the biological
potential, the maturation dynamics of the grapes, the qualitative and quantitative evaluation of the grapes at harvest.
These three Cabernet Sauvignon clones showed a similar phenological cycle and the behaviour to biotic factors,
resistance to the pathogen attack being good and very good. During ripening, the sugar accumulation was similar at all
clones (between 2.38 and 2.63 g/day) the amount of sugar accumulated on maturation period being between 49.9g and
55.2g. The glucoacidimetric ratio recorded very high values, characteristic for exceptional years (over 60%), mainly
due to the very low value of total acidity. Acids metabolism has been achieved on average with 1.59-1.73 g/day. In
terms of grape weight, the clone Cabernet Sauvignon 30 Vl. was highlighted with a greater weight of bunches with
11.5% compared with the Cabernet Sauvignon 54 Mn. and with 12.41% compared with the Cabernet Sauvignon 131 St.
From the point of view of the polyphenolic composition, Cabernet Sauvignon 54 Mn. was noted by the value of total
polyphenol index and the anthocyanic potential.
Key words: biotypes, clonal selection, Cabernet Sauvignon, glucoacidimetric ratio, polyphenol index.
INTRODUCTION better quality of wines obtained (Stefanini et

al., 2000; Tebeica et al., 2005). From a great
Cabernet Sauvignon is one of the most number of Cabernet Sauvignon clones, wines
widespread vinegrapes varieties for wine grown with distinctive flavour of fruit aroma, higher
in Romania (Șerdinescu et al., 2006; Urucu, content of tannins, anthocyanins etc. are
2014). By application of clonal selection, the produced in France (Catalogue, 2009), Italy
only method to avoid the degeneration (Fidelibus et al., 2006), Australia and other
phenomenon of varieties to biotypes, five countries.
clones of Cabernet Sauvignon have been The present study aimed to analyse the
obtained and homologated in Romania. Many behaviour of three Cabernet Sauvignon clones,
authors have reported the distinction of some in the Valea Calugareasca viticultural area,
Cabernet Sauvignon clones from the point of namely Cabernet Sauvignon 30 Vl. obtained by
view of wines quality, distinctive flavour of ICDVV (Research-Development Institute for
fruit aroma, higher content of tannins, Viticulture and Oenology, Valea Calugareasca)
anthocyanins (Jones and Davis, 2000; Fidelibus and approved in 2010, Cabernet Sauvignon 131
et al., 2006; Dejeu, 1986). St., obtained by INCDBH (National Research
Clones of one variety differ from the and Development Institute for Biotechnology in
population in better features of the grape and Horticulture, Stefanesti Arges) and approved in
261
2000 and Cabernet Sauvignon 54 Mn. obtained Absolute and relative fertility coefficients were
by SCDVV (Research and Development calculated using computation formulas based
Station for Viticulture and Oenology, Minis on the number of inflorescences, total shoots
Arad) and approved in 2006, considering the and fertile shoots. The productivity of varieties
extension of their culture. was determined at full grape maturation, using
absolute and relative productivity indices and
MATERIALS AND METHODS the weight of the grapes (Pop, 2003). The
average weight of grape at full maturity was
Three Cabernet Sauvignon clones, namely calculated by weighting 50 grapes per clones.
Cabernet Sauvignon 30 Vl., Cabernet The sugar concentration was determined by a
Sauvignon 131 St., and Cabernet Sauvignon 54 hand held digital refractometer and the results
Mn., were taken into study. were expressed as an absolute value and as a
The vines, were grafted on the SO4 percentage by mass of sucrose (OIV-MA-AS2-
(Oppenheim Selection 4) rootstock, were 02 method). Titratable acidity was determined
planted in 2014 in the germplasm collection by titration with 0.1M NaOH, with 1%
belonging to the Research and Development phenolphthalein and the results were expressed
Institute for Viticulture and Oenology, Valea in gL–1 H2SO4 (OIV-MA-AS313-01 method).
Calugareasca. 24 vines per genotype were The behaviour to biotic factors was assessed by
taken into study. The evaluation of clonal using OIV ampelographic descriptor method,
accessions focused on the duration of their the notation being done through attribution of
phenological cycles, grape fertility and figures depending on the level of expression.
productivity, resistance to diseases, quantity The phenolic potential was assessed by the
and quality of the grapes production. standard ITV (Institute Technique de la Vigne-
The recording of the vegetation phenophases France) method (www.vignevin-sudouest.com)
was carried out weekly, following the based on the following analytical parameters:
methodology defined by the OIV (International anthocyanins, total anthocyanin potential and
Organization of Vine and Wine) descriptor list total polyphenol index.
for grape varieties and Vitis species (2009). Extraction of phenolic compounds from the
Time of bud burst was noticed when 50% of grapes was done with an aqueous acid solution.
the buds are in green shoot tip stage C of Maceration of the samples was done for one
Baggiolini, stage 7 to 9 of BBCH (Biologische, hour at room temperature. Fifteen milliliters of
Bundesanstalt, Bundessortenamt und ethanol (95%) and 85 mL of 0.1% HCl were
CHemische Industrie scale). Time of full bloom added to the 50 g of grape juice. The samples
was noticed when 50% of flowers are open. were shacked for one minute from quarter to
When 50% of the red grape clusters show quarter. Coloured extracts are filtered through
changes in colour, or when about 50% of the glasswool (or quantitative filter paper) in order
berries of the white cluster start softening was to obtain clear solutions.
noticed the time of beginning of berry ripening The dilution of the samples to 1/100 was done
(veraison). Time of physiological stage of full in double-distilled water followed by the
maturity of the berry is related to the maximum measurement of the absorbance at 280 nm in a
sugar content of the berry due to 1 cm quartz vat, nm against a blank of distilled
photosynthesis. Mean value of all bunches of water and the total polyphenolic index (IPT)
10 shoots was taken into account. was determined as follow:
The behaviour to biotic factors was assessed by Total polyphenolic index (IPT) = DO 280 x
using OIV ampelographic descriptor method, 100 x [(weight of marc + 100)/ weight of
the notation being done through attribution of marc].
figures depending on the level of expression. The samples were diluted 1/20 in 1%
Production and the quality of grape harvest was hydrochloric acid solution and absorbance was
calculated based on fertility coefficient and measured at DO 520 nm against a blank of
productivity index, the average length and distilled water. The concentration of
average weight of bunch grapes and berries, anthocyanin and the total anthocyanic potential
sugar content and the total acidity of must. were determined following formulas:
262
Anthocyanins (mg/l) = DO 520 x 22.75 x 20 indicates the presence of a relatively warm and
Total anthocyanic potential (mg/kg) = drier climate. The active heat balance of the
anthocyanins (mg/l) x [(weight of marc + 100)/ vegetation period records an average value of
weight of marc]. 3379°C and the thermal balance is 1779°C.
The precipitation from the vegetation period
RESULTS AND DISCUSSIONS amounted to an average of 411.3 mm.
The high heliothermic regime in July, August
Compared to multiannual averages, the wine and September, on a normal rainfall regime,
year 2016-2017 is characterized by a higher even low in August and September, caused a
thermal regime, due to a relatively normal good accumulation of sugars in grapes (Matei
rainfall regime. During the winter, there were et al., 2009).
no minimum temperatures to affect the viability
of the fruit buds, with an absolute minimum of The development of the vegetation
-12.8°C, recorded in January, compared to - phenophases
13.6°C multiannual average. Related to the climatic conditions of 2016 year,
Both medium and absolute highs temperatures in Valea Calugareasca viticultural centre, it has
were (with a few exceptions) much higher been noticed that all the clones entered in
compared to multi-year averages. It should be vegetation at the end of March.
noted the high temperature regime in the winter The beginning of budburst occurred on 30th
months, when active temperature values of March at Cabernet Sauvignon 30 Vl. and two
11.6°C were recorded in December (compared days later in case of the other two clones. This
to 5.7°C multiannual average) and 15.3°C in phenophase ended in the second decade of the
February 2016 (versus 14.7°C multiannual month of April, after sixteen days at Cabernet
average). Sauvignon 30 Vl. and after fourteen days at the
The precipitation rate was very low in the other two.
winter months, when it was 0.2 mm in The two days difference was also observed in
December compared to 44.5 mm in the the case of blossoming, 50% of the
multiannual average, but it became surplus in inflorescences at Cabernet Sauvignon 30 Vl.
the spring months (87.6 mm versus 35.3 mm being blossomed on 7th June, respectively on
multiannual average recorded in March) and in 9th June in case of the other two clones.
July, favouring the development of The ripening of the grapes started from 15th
cryptogamic diseases. In the warmest months August. 50% of the grape clusters showed
(June, July and August) there were 81.8 mm, changes in colour on 21st August, date when
70.2 mm and 78.2 mm. was noticed the time of beginning of berry
The sum of average temperature °C for 2010- ripening (veraison), the same for the all clones.
2016 ranged from -4.3°C in January to 134.7°C In comparison with Cabernet Sauvignon 30 Vl.
in September. which reached the maturation of the grapes at
Compared to multiannual averages, the 2016- the end of September, the other two ended the
2017 wine years can be characterized as a year maturation 6 - 7 days early, in the first decade
with reduced heliothermal resources on the of October.
background of rich water resources. The low Related to the climatic conditions of 2017 year,
heat regime recorded in April (°tactives = the budding occurred between 07 April and 27
226.7°C) caused a late start (about 10 days) and April, 50% of the buds being in green shoot tip
uneven vegetation in vineyard, a delay that was stage (stage C of Baggiolini, stage 7 to 9 of
not recovered in May and June. BBCH scale) (Baggiolini, 1952) were
The average annual air temperature is 14.5°C registered ten days later in comparison with
and the monthly average is negative only in 2016 year. Little differences were observed in
January (-3.3°C). The 18.4°C blooming the duration of budding and blossoming phases
temperature, the average July temperature of for the three evaluated clones. Difference was
21.2°C, the average August maximum registered from ripening of grapes and harvest,
temperature of 31.6°C and the average air full maturity of the berry being reached at the
humidity in August at 1:00 p.m. of 46-48% end of September for Cabernet Sauvignon 30
263
Vl. and seven days later in case of the other two clones are characterized by medium fertility
clones. and productivity (Table 1).
Fertility and productivity Diseases tolerance

The high percentage of fertile branches in case According to the OIV norms, the Cabernet
of Cabernet Sauvignon 54 Mn. and Cabernet Sauvignon clones presented a good level of
Sauvignon 131 St. ranged values between resistance to Mildew and a very good level to
70.90% and 77.08%, and Cabernet Sauvignon the attack of Oidium and Botrytis cinerea
30 Vl., with values higher than 82%. All three (Table 2).
Table 1. The fertility and productivity of the Cabernet Sauvignon clones in the Valea Calugareasca vineyard
Fertile branches Relative fertility Absolute fertility Relative productivity Absolute productivity
Clones (%) coefficient coefficient coefficient coefficient
2016 2017 2016 2017 2016 2017 2016 2017 2016 2017
Cabernet Sauvignon 30 Vl. 82.40 82.12 1.05 1.03 1.50 1.26 111.49 153.88 159.57 188.09
Cabernet Sauvignon 54 Mn. 70.90 71.80 1.07 0.98 1.50 1.37 140.14 131.52 196.85 188.69
Cabernet Sauvignon 131 St. 75.50 77.08 1.20 1.04 1.59 1.36 108.69 138.49 143.71 180.36
The degree of sugar accumulation was close to
Table 2. Resistance to diseases at the Cabernet Cabernet Sauvignon clones (between 2.38 and
Sauvignon clones in Valea Calugareasca vineyard 2.63 g L-1/day). Acid metabolism was averaged
(years 2016 - 2017)
Clones Diseases Attack level (%) 1.59-1.73 g L-1/day.
Leaves Grapes Compared to the other two clones, the average
2016 2017 2016 2017 weight of a cluster at Cabernet Sauvignon 30
Cabernet
Sauvignon
Mildew
Oidium
1,05±0.1 1.8±0.1
0,88±0.1 0.8±0.1
0,94±0.1 1.4±0.1
0,72±0.1 0.9±0.1
Vl. was higher with 18.92% - 31.06% in the
30 Vl. Botrytis - - 1,00±0.1 0.6±0.1 climatic conditions of 2016 year and with
Cabernet Mildew 0,77±0.1 1.9±0.1 1,15±0.1 1.3±0.1 11.04% - 13.11% in 2017. Cabernet Sauvignon
Sauvignon Oidium 0,95±0.1 0.8±0.1 0,99±0.1 0.8±0.1
54 Mn. Botrytis - - 0,98±0.1 0.5±0.1 30 Vl. was distinguished by a greater average
Cabernet Mildew 0,92±0.1 1.8±0.1 1,30±0.1 1.5±0.2 weight of the berry (126.92 g - 145.44 g)
Sauvignon Oidium 0,9±0.16 0.9±0.1 0,76±0.1 0.7±0.1
131 St. Botrytis - - 0,95±0.1 0.5±0.1 compared to Cabernet Sauvignon 54 Mn. and
Cabernet Sauvignon 131 St. (88.11 g - 128.68
Grapes quality g) (Table 3).
As a result of the evolution of climatic factors The differences in berry weight had an effect
in the maturation period, weight grains were on their structure (Table 4). At Cabernet
11.17% for the Cabernet Sauvignon 30 Vl. Sauvignon 30 Vl. the skin ranged between,
clone and a maximum of 45.95% at the clone 16.4 - 18.6% and seeds between 4.2 - 4.7%,
Cabernet Sauvignon 131 St. correlated with a higher content in pulp (77.2-
78.9%).
Table 3. Mechanical analysis of grapes at harvest

Grape Cluster Berry
Clone Weight (g) Volume (ml) Weight (g) Weight (g)
2016 2017 2016 2017 2016 2017 2016 2017
Cabernet Sauvignon 30.Vl 130.97 149.40 120 136.67 4.05 3.97 126.92 145.44
Cabernet Sauvignon 54 Mn. 106.18 133.79 98.33 118.33 3.84 4.27 102.35 129.52
Cabernet Sauvignon 131 St. 90.29 132.91 78.33 118.33 2.18 4.23 88.11 128.68
Table 4. Berry structure

Skin (%) Seeds (%) Pulp (%)
Clone
2016 2017 2016 2017 2016 2017
Cabernet Sauvignon 30 Vl. 16.4 18.6 4.7 4.2 78.9 77.2
Cabernet Sauvignon 54 Mn. 18.4 22.2 5.1 4.2 76.4 73.5
Cabernet Sauvignon 131 St. 25.8 20.0 5.8 4.8 68.5 75.2
264
Chemical composition of the stum Analysis of the polyphenolic potential of
The potential for the accumulation of sugars in grapes
the stum, a characteristic of the variety, influen- From the point of view of the polyphenolic
ced by the climatic factors of the grape matu- composition, the Cabernet Sauvignon 54 Mn.
ration period, was very high at all genotypes, in clone is evidenced by the total polyphenol
terms of acidity and low pH (Table 5).Grapes index and the anthocyanin potential.
from the vintage 2017 had better sugar The lowest values were obtained for clone 131
accumulation potential due to favourable St. (average data over the two years of study)
weather conditions during the ripening stage. (Figure 1).
Table 5 Chemical composition of the stum

Weight of 100 Volume 100 Sugar (g/l) Acidity pH
Clones berries (g) berries (g/l H2SO4)
2016 2017 2016 2017 2016 2017 2016 2017 2016 2017
Cabernet Sauvignon 30 Vl. 110.86 120.84 100 105 245.9 223.6 3.28 3.61 3.22 2.17
Cabernet Sauvignon 54 Mn. 106.97 108.58 95 95 224.9 219.4 2.95 3.21 2.64 2.29
Cabernet Sauvignon 131 St. 106.68 129.81 95 115 227.9 227.9 2.99 3.36 2.62 2.20
Total polyphenolic index (IPT) 764

800
63 646
700
63 600
449
62 500
61 400
59 255
300 215
60 150
59 200
59
100
58
0
57 Cabernet Cabernet Cabernet
56 Sauvignon 30 Sauvignon 54 Sauvignon 131
Cabernet Cabernet Cabernet Vl. Mn. St.
Sauvignon 30 Sauvignon 54 Sauvignon 131
Vl. Mn. St. Anthocyanins (mg/l) Total anthocyanic potential (mg/Kg)
Figure 1. The polyphenolic potential of grapes (Average values 2016-2017)
The glucoacidimetric ratio records very high The degree of correlation shows a very high
values of exceptional years (over 60%). The value in the case of the Botrytis attack (Figure
increase in this ratio is mainly due to the very 4), the value of the total polyphenolic index
low value of total acidity. being determined almost entirely (99 %) by the
Between the degree of tolerance to cryptogamic frequency of the attack (Urucu, 2014).
diseases and the values of total polyphenol
index (IPT) was recorded a very tight 95
correlation. 85
The degree of mildew attack on grapes (Figure 75
2), with values ranging between 1.3% and
IPT
65
1.5%, influenced the value of the total poly- y = -135x + 256,89
phenolic index (IPT) of the grape must at 55 R² = 0,7961
harvesting by almost 80%, the value of the 45
coefficient of determination R squared being 1,2 1,3 1,4 1,5 1,6
0.7961. LA% Grape Mildew
The intensity of the Oidium attack influenced
Figure 2. The Polyphenol Index (IPT)
the value of the total polyphenolic index to in Grape Mildew
almost 90% (R² = 0.898) (Figure 3).
265
95 ACKNOWLEDGEMENTS
85
This study was funded by Ministry of Agriculture
and Rural Development, Romania; within the
75 Sectorial research and development plan - ADER -
Project 325 „Diversification of the viticultural
IPT
65
assortment for table and wine grapes”.
y = -35,476x + 186,14
55
R² = 0,898
45
REFERENCES
2,5 3 3,5 4
IA% Grape Oidium Baggiolini M., 1952. Les stades reperes dans le
developpement anual de la vigne. Revue Roman de
Figure 3. The Polyphenol Index (IPT) in Oidium attack d'Agriculture, de Viticulture et d'Arboriculture,
Lausanne, v. 8, 4-6.
Dejeu L., 1986. Cercetări privind influenţa solului asupra
calităţii strugurilor la soiul Cabernet Sauvignon.
95 Lucrări ştiinţifice IANB, seria B, vol. XXIX.
Fidelibus M., Christenson L., Katayama D., Verdenal P.,
85
2006. Yield components and fruit composiotion of
75 six Cabernet Sauvignon grapevine select in the
Cenral San Joaquin Vally Californija. Journal of the
IPT
65 American Pomological society, 60 (1): 32-36.

y = -10,23x + 211,11
55 R² = 0,9921 Jones G.V., Davis R.E., 2000. Climate Influences on
Grapevine Phenology, Grape Composition, and
45 Wine Production and Quality for Bordeaux, France.
11 12 13 14 15 16 American Journal Enology and Viticulture, 51 (3):
FA % Grape Botrytis 249-261.
Matei P.M., Dejeu L., Mereanu D., 2009. Research
Figure 4. The Polyphenol Index (IPT) in Botrytis attack concerning the influence of climate change on
grapevine. Bulletin of University of Agricultural
Sciences and Veterinary Medicine, Cluj-Napoca,
CONCLUSIONS Horticulture. 66 (1) 352-358, ISSN/ISBN 1843-
5254. [JOUR: VitisVea W0 10 1788].
The development of the vegetative pheno- Pop N., 2000. Curs de viticultură generală, Ed.
AcademicPress, Cluj-Napoca. ISBN 978-973-757-
phases depends on the climatic conditions 374-2.
during 2016 and 2017 harvest. Differences Stefanini M., Iacono F., Colugnati G., Bregnant F.,
were registered concerning the maturation of Crespon G., 2000. Adaption of same cabernet
the grapes which occurred 6-7 days later in sauvignon clones to the environmental conditions of
case of Cabernet Sauvignon 54 Mn. and north-eastern Italian growing areas. ISHS Acta
Horticulture, 528: 779-784.
Cabernet Sauvignon 131 St. Șerdinescu A., Pîrcălabu L., Ion M., 2006. Influence of
Cabernet Sauvignon 30 Vl. highlighted by a some technological links on the evolution of the
greater weight of the cluster and berries. main physiological and biochemical processes
The potential of sugar accumulation in the must implied in building up grape yield quality at
was high in case of all clones, while the acidity Cabernet Sauvignon cultivar, XXIX World Congress
of Vine and Wine.
and pH values were low. Tebeica V., Popa C., 2005. Results concerning applying
Concerning the polyphenolic composition, of clonal selection of variety Sauvignon, The
Cabernet Sauvignon 54 Mn. was noted by total Romanian Society of Horticulturists, “Ion Ionescu
polyphenolic index (IPT) and Anthocyanin from Brad” Publishing House, Iasi, Year XLVIII
potential. The degree of correlation between Vol. I (48) ISSN 1454-7376.
The Catalogue selected wine grape varieties and clones
polyphenolic index and frequency of the mains cultivated in France, 2009. Eds. Boidron, R. etol.
diseases attack shows a very high value in the Ministry of Ag. Fisheries and Food., 192-193.
case of the Botrytis attack (99%). Urucu J., 2014. Teza doctorat: Studii comparative ale
Further studies will be done concerning the unor clone de Caberent Sauvignon altoite pe diverși
quality of wines in order to establish the degree portaltoi in conditiile Centrului viticol Bolovanu-
Sâmburești. USAMV București.
of maintenance of wines tipicity.
266
TEMPERATURE AND RAINFALL INFLUENCE ON SHOOT LENGTH IN

PINOT NOIR, MERLOT AND CABERNET SAUVIGNON VARIETIES
Eleonora NISTOR1, Alina DOBREI1, Alin DOBREI1, Dorin CAMEN1,

Giovan Battista MATTI2
1
Banat University of Agricultural Sciences and Veterinary Medicine “King Michael I of Romania”,
Faculty of Horticulture and Forestry, Calea Aradului 119, Timisoara, Romania
2
Universita degli Studi Firenze, Dipartimento Di Scienze Produzioni Agroalimentari E
Dell'ambiente, Viale Delle Idee, 30, 50019 Sesto Fiorentino, Italy
Correspondent author email: [email protected]
Abstract
Climate changes from last decades influenced plants growth including grapevine. The objective of the research was to
evaluate the influence of temperature and rainfall on shoot growth in Pinot Noir, Cabernet Sauvignon and Merlot
varieties located in two vineyards, Recaș and Buziaș-Silagiu, during five years (2011-2015). The vines were trained on
vertical trellis by bi-lateral cordon and vertical shoot positioned. For research were selected the main shoots and
lateral shoots from 25 vines from each variety. Shoots length was measured from the base to the growing tip. Shoots
cease growth earlier in dry years 2012 and 2015 compared with the wettest years 2013 and 2014. The warm weather
and moderate rainfall in 2011 favor shoots growth in all three varieties and both locations. Measurements show
significant differences between shoots length. The longest shoots were found in Cabernet Sauvignon variety in Buziaș-
Silagiu vineyard and the shortest in Pinot Noir variety from Recaș vineyard. Results also show that vines subjected to
water stress and high temperatures had shorter main and lateral shoots.
Key words: grapevine, length, rainfall, shoots, temperature.
INTRODUCTION the shoot growing continues the canopy will

need more green pruning to improve berries
Leaf area from canopy have a major development and ripening, including leaf
importance in photosynthesis and therefore in removal (Kliewer and Dokoozlian, 2005). The
vine development, creating the microclimate vine shoot length is important because they
for grape berry development and ripening have poorly developed mechanical tissues, and
(Andreini et al., 2009; Deloire A., 2009). An when they reach a certain length, more than 40-
excessive shoot length and dense canopy 60 cm, can no longer sustain themselves, bends
influence negative the grape veraison and over, which favor the pests and diseases, and
ripening by shading and increase the humidity prevents further maintenance work in vineyards
(Fournioux, 1997; Keller, 2015). Dense canopy (Sánchez and Dokoozlian, 2005).
favours diseases as rots and mildew. Canopy Most vineyards around the world are found in
includes the shoots, leaves and flowers the strip drawn by parallels 30 and 50 in both
respectively berry bunches (Petrie et al. 2000; northern and southern hemispheres (Keller et
Dobrei et al., 2016b). Vine canopy depends on al., 2005). Within these areas, climate has a
grapevine variety, cane pruning, plant vigor, great influence on the development of
growing area and is described by number of vineyards (Santesteban et al., 2010). The vine
shoots/trunk, length, width, height and leaf area is quite resistant to drought; works well in areas
(Dokoozlian and Kliewer, 1995). Shoots with annual rainfall of 500-650 mm, but like
emerge in the spring from buds and grow fast most plants, vines need a decent amount of
before bloom due to the energy reserves from water to survive and grow (Smart et al., 1991;
roots and then their growing slow down as Schultz and Matthews, 1988).
grape berries develops (Lorenz et al., 2005; Each physiological phenomenon is conditioned
Dobrei et al., 2014). Usually in well-balanced by reaching a certain level of temperature and
vineyards, shoots growth stop near veraison; if humidity that marks the beginning or end of a
267
biological stage (Hendrickson et al., 2004). the growing tip, in the first three stages: before
Favorable temperature is correlated with a large bloom, full bloom and fruit set, until the
amount of fruitful buds and more leaves (Gris summer trimming. Measurements were made
et al., 2010). Both spring rainfall and higher using a metric tape. In both vineyards the vine
temperatures greatly contribute to keeping the management was uniform. In recent years, the
plant vigorous (Matthews et al., 1987). temperature increased from the beginning of
The objective of the work was to evaluate the the spring, while rainfall are very heavy or are
temperature and rainfall influence on main and very small quantity in summer; often very wet
lateral shoots growth in first three phenological and hot years (like 2014) alternate with very
stages in three grapewine varieties from warm and dry years. From one area to another,
Buzias-Silagiu and Recas vineyards, in the west during the winter, cold days alternate with
of Romania area, during climate changing years frosty periods. Climatic variability influences
2011-2015. blooming, growth and grapevine development,
grape production and its quality, through the
MATERIALS AND METHODS late spring frosts, high summer temperatures,
heavy rains or hailstorms.
The experiment was carried out in two well- In May 2011 the average temperature was
known vineyards from west of Romania: Recas 0.1°C above normal. From May 1-8, the
and Buzias-Silagiu, during 2011-2015. All weather was colder than usual. Monthly rainfall
three varieties (Pinot Noir, Merlot and Cabernet was 18% lower than normal. There were heavy
Sauvignon) were trained on vertical trellis by rains (on May 24, 25 and 28) often associated
bi-lateral cordon and vertical shoot positioned. with thunderstorms and hail. In June the
In each variety were selected for measurements average temperature was 1.1°C higher than
the main and two lateral shoots from 25 vines. normal. During June 19-24, the weather was
Grapevine shoots monitoring begun each year very hot. Between 25 and 30 June the
in the last decade of April and ended in July, temperature dropped significantly, reaching
during all five years of experiment. Plant below normal values for this period. In July the
material was selected from vines of 6 year-old, average temperature was 1.2°C higher than
planted on east-west orientated rows. Lateral normal. From 9 to 20 July, the weather was
shoots were selected from node positions 2-3. very hot (Figure 1).
Shoots length was measured from the base to
40 90
80
35
70
30
60
25
50
Temperatures (oC)
Rainfall (mm)
20 40
15 30
20
10
10
5
0
0
-10
I II III IV V VI VII VIII IX X XI XII
-5 -20
max (t°C) 2011 average (t°C) 2011 min (t°C) 2011 rainfall (mm) 2011
Figure 1. Temperatures and rainfall (2011)
268
Among 13-18 May 2012, the weather becomes higher than normal standard. In 13-15 July and
very cold in the west. There were heavy rains, 28-30 July, the weather was very hot. Monthly
often thunderstorms and hail. In June, the rainfall was 48% below standard normal. May
average temperature was 3°C above normal. 2012 was characterized by a higher air
Monthly rainfall was 38% below standard temperature than normal. Often thunderstorms
normal. In July, the temperature was 4.5°C and hailstorms were recorded (Figure 2).
50 140
40 120
30 100
Temperatures (oC)
Rainfall (mm)
20 80
10 60
0 40
-10 20
-20 0
The rainfall between 1st and 31 May (2013) temperature values ranged from 17 to 29°C
was normal (26-50 l/m2) in the north of Banat during the warmest periods. During June, there
and around 51-100 l/m2 in the other areas of the were rainy days, thunderstorms and hailstorms
region. In June 2013 normal thermal days that partially/totally affected the vine by
alternate with periods with higher air breaking canes and shoots (Figure 3).
temperature than usual. The average daily air
45 140
40
35 120
30
25 100
20
Temperatures (oC)
15
Rainfall (mm)
80
10
5
60
0
-5
-10 40
-15
-20 20
-25
-30 0
269
In May 2014 normal days alternate with Romania. June 2014 had warmer days, with
intervals in which the air temperature was periods in which the air temperature was
lower than usual. Precipitations were among normal. During June 2014 heavy rainfall were
51-100 l/m2, in the south of Timis County, recorded in large areas of the western part of
abundant (101-125 l/m2) and even excessive the country, which led to an improvement of
(126-212 l/m2) on large area in west of the soil moisture.
45 140
40
35 120
30
100
25
Temperatures (oC)
20
Rainfall (mm)
80
15
10
60
5
0
40
-5
-10 I II III IV V VI VII VIII IX X XI XII 20
-15
-20 0
40 60
35
30 50
25
20 40
Temperatures (oC)
15
Rainfall (mm)
10 30
0 20
-5
-10 I II III IV V VI VII VIII IX X XI XII 10
-15
-20 0
Figure 5. Temperature and rainfall (2015)
Quantities of water in the interval 01-30 June extended surfaces and very high (01-150 l/m2),
2014 were reduced (12-50 l/m2), in the western in the south. Weather conditions were favorable
Timis County, normal (51-100 l/m2) on for vine growth and development in the vine.
270
First decade of July 2014 was warm weather, and Merlot varieties, from Buzias-Silagiu and
after which the temperature increased Recas vineyards, among 2011 and 2015, are
gradually. The higher temperature ranged shown in Figure 6. In Pinot Noir the longest
between 19 ... 35°C and the lowest between 10 main shoot before bloom was in 2014 (27 cm)
... 26°C. There were heavy rainfalls but also when were recorded daily temperatures of 310C
thunderstorms, and isolated hailstorms. The and nights with no less than 70C. The same
rainfall in the period 01-31 July 2014 was behavior was observed in the other two
among 51-100 l/m2, in the southwest and varieties, Cabernet Sauvignon (64 cm in 2014)
central of the Timis County. On extended areas and Merlot respectively (49 cm). The spring of
rainfall were heavy (101-125 l/m2) and over 2015 (similar to 2011) was cool, with
normal (126-241 l/m2), which favored the temperatures slightly below the average of the
downy mildew (Plasmopara viticola) and period, which delayed the blooming with about
powdery mildew (Uncinula necator) (Figure 7-10 days without major influence on the vine
4). In May 2015, in the center of the region, phenology. However the shortest main shoots
normal (26-50 l/m2), high (51-100 l/m2), before bloom, in all three varieties, were
abundant (101-125 l/m2) and excess rainfall registered in 2015 followed by results from
(126-200 l/m2) were reported on extended areas 2011.
of Banat. Moisture in the 0-100 cm of soil at Higher grow rates were observed before full
the end of May 2015, in Timis County, was bloom, followed by slow grow after fruit set
low values. The summer season of 2015 was due to the competition for nutrients and water
characterized by a warmer weather than among canopy and bunches (Figure 6). In full
normal. Between May and June, the amount of bloom stage, the longest main shoot was
rainfall was deficient (< 150 l/m2) (Figure 5). recorded in Cabernet Sauvignon (104 cm) in
Data were subjected to statistical analysis using 2014. In fruit set stage, remains the same rank
Statistica 13.0.159.7 software for Windows of main shoot length for all varieties. Young
(One way ANOVA). shoots were green, healthy and vigorous, and
flower bunches equally distributed on the
RESULTS AND DISCUSSION canes. In 2014 the constant rainfalls at regular
intervals, alternating with sunny and low wind
Results concerning the length measurements of days helped to air the vineyard and decreased
main shoots in Pinot Noir, Cabernet Sauvignon humidity around flowers.
120 120
110 110
100 100
90 90
80 80
70 70
(cm)
(cm)
60 60
50 50
40 40
30 30
20 20
10 10
0 0
2011 2012 2013 2014 2015 2011 2012 2013 2014 2015 2011 2012 2013 2014 2015
Pinot Noir Cabernet Sauvignon Merlot
Main shoot /Before bloom/Buzias Main shoot /Before bloom/ Recas

Main shoot /Full bloom/ Buzias Main shoot /Fruit set/ Buzias
Main shoot /Full bloom/Recas Main shoot /Fruit set/ Recas
Figure 6. Main shoot length before bloom, full bloom and fruit set stages, in Pinot Noir, Cabernet
Sauvignon and Merlot varieties, from Buzias -Silagiu and Recas vineyards (2011-2015)
271
Cheng et al. (2014), in a study concerning the was quite similar for around 130 days, until
influence of soil properties and climatic ripening stage. The final main shoot length was
conditions of 2011 and 2012 years, on Cabernet in average 3.22 m for Merlot variety and 2.90
Sauvignon variety, found that the average shoot m for Cabernet Sauvignon. Growing rate
length at harvest, in two vineyards from China, observed was 5.0 cm per week before blooming
was between 122 and 136 cm, length that can and around 25.0 cm per week after until grow
be reached and exceeded by main shoots of decrease after fruit setting. In Leal G.R. (2007)
Cabernet Sauvignon from Buzias-Silagiu and studies, on Pinot Noir variety from New
Recas until the harvest time. Zealand, main shoot final growth was 89.1 m.
Sabbatini and Schilder (2012) found in their Hunter and Visse (1990), in their research
research from Michigan vineyards, longest developed in South Africa, concerning the
main shoot in Pinot Noir variety in all three effect of defoliation on Cabernet Sauvignon
stages: 67 cm (before bloom), 76 cm (full growth, found longer lateral shoot length in
bloom) and 95 cm (fruit set) respectively. berry set stage, amongst 63 - 91 cm, very
Reynolds and Naylor (1994) investigate Pinot significant longer compared to lateral shoots of
Noir variety in a glasshouse situated in British Pinot Noir from Buzias-Silagiu or Recas
Columbia, Canada. vineyards. Longer lateral shoots are the result
Lateral shoots measured between 47 and 108 of partial defoliation which improved the light
days after full bloom was among 6 and 143 cm. environment and leaves photosynthesis in the
Lateral shoots from the Pinot Noir grown in vine canopy (Dobrei et al., 2016c).
both vineyards from the west of Romania are Merlot is a medium - large vigor. It is adapted
included in these limits and are much shorter. to different soil and climate except arid areas.
Schreiner et al. (2013), studied the impact of Variety resistance to frosts is low (-16°C -
NPK supply on leaf area and shoot growth 18°C) and drought, and medium to diseases.
before bloom stage in pinot Noir variety. Sprouting takes place early, and is
According to their research results, the shoot consequently sensitive to spring frosts
length was between 84-117 cm, and little Unfavorable weather during flowering leads to
influenced by different NPK supply. millerandage (Dobrei et al., 2015).
Pinot Noir is a medium vigor variety and Variety is also susceptible to mold (although it
temperatures above 350C in 2014 and 2013 has a better mold resistance than other
summer days slow the shoot growing due to the varieties) (Robinson J., 2003). It is a poorly
shutting down of the photosynthesis. tolerant to drought and frost-resistant (-180C)
Cabernet Sauvignon has large vigor and (Dobrei A., 2004). Similar results to those from
medium fertility (65-70% fertile shoots). It has Buzias-Silagiu and Recas vineyards,
good tolerance to frost (-20, -220C), very concerning the main shoot length in Merlot
resistant to drought, good tolerance to oidium, variety (from 37 to 90 cm) were found in two
gray mold and rots (Dobrei et al., 2016b). trials conducted during 1995-1998 in the
The vegetative vigour of Cabernet Sauvignon Research station Agroscope Changins-
from high altitude vineyards of Southern Brazil Wädenswil from Switzerland.
was studied by Rufato et al. (2014) after Jemini et al. (2010) found a shoot length in
application of prohexadione-calcium for Merlot variety affected by downy mildew,
reducing shoot growing. Despite the treatment, between 12.30 cm in the second decade of May
the main shoot final length range among 209.7 and 81.99 cm after 30 days in June in the 1996
and 258.1 cm). trial and between 25.91 and 78.59 in 1998.
During 2006 -2009, Borghezan et al. (2012), Lateral shoots length before bloom, full bloom
evaluate shoot growth in Merlot and Cabernet and fruit set stages, in Pinot Noir, Cabernet
Sauvignon, cultivated in São Joaquim Sauvignon and Merlot varieties, from Buzias-
vineyards from Brazil. They found out that the Silagiu and Recas vineyards (2011-2015) are
main shoots average length in both varieties shown in Figure 7.
272
40 50
45
35
40
30
35
25 30
(cm)
(cm)
20 25
20
15
15
10
10
5 5
0 0
2011 2012 2013 2014 2015 2011 2012 2013 2014 2015 2011 2012 2013 2014 2015
Secondary
Pinot Noirshoots /Before bloom/ Buzias Secondary shoots /Before bloom/
Cabernet Sauvignon Recas
Merlot
Secondary shoots /Full bloom/ Buzias Secondary shoots /Full bloom/ Recas
Secondary shoots /Fruit set/ Buzias Secondary shoots /Fruit set/ Recas
Figure 7. Secondary shoots length before bloom, full bloom and fruit set stages, in Pinot Noir,
Cabernet sauvignon and Merlot varieties, from Buzias-Silagiu and Recas vineyards (2011-2015)
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274
VOLATILE PROFILE OF FETEASCĂ NEAGRĂ WINES OBTAINED

IN MURFTALAR REGION AND THE INFLUENCE OF THE ORGANIC
AND CLUSTER THINNING VITICULTURAL PRACTICES
Victoria ARTEM1, Arina Oana ANTOCE2

1
Research Station for Viticulture and Oenology Murfatlar, 2 Calea București,
905100, Murfatlar, Constanta
2
University of Agronomic Sciences and Veterinary Medicine of Bucharest, Faculty of Horticulture,
Abstract
‘Feteasca neagră’ is a versatile grape variety, with results highly dependent on the cultivation region and viticultural
practices. In Murfatlar this variety is cultivated both organically and conventionally, thus providing a good opportunity
to study the behaviour of the variety in the two cultivation systems. Furthermore, in both culture systems, in some
experimental variants 30% of the grapes from each vine were removed after the cluster formation - a practice often
employed with an aim to obtaining a better-quality crop. The grapes harvested in autumn at technological maturity
were vinified in 4 variants, in accordance to the viticultural practices applied: organic and conventional cultivation,
with or without cluster thinning. In this work the influences of these viticultural practices on the volatile profile of the
obtained wines were evaluated by using a flash GC-electronic nose. Although some differences are present when 30%
cluster thinning is applied, the main influence in the volatile profile of wines is still induced by the cultivation system,
irrespective of the grape reduction practice. The variability among the volatile profiles of the 4 groups of wines is
explained in proportion of 85.3% by the cultivation system and only 12.4% by the grape reduction practice. The wines
made from conventionally grown grapes display a more complex volatile profile as compared to the wines obtained
from organic grapes.
Key words: ‘Fetească neagră’, electronic nose, volatile profile, organic viticulture, cluster thinning.
INTRODUCTION Murfatlar vineyards, consisting of the reduction

of the clusters on each vine by 30%. While
‘Fetească neagră’ is a versatile grape variety, some improvement in polyphenolic quality was
with results highly dependent on the cultivation obtained by this cluster thinning (Artem et al.,
region and viticultural practices. Considering 2015), the organic cultivation also showed
the increasing consumption of organic wine some increase in phenolic compounds
and the constant demand for quality wines, it is important for health (Artem et al., 2014b) and
logical that practices such as organic wine ageing.
cultivation or the reduction of yield by cluster However, both practices are costly and should
thinning practice should be taken into account. be applied only when the wines can be
Due to the particularities of the Murfatlar perceived by the consumers as being different.
region, ‘Fetească neagră’ finds here conditions For this reason, comparing the volatile profiles
to be cultivated not only by the conventional with an electronic nose based on flash
technology, but also organically. In previous chromatography is an easy way of determining
studies (Ranca et al., 2010), ‘Fetească neagră’ if the wines resulted from grapes produced with
of Murfatlar showed that it is suitable for different viticulture technologies can be clearly
organic cultivation, several good results being discriminated.
already reported (Artem et al, 2014a). Even though the taste of wine is not evaluated
Also, to improve the concentration of the in this work, the volatile profiles determined
anthocyanins accumulated in the grapes, a can explain some of the effects induced by the
special practice, used by other authors too viticulture technologies and could also be a
(Bubola et al., 2011), was applied in the basis for further analysis.
275
MATERIALS AND METHODS For the electronic nose analysis, 4 ml samples
from each wine bottle were taken in 3 vials of
In Murfatlar, ‘Fetească neagră’ variety is 10 ml volume. Thus, each group of wines
cultivated both organically and conventionally, consisted in 9 analyzed samples.
thus providing a good opportunity to study the
behaviour of the variety in the two cultivation RESULTS AND DISCUSSIONS
systems. The technologies for the cultivation in
both systems are those usually applied in the In order to separate the groups of wines based
region and they were detailed elsewhere on some of the chromatographic peaks
(Artem, 2017). recorded by the electronic nose, from among all
In order to assess if a better quality of grapes the peaks observed only some were selected,
can be obtained by reducing the yield, in both namely those with a discrimination power
culture systems, variants with 30% of the grape above 0.5 (more exactly from 0.835 down to
reduction were also produced. The grape 0.497). Also used were those peaks with lower
thinning was performed on each vine, 30% of overall discrimination power but automatically
the clusters being removed after the cluster selected by the software of the apparatus as
formation. being of importance.
The grapes harvested in autumn at As a result, the four groups of wines were
technological maturity were vinified in 4 separated by the electronic nose based on the
variants, in accordance to the viticultural multivariate statistical analysis of the
practices applied: organic and conventional discriminant factors (Figure 1).
cultivation, with or without cluster thinning.
The grapes obtained from conventional and organic, conventional,
30% thinning 30% thinning
organic growth system, both from vines in
which grape thinning was practiced or not,
were prepared in triplicate.
organic, conventional,
The resulted wines are coded as described in no no thinning
Table 1: thinning
Table 1. ‘Fetească neagră’ wine samples
Wine code
Organic Conventiona Grape Group Figure 1. Discriminant Factor Analysis (DFA) diagram
cultivation l cultivation thinning Colour of the ‘Fetească neagră’ wine groups obtained from
FN_Eco_B1_M light
grapes with different cultivation technologies (light blue
FN_Eco_B2_M x blue
FN_Eco_B3_M = organic cultivation; dark blue = organic cultivation and
FN_Eco_B1_R dark grape thinning, light brown = conventional cultivation;
FN_Eco_B2_R x x blue dark brown = conventional cultivation and grape
FN_Eco_B3_R thinning)
FN_Con_B1_M light
FN_Con_B2_M x brown
FN_Con_B3_M
FN_Con_B1_R dark In Figure 1 it can be seen that the groups of
FN_Con_B2_R x x brown wines were separated mostly on the basis of
FN_Con_B3_R
DF1 axis (responsible for 85.26% of the
variability observed), which represents the
The influences of these viticultural practices on cultivation system (organic towards the left
the volatile profile of the obtained wines were side of the diagram and conventional - towards
evaluated by using a flash GC-electronic nose the right side of the diagram). The grape
(Heracles I, Alpha-MOS, France) equipped thinning accounted only for 12.38% of the total
with two short chromatographic columns of variability, but still, it can be said that this
different polarities (1A: DB5 and 2A: viticultural practice did have a specific
DB1701). The equipment, software and the influence on the resulted wines.
method applied are described in detail in More specifically, the separation of the groups
previous papers (Antoce and Namolosanu, of wines from organic and conventional grapes,
2011; Antoce 2012a, b; Antoce, 2013; Antoce respectively, was determined by certain volatile
et al., 2015; Antoce and Cojocaru, 2017). compounds. In Figure 2, the vectors
276
corresponding to the selected discriminant Table 2. Volatile compounds found to be discriminant
chromatographic peaks are also included in the for the wines obtained with ‘Fetească neagră’ grapes
conventionally cultivated
diagram and, as it can be seen, the loadings of
Column Peak Discri- Compound Type of
most of these vectors on the horizontal axis (retention mination identified aroma*
tend to be higher for the wines from time) Power
conventional grapes. This also suggests an 1A (DB5) 8.49 0.557 acetic acid acid, fruit,
pungent,
increased aromatic complexity of those wine sour, vinegar
samples 1A (DB5) 8.99 0.556 not
identified
1A (DB5) 10.71 0.497 ethyl apple, butter,
butyrate cheese,
organic, conventional, pineapple,
30% thinning strawberry
30% thinning
1A (DB5) 11.90 0.612 2-methyl apple, fruit,
ethyl pineapple
butyrate
1A (DB5) 15.94 0.633 3-hexen-1-ol green
organic,
conventional, 1A (DB5) 16.50 0.529 amino- -
no benzaldehyd
thinning no thinning e
1A (DB5) 18.69 0.835 3-hexenyl banana,
acetate candy, floral,
green
1A (DB5) 19.49 0.667 ethyl apple peel,
hexanoate brandy,
Figure 2. Discriminant Factor Analysis Diagram of the overripe fruit,
‘Fetească neagră’ wine groups with the discriminant pineapple
peaks (light blue = organic cultivation; dark blue = 1A (DB5) 21.67 0.578 not
identified
organic cultivation and grape thinning, light brown = 1A (DB5) 23.91 0.518 2-isopropyl- green pepper;
conventional cultivation; dark brown = conventional 3- bell pepper
cultivation and grape thinning) methoxypyra
zine
2A 12.14 0.663 close to
(DB1701) trans-3-
With the use of the Arochembase software of hexen-1-ol
2A 20.22 0.562 3-hexenyl banana,
the electronic-nose, some of the compounds (DB1701) acetate candy, floral,
corresponding to important discriminant peaks green
2A 22.44 0.758 2-phenyl- fruit, honey,
were identified. (DB1701) ethanol lilac, rose,
Table 2 contains the volatile compounds wine
identified for the peaks found to be *The flavour profiles of the identified substances, included in the table
to generally describe the possible expected aroma, are taken from
discriminating for the wines obtained with Pubchem Open Chemistry database (https://pubchem.ncbi.nlm.nih.gov).
grapes from the conventional culture with or
without grape thinning. The volatile compounds mostly correlated to
As it can be seen, the specific discriminant wines from organic culture are included in
volatile profile of the wines obtained from Table 3.
conventional grapes is quite complex, with
several compounds conferring fruity aroma, Table 3. Volatile compounds found to be discriminant
with some green and floral notes too. for the wines obtained with ‘Fetească neagră’ grapes
Although very close to the ‘Fetească neagră’ organically cultivated
conventional group with 30% grape thinning, Column Peak Discrimi- Compound Type of
the group of wines from conventional grapes (retention nation identified aroma*
time) Power
without yield reduction had fewer discriminant 1A (DB5) 10.37 butyl 0.559 apple, banana,
volatile compounds. With a 0.772 acetate glue, pungent
1A (DB5) 32.74 ethyl 0.086 brandy, grape,
discrimination power the peak 14.32-2A (on decanoate pear
DC1701 column) is the only one strongly 2A 11.32 trans-3- 0.271 green
(DB1701) hexen-1-
correlated to conventional ‘Fetească neagră’ ol
without grape reduction. The peak is close to 4- 2A 13.87 ethyl-2- 0.559 apple, ester,
(DB1701) methyl- green apple,
methoxy-2-methyl-2-butanethiol, which is a butyrate kiwi,
compound with aroma of black currant, one of strawberry
the most specific flavours for this grape variety.
277
Basically, the influence of the grape thinning Thus, it may be safely said that the cluster
on the wines form organic grapes are not very thinning techniques applied in the vineyards for
much discriminated. However, with a 0.521 ‘Fetească neagră’ do not sufficiently affect the
discrimination power the peak 32.74-1A (on volatile profile of the final wines, so that the
column BD5) is strongly correlated to FN consumer will most likely not be able to
organic without grape thinning. The peak is perceive a clear difference in the aroma.
close to ethyl-decanoate, which mostly confer
general, non-specific wine/grape aroma.
Overall, the discriminant peaks can
differentiate the groups of wine by statistical
analysis, but the fingerprint of the wines,
consisting of the peaks with their height or
area, are not very different. Figure 3 shows that
the fingerprints of the wines are quite similar,
and especially the wines from grapes where
cluster thinning was performed are not easily Figure 4. Odor distances of ‘Fetească neagră’ wines
differentiated from those produced without groups produced with grapes from different viticultural
thinning. However, the samples from organic technologies (blue = wines from grapes organically
cultivated, brown = wines grapes conventionally
grapes (blue colour on the diagram) have clear cultivated)
different fingerprints from those from
conventionally cultivated grapes (brown colour Aside of the chemical composition of the
on the diagram), irrespective of the application grapes, the winemaking and the post-
of cluster thinning. To verify if the cluster fermentative evolution are also influencing the
thinning practice caused a significant influence final wine profile. For example, the
on the volatile profile of the final wines, the discrimination of the wine groups was initially
distances between the groups of wines were performed based on all the peaks with a
calculated in odor units. Figure 4 shows that the discrimination power over 0.500. However, by
groups from grapes organically cultivated sensory and chemical analysis some samples,
(blue) are relatively separated from those from but not all, were found to have slightly higher
grapes conventionally cultivated (brown), but volatile acidity. To eliminate this bias from the
inside each type of technology, the odor discrimination analysis, the statistical DFA
distances between samples from grapes with analysis was repeated without the peaks related
and without cluster thinning is not significant to acetic acid (volatile acidity found at retention
(groups of light and dark wine samples of the times 8.49 and 8.99).
same colour overlap). Figure 5 (a and b), obtained based on all peaks
with a discrimination power over 0.500, except
those for volatile acidity, shows that the groups
of wines are separated even better than earlier,
still most of the variability being explained by
the culture system (DF1 = 82.22%), while the
grape thinning accounted for 15.9% (DF2) of
the total variability. This slight increase in DF2
(from 12.38% to 15.9%) means that without the
volatile acidity which developed only in some
of the samples after winemaking, the
differences between samples with or without
cluster thinning should be a bit more evident.
However, the discriminant peaks (Figure 5. b)
are not modified, the compounds and their
Figure 3. ‘Fetească neagră’ wines fingerprints based on
the height of the discriminant chromatographic peaks
importance in discrimination remaining the
(blue = samples from grapes organically cultivated, same as described in the tables above.
brown = samples grapes conventionally cultivated)
278
Conferences 9, 02003 (2017), 40th World Congress of
a) Vine and Wine, DOI: 10.1051/bioconf/20170902003,
1-6.
Antoce O.A., Stroe M.V, Cojocaru G.A., 2015. Tentative
application of an electronic nose to the study of the
parentage of Romanian grape varieties Sarba and Alb
Aromat, Agriculture and Agricultural Science
Procedia, Elsevier 6, 110-117.
Antoce O.A., 2013. Application of an electronic nose
based on gas-chromatography to the differentiation of
groups of wines, 36th World Congress of vine and
b)
wine, Vine and Wine between Tradition and
Modernity, Bucharest - Romania, 2-7 June 2013,
Proceedings of the 36th World Congress of vine and
wine, ISBN: 979-10-91799-16-4, Section 2 Enology,
12 pag, 2013-082_Arina-E-nose OIV 2013-final.pdf.
Antoce O.A., 2012a. Results obtained by applying
modern winemaking technologies on the Cramposie
variety cultivated in a new terroir, AgroLife Sci. J.,
Vol. I, 89-92.
Figure 5. Discriminant Factor Analysis Diagram of the
Antoce O.A., 2012b. Study of the possibility of
‘Fetească neagră’ wine groups after the elimination of
discrimination by the electronic nose of Fetească
bias induced by volatile acidity
regală wines produced with neutral and aromatic
yeasts, Scientific papers, Series B. Horticulture,
CONCLUSIONS Volume LVI, Section Miscellaneous, 419-426.
Antoce O.A., Namolosanu C.I., 2011. Rapid and precise
discrimination of wines by means of an electronic
This work shows that the flash GC-electronic nose based on gas-chromatography, Revista de
nose can distinguish wines produced from Chimie 62 (6), 593-595.
grapes obtained by different viticultural Artem V., 2017. The influence of cultivation techniques
practices, simply on the basis of their volatile of red wine grapes and of their processing on wine
quality and on their phenolic potential, Ph.D. thesis,
profile. Although some differences are present
USAMV Bucharest, Chapter 4.1.3.
when 30% cluster thinning is applied, the main Artem V., Antoce O.A., Namolosanu I., Ranca A.,
influence in the volatile profile of wines is still Petrescu A., 2015. The effect of green harvest on the
induced by the cultivation system, irrespective quality of organic grapes cultivated in Murfatlar
of the grape reduction practice. The variability viticultural centre, Bulletin of University of
Agricultural Sciences and Veterinary Medicine Cluj-
among the volatile profiles of the 4 groups of
Napoca. Horticulture,Vol. 72, No. 2, 299-304.
wines is explained in proportion of 85.3% by Artem V., Antoce O.A., Nămoloşanu I., Ranca A.,
the cultivation system and only 12.4% by the Petrescu A., 2014a. Evaluation of quality of
grape reduction practice (these figures change, organically grown grapes in the viticultural center of
but only slightly, when the peaks corresponding Murfatlar. Scientific Papers. Series B, Horticulture,
Bucharest, Vol. LVIII, 113-118.
to volatile acidity are excluded from analysis).
Artem V., Geană I., Antoce O.A., 2014b. Study of
For the particular case of ‘Fetească neagră’ phenolic compounds in red grapes and wines from
produced in Murfatlar region, the wines made Murfatlar wine center, Ovidius University Annals of
from conventionally grown grapes display a Chemistry, Vol. 25, Nr. 1, 47-52.
more complex volatile profile as compared to Bubola M., Persuric D., Ganic K.K., 2011. Impact of
the wines obtained from organic grapes. cluster thinning on productive characteristics and
wine phenolic composition of cv. Merlot, Journal of
Food, Agriculture & Environment, 9 (1), 36-39.
REFERENCES Ranca A., Negraru A., Artem V., Bolos P., 2010.
‘Fetească neagră’ 9Mf, a new grapevine clone of
Antoce O.A., Cojocaru G.A., 2017. Detection with flash quality and productivity, Bulletin of University of
gas chromatography electronic nose of the general Agricultural Sciences and Veterinary Medicine Cluj-
influences of glutathione, ascorbic acid, tannin and Napoca. Horticulture Vol. 67, No. 1, 497.
carbon dioxide treatments on the volatile profiles of
white wines of Fetească regală, BIO Web of
279
280
RESEARCHES REGARDING QUALITY AND QUANTITATIV

PERFORMANCE OF SOME TABLE GRAPES IN THE EXPERIMENTAL
FIELD OF USAMV BUCHAREST
Marinela Vicuţa STROE, Daniel Nicolae COJANU

59 Mărăşti Blvd, District 1, 011464, Bucharest, Romania,
Phone: +40 21 318 36 36/216
Abstract
The main processes of fecundation and pollination on vine are largely influenced by the genetic profile of the variety,
external factors involved like - temperature, precipitation, insolation, relative humidity and also by special crop
techonology applied. The aim of this research was to study some of the most grown table grapes varieties in Romania
(and also worldwide), varieties which belong to a the same kin group - ‘Muscat Hamburg’ cv., ‘Muscat d’Adda’ cv.,
‘Afuz Ali’ cv. and ‘Victoria’ cv. For example, ‘Muscat d’Adda’ cv. has been obtained by self-pollination of ‘Muscat
Hamburg’ variety and ‘Victoria’ variety has been obtained by crossing of two cultivars Cardinal and ‘Afuz Ali’. The
results showed that the quantity and qualitative performance of these varieties, expressed by productive indices,
carpometric values of grapes and berry and also organoleptic qualities has been influenced both by special crop
technology (isolation of inflorescences and pinching fertile shoots a few days before blooming) and by environmental
factors recorded during the study. In the study, has been obtained higher average values of the most analyzed
indicators and the best results have been recorded by: ‘Muscat Hamburg’ cv., ‘Muscat d’Adda’ cv., ‘Afuz Ali’ and
lesser, but also in positive limits, ‘Victoria’ variety.
Key words: cluster, inflorescences, phenology, table grape, yield.
INTRODUCTION and also the ripening of the grapes depends

very much on the factors mentioned above.
Due to unfavourable conditions during
It is well known that the quantitative and
flowering and the way the pollination process
qualitative parameters of the harvested crops
has been carried out, the percentage of fruit
are influenced by the environmental factors,
growth will be different and the fruit set will
like the soil type, the technology applied, the
have a different evolution developing the
genetic nature of the varieties, but from all
millerandage phenomenon (Dobrei et al.,
these factors, the greatest impact has the
2005). This phenomenon is quite common on
climate, (Jones, 2005; Cleland et. al., 2007;
the grape varieties that require foreign pollen,
Gladstones, 2011; Shinomiya et al., 2015; Van
like ‘Muscat Hamburg’, due to the low pollen
Leeuwen and Darriet, 2016). Practically, in the
germination and in this case the growth of
most wine-growing areas, there are recorded
berries it stops at 6-7 mm, due to the lack of
more and more changes in the climate as a
hormone substance, this it may also occur in
results of the high average temperatures, due to
grapes varieties that develop incompatibility of
low precipitations followed by extreme events
its own pollen (Constantinescu et al., 1959;
(Tomasi et al., 2017). Even though, the annual
Lepadatu, 1979; Stroe et al, 2014). After this
sequence of phenological stages of grapevine is
phenomen occurs, the berries start ripening
commonly observed to be accelerated with an
earlier and gain higher quantities of sugar, but,
increase in temperature (Duchêne and
at the time of harvesting, the quality of grapes
Schneider, 2005; Duchêne et al., 2010; Parker
et al., 2013; Blanco-Ward et al., 2017), the is shortcoming, especially in table grapes
varieties. Even more, various studies have
annual cycle of the vine, the flowering, fruit set
shown that gaining excessive sugar, it is not
281
followed anymore by an optimal polyphenolic have been analyzed during 20 may - 20 june
and aromatic ripening; the intake of grapes is 2017, using the daily average from 6 hourly
alterated; in fact, the difference between data (05:00, 08:00, 11:00, 14:00, 17:00, 20:00,
aromatic and phenolic content and tehnological 23:00) registered at Bucharest - Baneasa,
ripening (ratio between sugars/acids) is always Romania. The forecast was performed over a
higher. This may lead to the conclusion that longer period just to ensure a better accuracy of
obtaining well-balanced yield in terms of the results, although the flowering phenophase
quantitative and qualitative parameters is a of the studied varieties lasted 12 days (29 May-
constant challenge for the viticulturist. Based 10 June 2017). For the acknowledgement of the
on this consideration, the aim of this study has flowering phenophase, the study used the
been to analyze the evolution and influence of updated version of the universal scale for the
climatic factors in 2017, in a critical period for description of Monocots and Dicotyledons
the vine (20 May - 20 June), in conjunction numbered 00-97, with a special look at the
with 2 special cultural techniques - the isolation main growth stage 6: Flowering, stages 61 and
of the inflorescences and pinch of the growth 67 and main growth stage 7: Development of
peak of fertile shoots on table grapes. The fruits, 71 and 77 (Pierot and Rochard, 2013).
result of the experience has been analyzed also Three observations have been made at different
by quality parametres of the grapevine products growing stage (Table 1), taking into account all
(yield) influenced by this factors. the inflorescences on 6 vines but the isolation
of the inflorescences in waxed paper bags and
MATERIALS AND METHODS pinching fertile shoots were applied on three
vine a few days before flowering for each
2.1. Plant material and growth conditions variety, resulting in a total of 8 experimental
Four grape varieties were taken in the study: variants. Harvesting of the grapes has been
‘Muscat Hamburg’ cv., ‘Muscat d’Adda’ cv., carried out at the full ripening stage of each
‘Afuz Ali’ cv., from the world collection and variety.
‘Victoria’ cv., variety obtained in Romania in
1978 by ‘Victoria’ Lepădatu and Gh. Condei RESULTS AND DISCUSSIONS
(Table 1). The main updated data on these
varieties can be found in the Vitis International The studied varieties have reacted differently,
Variety Catalog (www.vivc.de). In Romania, the indifferent by cultural applied techniques, in
first three listed varieties occupies the largest terms of the percentage of fruit set, the
areas, although all three varieties have proven percentage of shaking of the formed flowers
over the years to be sensitive to temperature and the manifestation of millerandage
and humidity factors during flowering phenomenon. The distinct character of the wine
phenophase. Grape table varieties are located in year 2017 in terms of climatic peculiarities led
the experimental field of the ampelographic to an early age in terms of flowering onset
collection from the University of Agronomic (Cleland et al., 2007; Stroe et al., 2017), noting
Sciences and Veterinary Medicine of that all analyzed varieties bloomed at the end of
Bucharest. During the study, has been made the third decade of May. The phenophase
basic observations and determinations, period has been lasting for approximately 12
commonly used in the current technology, but days (29 May-10 June 2017), with a gap of 2-3
attention was directed to the carpometric days between varieties, as follows: on ‘Muscat
elements that define the productive and d’Adda’ cv. and ‘Afuz Ali’ cv., blooming
qualitative potential: average weight of a grape started on May 29, and at ‘Victoria’ cv. started
(g), average weight of a berries (g), yield later in two days. During the period of interest,
(kg/vine), 0Brix, (refractometric), total acidity the daily average temperature values have had
(g tartaric acid/L by titration), sugar-acidity significant fluctuations (Figure 1), but the
indices. 18.59ºC average, was even lower than the
2.2. Phenological and temperature data optimum of varieties in the first decade of May,
During the study, meteorological parameters only 15.5ºC, but in full flowering the value was
(temperature, precipitation, relative humidity)
282
19.84ºC, even recording a ceiling of 20ºC and their impact to the studied varieties. The
between 3rd and 8th of June. average value had been 67.25% and the highest
was 91% registered in the first decade of May.
Practically, optimal values, close to the normal
blooming range, 55-65%, had been registered
on 26th of May.
Figure 1. Evolution of the average temperature (°C)

for flowering stage
In this matter, it can be seen that in the period

mentioned above, the first three varieties were
in the process of flowering, being known in the
viticultural practice that a percentage of Figure 2. Evolution of the average relative humidity (%)
20-30% of the total flowers bloom in the first for the period 20 may-20 june 2017
2-3 days of flowering, in the next 3-4 days
In the same period mention above, rainfall
bloom 60-70% and only a small percentage of
records a value of 37 mm, and in full flowering
flowers open at the end of the phenophase.
process, 24 mm (Figure 3).
One important note is that the temperature at
Regarding to the special vine techniques - the
the end of phenophase for ‘Victoria’ variety
isolation of the inflorescences and the pinch of
was only 16°C (June 9), and this resulted in
the growth peak of fertile shoots - applied on
forming of a larger number of uneven berries,
whatever experimental variant had analyzed.
Figure 2 analysis the relative humidity values
Table 1. The genetic origin of studied varieties
Muscat Muscat Afuz

Prime name ‘Victoria’
Hamburg d’Adda Ali
Variety number VIVC 8226 8050 122 13031
Country of origin of the variety U. K. Italy Liban România
Species Vitis vinifera L. Vitis vinifera L. Vitis vinifera L. Vitis vinifera L.
Pedigree as given by Trollinger x Muscat Muscat Hamburg - -
breeder/bibliography Alexandria SP
Pedigree confirmed by markers Schiava grossa x - - Cardinal x ‘Afuz
Muscat Alexandria Ali’
Prime name of pedigree parent 1 Schiava grossa - - Cardinal
Prime name of pedigree parent 2 Muscat Alexandria - - ‘Afuz Ali’
Year of crossing 1850 - - 1964
Last update 18.01.2018 18.01.2018 18.01.2018
Principal growth stage 6: Flowering
61: Beginning of flowering: 10% of flower hoods fallen
67: 70% of flowerhoods fallen
Principal growth stage 7: Development of fruits
71: Fruit set: young fruits begin to swell, remains of flowers lost
77: Berries beginning to touch (if bunch are tight)
I observation (31.05.2017) 70% of flowerhoods fallen; II observation (10.06.2017)
Small-berry grape only formats; III observation (01.07.2017) Berries beginning to touch
Muscat Hamburg ‘Muscat d’Adda’ ‘Afuz Ali’ ‘Victoria’
Isolation of the inflorescences Isolation of the inflorescences Isolation of the Isolation of the inflorescences
+ pinching fertile shoots + pinching fertile shoots inflorescences + pinching + pinching fertile shoots
‘Muscat Hamburg’ (a) ‘Muscat d’Adda’ (a) fertile shoots ‘Afuz Ali’ (a) ‘Victoria’ (a)
Pollination open Pollination open Pollination open Pollination open
‘Muscat Hamburg’ (b) ‘Muscat d’Adda’ (b) ‘Afuz Ali’ (b) ‘Victoria’ (b)
283
Figure 3. Evolution of the rainfall (%) Figure 4b. Evolution of fruit set (b)
for the period 20 may-20 june 2017
Figures 5a, 5b provide details of the size
27th of May 2017 for all experimental variants, categories in which the berries are engraved,
in order to observe the percentage of berries expressed as a percentage, and it was noticed
formed (fruit set), self-pollination and open that ‘Muscat Hamburg’ had developed very
pollination, on one hand, and the uniformity of high percentage of normal berries (76.1%),
their size, on the other hand, it was observed followed by the ‘Muscat d’Adda’ (74%) and
that the percentage of berries binding (fruit set) ‘Afuz Ali’ (71.1%), and the lowest percentage
varies from one species to another (Figure 4a), of normal berries was registered by ‘Victoria’
as follows: ‘Muscat d’Adda’ varieties and variety with only 55.5%, when the two cultural
‘Muscat Hamburg’ recorded the lowest techniques were applied.
percentage (21% and 23%, respectively),
followed by ‘Afuz Ali’ (31%) and the highest
percentage of berries formed by the ‘Victoria’
variety (48%), in the case of self-pollination
and pinching fertile (a).
Figure 5a. Evolution of the categories

of grains formed (a)
For experimental variants with open

pollination, the same things have been noticed,
Figure 4a. Evolution of fruit set (a) but with slightly higher values. Interesting and
also surprising is that although ‘Afuz Ali’ and
In the case of open pollination, a similar
‘Victoria’, for the two experimental variants,
evolution had been observed, but the had the highest percentage of berries formed
percentages registered a higher value for all (fruit set), the proportion of grapes with a very
four grape varieties (Figure 4b). small berries (2-4 mm millet grain size) and
This shows that some varieties, even if they small berries (6-7 mm pea size) was more
have normal functional hermaphrodite flowers, pronounced. Thus, the ‘Victoria’ variety had
they have proved to have incompatibility with recorded 44% non-uniform berries in the case
their own pollen (Lepadatu, 1979) as seen in of the isolation of the inflorescences and 42%
the ‘Afuz Ali’ variety. in the case of open pollination.
284
reaches at least 16 0Brix. Grapes with a lower
refractometric index are accepted provided the
sugar/acid ratio (Total Soluble Solids/Tritatable
Acidity – TSS/TA) is at least equal to 20:1 if
the Brix level is comprised between 12.5 ° and
14 °Brix, 18:1 if the Brix level is comprised
between 14° and 16 0Brix. Some table grapes,
like the varieties chosen in this study can
accumulate TSS higher than 16 °Brix, with low
levels of acidity, thus leading to a TSS/AT ratio
even greater than 30:1, (Antonacci et al., 2017).
Some table grapes, as in the present case, can
accumulate TSS higher than 16 ° Brix with low
levels of acidity, leading to a TSS/AT ratio
Figure 5b. Evolution of the categories of berries formed (b)
even greater than 30: 1.The concentrated values
of sugars are followed by an acidity that gives
Variety ‘Afuz Ali’ had recorded smaller values,
the varieties balanced sugar-acidity indices, the
but different in both studied variants, a
highest values being recorded in ‘Afuz Ali’ (b)
precentage of 29% in the isolation of the
5.52, and the lowest value for ‘Victoria’ 3.61,
inflorescence and 34% in open pollination. This
‘Victoria’ variant (a).
things indicates that it had been influenced by
the climatic conditions on the last days of
CONCLUSIONS
flowering phenophase when temperature
recorded was 16ºC. The results obtained at this
The special cultural techniques for table grape
stage of development (flowering) are outlined
varieties have a positive impact on quantity and
in the quantitative and qualitative parameters of
quality parameters, but all of them on the
the obtained products (Table 2), reaching a
background of a set of generous climatic
high productive potential due to the large grape
factors, recorded in critical phenophase of vine
sizes, whose values are within the performance
(flowering, ripening). In all experimental
limits of the varieties analyzed. Regarding the
variants, the quantities of sugar accumulated
quality, which is appreciated by the size of the
had been adequate, being in correlation with
berries and their uniformity, the data show at
acidity, ensuring in this balanced sugar-acidity
least for the ‘Victoria’ variety that the
indices and also in line with the standards
remaining berries are large, uniform in size,
demanded by the consumer in the market.
especially for grapes of the ‘Victoria’ variant
(a) were the applied technology resulted in
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Table 2. Evolution of quality parameters on the experimental varieties
0 Total
Brix Gluco- Average Average
Experimental acidity Yield
(as Total Soluble acidometric weight of a weight of a
Varieties (g tartaric (kg/vine)
Solids, g/L) index berries (g) grape (g)
acid/L)
12.68 3.51 3.61 6.44 889.47 9.33
a
109.87
‘Victoria’
14.12 3.63 3.89 5.0 673.1 7.06
b
124.74
18.66 5.04 3.70 2.96 460.42 14.04
a
‘Muscat 172.59
Hamburg’ 22.13 4.65 4.76 3.34 364.06 8.00
b
209.7
19.22 5.3 3.63 3.97 502.23 11.80
a
Muscat 178.9
d.Adda 19.6 5.1 3.84 4.61 433.34 6.39
b
183.15
18.26 3.67 4.98 4.20 432.65 5.98
a
168.28
‘Afuz Ali’
20.7 3.75 5.52 5.55 447.88 6.19
b
194.83
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INFLUENCES OF GRAPE CULTIVATION TECHNOLOGY ON CERTAIN

AROMA COMPOUNDS SIGNIFICANT FOR THE DIFFERENTIATION OF
‘CABERNET SAUVIGNON’ AND ‘FETEASCĂ NEAGRĂ’ WINES
George Adrian COJOCARU1, Arina Oana ANTOCE1*

1
59 Marasti Ave., District 1, 011464, Bucharest, Romania
*Corresponding author emails: [email protected]; [email protected]
Abstract
Red wine grape varieties ‘Fetească neagră’ and ‘Cabernet Sauvignon’ were produced in Murfatlar wine region under
organic and conventional viticultural systems. For both varieties and viticultural systems, 30% cluster thinning was
performed in some variants during summer, at the beginning of veraison. The resulted grapes, harvested in the autumn
of 2014, were vinified using the same winemaking protocol and the differences in aromatic profile of the wine samples
were assessed by using a GC e-nose from Alpha MOS. The chromatographic peaks were recorded and analysed with
Alpha Soft ver. 12.42 and AroChemBase. The discrimination of the samples was based on multivariate DFA statistical
analysis. The results showed that 30% cluster thinning did not influence significantly the major wine aroma
compounds, only small differences being observed in the DFA diagram for samples with and without cluster thinning.
However, the grape variety and the application of organic cultivation technology led to a good discrimination, with a
validation score of 92, of the samples on the DFA diagram. An even better discrimination, with a validation score of 96,
was observed on wine samples from grapes produced in conventional and organic system when the cluster thinning was
disregarded and samples with or without this intervention in the vineyard were included in the same group. These
results show that, as far as the aromatic profile is concerned, the wines produced from grapes to which cluster thinning
was applied are difficult to differentiate from the controls produced from vines with normal yield, even with a sensitive
equipment such as an e-nose. On the other hand, the organic and conventional growing led to wine samples easily
differentiated by the electronic nose, suggesting that these differences can also be perceived by consumers. Some of the
aroma compounds significant for the discrimination of wines produced from organic or conventional grapes were
identified and their expected sensory effect is discussed.
Key words: ‘Fetească neagră’, ‘Cabernet Sauvignon’, electronic nose, aroma profile, organic viticulture, cluster
thinning.
INTRODUCTION favouring aroma maturation and lowering the

vegetative growth, which in turn favours the
The global quality of wine aroma is well formation of flavours (Jackson, 2008). On the
related with grape-growing conditions and other hand, an increased potassium content in
healthy grapes, along with the winemaking soil may lead to an increase in C13
technology involved, being an important norisoprenoids in grapes and of certain acetate
attribute for consumers buying decisions and a esters produced during fermentation in wines
challenge for the winemakers. The primary (Falcao et al., 2008). Type of soil and drainage
aroma of wines is derived from grapes and show an influence on the aroma composition of
implicitly from cultivation technology, along grapes and in the resulted wines, such as
with climate and soil variables, while the metoxypyrazines decrease in grapes on gravelly
secondary aroma is related to the winemaking soil (Ribéreau-Gayon et al., 2006), more floral,
processing techniques and enzymatic sweet and fruity aromas develop in wines
transformations of grape compounds by derived from clay soils, solvent-like and green
microorganisms, especially during alcoholic notes appear in wines derived from sandy soils
fermentation. Viticultural practices and (Gómez-Míguez et al., 2007) while more
ecosystem factors influence the expression of terpenoids, sesquiterpenoids, and C13
certain genes involved in production of aroma norisoprenoids are found in wines derived from
compounds and precursors in grapes. Limited calcareous soils (Coelho et al., 2009).
soil fertility influences the ripening process However, the type of soil and drainage, directly
287
influence the soil temperature which appear to esters was observed (Webster et al., 1993).
influence aroma compounds in wines with the Plant protection with fungicides may lead to
highest accumulation of β-damascenone and residues on the grapes that affect the yeasts and
geraniol in warmer soils and conversely, a fermentative performance which in turn
lower accumulation β-damascenone but higher influence aroma quality (Noguerol-Pato et al.,
accumulation in β-ionone in cold soils. Dry 2011; Gonzales-Rodriguez et al., 2011;
weather at the end of summer improves fruit Gonzalez-Alvarez et al., 2012a and 2012b).
quality through the advance of ripening and
favours the flavour formation (Jackson, 2008), MATERIALS AND METHODS
while the temperature influences the
development of C6 aldehydes in cool regions The grapes used for this study were from the
and higher concentrations of monoterpenes in ‘Cabernet Sauvignon’ and ‘Fetească neagră’
warmer regions (Ji and Dami, 2008). Excessive cultivars, vintage 2014, cultivated in Murfatlar
sunlight exposure or partially shading of grape wine region. The vine parcels used in this study
clusters modify the terpenol contents. In are located close to each other, in order to
Muscat cultivars, the highest concentrations of reduce the variability induced by soil and
terpenols were found in artificially 50% shade climate. The organic cultivation technology for
(Belancic et al., 1997), while a 90% shade both varieties are subject to actual regulations
decreased the terpenols and C13 norisoprenoids and controlled by Council Regulation (EEC)
(Bureau et al., 2000a, 2000b). The 834/2007. An additional viticultural practice
concentration of TDN (1,1,6-trimethyl-1,2- applied consisted in 30% cluster thinning
dihydronaphthalene) increased with sunlight during summer, at the beginning of veraison,
exposure in Riesling (Gerdes et al., 2002), being evaluated against the control with no
while in the other study on Shyrah, an extreme cluster thinning. The grapes were harvested on
shading lead to a decrease in concentration of 16th September for the ‘Fetească neagră’ and on
TDN (Ristic et al., 2007). Viticultural practices 24th September for ‘Cabernet Sauvignon’. In
on different varieties regarding cluster thinning this manner eight experimental variants were
and basal leaf removal showed an increase of produced, according with viticultural practices
free terpenols and glycosidically bond terpenols briefly described in Table 1. More details
(Reynolds et al., 1996b; Roberts et al., 2007; regarding the grapes physico-chemical
Hernandez-Orte et al., 2014; González-Barreiro composition and other grape related parameters
et al., 2015). However, different training are described in other paper (Artem et al., 2015).
systems studied by various authors showed The grape processing was done for batches of
variations in terpenols concentrations (Ji and grapes of 50 kg, in 3 repetitions resulting 24
Dami, 2008; Reynolds et al., 1996a; Zoecklein samples. The vinification process was the classical
et al., 2008). Water stress may increase one, with maceration and fermentation on skins
blackberry, jam, raisin or dried fruit aroma in for 5-8 days for ‘Fetească neagră’ and for 8-10
‘Cabernet Sauvignon’ wines (Chapman et al., days for ‘Cabernet Sauvignon’. The
2005), increase of cysteinylated thiol fermentation was conducted at room
precursors in grapes of ‘Sauvignon blanc’ temperature at about 22-24°C with native
(Ribéreau-Gayon et al., 2006), increase of yeasts (no added yeast), without added
glycoconjugates of the main aromatic nutrients or enzymes. At the end of
compounds observed in Agiorgitiko vines fermentation the macerated grape marc of each
(Koundouras et al., 2006), increase of variant was pressed using a laboratory press.
vitispirane, guaiacol, 4-methylguaiacol, 4- After one month of clarification and cold
ethylguaiacol and 4-vinylguaiacol observed in stabilization, the resulted wines were racked off
Merlot vines (Qian et al., 2009). Fertilization of and treated with a dose of 50 mg/L of SO2 to
vineyard with nitrogen lead to higher prevent oxidation and development of spoilage
cysteinylated thiol precursors in ‘Sauvignon microorganisms. The resulted wine samples
blanc’ grapes (Choné et al., 2006), while in were analysed in other 3 repetitions on 7th May
Riesling wines a rise in 1-butanol, trans-3- 2015, assessed with a GC e-nose from Alpha
hexen-1-ol, benzyl alcohol and the majority of MOS in order to establish the differences
288
among certain aroma compounds between having a different polarity, one being non-polar
experimental variants. DB5 (5% diphenyl, 95% dimethylpolysiloxane)
Alpha MOS Heracles e-nose analyser is a Fast and the other with a low/mid polarity DB1701
GC using a Tenax trap for pre-concentration of (14% cyanopropylphenyl, 86%
volatile organic compounds, designed with two dimethylpolysiloxane).
short capillary columns working together
Table 1. Experimental variants and the related codes attributed
Variety Grape cultivation technology Additional practice Variant code

‘Cabernet Sauvignon’ Conventional No cluster thinning CS_Con_N
30% Cluster thinning CS_Con_CT
Organic No cluster thinning CS_Org_N
30% Cluster thinning CS_Org_CT
‘Fetească neagră’ Conventional No cluster thinning FN_Con_N
30% Cluster thinning FN_Con_CT
Organic No cluster thinning FN_Org_N
30% Cluster thinning FN_Org_CT
The separated volatile organic compounds are main volatile organic compounds identified to
detected by means of simultaneous dual flame differentiate wine samples in this study are
ionization detectors (FID), located at the end of briefly described in Table 2. The results show
each chromatographic column. The injection small differences between the experimental
mode was made with 2.5 ml HS syringe, variants with 30% cluster thinning as compared
extraction of volatiles from the head-space of to no cluster thinning variants, this being
vials, using a 250°C injector temperature with especially evident for conventional ‘Fetească
an initial column temperature 40°C with an neagră’ (Figure 1). This clearly led to the idea
increase rate of a 5°C/s up to 200°C and that cluster thinning practice did not
hydrogen in constant pressure (16 psi) as a significantly influence the major volatile
carrier gas. The oven agitator temperature for organic compounds in wine samples and only
sample preparation was 10 minutes at 60°C and small changes being observed, which are hardly
500 rpm. The flame ionization detectors recognized even by a powerful tool such as
temperature have been set at 220°C and a fuel GC-e-nose. However a good validation score of
pressure of 35 psi. Other details regarding the 92 in DFA diagram (Figure 1) is explained by
methodology and Alpha MOS Heracles can be the higher variability induced by both grape
found in other papers (Antoce et al., 2015; cultivation technology (organic or
Antoce and Namolosanu, 2011). The conventional) and variety. According to Figure
identification of peaks was based on retention 1, the DFA bi-plot explained 96.554% of the
index (Kovats), calibrated by using a known total variance in the data through the first two
standard n-alkane mixture (n-C6 to n-C16) with dimensions, with 85.069% and 11.485%
the same determination conditions as for the explained by DF1 (mostly grape variety
analysed wine samples. The discriminant factor related) and DF2, respectively. In general in
analysis (DFA) was selected in Alpha Soft v Figure 1 and 4, no cluster thinning, appear to be
12.42 software as the most relevant statistical associated with more negative volatile organic
method to separate the experimental variants compounds as 2,4-hexadienal, 2,3-octanedione
and groups. and valeric acid, while 30% cluster thinning,
appear to be associated with more positive
RESULTS AND DISCUSSIONS volatile organic compounds as 3-methyl-1-
pentanol, dehydro-p-cymene, β-phenylethanol,
In order to differentiate wine samples, nerol oxide and (E)-2-hexenal. The results
discriminant factor analysis was applied to regarding cluster thinning practice did not show
certain chromatographic peaks area (sensors), a clear difference between those samples as can
selected by using an extensive iterative process be seen in Figure 4, where DFA bi-plot have a
and their discriminant power provided by validation score of 69.
Alpha Soft v 12.42. Using this process, the
289
Figure 1. DFA bi-plot of aroma compounds peaks used for differentiation of wine sample groups in accordance to grape
variety, grape cultivation technology (organic/conventional) and cluster thinning practice
Table 2. Relevant aroma compounds identified as being discriminative in wine samples

of ‘Cabernet Sauvignon’ and ‘Fetească neagră’
*Retention Column *Sample Database Sensor Identified volatile Sensory descriptor

time Kovats Kovats number organic compounds
8.05 DB5 844,04 843 844,04-1-A 3-methyl-1-pentanol chocolate, wine-like, green
10.36 DB5 909,32 908 909,32-1-A 2,4-hexadienal green, citrus, floral
12.54 DB5 962,54 962 962,54-1-A γ-valerolactone anise, herbal, sweet, warm, tabacco, cocoa,
woody, hay-like, coumarinic odor,
17.72 DB5 1090,66 1094 1090,66-1-A dehydro-p-cymene citrus, pine
18.70 DB5 1117,07 1117 1117,07-1-A β-phenylethanol sweet, floral, fresh, bready, rose, honey
11.25 DB1701 953,38 956 953,38-2-A (E)-2-Hexenal sweet, vegetable, almond, apple, green, plum
14.34 DB1701 1028,93 1031 1028,93-2-A valeric acid cheese, sweat
16.10 DB1701 1072,03 1073 1072,03-2-A γ-valerolactone anise, herbal, sweet, warm, tabacco, cocoa,
woody, hay-like, coumarinic odor,
16.49 DB1701 1081,23 1082 1081,23-2-A 2,3-octanedione dill, cooke, broccoli, buttery
19.58 DB1701 1156,97 1157 1156,97-2-A dehydro-p-cymene citrus, pine
27.26 DB1701 1356,93 1353 1356,93-2-A nerol oxide flower, oil
*average values resulted from all chromatograms;
Nevertheless, the cultivation technology and functions, with DF1=76.516% and

variety are the most important variables in this DF2=20.336%. From the loadings of the
study, for which reason we have redefined the discriminant peaks also included in Figure 2 it
groups according to only cultivation technology can be seen that conventional ‘Fetească
and variety, removing the cluster thinning neagră’ is associated with more valeric acid
variable (Figure 2). Using this approach, a than the organically cultivated ‘Fetească
better validation score (96) is obtained and a neagră’, which is associated with 2,4-hexadienal,
very good separation on the DFA bi-plot while the conventional ‘Cabernet Sauvignon’ is
(Figure 2). In Figure 2, the DFA bi-plot associated with β-phenylethanol and organic
explained 96.852% of the total variance in the ‘Cabernet Sauvignon’ with nerol oxide,
data through the first two discriminant dehydro-p-cymene and (E)-2-hexenal.
290
Figure 2. DFA bi-plot of aroma compounds peaks used for differentiation of wine samples depending on grape variety
and grape cultivation technology (organic/conventional)
Figure 3. DFA bi-plot of aroma compounds peaks used for differentiation of wine samples depending on grape
cultivation technology (organic/conventional)
When we have redefined the groups according oxide, dehydro-p-cymene, 3-methyl-1-

to only cultivation technology removing the pentanol, 2,4-hexadienal and (E)-2-hexenal and
variety and cluster thinning variables (Figure slightly with 2,3-octanedione. However, the
3), all organic samples are well differentiated organic and conventional growing led to wine
by conventional samples with a good validation samples easily differentiated by the electronic
score of 96 observed in the DFA bi-plot. In nose, suggesting that these differences can also
figure 3 we can observe the main differences be perceived by consumers. In one study of
between organic and conventional samples and sensory evaluation of red wines deriving from
the DFA bi-plot explained 100% of the total organically and conventionally grown grapes,
variance in the data through the first dimension. trained assessors and regular wine consumers
The results from the figure 3 reveal that attributed the differences mainly to sour and
conventional growing technology in this study bitter taste, with astringency sensations, while
is associated with more β-phenylethanol, γ- aroma have a minor role (Pagliarini et al.,
valerolactone and valeric acid, while the organic 2013).
grape production is associated with more nerol
291
Figure 4. DFA bi-plot of aroma compounds peaks used for differentiation of wine samples depending on cluster
thinning
CONCLUSIONS wine as the pure substances are perceived and

described in literature, as their mixture in the
Grape variety followed by the type of grape wine matrix changes very much this perception.
technology (organic/conventional) were the However, these findings serve as hints to
most discriminative factors for the wines winemaker to find ways to improve the quality
produced and evaluated in this study, allowing of grapes and wines.
for a good separation of wine samples in the
DFA bi-plots, with high validation scores (96). ACKNOWLEDGEMENTS
This means that the grape growing technology
has a significant impact on grape aroma The authors wish to express their gratitude to
compounds and implicitly in wines, meaning the Research Station for Viticulture and
that these modifications are probably perceived Oenology Murfatlar, which kindly provided the
even by regular wine consumers. The 30% wine samples analysed in this study with the
cluster thinning did not influence significantly electronic nose. The wines were produced at
these aroma compounds compared with the the Research Station for Viticulture and
samples with no cluster thinning and in this Oenology Murfatlar from grapes grown in their
case, certainly, discrimination of wine samples own organic and conventional experimental
by the electronic nose could not be achieved in blocks.
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294
INCREASE OF ANTHOCYANINS ACCUMULATION

BY PRE-HARVEST 24-EPIBRASSINOLID (24-EBL) APPLICATIONS
IN ʻHOROZ KARASIʼ GRAPE CULTIVAR
Zehra BABALIK1, Tunhan DEMİRCİ2, Özlem ARAS AŞCI2, Nilgün GÖKTÜRK BAYDAR2
1
Suleyman Demirel University, Atabey Vocational School, 32670, Atabey-Isparta-Turkey
2
Suleyman Demirel University, Agricultural Faculty, Agricultural Biotechnology, 32260, Isparta-
Turkey
Abstract
In this study, the effect of pre-harvest 24-eBL (a BR analogue) applications on the accumulation of anthocyanin of
ʻHoroz Karasiʼ grape cultivar was examined. 24-eBL was applied to vines with 13 combinations including different
application times (veraison, 7 days after berry set + veraison, 7 days after berry set + veraison + 30 days after
veraison) and 4 different concentrations of 24-eBL (0.2, 0.4, 0.6 and 0.8 mg/L) and control. As a result of the study, the
highest anthocyanin content was obtained from the vines applied with 0.2 mg/L of 24-eBL at 7 days after fruit set and at
veraison stages in both years (respectively, 47.98 and 71.16 mg/100 g FW).
Key words: anthocyanin, brassinosteroid, Grape 24-eBL, pre-harvest application.
INTRODUCTION Brassinosteroids (BRs), known as the sixth

group of hormones, have highly beneficial
Grapes are one of the most consumed fruits, effects on plant growth, development and
and due to its rich phenolic compounds, such as physiological aspects in plants such as seed
antiradical and antioxidant properties, interest germination, rhizogenesis, flowering, rooting,
in grapes and grape products (especially food senescence, abscission, cell expansion and
additives, pharmaceutical industry and natural elongation and they are considered as plant
cosmetic products) is increasing (Bourgaud et hormones with having pleiotropic effects
al., 2001; Ghafoor et al., 2009). (Clouse and Sasse, 1998; Vardhini and Rao,
Anthocyanins in grapes have well-known many 2002; Luan et al., 2013).
beneficial effects for human health including Moreover exogenous applications of BRs
the reduction in the incidence of coronary heart increase the synthesis of secondary metabolites
disease, enhancement of visual acuity, in plants (Biesaga-Koscielniak et al., 2014;
maintenance of normal vascular activity, as Ghorbani et al., 2017).
well as pharmacological properties and strong Symons et al. (2006) and Luan et al. (2013)
biological functions such as antiinflammatory showed that the exogenous application of BR to
and antioxidant activities (Hohnova et al., grape berries evidently enhanced skin
2008). Also, anthocyanins in grapes play an coloration and anthocyanin accumulation.
important role in the quality of colour and they Although there are some studies showing that
are successfully used as food colourants and BRs influence the expression of anthocyanin
nutraceuticals (Espín et al., 2007). Therefore, biosynthesis genes in grapes (Luan et al., 2013;
the determination of anthocyanins in red grapes Xi et al., 2013), little is known about the
has acquired of increasing interest during last possible roles of BRs involved in anthocyanin
decade. Many different applications can be accumulation. In this study we investigated the
made in cultivation in order to enrich the effect of BRs on anthocyanin accumulation in
nutrient and antioxidant contents of grapes. The grapes.
most important treatments in grape growing for Thus, the objective of this study was to
this purpose are the usage of plant growth determine the effect of pre-harvest 24-eBL
regulators. application at different doses and application
295
times on anthocyanin accumulation in ʻHoroz RESULTS AND DISCUSSIONS
Karasiʼ grape cultivars.
Effects of 24-eBL concentrations and its appli-
MATERIALS AND METHODS cation periods on anthocyanin contents were
given in Table 1. 24-eBL applications generally
The experimental vineyard was located in increased the anthocyanin accumulation. In the
Senirkent-Isparta Province in the both years, interaction effects of 24-eBL
Mediterranean region of Turkey. Nine-year old concentrations and application periods on the
Vitis vinifera L. cv. ʻHoroz Karasiʼ grapevines anthocyanin contents were statistically
grafted 41 B.M.G. were used. The vines were significant (p˂0.05).
planted at 2 x 3 m spacing trained on a bilateral In the first year, the highest anthocyanin
cordon system. 24-eBL was sprayed during two content was observed at 7 days after fruit set
years at concentrations of 0, 0.2, 0.4, 0.6 and and at veraison periods with 0.2 mg/L 24-eBL
0.8 mg/L and prepared by dissolving in DMSO. (47.98 mg/100 g FW) and at 7 days after fruit
Tween 20 at the rate of 0.1% (v/v), as a set + at veraison periods with 0.6 mg/L 24-eBL
surfactant was added to 24-eBL solution. The (45.35 mg/100 g FW) and at 7 days after fruit
prepared solutions were sprayed directly onto set + at veraison + at 30 days after verasion
the vine (1L per vine) at veraison periods with 0.6 mg/L 24-eBL (45.35 mg/100 g
(approximately 10% of the berries of 50% of FW).
the clusters become soft and at colour break), at In the second year, the treated with 0.2 mg/L
7 days after fruit set (3-5 mm diameter berry 24-eBL at 7 days after fruit set and at veraison
size) + at veraison and 7 days after fruit set and periods grapes had the highest anthocyanin
at veraison + 30 days after veraison by a hand contents. The lowest anthocyanin contents were
pump sprayer until run-off early in the observed in control grapes for both years.
morning. Grapes were harvested at commercial
Table 1. Anthocyanin contents (mg/100 g FW) in ʻHoroz
maturity and bunches were packed in plastic Karasiʼ grapes as effected by 24-eBL concentrations and
crates and transported to the laboratory where application periods
they were frozen in liquid nitrogen and stored Application Concentrations
at -80°C until the analyses were performed. period (AP) (C) (mg/L)
1st year 2nd year
Total anthocyanin content determination was 0 29.36 g 39.73 e

0.2 33.38 f 41.24 de
based on a pH differential method and Veraison (V) 0.4 36.40 d 57.14 abcd
expressed as malvidin 3-glucoside equivalents 0.6 35.22 e 62.34 abc
0.8 22.73 j 51.46 bcde
(Wrostad, 1976). For this purpose, aliquots of 0 29.38 g 39.73 e
the extracts were adjusted to pH 1.0 and 4.5 7 days after 0.2 47.98 a 71.16 a
fruit set +
with buffers. The absorbance of each solution veraison
0.4 21.19 l 42.23 de
0.6 45.35 b 65.35 ab
was measured at wavelength of 520 and 700 (FS+V)
0.8 29.41 g 47.33 cde
nm. 7 days after 0 29.38 g 39.73 e
The experiment was conducted in randomized fruit set + 0.2 27.50 h 42.78 de
veraison + 30 0.4 22.40 k 52.02 bcde
complete block design with 3 (application time; days after 0.6 39.75 c 41.08 de
at veraison, at 7 days after fruit set + at veraison 0.8 23.86 l 60.83 abc
(FS+V+30V)
veraison and 7 days after fruit set + at veraison Main effect (Stages)
+ 30 days after veraison) 5 (24-eBL concen- V 31.42 50.38
FS+V 34.66 53.16
tration; 0, 0.2, 0.4, 0.6 and 0.8 mg/L) 2 FS+V+V30 28.58 47.29
(years; first and second) factorial arrangements. Main effect (Concentrations)
Each treatment had three replications of eight 0 29.38 39.73
0.2 36.29 51.73
vines. 0.4 26.66 50.46
Statistical analyses were performed with LSD 0.6 40.11 56.26
0.8 25.33 53.21
multiple range test. p values
Differences were considered statistically AP ˂0.0001 0.2991
C ˂0.0001 0.0197
significant at the p≤0.05 levels. The software AP x C ˂0.0001 0.0035
JMP 8 (SAS Institute, Inc., Cary, NC) was used Different letters indicate significant differences between groups
for carrying out statistical analyses of the data. (p<0.05).
296
Also, the main effect of 24-eBL application REFERENCES
periods and concentrations on anthocyanin con-
tents were shown in Table 1, too. In both years, Biesaga-Koscielniak J., Dziurka M., Ostrowska A.,
when the main effects of 24-eBL application Mirek M., Koscielniak J., Janeczko A., 2014.
Brassinosteroid Improves Content of Antioxidants in
periods and concentrations examined, the Seeds of Selected Leguminous Plants. Australian
application at 7 days after fruit set + at veraison Journal of Crop Science, 8 (3): 378-388.
periods and 0.6 mg/L 24-eBL concentration Bourgaud F., Gravot A., Milesi S., Gontier E., 2001.
had the highest anthocyanin contents, respec- Production of plant secondary metabolites: a
tively. Anthocyanins are synthesized by the historical perspective. Plant Science, 161, 839-851.
Champa W.H., Gill M.I.S., Mahajan B.V.C., Aror N.K.,
biosynthetic way regulated by enzymes such as Bedi S., 2015. Brassinosteroids improve quality of
PAL, chalcone synthase (CHS), chalcone table grapes (Vitis vinifera L.) cv. Flame Seedless,
isomerase (CHI), F3H, DFR, leucoantho- Tropical Agricultural Research, 26 (2): 368-379.
cyanidin dioxygenase (LDOX) (UFGT). The Clouse S.D., Sasse J.M., 1998. Brassinosteroids:
rise in these enzymes causes the synthesis and Essential Regulators of Plant Growth and
Development. Annual Review of Plant Physiology
accumulation of anthocyanin. BRs can increase and Plant Molecular Biology, 49, 427-451.
the amount of anthocyanin as a result of the Espín J.C., García-Conesa M.T., Tomás-Barberán F.A.,
enhancement in the enzymes on this synthetic 2007. Nutraceuticals: Facts and fiction.
route, and increase in the expressions of the Phytochemistry, 30, 2986-3008.
genes encoding these enzymes. Indeed, it was Ghafoor K., Choi Y.H., Jeon J.Y., Jo I.H., 2009.
Optimization of ultrasound-assisted extraction of
determined that BRs can stimulate the expres- phenolic compounds, antioxidants, and anthocyanins
sions of the genes involved in the synthesis of from grape (Vitis vinifera) seeds. Journal of
anthocyanin, thus increasing the amount of agricultural and food chemistry, 57 (11): 4988-4994.
anthocyanin in grapes (Xi et al., 2013). Peng et Ghorbani P., Eshghi S., Haghi H., 2017. Effects of
al. (2011) reported that BRs affect the brassinosteroid (24-epibrassinolide) on yield and
quality of grape (Vitis vinifera L.) Thompson
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jasmonates, through the regulation of late Hohnova B., Štavíková L., Karásek P., 2008.
anthocyanin biosynthetic genes such as BRL, Determination of anthocyanins in red grape skin by
DFR, LDOX and UFGT. Similarly, previous pressurised fluid extraction and HPLC. Czech J.
studies showed that pre-harvest BRs applica- Food Science, 26, 39-42.
Luan L.Y., Zhang Z.W., Xi Z.M., Huo S.S., Ma L.N.,
tions increase the amount of total anthocyanin 2013. Brassinosteroids regulate anthocyanin
compared to control application in grapes, too biosynthesis in the ripening of grape berries. South
(Luan et al., 2013; Xi et al., 2013; Champa et African Journal of Enology and Viticulture, 34 (2):
al., 2015). 196-203.
Peng Z., Han C., Yuan L., Zhang K., Huang H., Ren C.,
2011. Brassinostreoid enhances jasmonate-induced
CONCLUSIONS anthocyanin accumulation in Arabidopsis seedlings.
Journal of Integrative Plant Biology, 53 (8): 632-640.
As a result of investigations, it was seen that all Symons G.M., Davies C., Shavrukov Y., Dry I.B., Reid
pre-harvest 24-eBL applications increased the J.B., Thomas M.R., 2006. Grapes on steroids,
amount of anthocyanin compared to the control brassinosteroids are involved in grape berry ripening.
application. Accordingly, it is revealed that Plant Physiology, 140, 150-158.
among the 24-eBL applications, the better Vardhini B.V., Rao S.S.R., 1998. Effect of
brassinosteroids on growth, metabolite content and
results were obtained from 0.2 mg/L 24-eBL yield of Arachis hypogaea, Phytochemistry, 48 (6):
applicated at 7 days after fruit set and at 927-930.
veraison periods. So, this application could be Wrostad R.E., 1976. Color and pigment analysis in fruit
recommended for ʻHoroz Karasiʼ cultivar. products. Station Bulletin 624, Agricultural
Experiment Station Oregon State University,
Corvallis.
ACKNOWLEDGEMENTS Xi Z.M., Zhang Z.W., Huo S.S., Luan L.Y., Gao X., Ma
L.N., Fang Y.L., 2013. Regulating the secondary
This research work was carried out with the metabolism in grape berry using exogenous 24-
support of TUBITAK (Scientific and epibrassinolide for enhanced phenolics content and
Technological Research Council of Turkey) antioxidant capacity, Food Chemistry, 141 (3): 3056-
and also was financed from Project (Project 3065.
number: TOVAG 112O517).
297
298
THE INFLUENCE OF PRE-HARVEST BORIC ACID APPLICATIONS

ON THE ACCUMULATION OF SOME ANTIOXIDANT COMPONENTS
IN ALPHONSE LAVALLÉE GRAPE CULTIVAR
Zehra BABALIK1, Tunhan DEMİRCİ2, Özlem ARAS AŞCI2, Nilgün GÖKTÜRK BAYDAR2
1
Suleyman Demirel University, Atabey Vocational School, 32670, Atabey-Isparta,Turkey
2
Suleyman Demirel University, Agricultural Faculty, Agricultural Biotechnology,
32260, Isparta-Turkey
Abstract
The aim of the research was carried out to determine the effects of boric acid (BA) applications on the accumulation of
anthocyanins, β-carotene, ascorbic acid and total phenolics in ʻAlphonse Lavalléeʼ grape cultivar. BA was applied to
vines with four different concentrations (250, 500, 750 and 1000 mg/L) and at two different periods (at blooming and at
blooming+fruit set). At the end of the study, it was determined that the BA applications consist of 250mg/L and 750
mg/L of boric acid concentrations during both blooming and fruit set stages were the most effective applications providing
the highest anthocyanins, β-carotene and total phenolics. On the other hand, boric acid applications had no important
effects on the ascorbic acid contents in berries.
Key words: ʻAlphonse Lavalléeʼ, antioxidant components, boric acid, pre-harvest applications.
INTRODUCTION methyl jasmonate, ozone) and invoking biotic

and abiotic stresses have been used in both
Antioxidants are substances that, at relatively plant cell culture and intact plants (Dong et al.,
low concentrations, acting to scavenge and 2010; Krzyzanowska et al., 2012). However,
stabilize of free radicals. The benefits of optimizing the agricultural practices also
antioxidants are very important to good health. provide a rapid and efficient way to increase
Antioxidants can protect the human body free the antioxidant content. Mineral nutrients are
radicals that may cause some chronic diseases essential for the growth, survival and
including cancer, cardiovascular diseases and reproductive success of plants. Among the
cataract (Shahidi, 2000; Pham-Huy et al., essential elements for healthy plants, boron (B)
2008). Consumers are becoming increasingly is responsible for activation of dehydrogenase
prefer aware antioxidant substances rich in enzymes, sugar translocation, nucleic acids,
fruits and vegetables because of their beneficial plant hormones and effect on cell wall structure
effects on health. Grape is one of the most cell elongation, root growth and transfer of
important fruit commodities as widely grown sugar and altered production of a wide range of
through the world. Also, it is a significant phenol content and metabolism. In addition to
source of nutritional antioxidants, such as it is helpful in growth and productivity, fruit
polyphenols, anthocyanins as well as setting and yield. B deficiency has visual
biologically active dietary components (Orak, symptom on root and leaves growth, flower,
2007). Therefore, due to its rich phenolic cluster and berry development in grapevine.
compounds, such as antiradical and antioxidant Severely reduced in internodes and shoot
properties, interest in grapes and grape products length, shoot tip death, low fruit set, and tiny
(especially food additives, pharmaceutical berries are all common symptoms of B
industry and natural cosmetic products) is deficiency (Fleischer et al., 1998; Ruiz et al.,
increasing (Bourgaud et al., 2001; Ghafoor et 1998; Ebadive et al., 2001; Brown et al., 2002;
al., 2009). To enhance the production of Christensen et al., 2006; Singh et al., 2012).
antioxidant substances, strategies such as Several studies have demonstrated that B
treating with elicitors (UV, salicylic acid, application increased the yield in grapes
299
(Mostafa et al., 2006; Er et al., 2011; Ally et triplicate. β-carotene content of the sample was
al., 2015; Asci et al., 2017). However, although measured using the method of Association of
there is limited information on the effect of B Official Analytical Chemists (AOAC, 1980).
on the antioxidant components of plants and The absorbance of the extracts was measured at
there is no information about grapes. Therefore, 436 nm wavelength by using a
the major objective of this study was to spectrophotometer. The concentration of β-
determine the effect pre-harvest B application carotene was calculated using Bear-Lamberts
on the accumulation of some antioxidant Law. Total anthocyanin content determination
components (total phenolics, ascorbic acid, β- was based on a pH differential method and
carotene, anthocyanins) in ʻAlphonse Lavalléeʼ expressed as malvidin 3-glucoside equivalents
grape cultivars. (Wrolstad, 1976). For this purpose, aliquots of
the extracts were adjusted to pH 1.0 and 4.5
MATERIALS AND METHODS with buffers. The absorbance of each solution
was measured at wavelength of 520 and 700
The experiment was performed on 9-year old nm. The experiment was conducted in 2
Vitis vinifera L. cv. ʻAlphonse Lavalléeʼ (application time; at blooming, at blooming +
grafted on 41 B.M.G. planted at 2 x 3 m at fruit set) 5 (BA concentration; 0, 250, 500,
spacing trained on a bilateral cordon system in 750 and 1000 mg/L) factorial arrangements
a commercial vineyard in the Senirkent-Isparta with completely randomized block design.
Province in the Mediterranean region of Turkey Each treatment had three replications of 8
(lat. 38º 11' 8'' N, long. 30º 40' 55'' E and vines. Statistical analyses were performed with
elevation 981 m). Eight vine at similar LSD multiple range test. Differences were
conditions were used. Boric acid (BA) was considered statistically significant at the p≤0.05
sprayed at concentrations of 0, 250, 500, 750 levels. Statistical analyses were performed by
and 1000 mg/L. Tween 20 at the rate of 0.1% using JMP 8 (SAS Institute, Inc., Cary, NC).
(v/v), as a surfactant was added to BA solution.
The prepared solutions were sprayed directly RESULTS AND DISCUSSIONS
onto the vine (1 L per vine) at blooming and at
blooming + fruit set stages by a hand pump In concerning with influences of BA spraying
sprayer until run-off early in the morning. on antioxidant properties of Alphonse Lavallée
Grapes were harvested at commercial maturity grape, data was shown in Table 1 and Table 2.
and bunches were packed in plastic crates and Data showed that the interaction effects of BA
transported to the laboratory where they were concentrations and application periods had a
frozen in liquid nitrogen and stored at -80°C significant (p≤0.0001) influence on total
until the analyses were performed. Total phenolic, β-carotene and anthocyanin contents.
phenolic content was estimated by the Folin- Grapes tretaed with 250 and 750 mg/L BA at
Ciocalteu colorimetric method, based on the blooming and at fruit set stages produced more
procedure of Singleton and Rossi (1965), using total phenolic compound (16.84 mg/g and
gallic acid as a standard phenolic compound. 16.32 mg/g, respectively) compared to control
The reduction of the Folin-Ciocalteu reagent by and the other applications. Similarly, the
phenolic compounds under alkali conditions, highest anthocyanin content was observed with
which resulted in the development of a blue 250 and 750 mg/L BA at blooming and at fruit
colour, was measured at 765 nm using a set stage, too (66.89 and 66.55 mg/100 g)
spectrophotometer. The total phenolic content (Table 1).
was calculated with the use of a calibration As shown Table 2, β-carotene content was also
curve and results were expressed as mg gallic higher 250 mg/L BA at blooming and at fruit
acid equivalent per g dry weight. L-Ascorbic set stage (698.32 µg/kg) followed by 750 mg/L
acid content was determined BA at blooming and at fruit set stage (422.75
spectrophotometrically at 520 nm using a µg/kg). Data showed that there was no
standart curve according to the procedure significant difference observed among the
described by Pearson and Churchill (1970). treatments for ascorbic acid content. Ascorbic
Each determination was carried out in acid content varied from 1.97 to 2.73 mg/100 g.
300
Table 1. Response of BA spraying at different stage on total phenolic (mg/g DW)
and anthocyanins (mg/100 g FW) of ʻAlphonse Lavalléeʼ
Concentrations (C) Total phenolics Anthocyanins
Application period (AP)
(mg/L) (mg/g DW) (mg/100 g FW)
0 7.59 f 35.24 de
250 11.33 e 46.67 b
Blooming (B) 500 14.40 b 30.33 e
750 12.13 cde 42.03 bc
1000 13.03 c 35.78 cde
0 7.59 f 35.24 de
250 16.84 a 66.89 a
Blooming + Fruit set (B+FS) 500 12.73 cd 41.60 bcd
750 16.32 a 69.55 a
1000 11.88 de 37.39 cd
Main effect (Stages)
B 11.70 38.01
B + FS 13.07 50.13
Main effect (Concentrations)
0 7.59 35.24
250 14.09 56.78
500 13.56 35.96
750 14.22 55.79
1000 12.46 36.59
p values
AP ˂0.0001 ˂0.0001
C ˂0.0001 ˂0.0001
AP x C ˂0.0001 ˂0.0001
Different letters indicate significant differences between groups (p<0.05).
Also, the main effect of BA application periods processes such as carbohydrate transport,
and concentrations on antioxidant properties which are enhanced by its application
were shown in Table 1, too. When the main (Marschner, 2012; Davarpanah et al., 2016).
effects of BA application periods and Due to the fact sucrose is a positive regulator of
concentrations examined, they were obtained the biosynthesis of phenolics, the improvement
that at blooming + at fruit set periods and 250 of B treatment on photosynthesis and sugar
mg/L BA concentrations had the highest total accumulation could possibly enhance the
phenolics, ascorbic acid, anthocyanins and β- biosynthesis of phenolics (Solfanelli et al.,
carotene contents (Tables 1 and 2). 2006). To our knowledge, there is very little
studies examining the relationship between
phenolics and B. There were no comprehensive
wine, and their contents are and detailed studies about the effect of sprayed
considered as the most important nutritional B on the whole development process included
value parameters. B is one of the nutrients antioxidant properties of grapes. But some
responsible for the changes in concentration investigators have observed a positive effect of
and metabolism of phenolic compounds in B on total phenolic contents in fruits. For
vascular plants. It is well known that B instance, foliar sprays of B increased total
deficiency causes an accumulation of phenolics phenolic compounds in olive (Saadati et al.,
through the stimulation of the enzyme 2013) and in pungent pepper (Manas et al.,
phenylalanine-ammonium lyase (PAL) 2014). The authors indicate that the increase
(Cakmak et al., 1995). B is rapidly absorbed by was due to an increased expression of genes
flowers (Sarrwy et al., 2012) and the B responsible to phenolic compound
application effects can be attributed to its effect biosynthesis.
on fruit set and development or other metabolic
301
Table 2. Response of BA spraying at different stage on ascorbic acid (mg/100 g FW)
and β-carotene (µg/kg FW) of ʻAlphonse Lavalléeʼ
Application period Concentrations (C) Ascorbic acid β-carotene
(AP) (mg/L) (mg/100 g FW) (µg/kg FW)
0 1.97 289.65 bcd
250 2.60 321.04 bcd
Blooming (B) 500 2.62 352.34 bcd
750 2.69 281.56 cd
1000 1.79 392.61 bc
0 1.97 289.65 bcd
250 2.73 698.32 a
Blooming + Fruit set
500 2.19 273.21 cd
(B+FS)
750 2.52 422.75 b
1000 2.25 245.90 d
Main effect (Stages)
B 2.33 327.44
B + FS 2.33 385.96
Main effect (Concentrations)
0 1.97 289.65
250 2.66 509.68
500 2.41 312.78
750 2.60 352.15
1000 2.02 319.25
p values
AP 0.9965 0.0626
C 0.1000 0.0010
AP x C 0.6670 0.0001
Different letters indicate significant differences between groups (p<0.05).
Ascorbic acid acts as an antioxidant in plants. The lack or additional of B may also influence
According to some researchers its levels are efflux of other macro and micro elements
responsive to a variety of environmental or (Camacho-Cristobal et al., 2008; Hajiboland
stress factors such as light, temperature, salt and Farhanghi, 2010). Singh et al. (2012) found
and drought, pollution, metals or herbicides that the accumulation of P, K, Mg, S, Na, Al
(Hancock and Viola, 2005; Nicolle et al., and Mn in the carrot storage roots increased
2004). Lukaszewski and Blevins (1996) when additional B was not supplied to the
reported that adequate level of B is required for plants. This alteration of mineral levels could
the accumulation of ascorbic acid. Although either directly or indirectly affect the
there was no significant difference observed accumulation of phytonutrients, such β-
among the B treatments for ascorbic acid carotenes.
content in our investigation, Mondi and Munsi
(1993) reported that applied B significantly CONCLUSIONS
increased the ascorbic acid concentration in
potatoes. According to the study, since B plays Previous studies were mostly concerned the
an important role in the translocation of effect of B on the quality of grapes but, there
carbohydrates from leaves to other portions of were no comprehensive and detailed studies
the plant, greater concentrations of ascorbic about the effect of sprayed B on the whole
acid may have been translocated to the tuber. development process included antioxidant
Also, Govindan (1950) reported that the properties of grapes. Obtained results are
ascorbic acid concentration of tomatoes rose important to increase the antioxidant properties
with increased B uptake in the plant. Cakmak of ʻAlphonse Lavalléeʼ grapes. As a result of
and Romheld (1997) have shown that the investigations, the better results were obtained
decline in vitamin C in some plant parts such as with 250 mg/L BA applicated at blooming and
shoot tips and young leaves of sunflower under at fruit set periods and B application can be
B deficient conditions. recommended for ʻAlphonse Lavalléeʼ grape
It is known that B is important for the structural cultivar.
and functional integrity of plasma membranes.
302
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304
MOLECULAR DETECTION OF BLACK ASPERGILLUS AND

PENICILLIUM SPECIES FROM DEALU MARE VINEYARD
Camelia Filofteia DIGUȚĂ, Radu Cristian TOMA, Călina Petruța CORNEA,

Florentina MATEI
University of Agronomic Sciences and Veterinary Medicine of Bucharest, Faculty of

Biotechnologies, 59 Mărăşti Blvd, District 1, Bucharest, Romania
Abstract
The aim of this study was to determine the incidence and diversity of black Aspergillus and Penicillium species on white
and red grape varieties in Dealu Mare vineyard (in two vilticultural centres - Valea Calugareasca and Pietroasa),
during the harvest time of 2014 and 2015. A total of 61 fungal strains belonging to black Aspergillus (28 strains) and
Penicillium (33 strains) were isolated. An RFLP analysis of the 5.8 S-ITS region was performed by using different
combinations of endonucleases for molecular identification of fungal isolates at species level. A. niger (accounting
60.71% of all isolates) and P. expansum (66.66%) were the predominant species identified, followed by A. tubingensis
(17.85%) and P. chrysogenum (21.21%). Higher biodiversity of fungal isolates has been found in Pietroasa Centre than
from Valea Calugareasca.
Key words: black Aspergillus, Penicillium sp., PCR-ITS RFLP, grapes, Dealu Mare vineyard.
INTRODUCTION Also, several species of Aspergillus section

Nigri and Penicillium are recognized as
Filamentous fungi (or molds) are communally potentially producers of mycotoxins
contaminants of grapes. In recent years, (ochratoxin A, patulin etc.) and volatile
particular attention has been drawn of black compounds (geosmin, 2-methyl-isoborneol
Aspergillus and Penicillium on grapes, being etc.) (La Guerche et al., 2004; Garcia et al.,
considered the main spoilage agents of grapes 2006; Martinez-Culebras and Ramon, 2007;
and grape-derived products and is responsible Somma et al., 2012; Rousseaux et al., 2014).
for significant economic losses. Rousseaux et Among the black aspergilli, A. carbonarius has
al. (2014) reviewed thirty-six species of been considered as the main OTA producer
Aspergillus and fifty-nine different species of (Belli et al., 2006), followed by species
Penicillium identified on grapes in vineyards belonging A. niger aggregate (A. niger and A.
around the world. tubingensis) (Bejaoui et al., 2006; Perrone et
The incidence of the species of black al., 2006). Among Penicillium species, P.
Aspergillus and Penicillium has been studied expansum is the major causal agents of the blue
on grape varieties from vineyards worldwide: mold rot of grapes and the main source of
France (La Guerche et al., 2004; Garcia et al., volatile compounds production which
2006; Bejaoui et al., 2006; Diguță et al., 2011; determinate off-flavours (La Guerche et al.,
Ahmed et al., 2015); Portugal (Serra et al., 2004, Garcia et al., 2006).
2006); Spain (Martínez-Culebras and Ramón, The detection of mycotoxins and volatile
2007; Sardinas et al., 2011; Sempere et al., compounds in grape wines can influence not
2011; Garcia-Cela et al., 2014); Italy (Ayoub et only detrimental to the sanitary quality, but also
al., 2010; De Rossi et al., 2011; Spadaro et al., to the organoleptic quality (La Guerche et al.,
2012; Somma et al., 2012); Greece (Kizis et al., 2004; Garcia et al., 2006; Martinez-Culebras
2014; Kogkaki et al., 2015); Slovakia and Ramon, 2007; Somma et al., 2012;
(Felsociova et al., 2013; Santini et al., 2014; Rousseaux et al., 2014). In Romania, Geana et
Felsociova et al., 2015); Romania (Diguță et al. (2012) reported the detection to traceable
al., 2011; Diguță et al., 2015; Diguță et al., OTA levels (ranging from 0.06 ng/mL to 0.45
2016). ng/mL) in 7 wine commercialized on the
305
Romanian market, however, the concentrations by Diguță et al. (2011). The suspension was
found being far below the proposed European serially diluted and dilutions were plated on
limit (2 ng/mL). Black aspergilli and Dichloran Rose-Bengale Agar (Mecconti,
Pencillium species are difficult to taxonomical Poland) supplemented with Chloramphenicol.
identification by classical methods. Among Plates were incubated at 25°C in the dark for 5-
molecular approaches, restriction fragment 7 days. Only fungal strains considered to
length polymorphism (RFLP) based on ITS- represent black Aspergillus and Penicillium
5.8S region have been proved useful tool to the species were isolated and maintained to Potato
identification and classification the species dextrose agar (PDA, Mecconti, Poland). The
belonging to Aspergillus section Nigri (Accensi pure cultures were preliminary screened by
et al., 1999; Bau et al., 2006; Martínez- colony characteristics (mycelium type, color
Culebras and Ramón, 2007; Kizis et al., 2014); and growth type) as well as microscopic
to differentiate P. expansum among fungal characteristics (conidiophores and spore) (Pitt
grape species (Garcia et al., 2006); to and Hocking, 2009; Varga et al., 2011).
discriminate 22 species belonging to
Penicillium genus and 7 species belonging to Table 1. Grape samples used in this study
Aspergillus (Diguţă et al., 2011). Vilticultural Grape variety Acronym
The latest available data of The International centres
Organisation of Vine and Wine (OIV) shows Valea Fetească Regală (white grape) FRV
that Romania is considered in top viticultural Călugărească Cabernet Sauvignon (red grape) CSV
production potential with an area under vines Fetească Regală Conventional FRCP
the 191 thousands of hectares of overall in (white grape)
Cabernet Sauvignon CSCONP
European vineyards (OIV, 2017a). After two Conventional (red grape)
poor harvests, Romania returned to a high level Pietroasa
Cabernet Sauvignon Ecologic* CEP
of production (5.3 millions of hectoliters) (red grape)
(OIV, 2017b). Tămâioasă Românească TăRCP
Consequently, there is a special need to manage Conventional (white grape)
and combat the spoilage molds directly in the *The vineyard is under certified ecological conditions.
vineyard in order to obtain high quality of the
Romanian wines which can compete with high DNA extraction
quoted wines coming from other traditional Fungal isolates were grown in 10 ml of PDB
wine-making countries (France, Italy, Spain Broth (Mecconti, Poland) at 25ºC for 72 h.
etc.) or more recent producers as USA, Mycelia were collected after centrifugation
Argentina or South Africa. (10000 rpm for 5 minutes) and washed with
In this context, this study was focused to sterile saline water then frozen at -25°C until
determinate the incidence and diversity of use. Approximately 200mg of fungal biomass
black Aspergillus and Penicillium species on was used for DNA extraction by the use of ZR
white and red grape varieties in Dealu Mare Fungal/Bacterial MiniPrep™ Kit (Zymo
vineyard (in two vilticultural centres - Valea Research, USA), according to the
Calugareasca and Pietroasa), during the harvest manufacturer’s instructions.
time of 2014 and 2015.
PCR-RFLP conditions
MATERIALS AND METHODS Amplification of the 5.8S-ITS region was
performed with universal primers ITS1 (5'-
Fungal isolation TCCGTAGGTGAACCTGCGG-3', forward) or
The grape samples (white and red grape ITS5 (5’-GGAAGTAAAAGTCGTAACAAGG-
varieties) were randomly and aseptically 3’, forward) and ITS4 (5’-
collected from Dealu Mare vineyard (in two TCCTCCGCTTATTGATATGC-3’, reverse) in
vilticultural centres - Valea Calugareasca and order to identify fungal strains (White et al.,
Pietroasa) at harvest time in 2014 and 2015 1990). The PCR program for the amplification
season (Table 1). Fungal isolation was with ITS1/ITS4 was as follows: 2 min at 94°C,
performed according to the method described 34 cycles with 1 min at 94°C, 1 min at 55°C, 2
306
min at 72°C, with a final extension for 7 min at electrophoresis, respectively. Restriction
72°C. The PCR program for the amplification fragments smaller than 70bp could not be
with ITS5/ITS4 was as follows: 2 min at 95°C, clearly visualized and were not included in our
35 cycles consisting of 1 min at 95°C, 1 min at results. Restriction patterns were compared
52°C and 1 min at 72°C, with a final extension with RFLP patterns obtained by Martínez-
for 10 min at 72°C. Culebras and Ramón (2007) and Diguţă et al.
PCR products were digested with different (2011).
combination of restriction enzymes Cfr9I,
Hpy188I, HinfI, HhaI, MseI, NlaIII, RsaI and RESULTS AND DISCUSSIONS
SduI (Thermo Scientific, USA), according to
PCR-ITS-RFLP methods developed by A total of 61 fungal strains including 28 strains
Martínez-Culebras and Ramón (2007) and black Aspergillus and 33 strains Penicillium
Diguţă et al. (2011) (Figure 1). were isolated in this study. All the isolates have
been taxonomically identified by molecular
methods using PCR-RFLP. Twenty eight
fungal strains belonging to Aspergillus section
Nigri have been isolated. The primer pairs
ITS5/ITS4 were used to amplify the 5.8S-ITS
region, followed by restriction enzyme
digestion with three endonucleases HhaI,
NlaIII and RsaI, according to ITS-RFLP
method developed by Martínez-Culebras and
Ramón (2007) (Table 2). Based on RFLP
types, four species belonging to black
Aspergillus have been identified: A. niger
aggregate (A. niger and A. tubingensis), A.
aculeatus, A. japonicus (Table 2).
A. niger (17 of all the isolates) was dominated
among isolated fungus from black aspergilli,
followed by A. tubingensis (5 isolates).
Moreover, two strains belonging to the
uniseriate species Aspergillus aculeatus and A.
japonicus showed the same PCR-RFLP.
Figure 1. Identification of black Aspergillus and Digestion with HinfI showed two distinct
Penicillium isolates by PCR-RFLP based 5.8S-ITS restriction patterns: 110+180+270 (for A.
region, adapted after Diguţă et al. (2011) (A) and aculeatus) and 270+290 (for A. japonicus)
Martinez-Culebras and Ramón (2007) (B); M - (personal data). Based on RFLP profile, 4
GeneRuler 100bp Plus DNA Ladder isolates have been identified as A. japonicus.
Thirty-three fungal strains belonging to
PCR- RFLP analysis Penicillium genus have been isolated.
The amplification and digestion results have Amplification of the 5.8S-ITS region was
been visualized under UV (254 nm) after performed with the universal primers
electrophoresis in agarose gel 2% (90V/60 ITS1/ITS4 followed by digestion with the
minutes). All fragment sizes were evaluated by combination of three endonucleases SduI, HinfI
the fluorescence intensity and were and MseI to discriminate Penicillium isolates at
approximated using the DNA ladder species level, according to PCR-ITS-RFLP
(GeneRuler 100bp Plus DNA Ladder, Thermo method developed by Diguta et al. (2011)
Scientific, USA). Restriction fragments are (Table 3).
considered to be distinguished when the According to Table 3, of the Penicillium
difference between them is more than 20bp and isolates, 22 isolates showed a RFLP profile
considered to be fragments in common if corresponding to P. expansum. Seven isolates
migrated the same distance during agarose gel have been identified as P. chrysogenum. This
307
result has been confirmed with other management and combating of these molds
endonucleases Cfr9I or Hpy188I to differentiate directly from the vineyard. However, the
P. chrysogenum from P. crustosum and P. impacts of climate changes and viticultural
commune (personal data). In addition, other 4 practices on the occurrence of black
isolates of Penicillium sp. showed new RFLP Aspergillus and Penicillium on grapes have
profiles. To validate our results, sequencing is been not taken into account, when have been
required. analysed the results obtained in this study.
The distribution of identified species on grapes According to figure 2 we found that A. niger
varieties in Dealu Mare vineyard is given in was the predominant species isolated from
Figure 2. Higher biodiversity of fungal isolates grapes. A. carbonarius, the main species of
has been found in Pietroasa Centre than from ochratoxigenic species, have not been isolated.
Valea Calugareasca (Figure 2 A and B). However, in another study, A. carbonarius has
The distribution of Aspergillus sp. and been detected and quantified by qPCR in all
Penicillium sp. on the grape varieties can vary naturally grape samples tacking in account in
from one vineyard to another, from one year to this study (Diguță et al. 2016).
another, grape maturity, climatic conditions and
viticultural practices being key factors, which
leads to difficulty in generalizing the
Table 2. RFLP analysis of 5.8S-ITS region exhibited by the Aspergillus isolates
No. Aspergillus sp. Number Restriction fragments
isolates HhaI (pb) NlaIII (pb) RsaI (pb)
1. A. niger 17 90 120 180 210 110+130+360 520
2. A. tubingensis 5 90 120 180 210 110 130 360 80+480
3. A. aculeatus 1 70 140 180 180 220+350 80+480
4. A. japonicus 4 70 140 180 180 220 +350 80 +480
Table 3. RFLP analysis of 5.8S-ITS region exhibited by the Penicillium isolates

No. Penicillium sp. Number Restriction fragments
isolates SduI (pb) HinfI (pb) MseI(pb)
1. P. expansum 22 170 270 110 180 290 110 110 360
2. P. chrysogenum 7 170 270 290 290 110 110 360
3. Penicillium 1 1 170 270 110 180 290 560
4. Penicillium 2 1 170 270 290 290 560
5. Penicillium 3 1 190 320 290 290 110 110 360
6. Penicillium 4 1 560 130 160 290 190 240
On the Fetească Regală grape variety (in 2014) varieties (Fetească Regală and Tămâioasă
and Cabernet Sauvignon (in 2015), no strain of Românească) and international grape variety
Aspergillus sp. or Penicillium sp. have been Cabernet Sauvignon from Pietroasa Centre
isolated (Figure 2A). Other species commonly (Figure 2B). Diversity of Aspergillus sp. and
found on grapes, namely Alternaria alternata, Penicillium sp. is higher from Tămâioasă
Cladoporium cladosporioides, Epicoccum Românească variety (TăRCP - for 2014) and
nigrum on both varieties were identified Fetească Regală variety (FRCP - for 2014 and
(personal data). A certain fungal diversity was 2015) from Pietroasa Centre (Figure 2 B). P.
observed on Fetească Regală grape variety (in expansum was detected on FRCP (both years
2015), predominantly P. expansum, followed season), TăRCP (2014) and CSCONP (2015).
by other new Penicillium species, A. Also, P. chrysogenum was another highly
tubingensis (Figure 2A). isolated species on the two grape varieties
A special attention has been given to fungal analyzed (TăRCP, for 2014 and FRCP, for
mycobiota of two autochthonous grape 2015). A. niger was isolated predominantly on
308
the Cabernet Sauvignon Conventional Variety Two other uniseriate Aspergillus species, A.
(CSCONP) in 2014 season. Also, A. niger was japonicus and A. aculeatus were punctually
identified on the Cabernet Sauvignon isolated on CSCONP (both seasons), TăRCP
Ecological (CSEP) variety in 2015. A. (2014) and FRCP (2015) (Figure 2B).
tubingensis was predominated CSEP (2014)
and TăRCP (2015) (Figure 2B).
Figure 2. Distribution of black Aspergillus and Penicillium isolates in two vilticultural centres -
Valea Călugărească (A) and Pietroasa Centre (B) on different grape varieties
CONCLUSIONS ACKNOWLEDGEMENTS
A total of 61 fungal isolates have been isolated We would like to acknowledge the help from
from red and white wines cultivated in Dealu dr. Elena Brîndușe (Valea Călugărească) and
Mare vineyard under conventional and dr. Sorin Marin (Pietroasa) for all samples
ecological conditions. collected.
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310
INFLORESCENCE AND FLOWER DIFFERENTIATION IN THE

GRAPEVINE (V. VINIFERA L.) VARIETIES ʻŽILAVKA’ AND ʻBLATINA’
Tatjana JOVANOVIC-CVETKOVIC1, Nikola MICIC1, 2,

Gordana ĐURIĆ1, 2, Miljan CVETKOVIĆ1
1
Faculty of Agriculture, University of Banja Luka
2
Genetic Resources Institute, University of Banja Luka
Bulevar vojvode Petra Bojovića 1A, 78000 Banja Luka, Bosnia and Herzegovina
Abstract
The different dynamics of flower differentiation i.e. flowering and the functional capacity of reproductive organs in
grapevine must be studied as genotypic specificity under particular agroenvironmental conditions and brought to a
level at which standardized biological control of fruiting is exercised in order to define sustainable production systems.
Research on the functional morphology of the inflorescence and flower in the indigenous grapevine varieties 'Žilavka'
and 'Blatina' as the major ones used for grape production in Bosnia and Herzegovina (B&H) was conducted in 2010-
2011 through analysis of permanent histological slides. The morphological characteristics of the inflorescence, as
observed through the presence and distribution of flowers, and the flowering dynamics of individual flowers in the
inflorescence pave the way for the normal course of flowering and pollination in the studied varieties. During
inflorescence development, the degree of flower congregation in the distal part of the inflorescence is defined as a
varietal characteristic. The inflorescence in 'Žilavka' and 'Blatina' grapevines forms a cone-shaped panicle due to the
different structure of side branches i.e. different morphology and structure of branching on the axes of simple botryose
inflorescences along the inflorescence axis, when observed from the top down to the base. The sequence and dynamics
of differentiation of the flower generative elements clearly indicate protandry in both varieties.
Key words: grapevine, inflorescence, flower, characteristic
INTRODUCTION producing functionally female flowers that

have a large effect on yield, which varies
The grapevine inflorescence is a panicle depending on the year. The production of a
belonging to compound racemose single-variety wine from this variety has not
inflorescences with the axis growing forward been a regular winemaking practice. As the
relatively long at the tip forming side branches 'Blatina' varietal wine does not exhibit long-
with flowers arranged in simple racemose lasting color stability, it is recommended to
inflorescences, and terminating in a flower blend wines made from varieties used as
(closed inflorescence). The grapevine pollenizers in plantings (Blesić, 2001). On the
inflorescence differentiates in the bud from the other hand, 'Žilavka' has been the leading
end of May until September in a single growing grapevine variety in B&H for many years now
season (Mulins et al., 1992; Burić, 1981) and (Mijatović, 1988; Tarailo, 1991). As estimated
continues to develop in the spring of the by Tarailo and Kovačina (1983), the important
following year. As stressed by Carmona et al. role 'Žilavka' plays in grapevine production and
(2008), this fact is the main constraint to many its heterogeneity in cluster and berry
studies aiming to explore all differentiation appearance make it possible to select a range of
sequences and finally execute the genetic genotypes. However, when analyzing the
control of grapevine reproduction and yield. A genetic variability of 'Žilavka' by the AFLP
more detailed analysis of flower differentiation technique Tomić (2009) determined the genetic
in the inflorescence in accordance with the profile characteristic of 'Žilavka' at 14
classification proposed by Mulins et al. (1992) microsatellite loci and found no genetic
has been provided by Caporali et al. (2003). background for the selection of 'Žilavka' clones
'Blatina' is an indigenous grapevine variety or new genotypes with improved varietal
311
properties. The objective of this study was to inflorescence is defined as a varietal charac-
examine the functional morphology of the teristic. This aggregation and grouping of
inflorescence and flower in 'Žilavka' and flowers in simple botryose inflorescences on
'Blatina' grapevines as the basis of success in side branches in the basal half of the panicle is
micro- and macrosporogenesis. the result of the progressive shortening of the
internodes on the side branches of the
MATERIALS AND METHODS inflorescence. Reduction in internode length on
side branches which carry simple botryose
Anatomical, morphological and cytohisto- inflorescences is a variety-specific morpho-
logical characteristics of reproductive organs logical character in ampelography as it results
were evaluated in two major indigenous in variable compactness of the infructescence
varieties in B&H - 'Blatina' and 'Žilavka'. Part (the arrangement of berries in the cluster), and
of the experimental research and sampling for at preflowering it can be associated with the
the anatomical, morphological and mechanical pressure on petals, that is, this
cytohistological analyses of reproductive compactness among flowers can have a role
organs were conducted in a commercial towards flower caps in terms of flower
vineyard of the Aluminium Plant, Mostar opening-cap fall.
(southeastern B&H). The varieties were grafted
on Kober 5BB rootstock, with vines trained to
the Moser cordon system and subjected to short
pruning. Standard agricultural and viticultural
practices were used, which along with growing
conditions (modified Mediterranean climate)
ensured steady yields. Samples for the
preparation of histological slides were collected
from initial cluster emergence until
fertilization. The cytohistological analysis of
the reproductive organs involved preparation of
permanent histological slides. Sampling for the
preparation of histological slides was
performed during shoot and inflorescence
growth and at early flowering and pollination
stages. Inflorescences were sampled every two
days between 9 and 10:30 a. m. from 27 April
until 25 May in 2009 and 2010. In the field,
immediately after removal from the shoots, the
inflorescences were placed into bottles and Figure 1. Structural analysis of side branches of an
fixed in Navashin's fixative. The fixed paraffin- inflorescence histologically (left) and morphologically
embedded specimens were sectioned using a (right) showing flowers grouped in simple clusters in the
following arrangement - the apical part of the side branch
microtome at a thickness of 9 ƞm. The terminating in three flowers which have bract rudiments
differential staining of the histological sections at their base; remaining flowers grouped in simple
was performed by Delafield’s hematoxylin. botryose - racemose inflorescences whose number and
The sections were analyzed under an distribution on the shortened to compact axes of the side
Olympus/DP/SOFT light microscope. Images branches of the inflorescence can be observed primarily
through bract presence
were documented using an Olympus/DP
camera and edited by the Image Analyzer
software. The morphological characteristics of the
inflorescence cannot be directly associated with
RESULTS AND DISCUSSIONS functional morphology as the presence and
distribution of flowers and the flowering dyna-
During inflorescence development, the degree mics of individual flowers in the inflorescence
of flower congregation in the distal part of the cannot be associated with pollination or fruit
set. The first branch of the inflorescence closest
312
to the base is often quite distant from the other
branches and constitutes a distinct part of the
inflorescence. During further growth and
development, this branch of the inflorescence
can develop into a small cluster called a lateral
cluster or a wing, which can reach the length of
the main inflorescence in some varieties. This
inflorescence character has been established in
'Žilavka' grapevine and is considered its
varietal characteristic as regards the
ampelographic characterization of the cluster
(Tarailo and Kovačina, 1983; Mijatović, 1988;
Tarailo, 1991). The differentiation of flower
primordia was observed dynamically from the
microstage at which the emergence of
inflorescence axes on the shoots was visible
until fully formed flowers (Carmona et al.,
2008). The research performed at this level of
observation showed no anatomical, a.
morphological or histological differences in the
differentiation of individual flowers in the
studied varieties. The difference in the
differentiation dynamics between the varieties
and among the inflorescences differently
positioned on the shoots at this point of the
research was analytically generalized so that
more extensive histological and cytological
analyses of individual flowers could be
executed for the evaluation of their individual
elements. The flower of the grapevine is
pentagonal, composed of 5 sepals arranged
along the margin of the receptacle. As the
sepals are poorly developed, they can be
considered rudimentary. The inside of the calyx b.
holds five petals which are fused along their
entire length. There are 5 stamens, completely Figure 2. Microstages of flower differentiation in
free and practically located in the axils of the 'Blatina': a) sepals, petals and anther primordia
differentiated on the flower axis; b) microstages of the
petals. Each stamen consists of an anther differentiation dynamics of flower elements observed in
supported by a filament. The pistil occupies relation to the sepals whose development is arrested as
the center of the flower, and comprises the early as the beginning of flower primordium
ovary, stigma and style. The ovary is bilocular, development, due to which they practically remain
having two carpels, morphologically two rudimentary. The petals fuse at the tips and cover anther
initials, keeping them leaning against the stigma of the
chambers, each containing two ovules. The ovary
histological analysis of flower primordia
becoming visible as a small pinhead shows the The differentiation sequence and dynamics of
differentiation of sepal primordia enclosing the the flower generative elements clearly indicate
less conspicuous petal primordia, which form a that both varieties are protandrous. Protandry is
roof over dome-shaped anther initials (Figure fully visible in the histological sections of the
2a). At the microstage when the petals surpass anthers and ovaries in the same flower initials
the sepals by their whole length, all anther (Figure 3).
primordia elements differentiate, and initial
carpel differentiation is observed (Figure 2b).
313
a.
Figure 3. Microstages of flower differentiation in

'Blatina'. At this microstage of flower primordium
development, the final stages of microsporogenesis and
initial stages of macrosporogenesis are observed
The results on the dynamics of flower

differentiation in the inflorescences of 'Žilavka'
and 'Blatina' grapevines, as observed on the
histological slides, are in complete agreement
with the flower differentiation dynamics
reported by Caporali et al. (2003) based on the
classification proposed by Mulins et al. (1992). b.
The cross-section through the center of the
flower shows the position of the anthers Figure 4. Histological cross-sections of flower initials in
'Žilavka' (a) and 'Blatina' (b) clearly showing the petal
relative to the petals and the manner in which fusion zone
the petals fuse to form the flower cap in the
studied varieties. Minor differences are
The partition between the chambers is formed
observed in the length of the fusion zone
by the bending and curving of the carpel
between the varieties (the fusion zone being
margins inwards towards the interior of the
somewhat longer in 'Žilavka' than in 'Blatina'),
ovary. In this way, the carpels fuse laterally,
which can affect the rate of cap detachment
dividing the ovary into two chambers. All
during bloom (Figure 4 a, b). The center of the
ovules are initiated perpendicular to the margin,
flower is occupied by the pistil made up of two
and then nucellar and integumentary cells grow
fused carpels - microsporophylls. The ovary is
and develop at different rates, giving rise to
bilocular - it consists of two chambers, each
anatropous ovules with the micropyle facing
containing two ovules.
the base of the ovary. In this way, four
anatropous ovules in marginal placentation are
formed in the ovary of each variety (Figures 5
and 6). The central part of the floral structure is
occupied by the ovary composed of two
chambers, each holding two ovules.
314
The analysis of inflorescence and flower
differentiation in the tested varieties is
consistent with the findings of Caporali et al.
(2003) who reported that functional differences
between the (male and female) flowers are
commonly recorded at this stage, when no style
and stigma formation occurs on the pistil of
male flowers. Due to these morphological
changes, the pistils of male and female flowers
are quite different at the next stage of
development, whereas their stamens still
remain very similar in appearance.
CONCLUSIONS
Figure 5. Histological representation of the cross-section The inflorescence in 'Žilavka' and 'Blatina'
of the bottom third of an individual flower, in 'Žilavka'.
The cross-section displays the formed ovules at the grapevines forms a cone-shaped panicle due to
integument differentiation stage. Cross-sections of the the different structure of side branches i.e.
filaments are visible in the zone between the ovary and different morphology and structure of
the petals. The cross-section of the filament in 'Žilavka' branching on the axes of simple botryose
clearly shows epidermal and parenchyma cells located at inflorescences along the inflorescence axis,
the periphery of the filament and somewhat smaller
central cells of the vascular tissue). This anatomical when observed from the top down to the base.
structure of the filament is typical of bisexual flowers in The degree of flower congregation in the distal
the grapevine part of the inflorescence can be defined as a
varietal characteristic. This aggregation and
grouping of flowers of simple botryose
inflorescences on side branches in the basal
half of the panicle is the result of the
progressive shortening of internodes on the side
branches of the inflorescence. The histological
analysis of the flower primordium visible as a
small pinhead at the morphological stage shows
differentiated primordia of sepals enclosing the
less conspicuous primordia of petals, which
form a roof over dome-shaped anther initials.
The differentiation sequence and dynamics of
the flower generative elements clearly show
that both varieties are protandrous. All
inflorescence and flower differentiation
processes in the tested varieties take their
normal course, with no anomalies potentially
adversely affecting pollination and fertilization
found at this level of observation.
ACKNOWLEDGEMENTS
Figure 6. Histological representation of the This research work has been supported by the
morphological and anatomical structure of a portion of Ministry of Science and Technology of the
the ovary in 'Žilavka'. The ovule develops in the interior Republic of Srpska under the projects
of the ovary and has marginal placentation. Later on, the “Anatomorphological and cytohistological
ovule develops into a seed. The cross-section shows the
ovule the size of a small bulge covered by the evaluation of reproductive organs of indigenous
integument grapevine cultivars in Bosnia and Herzegovina”
315
(Contract No. 06/6-020/961- 119/08) and Mijatović D., 1988. Ispitivanje karakteristika rodnosti i
“Genotypic characterization of male sterility in kvaliteta grožđa sorte Žilavka u interakciji važnijih
agroekoloških činilaca. Doktorska disertacija.
cv. 'Blatina'” (Contract No. 19/6- 020/961- Poljoprivredni fakultet Univerziteta u Sarajevo.
177/09). Mullins M.G., Bouquet A., Williams L.E., 1992.
Biology of the grapevine. Cambridge: Cambridge
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vina u zavisnosti od uslova vođjenja maceracije privredno-tehnološkim karakteristikama. Doktorska
kljuka Blatine. Doktorska disertacija. Poljoprivredni disertacija. Poljoprivredni fakultet Univerziteta u
fakultet Univerziteta u Sarajevu. Sarajevo.
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A., 2003. The arrest of development of abortive vrijednosti nekih varijeteta sorte žilavka u
reproductive organs in the unisexual flower of Vitis mostarskom vinogorju. Zbornik radova IRI Mostar.
vinifera ssp. silvestris. Sexual Plant Reproduction 15, Tomić L., 2009. Genetic characterization of the
291-300. grapevine variety Žilavka (Vitis vinifera L.) with
Carmona M.J., ChaibJ., Martinez-Zapater J.M., Thomas DNA markers. M.Sc. Thesis. Ljubljana, University of
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Experimental Botany, Vol. 59, No.10, pp. 2579-2596.
316
DETERMINATION OF THE PHENOLIC COMPOUNDS,

ANTIOXIDANT AND ANTIRADICAL ACTIVITIES OF SENIRKENT
KARASI GRAPE CULTIVAR’S SKIN AND SEEDS
Filiz HALLAÇ TÜRK1, Emine Sema ÇETİN2, Zehra BABALIK3,

Nilgün GÖKTÜRK BAYDAR4
1
Suleyman Demirel University, Faculty of Agriculture, Department of Horticulture, 32260, Isparta,
Turkey
2
Bozok University, Faculty of Agriculture, Department of Horticulture, Yozgat, Turkey
3
Suleyman Demirel University, Atabey Vocational High School, Atabey, Isparta, Turkey
4
Suleyman Demirel University, Faculty of Agriculture, Department of Agricultural Biotechnology,
32260, Isparta, Turkey
Abstract
Purpose of this study was to determine total phenolic content, phenolic composition, antioxidant and antiradical
activities of ʻSenirkent Karasıʼ grape cultivar’s skin and seeds. While total phenolic contents of grape skin and seeds
were determined by Folin-Ciocalteu method spectrophotometrically expressing the results in terms of gallic acid
(GAE), phenolic composition was analyzed by HPLC (High Performance Liquid Chromatograph). Antioxidant
activities of the grape skin and seeds were evaluated by reducing powers whereas antiradical activities were examined
using DPPH (1-diphenyl-2- picrylhydrazyl). Results showed that total phenolic contents of seeds and skin were 52.32
and 1.89 mg g-1 GAE g-1 DM, respectively. Antiradical activities of seed and skin extracts (100 ppm) were 95.90 and
16.22 %, respectively. Reducing powers of seeds were 1.64 at 250 ppm, and were 2.39 at 1000 ppm whereas
antioxidant activities) of skins were 0.08 at 250 ppm; 0.31 at 1000 ppm. Results showed that skin had higher amount of
phenolic compounds than seeds and gallic acid, catechin, cafeic acid, syringic acid, resveratrol, quarcetin, kaempherol,
p-cumaric acid were present in skin whereas only gallic acid, catechin, epicatechin were present in seeds. Seeds had
the highest values of epicatechin (746.94 µg g-1) while skins had the highest values of syringic acid (17.01µg g-1) and
gallic acid (5.29 µg g-1).
Key words: phenolics, antioxidant activity, antiradical activity, ʻSenirkent Karasıʼ.
INTRODUCTION grape skin and seeds have a variety of

polyphenol contents, high antioxidant property
There is an increasing interest on grape, grape and contain flavonoids (catechin, epicatechin,
products and other parts of grapevine due to procyanidins, anthocyanins), phenolic acids
rich chemical compounds they have. Increasing (gallic acid, ellagic acid) and stilbenes
interest on natural antioxidants resulted in an (resveratrol and piceids) (Jayaparakasha et al.,
increase in number of research on improvement 2003; Negro et al., 2003; Yılmaz and Toledo,
and evaluation of natural products that are rich 2006). However different parts of grape have
in phenolic compounds. Phenolic compounds different content of these above mentioned
that have very high antioxidant and antiradical compounds.
properties are substances that have direct effect ʻSenirkent Karasiʼ grape cultivar is a local
on quality, that give resistance ability to grape grown in Isparta province that is mainly
diseases and have pharmacologic features used as wine, table and dried consumption
(Macheix et al., 1990; Clausen et al., 1992; grape.
Ayed et al., 1999; Yi et al., 2006). In addition, Thus, the purpose of this study was to
phenolic compounds reduce risk of cancer and determine the total phenolic content, phenolic
heart diseases and lead to low density composition, antioxidant and antiradical
lipoprotein (LDL) due to their high antioxidant activities of ʻSenirkent Karasiʼ grape cultivar’s
properties. There are studies showing that skin and seeds.
317
MATERIALS AND METHODS Diode Array Detector with wavelengths set at
278 nm. The 250 x 4.6 mm i.d. 5 µm column
Materials used was filled with Agilent Eclipse XDB-C18
In the study skin and seeds of ʻSenirkent (Wallborn, Germany). The flow rate was 0.8 ml
Karasiʼ, a commonly grown cultivar in Isparta, min-1, the injection volume was 20 µl, and the
was used. Fresh grapes were obtained from column temperature was set at 30°C. For
Isparta Directorate of Provincial Food gradient elution, mobile phase A contained 3%
Agriculture and Livestock, dried in the shade acetic acid in water; solvent B contained
and later seeds and skin were separated to be methanol. The following gradient was used: 0-
analyzed. There were three replications for 3 min, from 100% A to 95% A; 3-20 min, from
each analysis. 95% A to 80% A; 20-30 min, from 80% A to
75% A; 30-40 min, from 75% A to 70% A; 40-
Phenolic extraction 50 min 70% A to 60% A; 50-55 min, 60% A to
Grape seeds and skins were manually separated 50% AB; 55-65 min, 50% A to 0% A. The data
from whole berries, seeds were dried at room were integrated and analyzed using the
temperature and then were crushed in a grinder Shimadzu Class-VP Chromatography Labora-
for two min. In order to remove the fatty tory Automated Software system. The grape
materials from seeds, the powdered grape seeds samples, standard solutions and mobile phases
(100 g) were extracted in a Soxhlet extractor for were filtered by a 0.45 µm pore size membrane
6 h with 150 ml of petroleum ether at 60°C. filter (Millipore Co. Bedford, MA). The
The defatted grape seed powder and also amount of phenolic compounds in the seed and
powdered skin were extracted in a Soxhlet skin extracts were calculated as mg 100 g-1
apparatus for 8 h with 200 ml of acetone: extract, separately, using external calibration
water: acetic acid (90:9.5:0.5) at 60°C as curves obtained for each phenolic standard.
described by Jayaprakasha et al (2003). The Caffeic acid, (+)-catechin, chlorogenic acid, o-
extracts were concentrated by rotary evaporator coumaric acid, p-coumaric acid, (-)-
at 70°C to get crude extracts and stored in a epicatechin, ferulic acid, gallic acid, kaem-
desiccator. pherol, trans-resveratrol, quercetin, syringic
acid and vanillin acquired from Sigma (St.
Determination of total phenolic content Louis, MO, USA) were used as standards and
Total phenolic contents of the grape seed and determined in the samples.
skin extracts were determined spectrophoto-
metrically using a PG Instruments T70 Plus Determination of antiradical activity
Dual Beam Spectrophotometer (Arlington, The free radical scavenging activity of extracts
MA, USA) according to the Folin-Ciocalteu were examined by comparing to those of
colorimetric method (Singleton and Rossi, known antioxidants such as BHT (butylated
1965), calibrating against gallic acid standards hydroxytoluene), BHA (Butylated hydroxya-
and expressing the results as mg gallic acid nisole) and trolox by 1, 1-diphenyl-2-
equivalents (GAE g-1) extract for seed and skin picrylhydrazyl (DPPH) from Sigma (St. Louis,
extracts. Data presented are average of three MO, USA) using the method of Shimada et al.
measurements. (1992). Briefly, a 1.0 ml solution of the
samples (seed and skin extracts) and standards
HPLC determination of phenolic compounds at 100 µg ml in methanol was mixed with 1.0
Chromatographic analyses were carried out on ml of methanolic solution of DPPH (0.2 mM).
a Shimadzu model HPLC system (Shimadzu The mixture was shaken vigorously and
Corp., Kyoto, Japan). Separation of phenolics allowed to stand at room temperature for 30
was performed by the modified method of min. Then the absorbance was measured at 517
Caponio et al. (1999). Reversed phase (RP)- nm against methanol as the blank in a PG
HPLC analysis was done using a SCL-10Avp Instruments T70 Plus Dual Beam Spectropho-
system controller, a SIL- 10AD vp tometer (Arlington, MA, USA). The addition of
autosampler, a LC-10AD vp pump, a DGU-14a the samples to the DPPH solution caused a
degasser, a CTO-10 A vp column heater, and a rapid decrease in the optical density at 517 nm.
318
The degrees of discoloration indicate the Determination of reducing power
scavenging capacity of the samples. Lower The reducing power of samples were deter-
absorbance of the reaction mixture indicated mined by Oyaizu method (1986). Absorbance
higher free radical scavenging activity. The of supernatant was measured at 700 nm and
effect of antioxidant on DPPH radical compared to three standards, BHA, BHT and
scavenging was thought to be due to their trolox; any increase in absorbance is
hydrogen donating ability or radical scavenging synonymous of an increase in reducing power.
activity (Baumann et al., 1979). Antioxidants
break the free radical chain of oxidation and RESULTS AND DISCUSSIONS
donate hydrogen from the phenolic hydroxyl
groups. Therefore, the resulting stable Total phenolic compound content, antiradical
endproduct does not permit further oxidation of and antioxidant activity of seed and skin are
the lipid (Sherwin, 1978). All determinations presented in Table 1.
were done in triplicate and the percent of As it is observed in Table 1, the yields (dry
DPPH decolouration of the samples were weight) of grape seed and skin had 12.60% and
calculated according to the formula: 9.64 %, respectively.
Antiradical activity (%) = 100x[(absorbance of
control-absorbance of sample)/absorbance.
Table 1. Yield, total phenolic compound content, antiradical and antioxidant activity
of seed and skin of ʻSenirkent Karasiʼ grape cultivar
Sample Yield Total phenolic DPPH Reducing power ( µgl-1)
(%) content (mg/g GAE) (100 ppm extract (Absorbance)
(%)
250 ppm 1000 ppm
Seed 12.60±0.63 52.32±3.25 95.90±1.03 1.64±0.18 2.39±0.20
Skin 9.64±0.68 1.89±0.29 16.22±0.80 0.08±0.00 0.31±0.01
Total phenolic contents of the samples were and skin extracts were determined by HPLC
estimated with Folin-Ciocalteu colorimetric and presented in Table 2.
method. When total phenolic contents of seeds
extracts were calculated as mg GAE g-1 (Table Table 2. Phenolic compounds of seeds and skin of
1) it is found that seeds had higher total ʻSenirkent Karasiʼ grape cultivar
phenolic compound content than skins. Seeds Phenolic compound Skin, µg. g-1 Seeds, µg. g-1
and skin had total phenolic compound content Gallic acid 5.29±0.10 144.76±0.45
of 52.32±3.25 and 1.89±0.29 mg/g, respec- (+)-Catechin 3.43±0.21 637.88±5.55
(-)-Epicatechin nd 746.94±2.13
tively, in terms of gallic acid. Results are in Caffeic acid 3.83±0.29 nd
agreement with those found by (Negro et al., Syringic acid 17.01±0.21 nd
2003; Yılmaz and Toledo, 2004; Iacopini et al., p-coumaric acid 0.70±0.01 nd
2008). These researcher also found that seeds Trans-Resveratrol 2.47±0.02 nd
had higher total phenolic compound content Ouarcetin 2.49±0.20 nd
Kaemferol 0.50±0.02 nd
than skin.
HPLC method for analyzing phenolics in the
samples has some advantages, such as easy and It is found that skin had higher number of
time consuming procedure for preparation of phenolic compounds than seeds. In skin
the samples, possibilities of quantification of a samples 8 compounds such as gallic acid,
greater amount of diverse phenolics, the catechin, caffeic acid, syringic acid, p-coumaric
precision, accuracy and detection limits acid, resveratrol, quarcetin and kaempferol
obtained for the phenolics quantified by this were detected whereas in seed samples only 3
method enabling its application to grape compounds such as gallic acid, epicatechin and
(Gomez Alonso et al., 2007). The amounts and catechin were detected. Gallic acid amount in
variations of phenolic compounds in the seed seeds and skin were determined as 144.76±0.45
µg g-1 and 5.29 ±0.10 µg g-1, respectively. In
319
the same manner catechin amount in seeds and µg l-1 and 2.39 µg l-1values at 250 and 1000 ppm,
skin were determined as 637.88 ±5.55 µg g-1 respectively, whereas skin had 0.08 µg l-1 and
and 3.43 ±0.21 µg g-1, respectively. As regards 0.31 µg l-1 reducing power ability values at 250
to the presence of catechin in skin and seeds, it and 1000 ppm, respectively. Higher absorbance
is commonly known that flavan-3-ols are loca- values correspond to higher reducing power
ted in both grape skin and seeds; however, skin thus it is found that seeds had higher reducing
contains much lower concentrations of flavan- power than skin. Hua et al. (2008) reported that
3-ols than seeds (Revilla and Ryan, 2000). in seeds of grapes there was a correlation
In addition, another flavonoid, epicatechin, between reducing power. In this research it is
amounted in seeds 746.94±2.13 µg g-1 and it also found that seeds had higher total phenolic
was not detected in skin. The results agree with compound content than skins and seeds had
the studies of Cheynier (1998), Rodriquez higher reducing power than skin.
Montealegre et al. (2006) and Baydar et al.
(2011), who also found that grape seeds had CONCLUSIONS
higher flavanol contents than skins. Another
study also found that there was presence of In this research we determined phenolic
epicatechin in seeds, whereas there was no compounds, antiradical and antioxidant activity
epicatechin in skin (Souquet et al., 2000). in seeds and skin of ʻSenirkent Karasiʼ grape
Trans-Resveratrol, a phytoalexin that belongs cultivar which is commonly produced and
to the group of compounds known as stilbenes, consumed in Isparta province.
is known to occur in grapes and consequently The results obtained in this study showed that
in grape products and in wine. Trans- large differences were found grape seed and
resveratrol was found in 2.47 µg g-1 in the skin skin in relation to the phenolics composition.
extracts. Baydar et al. (2011) also found 1.82 Senirkent Karası grape’s seeds, and skins con-
and 4.02 mg 100 g-1 of trans-resveratrol in tained different phenolics with different levels
grape skin extract. Iacopini et al. (2008) and these variations affected the antioxidant
explained this result as the consequence of the capacity of the samples. Total phenolic con-
fact that grapes produce stilbenes in response to tents, reducing powers of grape seed extracts
mold infections and physiological stresses. If are higher than those of grape skin extracts.
these stresses are not present, the levels of The result of study is important because grape
stilbenes in grapes remain low. seeds and skin are a good source of phenolic
Radical scavenging activities of grape extracts, compounds that have positive effect on health,
and standards were tested by the DPPH and they are rich in natural antioxidants. Thus,
method. When radical scavenging activities of determining these compounds in a local
seed and skin is examined, it is observed that cultivar is important in terms of health issue.
seeds had 95.90% antiradical activity whereas
skin had 16.22% antiradical activity. The ACKNOWLEDGEMENTS
radical scavenging activities of the seed
extracts were considerably higher than those of This research work was carried out with the
skin extracts. Grape seed extracts almost support of Suleyman Demirel University,
completely inhibited DPPH absorbtion. Coordination Center for Scientific Research
Otherwise skin extract contained remarkably Project, and also was financed from Project
lower amounts of radical scavenging 2551-M-10
compounds. Some researcher reported that
there was a correlation between DPPH activity REFERENCES
and total phenolic compound content of seed
(Guendez et al., 2005; Hua et al., 2008). In this Ayed N., Yu H., Lacroix M., 1999. Improvement of
research it is also found that seeds had higher anthocyanin yield and shelf-life extension of grape
pomace by gamma irradiation. Food Research
total phenolic compound content than skins and International, 539-43.
seeds had higher DPPH activity than skin. Baumann J, Wurm, G., Bruchhausen, F., 1979.
When the reducing powers of seeds and skin Prostaglandin synthetase inhibiting O2-radical
was examined it was found that seeds had 1.64 scavenging properties of some flavonoids and related
320
phenolic compounds. Naunyn-Schmiedebergs Negro C., Tommasi A., Miceli A., 2003. Phenolic
Archives of Pharmacology, 30: 27-32. compounds and antioxidant activity from red grape
Caponio F, Alloggio V., Gomes T., 1999. Phenolic marc extracts. Bioresour. Technology, 87, 41-44.
compounds of virgin olive oil:Influence of paste Oyaizu M., 1986. Studies on products of browning
preparation techniques. Food Chemistry 64: 203-209. reaction: antioxidative activities of products of
Cheynier V., Labarbe B., Moutounet M., 2001. browning reaction prepared from glucosamine.
Estimation of proantho proanthocyanidin chain Japanese Journal of Nutrition, 44, 307–315.
length. Methods in Enzymology, 335, 82-87. Revilla E., Ryan J., 2000. Analysis of several phenolic
Claussen T.P., Reichardt P.B., Bryant J.P., Provenza F., compounds with potential antioxidant properties in
1992. Condensed tannins in plant defense, a grape extracts and wines by HPLCphotodiode array
perspective on classical theorses, Plant Polyhends, detection without sample preparation. Journal of
Ed. By R.W. Mennengway and P.E. Laks Plenum, Chromatography A 881: 461- 469.
New York. Rodriguez Montealegre R., Romero Peces R., Chacon
Gomez-Alonso S., Garcia R.E., Hermosin Gutierrez I., Vozmediano J.L., Martinez Gascuena J., Garcia
2007. HPLC analysis of diverse grape and wine Romero E., 2006. Phenolic compounds in skins and
phenolics using direct injection and multidetection by seeds of ten grape Vitis vinifera varieties grown in a
DAD and fluorescence. Journal of Food Composition warm climate. Journal of Food Composition and
and Analysis, 20: 618-626. Analysis. 19, 687-693.
Guendez R., Kallithraka S., Makris D.P., Kefalas P., Shimada K., Fujikawa K., Yahara K., Nakamura T.,
2005. Determination of low molecular weight 1992. Antioxidative properties of xanthan on the
polyphenolic constituents in grape (Vitis vinifera sp.) autoxidation of soybean oil in cyclodextrin emulsion.
seeds extracts: Correlation with antiradical activity. Journal of Agricultural and Food Chemistry, 40, 945-
Food Chemistry, 89: 1-9. 948.
Hua L., Xıaoyu W., Peıhong L., Yong L., Hua W., 2008. Singleton V.L., Rossi J.R., 1965. Colorimetry of total
Comparative study of antioxidant activity of grape phenolics with phosphomolibdic - phosphothungstic
(Vitis vinifera) seed powder assessed by different acid. American Journal of Enology and Viticulture,
methods. Journal of Food and Drug Analysis, 16 (6): 16, 144-158.
67-73. Souquet J.M., Labarbe B., Le Guerneve C., Cheyneir V.,
Iacopini P, Baldi M., Storchi P., Sebastiani L., 2008. Moutounet M., 2000. Phenolics composition of grape
Catechin, epicatechin, quercetin, rutin and resveratrol stems. Journal of Agricultural and Food Chemistry,
in red grape: Content, in vitro antioxidant activity and 48, 1076-1080.
interactions. Journal of Food Composition and Yilmaz Y., Toledo R.T., 2004. Major flavonoids in grape
Analysis 21: 589-598. seeds and skins: Antioxidant capacity of catechin,
Jayaparakasha G.K., Selvi T., Sakariah K.K., 2003. epicatechin, and gallic acid. Journal of Agricultural
Antibacterial and antioxidant activities of Grape and Food Chemistry, 52: 255-260.
(Vitis vinifera) Seed Extracts. Food Research Yi W., Akoh C., Fischer J., Krewer G., 2006. Effects of
International, 36, 117-122. phenolic compounds in blueberries and muscadine
Macheix J.J., Fleuriet A., Billot J., 1990. Fruit Phenolics, grapes on hepg2 cell viability and apoptosis. Food
CRC Press. Boca Raton, FL. Research International, 628-638.
321
322
STUDY ON THE BEHAVIOR OF THE VICTORIA TABLE GRAPE

VARIETY IN THE HINOVA VITICULTURAL AREA,
MEHEDINTI COUNTY
Liviu Teodor MUNTEAN
University of Craiova, Faculty of Horticulture

A. I. Cuza Street, No. 13, Craiova, Dolj County, Romania
Abstract
This paper aims at studying the favorability of the cultivation of the ʻVictoriaʼ table grape variety in the Hinova
viticultural area, Mehedinti County, in a non-irrigated system. The research was carried out in 2017. After determining
the viability of the buds losses were found, so there were studied 3 different fruit loads to follow the number of buds
started from the vegetation and to meet the number of shoots required for the study. Once this requirement has been
fulfilled, the work of normalizing the inflorescences was carried out, taking into consideration 5 variants, namely: 6, 8,
10, 12, 14 inflorescences/vine. The evolution of these fruit variants was followed up until reaching the full maturity
stage, with the harvesting of the grapes resulting in the deliverable production of each variant of the studied fruit load.
The conclusion of this study is that Hinova viticultural area in Mehedinti County is favorable to the cultivation of the
ʻVictoriaʼ vine variety in a non-irrigated system, only if a moderate fruit load is respected, thus obtaining quality
grapes.
Key words: favorability, Hinova, table grapes, ʻVictoriaʼ variety, viticultural area
Hinova viticulture area located in Mehedinti In order to study the favorability of the
County enjoys a climate with Mediterranean cultivation of the Victoria table grape variety in
influence, due to its proximity to the Danube, the Hinova viticultual area located in Mehedinti
mild winters and dry summers (Gherasim et al., County, the following elements were studied:
1977). the viability of the winter buds in order to
This very important factor in the zoning of vine determine the type of winter pruning that had to
varieties allows the cultivation of table grape be carried out, the number of winter buds to be
varieties in this area, who has been cultivated assigned to the fruit elements (2 canes + 2
until 1989 with table grape varieties such as spurs), the number of buds started in
ʻPerla de Csabaʼ, ʻRegina Viilorʼ, ʻCoarna vegetation, the number of inflorescences
albaʼ, ʻCoarna neagraʼ, ʻChasselas doréʼ, assigned to each vine, the determination of the
ʻChasselas rozéʼ. The table grape variety weight of the harvested grapes.
Victoria has special charactheristics (precocity, This study was conducted during the year 2017.
size, weight, taste, appearance), being grown
on large surfaces both in Romania and in RESULTS AND DISCUSSIONS
countries with a tradition in the cultivation of
table grape varieties: Italy, Greece, Spain, Knowing the viability of the winter buds is an
Turkey (Giugea et al., 2015) important element in determining the number
The study of the behavior of the ʻVictoriaʼ table of winter buds to be attributed to the fruit
grape variety in the Hinova viticultural area in a elements, with the occasion of the winer
non-irrigated system is a novelty for this area, pruning. It is done by cross-section with the
since it has not been cultivated in the past, so it blade by the buds during the rest period of the
can be noticed what technological measures vines, once the danger of other losses due to
must be taken to obtain the best productions, low temperatures has passed. Once the viability
both quantitatively and qualitatively. check has been carried out, it has resulted a
323
percentage of loss of the main buds due to Table 1. Load variants/vine
winter frost of 60%. No. No. No. buds No.
variant winter started in inflorescences/vine
buds vegetation
1 16 13 20
2 20 12 19
3 24 14 23
Analyzing the data presented in this table, we

can conclude that from of all the variants taken
in the study resulted a number of similar
shoots, the same conclusion can be made in the
case of the number of inflorescences distributed
on each grape vine. Next, there are 5 variants of
fruit loads taken in study, obtained by the work
of normalization of the inflorescences: 6, 8, 10,
12, 14 inflorescences/grape vine.
Figure 1. Cross section made with the blade through the
bud complex
The type of winter pruning is double Guyot

(2 woody shoots + 2 spurs) .
Figure 3. Buds widening phenophase
The evolution of the 5 variants was followed during

the vegetation period, of interest being the period of
ripeness and the period of maturity of the grapes
Figure 2. Guyot type pruning applied (Table 2).
to the ʻVictoriaʼ vine
In order to study the number of buds that will

start in vegetation and to meet the number of
shoots with fruit necessary for the study, we
have studied 3 different variants of loads,
which will be attributed to the fruit elements
during the winter pruning, and namely: 16
winter buds/grape vine divided into 2 canes x 6
winter buds + 2 spurs , 20 winter eyes/grape
vines divided by 2 canes x 8 winter buds + 2
spurs, 24 winter buds/grape vines divided by 2
canes x 10 winter eyes + 2 spurs.
The number of buds started in vegetation as
well as the number of inflorescences / grape
vine for each studied variant are presented in Figure 4. Phenophase of 2-3 leaves
Table 1:
324
Table 3. Number of harvested grapes and weight/vine
dending on the number of infloreces left on the vine
No.inflorescences/vine No. of Total grape
harvested weight/ vine
grapes
6 6 4.2
8 8 5.4
10 9 4.9
12 9 4.6
14 4 1.4
Figure 5. Blooming phenophase
Figure 7. ʻVictoriaʼ vine with a load of 12 grapes/vine
Figure 6. Ripness phenophase Figure 8. ʻVictoriaʼ vine with a load of 8 grapes/vine

Table 2. Ripeness/maturity date of grapes depending on
the number of inflorences left on the vine CONCLUSIONS
No.inflorences/vine Ripeness starting Date of
date maturity This study highlights the fact that once the
6 July 23-24 August 10 number of inflorescences / grapes on the grape
8 July 22-24 August 14 vine increases, it increases also the date when
10 July 26-27 August 16
the grapes reach maturity, and the number of
12 July 29-30 August 22
14 August 1-2 August 25 grapes harvested from the grape vine decreases.
It was noteworthy that the best results had the
In Table 3 there are presented data showing the grape vines with a load of 8-10 grapes/vine.
number of grapes harvested from the three variants Also, it was observed that in the variants with
taken into study as well as the total weight of fewer grapes/vine they were bigger and more
grapes obtained/grape vine: compact, and in the variants with more
grapes/vine they were smaller and less
compact.
325
The conclusion of this study is that the Hinova REFERENCES
viticultural area located in Mehedinti County is
favorable to the cultivation of the ʻVictoriaʼ Gherasim C., Aurel G., Gheorghiu A., 1977. Drumurile
vine variety in a non-irrigated system, only if a viei şi vinului în România. Editura Sport-Turism,
Bucureşti.
moderate fruit load is respected, thus obtaining Martin T., 1978. Cultura neprotejată a viţei-de-vie.
quality grapes. Editura Ceres, Bucureşti.
Oslobeanu M., Oprean M., Alexandrescu I., Georgescu
ACKNOWLEDGEMENTS M., Banita P., Jianu L., 1980. Viticultura generală şi
specială. Editura Didactică şi Pedagogică, Bucureşti.
Popa A., Giugea N., Genoiu C.T., 2015. Oltenia, mica
This research work was carried out with the Românie viticolă. Editura Aius, Craiova.
support of the Muntean table grape vineyard Kunin D., 1979. Fiziologia vinului. Editura Ceres,
located in Hinova, Mehedinti County, Romania Bucureşti.
326
CONSIDERATIONS REGARDING THE USE OF ANTI-HAIL NETS

FOR THE PROTECTION OF TABLE GRAPE PLANTATIONS
IN HINOVA VITICULTURAL AREA, MEHEDINTI COUNTY
Liviu Teodor MUNTEAN
University of Craiova, Faculty of Horticulture

A. I. Cuza Street, No. 13, Craiova, Dolj County, Romania
Abstract
This paper aims to present the benefits of implementing a protection system with nets against hail in table grape
vineyards, as well as the problems that may occur. The study was carried out between 2013 and 2017, in a table grape
vineyard. in Hinova, Mehedinti County. It was possible to analyze during the 5 years of study the behavior of vegetative
phenophases of the ʻVictoriaʼ vineyard grown in a system covered with the anti-hail net, compared to another
ʻVictoriaʼ vineyard cultivated in an uncovered system. In the studied period, hailstones occurred in 2014, 2016 and
2017, the most severe hail fall occurring on June 20, 2014, where the damage to the unprotected ʻVictoriaʼ vineyard
was 100%. In conclusion, the protection of the vineyards with anti-hail net is beneficial, it protects to a certain extent
against late spring frost, but it also draws certain shortcomings or reservations in deciding to choose for a such
protection: high-cost investment, more frequent application of treatments to fight diseases and pests.
Keywords: anti-hail net, Hinova area, Mehedinti County, vine protection.
INTRODUCTION
Hailstorms, according to their intensity, can

have destructive effects on vine plantations,
causing significant damage, compromising the
grape production in that year, plant tissue
damage, thus creating doorways for the
diseases of the vine and negatively influencing
the resistance to winter frosts (Bernaz, 2003).
Faced with these unpredictable natural
phenomena, grape growers in general and table
grape growers in particular can take a series of
measures, one of which is the installation of
nets to protect against hail. Taking this into Figure 1. ʻVictoriaʼ vineyard protected by anti-hail net
account, the present work presents the benefits
that such protection can offer, as well as the
problems that can cause in a table grape
vineyard.
In order to highlight the advantages and

disadvantages offered to vineyards by anti-hail
net systems, two plots cultivated with
ʻVictoriaʼ table grapes variety were studied,
one having a net protection system against the
hail installed, the other not having this system
installed. Figure 2. Unprotected ʻVictoriaʼ vineyard
327
The analysis period in this study was compromised, destroying both grape bunches,
2013-2017. leaves and vine shoots, who suffered deep
tissue lesions.
The hail protection system was installed in the

summer of 2013. During the year 2013 no
hailstones occurred.
The two plots behaved relatively similarly until
ripeness (BBCH 81-85), when in the plot
covered with the net against the hail there were
noticed botrytis infested grapes, especially in
the middle part of the plot, while on the
uncovered vineyard the grapes were perfectly
healthy. The net against hail was removed at
the end of October.
In 2013, both plots were treated with identical
phytosanitary treatments.
In 2014, the anti-hail net was installed in early
April. Both plots were treated with identical
Figure 4. Damage caused by heavy hail fall
phytosanitary treatments until the beginning of
blooming. During the beginning of the In the protected vineyard, with the exception of
blooming (BBCH 55-57), in the plot covered the marginal rows, there were no problems
with the net there were reported inflorescences caused by hail, thus protecting both the grape
attacked by mildew, although the number of production and the health of the vines.
treatments and the interval between them was
strictly respected, using only phytosanitary
products with systemic action. Measures have
been taken and additional treatments were
administrated to keep the attack under control.
There was no mildew attack reported in the
uncovered plot.
Figure 5. Hail stones on top of the anti-hail net system
Figure 3. ʻVictoriaʼ inflorescence attacked by mildew
On June 20, during a strong storm accompanied

by massive hailstones, grape production in the
uncovered ʻVictoriaʼ vineyard was totally Figure 6. Average-size hailstones reported
after hail fall in 2014
328
In 2015, on the vine shoots from the ʻVictoriaʼ the production of grapes has been low and of
vineyard covered with anti-hail net traces of poor quality.
excoriosis attack was reported, vine wood
disease never met before in the plantation. The
uncovered plot did not show signs of this
disease.
During the year, high-pressure infection of
mildew, oidium and botrytis was reported in
the anti-hail protected vineyard. In order to
maintain the state of health of the vineyards,
the interval between phytosanitary treatments
was shortened, and at the same time in the
disease control program there were introduced
phytosanitary products with contact action for
additional protection in critical moments
(copper oxychloride against mildew and
soluble sulfur against oidium). This increased Figure 7. Temperatures of 0oC recorded
by min-max thermometer on April 22, 2017
the number of phytosanitary treatments applied
to the vine throughout the wine year.
Hailstones did not occur in 2015.
The year 2016 was similar to 2015, on the plot
covered with anti-hail net, the total number of
treatments applied against diseases and pests
increased to 10, compared to 7 treatments
applied on the plot not covered with anti-hail
net.
Two hailstorms occurred at the beginning of
May and early July, of low intensity, resulting
in minor damage to the unprotected plot.
August was rainy, with cumulative rainfall
reaching 70 L/sqm. These climatic conditions,
which intersected with the time when the Figure 8. ʻVictoriaʼ young shoots affected
grapes reached maturity, gave rise to a strong by late-spring frost cultivated in unprotected vineyard
botrytis attack, with considerable losses of
production being reported in the covered plot
with 60% compared to 20% - losses in the
uncovered plot.
In the spring of 2017, on April 22, while the
vine shoot was in the 2-3 leaf stage (BBCH10-
12), the min-max thermometer recorded a
minimum of 00C during the night, resulting in
producing a late spring frost. As a result of this
climatic accident, the vine shoots of the
uncovered plot were affected in a proportion of
75-80%.
On the plot covered with the anti-hail net there
was no significant damage, the production of Figure 9. ʻVictoriaʼ young shoots unaffected
grapes being a normal one, characteristic of the by late-spring frost cultivated in net-protected vineyard
variety. The shoots on the plot uncovered
against hail have been partially recovered, The number of phytosanitary treatments
starting from the buds situated at the wrist of applied to the protected plot was 10, and in the
the leaves , but they were weak, had no fruit, so unprotected plot 6.
329
the late spring frosts, when the temperatures are
not well below the freezing threshold.
Among the less pleasant aspects of this study
can be summarized: large initial investment,
reduced ventilation at the level of the hub in the
area covered with anti-hail net, high moisture
content, as well as persistent dew for a longer
time, compared to the plot uncovered with anti-
hail net, which results in the need for a higher
number of phytosanitary treatments.
ACKNOWLEDGEMENTS
Figure 10. Phytosanitary treatment applied in ʻVictoriaʼ This research work was carried out with the
vineyard protected by anti-hail nets support of the Muntean table grape vineyard
located in Hinova, Mehedinti County,
During the year 2017, several hailstorms of Romania.
small and medium intensity occurred, affecting
to some extent the quality of the poor grape REFERENCES
production in the plot unprotected with anti-
hail nets. Bernaz G., 2003. Refacerea viilor vătămate de accidente
climatice şi boli.. Editura M.A.S.T., Bucureşti..
CONCLUSIONS Martin T., 1978. Cultura neprotejată a viţei-de-vie.
Editura Ceres, Bucureşti.
Moser L., 1980. Viticultura modernă. Editura Ceres,
Protecting the vineyards with the anti-hail net is Bucureşti.
an ideal solution for table grape growers who Oslobeanu M., Oprean M., Alexandrescu I., Georgescu
want to obtain a high quality grape production, M., Banita P., Jianu L., 1980. Viticultura generală şi
thus providing a system that can protect both specială. Editura Didactică şi Pedagogică, Bucureşti.
Popa A., Giugea N., Genoiu C.T., 2015. Oltenia, mica
grape production and vegetation year by year Românie viticolă. Editura Aius, Craiova.
by the devastating effects of hailstones, good Kunin D., 1979. Fiziologia vinului. Editura Ceres
results also being obtained in the protection of Bucureşti.
330
EVALUATING SOME OLFACTORY CHARACTERISTICS OF WINE IN

THE CONTEXT OF GLOBAL WARMING IN THE “PLAIURILE
DRÂNCEI” VITICULTURAL REGION
1 1 1
Alina-Mihaela NISTOR , Liviu-Gabriel GRIGORICĂ , Cristina-Elena SCUTĂRAȘU ,
Ștefan-Dragoș COTAN1, Valeriu V. COTEA1, 2, Marius NICULAUA2
1
“Ion Ionescu de la Brad” University of Agricultural Sciences and Veterinary Medicine Iasi,
3 Mihail Sadoveanu Alley, 700490, Iași, Romania
2
Research Centre for Oenology, Romanian Academy Iasi Branch,
9 Mihail Sadoveanu Alley, 700490, Iași, Romania
Abstract
Global warming is recognized among the scientific community as a worrying fact for the environment, which generally
speaking causes unwanted changes in the species’ evolutionary accommodation mechanism to superior thermal
conditions. In this context, this study’s purpose is to point out some chemical transformations which positively
contribute to the olfactory characteristics of wine. Global warming determines an increase in the alcohol levels,
therefore causing a proportional increase of secondary fermentation products, which are helpful in giving a pleasant
aroma. The samples used for the analysis were of the ʻSauvignon Blancʼ variety from the "Plaiurile Drâncei" vineyard.
The alcohol concentration of the samples varies between 11.8 and 13.6% v/v. The identification of chemical compounds
was performed using the head space technique coupled with GC-MS. Following the analysis, there were identified
chemical compounds which mainly give floral, fruity, citric notes and also not specific to fermentation, moreover giving
a pleasant olfactory sensation to the final product. Therefore, the olfactory characteristics of wine depend on a series of
factors beginning from climate parameters and ending with the fermentation process.
Key words: wine, GS-MS, global warming
INTRODUCTION “Oravița”, “Rogova”, “Drâncea” and

“Oprișor”. Around the year 1900, during the
Romania is country rich in soil and diverse visit of King Charles I, the wines of this
microclimates. It is for these reasons why from vineyard became renowned in the whole
the country’s territory there can be obtained country, being sought after by every “collector”
from white wines characterized by freshness of good wines. These wines are characterized
and lean fruity aromas to red wines, noble, by high levels of alcoholic concentration and
suited for aging and also wines with low or rich in mineral substances and extract.
high acidity and even wines suitable for The vineyard is situated on slopes, divided into
producing sparkling wines. Therefore, unlike in two regions: the North West region of the fields
other countries, the climate of Romania allows of “Oltenia” which descends towards the
for variation in the types of vinification. Danube in several stages which have altitudes
(Stoian, 2011; Cotea et al., 2003). smaller than 200m; and the region around the
The “Plaiurile Drancei” vineyard is located in “Bălăcița” plateau which has altitudes between
the hilly terrain of the fields of “Oltenia”, 200m and 300m. The tilt of the slopes varies
towards the Danube and is part of the between 5 to 10 degrees. This placement is one
vinicultural region of the “Getic” plateau and is of the factors that contribute to the superior
one of the most southern vineyards of quality of the wines. Another factor is the soil’s
Romania. Documentary certified sources composition. Their composition is made of:
indicate the fact that viticulture began to 60% brown-red strongly eroded soil, 15%
develop in this area beginning around 1407, the ordinary brown-red soil, 10% brown-red
most well-known vinicultural locations being
331
eroded soil, 10% ordinary leached chernozem when coming to a higher degree of ripening
and 5% ordinary alluvial soil. they can give floral and exotic fruity tones.
Ion Ionescu de la Brad claimed in one of his Therefore the range of aromas can vary from
papers that in “Oprișor” the sun falls on the forest apples, grapefruit, passion fruit, kiwi and
vineyard all day long and that the soil in this well ripened cantaloupe. Furthermore, the
locality was “red clay”, referring to the brow- specifics of this variety consist of the smell of
red soil. The hydrography is characterized by a “cat urine”, but experienced in a pleasant way
network of small rivers and their tributaries. which is a designator of quality and is used as a
In these vinicultural areas the clime is means of identification. Although it is a variety
continental temperate with mediterranean that offers herbal (green leaves) tones, this
influences and is characterized by hot summers changes when the level of alcoholic
and relatively mild and wet winters (the frost concentration rises or oak is used in the
seldom happens or it is of weaker intensity). winemaking process and the exotic tones
The average temperature is between -1 and -2 prevail (Old, 2013; Stoian, 2011; Cotea et al.,
degrees Celsius in January and between 21 and 2003).
22 degrees Celsius in July. The average annual In the context of global warming this paper
temperature is 11 degrees Celsius and the takes a more positive approach on the changes
average annual precipitations are between 550 caused by the climate factors. The trend is that
and 600 mm, while snow is relatively scarce. of the vineyard, through its characteristics, to
During the periods of drought there is a water be able to adapt to the sharp rise of
deficit. By taking into account all these temperatures, respectively resisting to the
parameters of the natural setting, they add up to absence of precipitations. If a variety of grapes
an ecosystem that is favourable for high quality is developing naturally, while keeping its
viticulture (Cotea and Andreescu, 2008; Cotea specifics, in a certain range of temperature,
et al., 2003; Toti et al., 2017; Ionescu, 1968). when the temperature rises on average even
According to the National Meteorology with as little as 1-2 degrees Celsius there is a
Administration’s estimations, which were risk of losing its defining attributes or not being
presented in a report, the average annual able to adapt at all in these new climate
temperature of Romania increased during 1961- conditions. In that case the approach taken by
2014 by 0.5 degrees Celsius and regarding the winemaker is very important. The biggest
precipitations, their quantity resulted in an change in wines that is caused by global
increase in the number of extremely droughty warming is the increase in the alcoholic
years. The forecasts on this subject show that in concentration (even with as much as 2-3%).
the following years (2021 - 2050) the summers Once this happens, the intensity of the aroma
will become hotter, draughty and the also increases, especially the aromas of oven
precipitations will decrease by up to 10-20% on baked fruits, exotic aromas, spicy or oak
average (Sandu, 2015). aromas. The current trend of the Romanian
The ʻSauvignon Blancʼ variety is the second consumer is to drink white young wines which
most popular white wine in the world and is are as floral, fruity and fresh as possible. The
native to France, Bordeaux but it is grown from formation of these aromas can be better studied
the oldest times in the Loire Valley, in New through volatile chemical compounds, which
Zeeland or in the USA. It belongs to the are responsible of the product’s final result
category of quality wines. Due to its (Jones, 2007; van Leeuwen, 2016).
adaptability, in present it is being grown in a lot
of countries and on all the continents. It is MATERIALS AND METHODS
assumed that the name “Sauvignon” comes
from the French word “sauvage” which means The samples belong to the ʻSauvignon Blancʼ
untamed or wild, the association being made variety from the vinicultural region of the
with the wild vines or with its special aroma. “Getic” plateau, “Plaiurile Drâncei” vineyard,
Generally, it is a wine with a soft aroma. Based Oprișor locality, and were obtained in three
on the grapes’ degree of ripening they can give different years: 2013, 2014 and 2015. A
herbal, vegetable or green fruity tones, and number of five samples was chosen for this
332
study (denoted SB0-SB4) and the difference connected with the increase of alcoholic
between each other consists of the type of yeast concentration. 2014 was an unusual year, being
strain used for the fermentation process. considered by the National Meteorology
Sample SB0 was obtained through gravitational Administration as the fourth rainiest year from
clarification in the absence of yeast and the rest the last fifty years. This caused a decrease in
of the samples (SB1, SB2, SB3, SB4) were temperatures and July did not bring any heat
obtained through gravitational enzymatic waves as it was expected (Polifronie, 2014).
clarification. Therefore, following a comparison between the
The samples from 2015 were chosen for a more samples’ alcoholic concentration, it was
thorough analysis, from which a volume of 7 expected that the wine from 2015 would be
mL was collected and analysed through gas richer in volatile compounds which give more
chromatography using the head-space intense fruity aromas. This was confirmed by
technique coupled to a mass spectrometer. The the results from the chromatographic analysis
equipment used was a Shimadzu Head Space (Table 2). There were several volatile
HS 20 trap - GC 2010plus-MS8040TQ. The compounds that were identified by this
compound identification was done by analysis, but are showed only those for which
correlating with the software’s database and the the literature gives clear information of their
quantitative results were obtained using the flavour (Wu, 2016; *thegoodscentscompany)
internal standard method by comparing with 4- and especially those which have a positive
methyl-2-pentanol (Grigorică, 2017). The impact and through which the influence of
samples’ alcoholic concentration was alcoholic concentration can be observed.
determined according to the OIV-MA-AS312- By analysing the aromatic profile it is obvious
01 method (OIV, 2018). that the dominating aromas are fruity.
Responsible for this are the chemical
RESULTS AND DISCUSSIONS compounds from different classes such as
alcohols, esters and carboxylic acids. Their
After determining the alcoholic concentration concentration in this case is also influenced by
of all the samples, the values from Table 1 the yeast that was used for fermentation - this
were obtained: explains the difference between the values of
the same compound and with the same
Table 1. Alcoholic concentration of samples for year of alcoholic concentration (for example in the
production 2013-2015
case of ethyl octanoate for samples SB2 and
Year of Alcohol SB3 or in the case of hexanoic acid for the
Samples
production (%v/v)
same samples).
2013 SB0 12.8
2013 SB1 13.1
Most of the chemical compounds may have
2013 SB2 13.1 several sensorial characteristics (from pleasant
2013 SB3 13.0 fruity aroma to fatty and waxy repelling
2013 SB4 13.0 aromas), depending on the concentration in
2014 SB0 11.8 which they are present (Wu, 2016). From the
2014 SB1 12.0 numerical data obtained it follows that ethyl
2014 SB2 12.1 octanoate and ethyl decanoate have the biggest
2014 SB3 11.9
2014 SB4 12.0
influence in forming the fruity aromas. The
2015 SB0 13.1 specifics of these aromas are apricot, banana,
2015 SB1 13.6 pear, pineapple, apple or wine.
2015 SB2 13.3 Other compounds that present sweet and fruity
2015 SB3 13.3 aromas are 3-methyl-1-pentanol, 2-undecanol,
2015 SB4 13.5 ethyl decanoate, isoamylacetate (banana or
pear), 3-methylbutyl decanoate (banana),
In 2015 there is an increase in alcoholic decanoic acid (citrus), ethyl nonanoate (apple
concentration of approximately 0.5% v/v across or banana), 3-hexenol acetate (banana or
all samples, compared to 2013. This is candy) or ethyl pentadecanoate (honey).
somewhat expected given the climatic changes,
especially the air temperature, which is strongly
333
Table 2 Results of chromatographic analysis for wine samples from 2015
Alcohol concentration (%v/v) 13.1 13.6 13.3 13.3 13.5

Samples SB0 SB1 SB2 SB3 SB4
Chemical compounds Flavour Samples concentration (microequivalents internal standard/L)
isoamylacetate banana, pear ND 260.77 63.34 43.61 ND
2-methyl-1-propanol ethereal, apple, bitter, cocoa, winey ND 22.14 27.18 14.33 ND
2,6-dimethyl-4-heptanone sweet, mint 37.19 52.54 ND ND 44,4
3-hexenol acetate banana, candy, floral, green 36.19 31.97 40.31 30.09 19.56
5-methyl-2-hexanol grassy 7.36 7.63 6.2 6.48 17.9
3-methyl-1-pentanol fruity 2.9 9.89 ND ND 9.6
ethyl octanoate sweet, fruity, apricot, banana, pear, pineapple, wine, waxy, dairy 1309.57 2266.25 1911.94 1057.88 2376.98
1-hexanol floral, green, resin 4.92 24.9 17.93 20.24 14.35
3-hexenol moss, fresh 13.89 93.62 59.48 47.4 ND
2-hydroxyethyl propanoate butter 76.14 59.07 23.42 28.69 156.46
1-hydroxy-2-propanone butter, grassy, malt, spicy 4.88 ND 4.11 ND ND
ethyl nonanoate fruity, apple, banana, rose, winey, cognac, waxy 5.99 20.08 13.49 9.4 17.4
2-undecanol fruity 9.71 27.91 19.72 18.96 24.74
ethyl decanoate sweet, fruity, apple, waxy 2141.92 1848.29 1828.46 1054.08 1211.38
isobutanoic acid acidic, sour, cheesy, dairy, buttery, rancid 50.59 66.34 6.19 6.15 32.2
334
2,3-butanediol fruity, buttery 35.53 29.41 ND ND 11.11
butanoic acid sharp, acidic, dairy, cheesy, sour, buttery with a fruity nuance 17.15 29.62 ND ND 3.56
fruity, passion fruit, apple; apricot; cranberry, peach, pear,
diethyl succinate 7 37.88 36.93 35.8 33.18
chocolate, grape, floral, musty, waxy, earthy
isovaleric acid fruity, cheese, dairy, acidic, sour, pungent 18.27 36.97 28.98 25.3 30.18
ethyl palmitate fruity, waxy, creamy and milky with a balsamic nuance 295.06 520.17 402.47 386.69 339.95
3-methylbutyl decanoate sweet, fruity, banana, waxy, green nuance 5.91 10.49 9.31 9.42 4.14
hexanoic acid sour, fatty, sweat, cheese 271.96 345.82 312.73 274.28 268.49
2-phenylethanol sweet, floral, fresh and bready with a rose, honey nuance 225.19 237.77 251.38 251.17 255.68
ethyl myristate sweet, violet, iris, waxy ND 5.04 ND 31.48 ND
octanoic acid waxy, dirty, cheese, phenolic, fatty, oily 728.65 864.24 784.73 792.78 703.07
ethyl pentadecanoate sweet, honey ND 12.03 ND ND 6.21
decanoic acid citrus, unpleasant, rancid, sour, fatty 312.55 162.56 141.03 114.26 99.43
ethyl tetradecanoate mild, waxy ND 10.13 13.71 11.12 8.89
ethyl linolate fruity, mild, fatty ND 8.3 5.35 5.83 7.2
*ND - not detected
Some compounds may have a specific wine REFERENCES
aroma or in the case when the samples evolve
in time (in gets older in a good way) there may Cotea D.V., Barbu N., Grigorescu C., Cotea V.V., 2003.
be even aromas of brandy - in the case of 2- Romanian vineyards and wines, II, Romanian
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The fact that the Sauvignon Blanc variety is Country, AdLibri Publishing House, Bucharest, 57.
also characterized by herbal aromas was also Grigorică L.G., Niculaua M., Nechita C.B., Nistor A.M.,
proved by the presence of compounds that can Cotea V.V., 2017. The impact of some commercial
give this kind of aroma - 3-hexenol acetate, 1- yeast strains on aroma compounds and sensorial
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hexanol, 3-methylbutyl decanoate. In most of Dealurile Olteniei, Romania, BIO Web Conf., 9,
these cases the samples which have a higher 02006.
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Jones G.V., 2007. Climate Change: Observations,
alcoholic concentration. The fatty acids and Projections, and General Implications for Viticulture
their esters have a less pleasant contribution, and Wine Production, Climate and Viticulture
but taking into account the relatively small Congress sponsored by OIV, Zaragoza, Spain, April
concentrations in which they are present it is 10-14.
not disturbing. Most of them are responsible for OIV, 2016. Compendium of International Methods of
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hexanoic acid, isobutanoic acid, ethyl Old Marnie, 2013. Wine: A Tasting Course, Dorling
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class of chemical compounds from which they of the National Meteorological Administration,
come, one can notice that most of the pleasant, Bucharest.
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soil - the basic element of the “Terroir” concept, Our
The perception of an aroma largely depends on terra Publishing House, Iasi.
van Leeuwen C., Darriet P., 2016. The Impact of Climate
the experience and the sensitivity of the Change on Viticulture and Wine Quality, 11, 1,
evaluator. However, the volatile chemical Journal of Wine Economics, 150-167.
compounds which are responsible of aromas Wu Yusen, Duan Shuyan, Zhao Liping, Gao Zhen, Luo
could be better highlighted if there is an Meng, Song Shiren, Xu Wenping, Zhang Caixi, Ma
alcoholic concentration which increases their Chao, Wang Shiping, 2016. Aroma characterization
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Vegetable
Growing
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338
HERITABILITY OF THE TOMATO GENOTYPES RESISTANCE

TO THE HIGH AIR TEMPERATURES
Nadejda MIHNEA, Galina LUPAȘCU
Institute of Genetics, Physiology and Plant Protection, Academy of Sciences of Moldova,

20 Pădurii Street, MD-2002, Chișinău, Republic of Moldova, email: [email protected]
Abstract
Since the resistance of plants to stressful temperatures is a quantitatively controlled polygenetic feature, the
manifestation and particularities of its inheritance in the system of the other quantitative characters, which usually have
correlative relationships, are of interest. The aim of the research was to evaluate the heritability of the tomato
genotypes resistance to the heat and to highlight the most prospects for use in breeding programs. Based on the growth
capacity of the P1 and F1 hybrids, at optimal (25ºC) and high temperature (43ºC), it was found that at the sporophytic
level, the resistance of tomatoes at high temperature is controlled by dominant factors, with varying degrees of
expression and orientation (+/-) that most directly depend on the hybridization components. By calculating the multiple
regression coefficient β regarding the contribution of the maternal and paternal form in the formation of the resistance
of high temperature tomato hybrids F1 (+ 43°C), it was found that the analyzed parameter for the maternal parent is ~
2 times higher than that of the paternal form, which reveals that resistance to heat stress in the case of tomato sporofite
is inherited in particular by the maternal way.
Key words: tomato, heritability, resistance, high temperature.
INTRODUCTION quality at many crop plants, including

leguminous crops (Boote et al., 2005).
Lately, the effects of high temperature, known According to some opinions, heat stress can
as thermal shock, exceed the specific level of also be considered as the temperature level of
normal temperature for agricultural plant 28-29°C which is only a few degrees higher
species, including tomatoes, for which reason than the optimal range of 21-24°C. Increasing
they have become a topical topic for the temperature to such a level does not lead to
fundamental and applied research (Hazra et al., severe disturbances of the biochemical
2007; Mihnea et al., 2011). reactions in the plant that would affect the cell's
More and more obvious and alarming climate functionality, the plants still developing
changes in the world, which frequently cause normally, but the reduction in the number of
decline at crop yields, including tomatoes, have fruits is the general response to this factor, the
led to increased efforts to increase resistance in main cause being the various distortions of the
the breeding programs (Hazra P. et al., 2007). reproductive processes.
Although the tomatoes exhibit a high The high temperature (32/26ºC - day/night),
adaptability to environmental conditions, the and in particular its association with drought,
heat stress can become a major limiting factor greatly affects the growth, development and
for the growth, reproduction and plant productivity of tomatoes. This situation
production level, the optimal temperature for requires the orientation of tomato improvement
cultivation of tomatoes being framed within the programs towards the creation of heat-tolerant
limits of 25-30ºC/20ºC - day/night (Camejo et varieties (Venema et al., 2005; Hazra et al.,
al., 2005). 2007). Increasing the temperature even at 2-4ºC
Most crop plants are subject to high above the optimal level negatively influences
temperature stress (HTS) at certain stages of the development of gametes and inhibits the
development, this phenomenon being more and ability of pollinated flowers to form fruit, thus
more frequently lately (Sato et al., 2000). Plant significantly reducing the yield (Firon et al.,
exposured to HTS reduces harvesting and its 2006).
339
At higher temperatures - than 35°C, germi- Independent variables can correlate with each
nation, flowering, meiotic processes, large fruit other and this is to be taken into account when
formation, egg development, its viability, and determining regression coefficients to avoid
embryo development (Wahid et al., 2007) are misinterpretations caused by false correlations.
affected to the majority of tomato varieties, and The mathematical equation of multiple
resistant tomato genotypes have the ability to regression is as follows:
form a much greater number of fruits than
those sensitive under these conditions у = β 1 • х1 + β 2 • х2+ ... + β n • хn + а,
(Comlekcioglu et al., 2010).
where: y - dependent variable; β - the
MATERIALS AND METHODS regression coefficient for each independent
variable (x1, x2 ... xn); a - constant.
As a research material they have served After evaluating criterion (test) F, it is
varieties of different ecological and important to estimate the regression coefficient
geographical origin and their F1 hybrids. β which can be positive or negative, significant
High temperature reaction testing was carried or insignificant. If the β coefficient is positive,
out according to the method proposed by Ivakin it can be interpreted that for each increase of
(1979), based on plant growth capacity after the predictor variable with 1 unit, the increase
maintaining it at elevated temperatures for 6 of the dependent variable will be equal to the
hours. For the analysis of the high temperature value of the non-standardized coefficient β
influence on the germination and the length of (Nathans et al., 2012).
the plants, the thermal level of 430C was used. The data obtained were statistically processed
According to the method, between the in the software package STATISTICA 7.
resistance of the sporophyte and the tomato
gametophyte there is a positive correlation. RESULTS AND DISCUSSIONS
The degree of domination (hp) was calculated
based on the formula (Briubeiker, 1966): One of the basic genetic indices that
hp = F1 – 0.5 (P1+P2)/ Hp – 0.5 (P1+P2), in demonstrates the type of inheritance in F1
which: F1 - the mean value of the character in generations is the degree of domination (hp). To
the F1 generation; P1, P2 - the mean value of the obtain F1 hybrids, crossbreeds were made
character in the parental forms; Hp - average between parental forms with varying degrees of
value of character rated at best parental resistance / sensitivity to high temperatures.
component. The obtained data on the reaction of high-
Clusterian analyzes were performed by temperature hybrids of tomato populations
constructing dendrograms based on the Ward show that hybrid combinations F1 have values
method and the k-means method (Savary et al., that show increased resistance of the obtained
2010). In the k-means method, 3 clusters were hybrids compared to the genitors, in some
programmed after the possible values of the combinations the F1 values are lower than the
characters: small, medium and high. The main average of the parents. The analysis of the
purpose of these procedures is to find the genitors and F1 hybrids resistance at t = 42-
similarities and differences between objects 430C showed a considerable variability of the
(genotypes) according to the parameters used studied character: 23.6 ... 56.0% (Table 1).
and their distribution in groups so that objects The degree of resistance of high temperature
in the same group are similar and those in plants was high to the hybrids F1: Visas x
different groups - special. Sunmark, Katerina x Sunmark, A 90/7 x Gusar,
Multiple regression analysis. The overall Katerina x Danna, the values of which
importance of this analysis is to elucidate the constituted 57.5, 53.6, 52.6, 52.2%,
links between several (or more) independent respectively.
variables and a dependent variable.
340
Table 1. Heritability of the resistance/sensitivity of tomato genotypes at high temperature at the juvenile plant level
Nr. Combination Resistance of the plant to t=430C, %
P1 P2 F1 hp
1. Onix x Saladette 28.9 31.3 33.6 2.92
2. Katerina x Burnley Metro 41.8 38.1 35.5 -2.31
3. A 90/7 x Gusar 38.6 20.4 52.6 2.54
4. Narvic x Zastava 34.3 56.0 39.4 -0.53
5. Cal J THM х Saladette 28.1 31.3 42.7 8.12
6. Katerina x Zastava 41.8 56.0 49.6 0.09
7. Nistru x Оnix 30.9 28.9 40.9 11.0
8. A 90/7 x Кredo 38.6 35.3 46.9 5.9
9. Viza x Burnley Metro 38.5 38.1 23.6 -73.5
10. Katerina x Danna 41.8 40.0 52.2 12.5
11. Katerina x Sunmark 38.6 47.2 53.6 2.5
12. A 90/7 x Costral 30.9 31.3 46.2 75.5
13. Nistru x Saladette 38.5 64.0 46.4 -0.37
14. Viza x Sunmark 41.8 64.0 57.5 -0.4
The appreciation of the high temperature x Credo, A 90/7 x Costral, Katerina x Sunmark,
influence on the growth of tomato plants has Nistru x Saladette, of the 14 studied.
revealed considerable variability in both Dominance of sensitivity was recorded at the
parental and hybrid combinations ranging from A 90/7 x Gusar combinations, and the
2.2 ... 5.5 cm and 2.4 ... 5.8 cm, respectively intermediate sensitivity inheritance at Viza x
(Figure 1). Burnley Metro.
At high temperatures, F1 hybrids showed Considering that the phenomenon of
advanced positive resistance compared to the overdominance is not inherited (because it is a
best genitor at the combinations: Onix x result of allelic interactions), we can deduce
Saladette, A 90/7 x Gusar, Cal J THM x that in the descendants of the combinations in
Saladette, A 90/7 x Kredo, Katerina x Danna, A which F1 manifested resistance much higher
90/7 х Соstral, Nistru x Onix. than that of the best parent, more probably will
The values of the hp coefficient for the length not be found genotypes with this level of
of the plantlets reveal the overdominance of the character manifestation.
resistance to 11 combinations: Onix x The success of the selections will be
Saladette, Katerina x Burnley Metro, Narvic x determined only by the possibility of
Zastava, Cal J THM x Saladette, Katerina x identifying homozygous resistance forms.
Zastava, Nistru x Onix, A 90/7 x Kredo, A 90/7
cm
6
0
1 2 3 4 5 6 7 8 9 10 11 12 13 14
The maternal form The paternal form
The hybrid combination
Figure 1. Comparative data of growing parental formulas and F1 hybrids at high temperature (43°C):
1 - Onix x Saladette; 2 - Katerina x Burnley Metrou; 3 - A 90/7 x Gusar; 4 - Narvic x Zastava; 5 - Cal J THM x
Saladette; 6 - Katerina x Zastava; 7 - Nistru x Onix; 8 - A 90/7 x Kredo; 9 - Viza x Burnley Metrou; 10 - Katerina x
Danna; 11 - Katerina x Sunmarк; 12 - A 90/7 x Costral; 13 - Nistru x Saladette; 14 - Viza x Sunmark
341
By cluster analysis (A) and multidimensional plantlets at + 43°C, forming clusters at different
scaling (B), it was found that the investigated levels of aggregation or different location in the
combinations differ greatly from the reaction of three-dimensional space (Figure 2).
W ard`s method
Euclidean distances
1
5
7
12
3
2
9
8
10
11
4
6
13
14
0 20 40 60 80 100
Linkage Distance
Germination Length of radicle, mm

Figure 2. The dendrogram for the distribution of tomato combinations based on the reaction
of parent plants (P1, P2) and high temperature hybrids F1
The k-means clustering analysis of the showed the paternal form of the combination
combinations studied (based on the parental (Table 2, Figure 3).
and hybrid F1 reactions) in 3 clusters according Thus, the intercluster variance determined by
to the possible values of the length of the the parental form (P2) recorded the highest
plantula - large, medium, small, highlighted value: 2149.821.
that the greatest differentiation capacity they
Table 2. Analysis of cluster variance

Genotype Intercluster variance df Intracluster variance df F p
P1 210.959 2 118.196 11 9.817 0.0004
P2 2149.821 2 295.512 11 40.012 0.0001
F1 221.282 2 872.150 11 1.396 0.2883
*- p≤0,05.
%
70
65
60
55
50
45
40
35
30 1
2
25 3
20
P1 P2 F1
Figure 3. The ability to differentiate clusters (k-means method) by parental forms and tomato hybrid F1
It should be noted that by calculating the hybrid F1; P1 - maternal form; P2 - paternal
multiple regression β coefficient regarding of form. As it can be seen, the β coefficient in the
the maternal and paternal form contribution in case of the maternal is ~ 2 times higher than
the formation of the phenotype of hybrid F1 that of the paternal parent, which shows that the
plants, the following regression equation was high temperature resistance in tomato plants is
found: y = 0.289 P1 + 0.139 P2, in which: y - inherited in particular by the mother.
342
CONCLUSIONS Agong S.G., Schittenhelm S., Friedt W., 2000. Genotipic
variation of Kenyan tomato (Lycopersicon esculentum
L.) germoplasm. In: Plant Gen. Res. Newsl, 61-67.
Based on the growth capacity of the P1 and F1 Boote K.J. et al., 2005. Elevated temperature and CO2
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(43ºC), it was found that at the sporophytic several globally important crops. J. Agric. Meteorol of
level, the resistance of tomatoes at high Japan. 60 (5): 469-474.
temperature is controlled by dominant factors, Camejo D. et al., 2005. High temperature effects on
photosynthetic activity of two tomato cultivars with
with degrees variation of expression and different heat susceptibility. J. Plant Physiol., 162
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By calculating the multiple regression heat tolerant tomato (Solanum lycopersicon) genotypes
coefficient β regarding of the maternal and by SRAP and RAPD markers. Genet. and Mol. Res., 9
(4): 2263-2274.
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Nathans L.L., Oswald F.L., Nimon K., 2012. Interpreting
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343
344
ESTIMATION OF VARIABILITY PARAMETERS OF SOME QUALITATIVES

COMPONENTS AT A SET OF SWEET CORN LINES FROM
AGRICULTURAL RESEARCH AND DEVELOPMENT STATION TURDA
Luana PĂCURAR (GRECU)1, Alexandru Ioan APAHIDEAN2, Gheorghiţa HOZA4,

Maria DINU3, Rodica SOARE4, Maria APAHIDEAN2, Voichiţa HAŞ1
1
Agricultural Research and Development Station Turda (SCDA Turda), 27 Agriculturii,
Street, Turda, Cluj, Romania
2
University of Agricultural Sciences And Veterinary Medicine Cluj-Napoca,
3-5 Calea Mănăștur, Cluj-Napoca, Romania
3
University of Craiova, Faculty of Horticulture, 19 Libertății Street, Craiova, Romania
4
Abstract
Sweet corn is a recent crop in Romanian agriculture, being cultivated more and more. Modern sweet corn, like all
crops, is the product of its own evolution, in which human intervention was the first unconscious phase, becoming
conscious over last two centuries. Knowledge of this development may be useful for forming the basis for planning for
future improvement of this plant. Purpose of this study was to analyze inbred maize lines from germplasm collection
from Turda Agricultural Research Center in terms of variability of some chemical components. Biological material was
represented by 27 inbred lines of sugar-1 gene. Each variant was sown on four rows, an average of 40,000 pl/ha.
Quality analyzes were performed using TANGO NIR spectro-photometer. Chemical parameters analysed were: protein
content, starch, sugar, NDF (neutral detergent fibers) and NCGD (neutral cellulose digestibility). Between analyzed
lines there is an important variability of sugar content.
Key words: lines, sugar, starch, sweet corn, variance.
INTRODUCTION Sweet corn is one of the most recent crops in

the landscape of Romanian agriculture, being
Globally, sweet corn represents 7.7% of total cultivated more and more. Most of sweet corn
vegetable consumption and ranks 2nd after production from Romania is obtained in the
tomatoes for US and Canada consumers western part of the country and is not processed
(McMullen et al., 2009). Due to the high in the country, but taken to Hungary
content of carbonates and other nutrients such (www.agrointel.ro).
as vitamins C, A, B1, B2, proteins, fats, starch, At Turda Agricultural Research Station, in
cellulose and mineral components (potassium, 1971, a genetic improvement program was
magnesium, calcium, phosphorus, iron, launched to obtain sweet corn on the basis of
selenium, copper, nickel and chrom) and infusion of su1 recessive gene to a series of
different uses for both fresh consumption and normal inbred lines. The obtained sugar lines
food industry, this species was quickly adopted were studied in a diaphragm crossing system,
worldwide (Hefny, 2011). obtaining the general and specific combination
Recently, in U.S., use of high fructose syrup capacity for the specific characters of sweet
obtained from sweet corn and used as a corn. Based on the behavior of studied inbred
sweetener gained a large market share (Revilla lines, the formula of DULCIN tri-linear hybrid
et al., 2005). Sweeteners made from sweet was approved in 1988 (Căbulea et al., 1994;
corn, belong to the best sweeteners class in US Has V., 2000).
with a 52% market share due to their stability, Over past 15 years, Turda Agricultural
high functionality and ease of use (Singh et al., Research Center has developed an intensive
2014). program to obtain new, early-quality inbred
345
lines with superior qualities, which has led to of variability of some chemical components
recording of new genetic advances in (sugar, starch, protein, NDF and NCGD) was
amelioration of sweet corn. proposed.
As a result of using initial material from very
early American and Canadian origins, the pa- MATERIALS AND METHODS
rental forms of PRIMA hybrid were obtained,
approved in 1991 and considered the earliest Biological material was represented by 27
hybrid from sweet corn hybrids experimented inbred lines of sugary-1 gene. Each variant was
in CIOS network (Has et al., 1994). sown on four rows of 5 m in length and at a
Modern sweet corn, like all crops, is the distance of 0.70 m corresponding to an average
product of its own evolution, in which human of 40,000 plants/ha. This group of lines was
intervention was the first unconscious phase, subjected to quality analysis, which was
becoming conscious over last two centuries performed with the TANGO NIR
(Srdić et al., 2012). Knowledge of this spectrophotometer (Bruker Optik Gmbh,
development may be useful for forming the Ettingen, Germany).
basis for planning for future improvement of From each line, self-pollinated cobs were
this plant (Lazcano et al., 2011). selected to eliminate effects of alopolen sources
It is important to know basic materials involvement in the analyzed chemical
(varieties, populations) from which modern components. Sweet corn cobs were analyzed at
inbred lines have been extracted to retrieve lost full maturity, being allowed to lose their
characters during inbreeding (Kashiani et al., moisture by natural drying in laboratory, after
2010). Sweet corn breeders have paid attention which grain samples were taken from each line.
to varieties, both as starting material for inbred For each line of self-pollinated corn cobs that
lines and as new sources of favorable alleles have been harvested, samples of about 50
(Huelsen, 1954; Tracy, 1994). The one who grams of grains were harvested, which were
dealt with study of varieties, their origins, role subsequently ground using a WZ-1 knife mill
played in sweet corn evolution of was Walton (Sandkiewicz Instruments - Poland), and then
C. Galinat (1971) in the study: "The Evolution sifted with a sieve.
of Sweet Corn". Chemical parameters analyzed were: protein
Over the last 50 years, production gain in sweet content, starch, sugar, NDF (neutral detergent
corn, attributed to genetic gain, was modest fibers) and NCGD (neutral cellulose
compared to that obtained with ordinary corn digestibility).
(Letrat and Pulam, 2007). Important reasons for Based on variance analysis, some derived
this could be: genetic parameters were also calculated:
- narrow genetic basis of sweet corn which,
according to some authors, is largely derived ‫ܩ‬ሺ‫݁ܿ݊ܽ݅ݎܽݒܿ݅ݐ݁݊݁ܩ‬ሻ
from Northern Flint and which in its evolution െ ሺሻ
has gone towards a severe narrowing of ൌ
ሺሻ
variability;
- non-Northern Flint germplasm, represented ܸ‫ܧ‬ሺ݁݊‫݁ܿ݊ܽ݅ݎܽݒݐ݊݁݉݋ݎ݅ݒ‬ሻ ൌresidual variance;
୚୑
by Stowell's Evergreen and County Gentleman ܸܲሺ‫݁ܿ݊ܽ݅ݎܽݒܿ݅݌ݕݐ݋݄݊݁݌‬ሻ ൌ ܸ‫; ܩ‬
ୖ
varieties, emerged through the introgression of ξ௏ீ
CVG% (Coefficient of genetic variationሻ ൌ x100;
sugary-1 gene in germplasm dentiformis ଡ଼
(Revilla et al., 2009); CVP% (Coefficient of phenotypic variation) =
ξ௏௉
x100;
- excessive use of "Golden Bantam" type in ଡ଼
CVM% (Coefficient of environment variation) =
creation of inbred lines (as a source of yellow ξ௏ெ
endosperm and taste qualities). Current germ x100
ଡ଼
cell gene mutation contains, to a greater or (Singh and Chaudhary, 1985);
lesser extent, genes of Golden Bantam genus in ௏ீ
Hሺሻ ൌ
pedigree (Romay et al., 2013). ௏௉
Following this study, analysis of inbred maize (Falconer, 1971);

lines from Turda germplasm collection in terms X = character average.
346
Generated results will be chemometrically improvement program if they are valuable
processed using main component analysis and under other aspects (cob production, precocity,
cluster analysis; such charts can provide cob size uniformity etc.).
valuable information about possible corre- Analyzing data in Table 2 led to the conclusion
lations between studied variables or samples that there are considerable differences between
composition, especially when working with analyzed lines with respect to sugar content,
large sets of values. Also, resulting clusters having highest C.V. of 39.2%, which suggests
provide valuable insights into possible simila- the existence of a great variability of this im-
rities between lines and also their grouping portant taste characteristic. As a matter of fact,
according to these similarities or differences. wide range of minimum and maximum values
indicates a significant variation in sugar content.
Table 2. Variability parameters of protein content,
starch, NCGD sugar and NDF at a set
Chemical composition of maize lines studied is of sweet corn lines (Turda, 2016)
shown in Table 1.
Indicators CV Mini- Maxi- Average Standard
Table 1. Chemical composition of a set (%) mum mum deviation
of sweet corn lines (Turda, 2016) Protein 17.4 9.3 19.88 12.70 2.21
Sugar 39.2 0.28 6.70 4.3 1.69
Lines NCGD NDF Protein Starch Sugar Starch 14.5 25.72 54.01 46.95 6.83
(%) (%) (%) (%) (%) NDF 12.5 11.73 19.17 14.76 1.85
Ta su 21Q206 86.18 14.26 11.33 49.68 5.67 NCGD 1.6 83.76 88.94 86.75 1.4
TA su 22 87.95 12.09 11.78 47.66 4.83
Tsu 152 88.07 15.55 13.26 49.94 6.67
Tsu 345R 86.67 16.71 12.09 45.83 6.70 CV% values reflect moderate variability within
TA 26 87.75 13.35 12.07 52.73 3.72 the group of lines analyzed for protein and
TA 27 87.79 12.51 12.53 53.65 2.16 starch content. Differences between minimum
TA 28 87.41 14.19 11.64 47.54 2.83 and maximum values of protein and starch
TD 101 88.66 15.80 9.30 53.25 6.52 content indicate an important variability of
TD 102 87.39 14.18 12.47 46.81 4.40 these components and the possibility of
TA 28 87.11 15.38 12.52 50.62 3.73
TD 110 85.68 14.07 14.43 49.77 1.41
selecting valuable genres both as variants and
TVa 638 87.73 15.14 12.58 49.53 2.72 as minus variants according to aims pursued by
Tva 642 87.66 15.53 11.08 51.57 1.92 the breeder. Wide variation limits of the two
TVa 647 87.59 13.06 13.63 49.55 0.28 components show existence of significant
TV a 670 87.76 16.63 13.25 46.51 5.04 variations in the group of analyzed lines.
T su 209 84.64 19.17 12.41 44.88 5.26 Lowest variability is recorded at neutral
T Su 233 86.54 14.89 13.43 45.83 4.07 cellulose digestibility (NCGD) values. Reduced
T Su 244 85.35 17.84 12.06 38.94 6.03
P 51 84.06 14.95 12.80 44.73 6.66
values of CV indicates a small fluctuation of
LC 154/74 85.14 16.35 11.53 47.29 6.33 these values around average. Therefore, we can
SW 87 83.76 16.07 14.80 42.31 4.85 say that there are no large differences in
TC 179 su 86.86 12.08 12.48 49.59 4.00 digestibility of cellulose between analyzed
TD 111 su 87.93 12.95 10.66 50.86 4.78 lines. This parameter is mainly used in fodder,
TD 112 su 88.94 11,73 10.41 54.01 3.89 for determination of formulas used for
TD 113 su 85.75 15.15 18.19 27.44 4.01
calculating of fodder energy values.
TD 114 su 84.81 16.40 19.88 25.73 4.09
TD 115 su 87.24 12.52 10.31 51.50 3.68 A moderate variability of studied parameters
can also be noticed for neutral detergent fibers
Higher sugar content was found in Tsu 152 and (NDF) represented by fractions of insoluble
Tsu 345R lines, which can indicate them as cell walls (hemicelluloses, celluloses, lignin,
possible genres (only after being tested for tannins etc.).
combining ability) in amelioration of sugar Generally, the two parameters, NDF and
content in sweet corn lines with a high yield but NCGD, are used to characterize food
a lower sugar content. On the opposite side are digestibility.
Tva 642, TVa 647 and TD 110, which probably With higher protein content, lines in the
should be included in a sugar content HONEY group (TD 113 su and TD 114 su)
347
stand out, with 19.88% protein and 19.8% probably because of lower number of analyzed
respectively, as can be seen in Figure 1. Also cases.
known is the reverse relationship of sugar and Large sample dispersion around the regression
protein, which is confirmed by this study, as line suggests that there is insignificant
protein content is reduced when sugar content interaction between the two variables and that
is increased, with deviations (Figure 1). choosing forms with lower starch content is not
Between the two important chemical necessarily associated with a significant
components of sugar and starch there is a well- increase in sugar. A significant deviation that
established logical correlation even though it is strengthens this negative relationship can be
not statistically assured in present case, noticed within lines of the Honey group within
the circle in Figure 2.
25,00 PROTEINE
Protein
Sugar
ZAHAR
20,00
15,00
10,00
5,00
0,00
TD 110
Ta su 21Q
T su152
Tva 642
T su 209
LC 154/74
TC 179 su
T su 345R
TD 101
TD 102
TVa 638
TV a 647
T Su 233
T Su 244
P 51
TD 111 su
TD 112 su
TD 113 su
TD 114 su
TD 115 su
TA su 22
TA 26
TA 27
TA 28
TA 28
TV a 670
SW 87
Figure 1. Protein and sugar content of 27 sweet corn lines (Turda, 2016)
8,00
R² = 0.027
7,00
r= -0,16 n.s
6,00
Sugar content (%)
5,00
4,00
3,00
2,00
1,00
0,00
15,00 25,00 35,00 45,00 55,00 65,00
Starch content %
Figure 2. Correlation between sugar and starch content
Analyzing obtained data, between the analyzed quality heterosis superior to parental forms.
lines there is an important variability of sugar These lines are therefore of real use in
content, values of sample F being very improving the taste of sweet corn. Sugar
significant (Table 3). It can be stated with content of sweet corn lines that directly
certainty that at the level of lines one can find influences their quality must be correlated with
some forms that included in hybridization other factors such as cob production but also
programs can lead to the expression of a higher through other selection criteria.
348
Table 3. Analysis of the variance for sugar content Improving chemical composition of graines
in the 27 sweet corn lines requires the combination of a set of procedures
Variability SPA GL S2 F based on reciprocal recurrent selection, tandem
source selection with the requirement to control
G 216.22 26 8.32 34.74*** expression under various environmental
R 7.03 2 3.52 conditions.
Error G 12.45 52 0.24
From the statistical analysis of the weight of
the factors involved in the expression of the
Based on proposed calculation formulas from
grain composition, contribution of genotype to
Singh (2014), genetic variation coefficients,
environmental factors is predominant. Major
environmental (residual) and phenotypic varia-
share of genotype contribution in expressing
tion coefficients as well as broader heritability
grain quality suggests that sweet corn grain
were calculated (after Nordby, 2008).
lines have relatively stable components in
Analyzing data from Table 4, high values of
relation to environmental conditions.
genetic variation indicate the important
contribution of genetic factor in obtaining sugar
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variance, suggesting major implications of the porumb cu bobul zaharat creați la Stațiunea de
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sugar content that can be improved by Gerdens J.T., Tracy W.F., 1994, Diversity of historically
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Similar result were obtained by Haş and Haş in Sci 34: 26-33.
their study in 2009. Has V., Căbulea I., Grecu C., Haș I., Marușca P., 1994.
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Contribuții ale cercetării șt. la dezv. agric. vol. V:
Table 4. Estimators of sugar content variability in 143-152.
analyzed sweet corn lines Haş V., 2000. Research regarding to the inheritance of
Genetic Enviro- Pheno- Coeffici- Coefficient Coefficient some qualitative and quantitative characters in sweet
Varian- ment typic ent of of H2 corn Ph. D.
ce variance variance of genetic environ- phenotypic Haş V., Haş I., 2009. Genetic Inheritance of Some
VG VM VP variation ment variation Important Characters of Sweet Corn, Notulae
CVG % variation CVP% Botanicae Horti Agrobotanici Cluj-Napoca; Cluj-
CVM% Napoca Vol. 37, Iss. 1, 244-248.
Sugar content Hefny M., 2011. Genetic Parameters and Path Analysis
1.6 3.52 2.77 29.41 43.63 38.71 0.57
of Yield and its Components in Corn Inbred Lines
(Zea mays L.) at Different Sowing Dates, Asian
CONCLUSIONS Journal of Crop Science, 106-117.
Huelsen W.A., 1954. Sweet corn. Interscience Publisher,
Use of sugary-1 (su-1) mutant gene for New York.
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with complex determinism, both due to Lazcano C., Revilla P., Malvar R.A., Domínguez J.,
interactions between quality gene (su-1) of the 2011. Yield and fruit quality of four sweet corn
endosperm and the polygenic complex on hybrids (Zea mays) under conventional and integrated
which it operates, as well as environmental fertilization with vermicompost. Journal of the
Science of Food and Agriculture, Volume 91, Issue 7,
conditions. 1244-1253.
Higher heritability of transmitting sugar content Lertrat K., Pulam T., 2007. Breeding for increased
makes recurrent selection during inbreeding sweetness in sweet corn, Int J Plant Breed, 27- 29.
very effective. Thus, improvement and fixation McMullen M.D., Kresovich S., Villeda H.S., Bradbury
by individual selection of the quality character P., Li H., Sun Q., Flint-Garcia S., Thornsberry J.,
Acharya C., Bottoms C., Brown P., Browne C., Eller
is very effective in the course of inbreeding.
349
M., Guill K., Harjes C., Kroon D., Lepak N., performance of maize plants carrying the allele
Mitchell S.E., Peterson B., Pressoir G., Romero S., sugary1, Plant Breeding, 92-95.
Rosas M.O., Salvo S., Yates H., Hanson M., Jones Romay M.C., Millard M.J., Glaubitz J.C., Peiffer J.A.,
E., Smith S., Glaubitz J.C., Goodman M., Ware D., Swarts K.L., Casstevens T.M., Elshire R.J., Acharya
Holland J.B., Buckler E.S., 2009. Genetic Properties C.A., Mitchell S.E., Flint-Garcia S.A., McMullen
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*** www.agrointel.ro.
350
RESEARCH REGARDING THE INFLUENCE OF FERTILISER RATES ON

MORPHOLOGICAL FEATURES IN COMMON BEAN (PHASEOLUS
VULGARIS L. CONVAR. VULGARIS) PODS CULTIVATED IN SOLARIUM
AT THE DIDACTIC AND RESEARCH BASE IN TIMIŞOARA, ROMANIA
Marius BALINT, Alin DOBREI, Alina Georgeta DOBREI, Gheorghe POȘTA
Banat University of Agricultural Sciences and Veterinary Medicine „King Michael I of Romania”
from Timisoara, 119 Aradului Street, Timisoara. Romania
Abstract
Common bean or green bean (Phaseolus vulgaris L. - Family Papilionaceae syn. Fabaceae) is cultivated for its young
pods before seed formation, used in some dishes and in the preserve industry. Trial was conducted in the solarium of
the Didactic and Research Base in Timisoara, Romania, on 400 m2. It was a poly-factorial trial with sub-divided plots
and three replicates: Factor A (supplementary fertilisation) with 3 graduations (a1 - Cropmax; a2 - Lithovit; a3 -
Trainer); Factor B (basic fertilisation) with 3 graduations (b1 - Orgevit; b2 - Phenix; b3 - Italpollina) and Factor C
(cultivar) with 3 graduations (c1 - Goldfield; c2 - Ecaterina; c3 - Aurie de Bacău). As far as pod length is concerned,
there are distinctly significantly positive differences (1.54-1.70 cm) between the cultivar Ecaterina and the trial mean.
As for pod width. there are very significantly positive differences (0.05-0.15 cm) between the three supplementary foliar
fertilisers. From the perspective of pod diameter, there are very significantly positive differences (0.01-0.05 cm) also
between the three supplementary foliar fertilisers.
Key words: Phaseolus vulgaris L. convar. vulgaris, common bean, morphological characters, fertiliser rates, solarium
type.
INTRODUCTION The stem is cylindrical-edged; it is covered by

hairs. ramified in dwarf common bean
Common bean or green bean (Phaseolus cultivars, 25-30 cm tall, and voluble; it is less
vulgaris L. - Family Papilionaceae syn. ramified in climbing common bean cultivars,
Fabaceae) is cultivated for its young pods and 4-5 m tall (Poşta, 2008).
harvested before seed formation that are used in Common bean cultivars with undetermined
meals and in the can industry (Apahidean and growth bloom 60-70 days after sprouting and
Apahidean, 2000; Broughton et al., 2003). continue to bloom.
The species originates in Central America and The fruit is a dehiscent pod characteristic in
South America (Peru, Mexico), where Aztec size, shape and colour depending on the
tribes used to cultivate it in times immemorial. common bean cultivar.
It was brought to Europe in the 16th century by As a plant originating from warm areas, it is
the Spaniards and the Portuguese (Indrea et al., heat demanding. Seed minimum germination
2007). It spread from Spain and Portugal to the temperature is 15°C, when sprouting occurs in
Netherlands, France, Germany and England 12-14 days. Optimum temperature is 20-27°C,
(Ciofu et al., 2004). when sprouting occurs in 4-8 days. During
Common bean is an annual. herbaceous. vegetation, optimum temperature is 20-25°C,
thermophilous plant. Two convarieties are while minimum temperature is 10-12°C
cultivated: nanus, covering plants with (Konsens et al., 1991). Below 10°C growth
determined growth, and vulgaris, covering stops and at -0.5°C, the plants die.
plants with undetermined growth (Poşta, 2008). Common bean is a short-day plant. With long-
The root system is superficial and it goes 30-40 day conditions, vegetative growth is intense.
cm deep in the soil. Some toots even reach 100 detrimental to fructification, which asks, in
cm deep in the soil. The roots have nodosities climbing common bean cultivars, the removal
containing nitrogen-fixing bacteria. of leaves and branches once a week to favour
351
larger numbers of flowers and, implicitly, of Foliar organic fertiliser Trainer (Italpollina spa.
pods. Critical water phases are when seeds Italy) contains organic nitrogen (5%), vegetal
germinate, at blooming and at pod formation amino acids (26.3%) and organic matter
(Beebe et al., 2013). Lack of soil water hinders (35.5%). Application concentration is 0.3-
blooming, while low air moisture causes flower 0.4%.
abortion. Common bean responds well to Certified ecological organic fertiliser Orgevit
phosphorus and potassium fertilisation. On contains N (4.0%), P2O5 (2.5%), K2O (2.3%),
poor soils, it is recommended to apply nitrogen Ca (9.3%), MgO (1.1%), organic substances
fertilisers during the first vegetation phases (65% guano) and microelements (Fe, Mn, Bo,
(Araújo et al., 2004; Soratto et al., 2010; Zn, Cu, Mo). In vegetables cultivated in
Turuko and Mohammed, 2014). greenhouses and solaria. application rate is 1.5-
2.0 t/ha.
MATERIALS AND METHODS Phenix is an organic fertilizer containing N
(6.0%), P2O5 (8.0%), K2O (15.0%), MgO
Experiments were carried out at the (3.0%), organic carbon (29.0%) and organic
Experimental Station of the Faculty of substances (50.0%). Application rate in
Horticulture and Forestry of Timisoara in a 400 greenhouses and solaria is 5.0 t/ha.
m2 solarium, built and equipped with Natural organic fertiliser Italpollina (Italpollina
environmental automated control systems and spa. Italy) used to fertilise the soil contains
equipments. 4.0% N, 4.0% P2O5, 4.0% K2O, 0.5% MgO and
Experiments carried out during 2016-2017 had 41% C (organic carbon). Application rate
a polyfactorial character; variants were set after homologated in vegetables cultivated in
the randomised block method with three greenhouses and solaria is 3-4 t/ha.
replicates specific to experiments in forced The biological material used in the trials
protected areas of vegetable culture. consisted in nursery seedlings sown directly in
Factor A (supplementary fertilisation) with 3 9 cm diameter pots; seedling age upon planting
graduations: a1 - Cropmax; a2 - Lithovit; a3 - was 25-30 days. The planting scheme was on
Trainer. 90 cm equidistant rows, while planting distance
Factor B (basic fertilisation) with 3 gra- per row was 35-40 cm.
duations: b1 - Orgevit; b2 - Phenix; b3 - During vegetation, we monitored the soil to
Italpollina. determine the variability of the main soil
Factor C (cultivar) with 3 graduations: c1 - morphological features.
Goldfield; c2 - Ecaterina; c3 - Aurie de Bacău. Observations were made using the current
The growth bio stimulator Cropmax (Holland technique of descriptors and evaluation grades
Farming B.V., Holland) contains amino acids, specific to the species (Ciulca, 2002).
macro- and micro-elements, vitamins and
polysaccharides, being 100% organic. This RESULTS AND DISCUSSIONS
fertiliser contains N (0.2%), P (0.4%), K
(0.02%), Fe (220 mg/l), Mg (550 mg/l), Zn (49 Soils favourable to common bean are rich in
mg/l), Mn (54 mg/l), Cu (35 mg/l), Bo (70 humus and have a neutral to alkaline reaction
mg/l), Ca + Mo + Cb + Ni (10 mg/l), vitamins (pH = 6.5-7.5) (Indrea et al., 2007).
C and E, enzymes and carotenoids. Having a short vegetation period. climbing
Recommended concentration rate in solarium common bean extracts from the soil small
crops is 0.2%, every 7-10 days. amounts of easily assailable nutrients. Common
Foliar fertiliser Lithovit standard (zeovita bean responds well to fertilisation with
GmbH. Roter) contains 75% CaCO3, 4% phosphorus and potassium and. on poor soils, it
MgCO3, 0.25% Fe, 5.0% SiO2, 0.1% K2O, is recommended to apply nitrogen fertilisers
0.015% N, 0.015% P2O5, 0.01% Mn, 0.002% during the first vegetation phases, upon
Cu and 0.005% Zn. This ecological fertiliser is blooming and upon pod formation (Mourice
applied in concentrations of 0.5%, every 15 and Tryphone, 2012). Specific nutrient
days. consumption per ton of fresh produce is 7-9 kg
352
a.s. N, 2.5 kg a.s. P2O5, 6.5-7.0 kg a.s. K2O, 10 the interdependence between basic and
kg CaO (Davidescu and Davidescu, 1992). supplementary fertilisations and genetic factor
Assessing the influence of fertiliser rate on (cultivar). Table 1 below show trial results
some morphological and yielding features in regarding the unilateral influence of
common bean was done from the perspective of supplementary fertilisation on pod length.
Table 1. Influence of supplementary fertilisation on climbing common bean pod length

(Didactic Base Timişoara, 2016-2017)
Supplementary fertilisation Pod length (cm) Relative values (%) Difference/Significance
Cropmax 23.79 103.80 0.87**
Lithovit 22.36 97.60 -0.560
Trainer 22.60 98.60 -0.32
Control (average exp.) 22.92 100.00 0.00
LSD5% = 0.39 cm; LSD 1% = 0.59 cm; LSD 0.1% = 0.94 cm.
Comparative analysis of the three products concentration in macro- and micro-elements of

used in supplementary fertilisation shows the fertiliser Cropmax.
distinctly significant positive differences of Our research also aimed at assessing the
climbing common bean pod length (+0.87 cm) influence of basic fertilisation on climbing
when applying the product Cropmax. This common bean pod length. Trial results are
difference is due to the composition and shown in Table 2 below.
Table 2. Influence of basic fertilisation on climbing common bean pod length

Basic fertilisation Pod length (cm) Relative values (%) Difference/Significance
Orgevit 22.75 99.30 -0.17
Phenix 23.80 103.90 0.88***
Italpollina 22.20 96.90 -0.72000
When using the three basic fertilisers cultivated unit due to its biological and morphological
in solaria, there is a very significant positive features (Poşta and Berar, 2005). Trial data
difference in pod length (+0.88 cm) when regarding the influence of soil on climbing
applying the fertiliser Phenix. common bean pod length are shown in Table 3
The genetic factor (cultivar) used in the trial below.
has a primordial influence on yield per area
Table 3. Influence of cultivar on climbing common bean pod length
Cultivar Pod length (cm) Relative values (%) Difference/Significance
Goldfield 25.33 110.50 2.42***
Ecaterina 22.01 96.10 -0.90000
Aurie de Bacău 21.40 93.40 -1.51000
As for the data shown in Table 3 above, there is For better assessment of the interdependence
a very significant positive difference in pod between trial factors we assessed the
length (+2.42 cm) in the Goldfield climbing combination between basic fertilisation.
common bean. This significant change of the supplementary fertilisation and cultivar. Trial
morphological feature is due to the biological results are shown in Table 4 below.
improved cultivar feature (Madoşă, 2000;
Nedelea and Madoşă, 2004).
353
Table 4. Interdependence of the combination basic fertilisation x supplementary fertilisation x cultivar on climbing
common bean pod length (Didactic Base Timişoara, 2016-2017)
Factor combination Pod length (cm) Relative values (%) Difference/Significance
Orgevit x Cropmax x Goldfield 25.50 111.30 2.59***
Phenix x Cropmax x Goldfield 27.10 118.28 4.19***
Italpollina x Cropmax x Goldfield 26.30 114.79 3.39***
Orgevit x Cropmax x Ecaterina 22.42 97.86 -0.4900
Phenix x Cropmax x Ecaterina 24.39 106.46 1.48**
Italpollina x Cropmax x Ecaterina 21.74 94.89 -1.170
Orgevit x Cropmax x Aurie de Bacău 22.94 100.13 0.03
Phenix x Cropmax x Aurie de Bacău 22.13 96.59 -0.78
Italpollina x Cropmax x Aurie de Bacău 21.59 94.23 -1.320
Orgevit x Lithovit x Goldfield 23.96 104.58 1.05*
Phenix x Lithovit x Goldfield 25.44 111.04 2.53***
Italpollina x Lithovit x Goldfield 24.45 106.72 1.54**
Orgevit x Lithovit x Ecaterina 21.07 91.96 -1.84000
Phenix x Lithovit x Ecaterina 23.17 101.13 0.26
Italpollina x Lithovit x Ecaterina 20.21 88.21 -2.70000
Orgevit x Lithovit x Aurie de Bacău 21.56 94.10 -1.3500
Phenix x Lithovit x Aurie de Bacău 21.02 91.75 -1.89000
Italpollina x Lithovit x Aurie de Bacău 20.07 87.60 -2.84000
Orgevit x Trainer x Goldfield 24.22 105.71 1.31*
Phenix x Trainer x Goldfield 25.31 110.47 2.40***
Italpollina x Trainer x Goldfield 24.72 107.90 1.81***
Orgevit x Trainer x Ecaterina 21.28 92.88 -1.6300
Phenix x Trainer x Ecaterina 23.41 102.18 0.50
Italpollina x Trainer x Ecaterina 20.43 89.17 -2.48000
Orgevit x Trainer x Aurie de Bacău 21.79 95.11 -1.120
Phenix x Trainer x Aurie de Bacău 21.24 92.71 -1.6700
Italpollina x Trainer x Aurie de Bacău 20.29 88.56 -2.62000
Control (exp. average) 22.91 100.00 Mt
As far as trial results in Table 4 above are The second morphological feature analysed in
concerned. we need to note the very significant this trial was common bean pod width with
positive differences in the length of the pods direct impact on yield quality and quantity. We
(2.59-4.19 cm) in the Goldfield common bean made phonological observations and biometric
cultivar treated with the foliar fertiliser measurements on pod width. Trial results are
Cropmax and the three basic fertilisers. shown in Table 5 below.
Table 5. Influence of supplementary fertilisation on climbing common bean pod width

Supplementary fertilisation Pod width (cm) Relative values (%) Difference / Significance
Cropmax 2.05 102.20 0.04***
Lithovit 1.97 98.40 -0.03000
Trainer 1.99 99.40 -0.010
Comparative analysis of the unilateral influence Table 6 below shows trial results regarding the
of supplementary fertilisation during vegetation unilateral influence of basic (organic)
on climbing common bean pod width shows a fertilisation on climbing common bean pod
very significant positive difference (+0.04 cm) width.
when using the product Cropmax.
354
Table 6. Influence of basic fertilisation on climbing common bean pod width (Didactic Base Timişoara, 2016-2017)
Basic fertilisation Pod width (cm) Relative values (%) Difference/Significance
Orgevit 1.95 97.50 -0.05000
Phenix 2.04 101.90 0.04***
Italpollina 2.01 100.60 0.01**
As for the influence of basic fertilisation on (cultivar) is definitory in higher quality and
common bean pod width, there is a very quantity yields. We noted the unilateral
significant positive difference (+0.04 cm) when influence of the common bean cultivar used in
applying the fertiliser Phenix on a soil treated the trial on climbing common bean pod width
with more potassium. The genetic factor in table 7 (Mercati et al., 2013).
Table 7. Influence of cultivar on climbing common bean pod width (Didactic Base Timişoara, 2016-2017)
Cultivar Pod width (cm) Relative values (%) Difference / Significance
Goldfield 2.18 108.90 0.18***
Ecaterina 1.95 97.30 -0.05000
Aurie de Bacău 1.88 93.80 -0.12000
In this case, the Goldfield climbing common Trial data regarding the interdependence
bean cultivar is above the trial mean from the between fertiliser combinations and common
perspective of pod width (2.18 cm). bean cultivars are shown in Table 8 below.
Table 8. Interdependence of the combination basic fertilisation x supplementary fertilisation x cultivar
on climbing common bean pod width (Didactic Base Timişoara, 2016-2017)
Factor combination Pod width (cm) Relative values (%) Difference/Significance
Orgevit x Cropmax x Ecaterina 1.86 93.00 -0.14000
Phenix x Cropmax x Ecaterina 2.02 101.00 0.02*
Italpollina x Cropmax x Ecaterina 1.97 98.50 -0.0300
Orgevit x Cropmax x Aurie de Bacău 1.86 93.00 -0.14000
Orgevit x Lithovit x Goldfield 2.19 109.50 0.19***
Phenix x Lithovit x Goldfield 2.13 106.50 0.13***
Italpollina x Lithovit x Goldfield 2.12 106.00 0.12***
Orgevit x Lithovit x Ecaterina 1.78 89.00 -0.22000
Phenix x Lithovit x Ecaterina 2.03 101.50 0.03**
Orgevit x Trainer x Goldfield 2.18 109.00 0.18***
Orgevit x Trainer x Ecaterina 1.80 90.00 -0.20000
Phenix x Trainer x Ecaterina 2.04 102.00 0.04**
Control (exp. average) 2.00 100.00 Mt
355
Based on comparative assessment of the The quality of climbing common bean also
combinations of trial factors we noted again depends on pod diameter.
very significant positive differences in common From this perspective, we assessed the
bean pod width (0.19-0.29 cm) in the Goldfield unilateral influence of supplementary
common bean cultivar fertilised fertilisation on common bean pod diameter.
supplementarily with Cropmax with the three Trial results are shown in Table 9 below.
basic fertilisations.
Table 9. Influence of supplementary fertilisation on climbing common bean pod diameter
Supplementary fertilisation Pod diameter (cm) Relative values (%) Difference / Significance
Cropmax 0.691 101.61 0.01***
Lithovit 0.670 98.52 -0.01000
Trainer 0.677 99.55 -0.003000
Trial data shown in the table above show a very As in the morphological features analysed
significant positive difference in pod diameter above, we assessed the unilateral influence of
when using the foliar fertiliser Cropmax given basic fertilisation on common bean pod
its macro- and micro-element composition. diameter (Table 10).
Table 10. Influence of basic fertilisation on climbing common bean pod diameter
Basic fertilisation Pod diameter (cm) Relative values (%) Difference / Significance
Orgevit 0.671 98.82 -0.008000
Phenix 0.684 100.73 0.005***
Italpollina 0.683 100.58 0.004***
Comparing the three basic fertilisations of A synthesis of trial results regarding the
climbing common bean cultivated in solaria. unilateral influence of climbing common bean
we can note the very significant positive cultivar on pod diameter is shown in Table 11
differences when using Phenix and Italpollina. below.
Table 11. Influence of cultivar on climbing common bean pod diameter

Cultivar Pod diameter (cm) Relative values (%) Difference/Significance
Goldfield 0.717 105.50 0.03***
Ecaterina 0.699 102.90 0.01***
Aurie de Bacău 0.622 91.50 -0.06000
From the perspective of pod diameter, we noted Trial results regarding the interdependence
the very significant positive differences in the between treated soil and common bean cultivar
common bean cultivars Goldfield and on the diameter of common bean pods are
Ecaterina. shown in Table 12 below.
356
Table 12. Interdependence of the combination basic fertilisation x supplementary fertilisation x cultivar
on climbing common bean pod diameter (Didactic Base Timişoara, 2016-2017)
Factor combination Pod diameter (cm) Relative values (%) Difference/Significance
Orgevit x Cropmax x Ecaterina 0.729 106.57 0.045***
Phenix x Cropmax x Ecaterina 0.735 107.45 0.051***
Italpollina x Cropmax x Ecaterina 0.695 101.60 0.011*
Orgevit x Cropmax x Aurie de Bacău 0.628 91.81 -0.056000
Orgevit x Lithovit x Goldfield 0.683 99.85 -0.001
Phenix x Lithovit x Goldfield 0.697 101.90 0.013*
Italpollina x Lithovit x Goldfield 0.749 109.50 0.065***
Orgevit x Lithovit x Ecaterina 0.712 104.09 0.028***
Phenix x Lithovit x Ecaterina 0.717 104.82 0.033***
Orgevit x Trainer x Goldfield 0.695 101.60 0.011
Orgevit x Trainer x Ecaterina 0.716 104.67 0.032***
Phenix x Trainer x Ecaterina 0.712 104.09 0.028***
Control (average exp.) 0.684 100.00 Mt
As for the comparative analysis of the fertilisation with Cropmax on the three basic
combination of trial factors, we can say here fertilisations;
are very significant positive differences in pod - Climbing common bean pod diameter is very
diameter in the common bean cultivar significantly influenced by the three foliar
Goldfield cultivated on a soil treated with fertilisers (Cropmax, Trainer and Lithovit) on
Italpollina and treated with foliar fertilisers. the soil with basic fertilisation (Italpollina);
- From the perspective of climbing common
CONCLUSIONS bean pod diameter, we noted the Ecaterina
common bean cultivar with trial results close to
Based on trial results in the three climbing those of Goldfield common bean cultivar;
common bean cultivars cultivated in solaria - Ensuring optimum fertilisation rate with
with different basic and supplementary fertili- macro- and micro-elements during vegetation
sation, we can draw the following conclusions: in climbing common bean cultivated in solaria
- Comparative assessment of the three climbing influences morphological features and.
common bean cultivars from the perspective of Therefore, yielding capacity.
the three morphological features points out the
Goldfield common bean cultivar; REFERENCES
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358
THE EFFECTS OF DIFFERENT NITROGEN DOSES ON YIELD

AND NUTRIENT UPTAKE OF ROCKET (ERUCA SATİVA) PLANT
Volkan YORUK, Hasan DURUKAN, Handan SARAÇ, Ahmet DEMIRBAS,

Tolga KARAKOY
Cumhuriyet University, Vocational School of Sivas,

Department of Crop and Animal Production, Sivas, Turkey
Abstract
The present study was conducted to investigate the effects of different nitrogen doses on yield and nutrient uptake of
rocket plant. The study has been carried out with three repetitions according to the experimental pattern of randomized
plots in the plastic pots with the capacity of 3 kg under the greenhouse conditions. Nitrogen doses; 0 mg N kg-1, 50 mg
N kg-1, 100 mg N kg-1, 150 mg N kg-1, 200 mg N kg-1and were applied as CaNO3.4H2O form. The rocket plant was
harvested after 60 days of sowing and various traits like; shoot dry weight, macro- and micro-elements concentrations
were determined. The results of the study have shown that the highest shoot dry weight was determined in 200 mg N kg-
1
application with 4.45 g pot-1. In addition, the highest N concentration (5.80 % N) wa sobserved in 200 mg N kg-
1
application, P concentration in 50 mg N kg-1application (0.34% P), K concentration in 100 mg N kg-1 application
(7.01% K), Mg and Ca concentrations in 150 mg N kg-1 application (0.92% Mg, 1.35% Ca) were determined.
Considering all treatments together, it was observed that increasing nitrogen doses increased yieldand macro and
microelements concentrations of rocket plant.
Key words: rocket, nitrogen, yield, nutrient uptake.
INTRODUCTION (N) is one of these nutritional materials and it is

necessary for the formation of new cells in the
Rocket (Eruca sativa) is an endemic species of plants. The growth rate of the plants declines
Brassicaceae family and a single year due to the nitrogen deficiency. Particularly the
herbaceous plant. This species is widely vegetative development of the plants is
cultivated such Mediterranean countries as negatively affected.
Italy, Greece and Turkey in particular (Aegean, Leave and root systems of the plants is
Marmara and Mediterranean regions) (Morales relatively weakened. Similarly, development of
and Janick, 2002; Barlas et al., 2011; Baser, roots and particularly the branching of the roots
2016). Fresh leaves of rocket plant have long is weakened. Flowering and fruit production
been used in salads of Mediterranean cuisine rates decrease and fruits are relatively small in
(Başer, 2016). With the current increase in the size (Foth, 1984; Aktaş and Ateş, 1998;
consumption of green vegetables which are Kantarcı, 2000; Boşgelmez et al., 2001; Güzel
beneficial to human health, the economic et al., 2004; Fageria, 2009; Kacar and Katkat,
potential rocket plant has steadily increased 2010; Bolat and Kara, 2017).
recently (Eşiyok et al., 2006). Although nitrogen is the main nutritient
In order for being able to harvest quality yield element, which is the most absorbed nutrient in
in plentitude in vegetable cultivation (Stewart comparison to other elements, it is one of the
et al., 2005), plant nutrients are emphasized in most deficient nutrient elements. A great part
recent years (Adediran et al., 2004; Naeem et of soils on the Earth is subject to nitrogen
al., 2006). Plants need at least 17 plant deficiency.
nutritional materials or elements to achieve and The main source of nitrogen in the soil is
demonstrate optimal growth and development organic materials. Proteins, amino acids,
(White, 2006; Gardiner and Miller, 2008; nucleic acids, enzymes, chlorophylls, ATP,
Fageria, 2009; Bolat and Kara, 2017). Nitrogen ADP are among significant organic materials
359
including nitrogen. As a result of the gradual department. Soils were sieved through 2 mm
decomposition of the organic materials, plants sieve and 3 kg air-dried soils were placed in
can make use of the nitrogen contained in these experimental pots.
decomposed materials. Some physical and chemical characteristics of
The soils of Turkey, which are particularly soil are provided in Table 1. Five levels of
deficient in organic materials, are relatively nitrogen; 0 mg N kg-1, 50 mg N kg-1, 100 mg N
poor in terms of nitrogen (Çepel, 1996; Aktaş kg-1, 150 mg N kg-1 and 200 mg N kg-1 (in
and Ateş, 1998; Boşgelmez et al., 2001; Güzel CaNO3.4H2O form) were applied. Before
et al., 2004; Gardiner and Miller, 2008; sowing, 100 mg kg-1 P and 125 mg kg-1 K (in
McCauley et al., 2009; Bolat and Kara, 2017). the form of KH2PO4), 2.5 mg kg-1Zn (in the
For this reason, chemical fertilizers which are form of ZnSO4.7H2O) and 2.5 mg kg-1 Fe (in
commonly used for preventing nutritional the form of Fe-EDTA) were applied to each pot
imbalance in plants and satisfying needs of as basic fertilizers.
plants for nutritional elements are mainly
composed of nitrogen content (Kacar and Plant analyses
Katkat, 2010). Nitrogen has a significant effect Leaf samples were taken from the rocket plants
on the plant quality. 45 days after the sowing. Vegetative parts of
The conducted research has demonstrated a the plants were washed through tap water,
direct correlation between nitrogen levels and rinsed respectively through distilled water, 0.1
quality of the plant yield. In accordance with N HCl solution and twice though again distilled
the increase in the nitrogen dose applied to the water. They were placed over coarse filter
plants, the N concentration in the plant grain papers and excess water over them was
and accordingly the grain quality also removed.
demonstrate increases (Marschner, 1997). Plant parts were then placed in separate paper
However, in recent years, it has been detected bags and dried at 65C until a constant weight.
that the remaining nitrogen in the soil from the Following the measurement of dry weights, dry
high concentration nitrogen fertilizer samples were ground in a plant mill. About 0.2
applications results in environmental pollution g of ground samples were wet digested in
(Zand-Parsa et al., 2006; Gollany et al., 2004; H2O2-HNO3 acid mixture in a microwave oven.
Beman et al., 2005) and leads to accumulation Resultant slurry was then completed to 20 ml
of materials in plants detrimental to human with distilled water and filtered through blue-
health (Ruiz and Romero, 1999). band filter paper. Samples were then subjected
For this reason, the management and program- to P colorimetric K, Ca, Mg, Zn, Mn, Fe and
ming of nitrogen in vegetable cultivation are of Cu readings at 882 nm (Murphy and Riley,
due significance (Adiloğlu et al., 2015). 1962) in an AAS (Atomic Absorption
The aim of the present study is determining the Spectrophotometer) (Shimadzu AA-7000)
effects of administering nitrogen in varying (Kaçar and İnal, 2008). N contents were
quantities, which is a relatively significant determined with Kjeldahl distillation method
nutrient element, on the productivity of and (Bremner, 1965).
absorption of nutrient elements by rocket plant.
Data assessment
MATERIALS AND METHODS Experimental results were subjected to variance
analyses (ANOVA) separately in accordance
This study was conducted at greenhouses of with randomized plots experimental design.
Plant and Animal Production Department of SPSS 22.0 Windows software was used for
Cumhuriyet University Sivas Vocational statistical analyses. Means were compared with
Collage. Experiment was conducted in Tukey’s test at P<0.05. Correlation analysis
randomized plots design with 3 repetitions. was performed to assess the relationships
Experimental soils were taken from 0-30 cm between the treatments.
soil profile of experimental fields of the
360
Table 1. Some physical and chemical properties of soil
Soil Property Depth
(0-30cm)
pH 7.26
Lime (%) 17.0
Salt (%) 0.034
Organic matter (%) 1.38
Texture SiCL
Total N (%) 0.087
Available P (kg ha-1) 47.3
Available K (kg ha-1) 744.8
Available Fe (mg kg-1) 3.11
Available Mn (mg kg-1) 1.04
-1
Available Zn (mg kg ) 0.41
Available Cu (mg kg-1) 1.22
RESULTS AND DISCUSSIONS investigated and results are presented in Figure

1. The greatest dry matter production (4.45 g
Effects of different nitrogen treatments on dry pot-1) was obtained from 200 mg N kg-1
matter production of rocket plant were treatment (Figure 1).
Figure 1. Effects of different nitrogen fertilizer on shoot dry matter of rocket plant
This treatment was followed by 100 mg N kg-1 and made harvests in June, July and August. It
and 150 mg N kg-1 treatments respectively with was reported that marketable product quantity
3.86 g pot-1 and 3.75 g pot-1 dry matter increased from 4.4 ton ha-1 to 8.3 ton ha-1
productions which were placed in the same (about doubled) with N treatments. Eşiyok et
statistical group. The lowest dry matter al. (1998) investigated the effects of different
production was observed in control treatment nitrogenous fertilizers on yield and nutrient
(0 mg N kg-1) with 0.61 g pot-1. Increasing dry uptake of rocket plants. Yield and nutrient
matter production of rocket plants were uptakes varied with nitrogen doses and the
observed with increasing N doses. Haag and greatest yield was obtained from 160 kg ha-
1
Minami (1988) applied four different nitrogen nitrogen treatment. Researchers also reported
doses (0, 100, 200, 300 kg ha-1) to rocket plants decreasing leaf nitrogen, phosphorus,
361
potassium and other nutrient contents with the (110, 175 and 240 mg l-1) to the plants in 4
aging of the leaves as compared to young replications and indicated improved plant
leaves harvested at early periods. Demiral et al. growth with the experimental treatments.
(2009) investigated the effects of N treatments Hanaka et al. (2016) applied two different
on plant growth and nitrate (NO3-) nitrate (0.3, 0.6 g N dm-3 of medium) and 3
accumulation of four different winter different potassium (0.3, 0.6, 0.9 g K dm-3 of
vegetables including chard (Beta vulgaris cv. medium) doses to rocket plants and reported
Chard), rocket (Eruca sativa), lettuce (Lactuca dry matter productions under experimental
sativa) and spinach (Spinacia oleracea). treatments as between 9.2-10.1 g.
Researchers applied three different N doses
Table 2. Effects of different nitrogen fertilizer on N, P and K concentrations of rocket plant (%)
N P K
Treatments
(%)
0 mg N kg-1 1.91 ±0.02d 0.15 ±0.01d 3.09 ±0.05e
50 mg N kg-1 2.75 ±0.25c 0.34 ±0.02 a 4.40 ±0.02 d
100 mg N kg-1 4.97 ±0.18 b 0.28 ±0.01 b 7.01 ±0.11 a
150 mg N kg-1 5.16 ±0.04 b 0.28 ±0.03 b 6.71 ±0.04 b
200 mg N kg-1 5.80 ±0.06 a 0.22 ±0.00 c 5.51 ±0.29c
P<0.05
Considering the effects of different nitrogen The greatest K concentration (7.01% K) was
treatments on N concentration of the rocket obtained from 100 mg N kg-1 treatment. K
plants, the greatest value (5.80% N) was concentrations decreased after this dose. All
obtained from the greatest nitrogen dose of 200 nitrogen doses significantly increased % N, %
mg N kg-1 treatment. P and % K concentrations of rocket plants as
Plant N concentration of the control (0 mg N compared to the control treatment. Nurzyńska-
kg-1) treatment was 1.91% N. As it was in dry Wierdak (2015) applied different nitrogen and
matter productions, increasing plant N potassium doses to rocket plants and reported
concentrations were observed with increasing % P concentrations of the plants as between
nitrogen doses. Similarly, Hanaka et al. (2016) 0.51-0.61% P and % K concentrations as
also reported increasing leaf N concentrations between 5.30-6.76% K. Barlas et al. (2011) in a
of rocket plants with increasing N doses survey study, collected rocket plants from 30
(especially when combined with S). different fields and reported %N concentrations
The greatest P concentration (0.34% P) was as between 2.94-5.23% N, % P concentrations
observed in 50 mg N kg-1 treatment and plant P as between 0.12-0.27% P and K concentrations
concentrations decreased after this N dose. as between 3.99-5.98% K.
Table 3. Effects of different nitrogen fertilizer on Ca and Mg concentrations of rocket plant (%)
Ca Mg
Treatments
(%)
0 mg N kg-1 0.54 ±0.02 e 0.63 ±0.03 bc
50 mg N kg-1 1.08 ±0.01 c 0.88 ±0.07 ab
100 mg N kg-1 1.22 ±0.03 b 0.90 ±0.03 a
150 mg N kg-1 1.35 ±0.02 a 0.92 ±0.03 a
200 mg N kg-1 0.93 ±0.03 d 0.59 ±0.03 c
P<0.05
Considering the Ca concentrations of the rocket treatment and the greatest value (1.35% Ca)
plants, the lowest value (0.54% Ca) was was obtained from 150 mg N kg-1 treatments
obtained from the control (0 mg N kg-1) (Table 3). Barlas et al. (2011) reported leaf Ca
362
concentrations of rocket plants as between mg N kg-1 treatment (0.90% Ca) which was
2.20-3.55% Ca. Bukhsh et al. (2007) reported placed in the same statistical group. In previous
the Ca concentration as 1900 µg g-1 Ca in seeds studies, Mg concentrations of rocket plants
of rocket plants and as 700 µg g-1. Ca in the were reported as between 37-41 mg 100 g-1
leaves of rocket plants. Similar to Ca (Haag and Manami, 1998) and as 46 mg 100 g-1
concentrations, the greatest Mg concentration (Bianco, 1995). Eşiyok et al. (2006) reported
(0.92% Mg) was obtained from 150 mg N kg-1 Mg content of organic fertilizer-treated rocket
treatment. This treatments was followed by 100 plants as between 0.19-0.23% Mg.
Table 4. Effects of different nitrogen fertilizer on Fe, Zn, Mn
and Cu concentrations of rocket plant
Fe Zn Mn Cu
Treatments
(mg kg -1)
0 mg N kg-1 73.4 ±1.31 d 10.9 ±0.48 c 26.8 ±1.05 d 4.4 ±0.04 c
50 mg N kg-1 163.4 ±14.07 a 30.2 ±2.86 a 27.7 ±0.53 cd 5.5 ±0.32 b
100 mg N kg-1 133.6 ±12.52 b 24.6 ±0.01 b 29.4 ±0.23 c 5.9 ±0.08 b
150 mg N kg-1 151.3 ±5.23 a 24.6 ±0.10 b 35.7 ±0.21 b 6.3 ±0.16 ab
200 mg N kg-1 97.6 ±1.39 c 12.1 ±0.07 c 42.9 ±2.18 a 7.1 ±0.71 a
P<0.05
Considering the micro element concentrations and Cu concentrations were obtained from 200
of rocket plants (Table 4), the greatest Fe mg N kg-1 treatment. While the greatest Mn
concentration was obtained from 50 mg N kg-1 concentration was 42.9 mg kg-1Mn, the greatest
treatment (163.4 mg kg-1). This treatment was Cu concentration was observed as 7.1 mg kg-1
followed by 150 mg N kg-1 (151.3 mg kg-1 Fe). Cu. Bukhsh et al. (2007) reported Fe
As compared to control treatment, all nitrogen concentration of rocket plants as 60.62 µg g-1
doses increased plant Fe concentrations. Zn Fe, Zn concentration as 56.1 µg g-1 Zn, Mn
concentrations of the rocket plants also concentration as 19.0 µg g-1Mn and Cu
increased with increasing nitrogen doses. concentration as 32.0 µg g-1 Cu; reported leaf
The lowest Zn concentration (10.9 mg kg-1 Zn) Fe concentration as 37.5 µg g-1 Fe, leaf Zn
was obtained from 0 mg N kg-1 control concentration as 1.12 µg g-1 Zn, leaf Mn
treatment and the greatest Zn concentration concentration as 10.6 µg g-1Mn and leaf Cu
(30.2 mg kg-1 Zn) was obtained from 50 mg N concentration as 21.0 µg g-1 Cu. In a survey
kg-1 treatment. As it was in P concentrations, study, Fe concentration of rocket plants was
Zn concentrations of rocket plants decreased reported as 350.78 mg kg-1 Fe, Zn concen-
after 50 mg N kg-1 treatments. Increasing tration as 64.86 mg kg-1 Zn, Mn concentration
nitrogen doses also increased Mn and Cu as 40.58 mg kg-1Mn and Cu concentration as
concentrations of the plant. The greatest Mn 5.37 mg kg-1 (Barlas et al., 2011).
Table 5. Correlation values of parameters evaluated in the study
Parameters SDW*** N P K Mg Ca Fe Zn Mn Cu
SDW 1
N .907** 1
P .536* .194 1
K .818** .850** .468 1
Mg .295 .197 .613* .631* 1
Ca .767** .640* .780** .882** .781** 1
Fe .498 .188 .949** .508 .768** .834** 1
Zn .335 .029 .948** .421 .730** .755** .948** 1
Mn .545* .398 .138 -.015 -.468 .004 .007 -.148 1
Cu .913** .891** .293 .678** .113 .586* .282 .077 .661** 1
*Significant at P<0.05 **Significant at P<0.01 ***SDW=Shoot Dry Weight
363
Considering the correlations among investi- Barlas N.T., Irget M.E., Tepecik M., 2011. Mineral
gated parameters, it was observed that SDW content of the rocket plant (Eruca sativa). Afr. J.
Biotechnol., 10 (64): 14080-14082.
had significant positive correlations with N, K, Başer K.H.C., 2016. Roka (Eruca sativa Mill.). Bağ-
Ca and Cu (P<0.01); there was significant Bahçe (68) 26-27.
positive correlation between P and Mn Beman J.M., Arrigo K., Matson P.M., 2005. Agricultural
(P<0.05) (Table 5). There was significant runoff fuels large phytoplankton blooms in
positive correlations also between N and K, vulnerable areas of the ocean, Nature, 434: 211-214.
Bianco V.V., 1995. Rocket an ancien tunder utilized
between Ca and Cu; P had positive correlations vegetable crop and its potential, Rocket genetic
with Mg, Ca, Fe and Zn; there were positive resources network. Repot of the first meeting, 13-15
correlations between K and Mg, between Ca November, Libson, Resources Institue, Rome, Italy,
and Cu; and Mg had significant positive pp: 35-37.
correlations with Ca, Fe and Zn (P<0.01); Ca Bolat İ., Kara Ö., 2017. Plant Nutrient Elements:
Sources, Functions, Toxicity and Disorders. Journal
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were positive correlations between Fe and Zn Turkish).
and between Mn and Cu. Boşgelmez A., Boşgelmez İ.İ., Savaşçı S., Paslı N.,
2001. Ecology - II (Soil), Baskent Klise Press.,
CONCLUSIONS Kızılay-Ankara/Turkey (in Turkish).
Bremner J.M., 1965. Total nitrogen. Methods of soil
analysis. Part 2. Chemical and microbio-logical
The present study was conducted to investigate properties. (methods of soil anb), 1149-1178.
the effects of different nitrogen doses on yield Bukhsh E., Malik S.A., Ahmad S.S., 2007. Estimation of
and nutrient uptake of the rocket plants. nutritional value and trace elements content of
Present findings revealed increasing dry matter Carthamus oxyacantha, Eruca sativa and Plantago
ovata. Pak. J. Bot., 39 (4): 1181-1187.
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the other hand, 150 mg N kg-1 treatments were some leafy vegetables. ADÜ Ziraat Fakültesi Dergisi,
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366
THE IMPACT OF DIFFERENT METHODS USED

FOR IMPROVING FLOWER BIND UPON THE YIELD
OF SOME TOMATO HYBRIDS CULTIVATED ON MINERAL WOOL
SUBSTRATE IN INDUSTRIAL GREENHOUSES
Alexandra BECHERESCU1, Gheorghiţa HOZA2, Maria DINU3,

Olimpia IORDĂNESCU1, Ioan SĂRAC1, Daniel POPA1
1
from Timisoara, 119, Calea Aradului, Timișoara, Romania
2
3
University of Craiova, 13 A.I. Cuza Street, Craiova, Romania
Abstract
Obtaining high and superior qualitative tomato yields, in conditions of forced culture system, being economically
profitable, depends on modernizing the culture technology. In classical Venlo industrial greenhouses, modernization is
possible till the technical limit allowed by their construction type. Among the technological links that can be
modernized, and to which we refer in this study, are the following: cultivation of performant hybrids with high
quantitative and qualitative potential (Noralee F1 and Marissa F1); fertilization during vegetation with modern
chemicals, completely soluble (Yara and Haifa Chemicals), applied using drip irrigation system; using mineral wool as
culture substrate (Grodan) and applying new and efficient methods for improving flower bind (pollination with
bumblebees - Biobest or Natupol). This article presents the impact of all these modernized technological links upon the
quantitative and qualitative tomatoes yields. Tomato-Stim determined large tomato yields, but the best results upon the
yield quantity and quality were obtained from those tomato plants where natural pollination was improved by the use of
bumblebees. Most of the results showed that the quality of the obtained yields (namely Extra+Ist quality) finds itself in a
reversed proportionality with the production quantity per hectare. The influence of both graduation of the fertigation
systems are substantiated in the very close quantitative production and from a statistic point of view were not covered,
as there were no significant production differences. Results lead us to recommend the cultivation of Noralee F1 hybrid,
the use of any of the fertilizers Yara and Haifa Chemicals via fertigation and the use of bumblebees (Biobest or
Natupol) to improve the tomato flower binding and to obtain superior quality fruits.
Key words: bumblebees fertigation, quality.
INTRODUCTION represents an important technological link in

the modernization process as it determines the
Modernizing the culture technology for improvement of other technological links,
vegetable cultivated in a protected or forced which are irrigation and fertilization at the
system, consists in improving some techno- same time by using the drip-irrigation system
logical links (improving flower bind degree, (fertigation). (Horgoş A. et al., 2005).
using modern fertilizers, performant hybrids Bumblebees are important pollinators for
etc.) so that the effect would be and efficient tomato crops. Foragers are capable of assessing
yield, considering its productivity, quality and the pollen reward of the flowers, the first
economic efficiency (Horgoş A. et al., 2002). flower visit is the most effective concerning
Today, tomatoes is widely cultivated pollen removal and, consequently, pollination,
throughout the world and adapted into many and small foragers are as efficient in pollinating
cultivars (Olmstead and Palmer, 1997). tomato flowers as are big ones. In line with
The replacement of soil, as natural culture some earlier studies (Buchmann and Cane
substrate, in greenhouses with an artificial 1989; Harder, 1990; Shelly et al., 2000)
culture substrate, in this case mineral wool, provide clear evidence that bees are indeed
367
capable of perceiving the amount of pollen The culture was established in spring, the
obtained while visiting a flower (Nunes-Silva beginning of March (the 2nd - 3rd of March
Patricia et al., 2013). 2015), when the seedling had 65 days and it
The cultivated plant is autogamous. However, was planted to density of 24,288 plants/ha. The
one of the features of the genus is the poricidal experiment is considered to be poly-factorial,
opening of its anthers, which requires the having the following factors:
agitation of the flowers by wind and/or the Factor A - Hybrids with undetermined growth
presence of pollinators that vibrate their cultivated on mineral wool: a1 - Noralee F1; a2 -
indirect flight muscles for the release of pollen Marissa F1.
grains, even in cultivated varieties of tomatoes Factor B - Fertigation system: b1 - Fertigation
and especially in the still air of greenhouses with completely soluble chemicals: Yara (Yara
(Kevan et al., 1991; Morandin et al., 2001b). Mila Cropcare; Yara Mila Ferticare I-III; Yara
Teppner (2005), while conducting studies on Mila Folicare B; Folicare Zn; Folicare Cu;
tomato plants in central Europe, observed that KNO3 Krista; Ca (NO3)2 , Calcinit; K3PO4);b2 -
bees, such as Bombus and Lasioglossum, can Fertigation with completely soluble chemicals:
be good pollinators of the flowers by vibrating Haifa-Chemicals (Multicote 4, Multicote 4 with
their anthers easily. microelements - ME, magnesium nitrate,
Pollination is an important component of crop complex soluble fertilizers N-P-K, mono-
production for many cultivated plants. Through potassium phosphate - MPK).
pollination research focused on crop plants, Factor C - Improvement of flower bind (fecun-
agricultural practices become better designed to dation): c1 - Control, no stimulants, natural
efficiently produce high quality crops (Morse pollination; c2 - Sprayings with Tomato-Stim
A., 2009). stimulant (artificial stimulation); c3 - Mecha-
In conclusion, the combined influence of nical methods (shaking plants, minting the
technological works listed above, in correlation plant supporting wires, producing air streams
with the micro-climate factors from industrial by using the pulverizer); c4 - Using bumblebees
heated greenhouses; assure high yields, (Biobest, Natupol) to improve natural
qualitatively superior, which determine the pollination.
profitability of tomato cultures. The culture technology was distinctive by three
modern works: the use of artificial culture
MATERIALS AND METHODS substrate (mineral wool used as “matress”), the
drip irrigation system (Netafim-Israel instal-
Researches on the impact of different methods lation type) and the fertilization done with
for improving flower binding in tomato hybrids completely soluble chemicals using the drip
culture with undetermined growth, on mineral irrigation installation (fertigation process).
wool, were developed in classic Venlo indus- The researches were done in order to determine
trial greenhouses from Agro-Codlea Company the efficacy of tomato culture when cultivating
(placed in the Western part of Romania, in hybrids with undetermined growth, with high
Arad County). The experiment was developed productive potential, cultivated on artificial
upon two Dutch hybrids with undetermined substrate in conditions of modern technology,
growth: Noralee F1 (generative hybrid) new which is still limited by the Velno classical
hybrid belonging to Enza Zaden company and greenhouse.
Marissa F1, belonging to Royal Sluis company,
which is vigorous and high performances RESULTS AND DISCUSSIONS
regarding fructification and fruits’ quality.
The hybrids were observed regarding their The experimental results presented in tables 1
productive and qualitative potential under the and 2 and figure 1 are the expression of the
impact of two fertigation systems (with two interactions between the three experimental
completely soluble modern chemical fertilizers- factors, materialized in the different production
Yara and Haifa Chemicals) and also by using and quality levels, related to the variability of
different methods for improving flower bind production elements (average number of fruits
(fecundation). per plant and average weight of one tomato
fruit). The number of fruits per plant is clearly
368
influenced by the method used for improving 139.8 t/ha in a1b2c4 (Noralee F1-Haifa
flower binding (factor C), the values being of Chemicals - Bumblebee pollination).
118.4% (c3 - Mechanical methods) and 140.6% Considering its qualitative production, the extra
(c2 - Sprayings with Tomato-Stim stimulant) and first quality yield shares of the total yield
compared to the control c1 - No stimulants, are relatively close, being 82.0% in a1 - Noralee
natural pollination (100%). The same rule F1 and 80.2% in a2 - Marissa F1.
applies to the average weight of fruits, the Variations percentage share of extra and first
maximum value being obtained under the quality production have values between 84.4%
impact of c3 (Mechanical methods - plants’ in a1b1 (Noralee F1-Yara), 79.5% in a1b2
shaking, minting the plant supporting wires, (Noralee F1-Haifa Chemicals) and 82.4% in
producing air streams with the pulverizer) and a2b1 (Marissa F1-Yara), 78.0% in a2b2 (Marissa
c4 (Using bumblebees for natural pollination), F1-Haifa Chemicals).
compared to the other two methods, c1 (No Preceding with the analysis of the summary the
stimulants, natural pollination) and c2 3rd table regarding the experimental result we
(Sprayings with Tomato-Stim stimulant). arrive to the following:
The hierarchy levels of production are in direct -In all the graduations of factor C (method of
proportionality rule with the number of fruits flower binding improvement), the production
per plant and the average weight of a fruit. achieved under the influence of b1-Yara is on
Production quality levels, in terms of its an average higher than under the influence of
quantitative proportionality, do not follow the b2-Haifa Chemicals, the difference being rather
same route under the direct influence of all very small (1.6 t/ha, namely 1.3%); the highest
graduations of factor C (Method of improving difference being registered in C4 (the natural
the binding of flowers). There is a deviation pollination via bumblebees), these being 5.4
under the impact of c2 (Sprayings with Tomato- t/ha, namely 4.0% (c4b1a1 = 145.2
Stim stimulant - artificial stimulation), where at t/ha→106.9% and c4b2a1 = 139.8
a production point almost equal to the one t/ha→102.9%);
obtained under the impact of c4 (Using -Compared to MX1 [ܿହ ൌ ሺܿଵ ൅ ‫ ڮ‬൅ ܿସ ሻȀͶሿ =
bumblebees for natural pollination) the 120.0 t/ha→100.0%), the production from b1
difference is not significant (-1.5 t/ha), while (Yara) is 120.8 t/ha→100.7%, and for b2 (Haifa
the extra and first quality production represent Chemicals) is 119.2 t/ha→99.3%, a difference
72.5% of the total, which is with 13.8% less of 1.6 t/ha→1.4% being very low; the
than under the impact of c4 (Using bumblebees difference is similar also in the case of MX2
for natural pollination), where the share is ሾܿ଺ ൌ ሺܿହ െ ܿଵ ሻȀ͵ሿ →100.0%, meaning 1.7
86.3%. t/ha →1.3%;
Under the impact of factor B’s graduations - The highest productions are obtained under
(fertigation system), the production elements the influence of graduation c2 (Tomato-Stim),
and the obtained quantities are almost equal 134.3 t/ha → 153.3% compared to c1 – Mt and
(the differences varying between 0.8-2.8 t/ha, under c4 (bumblebees pollination), 135.8 t/ha
respectively 0.6-2.0%). Considering → 155.0% compared to c1–Mt;
production’s quality (extra and first quality), - The production quality of the tomatoes under
the difference between b1 (Yara) and b2 (Haifa the influence of c2 (Tomato-Stim) is lower
Chemicals) is significant, in the limits of 2.8% though, percentage wise down to 72.5% (97.4
(c1) and 5.4% (c2). t/ha E+Ist quality) from a total production of
Factor A (the hybrid) contributes to the 134.3 t/ha, compared to the best quality of the
differentiation of production levels, at the same same, namely the one from c4 (bumblebees
time quantitative and qualitative), in interaction pollination), of 86.3% (117.2 t/ha E+Ist quality)
with both factor B and C and their graduations. of a total of 135.8 t/ha;
Hybrid Noralee F1 has a medium production - The production quality obtained (Extra+Ist
level of 124.9 t/ha and a maximum of 145.2 quality) finds itself in a reversed
t/ha in a1b1c4 (Noralee F1-Yara-Bumblebee proportionality with the production quantity per
pollination), being followed by 140.5 t/ha in hectare;
a1b1c2 (Noralee F1-Yara-Tomato-Stim) and
369
- Out of the four methods of flower binding, (c4), where the average production per hectare
from a quantity point of view as well as quality as well as the quality had maximum worth
of tomato production, the method of using compared to the other graduations.
Biobest or Natupol bumblebees takes first place
Table 1. Experimental results on cultivating undetermined growth tomato hybrids
in industrial greenhouses on mineral wool substrate, Ist cycle - 2015
Average production
Average no.
of Average of which extra and
Factor C Factor B
Factor A fruits/plant weight/ % Ist quality
(Improvement of (Fertigation
(The hybrid) fruit kg/plant t/ha than production
flower bind) system)
piece/ (g/piece) c1
% t/ha %
pl
a1 - Noralee F1 32.6 100.0 114.7 3.738 90.8 103.7 76.0 83,7
b1-Yara a2 - Marissa F1 31.5 100.0 111.9 3.524 85.6 97.7 70.0 81,8
c1- Control, no Average a for c xb
1 1 32.1 100.0 113.3 3.631 88.2 100.7 73.0 82,8
stimulants, a1 - Noralee F1 32.5 100.0 114.3 3.714 90.2 103.0 73.2 81,2
natural b2-Haifa
a2 - Marissa F1 31.2 100.0 110.6 3.450 83.8 95.7 67.1 80,1
pollination chemicals
Average a for c1xb2 31.9 100.0 112.5 3.582 87.0 99.3 70.2 80,7
Average value of factor B for
32,0 100.0 112.9 3.607 87.6 100.0 71.6 81.7
c1
a1 - Noralee F1 46.9 143.9 123.3 5.785 140.5 104.6 106.9 76,1
b1-Yara a2 - Marissa F1 43.7 138.7 121.4 5.307 128.9 95.8 95.6 74,2
c2- Sprayings Average a for c2xb1 45.3 141.1 122.4 5.546 134.7 100.3 101.3 75,2
with Tomato-
a1 - Noralee F1 45.9 141.2 122.7 5.632 136.8 101.9 97.3 70,9
Stim stimulant b2-Haifa
(artificial a2 - Marissa F1 43.5 139.4 124.0 5.394 131.0 97.5 90.0 68,7
chemicals
stimulation) Average a for c2xb2 44.7 140.1 123.4 5.513 133.9 99.7 93.5 69,8
45,0 140.6 122.9 5.529 134.3 100.0 97.4 72.5
c2
a1 - Noralee F1 39.8 122.0 139.1 5.536 129.6 106.1 114.2 88,1
c3- Mechanical b1-Yara a2 - Marissa F1 37.0 117.5 129.3 4.784 116.2 95.2 100.2 86,2
methods (shaking Average a for c3xb1 38.4 119.6 134.2 5.060 122.9 100.7 107.2 87,2
plants, minting
a1 - Noralee F1 38.2 117.5 136.5 5.216 126.7 103.8 104.3 82,3
the plant b2-Haifa
supporting wires, chemicals a2 - Marissa F1 36.4 116.7 131.1 4.772 115.9 94.9 94.3 81,4
producing air Average a for c3xb2 37.3 116.9 133.8 4.994 121.3 99.3 99.3 81,9
streams) Average value of factor B for
37,9 118.4 134.0 5.027 122.1 100.0 103.3 84.6
c3
a1 - Noralee F1 42.9 131.6 139.3 5.978 145.2 106.9 129.9 89,5
b1-Yara a2 - Marissa F1 40.4 128.3 131.7 5.319 129.2 95.1 113.4 87,8
c4- Natural Average a for c xb
4 1 41.7 129.9 135.5 5.649 137.2 101.0 121.7 88,7
pollination using a1 - Noralee F1 41.7 128.3 138.0 5.756 139.8 102.9 118.0 84,4
bumblebees b2-Haifa
a2 - Marissa F1 40.6 130.1 130.8 5.311 129.0 95.0 107.1 83,0
(Biobest) chemicals
Average a for c4xb2 41.2 129.1 134.4 5.533 134.4 99.0 112.6 83,8
41,5 129.7 135.0 5.591 135.8 100.0 117.2 86.3
c4
Culture density: 24,288 plants/ha
370
Table 2. Experimental results on cultivating undetermined growth tomato hybrids
in industrial greenhouses on mineral wool substrate, Ist cycle - 2015
Average production
Average no. Average of which extra and
Factor A Factor B Factor C of fruits/plant weight/
% Ist quality
(The (Fertigation (Improvement of flower production
system) fruit kg/plant t/ha than
hybrid) bind)
piece/ (g/piece) c1
% t/ha %
pl
c1 - Control - natural
32.6 81.5 114.7 3.738 90.8 100.0 76.0 83,7
pollination
c2 - Stimulation with Tomato-
46.9 117.3 123.3 5.785 140.5 154.7 106.9 76,1
Stim
b1-Yara
c3 - Mechanical pollination 39.8 99.5 139.1 5.536 129.6 142.7 114.2 88,1
c4 - Bumblebee pollination -
42.9 107.3 139.3 5.978 145.2 159.9 129.9 89,5
Biobest
Average of factor C for a1xb1 40.5 101.3 129.1 5.209 126.5 139.3 106.8 84,4
a1 -
Noralee F1 32.5 81.3 114.3 3.714 90.2 100.0 73.2 81,2
pollination
45.9 114.8 122.7 5.632 136.8 151.7 97.0 70,9
Stim
b2-Haifa
chemicals c3 - Mechanical pollination 38.2 95.5 136.5 5.216 126.7 140.5 104.3 82,3
41.7 104.3 138.0 5.756 139.8 155.0 118.0 84,4
Biobest
Average of factor B for a1 40.0 100.0 128.5 5.144 124.9 * 102.5 82.0
31.5 82.7 111.9 3.524 85.6 100.0 70.0 81,8
pollination
43.7 114.7 121.4 5.307 128.9 150.6 95.6 74,2
Stim
b1-Yara
c3 - Mechanical pollination 37.0 97.1 129.3 4.784 116.2 135.7 100.2 86,2
40.4 106.0 131.7 5.319 129.2 150.9 113.4 87,8
Biobest
a2 -
Marissa F1 31.2 81.9 110.6 3.450 83.8 100.0 67.1 80,1
pollination
43.5 114.2 124.0 5.394 131.0 156.3 90.0 68,7
Stim
b2-Haifa
chemicals c3 - Mechanical pollination 36.4 95.5 131.1 4.772 115.9 138.3 94.3 81,4
40.6 106.6 130.8 5.311 129.0 153.9 107.1 83,0
Biobest
Average of factor B for a2 38.1 100.0 123.9 4.733 114.9 * 92.2 80.2
Culture density: 24,288 plants/ha.
371
Average no.of fruits/plant (pcs) Average weight of fruit (g) Average yield (t/ha) Average yield (kg/pl)
180,0 55,978
978 6,30
5,785 5,632 5,756
5,536 5,394
145,2
5,216 5,307 5,319 5,311
140,5
160,0 5,60
139,3
138,0
136,8
139,8
131,7
131,0
130,8
128,9
4,784 4,772
139,1
129,2
129,6
124,0
129,0
123,3
122,7
121,4
140,0 4,90
136,5
116,2
115,9
126,7
114,7
114,3
111,9
110,6
131,1
129,3
120,0 4,20
3,738
90,8
90,2
85,6
100,0 3,50
83,8
3,714 3,524 3,450
80,0 2,80
46,9
45,9
60,0 2,10
43,7
43,5
42,9
41,7
40,6
40,4
39,8
38,2
37,0
36,4
32,6
32,5
31,5
31,2
40,0 1,40
20,0 0,70
0,0 0,00
c4 – Bumblebees natural pollination


c3 – Mechanical methods used for

c1 – Control, no stimulents, natural


c2 – Sprayings w ith Tomato-Stim

pollination
pollination
pollination
pollination
pollination
pollination
pollination
pollination
b1-Yara b2-Haifa Chemicals b1-Yara b2-Haifa Chemicals
a1 - Noralee F1 a2 - Marissa F1
Figure 1. Experimental results on cultivating undetermined growth tomato hybrids in industrial greenhouses on mineral
wool substrate, Ist cycle - 2015
Table 3. Synthesis of experimental results on the non-determined growth tomato hybrids cultivated
in warm greenhouses with modernized technological works
Factor
Average production for:
experimental
Factor A Factor B Factor C
E+Ist % c1- E+Ist quality E+Ist quality
C B A quality b % %c1-5
% than % than 5 1-2
t/ha t/ha than % % t/ha than than % %
c1-5 c1-5b1 % than
t/ha % Mx1b1- t/ha % than than c1 Mx1 t/ha % than than
Mx2
2 b1 c1b1-2 c1 Mx1
a1 90.8 103.7 76.0 83.7
b1a2 85.6 97.7 70.0 81.8 88.2 100.0 73.0 73.0 82.8 100.0 100.0
c1 c1xb1 88.2 100.7 73.0 82.8
a1 90.2 103.0 73.2 81.2 87.6 100.0 73.0 71.6 81.7 100.0 73.5 67.6
b2 a2 83.8 95.7 67.1 80.1 87.0 98.6 73.0 70.2 80.7 96.2 100.0
c1xb2 87.0 99.3 70.2 80.7
B average for
87.6 100.0 71.6 81.7 87.6 99.3 73.0 71.6 81.7 98.1 100.0
a1
a1 140.5 104.6 106.9 76.1
b1 a2 128.9 95.8 95.6 74.2 134.7 100.0 111.5 101.3 75.2 100.0 138.8
c2 c2xb1 134.7 100.3 101.3 75.2
a1 136.8 101.9 97.0 70.9 134.3 153.3 111.9 97.4 72.5 136.0 100.0 92.0
b2 a2 131.0 97.5 90.0 68.7 133.9 99.4 112.3 93.5 69.8 92.3 133.2
c2xb2 133.9 99.7 93.5 69.8
B average for
134.3 100.0 97.4 72.5 134.3 99.7 111.9 97.4 72.5 96.2 136.0
a2
a1 129.6 106.1 114.2 88.1
b1 a2 116.2 95.2 100.2 86.2 122.9 100.0 101.7 107.2 87.2 100.0 146.8
c3 c3xb1 122.9 100.7 107.2 87.2 122.1 139.4 101.8 103.3 84.6 144.3 106.1 97.5
a1 126.7 103.8 104.3 82.3
b2 a2 115.9 94.9 94.3 81.4 121.3 98.7 101.8 99.3 81.9 92.6 141.5
c3xb2 121.3 99.3 99.3 81.9
372
Factor
Average production for:
experimental
Factor A Factor B Factor C
E+Ist % c1- E+Ist quality E+Ist quality
C B A quality b % %c1-5
% than % than 5 1-2
t/ha t/ha than % % t/ha than than % %
c1-5 c1-5b1 % than
t/ha % Mx1b1- t/ha % than than c1 Mx1 t/ha % than than
Mx2
2 b 1 c b
1 1-2 c1 Mx1
B average for
122.1 100.0 103.3 84.6 122.1 99.3 101.7 103.3 84.6 96.4 144.3
a3
a1 145.2 106.9 129.9 89.5
b1 a2 129.2 95.1 113.4 87.8 137.2 100.0 113.6 121.7 88.7 100.0 166.7
c4 c4xb1 137.2 101.0 121.7 88.7
a1 139.8 102.9 118.0 84.4 135.8 155.0 113.2 117.2 86.3 163.7 120.3 110.7
b2 a2 129.0 95.0 107.1 83.0 134.4 97.9 112.8 112.6 83.8 92.5 160.4
c4xb2 134.4 99.0 112.6 83.8
B average for
135.8 100.0 117.2 86.3 135.8 99.0 113.2 117.2 86.6 96.3 163.7
a4
a1 126.5 105.4 106.8
84.4
b1
a2 115.0 95.8 82.4 120.8 100.0 100.0 100.8 81.1 100.0 138.1
94.8
b1 120.8 100.7 100.8
83.4
c5 a1 123.4 102.8 98.1
79.5
b2
(Mx1) a2 114.9 95.7 89.6
78.0 119.2 98.7 100.0 93.9 79.6 93.2 133.8 120.0 137.0 100.0 97.4 81.2 136.0 100.0 91.8
b2 119.2 99.3 93.9
78.8
a1 125.0 104.2 102.5
82.0 * * * * * * *
a2 115.0 95.8 92.2
80.2 * * * * * * *
Mx1 120.0 100.0 97.4 81.2 120.0 99.3 100.0 96.5 80.4 95.7 134.8
b1 * * * * * 131.6 100.0 100.7 110.1 83.7 100.0 150.8
c6 b2 * * * * * 129.9 98.7 99.4 101.8 78.4 92.5 145.0
(Mx2 a1 136.4 104.4 111.7 81.9 * * * * * * * 130.7 149.2 108.9 105.9 81.0 147.9 108.7 100.0
*
a2 125.0 95.6 100.1 80.1 * * * * * * *
Mx2 130,7 100,0 105.9 81.0 130.7 99.3 100.0 105.9 81.0 96.2 150.0
In table 4 there are presented the statistical under the impact of the interaction between the
results of production differences significances experimental factors.
Table 4. Unilateral and interaction impact of experimental factors upon the production of some non-determined growth
tomato hybrids, cultivated on mineral substrate in industrial greenhouses
Average production Relative production Difference
Variant Significance
(t/ha) (%) (±t/ha)
1. Unilateral impact of the hybrid upon tomato production
a2-a1 114.95 124.95 91.99 -10.00 000
DL 5% = 1.60 DL 1% = 2.42 DL 0.1%= 3.89
2. Unilateral impact of the fertigation system upon tomato production
b2-b1 119.15 120.75 98.68 -1.60 -
DL 5% = 2.69 DL 1% = 3.71 DL 0.1% = 5.11
3. Unilateral impact of the flower bind improvement method upon tomato production
c2-c1 134.30 87.60 153.31 46.70 ***
c3-c1 122.10 87.60 139.38 34.50 ***
c4-c1 135.80 87.60 155.02 48.20 ***
c5-c1 119.96 87.60 136.94 32.36 ***
c3-c2 122.10 134.30 90.92 -12.20 000
c4-c2 135.80 134.30 101.12 1.50 -
c5-c2 119.96 134.30 89.32 -14.34 000
c4-c2 135.80 134.30 101.12 1.50 -
c5-c3 119.96 122.10 98.25 -2.14 -
c5-c4 119.96 135.80 88.33 -15.84 000
DL 5% = 2.60 DL 1% = 3.52 DL 0.1% = 4.71
373
(t/ha) (%) (±t/ha)
4. Interaction impact between different hybrids and the same or different fertigation systems upon tomato
production
a2b1-a1b1 114.97 126.53 90.87 -11.55 000
a2b2-a1b2 114.93 123.38 93.15 -8.45 000
a2b2-a1b1 114.93 126.53 90.83 -11.60 000
DL 5% = 3.13 DL 1% = 4.41 DL 0.1% = 6.32
5. Interaction impact between the same hybrid and different fertigation systems upon tomato production
a1b2- a1b1 123.38 126.53 97.51 -3.15 -
a2b2- a2b1 114.93 114.97 99.96 -0.05 -
DL 5% = 3.81 DL 1% = 5.24 DL 0.1% = 7.22
6. Interaction impact between the same hybrid and different flower bind improvement methods upon tomato
production
a1c2- a1c1 138.65 90.50 153.20 48.15 ***
a1c3- a1c1 128.15 90.50 141.60 37.65 ***
a1c4- a1c1 142.50 90.50 157.46 52.00 ***
a1c5- a1c1 124.97 90.50 138.08 34.47 ***
a1c3- a1c2 128.15 138.65 92.43 -10.50 000
a1c4- a1c2 142.50 138.65 102.78 3.85 *
a1c5- a1c2 124.97 138.65 90.13 -13.68 000
a1c4- a1c3 142.50 128.15 111.20 14.35 ***
a1c5- a1c3 124.97 128.15 97.52 -3.18 -
a1c5- a1c4 124.97 142.50 87.70 -17.53 000
a2c2- a2c1 129.95 84.70 153.42 45.25 ***
a2c3- a2c1 116.05 84.70 137.01 31.35 ***
a2c4- a2c1 129.10 84.70 152.42 44.40 ***
a2c5- a2c1 114.95 84.70 135.71 30.25 ***
a2c3- a2c2 116.05 129.95 89.30 -13.90 000
a2c4- a2c2 129.10 129.95 99.35 -0.85 -
a2c5- a2c2 114.95 129.95 88.46 -15.00 000
a2c4- a2c3 129.10 116.05 111.25 13.05 ***
a2c5- a2c3 114.95 116.05 99.05 -1.10 -
a2c5- a2c4 114.95 129.10 89.04 -14.15 000
DL 5% = 3.68 DL 1% = 4.98 DL 0.1% = 6.66
7. Interaction impact between the same fertigation system and different flower bind improvement methods
upon tomato production
b1c2- b1c1 134.70 88.20 152.72 46.50 ***
b1c3- b1c1 122.90 88.20 139.34 34.70 ***
b1c4- b1c1 137.20 88.20 155.56 49.00 ***
b1c5- b1c1 120.75 88.20 136.90 32.55 ***
b1c3- b1c2 122.90 134.70 91.24 -11.80 000
b1c4- b1c2 137.20 134.70 101.86 2.50 -
b1c5- b1c2 120.75 134.70 89.64 -13.95 000
b1c4- b1c3 137.20 122.90 111.64 14.30 ***
b1c5- b1c3 120.75 122.90 98.25 -2.15 -
b1c5- b1c4 120.75 137.20 88.01 -16.45 000
b2c2- b2c1 133.90 87.00 153.91 46.90 ***
b2c3- b2c1 121.30 87.00 139.43 34.30 ***
b2c4- b2c1 134.40 87.00 154.48 47.40 ***
b2c5- b2c1 119.17 87.00 136.97 32.17 ***
b2c3- b2c2 121.30 133.90 90.59 -12.60 000
b2c4- b2c2 134.40 133.90 100.37 0.50 -
b2c5- b2c2 119.17 133.90 89.00 -14.73 00
b2c4- b2c3 134.40 121.30 110.80 13.10 ***
b2c5- b2c3 119.17 121.30 98.24 -2.13 -
374
(t/ha) (%) (±t/ha)
b2c5- b2c4 119.17 134.40 88.67 -15.23 000
DL 5% = 3.68 DL 1% = 4.98 DL 0.1% = 6.66
8. Interaction impact between different fertigation systems and the same or different flower bind
improvement methods upon tomato production
b2c1- b1c1 87.00 88.20 98.64 -1.20 -
b2c2- b1c2 133.90 134.70 99.41 -0.80 -
b2c3- b1c3 121.30 122.90 98.70 -1.60 -
b2c4- b1c4 134.40 137.20 97.96 -2.80 -
b2c5- b1c5 119.17 120.75 98.69 -1.58 -
b2c2- b1c1 133.90 88.20 151.81 45.70 ***
DL 5% = 4.25 DL 1% = 5.80 DL 0.1% = 7.84
9. Interaction impact between different hybrids and the same or different flower bind improvement methods
upon tomato production
a2c1- a1c1 84.70 90.50 93.59 -5.80 00
a2c2- a1c2 129.95 138.65 93.73 -8.70 000
a2c3- a1c3 116.05 128.15 90.56 -12.10 000
a2c4- a1c4 129.10 142.50 90.60 -13.40 000
a2c5- a1c5 114.95 124.97 91.98 -10.02 000
a2c2- a1c1 129.95 90.50 143.59 39.45 ***
DL 5% = 3.65 DL 1% = 5.04 DL 0.1% = 6.99
10. Interaction impact between the same hybrid and the same fertigation system and different flower bind
improvement methods upon tomato production
a1b1c2- a1b1c1 140.50 90.80 154.74 49.70 ***
a1b1c3- a1b1c1 129.60 90.80 142.73 38.80 ***
a1b1c4- a1b1c1 145.20 90.80 159.91 54.40 ***
a1b1c5- a1b1c1 126.53 90.80 139.35 35.73 ***
a1b1c3- a1b1c2 129.60 140.50 92.24 -10.90 000
a1b1c4- a1b1c2 145.20 140.50 103.35 4.70 -
a1b1c5- a1b1c2 126.53 140.50 90.06 -13.97 000
a1b1c4- a1b1c3 145.20 129.60 112.04 15.60 ***
a1b1c5- a1b1c3 126.53 129.60 97.63 -3.07 -
a1b1c5- a1b1c4 126.53 145.20 87.14 -18.67 000
a2b2c2- a2b2c1 131.00 83.80 156.32 47.20 ***
a2b2c3- a2b2c1 115.90 83.80 138.31 32.10 ***
a2b2c4- a2b2c1 129.00 83.80 153.94 45.20 ***
a2b2c5- a2b2c1 114.93 83.80 137.15 31.13 ***
a2b2c3- a2b2c2 115.90 131.00 88.47 -15.10 000
a2b2c4- a2b2c2 129.00 131.00 98.47 -2.00 -
a2b2c5- a2b2c2 114.93 131.00 87.74 -16.07 000
a2b2c4- a2b2c3 129.00 115.90 111.30 13.10 ***
a2b2c5- a2b2c3 114.93 115.90 99.17 -0.97 -
a2b2c5- a2b2c4 114.93 129.00 89.10 -14.07 000
DL 5% = 5.20 DL 1% = 7.05 DL 0.1% = 9.42
11. Interaction impact between the same hybrid and different fertigation systems and the same flower bind
improvement method upon tomato production
a1b2c1- a1b1c1 90.20 90.80 99.34 -0.60 -
a1b2c5- a1b1c5 123.40 126.53 97.52 -3.13 -
a2b2c2- a2b1c2 131.00 128.90 101.63 2.10 -
a2b2c5- a2b1c5 114.93 114.97 99.97 -0.03 -
DL 5% = 6.01 DL 1% = 8.20 DL 0.1% = 11.09
12. Interaction impact between different hybrids and the same fertigation system and the same flower bind
improvement method upon tomato production
a2b1c1- a1b1c1 85.60 90.80 94.27 -5.20 -
a2b1c2- a1b1c2 128.90 140.50 91.74 -11.60 000
375
(t/ha) (%) (±t/ha)
a2b1c3- a1b1c3 116.20 129.60 89.66 -13.40 000
a2b1c4- a1b1c4 129.20 145.20 88.98 -16.00 000
a2b1c5- a1b1c5 114.97 126.53 90.86 -11.57 000
a2b2c1- a1b2c1 83.80 90.20 92.90 -6.40 0
a2b2c2- a1b2c2 131.00 136.80 95.76 -5.80 0
a2b2c3- a1b2c3 115.90 126.70 91.48 -10.80 000
a2b2c4- a1b2c4 129.00 139.80 92.27 -10.80 000
a2b2c5- a1b2c5 114.93 123.40 93.14 -8.47 00
DL 5% = 5.60 DL 1% = 7.68 DL 0.1% = 10.50
Analysing the significance of the production under the influence of c2 (stimulation with
differences from the table the following Tomato-Stim) being quantity wise superior but
conclusions emerge: not quality wise (table 3);
Point 1 - the unilateral influence of the hybrid Analysing points 4-12 from table 4 we see that
on the production - shows that the obtained based on the bi- or tri-factorial interactions,
production from the two hybrids have a there is a very big diversity in the significance
statistical assurance, the significance of the of the differences in production, that include all
production differences between Marissa F1 (a2) degrees of assessment (very significantly
and Noralee F1 (a1) being considerably positive or negative, distinctively significant
negative, meaning that the Noralee F1 (a1) positive or negative and significant positive or
hybrid has superior production qualities negative). This proves the intensity of the
compared to Marissa F1 (a2), 125 t/ha and unilateral influences or the interactions of the
respectively 115 t/ha, but quality as well, as the experimental factors on the productions
data from table 3 shows; obtained from a quantitative angle.
Point 2 - the unilateral influence of the
fertigation system on the production - shows CONCLUSIONS
that the productions obtained under the
influence of the two fertigation systems do not The productions following the influences of
have a statistical assurance, having no some experimental factors, such as the
significant production differences, which fertigation system with different types of
proves that both fertigation systems can be used modern chemical instant fertilizers,
because the difference between the two is very administered interacting bi-factorial with the
low in relation to the average production per cultivated hybrid, have proved the complexity
hectare (119.2 t/ha respectively 120.8 t/ha); of the interactions (tri-factorial) with the
Point 3 - the unilateral influence of the improved flower binding methods by
improved flower binding method on the differentiating them quantitatively and
production - shows that the production obtained qualitatively in a tomato culture performed on a
under the influence of method c2 (stimulation substrate of mineral cotton.
with Tomato-Stim), c3 (mechanical pollination) The diversity of factor C graduation (Improved
and c4 (natural pollination via bumble-bees) flower binding method) over the tomatoes
have statistical assurance, the significance of production, have determined influences over
the production differences compared to c1 the quantity as well as quality types, namely the
(unstimulated and non-pollinated Mt) being ones which through their natural influence have
essentially positive in all cases, which shows contributed to a better pollination and
that all used methods are beneficial for the fertilization, thus a better flower binding. The
improvement of flower binding compared to same as for Carlos de Melo e Silva Neto et al.
the controlled variant; also significance of the (2013) results, our results show that native bees
difference between c3 (mechanical pollination) buzz-pollinate tomato flowers, increasing the
and c2 (stimulation with Tomato-Stim) is pollen load on their stigma and consequently
essentially negative, which proves that the fruit production and quality.
production has statistical assurance, namely
376
Due to the influence of c4 graduation The influence of both graduation of the
(bumblebees natural pollination) and c2 fertigation systems (Yara and Haifa Chemicals)
(Tomato-Stim) the biggest average tomato are substantiated in the very close quantitative
production within the experiment was of 135.8 production and from a statistic point of view
t/ha and respectively 134.3 t/ha, but not the best were not covered, because there were no
quality in both graduations, as Extra+Ist quality significant production differences.
production. This is due to the fact that the Noralee F1 and Marissa F1 hybrids are
pollen dose added to the stigmas of tomato valuable both due to the quantitative level of
flowers (with the help of bumblebees) lead to a production as well as the value of Extra+Ist
good pollination (meaning a good production quality production, these ranging without
increase) and also, if we are to consider the taking into account c1 - Mt, throughout the
other aspects, an increase in the number of following intervals:
fertilized eggs, which also mean an increase in - Noralee F1: 126.7-145.2 t/ha, of which 97.3-
the production of seeds in the fruits. Related to 129.9 t/ha Extra+Ist quality production,
this, studies developed by Tanksley (2004) and meaning 70.9-89.5%;
Paran et al. (2007) have shown that the number - Marissa F1: 115.9-131.0 t/ha, of which 90.0-
of seeds in development in tomato fruits 113.4 t/ha Extra+Ist quality production,
influences the activity of the fw 2.2 gene, meaning 68.7-87.8%.
which is responsible for the production of Based on the conclusions following the
stimuli for the ovary walls growth and fruit performed research, the recommendation is:
formation (quoted by de Carlos de Melo e Silva - The cultivation of Noralee F1 hybrid;
Neto et al., 2013). - The use of both fertilizers Yara and Haifa
It has been observed that under the c2 Chemicals via fertigation;
graduation (Tomato-Stim) the production on - The use of mechanical means or bumblebees
second place in quantity size is on the last place (Biobest or Natupol) to improve the tomato
regarding the quality Extra+Ist quality flower binding and to obtain superior quality
production. fruits.
The Extra+Ist quality production, from a
percentage aspect at its highest was obtained REFERENCES
under the c4 graduation (bumblebees natural
pollination) - 86.3%, followed chronologically Bispo dos Santos S.A., Roselino A.C., Hrncir M., Bego
L.R., 2009. Pollination of tomatoes by the stingless
by c3 (mechanical pollination) - 84.6%. bee Melipona quadrifasciata and the honey bee A.
According to Carlos de Melo e Silva Neto et mellifera (Hymenoptera, Apidae). Genet. Mol. Res.
al., 2013, it has been shown that in greenhouses 8: 751-757.
Melipona quadrifasciata bees, gains in fruit Buchmann S.L., Cane J.H., 1989. Bees assess pollen
production reached 15% (Bispo dos Santos et returns while sonicating Solanum flowers. Oecologia
81: 289-294.
al., 2009), while with B. impatiens, gains Carlos de Melo e Silva Neto, Flaviana Gomes Lima,
reached 50% in fruit mass and up to the double Bruno Bastos Gonçalves, Leonardo Lima Bergamini,
in the number of seeds (Morandin et al., Barbara Araújo Ribeiro Bergamini, Marcos Antônio
2001a). da Silva Elias, Edivani Villaron Franceschinelli,
The classification from a quality point of view 2013. Native bees pollinate tomato flowers and
increase fruit production, Journal of Pollination
of the production on the first two places, Ecology, 11 (6), 2013, pp 41-45.
productions under the influence of c4 Harder L.D., 1990. Behavioral responses by bumblebees
(bumblebees natural pollination) and c3 to variation in pollen availability. Oecologia 94: 244-
(mechanical pollination), is based on the 246.
natural pollination phenomenon which Horgoş A., Oglejan Doina., Kondor F., Becherescu Al.,
2002. On the influence of Kemira-type fertilising on
determines the growth and development on the the quantitative and qualitative levels of tomato
plant of non-parthenocarpic fruits, with yields in unheated solaria. Scientifical Works B
outstanding physical (size, weight and color series (XLV).
etc.), chemical and organoleptic (flavor, smell Horgoş A., Oglejan D., Bulboacă T., Becherescu A.,
etc.) characteristics. 2005. Researches regarding the cultivation of
tomatoes with growth determined on mineral and
organic stratum in industrial hothouses warned with
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Agricultural Sciences and Veterinary Medicine, Cluj- Pollination Ecology 11 (5): 33-40.
Napoca, Vol.62, HORTICULTURE Book Series: Olmstead R.G., Palmer J.D., 1997. Implications for the
Pages: 136. phylogeny, classification and biogeography of
Kevan P.G., Straver W.A., Offer M., Laverty T.M., Solanum from cpDNA restriction site variation.
1991. Pollination of greenhouse tomatoes by Systematic Botany 22: 19-29.
bumblebees in Ontario. Proceedings of the Paran I, van der Knaap E., 2007. Genetic and molecular
Entomological Society of Ontario 122: 15-19. regulation of fruit and plant domestication traits in
Morandin L.A., Laverty T.M., Kevan P.G., 2001a. tomato and pepper. Journal of Experimental Botany
Bumblebee (Hymenoptera: Apidae) activity and 58: 3841-3852.
pollination levels in commercial tomato greenhouses. Shelly T.E., Villalobos E., OTS-USAP, 2000. Buzzing
Journal of Economic Entomology 94 (2): 462-467. bees (Hymenoptera: Apidae, Halictidae) on Solanum
Morandin L.A., Laverty T.M., Kevan P.G., 2001b. Effect (Solanaceae): floral choice and handling time track
of bumblebee (Hymenoptera: Apidae) pollination pollen availability. Florida Entomologist 83 (2): 180-
intensity on the quality of greenhouse tomatoes, 187.
Journal of Economic Entomology 94: 172-9 Tanksley S.D., 2004. The genetic, developmental, and
Morse A., 2009. Floral scent and pollination of molecular basis of fruit size and shape variation in
greenhouse tomatoes. Master of Science Thesis. The tomato. The Plant Cell 16:181-189.
University of Guelph: 1-3, 101-102. Teppner H., 2005. Pollinators of tomato, Solanum
Nunes-Silva P., Hnrcir M., Shipp L., Imperatriz-Fonseca lycopersicum (Solanaceae), in Central Europe.
V.L., Kevan P.G., 2013. The behaviour of Bombus Phyton 45(2): 217.
impatiens (Apidae, Bombini) on tomato
(Lycopersicon esculentum Mill., Solanaceae)
378
RESEARCH ON THE EFFECT OF CLIMATE CHANGE

ON THE PRODUCTION CAPACITY OF SOME SWEET POTATOES
VARIETIES (IPOMOEA BATATAS) CULTIVATED ON PSAMOSOILS
IN THE SOUTHERN AREA OF ROMANIA
Aurelia DIACONU, Alina-Nicoleta PARASCHIV, Reta DRĂGHICI,

Mihaela CROITORU, Milica DIMA
Research and Development Station for Plant Culture on Sands, Dăbuleni,

217 Petre Baniţă, Dolj County, Romania
Abstract
Between 2015 and 2017, at the Research and Development Station for Plant Culture on Sands Dăbuleni, were studied
three varieties of sweet potato to determine the influence of environmental factors on some plant metabolic processes
and, consequently, on the production achieved. The varieties studied were Yulmi, Juhwangmi and KSP 1. The results
revealed the influence of the variety, the planting season and the climatic conditions on the development of
physiological processes, their intensity influencing the quantity and quality of production. The best results on the
production were recorded at the variants cultivated under the climatic conditions of the 2016 year, detaching the
variant cultivated with the Juhwangmi variety, with an average production of 43.6 tons. At Yulmi and KSP 1 varieties it
was obtained the highest production at variants grown in the first epoch of 2016 year (May 10-15), respectively 34.3
tons at Yulmi variety and 31.7 tons at KSP 1 variety.
Key words: climate change, foliar transpiration, photosynthesis, production, sweet potato.
INTRUDUCTION genus Ipomoea on the basis of the stigma shape

and the surface of the pollen grains. Therefore,
Sweet potato (Ipomoea batatas L.), the seventh the name was changed to Ipomoea batatas (L.)
most important food crop after wheat, rice, Lam.
maize, potato, barley and cassava, is a staple Sweet potato was originally a herbaceous
food in many developing countries of the perennial but was domesticated as an annual
tropics and sub-tropics, and also serves as and grows best in moderately warm tempe-
animal feed and raw material for several food rature of 21-26oC. It requires light textured soil
and feed based industries (Ray and Ravi, with the optimum pH of 5.5-6.5 for good
2005). growth of the crop. At low pH sweet potato
This New World crop has high biological suffers aluminum toxicity.
efficiency of converting solar energy into Sweet potato is sensitive to alkaline and saline
edible energy (152 MJ/ha/day) in the form of conditions (Dasgupta et al., 2006; Mukherjee et
tuberous (storage) roots and could al., 2006).
be the food for the ever growing human A well distributed rainfall of 75-150 cm is
population in future ( Ray and Tomlins, 2010). favourable for its cultivation. It can tolerate
Asia leads in area (60.75%) and production drought to some extent but cannot withstand
(86.89%) of sweet potato in the world. The water logging. It requires plenty of sun shine,
world average sweet potato root yield was whereas shade causes reduction in yield.
13,729 kg/ha. However, the highest However, sweet potato is grown as intercrop
productivity of 19,634 kg/ha was found in Asia under plantation/orchard crops with the motto
(FAOSTAT, 2008). of profit maximization and crop intensification
Sweet potato was first described by Linnaeus (Nedunchezhiyan et al., 2007).
as Convolvulus batatas in 1753. However, in Excess of rainfall and cloudy conditions
1791 Lamarck classified this species within the encourage vine growth and reduce storage root
379
yield. Dry season storage root yields were PRO + portable device were determined the
higher than rainy season yields diurnal variation of photosynthesis and foliar
(Nedunchezhiyan and Byju, 2005). transpiration rate in different vegetation
In Romania was introduced in 1954, and the phenophases. Determinations have also been
result of the experiments performed by Maier I. made on the commercial production obtained
was further amplified (Florescu, Maxim, during the three years of study, depending on
Dragotă, Ciofu, Popescu) and allowed the the variety and planting epoch.
acclimatization of this new species, as well as
the establishment of sequences on the way of RESULTS AND DISCUSSIONS
multiplication, planting periods, crop areas. In
the current context of climate change, the From the data presented in Table 2 it can be
importance of sweet potato cultivation lies both estimated that the three years under study
in the possibility of expanding culture in the (2015-2017) are different from the climatic
areas where the potato is degenerating point of view.
(Ruxandra Ciofu et al., 2003) and the need to Analyzing the air temperature values, during
diversify the vegetable variety with less known the sweet potato growing season (May-
species but with a nutritional value which September), there is a tendency to increase the
would make efficient use of the pedoclimatic average monthly temperature over the three
conditions in southern Oltenia. years of study. It is also noted that the monthly
average temperature of each year was higher
MATERIALS AND METHODS than the average multiannual temperature. On
the other hand, the maximum temperatures also
At RDSPCS Dabuleni, 3 potato varieties were increased steadily, in 2017 values exceeding
studied between 2015-2017 to determine the 40oC in all summer months (June-August).
influence of environmental factors on plant Heat temperatures have blocked physiological
metabolic processes and, consequently, on the processes in sweet potato plants, negatively
production achieved. influencing the development of vegetation
The varieties studied were Yulmi, Juhwangmi phenophases and the accumulation of starch in
and KSP 1. The experimental variants were tuberous roots.
cultivated according to the technology, namely The atmospheric precipitations, as well as their
in field with ridges, protected with transparent relationship with foliar transpiration values, are
polyethylene mulch, in three planting epochs, of particular importance for the southern
at an interval of 10-15 days between epochs Oltenia sweet potato culture.
(Table 1), and water was provided by irrigation The precipitation recorded between May and
using the drip irrigation method. September showed values between 227.8 mm
in 2016 and 330.8 mm in 2015, unevenly
Table 1. Experimental variants distributed over the vegetation period and with
Experimental variants variations from one month to the next.
Yulmi Planting I (10-15 May) Throughout the analyzed period there is a
Variety Juhwangmi epoch II (20-25 May) significant reduction in the amount of
KSP 1 III (01-10 June) precipitation in July, August and September.
Although sweet potato is a drought-resistant
For the analysis of the climatic conditions species, heat temperatures correlated with a
which influenced the sweet potato plants in the major lack of rainfall have led to droughts and
experimental field, were used the have made it absolutely necessary the irrigation
meteorological data recorded at the RDSPCS of crop.
Dabuleni meteorological station. With the LC
380
Table 2. The variation of the main climatic elements during the growing season of sweet potatoes
in the period 2015-2017
Year Climatic element Month
Medium temperature (oC) 19.2 20.5 24.8 24.34 20.08
Minimum temperature (oC) 8.6 10.2 12.5 12.9 7.8
2015 Maximum temperature (oC) 30.2 36.1 39.2 37.6 37.3
Precipitations (mm) 52.4 134.2 11 48.4 84.8
Atmospheric relative humidity (%) 73.04 73.77 62.92 68.21 77.30
Medium temperature (oC) 16.8 23.6 24.8 23.5 20.4
Minimum temperature (oC) 5.5 11 11.4 16.1 5.1
2016 Maximum temperature (oC) 32.9 37.3 38 38 34.1
Precipitations (mm) 104.4 53.2 31.6 1 37.6
Atmospheric relative humidity (%) 82.22 72.68 65 68.3 71
Medium temperature (oC) 17.8 24 24.8 24.8 20.21
Minimum temperature (oC) 4.7 12.9 13.3 11 6.7
2017 Maximum temperature (oC) 31.4 41.2 40.8 40.4 36.9
Precipitations (mm) 78.6 17.4 120.8 28,8 18.2
Atmospheric relative humidity (%) 77 67.05 65.01 63 66
Multiannual medium temperature 16.8 21.6 23.1 22.4 17.8
(1956-2016)
Precipitations, multiannual total 62.12 69.30 53.15 37.28 47.83
(1956-2016)
In the context of climate change, the results photosynthetic yields beeing presented only the
regarding physiological processes on sweet drought-resistant varieties (in the present case
potato plants grown on sandy soils in southern the Yulmi variety).
Oltenia highlighted the following aspects:
Both the rate of photosynthesis and the foliar Table 3. Variation of photosynthesis at sweet potato in
transpiration rate varied depending on the the phenophase of intense growth of strains according to
variety, planting epoch and the culture year
variety, the vegetation phenophase analyzed,
the planting epoch and the climatic conditions Photosynthesis rate
(µmol CO2/m2/s) Average
present at the time of the measurements. From The first
The The /
the data presented in Table 3 it can be noticed Variety Year
epoch
second
epoch
third
epoch
Variety
that in the phase of intense growth of the May 10-

15
May 20- June
strains, there were no photosynthetic 25 01-10
2015 22.32 16.90 16.02 18.41
differences between the three studied varieties, Yulmi 2016 18.33 13.26 12.72 14.77
the average values of this physiological process 2017 24.34 29.42 29.33 27.69
Average/Epoch 21.66 19.86 19.35 20.29
being between 20.29 μmol CO2/m2/s at the 2015 19.32 19.34 15.77 18.14
Yulmi variety, respectively 22.55 μmol Juhwangmi 2016 18.08 16.59 17.8 17.49
CO2/m2/s at the KSP 1 variety. As regards the 2017
Average/Epoch
21.34
19.58
25.29
20.40
31.83
21.80
26.15
20.59
influence of the planting epoch on the 2015 22.20 15.95 25.43 21.19
photosynthesis in this vegetation phenophase, KSP 1 2016 15.28 20.82 17.82 17.97
2017 28.96 29.18 27.33 28.49
the results are easily differentiated according to Average/Epoch 22.14 21.98 23.52 22.55
the variety as follows: Yulmi variety has
presented the best results at cultivation in first The intensity of foliar transpiration is directly
epoch (21.66 μmol CO2/m2/s); the Junhwangmi related to the temperature and hydrological
and KSP 1 varieties showed the highest deficit of atmospheric air. By decreasing the
photosynthetic yields at cultivation in the third degree of saturation of air in water vapor, the
epoch (21.8 μmol CO2/m2 /s, respectively 23.52 water deficit and the intensity of foliar
μmol CO2/m2/s). This can be explained by the transpiration increase. Table 4 presents the
fact that at the plants cultivated in the first variation of this physiological process in the
epoch this vegetation phenophase coincides phenophase of intense growth of strains
with the harshest period from the climatic point according to variety, planting epoch and culture
of wiew, high year. The most intensely has sweated the KSP
381
1 variety, with an average value over the three- Table 5. Variation of photosynthesis and foliar
year study of 5.47 mmol H2O/m2/s, but transpiration at sweet potato in the root tuberosis
phenophase according to variety, planting epoch and the
evaporated water was efficiently used because culture year
this variety also recorded the highest average
Photosynthesis rate (µmol
value of photosynthesis. Regarding the CO2/m2/s) Average
influence of the planting epoch on the intensity The first
The The /
Variety Year second third Year
of foliar transpiration (Table 4), it is observed epoch
epoch epoch
May 10-
that the highest values were recorded in the 15
May 20- June
25 01-10
plants grown in the third epoch, indifferent to 2015 10.17 10.6 13.89 11.55
the variety. If we analyze the year's influence Yulmi 2016 17.77 14.76 10.4 14.31
2017 15.68 24.09 27.08 22.28
on transpiration, it is noticed that the highest Average/Epoch 14.54 16.48 17.12 16.04
values were recorded in the climatic conditions 2015 5.36 8.87 12.89 9.04
of 2017, this year being one of the droughts. Juhwangmi 2016 21.18 15.69 13.91 16.92
2017 25.48 21.05 33.07 26.53
Average/Epoch 17.34 15.20 19.95 17.5
Table 4. Variation of foliar transpiration at sweet potato 2015 6.64 8.91 11.2 8.91
in the phenophase of intense growth of strains according KSP 1 2016 19.19 12.71 14.75 15.55
to variety, planting epoch and the culture year 2017 22.93 19.44 27.48 23.28
Average/Epoch 16.25 13.68 17.81 15.91
Foliar transpiration rate Foliar transpiration rate
(mmol H2O/m2/s) Average / 2
(mmol H2O/m /s) Average
The The The Year The The /
The first
Variety Year first second third Variety Year second third Year
epoch
epoch epoch epoch epoch epoch
May 10-
May May June May 20- June
15
10-15 20-25 01-10 25 01-10
2015 6.56 3.90 4.67 5.04 2015 1.40 2.35 2.21 1.98
Yulmi 2016 3.65 3.64 2.45 3.24 Yulmi 2016 3.83 4.06 2.41 3.43
2017 3.46 5.72 9.67 6.28 2017 3.12 3.91 7.65 4.89
Average/Epoch 4.55 4.42 5.59 4.85 Average/Epoch 2.78 3.44 4.09 3.43
2015 6.11 3.18 5.25 4,84 2015 1.34 2.13 1.76 1.74
Juhwangmi 2016 4.46 4.21 3.25 3.97 Juhwangmi 2016 4,33 4.24 2.63 3.73
2017 5.13 6.32 8.55 6.66 2017 4.27 5.04 6.53 5.28
2015 6.30 3.75 6.46 5.50 2015 1.35 1.33 2.01 1.56
KSP 1 2016 3.44 5.57 3.14 4.05 KSP 1 2016 4.49 2.43 1.81 2.91
2017 6.01 6.15 8.46 6.87 2017 4.55 5.42 5.69 5.22
In the root tuberosis phenophase, both Analyzing the production of commercial tubers
physiological processes studied have reduced obtained at 120 days after the sweet potato
their intensity as a result of the maturation planting in the experimental field, it note the
process of the plants (Table 5). importance of the variety, of planting epoch
Remarkable are the results obtained in and the climatic conditions of the three years of
experimental year 2017, in this vegetation phe- study on the results obtained (Table 6). The
nophase the values of photosynthesis and foliar best results on the production were recorded at
transpiration beeing unusually large (comprised the variants cultivated under the climatic
in the range 22.28-26.53 μmol CO2/m2/s, conditions of the 2016 year, detaching the
respectively 4.89-5.28 mmol H2O/m2/s). variant cultivated with the Juhwangmi variety,
Heat days, with temperatures exceeding 40oC with an average production of 43.6 tons.
in all the summer months (June to August), At Yulmi and KSP1 varieties it was obtained
determined an increase in the physiological the highest production at variants grown in the
processes of sweet potato plants towards the first epoch of 2016 year (May 10-15),
end of the vegetation period when the climatic respectively 34.3 tons at Yulmi variety and
conditions became optimal. 31.7 tons at KSP 1 variety. Analyzing the
This fact also had repercussions on the influence of the planting epoch on the
production, especially on the plants grown in production obtained, the best results were
the first epoch. obtained when the sweet potato planting was
done during May 20-25, with an average of
33.3 tons per hectare.
382
Table 6. Production results obtained at 120 days also recorded the highest CO2 accumulations in
after planting of sweet potatoes, depending on variety, the photosynthesis process.
year and planting epoch
The high photosynthetic yield places sweet
Epoch I Epoch II Epoch III Variety potato among the most productive vegetable
Variety Year average
10-15
May
20-25
May
10-15
June kg/ha
species grown on the sandy soils at Dabuleni.
2015 32200 26425 18550 25725
Although the climatic conditions were
different, notable commercial productions were
Yulmi 2016 34300 33250 11550 26366.67
obtained in all the studied years, quantitatively
2017 20800 25333.33 33400 26511,11
detaching the Juhwangmi variety. Regarding
Epoch average/variety 29100 28336.11 21166.67 26200.92
the planting epoch, the best results were
2015 40250 41300 30800 37450
obtained at plants grown between May 20-25
Juhwangmi 2016 48066.67 56000 26845 43637.22
(2nd epoch), with an average of 33.3 tons per
2017 24933.33 44000 43000 37311.11 hectare.
Epoch average/variety 37750 47100 33548.33 39466.11
2015 28233 22400 23800 24811 REFERENCES
KSP 1 2016 31733.33 25200 12810 23247.78
2017 17333.33 26093 14933.33 19453.22 Dasgupta M., Kole P.C., Sahoo M.R., Mukherjee A.,
Epoch average/variety 25766.55 24564.33 17181.11 22504
2006. Screening of sweet potato genotypes for
salinity stress. In: 14th Triennial Symp. Int. Soc. Trop.
Epoch average/
30872.18 33333.48 23965.37 Root Crops, 20-26 November 2006, CTCRI,
3 years
Thiruvanathapuram, India, pp. 166-167.
FAOSTAT, 2008. Current sweet potato production from
CONCLUSIONS selected different countries in different regions of the
world based on 2006 FAO survey. Available HTTP:
http://apps.fao.org.
The results obtained showed that sweet potato Mukherjee A., Naskar S.K., Edison S., Dasgupta M.,
cultivated in drip irrigation conditions, 2006. Response of orange flesh sweet potato
successfully tolerates the thermo-hydric stress genotypes to salinity stress. In: 14th Triennial Symp.
characteristic of the sandy soils area, the Int. Soc. Trop. Root Crops, 20-26 November 2006,
second planting epoch (May 20-25th) offering Central Tuber Crops Research Institute,
Thiruvanathapuram, India, pp. 151-152.
optimal conditions for the development of Nedunchezhiyan M., Byju G., 2005. Effect of planting
physiological processes in plants, regardless of season on growth and yield of sweet potato (Ipomoea
variety. Very high temperatures during the batatas L.) varieties. J. Root Crops 31 (2): 111-114.
vegetation period (which at the leaves level are Nedunchezhiyan M., Byju G., Naskar S.K., 2007. Sweet
with a few degrees higher than in the air), potato (Ipomoea batatas L.) as an intercrop in a
coconut plantation: Growth, yield and quality. J. Root
strong solar radiation, very low air humidity, Crops 33 (1): 26-29.
have acted as dehydrating forces of the foliar Ray R.C., Ravi V., 2005. Post-harvest spoilage of sweet
apparatus, increasing water losses through potato and its control measures. Crit. Rev. Food Sci.
foliar transpiration. Evaporated water was, Nutr. 35: 623- 644.
however, used efficiently, as varieties with Ray R.C., Tomlins K.I., 2010. Sweet Potato: Post-
harvest aspects in food, feed and industry. Nova
large water losses through foliar transpiration Science Publisher, Inc. New York.
383
384
THE INFLUENCE OF FOLIAR BIOACTIV TREATMENTS

ON TOMATOES SEEDLINGS
Elena DOBRIN, Evelina GHERGHINA, Gabriela LUŢĂ, Daniela BĂLAN

Abstract
Tomato (Lycopersicon esculentum), the world's most important vegetable species, are known for its sensitivity
regarding different stress factors such as heat, drought, lack of nutrients. Therefore, is very important to obtain
tomatoes seedlings with balanced growth and development, but especially with a strong root system and increased
capacity of adaptation to different conditions of stress. This paper presents the results of some researches related to
foliar bioactive substance treatment of tomatoes seedling, Rio Grande cultivar, with Spraygard 1%, Razormin 0.1%,
BAC Foliar Spray 0.3% and Bio Roots 0.2%. The treatments were performed in two distinct stages of development: at
one, respectively two weeks after the seedlings transplantation. Analysis of recorded data indicated that all variants
treated with bioactive substances are superiors comparing with the untreated control variant but Razormin 0.1%,
followed closely by Bio Roots 0.2% treatments showed the best results in obtaining of tomatoes seedlings with a strong
roots systems and a good development. These results are supported by an increased physiological activity.
Key words: growth, photosynthesis, root, transpiration.
INTRODUCTION synergistic effects of the interaction of several

factors when it is multiple stress (Pallavi et al.,
Abiotic stresses such as extremes temperature, 2012; Hasanuzzaman et al., 2013).
drought and salinity can reduce the yield of A biostimulator is an organic material that,
major crops and limit agricultural production when applied in small quantities, enhances
worldwide (Wang et al., 2003; Sharma et al., plant growth and development such that the
2012; Hasanuzzaman et al., 2012; response of the plant cannot be attributed to the
Hasanuzzaman et al., 2013). application of traditional nutrients (Shekhar
Different factors of abiotic and biotic stress Sharma et al., 2014)
determine in plants an avalanche of Different studies have shown that seaweed
physiological and biochemical changes, which extracts protect plants against a number of
begins with the perception of stress by the biotic and abiotic stresses and offers potential
specialized receptors and ends with the for field application. Further, seaweed extracts
resulting biomorphological expression as a are considered an organic farming out as they
result of this action. Therefore, the metabolism are environmentally safe for the health of
of the plants and the activity of some enzymes animals and humans (Khan et al., 2009).
is modified, known as the fact that all the Recent research showed that the use of red
chemical processes are catalyzed by enzymes. macroalgal on crop plants can generate
In plants, stress triggers a wide range of multiple benefits. It has been reported effects
responses, from gene expression and cell including enhanced rooting, enhanced
metabolism to changes in growth and photosynthetic activity, freezing, drought and
production, but when stress is severe, the plant salt tolerance, resistance to fungi, bacteria and
dies (Sharma et al., 2012). The response of virus, and higher crop and fruit yields (Shekhar
plants to stress is determined by a number of Sharma et al., 2014). Also, in recent years, a
factors, such as those that characterize the plant number of exogenous protectants, such as
(their genotype and stage of development) and proline, glycinebetaine, nitric oxide, silicon,
those that characterize stress (intensity, selenium, salicylic acid, and polyamines have
duration, repeatability, and the various been tested and found to be beneficial in
385
protecting plants against damage from MATERIALS AND METHODS
temperature extremes (Hasanuzzaman et al.,
2012). The experiment was installed into an
At this time, agricultural biostimulators experimental greenhouse of the Hortinvest
suppose different compounds with diverse Research Centre - USAMV Bucharest in April-
formulations of substances and other products, May, aimed to test the action of biofertilizers
such as microorganisms, trace elements, Razormin, BAC Foliar Spray and Bio Roots
enzymes, plant growth regulators and algal and of the universal adjuvant Spraygard on
extracts that are applied to plants or soils to growth and development of tomatoes seedlings,
regulate and enhance the crop’s physiological in order to be recommended to the seedlings
processes, in a very efficient manner (Shekhar producers as supportive treatment of the growth
Sharma et al., 2014). rate and to improve metabolism seedlings. This
In our country, similar research have been could lead to shortening their age with
made on tomato and pepper seedlings that have implications for reduction of production cost.
benefited from biofertilization with Cropmax, Spraygard is a complex product that acts as
Razormin, or of treatment with Spraygard safener, penetrant, dispersant, creates adhesion
(Chilom et al., 2000; Bălan et al., 2014; Dobrin of the treatment solutions on the leaves.
et al., 2014). The research on eggplant Spraygard adjuvant has an unique formula in a
seedlings reported that used of a different single coating based on the synthetic resin that
growth regulators (Razormin, Spraygard, BAC is "environmentally friendly" and the polymer
Foliar spray, Bio Roots) as foliar treatments, di-1-p-menthene and ethoxylatedalcohol by
induces biochemical, physiological and applying it on the plant and on its leaves forms
metabolic changes that have led to the a pellicle that persists 2 days up to 2 weeks,
improvement of seed growth and development having as a side effect the reduction of
but also to the improvement of the oxidative perspiration and, therefore, a better water
protection system of plant and attenuate the management within the plant. This fact causes
negative impact of heat stress (Bălan et al., the physiological chain reactions whose results
2018). are being expressed by increasing the plant
The present research reported were performed resistance to stress factors such as the drought
on tomatoes (Lycopersicum esculentum Mill.), and the cold. The effect of reducing
a main specie of vegetables grown in the field, transpiration recommends to apply the product
in the summer-autumn crop established by strictly on the leaves.
seedlings. Tomato seedlings are grown in Razormin is an environmentally friendly bio-
protected areas during April-May, when heat is stimulating product, which determines a
often installed, with maximum daily and rooting effect. Their chemical composition is
nocturnal temperatures well above optimum complex and balanced, so that induces mainly a
vegetation. It is imperative to apply a series of root system development, than the development
technological measures in order to accustom of vegetative part through cell division. It
seedlings with provided variable nonfatal stress contains free amino acids and polysaccharides,
that could exist in the place of cultivation and which stimulate the nutrients absorption,
to allow formation of "memory stress": the leading to the further development of plant.
correct and careful directing of the vegetation BAC Foliar is a foliar organic nutrient which
factors and the application of special treatments stimulates chlorophyll production in the leaves.
helping not only to form this memory of stress Bio Roots is a natural fertilizer that stimulates
but also supporting the growth and root growth, helping plants to form healthy and
development of plants (Sekara et al., 2012; vibrant root systems. It contains vitamins,
Mangrich and Saltveit, 2000; Knight et al., enzymes, organic and humic acids that
1996; Jennings and Saltveit, 1996). It were stimulate the growth of roots. It activates the
used different growth regulators as foliar growth of microorganisms in the root area and
treatments on tomatoes seedlings and the improves its development. This allows the
comparative results were studied. development of a strong root system and a
better adaptation of the plant to its
386
environment. At the same time it helps the RESULTS AND DISCUSSIONS
plant to cope with diseases and pathogenic
molds. The results of the analysis of the first stage (one
We established a monofactorial experiment week after the first treatment) are shown in
with 5 variants, considering application of Table 1.
bioactives substances Sparygard 1%, Razormin Since then tomato seedlings presented
0.1%, BAC Foliar Spray 0.3% and Bio Roots important nuances regarding growth as a result
0.2% on tomatoes seedlings (Rio Grande) in of the application of different treatments in the
two distinct stages: at one, respectively two sense that, in the non-fertilized V1 variant, the
weeks after the seedlings transplantation (April plants have recorded biometric parameters and
23 and 30). indicators far inferior to fertilized variants. The
The experimental variants were: V1-untreated plant height recorded an amplitude of 3.2 cm,
seedlings; V2-seedlings treated with 1% respectively, between 14.2 cm to V1 and 17.4
Spraygard; V3-seedlings trated with 0.1% cm to V4. The number of leaves varied
Razormin; V4-seedlings treated with BAC significantly, from 4.2 leaves formed at V1 to
Foliar Spray 0.3%; V5-seedlings treated with 5.2 leaves formed at V4. The leaf frequency is
Bio Roots 0.2%. relatively constant, of 0.3 leaf/cm PA to V1, V3
Sowing was made on April and because heat and V4 and 0.31 leaf/cm PA to V2.
and water were optimal provided, mass Table 1. Growth of tomatoes seedlings 7 days
emergence of seedlings occurred after 6 days after the first treatment
(April 11). The transplantation was done after 5 Variant Plants height No. of Leaves frequency
days into alveolar pallets (alveolar ø = 6 cm), in HPA leaves (no./cm HPA)
a professionally nutrient substrate (cm)
V1 14.2 4.2 0.30
KEKKILABP 75% +25% perlite. During the V2 15.4 4.8 0.31
growth period specific agrotechnics for V3 16.2 4.9 0.30
seedling production was applied: daily V4 17.4 5.2 0.30
ventilation, watering, weeding. The V5 16.0 4.8 0.30
temperature was kept at 24-26oC to 30oC at day
and 22-24oC at night. A treatment with CE
Bravo 0.2% was made in order to prevent
seedlings fall and also to avoid a downy mildew
attack.
Observations and measurements of plant
growth were made during the development of
experiments in different stages: a two and
respectively three weeks after transplantation
(at the end of the experiment), after 27 days to Figure 1. Influence of applied treatment on growth of
tomatoes seedlings 7 days after first treatments
the emergence, when most of seedlings have
reached the optimum for a succesfull planting.
Context analysis highlights V4 (treated with
Observations and measurements were made on
BAC Foliar Spray 0.3%) as the variant where
seedlings growth, as follow:
seedlings had the most balanced growth and the
biometric parameters of seedlings: plant
fact that the treatment applied since then has
height; the number of true leaves; weight of
exerted a decisive influence on the growth of
aerial vegetative unit; seedlings total weight;
tomato seedlings, especially in terms of plant
root weight and volume;
height (R2 = 0.573) (Figure 1).
measurements of the main physiological In order to determine the overall effect and
processes intensity (photosynthesis, influence of the treatments program applied on
transpiration, stomatal conductance) at the end the tomatoes seedlings were made observations
of the experiment. We used the LC pro+ and measurements also one week after
photosynthesis system. The measurements were application of the second treatment. The results
performed on the active leaves located in the obtained are shown in Tables 2 and 3,
middle third part of the plant. respectively in Figures 2, 3 and 4.
387
Table 2. Growth of tomatoes seedlings at one week after
the second treatment
Variant No. Plants Roots Plants Leaves
of height length total frequency
leaves HPA HR lenght (no./cm
(cm) (cm) HT(cm) HPA)
V1 6.2 34.6 21.2 55.8 0.18
V2 6.8 33.6 15.8 49.4 0.20
V3 8.4 32.4 18.4 50.8 0.26
V4 7.6 32.6 15.6 48.2 0.23
V5 8.2 32.4 16.4 48.8 0.25
Figure 3. Influence of applied treatment on the frequency
leaves of the tomatoes seedlings one week after the
Applied treatment program determined second treatment
differences regarding on the growth of
tomatoes seedlings. It is noteworthy that all Regarding on plants growth, taken together the
variants where have been applied different results obtained for tomatoes seedlings
treatments with bioactive substances have morphometry, we estimate that the most
achieved a shorter plant heights and roots balanced variant is V3 (fertilized with
lengths, compared to the untreated control Razormin 0.1%) followed by V5 (Bio Roots
variant. 0.2%).
Analysis of the results on the growth of Developing of tomatoes seedlings one week
seedlings showed that the best option working after the second treatments was quantified by
was V3-Razormin 0.1%. In this variant plants various indicators of mass and volume and by
have achieved the best and balanced growth, all diameter of collet. The obtained results (Table
indicators analyzed had very good values (8.4 3; Figures 4 and 5) regarding tomatoes mass
leaves formed, 32.4 cm plant height, 18.4 cm ratio highlights two situations:
root length, total length 50.8 cm plant; 0.26 1. At V3 variant all indicators recorded the
frequency leaves). In contrast, untreated V1 highest values compare to the other variants
seedlings, although recorded the highest plant (aerial part mass 16 g; mass root 4.5 g; 23.5 g
height (34.6 cm) and the largest length of the total mass, volume roots 8.5 cm3; 7 mm collet
roots (21.2 cm), achieved the lowest number of diameter). Good results have also recorded the
leaves (6.2 leaves) and respectively the lowest seedlings treated with Bio Roots and BAC
frequency of leaves, 0.18. This context, for V1, Foliar Spray.
shows an unbalanced growth that is based on 2. In contrast, V1 variant recorded, to all most
an elongation phenomenon due to the presence indicators, the lowest values (aerial part mass
of stress factors. 11.5 g; 19 g total mass; volume roots 2.5 cm3;
3.5 mm collet diameter), excluding the roots
mass (7.5 g) and roots volume (8 cm3).
Table 3. Developing of tomatoes seedlings at one week
after the second treatments
Aerial
Roots Total Roots
part Ø
mass mass volume
Variant mass collet
MR TM VR
MPA (mm)
(g) (g) (cm3)
(g)
Figure 2. Influence of applied treatment on tomatoes V1 11.5 7.5 19.0 8.0 5.8
seedlings growth one week after the second treatment V2 12.0 7.0 19.0 8.0 6.0
V3 16.0 7.5 23.5 8.5 7.0
As can be seen from Figures 2 and 3, schedule V4 15.5 6.0 21.5 6.5 7.0
treatments with bioactive substances exert a V5 15.8 7.5 23.3 8.5 7.0
distinctive influence on the general growth of
tomatoes seedlings, respectively, on the Also, was noticed a strong influence of the
frequency leaves (R²=0.639) and on the plant applied treatment on the aerial part mass and of
height (R² = 0.573). the collet diameter (Figure 4).
388
CONCLUSIONS
In the last few years, amid the development of

integrated and biologically horticultural
concepts, it has been necessary to reconsider
fertilizers used in horticultural practice by
introducing into the technological stream new
complex substances such as phytohormones or
biofertilizers, natural and non-polluting.
Figure 4 Influence of applied treatment on the aerial part
mass and collet diameter at the second moment
Treatments with bioactive substance are
applied with good results in vegetable practice
The results of the physiological measurements to accelerate or inhibit the growth of seedlings
performed on the experimental variants are or to support plants life in various stress
shown in Table 4. As can be seen, the leaf situations.
temperature was relatively constant (27.2- Tomato seedlings from the very first moment
28.3oC) and light intensity (Q) registered the of analysis showed important nuances as a
value of 1280-1360 mmol/m2/s. result of the application of bioactive substances
Table 4. Physiology of the tomatoes seedlings one week treatments in sense that, in the untreated V1
after the second treatment variant, the plants have recorded biometric
Var. A E A/E Leaf Q indicators far inferior to those of the treated
[µmol/ [µmol/ temp.T [μmoli/ variants and the context analysis highlights V4-
m2/s] m2/s] [0C] m2/s] fertilized with BAC Foliar Spray 0.3% as the
V1 11.35 3.11 3.65 28.3 1280
V2 12.13 1.60* 7.58 28.3 1280 seedlings had the most balanced growth and
V3 13.39 1.74 7.69 27.2 1360 development.
V4 12.39 1.87 6.62 27.2 1360 Analysis of the results of the growth and
V5 12.96 1.78 7.28 27.2 1360
development of tomatoes seedlings at the end
of experimental period showed that the best
The results analysis revealed that V3 recorded option working was V3-Razormin 0.1%. In this
the highest values of Photosynthesis rate A = variant plants have achieved the best and
13.39μmol/m2/s and efficiency (A/E = 7.69) on balanced growth and development, all
the backgroundof a low Transpiration rate E = indicators analyzed had very good values. This
1.74 μmol/m2/s. Also, V5 recorded high values is followed by V5-Bio Roots 0.2%
for the physiologic indicators. These results Physiologic activities revealed that V3 recorded
confirm the good growth and development of the highest values of Photosynthesis rate and
seedlings from these two variants. efficiency on the background of a low
V1 untreated recorded a very good Transpiration rate. Also, V5 recorded high
Photosynthesis rate (A = 11.35 μmol/m2/s) but values for the physiologic indicators.
very intense Transpiration rate (E = 3.11 These results confirm the good growth and
μmol/m2/s) determined a low efficiency of development of seedlings in the two mentioned
photosynthesis (A/E = 3.65). The intensify of variants (Razormin 0.1 % and Bio Roots 0.2 %)
physiological processes without translocation which we recommended as support treatment to
and accumulation of photoassimilated improving quality of tomatoes seedlings.
substances is, in fact, the response of plants to
the action of stressors whose intensity action REFERENCES
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overall results can be noted V4 (BAC Foliar Bălan D., Dobrin E., Luţă G., Gherghina E., 2018.
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390
INTER-SPECIFIC (CAPSICUM CHACOENSE HUNZ.

AND CAPSICUM ANNUUM L.) INHERITANCE OF FRUIT DETACHMENT
FORCE TRAIT IN HOT PEPPER
Bekir Bülent ARPACI, Faika YARALI KARAKAN
Kilis 7 Aralık University, Faculty of Agriculture, Department of Horticulture,

Mehmet Sanlı Mah. Doğan Güreş Paşa Bul. No:134, KİLİS, Turkey
Abstract
In red pepper cultivation for spice production, harvesting the fruits is time consuming and high cost requiring process.
Also all the cultivated pepper (Capsicum spp.) fruits for processing have to be separated from the pedicel after or
before drying. The pedicel is tightly attached with the calyx to the fruit pod in most pepper cultivar. For this reason fruit
detachment force (FDF) is an important trait to improve pepper cultivars suitable for mechanical harvesting. Gene
action for FDF was calculated using inter-specific crosses between four C. annuum L. (3.860 to 7.340 N FDF)
genotypes and C. chacoense Hunz. (0.199 N FDF), C. annuum variety ‘Totolapa’ (0.173 N FDF). FDF values obtained
from parents and F1 generation indicated that gene effects for this trait were mostly displayed dominance and additive.
No differences were calculated with respect to heterosis both Capsicum species related FDF. Parents could generate
hybrids with high degree of negative heterosis varying between 86.56% and 94.50% related with FDF. Except K7 x CC
hybrid easily detached from the fruit with 0.153 N, all the hybrids have fruit detachment force over ʻTotolapaʼ.
Key words: Fruit detachment force, heterosis, gene effect, pepper.
INTRODUCTION line × tester parental design using interspecific

crossing in pepper.
Chilies (dried red pepper) is one of the most
important crops in South East Anatolian MATERIALS AND METHODS
Region where produced 95% of 228,531 tons
total chili production of Turkey. Capsicum annuum (Totolapa) and Capsicum
Harvesting of chilies requires high labor and chacoense (C. cha.) exhibited easy fruit
cost due to pick the fruits by hand. Harvest detaching traits were used as male and
machines for dried chilies have been developed Capsicum annuum (Dila, Sena, K7 and K8)
but cultivars suitable for mechanically exhibited hard genotypes were used as female
harvesting needs some improvements (Palau parents. FDF was measured at red mature stage
and Torregrosa, 1996; Akay et al., 2009). using with two different interval (0.001-5 and
Fruit detachment force (FDF) is the most 0.1-500 N) digital push and pull force gauge as
important trait related with either hand or Newton (Geratech Inc. SH-5 and SH-50)
mechanically harvesting. (Figure 1). Variance of general and specific
Inheritance of detachment trait firstly reported combining ability effects and heterosis were
with single dominant gene (Smith, 1951) and calculated by TNAUSTAT-Statistical package
softening of fruit considered another gene (Manivannan, 2014) with line × tester mating
controlling to soft fruited trait. Gersch et al. design (Singh and Chaudhry, 1985) using
(1998) correlated FDF with cell size and cell testers as male (Totolapa and C. cha.) and lines
wall thickness in fruit and calyx. female (Dila, Sena, K7 and K8). Experiment
Finally Rao and Paran (2003) detachment or was designed completely randomized blocks
deciduous fruit and soft fruited traits were with 15 replicates. Mean differences among
revealed by pleiotropic effect of endo- hybrids and parents related with FDF were
polygalacturonase (PG) gene. With this article compared LS Means Differences Tukey HSD.
the gene action of FDF was calculated with Two hybrid population of Capsicum chacoense
391
and Capsicum annuum species matched by
paired sample t-test. JMP 5.0.1 software was
used for statistical analysis.
The fruit detachment trait which is wild

characteristic (Paran and Knapp, 2009; Mao
and Motsenbocker, 2001) decrease in
population with the domestication of pepper.
Fruits of many pepper cultivars using fresh or
processed tightly adhere to the calyx
(Motsenbocker, 1996). Due to time consuming
and great effort requirements both harvesting
and post-harvest process detachment trait come
into prominence for harvest technology so that
to eliminate the pedicel from the pepper fruit.
Table 1. Analysis of variance for Line × Tester and

Combining ability analysis for FDF traits
Line × Tester (Including parents)
SOURCE DF SS MSS F ratio
Replicates 14 2.40 0.1715 0.24
Genotypes 13 1443.06 111.0049 157.85
Cross 7 0.19 0.0278 0.03
Line (c) 3 0.09 0.0320 0.04 Figure 1.Measuring of FDF using push-pull digital gauge
Test (c) 1 0.04 0.0488 0.06
L x T (c) 3 0.04 0.0165 0.02 Variance of general combining (σ2GCA) and of
Parent 5 705.15 141.0307 200.55
Line (p) 3 98.82 32.9429 46.84
lines (σ2SCA) related crosses in respect to FDF
Test (p) 1 0.92 0.9289 1.32 were shown in Table 2. General combining
L (p) vt (p) 1 605.39 605.3961 860.89 ability of Dila and Sena varieties was found
CrovsPAR 1 737.71 737.7152 1049.05 non-significant on account of FDF while K7
Error 182 127.98 0.7032 and K8 has significant (p<0.01) general
Total 209 1573.44
Combining ability analysis
combining ability among the lines using as
Replicates 14 0.10 0.0075 2.01 female plants. GCA effects of both testers were
Cross 7 0.19 0.0278 7.43 significant for FDF trait. Variance of specific
Line (c) 3 0.09 0.0320 8.57 combining ability (σ2SCA) was non-significant
Test (c) 1 0.04 0.0488 13.07 for all of the crosses. Variance of general
L x T (c) 3 0.04 0.0165 4.42
combining ability was found greater than that
Error 98 0.36 0.0037
Total 119 0.66 of specific combination ability. This situation
Proportional contribution of lines, testers and their indicated that additive genes could be effect on
interactions to total variance FDF traits. Werner and Honma (1980)
Contribution of lines 49.39 explained fruit detachment at the fruit
Contribution of testers 25.10 receptacle controlling by additive gene effects
Contribution of L × T 25.51
and pointed out no differences between
reciprocal crosses. However mid, low, better
According to analysis of variance for line ×
parent and standard heterosis percentage of
tester analysis cross and lines significantly
hybrids for FDF were found significantly and
(p<0.05) important contribution of FDF traits.
negatively. The lowest FDF values are
Percentage of contribution related with FDF
important for easy pick by hand and harvest for
49.39, 25.10 and 25.51 for lines, testers and
the fruits (Werner and Honma, 1980;
lines × testers, respectively (Table 1).
Motsenbocker, 1996; Gersch et al., 1998).
392
Therefore negative heterosis is expected from along with soft fruit flesh dominantly.
the hybrids for this trait. The highest negative Nevertheless fruit detachment force associated
mid parent heterosis percentage was observed with deciduous trait was impressed on additive
K7 x Totolapa with -95.12%. Low parent gene effects combination with dominance. Rao
heterosis was -97.03% at Dila x C. cha. and and Paran (2003) determined deciduous fruit
better and standard heterosis percentage same and soft fruited trait under control pleiotropic
at this hybrid because of using Dila variety as effect of PG gene.
standard to calculate standard heterosis. But the This study exhibited that expression of PG gene
highest negative standard heterosis percentage could be diversified of phenotype. Gersch et al.
was obtained from K7 x Totolapa hybrid with- (1998) emphasized that FDF related with
97.82% (Table 2). All of the hybrids have genotype, maturity and plant growth regulator
deciduous fruit and soft fruit flesh. For this and also was affected by changing environment
reason deciduous fruit trait was expressed such as greenhouse and field.
Table 2. General and specific combining ability effects and heterosis percentage of lines testers and crosses for FDF
Heterosis (%)
Lines σ2GCA Cross σ2SCA Mid Parent Low Parent Better Parent Standard
Dila 0.01 ns Dila x C. cha. -0.02ns -94.50** -97.03** -97.03** -97.03**
Sena 0.01 ns Sena x C. cha. 0.02ns -92.12** -95.96** -95.96** -95.96**
K7 0.04** K7 x C. cha. 0.03ns -93.83** -96.69** -96.69** -96.39**
K8 -0.04** K8 x C. cha. -0.03ns -93.89** -96.87** -96.87** -96.58**
Testers Dila x Totolapa 0.01ns -88.32** -93.38** -93.38** -96.09**
Totolapa -0.02* Sena x Totolapa -0.01ns -86.56** -92.98** -92.98** -95.86**
C.chacoense 0.02* K7 x Totolapa -0.02ns -95.12** -97.33** -97.33** -97.82**
K8 x Totolapa 0.02ns -91.74** -95.74** -95.74** -96.52**
*
Significant differences at p<0.05 ** p<0.01 and ns: non-significant
Figure 2. Fruit Detachment Force (FDF) of lines tester and crosses.

x
Mean separation within the columns by LSMeans Differences Tukey HSD multiple range test at p≤ 0.05
Fruit detachment force of lines, testers and testers and crosses. However all of the crosses
and testers placed same statistically group
crosses were demonstrated in Figure 2. The
(Figure 2.). Motsenbocker (1996) determined
highest FDF was 7.340 N at Sena variety. K7
was the less adhere to the receptacle genotypes the lowest FDF value with 2.5 N at red mature
in the lines with 3.860 N FDF. The testers stage “McIlhenny Select” tabasco (Capsicum
Totolapa and C. cha. displayed 0.173 and 0.185 frutescens) pepper variety in field study while it
N FDF, respectively. K7 x Totolapa hybrid was 0.6 N in greenhouse condition. Mao and
exhibited the lowest FDF among all the lines, Motsenbocker (2002) observed 2.9 N FDF
393
from McIlhenny Select and no differences find Research of Institute with the assistance of
between ethephon applied and control Ayhan AK in Kahramanmaraş. We thank to
treatment on account of FDF. With this study stuffs of this institute for their assistance.
two F1 populations of Capsicum chacoense and
Capsicum annuum species were compared. No REFERENCES
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High percentage of mid, low, better parent and Ra, G.U., Paran I., 2003. Polygalacturonase: a candidate
standard heterosis was calculated for FDF gene for the soft flesh and deciduous fruit mutation in
traits. Quite low FDF was observed on crosses Capsicum. Plant Molecular Biology, 51 (1), 135-141.
from two different Capsicum species. These Singh R.K., Chaudhary B.D., 1985. Line × Tester
analysis. Biometrical Methods in Quantitative
FDF values are promising to improve pepper Genetic Analysis, Ed, 3, 215-223.
varieties for mechanical harvesting. Smith P.G., 1951. Deciduous ripe fruit character in
peppers. Proc. Amer. Soc. Hort. Sci. 57: 343-344.
ACKNOWLEDGEMENTS Werner D.J., Honma S., 1980. Inheritance of fruit
detachment force in pepper, Journal of the American
Society for Horticultural Science 105.6 (1980): 805-
This research work was carried out in East 807.
Mediterranean Transitional Zone Agricultural
394
ASSESSING TOMATO GERMPLASM FROM V.R.D.S. BUZAU
TO IDENTIFY GENOTYPES WITH DISTINCT FEATURES
Bianca ZAMFIR1, Dorel HOZA2, Costel VÎNĂTORU1, Camelia BRATU1, Elena BARCANU1
1
Vegetable Research and Development Station Buzău, No. 23, Mesteacănului Street,
zip code 120024, Buzău, Romania
2

Abstract
V.R.D.S. Buzau is well known for tomato breeding, over time here were obtained valuable varieties appreciated by both
consumers and growers. Currently, V.R.D.S. Buzau has an important germplasm collection consisting of over 1500
genotypes in different breeding phases. Since 1996, research on breeding this species were intensively undertaken, been
obtained for the first time in our country hybrids with a certain destination. The research carried out so far has mainly
focused on the germplasm evaluation accumulated to identify genotypes with distinct features for the breeding process,
as well as availability and genetic stability in the progeny. The study aimed the observation of shape, size and fruit
color to identify distinct features. The stable identified genotypes were organized into two groups: cherry type varieties
and large fruits of different shapes like pear, bell peppers, long pepper, lemon, banana varieties, with different colors
like white, various shades of yellow, red, pink, brown, black, burgundy varieties. Two of these genotypes have been
proposed for patenting and are to be expanded on a large scale in production.
Key words: biodiversity, breeding, Estera, Hera, specific destination.
INTRODUCTION applications in both science and commerce.

Current efforts in tomato breeding are focused
Tomatoes were brought to Europe quite late, in on discovering and exploiting genes for the
the 16th century and more lately in Romania, most important traits in tomato germplasm. In
19th century. Although they were brought the future, breeders will design cultivars by a
recently in Romania, this species is the most process named ‘breeding by design’ based on
cultivated among vegetables. Edible tomato has the combination of science and technologies
its origin in the small and yellow wild tomato. from the genomic era as well as their practical
Both plant and fruits sepals of the wild tomato skills (Yuling Bai, 2007). Beside the recent
are covered by big aggressive spines. Cultivars breeding aims, crop yield and quality remain
with red edible fruits of different sizes were the main objectives The major goals of tomato
obtained by selection and breeding methods breeders (higher productivity, better tolerance
and nowadays are cultivated on large scale. to biotic and abiotic stresses and increased
Domestication is often controlled by a limited sensory and health value of the fruit) require a
number of chromosomal regions with major good comprehension and management of
phenotypic effect In tomato, edible fruits, tomato genetic resources diversity (Guillaume
attractive red colour and fruit size increase are Bauchet, 2012). V.R.D.S. Buzau was interested
characterizing this process (Purugganan et al, in breeding this species since its establishment
2009). in 1957. There were obtained well known and
Following the selection and modern breeding appreciated varieties by both consumers and
methods, nowadays were obtained numerous growers. Since 1996 the research was focused
genotypes of this species with distinct on enriching the Official Crop Plants List of
phenotypic traits. Also the consumers and Romania with new tomato varieties.
producers demands imposed new breeding
directions to obtain cultivars with precise use. MATERIALS AND METHODS
From first domestication to modern breeding,
the tomato has been continually subjected to Research started with collecting and assessing
human selection for a wide array of the tomato germplasm and breeding the
395
valuable genotypes. Now the germplasm Concerning the plant height, A 80 reached the
collection contains over 1500 genotypes in highest value of 291 cm. A 631 has the richest
different breeding phases. The genotypes foliage, this feature protects the fruits against
assessing and organizing was made on the solar burns.
following criteria: type of growth and progeny Maximum number of trusses per plant was
genetic stability (Figure 1). scored by A 709 with 11 trusses.
Concerning the fruits features, there were
2000 selected accessions with distinct traits which
demonstrates the diversity of this species.
1500 Thereby A 631 presents yellow lemon shaped
fruits, productive plant and the fruit weights
1000 83.3 g on the average. A 312 has cherry striped
fruits, dark brown with red stripes and dark red
500 pulp. The fruit weights 26.3 g on the average
and has good firmness. A 524 has bell pepper
0
shaped fruits, red coloured at maturity that
Total SP+ S A Sg SP S A Sg Sp S A Sg
weights 233.6 g. The fruits have large interior
Figure 1. Tomato germplasm grouped on: spaces and can be used to prepare some dishes
SP+ (indeterminate) - 751 from which: S (Stable) 236, like stuffed peppers.
A (advanced) - 177, Sg (segregant) - 338; SP (semi-
determinate) - 374 from which: S 105, A 118,
A 522 has light yellow big fruits with red pulp,
Sg-151; Sp-(determinate) - 447 from which: S 200, with 252.3 g average weight. A 308 has red
A - 137 and Sg 110 with yellow stripes bell pepper fruit shaped and
very productive plants. A 306 has very tasty
Due to the large number of collected and cherry black fruits that weights 18.4 g on the
assessed genotypes, new selection and breeding average. A 532 has banana intense yellow
methods were implemented. shaped fruits that lengths 7.2 cm and A 709 is
Our study was focused on individual repeated the most tasty accession due to sugar content of
selection. Beside this, there were used 12-16%. A 724 is a different accession with
hybridization, segregation, mutations and dark black fruits and uniparous raceme.
controlled genetic drift. The pulp is dark red coloured. A 2000 has low
productive plants but very big tasty heart
RESULTS AND DISCUSSIONS shaped fruits. This accession comes from the
Ox heart heirloom (Table 2) (Figure 2)
Following the germplasm assessing there were From these accessions, A 80, A 28 and A 2000
retained 12 accessions that correspond to the were selected as the most interesting and
main breeding proposed objectives (Table 1). genetic stable genotypes that correspond to the
Concerning the main plant features, accession consumers and growers requirements.
(A) 532 has semi-determinate type of growth These accessions were registered for patenting
and the rest of the accessions have indeter- since 2016 at ISTIS Bucharest.
minate growth.
Table 1. The main plant features
Studied feature
631 312 524 522 28 308 306 532 709 80 724 2000
Accesion
Growth type SP+ SP+ SP+ SP+ SP+ SP+ SP+ SP SP+ SP+ SP+ SP+
Plant height (cm) 271 281 268 272 263 288 268 165 258 291 263 250
Lateral shoots 12 13 11 13 13 10 11 9 12 14 13 18
Leaves/plant 36 30 27 21 23 31 26 17 32 26 28 35
Leaf length (cm) 36 35 52 38 40 42 40 33 32 43 36 49
Trusses/plant 9 8 6 5 5 5 8 5 11 10 7 5
Distance between
22 24 34 27 28 31 36 28 35 20 21 28
trusses (cm)
396
Table 2. The main fruit features
The main Fruit weight Fruit height Fruit diameter Seminal Pulp thickness
Main distinct feature Sugar content (%)
studied feature (g) (cm) (cm) lodges (mm)
A 631 83,3 6,7 4,6 3 50 Lemon shaped fruit 5.3
A 312 26.3 3.7 3.2 2 50 Cherry striped fruit 11.8
A 524 233.6 8.9 7.4 4 70 Bell pepper fruit 6
A 522 252.3 5.5 9 16 50 Pineapple coloured fruit 4.3
A 28 111.9 12 3.5 2 50 Long pepper shaped 5.7
A 308 178.9 5.5 8.5 5 90 Bell pepper striped fruit 3.8
A 306 18.4 2.5 2.5 3 30 Black cherry fruit 7
A 532 75.6 7.2 3.9 3 50 Banana shaped fruit 7
A 709 11,1 4 2 2 30 Pear shaped fruit 12-16
A 80 12.1 3.6 2.1 2 40 Ovoid cherry fruit 11.2
A 724 42,8 4,7 3,9 3 70 Black fruits 5.8
heart shaped big tasty
A 2000 560 9,5 10,3 20 70 5.4
fruits
The accession has big fruits, long pepper

shaped ranged in 100-150 g weight with high
content of dry matter.
The main feature of this accession is the shape
A 522 A 308 A 709
of the fruit. These are big in length slightly
pointed (Figure 4).
A 631 A 524 A 724

Figure 2. Studied accesions
Accession 80 presents indeterminate growth

and can be cultivated both for fresh
consumption and processing, can be cultivated Figure 4. Crop detail of A 28
both in open field and greenhouse, in trailing
system. The accession has ovoid fruits that A 2000 has indeterminate growth and can be
weights 16-20 g and a reduced number of cultivated in trailing system, in open field.
seeds, < 20 seeds/fruit. The fruits have a high The genetic material comes from the Ox heart
content of sugar, 11.2%. heirloom.
The main feature of this accession is its distinct This accession has big fruits that ranged 300-
taste and flavor. The fruits tastings showed that 950 g weight/fruit, heart shaped with green
this accession is superior in what concerns taste shoulder at the physiological maturity.
quality in comparison with the other accessions The accession is different due to specific taste
from the germplasm collection (Figure 3). and flavour of traditional Romanian tomato
A 28 has indeterminate growth and can be (Figure 5).
cultivated both in open field and greenhouses,
in trailing system, both for fresh consumption
and for processing.
Figure 5. Crop detail of A 2000
Figure 3. Crop detail of A 80

397
The research undertaken until now ended with Guillaume B., Causse M., 2012. Genetic Diversity in
collection and assessing the germplasm Tomato (Solanum lycopersicum) and Its Wild
Relatives, Genetic Diversity in Plants, Prof. Mahmut
collection and organizing it according to Caliskan (Ed.), ISBN: 978-953-51-0185-7, In Tech,
genetic stability and type of growth. http://www.intechopen.com/books/genetic-diversity-
We obtained 12 genotypes with distinct in-plants/genetic-diversity-in-tomatosolanum-
features which correspond to the main breeding lycopersicum-and-its-wild-relatives.
aims. Purugganan M.D., Fuller D.Q., 2009, The nature of
selection during plant domestication. Nature 457
A 80 patented under the name of Estera, A 28 (7231): 843-848.
patented as Hera and A 2000, Bizon, were Yuling Bai, Pim Lindhout, 2007. Domestication and
registered for approval and patenting. Breeding of Tomatoes: What have We Gained and
What Can We Gain in the Future? Annals of Botany,
Volume 100, Issue 5, no. 1, 1085-1094,
https://doi.org/10.1093/aob/mcm150.
398
NEW EGGPLANT GENOTYPES WITH DISTINCT PHENOTYPIC

EXPRESSIVITY OBTAINED AT V.R.D.S BUZĂU
Camelia BRATU1, Florin STĂNICĂ2, Costel VÎNĂTORU1, Bianca ZAMFIR1,

Elena BARCANU1
1
Vegetable Research and Development Station Buzău, No. 23, Mesteacănului Street,
zip code 120024, Buzău, Romania
2
Abstract
V.R.D.S. Buzău has a great tradition in the process of breeding for this species. Here were maintained by conservative
selection the first valuable Romanian eggplants like ʻDanubianaʼ, ʻBucuresteneʼ, ʻPana Corbuluiʼ along with ʻZarazaʼ
and ʻDragaicaʼ. Recently, the first hybrid of eggplants obtained by the Breeding Laboratory was patented, and
registered in the Romanian Official Catalogue for Crop Plants under the name of ʻRebeca F1ʼ. As a result of the
researches made over time the Laboratory gathered an important germplasm collection and numerous informations
regarding the activity of breeding this species. At this time, the germplasm collection is being compound of over 60
valuable genotypes with distinct phenotypic expressivity for their shape, size and colour. Stable genotypes that are a
very important part of this paper were obtained. These have large white fruits, red coloured, brindled, green etc. Of
these, the accession A 10 (Camelia), with large white fruits is in the final testing stage for patenting and will be
extended on a large scale in production.
Key words: aubergine, breeding, Camelia, germplasm collection, Solanum melongena.
INTRODUCTION creation was patented and registered in the

Official Catalogue for Romanian Crop Plants
„The eggplant arrived in Europe around 1300, under the name of ‘Rebeca F1’. If in the past
and the eggplant fruits were used as food after the requirements of processors and consumers
the sixteenth century” (Cakir et al., 2017). were limited at cultivars with traditional fruits,
In Romania, the eggplant crop production was black or violet, of ovoid or cylindrical shape,
introduced after the 19th century. In the early now, their requirements have diversified a lot.
times, foreign varieties were used in order to ‘Vegetables from imports are more varied than
establish the crops, and the mainly varieties those derived from domestic production (Soare
used for this purpose were: ‘Delicates’, et al., 2016).
‘Odesa’, ‘Lungi violete’. The first Romanian Therefore since 1996, in establishing the
creations in this species were achieved after the breeding objectives the buyer’s actual require-
Research Stations were founded, and the ments were taken into consideration. Modern
earliest and most used were ‘Danubiana’, breeding methods and a varied genetic material
‘Bucurestene’ and ‘Pana Corbului’. VRDS were implemented allowing obtaining a great
Buzau gave a special attention for breeding number of genotypes with distinct phenotypic
eggplants since April 1957, this being the year characteristics included in this paperwork.
of it’s founding, and managed to create two
very valuable varieties that are still being used MATERIALS AND METHODS
and appreciated: ‘Dragaica’ and ‘Zaraza’.
„Now there are an increasing number of Researches debuted by achieving the germ-
F1hybrid varieties bred by seed companies and plasm collection. The main focus was on
the seed production of egg-plant is shifting valorisation the autochthonous genetic potential
from farmer’s hands to seed companies” (Chen consisting of varieties and local populations.
et al., 2001). Recently, the first eggplant hybrid The majority of the genotypes within the
399
collection belong to the Solanum melongena
species, but among these are also included va-
rieties from S. macrocarpon and S. ethiopicum.
Therefore, the total collection for the eggplant
complex collection is of 286 accessions.
After establishing the eggplant collection,
researches were continued by evaluating the
genetic stability in lineage and by identifying
the useful characteristics for the breeding A10 A20 A21B
process.
The main breeding methods used were inter
and intraspecific hybridization, the segregating
phenomenon and also by breeding the valuable
genotypes thru repeated individual selection.
The main breeding objectives were channelled
towards productivity, earliness and phenotypic A22A A23A A26
distinctiveness.
A special emphasis was placed on breeding the
lines that present a special colour variation
from white, purple, green, with stripes, patches,
and in various shape and size, small, cylin-
drical, globular, ovate, pear shaped, club
shaped, ellipsoid, cylindrical, ribbed.
Simultaneously, genetic stability was A29A A30C
monitored. Researches were made in protected
areas (greenhouses without a heating system).
Sowing was made in alveolar trays with 70
orifices, using as a substrate peat, in the first
decade of March.
The seedlings were planted in the second
decade of April, using the distance for planting
of 70 cm between rows and 40 cm between A51A A58
plants per row. It was kept a distance of 120 cm
Figure 1. Aspects of the selected genotypes
between the planting strips.
The cultivation technology applied was the one A23A has greenish rounded and ribbed fruits
specific to this species, and the special works that are small in size and have a good store
consisted of trellising and pruning. The selected resistance.
accessions are: A10, A20, A21B, A22A, A23A, A26 has medium large fruits, round and a very
A26, A29A, A30C, A51A, A58 (Figure 1). pleasant appearance due to its purple skin
A10 presents the greatest interest for the coloration.
present work has large white fruits, very A29A was selected for the plant vigour and
productive with a very pleasant commercial green fruits that when are fully ripen turn
appearance. orange-red, their appearance being very similar
A20 has very small fruits, similar in shape and to a small pumpkin.
size with an egg, this being the most attractive A30C has white cylindrical small fruits that
feature of it. Also it has a great number of produce a great number of fruits with a good
fruits, over 40 allowing the harvesting for the storage capacity.
entire cycle of production. A51A has typical purple ellipsoid fruits. It is a
A21B has white fruits with purple stripes. very productive accession with a good
These are of a medium size with a good yield resistance to pests and diseases. The fruits are
production. weighing between 700 g and 1100 g, depending
on the technology used.
400
A58 has very interesting fruits due to their side A26 has the smallest height of 99 cm-
green coloration and club shaped appearance. mean value.
Stem length varies from 6 cm to 19 cm with no
RESULTS AND DISCUSSIONS correlation between plant height and stem
length. A10 has a medium height value of 162
Researches started with the achievement of a cm but is very well developed due to the
rich germplasm collection followed by its number of side shoots (3) which gives it an
evaluation and acknowledgement regarding its aspect of fullness. These accessions present
genetic stability and also inventing the useful different leaf sizes and shapes correlated with
characters for the breeding process. the plant vigour.
After establishing and evaluating the
germplasm collection, this was structured on 3 Table2. Fruit characteristics-mean values
groups according to their genetic stability. Of
Plant
286 accessions studied, a number of 62 features
A10 A20 A21B A22A A23A
accessions showed genetic stability in lineage, Genotype
Peduncle and sepals
86 accessions were included in the genetically colour
Green Green Green Green Green
advanced genotypes group these being the
Peduncle length (cm) 9 3.08 5.1 4.4 3.82
genotypes that proved a small variation
On Absen On Absen Absen
regarding the variation expressiveness of the Presence of spines
sepals t sepals t t
main characteristics. White White
Fruit colour at lined lined
A number of 138 genotypes form the group of consumption maturity
White White
with
White
with
segregating genotypes. purple green
These ones present a great variability regarding Fruit colour at
Yello
w
the expressiveness of the main characters, the physiological
Yello
w
Yello
w
lined
Yello
w
Yello
w
majority originating from the segregation of maturity with
brown
hybrid combinations made during the breeding Fruit length (cm) 18 5.1 11 13 4.83
program. Median fruit diameter
9.5 3.71 7 6.2 6.30
(cm)
From the group of stable genotypes 10 acces- Plant
sions with distinct phenotypic expressivity features
A26 A29A A30C A51A A58
Genotype
regarding colour, shape and fruit size were Peduncle and sepals
Purple Green Green Green Green
selected. The main plant characteristics are colour
Peduncle length (cm) 3.9 1.74 4.52 9 4.5
shown in table 1. On Absen On On On
Presence of spines
sepals t sepals sepals sepals
Table1. Main plant characteristics-mean values Fruit colour at
Purple Green White Purple Green
consumption maturity
Plant Fruit colour at
Brow Yello Brow Yello
features physiological Red
nish w nish w
A10 A20 A21B A22A A23A maturity
Genotype
Fruit length (cm) 9.2 3.25 11.13 22 23
Plant height (cm) 162 155 200 100 156 Median fruit diameter
Stem length(cm) 8 8 13 6 19 7.8 5.10 3.69 12.1 4.05
(cm)
No. of side shoots 3 3 2 3 3
Leaf length(cm) 24 15 25 18 22.5
Leaf width(cm) 17 10 15 11 13 According to the data registered in Table 2,
Plant accession A10 differs from the other white
features
A26 A29A A30C A51A A58 eggplants presented, through the size of the
Genotype
fruit, having larger fruit than the others, as we
Plant height (cm) 99 140 114 165 110
Stem length(cm) 17 12 11 14 16
can see accession A10 has 18 cm fruit length
No. of side shoots 2 2 3 2 2 and the others have fruits of 13 cm and even
Leaf length(cm)
Leaf width(cm)
16
7
21
13
26
14
24
18
25.5
14
5.1 cm. Also the proportion between fruit
length and diameter (18 cm/9.5 cm) suggests
As shown in table no. 1, the accessions studied the good commercial ellipsoid aspect that
present different heights, A21B being the one makes it very attractive for the production
with the most luxurious growth, reaching in market.
mean values 200 cm length, and on the other
401
Table 3. Fruit harvest-mean values
Plant
features
A10 A20 A21B A22A A23A A26 A29A A30C A51A A58
Genotype
Fruit weight (g) 615 31 420 290 125 220.4 32 94 880 164
No. of fruits/plant 11 42 13 19 27 22 45 26 9 21
STAS I production (g) 4305 985 4120 3920 2896 3274 1115 2096 5842 3094
STAS II production (g) 1830 264 986 1260 345 964 284 279 1111 283
Substandard 630 53 354 330 134 610.8 41 69 967 67
Total production /plant (g) 6765 1302 5460 5510 3375 4848.8 1440 2444 7920 3444
As shown in Table 3, the most productive its evaluation according their genetic stability
accession is A51A that presents typical large and directions of use.
purple-black fruits followed with a difference Ten new genotypes were identified with
of 1155g by A10, which is our point of interest pronounced traits of distinctiveness especially
with a production of 6765 g/plant. For A10, the in matter of shape, size and fruit colour. Of
substandard fruit production represents 9.313% these, A10 accession was registered to ISTIS
of total fruit production/plant, while the STAS I Bucharest for patenting under the name of
production is 63.637% and STAS II is 27.051% Camelia. In the present it is in the 2nd year for
of fruit production/plant. The lowest production further testing, and the feedback collected from
was registered at A20, with the specification the partner farmers from the main vegetable
that this genotype has small fruits that are very holdings are positive. This new achieved
attractive for culinary uses, being a great genotype has large white fruits, with few seeds
replacement for mushrooms. in the fruit and a pleasant commercial
After a further evaluation regarding the com- appearance. It has a specific flavour and taste, a
mercial quality of the fruits the first accession buttery pulp with the specification that the
is A51A with a medium STAS I production of organoleptic qualities don’t alter during
fruits/plant of 5842 g, followed by A10 with a processing. It can be cultivated both in
medium STAS I production of fruits/plant of protected areas and in open field in all the areas
4305. The harvest with the commercial aspect favourable for this culture. The research has
that was slightly depreciated was directed to been completed with the rehabilitation and
STAS II; the fruit were slightly deformed or reduction of the main character variables in
were having an extended blossom point. cultivar 51 A, a valuable local bio-creation that
All the small fruits, highly depreciated that has been neglected in culture for a long period
were not proper for commercial purposes were of time. Also for this new variety the
directed to the substandard group. We need to documentation for patenting was prepared and
specify that there were no interventions on the sent to ISTIS Bucharest. The identified and
technological flow with fertilizers or additional studied cultivars open new directions for use in
technological works. For the future researches culinary preparations.
will continue with the implementation of
modern fertilizing plans and pollen stimulation REFERENCES
factors.
Cakir Z. et al., 2017. The Morphological Diversity and
Applying these new factors, certainly the Fruit Characterization of Turkish Eggplant
productivity and quality of the harvest will (Solanumm elongena L.) Populations.Ekin J, 3.2: 34-
increase. In the applied experiences, the 44.
classical culture technology for eggplants was Chen N.C., 2001. Eggplant seed production. AVRDC
used in order to evaluate the real genetic International Cooperators’ Guide. Asian Vegetable
Research and Development Center, Shanhua, Taiwan,
potential. 1-14.
Soare E., Dobre I., David L., 2016. Economic Analysis
CONCLUSIONS In Vegetable Sector Of Romania. Economic
Analysis, 16.4.
Researches finalized with the establishment of Lagunovschi-Luchian V., Vinatoru C., 2016.
Legumicultura, Editura Alpha MDN, 2016, 289-295.
a valuable germplasm collection, followed by
402
STUDY OF THE EFFECT OF ULTRASOUND

ON VEGETABLE CROPS IN DIFFERENT EXPOSURES
Vangel CHILINGIROV, Manol DALLEV, Nikolina SHOPOVA, Irena DAKOVA,

Galya HRISTOVA, Ivan MITKOV
Agricultural University of Plovdiv, 12 Mendeleev Blvd, 4000, Plovdiv, Bulgaria
Abstract
The paper examines the impact of ultrasound on seeds of vegetable crops of the Apiaceae family in different exposition.
The results of these studies show that seed treatment with ultrasound influences seed quality sowing. Ultrasonic
treatment of the appropriate duration increases not only germination energy but also laboratory germination. It should
be borne in mind that ultrasound stimulation with a certain duration, depending on the culture, may also have a
negative effect.
Key words: ultrasound, vegetable crops, quality sowing.
INTRODUCTION the production of celery is considerably more

labor intensive than other root crops (Cholakov,
Vegetable production is one of the main sub- 2009). The areas occupied with celery in 2014
sectors of agriculture and an important share of are 5 ha and the production is 57 t.
plant growing. Root vegetables - carrot, parsley The purpose of this study is to investigate the
and celery are crops of great economic possibility of pre-treatment of seeds with
importance due to their high nutritional and ultrasound to increase germination and seed
biological value. germination. Carrot, parsley and celery are
Carrots (Daucus carota), which are the main two-year-old vegetable crops of the Apiaceae
root crop, are used both for fresh and canned family. Carrot refers to the genus Daucus,
consumption. Besides food, carrots are used in parsley to the genus Petroselinum, and celery
pharmacy and perfumery industry. For to the genus Apium.
rhythmic supply to the market and canning Various methods of pre-seed treatment have
industry, carrots can be sown from early spring been developed, such as pre-soaking the seeds
to early summer and early winter. Traditional in water, rooting, heat treatment, impact with
sowing times are the end of February - early physical agents, etc., aiming at improving their
March, the first half of June and the first half of seed qualities (Esfandiar F. et al., 2012).
December. In the system of organic farming, the use of
Parsley and celery are used as a spice and are certain physical factors on the biological
important for improving the palatability and development of crops is a modern method of
digestibility of the food. This is due to the achieving high yields. The use of physical
essential oils contained in them. With the factors is preferable to chemical treatments that
content of nutrients and especially of essential can lead to plant contamination (Kwiatkowska
oils, celery takes one of the first places among B. et al., 2011).
vegetable cultures used as seasonings. With the The positive influence of ultrasound on seed
content of nutrients and especially of essential sowing properties has been found in other crops
oils, celery takes one of the first places among such as rice (Liu Y. et al., 2003), wheat
vegetable cultures used as seasonings. (Yaldagard M. et al., 2007), dill (Esfandiar
Celery is the only vegetable crop that grows Fateh, 2012; Zhao Yan et al., 2012), sunflower
through seedlings. Due to its long vegetation (Machikowa et al., 2013) and others.
period, the nursery cultivation method and the The first scientific studies on the influence of
large requirements for agrotechnical factors, ultrasound on plant development date back to
403
the 30-40 years of the last century. Experiments For the control untreated seeds of the crops set
conducted in the Soviet Union show that to germinate in Petri dishes. Depending on the
ultrasonic oscillations have a beneficial effect duration of seed treatment, the following
on germination and subsequent plant options were tested:
development (Istomina O., Ostrovskiy E., Carrot:
1936; Davidov G.K., 1940). 1. Control;
The use of ultrasound to accelerate germination 2. Exposure 3 min.;
of seeds is effective only when they are placed 3. Exposure 6 min.;
in a liquid medium, since this method of 4. Exposure 12 min.;
treatment allows the absorption of water from Parsley:
the seed that causes germination (Gordon, 1. Control;
1963; Yaldagard et al., 2008). 2. Exposure 3 min.;
3. Exposure 6 min.;
MATERIALS AND METHODS 4. Exposure 12 min.;
Celery:
The survey was conducted in the period 2016- 1. Control;
2017 years in the Department of Mechanization 2. Exposure 3 min.;
and Department of Horticulture at the 3. Exposure 6 min.;
University of Plovdiv. 4. Exposure 12 min.;
The subject of the study was the carrot, parsley The experiment was set in 3 replicates and the
and celery seeds. An ultrasonic bath is used to number of seeded counts in each iteration was
perform the study (Figure 1). 100. After the treatment, the seeds were placed
in a thermostat at 24°C, with the number of
seed germinated daily. The counting continued
until the 14th day for carrot, 21 for celery and
28 for parsley.
Two parameters were determined to determine
the viability of seeds and their germination,
namely laboratory germination (%) and
germination (%). As a result of day-to-day
readings, the speed and cooperativity of seed
germination were determined.
Laboratory germination, showing normally
germinated pure seeds under optimum
conditions for a specified period of time, is
Figure 1. BRAND CT-405 determined by counting the number of
germinated seeds according to the accepted
The bathtub uses the principle of intense sound BDS standards (601 84) as follows:
pressure, created by an ultrasound source, was 1) On the 14th day for carrot;
born in a limited volume of liquid - a plurality 2) On the 28th day for parsley;
of microscopic bubbles are formed that shrink 3) On the 21st day for celery.
and expand in sync with the frequency at which The germinating energy, expressing the
they hit the surface of the objects placed in the percentage of normally germinated seeds under
liquid. optimum conditions, but for a shorter period of
Three exposures of ultrasound treatment were time, was reported to:
tested at 3, 6 and 12 minutes at a frequency of 1) 7th day for carrots (under BDS);
43-45 kHz. To perform the ultrasound 2) 10th for parsley (under BDS);
treatment, the seeds were placed in distilled 3) 10th for celery (under USSR).
water.
404
1. Germination energy (%)

Research results show that seed treatment with
ultrasound has a positive effect on the
germination energy indicator and is best shown
in carrots.
From the results presented in Figure 2, it is
seen that as the exposure increases, the
percentage of germination energy also
increases. Depending on the duration of the
ultrasound seed treatment, the germination Figure 3. Germination energy (%) of parsley seeds
energy increased from 30% as measured by the averaged over the period 2016-2017
control to 43% in the 3 minutes version.
Prolonged treatment has a stimulating effect
since the value of the test item increases
regularly and reaches 65% in the 6 minutes
variant and up to 100% in the 12 minutes
exposure. The excess over the control was
116.7% and 233.3%, respectively.
Under the influence of ultrasound stimulation,
the terminable energy recorded in parsley
(Figure 3) increased, although slightly, from
20% (control readings) to 26% in the 6 minutes
exposure scenario, and increased to 28% in the
Figure 4. Germination energy (%) of celery seeds,
exposure duration variant 3 min. The excess averaged over the period 2016-2017
over the control in these variants is 30% and
40%, respectively. 2. Cultivation (%)
Germination is a very important indicator of a
biological aspect that provides information on
the viability of the seeds, their suitability and
the determination of the seed norm (Murtazov,
1984). And with this main indicator, the trend
established in carrots to enhance the effect of
seed treatment with increasing duration is
retained (Figure 5).
Figure 2. Germinating energy (%) of carrot seed,

average for the period 2016-2017
Treatment of 12 minutes ultrasound parsley

seeds has a rather negative effect as no
germinating seed is reported at this exposure,
and germinating energy is 0%, respectively.
The treatment of celery seeds with ultrasound
(Figure 4) affects the germination energy only Figure 5. Laboratory germination (%) of carrot seed,
when its duration is 12 minutes. The reported averaged over the period 2016-2017
sparkling energy in this variant is 14%. For all
other variants, including the control, the One hundred percent germination was reported
germinating energy reading is 0%. in the 12 minutes exposure scenario, and it
405
should be noted that these values were recorded In Figure 7 shows the results for laboratory
as early as the 7th day when germinating energy germination of parsley seeds. The highest
was determined. The excess over the control percentage of laboratory germination was
for this variant is 58.7%. reported in a 6 min. exposition, with a 33.3%
In the two other exposures, the reported values increase over the control.
were 75% for variant 2 (3 minutes) at 19% over
the control and 82% germination in variant 3 (6
minutes) with the control over 30.2%. The
germination rate at the control was 63% lower
than the other variants.
Attention is paid to the results presented in
Figure 6 providing information on the speed
and cope of seed germination in carrots.
Figure 7. Laboratory germination (%) of parsley seeds

averaged over the period 2016-2017
Laboratory germination reported at the control

was 60%. For the other two exposures, the
reported test value is lower than that of the
control. While in the 3 minutes variant the
Figure 6. Germination of carrot seeds by day difference is only 7% in favour of the control,
in the 12 minutes seed treatment variant, the
It is seen that under the influence of ultrasound difference in untreated seed (control) is drastic
treatment with a duration of 6 and 12 minutes and reaches 40%.
seed germination begins on the 4th day after the Obviously, this indicator can be said to be
treatment, which is a day earlier than the negatively affected by the prolonged treatment
control. of the seeds with ultrasound - 12 min.
Attention is paid to the results reported in the The results presented in Figure 8 are quite
12 minutes variant, in which seed germination different, so it is difficult to establish any
is most common. Within 3 consecutive days, tendency or regularity. However, seed germina-
the number of germinated seeds in this tion may vary from day 7 to day 11, with the 3
variation reaches 100. minutes exposure being at its earliest, and at 12
By comparison, the control for the same period minutes exposure at the latest. The control
of time is the number of sprouts 30. In the 6 sprout starts on the 9th and ends on the 14th. For
minutes variant, seed germination ends on day all other variations, this process ends until the
9, and for the other two options, including the 19th day.
control - on the 11th day.
406
Figure 8. Sprouting seeds of parsley by day for the whole period.
In Figure 9 the results for laboratory

germination of celery are presented. Laboratory
germination was only reported for 6 minutes
and 12 minutes exposures. Under the influence
of 6 minutes treatment seed germination
reaches 60% at 0% for the control.
Significantly lower and at the same time higher
than that of the control is laboratory
germination in the variant with 12 min.
duration - 20%.
Figure 10. Germination of celery seeds by day

for the whole period
CONCLUSIONS
Seed treatment with ultrasound influences seed

sowing.
The optimum duration of ultrasound exposure
in carrots lasts for 12 minutes, and for parsley
and celery - 6 minutes.
Figure 9. Laboratory germination (%) of celery seeds, This method of seed treatment increases the
average for the period 2016-2017 carrots energy of carrots by up to 233.3% and
of parsley by up to 40%. Celery has 14%
The germination process begins at day 9 in the germinated energy at 0% for the control.
variant (Figure 9) with 6 minutes exposure and Ultrasonic treatment of the appropriate duration
at day 12 in variant 4 (12 minutes). In the same increases not only germinating energy but also
two variants seed germination ends on day 17 laboratory germination. In carrots it reaches
on day one and on day 18 in the second 100%, in parsley 80%, and in celery - up to
variation. There is also a difference between 60%. The excess over the control with 58.7%
the variants in the seed germination rate. While for carrots and 33.3% for parsley. In celery
in the 6 minutes version the seeds germinate at reported laboratory germination is 0%
regular intervals of days (daytime), in variant 4 It should be borne in mind that ultrasound
only two of the readings are found to have stimulation with a certain duration, depending
sprouted seeds, with the difference between the on the culture, may also have a negative effect.
first and the second being 6 days.
407
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Influence of ultrasonic stimulation on the growth
Cholakov D., 2009. Zelenchukoproizvodstvo, and proliferation of Oryza sativa Nipponbare callus
Akademichno izdatelstvo na AU-Plovdiv, с. 239- cells, Colloids and Surfaces B: Biointerfaces, 27 (4),
252. 287-293.
Davidov G.K., 1940. Act of ultrasound on seeds of a Machikowa T., Kulrattanarak T., Wonprasaid S., 2013.
sugar beet. DAN USSR 29: 491-493. Effects of ultrasonic treatment on germination of
Esfandiar F., Noroozi1 H., Farbod M., Gerami1 F., 2012. synthetic sunflower seeds. Proceedings of World
Assessment of Fennel (Foeniculum vulgare) seed Academy of Science, Engineering and Technology.
germination characteristics as influenced by No. 73. World Academy of Science, Engineering
ultrasonic waves and magnetic water. European and Technology (WASET).
Journal of Experimental Biology, 2 (3): 662-666. Murtazov Т., Minkov Il., 1984. Zelenchukoproizvodstvo
Gordon A.G., 1963. The use of ultrasound in agriculture. sys selekciya I semeproizvodstvo. Izdatelstvo
Ultrasonics, 1 (2), 70-77. „Hristo G. Danov“. Plovdiv.
Istomina O., Ostrovsky E., 1936. The effect of ultra- Yaldagard M., Mortazavi S.A., Tabatabaie F., 2008.
sonic vibrations on plant development. Compt. rend. Application of ultrasonic waves as a priming
acad. sci. URSS 2: 155-160. technique for accelerating and enhancing the
Kwiatkowska B., Bennett J., Akunna J., Walker G.M., germination of barley seed: optimization of method
Bremner D.H., 2011. Stimulation of bioprocesses by by the taguchi approach. J. Inst. Brew., 114 (1), 14-
ultrasound, Biotechnology Advances, 29 (6), 768- 21.
780.
408
DETERMINATION OF CHLORINE CONCENTRATION

AND PRODUCTIVITY IN SOME BEAN GENOTYPES,
FROM NORTH-EAST OF ROMANIA, UNDER SALT STRESS
Beatrice Alexandra MODIGA, Carmenica Doina JITĂREANU, Cristina SLABU,

Alina Elena MARTA, Mihaela COVAȘĂ
„Ion Ionescu de la Brad” University of Agricultural Sciences and Veterinary Medicine of Iași,
Faculty of Agriculture, Department of Biological Sciences, Mihail Sadoveanu Alley, no. 3, 700490,
Iași, România
Abstract
From the environmental stress factors, salinity is one of the most important causes which affects the security of
mankind. Impact of soils with excessive salinity on the productivity of different cultures is sometimes disastrous, which
determines the identification and the creation of new genotypes of plants tolerant to osmotic stress conditions. From the
viewpoint of stress concept, chlorine (Cl-) is viewed as being biologically aggressive osmolite, based on its small ionic
diameter and its strong tendencies to attract water (high hydration capacity). High concentrations of these ions in
apoplast lead to imbalances in hydric and ionic relationships. For these reasons, stress caused by salinity is also a
dehydration stress and ionic stress. In present study, it was pursued the influence of saline solutions of 100 mM and 200
mM NaCl on Cl- concentration and productivity, to 7 local populations of common beans (Phaseolus vulgaris L.)
collected from areas with saline soils, in North-East of Romania and grown under greenhouse conditions, at the pots,
analyzing the largest/lowest amount of chlorine in the leaves and the correlations between the chlorine content and the
average number of pods/plants, the average number of grains/pods, the average weight of the grains/pods and the
average yield/plant, indicators that define productivity.
Key words: correlations, NaCl, Phaseolus vulgaris L., productivity.
INTRODUCTION reduces the ability of plants to utilize water,

causes a reduction in the growth and yield, and
Excess salinity in the soil affects total growth changes in the plant metabolic processes
and yield and the impact extent depends on the (Munns, 1993, 2002).
degree of salinity (Flowers and Yeo, 1995; The chloride ion is present in abundance almost
Munns, 2002). Crop plants will not grow in everywhere in the world. It is required as a
high concentrations of salt: only halophytes micronutrient for optimal plant growth, at a rate
grow in concentrations of sodium chloride of only 0.3-1 mg/g dry matter in most plants
higher than about 400 mM. The physiological (Marchner, 1986). The influence of the chloride
and molecular mechanisms of tolerance to ion on plant growth depends on the plant
osmotic and ionic components of salinity stress variety (Tottingham, 1919). The dependence of
are reviewed at the cellular, organ, and whole- modern agriculture on irrigation and chemical
plant level. Salinity is considered one of the fertilization causes more concern about the
major factors affecting the agricultural toxicity of chloride than about Cl- deficiency
productivity worldwide. In the arid and (Marschner, 1986).
semiarid regions, soil salinization may be Generally, most non woody crops tolerate
caused by poor irrigation water which contains excessive levels of Cl-, whereas many woody
considerable amounts of salts, salt plants species and beans are susceptible to Cl-
accumulation in the soil surface layer due to toxicity (Maas, 1986).
over-irrigation, proximity to the sea and/or the Chloride toxicity in plants is often hard to
capillarity rise of salts from underground water, diagnose, for two reasons: it is difficult to
into the root zone, due to the excessive separate the effects of chloride from those of
evaporation (Gama et al., 2007). Salinity any accompanying cations, commonly sodium;
409
and it is difficult to distinguish between the from the North-Eastern region of Romania,
specific toxic effects of ions and the cellular known as the region of Moldova (Iași, Vaslui
dehydration caused by their excessive external and Botoșani).
concentrations (Bar et al., 1997). The bifactorial experience was set up in 12 liter
Chlorine is involved in photosynthesis to seed pots, in randomized blocks with three
remove harmful oxidants from photochemical repetitions. They were exposed to saline stress
systems and stimulate electron transport. It has for a period of 30 days, being constantly
a role in regulating the osmotic potential by watered with concentrations of 100 mM NaCl
maintaining the hydric cell level, ensuring the and 200 mM NaCl.
opening of the stomata. On the other hand, it The concentration of chlorine was determined
stimulates enzymatic activity and is antitranspi- by potentiometric titration with silver ions,
rant (Davidescu, 1988). In the chloroplast, the using the Analytical Chloride Titrator, as
Cl- concentration remains relatively constant described by Slabu et al. (2009), and expressed
regardless of whether the plant growth medium as mg/100 g DW.
is characterized by deficienty or excess. The results were statistically interpreted using
Excess of chlorine negatively affects plant the Microsoft Excel- Data Analysis application,
growth, by degradation of carbohydrate determining the correlation coefficient and
metabolism. Species resistant to excess magnitude of the effect in the linear correlation
chlorine are: beans, potatoes, tomatoes etc. r (Pearson).
(Șumălan, 2004). Chlorine deficiency reduces
foliar growth, followed by wasting, chlorosis, RESULTS AND DISCUSSIONS
brunification, and ultimately necrosis; at the Analysis of the Cl- content of bean leaves
same time, the fruit decreases in number and subjected to saline stress, over a period of 30
size, because beans are a sensitive plant from days reflects the fact that in the case of the
this point of view (Toma et Jităreanu, 2007). control variant, values ranged from 0.15 to 0.48
Phaseolus vulgaris L. is a salt - sensitive mg/100 mg DW. For the variant treated with
species. For this reason, the purpose of the 100 mM NaCl, the values were higher
present paper was to determine the effect of compared to the control variant, so the
excess NaCl on production as an indicator of maximum was recorded at the Moșna genotype
salt stress tolerance. (6.60 mg/100 mg DW) and the minimum value
at the Blăgești 2 (5.25 mg/100 mg DW).
MATERIALS AND METHODS The values are significantly higher than the
The experience was done under greenhouse control variant and for the 200 mM NaCl stress,
conditions and the research took place in the genotypes with a maximum of 12.10 mg/100
Plant Physiology Laboratory, USAMV Iași. mg DW to the local population Blăgeşti 1,
The biological material was represented by which denotes the resistance of this genotype to
seven local bean populations (Blăgeşti 1, saline stress (Table 1).
Blăgeşti 2, Blăgeşti 3, Blăgeşti 4, Moşna,
Săveni, Trușeşti 2) collected from saline soils
Table 1. The content of Cl- (mg/100 mg DW) of bean leaves under salinity stress for a period of 30 days
Population Control 100 mM NaCl 200 mM NaCl

Blăgești 1 0.23 6.35 12.10
Blăgești 2 0.48 5.25 5.45
Blăgești 3 0.22 6.40 10.10
Blăgești 4 0.29 6.60 7.15
Moșna 0.34 6.95 9.75
Săveni 0.15 5.40 7.95
Trușești 2 0.19 5.50 5.75
410
As a result of the T-test, it is noted that there concentration of 200 mM statistical
are insignificant statistical differences differences are very significant, and
between the control variant and the variant statistically significant values were recorded
subjected to 100 mM NaCl; between the between variants treated with the two saline
control variant and the one with a NaCl solutions (Table 2).
Table 2. Statistical differences between control (I) and variants treated with saline solutions:
100 mM (II) and 200 mM (III) in terms of Cl- content, expressed as mg/100 mg DW
Comparative variants t-stat P two-tail Signification
Control -100 mM NaCl (I) -22.5725 2.474255 Ns
Control - 200 mM NaCl (II) -8.58179 0.000006 ***
100 mM NaCl - 200 mM NaCl (III) -2.87781 0.014069 *
-Test Paired Two Sample for Means: Ns-statistically insignificant differences (p≥0.05) between variants; *Significant
statistical differences (p≤0.05) between variants; **Significant distinct statistical differences (p≤0.01) between variants;
***very significant statistical differences (p≤0.001) between variants.
Assessing the influence of saline stress on plant 100 mM NaCl and 200 mM NaCl solutions the
production is a very important aspect in average number of pods/plant was low.
determining how salinity affects productivity, From table 3 it can be observed that the highest
and that is why the fructification process has genotype of the analyzed character is the
been studied on the basis of determinations of number of pods/plant is Blăgești 4, which
different indices: average number of records a number of 25.33 pods/plant in the
pods/plant, average number of grains/pods, control variant, 24 pods/plant for the variant
average weight of grains/pods and average 100 mM NaCl and 23.66 for the
production/plant. 200 mM NaCl variant. The lowest number of
Analyzing the average number of pods/plants pods/plants was recorded in the Moșna:
in saline-stressed bean plants over a period of 3.33 pods/plant for the control variant,
30 days, it is noted that, in comparison to the 1.66 pods/plant for the 100 mM NaCl
control variant, for all genotypes treated with (Table 3) .
Table 3. Effect of saline stress on the average number of pods/plant after 30 days exposure to saline stress

Blăgești 1 15.33 11.33 8.00
Blăgești 2 25.00 18.66 12.00
Blăgești 3 7.00 4.00 3.00
Blăgești 4 25.33 24.00 23.66
Moșna 3.33 1.66 0.00
Săveni 3.33 4.00 1.00
Trușești 2 15.33 9.00 6.00
The correlation between Cl- content and concentration at leaf level had a negative
average number of pods/plants after 30 days of influence on the average number of
exposure to saline stress was found to be grains/pods, in this respect the excess of
acceptable in accordance with the rules chlorine having a toxic effect (Figure 1).
established by Colton (1974). Chlorine
411
30
25
y = -0.748x + 1.418
Cl- (mg/100 mg)

20 R² = 0.107
r= 0.3271
15
10
0
0 2 4 6 8 10 12 14
Number of pods
Figure 1. Correlation between Cl- (mg/100 mg DW) and average number of pods/plant
and after 30 days exposure to saline stress
Regarding the determination of the average 200 mM NaCl variant, was particularly
number of beans/pods it is noted that compared highlighted.
to the control variant, all local populations The lowest value of this analyzed parameter
treated with 100 mM NaCl and 200 mM NaCl was recorded in the Mosna genotype: 1.72
solutions determined a decrease of the analyzed number of grains/pod for the control variant,
parameter. 1.25 number of grains/pod for the 100 mM
Thus, the Blăgeşti 4 genotype, which had a NaCl variant and number of pod/plant for the
number of 6.03 number of grains/pod in the 200 mM NaCl treated variant (Table 4).
control variant, 5.61 number of grains/pod for
the 100 mM variant and 4.06 grains/pod for the
Table 4.Effect of saline stress on the average number of grains/pod after 30 days of exposure to saline stress

Blăgești 1 3.56 3.16 2.72
Blăgești 2 4.42 3.19 2.72
Blăgești 3 2.72 2.26 2.16
Blăgești 4 6.03 5.61 4.06
Moșna 1.72 1.25 0.00
Săveni 3.31 3.00 1.33
Trușești 2 4.20 3.54 2.40
The correlation between Cl- content and the level negatively influenced the average number
average number of grains/pod after 30 days of of grains/pod, from this point of view
exposure to saline stress has shown an the excess of chlorine having a toxic effect
acceptable degree of association, indicating that (Figure 2).
the chlorine concentration recorded at the leaf
412
30 y = -0.749x + 1.418
R² = 0.107
25
r= 0. 3271
Cl- (mg/100 mg DW)

20
15
10
0
0 2 4 6 8 10 12 14
Grains/pod
Figure 2. Correlation between Cl- (mg/100 mg DW) content and average number of grains/pod
after 30 days exposure to saline stress
Determinations by the influence of saline stress weight of the grains/pod was the same, as in the
on weight grains/pod after 30 days of saline 100 mM NaCl treated variant. And this time the
stress, in the case of the 200 mM NaCl variant, Moșna genotype was noted for lack of
four genotypes recorded lower values, except productivity (Table 5).
the Blăgești 3 genotype at which the average
Table 5. Effect of saline stress on average weight of grains/pod after 30 days exposure to saline stress
Blăgești 1 0.82 0.56 0.37
Blăgești 2 1.10 0.96 0.73
Blăgești 3 0.56 0.32 0.32
Blăgești 4 1.85 1.70 1.30
Moșna 0.39 0.31 0.00
Săveni 0.63 0.54 0.33
Trușești 2 1.36 1.22 0.74
The correlation between Cl- content and exposure to saline stress also indicates an
average weight of grains/pod after 30 days of acceptable degree of association (Figure 3).
30
25
Cl- (mg/100 mg DW)
20
y = -0.167x + 3.833
R² = 0.197
15
r=0.443
10
0
0 2 4 6 8 10 12 14
Grain weight
Figure 3. Correlation between Cl- content and average weight of grains/pod after 30 days exposure to saline stress
413
Concerning in the mean production per plant variant (46.86 g) and for the saline-treated
compared to the control variant, it is observed a variants (40.80 g and 30.75 g).
decrease with the application of saline The smallest production was recorded in the
treatments to all seven studied.genotypes. Moșna genotype, from the 100 and 200 mM
It was found that the highest production of the NaCl variants, namely 0.51 and 0 grains/plant
Blăgeşti 4 genotype, both for the control (Table 6).
Table 6. Effect of saline stress on average production/plant after 30 days exposure to saline stress

Blăgești 1 12.57 6.34 2.92
Blăgești 2 27.50 17.91 8.76
Blăgești 3 3.92 1.28 1.28
Blăgești 4 46.86 40.80 30.75
Moșna 1.29 0.51 0.00
Săveni 2.09 2.16 0.33
Trușești 2 20.84 10.98 4.94
Regarding the correlation between Cl- content exposure to saline stress also indicates an
and average production/plant after 30 days of acceptable degree of association (Figure 4).
30
25
y = -1.154x + 1.723
Cl- (mg/100 mg DW)
R² = 0.094
20
r= 0.306
15
10
0
0 2 4 6 8 10 12 14
Production/plant
Figure 4. Correlation between Cl- (mg/100 mg DW) content and average production/plant
after 30 days exposure to saline stress
Excess chlorine accumulated in plants the ability to reduce chlorine transport in stems
following NaCl treatments at 100 mM and 200 and leaves.
mM, respectively, has a negative influence on This character is genetically determined and
the average number of pods/plants, the average can be used to improve resistance to toxicity.
number of grains/pods, the average weight of The results obtained are in full agreement with
the grains/pods and the average yield/plant, in those presented in the literature, according to
which case chlorine becomes toxic to plants, which the high concentrations of Cl- negatively
inhibiting the fructification process. The influence the production (Singh et al., 2012).
differences in susceptibility to toxicity between
the bean genotypes studied are correlated with
414
CONCLUSIONS Davidescu D., 1988. Microelemente în agricultură.
Editura Academiei R.S. România.
Flowers T.J., Yeo A.R., 1995. Breeding for salinity
Cl- content values were higher than control resistance in crop plants - where next? Australian
variant for genotypes subjected to a 200 mM Journal of Plant Physiology, 22: 875-884.
NaCl. The maximum value reached was 12.1 Flowers T.J., 1988. Chloride as a nutrient and as an
mg/100 mg DW in the Blăgești 1, and the osmoticum. Adv. Plant Nutr. 3: 55-78.
minimum value was 5.75 mg/100 mg DW at Gama P.B.S., Inanaga S., Tanaka K., Nakazawa R.,
2007. Physiological response of common bean
Trușești 2. The Blăgeşti 4 genotype recorded (Phaseolus vulgaris L.) seedlings to salinity stress.
the highest production in all four indices Afr. J. Biotechnol., 6 (2): 79-88.
analyzed, and the lowest values were registered Maas E.V., 1986. Physiological responses to chloride. In
on Moşna local population, with the Special Bulletin on Chloride and Crop Production
specification that the variant treated with 200 (T.L. Jackson, ed.). Potash & Phosphate Institute.
Atlanta, GA. No. 2: 4-20.
mM NaCl did not have any production. Marschner H., 1986. Mineral Nutrition of Higher Plants.
The fructification process decreased compared Send ed., pp. 299-312, 296-404. Academic Press, San
to the control variant of all treated genotypes Diego, C.A.
with 100 mM NaCl and 200 mM NaCl. The Munns R., 1993. Physiological processes limiting plant
correlation between the Cl- content and the growth in saline soils: some dogmas and hypotheses.
Plant, Cell and Environment, 16:15-24.
average number of pods/plant, the average Munns R., 2002. Comparative physiology of salt and
number of grains/pods, the average weight of water stress. Plant, Cell and Environ., 25: 239-250.
grains/pods and the average yield/plant in bean Singh J., Divakar Singh J., Divakar Sastry E.V., Singh
plants exposed to saline stress for a period of V., 2012. Effect of salinity on tomato (Lycopersicon
30 days from exposure to saline stress was esculentum Mill.) during seed germination stage.
Physiol Mol Biol Plants, 18 (1): 45–50.
found to be acceptable in accordance with the Slabu C., Zorb Ch., Steffens D., Schubert S., 2009. Is salt
rules established by Colton (1974). stress of faba bean (Vicia faba) caused by Na+or Cl–
toxicity? Journal of Plant Nutrition and Soil Science
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Şumălan R., 2004. Fiziologia plantelor. Elemente de
Bar Y., Apelbaum A., Kafkafi U., Goren R., 1997. fiziologia plantelor regenerate şi reacţie la stres.
Relationship between chloride and nitrate and its Editura Eurobit, Timişoara.
effect on growth and mineral composition of avocado Toma L.D., Jităreanu C.D., 2007. Fiziologia plantelor.
and citrus plants. J. Plant Nutr. 20: 715-731. Editura ,,Ion Ionescu de la Brad”, Iași.
Colton T., 1974. Statistics in Medicine. Boston: Little, Totingham W.E., 1919. A preliminary study of the
Brown and Company; 211. influnce of chlorides on the growth of certain
Cuartero J., Munoz R.F., 1998. Tomato and salinity. agricultural plants. J. Am. Soc. Agron. 11: 1-32.
Scientia Horticulturare, 78 (1-4): 83-125.
415
416
STUDY OF THE INFLUENCE OF A BIOSTIMULATOR USED

IN THE TREATMENT SEED OF FAMILY APIACEAE BY ULTRASOUND
Irena DAKOVA, Nikolina SHOPOVA, Manol DALLEV, Vangel CHILINGIROV,

Galya HRISTOVA, Ivan MITKOV
Abstract
Root vegetables - carrot, parsley and celery are crops of great economic importance due to their high nutritional and
biological value. The article explores the effect of biostimulator and the different duration of ultrasound treatment on
the sowing quality of carrot, parsley and celery seeds. From the studies done, it was found that the biostimulator effect
differs for the three cultures and depends on the duration of the ultrasound treatment.
Key words: biostimulator, rood vegetables, seeds, ultrasound.
INTRODUCTION
Root vegetables - carrot, parsley and celery are
very important crops for economic and The survey was conducted in the period 2016-
nutritional reason. 2017 in the Department of Mechanization and
Carrots (Daucus carota), which are the main in the Department of Gardening at the
root crop, are used both for fresh and canned Agricultural University of Plovdiv. The subject
consumption (Cholakov, 2009). They refer to of the study was the carrot, parsley and celery
polyvitamins vegetables containing vitamins seeds. An ultrasonic bath is used to perform the
B1, B2 and C. The content of carotene is study (Figure 1)..
between 5-15 mg per cent. Besides food,
carrots are used in pharmacy and perfumery
industry.
The harvest of early production occurs during
the spring and summer months and is
consumed exclusively in a fresh state. The late
Polish production, which is essential for our
country, has the task of satisfying the needs of
the canning industry and the population during
the winter months.
With the content of nutrients and especially of Figure 1. BRAND CT-405
essential oils, celery takes one of the first
places among vegetable cultures used as The bathtub works on the principle of intense
seasonings. The importance of celery stands out sound pressure, created by an ultrasound
particularly well given the fact that root crops source, filled in a limited volume of liquid - a
are excellent and can be used fresh during the number of microscopic bubbles are formed that
winter and spring months. (Liu et al., 2003) shrink and expand in sync with the frequency at
The aim is investigating the impact of the which they hit the surface of the objects placed
biostimulator used in seed treatment by in the liquid.
ultrasound (Istomina et al., 1936; Esfandiar et Two exposures of ultrasound treatment were
al., 2012; Gordon et al., 1963) on the seed tested at 6 and 12 minutes at a frequency of 43-
qualities of carrot, parsley and celery. 45 kHz. Two different liquids were used to
417
perform the ultrasound treatment. The first one 3) On the 21st day for celery.
was distilled water, and the second in the The germinating energy, expressing the
distilled water was added a Winner biosti- percentage of normally germinated seeds under
mulator at a concentration of 1: 1000. optimal conditions, but for a shorter period of
The biostimulator Winner has the following time, was reported to:
composition: nitrogen 125 g/l, phosphorus 1) 7th day for carrots (under BDS);
75.2g / l, potassium 54.5 g/l, calcium 0.5 g/l, 2) 10th for parsley (under BDS);
magnesium 7.5 g/l, iron 0, 5 g/l. It also contains 3) 10th for celery (under BDS).
manganese, copper, zinc, sulphur, pine and
molybdenum. It is rich in organic substances - RESULTS AND DISCUSSIONS
amino acids, hormones, vitamins - 57.2%. It is
applied to foliage and soil, but it also serves to 1. Germination energy (%)
treat seeds in different concentrations The results of the performed studies show an
depending on the species. increase in exposure and with the addition of a
Non-treated seeds of the indicated crops plan- biostimulator, the percentage of germinating
ted for germination in petri dishes were used energy increases.
for the control. Depending on the duration of Highest test values were recorded in the 12
the seed treatment and the used liquid, the minutes seed treatment scenarios where the
following variants were tested when loading germinating energy reaches 100% and the
the ultrasonic bath: excess over the control was 233.3%. At 6
Carrot: minutes exposure, the germinating energy
1. Control; increased to 85%.
2. Exposure 6 min. + Winner; The lowest values were recorded in the control
3. Exposure 12 min. + Winner; - 30% sparying energy (Figure 2).
Parsley:
1. Control;
2. Exposure 6 min. + Winner;
Celery:
1. Control;
3. Exposure 12 min. + Winner.
The experiment was set in 3 replicates and the
number of seeded counts in each iteraction was
100. After the treatment, the seeds were placed
in a thermostat at 24°C, with the number of
seed germinated daily. The counting continued Figure 2. Germinating energy (%) of carrot seed, average
for the period 2016-2017
until the 14th day for carrot, 21 for celery and
28 for parsley.
While the germinating energy increases in
Two parameters were tested - laboratory
carrots with the addition of biostimulants, the
germination (%) and germinating energy (%).
effect of the studied effects is less pronounced
As a result of day-to-day readings, the speed
in parsley and celery.
and cooperativity of seed germination were
The germinating energy recorded in parsley
determined.
(Figure 3) increases, although slightly less than
Laboratory germination, showing % normal
20% (seen in the control) and reaches 30%
germinated pure seeds under optimum
when the biostimulator is added.
conditions for a specified period of time, was
Exceedance of the control for this option is
determined by counting the number of
50%.
germinated seeds according to the accepted
BDS standards (601 84) as follows:
1) On the 14th day for carrot;
2) On the 28th day for parsley;
418
And with this main indicator, the trend found in
carrots to enhance the effect of adding a
biostimulator is retained (Figure 5).
The reported germination exceeds that of the
control by 58.7%. Within 6 minutes exposure,
germination reached 95%, and the over-control
was 50.8%. Significantly lower than the other
variants is germination, reported in the control -
63%.
Figure 3. Germination energy (%) of parsley seeds

averaged over the period 2016-2017
The treatment of celery seeds with ultrasound

(Figure 4) affects germination energy only
when its duration is 12 minutes.
Within this exposure, the measured values of
the test item do not differ significantly between
each other and the difference between the two
tested media is only 5%, with the biostimulator
variant being 19%. Figure 5. Laboratory germination (%) of carrot seed,
In the other variants, including the control, 0% averaged over the period 2016-2017
of germinating energy was reported.
Attention is paid to the results presented in
Figure 6, providing information on the speed
and suitability of seed germination in carrots. It
can be seen from the figure that, with or
without biostimulator, in the first 3 days after
this phase occurs, the number of germinating
seeds depending on the variant is between 55
and 100. For comparison, in the control for the
same period of time the number of sprouts is
30.
In the biostimulator variant on Day 4, 60
Figure 4. Germination energy (%) of celery seeds, germinations were reported and the remaining
averaged over the period 2016-2017 40 on the 5th day. The biostimulator effect in
the 6 minutes exposition was also well-known,
2. Laboratory germination (%) whereas on day 6 (3 days after start of
Germination is a very important indicator of a germination) the number of sprouted seeds
biological aspect that provides information on recorded is 27.3% higher than the other variant
the viability of the seeds, their suitability and with the same treatment duration.
the determination of the seed norm (Murtazov,
1984).
419
Figure 6. Germination of carrot seeds by day
The most cumbersome and fastest germinating

seeds are at 12 minutes exposures, with 60
germination on day 4 and the remaining 40 on
day 5.
The biostimulator effect is also well-known in
6 minutes exposure, on the 6th day (3 days after
germination) the number of germinated seeds
recorded was 27.3% higher than the other
variant with the same treatment.
In Figure 7 shows the data on laboratory
germination of parsley seeds. Figure 7. Laboratory germination (%) of parsley seeds
The highest percentage of laboratory averaged over the period 2016-2017
germination was reported in the 6 minutes
exposition test, with the biostimulator reading The results of Figure 8 it is noted that seed ger-
being 90%. mination begins from day 6 until day 11, at the
The difference between the two 6 minutes earliest 6 minutes biostimulator exposition and
exposure variants is 10%. In the control, the at 12 minutes exposure at the latest. The control
germination rate was 60%. sprout starts on the 9th and ends on the 14th.
Figure 8. Sprouting seeds of parsley by day for the whole period
420
In Figure 9 the results for laboratory As far as the biostimulator is concerned, it has
germination of celery are presented. Under the a negative effect, because in both embodiments
influence of ultrasound stimulation germination involving this additive there are reported
of seeds strongly increases. significantly lower results compared to the
At 0% of the control, laboratory germination distilled water variants.
increases with increasing treatment duration It is noteworthy that even here the tendency to
and reaches 70% in the 12 minutes exposure increase germination with an increase in ultra-
scenario. 6% lower is the germination reported sonic processing is maintained. Although 9% of
in option 2 (6 minutes). the benefit is for the 12 minutes option.
Figure 9. Laboratory germination (%) of celery seeds, average for the period 2016-2017
The germination process (Figure 10) starts at day 13), the number of sprouts at each
the earliest with 12 minutes exposures - 8th day subsequent counting increases and reaches it’s
for variant 3 (12 min.) and 9th day for variant 5 maximum on the 17th day - 35 pieces.
(12 min. + Winner). In the same two variants This option is the shortest and the start-end
seed germination ends on day 17 on day one period of germination - 5 days, for which 64
and on day 18 in the second variation. sprouted seeds have been reported. When
The first germinated seeds in the 6-minute compared to option 3 (12 minutes), where the
exposure variants were counted on the 13th day highest germination rate was recorded, it is
after treatment, with no new germinated seeds seen that the period is 10 days. Apparently
being reported for the biostimulator addition 6 minutes seed treatment with ultrasound helps
until 19th day. Interestingly, the results are to seed the seeds more comfortably, and this is
reported for option 2. It is clear from the graph essential for obtaining quality and even plants.
that, despite the later onset of germination (on
421
Figure 10. Germination of celery seeds by day for the whole period
CONCLUSIONS Esfandiar F., Noroozi1 H., Farbod M., Gerami1 F., 2012.
Assessment of Fennel (Foeniculum vulgare) seed
germination characteristics as influenced by ultrasonic
The biostimulator effect is different for the waves and magnetic water. European Journal of
three cultures and depends on the duration of Experimental Biology, 2 (3): 662-666.
the ultrasound treatment. Gordon A.G., 1963. The use of ultrasound in agriculture.
In carrots and parsley, the addition of Ultrasonics, 1 (2), 70-77.
biostimulator increases germination by up to Istomina O., Ostrovskiy E., 1936. Effect of ultrasound on
evolution of plants. DAN USSR, New series 2, 155
13% in carrots and by up to 20% in parsley. (in Russian).
It should be borne in mind that adding a Liu Y., Yoshikoshi A., Wang B., Sakanishi A., 2003.
biostimulator can also have a negative effect. Influence of ultrasonic stimulation on the growth and
proliferation of Oryza sativa Nipponbare callus cells,
REFERENCES Colloids and Surfaces B: Biointerfaces, 27 (4), 287-
293.
Murtazov Т., Minkov Il., 1984. Zelenchukoproizvodstvo
Cholakov D., 2009. Zelenchukoproizvodstvo, sys selekciya I semeproizvodstvo. Izdatelstvo „Hristo
Akademichno izdatelstvo na AU-Plovdiv, 239-252. G. Danov“. Plovdiv.
422
EFFECT OF FOLIAR BIOACTIVE TREATMENTS ON THE

OXIDATIVE STRESS TOLERANCE IN TOMATO SEEDLINGS
Gabriela LUŢĂ, Daniela BĂLAN, Evelina GHERGHINA, Elena DOBRIN

Abstract
Under the current climate changes, enhancing temperature is now considered to be one of the major abiotic stresses
inducing negative effects on plant growth as results of increased production of toxic reactive oxygen species. The
tolerance of stress is correlated with higher activities of antioxidant defense enzymes which are activated to prevent
oxidative damage. On the other hand, the use of biofertilizers appears to be involved in increase of tolerance to various
abiotic stresses, besides their important role in regulating plant growth, development and fruiting. Influence of
environmental conditions and of some growth regulators treatments (Spraygard 1%, Razormin 0.1%, BAC Foliar spray
0.3% and BIO Roots 0.2%) on the chlorophylls and carotenoids content and on the activity of peroxidase and catalase
have been investigated in leaves of tomato seedlings. Analysis of the obtained data emphasize the potential of growth
regulators in enhancing the resistance to abiotic stresses by protecting the photosynthetic apparatus and improving the
activity of antioxidant enzymes system.
Key words: carotenoids, catalase, chlorophylls, heat stress, peroxidase.
INTRODUCTION Recently some countries practice tomato

growing even at high temperature through
Tomato (Solanum lycopersicum) is considered application of plant growth regulators. Several
one of the most important vegetable whose authors reported that application of substances
production is constantly growing since its fruits like auxin, gibberellic acid, synthetic auxin 4-
are widely consumed either fresh or processed. CPA (4-chloro phenoxy acetic acid) have
Beside the high nutritional value, the ripe resulted in good tomato production under
tomato fruits are a valuable source of vitamin adverse environmental conditions (Sasaki et al.,
C, lycopene, carotenoids and minerals such as 2005; Gemici et al., 2006; Khan et al., 2006;
iron and phosphorous that are daily required for Poliquit et al., 2007; Gelmesa et al., 2012).
a healthy diet (Nour et al., 2013). At present, Also good results in improving freezing
the tomatoes acquired great popularity among resistance by growth regulators as Ruter AA,
consumers, being considered a protective food Terra Sorb and Razormin were noticed in
since the discovery that lycopene has anti- experiment with winter wheat cultures
oxidative and anti-cancer properties (Gajowik, (Gaveliene et al., 2016).
2014; Raiola et al., 2014). Besides their important role in improvement of
Although tomato plants can grow under a wide nutritional quality of food crops and their
range of climatic conditions, they are extremely efficiency to regulate plant growth,
sensitive to growing conditions: high development, fruiting and senescence (El-
temperature (both day and night), humidity, Rokiek et al., 2012), it seems to be involved in
rainfall and light intensity are the limiting induction of tolerance to various abiotic
factors of tomato production (Ahmad, 2002). stresses (Salehi et al., 2011; Gaveliene et al.,
Temperature stress induces negative effects on 2016).
plant growth and metabolism, so high Plants exposed to extreme temperatures
temperature is now considered to be one of the activated the self-defense mechanisms
major abiotic stresses causing yield reduction including several non-enzymatic and enzymatic
in crops (Hasanuzzaman et al., 2012). antioxidants (-tocopherol, carotenoids,
chlorophylls, glutathione, ascorbic acid,
423
oxidases such as catalase, peroxidase, Experimental variants were noted:
polyphenol peroxidase, superoxide dismutase) T0 - unstressed control;
to prevent oxidative damage (Vranova et al., T1 - tomato seedlings exposed at heat stress
2002; Torres-Barceló et al., 2013). Some in absence of growth regulators treatment;
scientific works reported that higher activities T2 - tomato seedlings exposed at heat stress
of antioxidant defense enzymes are correlated and treated with Spraygard 1%;
with higher stress tolerance (Almeselmani et T3 - tomato seedlings exposed at heat stress
al., 2006; Babu et al., 2008; Almeselmani et al., and treated with Razormin 0.1%;
2009). T4 - tomato seedlings exposed at heat stress
This study was aimed to investigate the effect and treated with BAC Foliar spray 0.3%;
of growth regulators containing free amino T5 - tomato seedlings exposed at heat stress
acids, macro- and micro-elements (Razormin, and treated with BIO Roots 0.2%.
Spraygard, BAC Foliar spray, Bio Roots) used Specific agrotechnics for transplant nursery
as foliar treatments on tomato seedlings and the was applied during the growth period: daily
comparative results were studied. The effect of ventilation, watering, weeding. The unstressed
the bioregulators on antioxidant enzymes and control variant was maintained at the
photosynthetic pigments in tomato plant is less temperature of 22-26oC at day and 18-20oC at
known, so that the influence of environmental night. For the heat stressed variants, the
conditions and of some foliar treatments on temperature was not controlled and registered
biochemical parameters (chlorophylls and the following variations:
carotenoids content, activity of peroxidase and the maximum effective temperature
catalase) has been investigated in leaves of average for April in the greenhouse was
tomato seedlings. 31.8oC and the minimum effective
temperature average at 24.6oC;
MATERIALS AND METHODS the maximum effective temperature
average for May in the greenhouse was
The experiment was installed into an 37.4oC and the minimum effective
experimental greenhouse of the Hortinvest temperature average at 25.2oC.
Research Centre - USAMV Bucharest. The The biochemical determinations in the active
main objective was testing the effects of leaves were performed at the end of the
applications of simple growth regulators experiment (after 27 days), when most of
Razormin 0.1%, Spraygard 1%, BAC Foliar seedlings have reached the optimum for a
spray 0.3% and BIO Roots 0.2% on heat succesfull planting.
stressed and unstressed tomato seedlings in two In order to estimate the oxidative stress
distinct stages: one week, respectively two occurred on cell level, characteristic parameters
weeks after transplant operation. were analyzed, such as proteins content,
Razormin is a mixture of growth factors (amino specific activities of catalase as well as
acids, polysaccharides, macro and micro- peroxidase, assimilatory pigments content
nutrients), which induces development of the using appropriate methods of analysis.
root system, stimulate the nutrients absorption, The proteins content was determined
increases vegetative mass and quality of by Lowry method, which is based on the
production. reactivity of the peptide nitrogen with the
Spraygard is a complex product that acts as copper [II] ions under alkaline conditions and
safener, penetrant, dispersant, creates adhesion the subsequent reduction of the Folin-Ciocalteu
of the treatment solutions on the leaves. phosphomolybdic-phosphotungstic acid to
BAC Foliar is a foliar organic nutrient which heteropoly-molybdenum blue by the copper-
stimulates chlorophyll production in the leaves. catalyzed oxidation of aromatic aminoacids
Bio Roots is a natural root growth supplement (Lowry et al., 1951). The results were
which contains vitamins, enzymes, organic and expressed in g/100 g fresh weight.
humic acids that helps plants establish healthy The activity of peroxidase was
and vibrant roots. determined by spectrophotometric measuring
of the speed of colour achievement at 436 nm
424
and 25oC in the decomposition reaction of accessory components in the photosynthetic
hydrogen peroxide with guaiacol as hydrogen complex by providing photoprotection and
donor, which is catalyzed by peroxidase stability of proteins present in the photosystem
(Bergmayer, 1974). (Torres-Netto et al., 2005; Simkin et al. 2008).
Photosynthesis, one of the most heat sensitive
processes, can be completely inhibited by high
The enzymatic unit: the amount of enzyme
temperature possibly as a result of structural
which catalyzed the transformation of one
and functional disruptions of chloroplasts and
micromole of hydrogen peroxide/minute at
reduction of chlorophyll accumulation under
25oC.
high temperature stress (Camejo et al., 2001;
The activity of catalase was determined
Dekov et al., 2000).
with method essentially described by Beers and
The determinations performed on the tomato
Sizer (1952), in which the disappearance of
seedlings showed that high temperature
peroxide is followed spectrophotometrically at
affected both chlorophylls a and b, therefore
240 nm.
the total chlorophyll content. A decrease of
The enzymatic unit: one unit decomposes one
about 40% was registered in the untreated
micromole of H2O2 per minute at 25°C and pH
plants exposed to heat stress (T1) compared to
7.0 under the specified conditions.
the unexposed control plants (T0) (Figure 1).
Determinations of the assimilatory
pigments content in the active leaves:
chlorophyll and carotenoid pigments were
extracted in 80% acetone and the absorbance of
the extract was measured at three wavelengths
(663 nm, 647 nm and 480 nm) with an
UV/Visible ThermoSpectronic Helios
spectrophotometer. The results were calculated
using the extinction coefficients and equations
described by Schopfer (1989) and were
expressed in mg/100 g fresh weight (FW).
Figure 1. Assimilatory pigments content in the
experimental variants
Temperature is a major factor affecting the rate

The effect of high temperature exposure on
of plant development. Warmer temperatures
chlorophyll content has been extensively
expected with climate change will affect the
studied and similar results were reported at
physiological processes in plants, therefore the
some tomato cultivars (Berova et al., 2009) and
plant productivity. Some adaptation strategies
also in other plants as Triticum aestivum
are available to manage with temperature
(Tewari et al., 1998; Almeselmani et al., 2012),
extremes depending on the plant species, being
Festuca arudinacea (Cui et al., 2006), Solanum
genetically determinate. Beside this,
ssp. (Aien et al., 2011). Other authors have
application of some bioregulators is expected to
found an increase in chlorophyll a content in
increase tolerance to high temperature stress.
lemon stressed plants with high temperatures
So analyze of assimilatory pigments and
(Martin et al., 1995).
peroxidase and catalase activity as oxidative
A higher reducing in chlorophyll b content
stress markers was performed.
compared to chlorophyll a content (Table 1)
Determination of assimilatory pigments
was registered, so the chlorophyll a/b ratio
content
registered an increase in the stressed plants
Chlorophylls a and b represent the major
compared with control plants, in according to
photosynthetic pigments in plants, playing an
the data reported by other authors (Cui et al.,
important role in the photochemical reactions
2006; Zhu et al., 2011).
involved in photosynthesis (Taiz and Zeiger,
2009), while carotenoids are considered as
425
Table 1. Values and ratio of assimilatory pigments in the experimental variants
Variants Chlorophyll (a+b) Carotenoids Chlorophyll (a+b)/ Chlorophyll
(mg/100 g FW) (mg/100 g FW) carotenoids a/b
T0 139.66 2.52 55.42 2.34

T1 81.25 2.66 30.55 3.70
T2 122.67 3.45 35.56 2.96
T3 112.68 3.57 31.56 3.08
T4 124.67 3.77 33.07 2.92
T5 98.06 3.71 26.43 3.12
Scientific studies reported that a 10-15oC reported lower chlorophyll (a+b)/carotenoid

increase over normal growth temperature ratio in two heat stressed cultivars of Festuca
results in degradation of chlorophyll and thus arudinacea in relation to the control plants (Cui
affecting photosynthesis process. The reasons et al., 2006) and carotenoids amounts increased
for decreasing in photosynthetic pigments in Populus cathayana cuttings exposed under
under high temperature may be attributed to the moderate stress conditions (Xiao et al., 2008).
inhibition of biosynthesis, changes in However, the increase of carotenoids amounts
ultrastructure of chloroplast and photo- was more pronounced in the stressed tomato
deterioration (Tewari et al., 1998; Reda et al., plants under growth regulators treatment
2011). (between 3.45 mg/100 g FW in Spraygard
Observations and measurements performed on treated variant and 3.77 mg/100 g FW in BAC
the experimental variants under growth Foliar treated variant) compared with untreated
regulators treatment showed a smaller control (2.66 mg/100 g FW at T1). Similar
diminution of total chlorophyll content in observations of bioregulators treatment
stressed plants: about 11% in the plants treated increasing carotenoids content were reported in
with BAC Foliar (T4) and Spraygard (T2) some species of pepper plants (Capsicum
compared to the unexposed control plants (T0). annuum var. grossum, Capsicum annuum var.
Generally, it seems that the growth regulators accuminatum), in eggplant seedlings (Balan et
treatment induced a better accumulation of al., 2018), while higher bioregulator
chlorophylls in the tomato leaves. concentrations impacted negatively on
Carotenoids are an important class of carotenoid content in Capsicum chinense plants
antioxidants which play a major role in the (Olaiya et al., 2013), also in Triticum aestivum
protection of plants against photo-oxidative plants (Sahu et al., 2011). Total carotenoid
processes (Stahl et al., 2003; Gramza- content did not change with stress conditions in
Michalowska et al., 2010). Therefore, tomato cv. Amalia, while increased in
maintaining a higher or invariable level of total Nagcarlang cv. (Camejo, 2001). This results
carotenoids during stressful conditions may suggests that the response to treatments with
induces some stress tolerance of the plants the bioregulators is probably genetically
(Loggini, 1999; Logan et al., 1996; Ruban, determined.
1999). Carotenoids destruction through It is well documented that carotenoids act as
oxidation may reduce efficiency of the antioxidant compounds involved in protection
antioxidant defense system (Chedea et al., of photosynthetic systems, therefore a higher
2013). level of total carotenoids support the plant to
A slight increase of the carotenoids content was tolerate the stressful condition. These results
registered as result of heat exposure of the are in agreement with previous studies related
tomato seedlings (Table 1): 2.66 mg/100 g FW to plant acclimation to stress (Loggini et al.,
carotenoids determined in stressed control in 1999; Ruban et al., 1999).
absence of treatment (T1) compared to 2.52 Determination of content in proteins
mg/100 g FW carotenoids in unstressed control. Proteins content analyzed in the leaves of
Also a decrease of chlorophyll tomato seedlings showed that heat exposure
(a+b)/carotenoids ratio was registered in the induced an increase, so that higher values of
stressed plants (Table 1). Previous studies this parameter were registered in the stressed
426
plants (1.37 g %) in absence of bioregulators under treatment with roots stimulator BIO
treatment compared with the unstressed control Roots (0.33 U/mg protein) and with BAC
(1.25 g %) (Figure 2). However, the increase of Foliar (0.43 U/mg protein).
protein content was more pronounced in the Other authors also reported that some
leaves of the tomato treated with growth treatments of plants with plant growth
regulators (between 1.57-2.15 g %) in regulators showed positive effects on oxidases
comparison to untreated stressed control (1.37 activities. For example, methyl jasmonate-
g %), BAC Foliar treated variant (T4) reaching treated raspberries, strawberries and blueberries
the highest value of proteins amount (2.15 g showed higher activities of peroxidase and
%). Previous studies noticed that an increase in superoxide dismutase (Chanjirakul et al.,
protein content might suggest a change in the 2006).
gene expression that would be associated with a Previous studies documented that the activation
possible thermotolerance and acclimatization to of protein synthesis in plants in combination
stress condition (Camejo et al., 2001). with increase of oxidases activity under stress
conditions may be the result of metabolism
conversion in order to obtaining a good
tolerance of plant to stressful conditions. (Tucic
et al., 2007; Chkhubianishvili et al., 2011; Wu
et al., 2014).
CONCLUSIONS
The researches performed on the tomato

seedlings showed that high temperature
affected both chlorophylls a and b, therefore
Figure 2. Enzymatic activity and proteins content in the the total chlorophyll content. At the same time
experimental variants it seems that the growth regulators induced a
better accumulation of chlorophylls in the
Determination of enzymatic activity in the tomato leaves since a smaller diminution of
seedlings leaves total chlorophyll content in stressed tomato
There are numerous previous studies which plant was noticed under growth regulators
indicate that the tolerance to temperature stress treatment.
in plants may be positively correlated with an A slight increase of the carotenoids content in
increase in antioxidants content (Babu et al., the control plant occured as result of heat
2008; Almeselmani et al., 2009; Hasanuzzaman exposure but higher values of this parameter
et al., 2013). were determined in the stressed tomato plants
A slight increased activity of peroxidase was under influence of growth regulators treatment.
registered in the untreated tomato seedlings Also the protein content and the oxidases
under heat stress (0.13 U/mg protein) compared activity were enhanced under high temperature
with control (0.11 U/mg protein), but response conditions, mostly in the tomato seedlings
to heat stress was amplified in the tomato treated with growth regulators.
seedlings under growth regulators treatment by An overview of the researches performed on
enhancing the peroxidase activity (between the tomato seedlings allows us to conclude that
0.17 U/mg protein in Razormin variant and growth regulators treatments determined an
0.31 U/mg in BAC Foliar variant). increased plant capacity to face the effects of
Catalase activity follows the same dynamics as heat stress by protecting the photosynthetic
peroxidase: 0.09 U/mg protein in control apparatus and enhancing antioxidant enzyme
tomato seedlings, which increased at 0.12 systems. Good results in this sense were
U/mg protein in absence of bioregulators. Also obtained mainly with BAC Foliar spray and
the catalasic activity increased in the stressed Razormin, which proved the best ameliorative
plants treated with growth regulators. Higher effect under heat stress conditions.
catalase activities were registered in the tomato
427
However, further studies should be considered Chedea V.S., Jisaka M., 2013. Lipoxygenase and
in order to asses possible combined treatments carotenoids: A co-oxidation story. African J.
Biotechnol, 12 (20): 2786-2791.
with several growth regulators to achieve Chkhubianishvili E., Kacharava N., Badridze G.,
optimum effects in improving the plants growth Chanishvili S., Kurdadze T., 2011. Activity of
and productivity under environmental stress in Peroxidase, Catalase and Content of Total Proteins in
the conditions of global climate change. Leaves of some Herbaceous Plants of High
Mountains of the Caucasus, Bull. of the Georgian
National Academy of Sciences, 5 (2): 96-100.
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430
DETERMINATION OF THE RESPONSES OF DIFFERENT

TOMATO SPECIES TO TUTA ABSOLUTA
Ali KAYAHAN1, Betül SIMSEK1, Ismail KARACA1, Hakan AKTAS2

1
Suleyman Demirel University, Faculty of Agriculture, Plant Protection Department, Isparta/Turkey
2
Suleyman Demirel University, Faculty of Agriculture, Horticulture Department, Isparta/Turkey

Abstract
In this study, the harmful effect of Tuta absoluta on different tomato species was investigated. Six tomato genotypes
which are Solanum chmielewskii (LA1028), Solanum neorickii (LA0247), Solanum hirsutum (LA777), Solanum
pimpinellifolium (LA0722), and Solanum lycopersicum cv. Cuatomate and Solanum lycopersicum cv. Ailsa Craig
(LA2838A) species were used as plant material in the study. To determine the incidence of Tuta absoluta on the plants,
the egg and larvae were counted weekly by taking 5 leaves from the species. According to eggs and larvae counts, the
plants were exposed to 5 types of severe damage while Solanum lycopersicum cv. Cuatomate was found to be later and
less effective than the others. These results indicate that this species exhibits antixenosis properties against Tuta
absoluta pests and that the other species that have been exposed to earlier and more severe damage lack such property.
Key words: Tuta absoluta, Solanum species, antixenosis.
INTRODUCTION tomato leaf miner is one of the major

devastating pests of processing and fresh
Tomato (Solanum lycopersicum L.) is one of tomatoes, both in greenhouse and open field
the most important food crops in the world in condition. Tuta absoluta larvae can absolutely
terms of human consumption, while total global destroy the tomato canopy by excavating the
production exceeds 5 million hectares leaves, stems and buds; and burrows into fruits
worldwide with an average annual production causing the quality decline of fresh tomato and
of more than 170 million metric tons (FAO, yield loss (Cocco et al., 2012). It is an
2017). However, declining agricultural areas oligophagous insect that feeds on at least 39
and increasing pests and diseases are affecting species belonging to ten families, including
yield and tomato fruit quality negatively. Amaranthaceae, Asteraceae, Chenopodiaceae
Chemical control methods have been trusted to and Poaceae (Siqueira et al., 2000).
control this insect, but the feeding habits of the Other cultivated Solanaceous species host-
larvae, the increasing number of resistant plants are Solanum tuberosum, S. melongena
strains of this pest, together with the negative and Nicotiana tabacum are cited (García and
impact of the chemical into the environment Espul, 1982).
makes the chemical control method not Tuta absoluta was detected in Europe for the
sustainable (Moreno, 2011; Cocco et al., 2012). first time in the northern part of eastern Spain
Thus, the use of nature-friendly biological at the end of 2006. Since then, it has rapidly
control methods and tolerance varieties has invaded other European countries, including
been preferred in recent years. Therefore, the Norway and threatening tomato cultivation
implementation of environmentally safe (Sundheim, 2017), and spread throughout the
measures that reduce the use of chemicals will Mediterranean area, including parts of North
contribute to the sustainability of productions. Africa and the Middle East where it
There are many insects, diseases and weeds immediately reached damaging levels and fruits
which cause significant damage to tomato (Desneux et al., 2010). At present, T. absoluta
production. The most widespread of these are is considered to be one of the most devastating
the whitefly (Bemisia tabaci), leaf miner pests for tomato crops (Solanum lycopersicum
(Liriomyza trifolii), and red spider L.) and a serious threat in all of these newly
(Tetranychus urticae) (Uygun et al., 1998). The infested areas. It is still one of the major pests
431
in many areas of Turkey. As a result, it has pest resistance among plants of this accession,
damaged the tomato production, both in open especially as related to plant chemical
field and greenhouse areas. Tuta absoluta composition, are unknown. Host plant
larvae can absolutely destroy the tomato resistance can be an important component of
canopy by excavating the leaves, stems, and IPM programs. Resistant plants can help
buds and burrows into fruits causing the quality maintain pest populations below economic
decline of fresh tomato and yield loss that injury levels and are usually compatible with
range from 50% to 100% (Cocco et al., 2012). other control methods (Smith, 2005). Sources
According to literature, against T. absoluta of resistance to several insect pests have been
many control strategies have been used such as identified in wild tomato species (Oliveira et
cultural hygiene, pheromone, biological control al., 2012; Bottega et al., 2015).
and resistant genotypes. Nowadays, most of the In this study, we investigated interactions
European countries producers fulfil the among some different tomato genotypes and
GLOBALGAP requirements, which also Tuta absoluta by assessing on number of larvae
implies more environmentally friendly and eggs reared.
technologies. One of the most important ways
of sustainable nature-friendly growth is the MATERIALS AND METHODS
development of disease-tolerant plants. The
plants have shown 3 basic mechanisms against In the experiment, Solanum chmielewskii
insects. One of them is antixenosis. According (LA1028), Solanum neorickii (LA0247),
to this defensive mechanism, the plants cannot Solanum hirsutum (LA1777) S.
be damaged by insects in terms of the plant's pimpinellifolium (LA0722), Solanum
characteristics and cannot feed and lay eggs on lycopersicum cv. Cuatomate, and Solanum
the plants. The second mechanism is antibiosis. lycopersicum cv. Ailsa Craig (LA2838A) were
This feature is a deterioration of the biology of used as plant material. Seeds were germinated
the insect with respect to the characteristics of in a mixture of peat: perlite: vermiculite in
the plants. The third mechanism is tolerant 7:2:1 ratio. After 15 days, tomato seedlings at
plants. This attribute is different from the first the second-true leaf stage were transferred to 8
two, in which case there is no relationship L plastic pods included peat: perlite (2:1)
between plant and insect. As an example of this mixture. The experiment was carried out in a
feature; wild relatives of tomato have been used glass greenhouse, with day temperature 28-
as sources of insect resistance (Oliveira et al., 30oC, night temperature 20-22oC and 60-70%
2009). Insect resistance is generally associated relative humidity in summer time (June-
with the presence of trichome, of different August). Plants were grown up until 8-10 real
types and densities (McDowell et al., 2011; leaf stages for Tuta absulta application in the
Tissier, 2012; Glas et al., 2012). Antibiosis and greenhouse equipped with automatically-
antixenosis are the two resistance mechanisms controlled side walls, ventilation fans and wet-
reported in S. hirsutum (Weston et al., 1989; pads for humidity control and in order to
Channarayappa et al., 1992; Eigenbrode et al., maintain a maximum temperature of 30°C.
1993; Eigenbrode et al., 1994; Kumar et al., The seedling, irrigated with a nutrient solution
1995). Sesquiterpenes present on leaf glandular of EC: 2.2-2.5 and pH: 5.5-6.0 were planted to
trichomes of S. hirsutum f. typical, mainly type 8 L pods after 7 days. The concentrations of the
IV and VI, are commonly reported in the following elements were determined for the
literature as the chemical causes of insect nutrient solution: (M): 3.0x/10-3 Ca(NO3)2;
resistance in this genotype (Snyder and Hyatt, 0.9x/10-3 K2SO4; 1.0x/10-3 MgSO4; 0.2x/10-3
1984; Lin et al., 1987; Carter et al., 1989; KH2PO4; 1.0/10-5 H3BO3; 1.0x10-6 MnSO4;
Eigenbrode et al., 1993; Eigenbrode et al., 1.0x/10-7 CuSO4; 1.0x/10-8 (NH4)6Mo7O24;
1996; Snyder et al., 1998). Although several 1.0x10-6 ZnSO4; 1.0x/10-4 Fe EDTA.
studies have shown that the accession LA1777 When the tomato plants reached 8-10 true leaf
of L. hirsutum f. typicum constitutes a stage, were taken in a 2000 lux illuminated
resistance source to tomato insect-pests, its climate chamber with T. absoluta production
resistance to T. absoluta and the variability of
432
temperature of 25 ± 1°C and 60 ± 5% relative However, in a previous study, the accession
humidity conditions. line LA1777 (S. hirsutum) was found more
The plants were left in this room for 7 days tolerant genotype to Tuta absulta (Ecole et al.,
with T. absoluta. After this period, the plants 1999). To examine the survival ability of the
were moved into the insect-proof climate insect the number of the eggs was determined
chamber under 25°C, 65% relative humidity, for all genotypes. In the first-week egg counts,
and 2000 lux illumination, where the phenotype S. chmielewskii, S. pimpinellifolium, S.
experiments were performed for six weeks. lycopersicum cv. Cuatomate, S. neorickii, S.
Six different genotypes were arranged in a lyocpersicum and S. hirsutum genotypes were
randomized complete design with five found to have average 4.4, 3.0, 3.0, 3.4, 3.2 and
replications. T. absoluta was regenerated one 3.2 eggs respectively on the leaves. In the
times a week and maintained by feeding the second week 0.8, 0.8, 0.8, 0.4, 1.2 and 1.2 eggs
clean tomato plant, S. lycopersicum cv. Ailsa were observed respectively (Table 1).
Craig. Each week 5 leaves were taken from After the second week only S. lycopersicum cv.
each tomato genotype and eggs and larvae were Cuatomate survived, all other genotypes of the
counted at intervals of 7 days. Larvae and eggs green canopy being completely destroyed by
were counted under a Leica® S6D stereo Tuta absoluta. For this reason, subsequent
microscope. Other pictures were taken by counts continued with S. lycopersicum cv.
Canon® D600 digital camera (Figure 1). Cuatomate. The number of eggs during 6
All analyses were performed with SPSS weeks was 3.0, 0.8, 0.2, 0.4, and 0.0, respec-
software package Version 18.0 (SPSS Inc., tively. It was observed that this resistant geno-
Chicago, IL) for Windows by General Linear type died after the 5th week as well (Table 1).
Model univariate test. Normality of variance
was checked before analysis, and the, mean Table 1. Number of the eggs according to tomato
varieties during the six weeks (Sc: S. chmielewskii, Sp:
was compared using an LSD test. S. pimpinellifolium, SlC: S. lycopersicum cv. Cuatomate,
Sn: S. neorickii, Sl: S. lyocpersicum, Sh: S. hirsutum)
Genotypes
Sc Sp SlC Sn Sl Sh
1. Week 4.4 3.0 3.0 3.4 3.2 3.2
Egg Larvae Adult 2. Week 0.8 0.8 0.8 0.4 1.2 1.2
Figure 1. Larvae, egg, and adult of Tuta absoluta 3. Week 0.0 0.0 0.8 0.0 0.0 0.0
4. Week 0.0 0.0 0.2 0.0 0.0 0.0
RESULTS AND DISCUSSIONS 5. Week 0.0 0.0 0.4 0.0 0.0 0.0
6. Week 0.0 0.0 0.0 0.0 0.0 0.0
According to results, LA1028, LA0247,
LA1777, LA0722, and LA2838A were
determined more sensitive than Solanum In the first week of larval counts, S.
lycopersicum cv. Cuatomate to Tuta absoluta chmielewskii, S. pimpinellifolium, S.
(Figure 2). lycopersicum cv. Cuatomate, S. neorickii, S.
lyocpersicum, S. hirsutum genotypes were
observed to have average 6.4, 3.6, 3.6, 2.8, 3.4
and 3.4 larvae, respectively. In the second week
counts, mean 11.6, 6.2, 6.2, 4.4, 5.8 and 5.8
larvae were determined, respectively. After two
weeks, only one tomato genotype (S.
lycopersicum cv. Cuatomate) survived while
other genotypes were found to be completely
dead. On this genotype, the counts in the third,
fourth and fifth weeks, the average number of
larvae was found as 5.4, 5.8 and 7.4,
respectively (Table 2).
Figure 2. S. lycopersicum cv. Cuatomate and other
used genotypes in the study at the 2nd week
433
As a result of both counting of eggs and larvae, However, in the larva counts, low amount of
5 species were severely damaged by the T. larvae were determined on S. lycopersicum cv.
absoluta attack, while S. lycopersicum cv. Cuatomate and it was determined that the
Cuatomate has been found to be more tolerant. difference between the other species was
These results indicate that S. lycopersicum cv. statistically significant (P <0.05).
Cuatomate genotype could be more tolerant to According to other studies, sources of
T. absoluta then others. resistance to several insect pests have been
identified in wild tomato species (Baldin et al.,
Table 2. Number of the larvae according to tomato 2005; Oliveira et al., 2012; Bottega et al.,
varieties during the six weeks (Sc: S. chmielewskii, Sp: 2015), and the corresponding genes have been
S. pimpinellifolium, SlC: S. lycopersicum cv. Cuatomate,
Sn: S. neorickii, Sl: S. lyocpersicum, Sh: S. hirsutum)
introgressed into commercial cultivars.
For example, tomato plants possess glandular
Genotypes
trichomes that accumulate metabolites toxic to
Sc Sp SlC Sn Sl Sh
herbivorous insects (Weinhold and Balwin,
1. Week 6.4 3.6 3.6 2.8 3.4 3.4
2011).
2. Week 11.6 6.2 6.2 4.4 5.8 5.8
3. Week 0.0 0.0 5.4 0.0 0.0 0.0 CONCLUSIONS
4. Week 0.0 0.0 5.8 0.0 0.0 0.0
5. Week 0.0 0.0 7.4 0.0 0.0 0.0 S. lycopersicum cv. Cuatomate tomato variety
6. Week 0.0 0.0 0.0 0.0 0.0 0.0 was determined to be more resistant to T.
absoluta than other varieties. On the other
It is known that besides the reactions of the hand, S. chmielewskii, S. neorickii, S. hirsutum,
wild lines tomato against the tomato leaf miner, S. pimpinellifolium and S. lycopersicum tomato
different kinds of reactions are given within varieties had a high number of
themselves in some commercial varieties as total eggs per female on the first two weeks
well. Indeed, in a study by Çekin and Yaşar (Table 3). We saw that the larvae did not
(2015) they determined the life schedule choose the S. lycopersicum cv. Cuatomate
parameters of T. absoluta on comercial tomato when compared to others. For this reason, it is
varieties of Newton, Caracas, Torry, and thought that this variety can be used by
Simsek. breeders for its resistance to Tuta absoluta.
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436
RESPONSES ABOUT SEED FORMATION

AND SEED PRODUCTION IN DIFFERENT GENOTYPES
OF CAPE GOOSEBERRY (PHYSALIS PERUVIANA L.)
Nikolay PANAYOTOV
Abstract
The main goal of the present study was to be established the responses in seed formation and seed production in several
genotypes of cape gooseberry (Physalis peruviana L.) with a target to predict the seed yield and its realization. The
experiments were carried out in Agricultural University of Plovdiv, Bulgaria with six genotypes of the different origin.
The number of seed per fruit, the percentage of normally developed seed, the weight of 1000 seeds, germination energy,
germination, mean germination time, uniformity of germination, length of embryo root and hypocotyls and fresh weight
and deviation in seedlings were established. The weight of seeds per kilogram fruit, the yield of fruit and seed also have
been determinated. Significant variation in the numbers of seed per fruit was observed. High germination was an
account of each genotype. The differences in seed production and sowing quality between genotypes were registered.
These results can be applied for the prediction of seed productivity and also in the determination of the price of seed
depending on the genotyping insemination characteristics.
Key words: germination, seedlings, fruit, yield, germination time.
INTRODUCTION ixocarpa Brat. and Physalis pubescens L.

(Moriconi et al., 1990; Crawford, 2004), the
According to Geleta et al. (2005), the genetic cape gooseberry (Physalis peruviana L.) being
diversity is very important in relation to the most widely used. Christov (2010) is of the
increase the peculiarities of plant material in opinion that production and trade with cape
both scopes, to improve the production as well gooseberry have increased over the last
as use in breeding. The authors emphasized that decades. Its cultivation in the countries of
the presence of more genotypes is considered Europe has become more perspective,
as one of the criteria for the selection and for particularly appropriate for warmer climates
the reaching of the opportunities of the and for small-scale farms (Paksi et al., 2007;
breeding programs. The importance and Popova et al., 2010). According to this author
availability of rich genetic material of various (Christov, 2010), varieties as a product of
origins, which are suitable to be included in targeted selection are practically absent and
selection activities are reported also from local forms and ecotypes are being grown
Martirosyan and Sargsyan (2014). massively, therefore it is necessary to
Many researches established strong genotypic investigate different genotypes and ecotypes
response, studied a wide range of varieties in under different conditions.
various vegetable crops (Haytova and Experiments with a large number of genotypes
Babricov, 2006; Haytova and Gergova, 2011; of cape gooseberry outdoors and under
Todorova 2011; Todorova et al., 2011; controlled conditions are conducted by Abak et
Todorova and Pevicharova, 2012) al (1994) in Chukurova, Turkey. Panayotov
The genus Physalis covers on significant (2010) reports on the selection of the first
number of annual and perennial species, variety of cape gooseberry in Bulgaria, called
according to various authors between 70 Plovdiv.
(Christov, 2010) and 110 (Skvorcova, 1997). This researcher examines the adaptability of
The most common and practical applications genotypes of Physalis peruviana L. with
are three main species, namely Physalis different origins, with a goal directed to
peruviana L., Physalis pruinosa L., Physalis identifying the most suitable for cultivation in
437
the region of Southern Bulgaria (Panayotov, University of Plovdiv, Bulgaria with several
2016). McCain (1993) and Crawford (2004) genotypes cape gooseberry (Physalis peruviana
reported that cape gooseberry is propagated L.) with different origin:
most commonly by seeds. Chernok (1997) 1. Variety Plovdiv - origin from Bulgaria;
confirms the thesis that this crop is grown by 2. Genotype Obrazec 1 - origin from the USA;
seeds, pointing out that its cultivation very 3. Genotype Obrazec 2 - origin from the USA;
often takes place by direct sowing on the open 4. Genotype Obrazec 3 - origin from the USA;
fields. This requires to be provided a sufficient 5. Genotype 08-2010 - origin from Turkey;
quantity of high-quality seeds. 6. Genotype 11-2012 - origin from Germany.
Therefore the scientific researches into the The plants were grown by unpricked seedlings,
process of seed production are necessary and produced in a plastic unheated greenhouse with
must be carried out. In terms of seed production sowing on March 15th. The seed rate per one
of the cape gooseberry, the published studies in hectare was 80-100 g, and per square meter was
the world literature are insufficient and very sown at 1.5 g. The planting took place between
few. Lawrence (1993) points out that seed May 20-25 on furrows, on the scheme 70 x 50
production of cape gooseberry is related to cm (Panayotov and Tcorlianis, 2000). All nece-
genotypic belonging, on the other hand, ssary agro-technological practices were applied
depends on the size of plants, which affects the during vegetation. The experiments took place
quantity and vitality of the pollen and hence the in four replicates with 25 plants in each and the
seed productivity. size of the experimental plot was 9 m2.
In the larger plants, sometimes the amount of At the stage of full botanical maturity, all the
pollen is reduced, but because they develop ripe fruits were harvested and their yield was
more flowers, their number of seeds is higher, determined. The seed was extracted and the
in opposite to smaller ones. In order to increase seed yield was established. The seed yield from
seed production, the author recommends one kilogram of fruit was studied. Separately
additionally applying manual pollination to on 5 fruits from each replicates all seeds were
support the natural ones. extracted.
The effect of this manipulation is much higher The following indicators: number of seeds per
in genotypes that develop weaker plants, as fruit - determined by counting the seeds of five
they also rely on a small number of flowers. fruits of each replicates; percentage of fully
Thomson and Wit (1987) investigated the developed seeds in one fruit (insemination) - by
optimal temperature regime for both the growth counting of the normally developed seeds and
and the germination of the seeds of two types calculating their percentage to the total number
of physalis - Physalis angulata L. and Physalis of seeds per fruit were registered.
virginiana L. var. underglared. They establish The linear seed sizes - length, width and
a strong species and genotype response, both in thickness of seed, measured on 15 seeds with
seed set up and seed development, as well as electronic calliper; weight of one seed - set on
the temperature regime for maximum 15 seeds, weight of 1000 seeds (ISTA, 2013),
sprouting. in 4 replicates; germinating energy (ISTA,
The main goal of the present study was to 2013), in 4 replicates; germination (ISTA,
achieve a comparative evaluation of develop- 2013) in 4 replicates; uniformity of germination
mental features, productivity and vitality status (according to Strona, 1966); mean germination
of the seeds of different genotypes of cape time (MGT) - in four replicates, each of 100
gooseberry (Physalis peruviana L.) in relation seeds, calculated according to the equation
to predicting of the seed yield and the need for given below were determinated.
a specific approach for its realization. Σ (G x T)
M.G.T. = , where:
MATERIALS AND METHODS F
T - day in which the seed is germinated;
The experiments were carried out in 2015-2017 G - numbers of seeds that germinated in this;
years in the Scientific field and laboratories of F - final number of germinated seeds (Battle
Department of Horticulture at the Agricultural and Whittington, 1969).
438
On the day of counting of germination in, the genotypes is even greater, almost double (64.2)
four replicates: the fresh mass of a seedlings, numbers. The differences between the variants
by measuring all developed seedlings; length of are of statistical significance with exception of
the embryo root and of the hypocotyl - by those between Obrazec 2 and Obrazec 3 and
measuring 10 seedlings from each replicates; between Ovbrazec 3 and line 08-2010.
deviations from the normal structure of the Besides inseminating of the fruit, the quantity
seedlings by Welington (1970) - short embryo of fully developed seeds is of particular
root, lack of branches in embryo root, lack of importance. In almost all tested cape
hairs on the embryo root, undeveloped gooseberry genotypes, most seeds are fully
cotyledons, unopened cotyledons, lack of developed. For Plovdiv variety and Obrazec 3,
hypocotyl were established. they reached up to 100.0%.
Data of the study were subjected to analysis of Despite the highest number of seeds found in
variance, and the least significant differences the fruit of line 11-2012, they demonstrated the
between means were calculated by the Fisher least development (93.6%) of the total seed set
test at p = 0.05. A method for ANOVA and a up, reached to the stage of full formation. Both
method for dispersion are described in Fowel the setup and development of seed are directly
and Cohen (1992). Due to the one-way trend of related to the pollination and fertilization
the results of all vegetation, the data presented processes. Similarly, opinion has Lawrence
are average values in the course of three years. (1993), who found differences in seed
production of the cape gooseberry that
RESULTS AND DISCUSSIONS associated with both genotypic belonging and
plant development and pollination.
In the number of seeds setting up in one fruit The weight of 1000 seeds is one of the
the genotypic differences were established important indicators for determining their
(Table 1). The variation on this index is sowing qualities. The average value for all
significant, as results ranging from 61.5 for included in this experimental genotypes was
Obrazec 1 to 145.7 for genotype 11-2012. 1.06 g, and the variations in this indicator were
Relatively smaller numbers of seed there were relatively low. With the highest mass of 1000
developed in variety Plovdiv (73.0) and in air dry seeds is characterized Plovdiv variety
selecting line 08-2010 (88.5) numbers. The (1.14 g), followed by Obrazec 3 (1.11 g). The
average numbers of seeds per fruit for all tested lowest, although with a small difference was
genotypes is 90.7. The variation against this the weight of 1000 seeds of line 11-2012 (1.0
mean value between genotypes with the lowest g), where was mentioned above that the highest
number of seeds and the highest one is between number of seeds in one fruit was reported.
-29.2 and +35. The difference between the There were also shown to be of a low value in
highest and the lowest, established result, for the seed of Obrazec 1 (1.03 g).
the number of seeds in the experimental
Table 1. Cape gooseberry seed formation
№ Genotype Seed % of normal Weight of

number/fruit developed 1000 seeds
seeds (g)
1 Plovdiv 73.0 100.0 1.14
2 Obrazec 1 61.5 97.7 1.03
3 Obrazec 2 99.8 97.8 1.05
4 Obrazec 3 95.9 100.0 1.11
5 08-2010 88.5 95.9 1.04
6 11-2012 125.7 93.6 1.00
LSD p=0.05% 12.2 0.4
439
Besides, as a seed material, according to Zhang observed for the width of the seed from 1.0 mm
and Wen (1996), the cape gooseberry seeds can to 1.4 mm, as well as the thickness from 0.45
be used as an indicator for taxonomic mm to 0.5 mm. Significant, though slight,
identification between species and genotypes of differences are observed at the length of the
the genus Physalis. In the experiments seeds. On average, for the explored sample of
conducted, however, significant differences in genotypes, the length of one seed is 1.92 mm.
seed morphology were not established (Table Deviations from this mean value are from -
2). The weight of one seed was changing to a 0.21 for Obrazec 3 to + 0.23 for genotype 08-
narrow range from 1.15 mg (Obrazec 2) to 1.28 2010.
mg (Obrazec 1). A similar tendency was also
Table 2. Morphology behaviours of cape gooseberry one seeds
№ Genotype Weight (mg) Length Width Thickness

(mm) (mm) (mm)
1 Plovdiv 1.23 1.99 1.2 0.5
2 Obrazec 1 1.28 1.84 1.4 0.5
3 Obrazec 2 1.15 1.86 1.2 0.45
4 Obrazec 3 1.25 1.71 1.1 0.45
5 08-2010 1.21 2.15 1.3 0.45
6 11-2012 1.17 1.99 1.0 0.45
LSD p=0.05% 0.2 0.31 0.2 0.18
The most important indicator for sowing established. The statistical significance between
qualities and for seed development is their Obrazec 1 and Obrazec 2 was not observed.
vitality (Table 3). From genotypes included in In connection with the more prices study of the
the experiment, it can be pointed out that they seed qualities, the indicator mean germination
all have very high germinating energy between time and uniformity of germination (Table 4)
74.0% for genotypes 11-2012 to 98.7% for are of great importance. The mean germination
Plovdiv variety. On this background, the time was the highest in Plovdiv variety (4.38
germinating energy for Obrazec 2 and Obrazec days), followed by the seed on genotype 08-
3 was also relatively high 95.3% and 96.7%, 2010 (4.44 days). The seeds of Obrazec 2
respectively. As Balck et al. (2008) and germinated most slowly, for 5.52 days. This
Copeland and McDonalds (2001) emphasized indicates that the mean germination time is
the seeds that have demonstrated higher relatively high. The uniformity of germination
germinating energy are with better vital range from 23.69% for line 08-2010 to 32.89%
performance because they have germinated for at 11-2012. High uniformity was established in
a shorter period. the Plovdiv variety and also in Obrazec 2 over
Thomson and Wit (1987) reported that the seed 29%. This data once again confirm the opinion
of the cape gooseberry is characterized by high that the cape gooseberry seeds has a good
germination, and yet at the eighth week after vitality and ability to germinate and that their
flowering, it reaches over 90%. In this high uniformity, as Panayotov (2015) and
experiments the germination does not differ Black et al. (2008) maintains, it will allow them
significantly from the values of the germination to better overcome the resistance of soil during
energy, indicating that most seeds are with high sprouting.
vitality. An exception was observed for the The morphological characteristics of the
seeds of line 11-2012, where the germination seedling (Table 5) contribute to a more detailed
exceeds more the data for energy with 12.7%. clarification of the seed vitality status. In the
The highest germination was recorded for the fresh weight of the seedling of one seed, the
seeds of the Plovdiv variety and for Obrazec 1 differences are more significant.
(98.7%), and the lowest one at 11-2012
(86.7%). Mathematical proof of differences is
440
Table 3. Germination behaviours The weight of the seedling indicates the
of cape gooseberry seeds possibilities of seed to germinate more easily
№ Genotype Germination Germination
energy (%) (%)
and is very often used to determine their vigour
1 Plovdiv 98.7 98.7 (Copeland and Mc Donalds, 2001; Panayotov,
2 Obrazec 1 93.3 96.0 2015).
3 Obrazec 2 95.3 96.0 The values range from 14.8 mg for Obrazec 1
4 Obrazec 3 96.7 98.7 and line 08-2010 to 18.3 mg for Obrazec 2.
5 08-2010 93.3 96.7 The length of the embryo root for the seed of
6 11-2012 74.0 86.7
line 11-2012 is the highest and it’s reaches to
LSD p=0.05% 2.6 2.2
3.92 cm. Next one is those of Obrazec 3 with
Table 4. Sowing parameters of cape gooseberry seeds 3.41 cm.
Slightly developed was the embryo root of the
№ Genotype MGT Uniformity line 08-2010 (2.89 cm).
(days) (%) The seedlings of Obrazec 2, Obrazec 1 and 08-
1 Plovdiv 4.38 29.78
2010 are characterized by the highest length of
2 Obrazec 1 4.85 24.0
3 Obrazec 2 5.52 29.33 the hypocotyl 3.91 cm, 3.84 cm, and 3.78 cm,
4 Obrazec 3 4.66 24.67 respectively.
5 08-2010 4.44 23.69 The smallest one was in line 11-2012 (3.4 cm).
6 11-2012 4.64 32.89 These results unambiguously show that the
LSD p=0.05% 1.1 6.2 variation between individual genotypes is
weak.
Table 5. Morphological characteristics of cape gooseberry seedling
№ Genotype Fresh weight (mg) Length of hypocotyl (cm) Length of embryo root (cm)
1 Plovdiv 17.6 3.69 3.15
2 Obrazec 1 16.9 3.84 3.31
3 Obrazec 2 18.3 3.91 3.34
4 Obrazec 3 14.8 3.47 3.41
5 08-2010 14.8 3.78 2.89
6 11-2012 15.2 3.4 3.92
LSD p=0.05% 4.2 2.6 1.9
It is normally some of the seedlings to have for 11-2012 to 52.0% for Plovdiv, calculated
different deviations from their normal towards all not well-developed seedlings which
morphology (Table 6). The percentage of were accepted for 100%. The lack of branching
abnormally seedling grown was the highest in in the root, take the next place, most often
Obrazec 2 (15.6%), followed by line 11-2012 observed in line 08-2010 to (46.6%) and in the
(12.3%). The lowest values were recorded for Plovdiv variety (36.6%). Minor deviations are
the Plovdiv variety. The most common types of associated with lack of hair on the hypocotyl,
deviation are the lack of root hair, from 22.3% unopened cotyledons, and short embryo root.
Table 6. Deviation of normal developed seedlings (%)
Genotypes Short Lack of Lack of heirs Undeveloped Unopened Lack of hairs

% embryo branches on on embryo root cotyledons cotyledons on hypocotyl
root embryo root
Plovdiv 10.5 0.0 36.6 52.0 0.0 11.4 0.0
Obrazec 1 13.2 17.3 24.8 31.4 0.0 15.2 11.3
Obrazec 2 15.6 9.66 15.5 36.6 22.0 15.6 0.0
Obrazec 3 11.2 9.30 10.5 30.3 3.30 46.6 0.0
08-2010 10.8 0.0 46.6 33.0 7.66 12.3 0.0
11-2012 12.3 0.0 35.0 22.3 4.20 38.5 0.0
441
For agronomic and economic evaluation in genotypic characteristics. For samples with a
relation to prediction of the expected seed yield lower seed yield and a higher ratio between the
the seed quantity in one kilogram of fruits in quantities of fruits and the seeds obtained
botanical maturity was determined, and it with therefrom the price is appropriate to be higher,
an average of 30.69 g (Table 7) for the studied because the input costs for fruit production are
genotypes. Most of the seeds of one kilogram almost the same between separate genotypes.
of fruit were obtained in Obrazec 3 (37.95 g), Furthermore, for those with low seed yields,
and in the Plovdiv variety (34.85 g). Relatively there are additional costs associated with
high results were observed for line 08-2010 extraction of more fruits to be obtained the
(31.52 g). The least one in their quantity in one corresponding amount of seed.
kilogram of the fruit was of Obrazec 1 (20.66
g). The statistical significance of the Table 7. Productivity behaviours of cape gooseberry
differences between the tested genotypes is genotypes
established with exception of that between № Genotype Seed yield/ Ratio fruit :
Plovdiv and Obrazec 3. 1 kg fruits (g) seeds
1 Plovdiv 34.85 28.69
The ratio between the quantity of fruit and the
2 Obrazec 1 20.66 48.40
seeds extracted from them contributes to a 3 Obrazec 2 30.79 32.47
complete assessment of the insemination and 4 Obrazec 3 37.95 26.35
indicates how many fruits are necessary to be 5 08-2010 31.52 31.72
extracted in order to be obtained an adequate 6 11-2012 28.37 35.24
quantity of seeds. A high genotypic response LSD = 0.05% 4.4
has been observed since the values of this
Table 8. Productivity of cape gooseberry
marker are in a very large range from 26.35 for
Obrazec 3 to 48.40 for Obrazec 1, or an № Genotype Yiled of fruit Yiled of seed
average for the tested population is 33.85. (kg ha-1) (kg ha-1)
The seed yield of the cape gooseberry is 1 Plovdiv 2245.5 78.2
directly related to the yield of fruit (Table 8). 2 Obrazec 1 1668.3 34.4
The highest yield was harvested in the Plovdiv 3 Obrazec 2 2055.0 63.2
4 Obrazec 3 1581.8 59.9
variety, it is 2245.5 kg ha-1, followed by 5 08-2010 1642.2 51.8
Obrazec 2 (2055.0 kg ha-1). For other 6 11-2012 1599.1 45.6
genotypes it is between 1581.8 kg ha-1 to LSD p=0.05% 120.1 14.0
1668.3 kg ha-1 for Obrazec 3 and for Obrazec 1,
respectively. CONCLUSIONS
The genotypic features of cape gooseberry also
occur with respect to the obtained seeds. Most Genotypic differences were found in the
seeds are obtained from Plovdiv variety with number of seeds in one fruit of cape
78.2 kg ha-1, followed by Obrazec 2 with 63.2 gooseberry.
kg ha-1. This mainly could be due to the higher The percentage of fully developed seeds for all
yield of fruits and on the other hand to the cape gooseberry lines is high. Slight differences
relatively low ratio fruits: seeds. The least are noted with respect to the morphological
yield, 34.4 kg ha-1, was recorded for Obrazec 1, features of the seed.
the genotype with the lowest fruit productivity The seeds of the cape gooseberry are
and a quite high ratio of fruit: seed weight. The characterized by high vitality.
average seed yield of the tested genotype of More significant are the differences in the fresh
cape gooseberry was 55.51 kg ha-1. The weigh of the one seedling. The type of
differences according to seed productivity deviations from the normal development of
between Plovdiv and Obrazec 1 and Obrazec 2 seedling structure most often they included the
and also between Obrazec 1, and Obrazec 2 and lack of hairs or branchings of the embryo root.
Obrazec 3 are mathematically proven. Genotypic differences also exist in
These results can be used to predict seed insemination. The fruit-to-seed ratio also
productivity and, at the same time, to determine changes in a wide range.
the selling price of seeds, depending on
442
The average seed yield between tested research on the Union of scientists in Bulgaria-
genotypes of cape gooseberry is 55.51 kg ha-1, Plovdiv, series C. Techniques and Technologies,
Union of Scientists Session 11-12 November, VІІІ:
and variations between individual genotypes 196-199 (Bg).
being are significant. International Rules for Seed Testing, 2013. International
The obtained results can be used on one hand to Seed Testing Association. Bassersdorf, CH-
predict the expected yield of cape gooseberry Switzerland.
seeds and on the other hand to precise the Lawrence W., 1993. Resource and Pollen Limitation:
Plant Size - Dependent Reproductive Patterns in
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genotyping insemination characteristics. initial material for pepper breeding. Acta
Horticulturae, 1033: 47-52.
McCain R., 1993. Golden berry, passion fruit, & white
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support of Scientific Project 15-15 of Moriconi D., Rush M., Flores H., 1990. Tomatillo: A
Agricultural University-Plovdiv, Bulgaria, potential vegetable crops for Louisiana. In: Janick J.,
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NEW BRED CULTIVARS OF BELL PEPPER

OBTAINED AT V.R.D.S. BUZĂU
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1
Vegetable Research-Development Station Buzău, 23 Mesteacănului Street, Buzău, Romania
2
Faculty of Horticulture, 59 Mărăști Blvd, Bucharest, Romania
Abstract
Among the vegetables grown in Romania, bell peppers occupies a priority place due to favourable pedo-climate
conditions, especially in the southern area of the country, as well as of high demand from consumers and food-
processor. In Romania, peppers has traditionally been field-grown and harvested at mature-green stage. Lately, as a
result of worldwide producers, the expressiveness of colour and shape had greatly diversified and the demand for high-
quality coloured peppers has led farmers to look at quality-pepper cultivars grown in greenhouse. Starting from these
premises, the Breeding Laboratory at Vegetable Research-Development Station Buzău approach a new theme aimed to
obtain new cultivars with high yield potential and distinct phenotypic characteristics. Besides evaluation of yield
potential, a particular emphasis has been focused on obtaining varieties that has different shapes and colours.
Research has been completed by far with the achievement of new cultivars with colour ful fruits like orange, chocolate
and indigo.
Key words: breeding, Capsicum annuum L., Romania, coloured pepper.
INTRODUCTION phytochemicals (Chuah et al., 2008)

According to a study, the colour of bell peppers
In the past, in Romania, farmers limited is the major factor associated with the
themselves and had cultivated only traditional consumer purchasing decisions (Frank et al.,
varieties of peppers, but now, as a result of 2001). Starting from these premises, Breeding
imported seeds and imports of different kind of Laboratory from Vegetable Research-
peppers, the expressiveness of colour and shape Development Station Buzau, had approached a
has greatly diversified. new theme aimed to obtain new genotypes with
Pepper (Capsicum annuum L.) is an important high yield potential and distinct phenotypic
vegetable crop worldwide and can be characteristic. So far, V.R.D.S. Buzau has a
consumed in many colours green, red, yellow, valuable collection of germplasm of peppers.
orange, or brown when ripe (Ilić et al., 2014). Acquaah (2012) states that germplasm needs to
Improvements made in pepper breeding sector be periodically rejuvenated and multiplied,
have allowed a diverse and large number of because it represents the life of plant breeding
varieties into the market with distinct and without it, this activity could not be carried
expressivity in accordance with direction of out. Thus, it is used a genetic material for
use. Peppers can be used fresh in salads, breeding and can itself be improved in order to
cooked in different dishes, used as a hot or increase the performance of the biological
sweet spice, and also in the industry, for pickles material obtained.
and different kind of preserves. They are fast Any breeding process begins by collecting the
gaining popularity, not only for their attractive genetically diversified germplasm that must
colours, characteristic taste and aroma, but also incorporate a sum of features useful for the
for their health-promoting properties. objectives pursued. The results obtained are
Peppers exhibit great genetic diversity in terms determined by a large extent and abundance of
of colour, size, shape and chemical genetic sources, by the combination of useful
composition and therefore vary greatly in their genes of which accumulation is sought in the
antioxidant properties, vitamins and other organisms created. (Leonte C.N., 2011).
445
In this study, a special emphasis was placed on RESULTS AND DISCUSSIONS
obtaining new genotypes of bell peppers with
different shapes and colours, compared to the Sowing was made on 7th March 2017 and after
known ones, while evaluating their yield 12 days A 60 was the first one who sprout, with
potential. a germination percentage of 88%, it was
One important part in the breeding sector is the followed by A 68 with the highest germination
evaluation of the germplasm collection and the rate, 95%, afterwards A 62 had the lowest
comparison between a few superior candidate germination percentage only 81% and the last
genotypes to select one or more to be released one who sprout was after 17 days A 66 with a
as cultivars. The goal of this research was to germination rate of 91%.
evaluate four ascensions and to study their Laboratory germination was also made and in
potential to identify the most promising one. Figure 1 can be observed the difference
between the two of them.
100
96
Researchers who are working in the breeding 95
95
sector have to choose between hundreds of 92
89
90 Percent
plants in a segregation population to select only 90 88
germination
a small fraction of promising plant to be used in 85
84
laboratory test
81
the advanced program. So far, V.R.D.S. Buzau 80 Percent
has ended with the achievement of a valuable germination
germplasm collection and has a number of 214 75 greenhouse
Capsicum genotypes, structured in three groups 70
regarding genetic stability: segregating, A 60 A 62 A 66 A 68
advanced and stabile.
From a number of 42 varieties of Capsicum Figure 1. Germination results
annuum var. grossum L., there have been
selected 4 ascensions (A 60, A 62, A 66 and A Overall, at the beginning, the seeds germinated
68) with distinct phenotypic characteristics. more slowly in the greenhouse than the lab.
Afterwards they have been genetically The highest difference was recorded by A 60,
stabilized by specific, long term breeding work. with a decrease of 4% towards laboratory
On 7th March the seeds were sown in seedling germination value.
trays with 70 cells, in a substrate with white The ascensions A 62, A68 were also followed
peat and added nutrients. In the 3rd of May they by a decrease towards laboratory germination
were transplanted in a greenhouse. value. The germination conditions in the lab are
The specific care work was respecting the crop very controlled compared to the greenhouse;
technology and it was added the special this may explain why there were some
techniques that consisted in handling the bell differences between germination results.
pepper shoots and the removal of the first floral Only A 66 has recorded a 1% higher than
button. laboratory germination analyses, but it has
For reducing the damage caused by high sprout at last. The peculiarities of climatic
temperatures and sunburn, starting from 15th of zoning for Buzau in 2017, was that in the
June, a shading net had been mounted on the second decade of April, there have been snow
top of the greenhouse. This had led to a and ice frosts, for why vegetable crops had
decrease of damage caused by flower abortion delayed till 15 days.
and sunburn fruits. In this study, there were Planting was made in greenhouse on 3rd May
analyzed one hundred plants for each 2017, when the seedling has reached the
experimental variant. optimum planting age and the climatic
During the vegetation period, phenological and conditions were favourable.
biometric observations were made according to Plant growth habit on Ascension 60 (A 60,
IPGRI standards for Capsicum spp. Figure 2) was intermediate and has recorded an
average height of 85 cm (stage 4 in IPGRI
descriptors).
446
colour when ripe. As an important notice, it has
been found that fruit tend to lose their flavour
on their ripening process. The root system has a
medium vigour and a medium to weak resis-
tance to pest. There was a slight attack of ne-
matodes on the roots (Meloidogyne spp.), but
did not influence in a significant percentage the
yield.
Figure 2. Ascension 60, plant view
Ascension 66 (A 66, Figure 4) has recorded the
lowest average height from studied ascension
Bell pepper fruit set percent was high, even on and fits to the lower limit of IPGRI descriptors
high temperature during summer. The imam- (75 cm).
ture fruit is dark, indigo-black, crispy, aromatic
and can be consumed fresh or cooked. At
physiological stage fruits are red. When fruits
turn red they have a sweet and strong aromatic
flavour. More details about fruit characteristics
can be found in Table 1. The root system is
medium developed and shows medium
resistance to pests (Meloidogyne spp.).
Table 1. Fruits characteristics

Studied character A 60 A 62 A 66 A 68
Fruit colour at Indigo- Dark Intense Green
intermediate stage black green indigo-black
Fruit colour at mature Red Orange Dark red Brown
stage
Figure 4. A 66, plant view
Fruit average length 10.62 8.71 7.99 11.68
(cm)
Fruit average width 10.12 8.53 9.23 7.83
Observations made have shown that this ascen-
(cm) sion grows well and fruit set in vary conditions.
Fruit average wall 9.83 6.93 6.10 6.45
thickness (mm) It has recorded very good results on high
Fruit average weight 158.1 142.1 133.5 130.03 temperature during summer and also on low
(g)
Fruit average pulp 124.8 142.1 114.9 119.73 temperature during autumn (October). Interme-
weight (g) diate fruits are darker in colour than A 60 and
Fruit average recep- 33.3 23.2 18.6 10.3
tacle weight (g) at physiological stage turn dark red. The fruit
taste is good, full of flavour in both ripening
Ascension 62 (A 62, Figure 3) has a interme- stages. The root system is vigorous and shows
diate plant growth habit, with an average height medium resistance to pest (Meloidogyne spp.).
over 92 cm (stage 5 in IPGRI descriptors). Plant growth of Ascension 68 (A 68, Figure 5)
was intermediate, with a average height over 85
cm, according to IPGRI descriptor in 5th stage.
Figure 3. A 62, plant view
Fruit set was medium during hot summer.

Unripe fruit has a dark-green colour and orange Figure 5. A 68, plant view
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Fruit set is medium during high temperature in with 1.677 kg/plant. Fruits of smaller size and
the summer. On intermediate fruit stage, the with minor flaws had been included in the
fruits are green and, as they ripen, they turn second class and the registered average yield
brown. Fruits have gone pretty fast to brown varies from genotype to genotype. The highest
colour. The fruits had been remarkable by its yield was recorded by A 62, with a value of
taste and aroma. Root system is medium 0.98 kg/plant and the lowest yield was reported
developed and has a medium pest resistance by A 60 with a average yield of 0.9 kg/plant.
(Meloidogyne spp.). As a result of long vegetation period of bell
In figure 6 are presented fruits in different pepper in Romania, it was found that all studied
ripening stages. ascensions have a high yield potential, but
some of them do not reach full ripening. In this
study it has been framed as a 3rd class yield, a
yield that cannot be marketed due to unripening
of the fruits and their smaller size. For all
ascensions, fruits bellow 100g were included
on the 3rd class. The highest yield for the 3rd
class was registered by A 66 with a value of
1.19 kg/plant and the lowest yield was recorded
by L 62 with only 0,499 kg/plant. More details
about yields registered by other ascensions can
Figure 6. Fruits in different ripening stages
be found in Table 2. Recording a large number
of fruits from the 3rd class has led to the
Regarding yield potential, an assessment of conclusion that the varieties can also be grown
quality and quantity of yield was made for each in a prolonged cycle in a heated greenhouse
studied genotype. In relation to yield quality, and in this case, the yield of first and second
the fruits were separated in: first, second and class will have a significant increase.
third class. Records of obtained yields of all Regarding total yield potential of ascensions
ascension are found in Table 2. the record was owned by A 66 with a value of
4.32 kg/plant and the lowest yield was reported
Tabel 2. Obtained yield by A 62 with a 3.16 kg/plant.
Ascensions A 60 A 62 A 66 A 68
Fruit average 224.4 186.4 180.7 179.1
weight, 1st class CONCLUSIONS
(g)
No fruits 1st class 8 9 12 10
Average Yield of 1779.2 1677.6 2168.4 1791 The absence of national cultivars of bell pepper
1st class/plant (g) with different colour adapted to Romanian
Fruit average 150 140 120 120
weight 2nd class
pedo-climate conditions motivated the
(g) beginning of this research at V.R.D.S. Buzau.
No fruits 2nd 6 7 8 8
class
In the study, four ascensions were examined
Average Yield of 900 980 960 960 with distinct phenotypic characteristics which
2nd class/plant (g) will be enrolled and tested at ISTIS to be
Fruit average < 100 < 100 < 100 < 100
weight 3rd class patented. Ascension 68 had distinguished by a
(g) special colour with a distinctive flavour and a
No fruits 3rd class 6 5 12 7
Average Yield of 599.4 499.5 1198.8 699.3 potential yield plant of 3.45 kg/plant. It is well-
3rd class/plant (g) known that bell pepper fruit set is very
Yield/plant (g) 3278.6 3157.1 4327.2 3450.3
Yield/plant (kg) 3.28 3.16 4.32 3.45 sensitive to environmental condition, in
particular, to low or high temperatures that can
First-class fruits had the organoleptic affect pollen development and another
characteristics of the variety, were uniform in dehiscence. From all studied genotypes,
colour, taste, appearance, texture and size. Ascension 66 has recorded the highest fruit set
Ascension 66, has recorded a medium yield in extreme conditions and the highest yield,
potential with a value of 2.168 kg/plant, and the 4.32 kg/plant. Ascesion 60 was noted with
smallest one was registered by Ascension 62 earlier fruits and a yield of 3.28 kg/plant. The
448
lowest yield potential was recorded by Frank C.A., Nelson R.G., Simonne E.H., Behe B.K.,
Ascension 62, with only 3.16 kg/plant. Simonne A.H., 2001. Consumer preferences for color,
price, and vitamin C content of bell peppers.
Hortscience 36: 795-800.
REFERENCES Ilić S.Z., Šuni L.Ć, Mirecki N., Fallik E., 2014. Cultivars
differences in keeping quality and bioactive
Acquaah G., 2012. Principles of plant genetics and constituents of bell pepper fruit during prolonged
breeding. John Wiley & Sons. storage. J. Adv. Biotech .4: 313-318.
Chuah A.M., Lee Y.C., Yamaguchi T., Takamura H., IPGRI, 1995. Descriptors for Capsicum (Capsicum spp.).
Yin L.J., Matoba T., 2008. Effect of cooking on the International Plant Genetic Resources Institute, Rome.
antioxidant properties of coloured peppers. Food Leonte C.N., 2011. Tratat de ameliorarea plantelor.
Chemistry, 111 (1), 20-28 Editura Academiei Române, București (219-220).
449
450
PRELIMINARY STUDY RELATED ON YIELD

AND QUALITY POTENTIAL OF TWO NEW SWEET PEPPERS
VARIETIES OBTAINED AT V.R.D.S BUZĂU
Elena BARCANU-TUDOR1, Elena Maria DRĂGHICI2, Aurora DOBRIN3, Andreea STAN3,

Carmen CONSTANTIN3, Mihaela ZUGRAVU3, Oana BUJOR-NENIȚĂ3
1
Vegetable Research-Development Station Buzău, 23 Mesteacănului Street, Buzău, Romania
2
59 Mărăști Blvd, Bucharest, Romania
3
University of Agronomic Sciences and Veterinary Medicine of Bucharest, Research Center for
Studies of Food Quality and Agricultural Products, 59 Mărăști Blvd, Bucharest, Romania
Abstract
Sweet pepper is an important vegetable used in Romania and it holds a significant area crop. At present, there are a lot
of new varieties of sweet peppers in Romania, but, unfortunately, most of them comes from worldwide seed producers.
Being demand for autochthonous competitive varieties, suitable for growing in greenhouse, Vegetable Research-
Development Station Buzău has been preoccupied with obtaining of new varieties of sweet peppers. This study was
conducted to investigate the qualitative and quantitative potential of two new varieties, ˈIdealˈ and ˈCarminˈ. As the
study shown, ˈIdealˈ variety has a higher yield potential, while ˈCarminˈ variety has a higher content of vitamin C. Dry
matter content and soluble substances it has higher values in ˈIdealˈ variety. New bred varieties have demonstrated
downstream genetic stability in the phenotypic expressiveness of the main characters and have also recorded high-
quality yield production, thereby contribute to the enrichment of the inland assortment suitable to be cultivated in
greenhouses.
Key words: breeding, Capsicum annuum L., Carmin, Ideal, Romania.
INTRODUCTION 2007). These characteristics are still important

in defining the shape of these fruits and it’s
Capsicum annuum L. var. grossum Sendt also fundamental to determine the group to which
known as bell pepper, sweet pepper and green they belong whether conical, rectangular or
pepper is one of the well known and productive square (Ramos et al., 2017). ˈCarminˈ fruits
species from Solanaceae family and holds belong to the square group and ˈIdealˈ has fruit
significant crop areas in Romania. An with conical shape both of them are from
important part of Romanian varieties of Capsicum annuum var. grossum. It is well
Capsicum spp. are not suitable for growing in known that pepper fruits are very used and
the greenhouse, that is why one of the main appreciated by their high content of ascorbic
objectives of Vegetable Research-Development acid (AsA) and antioxidant activities (carotene
Station (V.R.D.S.) Buzău, was to obtain and flavonoids) (Burzo, 2015). AsA, the anti-
valuable genotypes. V.R.D.S. Buzău owns a scorbutic factor, is a required human nutrient,
diverse germplasm collection of different kind and its biological functions are centred on its
of peppers. Recently, two varieties of peppers, antioxidant properties in biological systems,
ˈIdealˈ and ˈCarminˈ, have been patented and preventing common degenerative processes
the aim of this study was to research their (Padayatty et al., 2003). Of course, the values
quantity and quality potential. Among the are influenced by variety, crop technology and
characteristics of the bell pepper fruits, the harvest time. That is why, in this study, for
length and the diameter stand out and these biochemical analysis fruits were harvested in
characteristics have been considered extremely two different times: green stage and red stage.
important for commercialization (Blat et al., Their attractive red colour is due to the profuse
451
synthesis of various carotenoid pigments during Physico-chemical analysis
ripening. These include oxygenated carotenoids Researches have been carried out in the
such as capsanthin, capsorubin and crypto- Research Center for Studies of Food Quality
capsin, which are exclusive to this genus and and Agricultural Products from University of
have been shown to be effective free radical Agronomic Sciences and Veterinary Medicine
scavengers (Matsufuji, Nakamuro, Chino, of Bucharest, Romania. Physico-chemical
Takeda, 1998). Also results of Kim et al. analyses were performed on unripe fruit (green
(2016) shows that intake of sweet pepper might stage) at the beginning of August and on ripe
be helpful for lowering the risk of diseases fruits (red) in September.
caused by oxidative stress. The aim of this For each variety were harvested an increase
study was to evaluate the yield and quality number of sampling so that the confidence
potential of those two new Capsicum varieties, interval of the sample to get wider. Each
ˈIdealˈ and ˈCarminˈ. analysis was an average sample of several
fruits.
MATERIALS AND METHODS Ascorbic acid content was determined with
HPLC - Aqilent Technologies 1200 Series
To assess the morphological characteristics, equipment. Ascorbic acid was determined by
agronomic performance and biochemical using methods described by Jones and Hughes
compounds, varieties were grown under (1983), Deepa et al. (2006), Topuz et al.
protected environment within V.R.D.S. Buzau (2007), Ghasemnezhad et al. (2011) with slight
site during spring-autumn of 2017. During modification. From average sample of fresh
summer, from June to end of August, the fruits were taken 1 g and mixed with a pinch of
greenhouse was covered with shading net with quartz sand.
a shading degree of 40%. The mixed sample was homogenized with 10
The seeds were sown on 07.03.2017 and mL of metaphosphoric acid. The samples were
transplanted in the greenhouse at the beginning kept in the dark for 45 minutes at 4°C. The
of May, on 03.05.2017. Planting was made in extract has been centrifuged for 1 minute at
strips in the following formula 70 cm x 35 cm 1000 rotation/minute.
and 1.2 m between the strips. The supernatant was filtered through a filter
The experiment was organized in randomized Agilent RC 0.2 µm and then it was injected in
blocks, in two rehearsals and the observations HPLC. The samples were analysed in duplicate
were made on 100 plants for each variety. Both for each varieties and were expressed in
varieties have received standard cultural mg/100 g.
practices, a special care was made to handle the Total soluble solids (TSS) and titratable
bell pepper shoots and to remove of the first acidity (TA). TSS is an index of soluble sugar
floral button. content in fruit. Soluble solids were determined
During vegetation period, there have been from bell pepper juice using refractometer
recorded some agronomic traits like: earliness, Kruss (% Brix).
number of fruits/plant, weight of fruits, weight Titratable acidity (TA) was determined using
of receptacle, weight of fruit pulp, the thickness 10 g of ground bell pepper diluted in 50 ml of
of pulp. distilled water. The titratable acidity of peppers
To highlight the variability of the two varieties, was determined by titration with 0.1N NaOH to
a morphological characterization was pH 8.1 (Sadler and Murphy, 2010; Serrano et.
performed using the DUS test, according to al, 2010).
European Union - Community Plant Variety For titration was used automatic titrator
Office (2007) (CPVO-TP/076/2). TitroLine easy. The results were expressed in
The characteristics of fruits analyzed were as milligrams citric acid/100g.
follows: unripe colour fruit, cross-section fruit, The dry matter was determined by drying stove
number of locules, fruit shape at blossom end, for 15 hours at 105°C, until the samples reach
fruit shape at pedicel attachment and thickness constant weight.
of the pulp.
452
RESULTS AND DISCUSSIONS The average weight of the fruits is 135.2 g and
the thickness of the flesh is on average 6.8 mm.
Vegetable Research-Development Station The average weight of the receptacle has been
Buzău had undertaken research under the 19.3 g and the average pulp weight 115.9 g.
breeding program which aimed primarily in The low temperature during summer nights
obtaining new varieties of bell pepper suitable causes the fruits to form a pointed end at the
for greenhouse growing, adapted to pedo- apex of the fruit.
climate conditions of Romania and meet the ˈCarminˈ variety (Figure 2) has a compact
requirements of farmers and consumers. bush, with a height over 90 cm, very well
The new varieties obtained, ˈIdealˈ and branched. The fruit set percentage is medium in
ˈCarminˈ, were morphologically analyzed the hot summer days and high during autumn,
according to DUS test and the result obtained September-October period.
are shown in the Tabel 1.
Tabel 1. Main fruit characteristics

Variety ˈIdealˈ ˈCarminˈ
Fruit: colour before maturity 3 (green) 3 (green)
Fruit: colour at maturity 8 (red) 8 (red)
Fruit: shape in cross section 3 (circular) 3 (circular)
(at level of placenta)
Fruit: number of locules 2 (predominantly 4 (equally
two) three and four)
Fruit: depth of stalk cavity 5 (medium) 7 (deep)
Fruit: shape of apex 2 (moderately 4 (moderately
acute) depressed)
Fruit: thickness of flesh (mm) 7 ( thick) 6 (medium)
ˈIdealˈ variety had been characterized by

earliness, 12 days earlier than ˈCarminˈ variety Figure 2. Crop view ˈCarminˈ variety
and with a high average yield of 3.5 kg/plant
compared to ˈCarminˈ variety 2.59 kg/plant There has been, in average, a number of 16
average yield. Although ˈCarminˈ has less fruit, fruits/plant. Fruits have a pleasant commercial
the fruit average weight is higher. appearance, slightly waxed, with three of four
ˈIdealˈ variety (Figure 1) has a vigorous plant lobes. The fruits have light green colour when
growth habit, with a height over 1 m, strongly unripe and red when they reach physiological
branched, with a high percentage of fruit set maturity. The average of fruit weight has a
even at high temperatures during hot days in value of 152.3 g, and the average of pulp
the summer. thickness has a value of 5.78 mm. The average
of receptacle has weighted 22.4 g and the pulp
had 129.9 g.
In Figure 3 can be seen fruits of ˈIdealˈ and
ˈCarminˈ variety in different colours during
ripening period.
Figure 1. Crop view ˈIdealˈ variety

Figure 3. Different colour of fruits during ripening
The average number of fruits per plant has been
26 fruits. Fruits are slightly conical with two In this study, the fruit quality of the sweet
locules, light green before maturity and red peppers variety was assessed by: ascorbic acid
when ripe.
453
content, dry matter, total soluble solids and Physico-chemical analyses of total soluble
titratable acidity. The quality indicators were solids (TSS), titratable acidity (TA) and dry
different in both ripening stages: green and red. matter values are presented in Table 2.
The amount of ascorbic acid was higher in Table 2. Physico-chemical analyses
ˈCarminˈ variety, in both stages green and red, Stage Analyse ˈIdealˈ ˈCarminˈ
as seen in Figure 4, values vary from 164.31 Green TA (citric acid) %
TSS °Brix
0.23 mg/100g
4.87
0.17 mg/100 g
4.52
mg/100 g when green stage and 189.27 mg/100 Dry matter (g) 7.99 6.23
g when ripe. Red TA (citric acid) % 0.28 mg/100g 0.29 mg/100 g
TSS °Brix 7.25 6.75
ˈIdealˈ variety records a value of 79.29 mg/100 Dry matter (g) 9.43 9.01
g when fruits are unripe and green and an
increased value of 122.5 mg/ 100 g when fruits As seen in Table 2, the quantity of titratable
have reached physiological maturity. acidity increased with ripening of varieties.
While the fruit ripens, the metabolic reactions
increase, increasing the concentration of
organic acids involved in the Krebs cycle.
Apart from this, these acids make up the
energetic reserves and the metabolic reactions
that involve the synthesis of pigments, enzymes
and other materials and the degradation of
pectins and celluloses, which are essential for
the ripening process. The organic acids are
active substances during ripening in these
Figure 4. Variation of ascorbic acid during alterations (Chitarra and Chitarra, 1990). For
ripening period the totally green bell peppers, these organic
acids are present in small quantities, as the
As fruit ripens, an increase in ascorbic acid ripening process has not yet started, presenting
content was observed, these results have been differences in relation to the other stages of
in assent with previous studies (Howard et al, ripening. The same behaviour was observed by
1994; Deepa et al., 2006; Osuna et al., 1998; Molinari et al. (1999) and by Ghasemnezhad et
Marin et al., 2004). Smirnoff and Wheeler al. (2011).
(2000) states that the role of ascorbate content Total soluble solids it is also increasing during
as a photoprotector has been reported for leaves ripening period on both sweet pepper varieties.
acclimated to high light, which have a higher As fruits starts ripening there is an increased of
ascorbate concentration than leaves grown at total soluble substance due to the degradation
low intensity. The amount and intensity of light or biosynthesis of the polysaccharides and the
during the growing season have also been accumulation of sugars. The metabolic
described to have a definite influence processes related to the advance of ripening are
increasing the ascorbic acid formed. The light directly influence by the level of TSS, where
exposure could explain the increase in ascorbic fruits in advance stages of ripening present the
acid content of red fruits as compared to the highest levels of TSS. These increases in
green ones (Lee, 2000). soluble sugars and acidity have also been found
To provide antioxidant protection, a during physiological ripening in many cultivars
Recommended Dietary Allowance (RDA) of of both hot and sweet pepper, being correlated
vitamin C is 90 mg/day for adult men and 75 to colour changes (Lyon et al., 1992;
mg/day (Bendich A., 2001), individuals who Ghasemnezhad et al., 2011; Behera et al., 2004;
smoke require 35 mg/day more. According to Orowski et al., 2004; Tadesse et al., 2002,
the results obtained for ˈCarminˈ and ˈIdealˈ Serrano et al., 2010).
variety, it was learned that it can provide the The amount of dry matter is an important
recommended dietary allowance and even more quality attribute of bell pepper. High values of
with just 100 g/day. this parameter induce a better flavour and taste.
It has been registered an increase in green stage
on ˈIdealˈ variety from 4.87 g to 7.25 g in the
454
red stage and referring to ˈCarminˈ variety, the Medicine Washington, DC: National Academy Press,
registered differences oscillated between 4.52 2000 ISBN: 0-309-06935-1. Nutrition, 17 (4): 364.
Blat S.F., Braz L.T., Arruda A.D.S., 2007. Avaliação de
g to 6.75 g. Concluding, ˈIdealˈ variety has a híbridos duplos de pimentão. Horticultura Brasileira,
stronger flavour and a more pleasant taste than 25: 350-354.
ˈCarminˈ variety. Fruits who have reached Burzo I., 2015. Compoziția plantelor medicinale și
physiological maturity have a higher amount of alimentare din flora spontană și cultivată. Editura
dry matter, which is probably due to high Elisavaros, București. 166-167.
Chitarra M.I.F., Chitarra A.A., 1990. Po´ s-colheita de
temperature during summer, which allows an frutas e hortaliças. Fisiologia e manuseio, 293.
increase nutrition and plant metabolism. Deepa N., Kaur C., Singh B., Kapoor H.C., 2006.
Similar results were recorded by Amalfitano et Antioxidant activity in some red sweet pepper
al. (2017), in Naples, Italy. cultivars. Journal of Food Composition and Analysis,
19 (6-7): 572-578.
European Union - Community Plant Variety Office
CONCLUSIONS (2007) Protocol for distinctness, uniformity and
stability tests of Capsicum annuum L. (sweet pepper,
The two new varieties obtained at Vegetable hot pepper, paprika, chili). Available at
Research-Development Station Buzău, ˈIdealˈ http://www.cpvo.europa.eu/documents/TP/vegetales/
and ˈCarminˈ, have shown genetic stability in TP_076-2_CAPSICUM_ANNUUM.pdf.
Ghasemnezhad M., Sherafati M., Payvast G.A., 2011.
terms of phenotypic expressiveness of the main Variation in phenolic compounds, ascorbic acid and
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456
INFLUENCE OF PLANT MANAGEMENT SYSTEMS ON GROWTH AND

FRUCTIFICATION OF TOMATO PLANTS IN PROTECTED CULTURE
Gheorghița HOZA1, Maria DINU2, Rodica SOARE2, Alexandra Dida BECHERESCU3,
Ionuț Alexandru APAHIDEAN4, Dorel HOZA1
1
59 Marasti Blvd, Bucharest, Romania
2
University of Craiova, 13 Alexandru Ioan Cuza Street, Craiova, Romania;
3
from Timisoara, 119 Calea Aradului Street, Timisoara, Romania
4
University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca,
3-5 Manastur Street, Cluj-Napoca, Romania
Abstract
Tomatoes have a high capacity to form shoots on the stem which is why plants can be managed with multiple stems,
thus influencing the increase in production and decrease in the number of plants per hectare. Research was conducted
in the teaching field of the Faculty of Horticulture in Bucharest, within a prolonged cycle in solarium, during 2016-
2017. Two varieties were used, Belmonte and Canestrino, and two hybrids, Cinto F1 and Clarabella F1, managed with
one and a double stem. Stem management was performed through eliminating the portion of the plant from above the
cotyledonal leaves, during the seedling phase, after the formation of the first pair of leaves, the percentage of stem
formation being over 90 %. The planting was made at a distance of 0.8 m/0.4 m for the plants with one stem, resulting
in 3.1 plants/m2, and at 1 m/0.4 m for the plants with a double stem, resulting in 2.5 plants/m2. The reduction in the
number of plants was of 20% for the ones with double stem. The largest production of fruit per plant was obtained for
the tomatoes with a double stem: 7.2 kg for the Belmonte variety and 7.2 kg for Canestrino, 6.9 kg for the hybrid
Clarabella F1 and 6.4 kg for Cinto F1. The plants with one stem recorded a production of 2.6-2.9 kg/plant. The
production per square meter was directly influenced by the production obtained per plant; thus, the plants with double
stem recorded values of 16-18 kg, while the tomatoes with one stem produced 8.06-8.70 kg.
Key words: double stems, pinching off, production, seedling.
INTRODUCTION currently on improving the culture technology.

From a biological point of view, tomatoes are
Tomatoes (Lycopersicon esculentum Mill) have herbaceous annual plants, with a very good
their origins in South America and, compared capacity to form shoots, which has led to
to other vegetable species, have the largest numerous researches regarding the increase of
cultivation area in the world, being very the production per plant and per unit of area,
appreciated by consumers. The fruits are rich in without affecting the production quality. In a
vitamins, minerals, amino acids and pigments culture tomatoes are usually managed with one
(Dinu et al., 2017; Soare et al., 2015) and poor stem; however numerous researches have been
in calories, being considered very healthy for performed in which plants have been managed
the human body. The biochemical composition with 2, 3 and 4 stems, using only cultivars with
of the tomato fruit is influenced, amongst other undetermined growth. Obtaining several stems
factors, by the position of the fruit within the can be achieved through pinching off the plant
inflorescence. The fruit in the middle has the above the cotyledonal leaves (Hoza and
highest content of polyphenols. The sugar Stanciu, 2012; Mouaro et al., 2014), pinching
content increases simultaneously for the fruits off above the first pair of real leaves (Mouaro
ranging from the base to the top of the et al., 2014), pinching off above the second
inflorescence while fruit size decreases inflorescence (Franco et al., 2009) or through
(Coyago-Cruz, 2017). Researches performed removing the growth peak of the seedling at
on tomatoes are diverse and attention is planting and using the first 2 shoots at the base
457
of the plant (Hoza et al., 2011; Hoza, 2013). MATERIALS AND METHODS
The research conducted by Mouaro et al.
(2014) proved that plants managed with 2 The experiment was conducted in the teaching
stems generate larger productions (26.5 kg m-2) field of the Faculty of Horticulture Bucharest,
compared to plants managed with 3 and 4 in solarium, during 2016-2017. The main
stems, for which the production was smaller purpose of the experiment was to determine the
(19.5 kg m-2). Comparing the production for influence of the number of stems and of the
the double stem of the same plant, the cultivar on plant growth and fructification, in
differences were not significant. According to the climatic conditions of the reference area,
Rahmatian et al., 2014, grafted tomatoes and the introduction of this particular plant
managed with 2 stems in a hydroponic system management system into the tomato culture
formed more fruits per plant (46.04) than the management. Two varieties were used,
ones with only one stem (27.43); the Canestrino and Belmonte, highly perishable,
production per plant was larger (4.45 kg) with superior characteristics regarding the taste
compared to plants with one stem (3 kg), the and large fruits and two F1 hybrids, Clarabella
fruits were firmer and of a higher quality, but and Cinto, very productive, with large,
the average fruit weight was lower by 12 %. In uniform, firm fruits with high resistance to
addition, grafted plants with a double stem various pathogens. Plants were managed with
accumulated a larger quantity of dry substance one and a double stem and 6 inflorescences
in various plant organs. Tomatoes with 2 and 3 were kept on each stem. Double stems were
stems formed from shoots led to an increase in obtained through eliminating the portion of the
the number of fruits (Hoza and Stanciu, 2012) plant from above the cotyledonal leaves when
and in the commercial production compared to the tomato seedlings formed the first pair of
plants with one stem. However their fruits were real leaves. The buds at the base of the
slightly smaller and the plants did not cotyledons formed 2 shoots that later became
registered significant differences regarding double stems. Specific maintenance works
quality and number of stems; plants conducted were applied for the tomato seedlings up to the
with 3 stems also extend the harvesting period planting moment.
(Mbonihankuye et al., 2013). For cherry Field preparation was made through soil
tomatoes, plant management with 2 stems is a loosening and crumbling works and
current practice in southern Spain, stem fertilization with Orgevit 2 t/ha incorporated
planning being made after the second into the soil before planting.
inflorescence forms. According to Franco et al., The culture was established with seedling pro-
2009, for cherry tomatoes cultivated in duced in warm greenhouse, of 55 days of age.
greenhouse, managed with 2 stems on which During seedling production period, plants
are kept 2 side-shoots with 2 inflorescences on managed with one stem were kept in colder
each, the production and number of fruits spaces (16-18°C) compared to the ones with a
increase per unit of area, without significantly double stem, in order to reduce the growth
affecting the average fruit weight. By analyzing difference and to ensure the planting could be
the correlation between the number of stems made at the same time. This aspect is aimed be
and the tomato production, it has been observed a subject to further research themes. Planting
that plants managed with 2 stems have had a was made during the first decade of April for
higher production and number of fruits, but the both experimental years, at a distance of 0.8
average fruit weight and the commercial m/0.4 m for plants with one stem, resulting in
production have been slightly lower for the 3.1 pl./mp, and at a distance of 1 m/0.4 m for
plants with 2 stems compared to the ones with plants with a double stem, resulting in 2.5 pl./mp
on stem (Ece and Darakci, 2007). Plant and a reduction in the number of plants of 20%.
management with more stems in association Plants with a double stem were led as a V-shape
with different doses of nitrogen demonstrated towards the spaces between rows in order to
that the largest production has been registered allow light to penetrate through more efficiently.
at plants with 2 stems and 256 kg N ha-1 During the vegetation period, all lateral shoots
(Hossain, 2007). were eliminated, plants were supported on
458
strings and optimal growth conditions were experiment; average fruit weight was
ensured: temperature, humidity and light determined through harvesting fruit per variant,
according to the phenophase requirements. counting and weighting the fruits at each
Phased foliar fertilizations were applied to harvest and calculating the average weight.
ensure the nutrition necessary for the optimal Measurements were made also regarding plant
growth and fructification of the tomato plants growth, determined by the height at the end of
using: Codicevo at planting and Energevo at the vegetation period, distance from the soil for
fruit formation. During growth and fruit the first inflorescence and average distance
maturation Finalevo and Rezistevo were between inflorescences. During the fructifica-
applied. Fertilizer composition was as follows: tion period, the number of fruits per plant,
- Codicevo 19:19:19+ME 0.2%, with a content number of fruits per square meter, average fruit
of NPK of 19:19:19, S 1.4%, MgO 1.8%, B weight and the production per plant and per
0.1%, Fe 0.07%, Mn 0.03%, Zn 0.03%, Cu square meter were measured, according to the
0.006% and Mo 0.002%, that favors a rapid type of cultivar and plant management system
and improved absorption of the soil nutrients; used. The primary data was processed
- Energevo 9:53:9 0.2 %, with a content of N statistically by the variant analysis method.
9%, P2O5 53%, K2O 9%, MgO 2.5%, Fe 0.1%,
S 1.0%, B 0.1%, Mn 0.05%, Zn 0.1%, Cu RESULTS AND DISCUSSIONS
0.05% and Mo 0.005%, that increases plant
resistance to various stress factors and Regarding the vegetative growth of the tomato
accelerates the ongoing of vegetation phases; plants it was observed that between the plants
- Finalevo 5:14:42 0.3 %, that contains N 5%, with one stem and the ones with a double stem
P2O5 14%, K2O 42%, MgO 2.5%, S 6%, B small differences were recorded, but the
0.1%, Fe 0.1%, Mn 0.05%, Zn 0.1%, Cu 0.05% cultivar had a stronger influence on some
and Mo 0.005%, and improves the quantity and indicators (Table 1). Regarding the distance
quality of the harvest; from the soil for the first inflorescence, it had
- Rezistevo 0.3 %, that has N 14%, CaO 18%, higher values for the varieties Camestrino and
K2O 1.7%, MgO 4.2%, Mn 0.05%, B 0.2%, Zn Belmonte, compared to the F1 hybrids
0.01% and Mo 0.005%, applied at an interval Clarabella and Cinto. The average distance
of 15 days to diminish the effects of calcium between inflorescences was slightly larger for
deficit and increase the fruit resistance to the plants with two stems, except for the
cracking, transport and storage. variety Canestrino, for which the average
The experiment was bifactorial, organized in distance between inflorescences was smaller
random blocks, with three repetitions. The for the plants with double stem. Average plant
following elements were used: four cultivars height was smaller for the varieties Canestrino
with large fruits and undetermined growth and and Belmonte with a double stem, while
two plant management systems - S1 with one Clarabella and Cinto had lower values for the
stem and S2 with a double stem. plants with one stem; on average, plants with
Measurements were made on five plants from double stems slightly surpassed the average
each repetition, selected at the beginning of the height of the ones with just one.
Table 1. Synthesis of the results regarding the growth of tomato plants with one and a double stem
Cultivar Plant management Distance from soil for Average distance between Plant height
system first inflorescence (cm) inflorescences (cm) (cm)
Canestrino S1 36.2 36.9 221.4
S2 35.7 35.3 214.1
Belmonte S1 32.6 26.5 175.0
S2 34.2 28.9 173.6
Clarabella F1 S1 24.4 31.5 187.2
S2 24.9 33.5 192.5
Cinto F1 S1 23.6 31.2 185.8
S2 22.2 32.9 196.1
Average (mt) S1 29.2 31.5 192.4
S2 29.3 32.7 194.1
459
The interaction between the cultivar and meter, the average increase of fruit varying
number of stems was obvious in the potential between 32% for the hybrids and 77% for he
of the fructification capacity, the plants with varieties, having an average value of 45%. The
double stems producing more fruits than the number of fruits per square meter was between
ones with only one stem (Table 2). 100 fruits for Clarabella F1 and 113.8 for
The number of fruit per plant recorded an Belmonte, values slightly lower than the ones
average increase amongst the studied varieties obtained by Mouaro et al., 2014, respectively
of 80%; for the variety Canestrino the increase 121.5 and 125 fruits per m-2, dependant on the
was more than double compared to the other stem planning.
varieties, respectively 119 %, while for the two The cultivar’s influence on the fruit production
hybrids it was of 64%. was poorly highlighted, both regarding the
The number of fruits per plant with a double number of fruits per plant and the number of
stem was between 40 and 45.5, being close to fruits per square meter.
the data recorded in the professional literature The average fruit weight (Table 2) was lower
(Ece and Darakci, 2007; Hoza and Stanciu, for all cultivars with a double stem compared to
2012; Rahmatian et al., 2014.). the ones with one stem, fact proved by the
The number of fruits obtained per square meter professional literature (Ece and Darakci, 2007;
depended on the number of fruits per plant and Hoza and Stanciu, 2012; Rahmatian et al.,
the number of cultivated plants per square 2014).
Table 2. Number of fruits per plant and per m2 depending on the cultivar and number of stems
Difference
S1(one stem) S2 (double stem) between
S1 and S2, %
Cultivar
Average No of No of
Average fruit No of No of
fruit fruits fruits plant-1 m-2
weight, g fruit/pl-1 fruits/m-2
weight, g plant-1 m-2
Canestrino 133.7 *** 20.2 o 62.6ooo 132.6*** 44.3 * 110.8 N +119 +77
ooo ooo
Belmonte 111.2 25.0 N 77.5 ** 109.9 45.5*** 113.8 ** +82 +47
Clarabella F1 117.8oo 24.4 N 75.6 N 116.6ooo 40.0ooo 100.0ooo +64 +32
Cinto F1 126.3 ** 27.0 N 83.7 *** 124.7*** 44.2 * 110.5 N +64 +32
Media (mt) 122.5 24.2 74.9 121.0 43.5 108.8 +80 +45
DL 5% 2.20 3,41 1,83 1.13 0.73 2.45 - -
DL 1% 3,33 5.16 2,77 1.72 1.11 3.72 - -
DL 0.1% 5.31 8.23 4,41 2.74 1.77 5.92 - -
The productive capacity of the tomato plants was noted that plants with one stem formed 3
was influenced by the cultivar, but especially kg plant-1, while the ones with double stems
by the number of stems (Table 3). For the formed 5.3 kg plant-1. Franco et al., 2009,
plants managed with a double stem, the showed that for cherry tomatoes cultivated in
production was superior compared to one stem greenhouses, with a double stem and with two
plants as shown also by Hossain (2007) and shoots on each stem, the production per unit of
Mouaro et al. (2014). For this experiment, the area increased.
largest production per plant was recorded for The present experiment confirms this fact, the
the variety Canestrino with a double stem, 5.9 production for the plants with two stems being
kg plant-1, the production increase being of larger than for the tomatoes with one stem,
+3.2 kg plant-1 compared to the one stem with values varying from 11.6 kg m-2 for Cinto
plants. Conversely, the smallest was obtained F1 and 14.7 kg m-2 for Canestrino. The average
from the hybrid Clarabella, 4.6 kg plant-1, the production of the cultivars with one stem was
production increase being of +1.7 kg plant-1 for of 9.1 kg m-2, while for the ones with double
two stem plants. Analyzing the average stems was of 13.2 kg m-2.
production per plant for the used cultivars, it
460
Table 3. Tomato production per plant and m2 depending on the cultivar and number of stems
S1 (one stem) S2 (double stem) Difference between S1 and S2
Cultivar Production Production
kg plant-1 kg m-2 kg plant-1 kg m-2 kg plant-1 kg m-2
Canestrino 2.7 N 8.4oo 5.9 ** 14.7 *** +3.2 +6.3
Belmonte 2.8 N 8.6o 5.0 N 12.5oo +2.2 +3.9
Clarabella F1 2.9 N 8.9 N 4.6oo 11.6ooo +1.7 +2.7
Cinto F1 3.4 * 10.6 *** 5.5 N 13.8 ** +2.1 +3.2
Average (mt) 3.0 9.1 5.3 13.2 +2.3 +4.1
DL 5% 0.34 0.38 0.34 0.39 - -
DL 1% 0.51 0.57 0.51 0.59 - -
DL 0.1% 0.82 0.92 0.82 0.95 - -
CONCLUSIONS International Multidisciplinary Scientific Geo

Conference SGEM Conference Proceedings, ISSN
1314-2704, June 29-July 5, Vol. 17 (Ecology,
The research conducted with four tomato Economics, Education and Legislation), p. 93-100.
cultivars and two plant management systems, Dinu M., Soare R., Dumitru M.G., 2017. The effect of
one with one stem and one with a double stem, foliar fertilization with organic products on some
showed that tomato plants react very well when nutritional value during post-harvest storage of
they have multiple stems. tomatoes (Lycopersicon esculentum Mill). Archivos
Latinoamericanos de Nutrición.Número 3, Volumen
From a vegetative point of view, no significant 67.
differences were recorded between plants with Ece Ali, Nilay Darakci, 2007. Determination of
one stem and plants with a double stem. Relationships between Number of stem and yield of
However, on average, plants with a double tomato (Lycopersicon lycopersicum L.) Asian Journal
stem had a slightly higher growth (194.1 cm) of Plant Sciences, 6 (5): 802-808.
Hoza Gh., 2013. Researche regarding the influence of
compared to plants with just one (192.4). various pruning systems at Chornichon cucumber
The fructification process was influenced by cultivated in solarium, Journ. Hort. For. Biotec.
the plant management system, plants with USAMVB Timisoara, vol.17 (1), 219-222.
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plant and by 45% more fruits per square meter regarding management systems for plants with two
stems, cultivated in solar, Lucrări ştiinţifice, Seria
than the single stem plants. Horticultură, vol 54 (2), 57-160.
The largest average production per plant (5.3 Hoza Gh., Stanciu L.G., 2012. Research regarding the
kg) and the largest production per square meter influence of tomato plant management for cultures
(13.2 kg) were obtained also from the double grown in solarium, in extended production cycle.
stem plants, result justified by the presence of a Analele Universităţii din Craiova, vol.XVII (LIII),
211-216.
double locus of fructification on the plant. José L.F., Díaz M., Diánez F., Camacho F., 2009.
Average fruit weight was slightly smaller for Influence of different types of pruning on cherry
the double stem plants (121.0 g) compared to tomato fruit production and quality Journal of Food,
the plants with one stem (122.5 g), however Agriculture & Environment Vol. 7 (3 and 4): 248-253.
this result cannot be used as an argument that Mbonihankuye C., Kusolwa P., Msogoya T.J., 2013.
Assessment of the Effect of Pruning Systems on Plant
plants with one stem ensure an obvious Developmental Cycle - Yield and Quality of Selected
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Dinu M., Hoza Gh., Becherescu A., 2017. Antioxidant Aksoy U. (Eds.) Proceedings of the 4th ISOFAR
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Rahmatian A., Delshad M. Salehi R., 2014. Effect of Soare R., Dinu M., Roşculete E., 2015. The influence of
Grafting on Growth, Yield and Fruit Quality of Single the hybrid on early tomatoes production. Annals of
and Double Stemmed Tomato Plants Grown the University of Craiova, Agriculture, Montanology,
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462
EFFECT OF GRAFTING ON YIELD AND FRUIT QUALITY OF PEPPER

(CAPSICUM ANNUUM L.) GROWN UNDER OPEN FIELD CONDITIONS
Volkan ERGUN, Hakan AKTAS
Suleyman Demirel University, Faculty of Agriculture, 32260, Isparta, Turkey
Abstract
The aim of the study, determining of grafting could improve the agronomic behaviour of pepper (Capsicum annuum L.),
an open field experiment was carried out to determine growth, yield and fruit quality of long type pepper hybrid
cultivar (ʻEfilʼ). As a scion plant material was used ʻEfil F1ʼ and rootstock ʻGuclu F1ʼ. ʻEfil/Efilʼ (ʻScion/Scionʼ),
ʻEfil/Gucluʼ (ʻScion/Rootstockʼ) and non-grafted ʻEfilʼ (ʻSiconʼ) were used as the grafted combination. According to
experiment, grafted plants were taller than control (non-grafted). Total yield, fruit number, fruit flesh firmness, fruit
weight and stem diameter were influenced by rootstock and grafting. Grafted pepper produced 16% more yield than
control plants for 'Efil/Guclu'. A similar trend was also observed on 'Efil/Efil'. The lowest yield recorded on ʻEfilʼ (non-
grafted).
Key words: grafting vegetable, Capsicum annuum, scion, rootstock.
INTRODUCTION tremendously (Lee et al., 2010). At present,

grafting is mainly used in order to reduce
Pepper (Capsicum annuum L.) is a crop of high infections by soil-borne pathogens and to
economic importance in Turkey. The area of enhance the tolerance against abiotic stresses
pepper grown in Turkey is 1.141.216 da with (King et al., 2008; Louws et al., 2010; Rivero et
3.414.852 million tons of pepper production al., 2003).
(TUIK, 2016). Pepper growing is getting more One way of avoiding or reducing losses in
difficult day by day, because of crop damage production caused by pathogens in high-
due to the specific pathogens, Phytophthora yielding genotypes would be to graft them onto
capsici, Verticillium dahliae and Meloidogyne rootstocks capable of reducing soil-borne
spp. in soil (Morra and Bilotto, 2006; Myung et diseases and increasing yield and fruit quality
al., 2006). (Lee, 1994). In order to prevent soil-borne
One of the major goals in pepper breeding is diseases in continuous cropping, peppers are
the development of a cultivar completely generally grafted onto the rootstocks that are of
resistant to soil-borne diseases. However, that the same species as scions (C. annuum L.) that
is very difficult to achieve and requires much have resistance to Phytophthora blight (King et
time and effort. al., 2010). It was reported that grafting of
For the alleviation of soil-borne diseases, peppers also improved tolerance to high salt
cultural practices such as crop rotation and conditions (Chung and Choi, 2002) and low
sanitation are recommended, but pesticide is temperature (Jang et al., 2008).
generally applied to control the diseases (Kim The aim of this study was to investigate and
et al., 2010; Semi et al., 2010; Tran and Kim, evaluate the agronomic performance of hybrid
2010; Yeon et al., 2008). Grafting is an pepper (scion) under open field conditions
environment-friendly alternative method for following grafting on pepper rootstocks, in
disease control (Oka et al., 2004; Rivard and comparison with un-grafted and scion self-
Louws, 2008). Grafting scions onto resistant grafted (scion/scion) plants.
rootstocks makes it possible to control soil-
borne diseases and increase yield of the MATERIALS AND METHODS
susceptible cultivar (Lee and Oda, 2003).
Recently, the cultivated area of grafted The F1 hybrid, long type ‘Efil’ (Asgen,
Solanaceae and Cucurbitaceae have increased Turkey) was grafted on commercial rootstocks,
463
‘Guclu’ (Graines Voltz, Türkiye). Un-grafted All data were statistically analyzed by ANOVA
‘Efil’cultivar and itself grafting ‘Efil/Efil’ were using the MINITAB software package.
also used as control.
The cleft grafting was realized when rootstocks RESULTS AND DISCUSSIONS
and grafts showed six and two true leaves,
respectively. Grafted and un-grafted pepper Plant height was not significantly influenced by
plants were transplanted on 10 April 2016 in grafting combination while plant diameter
open field condition on the Experimental Farm affected significantly by grafting (Table 1).
of Suleyman Demirel University. Grafted plants were 9 and 15% thinner scion
The experimental soil was loamy (Bouyoucos, diameter than control for ‘Efil/Efil’ and
1962) having pH 7.9 (1:2.5 soil to water ratio), ‘Efil/Guclu’, respectively.
9.5 % CaCO3, 1.1% organic matter (Jackson, Among the combination, the greatest plant
1962), 15.9 mg kg-1 NaHCO3 extractable P diameter was recorded on non-grafted plant
(Olsen et al., 1954), 125, 266, 375 mg kg-1 1 N ‘Efil’. The plant height was slightly influenced
NH4OAC exchangeable K and Ca and Mg by grafted (Table 1).
(Knudsen et al., 1982). DTPA extractable Fe,
Cu, Zn and Mn concentrations (Lindsay and Table 1. Effect of grafting combination
on plant diameter (mm) and plant height (cm)
Norwell, 1978) were 2.9, 0.55, 0.89 and 11.9
mg kg-1, respectively. Plant diameter Plant height
Treatments were arranged in a randomized (mm) (cm)
12.83 A ± 0.14 58.55 ± 0.82
complete-block design with three replications, Efil
Efil/Efil 11.65 B ± 0.15 60.82 ± 0.77
each consisting of 30 plants. Plants were grown
Efil/Güçlü 10.87 C ± 0.20 59.23 ± 0.97
in single rows (1.0 m × 0.50 m) at a plant
density of 2.5 plants m-2.
Irrigation was applied by drip-irrigation and There was no differences determinate plant
was scheduled using tension meters to ensure fresh and dry weight between grafted and non -
that water was non-limiting (the high and low grafted plants (Table 2).
tension set points were –30 kPa and –1 kPa,
Table 2. Effect of grafting combination
respectively).
on plant fresh and dry weight (g)
The irrigation frequency was arranged every
morning. The Following nutrient content; Plant fresh
weight Plant dry weight
(mg/L): N: 100; K: 50; P: 60; Mg: 30; Ca: 30;
(g) (g)
Fe: 3.0, Mn: 2.0; Zn: 0.25; B: 0.70; Mo: 0.05
Efil 190.7 ± 6.86 40.59 ± 1.39
was used during the experiment.
Efil/Efil 190.6 ± 5.75 40.58 ± 1.70
During the experiment, pH 6-7 and EC 1.2-1.8
179.2 ± 6.66 36.23 ± 1.22
were set. Efil/Güçlü
Plants were kept free from weeds, insects, and
diseases using standard growing management. Generally, it has been reported that grafting
The experiment was terminated after 120 days promotes vegetative growth at different levels
from transplanting, plant height was recorded dependent on rootstock (Colla et al., 2008).
on eight plants per plot. Mature green fruits Promoted vegetative growth (plant height) was
were harvested regularly every week. explained by the vigorous root system of
Fresh weight of marketable fruits, fruit number, rootstocks, which are often capable of
length, and diameter were recorded on 20 absorbing water and plant nutrients more
plants per plot. Fruits that were deformed or efficiently than scion roots and serve as a good
badly misshapen were considered supplier of endogenous hormones (Kato and
unmarketable. Fruit shape index (SI), defined Lou, 1989).
as the ratio between width and length was also But, in this study, we could not determinate
calculated. Three representative marketable highly differenceson vegetative grown such as
fruits of each plot were analyzed for fruit plant height and fresh weight etc., and between
quality parameters. grafted and non-grafted plants.
464
This situation can be explained by the fact that (Zijlstra et al., 1994), and enhancement of scion
there is no good combination between the scion vigor (Leoni et al., 1990).
and the rootstock. The joint action of some or all of these
processes could explain the higher yield in
Table 3. Effect of grafting combination on fruit length pepper from grafted plants. There are some
(cm) and fruit stem length (cm) reports that certain rootstocks may cause
Fruit length Fruit stem length deterioration in fruit quality (Lee, 1994).
(cm) (cm)
Efil 19.12 A ± 0.19 2.117 ± 0.05 CONCLUSIONS
Efil/Efil 18.38 B ± 0.17 2.183 ± 0.05
Efil/Güçlü 17.58C ± 0.24 2.075 ± 0.04 In the present study, some of qualities such as
fruit length, fruit stem length, fruit diameter,
Fruit length was significantly influenced by fruit flesh diameter were not affected by grafted
rootstock; whereas no significant difference combination. Therefore, the use of grafted
was observed on fruit stem length, fruit length pepper plants under open field conditions
(Table 3) fruit diameter and fruit flesh diameter would represent a potential strategy for an
(Table 4). increase in total yield and some of the soil
diseases without having remarkable
Table 4. Effect of grafting combination on fruit diameter deterioration in the taste of the peppers.
(mm) and dry fruit flesh diameter (g)
Fruit flesh REFERENCES
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23.475 ± 0.33 2.457 ± 0.04 making particle size analysis of soils. Agronomy
Efil
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Efil/Efil 23.168 ± 0.38 2.462 ± 0.04
Chung H., Choi Y., 2002. Enhancement of salt
Efil/Güçlü 22.339 ± 0.48 2.426 ± 0.06 tolerance of pepper plants (Capsicum annuum) by
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Total yield was influenced by rootstock, Colla G., Rouphae Y., Cardarelli M., Temperini O., Rea
E., Salerno A., Pierandrei F., 2008. Influence of
whereas fruit weight was not affected by grafting on yield and fruit quality of pepper
grafting. The highest yield was obtained by the (Capsicum annuum L.) grown under greenhouse
combination ʻEfil/Gucluʼ, while the lowest conditions. Acta Horticulturae, 782, 359-364s.
value was recorded on the control (non-grafted) Davis S.R., Liu A.R., Levi W.G., 2008. Grafting for
and ʻEfil/Efilʼ (grafted itself) (Table 5). disease resistance. Hort science 43, 1673-1676.
Jackson M.L., 1962. Soil chemical analysis, Englewood
Grafted rootstock (ʻEfil/Gucluʼ) produced Cliffs, N.J.: PrenticeǦHall, Inc.
around 12% more yield than control plant Jang Y.A., Cho Y.Y., Rhee H.C., Um Y.C., 2008.
(Table 5). Effects of rootstock and night temperature on the
growth and yield of grafted pepper (Capsicum
Table 5. Effect of grafting combination on fruit weight annuum L.). Hort. Environ. Biotechnol. 49:63-71.
(g) and total yield (kg/plant) Kato T., Lou H., 1989. Effect of rootstocks on yield,
mineral nutrition and hormonal level in xylem sap in
Fruit weight Total Yield egg plant. J. Jap. Soc. Hort. Sci. 58: 345-352.
(g) (kg/plant) Kim B.S., Kwon T.R., Hwang J.E., Lee J.M., Park
Efil 32.98 ± 0.70 0.66 ± 0.09 D.G., Ahn J.H., Kim H.Y., 2010. Resistance to
33.22 ± 0.70 0.66 ± 0.04
Phytophthora blight of commercial pepper cultivar
Efil/Efil
in Korea. Res. Plant Dis. 16: 141-147.
Efil/Güçlü 31.12 ± 1.18 0.75 ± 0.10 King S.R., Davis A.R., Zhang X., Crosby K., 2010.
Genetics, breeding, and selection of rootstocks for
It was demonstrated that grafting per se directly Solanaceae and Cucurbitaceae. Scientia Hort. 127:
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Knudsen D., Peterson G.A., Pratt P., 1982. Lithium,
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M.G., 1990. The influence of four rootstocks on some in Horticultural Plants Under Stress Conditions.
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466
RESEARCH ON THE EFFECT OF VERMICOMPOST FERTILIZATION

ON EGGPLANTS SEEDLINGS (SOLANUM MELONGENA L.)
Constantin PETRE1, Elena DOBRIN2, Elena Maria DRAGHICI2
1
Vegetable Research-Development Station Buzau, 23 Mesteacanului Street, Buzau, Romania
2
Abstract
Although worldwide, the use of vermicomposts in horticulture is a well-known technique, in Romania, this is at
pioneering level. Therefore, the present study presents the results of an experiment that aimed at determining the
influence of the application of vermicompost in the substrate in different proportions and the foliar application in
different concentrations of the vermicompost filtered macerate on the seedlings of eggplants (Solanum melongena L.).
The analysis of the results revealed that, compared to the blank variant, to which no vermicompost was applied, all the
other variants have recorded superior results in the growth and development of seedlings. The best results were
obtained in substrate fertilizer variants, 20% and 10%. Favorable results were also recorded in the variant where the
vermicompost was applied foliar as filtered macerate at a concentration of 30%.
Key words: Solanum melongena L, vermicompost, growth, eggplants.
INTRODUCTION Several studies have evaluated the impact of

the application of vermicomposts in substrate
The massive use of chemical fertilizers in mixtures for the production of seedlings, in
horticulture has already produced negative terms of plant growth and development.
effects on the quality of production, the The best responses usually occurred when the
environment and human health over the last vermicompost had a relatively small proportion
three decades, with the concept of sustainability (20-40%), higher amounts of vermicompost,
becoming a key issue for the future (Lichtfouse did not always improve plant growth (Atiyeh et
et al., 2009). Lately, amid the development of al., 2000; Hashemimajd et al., 2004; Prasanna
concepts such as integrated and biological et al., 2010).
horticulture, it has been necessary to reconsider Another possibility of using vermicomposts is
the fertilizers used in horticultural practice. An foliar application, as a filtered macerate
alternative may be the use of vermicomposts, (vermicompost tea), which can provide a rich
which, compared to conventional composts, are and active and alive bacterial flora, organic
the product of accelerated biooxidation of acids, growth regulators and mineral nutrients
organic matter by the use of high densities of that are soluble in aerial organs of plants
earthworms without the thermophilic phase (Edwards et al., 2006).
(Dominiguez et al., 1997). The present experiment aimed at testing the
Vermicompost is rich in nutrients, easily fertilizing action of the vermicompost obtained
soluble in plants (Chaoui et al., 2003). It also at VRDS Buzau over the seedlings of the
contains growth hormones, enzymes and Solanum melongena L. eggplants in order to
beneficial microorganisms and does not contain recommend it to the Romanian sapling
pathogens and toxic chemicals (Canellas et al., producers as a fertilizing support of the growth
2002). The use of vermicomposts leads to the rate and development of the plants, which
stimulation of plant immunity. Edwards et al. would lead to a better utilization of organic
(2004) established that the vermicompost has residues and waste, with implications for
reduced some fungal diseases such as Phytium reducing the production costs of seedlings and
in cucumbers, Rhizoctonia in radishes, a significant reduction of environmental
Verticillium in strawberries and Phomoposis pollution.
and Sphaerotheca fulginea in field grapes.
467
MATERIALS AND METHODS order of the operations involved first placement
of the bed, followed by inoculation with
The experiment was carried out in the spring of earthworms (Eisenia foetida) and then
2017 within the vegetable growing sector of the repeatedly covering with thin layers of 10-15
University of Agronomic Sciences and cm of compost.
Veterinary Medicine of Bucharest, and it In time, a stratified stratum is created, with the
assumes the installation of a monofactorial finished product at the bottom (vermicompost),
experimental scheme with the following partially consumed in the middle and fresh
variants: V1 - unfertilized (100% peat); V2 - topped food.
seedlings fertilized in the substrate with 10%
vermicompost (90% peat); V3 - seedlings RESULTS AND DISCUSSIONS
fertilized in the substrate with 20%
vermicompost (80% peat); V4 - foliar fertilized The analysis of the seed growth results is
seedlings with filter macerated vermicompost presented in Table 1.
in 20% concentration and V5 - foliar fertilized It is highlighted that the plants achieved the
seedlings with 30% filtered macerated best and balanced growth in V3, where all the
vermicompost. In order to produce the analyzed indicators recorded the highest values:
seedlings were made directly in the box in the 6.2 formed leaves; 28.2 cm plant height; 22.8
production department of the seedlings in the cm root length and 0.21 leaf frequency,
Hortinvest greenhouses on the 14th of February followed closely by V5 and V4 that had
2017, the boxes being filled with the Klasman balanced growths. At the opposite end, V1,
TS3 peat. Mass uprising occurred very quickly, unfertilized, achieved the weakest growth, all
9 days after the date of sowing, February 23, the indicators analyzed being lower than the
2017. Sowing was done on March 2nd in 330 cc other variants.
plastic pot filled with Klasman TS3 ±
vermicompost 10% and 20% (V2 and V3) peat, Table 1. Growth of eggplant seedlings at the end of the
according to the experimental protocol. Care experimentation period
work has been applied uniformly to ensure Var. NFZ HPA HR NFZ/HPA
optimal growth and development conditions, (cm) (cm) (no./cm)
automated, computer-assisted: Temperature To V1 5.8 22.6 13.6 0.25
= 20-28oC; substrate humidity 55-65%; V2 6.0 25.8 18.2 0.23
ventilation of the space and maintaining the V3 6.2 28.2 22.8 0.21
V4 5.6 25.6 21.8 0.21
correct lighting regime.
V5 5.8 29.2 16.8 0.19
In order to evaluate the influence of
vermicompost on the growth and development
As can be seen from Figures 1 and 2,
of seedlings, observations and measurements
vermicompost fertilization has influenced at
were made at one moment only, at the end of
least significantly the height of the aerial part
the experimentation period, when the seedlings
(R2HPA = 0.639) and the seed growth balance,
were good for planting, April 10, 2017, the
respectively the leaf frequency (R2 = 0.9423).
seedlings age being of 52 days.
By direct observations and measurements we
determined: plant height (HPA); root length R² = 0,639
(HR); the number of true formed leaves (NFZ); 35
30 25.8
28.2
25.6
29.2
leaf frequency (NFZ / HPA); collet diameter 25
22.6
20
(collet Ø); mass of aerial vegetative apparatus 15
(MPA); mass (MR); the volume of roots (VR); 10
5
and the total mass of seedlings (Mtot). 0
The vermicompost used in the experience was V1 V2 V3 V4 V5
produced at the Vegetable Research- HPA (cm) Linear (HPA (cm))
Development Station Buzau, using the system
of continuously fed beds. Beds are elongated,
Figure 1. Influence of fertilization with vermicompost on
in a rick style, with a height of 50 cm. The the growth of eggplant
468
Results on the development of eggplants, 25 R² = 0,83
evaluated by means of mass, volume and 20
diameter indicators, are presented in Table 2 15
20,2 19,6 19,6 18,6
and Figure 3. 10 12,4
5
0
R² = 0,9423 V1 V2 V3 V4 V5
0.25
0.23
0.21 0.21 Mtot (g) Poly. (Mtot (g))
0.19
Figure 3. Influence of vermicompost fertilization on the

total weight of eggplant seedlings
V1 V2 V3 V4 V5
The indicator showing the weakest difference is
the volume of the root system where, for V2
Frecvenţa frz. Linear (Frecvenţa frz.)
and V3, it recorded 9.8 cm3 and for V4 and V5
Figure 2. Influence of fertilization with vermicompost on
9 cm3. The other indicators showed much
the frequency of leaves in eggplants stronger tones, presented and discussed
throughout the paper.
Looking at the overall experience, from the
Results on the development of eggplants,
point of view of the growth and development of
evaluated by means of mass, volume and
the eggplants, V3 is highlighted, fertilized in
diameter indicators, are presented in Table 2
substrate with 20% vermicompost, having the
and Figure 3.
best and balanced growth and development of
Analysis of the results again reveals V3 as the
seedlings. Diametrically opposed, the least
most balanced; In general, the indicators have
favourable variant is the unfertilized control
recorded superior values (10.4 g aerial parts
variant, which recorded the most unbalanced
weight, 9.8 cm3 root volume and 6.2 mm collet
growth and development of seedlings.
diameter), or equal to the other variants: root
mass 9.3 g and total mass 19.6 g. This is
CONCLUSIONS
followed by V2 and V4, variants that have
values close to V3 and superior to the other In carefully analysing the results and
variants. V1 has been shown to be the weakest interpretations made, we can say that substrate
variant in terms of accumulation compared to fertilization with 20% vermicompost,
other variants. determined the obtining of the best seedlings.
Very good results in the quality of seedlings
Table 2. Development of seedlings at the end of the
experimentation period have been obtained in the case of V2 and V5.
We note that fertilization in the substrate has
Var.
MPA MR Mtot VR Ø collet provided a better growth and development of
(g) (g) (g) (cm3) (mm)
eggplants seedlings, compared to the foliar
V1 7.2 5.2 12.4 5.2 5 application of filtered macerate vermicompost.
V2 8.4 11.8 20.2 9.8 5 As a result of the vermicompost fertilization
V3 10.4 9.2 19.6 9.8 6.2 program of eggplants, their age has decreased
V4 7.2 12.4 19.6 9 5 from 75 - 80 days (Ciofu et al., 2003; Dobrin
V5 9.4 9.2 18.6 9 5.2 2016), to 52 days, which makes it possible to
From Figure 3 it can be seen that fertilization achieve important economies with the warming
with vermicompost has a decisive and complex of the seedlings production areas, because the
influence on the total mass accumulations of sowing can be done almost 1 month later.
plants (R2 = 0.83). Therefore, we recommend to the seedlings
The indicator showing the weakest differences producers, using the three fertilization
is the volume of the root system, where for V2 formulas, respectively 20% and 10%
and V3 it recorded 9.8 cm3, and for V4 and V5 vermicompost in the substrate and foliar
9 cm3. The other indicators showed much fertilization with 30% filter maceration
stronger nuances. vermicompost for the production of eggplants.
469
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Ciofu R., Stan N., Popescu V., Chilom P., Apahidean S., index. International Journal of Vegetable Science 16:
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470
EFFECT OF THE USE OF NEW METHODS FOR

THE REMEDIATION OF OIL POLLUTED SOIL
Elena Maria DRĂGHICI1, Virgil SCARLAT 2, Maria PELE2, Elena DOBRIN 1,
Gabi-Mirela MATEI3, Sorin MATEI 3
1
Department of Horticulture, 59 Mărăşti Blvd,
2
University of Agronomic Sciences and Veterinary Medicine of Bucharest, Faculty of
Biotechologies, Department of Biotechnologies, 59 Mărăşti Blvd, District 1, Bucharest, Romania
3
National R&D Institute for Soil Science, Agrochemistry and Environment, Research Department
of Soil Science, Agrochemistry and Environment, 61 Mărăşti Blvd., District 1, Bucharest, Romania
Abstract
The present study performed the investigation of new environmental friendly methods to remediate polluted soil with
crude-oil. The experiment was carried out in greenhouse, in controlled condition using polluted soil and perlite, in
different percent. Also, we used five organic fertilizers (Vermiplant, Amalgerol, Poco, Iguana and Formulex). Based on
the data obtained under controlled conditions, the best results were obtained in variants where the percentage of perlite
used in the soil mix was 25%. The 50% and 75% perlite mix with polluted soil quantities used in the blend did not bring
significant differences in plant growth. The research on the influence of remedial measures of the polluted soil on main
growing parameters revealed significant differences on physical and chemical properties of the soil, on plant growth,
biomass production and plant biochemical composition before and after the treatment. The proposed technology
effectively recovered soil properties and plant growth. Microbial populations included 6-15 bacterial strains belonging
to Pseudomonas, Bacillaceae and actinomycetes and 5-10 fungal species. Treatments, especially when applied on soil
diluted with perlite stimulated species diversity increasing and pathogen inhibition.
Key words: bean, bioremediation, microbial populations, pollution, perlite.
INTRODUCTION there is the phenomenon of expansion and

breaking of the granules. A granular material is
The technology for remediation of soil obtained very easily and porous, with various
contaminated with petroleum residues or granules, from 0 to 6 mm.
residual salts, which is the subject of this Plants germinate and grow in environments
scientific work, is concerned with the use of where water, mineral elements and air are
expanded perlite as a constitutive element of available. Oil pollution is a major risk for the
soil quality improvement recipe. biodiversity of the land due to the fact that both
The expanded perlite is a granular, light and crop and bacterial microflora are affected.
porous product based on chemically stable Oil reduces soil fertility so nutrients become
(SiO2, Al2O3, K2O, Na2O, Fe2O3, CaO, MgO, inaccessible for plants. Oil has a higher density
TiO2) with density ranging from 40 to 250 than water and displaces air and water between
kg/m3. The raw material is the perlitic rock, of soil particles, making it difficult for mineral
volcanic origin, generated by the lava solidified absorption and triggering anaerobic
in water. Through this process, the molecular metabolism. Soil properties determine the rate
structure of the rock remained chemically of absorption and plant growth, directly
bound water. The presence of water molecules affecting crop production.
causes the explosion of rock granules brought Bioremediation and natural rehabilitation occur
to temperatures of 800-1000oC. Due to the over a long time, and as such in the
semiplastic state, where the granules of rock rehabilitation of these soils, the scientific
are found at the temperature of the furnace, community is involved in finding optimal and
471
rapid solutions (Malschi, 2015; Baek et al., manganese and amino acids. POCO is a totally
2004). Aromatic hydrocarbons found in natural product with pH: 8.5-9.5 that contained:
petroleum are large complex high-persistence Calcium 0.04-0.05%; Iodine: 6.30-12.70 mg/l;
molecules that typically require powerful Magnesium 0,50-0,80 mg/l; Nitrogen (N):
reagents to annihilate their action. 0.025-0.038 mg/l; Potassium: 0.50-0.64%;
Compared with biological methods, physical Sodium (Na): 0.088-0.120%; Sulfur 0.028-
and chemical methods may produce secondary 0.050%. Iguana is a organic product with 4%
pollution to reclaim oil contaminated soil (Zhan nitrogen, 3% phosphorus, 6% potassium.
et al., 2017). Formulex is also an organic product that
contains: N 2.40%; P2O5 0.85%; K2O 3.36%;
MATERIALS AND METHODS CaO 1.85%; B 0.0108% and microelements.
Experiments were performed in the greenhouse RESULTS AND DISCUSSIONS

conditions. We used the cultivar Unidor of
bean characterized through determined hight, Data obtained from bean plants grown under
vegetative period of 79 days and resistences at controlled conditions showed that the best
anthracnose, Colletotrichum lindemuthianum; results were obtained in variants where the
BCMV-VMT and PSP-Pseudomonas syringae percentage of perlite used in the soil mix was
pv. phaseolicola. 25%. The 50% or 75% perlite quantities used in
The polluted soil was collected from Icoana the blend did not bring significant differences
commune, Olt district, situated in the SV part in plant growth (Figure 1).
of Romania.
We performed soil mixtures with perlite in
varying proportions, and fertilization was
performed with Amalgerol, Vermiplant, Poco,
Iguana and Formulex in a 2%. The experi-
mental variants are presented in Table 1.
Table 1.The experimental variants
Variant V1 V2 V3 V4 Figure 1. Experimental variants
100% 75% 50% 25%
polluted polluted soil polluted soil polluted
soil +25% +50% soil +75% In the case of polluted soil (V1), the biomass of
perlite perlite perlite bean plants was higher in the variant that we
Control V1/1 V2/1 V3/1 V4/1
Amalgerol V1/2 V2/2 V3/2 V4/2
administered the Vermiplant product (176.25 g
Vermiplant V1/3 V2/3 V3/3 V4/3 V1/3). This was with 16.75 g higher than the
Poco V1/4 V2/4 V3/4 V4/4 control variant (159.5 g V1/1)). The variant we
Iguana V1/5 V2/5 V3/5 V4/5 improved the soil with 25% perlite and
Formulex V1/6 V2/6 V3/6 V4/6
fertilized with Vermiplant produced an average
plant biomass of 201.67 g, with 32.67 g more
We determined: the amount of water required
than the control variant (Figure 2).
to soak up the soil; the amount of water
consumed by plants (throughout the
experiment); growth dynamics of plants; the 210
200
total vegetative mass resulting; distribution of 190 176,25
Biomass (g)
180 167,67 164,50 160,67 161,33

the root system (at the end of growth). 170 159,50
160
The caracteristic of fertilizers used in 150
140
experiment. Amalgerol is a product with that 130
contain natural oils with benefic effects on the
plant regarding increases the root mass of
V1 control Polluted soil
plants and the absorption of nutrients. V2- 75% polluted soil +25%perlit
VermiPLANT is a totally natural product V3 50% polluted soil +50% perlit
V4- 25% polluted soil+75% perlit
obtained from earthworms that contains
microelements such as barium, zinc, iron, Figure 2. Influence of substrate on plant biomass
472
Statistically analyzing the obtained data, we 175
found that, in the case of 25% perlite soil 170
improvement and organic fertilizers, we 165
Biomass (g)
obtained increases in the mass of plants, in all 160
experimental variants, the differences being 155
150
very positive at V2 (Table 2). 145 y = -5,9833x + 173,5
R² = 0,393
140
Table 2. Total biomass of bean plants 135
0,5 1,5 2,5 3,5 4,5
Variants V1 V2 V3 V4 variants
Control 159.50 169.00*** 164.67** 141.00 ooo Figure 3 Influence of soil improvement with perlite
Differences 100.00 105.96 103.24 88.40 at the control variant on plant biomass
DL5%= 2.690 DL5% in % = 1.6865
DL1% = 4.080 DL1% in % = 2.5580
DL0.1%= 6.50 DL0.1% in %= 4.0752
Amalgerol 167.67 196.33*** 178.00*** 150.00 ooo
Differences 100.00 117.09 106.16 89.46
DL5% = 2.050 DL5% in % = 1.2226;
Biomass (g)
DL1% = 3.100 DL1% in % = 1.8489;
DL0.1% =4.950 DL0.1% in %= 2.9522
Vermiplant 176.25 201.67*** 150.67 ooo 142.50 ooo
Differences 100.00 114.42 85.49 80.85
DL5% = 0.110 DL5% in % = 0.0624;
DL1%= 0.170; DL1% in % = 0.0965;
DL0.1% = 0.280 DL0.1% in %= 0.1589
Poco 164.50 184.00*** 157.33 oo 143.25 ooo variants
Differences 100.00 111.85 95.64 87.08
DL5% = 4.230 DL5% in % = 2.5714 Figure 4. Influence of soil improvement with perlite at
DL1% = 6.400 DL1% in % = 3.8906 variant fertilized with Amalgerol on plant biomass
DL0.1% = 10.190 DL0.1% in %= 6.1945
Iguana 147.56 199.00 154.20 138.17
Differences 100.00 134.86 ** 104.50 93.64
DL5% = 21.350 DL5% in % = 14.4690;
DL1% = 32.300 DL1% in % = 21.8899
Formulex 161.33 191.25 140.00 166.00
Differences 100.00 118.55*** 86.78 ooo 102.89 **
DL5% =1.970 DL5% in % = 1.2211;
DL1% = 2.980 DL1% in % = 1.8471;
DL0.1% = 4.760 DL0.1% in %= 2.9505
V1 - contaminated soil
V2 - contaminated soil 75%+25% perlite
Figure 5. Influence of soil improvement with perlite at
In the case of applying a higher amount of variant fertilized with Vermiplant on plant biomass
perlite to the soil by applying the fertilizer
dose, we obtained statistically significant
negative results in most of the variants except
for the variant to which the Formulex product
was applied.
The correlations between the applied product
and the perlite-polluted soil mixture, in varying
percentages (Figures 3-8), led to the conclusion
that there was a significant correlation with the
Vermiplant product (R2=0.5386) (Figure 5).
For Formulex, the relationship was
insignificant (R² = 0.0522) (Figure 8). Figure 6. Influence of soil improvement with perlite at
variant fertilized with Poco on plant biomass
473
183
173
Biomass (g)
163 y = -5,3733x + 176,28
R² = 0,6048
153
143
133
-0,5 1,5 3,5 5,5 7,5
variants
Figure 7. Influence of soil improvement with perlite at Figure 11. Influence of fertilizers on polluted soil
variant fertilized with Iguana on plant biomass ameliorated with perlite in percent of 50%
163
Biomass (g)
153 y = 2,5786x + 137,79
R² = 0,2246
143
133
0 2 4 6 8
variants
Figure 12. Influence of fertilizers on polluted soil

Figure 8. Influence of soil improvement with perlite at
ameliorated with perlite in percent of 75%
variant fertilized with Formulex on plant biomass
Analyzing the influence of the above men-

tioned fertilizers on the perlite-soil combi-
nations (Figures 9-12), we estimate that for the
control variant (polluted soil), the influence of
fertilization application showed insignificant
relationship (R²=0.0405) (Figure 9).
180
175
Biomass (g)
y = -0,6738x + 167,34
170
R² = 0,0405
165 Figure 13 Aspect of bean plants in experiment
160
5,6
155
0,5 2,5 4,5 6,5 5,1 5,2 5,2
variants 4,8
4,6
Figure 9. Influence of fertilizers on polluted soil 4,1

Dry matter (g)
4
3,8
3,6
205 3,1 3,1 3 3
200 2,7
2,6 2,5
195 2,4 2,4
2,2 2,2 2,2
Biomass (g)
190 2,1 2 2,1 2 2,1

y = 2,9024x + 180,05 1,8
185
R² = 0,2004 1,6 1,7
180 1,4
1,1 1,2
175
170
165
0,5 2,5 4,5 6,5 V1 polutted soil
variants V2 75% polutted soil+25%perlite
V3 50% spolutted soil+50% perlite
V3- 25% polutted soil+75% perlite
Figure 10. Influence of fertilizers on polluted soil
ameliorated with perlite in percent of 25% Figure 14. Dry matter content of the roots
474
Analyzing the aspect of plants (Figure13) and Aspergillus, accompanied by antagonists
dry matter content of the roots, we found that Trichoderma, Paecilomyces and other species
the highest values were recorded in soil with high ability for enzymatic degradation of
variants improved by 50% perlite, and fertilized various organic substrates were identified with
with Vermiplant (5.2 g) and the variant high frequency in soil with perlite and
improved with 75% perlite and treated with stimulating treatments (Figure 17).
Poco (Figure 14).
The highest dry matter content of the leaves
was recorded in the 25% perlite-enhanced soil
variant, and fertilized with Iguana (Figure15).
16,3 16,4
14,3
Dry matter (g)
12,3
12 11,8
11,2 11
10,8
10,3 10,1 10,2 10,2 10,4
9,8 10
9,4 9,4
9 9
8,8
8,3 8,2 Figure16. Aspects of the bacteria developed in 75%
7,5 7,6 polluted soil + 25% perlite and Iguana (V2/5)
6,8
6,3
5,2
5
4,3
V1 polutted soil
V2 75% polutted soil+25%perlite
V3 50% spolutted soil+50% perlite
V3- 25% polutted soil+75% perlite
Figure 15. Dry matter content of the leaves
Microbial populations in experimental variants

were represented by 6 to15 bacterial species
including actinomycetes belonging to 1-4 Figure17. Aspects of the fungi developed in 25%
Series. The stimulation of species diversity was polluted soil + 75% perlite (V4/2) and Amalgerol
noticed and ecologic ameliorative treatments
were added to soil, especially in variants mixed Results of the present study confirm our
with various proportions of perlite. previous research that showed the involvement
In many variants, Pseudomonas fluorescens of pseudomonads (Matei et al., 2007; Scarlat et
was dominant, followed by different species of al., 2015) and Aspergillus species in
Bacillaceae (Figure 16). Fungal microflora hydrocarbon degradative processes.
included communities of 5 to maximum 10 Other authors reported beneficial effect of
species with variation as a function of bioremediation technologies including bacterial
experimental factors (substrate type and the and fungal biodegraders with stimulating
nature of stimulating treatment applied). An treatments for soil conditions improvement and
inhibition of Fusarium pathogenic species as plant growth and enhancing (Potin et al., 2004;
compared with control was evidenced in Verma et al., 2006).
polluted soil under the influence of various
stimulate treatments and the dominance of CONCLUSIONS
cellulolytic and hydrocarbon degrading species
Cunninghamella elegans. Species of genus Application of organic fertilizers has led to the
increase of plant mass in all variants. When
475
Vermiplant was applied (V2/3), the highest Research, CNDI-UEFISCDI, project number
plant mass was 201.67 g compared with the rest PN-III-P2-2.1_PTE-2016-0084, (40-PTE),
of the variants. acronym ECOREMTEH.
There were direct relationships between the
polluted soil mixture (75% polluted soil and REFERENCES
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Baek K.H., Kim H.S., Oh H.M., Yoon B.D., Kim J., Lee
was result that the variants cultivated on a I.S., 2004, Effect of crude oil, oil components and
mixture of 50% polluted soil mixed with 50% bioremediation on plant growth, Journal of
perlite with application of Amalgerol (V3/2) environmental science and health, Part A -
and also in the 25% polluted soil mixed with Toxic/Hazardous Substances & environmental
75% perlite with application of Poco products ngineering vol. A39 (9): 2465-2472.
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(V4/4), the dry matter content was the highest Bioremediation of Petroleum Contaminated Soil.
(5.2 g for both variants). Journal of Yangtze University (Natural Science
The highest dry matter content at the leaves Edition), 13: 52-56.
was recorded in the fertilized variant with the Matei S., Matei G.M., Mocanu A., 2007. The dynamic
Iguana product, plants grown on the substrate modifications of Diesel oil fractions in a Chromic
Luvisol under remedial measures, Știința Solului, 41
with polluted soil (75%) and perlite (25%) (1): 88-100.
(V2/5). Scarlat V., Pele M., Drăghici E.M., 2015. Evaluation the
Microbial populations included 6-15 bacterial ability of the fungus Aspergillus to remove oil from
species dominated by Pseudomonas, contaminated soil, Scientific Papers, Series B,
Bacillaceae and actinomycetes and 5-10 fungal Horticulture, 59: 463-466.
Potin O., Rafin C., Veignie E., 2004. Bioremediation of
species. an aged polyciclic aromatic hydrocarbons (PAHs)-
Treatments, especially when applied on soil contaminated soil by filamentous fungi isolated from
diluted with perlite stimulated species diversity the soil, International Bioremediation and
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This work was supported by a grant of the
Romanian National Authority for Scientific
476
THE INFLUENCE OF ALTERNATIVE TECHNOLOGICAL SEQUENCES

ON THE QUALITY OF MELON PRODUCTION
Rodica SOARE1, Maria DINU2, Cristina BĂBEANU3, Marin SOARE1

1
Faculty of Agriculture, University of Craiova, 19 Libertăţii Street, 200583, Craiova, Romania
2
Faculty of Horticulture, University of Craiova, 13 A.I. Cuza Street, Craiova, Romania
3
Faculty of Sciences, University of Craiova, 13 A.I. Cuza Street, Craiova, Romania
Abstract
At present, there is a growing consumer interest in high value-added vegetable products. It is therefore necessary to
limit the use of fertilizers and synthetic chemical pesticides because of the harmful effect on the environment. In this
regard, a melon culture was established in the polyethylene tunnels to which Lignohumat biostimulator product was
applied in vegetation as an alternative for chemical foliar fertilization. It is a humic product, granulated with
microelements and with the effect of growth and protection against stress factors. Three treatments were applied in
three doses of fertilization. The specific of variants was: V1 (Control)-unfertilized; V2-fertilized with 100 g/ha; V3-
fertilized with 150 g/ha; V4-fertilized with 200 g/ha. The determinations focused on the quality of the production. The
200 g/ha Lignohumat dose influenced the quality attributes and the dose of 100 g/ha Lignohumat the antioxidant
activity of fruit. Thus, was recorded a SUS content of 8.05%, SUT of 12.29%, reducing sugars of 3.73%, vitamin C of
29.4 mg/100 g, TP of 42.03 mg GAE/100 g, carotene of 20.98 mg/100 g f.w. and antioxidant activity of 184.44
μMTE/100 g.
Key words: Cucumis melo, carotene, organic fertilization.
INTRODUCTION is essential for enzyme activation, protein

synthesis, for photosynthesis and mediates
In Romania, the watermelon (Cucumis melo L.) osmoregulation during cell expansion and for
is highly appreciated and annually cultivated stomatal movements and tropisms (Silva et al.,
especially in South and South-East Romania, 2007). Nutritional imbalances, particularly
and in recent years there has been an increase inadequate K supply, often contribute
in protected crops. Melon fruits are consumed significantly to yields and low crop quality
at physiological maturity, fresh or mixed with (Jifon, 2011).
other fruits, such as dessert salads and also In recent years, bio-fertilizers based on humic
processed in the form of jams, etc. Cantaloupe acids are increasingly used, preparations that
is a very good source of vitamins A, C and β- reduce the negative effects of chemical
carotene and can be used as juice fruit fertilizers on the environment. Fertilizers with
(Mohamed and Maha-Mohamed, 2016). Also, humic substances can be used on different
the melon seeds are a good source of natural types of soil, as well as in technologies for
antioxidants with significant biological improving degraded or contaminated soils and
functions and may serve as food ingredients are efficient for many cultures (Sîrbu et al.,
(Zeb., 2015). By its composition, melon can be 2016). Humic acids replace the requirements of
a rich source of many nutrients related to other fertilizers, increase productivity, improve
human health (Lester, 2008). soil drainage and establish a favorable
It is very important to find the optimal environment for the development of soil
fertilization dose as well as the appropriate microorganisms (Salman et al., 2005). In
combinations of fertilizersfor plant health, high addition, they can induce changes in plant
productivity and fruit quality (Dinu et al., primary and secondary metabolism related to
2017; Bouzo and Cortez, 2015). Potassium is abiotic stress tolerance which collectively
an important macronutrient and the most modulate plant growth (Canellas et al., 2015).
abundant cation of the superior plants. Also, K+ The application of humic acids has been
477
reported by many authors with a positive effect methanol, intensely agitated for 10 minutes
on tomato plant growth and production (Dinu et using avortex. The extraction of phenolic
al., 2015), the growth of pepper production compounds was carried out by keeping the
(Karakurt et al., 2009), germination of tomato mixture in an ultrasonic bath for 70 minutes.
seeds (Dinu et al., 2013), growth and The mixture was filtered, and to the residue
development of Spathiphyllum wallisii (Manda solid was added 5 mL of 80% methanol and the
et al., 2014). Thus, the application of extraction procedure repeated. Obtained the
biostimulators can be considered a good two extracts were combined and analyzed using
strategy of vegetative and generative growth of protocols specific work to determine the total
plants, a high nutritional value of production polyphenols and antioxidant capacity.
and low impact on the environment Dry matter content was determined
(Becherescu et al., 2016; Dobrin et al., 2016; gravimetrically at 105°C to constant weight
Naidu et al., 2016; Parađiković et al., 2011). (ISO751: 2000) (%) and the soluble solids
This study aims to investigate the effect of content (SSC) (%) was determined using a
Lignohumat doses, biological fertilizer based digital refractometer (KrussOptronic DR 301-
on humic acids, on the quality of melon fruit. 95) at 20°C.
The acidity was determined by titration with
MATERIALS AND METHODS 0.1 N sodium hydroxide (NaOH) and expressed
as (%) citric acid and reducing sugars (%) were
The experience was placed in the polyethylene extracted in distilled water (1:50 w/V) and
tunnels of the Faculty of Horticulture and assayed colorimetric with 3.5-dinitrosalicylic
Faculty of Agriculture, University of Craiova, acid using glucose as standard.
Romania, in randomized blocks, in three The determination of vitamin C content
repetitions, in 2015-2016. The biological A sample of 5-10 g of tomatoes, previously
material was Ananas cultivar (Cucumis melo ground with quartz sand has been put into a 100
var. reticulatus) oval fruits with a pineapple ml balloon by using a solution of 2%
scent and orange skin with reticulated hydrochloric acid. It has been stirred and after
appearance. Date of planting to the sedimenting it has been filtered into a dry glass.
polyethylene tunnels was in the first decade of A 10 ml aliquot has been passed into a
May while plant density was 3.2 plants m/2. Berzelius glass, to which 30 ml of distilled
The specific of the variants was: V1 (Control) - water; 5 ml of 1% potassium iodate and 1 ml
unfertilized; V2- fertilized with 100 g/ha solution of starch have been added. It has been
Lignohumate; V3-fertilized with 150 g/ha then titrated with potassium iodate N/250
Lignohumate; V4- fertilized with 200 g/ha stirred until becoming bluish (Chowdhury,
Lignohumate. This is a foliar humic 2004).
biostimulator, containing microelements in the The determination of total carotenoids
form of chelates having the highest dose of The weighed samples, having been put
Potassium Humate 850 g/kg. separately in 95% in acetone (50 ml for each
Lignohumat organic fertilizer was applied gram), were homogenized with Braun MR 404
foliarly in three treatments at 10 days interval Plus for one minute. The homogenate was
and the culture technology was that specific to filtered and was centrifuged using the Hettich
melons grown in polyethylene tunnels. During Universal 320/320R centrifuge at 2500 rpm for
the growing season, it also pursued the ten minutes. The supernatant was separated and
application of some products to combat weeds, the absorbances were read at 400-700 nm on
diseases and pests with low environmental Cary 50 spectrophotometer.
impact products for sustainable agriculture. The total phenols (TP) in melon fruit was
Thus, at the basic fertilization was applied estimated by the method proposed by Mallick
20 t/ha compost. and Singh (1980).
Samples preparation The capacity of extracts to reduce the radical
A quantity of100g of each sample was broken 2.2-diphenyl-1-picrylhydrazyl was assessed
up in blenduire to mix. 1 gram of homogenized colorimetric. 2 mL of 0.075 mM DPPH
sample (pulp) was mixed with 10 mL 80% solution in ethanol was mixed with 0.1 mL
478
extract and vortexed thoroughly. The Fruit quality is determined by complex
absorbance of the remaining DPPH radicals networks of metabolic pathways developed
was measured at 517 nm. The results were during fruit ripening. melon (Cucumis melo L.)
expressed as μMTrolox equivalents (TE)/100 g. comprises a broad array of genotypes, in which
The ABTS radical cation scavenging activity of the fruit accumulate soluble sugars, organic
the methanolic extract was assessed acids, secondary metabolites of pigmentation
colorimetric. ABTS was dissolved in water to a and aroma volatiles to varying levels. In
7 mmol/l concentration. ABTS radical cation addition, Cucumis melo includes fruit with
was produced by reacting ABTS stock solution different biochemical accumulations in
with 2.45 mmol/l potassium persulfate and ripening physiology, levels of ascorbic acid,
allowing the mixture to stand in the dark at additional secondary metabolites and other
room temperature for 12-16 h before use. The components of quality (Katzir et al., 2008).
ABTS radical cation solution was diluted with In the present paper, the total dry substance
ethanol to an absorbance of 0.70 at 734 nm. 0.1 (TDS) of melon fruit recorded upward values,
ml of sample extract was mixed with 2.9 ml of along with increasing the fertilization dose,
diluted ABTS radical cation solution. After from 7.59% at the control variant to 12.29% at
reaction at room temperature for 6 min., the the fertilized variant with Lignohumat 200
absorbance at 734 nm was measured. The g/ha. Regarding the soluble solids content
Trolox calibration curve was plotted as a (SSC), it also recorded upward values with the
function of the percentage of ABTS radical fertilization dose (Table 1). The values found
cation scavenging activity. The final results are similar to those reported by Salman et al.
were expressed as μMTrolox equivalents (2005), on the growth of total dry substance
(TE)/100 g fw (Soare et al., 2016). and the soluble solids content with the increase
Statistical analysis in humate acid potassium, in watermelon,
The data recorded were statistically processed Aswan hybrid. Some authors have concluded
by using the analysis of the variance (ANOVA) that in cucumber plants treated with nitrogen-
with a significance level of p<0.05 by based biofertilizer at higher doses, it resulted
Duncan’s multiple range test. Also, were total soluble solids higher compared to lower
carried correlations between the analysed doses (AbdAlla et al., 2009; Oliveira et al.,
characters (Pearson’s correlation analysis). 2003)
The effect of the culture system on titratable
RESULTS AND DISCUSSIONS acidity may actually be an important factor in
the quality fruit. The titratable acidity varied in
Inappropriate fertilizations with N, P and K the four studied variants depending on the
cause nutritional imbalances, also inducing fertilization dose. The highest values were
deficiencies of calcium and boron, forming recorded in variant which had the highest
fruits with deformations, abnormal color and fertilization dose, Lignohumat 200 g/ha.
low storage capacity (Silva et al., 2007). Different concentrations of humic acid-based
Precise estimates of quantities of nutrients for products can influence the acidity level of
crops as well as application in critical times are edible cabbage (Soare et al., 2017) or from
essential for increasing production and quality pepper (Aminifard et al., 2012).
while protecting the environment.
Table 1. Analysis of some quality attributes of melon fruits
Variant TDS (%) (SSC) Acidity (citric Reducing Vitamin C
(%ºBrix) acid %) sugars (%) (mg/100g f.w.)
V1(Control)- unfertilized 7.59b 5.55c 0.123ab 3.15b 21.2b
V2- fertilized with Lignohumat 100 g/ha 7.94b 6.25bc 0.112bc 1.81c 29.4a
V3- fertilized with Lignohumat 150 g/ha 7.86b 6.95b 0.074c 2.84b 17.9c
V4- fertilized with Lignohumat 200 g/ha 12.29a 8.05a 0.156a 3.73a 22.8b
LSDİ0.05 0.82 0.69 0.04 0.50 1.85
Different letters within the same row indicated significant differences (P < 0.05) among variants
479
Reducing sugars, an important parameter for 38.1 mg/100 g fresh weight C vitamin.
determining fruit quality of melon, recorded According to some authors, the vitamin C
fluctuating values depending on the variant. content of vegetables can vary depending on
The content of reducing sugars in fruits treated the intensity of light, temperature, humidity,
with Lignohumat 200 g/ha was signicantly pollution (Dinu et al., 2016).
higher (3.73%) in comparison with fruits from Total polyphenols in the present study recorded
the control variant. The increase of reducing significant differences in the variant treated
sugars under the influence of products based on with 100 g/ha of Lignohumat, 42.03 mg
humic acids was also recorded in autumn white GAE/100 g f.w., after which it decreased with
cabbage (Soare et al., 2017). Lester (2008) the increase of the fertilization dose.
claims that the accumulation of sucrose in Salandanan et al. (2009) noted a high
melon depends on several factors and is variability of the total polyphenols in some
maximal in the final stages of fruit maturation. melon varieties, influenced by yearly
The antioxidant potential was evaluated by environmental effects and production system
total polyphenols and vitamin C contents, (conventional and organic), from 40.5 to 71.7
carotenes, and antioxidant capacity. The mg GAE/100 g f.w. and Selale et al. (2012) in
presence of phenolic compounds, such as their paper, at 42 melon lines and cultivars of
flavonoids, phenolic acids and anthocyanins, different types, reported that total phenolic
besides the vitamins C, E and carotenoids, content in the melon ranged from 118.5 to
contribute to the beneficial effects on human 357.8 mg GAE/kg−1.
health (Podsedek, 2007). The highest value of antioxidant activity was
Concerning the vitamin C, in the present study, also recorded in the fertilized variants
it was registered a variation amplitude of 17.9 compared to the control. Astfel, the ABTS
mg/100g f.w. (Lignohumat 150 g/ha) to 29.4 value ranging between 88.76 μMTE/100 g
mg/100g f.w. (Lignohumat 100 g/ha). In the (unfertilized) and 150.72 μMTE/100 g
variant fertilized with Lignohumat 200 g/ha, (fertilized with Lignohumat 100 g/ha).
vitamin C was 22.8 mg/100g f.w. There is a Increasing doses of Lignohumat did not
high variability of ascorbic acid (active form of influence the antioxidant capacity, probably
vitamin C) influenced by Lignohumat due to the higher content of Potassium Humate
concentration. Similar results have been and microelements. The results are similar to
reported Salandanan et al. (2009) in a study those reproted by Salandanan et al. (2009) at
regarding the antioxidant properties and quality the organically and conventionally fertilized
attributes or 10 melon (Cucumis melo L.) melon cultivars values of antioxidant capacity
cultivars grown under conventional and organic from 49 to 220.5 μmol TEAC/100 g f.w.
conditions, recorded a variation from 16.2 to
Table 2. Biochemical determinations of melon fruit
Total phenolics Total carotene Antioxidant activity
Variant (mg GAE/100 g f.w.) (mg/100 g f.w.) ABTS DPPH
μMTE/100 g μMTE/100 g
V1(Control)- unfertilized 30.1b 17.38d 88.76c 91.46d
V2- fertilized with Lignohumat 100 g/ha 42.03a 20.98c 150.72a 188.44a
V3- fertilized with Lignohumat 150 g/ha 23.91c 26.28a 113.29b 136.39b
V4- fertilized with Lignohumat 200 g/ha 23.24c 25.1b 89.89c 113.13c
LSDİ0.05 3.36 2.72 5.79 4.50
Different letters within the same row indicated significant differences (P < 0.05) among variants
The same, DPPH radical scavenging activity g/ha). The unfertilized variant recorded the
decreased as the fertilization dose increased lowest value of 91.46μMTE/100 g.
from 188.44 μMTE/100 g, (Lignohumat 100 Parađiković et al. (2011) have claimed that
g/ha) to 113.13 μMTE/100 g (Lignohumat 200 biostimulators improve antioxidant activity,
vitamin C content and phenols, compared to
480
untreated fruits. Some authors said that fruits with high antioxidant potential
cultivars selection, climate conditions, fertilizer (Wulandari et al., 2016).
types, culture system (conventional or organic) Regarding the correlation coefficient between
may influence the antioxidant activity of some quality characters, positive correlation
vegetables (Apahidean, 2017; Dinu et al., 2016; coefficient values were obtained between
Salandanan et al., 2009; Salman et al., 2005). ABTS and total polyphenols (0.935), between
Other authors state that irrespective of the DPPH and total polyphenols (0.962) and
analysed index, variants organically fertilized between DPPH and ABTS (0.996) (Table 3).
are significantly superior to the control variant These results agreed with those reported by
(Draghici et al., 2016). Also, the maturing stage Selale (2012) which showed significant
is an important factor in the antioxidant correlation between antioxidant capacity and
properties of melon, and the cultivar must phenolic content.
determine the appropriate harvesting time for
Table 3. Coefficients of correlation (r) between some quality characters of melon
Specification Total phenolics Total carotene ABTS DPPH Reducing Vitamin C
(mg GAE/100 g) (mg/100 g f w ) μMTE/100 g μMTE/100 g sugars (%) (mg/100 g)
Total phenolics 1
Total carotene -0.970 1
ABTS 0.935* -0.822 1
DPPH 0.962* -0.867 0.996* 1
Reducing sugars -0.900 0.767 -0.995 -0.984 1
Vitamin C 0.891* -0.974 0.675 0.735 -0.605 1
Correlation coefficient statistically significant *pİ0.05
CONCLUSIONS Aminifard M.H., Aroiee H., Azizi V, Nemati H., Hawa

Z., Jaafar E., 2012. Effect of Humic Acid on
Antioxidant Activities and Fruit Quality of Hot
The differences between the variants Pepper (Capsicum annuum L.). Journal of Herbs,
determined by the Lignohumat biofertilizer Spices & Medicinal Plants, 18: 360-369.
were due both to the quality, the balance of the Becherescu A., Hoza Gh., Poșta G., Popa D., 2016.
nutrients contained in the composition and also Importance of crop substratum and foliar fertilizers on
to the treatment doses. Thus, the 200 g/ha yield and quality in hybrid cornichon cucumbers.
Journal of Horticulture, Forestry and Biotechnology,
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dose significantly influenced the antioxidant Investigaciones Agropecuarias 38: 1-6.
capacity compared to the untreated variant. Canellas L.P., Fábio L. Olivares, Natália O. Aguiar,
Davey L. Jones, Nebbioso A., Mazzei P., Piccolo A.,
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influence of biofertilization on a single variety, horticulture. Scientia Horticulturae, 196: 15-27.
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regarding the antioxidant properties and quality the surrounding environment and the alternative to the
attributes suggest that research can contribute chemical fertilizers. Ies- Envis New Sletter, 7 (3): 4-5.
Dinu M., Soare R., Bita G., 2017. The Effect Of Organic
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482
EFFICACY OF PLANT BASED BIOPRODUCTS AGAINST TOMATO

SEEDLINGS DAMPING-OFF DISEASE-SHORT OVERVIEW
Steliana RODINO, Marian BUTU, Alina BUTU
National Institute of Research and Development for Biological Sciences of Bucharest, 296 Splaiul
Independenţei, P.O. Box 17-16, 060031, Bucharest, Romania, Tel./Fax. +4021 220 0880
Abstract
Most European countries are investing in research to reduce reliance on pesticides and the risks associated with their
use. At this moment, farmers’ access to a wide range of pesticides is predicted to become limited due to legal
regulations regarding sustainable use of pesticides. Therefore, the modern farmers will have to incorporate innovative
pest and disease management approaches to reduce their dependency on pesticide use. This study presents an overview
about the recent results on the efficacy of natural products obtained from plant extracts to control tomato seedlings
damping off disease. It was already demonstrated that several plants contain secondary metabolites that are toxic to
plant pathogenic microorganisms infecting horticultural crops, especially on the early emergence of the disease. The
rich phenolic compound solutions possess antimicrobial effects and serve as plant defense mechanisms against plant
pathogens. For using these products with reproducible efficiency, it is important to compare their mode of action for
the optimization of the manufacturing process, the stabilization of these preparations, dates and rates of application.
Key words: plant bioproducts, tomato disease, plant extract.
INTRODUCTION causes emerging seedlings to collapse and once

infected, the seedlings rarely survive to produce
Plant diseases cause significant economical a vigorous plant. The period of growth between
loses in horticultural production every planting and maturity, is considered as critical,
cultivation season. Despite the continuous and special attention should be paid to protect
research on resistant cultivars, the crop losses sensitive seedlings. As a potentially useful
are still being observed. More often, the protection strategy, the use of plant products
intensive use of chemical pesticides resulted in such as essential oils and plant extracts against
inappropriate application of doses and has not several plant pathogens has already been
only created fungicide resistance and increased demonstrated. Tomato (Lycopersicon
soil contamination, but may also trigger esculentum Mill.) belongs to nightshade family
imbalance in the microbial community (On et Solanaceae. It is the second extensively grown
al., 2015) and a degradative effect on the ozone vegetable crop after potato due to its tangy
layer (Goudjal et al., 2014). Moreover, the fruit, taste and high nutritive value containing
access of farmers to a wide range of pesticides vitamins particularly vitamin A, C, β-carotene
is predicted to become limited due to legal and essential minerals (Chohan et al., 2017;
regulations regarding the implementation of Goudjal et al., 2014).
integrated pest management and sustainable use This study presents an overview about damping
of pesticides. As a result, most countries assign off tomato seedlings pathogens and the
important resources to search for solutions to reported efficacy of natural products obtained
reduce reliance on pesticides and the risks from plant extracts to control this disease.
associated with their use. The occurrence of
serious plant diseases can occur at the early CAUSAL PATHOGENS FOR DAMPING-
stage of seed germination and seedling OFF OF TOMATO SEEDLINGS
emergence. Damping off disease and rot of
crown and root of seedlings is caused by Almost all species of vegetables (tomatoes,
several different phytopathogens. This disease cucumber, eggplants, radishes, beans) can be
483
affected by damping off. The young leaves, emerged from the soil wards Table 1 (Kato et
roots and stems of newly emerged seedlings are al., 2013; Pagoch et al., 2015).
highly susceptible to fungal or fungus like The causal agents of damping off are soil borne
infections. pathogens that are also able to survive quite
When meeting proper conditions, damping-off well in plant debris, on contaminated tools, pots
pathogens may also cause the rot of root or and potting media. Their spores can be carried
crown rot in mature plants. Cool and cloudy by insects like fungus gnats, or can be
weather, excessive irrigation resulting in wet or transported in in irrigation pipes and drainage
compacted soil, poor ventilation and high water. Some pathogens can be introduced on
humidity due to overcrowding are just few of dirty hands, contaminated tools or by hose ends
the prerequisites for damping-off. that have been in contact with dirt and debris.
Damping off may occur before or after seed Rhizoctonia spp. and Fusarium spp., along with
germination. In pre-emergence, the seeds will the water mold Pythium spp. and Phytophthora
basically fail to germinate after sowing. They spp. are the most common pathogens
appear soft, dried or mushy, with necrotic responsible for damping off of many vegetables
radicle. In post-emergence damping-off, the seedlings, including tomato (Lycopersicon
seedlings will decay shortly after they have esculentum) (Chohan et al., 2017).
Table 1. Key features of fungi or fungal like organisms responsible for attacks
on tomato seeds and seedlings (adapted from Blancard, 2012)
Pathogen Symptoms Attack frequency Differentiating structures

Pythium spp. Soft wet brown rot on the Frequently Oospores, sporangia, and
(damping-off) stem base and roots. chlamydospores
Phytophthora spp Brown rot on the stem and Frequently Oospores, sporangia, and
P. erythroseptica, roots. chlamydospores
P. citricola,
P. capsici,
P. cryptogea,
P. nicotianae
(damping-off)
Sclerotium rolfsii Brown rot surrounding the Mainly in traditional White mycelium covering
(damping-off, stem and extending to the nurseries in tropical seedlings and soil.
stem base rot) roots level. areas. Spherical sclerotia (1-3 mm),
white to brown.
Rhizoctonia solani Reddish brown wounds on Quite frequently Compartmentalized mycelium,
(damping-off, hypocotyl and epicotyl before with a constriction at branches.
stem base rot) emergence.
After emergence, reddish
brown to black spots, on the
seedlings
The control and prevention of these pathogens PLANT PRODUCTS WITH

is mainly done by application of agrochemicals. ANTIFUNGAL ACTIVITY
However, the present movement towards
environmentally safe methods in sustainable In a study reported by Singh et al. (2012), four
agriculture calls for reducing their use. plant products obtained from leaf extracts were
Recent research directions are focusing on tested for their potential antifungal activity
developing environmentally friendly products against Pythium aphanidermatum, the causal
for the management of plant diseases. pathogen of damping-off of tomato seedling.
Natural plant products are considered a All products showed significant inhibitory
potential solution as they are important sources effect on the mycelial growth of the test
of new agrochemicals for the control of plant pathogen, as compared to chemical fungicide
diseases (Nashwa et al., 2012). Furthermore, control (Bavistin 0.2%). The leaf extract from
plant-based products are easily biodegradable. Eucalyptus globulus (92.78%) showed
maximum mycelial growth inhibition followed
484
by Azadirchta indica (72.91%) extract (Singh research, fifty five species of medicinal plants
et al., 2012). were employed for in vitro antifungal activity
The antifungal activities of plant products are testing against Rhizoctonia solani AG 2-1 and
attributed to different chemical compounds like Trichoderma harzianum. The aim was to
phenols, flavonoids, isoflavonoids, coumarins, improve the biocontrol efficacy of T.
pyrones, alkaloids etc. present in these plants harzianum. Cinnamomum loureirii stem bark
which effect the growth of pathogenic fungi. extract inhibited mycelial growth of R. solani
For example, the main components of AG 2-1 by 73.7%. The combination of T.
Eucalyptus sp. essential oil, determined by GC- harzianum and Cinnamomum loureirii stem
MS are β-Cymene (32.1%), Eucalyptol bark powder reduced the severity of radish
(36.59%), Cryptone (4.35%) and Spathulenol damping-off by 80.6%, suggesting that it can
(3.82%) (Rusu et al., 2014). be employed for the control of R. solani
In a recent study, Ocimum species essential oils development (Lee et al., 2011).
were evaluated for their in vitro antifungal Islam et al. (2012) evaluated the seed treatment
effect through poison food technique against with neem leaf (Azadirachta indica), garlic
Rhizoctonia solani and Choanephora clove (Allium sativum), allamanda leaf
cucurbitarum. Ocimum tenuiflorum, O. (Allamanda cathartica), ginger rhizome
gratissimum, and O. kilimandscharicum (Zingiber officinale), kalijira seed (Nigella
exhibited complete growth inhibition of both sativa), bel leaf (Aegle marmelos), turmeric
pathogens (24 and 48 h after treatment). rhizome (Curcuma longa), katamehedi leaf
Nevertheless, O. basilicum showed variable (Lawsonia alba) and onion bulb (Allium cepa)
levels of fungal growth inhibition (63.0%- against damping-off, seed germination and
100%). Moreover, the composition of the tested growth characters of tomato (Lycopersicon
essential oils was analyzed and compared by esculentum L.), eggplant (Solanum melongena
using capillary gas chromatography (GC/FID) L.) and chilli (Capsicum annum) seedlings. The
and GC-mass spectrometry (GC/MS). Phenyl most effective against seedling damping off
propanoids (upto 87.0%) and monoterpenoids was neem leaf extract followed by garlic clove
(upto 83.3%) were prevalent constituents and allamonda. The highest seed germination
distributed in the studied Ocimum taxa (Padalia of tomato seeds (86.67 %) was observed under
et al., 2014). neem leaf extract effect (Islam et al., 2012).
An essential oil of O. basilicum and Therefore, several papers were identified for
Cymbopogon citratus was also tested against presenting results in controlling a wide range of
three Phytophthora species under in vitro and tomato soil borne pathogens by different plant
greenhouse conditions compared to fungicides. products but still very few studies went further
However, C. citratus had the lowest EC50 to report the efficacy of botanicals against soil
values for in vitro inhibition of the mycelial borne pathogens in field level (Islam et al.,
growth of Phytophthora spp (between 31.5 and 2012). After reviewing the literature on
69.1). In greenhouse, C. citratus essential oil antimicrobial activity, it can be concluded that
reduced disease severity from 47.4% to 60.5% the vegetal material is representing an
compared to the untreated control (Amini et al., inexhaustible source of biologically active
2016). More recently, another Ocimum specie compounds. It was already demonstrated that
(namely Ocimum gratissimum) essential oil secondary metabolites of plant origin have
was tested for antifungal activity against practical application in the food industry and
several isolates of Fusarium spp., Rhizoctonia (Arsene et al., 2015) and agriculture, for
solani, and Macrophomina phaseolina. The sustainable crop disease management as well.
broth microdilution method revealed that F.
oxysporum f. sp. lycopersici and R. solani were CONCLUSIONS
the most sensitive strains (Mohr et al., 2017).
Rhizoctonia solani is very common soil fungus Due to international trend for environmentally
and is widespread in the world. It can be safe crop production, the modern farmers will
considered as a biomarker of so called ‘sick’ have to incorporate innovative pest and disease
soil (Blancard, 2012). In a more extensive management approaches to reduce their
485
dependency on pesticide use. Plant products isolated from native plants of Algerian Sahara,
have shown promising results for the control of Microbiological Research, 169 (1): 59-65.
Islam M.T., Faruq A.N., 2012. Effect of some medicinal
soil-borne pathogens. The antifungal and plant extracts on damping-off disease of winter
antimicrobial effects are due to synergistic vegetable, World Applied Sci. J., 17 (11): 1498-1503.
activity of constituents present in plant-based Kato M., Minamida K., Tojo M., Kokuryu T.,
products. Hamaguchi H., Shimada S., 2013. Association of
Nevertheless, for using these products with Pythium and Phytophthora with pre-emergence
seedling damping-off of soybean grown in a field
reproducible efficiency, it is important to converted from a paddy field in Japan. Plant Prod.
compare their mode of action for the Sci. 16: 95-104.
optimization of the manufacturing process, the Lee H.M., Khan Z., Kim S.G., Baek N., Kim Y.H, 2011.
stabilization of these preparations, dates and Evaluation of the biocontrol potential of some
rates of application. medicinal plant materials alone and in combination
with Trichoderma harzianum against Rhizoctonia
solani, Plant Pathol. J., 27 (1): 68-77.
ACKNOWLEDGEMENTS Mohr F.B., Lermen C., Gazim Z.C., Gonçalves J.E.,
Alberton O., 2017. Antifungal activity, yield, and
composition of Ocimum gratissimum essential oil.
This work was supported by a grant of the Genet Mol Res. 16 (1). Doi:10.4238/gmr16019542.
Romanian National Authority for Scientific Nashwa S.M.A., Abo-Elyousr K.A.M., 2012. Evaluation
Research and Innovation, CNCS/CCCDI - of various plant extracts against the early blight
UEFISCDI, project number PN-III-P2-2.1- disease of tomato plants under greenhouse and field
PED-2016-1544, contract number 14PED / conditions, Plant Protection Science 48 (2), 74-79.
On A., Wong F., Ko Q., Tweddell R.T., Antoun H., Avis
2017, within PNCDI III. T.J., 2015. Antifungal effect of compost tea
microorganism on tomato pathogens. Biol Control.
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486
DETERMINATION OF STRENGTH PROPERTIES

FOR MECHANICAL HARVEST OF PARSLEY
(PETROSELINUM CRISPUM)
Mehmet Emin GOKDUMAN, Deniz YILMAZ
Department of Agricultural Machinery and Technologies Engineering, Faculty of Agriculture,

Suleyman Demirel University, Isparta, Turkey
Corresponding author email*: [email protected]
Abstract
Parsley (Petroselinum crispum) is a vegetable from Umbelliferae family (or Apiaceae) that eats leaves as a salad.
Although parsley vegetable to produce small areas our country, it has started to make production in large and larger
areas in recent years. This study aimed to determine the strength of Parsley (Petroselinum crispum) specifications for
mechanical harvesting. For this purpose, properties as the maximum force, stress in the maximum force point, work at
maximum force point, shearing force, deformation at maximum force, bioyield force, and shearing stress of Parsley
(Petroselinum crispum) stalk, flower have determined. Average values for maximum force, stress and energy in
maximum force were determined as 4.535 N, 0.566 MPa and 0.015 J at stalk, respectively. The shearing force and
shearing stress were found to be as 1.170 N and 0.14 MPa, respectively. Average values for bioyield force were
determined to be 3.628 N. These features can be used in determining the design and operating conditions for the
mechanical harvester cutting blade.
Key words: Parsley (Petroselinum crispum), strenght properties, mechanical harvesting.
INTRODUCTION It has 23 000 (1000 ha) of farmland in Turkey.

3.4 percent of this area (809 000 ha) used for
Petroselinum crispum (Parsley) is a bright vegetable production.
green, biennial herb, which belongs to the Vegetable production has been increasing in
family Apiaceae (Figure 1). Native to the recent years.
central Mediterranean region (Southern Italy, According to 2016 data, the parsley production
Algeria and Tunisia) and naturalised elsewhere area is 49296 ha, the production volume is
in Europe, Africa and Asia. There are also 58160 tons in Turkey.
plenty of wild parsley in Spain, Greece, Depending on the variety and the season, 60-70
Morocco and Turkey. It is commonly used as a days after seeding, the plants are came to the
garnish in soups, salads, meats, vegetables and harvesting stage. Parsley harvest can be done in
sauces (Lis-Balchin, 2006). Traditionally, the 6-7 forms per a year.
leaf, seed and root are being used in herbal The vegetable mechanization is mostly
medicine (Simon and Overley, 1986). conducted by hand in Turkey. Mechanization is
needed due to the increase in production area.
The studies generally focused on chemical,
medicinal and culinary of parsley
(Petroselinum crispum) (Charles, 2004).
However, studies on strength properties of
parsley (Petroselinum crispum) are limited.
This study covers determination of maximum
force, bioyield force, shearing force, stress and
energy in maximum force, shearing
deformation and shearing stress of parsley
Figure 1. Parsley (Petroselinum crispum) plant (Petroselinum crispum) stalk, leaf.
487
MATERIALS AND METHODS The shearing forces on the load cell with
respect to knife penetration were recorded by
For this study, parsley (Petroselinum crispum) computer.
plants were harvested by hand from the parsley The shearing stress in N/mm2 was calculated
plant harvested from a greenhouse in the using the equation of Shahbazi et al. (2012):
Isparta province, Turkey. Fs max
Diameter and cross-sectional area of the
experimental samples were measured before A (1)
the shearing tests. Moisture content of the Where Fsmax is the maximum shearing force of
plants was determined at harvest time. the curve in N, and A is the area of the stalk at
Specimens were weighed and dried in an oven the deformation cross-section in mm2.
at 102°C for 24 h and then reweighed (ASABE,
2006). It was provided concise but complete
information about the materials and the The parsley plants were attached to the
analytical and statistical procedures used. apparatus from its stalks (Figure 4). The
A universal testing machine (LF Plus, UK) shearing tests were conducted with 0.8 mm s˗1
with a 500 N load cell and a computer-aided knife speed progress (Simonton, 1992).
cutting and picking apparatus (Figures 2 and 3)
was used to measure the strength properties of
the parsley (Petroselinum crispum) plant. Knife
material was hardened iron. All the tests were
carried out at a speed 0.8 mm s-1, and data were
recorded at 10 Hz. All data were analyzed by
nexygen software program.
Figure 4. Measuring the cutting of parsley

(Petroselinum crispum) plant
Picking force can be defined as the force
Figure 2. Cutting system required to separate leaf stalks from ovary point
(picking force of leafs). The load cell of the
machine was then pulled upward to determine
the picking force of the parsley (Petroselinum
crispum) leaf (Figure 5).
Maximum force, bioyield force, shearing force,
stress and energy in maximum force, shearing
stress and shearing deformation were calculated
from the force-deformation curves at the
inflection point as defined by ASAE Standard
(1985). S368.1 (ASAE Standards, 1985) was
obtained from all curves (Figure 6).
The energy of shearing was determined as the
area under these curves (Chen et al., 2004;
Srivastava, 2006).
Figure 3. Picking system
488
Moisture content of the parsley plants was

determined as 85.6 % at harvest time and all
tests were conducted at harvest moisture. The
strength measurements of rocket (parsley)
stalks are given in Table 1.
The maximum force was observed as 4.535 N
at parsley stalk. The bioyield force of 3.628 N
was observed at stalk.
Shearing force is one of the most important
plant characteristics affecting plant harvesting.
If the weight of the plant is known, the shearing
force and the shearing height can be used to
determine the speed of the blade to be used in
Figure 5. Measuring the picking force of parsley harvesting (Igathinathane et al., 2010;
(Petroselinum crispum) leaf stalk
Taghijarah et al., 2011).
The maximum shearing force was observed as
1.170 N at stalk. The stress value in maximum
force (0.566 MPa) was observed at stalk.
The energy at maximum force was found to be
as 0.015 J.
Deformation has an important place among the
strength characteristics of the plant. The
maximum shearing deformation (24.323 mm)
was observed at stalk. The average cross-
sectional area of parsley (Petroselinum
crispum) was determined as 7.884 mm2 at
Note. Labels on the graph indicate the following points:
x - bioyield force, y - maximum force, z - shearing force (Liu, 2012)
harvest moisture (85.6%). The strength
measurements of parsley (Petroselinum
Figure 6. Typical force-deformation curve of parsley
(Petroselinum crispum) plant stalk during shearing
crispum) leaf are given in Table 2.
loading
Table 1. Average strength properties of parsley (Petroselinum crispum) stalk
Stress in Energy in
Maximum Bioyield Shearing Shearing Shearing
maximum maximum Area
force force force stress deformation
force force (mm 2)
(MPa) (mm)
(N) (N) (N) (MPa) (J)
Stalk 4.535 3.628 1.170 0.566 0.015 0.14 24.323 7.884
Standard Deviation 2.994 2.393 1.406 0.176 0.018 0.09 3.465 3.575
Table 2. Average strength properties of parsley (Petroselinum crispum) leaf
Stress in Energy in
Maximum Bioyield Shearing Shearing Shearing
maximum maximum
force force force stress deformation
force force
(MPa) (mm)
(N) (N) (N) (MPa) (J)
(Leaf) Flower 1.773 1.418 0.904 0.125 0.006 0.03 6.308
Standard Deviation 0.823 0.658 0.002 0.009 0.001 0.01 1.063
489
The maximum force required to separate leaf REFERENCES
from stalk was determined as 1.773 N. As a
function of the maximum force the bioyield ASABE Standards, 2006. Moisture measurement e
force was found to be 1.418 N. Lower shearing Forages. St. Joseph, MI: American Society of
Agricultural and Biological Engineers (ASABE).
forces required for mechanical harvesting leads S358.2
to savings in power and energy usage. Leaf ASAE Standards, 1985. Compression test of food
shearing force of parsley observed 0.904 N is materials of convex shape. St. Joseph, Mich.:
lower than stalk shearing force. The maximum American Society of Agriculture Engineering, S368.1
stress in maximum force value (0.125 MPa) Charles D.J., 2004. Parsley. In: Peter KV (ed), Handbook
of Herbs and Spices, pp. 230-242. Cambridge:
was observed at leaf. The energy at maximum Woodhead Publishing Limited, CRC Press.
force was found to be as 0.006 J. The stress Chen Y., Gratton J.L., Liu J., 2004. Power requirements
shearing value was observed as 0.03 MPa. of hemp cutting and conditioning. Biosystems
The average shearing deformation value of Engineering, 87 (4), 417-424.
parsley leaf was found as 6.308 mm. Igathinathane C., Womac A.R., Sokhansanj S., 2010.
Corn stalk orientation effect on mechanical cutting.
Biosystems engineering, 107 (2), 97-106.
CONCLUSIONS Lis-Balchin M., 2006. Aromatherapy science: a guide for
healthcare professionals. Pharmaceutical press.
This study was carried out to determine the Shahbazi F., Nazari Galedar M., 2012. Bending and
strength properties of parsley (Petroselinum shearing properties of safflower stalk. Journal of
Agricultural Science and Technology, 14 (4), 743-
crispum) at leaf and stalk sections in the 754.
harvest moisture. Properties as the maximum Simon, J.E., Overley M.L., 1986. A comparative
force, bioyield force, shearing force, stress in evaluation of parsley cultivars. The Herb, spice and
maximum force, energy in maximum force, medicinal plant digest (USA).
shearing stress, shearing deformation of parsley Simonton W., 1992. Physical properties of zonal
geranium cuttings. Trans. ASAE, 35(6): 1899-1904.
(Petroselinum crispum) leaf and stalk have Srivastava A.K., Goering C.E., Rohrbach R.P.,
determined at moisture content of 85.6%. Buckmaster D.R., 2006. Engineering principles of
The strength parameters measured at root agricultural machines (2nd ed.). American Society of
section higher than that of the stalk and leaf Agricultural and Biological Engineers, St. Joseph,
sections. The lowest values were found at USA, 185.
Taghijarah H., Ahmadi H., Ghahderijani M., Tavakoli
parsley (Petroselinum crispum) stalk. The M., 2011. Shearing characteristics of sugar cane
strength parameters of stalk section should be (Saccharum officinarum L.) stalks as a function of the
considered for mechanical harvesting of rocket rate of the applied force. Australian Journal of Crop
plant to provide data for the design machines Science, 5 (6), 630.
for mechanized applications.
490
WHICH IS CRUCIAL FOR HETEROSIS? TRAITS,

GENETIC OR CHARACTERISTIC DIVERSITY:
PUNGENCY PARADIGM
Bekir Bülent ARPACI1, Cihan FIRAT2, Mehmet KOÇ1, Faika YARALI KARAKAN1
1
Kilis 7 Aralık University, Faculty of Agriculture, Department of Horticulture,
Mehmet Sanlı Mah. Doğan Güreş Paşa Bul. No: 134 Kilis, Turkey
2
East Mediterranean Transitional Zone Agricultural Research Institute,
Hacimustafa Mah. Gazi Mustafa Kemal Bulvari No: 122, Kahramanmaras, Turkey
Abstract
Commercially plant breeding and crop improvement is possible to know heterotic behavior of plant species and
heredity of the traits. It is commonly accepted that heterosis has been positively correlated with distance between traits
of the parents. Two pepper variety belongs different Capsicum species, non-pungent Santos Flame (Capsicum annuum
L.) and Biquinho (Capsicum chinense Jacq.) which had very low capsaicinoid content were crossed to estimate mid
parent heterosis, heterobeltiosis and standard heterosis. In this study extraordinary heterosis rates were calculated F1
progeny of two different Capsicum species close to pungency trait but definitely divergent genetically. The capsaicin
content of Santos Flame, Biquinho and F1 progeny were 0 mg kg-1, 2.1 mg kg-1 and 217.4 mg kg-1, respectively. The
highest mid parent heterosis was estimated from capsaicin content with 20605%. Similarly uncommonly heterobeltiosis
and standard heterosis rate was estimated for capsaicin content with 10252% and 237%, respectively. Correlatively
extreme heterosis rates were calculated for SHU pungency level, dihydrocapsaicin and total capsaicinoids content.
Key words: pungency, heterosis, heterobeltiosis, pepper.
INTRODUCTION and homodihydrocapsaicin (Greenleaf, 1986;

Collins and Bosland, 1994).
The importance of heterosis in agriculture is to The capsaicinoid content of the pepper
improve high yielded varieties. Heterosis is not cultivars varies greatly and related with
a trait expressed by specific genes or alleles maturity. A subjective assessment Scoville
therefore distinction is important for heterosis Heat Unit (SHU) based on panel evaluation
(Lippman and Zamir, 2007). Genetic diversity also is used to determine pungency of peppers.
could be as great value as combining ability or ASTA (American Spice Trade Association)
greater than it (Hayes and Immer, 1942). Allard prefers Scoville Heat Unit determining of
(1999) indicated that single-cross hybrids pungency (Scoville, 1912; ASTA, 1985).
diverged maximally in allelic frequencies With this study two different Capsicum species
produce most heterozygoty in most loci in F1 distinct from genetically but close to pungency
hybrids. trait were crossed and inheritance of pungency
The most important quality characteristics in component calculated.
pepper are color, pungency, dry/fresh yield
ratio and flesh thickness (Abak, 1995). MATERIALS AND METHODS
Capsaicinoids are colorless, odorless and
flavorless substances only found in pepper. Two pepper cultivars ‘Santos Flame’,
They have burning feature and no nutrition PanAmerican Seed Illinois USA (Capsicum
value. Cause of heat feeling substances are annuum L.), as female, and ‘Biquinho’ (C.
capsaicinoids consist of the most important chinense Jacq.), as male, were crossed and
seven different substances structurally similar capsaicinoid contents of parents, F1 48 F2,
pungent compounds including capsaicin, progenies and check variety Jalapeño were
dihydrocapsaicin, nordihydrocapsaicin, measured by high pressure liquid
norcapsaicin, homocapsacin, nornorcapsaicin chromatography.
491
Extraction of capsaicin and mean of retention time was 5.56 min. for
dihydrocapsaicin capsaicin and 7.47 min. for dihydrocapsaicin.
The whole fruits dried at 50°C for 2 days and Cultivar Biquinho which had a little amount
ground to fine powder. One gram of dried and (2.10 mg kg-1) capsaicin content crossed non-
ground fruit stirred in 10 ml sodium acetate pungent Santos Flame and hybrid exhibited 217
saturated ethanol during 3 hours in 60°C. The mg kg-1 capsaicin. An extraordinary heterosis
mixture was filtrated and analyzed. over mid parent, better parent and standard
check (Jalapeño 64.60 mg kg-1) was observed
Determination of capsaicin and in F1 generation for capsaicinoid content and
dihydrocapsaicin contents pungency level from interspecific crosses of
HPLC: was (Model LC-20 Shimadzu, Kyoto, Capsicum. The magnitudes of heterosis, better
Japan) consisting of a high-pressure pumps, parent and standard check for capsaicin were
column (C18 100-5 250 × 4.6 mm), oven 20605, 10252 and 237%, respectively (Table
(35°C) and UV-VIS detector (at 280 nm). 1.)
Isocratic mode (methanol 48.4%, water 30.2% Mid parent heterosis and heterobeltiosis was
dioxan 13.3%, acetonitrile 7.9% and 3877 and 1889%, respectively while the
perchloric acid 0.2% [% 2]) was performed at economic heterosis was 228% over standard
1.5 ml min−1 of flow rate with 20 μL sample check Jalapeño cultivar on account of
volume (ASTA, 1985). dihydrocapsaicin.
Standards of capsaicin and dihydrocapsaicin The percentage of mid parent heterosis related
were purchased from Sigma-Aldrich Co. sum of the capsaicin and dihydrocapsaicin
Pungency values were converted to SHU contents was 7855%. Heterobeltiosis and
(Scoville Heat Unit) multiplying total standard heterosis was 3877 and 233%, respec-
capsaicinoids by 15 (Mathur et al., 2002). tively. Correlatively pungency level had simi-
The plants of parents and progenies were larities with sum of the capsaicin and dihydro-
grown in field conditions during the 2015 capsaicin concerning mid parent, better parent
pepper growing season in EMTZARI (East and economic heterosis were found same.
Mediterranean Transitional Zone Agricultural The capsaicin contents ranged between 0.00 -
Research of Institute Kahramanmaraş, Turkey). 1279 mg kg-1 and the dihydrocapsaicin contents
ranged from 0.00 - 912 mg kg-1. The Accession
Calculating heterosis 97 had the highest capsaicin content with 1279
Mid parent, better parent also known as mg kg-1 while the highest dihydrocapsaicin was
heterobeltiosis and standard also called as obtained from Accession 103 with 912 mg kg-1.
economic heterosis (Sharma et al., 2013) Ten accessions from F2 progeny of Santos
calculated by using formula. Flame × Biquinho had no capsaicin content.
F1‒MP Dihydrocapsaicin was not detected from 11
Mid Parent = × 100 accessions of F2 progeny of Santos Flame ×
MP
Biquinho. Despite low level capsaicin content
F1‒BP than Acc. 97, Acc. 103 was the most pungent
Better Parent= × 100
BP genotype with 30793 SHU because of its high
F1‒Check (Jalapeño) amount of dihydrocapsaicin. Segregation of all
Standard= × 100 of the pungency components transgressively in
Check (Jalapeño)
F2 progeny demonstrated that pungency was
Where: F1 was mean of F1; MP was mean of expressed dominantly (Figures 2 and 3).
two parents; BP was mean of better parents and Biquinho exhibited 140 SHU pungency level
Jalapeño was used for control variety. while it was 0 in Santos Flame. The
interspecific hybrid of these two cultivars
RESULTS AND DISCUSSIONS produced 349.61 mg kg-1 sum of capsaicin and
dihydrocapsaicin content and exposed 5594
The chromatogram peaks for capsaicin and SHU pungency level. Pungency mean of
dihydrocapsaicin of parents, F1, Acc. 97, Acc. parents was 70 SHU and the check variety
103 and 270) were exhibited in Figure 1. The Jalapeño had 1678 SHU level.
492
Table 1. Capsaicinoid content of hybrid, parental and check varieties and heterosis percentage of pungency components
Capsaicin Dihydrocapsaicin Capsaicin + Dihydrocapsaicin
Pungency (SHU)
(mg kg-1) (mg kg-1) (mg kg-1)
Biquinho 2.10 6.65 8.79 140
Santos Flame 0.00 0.00 0.00 0
Mean 1.05 3.33 4.40 70
F1 217.40 132.25 349.61 5594
Jalapeño 64.60 40.26 104.85 1678
Heterosis (%)
Mid Parent 20605 3877 7855 7855
Better Parent 10252 1889 3877 3877
Standard 237 228 233 233
Figure 1. Capsaicin and dihydrocapsaicin HPLC chromatogram of Biquinho,

Santos Flame and their F1 and 103, 97,270 lines from F2 progenies
2500
2000
1500
1000
500
0
50
39
81
61
71
66
57
28
97
29
99
87
84
68
95
30
48
2
Biquinho
270
180
240
288
217
117
148
205
140
256a
120
260
8
219
103
6
109
222
285
256
136
278
253
248
117a
273a
39a
123
228
273
F1
Jalapeño
Santos Flame
Capsaicin Dihydrocapsaicin Caps+DhCaps
Figure 2. Capsaicin, dihydrocapsaicin (mg kg-1) content of Biquinho, Santos Flame and their F1 and F2 progenies
35,000
30,000
25,000
20,000
15,000
5,594
10,000
15,449
18,171
22,923
30,793
27,036
17,962
14,308
13,718
11,679
11,557
2,164
3,958
4,054
4,057
4,415
4,671
5,493
6,082
9,426
7,327
5,203
4,825
4,216
3,846
2,686
1,466
1,678
2,113
206
220
297
607
588
232
228
209
140
5,000
33
77
75
57
43
0
0
0
0
0
0
0
0
0
50
39
81
61
71
66
57
28
97
29
99
87
84
68
95
30
48
2
Biquinho
256a
117a
273a
F1
Jalapeño
270
180
240
288
217
117
148
205
120
260
140
219
103
109
222
285
256
136
278
253
248
39a
123
228
273
Santos Flame
Pungency (SHU)
Figure 3. Pungency level (SHU) of Biquinho, Santos Flame and their F1 and F2 progenies
493
The pungency components in pepper were first quality traits such as pungency could be more
extracted by Thresh (1846) and named as heterotic compared yield and yield component.
capsaicin. Substances of pungency in peppers
are highly influenced by environmental ACKNOWLEDGEMENTS
conditions such as high temperatures and water
stress (Lindsey and Bosland, 1996). Ahmed et The pungency measurements were supported
al. (1982) emphasized that the pungency is a by Advanced Technology Application and
dominant character and additive gene effect Research Center of Kilis 7 Aralık University.
was very important in their inheritance.
According to our results transgressive REFERENCES
segregation of pungency indicated capsaicin,
dihydrocapsaicin content and pungency was Abak K., Pakyurek Y., 1995. Processes needed to take
inherited dominantly. into consideration at the red pepper farming in K.
Maras. Sutcu Imam University Publications, 5-6.
Biquinho (Capsicum chinense Jacq.) is known Ahmed N., Singh J., Bajaj K.L., 1984. Genetics of
as non-pungent sweet pepper cultivar (Alves et Capsaicin content in chilli (Capsicum annuum L.).
al., 2014; Sganzerla et al., 2014). Nevertheless Journal of Research Punjab Agricultural University.
de Aguiar et al. extracted (2014) low Allard R.W., 1999. Principles of plant breeding. John
concentration of capsaicinoids in Biquinho Wiley & Sons.
Alves V.C.C., Pinto N.O., Penha M.F.A., Gomes B.L.,
pepper. We found a low concentration of Reifschneider F.J., Garruti D.S., 2014. Aroma-Active
capsaicin and dihydrocapsaicin in Biquinho. Compounds of Capsicum chinense var. Biquinho. In
These two compounds were not detected from Flavour Science, 567-571.
Santos Flame. The main objective of the American Spice Trade Association, 1985. Official
research was to manifest that genotypes close analytical methods of the American Spice Trade
Association, 3rd ed. Amer. Spice Trade Assn.,
to concerned trait could be exhibited heterotic Englewood Cliffs, N.J.
behavior if they were distinct from genetically. De Aguiar A.C., Dos Santos P., Coutinho J.P., Barbero
Prasad and Singh (1986) indicated that G.F., Godoy H.T., Martínez J., 2014. Supercritical
extremely divergent parents create high fluid extraction and low pressure extraction of
magnitude of heterosis harmonizing by alleles. Biquinho pepper (Capsicum chinense). LWT-Food
Science and Technology, 59 (2), 1239-1246.
Genetic diversity is important related with Hayes H.K., Immer F.R., 1942. Methods Of Plant
heterosis and inbred lines from different origin Breeding. Mcgraw-Hill Book Company, Inc.; New
show greater heterosis (Hayes and Immers, York; London.
1942). Pearson (1983) reported that Peter and Lindsey K., Bosland P.W., 1995. A field study of
Singh (1976) declared 494% heterosis related environmental interaction on pungency. Capsicum
Eggplant Newsl, 14, 36-38.
with yield in eggplant and Shifriss and Rylski Lippman Z.B., Zamir D., 2007. Heterosis: revisiting the
(1973) notified 195% for exported quality in magic. Trends in genetics, 23 (2), 60-66.
bell pepper. Tu et al. (2007) find 672.7% better Pearson O.H., 1983. Heterosis in vegetable crops. In
parent heterosis for kernel yield in rice. Heterosis, Springer, Berlin, Heidelberg, 138-188
However there is no extraordinary heterosis Prasad S.K., Singh T.P., 1986. Heterosis in relation to
genetic divergence in maize (Zea mays L.).
and heterobeltiosis percentage as we found in Euphytica, 35 (3), 919-924.
our experiment for the literally we know. Scoville W.L., 1912. Note on capsicums. Journal of
Pharmaceutical Sciences, 1 (5), 453-454.
CONCLUSIONS Sganzerla M., Coutinho J.P., De Melo A.M.T., Godoy
H.T., 2014. Fast method for capsaicinoids analysis
from Capsicum chinense fruits. Food research
The extreme heterosis and heterobeltiosis international, 64, 718-725.
percentages were observed from hybrid of two Singh R.K., Chaudhary B.D., 1985. Line x Tester
species Capsicum chinense Jacq. and Capsicum analysis. Biometrical Methods in Quantitative
annuum L. distinct from genetically but closed Genetic Analysis, Ed, 3, 215-223.
to pungency trait and its component. Therefore Thresh L.T., 1846. Isolation of capsaicin. Pharm J, 6,
941.
genetic diversity was found important related Tu S., Luan L., Liu Y., Long W., Kong F., He T., Yu M.,
with heterosis. However it is believed that 2007. Production and heterosis analysis of rice
autotetraploid hybrids. Crop science, 47 (6), 2356-
2363.
494
THE EFFICACY OF DIFFERENT TREATMENTS FOR PATHOGENS

CONTROL ON THE EGGPLANT CROPS IN THE FIELD
Mihaela Alina BUZATU1, Marcel COSTACHE2, Dorel HOZA3,

Gabriela ȘOVĂREL2, Stelica CRISTEA3
1
Engineering and Management of Vegetal and Animal Resources Doctoral School,
UASMV Bucharest, 59 Mărăşti Blvd, District 1, Bucharest, Romania
2
Research and Development Institute for Vegetable and Flower Growing Vidra, No. 22,
Calea București Street, Vidra, Ilfov, Romania
3
In the field, eggplants are frequently attacked by Alternaria porri f. sp. solani, Botrytis cinerea and Phytophthora
parasitica pathogens, which diminishes yield in quantitative terms and depreciates qualitatively. For the control of
these pathogens three variants of fungicide treatments with different active ingredients were tested: chlorotalonil 500
g/l, pyraclostrobin 5% + metiram 55%, metiram 80%, iprovalicarb 8.4% + Cu oxychloride 40.6%, azoxystrobin 200 g/l
+ diphenoconazole 125 g/l. The best efficacy was obtained at variant 2 with the following schedule of treatments:
treatment 1 - pyraclostrobin 5% + metiram 55% - 0.2%; treatment 2 - iprovalicarb 8.4% + Cu oxychloride 40.6% -
0.2%; treatment 3 - pyraclostrobin 5% + metiram 55% - 0.2%; treatment 4 - iprovalicarb 8.4% + Cu oxychloride
40.6% - 0.2%; treatment 5 - azoxystrobin 200 g/l + diphenoconazole 125 g/l - 0.1%; treatment 6 - pyraclostrobin 5% +
metiram 55% - 0.2%. In this variant, the average efficacy was 85.5% and the yield 39.7 t/ha (123.17%) compared to
31.8 t/ha in the untreated control variant.
Key words: Alternaria porri f. sp. solani, Botrytis cinerea, pathogens control, Phytophthora parasitica, Solanum
melongena.
INTRODUCTION 2008; Radu et al., 2011; Pochon et al., 2012;

Mardare et al., 2014; Pană et al., 2014; Cristea
Although eggplants (Solanum melongena L.) et al., 2015; Manole and Cristea, 2015; Gruia et
are characterized by their adaptation to varied al., 2016; Dudoiu et al., 2016). The distribution
pedoclimatic conditions, crop yield is restricted of Alternaria spp. on various plant species
by the large number of pests whose attack (Berca et al., 2015) was also studied in the
affects the productive biological potential, literature.
causing the decrease of quantity and quality of Under laboratory conditions comparative
fruits (Tomescu et al., 1992; Costache and measurements were made on enzymantic
Roman, 2007). In the field eggplants are content of the leaves of Momordica charantia
frequently attacked by the pathogens Alternaria (both healthy and infected with pathogenic
porri f. sp. solani, Botrytis cinerea (Buzatu et. fungi Aspergillus niger and Alternaria sp.).
al., 2017), Phytophthora parasitica and soil Amylases, proteases and lipases content of the
borne fungal pathogen including Verticillium biological material were determined.(Cozea
dahliae and Fusarium oxysporum f. sp. and Cristea, 2011). In our country, this disease
melongenae (Buzatu et. al., 2017). Alternaria was first reported in 1958 by Tr. Săvulescu
porri f. sp. solani occur frequently in eggplant (Docea, 2012). Botrytis cinerea occurs
cultures, especially in years with high average frequently in crops of eggplants in protected
temperatures and heavy rainfall, producing areas, but also in the fields in years of abundant
dark brown spots on leaves, stems and flowers precipitation or if the culture is irrigated by
that coalesce and form large necrotic areas sprinkler irigation.
(Leite, 1997; Oliveira et al., 2004; Cristea, Under favorable environmental conditions to
2005). Alternaria spp. has also been identified the onset and evolution of the attack, the losses
on seeds of various plant species (Cristea et al., can reach up to 15-20%.
495
Symptoms present in flowers range from MATERIALS AND METHODS
browing to fall off. On the fruit, the symptoms
begin at the base of the calyx through a small The experiments were carried out in 2017,
round lesion that develops into a soft rot. under field conditions, using the Luiza eggplant
The fruit is covered by a mass of gray spores, variety. The culture was planting on May 12,
characteristic of the disease (Compendium of 2017, with a density of 28,000 plants/ha. Four
Strawberry Diseases, 1998). variants of treatments (Table 1) were used for
Phytophthora parasitica appears more the control of pathogens Alternaria porri f. sp.
frequently on well - developed fruits at the base solani, Botrytis cinerea and Phytophthora
of plants that come in contact or are near the parasitica. Technological control variants have
surface of the soil. The tissues of the attacked been establisheed according to the sequence of
fruit is rotting. pathogens in culture. During the growing season
Under high temperature and humidity were applied 6 foliar treatments, at intervals of
conditions, the rot is expanding and 12-16 days, in correlation with the climatic
progressing affecting the whole pulp (Docea, factors. Observations were made on the
2012). frequency (AF%) and the intensity (I%) of
Based on the results obtained in the world and pathogen attack and the degree of attack
presented in the literature (Dillard et al., 1996; (DA%) and efficacy (E%) were calculated. The
Anthony et al., 1998), the work aims to degree of attack was calculated using the
highlight the possibilities to monitor the formula (F% × I%)/100 and the efficacy with
complex of pathogens in the eggplant crops by the formula (untreated DA% ‒ treated DA%) ×
using different treatment schedule. 100/untreated DA.
Table 1. Variants to pathogens control on the eggplant crops in the field (Vidra, 2017)
22.06.2017 06.07.2017 18.07.2017 04.08.2017 22.08.2017 06.09.2017
Variant
Treatment 1 Treatment 2 Treatment 3 Treatment 4 Treatment 5 Treatment 6
I. chlorotalonil iprovalicarb 8.4% chlorotalonil iprovalicarb 8.4% azoxystrobin chlorotalonil
500 g/l - 0.2% + Cu oxychloride 500 g/l - 0.2% + Cu oxychloride 200 g/l + 500 g/l - 0.2%
40.6% - 0.2% 40.6% - 0.2% diphenoconazole
125 g/l - 0.1%
II. pyraclostrobin iprovalicarb 8.4% pyraclostrobin iprovalicarb 8.4% azoxystrobin pyraclostrobin
5% + metiram + Cu oxychloride 5% + metiram + Cu oxychloride 200 g/l + 5% + metiram
55% - 0.2% 40.6% - 0.2% 55% - 0.2% 40.6% - 0.2% diphenoconazole 55% - 0.2%
125 g/l - 0.1%
III. metiram 80 % - iprovalicarb 8.4% metiram 80 % - iprovalicarb 8.4% azoxystrobin metiram 80% -
0.2% + Cu oxychloride 0.2% + Cu oxychloride 200 g/l + 0.2%
40.6% - 0.2% 40.6% - 0.2% diphenoconazole
125 g/l - 0.1%
IV. Untreated - - - - -
control
RESULTS AND DISCUSSIONS been studied in the literature (Mardare et al.,

2015).
The first pathogen in culture was Botrytis The attack of the 3 pathogens had a slow
cinerea, followed by Alternaria porri f. sp. evolution due to the low relative atmospheric
solani and Phytophthora parasitica. The attack humidity in the period July - August (52.4 -
of the pathogens Botrytis cinerea and 74.7%, mean = 61.2%; Table 2).
Alternaria porri f. sp. solani began in the third Under these conditions, the average efficacy of
decade of July (21.07. and 24.07., respectively) treatment variants was between 79.7% (V III)
and the Phytophthora parasitica in the first and 85.5% (V II, Table 3).
decade of August (7.08.). Abiotic factors The yields of field eggplants were 38.7 t/ha (V
(temperature, atmospheric humidity and light) III), 39.7 t/ha (V II) and the untreated control
are important factors in the evolution of variant was 31.8 t/ha (Table 4).
pathogen attack. The highest yields were obtained in variants II
The influence of these abiotic factors on the (39.7 t/ha - 124.8%) and I (39.2 t / ha - 123.1%.
growth and development of Alternaria spp. has
496
Table 2. The occurrence and evolution of the pathogens attack on the
eggplants from the field in correlation with climatic factors (Vidra, 2017)
Date Degree of attack/frequency of attack (%)/month, decade
The pathogens/ of the May June July August September
climatic factors attack III I II III I II III I II III I II III
Alternaria solani 24.07 0 0 0 0 0 0 1,1 2.3 4.2 5.8 6.7 8.8 10.9
Botrytis cinerea 21.07 0 0 0 0 0 0 1.4 1.9 2.5 3.4 3.9 5.8 8.1
Phytophthora 7.08 0 0 0 0 0 0 0 0.8 1.5 2.3 3.2 4.4 5.9
parasitica
Minimum - 12.4 15.0 14.0 16.3 15.9 15.9 17.5 21.1 18.3 15.1 13.7 15.3 10.5
temperature (ºC)
Maximum - 21.4 27.0 26.0 31.9 28.4 28.9 31.0 36.6 31.8 28.7 28.8 29.4 19.2
temperature (ºC)
Average - 16.5 20.1 19.8 23.8 22.0 22.0 24.0 30.8 25.6 21.3 20.8 21.5 14.2
temperature (ºC)
Minimum relative - 57.0 47.5 43.8 36.4 46.0 37.8 35.7 26.9 27.4 30.2 29.3 30.8 41.7
humidity (%)
Maximum relative - 79.6 77.8 74.4 82.5 74.7 63.6 63.3 52.4 57.0 56.4 72.3 61.7 64.6
humidity (%)
Average relative - 66.9 59.5 56.8 56.4 57.9 47.6 46.8 36.7 38.7 41.7 46.9 44.8 50.7
humidity (%)
Precipitation (mm) - 7.5 20.0 22.5 1.0 84.0 8.5 6.5 0 0 45.0 30.0 1.0 2.0
Table 3. Efficacy of some schedules of trearments to control pathogens on the eggplant culture (Vidra, 2017)
Degree of attack / frequency of attack (%) and efficacy (%)
Average
Phytophthora
Variant Alternaria solani Botrytis cinerea of
E (%) E (%) parasitica E (%)
(DA%) (AF%) efficacy
(AF%)
(%)
I 2.2 79.8 1.3 83.9 1.0 83.0 82.2
II 1.9 82.6 1.0 87.6 0.8 86.4 85.5
III 2.4 78.0 1.5 81.5 1.2 79.7 79.7
IV
(Untreated 10.9 - 8.1 - 5.9 - -
control)
Table 4. The yield of eggplant in correlation with the experimental treatments (Vidra, 2017)
Variant Yield
% as compared to the Difference Significance
kg/sqm
untreated control (kg/sqm)
I 3.920 123.1 + 0.74 ***
II 3.972 124.8 + 0.79 ***
III 3.872 121.6 + 0.69 **
IV
3.182 100.0 0
(untreated control)
LSD 5% - 0.359 kg/ sqm, LSD 1% - 0.504 kg/sqm, LSD 0.1% - 0.712 kg/sqm
Analyzing the obtained results, compared to the In the climatic conditions of 2017, in Vidra,
untreated control variant, it was found that the Ilfov region, these pathogens decreased the
best results were obtained in the variants I and yield with 22-25%.
II, with very significant yield differences The best efficacy was obtained at variant II
compared to the untreated control variant. with the following schedule of trearments:
treatment 1 - pyraclostrobin 5% + metiram
CONCLUSIONS 55% - 0.2%; treatment 2 - iprovalicarb 8.4% +
Cu oxychloride 40.6 % - 0.2%; treatment 3 -
Alternaria porri f.sp. solani, Botrytis cinerea pyraclostrobin 5% + metiram 55% - 0.2%;
and Phytophthora parasitica are more frequent treatment 4 - iprovalicarb 8.4% + Cu
occured pathogens that attack the foliage and oxychloride 40.6% - 0.2%; treatment 5 -
fruits of the eggplants. azoxystrobin 200 g/l + diphenoconazole 125 g/l
- 0.1% and treatment 6 - pyraclostrobin 5% +
497
metiram 55% - 0.2%. In this variant the mean Dillard H.R., Johnston S.A., Cobb A.C., Hamilton G.H.,
efficacy was 85.5%. The highest yields were 1997. An assessment of fungicide benefits for the
control of fungal diseases of processing tomatoes in
obtained in variants II (39.7 t/ha - 124.8%) and New York and New Jersey. Plant Dis. 81 (6): 677-
I (39.2 t/ha - 123.1%). 681.
Dudoiu R., Cristea S., Lupu C., Popa D., Oprea M.,
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498
PATHOGENS WITH ECONOMIC IMPORTANCE FOR TOMATO CROPS

GROWING IN THE FIELD AND THEIR CONTROL
Iuliana Mândru1, Marcel Costache2, Dorel Hoza3, Stelica Cristea3
1
Engineering and Management of Vegetal and Animal Resources Doctoral School,
UASMV Bucharest, 59 Mărăşti Blvd, District 1, Bucharest, Romania
2
Research and Development Institute for Vegetable and Flower Growing Vidra, No. 22,
Calea București Street, Vidra, Ilfov, Romania
3
Faculty of Horticulture, 59 Mărăşti Blvd, District 1, Bucharest, Romania
Abstract
During 2017, at the RDIFG Vidra, was organized a bifactorial experience, placed on the subdivision parcels method,
with 12 variants and 4 repetitions, using Pontica 102 tomato variety. During the growing period on tomatoes have been
identified following pathogens on foliage: Pseudomonas syringae pv. tomato, Xanthomonas campestris pv. vesicatoria,
Alternaria porri f. sp. solani, Fulvia fulva and Phytophthora infestans. Their attack has influenced production in terms
of quantity and quality. In order to reduce yield losses, different treatment variants were tested using the following
fungicide-bactericids: copper hydroxide 50% (Copper Max 50 WP 0.25%), chlorothalonil 500 g/l (Bravo 500 SC
0.2%), azoxystrobin 200 g/l - difenoconazole 125 g/l (Ortiva Top 0.1%), iprovalicarb 8.4% + Cu of oxychloride 40%
(Melody Compact 49 WG 0.2%), difenoconazole 250 g/l (Score 250 SC 0.05%), copper hydroxide with 50% metallic Cu
(Champ 77 WG 0.25%), metiram 80% (Polygram DF 0.2%), dimethomorph 9% + mancozeb 60% (Acrobat MZ 69 WG
0.2%), mefenoxam 4% + macozeb 64% (Ridomil Gold MZ 68 WG 0.25%), mancozeb 80% (Dithane M 45 WP 0.2%).
Foliar fertilizers have also been used Crop Max 0.3%, Agroleaf Power Total (20.20.20) 0.5% and Agroleaf Power HK
(15.10.31) 0.5%.
Key words: pathogens, Pseudomonas syringae pv. tomato, Xanthomonas campestris pv. Vesicatoria.
INTRODUCTION influence on some seed indicators (Cristea et

al., 2013).
From the vegetable species, tomatoes A particular aspect of the attack of Alternaria
(Lycopersicon esculentum Mill.) represent the spp. is the pathogenic-enzymatic interrelation
largest area of culture. between plant species (Cozea et al., 2011).
Thus, in 2014, tomatoes occupied an area of 5 Pathogens responsible for the occurrence of
million hectares worldwide, with an average tomatoes diseases and their description are
production of 33,988 t/ha (FAO, 2014). presented by Docea et al. (2012) and
Alternaria spp. colonizes different plant species Gheorghies et al. (2001).
with unfavorable effects on production, both For tomato attack (Phytophthora infestans) in
quantitatively and qualitatively (Cristea, 2005). field conditions, prognosis and warning
An important incidence of Alternaria spp. was measures are recommended (Gheorghies et al.,
also reported on the seeds of certain species of 2001). Pathogens attacking tomato crops cause
crop plants (Cristea et al., 2008; Cristea considerable economic damage, which can be
(Manole) et al., 2015; Dudoiu et al., 2016; direct (quantitative reduction of the harvest and
Gruia et al., 2016; Manole (Cristea) et al., damage to its quality) or indirect (social or
2015; Mardare et al., 2014; Pana et al., 2014). economic effects - import from other countries;
Research on the influence of abiotic factors on Severin et al., 2001).
the biological parameters of fungi belonging to In tomato field crops, the following pathogens
the genus Alternaria spp. (Mardare et al., 2015; are frequently attacked: Xanthomonas cam-
Radu et al., 2011), was also carried out, the pestris pv. vesicatoria (staining the leaves and
distribution of these fungi, depending on the blistering the fruits), Pseudomonas syringae
level of the attack (Berca et al., 2015) and their pv. tomato (pustular fruit stain), Alternaria
499
porri f. sp. solani (brown spotting of the leaves crop failure if adequate control measures are
or alternarioza), Fulvia fulva (brown hair not taken (Costache et al., 2007).
staining) and Phytophthora infestans (hand) For the control of these pathogens, products
(Mandru et al., 2017). with different active substances are frequently
Alternaria porri f. sp. solani can cause signify- used: copper hydroxide 50% (Copper Max 50
cant economic damage, the main symptoms WP 0.25%), chlorothalonil 500 g/l (Bravo 500
being collar rot in the basal part of the SC 0.2%), azoxystrobin 200g/l-difenoconazole
seedlings, leaf and stem stains and rotting fruit 125g/l (Ortiva Top 0.1%), iprovalicarb 8.4% +
(Walker, 1952). Cu of oxychloride 40% (Melody Compact 49
The reported production losses can reach 79% WG 0.2%), difenoconazole 250 g/l (Score 250
and have been reported in Canada, India, USA, SC 0.05%), copper hydroxide with 50%
Nigeria (Basu., 1974; Datar et al., 1981; Sherf metallic Cu (Champ 77 WG 0.25%), metiram
et al., 1986; Gwary et al., 1998). 80% (Polygram DF 0.2%), dimethomorph 9%
"Collar" rot occurs at a frequency of 20-40% in + mancozeb 60% (Acrobat MZ 69 WG 0.2%),
seedlings after planting in the field (Sherf et al., mefenoxam 4% + mancozeb 64% (Ridomil
1986). Gold MZ 68 WG 0.25%), mancozeb 80%
Fulvia fulva only attacks the plant foliage, but (Dithane M 45 WP 0.2%).
in favorable conditions it may cause premature The research undertaken at RDIVFG Vidra, in
defoliation (Babadoost, 2011). 2017, aimed to establish treatments for the
Attack of bacteria Xanthomonas campestris pv. simultaneous control of pathogens present in
vesicatoria and Pseudomonas syringae pv. tomato field crops.
tomato is favored by high atmospheric humi-
dity, and is manifested on leaves and fruits, on MATERIALS AND METHODS
petiole, on stems, and can also cause plant
defoliation and fruit degradation (Dafna Tamir- During 2017, at the RDIFG Vidra, it was
Ariel et al., 2007). organized a bifactorial experience, placed on
The most dangerous attack is caused by the subdivision parcels method, with 12
Phytophthora infesters which, under favorable variants and 4 repetitions, using Pontica 102
conditions (moderate temperatures, maximum tomato variety.
atmospheric humidity, the presence of drops of Treatment variants including foliar fertilizers
water on the foliage and fruits) may lead to are presented in Table 1.
Table 1. Experimental variants

V Foliar Phytosanitary June July August
fertilizers tratments Treatments 1, 2 Treatments 3, 4 Treatments 5, 6
1. Copper Max 50 WP 0.25% 3. Ortiva Top 0.1% 5. Ortiva Top 0.1%
1. B1 2. Bravo 500 SC 0.2% 4. Melody Compact 49 WG 0.2% + 6. Melody Compact 49 WG 0.2%
Score 250 SC 0.05%
1. Champ 77 WG 0.25% 3. Ortiva Top 0.1% 5. Ortiva Top 0.1%
2. A1 B2 2. Polyram DF 0.2% 4. Acrobat MZ 69 WG 0.2%+ Score 6. Acrobat MZ 69 WG 0.2%
Crop Max 0.3% 250 SC 0.05%
3. B3 2. Dithane M 45 WP 0.2% 4. Ridomil Gold MZ 68 WG 0.25%+ 6. Ridomil Gold MZ 68 WG 0.25%
Score 250 SC 0.05%
4. B4 Ut. Ut. Ut.
5. B1 2. Bravo 500 SC 0.2% 4. Melody Compact 49 WG 0.2% + 6. Melody Compact 49 WG 0.2%
A2 Score 250 SC 0.05%
Agroleaf Power
Total (20.20.20)
6. B2 2. Polyram DF 0.2% 4. Acrobat MZ 69 WG 0.2%+ Score 6. Acrobat MZ 69 WG 0.2%
0.5% +
250 SC 0.05%
Agroleaf Power
HK (15.10.31)
0.5%
Score 250 SC 0.05%
8. B4 Ut. Ut. Ut.
1.Copper Max 50 WP 0.25% 3. Ortiva Top 0.1% 5. Ortiva Top 0.1%
9. B1 2. Bravo 500 SC 0.2%) 4. Melody Compact 49 WG 0.2% + 6. Melody Compact 49 WG 0.2%
Score 250 SC 0.05%
A3
10. B2 2. Polyram DF 0.2% 4. Acrobat MZ 69 WG 0.2%+ Score 6. Acrobat MZ 69 WG 0.2%
(Untreated
250 SC 0.05%
control )
Score 250 SC 0.05%
12. B4 Ut. Ut. Ut.
500
To reduce the losses caused by the attack of The earliest attack of Pseudomonas syringae
pathogens, the following fungicides– pv. tomato (29.05.) was followed by
bactericides alone or in combination: copper Xanthomonas campestris pv. vesicatoria
hydroxide 50% (Copper Max 50 WP 0.25%), (8.06.), Alternaria porri f. sp. solani (13.06.),
chlorothalonil 500 g/l (Bravo 500 SC 0.2%), Fulvia fulva (16.06.), and Phytophthora
azoxystrobin 200 g/l + difenoconazole 125 g/l infestans (20.06.).
(Ortiva Top 0.1%), iprovalicarb 8.4 % + Cu of The rise and evolution of the attack was
oxychloride 40 % (Melody Compact 49 WG favored by rainfall in May (71.0 mm), June
0.2%), difenoconazole 250 g/l (Score 250 SC (43.5 mm) and July (99.0 mm) and maximum
0.05%), copper hydroxide with 50% metallic atmospheric humidity of over 70%, so at the
Cu (Champ 77 WG 0.25%), metiram 80% end the third decade of August recorded values
(Polyram DF 0.2%), dimethomorph 9% + between 8.4% (Fulvia fulva) and 13.7%
mancozeb 60% (Acrobat MZ 69 WG 0.2%), (Phytophthora infestans - Table 2).
mefenoxam 4% + mancozeb 64% (Ridomil Mandru et al. (2017) also carried out research
Gold MZ 68 WG 0.25%), mancozeb 80% on tomato culture, which identified the
(Dithane M 45 WP 0.2%). following pathogens on the foliage:
These have been established according to the Pseudomonas syringae pv. tomato, Alternaria
sequence of pathogens, and the interval bet- porri f. sp. solani, Fulvia fulva and
ween them in correlation with climatic factors. Phytophthora infestans.
There were applied 6 foliar treatments at Symptoms produced by the pathogens on the
intervals of 8-17 days. tomato plant foliage are shown in Figures 1, 2,
Foliar fertilizers were used: Crop Max 0.3% (4 3, 4 and 5.
foliar treatments at 10 days intervals), Agroleaf
Power Total (20.20.20) 0.5% (3 foliar
treatments at 10 days intervals) and Agroleaf
Power HK (15.10.31) 0.5% (one treatment after
Agroleaf Power Total).
Dynamic observations have been made on the
occurrence and evolution of pathogen attack
(frequency and severity of the attack) in
relation to climatic factors.
The best treatment variants have been
established according to the average efficacy
and the obtained production.
Qualitative determinations for total dry
substance, soluble dry matter, carbohydrate Figure 1. Attack by Pseudomonas syringae pv. tomato
content and vitamin C in tomato fruits from on the foliage
variants A1B4 (fertilized with Crop Max foliar
0.3%), A2B4 (fertilized with Agroleaf Power
Foliar Total (20.20.20) 0.5% + Agroleaf Power
HK (15.10.31) 0.5% and A3B4 (untreated
control).
During 2017, in the Vidra area, the fall -

summer tomato crops, the Pontica variety 102,
the following pathogen attack could be seen:
Pseudomonas syringae pv. tomato,
Xanthomonas campestris pv. vesicatoria,
Alternaria porri f. sp. solani, Fulvia fulva and Figure 2. Attack by Xanthomonas campestris pv.
Phytophthora infestans. vesicatoria on the foliage
501
Figure 3. Attack by Alternaria porri f. sp.solani Figure 4. Attack by Fulvia fulva
on the foliage on the foliage
a) on foliage b) on leaves
Figure 5. Attack by Phytophthora infestans
Table 2. Influence of climatic factors on the occurrence and evolution of pathogen attack
to the tomato field crop (Vidra, 2017)
The degree of attack/month/decade
Pathogenic agents Date of
May June July August
and climatic factors the attack
I II III I II III I II III I II III
Pseudomonas syringae pv. tomato 29.05 0 0 0.2 0.5 3.3 5.2 5.8 6.4 7.9 8.5 10.1 12.2
Xanthomonas campestris pv. 8.06 0 0 0 0.3 2.7 4.6 5.2 5.8 6.7 7.0 7.9 9.5
vesicatoria
Alternaria porri .f. sp. solani 13.06 0 0 0 0 0.7 4.9 5.5 7.3 8.1 10.0 11.3 12.7
Fulvia fulva 16.06 0 0 0 0 0.2 2.8 3.5 4.8 6.0 6.8 7.7 8.4
Phytophthora infestans 20.06 0 0 0 0 0.7 5.8 7.5 8.5 9.6 10.3 12.2 13.7
Temperature minimum (0C) - 10.9 10.5 12.4 15.0 14.0 16.3 15.9 15.9 17.5 21.1 18.3 15.1
Temperature average (0C) - 14.7 15.3 16.5 20.1 19.8 23.8 22.0 22.0 24.0 30.8 25.6 21.3
Temperature maximum (0C) - 19.4 20.9 21.4 27.0 26.0 31.9 28.4 28.9 31.0 36.6 31.8 28.7
Minimum relative humidity (%) - 60.2 53.7 57.0 47.5 43.8 36.4 46.0 37.8 35.7 26.9 27.4 30.2
Average relative humidity (%) - 70,3 63.1 66.9 59.5 56.8 56.4 57.9 47.6 46.8 36.7 38.7 41.7
Maximum relative humidity (%) - 85.2 76.3 76.6 77.8 77.4 82.5 74.7 63.6 63.6 52.4 57.0 56.4
Precipitation (mm) - 43.6 19.9 7.5 20.0 22.5 1.0 84.0 8.5 6.5 0 0 45.0
All variants of experiments have shown good Phytophthora infestans (Table 3). Among them
results in controlling the pathogens were A3B1 (E = 88.3%), A3B3 (E = 86.3%)
Pseudomonas syringae pv. tomato, followed by A1B1 (E = 87.4%), A1B3 (E =
Xanthomonas campestris pv. vesicatoria, 85.3%), A2B1 (E = 86.3%) and A2B3 (E =
Alternaria porri f. sp. solani, Fulvia fulva and 84.9%).
502
Table 3. Influence of phytosanitary treatments and foliar fertilizers on the attack of pathogens
on field tomato culture (Vidra, 2017)
The degree of attack on the foil (%)
Pseudomonas Xanthomonas Alternaria porri f. sp. Fulvia fulva Phytophthora Total Effectiveness
V
syringae pv. campestris pv. solani infestans (%)
tomato vesicatoria
1. 1.3 1.2 1.1 1.8 1.7 7.1 87.4
2. 1.7 1.5 1.3 1.9 2.2 8.6 84.8
3. 1.5 1.4 1.4 1.5 2.5 8.3 85.3
4. 12.2 9.5 12.7 8.4 13.7 56.5 -
5. 1.5 1.4 1.4 2.0 2.4 8.7 86.3
6. 1.9 1.7 1.5 2.1 2.9 10.1 84.1
7. 1.8 1.6 1.7 1.7 2.8 9.6 84.9
8. 13.7 10.7 14.1 9.9 15.2 63.6 -
9. 1.0 0.9 0.8 1.5 1.9 6.1 88.3
10. 1.5 1.3 1.1 1.7 2.4 8.0 84.6
11. 1.2 1.1 1.2 1.3 2.3 7.1 86.3(2)
12. 11.6 8.7 11.5 8.0 12.2 52.0 -
Good results were observed at variant B1 Research on the control of pathogens

variants with 6.13 kg/m2 (134.1%) and B3 with Alternaria porri f. sp. solani, Botrytis cinerea,
6.04 kg/m2 (132.2%, Table 4) compared with Fulvia fulva and Phytophthora infestans in
B4 (the control untreated) at which production tomato crops were also carried out by Costache
was 4.57 kg/m2. Good results were also et al. (2017), which established the efficacy and
obtained in variant B2 with 5.96 kg/m2. influence on the production of combinations of
Regarding the differences in production fungicides in simultaneous control thereof.
obtained in addition to the untreated control
variant, these are very significant in all three
Table 4. Influence of phytosanitary treatments on production (Vidra, 2017)

Factor Production
B kg/m2 (%) The difference from untreated varian Signification
B1 6.13 134.1 +1.56 ***
B2 5.96 130.4 +1.39 ***
B3 6.04 132.2 +1.47 ***
B4 4.57 100.0 - -
DL 5%=0.021; DL 1%=0.029; DL 0.1%=0.039
The analysis of the data presented in Table 5 significantly influence the quantity of the
shows that treatments with foliar fertilizers (A1 obtained production, the values being very
fertilized with Crop Max 0.3%, A2 fertilized close: at A1 5.70 kg/m2 was obtained, at A2
with Agroleaf Power Total (20.20.20) 0.5% and 5.67 k/m2 and at A3 (Mt) 5.65 kg/m2.
Agroleaf Power HK (15.10.31) 0.5% did not
Table 5. Influence of foliar fertilizer treatments on production (Vidra, 2017)
Factor A Production
kg/m2 (%) The difference from untreated varian Signification
A1 5.70 100.88 + 0.05 -
A2 5.67 100.35 + 0.02 -
A3 (Ut.) 5.65 100.00 - -
DL 5%=0.070; DL 1%=0.100; DL 0.1%=0.172
Analyzing the data presented in Table 6, it was Among the parameters analyzed for
found that in all cases, in the variants treated (1, determining the influence of foliar treatments
2, 3, 5, 6, 7, 9, 10, 11), the differences in on Crop Max 0.3%, Agroleaf Power Total
production obtained in addition by the (20.20.20) 0.5% and Agroleaf Power HK
untreated witness (4, 8, 12) are very significant. (15.10.31) 0.5%, it was found that compared to
the untreated control without foliar treatments,
503
fruit content in total dry substance (TDS), Thus, the total dry fruit content of the fruit was
soluble dry matter (SDM), carbohydrate higher by 13.4-15.1%, the dry substance
content and vitamin C content were clearly content soluble by 33.3-44.4%, the
influenced (Table 7). carbohydrate content by 37.8-44.4%, and the
vitamin C content by 8.3-29.2%.
Table 6. Influence of phytosanitary treatments and foliar fertilizers on production (Vidra, 2017)
Production
Foliar Phytosanitary
Variants The difference from
fertilizers tratments kg/m2 % Signification
untreated variant
1. A1 B1 6.15 134.6 +1.58 ***
2. B2 6.02 131.7 +1.45 ***
3. B3 6.07 132.8 +1.50 ***
4. B4 (Ut.) 4.57 100.0 - -
5. A2 B1 6.12 132.5 +1.50 ***
6. B2 5.90 127.7 +1.28 ***
7. B3 6.05 130.9 +1.43 ***
8. B4 (Ut.) 4.62 100.0 - -
9. A3 (Ut.) B1 6.13 135.6 +1.61 ***
10. B2 5.96 131.9 +1.44 ***
11. B3 6.01 132.9 +1.49 ***
12. B4 (Ut.) 4.52 100.0 - -
DL 5%=0.065; DL 1%=0.087; DL 0,1%=0.110
Table 7. Influence of foliar fertilizers on the quality of tomato fruits

Vitamin
Acidity
C
Variation of (g citric
V. Water (%) TDS (%) SDM (%) (%) Glucids (%) (mg/ (%)
fertilization acid at
100g
100 g s.p).
s.p)
1. Crop Max 0.3% 93.56 99.2 6.44 113.4 3.9 144.5 0.77 100.0 3.25 144.4 27.28 129.2
Agroleaf Power
Total (20.20.20)
2. 0.5% + Agroleaf 93.46 99.1 6.54 115.1 3.6 133.3 0.70 90.9 3.10 137.8 22.88 108.3
Power HK
(15.10.31) 0.5%
3. Untreated control 94.32 100.0 5.68 100.0 2.7 100.0 0.77 100.0 2.25 100.0 21.12 100.0
CONCLUSIONS Gold MZ 68 WG 0.25%) with production of

6.04 kg/m2 (132.2%; E medium = 85.5%).
In the field tomato crops, the pathogens Treatments with foliar fertilizers Crop Max
Pseudomonas syringae pv. tomato, 0.3%, Agroleaf Power Total (20.20.20) 0.5%
Xanthomonas campestris pv. vesication,, and Agroleaf Power HK (15.10.31) 0.5% did
Alternaria porri f. sp. solani, Fulvia fulva and not significantly influence production in terms
Phytophthora infestans diminish production in of quantity but only qualitatively: the total dry
terms of quantity and qualitatively impair it. substance (TSS) increase by 13.4-15.1%, the
Among the variants of experienced treatments dry substance soluble (DSS) by 33.3-44.4%,
were B1 (T1: Copper Max 50 WP 0.25%; T2: the carbohydrate content by 37.8-44.4% and
Bravo 500 SC 0.2%; T3: Ortiva Top 0.1%; T4: the vitamin C content with 8.3-29.2%.
Melody Compact 49 WG 0.2% + Score 250 SC
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506
EXOGENOUS CYTOKININ APPLICATION INCREASED

THE CAPSAICIN AND ASCORBIC ACID CONTENT IN PEPPER FRUIT
Mostafakamal SHAMS1, Ertan YILDIRIM1, Melek EKINCI1, Guleray AGAR2,

Metin TURAN3, Raziye KUL1
1
Department of Horticulture, Faculty of Agriculture, Atatürk University, Erzurum, Turkey
2
Department of Biology, Faculty of Science, Atatürk University, Erzurum, Turkey
3
Department of Genetic and Bioengineering, Yeditepe University, Istanbul, Turkey
Abstract
Chili peppers synthesize capsaicin and accumulate it in the fruits. Many biochemical studies have been done in the field
of capsaicin biosynthesis in chili peppers. The levels of capsaicin vary among chili pepper cultivars, and it is also
affected by environmental conditions. The present study assayed the effect of cytokinin on the capsaicin and ascorbic
acid content of Capsicum annum var ʻIlicaʼ (pungent pepper) in the greenhouse. Cytokinin levels include 0, 50 and 100
µM of Benzyl amino purine and the treatments were applied at every 10 days until the harvest beginning 5 days after
planting. The results of this study monitored that cytokinin (benzyl amino purine) treatment increased the capsaicin and
ascorbic acid content in pepper fruits as compared to the control but decreased the fruit number per plant.
Key words: ascorbic acid, capsaicin, fruit, pungent pepper.
INTRODUCTION application of nitrate can increase the capsaicin

Hot pepper burning sensation induced by the biosynthesis, but potassium application had not
presence of acid amides, collectively known as a significant effect on it. Ascorbic acid is very
capsaicinoids, that are formed from abundant in pepper fruits and is the main
phenylalanine and valine or leucine. Capsaicin component of vitamin C. Its content is highly
and dihydro indicate more than 90% of the total varying among cultivar and ripening stage (Bae
capsaicinoid content in most pungent peppers et al., 2014).
(Ben-Chaim et al., 2006). Capsaicinoid Cytokinin is a hormone of plants that adjusts
biosynthesis is restricted to the genus Capsicum growth and developmental cycles. It has an
(Stewart et al., 2007). Capsaicin content raises important role on the chlorophyll status in
slowly during fruit development reaching peak plants (Lim et al., 2007). Exogenous treatment
levels at 40 to 50 days after planting of N6-benzyl amino purine (BAP) on the
(Contreras-Padilla and Yahia, 1998), after eggplant increased its tolerance under salt
which it gravitates to deteriorate into secondary stress by decreasing the O 2- and
compounds due to peroxidase action (Bernal malondialdehyde production rate (Wu et al.,
and Barceló, 1996). For increasing or 2014). Application of BAP in the in vitro
improving pungent compound production, a culture increased the comptothecin content in
research has revealed that hydric stress shoots and roots of Ophiorrhiza rugosa var.
ameliorates capsaicinoid levels, because water decumbens (Vineesh et al., 2007). It was
deficit affects the phenylpropanoid pathway reported that the application of BAP increased
(Estrada et al., 1999). the shikonin formation in Onosma paniculatum
Hydric stress can raise capsaicin levels by cultured cell (Ding et al., 2004). So BAP has an
increasing activity of the enzymes exclusive effect on the plant growth and
phenylalanine ammonia-lyase (PAL), cinnamic secondary metabolites production in the plants.
acid-4-hydroxylase (C4H), and capsaicinoid The results of the literature review suggest that
synthetases (CS), all engaged in capsaicin abiotic and biotic parameters could affect the
biosynthesis (Sung et al., 2005). secondary metabolites production in plants. Up
MonforteǦGonzález et al. (2010) found that to now a research has not been done on the
507
effect of cytokinin on the capsaicin production, Ascorbic acid analysis
so in this study, we investigated the effect of Five grams of well-homogenised sample were
BAP on the capsaicin and ascorbic acid content disrupted in a crucible mortar with quartz sand.
in pepper. To the macerate 50 mL of meta phosphoric acid
(analytical grade) was gradually added and the
MATERIALS AND METHODS mixture was then transferred to a 100 mL
The experiment was performed in the Ataturk Erlenmeyer flask, closed with stopper and then
University under controlled greenhouse filtered. The filtrate was purified in addition by
conditions. Seeds of Capsicum annum var. Ilica passing through a 0.45 mm PTFE syringe filter
(pungent pepper) were germinated on the 86- before injection on HPLC column.
celled styrofoam trays filled with peat, then the The analytical determination of ascorbic acid
homogenous and healthy seedlings were was performed on C18 Nautilus, 100-5, 150 ×
transplanted to the pots in size 30 x 20 x 20 cm 4.6 mm column with gradient elution of 0.01 M
after thirty days. The pots were filled with a KH2PO4 (A) and acetonitrile (B). The gradient
ratio of 2: 1: 1 (v: v: v) of soil: sand: manure elution started with 1% B in A and changed to
having around 1.30 g cm3 bulk density. 30% B in A in 15 min.; then, it turned to 1% A
The greenhouse mean temperature and relative in B in 5 min. The flow rate was 0.7 mL/min.
humidity were 24°C and 74%, respectively The highest absorption maximum of ascorbic
during the experiment. The cytokinin treatment acid under these conditions was detected at
included 0, 50 and 100 µM of N6-benzyl amino 265 nm. For quantitative determination of
purine (BAP). The BAP solutions were ascorbic acid, standard materials (Sigma-
prepared with distilled water containing 0.02% Aldrich, Budapest, Hungary) were used.
Tween 20 as a surfactant, and the control Stock solutions and then working solutions
(without BAP) treatment just was contained were prepared for each compound to make the
distilled water and surfactant. Five days after calibration between concentration and peak
seedling transplanting, the BAP solutions were area (Nagy et al., 2015).
sprayed to leaves in the afternoon and the foliar Data analysis
spraying continued every 10 days until the A completely randomized experimental design
harvest. Half-strength Hoagland nutrient solu- was used in this study. Each treatment had 4
tion was used with irrigation (Rubio et al., replications with 5 plants for each replication.
2011). Statistical analysis was carried out by SPSS
Marketable fruits were selectively harvested version19 at P≤ 0.001, the mean separation was
weekly from 55 days after transplanting until done following Duncan’s Multiple Range Test.
the end of the experiment. In addition, at the
end of the experiment, one representative RESULTS AND DISCUSSIONS
marketable pepper fruit (18 fruits) was selected
from different plants and used for determining As shown in the Figure 1, BAP treatments
chemical analysis. Yield was determined by significantly increased the capsaicin content in
counting and weighing all fruits on each plant. fruits of pepper. Treatment of BAP (100 µM)
Marketable fruit yield was determined had the highest effect on the capsaicin content,
according to the color, the health state, the as compared to the untreated ones. Exogenous
shape, and the weight. BAP treatment had a significant effect on the
Capsaicin analysis ascorbic acid content in the pepper fruits, by
The samples were separated in a SHIM-PACK increasing the BAP concentration its content
VP-ODS column (150 mm × 4.6 mm; enhanced. Higher concentration of BAP (100
Shimazu). For capsaicin, the eluent was a µM) increased the ascorbic acid content 22% in
mixture of acetonitrile/1% acetic acid (2:1 v/v), comparison to the untreated one (Figure 2). The
and the flow rate was 1 ml/min. All eluates cytokinin (BAP) application had not a positive
were monitored at 280 nm using a UV detector. effect on the fruit number in pepper plant. The
External standards were prepared by dissolving BAP treatment (100 µM) decreased the fruit
commercial capsaicin (Sigma-Aldrich) in number but its lower concentration (50 µM)
methanol and acetonitrile (Ogawa et al., 2015).
508
had not a significant effect as compared to the
control plants (Figure 3).
Previous studies have shown that capsaicin 100 A
biosynthesis exclusively is occurred in the fruit 90
80
(Stewart et al., 2005; Stewart et al., 2007). 70 B
Fruit number plant-1

Cytokinin has an important role in the 60 B
adjustment of different biological processes, 50
involving growth and development, 40
furthermore, acclimation/adaptation to 30
environmental conditions in plants. Li et al. 20
10
(2016) reported that cytokinin application 0
increased secondary metabolite production in 0 50 100
Morinda citrifolia. BAP Levels (µM)
Figure 3. Effect of N6-benzyl amino purine (BAP) levels

on number of fruit in pepper. Data followed by a
Capsaicin content (mg.kg -1)
35 different letter were significantly different (P≤ 0.001)

A
30 according to the Duncan Multiple Range Test
B
25 C
Similarly, Govindaraju and Indra Arulselvi
20 (2016) reported that BAP treatment enhanced
15 the PAL and subsequently secondary
10 metabolites production on in vitro media in
5 Coleus aromaticus, without changes in plant
0 genetic. In this study, exogenously BAP
0 50 100 application increased the capsaicin content in
BAP Levels (µM) fruits. It could be due to role of cytokinin on
growth and development including meristem
Figure 1. Effect of N6-benzyl amino purine (BAP) levels maintenance, vascular development, modu-
on capsaicin content in fruits of pepper. Data followed lation of sink-source relationships and nutrient
by a different letter were significantly different (P≤ acquisition in roots.
0.001) according to the Duncan Multiple Range Test BAP also play an important role in gene
expression in capsaicin production. Moreover,
Govindaraju and Indra Arulselvi (2016)
Ascorbic acid content (ߤg mL-1)
70 A reported that BAP application on in vitro

60
propagation of Coleus aromaticus increased
B PAL gene expression, PAL directs
50 C
phenylalanine (Phe) to secondary metabolism,
40 deamination this amino acid to generate
30 cinnamic acid, that could be converted to
20 various phenolic compounds, involving
10
capsaicin (Castro-Concha et al., 2016).
The ascorbic acid content with a gradual
0
increase in fruit ripening was detected in hot
0 50 100 pepper (Iqbal et al., 2013). Navarro et al.
BAP Levels (µM)
(2006) revealed that salinity decreased ascorbic
acid content in pepper and methyl jasmonate
Figure 2. Effect of N6-benzyl amino purine (BAP) levels application increased antioxidant capacity of
on ascorbic acid content in fruits of pepper. Data strawberry fruits (Ayala-Zavala et al., 2005). In
followed by a different letter were significantly different
(P≤ 0.001) according to the Duncan Multiple Range Test this study, BAP treatment increased ascorbic
acid in Ilica pepper. It means that cytokinins
509
have a role in ascorbic acid biosynthesis in pepper. Journal of Agricultural and Food Chemistry,
pepper plants. 44 (10): 3085-3089.
Contreras-Padilla M., Yahia E.M., 1998. Changes in
The previous study have been shown that Capsaicinoids during Development, Maturation, and
cytokinin application had not a significant Senescence of Chilli Peppers and Relation with
effect on the yield and dry matter accumulation Peroxidase Activity. Journal of Agricultural and Food
in the ryegrass (Lolium perenne L.) and white Chemistry, 46: 2075−2079.
clover (Trifolium repens L.) (Ghani et al., Castro-Concha L., Baas-Espinola F.M., Ancona-
Escalante W.R., Vázquez-Flota F.A., Miranda-
2014). However, we found that BAP had a HamEmail M.L., 2016. Phenylalanine biosynthesis
significant effect on the fruit number. In this and its relationship to accumulation of capsaicinoids
regard, the cytokinin higher levels (50 and 100 during Capsicum chinense fruit development.
µM of BAP) decreased the fruit number of Biologia Plantarum, 60 (3): 579-584. DOI:
pepper. The decline of fruit/plant could be due https://doi.org/10.1007/s10535-016-0608-4.
Ding J., Shi S., Jiang B.H., Yang Y.H., Huang J., Shen
to its role in regeneration of shoot which it H.G., Xia K., Zhang J., Jiang X., 2004. Effects of
leads to the shadowing of plants on each other methyl jasmonate with indole-3-acetic acid and 6-
and it ultimately causes to decrease in the enzylaminopurine on the secondary metabolism of
plants yield. cultured Onosma paniculatum cells. In Vitro Cellular
& Developmental Biology - Plant, 40 (6): 581-585.
DOI: https://doi.org/10.1079/IVP2004578.
CONCLUSIONS Estrada B., Pomar F., Dı́az J., Merino F., Bernal M.A.,
1999. Pungency level in fruits of the Padrón pepper
The literature review shows that capsaicin is
with different water supply. Scientia Horticulturae, 81
responsible for pungent sensation in pepper (4): 385-396.
cultivators and the fruit placenta is a main place Ghani A., Ledgard S., Wyati J., Catto. W., 2014.
for capsaicin production. BAP treatment had a Agronomic assessment of gibberellic acid and
significantly effect on capsaicin (CAP) cytokinin plant growth regulators with nitrogen
fertiliser application for increasing dry matter
production and increased CAP and ascorbic
production and reducing the environmental footprint.
acid content in pepper. Proceedings of the New Zeeland Grassland
Association. 76: 177-182.
Govindaraju S., Arulselvi P.I., 2016. Effect of cytokinin
ACKNOWLEDGMENTS combined elicitors (l-phenylalanine, salicylic acid and
chitosan) on in vitro propagation, secondary
The authors acknowledge Ataturk University metabolites and molecular characterization of
for generous financial support. medicinal herb - Coleus aromaticus Benth (L). J
Saudi Soc Agric Sci., DOI
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MonforteǦGonzález, M., Guzmán-Antonio A., Uuh-Chim
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511
512
ESSENTIAL OILS AND HOMEMADE FUNGICIDES AGAINST FUSARIUM

OXYSPORUM F. SP. LYCOPERSICI TOMATO PATHOGEN
Andreea-Alexandra TIMOFTE, Andreea-Mihaela FLOREA, Florin Daniel LIPȘA,

Eugen ULEA
„Ion Ionescu de la Brad” University of Agricultural Sciences and Veterinary Medicine of Iasi,
3 Mihail Sadoveanu Alley, 700490, Romania
Abstract
The destructive disease of tomato worldwide, fusarium wilt, is caused by Fusarium oxysporum f. sp. lycopersici (Sacc)
W. C. Synder & H. N. Hans., a vascular wilt pathogen. The fungus affects greenhouse and field grown tomatoes in
warm vegetable production areas. Yellowed leaves and wilted plants with low or absent crop yield are the main
symptoms of the disease. In this paper we have analyzed possible substitutes to synthetic fungicides in controlling the
pathogen. Since the potential of essential oils as antimicrobial agents is well established and farmers around the world
already use traditional recipes, we have decided to test a fair amount of essential oils and four homemade fungicides
against the growth of Fusarium oxysporum f. sp. lycopersici mycelium. For the experiment, 1 ml of essential oil serial
dilutions of 0.1%, 1% and 10% were used along with the most used worldwide homemade fungicides. Fungal growth
measurements were taken every 24h for 13 days, until no fungal growth was registered. Results show that 0.1%
essential oils concentration had no inhibitory effect, while at 1% oil concentration only oregano essential oil was 61%
effective. At 10% cinnamon, cloves, thyme, oregano and lemon essential oils showed cidal effects.
Key words: essential oils, fusarium wilt, in vitro, tomato disease.
INTRODUCTION production areas. Yellowed leaves and wilted

plants with low or absent crop yield are the
The unifying principle behind most supporters main symptoms of the disease (Sudhamoy et
of organic farming is the belief that soil health al., 2009). The disease may lead to a 30 to 40%
is extremely important for life and is the only yield loss and it can even go up to 80% under
sustainable way of cultivating land and creating favorable climate conditions (Kirankumar et
a safe future for humanity. al., 2008).
The use of biopesticides has been possible The pathogen enters the root and further into
since the 1960s, but has only increased recently the vascular tissue. The xylem vessels are
due to society's awareness of the issues of colonized by mycelium and conidia. As a
intensive use of chemicals such as human result, the vessels are clogged and plants suffer
health, soil pollution and water resources, such of severe water stress, leading to the
as and the resistance acquired by pests and characteristic wilt symptoms (Beckman, 1987).
pathogens. Biocontrol products are used in both There are three physiological pathogen races
conventional and organic farming. (1, 2 and 3) known to affect tomato cultivars
Tomatoes (Lycopersicon esculentum Mill.) are (Kawabe et al., 2005).
highly susceptible to pathogens, such as fungi,
viruses, bacteria and nematodes, which cause MATERIALS AND METHODS
severe yield losses (Barone and Frusciante,
2007). The antifungal activity of 22 commercially
The destructive disease of tomato worldwide, purchased essential oils, i.e. anise (Pimpinella
fusarium wilt is caused by Fusarium anisum L.), basil (Ocimum basilicum L.),
oxysporum f. sp. lycopersici (Sacc) W. C. Indian frankincense (Boswellia serrata T.),
Synder & H. N. Hans., a vascular wilt cinnamon (Cinnamomum aromaticum L.),
pathogen. The fungus affects greenhouse and camphor tree (Cinnamomum camphora L.),
field grown tomatoes in warm vegetable lemongrass (Cymbopogon winterianus L.),
513
cloves (Syzygium aromaticum L.), coriander Fungal growth measurements were taken every
(Coriandrum sativum L.), May Chang (Litsea 24 h for 13 days, until no fungal growth was
cubeba), fennel (Foeniculum vulgare M.), oil registered. One Way Within Subjects ANOVA
grass (Cymbopogon citratus DC.), lavender was used to determine the effect of treatments
(Lavandula angustifolia Mill.), tea tree on each concentration on growth
(Melaleuca viridiflora Sol.), orange (Citrus x measurements. Statistical analysis was
sinensis L.), palmarosa (Cymbopogon martinii performed with the IBM SPSS Amos v20.
Roxb.), turmeric (Curcuma longa), rosemary
(Rosmarinus officinalis L.), clary sage (Salvia RESULTS AND DISCUSSIONS
sclarea L.), spearmint (Mentha spicata L.),
thyme (Thymus vulgaris L.), oregano The maximum growth of F. oxysporum f. sp.
(Origanum vulgare L.) and lemon (Citrus lycopersici at the end of the 13 days experiment
limon L.), along with homemade fungicides is outlined in Figure 1.
based on baking soda, garlic and hydrogen
perodixe were investigated against F.
oxysporum f. sp. lycopersici.
For the homemade fungicides, we have selected
the most used recipes by international farmers:
mix 5 tablespoons of baking soda with 1
teaspoon of liquid soap in 1 gallon of water
(B1); mix 1 tablespoon of baking soda with 1
teaspoon of castor oil in 1 gallon of water (B2);
chop 100 g of garlic cloves in 1 L of water Us
(Ga); hydrogen peroxide 3% AO (HP).
For the experiment, 1 mL of EO dilution or
homemade fungicide was pipetted in each Petri
dish (85mm), after which 17 mL of potato Figure 1. Effect of EOs and homemade fungicides on
dextrose agar (PDA, Scharlau, Spain) were growth of Fusarium oxysporum f. sp. lycopersici, at
added at temperature of 50ºC, to avoid different concentrations (original)
volatilizing or denaturing the aromatic
compounds in the oils, then the dishes were The oregano (Origanum vulgare) EO is the
stirred for 20s. EO concentrations of 0.1%, 1% only oil that has the most aggressive effect on
and 10% were expressed using Percent the growth of the fungus: both at 1% and 10%
Composition by Mass (%), in which the mass concentration, the mycelium was stopped from
of the solute is divided by the mass of the developing (Figure 2); on the other hand, the
solution (mass of solute plus mass of solvent), 0.1% oil concentration did not prevent growth
then multiplied by 100. Media was allowed to in any way. The main components of the oil are
cool and solidify. After 2 h, a 7 mm mycelial carvacrol, thymol, p-cymene, cis-omen,
plug of F. oxysporum f. sp. lycopersici was caryophyllene and linalool.
centered onto each Petri dish. The F.
oxysporum f. sp. lycopersici cores were taken
from the edge of individual 14 days old
colonies. All Petri dishes were left inside the
laminar-flow hood for 24 h then stored inverted
so that water would not condense on the agar
surface. Dishes were incubated in the dark
inside a 28ºC germination chamber. A total of Figure 2. Cidal effect of oregano EO on growth of
66 dishes were used per replication, with 3 Fusarium oxysporum f. sp. lycopersici, at 1% (a) and
replications. Control plates were included in 10% (b) oil concentrations (original)
each replication: 3 PDA media plates
inoculated with the pathogen, to determine the There are a number of essential oils that did not
viability and growth. allow the fungus any growth at 10% oil
514
concentration, i.e. cinnamon (Cinnamomum aromaticum) and thyme (Thymus vulgaris) EOs at 0.1%,
aromaticum), cloves (Syzygium aromaticum), 1% and 10% concentrations (original)
thyme (Thymus vulgaris) (Figures 3 and 4), and Effects of EOs treatments on growth of F.
lemon (Citratus limon) EOs. oxysporum f. sp. lycopersici are significant,
Wilks’Lambda = 0.71, F (2, 20) = 3.95, p =
0.036 (Table 1).
Table 1. Multivariate Testsa - Fusarium oxysporum f. sp.

lycopersici
Effect Va- F Hypo- Error Sig.
Figure 3. Cidal effect of cinnamon (Cinnamomum
lue thesis df df
aromaticum) (a), cloves (Syzygium aromaticum) (b) and
thyme (Thymus vulgaris) (c) EOs on Fusarium Pillai's Trace .283 3.954b 2.000 20.000 .036
oxysporum f. sp. lycopersici, 10% oil concentration
(original) Factor Wilks' Lambda .717 3.954b 2.000 20.000 .036
1 Hotelling's Trace .395 3.954 b

2.000 20.000 .036
Cinnamon and cloves EOs have eugenol and Roy's Largest Root .395 3.954b 2.000 20.000 .036
eugenol acetate. As distinct elements, a. Design: Intercept
cinnamon oil also contains cinnamic aldehyde Within Subjects Design: factor1
b. Exact statistic
and benzyl benzoate, and cloves oil also has
iso-eugenol and caryophyllene. The α-pinene,
Since the Sig. value for Wilks’ Lambda
camphen, β-pinene, α-terpinene and linalool are
variation is lower than 0.05, there is a
found to be both in thyme and lemon EOs.
significant difference between the effect of
each oil concentration.
The Paired T-Test (Table 2) was further used to
compare the post hoc effect of each oil
concentration on the fungus development.
Figure 4. Average size of Fusarium oxysporum f. sp.

lycopersici mycelium under treatment with cinnamon
(Cinnamomum aromaticum), cloves (Syzygium
Table 2. Paired Samples Test - Fusarium oxysporum f. sp. lycopersici

Paired Differences
Std. 95% Confidence Interval of the Sig.
Std. t df
Mean Error Difference (2-tailed)
Deviation
Mean Lower Upper
concentration=0.1% -
-2.227 13.349 2.846 -8.146 3.691 -.783 21 .443
concentration=1%
concentration=0.1% -
-25.455 41.977 8.950 -44.066 -6.843 -2.844 21 .010
concentration=10%
concentration=1% -
-23.227 38.295 8.164 -40.206 -6.248 -2.845 21 .010
concentration=10%
The T-test indicates there are significant differences between 0.1% - 10%, as well as between 1% -
10% EOs concentrations, since the Sig. value, also known as the p value, is lower than 0.017.
515
All the EOs showed low influences at 0.1% Barone A., Frusciante L., 2007. Marker-assisted
selection: current status and future perspectives in
concentrations, up to 14%. crops, livestock, forestry and fish (eds Guimaraes E.
The oregano (Origanum vulgare) EO treatment et al.) Ch. 9, 151-164 (Agriculture and Consumer
demonstrated a 61% efficacy on the F. oxysporum Protection Dept.).
f. sp. lycopersici mycelium at 1% oil Beckman C.H., 1987. The nature of wilt diseases of
concentration. plants 175-176 (American Phytopathological
Society).
The fungus was influenced by a reduced Hariprasad P., Navya H.M., Chandra Nayaka S.,
number of EOs at 10% concentration, but they Niranjana S.R., 2009. Advantage of PSIRB over
all demonstrated a cidal effect, i.e. cinnamon PSRB and IRB to improve plant health of tomato.
(Cinnamomum aromaticum), cloves (Syzygium Biol. Cont. 50, 307-316.
aromaticum), thyme (Thymus vulgaris), oregano Kawabe M. et al., 2005. Three evolutionary lineages of
(Origanum vulgare) and lemon (Citrus limon). tomato wilt pathogen, Fusarium oxysporum f. sp.
lycopersici, based on sequences of IGS, MAT1, and
Usually, the effects are seen in a time- and
pg1, are each composed of isolates of a single mating
dose-dependent manner; higher concentrations type and a single or closely related vegetative
cause severe effects more rapidly. compatibility group. J. Gen. Plant Pathol. 71, 263-
As the present paper showed, the oil 272.
concentrations necessary to kill F. oxysporum f. Kirankumar R., Jagadeesh K.S., Krishnaraj P.U., Patil
M.S., 2008. Enhanced growth promotion of Tomato
sp. lycopersici are most of the times much
and nutrient uptake by plant growth promoting
higher than those required to inhibit its growth. rhizobacterial isolates in presence of Tobacco Mosaic
Virus pathogen. J. Agric. Sci. 21, 309-311.
Sudhamoy M., Nitupama M., Adinpunya M., 2009.
Salicylic acid induced resistance to Fusarium
oxysporum f. sp. lycopersici in tomato. Plant Physiol.
Biochem. 47, 642-649.
516
POSITIVE ASPECTS OF AN ORNAMENTAL VEGETABLE GARDEN AND

ITS EFFECTS TOWARDS FAMILY AND COMMUNITY SUSTAINABILITY
Florina-Maria GALEA (DELEANU), Neculai MUNTEANU,

Vasile STOLERU, Gabriel Ciprian TELIBAN
3 Mihail Sadoveanu Alley, Iași, Romania
Abstract
Sustainability is based on three important pylons, the environment, the social component and the economic component.
Design based on landscape principles and rules and by applying the correct crop technology, the vegetable garden
presents many functions, which increase its impact on the community and on the environment. To reach the aim and
objectives of this research based on the obtained results during 2015-2017 regarding crop productivity, plant
development, economic aspects and ornamental criteria a survey regarding sustainability of the vegetable garden was
done. From an economic point of view, the studied garden has a positive impact on the family budget. The garden also
has a big impact on the owners bringing benefits to their health and also on the education of the young generation
regarding horticultural practices. The results of the study show that the ornamental vegetable garden, design in an
intercropping system has a positive impact on the owners, contributing to the sustainability of the community and its
surroundings.
Key words: function, family garden, aesthetic.
INTRODUCTION aesthetic of the garden. An intercropping

system besides its ornamental values has a
The first vegetable gardens were situated on the positive impact in attracting useful insects.
valleys of Tigre, Eufrat and Indus rivers Using this system in a family vegetable garden
(Iliescu, 2008), initially their purpose was to can have many advantages that can have a good
offer fresh vegetables and, in some celebration, impact on the community and on the
they had an ornamental use (Sima, 2009). environment.
Landscape design is a phenomenon that was In this context, the paper presents an analysis of
used in vegetable growing for a long time, due the effects of an ornamental vegetable garden
to the desire to create an area where you can on the sustainability of the family and
relax and admire beautiful views. This community in which it exists, in the period
landscape offers food for the body and soul 2014-2017.
(Galea et al., 2017).
Father Ignatius said „a garden should look like MATERIALS AND METHODS
a beautiful arranged table”. In a vegetable
garden, the soul of the owner is represented by In order to reach the aim and the objectives of
an element in the design, usually a flower this research six experimental gardens were
(Sima, 2017). conceived and based on the analyses obtained
In public spaces, in Romania, ornamental in different studies (Galea et al., 2017) one
vegetable garden are rare, but because of the garden was chose as the best from an
importance that vegetable crops have and the ornamental and sustainable point of view.
desire for healthy vegetables in private gardens, The experimental garden was done/ established
this type of design has slowly entered in the experimental field of the vegetable
Romanian households (Galea, 2017). growing department from USAMV Iasi,
Vegetable plants have many ornamental between 2014-2017.
elements that in an intercropping system are The studied family ornamental vegetable
underlined and contribute to the general garden was designed in a geometrical style with
517
a planted area of 100 m2. The family garden values in line with those in known literature,
was evaluated by a panel of experts in a survey fact that proves that the applied intercropping
regarding its ornamental value and in another, system has an evident effect in obtaining a
regarding its sustainability value. The two qualitative production (Table 2).
surveys had scale from 1 to 5, in wich 1 The percentage of sucrose for cherry tomatoes
represented highley disagree, 2 represented varied from 6.09% for Aristan purple bumble
disagree, 3 neutral, 4 agree and 5 represented bee cv. to 8.15% for Ema de Buzau cv.
higly agree. Regarding the hot pepper the obtained result
In the composition of the surveys were taken were higher than the ones in the known
into consideration the following: the combining literature with an average value of 8.35%.
method, the proposed species, the ornamental The obtained result for egg plants was 3.6-
layout, garden functions (educational, cultural 3.8%, values that are in between the known
etc.), the influence of the used intercropping data (2.5-4.0%) (Bodea and Enăchescu, 1984).
system on plant growth, development and
production, pest and disease attack. Table 1. Comparative results regarding the obtained
The achieved results were evaluated using production
Nr. Plant Standard
SWOT analysis to help determine the dree of Crt.
Plant name
production (g) production (g)
ornamental and agronomical value. 1. Lollo Rossa lettuce 180 - 200 200-300
From an economic point of view, plant produc- 2. Aristan Puple Bumble 1380 1500
Bee tomatoe
tion and production quality was assessed. 3. Beam’s yellow pear 1200 1500
Due to the fact that the vegetable garden has in tomatoe
4. Ema de Buzău tomatoe 1400 1500
its composition 34 different vegetable plants 5. Ovari feher squash 1700 430-450
(13 intercropping systems), 6 samples 7. Bordi pea 3 3
(tomatoes, hot pepper, eggplant and squash) 8. Runner bean 80 70
9. Thyme 200 125-175
were analysed regarding production quality 10. Pascal celery 300-342 120
using the device DR 301-95 (digital 11. Black beauty eggplant 3000 900-1800
12. Kayene pepper 280 63-105
refractometer-for sucrose) and a vacuum drying 13. Sweet Thing F1 sweet 775 750
oven (for dry substance and humidity). corn
17. Scarlet kale 522 792
In order to realize the purpose of this study, 18. Kadet kale 540 792
besides increasing the ornamental value, one 19. Chard 3500 41,66
the main objectives was to increase the impact 20. Dalmatziano beans 40 20
21. Clemson Spineless okra 210 105
of the vegetable garden on its owners and on 22. Cap de Negru 2 cabbage 2315 1400-1800
the environment. 23. Calabrese Natalino 450 500
broccoli
24. De Ciorani hysopus 200 193
RESULTS AND DISCUSSIONS 25. Violet basil 190 105-126
26. Onions 26 26
27. Piccolo verde di Parigi 800 1050
The climatic conditions from the experimental cucumber
field were suitable for vegetable growing. From 28.
30.
Sage
Di Sicilia Violetto
550
450
360
480-800
a development point of view the habitus of the cauliflower
plants was harmonious. 31. Bulgăre de zăpadă 600 480-800
cauliflower
The chosen intercropping systems had a 32. Radish 425 200-300
positive impact on plant production (Table 1). 33. Pumpkin butternut 10500 10000-12000
34. Rosa di trevise 4 cicory 220 177-222
The average values obtained for the studied
plants were close to the values found in the
Regarding dry matter the values obtained for
existing literature (Stan et al., 2003; Stan and
cherry tomatoes were between 8.09-13.20%.
Munteanu, 2001; Fălticeanu and Munteanu,
The highest values was for Ema de Buzau cv.
2003; Savițchi et al., 2013; Multeanu, 2003;
The average values obtained for eggplant were
Ciofu et al., 2003; Carvalho et al., 2012;
6.8% for dry matter for Black beauty cultivar
Tringovska et al., 2015; Tincă, 2011;
and 7.2% for Albe de Buzău cv.
Hamburdă, 2015; Bavec et al., 2010). The
The obtained results for squash had an average
results optained regarding chemical
of 5.4% for dry matter.
composition, of the six analyzed samples had
518
Based on the two surveys the strong points and had a good influence in plant production
the weaknesses of the garden were underlined (Figure 2).
using SWOT analysis providing an overview of
the ornamental degree of the garden and over
its effects regarding the sustainability of the
community and its surroundings.
The obtained results regarding the ornamental
value of the studies garden revealed that the
ornamental vegetable garden has many
functions, with beautiful perspectives. The
usage of different intercropping systems
(lettuce + runner bean; eggplant + pepper; okra
+ beans etc.) contributed in a positive way to Figure 2. Evaluation forms analysis for garden
the ornamental degree (Figure 1). sustainability degree
Based on the SWOT analysis the garden has

many strengths, such as the education of the
young generation, high yield, high ornamental
value, the increase of useful insects and fresh
vegetable for a long period.
The analysis also revealed that the garden
needs many carrying practices. By using an
intercropping system, the number of disease
attack was reduced.
From an economic point of view the obtained
results reveal that the highest cost in the
ornamental garden were the ones with
materials, fallowed by seeds and seedlings cost.
Figure 1. Garden detail
In the first year of study the profit rate was 11
One of the week point of the garden was % due to the initial investments (Table 3). The
represented by the alternative mean of plant profit rate was obtained din the second year of
protection, that need to be improved (Table 2). study.
Table 2. SWOT Analysis - Model 6 Table 3 Correlation study between costs and revenue
Strengths Weaknesses Nr.crt. Specification U.M. 1st 2sd 3rd
• many functions; • alternative mean of plant year year year
• beautiful perspectives; protection; 1. Total production lei 1759 1028 1229
• the style of the garden costs
enhances the ambiance of 2. Yeald value lei 1856 1663 1600
the household; 3. Profit lei 204 635 370
• the intercropping system 4. Profit rate % 11.65 61.77 30.08
brings aesthetic value;
Opportunities Threats
• landscape design principles CONCLUSIONS
correctly used ; • reduced communication.
• color and volume game -
increased ornamental The proposed intercropping systems had a
impact positive effect on plant growth and develop-
ment. The ornamental perspectives created in
The surveys regarding the sustainability of the the garden were beautiful and contributed to the
ornamental family vegetable garden revealed enhancement of the environment. The many
that according to the panel of experts the function the garden has (ornamental, aesthetic,
garden is considered economically affordable. cultural and educational) increase community
The studied intercropping systems had a communication and help the owners maintain a
positive impact on attracting useful insects that healthy life style.
519
From an economic point of view the condițiile județului Iași, Teză de doctorat, USAMV
ornamental vegetable garden contributes to the Iași, 100-215.
Fălticeanu M., Munteanu N., 2006. Plante utile pentru
economy of the family budget. grădina dumneavoastră. Editura Tipo Moldova, Iași,
By using an intercropping system, the garden 35-115.
offers fresh vegetable for a long time. Galea (Deleanu) Florina-Maria, Munteanu N., Stoleru
V., Teliban G.C., Gache (Lungu) Mirabela, Hriscu
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Pelaghia, Apahidean S., Horgoş A., Berar V., Lauer Stan N.T, Munteanu N., Stan T.N., 2003. Legumicultură,
K.F., Atanasiu N., 2003. Tratat de legumicultură. vol. III. Editura “Ion Ionescu de la Brad”, Iaşi, 20 -
Editura Ceres, Bucureşti 100-200. 156 .
Costanda Gabriela (Tincă), 2011. Cercetări privind
elaborarea tehnologiilor ecologice de cultivare a unor
specii legumicole condimentare și aromatice în
520
Floriculture,
Ornamental Plants,
Design and
Landscape
Architecture
521
522
PRELIMINARY RESEARCH ON THE INFLUENCE OF THE PLANTING

TIME ON THE GROWING AND BLOOMING OF VARIETY OF
HERBACEOUS PEONY IN THE FIELD
George Nicolae CAZAN, Sorina PETRA, Florin TOMA

Abstract
The present paper refers to the results of the research on the influence of the different planting times of some varieties
of herbaceous peony in the field on the blooming and growing. The research was performed in the experimental field of
USAMV Bucharest and in the town of Singureni, Giurgiu County. The studies made are based on the data obtained both
by visual observation and by biometric measurements on the varieties of herbaceous peony. The biologic material war
represented by a variety of herbaceous peony with white flowers, with pink flowers and with red flowers. The data was
processed in the following indicators: growth stages (emergence, springing, percentage of shoots starting vegetation,
percentage of springs, vegetation rest) and blooming stages (percentage of flower springs, appearance of buds, stages
of blooming, percentage of blooming) as well as the dynamics of growth and blooming. There were results related both
to the growth and blooming stages and to the dynamics of the growth and blooming.
Key words: blooming, growth, herbaceous peony, planning times.
INTRODUCTION Paeonia peregrina Miller and Paeonia mollis

Hort., as well as the herbaceous hybrids. The
The peony is native of Europe, Asia and species Paeonia lactiflora Pallas and its
America. The peony grows spontaneously in growing are the last to come at rest on the
numerous countries with temperate climate in ecologic conditions of the area, as well as the
the Northern hemisphere. Peonies are perennial Itoh hybrids.
ornamental plants of the genus Paeonia, At the same time, the crops of the species
pertaining to the family of Paeoniaceae and it Paeonia lactiflora Pallas became remarkable
consists of 33 species (Rogers, 1996). by the decorative colour of the foliar apparatus,
The herbaceous peonies are grown mainly for especially ‘Rosy Down’ and ‘Auguste Dessert’.
the use in horticultural industry like a home In a research study published in 2002 by
garden plant and also, the peonies are Barzilay et al., with the topic The annual life
commercially grown as cut flowers. Peonies cycle and flower development of ‘Sarah
are, also cultivated for their medicinal Bernhardt’ under the conditions of Israel, they
properties (Dong-Yi et al., 2011) and even for present the life cycle and the morphogenesis of
their use as ornamental plants (Nehrlingand, the floral sprouts of Paeonia lactiflora Pallas.
1978). The cultivar ‘Sarah Bernhardt’ was studied on
The research made by Cucu et al.(2009) on the the conditions of Israeli climate.
study of the behaviour in crop of some species The formation of the buds starts at the
and the growing of the peony in the southern beginning of spring. The stems grow rapidly
part of Romania, under the influence of local and they reach heights of 50-70 cm in 60-70
climate factors. days. The flowering starts in April and it
The study started with the plants coming at rest continues until the end of May. After the
in the autumn of 2005. It was observed that the flowering, the stem remains green until
first signs of senescence are given away by the September-October, when the leaves age and
species Paeonia officinalis L. and its growing the plant comes at vegetation rest for 3-4
(late August), followed closely by the species months.
523
Toma (2009) mentions that the herbaceous Table 1. Experimental variants
species of peony reproduce by the division of Number
Diameter
the bush, operation made by the mature plants, Date of Type of
of
of the
ballot of
Variant Cultivar sprouts
at the age of 4-5 years. After part of the soil is planting material
min/
the roots
min/max
removed from the roots, the mother plants are max
(cm)
cut by hand or with a very sharp knife in 3-6 Cultivar

with pink
Divided
parts, depending on the size of the mother V1 flowers in the
collectionUS
05.11.2016
bushes
1-7 2 - 16
plant. AMVB
After the division, the very large leaves are Cultivar

with pink
Divided
shortened to 2-3 foils in order to assure a good V2 flowers in the
collectionUS
14.11.2016
bushes
2 - 16 5 - 21
reproduction of the plants. The plant divisions AMVB

Cultivar
will be put in planting holes, after mudding the with pink Divided
V3 05.11.2016 4-9 8 - 15
roots, so that the area of the collar is covered flowers in
SINGURENI
bushes
with 2-3 cm of soil (Cantor, 2016). Deeper Cultivar

with pink Undivided
planting results in the delay and the V4
flowers
14.11.2016
bushes
8 - 24 20 - 27
diminishing of the flower or even its SINGURENI

Cultivar
compromise for the next 2-3 years from the V5
with pink
25.11.2016
Divided
2 - 14 6 - 17
flowers in bushes
planting (Toma, 2009). SINGURENI
Peonies need a long period of cold of at least Cultivar
with pink Undivided
900 hours around or under the freezing point V6
flowers in
25.11.2016
bushes
9 - 16 22 - 27
SINGURENI
(Jacob et al., 2006), in order to pass through Cultivar
the rest period and start the vegetation in the V7
with white
flowers in
05.11.2016
Divided
bushes
4-9 9 - 15
next season. In case the temperature does not SINGURENI
drop enough, the peonies will fail to make Cultivar
with white Divided
V8 14.11.2016 3 - 12 5 - 14
flowers. flowers
SINGURENI
bushes
Cultivar
MATERIALS AND METHODS V9
with red
flowers in the 05.11.2016
Divided
5 - 13 5 - 16
bushes
collection
USAMVB
The planting material used in the research was
constituted of herbaceous peony with white
flowers, with red flowers and with pink
flowers. The varieties of peonies were planted
both at USAMV Bucharest and in Singureni in
3 planting times, this resulting 9 variants.
The planting was made with plants coming
from 3-4 years old bushes and 7 years old
bushes as divided and undivided bushes
(Figures 1-4). The planting occurred in the
autumn of 2016 at USAMV Bucharest and at
Singureni. The data was processed using the
biometric measurements and the visual Figure 1. Undivided bushes, pink cultivar in Singureni
observations made during 2017.
The indicators used in the study of the research
were: start of vegetation, the appearance of
shoots, the senescence, the appearance of the
sprouts, the percentage of shoots, the percen-
tage of flowering shoots, the phenophases of
growth and flowering, the dynamics of growth
and flowering, the earliness of the flowers and
the flowering percentage.
The experimental variants, the cultivar the
plating dates and the features of the planting
materials are given in Table 1. Figure 2. Divided bushes, white cultivar in Singureni
524
presented the growth of the shoots with the late
start of sprouts.
The growth of shoots in the two research
locations of the varieties of herbaceous peony
studies is presented in Figures 10-13.
Growth stages of the varieties of herbaceous peony
20.mai
30.apr.
10.apr.
21.mar.
01.mar.
09.feb.
20.ian.
V1 V2 V3 V4 V5 V6 V7 V8 V9
Figure 3. Divided bushes, pink cultivar in Singureni appearance of buds 10.mar14.mar05.mar12.mar18.mar28.feb01.mar27.feb13.mar
end of appearance 31.mar27.mar07.apr 07.apr 07.apr23.mar24.mar21.mar03.apr
growth of shoots 11.mar15.mar06.mar13.mar29.mar01.mar02.mar28.feb14.mar
end of growth 01.mai04.mai28.apr04.mai07.mai22.apr 25.apr 20.apr 28.apr
Figure 5. Stages of growth of the varieties of herbaceous

peony planted in the field in 2017
Figure 4. Divided bushes, red and pink cultivar in

USAMV Bucharest
The obtained results as far as the phenophases

of growth of the varieties of herbaceous peony
studied were made by the measurement of the Figure 6. Appearance of sprouts
in the herbaceous peony, 2017
following indicators: date of appearance of the
sprouts, end or appearance of the sprouts’,
appearance of the shoots and the end of the
growth of the shoots.
When starting the vegetation, the following
results were obtained: early start in the variants
V6, V7 and V8 and late starts in the variants
V2, V4, V5 and V9 according to figure 5. In
the early appearance of the shoots, the variants
V6, V7 and V8 stood out, and in the late
appearance, the variants V2, V4, V5 and V9.
In her research Barzilay et al. (2002) at the
‘Sarah Bernhardt’ paeony cultivar, the shows
that on the conditions of Israeli climate the Figure 7. Start of vegetation in the varieties of
formation of the buds starts at the beginning of herbaceous peony at USAMV Bucharest, 2017
spring.
In Figures 6 and 7 it is observed the appearance
of the sprouts in the two locations of the
research on 2017, and in Figures 8 and 9 it is
525
Figure 8. Start of vegetation and the growth of shoots, Figure 11. Growth of shoots, white and pink cultivar of
pink cultivar of Singureni, 2017 Singureni, 2017
Figure 9. Appearance of the sprouts and of the shoots in Figure 12. Aspect of e bush and shoots, pink cultivar of
white cultivar of Singureni, 2017 Singureni, undivided bushes, 2017
Figure 13. Aspect of the shoots towards the end of

Figure 10. Growth of shoots in the varieties of growth, pink cultivar of Singureni, 2017
herbaceous peony planted in USAMV Bucharest, 2017
526
Researching the stages of growth and
flowering, the following indicators were
analysed: percentage of vegetation start,
percentage of shoots, percentage of flower
shoots, percentage of plants with floral shoots,
percentage of plants with blooming floral
shoots and the blooming percentage.
The percentage of vegetation start in all the
variants is 50%, the same with the percentage
of shoots. The percentage of flower shoots was
in two variants under 50%, and the rest above
50%. As far as the percentage of plants with
floral shoots, the obtained results being under
100% in two variants and 100% in the others.
The blooming percentage was under 50% in Figure 15. Aspect of flowering in pink cultivar at
variants V1, V2, V4, V5, V6, V7, and above USAMV Bucharest, May 24, 2017
50% in variants V3, V8 and V9. The results are
given in Figure 14.
Perecentage in the stages of growth and flowering of the

varieties of herbaceous peony
120,00%
100,00%
80,00%
60,00%
40,00%
20,00%
0,00%
V1 V2 V3 V4 V5 V6 V7 V8 V9
percentage of vegetation start 94,00% 70,30% 71,21% 89,07% 73,61% 89,93% 66,66% 92,38% 92,59%
percentage of shoots 94,00% 70,30% 71,21% 89,07% 73,61% 89,93% 66,66% 92,38% 92,59%
percentage of flower shoots 29,78% 22,85% 53,19% 68,86% 54,71% 88,05% 60,00% 72,16% 68,00%
percentage of plants with floral
54,54% 71,42% 100% 100% 100% 100% 100% 100% 100%
shoots
percentage of plants with
Figure 16. Aspect of flowering in white cultivar
blooming floral shoots
33,33% 20,00% 72,72% 100% 63,63%100,00%42,85% 75% 28,57%
Singureni, May 24, 2017
blooming percentage 21,42% 12,50% 50% 42,66% 44,82% 83,05% 22,22% 54,28% 13,88%
Figure 14. Percentage of growth and flowering in the

varieties of herbaceous peony planted in the field, 2017
The aspect of the mass flowering of the

varieties of herbaceous peony is presented in
Figures 15 and 16. In Figures 17 and 18 are
given the plants of herbaceous peony with
floral shoots. Figure 19 presents the aspect of
the flower in the cultivar of peony with pink
flowers in USAMV Bucharest.
Figure 17. Floral shoots in pink cultivar
in Singureni, May 28, 2017
527
Number of shoots of the varieties of herbaceous peony
120
100
80
60
40
20
0
V1 V2 V3 V4 V5 V6 V7 V8 V9
Total shoots 47 35 46 106 53 67 30 97 50
Total floral shoot s 14 8 25 73 29 59 18 70 34
Figure 18. Floral shoots in pink cultivar at USAMV Total flowered floral
3 1 12 31 13 47 4 32 4
Bucharest, May 15, 2017 shoots
Figure 20. Number and types of shoots of the varieties of

herbaceous peony planted in the field, 2017
Figure 19. Aspect of flowering in pink cultivar USAMV

Bucharest, May 24, 2017
We went on to analyse also the number of Figure 21. Start of flowering in the white cultivar
shoots in the varieties of herbaceous peony in Singureni, 16.05.2017
studied in the indicators: total shoots, total
floral shoots and total flowering floral shoots.
The number of shoots in the variants of divided
bushes was a minimum of 35 in V2 and a
maximum 97 in V8, and in the variants with
undivided bushes they were 67 in V6 and 106
in V4.
The minimum number of floral shoots was in
V2 and the maximum in V8 and V4. The
minimum number of flowering floral shoots
was in V2, and the maximum in V6. The
obtained results are given in figure 20.
Figure 22. Floral shoots in the red cultivar
Figures 21 and 22 present shoots of peony
USAMV Bucharest, May 10, 2017
before the flowering with semi-open buds in
the two research locations. The early emergence of the bud appeared in
variants V6, V7 and V8 and the late emergence
in variants V2, V3, V5, and V9.
The closed bud phase May 4 in V6, May 10 in
V8, May 12 in V3, May 13 in V5,V7 and V9,
May 15 in V1,V2,V4.
The opening of the flower emerged in May 10
in V6, May 15 in V8, May 17 in V3, V5; May
528
18 in V7 and V9; May 19 in V1 and May 20 in
V2.
The end of blooming occurred on May 25 in
V8; May 27 in V6; May 30 in V5; June 2 in V3
and V7; June 4 in V2; June 5 in V4 and V9;
June 6 in V1.
Stages of flowering in the varieties of herbaceous peony
06.iun.04.iun.02.iun.05.iun.30.mai27.mai02.iun.25.mai05.iun.
22.mai23.mai19.mai23.mai20.mai12.mai20.mai18.mai21.mai Figure 25. Red cultivar Figure 26. Pink cultivar

19.mai20.mai17.mai21.mai17.mai10.mai18.mai15.mai18.mai
flowered in USAMV flowered in USAMV
Bucharest, May 2017 Bucharest, May 2017
27.mar.09.apr.10.apr.04.apr.10.apr.18.mar.
20.mar.
20.mar.09.apr.
V1 V2 V3 V4 V5 V6 V7 V8 V9
end of flowering 06.iun.04.iun.02.iun.05.iun.30.mai27.mai02.iun.25.mai05.iun.
tota opening of flower 22.mai23.mai19.mai23.mai20.mai12.mai20.mai18.mai21.mai
opening of flower 19.mai20.mai17.mai21.mai17.mai10.mai18.mai15.mai18.mai
semi-open coloured bud 17.mai17.mai14.mai18.mai15.mai08.mai15.mai13.mai15.mai
closed bud 15.mai15.mai12.mai15.mai13.mai04.mai13.mai10.mai13.mai
emergence of bud 27.mar.09.apr.10.apr.04.apr.10.apr.18.mar. 20.mar.09.apr.
20.mar.
Figure 23. Stages of flowering in the varieties of

herbaceous peony planted in the field, 2017 Figure 27. Aspect of Figure 28. Aspect of
flowering in the red flowering in the pink
cultivar USAMV cultivar USAMV
Figures 24-28 present the aspects of flowering Bucharest, May 24, 2017 Bucharest, May 24, 2017
and flowers of varieties of herbaceous peony in
2017. Analysing the duration of the stages of growth
and flowering of the varieties of herbaceous
peony, it was observed that the shortest
duration of budding was in variants V2, V5 and
V9, and the longest in variant V3. The duration
of flowering was of 13 days in V5 and V8, 14
days in V4 and V7, 15days in V2, V3, V6, and
17 days in V1 and V9 (Figure 29).
Duration of stages of growth and flowering in the varieties of

herbaceous peony (days)
60
50
40
30
Figure 24. Aspect of flowering in the red cultivar 20
USAMV Bucharest, May 24, 2017 10
0
V1 V2 V3 V4 V5 V6 V7 V8 V9
duration of total budding 21 13 32 25 19 25 23 24 18
duration of shoot growth 48 47 51 48 54 52 52 50 42
duration of flowering 17 15 15 14 13 15 14 13 17
duration of plant flowers 14 13 13 12 11 10 11 8 16
Figure 29. The duration of stages of growth and

flowering in the varieties of herbaceous peony planted in
the field, 2017
Figures 30-38 present the aspect of flowering
and the aspect of the flower in the cultivars of
herbaceous peony with pink flowers in
529
USAMV Bucharest, with pink flowers in
Singureni and with white flowers in Singureni.
Figure 33. Aspect of flower, pink cultivar Singureni,

May 18, 2017
Figure 30. Aspect of flower, pink cultivar USAMV
Bucharest, May 24, 2017
Figure 31. Aspect of flower, pink cultivar USAMV Figure 34. Aspect of flower, white cultivar Singureni,
Bucharest, May 24, 2017 May 20, 2017
Figure 35. Aspect of flower, white cultivar Singureni,

Figure 32. Aspect of flower, pink cultivar Singureni, May 20, 2017
May 24, 2017
530
Figure 36. Aspect of flower, white cultivar Singureni,
May 22, 2017
Figure 38. Total opening of the flowers and the
aspect of the flowering in the pink cultivar
in Singureni, May 24, 2017
CONCLUSIONS
The research made shows that some varieties of

herbaceous peony may be planted in a period
different from August or September, increasing
thus the planting period in autumn of the
herbaceous peony. This research came to the
Figure 37. Aspect of flower, pink cultivar Singureni, following conclusions:
May 20, 2017 The percentage of the start of vegetation in all
the variants was over 50%, respectively
As far as the dynamics of the growth and of the 66.66% in the cultivar with white flowers in
inflorescence of the types of studies herbaceous Singureni by 94% in the cultivar with pink
peony, the indicators in Table 2 were analysed. flowers in the collection USAMV Bucharest.
The short vegetation was observed for the The percentage of shoots is the same with the
variants V1, V6, V7 and V9, and a longer percentage of start of vegetation.
vegetation period was observed in variants V2, The percentage of floral shoots in two variants
V3, V4, V5 and V8. was under 50% respectively in the cultivar with
pink flowers in the collection of USAMV
Table 2. The dynamics of growth and flowering in the
varieties of herbaceous peony planted in the field, 2017 Bucharest cu 29.78% and 22.85%, and the rest
Variant Sprouting Shooting Budding Blooming End of End of
of the variants is over 50%, starting with the
blooming vegetation variety with pink flowers in Singureni with
V1 53.19% by 88.05% in the cultivar with red
10.03 13.03 27.03 19.05 06.06 III
October flowers in the collection of UASVM Bucharest.
V2
01.03 03.03 20.03 18.05 02.06
I
November The percentage of plants with floral shoots is
V3
05.03 07.03 10.04 17.05 02.06
I
November over 50% in all the variants, of which two
V4
12.03 16.03 04.04 21.05 05.06
I
November variants with 54.54% and 71.42% in the
V5
18.03 21.03 10.04 17.05 30.05
I
November
cultivar with pink flowers in the collection of
V6
14.03 17.03 09.04 20.05 04.06 III
USAMV Bucharest and the rest of 7 variants of
V7
October 100% respectively in the cultivars with white
13.03 16.03 09.04 18.05 05.06 III
October
flowers in Singureni, with red flowers in the
V8 I collection of USAMV Bucharest and with pink
27.02 01.03 20.03 15.05 25.05 November
V9 flowers in Singureni.
28.02 01.03 18.03 10.05 27.05 III
October
531
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Medicine. Frontiers in Pharmacology, vol. 2, Front.
Barzilay A., Zemah H., Kamenetsky R., Ran I., 2002. Pharmacol., 25 February 2011 |
The annual life cycle and flower development of https://doi.org/10.3389/fphar.2011.00010.
‘Sarah Bernhardt’ under the conditions of Israel. Hort Jacob Y., Mastrantuono S., Ferrero F., 2006. Improving
Science, vol. 37 (2), 300-303. the efficient of herbaceous peony breeding methods.
Cantor M., 2016. Special Floriculture. AcademicPres ISHS Acta Horticulture, vol. 714: 59-66.
Publishing House, Cluj Napoca, 96-98. Nehrling A., Nehrling I., 1978. Peonies, Outdoors and In.
Cucu E.I., Şelaru E., 2009. Preliminary results regarding 2nd ed. New York: Hearthside Press, 2-23.
the behaviour in culture of some peony cultivars. Rogers A., 1996, Peonies. Timber Press, 384 pages.
Scientific Papers of USAMV Bucharest, Series B, Toma F., 2009. Floriculture and Floral Art, Vol. IV.
Horticulture, vol. LIII, 224-228. Grown species for parks and gardens decoration.
Dong-Yi He, Sheng-Ming Dai, 2011. Antiinflammatory Invel Multimedia Bucharest Publishing House, 228-
and Immuno-modulatory Effects of Paeonia 232.
532
PRELIMINARY RESEARCH ON THE GROWTH AND FLOWERING
OF DIFFERENT VARIETIES OF HERBACEOUS PEONY UNDER CONDITIONS
OF CONTAINERIZED AND FORCED CULTIVATION
George Nicolae CAZAN, Sorina PETRA, Florin TOMA

59 Mărăști Blvd, District 1, Bucharest, Romania
Abstract
This research has been focused on the studies on the growth and flowering of some varieties of herbaceous peony under
conditions of containerized and forced cultivation. The research was conducted at USAMV Bucharest using as biologic
material split bushes with white, pink and red flowers originated from USAMV Bucharest and Singureni, Giurgiu
County. The split bushes being planted in pots of 30cm in diameter in autumn in October. After planting, the pots were
kept in cold conditions a different number of weeks, after which they have been brought in the greenhouse to be forced.
For each of the forced stages, observations have been made, regarding the following phenophases of growing and
developing: starting in vegetation, number of sprouts, growth of the sprouts, appearance of the buds, opening of the
flowers, fading of the flowers, the flowering time, the flowering percentage. The results obtained show a good
behaviour of the herbaceous peony in containerized and forced cultivation, the percentage and the quality of blooming
being variable, depending on the variety and the moment of the introduction of the pots in the greenhouse.
Key words: containerized and forced culture, early flowering, herbaceous peony.
INTRODUCTION they bloom in May-July. Anyway, there are

some researches that prove the capacity of the
Peonies are among the most popular garden herbaceous peony to bloom under warmer
plants in temperate regions. Herbaceous climatic conditions, e.g. California and Israel
peonies are also widely sold as cut flowers, (Byrne et al., 1986; Halevy et al., 1995).
although they are generally available only at Other researches prove that peonies can be
the end of spring and early summer. The annual forced to bloom in greenhouse conditions
growth is predetermined: if the growing tip of a (Wilkins et al., 1985; Evans et al., 1990) for the
sprout is removed, no new buds will develop in early production of cut flowers and potted
that season (Kamenetsky et al., 2003, 2012). plants.
In temperate regions of the Northern Gregory et al. (2015) elaborated a database for
hemisphere, the peonies are blooming in May - herbaceous peony cultivated in warm climate
July. Modern reproduction researches from the zones, in the "Effects of temperature on
last years lead to obtain of cultivars with stagnation and plant growth" article.
flowers with very different shapes and colours These authors introduce the first database for
(Cantor, 2016). development parameters for the herbaceous
Some of these new cultivars were studied also peony cultivated in a warm areas. The data on
in the climatic areas of the Southern Romania the temperature effect during the rest and
(Cucu et al., 2009). growth period are presented for 2,232 plants of
In the Northern hemisphere there is a growing two commercial varieties exposed to 14
demand to extend the flowering period of the different temperature regimes.
peony (Toma, 2009). For example, Alaska Recently, the possibility of cultivating peony in
peonies are nowadays foremost only for export warm climates has been reported also by the
(Fitzgerald, 2004). Another strategy is to use authors Fulton et al. (2001) and Catley et al.
cold artificial environments as well as (2001).
hormonal treatments to induce flowering at Evans et al. (1990) and Wilkins et al. (1985)
different times of the year (Halevy et al., 2002). showed that in milder areas the peonies may be
Herbaceous peonies are grown successfully in forced to produce early cut flowers in
moderate climates with cold winters, where greenhouse.
533
In order to increase early forcing and to ones in open field (9°C average daily
improve the profitability of growing peonies in temperature and 2°C the average nightly
warmer conditions, in-depth knowledge of temperature). Here, they have been kept for 10
temperature requirements is required at weeks, until the frost.
different stages of the annual growth cycle From this moment, the pots were moved in a
Gregory et al. (2015). basement with an average temperature of 5°C,
In a previous study, Gregory et al. (2015) where they have been kept a variable number
established the key relationships between the of weeks, until the movement in the greenhouse
thermal regime in the rest period, the stem for forcing period.
growing and blooming. The movement of the plants pots in the
The cooling regime for the release from resting, greenhouse for forcing was made at four
was studied on peony plants grown in the field different dates: 14th of February, 21st of
by exposing them to a cold winter climate in February, 28th of February and 7th of March
northern Israel (8-10°C by night, 16-26°C by 2017. By combining these dates with the
day, average about 17°C from November to varieties of colours and provenance, resulted 11
February). experimental varieties (Table 1).
The greenhouses were covered with plastic
films on different dates, looking for the Table 1. Experimental Variants
accumulation of cooling units (Fishman et al., Variant Cultivar No. of cold Date of Number Diameter of
weeks forcing of buds roots bale
1987; Erez et al., 1988). The flowering min - max min – max
(cm)
occurred two months after the resting of the V1 White flower
16
plants has been over (Halevy et al., 2002). peony from
SINGURENI
14.02.2017 2-9 3 - 10
The peonies cultivated in the pots have been V2 White flower

peony from 17 21.02.2017 5-7 3 - 10
artificially cooled by refrigeration to 2°C for 60 V3
SINGURENI
White flower
days (‘Sarah Bernhard’, 14) or 1-7°C for 3-12 peony from
SINGURENI
18 28.02.2017 5 8-9
weeks (‘Coral Sunset’, ‘Monsieur Jules Elie’, V4 Pink flower

peony from 16 14.02.2017 3 - 10 4 - 13
‘Sarah Bernhardt’) (Barzilay et al., 2002). V5
SINGURENI
Pink flower
The time until the start of vegetation after the peony from
SINGURENI
17 21.02.2017 3 - 11 4 - 12
completion of the cooling treatment has V6 Pink flower

peony from 14.02.2017 4 6-9
decreased proportionally with the increase in the
USAMVB
16
the duration of the cooling treatment (Hall et collection

V7 Pink flower
al., 2001). peony from 21.02.2017 1-7 2-4
the 17
In Chinese varieties, stem height, flower size USAMVB
collection
and flowering rate were also positively affected V8 Red flower
by the cooling temperature during the resting peony from

the 16
14.02.2017 3-8 4 - 15
period (Cheng et al., 2009). USAMVB

collection
V9 Red flower
peony from 21.02.2017 2 - 10 4 - 11
MATHERIALS AND METHODS the
USAMVB
17
collection
V10 Red flower
The biological material used in these researches peony from
the 18
28.02.2017 3 - 12 5-9
is made up of divided herbaceous peony USAMVB

collection
varieties with white, pink and red flowers. V11 Red flower
peony from 07.03.2017 2-6 2-4
These were obtained from mother plants aged the
USAMVB
19
6-7 years, from USAMV Bucharest and collection
Singureni, Giurgiu County.

The plant divisions were planted in October, in The observations on plants had in view the
pots with a diameter of 30 cm. The subsoil of following elements: starting of vegetation,
planting was consisting of 40% peat, 30% soil, number of sprouts, the growing of sprouts, the
20% soil naturally fertilised, 5% sand and 5% apparition of flower buds, the number of shoots
perlite with a granulation of 4 mm. with flowers, the opening of flowers, the fading
After planting, the pots were left outside in a of flowers. Based on these observations, the
shaded area in conditions corresponding to the duration of blooming and the flowering
percentage were calculated. Also, the possible
534
correlations between the plant growing
elements of the plants and between them and
the duration of the cold period administrated
before the introduction of the plants in the
greenhouse.
The starting on vegetation was faster on the

variants V6 and V8 on 09.02., besides V1 on
10.02., V4 on 11.02., V2,V5, V9 on 15.02., V3
and V7 on 17.02., V10 on 19.02. and V11 on
28.02. (Figure 1).
The appearance of the sprouts was observed
starting with 10.02. at V6 and V8 until 20.02. Figure 2. Growth of Figure 3. Growth of
at V10 and 01.03. at V11 (Figure 1). sprouts, red cultivar, sprouts, red cultivar,
UASVM Bucharest, UASVM Bucharest,
V10, 2017 V8, 2017
Figure 1. The phenophases of herbaceous peony

growing, forced in pots, 2017
Figure 4. Starting of Figure 5. Growing aspect
vegetation of the pink of sprouts of pink cultivar
We are concluding that the varieties with pink cultivar from UASVM from Singureni, 2017
and red flowers from USAMV Bucharest are Bucharest, V6, 2017
starting earlier in vegetation compared to the
white and pink flowers varieties from
Singureni. Also, a bigger number of weeks of
cold between the introduction of the pots in the
greenhouse lead to a quicker vegetation starting
on all varieties.
Details regarding the growth phenophases are
shown in Figures 2-6.
Duration of buds starting was between 6 days
at V7 and 21 days at V9, and the sprouts
growth lasted between 16 days at V10 and 38
days at V9 (Figure 7).
The growing of sprouts lasted from 16 days at
V11 up to 38 days at V9 (Figure 7).
Figure 6. Growing and trellising on
peony sprouts, 2017
535
Table 2: Growing dynamics of the herbaceous peony
The duration of phenophises at red flowers peony variety from
U.S.A.M.V. Bucharest collection
varieties planted in pots and forced, 2017
38 Variant Cultivar Buds % Start of
40
36 appearance vegetation sprouts
34 start growth
35 White flowers February 10 33.33% February 11
V1 peony from
30 28 SINGURENI
White flowers February 15 36.36% February 16
V2 peony from
25
SINGURENI
days
21
White flowers February 17 90% February 18
20 V3 peony from
16 16 SINGURENI
15 Pink flowers February 11 64% February 12
11 V4 peony from
10
SINGURENI
10
6 Pink flowers February 15 86% February 16
V5 peony from
5 SINGURENI
Pink flowers February 9 100% February 10
0 V6 peony from
V7 V8 V9 V10 V11 USAMVB
the total duration of buds collection
6 11 21 16 10 Pink flowers February 7 95% February 18
starting
V7 peony from
the duration of sprouts USAMVB
34 36 38 28 16
growing collection
Red flowers February 9 63.61% February 10
V8 peony from
Figure 7. Dynamics of flowering, red flowers peony USAMVB
collection
cultivar, UASVM Bucharest collection, 2017 Red flowers February 15 63.12% February 16
V9 peony from
USAMVB
collection
It is observed a shorter period from total Red flowers February 19 91.90% February 20
starting of the buds at the pink flowers variety V10 peony from
USAMVB
from USAMV Bucharest compared to the red collection

Red flowers February 28 66.66% March 1
flowers variety from USAMV Bucharest. V11 peony from
USAMVB
Also, the growing period of the sprouts was collection
shorter at the red flowers variety compared to

the pink flowers variety from USAMV The duration of growing and blooming phenophases at white and
Bucharest. pink flowers varieties from SINGURENI forced in pots
The growth dynamics of the herbaceous peony 35

30
varieties in containerized and forced cultivation 25
is presented in Table 2. 20
15
An early vegetation start at V1, V6 and V8 and
days
10
a later occurrence at V3, V9 and V11 were 5
observed. 0
V1 V2 V4
The percentage of the vegetation start was the total duration of buds
starting
5 7 7
33.33% and 36.36% at V1 and V2 up to 95% at the duration of sprouts

growing
22 30 22
V7 and 100% at V6. the duration of blooming 5 4 5
The growth of sprouts started at V1 on 11.02, at the duration of a flower
on the plant
4 3 4
V6 on 10.02 and V8 on 10.02 and continued
until 20.02 at V10, and 01.03 at V11. Figure 8. The dynamics of growing and flowering at the
So, we are observing an early start of the cultivars with white and pink flowers from Singureni,
vegetation on the pink and red flowers varieties forced in pots in greenhouse, 2017
from USAMV Bucharest compared to the
white and pink flowers varieties from Regarding the growing of sprouts, the growth
Singureni. Also, the maximum vegetation lasted 22 days at V1 and V4, and 30 days at V2
starting percentage was observed at the pink (Figure 8).
flowers variety from USAMV Bucharest The flowering period as well as the duration of
compared to the white and pink flowers the flowers on the stem were short, between 3
varieties from Singureni and with red flowers and 5 days (Figure 8).
from USAMV Bucharest. We are concluding that the white flower variety
The beginning of buds vegetation lasted from Singureni had a shorter period for starting
between 5 days at V1 and 7 days at V2 and V4 the buds compared to the pink flowers variety
(Figure 8). from Singureni. Also, the sprouts development
536
period, the blooming period and the duration of peony plants at a higher cold season and
the flower on the plant is the same with the percentage of vegetation starting.
pink flower variety from Singureni.
The percentage of the vegetation starting was
33.33% at V1, and 36.36% at V2, up to 95% at
V7 and 100% at V6 (Figure 9).
Figure 10. Influence of the cold period on the vegetation

starting percentage at the variety with
white flowers by Singureni, 2017
The phenophases of flowering were only

conducted in 3 variants and 2 cultivars, V1 and
V2 cultivars with white flowers of Singureni,
and V4 cultivar with pink flowers of Singureni
Figure 9. The growing and blooming phenophases (Figure 11).
percentage of the herbaceous peony, forced in
greenhouse, 2017
The percentage of floral sprouts has been

shown in 7 variants out of 11, with values
between 22.22% at V6 up to 100% at V2.
The percentage of plants having floral sprouts
has been shown in 7 variants out of 11, with
values between 33.33% at V8 and 100% at V2
(Figure 9).
The percentage of flowering has been reached
in 3 variants out of 11, at V5 with 66.66%, and
100% at V1 and V2, respectively (Figure 9).
It is determined that the variety with pink
flowers from USAMV Bucharest had a Figure 11. Flowering phenophases of Singureni
maximum percentage related to the white and cultivars with white and pink flowers, 2017
pink from Singureni and red from USAMV
Bucharest. Also, a maximum percentage of The appearance of the bud occurred at V1 and
blooming sprouts and plants with blooming V4 on 18.02 and V2 on 24.02 (Figure 11).
sprouts was shown at the white flowers variety The opening of the flower was started at V1 on
from Singureni related to pink and red 14.03, at V2 on March 21, and on V4 on 17.03,
varieties. A maximum blooming percentage is and the blooming ended at V1 on 20.03, V2 on
observed at the white flowers variety from 25.03 and V4 on 22.03. (Figure 11).
Singureni related to pink and red varieties. So, we are observing that only the Singureni
The influence of the cold season on the flowers varieties were blooming and passed
percentage of vegetation starting at the through the blooming phenophases. An early
Singureni white flower variety is shown in blooming is observed at the white flowers
Figure 10. There is a positive influence of the variety compared to the pink flowers variety
cold period on the start of vegetation of the from Singureni.
cultivar with white flowers by Singureni. There Details regarding the growing phenophases are
is a correlation between the cold period and the found in the Figures 12-18.
start of vegetation: the higher the herbaceous
537
Figure 12. Floral sprouts, Figure 13. Floral sprouts, Figure 17. Flowering appearance of Singureni cultivar
white flowers cultivar, white flowers cultivar, with pink flowers, V5, 2017
Singureni, V1, 2017 Singureni, V2, 2017
Figure 14. Bud opening, Figure 15. The aspect of

pink flowers cultivar, flower and blooming,
Singureni, V5, 2017 pink flowers cultivar,
Singureni, V5, 2017
Figure 18. Flowering of white flowers cultivar,

Singureni, V1, 2017
The influence of the cold period on flowering

was observed in Figure 19, resulting in a
correlation between plant exposure to cold and
the beginning of inflorescence in herbaceous
peony, which shows how much the cold period
increases and the date of flowering increases
and vice versa decreases the period of exposure
to cold peony blooms earlier. So, the cold
Figure 16. Flower opening, white cultivar, V1, 2017 period has a positive influence on peony
bloom.
538
The earliness of the passing through the
growing phenophases was highlighted on all
varieties of herbaceous peony studied.
The varieties with pink and red flowers from
USAMV Bucharest had the earliest vegetation
starting, compared to the white and pink
flowers varieties from Singureni.
All varieties studied had a percentage of
vegetation starting over 50%.
The increase in the cold period positively
influences the percentage of starting the peony
vegetation.
Figure 19. Influence of the cold period on blooming The shortening of the cold period positively
in the varieties with white and pink flowers influences the early eruption of the herbaceous
by Singureni, 2017
peony, and the increase of the cold period
influences the early blooming.
After flowering, the plants continued to grow,
The early blooming and the earliness of the
forming rich bushes.
passing through the growing phenophases was
The largest begetting increases were recorded
highlighted at the white and pink flowers
at cultivars which red flowers from USAMV
variety from Singureni.
Bucharest and the cultivars which white
The white flowers variety from Singureni was
flowers from Singureni (Figures 20-21).
highlighted by a 100% percentage of blooming,
compared to the pink flowers variety from
Singureni (66.66%).
The earliness of the blooming was highlighted
at the white flowers variety from Singureni
(20.03), compared to the pink flowers variety
from Singureni (22.03).
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540
TREE SPECIES SELECTION GUIDELINES FROM

THE PERSPECTIVE OF BIOCLIMATIC LANDSCAPE DESIGN
Vladimir Ionuț BOC

Abstract
In landscape architecture, numerous researches carried out in different climatic regions around the world show that
arborescent vegetation has the most significant role in the improvement of microclimate. The study is focused on the
adaptation and integration of human bioclimatology and bioclimatic design principles in planting and landscape design
in order to develop tree species selection methods. Thus, in order to identify the most valuable tree species, a number of
criteria have been identified considering the impact of trees on the main factors influencing microclimate, such as heat,
wind speed, air humidity, light, air ionization and air quality. The results consist of a comparative analysis of tree
species from the temperate continental plain area. A ranking of the species was generated considering their potential
favourable impact on the microclimate as well as the eco-climatic resilience of the trees. The bioclimatic values of the
analysed species were attributed according to biological and ecological characteristics such as tree habitus, canopy
density, longevity, carbon storage capacity, air ionization level, resistance to wind, drought and air pollution, etc. The
tree species were assessed by researching specialized dendrological studies and through field observations. The
conclusions reveal the most valuable bioclimatic trees species, which are classified according to the way they can be
used in different types of green areas.
Key words: tree species selection, bioclimatic landscape design, urban microclimate improvement.
INTRODUCTION urban green areas, such as leaf area index

(LAI), carbon storage capacity, crown (canopy)
A comparative analysis of arborescent density, pollutants bioaccumulation capacity,
vegetation was initiated in order to identify air ionization, wind resistance, resistance to
which tree species presents the highest urban air pollution and tolerance to drought
potential for urban microclimate improvement conditions.
in the case of four different types of green
areas: A - shelterbelts, B - mobility areas, C - MATERIALS AND METHODS
green areas adjacent to buildings and D - green
cores (green areas over 1 ha). Classification of The assessed vegetation consist of frequently
green areas was carried out in previous studies encountered tree species in urban areas in the
(Boc, 2017) depending on the differential Bucharest-Ilfov metropolitan area were
impact of the vegetation on urban selected and a number of rarely encountered
microclimatic variables. species, which are well adapted to the local
In the present study, the analysed species are climatic conditions. The evaluated tree species
ranked according to the microclimatic impact were divided into three major categories:
and to the ecological and climatic resilience in - Conifers trees (persistent foliage);
the context of local conditions in Bucharest. - Deciduous trees I-II height level;
The hierarchy of the species has been - Deciduous trees III height level.
established taking into account specific criteria For each green area typology, the evaluation
that vary in each type of green area such as criteria were selected according to the
habitus, sunlight requirements, compatibility indicators mentioned in the introduction. For
with built environment or paved areas, each criterion, a score was attributed to the
emissions of volatile organic substances; but degree of bioclimatic value of the species.
also by general criteria, valid for all types of Thus, in the case of a low value, 1 point was
541
assigned, for an average value - 2 points, and - Low risk (3 points) = species without
for a high value - 3 points. Also, for each significant allergenic risk or moderately
criterion, the limits of each score have been set. allergenic dioecious species;
The values have been attributed by integrating 5. Carbon storage capacity - directly
research results of specialized studies, data proportional to the growth rate and LAI, except
provided by large tree nurseries in Europe and for VOC (Volatile organic compound)
visual observations in the north of Bucharest emissions in air polluted areas (species ranked
(such as Chitila, Bucureștii Noi, Domenii, according to Nowak, 2002; Nowak et al., 2007;
Băneasa, Herăstrău). The assessment methods Iliescu, 2005; Costăschescu et al., 2010; Van
and the source of the documentation are den Berk, 2017):
presented in the list of analysis criteria. Also, - Low capacity (1 point) = slow growth rate
for each criterion, the classification and ranking and low LAI;
of the assigned values are presented. - Average capacity (2 points) = average growth
The general criteria for analysis are divided rate and average LAI or fast growth rate and
into criteria for the bioclimatic impact of tree high LAI or slow growth rate and high LAI;
species (Boc, 2017) and criteria for climatic - High capacity (3 points) = fast growth rate
and ecological resilience in the case of the and high LAI;
temperate-continental urban microclimate, 6. Air ionization (species ranked according to
which is specific to Bucharest. Iliescu, 2005; Costăchescu et al., 2010;
The criteria for bioclimatic impact assessment Teodoreanu, 2011)
include: - Reduced ionization (1 point) = deciduous
1. The relative Leaf Area Index (LAI) - the species emitting positive ions;
species were ranked according to Asner et al. - Average ionization (2 points) = other
(2003) and to visual observations1: deciduous species;
- Low LAI (1 point) = LAI between 1 and 2; - High ionization (3 points) = coniferous trees
- Average LAI (2 points) = LAI from 3 to 4; and species with lanceolate leaves;
- High LAI (3 points) = LAI between 5 and 6; 7. Bioaccumulation of chemical pollutants
2. Canopy density (species ranked following (according to Nowak, 2002; Iliescu, 2005;
visual observations and according to Iliescu, Chira, Bolea, 2008; Costăchescu et al., 2010):
2005, Van den Berk, 2017): - Low capacity (1 point) = reduced
- Rare canopy (1 point) = Density below 40%; bioaccumulation compared to analysed species;
- Average canopy (2 points) = Canopy density - Medium capacity (2 points) = moderate
between 40 and 75%; bioaccumulation, close to the average of the
- Dense canopy (3 points) = Canopy density studied species;
over 75%; - High capacity (3 points) = high bioaccu-
3. Longevity of the species (species ranked mulation compared to other species;
according to Iliescu, 2005; Iliescu, 2006; Chira 8. Dust and particle retention capacity (species
and Bolea, 2008; Van den Berk, 2017): ranked according to Iliescu, 2005; Chira, Bolea,
- Low longevity (1 point) = under 50 years; 2008; Costăchescu et al., 2010; Van den Berk,
- Average longevity (2 points) = between 50 2017 and following field observations):
and 100 years; - Reduced capacity (1 point) = species with rare
- High longevity (3 points) = over 100 years; canopy and small leaves;
4. Security against allergens (species ranked - Medium capacity (2 points) = species with
following visual observations and according to rare canopy and large leaves or dense canopy
Iliescu, 2005; Carinanons and Casares-Porcel, and small leaves;
2011; Van den Berk, 2017): - High capacity (3 points) = species with large
- High risk (1 point) = strong allergic species; canopy and large leaves.
- Moderate risk (2 points) = strong allergenic The general criteria for the assessment of
dioecious species (only masculine specimens climatic and ecological resilience include (Ran-
are allergenic) or moderate allergenic species; king according to visual observations, Iliescu,
2005; Iliescu, 2006; Chira, Bolea, 2008;
Costăchescu et al., 2010; Van den Berk, 2017):
1
It was estimated for trees aged 20-40 years
542
9. Resistance to wind; Fraxinus excelsior, Fraxinus americana, Celtis
10. Resistance to drought; australis, Acer platanoides, Quercus rubra,
11. Resistance to air pollution. Carpinus betulus, Quercus robur, Ulmus
The following hierarchy applies to all criteria carpinifolia, Ulmus glabra;
concerning environmental and climatic - Deciduous tree species (height level III)
resilience: low resistance (1 point), medium recommended as dominants (Table 3):
resistance (2 points), high resistance (3 points). Elaeagnus angustifolia, Corylus avellana,
The specific criteria are listed below, separately Quercus pubescens, Morus alba, Pyrus
for each green zone typology. Following the communis, Pyrus nivalis, Fraxinus ornus,
evaluation of the species, the arithmetic mean Prunus cerasifera Pissardii, Salix babylonica,
of the values for each of the two categories of Salix matsudana, Acer tataricum, Cercis
criteria was established. Thus, the final score canadensis, Sorbus aria, Ginkgo biloba,
resulted by achieving the arithmetic mean of Quercus cerris.
the microclimatic impact general score and the
climatic and ecological resilience score. Zone A - Protection green areas. Shelterbelts
Species were ranked according to the final In the case of relatively narrow urban
score (global average) according to which they shelterbelts, the use of dense canopy species
were grouped into three major categories and a predominantly vertical habitus is
within each green zone typology: recommended, allowing the formation of
Species recommended as dominants - high compact planting compositions. Considering
bioclimatic impacts and high climatic and the high density of plantations, the adaptation
ecological resilience (impact and resilience of the species for partly shaded areas is also an
scores ≥ 2.25, species which obtained over all important condition for climate and ecological
the criteria above 1 point) - total score resilience. Following the analysis, in the group
represented in green in Tables 1, 2, 3, 4 and 5; of coniferous trees, three species obtained
Complementary species - average bioclimatic scores from 2 up to all criteria and, implicitly,
impact values and climate and ecological the highest score. Thus, after the evaluation, the
resilience (impact and resilience scores ≥ 1.75, species were divided into the following three
species which have received no more than two categories:
marks of 1 point for the general criteria or a Coniferous trees (Table 5):
maximum of 1 point for the specific criteria) - -Recommended species: Pseudotsuga menziesii
represented in yellow in the tables; ssp. glauca, Juniperus virginiana, Pseudotsuga
Not recommended species (average impact and menziesii ssp. menziesii;
resilience scores ≤ 1.75, species which obtained - Not recommended species: Pinus silvestris,
more than two marks of 1 point on the analysed Pinus strobus.
criteria) - represented in orange. Deciduous tree species (height level I-II)
(Table4):
RESULTS AND DISCUSSIONS - Recommended species: Carpinus orientalis,
Fraxinus angustifolia, Ulmus glabra, Celtis
Bioclimatic value of tree species according to australis, Fraxinus excelsior, Fraxinus
general criteria americana, Carpinus betulus, Ulmus
The species identified with the highest carpinifolia, Maclura pomifera, Acer
bioclimatic potential following assessment platanoides, Ginkgo biloba2, Quercus rubra;
based only on the general criteria are: - Not recommended species: Populus tremula,
- Coniferous tree species recommended as Populus x canadensis, Platanus x hybrida,
dominants (Table 1): Pseudotsuga menziesii Populus alba, Betula pendula, Robinia
ssp. glauca, Juniperus virginiana, Pseudotsuga pseudacacia, Paulownia tomentosa, Ailanthus
menziesii ssp. menziesii, Picea pungens, Abies altissima.
concolor; Deciduous tree species (height level III)
- Deciduous tree species (height level I-II) (Table 5):
recommended as dominants (Table 2):
Carpinus orientalis, Fraxinus angustifolia, 2
Ginkgo biloba was placed in the deciduous trees
category considering it is not an evergreen coniferous.
543
- Recommended species: Corylus avellana, - Not recommended species: Salix babylonica,
Elaeagnus angustifolia, Crataegus monogyna, Ulmus pumila, Prunus ʻAccoladeʼ, Acer
Fraxinus ornus, Prunus cerasifera ʻPissardiiʼ, negundo, Malus baccata ʻStreet Paradeʼ,
Sorbus aria; Albizzia julibrissin, Prunus serrulata ʻKanzanʼ,
- Not recommended species: Salix babylonica, Malus silvestris.
Acer negundo, Prunus avium, Prunus cerasus,
Malus baccata, Malus domestica, Prunus Zone C - Buffer green areas.
ʻAccoladeʼ, Koelreuteria paniculata, Malus Plantations adjacent to the buildings
silvestris, Prunus domestica, Albizzia In the green areas planted adjacent to the
julibrissin, Prunus serrulata ʻKanzanʼ; buildings, the density of the vegetation should
be high and the planted areas consist of
Zone B - Mobility green areas. Greenways, relatively narrow strips (Boc, 2017). In this
trees alignments and planted platforms case, we will choose species with vertical
In the case of the green areas designed for habitus compatible with the built environment
mobility, the density of planted areas is from the point of view of tree crowns, root
relatively low and the main bioclimatic role of systems and fruits that may affect paved
vegetation is to provide shade and retain the surfaces. Subzone C1 is a type of buffer green
heat reflected by the paved surfaces. In this infrastructures located near the southern
regard, species with a dense and wide canopy facades. Subzone C2, situated near the northern
will be chosen to cover larger areas. Regarding facades and subzone C3, located between
the ecological resilience of species, it is buildings, it was taken into account the
essential to choose species compatible with complementary criterion of resistance to semi-
road and pedestrian infrastructure from the shade conditions. The following species have
point of view of the root system and fruit that been hierarchized for subzone C1:
can affect the paved areas. Deciduous tree species (height level I-II)
In zone B, the coniferous tree species were not (Table 4):
taken into account as the ability to provide - Recommended species: Fraxinus angustifolia,
shade and resistance to air pollution are Carpinus orientalis, Carpinus betulus,
generally low. Considering the results, the Fraxinus excelsior, Fraxinus americana, Acer
following species for mobility areas were platanoides, Celtis australis, Quercus rubra,
identified: Ulmus carpinifolia, Ginkgo biloba, Quercus
Deciduous tree species (height level I-II) cerris, Quercus robur;
(Table 4): - Not recommended species: Ulmus glabra,
- Recommended species: Carpinus orientalis, Acer pseudoplatanus, Salix alba, Populus
Fraxinus angustifolia, Fraxinus excelsior, Acer tremula, Populus nigra, Platanus x hybrida,
platanoides, Carpinus betulus, Fraxinus Aesculus hippocastanum, Acer campestre,
americana, Celtis australis, Quercus rubra, Populus x canadensis, Paulownia tomentosa,
Ginkgo biloba, Quercus robur, Quercus cerris, Populus alba, Betula pendula, Acer negundo,
Ulmus carpinifolia; Robinia pseudacacia, Ailanthus altissima;
- Not recommended species: Ulmus glabra, Deciduous tree species (height level III)
Salix alba, Populus tremula, Paulownia (Table 5):
tomentosa, Populus nigra, Aesculus - Recommended species: Elaeagnus angustifolia,
hippocastanum, Sophora japonica, Populus Corylus avellana, Pyrus communis, Pyrus nivalis,
alba, Populus x canadensis, Betula pendula, Fraxinus ornus, Prunus cerasifera ʻPissardiiʼ,
Ailanthus altissima, Robinia pseudacacia; Sorbus aria;
Deciduous tree species (height level III) - Not recommended species: Salix babylonica, Acer
(Table 5): tataricum, Acer negundo, Malus baccata, Malus
domestica, Prunus ʻAccoladeʼ, Prunus domestica,
- Recommended species: Quercus pubescens,
Ulmus pumila, Malus silvestris, Albizzia julibrissin,
Elaeagnus angustifolia, Corylus colurna, Prunus serrulata ʻKanzanʼ.
Prunus cerasifera ʻPissardiiʼ, Fraxinus ornus, The following semi-shade species, identified
Sorbus aria, Acer tataricum; for subzones C2 and C3, have been identified:
Coniferous trees (Table 5)
544
Since coniferous trees are compatible with the potential belong to the deciduous tree group
built environment and the rest of the analysis (height level I-II) (Table 1). The genera
criteria are similar to those in the case of the characterized by the highest values are
shelterbelts, then the results from zone A are Fraxinus, Carpinus, Celtis and Acer, which are
valid also for subzones C2 and C3. compatible with all four types of green areas
Deciduous tree species (height level I-II) (Table 4).
(Table 4): Among the deciduous tree of height level III,
- Recommended species: Carpinus orientalis, there is a high degree of compatibility with all
Fraxinus angustifolia, Carpinus betulus, types of green areas in the case of the species
Fraxinus excelsior, Fraxinus americana, Celtis Eleagnus angustifolia, Fraxinus ornus, Prunus
australis, Acer platanoides, Ginkgo biloba, cerasifera and Sorbus aria (Table 5).
Ulmus carpinifolia, Quercus rubra; From the coniferous category, the species with
- Not recommended species: Tilia tomentosa, the highest score are Juniperus virginiana and
Acer pseudoplatanus, Ulmus glabra, Salix Pseudotsuga sp. (Table 5). It should be noted
alba, Quercus cerris, Quercus robur, Gleditsia that although they have a high degree of
triacanthos, Aesculus hippocastanum, Sophora adaptability to environmental conditions, the
japonica, Populus tremula, Populus nigra, two coniferous species are rarely used in
Platanus x hybrida, Populus x canadensis, Bucharest.
Paulownia tomentosa, Liriodendron tulipifera,
Populus alba, Acer negundo, Robinia REFERENCES
pseudacacia, Ailanthus altissima;
Deciduous tree species (height level III) Asner G.P., Scurlock J.M.O., Hicke J., 2003. Global
(Table 5): synthesis of leaf area index observations:
implications for ecological and remote sensing
- Recommended species: Elaeagnus studies Global Ecology & Biogeography, Blackwell
angustifolia, Corylus avellana, Fraxinus ornus, Publishing Ltd 12, 191-205.
Populus simonii, Prunus cerasifera Pisardii, Boc V.I., 2017. Introducing the bioclimatic security
Sorbus aria, Prunus padus; concept in green infrastructure planning and design,
- Not recommended species: Pyrus communis, ISHS Acta Horticulturae, vol. VI International
Conference on Landscape and Urban Horticulture
Pyrus nivalis, Salix matsudana, Salix 1189, 235-240.
babylonica, Prunus avium, Ulmus pumila, Bolea V., Chira D., 2008. Flora indicatoare a poluării,
Prunus Accolade, Acer negundo, Koelreuteria Editura Silvică, București.
paniculata, Prunus domestica, Malus silvestris, Carinanos P., Casares-Porcel M., 2011. Urban green
Prunus serrulata Kanzan, Albizzia julibrissin. zones and related pollen allergy: A review. Some
guidelines for designing spaces with low allergy
impact, Landscape and Urban Planning 101, 205-214.
Zone D - Green cores Costăchescu C., Dănescu F., Mihăilă E., 2010. Perdele
Green cores include consistent planted areas forestiere de protecţie, Editura Silvică, Bucureşti.
and do not impose specific restrictions Iliescu A.F., 2005. Cultura arborilor şi arbuştilor
regarding road infrastructure and buildings. ornamentali, Editura Ceres, Bucureşti.
Iliescu A.F., 2006. Arhitectură peisageră, Editura Ceres,
Thus, in this case it is recommended to choose Bucureşti.
the dominant species from among the trees with Nowak D.J., 2002. The Effects Of Urban Trees On Air
the most bioclimatic impact resulting from the Quality. USDA Forest Service, Syracuse, New York.
assessment based on general criteria. In this Nowak D.J., Hoehn R., Crane D.E., 2007. Oxygen
case, other species except those mentioned in Production by Urban Trees in the United States,
Arboriculture & Urban Forestry 2007. 33 (3):220-
the selection analysis may be inserted 226.
separately, provided that the recommended Teodoreanu E., 2002. Bioclimatologie umană, Editura
dominant species are used in the planting Academiei Române, București.
composition. Van den Berk Nurseries, 2017.
https://www.vdberk.co.uk/trees
CONCLUSIONS
Following the ranking of the species, it was

found that trees with the highest bioclimatic
545
Table 1. Assessment and ranking of coniferous tree species according to the relative bioclimatic value
FACTORS INFLUENCING THE BIOCLIMATIC IMPACT OF DENDROLOGIC VEGETATION
GENERAL CRITERIA IMPACT ON EOLIEN FACTORS IMPACT ON AEROCHEMICAL AND AEROELECTRIC FACTORS CLIMATE AND ECOLOGICAL RESILIENCE
Accumulat-
Dust Resistance
Estimated Canopy Allergens Carbon Air ion of Wind Drought Total 1 - Total 2 -
Coniferous tree species Longevity particles to air TOTAL
LAI density security storage ionization chemical resistance resistance Impact Resilience
retention pollution
pollutants
Pseudotsuga menziesii ssp. glauca 3 2 3 3 2 3 2 3 3 3 3 2,63 3,00 2,81
Juniperus virginiana 2 3 3 3 2 3 2 2 3 3 2 2,50 2,67 2,58
Pseudotsuga menziesii ssp. menziesii 3 2 3 3 2 3 2 3 3 2 2 2,63 2,33 2,48
Picea pungens 2 3 2 2 2 3 3 2 3 2 2 2,38 2,33 2,35
546
Abies concolor 2 3 3 2 2 3 2 2 3 2 1 2,38 2,00 2,19
Abies nordmanniana 3 2 3 2 2 3 3 3 3 1 1 2,63 1,67 2,15
Abies alba 3 2 3 3 2 3 2 3 3 1 1 2,63 1,67 2,15
Cupressocyparis leylandii 2 3 2 2 2 3 3 3 1 2 2 2,50 1,67 2,08
Thuja plicata 3 3 2 2 3 3 2 3 1 1 2 2,63 1,33 1,98
Picea abies 3 2 3 3 2 3 3 2 2 1 1 2,63 1,33 1,98
Chamaecyparis lawsonianna 2 3 2 2 2 3 3 3 1 1 2 2,50 1,33 1,92
Pinus nigra 2 2 3 2 2 3 1 2 2 2 1 2,13 1,67 1,90
Pinus strobus 2 2 3 2 2 3 2 2 2 1 1 2,25 1,33 1,79
Pinus silvestris 2 1 3 2 2 3 1 1 2 2 1 1,88 1,67 1,77
Relative bioclimatic value
1 - low 2 - medium 3 - high no data

Table 2. Assessment and ranking of deciduous tree species (height level I-II) according to the relative bioclimatic value
Accumulat-
Dust Resistance
Deciduous tree species Estimated Canopy Allergens Carbon Air ion of Wind Drought Total 1 - Total 2 -
Longevity particles to air TOTAL
(Height level I-II) LAI density security storage ionization chemical resistance resistance Impact Resilience
retention pollution
pollutants
Tilia tomentosa 3 3 3 1 2 2 3 3 3 3 3 2,50 3,00 2,75
Carpinus orientalis 3 3 3 2 2 2 3 3 3 3 2 2,63 2,67 2,65
Fraxinus angustifolia 3 2 3 2 3 2 3 3 3 2 3 2,63 2,67 2,65
Fraxinus excelsior 3 2 3 2 3 2 3 3 3 2 3 2,63 2,67 2,65
Fraxinus americana 2 2 3 2 3 2 3 3 3 2 3 2,50 2,67 2,58
Celtis australis 2 2 3 2 2 2 2 2 3 3 3 2,13 3,00 2,56
Acer platanoides 3 3 2 2 3 2 3 3 2 3 2 2,63 2,33 2,48
Carpinus betulus 3 3 3 2 2 2 3 3 3 2 2 2,63 2,33 2,48
Quercus rubra 2 2 3 2 2 3 2 2 3 2 3 2,25 2,67 2,46
Acer pseudoplatanus 3 3 2 2 3 2 3 2 3 1 3 2,50 2,33 2,42
Salix alba 2 2 3 2 3 3 3 2 3 1 3 2,50 2,33 2,42
Ulmus carpinifolia 2 2 3 2 2 2 2 3 2 3 2,14 2,67 2,40
Ulmus glabra 2 2 3 2 2 2 2 3 2 3 2,14 2,67 2,40
Maclura pomifera 2 2 2 2 3 2 2 2 3 3 2,14 2,67 2,40
Ginkgo biloba* 2 2 3 3 2 3 2 2 2 3 2,43 2,33 2,38
547
Quercus robur 2 2 3 2 2 3 3 2 3 2 2 2,38 2,33 2,35
Populus nigra 3 2 2 1 3 3 3 2 3 2 2 2,38 2,33 2,35
Populus tremula 2 2 2 1 3 3 3 3 3 1 3 2,38 2,33 2,35
Quercus cerris 2 2 3 2 2 3 2 3 2 2 2,29 2,33 2,31
Platanus hybrida 3 2 3 1 3 2 2 2 2 3 2,29 2,33 2,31
Sophora japonica 2 2 1 3 3 3 2 2 2 3 2,29 2,33 2,31
Populus canadensis 2 1 2 1 3 3 3 2 3 1 3 2,13 2,33 2,23
Populus alba 2 1 2 1 3 3 3 2 2 2 3 2,13 2,33 2,23
Aesculus hippocastanum 3 3 3 1 2 2 3 2 2 2 2,43 2,00 2,21
Gleditsia triacanthos 1 1 3 3 2 1 2 1 2 3 3 1,75 2,67 2,21
Quercus palustris 2 2 3 2 2 1 2 3 1 3 2,00 2,33 2,17
Juglans regia 2 2 3 2 2 3 2 2 3 1 2 2,20 2,00 2,10
Robinia pseudacacia 1 1 2 1 1 3 1 2 3 3 1,43 2,67 2,05
Paulownia tomentosa 2 1 2 3 3 3 3 1 1 3 2,43 1,67 2,05
Liriodendron tuilipifera 2 2 3 3 3 2 2 2 2 1 2 2,38 1,67 2,02
Acer campestre 2 2 2 2 2 2 2 2 2 2 2 2,00 2,00 2,00
Tilia platyphyllos 2 2 3 1 2 1 3 2 2 2 2 2,00 2,00 2,00
Betula pendula 1 1 2 1 1 3 3 1 2 2 3 1,63 2,33 1,98
Ailanthus altissima 2 1 1 2 2 1 2 2 2 2 3 1,63 2,33 1,98

Table 3. Assessment and ranking of deciduous tree species (height level III) according to the relative bioclimatic value
GENERAL CRITERIA IMPACT ON EOLIEN FACTORS IMPACT ON AEROCHEMICAL AND AEROELECTRIC FACTORS CLIMATE AND ECOLOGICAL RESILIENCE
Accumulat-
Dust Resistance
Deciduous tree species Estimated Canopy Allergens Carbon Air ion of Wind Drought Total 1 - Total 2 -
Longevity particles to air TOTAL
(Height level III) LAI density security storage ionization chemical resistance resistance Impact Resilience
retention pollution
pollutants
Elaeagnus angustifolia 2 2 3 2 2 3 3 2 3 3 3 2,38 3,00 2,69
Corylus avellana 2 3 3 2 2 3 3 3 2 3 2,57 2,67 2,62
Quercus pubescens 2 2 3 2 2 2 3 3 3 2,17 3,00 2,58
Crataegus monogyna 2 2 3 2 1 2 2 3 3 3 2,00 3,00 2,50
Populus simonii 3 3 2 2 3 3 3 2 3 1 3 2,63 2,33 2,48
Morus alba 2 2 3 2 2 3 2 2 3 3 2,29 2,67 2,48
Pyrus communis 2 2 3 3 2 2 2 3 3 2 2,29 2,67 2,48
Pyrus nivalis 2 2 3 3 2 2 2 3 3 2 2,29 2,67 2,48
Fraxinus ornus 2 2 3 2 3 2 3 3 2 3 2 2,50 2,33 2,42
Prunus cerasifera Pisardii 2 3 2 3 2 2 3 2 2 3 2,43 2,33 2,38
Salix babylonica 2 2 2 2 3 3 3 2 3 2 2 2,38 2,33 2,35
Salix matsudana 2 2 1 2 3 3 3 2 3 2 2 2,25 2,33 2,29
Acer tataricum 2 2 2 3 2 2 2 2 2 2 3 2,13 2,33 2,23
548
Cercis canadensis 2 2 1 3 2 2 3 2 3 2 2,14 2,33 2,24
Sorbus aria 2 2 3 3 2 2 2 2 2 2 2,29 2,00 2,14
Prunus padus 2 3 2 2 2 3 1 2 2 2,33 1,67 2,00
Ulmus pumila 2 1 2 2 2 1 2 2 3 1,67 2,33 2,00
Acer negundo 2 2 1 2 2 2 3 2 1 2 3 2,00 2,00 2,00
Prunus Accolade 1 1 2 2 2 2 1 2 2 3 1,57 2,33 1,95
Catalpa bignonioides 2 2 2 3 2 2 2 2 1 2 2,14 1,67 1,90
Prunus avium 1 2 2 2 2 1 2 2 2 2 1,71 2,00 1,86
Albizzia julibrissin 1 1 1 3 1 2 2 2 2 2 1,57 2,00 1,79
Koelreuteria paniculata 2 2 1 2 2 2 2 2 1 2 2 1,88 1,67 1,77
Prunus cerasus 1 2 2 2 2 2 2 2 1 2 1,86 1,67 1,76
Malus baccata 1 2 2 2 1 2 2 2 1 2 1,71 1,67 1,69
Malus domestica 1 2 2 2 1 2 2 2 1 2 1,71 1,67 1,69
Prunus domestica 1 2 2 3 2 2 2 1 2 1 2,00 1,33 1,67
Prunus serrulata Kanzan 1 2 2 1 2 2 1 2 1 1,67 1,50 1,58
Malus silvestris 1 2 2 2 1 1 1 2 1 2 1,43 1,67 1,55

Table 4. Compatibility level between deciduous tree species (height level I-II) and green areas typologies
Subzone Subzone
Deciduous trees height I-II Zone A Zone B Zone D
C1 C2, C3
Acer campestre
Acer negundo
Acer platanoides
Acer pseudoplatanus
Aesculus hippocastanum
Ailanthus altissima
Betula pendula
Carpinus betulus
Carpinus orientalis
Celtis australis
Fraxinus americana
Fraxinus angustifolia
Fraxinus excelsior
Ginkgo biloba*
Gleditsia triacanthos
Juglans regia
Liriodendron tuilipifera
Maclura pomifera
Paulownia tomentosa
Platanus hybrida
Populus alba
Populus canadensis
Populus nigra
Populus tremula
Quercus cerris
Quercus palustris
Quercus robur
Quercus rubra
Robinia pseudacacia
Salix alba
Sophora japonica
Tilia platyphyllos
Tilia tomentosa
Ulmus carpinifolia
Ulmus glabra
Compatibility level Low Medium High
549
Table 5. Compatibility level between coniferous tree species and green areas typologies (up and down)
Subzone Subzone
Coniferous trees Zone A Zone B Zone D
C1 C2, C3
Abies alba
Abies concolor
Abies nordmanniana
Chamaecyparis lawsonianna
Cupressocyparis leylandii
Juniperus virginiana
Picea abies
Picea pungens
Pinus nigra
Pinus silvestris
Pinus strobus
Pseudotsuga menziesii ssp. glauca
Pseudotsuga menziesii ssp. menziesii
Thuja plicata
Subzone Subzone
Deciduous trees height III Zone A Zone B Zone D
C1 C2, C3
Acer negundo
Acer tataricum
Albizzia julibrissin
Catalpa bignonioides
Cercis canadensis
Corylus avellana
Corylus colurna
Crataegus monogyna
Elaeagnus angustifolia
Fraxinus ornus
Koelreuteria paniculata
Malus baccata
Malus domestica
Malus silvestris
Morus alba
Populus simonii
Prunus Accolade
Prunus avium
Prunus cerasifera Pisardii
Prunus cerasus
Prunus domestica
Prunus padus
Prunus serrulata Kanzan
Pyrus communis
Pyrus nivalis
Quercus pubescens
Salix babylonica
Salix matsudana
Sorbus aria
Ulmus pumila
Compatibility level Low Medium High
550
ANALYSIS OF CHANGES IN GROWING DEGREE-DAY VALUES

BY ALTITUDE: OIL ROSE (ROSA DAMASCENA MILL.) CASE
Ali YUCEL1, Atılgan ATILGAN2, Cagatay TANRIVERDI3, Hasan ÖZ2

1
Osmaniye Korkut Ata University, Osmaniye Vocational School, 80000 Osmaniye, Turkey
2
Süleyman Demirel University, Faculty of Agriculture, Agricultural Structure and Irrigation
Department, 32260 Isparta,Turkey
3
Kahramanmaras Sutcu Imam University, Faculty of Agriculture, Biosystem Department, 46000
Kahramanmaras,Turkey
Abstract
Times such as optimum sowing, planting, germination time and duration, fertilization, agricultural pest control,
irrigation time, harvest, plant growing stages can be determined by using the Growing Degree-Day (GDD) values. The
Isparta region has an ideal climatic area for oil rose growing. For this purpose, meteorological stations were chosen as
the study area for Isparta provinces and districts having the most suitable growing ecology in oil rose cultivation. Oil
rose is a perennial plant with an active growing period of about 120 days depending on the phenological periods.
Different GDD values for different phenological periods were acquired when the GDD values for the active growing
period of the oil rose plant were correlated with the elevations of meteorological stations at different locations. It was
determined that the study area was divided into 4 groups by the statistical Duncan test, which was made according to
the growing degree-day values and optimum temperatures. It was determined that the Sütçüler district is the most suited
growing area for the oil rose plant followed by Atabey, Eğirdir, Isparta, Keçiborlu, Senirkent and Uluborlu.
Key words: Isparta, phenological periods, rose plant, temperature.
INTRODUCTION intensity of oil rose. Oil rose plants are quite

resistant to cold weather during the winter
Volatile oils are obtained from the flowers, when it sheds its leaves. Resting of mature
leaves, fruits, seeds and roots of plants. Rose is plants during the winter season helps bud
the most popular and most important among all development and differentiation. However, it is
volatile oil plants grown in Turkey (Rosa very sensitive to low temperatures and frost
damascena Mill.). Turkey is the biggest global during the budding period (starting in March).
producer of oil rose (Ikiz and Demircan, 2013). It is desired that the temperature varies between
Roses have an important place among 5-20oC during the flowering period of oil rose.
medicinal and aromatic plants. Some genotypes Volatile oil content decreases when nighttime
of Rosa damascena have been grown for temperatures drop below 5oC or when daytime
industrial purposes in Turkey (Region of temperatures exceed 20oC (Baydar and Baydar,
Lakes) since 1888. Oil production started in 2005; Sangwan et al., 2001).
1892 and it has been processed as an industrial Climate data should be analyzed accurately
plant since 1935 (Özçelik et al., 2013). Turkey since plant growth depends solely on climate
and Bulgaria are the countries with the highest conditions. Changes in temperature and rainfall
amount of oil rose growth in the world. These play a determining role on yield especially
two countries meet 80% of the total oil rose during phenological periods of plant growth.
production in the world. According to 2016 Plant growing stages such as optimal
data, a total production of 12,267 tons has been plantation, germination time, fertilization, pest
completed on 2,975.3 hectares in Turkey (Tuik, control, irrigation time and harvesting period,
2017). may be determined using the degree day
Temperature and humidity are among the most method. The purpose of the study was to
important factors with impacts on the flowering determine and interpret the relations between
551
the altitude values of locations included in the study have been given in Table 1. Average
study area for the oil rose plant and the altitude of the province was about 1,065 m
calculated GDD values. (Anonymous, 2018).
Temperature values given in Table 2 (Baydar
MATERIALS AND METHODS and Kazaz, 2013) have been used for
calculating the GDD values of oil rose in
In the study, long term maximum and different phenological periods. Changes
minimum daily temperature values with between the calculated GDD values and the
different record lengths, measured at different altitudes of meteorological stations (H, m) were
meteorological stations (16 in total) in the tested by way of regression analysis. Statistical
Isparta city limits have been used (Table 1). Duncan test was used for grouping the GDD
Since meteorological stations in the city of values of oil rose plant calculated in different
Isparta are built in districts or close-by regions, phenological periods and the altitudes of
they are known by the names of these districts. meteorological stations.
Properties for the 16 meteorological stations in
the city of Isparta and its districts used in the
Table1. Characteristics of Meteorological Stations used in the study

Meteorological Years Period duration Latitude Longitude Altitude (m)
stations (years)
Aksu 1983 – 2003 21 37°47' 31°04' 1,240
Atabey 1968 – 2015 48 37°57' 30°38' 1,000
Bağkonak 1987 – 1996 10 38°14' 31°17' 1,397
Barla 1987 – 1992 6 38°01' 30°78' 1,085
Eğirdir 1968 – 2015 48 37°50' 30°52' 917
Gelendost 1983 – 1988 6 38°07' 31°01' 952
Isparta 1929 – 2015 87 37°47' 30°34' 997
Kasımlar 1987 – 1993 7 37°53' 31°19' 1,070
Keçiborlu 1971 – 1990 20 37°57' 30°18' 996
Kumdankı 1984 – 1995 12 38°32' 30°97' 1,029
Senirkent 1970 – 2015 46 38°06' 30°33' 959
Sütcüler 1968 – 2015 48 37°30' 30°59' 975
Sarkıkaraağaç 1976 – 2015 40 38°05' 31°22' 1,180
Uluborlu 1968 – 2015 48 38°05' 30°27' 1,025
Yalvaç 1972 – 2015 44 38°16' 31°10' 1,096
Yenisar-Bademli 1983 – 1994 12 37°42' 31°23' 1,183
Table 2. Optimum Temperature Requirements for Different Phenological Periods of Oil Rose
Temperature
Phenological Periods Date Period duration (days)
(oC)
Bud break 5 – 10 1 March– 15 March 15
Shoot bud 10 – 15 16 March – 31 March 16
Leaf and Flowering bud 15 – 18 1 April – 30 April 30
Flowering and harvesting 15 – 25 1 May – 30 June 61
Total 122
Growing Degree-Day (GDD) Method values have been used in agricultural studies.
Agriculturally cultivated plants benefit Growing Degree Day (GDD) method is the
differently from the temperature in each growth most frequently used method. In this method,
period. Temperature is one of the most calculations are made based on the daily
important meteorological factors with impacts maximum (TMax) and daily minimum (TMin)
on plant growing. Various methods for values measured in meteorological stations.
estimating plant growing using temperature GDD values are calculated using the below
552
equation (McMaster and Wilhelm, 1997; Variance analyses. It takes into account the
Kadıoğlu and Şaylan, 2001; Snyder et al., positions of the averages according to their
2001; Matzarakis et al., 2007; Rulm et al., magnitudes when comparing group averages
2010; de Souza et al., 2011). for this purpose. The minimum difference
between two group averages in this method is
TMax TMin calculated by way of Duncan table. Group
n
averages are evaluated according to the
BDG TO (1)
İ 1 2 distances between them when ordered based on
magnitude and it is one of the most frequently
where: TMax stands for the daily maximum used methods (Duncan, 1955; Harter, 1960; Efe
temperature value (°C), TMin stands for the et al., 2000).
daily minimum temperature value (°C),
whereas TO denotes the temperature value RESULTS AND DISCUSSIONS
suggested for different phenological periods for
the rose plant (°C) and n represents the yearly Altitude and temperature values acquired from
number of days. In case TMax TMin 2 TO measurement stations at 16 different locations
growing degree-day (GDD) value is calculated. in the city of Isparta and its districts have been
It means that growing of the plant is used in the study. Figure 1 was prepared
determined. On the contrary, when depicting the graphical relations between the
TMax TMin 2 TO growing degree-day GDD values for different phenological periods
(GDD) not calculated. It means that there is no of the oil rose plant and the elevations of the
growing in the plant (McMaster and Wilhelm, districts. Oil rose is a perennial plant with an
1997; Kadıoğlu and Şaylan, 2001; Snyder et active growing period of about 120 days
al., 2001; Matzarakis, et al., 2007; Rulm et al., depending on the phenological periods (Baydar
2010; de Souza et al., 2011). and Kazaz, 2013). Different GDD values for
different phenological periods were acquired
Statistical methods when the GDD values for the active growing
Regression analysis. The objective with the period of the oil rose plant were correlated with
dependent variable to be determined is to put the elevations of meteorological stations at
forth an estimation method for determining the different locations. The average elevation of the
relationship between one or more independent city of Isparta is 1,065 m and Sütçüler region
variables. The method developed for this with an elevation of 975 m was determined as
purpose can be used to make estimations. The the district with the highest GDD value during
general equation is expressed with the the bud break period. Whereas the lowest GDD
following equality. values during the bud break period were
determined in Yenişar Bademli. It was
Y a b.x (2) determined upon an examination of the shoot
bud period that the Sütçüler region had the
where: Y is the dependent variable, x is the highest GDD values during this period as well.
independent variable, whereas a and b are The lowest GDD values were determined in the
equation regression coefficients. Regression Gelendost district. No linear relationship was
analysis statistical fit is evaluated by; determined between the phenological
correlation coefficient (r), F test and probability temperature demands of the oil rose plant and
(p) (Haan, 1977; Helsen ve Hirsch, 1993; the elevation values of the districts during the
Shammugasundram, 2012). shoot bud period as well. It was determined that
the GDD values during the leaf and flowering
Duncan Multiple Comparison Test: It is one bud periods are above 70 for the Sütçüler
of the most frequently used methods in region. The lowest GDD value was observed at
agricultural studies. It is used for determining Yenişar Bademli. Sütçüler region was observed
whether there are statistically significant to have the highest GDD value during the final
differences between the variables obtained as a phenological period of oil rose which is
result of statistical analyses such as Regression, flowering and harvesting period. It can again be
553
observed in this stage that GDD values do not district is the best in the study area with regard
have a linear relationship with elevation. to the temperature demands of the oil rose plant
Yenişar Bademli had the lowest GDD values during its phenological periods. Local
during the flowering and harvest period. producers in the region along with Baydar and
As put forth by Serter (2004), Nield and Smith Kazaz (2013) indicate that Keçiborlu, Isparta
(1997), determined during their study on the central villages, Atabey, Eğirdir, Uluborlu,
maturing of corn plant at different locations, Senirkent and their environs, are the best areas
maturity times varied according to locations. for oil rose. Therefore, it was determined that
Thus, the GDD values acquired for 16 different all areas, excluding Sütçüler, were determined
locations in the study area were different for oil to be compatible with regard to GDD values
rose plant. It can be indicated that Sütçüler and elevation.
5°C 6°C 7°C 8°C

9°C 10°C Altitude
80 1600
Growing Degree-Day Values (°C*day)
70 1400
60 1200
50 1000
Altitude (m)
40 800
30 600
20 400
10 200
0 0
Barla
Aksu
Sütçüler
Kumdanlı
Kasımlar
Isparta
Uluborlu
Bağkonak
Eğirdir
Senirkent
Keçiborlu
Yalvaç
Şarkıkaraağaç
Atabey
Yenisar-Bademli
Gelendost
a. Bud break
10°C 11°C 12°C 13°C

14°C 15°C Altitude
40 1600
35 1400
30 1200
Altitude (m)
25 1000
20 800
15 600
10 400
5 200
0 0
Barla
Uluborlu
Kumdanlı
Kasımlar
Aksu
Bağkonak
Eğirdir
Senirkent
Sütçüler
Keçiborlu
Isparta
Yalvaç
Atabey
Şarkıkaraağaç
Yenisar-Bademli
Gelendost
b. Shoot bud
554
15°C 16°C 17°C 18°C Altitude
80 1600

70 1400
60 1200
50 1000
Altitude (m)
40 800
30 600
20 400
10 200
0 0
Kasımlar
Barla
Aksu
Sütçüler
Isparta
Uluborlu
Kumdanlı
Yalvaç
Bağkonak
Eğirdir
Senirkent
Keçiborlu
Atabey
Şarkıkaraağaç
Yenisar-Bademli
Gelendost
c. Leaf and flowering bud
15°C 16°C 17°C 18°C

19°C 20°C 21°C 22°C
23°C 24°C 25°C Altitude
800 1600
Growing Degree-Day Values (BDG, °C*day)
700 1400
600 1200
Altitude (m)
500 1000
400 800
300 600
200 400
100 200
0 0
Barla
Aksu
Bağkonak
Senirkent
Sütçüler
Keçiborlu
Isparta
Uluborlu
Kumdanlı
Kasımlar
Yalvaç
Eğirdir
Atabey
Şarkıkaraağaç
Yenisar-Bademli
Gelendost
d. Flowering and harvesting
Figure 1. Relationship between GDD values and altitude in different phenological periods of oil rose plant
The relationships between the elevation (H, m) examined by way of regression analysis and the
values for the different meteorological stations statistical values for which two-parameter
at different locations in the study area and the parabolic equation (GDD a b.H c.H 2 ) has been
GDD values calculated for the different developed are given in Table 3.
phenological periods of oil rose plant, were
555
Table 3. Relations between GDD values, altitude and statistical results
Suggested Temperature Equation Coefficients r FResult FTable Probability (p)

(To, °C) a b c
Bud break
5 0.000181 - 0.4676 328.70 0.754 8.58 3.810 0.004
6 0.000126 - 0.3378 246.80 0.709 6.55 3.810 0.011
7 0.000130 - 0.3474 246.50 0.666 5.18 3.810 0.022
8 0.000068 - 0.2039 159.60 0.563 3.02 3.810 0.084
9 0.000034 - 0.1171 102.80 0.465 1.79 3.810 0.205
10 0.000025 - 0.0893 79.30 0.415 1.35 3.810 0.293
Shoot bud
10 - 0.000033 0.0590 - 0.40 0.307 0.68 3.810 0.526
11 - 0.000041 0.0813 - 19.80 0.261 0.47 3.810 0.633
12 - 0.000010 0.0021 26.00 0.415 1.35 3.810 0.292
13 - 0.000014 0.0006 28.50 0.536 2.62 3.810 0.111
14 0.000010 - 0.0527 53.80 0.531 2.56 3.810 0.116
15 0.000012 - 0.0473 41.45 0.474 1.89 3.810 0.190
Leaf and Flowering bud
15 0.000145 - 0.3904 290.60 0.501 2.18 3.810 0.152
16 0.000178 - 0.4782 334.60 0.589 3.46 3.810 0.063
17 0.000112 - 0.3262 240.10 0.633 4.34 3.810 0.036
18 0.000083 - 0.2427 175.10 0.602 3.68 3.810 0.054
Flowering and harvesting
15 0.001226 - 3.2680 2520.00 0.774 9.71 3.810 0.003
16 0.001198 - 3.1630 2375.00 0.756 8.66 3.810 0.004
17 0.001143 - 3.0080 2213.00 0.758 8.74 3.810 0.004
18 0.001012 - 2.6680 1968.00 0.759 8.84 3.810 0.004
19 0.000888 - 2.3820 1734.00 0.752 8.44 3.810 0.004
20 0.000708 - 1.9460 1431.00 0.744 8.06 3.810 0.005
21 0.000550 - 1.5510 1152.00 0.754 8.54 3.810 0.004
22 0.000433 - 1.2380 919.80 0.749 8.29 3.810 0.005
23 0.000448 - 1.2190 854.20 0.762 9.03 3.810 0.003
24 0.000358 - 0.9810 683.50 0.806 12.00 3.810 0.001
25 0.000262 - 0.7303 511.70 0.794 11.05 3.810 0.002
Equations which reflect the relationships elevation may be effective factors but that other
between elevation and the GDD values of oil factors (frost, humidity, rain, fertilization,
rose plant in different phenological periods irrigation, diseases etc.) may also be effective.
have been obtained, which were then examined Therefore, a more exact determination can be
at a statistical significance value of 5% by way made by considering other factors as well in
of correlation analysis (r), F test and probability addition to the factors of temperature and
(p) values. It was determined that temperatures elevation.
of around 8, 9, 10°C suggested for the bud In addition, the changes in GDD values of oil
break period, all temperatures for the shoot bud rose plant at different phenological periods
period and temperatures of around 15, 16, 18°C with the elevations of meteorological stations,
during the leaf and flower bud period, had a have been examined via Duncan test (Table 4).
relationship with temperature which was not Duncan test was used for determining and
statistically significant. In short, it should be classifying the changes between the location
taken into consideration when developing elevations in the study area and the GDD
equations for the growing periods of the oil values for these locations.
rose plant that factors such as temperature and
556
Table 4. The results of Duncan test grouping according to the relationship between GDD values and altitude for oil
rose
Meteorological Phenological Periods

stations Bud break Shoot bud Leaf and Flowering bud Flowering and
harvesting
Aksu 30.40 b 12.40 c 12.90 d 133.00 d
Atabey 30.70 b 16.20 b 38.10 b 228.50 b
Bağkonak 10.30 d 8.60 c 15.20 d 149.70 c
Barla 10.10 d 14.60 b 21.20 c 218.40 b
Eğirdir 29.30 b 16.10 b 28.60 c 231.20 b
Gelendost 25.50 b 2.70 d 8.60 d 188.20 c
Isparta 29.80 b 21.10 a 34.80 b 214.90 b
Kasımlar 14.30 c 14.80 b 17.50 c 206.90 c
Keçiborlu 29.00 b 14.70 b 33.00 b 246.10 b
Kumdanlı 20.80 c 16.10 b 34.30 b 229.00 b
Senirkent 34.70 a 21.40 b 38.60 b 234.40 b
Sütçüler 39.40 a 23.90 a 56.00 a 310.70 a
Şarkıkaraağaç 22.80 b 12.70 c 17.10 d 114.60 d
Uluborlu 32.20 a 21.60 b 41.70 b 199.00 c
Yalvaç 24.40 b 15.30 b 14.70 d 165.50 c
Yenisar-Bademli 4.70 d 3.70 d 4.20 d 104.60 d
According to the Duncan test applied for It has led to an opinion that temperature and
elevation and the GDD values calculated elevation in equations developed for the
according to the optimum temperature demands different growing periods of the oil rose plant,
for different phenological periods of oil rose may be effective factors for determining the
plant (Table 4), it was determined that the study growing areas. However, it was also concluded
area can be classified into 4 different that other factors with impacts on growing
homogeneous growing groups. The groups (frost, humidity, rain, fertilization, irrigation,
were determined as such: 1. (a) group growing diseases etc.) should also be taken into
area: Sütçüler district; 2. (b) group growing consideration.
area: Atabey, Eğirdir, Isparta, Keçiborlu,
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that the Sütçüler district is the most suited date, fermentation duration and Tween 20 treatment
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Atabey, Eğirdir, Isparta, Keçiborlu, Senirkent oil rose (Rosa damascena Mill.). Industrial Crops and
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Gülcülüğü, 125. Yıl anısına Gülbirlik yayınları-1.
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558
IMPROVING BIOLOGICAL AND PHYSIOLOGICAL

PARAMETERS OF NERIUM OLEANDER L. CUTTINGS
BY USING BIOSTIMULATING SUBSTANCES
Diana VÂȘCĂ-ZAMFIR, Elena DELIAN, Ramona COTRUȚ

Abstract
Nerium oleander is a perennial shrub native to the eastern Mediterranean basin and Southeast Asia. As an ornamental
plant grown in pots, it reaches heights of 1-2 m, with the appearance of a bushy shrub, with fragrant flowers, simple or
double, colourful, blooming from spring until the end of autumn. With a very large adaptive potential, the species is one
of the most popular in floral plants, as an indoor plant and where the climate allows, it can be grown on terraces and
balconies, in hedges, alignments etc. The purpose of this study was to evaluate the impact of treatments with various
rhizogenic biostimulators on the rooting potential of different types of Nerium oleander cuttings in greenhouse
conditions. The experiment was performed in the HORTINVEST greenhouses of the USAMV Bucharest and there were
determined the rooting percentage, the development of the root system and the aerial part, as well as some
physiological indicators: photosynthesis, transpiration and stomatal conductance.
Key words: cuttings, rhizogenic biostimulators, physiological indicators.
INTRODUCTION programs (Elloumi et al., 2017). Moreover,

results obtained by Doganlar et al. (2012) and
Nerium oleander L. is an evergreen shrub or Vázquez et al. (2016) also highlights the
small tree (with 2-5 m in height) from the importance of the species as a bio monitoring
Apocynaceae family, cultivated worldwide as tool for airborne metal pollution in urban areas,
an ornamental flowering plant, in different thanks to its resistance to metals and its
geographical and ecological places (Sinha and exclusion capacity.
Biswas, 2016) thanks to an abundant and long- Even if oleander is native to northern Africa
lasting flowering period from summer until and to the eastern Mediterranean region
late autumn (Argiropoulus and Rhizopoulou, (Bailey, 1976), it is naturalized very easily
2013) and due to its moderate hardiness, (Zibbu and Batra, 2010) and grown as pot
tolerance to different soil characteristics and plant, on and around terraces and balconies, in
low nutrients consumption (Kiran and Prasad, hedges and screen plantings (Simion and
2014). Anton, 2009).
The plant value is due not only by its evergreen As Comeaux (1991) noticed, oleander may
leaves, but also by the terminal flowers look like a small tree if the suckers are removed
organized as clusters, with different beautiful and a few stems are kept. Moreover, by
colors, such as white, red, pink, salmon or light growing it in container can be a good choice to
yellow. plant grown even in cooler climates into
Besides the above mentioned utilities, some of greenhouses and conservatories, or as indoor
the secondary metabolites produced by this plant, that can be grown outside during the
plant have also a pharmacological interests summer, as it was practiced for many years.
(Zibbu and Batra, 2010; Chaudhary et al., Due to the great importance of this species and
2015) due to their antibacterial, antimicrobial, increased demands for seedlings, during the
anti-inflammatory, antinociceptive, and time, breeders have had many preoccupation to
antitumor activity (see review Sinha and obtain new cultivars using different plant
Biswas, 2016). In addition, oleander has a propagation techniques such as: generative
promising potential for use in phytoremediation propagation, vegetative propagation by cuttings
559
(softwood or semi-hardwood cuttings in the The substrate for rooting cuttings was perlite,
spring or summer) or in vitro culture (Ochoa et in alveolar plaques.
al., 2003; Simion and Anton, 2009; Vila et al.,
2010; Aryan and Rani, 2016), but are missing Table 1. Experimental distribution
studies on the interaction between the Experimental variants Description
multiplication methods and physiological
performances of the obtained plants. Vm Untreated
As regard as oleander leaves behavior, the V1 Clonex
researchers were concerned to study the V2 Radistim
V3 BioRoot
influence of stress factors such as water stress
(Björkman et al., 1981; Lenzi et al., 2009),
freezing injury (Syros et al., 2005; Miralles- Rooting and Growth Parameters
Crespo et al., 2011), ozone pollution (Lorenzini The following parameters were determined:
et al., 1999) or temperature acclimation percentage of rooting, roots length, shoots
(Badger et al., 1982) on some physiological and length.
biochemical indicators. Leaf gas exchange physiological indicators
To our knowledge, in our country has not until such as photosynthesis rate (An), transpiration
now done research regarding the oleander gas rate (E), and stomatal conductance (gs),
exchange indicators as related to the vegetative respectively have been quantified using the
multiplication techniques. The objectives of the LCIpro + photosynthesis system equipped with
present study were to evaluate the cuttings type a square analysis camera of 6.25 cm-2, between
influence on rooting performance under 7:00 and 10:00 h a.m. All measurements were
different stimulation treatments and to made in four replicates, three times, by
determine some physiological indicators. analyzing the same leaves. The greenhouse
temperature was below 32°C and the light
MATERIALS AND METHODS intensity was maintained between 500-700
μmol m-2 s-1 to avoid the photo inhibition
Plant material process. The results were expressed as: An
Oleander plants (Nerium oleander L.) were (μmol CO2 m-2 s-1), E (mmols H2O m-2 s-1) and
grown in a computer-controlled greenhouse of gs (mol H2O m-2 s-1) and represent the mean
the HORTINVEST Centre at USAMV values for the two cuttings types.
Bucharest, Romania (44º 26' N and 26º 06' E
latitude and longitude, respectively). RESULTS AND DISCUSSIONS
The plants were obtained by vegetative
propagation, using two different type of stem Rooting and Growth
cuttings, the segment ones (4-6 cm long) and The experimental results show that oleander
the peak ones (length of 8-10 cm). cutting rooting was improved by the
After sampling, the cuttings were cut out by stimulating substances.
removing the leaves from the lower node for The effect of the cuttings treatments was
rooting. A number of 12 cuttings of each noticed for both cuttings types, and the medium
category were used for each variant. and hormone applied to the cutting modified all
As rhizogenic biostimulators were used three the studied parameters.
different commercial substances and three The rooting percentage (Figure 1), for the peak
replicates were prepared: shoot cuttings recorded the highest value on V3
- Clonex (a gel, a powerful formula of (100%), followed in decreasing order by V2
hormones, vitamins and minerals); (83.33 %), V1 (75%) and Vm (66.67 %). The
- Radistim (a powder containing non-hazardous order for the other type of cuttings was the
bioactive preparations); same, with the mention that on V1 the value of
- BioRoot (contains vitamins, enzymes, organic the parameter was equal on both types of
acids, humic acids to stimulate root mass cuttings (75%). The rooting percentage for the
growth) (Table1). peak shoot cuttings was higher, with the
mentioned exception of V1.
560
Peak shoot cuttings Peak shoot cuttings
Segment shoot cuttings Segment shoot cuttings 15,1
100,00
16 13,75
100,00 83,33 12,3
14
Rooting percentage (%)
75,00 75,00 80,00 11,1 10,5

66,67 75,00 9,7 10,25
80,00 12
Shoots lenght (cm)

50,00 7,6
60,00 10
8
40,00
6
20,00 4
2
0,00
Vm V1 V2 V3 0
Vm V1 V2 V3
Figure 1. The rooting percentage Figure 3. The shoots length on the new plant
of the cuttings obtained by cuttings
Leaf gas exchange

For the root length, as it is showed in the Photosynthesis rate (Table 2) varied between
Figure 2, the value was highest on the same 1.90 μmol CO2 m-2 s-1 and 3.90 μmol CO2 m-2
type of cuttings (the peak shoot ones), this time s-1and presented higher values in the case of V3,
for all the experimental variants. The order was as compared with the others treated variants
also the same as for the rooting percentage: V3, (V1 and V2), only in the case of the first
followed by V2, V1 and Vm. analysis data. Similarly, transpiration rate (E)
The difference between variants was from had higher values in the case of V3.
21.49% (V3 vs. Vm) to 12.40% (V3 vs. V2) on For stomatal conductance, the values were
the new plants obtained from peak shoot generally low and near to those reported by
cuttings and from 28.45% (V3 vs. Vm) to Delaney (2012) (0.03 mol H2O m-2 s-1 - 0.06
18.10% (V3 vs. V2) in the case of the segment mol H2O m-2 s-1).
shoot cuttings new plants. As regard as oleander multiplication, previous
Peak shoot cuttings studies found that the seeds germination
Segment shoot cuttings percentage varied between 82% and 100%
14 12,1
10,1 10,6
11,6 (corresponding to two determination periods)
12
9,5 8,7 9,5 and the germination time was five days in the
10 8,3
Roots lenght (cm)
both study cases.

8
A better rooting was obtained in the case of tip
6
cuttings on parapet and rooting system for
4
cuttings with temperature control, in peat and
2
perlite (2:1) in 22°C warming culture medium
0
Vm V1 V2 V3
(Simion and Anton, 2009).
As authors emphasized, the best period for
micro propagation was July, on the Murashige
& Skoog medium.
Figure 2. The roots length on the new plant
According to Ochoa et al. (2003) the larger root
growth was obtained by using basal cuttings,
Regarding the result for the third parameter, the
while in the case of the apical cuttings, a longer
shoot length on new plants obtained from
roots number and a higher homogeneity in their
cuttings (Figure 3) it can be seen that the
distribution was noticed.
tendency is preserved: the highest value are on
Micro propagation by using axillary shoot
V3 for the both type of cuttings (15.1/10.5 cm
breaking of wild plants and commercial
peak shoot/segment shoot cuttings), followed
cultivars allowed higher multiplication rates
by V2 (13.75/10.25 cm), V1 (12.3/9.7 cm) and
than the propagation by cuttings, and rooting
finally Vm (11.1/7.6 cm).
and acclimatization did not limited the efficient
production of plants (Vila et al., 2003).
561
Table 2. Leaf gas exchange parameters (mean values ± standard errors)
Parameter Data Control V1 V2 V3
25th May 3.30±0.30 3.00±0.36 3.00±0.15 3.90±0.11
An (μmol CO2 m-2 s-1 ) 11th July 3.62±0.29 3.05±0.25 3.21±0.24 3.89±0.41
12th Sept 2.65±0.66 1.90±0.38 2.51±1.33 2.18±0.10
25th May 0.54±0.07 0.50±0.07 0.50±0.05 0.73±0.04
E (mmol H2O m-2 s -1) 11th July 0.93±0.09 0.90±0.29 1.21±0.09 1.49±0.04
12th Sept 0.55±0.08 0.78±0.25 0.52±0.15 0.62±0.09
gs (mol H2O m-2 s -1) 25th May 0.04±0.008 0.02±0.004 0.03±0.003 0.03±0.002
11th July 0.043±0.004 0.039±0.01 0.046±0.004 0.05±0.004
th
12 Sept 0.028±0.005 0.027±0.01 0.02±0.007 0.03±0.003
Legend: An= Net photosynthesis rate; E= Transpiration rate; gs = Stomatal conductance.
The four studied variants showed some minor hardening. The values obtained for the
differences in leaf gas exchange parameter. photosynthesis rate in the case of the control
Temperature increase determined a higher plants were positive, around 2.21
oleander leaves photosynthesis rate thanks to (photosynthesis higher than respiration) and
improved stability of enzymes involved in negative values in the case of temperature
photosynthesis (Badger et al., 1982). levels under 0°C. Also, in the case of
Transpiration is performed mainly by stomata transpiration, the values were positive (0.32),
complexes and their position, density and while at negative temperatures the values were
opening degree have generally a major impact negative, up to -0.72 at -8°C. Higher
on this process rate, interacting with the photosynthesis rate was reported by Lenzi et al.
stomata conductance. Given that the oleander (2009) (above 8 μmol CO2 m-2 s-1 ) in the case
leaf stomata are located in crypts filled with of fully irrigated plants and in the case of
trichomes, it was expected that water loss to be Angiolo Pucci cv. it was registered the highest
reduced, but studied carried out by Losch et al. carbon dioxide assimilation (An- 10.97 μmol
(1982) emphasize that as compared with other CO2 m-2 s-1), as well as the highest transpiration
species, stomatal crypts were not necessarily rate of 3.24 mmol m-2 s-1 was registered. In the
linked with greater leaf resistance. same time, for this cv. stomatal conductance
According to the results of Roth-Nebelsick et was higher too, 114.72 mmol m-2 s-1.
al. (2009), in the case of water stress situations It was reported a significant decrease of
at the soil level, when the stomata will close to photosynthesis in Nerium oleander in the case
preserve the absorbed water, crypts influence is of ozone treatments and this behavior was
one minor. However, if the growing conditions associated to a partial stomatal closure
are some that allow a higher stomata (Lorenzini et al., 1999). In terms of light
conductance, crypts presence and their effect intensity impact, the fotoinhibition effect on
on reducing water loss are important factors. In oleander chloroplast activity caused by a
such a context, in a previous paper, Gollan et decrease in leaf water potential was lower in
al. (1985) noticed that there was not a critical the case of shade conditions, compared to the
relationship between leaf water potential and plants growth of in full sun (Bjorkman and
leaf conductance, thus, gas exchange decreased Powles, 1981). On the other hand, experiments
was a consequence of soil water content, while performed by Badger et al. (1982) led to the
making reference to abscisic acid hypothesis, conclusion that in the case of oleander,
about the influence on stomatal movement and temperature acclimation it was not required the
indirectly on photosynthesis. According to activation state of Rubisco enzyme. The
Miralles-Crespo et al. (2011), a rapid method to influence of different temperatures was
characterise the impact of freezing injury in explained as follows: at low temperatures it has
oleander is measuring chlorophyll fluorescence. accumulated a higher amount of proteins
Also, Syros et al. (2004) tested the ability of involved in photosynthesis, to ensure higher
six-month-old oleander grown in pot, and rates of photosynthesis and on the other hand,
obtained from propagation by cuttings, to cold leaves grown at higher temperatures have
562
managed to achieve higher photosynthetic rate, the leaves of some shrub, palm and tree species in
due to increased heat tolerance of some urban areas of Adana, Turkey. Chemical Speciation
and Bioavailability, 24 (2), 65-78.
enzymes, which are active in the carbon Elloumi N., Belhaj D., Mseddi S., Zouari M., Abdallah
dioxide reduction cycle (Badger et al., 1982). F.B., Woodward S., Kallel M., 2017. Response of
Nerium oleander to phosphogypsum amendment and
CONCLUSIONS its potential use for phytoremediation. Ecological
Engineering, 99, 164-171.
Kiran C., Prasad D.N., 2014. A Review on: Nerium
The experimental data show that oleander oleander Linn. (Kaner). International Journal of
cutting rooting was improved by the Pharmacognosy and Phytochemical Research, 6 (3),
stimulating substances and the best results were 593-597.
particularly obtained by BioRoot. Gitelson A.A., Buschmann C., Lichtenthaler H.K., 1999.
The effect of the cuttings treatments was The Chlorophyll Fluorescence Ratio F735/F700 as an
Accurate Measure of Chlorophyll Content in Plants
noticed for both cuttings types, and the culture Remote Sens. Enviro, 69, 296-302.
medium and hormone applied to the cutting Gollan T., Turner N.C., Schulze E.D., 1985. The
modified all the biological studied parameters. responses of stomata and leaf gas exchange to vapour
The physiological indicators varied especially pressure deficits and soil water content. Oecologia 65,
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Lenzi A., Pittas L., Martinelli T., Lombardi P., Tesi R.,
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stimulation, in a close interrelation with the cultivars suitable for pot plant production. Scientia
particularities of plant rooting and growth. Horticulturae, 122, 426-431.
Lorenzini G., Nali C., Ligasacchi G., Ambrogi R., 1999.
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and underground organs and its use as a biomonitoring
564
EFFECT OF RHIZOGENIC BIOSTIMULATORS ON ROSMARINUS

OFFICINALIS ROOTED CUTTINGS BIOCHEMICAL COMPOSITION
Diana VÂȘCĂ-ZAMFIR, Daniela BĂLAN, Gabriela LUȚĂ, Evelina GHERGHINA

Abstract
Originally from the Mediterranean area, Rosmarinus officinalis (Fam. Lamiaceae) is a species widespread in most
temperate regions of Europe and America with a significant economic impact due to the trivalent effect (ornamental,
aromatic and medicinal). The purpose of these researches was to evaluate the impact of rhizogenic biostimulator
Clonex-gel treatments on the rooting potential and some biochemical parameters of different types of cuttings in
greenhouse conditions. The experiments carried out in the HORTINVEST greenhouses of USAMV Bucharest aimed to
evaluate the rooting percentage, the development of the root system and the aerial part after rooting. Also, some
biochemical parameters (assimilatory pigments and total soluble sugars content) were analyzed to assess the potential effect
of the substances used to stimulate rooting on the biochemical composition of leaves formed on cuttings. The researches
performed indicated that maximum percentage of rooting cuttings was determined in treated variants. The stimulation of the
roots growth determined also an increased accumulation of assimilatory pigments and soluble glucids in the leaves.
Key words: carotenoids, chlorophyll, cuttings, rooting potential, rosemary.
INTRODUCTION Chakravarti et al., 2005) and also it was used as

a natural antimicrobial and antifungal against
Rosemary (Rosmarinus officinalis) belongs to E. coli, Pseudomonas, Aspergillus,
the family Lamiaceae (Labiatae) and is native Staphylococcus or an insecticide (Duke, 2001).
to southern Europe and the Mediterranean area. Besides this, it was reported that the leaves
Nowadays, rosemary is grown in almost all have high antioxidant activity attributed to its
countries around the Mediterranean but also in phenolic compounds (Singh and Guleria, 2013)
England, USA and Mexico. In the countries of therefore rosemary extracts have been widely
the Mediterranean, it is a popular spice espe- used as a preservative in the food industry
cially in Italy and France, and less in Greece (Bozin et al., 2007).
and Spain. One hundred grams of fresh rosemary leaves
If for centuries it has been used as a herbal contain approx. 67.77 g of water, 3.31 g of
aroma, natural medicine and a basic ingredient protein, 20.70 g of carbohydrates, and 14.1 g of
in the perfume industry today rosemary can be fiber. Among the minerals are Ca (317 mg),
defined as a trivalent species, in addition to its Mg (91 mg), P (66 mg), K (668 mg), Na (26
medicinal uses. So, it is cultivated as an mg), Fe (6.65 mg), Zn (0.93 mg). Vitamins are
aromatic plant in culinary art and also as a mainly B6 (0.336 mg), Thiamin (0.036 mg), C
decorative plant for fragrances, protection from (21.8 mg), Riboflavin (0.152 mg) and others
pests and gardening. (USDA, 2016).
Rosemary has been longtime considered an Oils and vinegar are flavored with rosemary
important plant for its essential oil used not branches. It can be found in tea blends (Tisane)
only in perfumes and as food spice (Moss et al., or spices (herbs of Provence). In combination
2003), but also in medicine (Miguel et al., with most Mediterranean herbs (oregano, basil
2007). etc.) it tastes good but can also be associated
The whole plant was reported to possess seve- with garlic and thyme. The rosemary leaves
ral medicinal properties like cardiac, antiseptic, from the culinary point of view can be used
antispasmodic, carminative, cholagogue, fresh, but also dry (the aroma is not lost, even if
nervine spasmodic, astringent, stomachic, and the plant is treated for a long time).
tonic (Grieve, 1984; Polunin and Huxley, 1987;
565
Pleasure-smelling foliage shrubs are appealing Long stems from selected mother plants were
in the garden, especially for visually impaired harvested. Two types of stem cuttings were
people. Sometimes the scent feels only in the made: of the segment (4-5 cm long) and peak
vicinity of the bushes, sometimes it has to (length 5-6 cm).
touch or crush the leaves between the fingers to After sampling the cuttings were cut out by
release the perfume (Noordhuis Klaas T., removing the leaves from the lower node for
2008). rooting. A number of 15 cuttings of each
The propagation by cuttings obtained from the category were used for each variant.
stem is a frequently used way of vegetative The substrate for rooting cuttings was river
multiplication of many ornamental plant sand in alveolar plaques.
species. Cutting is well-known as common and Experimental variants were established: 50% of
relatively cheap method because it overcomes the cuttings were subjected to a treatment by
the difficulties of multiplication by plant seeds immersing the base in rooting stimulators
(Elhaak et al., 2015). (Clonex-gel, a powerful formula with a 0,3%
It is well known that the plant-produced concentration of 4-indol-3-yl-butyric acid,
phytohormones like auxins play a key role in containing also vitamins and minerals). As
stimulating the adventitious root development control variants there were V2 and V4 (Table 1).
of stem cuttings and the roots’ branching.
Treating cuttings with auxins aims to increase Table 1. Experimental variants
the percentage of rooting, root initiation, Number
number and uniformity of rooting (Elhaak et Variants Description
of plants
al., 2015). Besides these, cytokinin and gibbe- Shoot peak cuttings treated
V1 15
rellins also contribute to the intensification of with Clonex
metabolic processes, cell division or growth in Shoot peak cuttings without
V2 15
length. Associating such compounds in rooting Clonex
Shoot segment cuttings
substrates allows the increase of the rooted V3
treated with Clonex
15
percentage of the cuttings and in the rooting of Shoot segment cuttings
V4 15
cuttings (Georgescu et al., 2012). without Clonex
Previous study related that rosemary has a good
rooting ability and the effect of the root-
Throughout the period of rooting there were
forming solution (standard macrosalt
applied specific works complex care for
formulation) induces better results, both in
directing microclimate factors.
terms of the rooting percentage and the
After rooting were made observations and mea-
qualitative features of the new root system
surements on rooted cuttings, on the percentage
(Talia et al., 2004).
of rooting, root length, shoot length.
The purpose of these researches was to evaluate
For the rooting percent comparison between
the impact of the treatment with rhizogenic
variants, it was used the Fisher exact test and for
stimulator on the rooting potential and on some
comparison between variants for shoot length and
biochemical parameters in different types of
root length it was used the Student test.
cuttings in greenhouse conditions.
The biochemical parameters were analyzed
The rooting percentage, the development of the
using proper methods:
root system and the aerial part after rooting
Determinations of the assimilatory pigments
together with the content of assimilatory
content in the active leaves: chlorophyll and
pigments and sugars were analyzed to assess
carotenoid pigments were extracted in 80%
the effect of this treatment.
acetone and the absorbance of the extract was
measured at three wavelengths (663 nm, 647
nm and 480 nm) with a UV/Visible
ThermoSpectronic Helios spectrophotometer.
The experiment was conducted in greenhouses
The results were calculated using the extinction
at the Hortinvest Center of the USAMV
coefficients and equations described by
Bucharest (44º 26' N and 26º 06' E latitude and
Schopfer (1989) and were expressed in mg/g
longitude, respectively).
fresh weight.
566
Determination of total soluble sugars was nonsignificant differences between V1 and V3
performed according to the Nelson-Somogyi (df = 28, p = 0.10> 0.05).
method (Iordachescu, 1988; Somogyi, 1952). Similar results have been obtained when
For determination of the total soluble glucid comparing means between variants for root
non-reducing glucids were first transformed in length.
reducing glucids by hydrolysis with hydro- Roots system is significantly involved in plant
chloric acid. The reducing glucids when heated growth due to their role in water and nutrient
with alkaline copper tartrate reduce the copper uptake, therefore maintaining an active roots
from the cupric to cuprous state and thus growth is essential for plant development and
cuprous oxide is formed. When the cuprous productivity.
oxide is treated with arsenomolybdic acid, the It is expected that the bioregulators treatment to
reduction of molybdic acid to molybdenum maintain viability and functionality of the roots
blue takes place. The measurements of system during the growth period.
absorbance were achieved at 540 nm with a Study of biochemical parameters in the
UV/Visible ThermoSpectronic Helios rosemary cuttings in all experimental variants
spectrophotometer. The results were expressed Assimilatory pigments
in mg/g fresh weight. Chlorophylls a and b represent the major
photosynthetic pigments in plants, playing an
RESULTS AND DISCUSSIONS important role in the photochemical reactions
of photosynthesis (Taiz and Zeiger, 2009),
The rhizogenesis process at Rosmarinus while carotenoids are considered as accessory
officinalis cuttings has been carried out quickly, components providing photoprotection (Torres-
so that after approximately 30 days since the Netto et al., 2005; Simkin et al. 2008).
placement of the cuttings on the rooting Previous studies have been reported increased
medium, they have formed roots. amounts of assimilatory pigments under
The number of new plants (rooted cuttings) applications of some growth biostimulators in
obtained differs depending on the variant. Also, tomato leaves (Khan et al., 2009), Physocarpus
as the rooting process takes place, there are opulifolius stem cuttings (Pacholczak et al.,
new vegetative growths of the cuttings' 2016), dogwood cuttings (Pacholczak et al.
terminal bud. 2012), wheat (Vician and Kováčik, 2013) or
Study of biometric parameters in the rosemary tomato (Mikiciuk and Dobromilska, 2014). On
cuttings in all experimental variants the contrary, Tsai and Arteca (1985) noticed
The data on the number of cuttings obtained that applications of gibberellic acid decreased
after rooting were summarized in Table 2. the chlorophyll content in barley, squash,
pepper, sorghum, pigweed and kochia, while
Table 2. The parameters of new plants obtained by in oat, wheat, mung bean, corn, millet and
cuttings
gomphrena it remained unchanged. Also,
Variants Rooting Shoot Root Vasca-Zamfir et al. (2012) found that treatment
percent (%) length (cm) length (cm) of the plants with root stimulators like Clonex,
V1 93.33 10.84 13.22
V2 80.00 8.60 12.10
Coralite and Radistim did not significantly
V3 86.66 9.43 13.14 influence the content in assimilatory pigments
V4 66.66 6.90 9.90 in the leaves of Pelargonium peltatum, while
Elhaak et al. (2015) made similar observation
Thus, it can be seen that the values for the main regarding the treatment with IBA (indole-3-
parameters are superior in the variants treated butyric acid) of rosemary.
with Clonex (V1 and V3). Regarding the values The researches made on rosemary cuttings
for each type of seedlings, it is noticeable that showed that generally the biostimulator
the peak ones (V1) gave better results. treatment influenced the contents of organic
According to the Fisher exact test results, there compounds. The contents of total chlorophyll
is no significant difference for the rooting in the cuttings treated with Clonex (V1 and V3)
percent between variants. Comparing the shoot increased relative to the control plants (V2 and
length mean with the Student test, there was V4) in both experimental variants (Table 3).
567
However, the cuttings provided from peak However, the chlorophyll (a+b)/carotenoids
registered higher values of the chlorophyll total ratio increased in the variants treated with the
content by comparison with shoot segment biostimulator due to an increase of
cuttings. chlorophylls a and b rather than of carotenoids.
Increased chlorophyll (a+b)/carotenoids ratio
Table 3. The impact of rhizogenic biostimulator found also Elhaak et al. (2015) in rosemary
treatments on total chlorophyll and glucids content cuttings after soaking for one and three hours in
in rosemary cuttings
IBA but the prolonged time of soaking (six
Variants Total chlorophyll Total hours) decreased the ratio probably because of
(mg/g f.w.) soluble glucids
(mg/g f.w.)
an inhibition in the metabolism of chlorophylls
a and b. Sosnowski et al. (2016) reported after
V1 2.03±0.04 1.32±0.03
treating Medicago plants with different growth
V2 1.41±0.03 0.92±0.02
V3 1.48±0.05 0.82±0.05 regulators that the highest concentration of
V4 1.36±0.03 0.56±0.03 carotenoids was found in the Medicago leaves
sprayed twice with cytokinin while auxin
It can be noted higher amounts of both decreased their content and a mixture of auxin
chlorophylls a and b in the cuttings which and cytokinin raised the ratio of total
received stimulatory treatments, while the ratio chlorophyll content to carotenoids.
chlorophyll a/b was not significantly influenced Total soluble glucids
by the Clonex application (about 2.8 in the Sugars are considered important metabolites, is
cuttings from the peak, respectively 2.9 in the not only the first organic complex compounds
cuttings from segment whether treated or not) formed in the leaves as a result of
(Table 4). photosynthesis, but also a major respiratory
substrate. Also sugars are involved in plant
Table 4. Ratio of assimilatory pigments protection against wound and infections, as
in the variants well as in cell detoxification (Kaur et al., 2000).
Variants Chlorophyll Total chlorophyll / The results of the present researches indicated
a/b carotenoids that the stimulation of the roots growth
V1 2.82 23.36 determined an increased accumulation of
V2 2.88 20.67 glucids in the leaves of rosemary cuttings
V3 2.97 21.45 (Table 3). Both experimental variants showed
V4 2.95 20.90 higher amounts of total soluble glucids in the
stem cuttings treated with Clonex comparing to
Regarding the carotenoids content, the amounts the control cuttings: about 1.43 times higher in
determined showed a slight variation in the the shoot peak cuttings, respectively 1.46 times
analyzed rosemary leaves in all experimental higher in the segment cuttings.
variants (Figure 1) indicating that Clonex Scientific literature noted that higher
treatment not affected this parameter. photosynthetic pigment concentrations in
cuttings treated with growth regulators like
1,6 auxin significantly contribute to increased
1,4 photosynthetic activity and enhanced produc-
1,2 tion of photoassimilates. Consequently, higher
1 glucids levels during rooting are positively
(mg/g)
0,8 correlated with increased root formation and

0,6 more nutrients uptake of cuttings (Krajnc et al.,
0,4 2013).
0,2 Also, Pacholczak et al. (2016) found that the
0 treatments with Goteo biostimulator increased
V1 V2 V3 V4 total soluble and reducing sugar contents in
Chlorophyll a Chlorophyll b Carotenoids Physocarpus opulifolius stem cuttings. Similar
Figure 1. Accumulation of assimilatory pigments results obtained Rathore et al. (2009) in
in the experimental variants Glycine max treated with a preparation based
568
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570
COPPER EFFECT ON SEED GERMINATION AND PLANT SPROUTING

OF ALYSSUM MURALE SPECIES
Maria BRÎNZĂ, Elena Liliana CHELARIU, Andreea AIORDĂCHIOAIEI, Lucia DRAGHIA
,,Ion Ionescu de la Brad” University of Agricultural Sciences and Veterinary Medicine of Iaşi,
3 Mihail Sadoveanu Alley, 700490, Iaşi, Romania
Abstract
The studies aimed at establishing the level of copper concentration in the soil, which Alyssum murale species can tolerate
without significantly affecting the percentage of germination and plant sprouting. The experience was conducted in 4
variants with 3 replicates, each replicate having 100 seeds. The humidity of the substrate for sowing was performed with
water for the control sample (C) and with CuSO4 solutions for the other variants (it was calculated the required amount of
CuSO4 to obtain a contamination with Cu of 20 ppm, 100 ppm, 200 ppm and, respectively, 500 ppm). The influence of
copper on seed germination was assessed by the percentage of plant sprouting, germination rate, seedling rate and
velocity. The significant decrease in the percentage of normal sprouts and the increase of percentage of dead seeds in
variants V3 and V4 suggest the level of supportability of these species. The increase of copper concentration in the
germination substrate caused the decrease of the seedling rate and of the velocity of seedling.
Key words: Alyssum murale, copper, germination, sprouting percentage.
INTRODUCTION Copper (Cu) represents one of the first metals

that were extracted and used by humans in
Pollution has become a major concern in all different activities, thus providing a major
industrialized countries, at a global level, since contribution to the evolution of human society
every year, millions of tons of toxic pollutants since ancient times. Research done on the
originating from natural sources, but more toxicity of this particular metal has highlighted
particularly from anthropogenic sources, are a series of anthropogenic sources of emission
released into the environment. As a result, the of copper in the environment, such as: copper
research carried out over the years has extraction and processing, the agriculture, the
highlighted the favourable effect that some electric power industry, the plastics industry,
plants have on the depollution of the wastewater sludge (animal farms) and
environment by absorbing the pollutants and steelworks.
accumulating them in different parts of the In low quantity, copper constitutes the essential
plant (Wenzel and Jockwer, 1999; Clemens, element for a proper development in plants,
2001; Wierzbicka, 2002; Baranowska-Morek, playing an important role in photosynthesis,
2003). Phytoremediation is the use of respiration, lignification and growth, as well as
spontaneous and/or cultivated plant species in being a constituent of proteins and enzymes.
order to extract, stabilize and/or neutralize In excess, copper can become extremely toxic,
pollutants in soils. Over time, research in affecting seed germination and causing a weak
phytoremediation showed favorable effect of root and stem growth, as well as leaf chlorosis
certain plants species, including ornamental and necrosis (Yruela, 2005; Xu et al., 2005;
plants, indicating pollution reduction by Muhammad A. et al., 2015).
absorbing pollutants by means of plant roots Various studies have reported the germination
and accumulating them inside the plant. Lately, test as being a basic process of determining the
phytoremediation has become a well-studied toxicity effects of Cu in numerous plant
scientific subject, which has led to an species. The reduction of the germination
interdisciplinary cooperation between percentage as a result of the presence or
biochemists, biologists, soil chemists, increased concentrations of copper in the
agronomists and environmental engineers (Salt germination environment was highlighted in
et al., 1995). different species: Sunflower (Pena et al., 2011;
571
Boroş et al., 2015); Vicia sativa L. (Muccifora RESULTS AND DISCUSSIONS
and Bellani, 2013); Minuartia hirsuta, Silene
compacta, Alyssum montanum (Ouzounidou et The observations regarding the influence of
al., 1994); Phaseolus vulgaris L. (Ashagre et copper on the Alyssum murale seed germi-
al., 2013); Oryza sativa L. (Ahsan et. al., 2007; nation highlight a decrease of the germination
Mahmood et al., 2007) percentage occurring at the same time as an
The results of the studies suggest that Cu increase in the copper concentration.
toxicity in relation to the seed germination By comparing the results obtained in all experi-
process in different plants can present a mental variants, the influence of copper pol-
remarkable variability of tolerance, both within lution on Alyssum murale seed germination is
the genus as well as between different species. highlighted by the more accentuated decrease
of the germination percentage. The highest
MATERIALS AND METHODS germination percentage was obtained in the
control variant (93%), and the lowest
For the experiment, the biological material used germination percentage was obtained by the V4
was represented by Alyssum murale seeds taken variant (64%) (Table 1).
from the collection of the Faculty’s Floriculture Compared with the control variant, for all the
department, a species that is conserved ex-situ. other experimental variants, there was a
In the experiment were used containers of 2 kg decrease of the germination percentage obser-
capacity, containing the substrate made of two ved, the highest values being recorded for the
parts peat and one part gardening soil, in which V3 variants, of 20%, and V4 variants, of 29%.
100 seeds were sown per container, three The determinations regarding the seed
replicates being done for each experimental germination period have revealed that the
variant. Alyssum murale seed germination was
Watering was realised with tap water for influenced by the concentration of copper.
control sample, and for the other three samples,
with solutions of different CuSO4 concen- Table 1. Influence of copper on the seed germination of
the Alyssum murale species
trations.
The necessary quantity of CuSO4 was Germination
End of
Total
calculated, in order to obtain certain copper Variant Sowing date
onset date
germination
date
germination*
concentrations in the substrate, that represented
the maximum admitted limit of Cu - 20 ppm C 19.03.2014 22.03.2014 26.03. 2014 93
(V1); the warning threshold - 100 ppm, (V2); V1 19.03.2014 22.03.2014 27.03. 2014 88
the intervention threshold - 200 ppm, (V3); V2 19.03.2014 25.03.2014 02.04. 2014 80
exceeding the intervention threshold - 500 ppm V3 19.03.2014 24.03.2014 03.04. 2014 73
(V4). V4 19.03.2014 29.03.2014 09.04. 2014 64
During the experiment, the temperature of the *The values represent % of normal sprouts
vegetation room was constantly maintained at
22 ± 1°C for 8 hours, and at 24 ± 1°C for 16 The seed germination began after 3 days in the
hours. The seed germination was done under case of the control variant and V1, the increase
conditions of relative humidity of of the copper concentration determining a
approximately 80% during the first 3 days and bigger delay of the onset of germination, the
of 87% until the end of the germination period, biggest delays being recorded in the case of the
at a level of light intensity over 8000 lux. V3 variant (4 days, compared to the control
During the entire germination period, daily variant) and V4 variant (4 days, compared to
determinations and observations were made the control variant) (Table 1).
regarding the germination percentage, the Regarding the influence of copper on the
number of normal sprouts, the number of germination period (from the onset until the
abnormal sprouts, the number of dead seeds, end), there is a bigger delay observed in the
the velocity of seedling and the coefficient of case of the variants that were exposed to higher
velocity (Kotowski, 1962). doses of copper (Table 1).
Figure 1 illustrates that, under conditions of
copper contamination, the number of days from
572
the onset of germination of the Alyssum murale
seeds, up until its end, ranged between 4 days
in the case of the control variant and 12 days
in V4.
Figure 2. Ratio of abnormal sprouts and dead seeds (%)
Compared to the control variant, the number of

abnormal sprouts was much higher in the case
of the contaminated variants, the most evident
Figure 1. Seed germination duration under conditions of
increases in numbers being obtained for the V4
copper contamination (number of days) variant, by 17%, followed by the V3 variant,
with an increase of 11%.
The period of time from sowing until the onset By comparing the number of dead seeds
of germination was of 3 days, in the case of the obtained in the case of the contaminated
seeds originating from the control variant and variants with the one obtained by the control
V1 variant, of 6 days for the V2 variant and of 8 variant, the highest increases in numbers can be
days for the V3 variant. observed in the case of the variants that had
Regarding the V4 variant, the results regarding exhibited the highest number of abnormal
the seed germination were obtained during a sprouts as well (12% for V4 and 9% for V3).
period of 12 days. The results regarding the seedling growth rate
Compared to the control variant, the biggest (Table 2) indicate the tendency of a decrease in
delay regarding the number of days from the germination percentages, in the case of the
onset to the end of germination, was observed variants that show a metal content that repre-
in the case of the V4 variant (8 days), followed sents the intervention threshold and a
by the V3 variant (4 days). concentration over the intervention threshold
In the case of the variants contaminated with (V3 and V4).
copper, there may be observed an increase in Table 2. General characterization of the seedling of the
the number of abnormal sprouts, as well as the Alyssum murale species under exposure to different
ratio of dead seeds (Figure 2). doses of copper
In the case of the Alyssum murale species, Velocity of Velocity
Seedling Seedling
under conditions of copper contamination, Variant
(%) duration
seedling coefficient
there may be observed an increase in the (%) (%)
C 82.5 3 27.5 4.71
number of abnormal sprouts, for the variants
that were exposed to the highest doses of Cu. V1 77.5 3 25.8 3.88
The observations made also highlight a change V2 67.5 6 11.3 2.08
in the number of abnormal sprouts, the V3 20.0 7 2.9 1.33
differences compared to the control variant V4 16.0 10 1.6 0.73
being highly significant.
In the case of the control variant, out of the The long time span necessary from sowing until
total number of seeds used in determining the the onset of seedling, correlated with the small
germination, 93% have generated normal number of seedlings obtained in the first days, in
sprouts, 4% have generated abnormal sprouts the case of the variants exposed to high doses of
and 3% were dead seeds. The increase in the copper, determine reduced values of velocity
copper concentration has determined an and coefficient of velocity.
accentuated increase in the percentage of The velocity (Table 2) registered the lowest
abnormal sprouts and dead seeds (Figure 2).
573
value in the case of the V4 variant (0.73%) and annuus). Studia UBB Ambientum, LX, 1-2, 2015,
the highest value in the case of the control 23-30.
Clemens S., 2001. Molecular mechanisms of plant metal
variant (4.71%). A similar situation may be tolerance and homeostasis. Planta 212: 475-486.
observed in the case of the coefficient of the Draghia L., Chelariu E.l., Zaharia A., 2011. Aspects
velocity of seedling, where the lowest value is regarding the production of planting material at some
recorded in the case of the V4 variant (1.6%) ornamental species from spontaneous flora. Bulletin
and the highest value in the case of the control of University of Agricultural Sciences and Veterinary
Medicine Cluj-Napoca. Horticulture. Print ISSN
variant (27.5%). 1843-5246; Electronic ISSN 1843-5386, vol 68, no.
1, 332-337.
CONCLUSIONS Gang A., Vyas A., Vyas H., 2013. Toxic effect of heavy
metals on germination and seedling growth of wheat.
The influence of copper pollution on the Jenviron Res Develop, 8: 206.
Alyssum murale seed germination has been Kotowski F., 1962. Temperature relations to germination
highlighted by the more accentuated decrease of vegetable seeds. Proc. Amer. Soc. Hort. Sci. 23:
of the germination percentage in the case of the 176-177.
Mahmood T., Islam K.R., Muhammad S., 2007. Toxic
variants with contaminated substrate, the effects of heavymetals on early growth and tolerance
highest values being recorded in the V3 variant, of cereal crops. Pak J Bot 39: 451.
of 20% and the V4 variant, of 29%. Muccifora S., Bellani L.M., 2013. Effects of copper on
Higher concentrations of copper have deter- germination and reserve mobilization in Vicia sativa
mined a bigger delay regarding the number of L. seeds. Environ Pollut 179: 68.
Muhammad A., Shafaqat A., Muhammad R.,
days from the onset until the end of germi- Muhammad I., Farhat A., Mujahid F., Muhammad Z.,
nation, which was more evident in the case of Muhammad K.I., Saima A.B., 2015. The effect of
the V4 variant (8 days) and V3 variant (4 days). excess copper on growth and physiology of important
The development of the sprouts was affected by food crops: a review. Environmental Science and
the increase in the concentration of copper, Pollution Research. Volume 22, Issue 11, 8148-8162.
Ouzounidou G., 1995. Effect of copper on germination
resulting in a higher number of abnormal and seedling growth of Minuartia, Silene, Alyssum
sprouts and dead seeds in the case of V4 and V3 and Thlaspi. Biologia Plantarum. 37: 411.
variants. Pena L.B., Azpilicueta C.E, Gallego S.M., 2011.
The long time span necessary from sowing Sunflower cotyledons cope with copper stress by
until the onset of seedling, correlated with the inducing catalase subunits less sensitive to oxidation.
J Trace Elem Med Biol. 25: 125-9.
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days, in the case of the variants exposed to high B., Chet I. et al., 1995. Phytoremediation: A novel
doses of copper, determine reduced values of strategy for the removal of toxic metals from the
velocity and coefficient of velocity. The re- environment using plants. Nature Biotechnology, 13,
search conducted targeted the practical check pp. 468.
Singh D., Nath K., Sharma Y.K., 2007. Response of
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Alyssum murale. peroxidase activity of Mung bean (Vigna radiata). Int
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(Phaseolus vulgaris L.) cultivars as influenced by Biologica Cracoviensia Series Botanica 44: 7-19.
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Baranowska-Morek A., 2003. Ros´linne mechanizmy Wang Y.L., 2005. Effects of soil copper concen-
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germination and growth of sunflower (Helianthus 17: 145-156.
574
DECODING THE SIGNIFICANCES OF BUILT LANDSCAPE PROFILES
Doina Mira DASCĂLU, Mirela COJOCARIU
,,Ion Ionescu de la Brad” University of Agricultural Sciences and Veterinary Medicine of Iași,
3 Mihail Sadoveanu Alley, Iaşi, 700490, Romania
Abstract
Perceived from a distance, any built landscape emits a specific energy personified by the silhouettes of its profiled
buildings on the background of the environment. Overall images of human settlements have appeared in medieval
artwork, being further perfected. These representations had a symbolic value, contributing to strengthening the respect
for tradition and the local history. Towards the middle of the 20th century, architect G.M. Cantacuzino created several
sketches of built landscape profiles, both historic and modern. The theme of this paper is based on these sketches. The
research aims to argue that by elaborating certain built landscape profiles and then by decoding, analysing and by
comparative assessment of the meanings transmitted by silhouettes, valuable information can be obtained about the
essence, identity, personality and specificity of those human settlements. Research has been made on the basis of
selected sketches of Cantacuzino and on the basis of the creation of original profile sketches of historical settlements.
The arguments in this paper on decoding the profiles of built landscape contribute to highlighting a valuable tool useful
for landscape researches and urbanism studies.
Key words: decoding, features, landscape, profiles, settlements.
INTRODUCTION evolution: the natural environment, the built

environment, and the socio-economic-cultural
It is important to understand that human environment (Dascălu and Cojocariu, 2016).
settlements can be considered as "the most The idea of this work has its roots in the
conclusive possession" of a natural space and historical and modern built profiles, drawn by
its transformation according to the aspirations the architect George Matei Cantacuzino in his
and needs of the respective human society youth. Sketching the silhouettes, he tried to
(Gheorghiu, 2002). characterize the historical personality of human
Analysing the history of an epoch and its settlements he visited and loved. His creations
influence on the evolution of urban tissue and were impregnated with great passion for the
the built framework, we observe that some built landscape, nature playing a very important
important political-social-economic-religious- role in his life (Dascălu, 2017).
cultural processes in the local history are In one of the volumes of Art and Criticism
imprinted in the volumetric silhouette of human Dictionaries entitled Symmetry, written
settlements. between 1939 and 1946, the architect manages
Perceived from a distance, any built landscape to define the profile in general, either in art or
emits a special message, personified by the in architecture, urbanism or landscaping: „The
building's overall silhouettes. profile is the line that includes the appearance
Based on judicious documentation, studying of an object (or a settlement) ... It is a line that
stamps, manuscripts, frescoes, paintings, expresses its self schematically ... The profile
photographs and old maps, we can sketch or suggests forms without expressing them ... and
draw the outline of built landscape profile, tends to ideogram” (Vasilescu, 1993).
extracting and deciphering certain So at first glance, a built landscape profile
characteristic features, essential for the transmits a synthetic idea about that human
respective human settlement, such as identity, settlement and its inhabitants.
personality, specificity and value. In this context the paper aims to demonstrate
We can also discover synthetic features of the that certain features specific to historic cities,
environmental elements involved in historical such as defensive or aggressive aspect,
575
dominant or peaceful ambience etc., can be The demonstration continued with the
clearly decoded by drawing the general profile researches of two case studies of historical
of a built landscape, outlined on the sky and on cities from France, chosen as a result of
the background of the natural setting. documentary research: the city of Carcassone,
In order to extract more detailed information which has some resemblance to San Gimignano
from the silhouette of a profile, it is necessary and the small settlement of Castelnau-le-Lez.
to study the history of the places but also to For these settlements were studied historical
analyse the environmental elements involved in frescoes, paintings and engravings representing
the evolution of those human settlements. eloquently the built frame and their natural
Besides all this, on the basis of written landscape. The old images have been checked
documents and old images, it is necessary to and compared with current photos and images.
create several profiles on successive periods. In order to complete the decoding of these
The information extracted through sketching settlements personality it was necessary a
and through correlated and comparative documentary on the history of places and on
research can finally reveal unexpected features environmental elements.
that could be necessary in the contemporary The elaboration of original profiles, based on
studies of urban and landscape development of selected paintings and photographs, was the
localities and territories. most important part of the research,
The ideas and the research in this work are contributing to the deciphering of the
original. No other similar research is known. personality of the studied settlements.
Different studies about city abstraction as icons
or lines and its skylines have been aimed at RESULTS AND DISCUSSIONS
either clarifying certain aspects of aesthetic or
psychological perception of the urban or The first analyses were made on the basis of
natural landscape, or pursuing certain socio- selected silhouettes from G.M. Cantacuzino's
spatial aspects. drawings and on the basis of the elaboration of
Of these studies we mention Gassner's (Gassner, historical settlement profiles, after the research
2009) specifying the difference between his on the history of the places and on the
study and ours. Gassner-type studies and environmental elements.
sketches aim to improve the city's current Regarding deciphering the meanings of the
silhouettes, impacting cognitively and socially. lines and forms, there is a wide range of
We underline that our research aims to provide aesthetic studies correlated with the psycho-
a tool useful for deciphering the study of logical ones. According to these studies, the
historical evolution of human settlements, sharp, agitated, angular lines can be the result
correlating the local history, the natural chosen of either conflicting states or dynamic
location and the historical silhouettes offered tendencies (from natural, artificial or human
by different authors. environment), while the sinuous, with quiet
curves unfolded suggests situations of stability
MATERIALS AND METHODS and peaceful development (Reid, 2007).
There were chosen two silhouettes for which
Two profiles outlined by G.M. Cantacuzino for the architect Cantacuzino finds similarities in
the cities of San Gimignano and New York- terms of styling of the profile line and the size
Manhattan were selected for analysis. scale of the constructions: San Gimignano from
The analysis investigate the correlation Italy (Figure 1) and Manhattan the heart of
between the historical truth based on the New York USA (Figure 2).
documentation and the profiles drawn by In the dictionary Symmetry - Books and
Cantacuzino. Criticism Cantacuzino draws attention that
Based on the documentary material, the reality „San Gimignano preceded by several centuries
and the veracity of the synthetic features the American ambitions... By comparing the
declared by the architect and decipherable in two profiles, we can be convince that it is both
his drawings, were analyzed and substantiated ambition and pride” (Vasilescu, 1993).
for these profiles.
576
Analyzing the representations of San
Gimignano over time, we find that
characteristic features such as massive and
ascending volumetric trends have been
perceived and presented similarly from
Renaissance time to the 20th century, indicating
that the city has always developed in the same
architectural and constructive manner. The
Figure 1. The profile of San Gimignano city, sketched by
G.M. Cantacuzino following images were selected and compared:
(source: processed from Vasilescu, 1993) the frescoes of the saints Gimignano and Fina,
who both hold and protect the town of San
Gimignano in their arms (exhibited at
Pinacoteca of San Gimignano Palazzo
Comunale); a Benozzo Gozzoli's Renaissance
fresco (exhibited at the apse of the Sant
Agostino-San Gimignano chapel); a picture in
the Cubism manner of 1912 by Pyotr
Konchalovsky (exhibited at State Russian
Museum of Saint Petersburg).
Figure 2. The profile of New York City, As mentioned above, the fortified volumetric
sketched by G.M. Cantacuzino expression of the city could suggest a defensive
(source: processed from Vasilescu, 1993) attitude. But, investigating the history of the
city we discover that during the medieval
San Gimignano is a medieval Italian town period there was a rivalry between two noble
whose historic nucleus is protected by families that dominated economically the city.
UNESCO. The purpose of the documentation This conflict has generated the competitive
was to check why San Gimignano might have a construction of buildings with higher towers,
proud and high-aimed profile. more and more grandiose, in the egotic attempt
A first observation, extracted from the history to leave a trace in the history of the city.
of architecture and urbanism, is the fact that, in In connection with Cantacuzino's comparison
general, ascending architectural volumes of San Gimignano and New York, we
suggests ambition and pride (Gheorghiu, 2009). examined many artistic representations of the
Despite this fact, history demonstrates often two cities and selected two artists whose works,
that ascending features are chosen to be in Expressionist manner, crystallize the essence
expressed because impressive and massive of the profiles: San Gimignano by M.C. Escher
volumes can demoralize any attacks of enemies painted in 1922 and Manhattan by Howard
(Gheorghiu, 2009). Cook painted in 1930. The comparison of their
Related to these two possibilities, studying the drawings is eloquent to distinguish the same
conditions for choosing the location of the early ascending traits in which the message of
urban core, we noted the special defense ambition and pride is transmitted.
possibilities were offered by the local relief, Research has continued with the study of New
proving their importance. The city is located on York history, which has highlighted that over
one of the hills of Tuscany, therefore the site time the city has tended to play an increasingly
have a good natural defense. As a result of the important economic role in the USA.
research, we discovered a painting of George Consequently, towards the end of the
Howard at the end of the 19th century, clearly nineteenth century, the first skyscrapers in New
distinguishing the territorial strategic location, York appeared to be the tallest, in competition
the city's silhouette being legible on the with Chicago buildings for example.
impressive hillside (George Howard’s The tendency to build upright has increased
watercolour „La citta della belle torre-San over time, making Manhattan the heart of New
Gimignano”, 1877, exhibited at London, York and the symbol of USA economic power.
Grosvenor Gallery). Unfortunately, this trend is tragically linked to
577
the 2011 attacks that aimed at destroying the represented over time, from the 13th century to
volumetric symbols of New York at the time - the 19th century, with small variations due to
the twin towers of the World Trade Center. the construction of new buildings, but raised in
In conclusion, these researchs on the San the same style to keep the city's personality.
Gimignano and New York-Manhattan The most representative images are the
silhouettes demonstrates the veracity of G.M. following: „Expulsion of the inhabitants from
Cantacuzino’s assertion. The message of both Carcassone in 1209” image from Grandes
profiles is aggressive, revealing the desire for Chroniques de France 1415; „La Délivrance
domination in a competition of economic des emmurés de Carcassonne” painting of Jean-
power. Paul Laurens 1879; old maps of Carcassone.
In order to deepen the argumentation about the The elaboration of the profile was based on
sketching and deciphering of built landscape several selected, compared and overlaid photos
profiles, the analyses were completed by two (Figures 3 a and b).
original case studies, with the extraction and
verification of the characteristic features of
some historic French cities.
The settlements selected on the basis of the
documentation were Carcassone and Castelnau-
le-Lez. In order that analyzes can be compared
as clearly and without confusion, the most
important criterion of the selection was that the
two cities must be different as human settle- Figure 3a. Drawing the Carcassone profile by
ments personality, in terms of environmental, overlapping the picture. Graphics Cojocariu M.
volumetric and urban tissue elements. (source: https://www.blog.thelittleprince.com/a-little-
An examination of characteristic images for prince-shop-in-the-medieval-city-of-carcassone-franc)
these cities was made and as a result, a few
vintage paintings, photographs and aerial
images were selected. The old images were
checked and compared to the actual photos and
on their basis the profiles of the two cities were
sketched. It was also necessary a document-
tation regarding the elements of the environ-
ment and on the evolution of the historical
stages.
The first case study is the historic city of Figure 3b. Final profile of Carcassone
Carcassone, which has some resemblance to (source: Graphics Cojocariu M.)
San Gimignano because it is fortified and has a
massive volume dominated by many towers. The energetic line of the silhouette highlights
It is a city located in southern France on the features characteristic of a defensive urban
hills of Languedoc. In contrast to the San organization. This strategic feature it is
Gimignano evolution, Carcassone has a rather certified by historical documents and is
agitated history with tragic episodes. confirmed by the qualities of landscape
The medieval citadel was built on the basis of a elements: the city is located on top of the hill,
preconceived urban plan being at present with a significant difference in level compared
protected by UNESCO. to the rest of the unfortified settlement, located
It was erected in the thirteenth century in order to the left and right of the fortress (Figure 3b).
to be an intangible fortress to defend the ever- The three important lines of the profile are: 1-
changing regional medieval frontiers, attacked the line from which the perception was made;
by infighting regional. 2-the citadel profile; 3-the background of the
The analysis of the historical pictures of the hills. The image of the profile on the
citadel highlights the fact that the ascending background of the surrounding hills makes a
characteristic features were similarly valuable asset of the medieval settlement,
578
namely its adaptation to the natural landscape with hilly relief, the river and the
environment. The line no. 2 stands out through vegetation (Figures 4b, 4c).
the multitude of defense towers, visible in the The final profile reveals the harmonious
density of the zigzags lines that evokes anxiety integration of all construction into the
of the defensive status. landscape, suggesting both the beauty of the
The second chosen case study is the small peaceful life and the attachment to nature.
settlement of Castelnau-le-Lez, located also in These are the most important features that have
southern France, on the river Lez, in the contributed to the development of the locality,
Occitan region. keeping a tradition of respect for history and
Research on the evolution of the site shows the natural environment.
that, dating from antiquity, the settlement was a
village with a quiet history and a continuous
economic development.
In terms of the natural environment, the
landscape is hilly with rich vegetation.
The overall image of the old area is picturesque
both as buildings and as a natural landscape.
From the historical images were selected, as the
most representative, two paintings done by the
impressionist artist Frédéric Bazille in 1864
and 1868 (Figure 4a). Both paintings have on
their background a gentle image of the quiet Figure 4b. Drawing the Castelnau-le-Lez profile by
settlement. Checking and comparing paintings overlapping the picture.
with current photos revealed that the old 19th (source: Graphics Cojocariu M.)
century core still retained its appearance.
Figure 4c. Removal of background paint

for profile decoding.
(source: Graphics Cojocariu M.)
These studies and researches revealed that by

analyzing only the old images and the present
photos as a whole we cannot decipher all subtle
Figure 4a. The final selected painting of Frederic features.
Bazille - View of the Village of Castelnau-le-Lez, 1868 Consequently, it was necessary to draw some
(source:https://www.wikiart.org/en/frederic-bazille/view-
of-the-village-of-castelnau-le-lez-1868) original profiles based on scientific
documentation.
The profile sketch was created by overlapping These analyze, in their entirety, highlighted the
the 1868 painting (Figures 4b and 4c). features of the personality of the studied
The three major lines for the basic features are: settlements, impregnated by the history of the
1-the river line; 2-the built profile; 3-the natural places and their natural landscape.
579
The sketching and deciphering of built Dascălu D.M., Cojocariu M., 2016. Landscape Design.
landscape profiles must be based on Ion Ionescu de la Brad Publishing House, Iaşi, 22-23.
Dascălu D.M., Dascălu V., 2017. The rescue of Iaşi city
comparative studies and analyses of the natural landscape-Homage to G.M. Cantacuzino. Scientific
environment, of the built and of socio- papers, Year LX, Vol. 60, No. 2, Horticulture Series,
economic-cultural environment. Ion Ionescu de la Brad Publishing House, Iaşi.
Elaborating silhouettes is especially useful if it Gassner G., 2009. Elevations, icons and lines: The city
is created by following the succession of abstracted through its skylines. In: Davis, Juliet, Dean,
Corinna, Gassner, Gunter, Hall, Suzanne and Keddie,
various historical periods. Jamie eds. Researching the Spatial and Social Life of
The decoding, analysis and comparative the City, Vol. 1. cities LAB, London School of
evaluation of the meanings transmitted by Economics and Political Science, 68-86
them, generates valuable information about the Gheorghiu T.O., 2002. Housing and Non-Housing.
essence, identity, personality and specificity of Paidea Publishing House, Bucharest, 45.
Gheorghiu T.O., 2009. Human settlements - Theoretical
human settlements. Highlights and History, vol. I, ArtPres Publishing
The research in this paper provides an original House, Timişoara, 36.
and particularly useful tool for studies of urban Reid W.G., 2007. From concept to form in Landscape
and landscape evolution of human settlements. Design, J. Wiley & Sons Publishing House, Canada.
Vasilescu S., 1993. Symmetry-Books and Criticism-
Dictionary (collection of texts). U.A.I.M. Publishing
House, Bucharest, 76-86.
***https://www.wikiart.org/en/frederic-bazille/view-of-
the-village-of-castelnau-le-lez-1868.
580
STUDIES REGARDING THE BEHAVIOUR OF ORNAMENTAL

SPECIES LAGURUS OVATUS IN CROPPING CONDITIONS
FROM N-E AREA OF ROMANIA
Elena Liliana CHELARIU1, Lucia DRAGHIA1, Maria BRÎNZĂ1, Mirela COJOCARIU1,

Bogdan-Vlad AVARVAREI2
1
Faculty of Horticulture, 3 Mihail Sadoveanu Alley, Iaşi, Romania
2
Faculty of Animal Science, 3 Mihail Sadoveanu Alley, Iaşi, Romania
Corresponding author eamail: [email protected]
Abstract
The current paper aimed to analyse some aspects about behaviour and multiplication of Lagurus ovatus species in
cropping conditions from Iaşi County, Romania. Research material was represented by Lagurus ovatus. Were
established five experimental variants represented by different substrate types utilised for sowing: V1 garden soil, V2
garden soil + leaves soil (1:1), V3 garden soil + leaves soil + peat (1:1:1), V4 leaves soil + peat (1:1) and V5 jiffy pots.
Observations and determinations were carried out in the didactic field of Floriculture Discipline from UASVM Iaşi,
Romania, during 2015-2016 and had in view aspects regarding seedlings obtaining and also plants behaviour. At the
end of the current research we observed that for seedlings production the best results were obtained at variant V3,
followed by V2 and V4; and in cropping conditions at variants V3, V2 and V4 the results were quite close as values. In
conclusion we can affirm that Lagurus ovatus is an ornamental grass which could be easily multiplied through seeds
and for establishing crops the utilisation of seedlings is recommended.
Key words: capitalization, Lagurus ovatus, ornamental (grass).
INTRODUCTION appearance and maturation took place in stages

from summer till autumn (Chelariu, 2013).
At world level plant species, generally named This species prefers fertile soils, permeable,
ornamental grasses, are utilised in floral art and with a pH between 6.6 and 7.5. Have a well
also in vegetal compositions in landscape development on sunny exposure fields (Brickell
designs. and Cathey, 2004; Chelariu, 2013; Colborn,
In Romania, ornamental grasses are less known 2006; Ondoño et al., 2016). Multiplication of
and utilised in landscape designs and in floral this species is realised on a generative way.
art. In the last years, interest for those species Seeds germination could be influenced by
increased, but due to lack of information different parameters of some ecological factors
regarding cropping technologies and utilisation (light, temperature, salinity etc.) (Carreño et al.,
modalities in pedo-climatic conditions of 2004).
Romania, their utilisation is not always In the current paper are presented the aspects
adequate (Chelariu, 2013). regarding seedlings obtaining and also the
Lagurus ovatus L. belongs to Poaceae behaviour in cropping conditions of Lagurus
botanical family and is cultivated in rustic areas ovatus species in pedo-climatic conditions from
8-10 (11) (Mills-Hicks, 2007). It is a species Iaşi, Romania.
with origins in South Europe, Mediterranean
area. It’s an annual species with a bush aspect, MATERIALS AND METHODS
height of 30-50 cm and leaves of a light green
colour (Brickell and Cathey, 2004; Şelaru, Research took place during 2015-2016 in the
2007). Flowers are grouped in small compact didactic collection (greenhouse and field) of
spikes, with a green white-yellow colour. Fruits Floriculture Discipline from UASVM Iaşi,
581
Romania. Biological material was represented by Holland. Experiments were organised in five
Lagurus ovatus species and the seeds used for variants, each variant being represented by a
establishing the experiments were achieved from substrate type utilised for sowing (Table 1).
Table 1. Experimental design
Studied species Biological material Seeds number Variant Substrate for sowing
Lagurus ovatus 100 V1 garden soil

Seeds
100 V2 garden soil + leaves soil (1:1)
garden soil + leaves soil + peat

100 V3
(1:1:1)
100 V4 leaves soil + peat (1:1)
100 V5 jiffy-pots
The experimental variants were established variant V5 (jiffy-pots) (Figure 1).

having in view the ecological demands of the
studied species and data from literature (Buta %
and Cantor, 2009; Cantor, 2009; Draghia and 100 95 100 95
90 90
Chelariu, 2011; Chelariu, 2013). 80 75
70
60
In each year sowing was realised on 10th of 50
40
February and the establishment of field crops 30
20
was done on 10th of May. 10
0
Research aimed the obtaining of seedlings V1 V2 V3 V4 V5
Va ri a nts
(seed germination percent, necessary period
from sowing till ending of plants’ emergence), Figure 1. Lagurus ovatus seeds germination (%)
as well as the behaviour in cropping in pedo-
climatic conditions from Iaşi, for species. From statistic point of view at variants V2, V3 and
In field, were studied separately plants obtained V4 were observed very significant positive
in different types of substrate, aiming aspects differences comparing with the control variant
regarding the growing rhythm, phenology and (V1). At variant V5 the negative difference value
morphology of plants. comparing with the control was very significant
The obtained results were statistically proces- (Table 2).
sed and were presented as a mean of two years Table 2. Results regarding seed germination
of study.
Number % face
RESULTS AND DISCUSSIONS Variant of seeds
(pieces)
to
control
Difference Signification
V1 90.0 100.00 0.0 control

Seeds‘ germination at species Lagurus ovatus V2 95.0 105.56 5.0 ***
could be influenced by light, temperature, soil V3 100.0 111.11 10.0 ***
salinity (Carreño et al., 2004). V4 95.0 105.56 5.0 ***
V5 75.0 83.33 -15.0 000
The research highlighted that the percent of LSD 5% = 1.4 pieces
germinated seeds at Lagurus ovatus varied LSD 1% = 2.0 pieces
LSD 0.1%= 3.1 pieces
between 75% and 100% (Figure 1). The best
results were recorded at variant V3 (1 part Regarding the necessary time period till plants’
garden soil + 1 part leaves soil + 1 part peat), emergence, were observed differences between
followed by variants V2 (1 part garden soil + 1 experimental variants. So, at variants V1, V2,
part leaves soil) and V4 (1 part leaves soil + 1 V3 and V4 the emergence started after 3 days
part peat), and the lowest values were obtained at from sowing and at variant V5 after 5 days
582
(Figure 2). Necessary time for a complete V2, 12 days at variants V1 and V4, for variant
emergence of plants, calculated from sowing, V5 were necessary 17 days from sowing to end
was of 10 days at variant V3, 11 days at variant of emergence (Figure 2).
V5 5 12
V4 3 9
V3 3 7
V2 3 8
V1 3 9
0 number of days 5 10 15 20
time interval from sowing to starting of germination

time interval till germination ending
Figure 2. Duration of germination (number of days from sowing)
The utilised seedlings for establishing the field Table 4. Phenology of Lagurus ovatus specie in cropping
crops had different features function by conditions from Iaşi County
experimental variant (Table 3). Flowering Fructify
Ending of
Growing vegetation
Variant (decade/ (decade/
Table 3. Morphological features of seedlings planted in rhythm
month) month)
period
experimental field (decade/month)
V1 normal II/06 II/08 I/10
Mean
Mean length Mean Mean V2 normal II/06 II/08 I/10
number of
Variant of seedlings height of number V3 normal II/06 II/08 I/10
roots per
(cm) plant (cm) leaves/plant
plant V4 normal II/06 II/08 I/10
V1 6.3 14.2 11.2 14.4
V5 normal I/07 III/08 I/10
V2 6.8 14.4 11.6 14.6
V3 7.2 15.1 12.5 14.8
V4 6.7 14.3 12.0 14.7 Table 5. Morphological characterization of Lagurus
V5 5.4 9.6 8.1 13.0 ovatus plants Mean number
inflorescence
inflorescence
Seedlings obtained in greenhouse, on different

Mean height
Mean length
Mean length
Mean width
Diameter of
of leaves
of leaves
per plant
-pieces-
of plant
Variant
types of substrates were planted in field on 10th

bush
-cm-
-cm-
-cm-
-cm-
-cm-
of
of May. In according with literature (Brickell

and Cathey, 2004; Colborn, 2006; Ondoño et V1 38.8 23.3 14.1 1.3 5.3 24.3
al., 2016) plants of Lagurus ovatus reach a V2 40.1 23.5 14.2 1.3 5.4 24.6
height between 30 and 50 cm; flowering in V3 40.9 24.2 14.5 1.4 5.5 25.1
summer. During research period, in pedo- V4 40.3 23.9 14.4 1.4 5.5 25.0
climatic conditions from Iaşi, Romania, plants V5 34.9 21.9 14.0 1.3 5.3 23.9
had a normal rhythm of growing. Flowering
took place in the second decade of June (V1, Regarding mean height of plants were observed
V2, V3, V4) or in the third decade of June (V5), very significant positive differences comparing
and fructification in the second decade (V1, V2, with the control (V1) at variant V3 and distinct
V3, V4) or in the third decade of August (V5). significant positive differences at variants V2 and
Vegetation period ended in the first decade of V4. At variant V5 the negative difference
October (Table 4). comparing with the control variant is very
In Table 5 is presented the morphological significant (Table 6).
characterization of plants, realised based on Regarding flowering capacity at variants V3
field observations made on 1st of October. and V2 differences comparing with the control
583
(V1) were positive distinct significant while recommended as substrate for sowing the
variant V5 recorded a significant negative mixture between garden soil + leaves soil +
difference (Table 7). peat (1:1:1).
In crop conditions, at plants obtained at
Table 6. Results regarding plants’ growing
variants V3, V2, V1, and V4, morphological
Mean % face characteristics were quite similar, differences
Variant height of to Difference Signification
plant (cm) control being observed at plants from variant V5.
V1 38.8 100.00 0.0 control Lagurus ovatus is an ornamental grass species
V2 40.1 103.35 1.3 ** which in cropping conditions from North-East
V3 40.9 105.41 2.1 *** area of Romania acts as an annual plant, having a
V4 40.3 103.87 1.5 ** normal growing rhythm and decorates through
V5 34.9 89.95 -3.9 000 habitus and inflorescences.
LSD 5% = 0.8 cm
LSD 1% = 1.1 cm
LSD 0.1% = 1.7 cm REFERENCES
Table 7. Mean number of inflorescences/plant Brickell Ch., Cathey M., 2004. The American Horticultural
Society A-Z Encyclopaedia of Garden Plants. The
American Horticultural Society. Published by DK
Signification
Publishing Inc. ISBN 0 7566 0616 0, 589.

Difference
Number of
% face to
Variant inflorescences
control Cantor M., Buta E., 2009. Floricultură - îndrumător de
(pieces) lucrări practice. TODESCO, Cluj-Napoca. ISBN 978
973 7695 84 0, 108-109.
V1 24.3 100.00 0.0 control Cantor M., 2009. Floricultură generală. TODESCO,
V2 24.6 101.23 0.3 - Cluj-Napoca.
V3 25.1 103.29 0.8 ** Carreño S., Contesa E., Franco J.A., Martínes-Sánchez
V4 25.0 102.88 0.7 ** J.J., 2004. Effect of temperature and salinity on the
V5 23.9 98.35 -0.4 0 germination of Lagurus ovatus L. Hort. Science
LSD 5% = 0.4 pieces vol.39(4),http://hortsci.ashspublications.org/content
LSD 1% = 0.5 pieces
LSD 0.1% = 0.8 pieces /39/4/836.2.short.
Chelariu E.L., 2013. Evaluation and completing the
ornamental grasses assortment from the N-E area of
In landscape design, Lagurus ovatus assures Romania. Research project in the project number
decoration due to its bush habitus, from spring till POSDRU/I.89/1.5/S62371 ,,Postdoctoral School in
autumn, and by inflorescences in period June- Agriculture and Veterinary Medicine area”, UASVM
September (Chelariu, 2013). Due to fine texture Iaşi, Romania.
and plants’ elegance these species could be Colborn N., 2006. Plant solutions. Harper Collins
Publishers, United Kingdom. ISBN -13: 978 0 00
utilised in vegetal compositions from different 7193127, p.21, 215.
types of landscape designs such as: decorative Draghia L., Chelariu E.L., 2011. Floricultură. Editura
pots, rounds, flats, stone designs etc. (Brickell „Ion Ionescu de la Brad”, Iaşi. ISBN 978 973 147 074
and Cathey, 2004; Chelariu, 2013; Colborn, 0, p. 121, 168, 226.
2006; Şelaru, 2007; Tomaškin et al., 2015), Mills-Hicks J., 2007. Botanica’s Pocket Gardening
Encyclopaedia. Published by H.F. Ullmann, Australia.
green roofs (Ondoño et al., 2016). Also, floral ISBN 978 3 8331 2939 1, 669.
rods could be utilised in floral art as cut flowers Ondoño S., Martínez-Sanchez J.J., Moreno J.L., 2016.
both in fresh state as well as in dry state The composition and depth of green roof substrates
(Chelariu, 2013; Tomaškin et al., 2015). affect the growth of Silene vulgaris and Lagurus
ovatus species and the C and N sequestration under
two irrigation conditions. Journal of Environmental
CONCLUSIONS Management, vol. 166, 330-340.
https://www.sciencedirect.com/science/article/pii/S03
In conclusion we can affirm that Lagurus 01479715302498.
ovatus is an ornamental grass which could be Tomaškin J., Tomaškinova Judita, Kizeková Miriam,
easy multiplied through seeds and for 2015. Ornamental grasses as part of public green, their
ecosystem sercices and use in vegetative arrangements
establishing of crops is recommended the in urban environment. Thaiszia-J. Bot. Kosice, 25 (1):
utilisation of seedlings. 1-13, http://www.bz.upjs.sk/thaiszia.
To obtain a 100% percent of germination and a Şelaru E., 2007. Cultura florilor de grădină. Editura
quality seedling at species Lagurus ovatus is CERES, Bucureşti. ISBN 978-973-40-0774-5, 510
584
THE RESPONSE OF BULGARIAN SPRAY-CARNATION

(D. CARYOPHYLLUS F. SPRAY, HORT.) CV. ʻRUSALKAʼ TO DROUGHT -
IN VITRO INDUCED BY DIFFERENT PEG CONCENTRATIONS
Violeta IVANOVA1, Nadejda ZAPRIANOVA2, Bistra ATANASOVA2, Valentin PANCHEV1

1
2
Institute of Ornamental Plants, Negovan 1222, Bulgaria
Corresponding email: [email protected]
Abstract
In our study, to simulate water deficit induced by osmotic stress, different concentrations of polyethylene glycol (PEG-
6000) were used: 10%, 20%, 30% and 40% at different durations of treatment (1, 3 and 6 days) in vitro conditions. The
model plant was Bulgarian spray-carnation (D. caryophyllus f. spray, Hort.) flowers, cv. ʻRusalkaʼ. The response to
drought stress was studied based on the following end-points: plant growth reactions, relative water content (RWC %),
and electrolyte leakage (conductivity). The water deficit varied from 16% (control) to 75% (40% PEG-6 days). The
growth of the explants proportionally decreased with the increase of polyethylene glycol concentration from 10% to
40% and the fresh weight was below 50% vs. the control at 30% and 40% PEG. The relative water content of the plant
tissues decreased depending on PEG quantity, the lowest values - 25.16 ± 2.06% being reported at 40% PEG
concentration on the 6th day. The highest values of electrolyte leakage up to 1712 µS/g fresh weight were reported on
the 6th day at 40% PEG concentration.
Key words: spray-carnation, drought, polyethylene glycol (PEG), water deficit, growth.
INTRODUCTION response of the plants to stress on cellular

level-in vitro and in vivo (Song et al., 2013)
The ongoing worldwide climate changes The purpose of the investigation was to
enforced multiple studies of how the plants establish the physiological and adaptive
react towards them. One of the main response of the Bulgarian spray-carnation
components of the drought is the water deficit, cultivar to drought.
an abiotic factor that causes multiple
morphological and physiological changes in the MATERIALS AND METHODS
plants that reduce their quality and economic
value. For the purpose of the experiment, the plant
The research on the physiological mechanisms material of the D. caryophyllus f. spray, Hort.)
of plant resistance in laboratory conditions cv. ʻRusalkaʼ was reproduced in vitro on MS
gives an opportunity to monitor the specific nutritive medium with added sacharose - 30 g/l
response of the plants to one of the impact and agar 6 g/l at pH = 5.7-5.8 prior to
factors (Yordanov et al., 2000; Alexieva et al., autoclaving (Murashige and Skoog, 1962).
2003). Then it was grown in a phytostatic room at a
The drought simulation was done by means of temperature of 22C, photoperiod of 16: 8 (day:
an osmotic agent with high molecular weight night) hours and light intensity 30 mol m-2 s-1.
(>3000) such as polyethylene glycol (PEG) For the induction of the experiment, an MS
(Murillo Amador et al., 2002). The use of PEG medium was used containing salts and
in liquid media allowed achieving precisely and vitamins, saccharose - 30 g/l and polyethylene
recreating the necessary osmotic potential of glycol (PEG 6000) in the following
the environment (Song et al., 2013). concentrations - 10%, 20%, 30% and 40% with
The response of the in vitro cultures to induced pH = 5.7-5.8 prior to autoclaving.
stress enabled the selection of water deficit The explants were placed in test-tubes on
tolerant plants at an early stage. This was made control (0) and stress inducing liquid medium
possible by the existing correlation in the (PEG - 10%, 20%, 30% and 40%) on filter
585
paper bridges, 10 for each concentration and The growth decreased proportionally to the
the control in 3 replications. The duration of increase of PEG concentration in the nutritive
stress impact was short (one day), medium medium, it was less than 50% vs. the control in
(three days) and long-term (six days). the high PEG concentrations (30% and 40%),
The explants used were 2-3 cm long and had a 58.94 ± 4.4 and 59.96 ± 2.5, respectively, with
weight of 100-300 mg, measured in sterile significance rate P<0.0001. Following short-
conditions prior to the initiation of the stress term (one-day) osmotic stress, the explants
inducing medium. showed little growth demonstrated by its gain
In order to establish the effect of water deficit in fresh weight (Figure 1).
on plant tissues, we measured the explants' The results were statistically significantly
growth and rooting, the rate of cell membranes different at P<0.05 (*), P<0.01 (**), P<0.0001
damage and the relative water content. (***), respectively, as compared to the control.
In in vitro research, growth was expressed as The reported growth values were 33.78 ± 6% at
the percentage of micro explants’ weight 30% PEG and 30.32 ± 2.7% at 40% PEG in the
increase after being cultivated for a certain extended treatment periods (3 and 6 days)
period of time (1, 3 and 6 days) on 10%, 20%, (Figure 1). The control plants were fresh, green
30% and 40% PEG, compared to the initial and in normal turgor condition and showed
weight. initial rooting on the 6th day. The plants placed
The rate of the membrane damage was defined on 10% and 20% PEG concentration showed
by the electrolyte leakage from the leaves with slight wilting, compared to the higher wilting
accounting for conductivity only after stress rate on 30% and 40 % and no explants necrosis.
and was expressed as µS/g fresh weight. At higher PEG concentrations and longer
The relative water content (RWC) was exposure times (3-6 days), a progressive
measured simultaneously with electrolyte decrease in the growth was observed: down to
leakage and calculated by the following 50% below the control at 30% and 40% PEG.
formula: The control plants were fresh, green and with
RWC % = (fresh weight – dry weight)/(turgor normal turgor on day 6 while the plants
weight – dry weight) × 100 - according to exposed to 10% and 20% PEG showed
Turner’s method (Turner, 1981). withering signs which were even more notable
The water deficit (WD) was expressed by the in the 30% and 40% PEG plants.
following formula: The suppression of growth was related to the
WD % = 1 – RWC. reduced capacity of the plant to uptake water
Following the stress period, the explants were (Shabani et al., 2013). The effect of the osmotic
transferred to an MS medium without a stress stress on cellular level is expressed by slowing
agent (PEG) in order to establish their capacity down cell division, they lose their turgor and
for recovery and report the rooting percentage. this leads to weight loss (Levitt, 1980; Heyser
The data on the figures below were expressed and Nabor, 1981). Due to the low turgor
as an average value ± SE of two independent pressure cell expansion and cell growth
experiments, carried out in 10 replications per suppresses under water stress (Jaleel et al.,
variant. They were analyzed for significance by 2009). In chrysanthemum, the values of the
means of the t-test of the GraphPad Prizm fresh weight decreased up to 50% vs. the
software. The results were statistically control. The plants were wilting in low PEG
significantly different at P<0.05 (*), P<0.01 concentrations (10% and 20%), while 20% of
(**), P<0.0001 (***), respectively, as the trial explants became necrotic in addition to
compared to the control. wilting in the high concentrations (30% and
40%) (Zapryanova and Nencheva, 2013).
RESULTS AND DISCUSSIONS In Bulgarian spray carnation (D. caryophyllus
f. spray, Hort.) flowers, cv IRA a progressive
Following the exertion of a short-term osmotic decrease in the growth was observed: down to
stress (1 day), the explants, Grown on the 50% below the control at 30% and 40% PEG
control medium, showed a slight growth and (Zapryanova et al., 2015).
reached values up to 102.8 ± 5.8 (Figure 1).
586
induced stress led to a gradual decrease in the
explants’ RWC. The lowest RWC values
(28.52 ± 5.2) were obtained in the 40% PEG
variant on day 6. Prolonged drought resulted in
low RWC of cells and in low tissue turgor and
irreversible withering, especially at higher
concentrations (Zapryanova et al., 2015).
Fig. 1. Growth (gain in fresh weight) of spray-carnation

explants cv. ʻRusalkaʼ in vitro at different PEG
concentrations and stress duration (1, 3 and 6 days)
The index of the Relative water content (RWC)

is often used to define the water status. It
decreases with the increase of the PEG
concentrations in plant tissues. The control
plants maintained a high percent of water
content in their tissues - about 80% (Figure 2). Fig. 2. Relative water content (RWC) of spray-carnation
The results of day 1 showed that the exerted explants cv. ʻRusalkaʼ in vitro at different PEG
PEG stress lead to the gradual decrease of plant concentrations and stress duration (1, 3 and 6 days)
water content, the lowest values of 44.62 ± 2.3,
being reported at 40% PEG concentration. The The damage of cell membrane organization and
difference between the separate variants and content is one of the first responses of the plant
the control were statistically significant at organism to the impact of the stress agent.
P<0.0001. That tendency was maintained on Electrolyte leakage in control plants was
both the 3rd (29.25 ± 2.3) and 6th days (25.16 ± observed in low rates on the 1st day after trial
2.06) lowest results for PEG. The results for all initiation and was maintained throughout the
PEG concentrations vs. the control were trial duration (µS/g fresh weight): 396.78 ±
statistically significant at P<0.05 and P<0.0001 54.8 - 1st day, 226.53 ± 27.9 - 3rd day and
(Figure 2). 292.478 ± 37.58 - 6th day µS/g fresh weight that
A similar response was observed in callus corresponded to the normally expected
culture of Carthamus tinctorius L. - the lowest response of the studied material.
growth values and RWC percentage were The simulated experimental drought showed a
reported for 40% PEG concentration (Kakaei et sharp increase of electrolyte leakage during
al., 2013). short-term stress (1 day) in all the used PEG
The long-term drought stress in concentrations (Figure 3). The highest values –
chrysanthemum strongly reduced the water 1712 ± 363 µS/g fresh weights were reported at
potential of the cell, lead to the decrease of 40% PEG concentration - 6th day (Figure 3).
tissue turgor and final wilting of the plants, The values of the electrolyte leakage at
especially in the higher PEG concentration. different PEG concentrations during the 3 and 6
Whereas the values of the control plants were days stress were constantly higher than the
constant at about 70%, they reached 30% on control plants but lower in comparison to the
the 6th day at 40% PEG concentration results of the short-term stress. The results had
(Zapryanova and Nencheva, 2013). a good statistical significance between the
In experiments with Bulgarian spray carnation separate variants and the control at P<0.05 and
(D. caryophyllus f. spray, Hort.) flowers, cv. P<0.0001 (Figure 3).
ʻIRAʼ, the control explants (without PEG) Electrolyte leakage in the Bulgarian spray
maintained high RWC in their tissues: about carnation (D. caryophyllus f. spray, Hort.)
80%, regardless of the cultivation time (1, 3 or flowers, cv IRA - control plants was very low
6 days). The results clearly indicate that PEG- at day 1 and remained under 200 µS g-1 during
the whole experiment, which is in good
587
correspondence with the expected normal The spray-carnation explants were placed on
values for the chosen plant material. The MS medium after the different stress periods
highest values 2633 ± 521 µS/g fresh weight for recovery. The results showed that the
were reported at 40% PEG concentration recovery is 100% after the short-term stress (1
(Zapryanova et al., 2015). day). Rooting was reported for all the
concentrations, regardless of PEG quantity:
100% for 10% and 20% PEG; 75% for 30%
PEG and 60% rooting - for 40% PEG.
The plants recovered 100% after the medium -
term stress (3 days) at PEG concentrations -
10%, 20% and 30%, while only 90% recovered
at 40% PEG concentration. The best
manifestation of the rooting - 90% - was at
10% PEG, where it was expressed to a smaller
degree at the higher PEG concentrations of
30% - 50% rooting and 40% - 30% rooting.
The long-term 6 days stress of PEG showed
100% recovery of the plants at 10%, 20% and
30% PEG concentrations but the percentage of
rooted plants went down to 50. The recovery at
Fig. 3. Electrolyte leakage from spray-carnation higher PEG concentrations of 40% was 30%
explants cv. ʻRusalkaʼ in vitro at different PEG and rooting only 10%.
concentrations and stress duration (1, 3 and 6 days) The explants of spray-carnation cv. ʻIRAʼ
showed a good adaptive response that was
The highest values of water deficit (WD %) confirmed by the high recovery percentage -
were reported during the long-term stress (6 60% and 40%, reported for the high PEG
days period), namely 69% and 76% at 30% and concentrations - 30% and 40%, respectively
40% PEG (Figure 4). These results indicate that (Zapryanova et al., 2015).
the drought stress induced in our experiments In conditions of moderate PEG-induced
was of a moderate degree but developed drought stress- in vitro, the explants of
rapidly. According to the concept of Cornic G., Bulgarian chrysanthemum cv. ʻZhoroʼ showed
Fresneau C. (2002), dehydration that causes up an adaptive response, rather than stress-induced
to 30% water deficit in plants, is assumed as damage, as inferred from the relatively high
mild or moderate stress. recovery rate (80% and 60%) following 30%
A similar response was observed in and 40% PEG treatment (Zapryanova and
experiments with Bulgarian spray carnation Nencheva, 2013).
(D. caryophyllus f. spray, Hort.) flowers, cv.
ʻIRAʼ. Dehydration of plant tissues to 69% for CONCLUSIONS
the PEG 30% variant and to 71% for the PEG
40% variant on day 6th was observed, whereas The growth of the explants proportionally
the values on day 1 were 46.2% and 63.62%, decreased with the increase of polyethylene
respectively (Zapryanova et al., 2015). glycol concentration from 10% to 40% and the
fresh weight was below 50% vs. the control at
30% and 40% PEG.
The drought, simulated by means of different
polyethylene glycol concentrations, caused
changes in the cell membranes of spray-
carnation cv. ʻRusalkaʼ. The highest values of
electrolyte leakage up to 1712 ± 363 µS/g fresh
weight were reported on the 6th day at 40%
Figure 4. Water deficit (%) in the plant tissues of spray- PEG concentration.
carnation explants cv. ʻRusalkaʼ in vitro at different PEG
concentrations and stress duration (1, 3 and 6 days)
588
The relative water content of the plant tissues Levitt J., 1980. Responses of plants to environmental
decreased depending on PEG quantity, the stresses. Volume II. Water, radiation, salt, and other
stresses (No. Ed. 2). Academic Press.
lowest values – 25.16 ± 2.06% being reported Murashige T., Skoog F., 1962. A revised medium for
at 40% PEG concentration on the 6th day. rapid growth and bioassays with tobacco tissue
The water deficit varied within 18% - 76% cultures Physiol. Plant.15, 473-497.
depending on PEG concentration and durations Murillo Amador B., Lopez-Aguilar, Kaya R., Larrinaga-
of treatment. Mayoral J., Flores-Hernandez A., 2002. Comparative
effects of NaCl and PEG on germination, emergence
The explants of spray-carnation cv. Rusalka and seedling growth of cowpea. J. Agron. Crop
showed a good adaptive response that was Sci.188, 235-247.
confirmed by the high recovery percentage - Shabani A, Sepaskhah A.R., Kamgar-Haghighi A.A.,
100% and 30 %, reported for the high PEG 2013. Growth and physiologic response of rapeseed
concentrations - 30% and 40%, respectively on (Brassica napus L.) to deficit irrigation, water salinity
and planting method. Inter. J. of Plant Production 7,
the 6th day. 569- 596.
Song H., Seo Y., Jeong M., Kim H., Im H., Cho H., Choi
REFERENCES M., 2013. In vitro evaluation system using osmotic
agents of drought tolerance ecological restoration
Alexieva V., Ivanov S., Sergiev I., Karanov E., 2003. plants. Korean Institute Of Forest Recreation and
Interaction between stresses. Bulg. J. Plant Welfare, 4, 611-613.
Physiol.Special Issue.1, 18. Turner N.C., 1981.Techniques and experimental
Cornic G., Fresneau C., 2002. Photosynthetic carbon approaches for the measurement of plant water stress.
reduction and oxidation cycles are the main electron Plant Soil. 58, 339-366.
sinks for photosystem II activity during a mild Yordanov I., Velikova V., Tsonev T., 2000. Plant
drought. Ann. Bot. 89, 887-894. responses to drought, acclimation, and stress
Heyser J., Nabors M., 1981. Growth, water content, and tolerance Photosynthetica 38, 171-186.
solute accumulation of two tobacco cell lines cultured Zapryanova N., Atanassova B., Ivanova V., 2016. Effect
on sodium chloride, dextran, and polyethylene glycol. of water deficit induced by osmotic stress on
Plant physiology, 68(6), 1454-1459. Bulgarian spray-carnation (D. caryophyllus F. Spray
Jaleel C., Manivannan P., Wahid A., Farooq M., Al- Hort.) cv. Ira in vitro. Dekoratyviuju ir sodo augalu
Juburi H., Somasundaram R., Panneerselvam R., sortimento, technologiju ir aplinkos optimizavimas
2009. Drought stress in plants: a review on Optimization of Ornamental and Garden Plant
morphological characteristics and pigments Assortment, Technologies and Environment, 7(12),
composition. Int J Agric Biol, 11(1), 100-105. 146-151.
Kakaei M., Mansouri M., Abdollahi M.R., Moradi F., Zapryanova N., Nencheva D., 2013. Effect of water
2013. Effect of NaCl and PEG induced osmotic stress deficit induced by osmotic stress on bulgarian
on callus growth parameters of two Safflower chrysanthemum cv. Zhoro in vitro Subtropical and
(Carthamus tinctorius L.) cultivars. Intl J Agri Crop Ornamental Horticulture, 49, 253-260.
Sci. Vol. 6 (3), 127-132.
589
590
ASPECTS REGARDING THE ORNAMENTAL VALUE OF SOME ROSE

NURSEY SPONTANEOUS IN IAŞI DISTRICT CONDITIONS
Roberto BERNARDIS, Marius DASCĂLU
3 Mihail Sadoveanu Alley, Iaşi, Romania
Abstract
Wild species of the genus Rosa L. have importance for: creating green spaces, getting roses nobles and their use as
rootstock for cultivated varieties. The purpose of this paper is to highlight the possibilities of identifying potential
ornamental species in the genus Rosa sp. Observations were made at 6 species of wild rose “Rosa californica”, “Rosa
caudata”, “Rosa centifolia”, “Rosa damascena”, “Rosa multiflora” and “Rosa rugosa”. To achieve that goal were
made observations and measurements on the main morphological characters, growth of stems, number of buds and
abundance of flowers.
Key words: roses, stems, buds, flower.
INTRODUCTION was proposed to be realised phenological

observations on: stems, leaf, flowers and fruits.
One of the basic components of green spaces
which assure a esthetical aspect of localities and MATERIALS AND METHODS
contributes to the welfare and good mood of
people, which also assure a favourable working Observations on roses were carried out at
and living climate is represented by floral and “Tudor Neculai” - Iaşi nursery. Nursery is
roses landscapes designs (Wagner Șt., 2002). placed outside Iaşi City, into a field belonging
Rose was considered from ancient times “Queen to Miroslava village, being destined for
of flowers”, due to its multiple qualities and production of planting dendrological and floral
particularly great wealth and beauty flowers, material for decoration and beautification of
scented and with various colours and shapes Iaşi City green spaces. Nursery have also a rich
(Zaharia D. et al., 2003; Wagner Șt., 2002). assortment of spontaneous rose species from
In the multitude of dendrological species which which we mention: Rosa californica, Rosa
contains ornamental shrubs, spontaneous canina, Rosa carolina, Rosa caudata, Rosa
species of Rosa L. have a real importance for centifolia, Rosa corymbifera, Rosa damascena,
design of green spaces, obtaining of noble Rosa davidii, Rosa davurica, Rosa foetida,
roses, and many of them are utilised as parent Rosa gallica, Rosa multiflora, Rosa rugosa,
stocks for cultivated sorts. (Mohan et al., 2010) Rosa virginiana (Bernardis R., 2011; Zaharia
The conditions from Romania are generally D. et al., 2003; Wagner Șt., 2002).
favourable for this specie. Observations were made on 6 spontaneous rose
Necessity for diversifying the assortments is a species:
priority, having in view the diversity of - Rosa californica (Figure 1);
biological material and the results obtained in - Rosa caudata (Figure 2);
the world. Literature shows, which in this - Rosa centifolia (Figure 3);
genus are known over 400 species with more - Rosa damascena (Figure 4);
than 20,000 sorts (Bernardis R., 2011; Mohan - Rosa multiflora (Figure 5);
et al., 2010; Zaharia D. et al., 2003; Wagner - Rosa rugosa (Figure 6).
Șt., 2002). The aim of the current paper is to highlight the
Being known the diversity importance of Rosa ornamental potential and identification possibi-
L. spontaneous species, in the research method
591
lities of Rosa spontaneous species in the genus
(Bernardis R., 2011; Milică C. et al., 2010).
Were realised observations and determinations
regarding: growing rhythm of stems; number of
buds; abundance of flowers.
Observations and determinations regarding
phenological particularities were realised for
six Rosa L. spontaneous species, in three
repetitions and in five different days, as
follows: (28.04, 7.05, 9.06, 11.07 and 10.08).
Observations were carried out during 2015-
2016 and were studied the growing rhythm of
stems, number of buds and abundance of Figure 4. Rosa damascena
flowers.
Figure 5. Rosa multiflora

Figure 1. Rosa californica
Figure 6. Rosa rugosa

Figure 2. Rosa caudata
From measurements made at the end of

observation period (10.08) at analysed species
regarding height of stems were recorded low
growing rates of 11.33 cm at Rosa californica,
14 cm at Rosa rugosa, and the greatest growing
rates were recorded at Rosa damascena with a
stem growing of 19.66 cm and 16.33 at Rosa
caudata (Table 1).
Figure 3. Rosa centifolia
592
Table 1. Average of stem height (cm) during Table 3. Average of flowers abundance during
28.04.2015 - 10.08.2015 28.04.2015 - 10.08.2015
Nr. Species 28.04 7.05 9.06 11.07 10.08 No Species 28.04 7.05 9.06 11.07 10.08
Rosa Rosa
1 5.33 32 11.66 9.03 11.33 1 - - 1.66 3.33 1.33
californica californica
Rosa Rosa
2 6 7.33 13.66 10.66 16.33 2 - - 2.33 - -
caudata caudata
Rosa Rosa
3 6 7.66 21.66 1.8 15.33 3 - - 3.66 - -
centifolia centifolia
Rosa Rosa
4 5 12.66 13.66 14 19.66 4 - - 2 2.33 8.66
damascena damascena
Rosa Rosa
5 12 12 15 9.66 15.66 5 - - 8 3 4
multiflora multiflora
Rosa Rosa
6 11.66 5.33 14.33 13.66 14 6 - - 1.33 2.33 3
rugosa rugosa
Analysing the number of buds, the lowest Table 4. Total average stems heigt, buds number,
values were recorded at specie Rosa californica flower abundance during 28.04.2015 - 10.08.2015
(4.66), and the greatest values were recorded at Height
species Rosa multiflora (8.66) and at Rosa Nr. Species of stems
Number Abundance of
of buds flowers
damascena (10.66) (Table 2). (cm)
Rosa
1 13.93 3.53 1.26
californica
Table 2. Average of buds number during
Rosa
28.04.2015 - 10.08.2015 2 10.79 0.86 0.46
caudata
No Species 28.04 7.05 9.06 11.07 10.08 Rosa

3 13.66 1.26 0.73
centifolia
Rosa Rosa
1 1 1 4.33 6.66 4.66 4 13 4.26 2.60
californica damascena
Rosa Rosa
2 - 3.33 1 - - 5 21.44 6 3
caudata multiflora
Rosa Rosa
3 1 2 3.33 - - 6 11.79 3.46 1.33
centifolia rugosa
Rosa
4 - 1.33 3.33 6 10.66
damascena
Rosa Table 5. Average stems height, buds number,
5 - 3.33 11.33 6.66 8.66 flower abundance during 9.06.2015 - 9.06.2016
multiflora
Rosa
6 1 1 3 5.33 7 Average at 9.06.2015 Average at 9.06.2016
rugosa
Buds number
Buds number
Stems height
Stems height
abundance
abundance
No. Species
Study realised regarding recorded abundance of
Flowers
Flowers
(cm)
(cm)
flowers at the end of observation period

appreciate that the lowest values were recorded
Rosa
by Rosa californica (1.33) and the highest 1
californica
11.66 4.33 1.66 13.33 6.33 2.33
values were recorded at Rosa damascena (8.66) Rosa
2 13.66 1 2.33 15 2 4
and Rosa multiflora (4) (Table 3). caudata
Also was realised a total mean regarding stems 3
Rosa
21.66 3.33 3.66 24 5.33 5.33
centifolia
height, buds number and flowers abundance in Rosa
4 13.66 3.33 2 15.66 5 3.33
period 28.04.2015 and 10.08.2015 (Table 4) damascena
and average for period 09.06.2015 - 09.06.2016 5
Rosa
15 11.33 8 16.66 13 10.33
multiflora
(Table 5), and based on them were designed 6 Rosa
graphs for year 2015 (Figures 7-12) and 6 6
rugosa
14.33 3 1.33 15.66 4.33 3
graphs for year 2016 (Figures 13-18).
593
”Rosa californica” ”Rosa multiflora”
1.66 8
4.33 Stems height Stems height
15
11.66 Buds number Buds number
11,33
Flower abundance Flower abundance
Figure 7. Average stems height, buds number, Figure 11. Average stems height, buds number,
flowers abundance at 09.06.2015 flowers abundance at 09.06.2015
for Rosa californica variety for Rosa multiflora variety
”Rosa caudata” ”Rosa rugosa”
2,33 3 1,33
1 Stems height Stems height
13,66 Buds number Buds number

14,33
for Rosa caudata variety for Rosa rugosa variety
”Rosa centifolia” ”Rosa californica”
3,66
2,33
3,33
Stems height Stems height
Buds number 6,33
21.66 13,33 Buds number
for Rosa centifolia variety for Rosa californica variety
”Rosa damascena” ”Rosa caudata”
2 4
2 Stems height
3,33 Stems height 15
Buds number
13,66 Buds number
Flower abundance
Flower abundance
Figure 14. Average stems height, buds number,

Figure 10. Average stems height, buds number, flowers abundance at 09.06.2016
flowers abundance at 09.06.2015 for Rosa caudata variety
for Rosa damascena variety
594
”Rosa centifolia”
CONCLUSIONS
Based on the effectuated determinations,

5,33 observations and analyse of the obtained results
Stems height
5,33 we could draw the following conclusions:
24 Buds number 1. Regarding the stems growing rhythm:
Flower abundance - spontaneous species of Rosa have a different
growing rhythm;
- the highest growing rate: Rosa multiflora with
an annual mean of 21.4 cm, and the lowest one
flowers abundance at 09.06.2016 at Rosa caudata 10.79 cm.
for Rosa centifolia variety 2. Regarding number of buds:
- buds number is different function of their
appearance period on stems and function of
”Rosa damascena”
specie;
3,33
- at Rosa californica, Rosa centifolia and Rosa
Stems height rugosa, buds appear in the second decade of
5
15,66 Buds number April and have a mean of 1 bud. At Rosa
Flower abundance
multiflora and Rosa caudata buds appear on
stems in the first decade of May with a mean of
3.33 buds.
3. Regarding abundance of flowers:
Figure 16. Graphs regarding average stems height,
buds number, flowers abundance at 09.06.2016 - flowers differs through abundance and
for Rosa damascena variety decoration period;
- Rosa centifolia, decoration period is in June
with a flowers abundance mean of 3.66, and
”Rosa multiflora”
Rosa damascena have a decoration period
between June-July-August with an average of
10,33 2.60.
16,66 Stems height 4. Regarding development of Rosa spontaneous
13
Buds number species between 9.06.2015 and 9.06.2016 we
Flower abundance could appreciate that:
- growing rhythm of stems is between 1.33 cm
(Rosa rugosa) to 2.34 (Rosa centifolia);
- buds number in these period increase in
flowers abundance at 09.06.2016 average from 1 (Rosa caudata) to 2 (Rosa
for Rosa multiflora variety centifolia and Rosa californica);
- flowers abundance at majority of studied Rosa
species increases with a mean value around
”Rosa rugosa”
1.67.
Having in view the tradition of rose cultivation,
3 their biological potential and high decorative
4,33 Stems height value, it is recommended utilisation of Rosa
15,66 Buds number genus spontaneous species for:
Flower abundance - parent stock for noble sorts;
- decorative purposes in the majority of
landscape designs.
flowers abundance at 09.06.2016 REFERENCES
for Rosa rugosa variety
Bernardis R., 2011. Ornamental arboriculture. Vol. I.,
”Ion Ionescu de la Brad” Publishing House, Iași, 119-
130.
595
Milică C., Bavaru A., Bavaru Elena, 2010. Roses Zaharia D., Dumitraș Adelina, 2003. Ornamental
species: tenderness, beauty, health: The Rosa genre arboriculture. Risoprint Publishing House, Cluj
monography. Pim Publishing House, Iași, 89-95. Napoca, 304-305.
Mohan Gh., Ardelean A., 2010. Decorative plants Wagner Şt., 2002. The Rose from mith to the third
encyclopedia. Vol. I., Trees and bushes. All millennium. Echard et Co. SNC Publishing House,
Publishing House, București, 293-300. Cluj-Napoca, 21-27.
596
STUDIES ON GROWTH AND DEVELOPMENT OF HEDERA HELIX L.

ON DIFFERENT WOODY SPECIES
Cristina Rodica MĂNESCU, Claudiu BUDA, Florin TOMA, Sorina PETRA

Abstract
The most known spontaneous species of woody climber, common ivy, it was found to prefer some species more then
others as host for its growth and development. Different trees and shrubs from the biggest urban park in Bucharest
were investigated to find the presence of spontaneous growth of Hedera helix on their trunks and branches. Seven
species of trees and three shrubs were labeled as preferred hosts (100% presence) for Hedera helix, no matter their age
or health condition, while four species of trees and nine shrubs were found as totally inconvenient (0% presence) for ivy
growth. It was concluded that some species of woody plants create better conditions for Hedera helix to grow as an
invasive plant, with considerable repercussions on the plantations management programs.
Key words: Hedera helix, host species, invasive growth, plantations management.
INTRODUCTION various birds, which consume the fruits

(Mitchell, 1975; Reichard, 2000). Then, the ivy
Ivy (Hedera helix L.) is a natural presence in grows covering the ground and climbing the
the European temperate forests, especially trees, in some cases became invasive
broadleaves ones (Rizzetto et al., 2016; Moser (Beekman, 1984; Trémoliéres et al., 1998;
et al., 2017). Unlike other lianas species of Badre et al., 1998; Schnitzler and Heuzé,
these habitats, such as Clematis vitalba or 2006). On some species of trees Hedera may
Lonicera caprifolium, ivy can grow at the add weight and increase the storm damage
ground level, covering the soil, or at the canopy (Reichard, 2000; Schnitzler and Heuzé, 2006).
layer, climbing the trees. Thus, Hedera species Invasive growth necessitates costly eradication
were considered having a positive impact on programs in some countries.
the forest, in general, but mainly on the host The objectives of this work were to examine
tree, because of nutrients inputs in spring with ivy population dynamics in forest-like
the foliage fall, tree stability and attracting and plantations of one of the biggest urban park in
hosting desirable organisms (Trémoliéres et al., Europe and provide information about host
1988; Bell et al., 2012; Smets et al., 2016; species that ivy prefers to attach and develop.
Ruggeri et al., 2016).
In urban environment, green areas are made by MATERIALS AND METHODS
a mixture of native and exotic species, for
ornamental reasons but also as a consequence The study was conducted in Herăstrău Park,
of preventing the cities problems such as situated in the northern side of Bucharest,
pollution or poor soils (Barrico et al., 2018; Romania. Opened in 1936, the park covers an
Müller et al., 2018; Vieira et al., 2018). area of 187 ha and is bordering by high traffic
Furthermore, the remaining urban forests were streets. For this reason, the peripheral limits of
sometimes completed with exotic species of the park were designed as a protective zone,
trees and shrubs. However, some of the native made from massive trees and shrubs
species, such as Hedera helix shows local plantations.
invasions in these plantations, especially when In these plantations, various species, mainly
the human interference is absent. Originally, broadleaves, both native and exotic, grow
ivy appears here from the seeds brought by together without any human intervention, as a
597
natural forest. The structure of these plantations that on young host trees the climbing behaviour
is characterized by medium-low dense woody of ivy was stronger in exotic species (68% at
vegetation, developed in three levels: shrubs, broadleaves species and 100% conifers). From
small trees and medium-big trees. these non-native species, we remarked ivy
We analysed in this study the ivy plants, from climbing more often: Acer negundo, Celtis
natural spreading, which appear frequently occidentalis, Robinia pseudacacia, Thuja
covering the ground or the plants (both trees orientalis and Pinus nigra.
and shrubs) in these massive plantations.
Area of study was limited at the south of the dbh values (cm)
park (approximate 20 ha), where ivy is much 90-99
more present, growing frequently on trunks and 80-89
70-79
branches of trees and shrubs. 60-69
For evaluation of ivy preference for certain 50-59
conifers
hosts, the proportion of invaded hosts and 40-49
broadleaves
condition of the hosts (Table 1), this area was 30-39
20-29
divided in plots. In each plots, all shrubs and 10-19
trees of >5 cm diameter at breast height (dbh) 5-9
were inventoried, identified and analysed for %
0 20 40 60
age, health condition and ivy presence.
Figure 1. Frequency of ivy on different dbh of host trees
Table 1. Category of woody plants condition
Class Condition Description
A Excellent Healthy, vigorous plants
Independent of species, the diameters of large
Healthy, vigorous plants in general, host tress were in positive correlation with
B Medium
but with maximum 25% wounds or the height of ivy on trunks (Pearson’s R = 0.54,
dead branches. No disease or
parasitic attack. P < 0.001). The occurrence of ivy per layers of
Plants vigour affected. Unhealed vegetation was considerably higher at shrub layer
C Poor wounds or chronic parasitic attack to (Table 2).
maximum 50%.
Plants with large dead branches,
Irreversible cavities or signs of internal decay to
Table 2. Distribution of ivy per layers of vegetation
D
decline maximum 75%. Irremediable Shrub layer Subcanopy Canopy
damaged.
(1.5-5m) (5-15 m) (>15m)
E Dead -
Mean density of host
plants (per ha) in 5.9 18.9 11.6
different layers
Observations and measurements were carried
Occurrence of ivy per
out during the autumn-winter in 2016 and layers of vegetation 62 34 4
2017. The relationship between quantitative (%)
data was examined using statistical analysis
(Pearson’s R). Although subcanopy is much more represented
in the massive plantations of this park, ivy
RESULTS AND DISCUSSIONS grows and develops better below these,
particularly because of the light conditions
Ivy was found frequently growing on trees offered at this level. Most of the trees growing
located at the edge of the tree massive, near in the subcanopy have small or medium leaves
main alleys or lawns. In some zones of the (57%, respectively 23%) and low density
massive plantations, ivy was present only at the crowns.
ground level. However, we establish a mean Ivy proved to choose certain species as hosts
density of ivy hosted by shrubs and trees of (Figure 2). According with the ivy preference,
36.4 ivies per hectare of green area. four groups of host species were identified: the
The host trees dbh values indicated that ivy was most attractive (100% of the trees support
more present on mature trees (Figure 1). The ivies), highly attractive (>60% of the trees
number of host trees with small dbh values was support ivies), attractive (20-50% of the trees
expected to be lower, because of climbing of support ivies) and less attractive (<20% of the
ivy on particular barks. Anyway, we observed trees support ivies).
598
veness (Figure 3). The majority of shrubs
species were unattractive for ivy. In this case,
Thuja orientalis
the most attractive

Pinus sylvestris
Tilia platyphyllos even the ivy was present at the ground level or
Populus nigra
Gleditsia triacanthos in trees nearby shrubs (in different light con-
Carpinus betulus
Acer tataricum
ditions) it was not covering at all their base or
Prunus cerasus branches. However, three of them - Corylus
Pinus nigra
Acer negundo
avellana, Laburnum anagyroides and
Robinia pseudacacia Sambucus nigra, all native species with
Fraxinus excelsior
Sophora japonica
vigorously growing, were highly preferred by
Catalpa bignonioides ivy.
Quercus robur
Acer platanoides The evaluation of hosts’ condition showed a
Picea pungens
Abies concolor
tendency of ivy to climb trees in poor and
Ulmus carpinifolia irreversible decline more than the others (Table
Celtic occidentalis
Taxus baccata
3).
Tilia tomentosa
Table 3. State of vegetation of the host trees (%)
Malus sylvestris unattractive
Fagus sylvatica
Class Class Class Class
0 20 40 60 80 100 A B C D
Percentage of host tree Broadleaves species 14.5 20.8 20.8 43.7
Conifers species 11.1 33.3 55.5 -
Figure 2. Preference of ivy for host tree species
Correlation between trees’ state of vegetation

Seven species of trees are the most preferred by
and the incidence of ivy on their trunk showed
ivy: Acer tataricum, Carpinus betulus,
that ivies are significantly more present in trees
Gleditsia triacanthos, Populus nigra, Tilia
with poor condition (Pearson’s R = 0.63, P <
platyphyllos, Pinus sylvestris and Thuja
0.001).
orientalis. Other eight tree species were found
Still, in some species, such as: Acer negundo,
highly attractive for ivy, most of them exotic
Acer tataricum, Carpinus betulus, Catalpa
and much appreciated for their biological and
bignonioides, Fraxinus excelsior, Gleditsia
ecological characteristics for this type of
triacanthos and Robinia pseudacacia, even
plantations. Four species (all native) proved to
with a very good condition of trees (class A),
be unattractive for ivy: Fagus sylvatica, Malus
ivy was found growing on their trunk. All of
sylvestris, Tilia tomentosa and Taxus baccata.
these species develop a rough bark since early
Sambucus nigra
stages of growth. These results confirm some
other studies (Hegarty and Caballe, 1991;
the most attractive
Laburnum anagyroides
Corylus avellana Schnitzler and Heuzé, 2006; Leicht-Young et
Lonicera tatarica
Syringa vulgaris
al., 2010; Steinbrecher et al., 2010), which
Philadelphus coronarius demonstrated that ivy, like other lianas, prefer
Symporicarpos albos
Spirea x vanhouttei
the rough barks for support.
Rhus typhina The tendency of invasive growth of ivy was
observed at some species (Figure 4). A
unattractive
Hydrangea macrophilla
proportion of coverage over 60% of the total

Hibiscus syriacus
Forsythia x intermedia
Deutzia scabra surface of host plant was remarked at 15
Crataegus monogyna
Chaenomeles japonica
different species of trees and shrubs. Over 60%
0 20 40 60 80 100
from these are exotic species and commonly
Percentage of host shrubs
present in urban green spaces.
Values of host coverage were extremely high
Figure 3. Preference of ivy (85%) at some species: Catalpa bignonioides,
for host shrubs species Fraxinus excelsior, Gleditsia triacanthos,
Lonicera tatarica, Quercus rubra, Robinia
For the shrubs placed near or in the massive pseudacacia, Sambucus nigra, Syringa vulgaris
tree plantations, most of them exotic species and Thuja orientalis.
(65%), ivy proved also a different attracti-
599
Mediterranean climates. ISHS 2010, Acta Hort.,
Tilia platyphyllos
Thuja orientalis Landscape & Urban Hort., 937, 1033-1040.
Syringa vulgaris Hegarty E., Caballe G., 1991. Distribution and
Sophora japonica abundance of vines in forest communities. In: Putz
Sambucus nigra
Robinia pseudacacia
F.E., Mooney H.A. (Eds.), The Biology of Vines.
Quercus rubra Cambridge University Press, Cambridge, 313-337.
Pinus nigra Leicht-Young S.A., Pavlovic N.B., Frohnapple K.J.,
Philadelphus coronarius
Lonicera tatarica
Grundel R., 2010. Liana habitat and host preferences
Gleditsia triacanthos in northern temperate forests. Forest Ecology and
Fraxinus excelsior Management 260, 1467-1477.
Catalpa bignonioides
Mitchell A.F., 1975. Three Forest Climbers: Ivy, Old
Acer platanoides
Abies concolor
Man’s Beard and Honeysuckle. Forestry Commission
0 20 40 60 80
Forest Record 102, H.M.S.O., London.
Moser B., Büntgen U., Molinier V., Peter M., Sproll L.,
Maximum coverage of host (%)
Strobbe U., Tegel W., Egli S., 2017. Ecological
indicators of Tuber aestivum habitats in temperate
Figure 4. Percentage of ivy coverage at some species European beech forest. Fungal Ecology 29, 59-66.
Müller A., Bøcher P.K., Fischer C., Svenning J.C., 2018.
The invasive growth of ivy is rare in urban ‘Wild’ in the city context: Do relative wild areas
plantations, but it was reported by some authors offer opportunities for urban biodiversity? Landscape
and Urban Planning, vol. 170, 256-265.
for ivies growing in natural forest (Schnitzler Reichard S., 2000. Hedera helix L. In: Invasive plants of
and Heuzé, 2006; Rizzetto et al., 2016). California’s wildlans. Berkeley, USA: University of
California Press, 212-216.
CONCLUSIONS Rizzetto S., Belyazid S., Gégout J.C., Nicolas M., Alard
D., Corcket E., Gaudio N., Sverdrup H., Probst A.,
2016. Modelling the impact of climate change and
Urban massive trees and shrubs plantations are atmospheric N deposition on French forests
made of native and exotic species. Ivy coming biodiversity. Environ. Pollution 213, 1016-1027.
from natural sites can appear and populate Ruggeri R., Provenzano M.E., Rossini F., 2016. Effect of
dense plantations of green areas in certain mulch on initial coverage of four groundcover
conditions. Our results showed that some of the species for low input landscaping in a Mediterranean
climate. Urban Forestry & Urban Greening 19, 176-
exotic species, especially trees, are more 183.
susceptible as host for ivy. In this case, Schnitzler A., Heuzé P., 2006. Ivy (Hedera helix L.)
maintenance of massive plantations can require dynamics in riverine forests: Effects of river
further attention and cost more. For this reason, regulation and forest disturbance. Forest Ecology and
selection of unattractive or less attractive Management 236, 12-17.
Smets W., Wuyts K., Oerlemans E., Wuyts S., Denys S.,
species for ivy or even reducing the proportion Samson R., Lebeer S., 2016. Impact of urban land use
of attractive host species may be a future way on the bacterial phyllosphere of ivy (Hedera sp.).
to solve the problem of managing ivy Atmospheric Environment 147, 376-383.
propagation in urban plantations. Steinbrecher T., Danninger E., Harder D., Speck T.,
Kraft O., 2010. Quantifying the attachment strenght
of climbing plants. Acta Biomaterialia 6, 1497-1504.
REFERENCES Trémoliéres M., Carbiener R., Exinger A., Turlot J.C.,
1988. Un exemple d’interaction non compétitive
Badre B., Nobelis P., Trémoliéres M., 1998. Quantitative entre espèces ligneuses: le cas du lierre arborescent
study and modelling of the litter decomposition in a (Hedera helix L.) dans la forèt alluviale. Acta Oecol.
European alluvial forest. Is there an influence of 9, 187-209.
overstorey tree species on the decomposition of ivy Trémoliéres M., Sanchez-Perez J.M., Schnitzler A.,
litter (Hedera helix L.)? Acta Oecol. 19 (6), 491-500. Schmitt D., 1998. Impact of river management
Barrico L., Castro H., Coutinho A.P., Gonҫalves M.T., history on the community structure, species
Freitas H., Castro P., 2018. Plant and microbial composition and nutrient status in the Rhine alluvial
biodiversity in urban forests and public gardens: hardwood forest. Plant Ecology 135, 59-78.
Insights for cities’ sustainable development. Urban Vieira J., Matos P., Mexia T., Silva P., Lopes N., Freitas
Forestry & Urban Greening 29, 19-27. C., Correia O., Santos-Reis M., Branquinho C., Pinho
Beeckman F., 1984. La dynamique d’une forèt alluviale P., 2018. Green spaces are not all the same for the
rhénane et le rôle des lianes. In: Cramer, J. (Ed.), Les provision of air purification and climate regulation
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Strasbourg, 475-501. Research, vol. 160, 306-313.
Bell N.C., Owen J., Altland J., 2012. Selection of
groundcovers for low-input landscapes in temperate
600
POLLEN ANALYSIS IN SOME TULIP CULTIVARS
Mirela Irina CORDEA1, 2, Ioana Delia POP1, 2, Sonia BORS-OPRIŞA1, 2,

Lucia MIHALESCU3
1
Faculty of Horticulture, 3-5 Mănăştur Street, 400372, Cluj-Napoca, Romania
2
Advanced Horticultural Research Institute of Transylvania, 3-5 Mănăștur Street, 400372,
Cluj-Napoca, Romania
3
Technical University of Cluj-Napoca, North University Center of Baia Mare, Romania
Abstract
Viability and germination capacity of pollen grains is very important in breeding programs which require a minimum
30% germination level for the success of artificial hybridization. This paper presents the pollen viability and
germination capacity analyses performed in nine tulip cultivars that belong to six different groups and the correlation
between these two determinations. The viability of the pollen grains registered higher values in eight, out of nine
cultivars (ʻYokohamaʼ - 81.7%, ʻVeronique Sansonʼ - 93.9%, ʻTender Whisperʼ - 90.7%, ʻPaul Schererʼ - 86.0%,
ʻDavenportʼ - 84.8%, ʻFancy Frillsʼ - 92.9%, ʻBlue Parrotʼ - 96.1%, ʻRed Riding Hoodʼ - 89.8%), while in the case of
germination capacity only four cultivars (ʻRed Riding Hoodʼ - 35.9%, ʻBlue Parrotʼ - 81.1%, ʻDavenportʼ - 78.3% and
ʻSalmon Impressionʼ - 27.7%) registered the necessary rate for the breeding programs. The lower germination
percentage of pollen grains, in some cultivars, may be due to the duration of storage period of the pollen.
Key words: breeding, pollen grain germination, pollen grain viability, tulips.
INTRODUCTION cultivars belonging to six classification groups

in order to be used in breeding programs.
Although there are thousands of tulip cultivars
over the world, breeders do not cease studying MATERIALS AND METHODS
and creating new ones in order to satisfy the
need for beauty of all mankind. Tulips, along Nine tulip cultivars belonging to six classi-
with daffodils, hyacinths, gladioli are the most fication groups were tested in order to deter-
widely cultivated bulbous spring plants used to mine the germination capacity and viability of
decorate green areas and also as cut flower in pollen grains (Brickell and Zuk, 1997;
bouquets and other floral arrangements. Raamsdonk and de Vries, 1996) (Table 1), tests
The study of germination capacity of pollen having been performed in the Plant breeding
grains is very important in the selection of laboratory of the Advanced Horticultural
genitors and in a successful artificial hybridi- Research Institute of Transylvania, UASVM
zation, where its level should be at least 30% Cluj-Napoca.
(Cordea, 2014). Germination capacity of pollen
grains may be determined indirectly by testing Determination of germination
the pollen viability and directly, by pollen Germination capacity of pollen grains presumes
germination in vitro on solid nutrient medium. that the fertile grains with viable cytoplasm and
Viability of pollen grains may be revealed with spermatia germinate and release pollen tube on
simple colouring tests (with potassium iodide an artificial nutritional medium enhanced with
or carmine staining method). It is well known different sucrose concentrations (10-30%), with
that there is a very strong direct, positive 70-90% humidity at a temperature between
correlation between germination and viability 20-22°C. There are different opinions on the
of pollen grains (Cordea, 2014). concentration of the two ingredients which
The objective of this study was to evaluate the form the nutritional medium. Soares et al.
pollen germination and viability in nine tulip (2013) has obtained the best results on a
601
medium with 15% sucrose and 0.8% agar in
Passiflora sp. Some authors (Chagas et al.,
2008) observe that higher concentrations of
agar increase the germinated pollen grain ratio
in pear cultivars while others (Luza and Polito,
1985) came to the conclusion that in nut
cultivars a relative high concentration of agar
inhibits pollen germination and decreases the
pollen tube length.
Our tests were performed on solid nutritional
medium according to the protocol described by
Cordea (2014). In this regard anthers were Figure 1. Pollen grain germination in tulips -
collected from unopened flower buds before magnification 10x (filled arrow - germinated grains;
the pollen has reached maturity. The anthers dashed arrow - non-germinated grains) (source:original)
have been placed on Petri dishes and
maintained for a period of 48-72 hours in room Viability analysis
temperature (22oC) until they broke and Viability of pollen grains was examined by
released the pollen grains. The dried pollen means of the potassium iodide colouring test.
grains have been sown on solid medium made In the presence of the colouring agent fertile
of 1.5% agar and 15% sucrose with 85% grains with viable cytoplasm and spermatia will
humidity, preserved at 22oC temperature. colour in black/dark brown while the sterile
The microscope readings have been performed ones will remain colourless (Figure 2).
in 24 hours after sowing the pollen grains on
the nutritional medium. In order to obtain a
precise counting of the examined grains there
were chosen certain uncrowded microscopic
fields and observations have been made with
the 4x and 10x lenses (Figure 1).
Table 1. The tulips cultivars

at UASVM Cluj-Napoca
Flower
No. Cultivars Groups
colour
I – Single early
1 ʻYokohamaʼ Yellow
tulips
ʻVeronique III – Triumph Orange
2 Figure 2. Pollen grain viability in tulips: A -
Sansonʼ tulip with red
magnification 10x; B - magnification 40x (filled arrow -
ʻTender III – Triumph Fuchsia viable grains; dashed arrow - non-viable grains)
3
Whisperʼ tulip with white (source:original)
ʻPaul III – Triumph Dark
4
Schererʼ tulip purple In this purpose, anthers were sunken imme-
ʻSalmon IV – Darwin Light pink diately after collecting in the Carnoy fixative
5
Impressionʼ hybrid tulips salmon
solution for a period of 1.5 hours then passed
VII – Fringed Red with
6 ʻDavenportʼ into alcohol until determination.
tulips yellow
For high accuracy, in both determinations there
ʻFancy VII – Fringed
7
Frillsʼ tulips
Pink were analysed about 300 pollen grains in
ʻBlue
several microscopic fields, then the rate (%) of
8 X – Parrot tulips Violet germination and viability was calculated.
Parrotʼ
ʻRed Riding XIV – Greigii
9 Red
Hoodʼ tulips
602
RESULTS AND DISCUSSIONS minimum 30%. The same situation is observed
in ʻYokohamaʼ and ʻFancy Frillsʼ cultivars.
In plant breeding the artificial cross pollination
is a very important proceeding in creation of
genetic variability, which will then be exploited 120
Germination and viability level (%)

93,9 90,7 96,1
by selection in order to obtain new cultivars 100 86 84,8 92,9 89,8
81,7 78,3 81,1
with higher ornamental value than those on the 80
market. 60
In tulip species there are multiple incompati- 35,9
40 27,7
bility barriers in interspecific hybridization as 14,3 13,1
23,2
20 8,1 7,8 11
described by Kho and Baër (1971). To achieve
a successful artificial hybridization, pollen 0
grains should have a minimum 30% germi-
nation rate, very important to be taken in con-
sideration when choosing the genitors in plant
breeding programs.
The analysis of pollen grains of each cultivar of
our tulip collection is presented in Figure 3. Germinability Viability
Examining the results obtained there can be
Figure 3. Pollen grain germination and viability in nine
noticed that the highest germination capacity tulip cultivars, UASVM Cluj-Napoca, 2017
rate has been registered in ʻBlue Parrotʼ
cultivar (81.1%) while the lowest in ʻVeronique
Sansonʼ (7.8%). A special behaviour presents the ʻSalmon
It is also noticeable that only four cultivars Impressionʼ cultivar (IV - Darwin hybrid tulip
(ʻBlue Parrotʼ - 81.1%, ʻDavenportʼ - 78.3%, group) which encountered a rather low rate
ʻRed Riding Hoodʼ - 35.9%, ʻSalmon both in germination and viability.
Impressionʼ - 27.7%) have exceeded the 30% According to the data presented in Figure 3 it
germination rate, the minimum level for a can be noticed that the viability of pollen grains
successful artificial hybridization. in all tested cultivars reached much higher
Within the groups germination capacity varies levels than their germination rate, except the
as it follows: all the three cultivars belonging to ʻSalmon Impressionʼ.
group III - Triumph tulip, presented very low
germination rate (ʻVeronique Sansonʼ - 7.8%; CONCLUSIONS
ʻTender Whisperʼ - 14.3% and ʻPaul Schererʼ -
11.0%) while in group VII - Fringed tulips - Based on these results there has been reached
results are different. ʻDevenportʼ records a the conclusion that displaying a higher viability
rather high germination rate (78.3%) while in than germination rate might be due to the
ʻFancy Frillsʼ only 23.3% of the pollen grains length of the drying and preservation period of
germinated. This denotes that in cultivars of the pollen between collecting and analysis (4-5
same group pollen grains do not necessarily days). This suggests that the fertile period of
germinate in the same rate. tulip pollen is rather short, only a few days.
As far as viability is concerned data presented As a consequence of the pollen viability and
in Figure 3 reveal a higher rate than germi- germination analysis in our tulip collection
nation. Almost all cultivars, regardless the there can be concluded that only three cultivars
group they belong to, present a rather high (ʻDavenportʼ, ʻBlue Parrotʼ and ʻRed Riding
viability rate between 81.7% (ʻYokohamaʼ) and Hoodʼ) presented fairly high rates in both
96.1% (ʻBlue Parrotʼ). determinations, while the ʻRed Riding Hoodʼ
Cultivars of group III - Triumph, present a high cultivar slightly passed the required
viability level (ʻVeronique Sansonʼ - 93.9%; germination level. These results offer important
ʻTender Whisperʼ - 90.7% and ʻPaul Schererʼ - information for selecting the best female/male
86.0%) as compared to their germination rate genitor in tulip breeding programs for higher
which does not succeed to reach the necessary ornamental value.
603
REFERENCES Hernández Bermejo J.E., García Sánchez E., 2009.
Tulips: An Ornamental Crop in the Andalusian
Beyhan N., Serdar U., 2008. Assesment of pollen Middle Ages1 Economic Botany, 63 (1), 60-66.
viability and germinability in some European chesnut Kho Y.O., Baër, 1971. Incompatibility problems in
genotypes (Castanea sativa L.). Hort. Sci. (Prague), species crosses of tulips, Euphytica 20, 30-35.
35 (4), 171-178. Luza J.G., Polito V.S., 1985. In vitro germination and
Brickell C, Judith D. Zuk., 1997. The American storage of English walnut pollen. Scientia
Horticultural Society A-Z encyclopedia of garden Horticulturae, 27, 303-316.
plants. New York. Okazaki K., 2005. New Aspects of Tulip Breeding:
Chagas E.A., Barbosa W., Saito A., Pio R., Chagas P.C., Embryo Culture and Polyploid, Acta Hort. 673 (127-
2008. In vitro germination of Pyrus calleryana Decne. 140).
pollen: adjusting a protocol. Acta Horticulturae, 800, Raamsdonk L.W.D. van, T. de Vries, 1996. Cultivar
515-520. classification in Tulipa L. (Liliaceae). Acta Bot.
Cristea V., 2014. Vascular plants: diversity, systematic, Neerl. 45: 183-198.
ecology and importance. Ed. Cluj University Press, Radović A., Nikolić D., Milatović D., Đurović D.,
Cluj-Napoca. Trajković J., 2016. Investigation of pollen morpholo-
Cordea M.I., 2014. Plant Breeding - practical works. gical characteristics in some quince (Cydonia oblonga
AcademicPres, Cluj-Napoca, Romania. Mill.) cultivars. Turk J Agric For 40: 441-449.
De Assis Sinimbú Neto F., Martins A.B.G., Barbosa J.C., Van Tuyl J. M., Van Creij M.G.M., 2006. Tulip: Tulipa
2011. In vitro viability of ‘bacury’ pollen grains, gesneriana and Tulipa hybrids. In: Flower Breeding
Revista Brasileira de Fruticultura, vol. 33 (2), 593- and Genetics - Issues, Challenges and Opportunities
600. for the 21st Century (ed. Anderson N.O.), Ed.
Springer.
604
THE EFFECT OF DIFFERENT LIGHT CONDITIONS

ON THE GROWTH AND DEVELOPMENT OF CHLOROPHYTUM
AMANIENSE ENGL. ‘FIRE FLASH’
Manuela MANDA1, Carmen NICU1, Diana ZAMFIR-VÂȘCĂ2

1
University of Craiova, Faculty of Horticulture, 13 Al.I. Cuza Street, Craiova, Dolj, Romania
2
Abstract
Chlorophytum amaniense Engl. is a foliage plant, member of the family Liliaceae, originates from the rainforests of
East Africa. ‘Fire Flash’, the only cultivar of this species, known by several common names including Fire Glory,
Mandarin Plant and Tangerine, present unique decorative characteristics. C. amaniense‚ ʻFire Flash’ do not support
the direct action of the sun's rays and the placement in bright exposures. Given the claims of C. amaniense versus light
and insufficient information in the literature, the objective of this research was to study the reaction of ‘Fire Flash’
under different lighting conditions. Plants attained the greatest leaf size and petioles when ‚ʻFire flash’ were grown in
low and medium light intensity while plants exposed to high light showed lowest values of growth parameters. In
addition, high light intensity produced chlorosiss and leaf burn and plants have become unmarketable after a 3 months
period. The best size and quality of ‘Fire Flesh’ plants, occurred when plants were grown at a medium light levels of
80 μmol m-2 s-1.
Key words: Chlorophytum amaniense, growth, light, photosynthetic parameters.
INTRODUCTION Unlike C. comosum (Thunb.) Jacques (spider

plant), C. amaniense ‘Fire Flash’ does not
The genus Chlorophytum belongs to Liliaccae produce stolons, and propagation could be
and encompasses about 200 species chiefly realized through seed, division and in vitro
native to tropical Africa, Australia and Asia regeneration (Cui et al., 2011).
(Anton, 2009). Most species are evergreen In addition to ornamental value, the rhizomes
perennials with rhizomatous roots either short of ‘Fire Flash’ form nearly oval tubers, which
and fibrous or thick and translucent. may contain antitumor steroidal saponins as do
Chlorophytum amaniense Engl., member of the other species of Chlorophytum Ker. Gawl
family Liliaceae, is a foliage plant originates (Kaushik, 2005; Cui et al., 2011). Of the twelve
from the rainforests of East Africa. ‘Fire Flash’, species analyzed, C. amaniense Engl., showed
the only cultivar of this species, known by highest phenol, flavonoid, saponin contents and
several common names including Fire Glory, in vitro antioxidant activity (Shinde et al.,
Mandarin Plant and Tangerine, present unique 2016; Patil, 2016).
decorative characteristics (Anton Doina et al., ‘Fire Flash’ is grown in shady greenhouses at
2006). temperatures between 18-32°C (optimum
Bright coral petioles and midveins contrast temperature between 24-29°C) and relative
with deep green, ovate-lanceolate leaves humidity between 50% and 100%. Water used
making an exotic appearing and exciting plant. for ‘Fire Flash’ production should be free of
The flowers are white in a dense cylindrical fluoride since it may cause leaf necrosis. Light
panicle partially hidden by the foliage. intensity is extremely important for quality of
The inflorescence does not add any value to the ‘’Fire flash” plant production (Chen et al.,
aesthetic appearance. In fact, it is detrimental. 2002). C. amaniense ʻFire Flash’do not support
Removal of the inflorescence at an early stage the direct action of the sun's rays and the
of flowering improves plant growth (Chen et placement in bright exposures. Depending on
al., 2002). the intensity of light, the leaves may have
605
different shades from intense green to light 10 hours per day and were maintained in
green. It also influences the color and length of moderate shade during the first month and then
petioles, these being the main decorative transferred to the different light treatments (in
element. Higher light levels will cause leaf the first week of March 2016), by placing the
chlorosis and, eventually, scorching and plants in three areas of the greenhouse with
necrotic lesions that can cause unsalable plants. different light level: low light intensity (LL):
Based on the evaluation study, Chen et al. 7.98-9.12 μmol·m−2 s−1; medium light intensity
(2005 a) recommended that ʻFire Flash’ can be (ML): 77.9-82.46 μmol·m−2 s−1; high light
propagated through seed, division, or tissue intensity (HL): 220.4-233.7 μmol·m−2 s−1.
culture and produced as a potted foliage plant The light intensity inside the greenhouse was
under light levels from 114 to 228 μmol m-2 s-1 measured with a Lux meter at 10.30 am, 2.00
and temperatures from 18 to 32°C. After being pm and 4.00 pm, in three zone of greenhouse
placed in building interiors, plants should be with different light levels. These evaluations
located in interior light levels between 50 to were performed on fivedifferent days in an
200 foot candles (10-40 μmol m-2 s-1) (Chen et interval of five months corresponding to the
al., 2005 b). ‘Fire Flash’ plants are able to experimental time. The luminous flux per unit
maintain their aesthetic appearances under a area was converted into photon flux density
low light level of 8 μmol m−2 s−1 for 8 months (PFD) using a conversion factor of 0.0185
or longer (Chen et al., 2005 a). μmol photons m−2 s−1 per lux valid for sunlight
Given the claims of C. amaniense ‘Fire Flash’ (Hershey D.R., 1991; Pedersen et al., 2016).
versus light and insufficient information in the The experiment was conducted in a completely
literature, the objective of this research was to randomized design, using a monofactorial
investigate the response of C. amaniense plants arrangement with three treatments represented
under different light intensities. For this aimes by three levels of light availability (T1-LL; T2-
a pot experiment was conducted to analyze ML; T3-HL), with three replicates, and five
growth parameters and some photosynthetic plant per experimental unit.
parameters. The observations on average height of the
plants, number of leaves per plant, leaves size,
MATERIALS AND METHODS lenght of petiole, width of petiole were
recorded 90 days after the experiment was
The experiment was established at the Floricul- established. We also measured some photosyn-
ture Research Area, Faculty of Horticulture thetic parameters: the incident radiation in the
from Craiova (Romania), during February to leaf (Qleaf) expressed in μmol photons m-2 s-1;
August, 2016. The biological material consists stomatal conductance (gs), mol H2O m-2 s-1,
of plants of Chlorophytum amaniense Engl. transpiration (E), mmol H2O m-2 s-1, net
ʻFire Flash’ from the didactic greenhouse of the photosynthesis (A), μmol CO2 m-2 s-1, on the
Floriculture discipline. third or fourth leaf counting from the apex to
Ambient temperatures ranged from 20○C-22○C the base, by using Lcpro+® Portable
and relative humidity from 60%-80%. Photosynthesis System. Water Use Efficiency
We studied the effect of different lighting (WUE; µmol CO2 mmol–1 H2O) was calculated
intensity of the ʻFire Flash’ plants to assess the as A to E ratio (Ribeiro et al., 2009). The data
effect on morphological, ornamental and some were submitted to variance analysis and the
photosynthetic characteristics. averages compared by Tukey test at 5% error
Young plants of C. Amaniense ʻFire Flash’ probability (p <0.05) in MINITAB 16 software.
(obtained from seeds), with 4-5 leaves, were
selected and transplanted in the first week of RESULTS AND DISCUSSIONS
February 2016 into black, 2.8 liter pots (17 cm
diameter) filled with a substrate consisting of a Effects of Light Intensity on Plant Growth
40:30:30 mix of peat, coconut fibre, and perlite. In ornamental horticulture, the leaf forms and
Controlled-release fertilizers (Osmocote 18-6- color, sizes, and shapes of pot plants is
12) was added to the substrate. Plants were essential components of their visual quality that
placed in a greenhouse with natural light for 8- determines the commercial value of the
606
products (Boumaza et al., 2010). Light intensity species, there were also significant differences
influence plant characteristics and quality according to the intensity of the light. The
attributes (Runkle, 2013). Knowledge of the average length of petioles recorded the lowest
morphological and physiological characteristics value at HL (3 cm), and the highest value was
of C. amaniense in response to various light recorded at LL (6.8 cm). Instead, average
conditions is still sparse. Clear external diffe- petiole widths were between 1.2 cm in low
rences were observed among plants grown light intensity (LL) plants and 1.7 cm in
under five months under different light intensive light (HL) plants.
intensities. In addition to the measurable parameters, the
Regarding the average height of the plants, the study of C. amaniense ʻFire Flash’ plants under
highest values were recorded in an exhibition different illumination conditions had the fol-
characterized by low light intensity (LL-13,7 lowing results: low intensity (7.98-9.12 μmol
cm). Moreover, the plants grown in a high m−2s−1) caused a more intense color of the
intensity area of light recorded the lowest leaves and the petiole, the elongation and
average height (HL-9.7 cm). Similarly, shade thinning of the main decorative element, the
induced more growth in height in Passiflora petiole; high intensity (220.4-233.7 μmol
edulis f. flavicarpa plants (Valladares et al., m−2s−1) produced chlorosiss and leaf burn and
2000; Zanella et al., 2006). Apical dominance plants have become unmarketable after a 3
tends to increase when plants are submitted to months period. Visual observations indicated
high shade levels, due to a decrease in the that C. amaniense produced commercially
production of photoassimilates and the highest acceptable plants at 7.98-9.12 μmol m−2 s−1,
level of auxin at the stem apex bud (Vanneste however the optimal growth and development
and Friml, 2009; Woodward and Bartel, 2005) occurred from 77.9-82.46 μmol m−2 s−1.
The average leaf number on the plant was Although Chen et al. (2005 a), recommends as
maximum under medium illumination (ML - optimal light interval 10-40 μmol m−2 s−1 for
9.7), and in variants grown under low or high ʻFire Flash’ production, our results suggest that
light conditions, the values of this parameter a light level of about 80 μmol m−2 s−1 appeared
were significantly reduced (LL - 8.3 leaves, to be optimal.
respectively HL-7.3 leaves) (Table 1).
Placement of plants in areas with different Effects of light intensity on plant
luminous intensities caused significant diffe- photosynthetic parameters
rences in the average leaf length. Compared to Physiologically, light has both direct and
plants grown in intense light, where the average indirect effects. It affects on metabolism
value of this parameter was minimal (HL-21.5 directly through photosynthesis, whilst
cm), the plants placed in a low light intensity indirectly through growth and development
recorded the highest value (LL-32.5 cm), (Zhang et al., 2011). Table 2 presents the
followed at a significant difference by the effects of different light intensities on leaf
variant in which the plants received a medium photosynthetic parameters.
light intensity (ML - 27.2 cm). In contrast, the Net CO2 assimilation (A) was highest in HL
increase of the leaf width was proportional to (7.73 µmol CO2 m-2 s-1) followed by ML (6.81
the decrease in the shade, with the highest µmol CO2 m-2 s-1) treatment and, finally, LL
average values occurring under high light treatment (5.44 µmol CO2 m-2 s-1). Similar
intensity (HL-5 cm) (table 1). The lowest value results were obtained on Dieffenbachia
corresponds to the plants grown in an area with longispatha and Camellia x williamsii
a minimum intensity of light (LL - 4.5 cm). (Skillman et al., 2005; Fini et al, 2010). The
Low light intensity may lead to increase in leaf authors have shown that that A was higher in
number and leaf size and these changes may full sun and mild shade plants if compared to
maximize the capture of available light to meet heavy shaded plants.
the demand for leaf photosynthesis (Steinger et The greatest leaf stomatal conductance (gs) was
al., 2003). observed under medium light intensity (ML-
Regarding the average dimensions of the 0.12 mol H2O m-2 s-1), and the lowest under low
petioles, the main decorative element of this light intensity (LL-0.03 mol H2O m-2 s-1).
607
Stomatal conductance (gs) was signifycantly (LL-7.61 μmol CO2 mmol–1 H2O) compared to
higher in ML than that of all other treatments. plants placed under medium and high intensity
As also reported in other works, plants grown (ML-4.05 μmol CO2 mmol–1 H2O, HL-4.46
at high light are characterized by greater μmol CO2 mmol–1 H2O). No significant
stomatal conductances than plants grown at low differences were observed between the ML and
light (Baroli et al., 2008; Niinemets et al., HL treatments (Table 2).
2006). Previous studies also showed Intercellular CO2 concentration (Ci) showed
thatchanges in photosynthesis and transpiration same evolution as WUE. Light reduction
were correlated with stomatal conductance resulted in significantly higher values of Ci
(Greer, 2012; Miyashita et al., 2012). (LL-439.33 ppm) (Table 2). Previous studies
Transpiration (E) was affected by different showed that, in some species, the intercellular
shading treatments and followed a similar CO2 concentration (Ci) declined with increases
pattern to A. Significant differences were with the increase of light intensity (e.g. Hanba
observed between the HL irradiance treatment et al., 2002; Oguchi et al., 2005).
and all other treatments. There were significant Our findings show the higher light levels have
differences on leaf transpiration rate (E) provoked severe leaf damage, characterized by
between the HL treatment (1.75 mmol H2O m-2 leafchlorosis or scorching. Huang et al., 2015
s-1) and all other treatments (0.71-1.68 mmol show that under high light condition, excess
H2O m-2 s-1). absorbed light energy can induce photoinhi-
Water use efficiency (WUE) varied signify- bition explaining why shade-established
cantly with light intensity in C. amaniense species cannot survive under high light.
plants. Significant differences were observed
between plants submitted to high shade levels
Table 1. Effect of different light conditions on mean vegetative growth parameters at the end of experiment
Height of Number Length Leaf area Length of Width of

Treatments plants (cm) of leaf (cm) of leaf (cm) (cm2) petiole (cm) petiole (cm)
LL 13.7a 8.3ab 32,5a 109,53a 6.8a 1,2b
ML 11.2b 9.7a 27,2b 95,1ab 4,5b 1,5ab
HL 9.7b 7.8b 21,5c 80,65b 3c 1,7a
Means comparison were done using Tukey’s test (p<0.05). For each variable lowercase letters indicate comparison
among treatments and uppercase ones comparison among species.
Table 2. Net photosynthetic rate (PN), stomatal conductance to water vapour (gs), leaf transpiration rate (E),
intercellular CO2 concentration (Ci) and water use efficiency (WUE) of C. amaniense leaves were subjected to different
levels of irradiance
WUE
A E Gs Ci
Treatments (μmol CO2
(µmol CO2 m-2 s-1) (mmol H2O m-2 s-1) (mol H2O m-2 s-1) (ppm)
mmol–1 H2O)
LL 5.44 b 0.71 c 0.03 c 439.33a 7.61a
ML 6.81 ab 1.68 b 0.12 a 354.33ab 4.05b
HL 7.73 a 1.75 a 0.08 b 320.33b 4.46b
Means comparison were done using Tukey’s test (p<0.05). For each variable lowercase letters indicate comparison
among treatments and uppercase ones comparison among species.
CONCLUSIONS Light intensity had different effects on C.

amaniense growth. The results showed that C.
The cultivation of C. amaniense ʻFire Flashʼ in amaniense attained greatest hight of plant,
Romania is almost nonexistent, though is a number of leaf and leaf size when cultivated
exciting ornamental foliage plant as a result of under low light intensity. Under high light
its unique coral-colored midribs and petioles intensity values of plants grown were the
and tolerance to interior low light levels. smallest. With the reduction in light intensity,
608
petioles have elongated by over 50% but their irrigated vineyard conditions. AoB Plants, 1: 1-13.
diameter has decreased significantly. Visual https://doi.org/10.1093/aobpla/pls009.
Hershey D.R., 1991. Plant Light Measurement and
observations indicated that C. amaniense Calculations. The American Biology Teacher, 53 (6):
produced commercially acceptable plants at 351-353.
7.98-9.12 μmol·m−2·s−1, however the optimal Huang W., Zhang S.B., Zhang J.L., Hu H., 2015. Photo-
growth and development occurred from 77.9- inhibition of photosystem I under high light in the
82,46 μmol·m−2·s−1. Plants exposed to full light shade-established tropical tree species
Psychotriarubra. Frontiers in plant science, 6: 801.
conditions (220.4-233.7 μmol·m−2·s−1) become Kaushik N., 2005. Saponins of Chlorophytum Species.
unmarketable within a 3 months period. Our Phytochem Reviews, 4(2): 191-196.
findings suggests that the net photosynthesis Miyashita K., Tanakamaru S., Maitani T., Kimura K.,
(A) was higher in high (HL) and medium light 2005. Recovery responses of photosynthesis,
(ML) and low light intensity (LL) can decrease transpiration, and stomatal conductance in kidney
bean following drought stress. Environmental and
photosynthesis. The results from this study experimental botany, 53 (2): 205-214.
show that a light level of about 80 μmol m−2 Niinemets Ü., Cescatti A., Rodeghiero M., Tosens T.,
s−1 appeared to be optimal for C. amaniense 2006. Complex adjustments of photosynthetic
when both morphological and physiological potentials and internal diffusion conductance to
performance were considered. current and previous light availabilities and leaf age in
Mediterranean evergreen species Quercus ilex. Plant,
Cell & Environment, 29: 1159-1178.
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as a constraint for photosynthetic acclimation:
Anton D., Nicu C., Mandă M., 2006. Floricultură differential responses in leaf anatomy to increasing
specială - vol. II, Editura Universitaria, Craiova. growth irradiance among three deciduous trees. Plant,
Boumaza R., Huché-Thélier L., Demotes-Mainard S., Le Cell & Environment, 28: 916–927.
Coz E., Leduc N., Pelleschi- Travier S., Qannari E.M., Patil S.M., Rane N.R., Adsul A.A., Gholave A.R., Yadav
Sakr S., Santagostini P., Symoneaux R., Guérin V., S.R., Jadhav J.P., Govindwar S.P., 2016. Study of
2010. Sensory profiles and preference analysis in molecular genetic diversity and evolutionary history
ornamental horticulture: the case of the rosebush. of medicinally important endangered genus
Food Quality and Preference, 21: 987-997. Chlorophytum using DNA barcodes. Biochemical
Chen J., Henny R.J., McConnell D.B., 2002. Systematics and Ecology, 65: 245-252.
Development of new foliage plant cultivars. In Janick, Pedersen O., Colmer T.D., Borum J., Zavala Perez A.,
J., and Whipkey, A. (eds). Trends in New Crops and Kendrick G.A., 2016. Heat stress of two tropical
New Uses. ASHS Press, Alexandra, 466-472. seagrass species during low tides-impact on
Chen J., McConnell D.B., Henny R.J., Everitt K., underwater net photosynthesis, dark respiration and
Caldwell R.D., 2005 a. ʻFire Flashʼ: An Exotic diel in situ internal aeration. New Phytologist, 210 (4):
Ornamental Foliage Plant, Hort Technology July- 1207-1218.
September, 15 (3): 686-689. Ribeiro R.V., Machado E.C., Santos M.G., Oliveira R.,
Chen J., McConnell D.B., Henny R.J., 2005 b. Cultural 2009. Photosynthesis and water relations of well-
Guidelines for Commercial Production of watered orange plants as affected by winter and
Interiorscape ʻFire Flashʼ (C. amaniense). summer conditions. Photosynthetica, 47 (2): 215-222.
Environmental Horticulture Department, Cooperative Runkle E.S., 2013. Manipulating Light Quality to Elicit
Extension Service, Institute of Food and Agricultural Desirable Plant Growth and Flowering Responses.
Sciences, University of Florida. IFAC Proceedings Volumes, 46 (4): 196-200.
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Cui Jin, Juanxu Liu, Jianjun Chen, Richard J. Henny, A.R., Pawar P.K., Govindwar S.P., 2016.
2011. Regeneration of Chlorophytum amaniense ‘Fire Comprehensive investigation of free radical
Flash’ Through Indirect Shoot Organogenesis, Hort quenching potential, total phenol, flavonoid and
Science, 46 (3): 466-469. saponincontent, and chemical profiles of twelve
Baroli I., Price, G.D., Badger M.R., von Caemmerer S., Chlorophytum Ker Gawl. species. Indian Journal of
2008. The contribution of photosynthesis to the red Natural Products and Resources (IJNPR) [Formerly
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Greer D.H., 2012. Modelling leaf photosynthetic and stressful environments II: adaptive value and costs of
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TINERETULUI PARK – FUNCTIONAL AND COMPOSITIONAL

COMPONENTS IN THE URBAN LANDSCAPE OF BUCHAREST
Anca STĂNESCU1, Suzana COCIOABĂ2

1
2
Ecological University of Bucharest, Faculty of Ecology and Environmental Protection,
1G Vasile Milea Blvd, Bucharest, Romania
Abstract
Tineretului Park is a component of the urban green spaces system of Bucharest, being one of the most important green
entities of the city. The current study highlights the diversity of the park landscape functions, according to the urban
needs regarding the population’s loisir activities. Furthermore, the aspects of the landscape composition are analyzed,
as well as the manner in which the socio-economic and urban development conditions have influenced the selection of
the location for the future park; moreover, the particular conditions of the site have had a determinant role in the
design of the actual landscape-architectural composition, as well in the options for choosing and distributing the loisir
functions in the whole park area.
Key words: urban park, landscape functions, landscape composition.
INTRODUCTION approach specific to the second half of the XX

century. The connections target three approach
Tineretului Park is located in the southern part levels for the design solution, which are
of Bucharest, the site being bordered by Şerban reciprocally determinant: physical, social -
Vodă Hill, Piscului Hill, Olteniţei Highway and functional and aesthetical. The physical level
Tineretului Boulevard. During the 1960’s the refers to the determination of the park’s
question was raised regarding the development location, which was defined by the following
of an urban park in this area, at that time the considerations:
location being referred to as “Plângerii Valley” - the necessity to develop the urban and green
(Marcus, 1958). The park’s surface is 83 ha, landscape towards the southern part of the city;
while the surface of the lake is 13 ha. - the nature of the soil in this area, which
This enclosure of the site has been envisaged in (except for Piscului Hill) did not allow the
the systematization and urban development development in normal economical conditions
plan of Bucharest since 1956; the park - as of any type of urban constructions;
distinct green entity - was to be part of large - the necessity to fill up the area of existing
green area (“feather”) in the southern part of urban green spaces at that date (Carol Park,
Bucharest. These aspects of urban development Bellu Cemetery) and the development - through
had been provided even since the Interbelic this operation of the green area mentioned
period (1935), when a sketch with general above;
development guidelines of Bucharest was - the construction of a valuable urban core as a
conceived (Marcus, 1958). point of perspective closure, which took place
along side the important urban artery - Base
MATERIALS AND METHODS Line North-South;
- highlighting the most favourable natural
The methods of research and evaluation of the elements: the difference of topographic level
green area “Tineretului” are based on between the cornice (high ground) and the
establishing connections between a functional Dâmboviţa Meadow, also the semi-circular
landscape approach (in which function is shape - amphitheatre style - of Piscului Hill and
predominant) and the compositional - stylistic Şerban Vodă Hill;
611
- eliminating an important harmful source in The style of the final composition (approved by
this area of the city, which was affected at that the beneficiary) is a mixt one, with a greater
time by the presence of swamps and trash percentage of the free-landscape style (Figure
deposited on the site, aspects which led to the 2). The complex parks of the 20th century find
formation of a pest hole (Marcus, 1958). their best compositional solution within the
The architectural-landscape composition was mixt style, which includes and combines in
founded on the concept of optimal exploitation resolution all the processing needs of different
of the natural configuration of the land. Four functions and areas of the park (Stănescu,
study versions for composition design were 2008).
created (Figure 1). Regarding composition, the designers develo-
ped a complex of access paths, axes and
perspective in tight correlation with the main
circulation arteries in the area, by which the
park is accessed. The access and the main axis
of the park lead to the Sala Polivalentă
building, where social cultural events are
hosted, which involve a large audience. Thus,
the most important alley is located alongside
the main axis, which is approximately 28
a) b) meters high (Figure 2).
The landscape architect chose the preservation
of the natural landscape, which is mostly the
optimal solution (Stănescu, 2008). However,
the terrain, being very degraded, required
ample remediation works in order to assure the
optimal levels of quality for the vegetation
development. In this context, approximately
290000 m3 of healthy soil were brought on site,
c) d) quantity mixed in with the existing soil. By
Figure 1. Study versions of Tineretului Park means of hydromechanical works, 236000 m3
of excavation and compensational fillings were
carried out.
In the place of the old swampland, a 13 ha lake
was created (Figure 2), which is supplied by
eight shafts of low depth (20 meters) upstream,
while the excess waters are evacuated to the
collector at the base of Piscului Hill.
The actual shape of the lake was created by
correcting an existing profile, while pursuing
the ample modification of the contour, which
was supposed integrated in the general compo-
sition by shape, sensibility planning solutions
conceived by the designer (Stănescu, 2008).
The park’s functionality is based on the terrain

conditions and the particularities of the site.
The landscape architect’s options for choosing
and distributing the loisir functions was
Figure 2. Final composition of Tineretului Park determined by the specifications of the project
theme and by the needs regarding the spending
612
of free time of the inhabitants. Regarding this, of an integrator concept in the creation of the
the functional aspects subscribed to the general designing solution, concept which brings
trends of urban landscape design in the 20th together at the same time physical, social –
century: the public gardens and parks have functional and aesthetic elements, all of which
developed in various directions in the 20th based on environmental factors, on the physical
century, while the dominant types were characteristics of the site, and also on the urban
represented by recreational and amusement development and socio-economic factors.
parks (Kluckert, 2005).
On the other hand, a garden should not be a
substitute for nature, but an artistic expression
for bringing the human being in close contact Figure 3. Diagram of the composition’s integrated
with nature - this aspect is met by a public park formation
because nature is used as a frame for relaxation
and fun (Kluckert, 2005). The synthesis of the determinant compositional
By applying the connections method in the characteristics of Tineretului Park is presented
compositional approach, the present study in Table 1.
demonstrates that Tineretului Park is the result
Table 1. Synthetic evaluation of the determinant compositional characteristics of Tineretului Park

GREEN AREA EVALUATION
CONNECTIVITY COMPONENTS EFFICIENCY IN THE
QUANTITY QUALITY
URBAN LANDSCAPE
PHYSICAL - urban definition - distance to city - personalization - urban comfort
- extension of the centre within the city’s - capitalization of
city’s green space - surface of the park: green space system natural potential
system 83 ha - urban landmark - accessibility
- area landscape - conformity with the - diversification of
- terrain site’s natural urban texture
composition character - beneficial effects in
the urban microclimate
FUNCTIONAL - integrator - landscape - coherent - social utility
AND SOCIAL character of functions: loisir, rest, organization of - diversification of free
functionality relaxation, sport, recreational activities time activities for the
- population and nautical recreation, - differential population
area serviced parade application for - multiple possibilities
- nature of use - facilities, equipment categories of users for socialization
(individuals or and physical - specificity of
groups) furnishing, distinct facilities in
- average use time for each type of proportion to the
(days/week) function park’s profile and
categories of users
AESTHETICAL - mixt composition - optimum - unity and variety - enrichment of the
type with mineral/vegetal ratio - conformism and general urban ambiance
predominant - shapes of water non-conformism - aesthetic diversity
elements from the bodies and alleys - diversity of mineral
free style - contours of and vegetal textures
- composition vegetation groups
principles: axis, and clusters
perspective,
symmetry and
asymmetry,
dominant,
composition centre,
contrast, accent,
calibration
The main function for Tineretului Park is that by the alleys system (including the lake’s
of rest - relaxation - promenade, materialized contour alley). Furthermore, the lake can be
613
used as an area for practicing nautical establishing at the same time a most valuable
recreation activities (boat rides). The park also natural support for the facilities corresponding
holds functions for practicing outside sports, to each type of function.
playgrounds for children, and also the rose
garden which corresponds to the functions of ACKNOWLEDGEMENTS
education and floral décor; the most diverse
functions are found within most of the contem- The current paper was elaborated on the project
porary European parks, e.g. Els Pinetons Park for Tineretului Park, developed in the design
from Ripollet del Valles - Spain. The project institute “Proiect Bucureşti” by architect
comprises the construction of a pedestrian alley Valentin Donose, the author of the project and
which surrounds the park and unites a series of the coordinator of the entire construction of the
platforms with the dimensions 8 × 55 m, having park.
various usages: barbeques, children’s games, a
bar for barbeque serving (Mostaedi, 2004). REFERENCES
CONCLUSIONS Kluckert E., 2005. European Garden Design. hf

ULLMANN - Tandem Verlag GmbH, Königswinter,
484.
The development of contemporary urban parks Mostaedi A., 2004. Landscape Design Today. Carles
answers to a wide range of functional and Broto & Josep Maria Minguet, Barcelona, 52.
aesthetical-compositional demands. All these Marcus R., 1958. Parcuri şi grădini în România. Editura
comply with the whole composition of the Tehnică, Bucuresti, 206.
park, spatial layout, the landscape functions Stănescu A., 2008. Peisagisică urbană vol. I. Editura
Printech, Bucuresti, 11-12, 50, 66.
included within the park, as well as with the Stănescu A., 2011. Peisagisică urbană vol. II. Editura
vegetation components and design. This aspect Printech, Bucuresti, 40-47.
affirms the landscape cogitation and also the
evolution in good condition of all the functions,
614
NATURAL HERITAGE CONSERVATION IN BUILT-UP AREAS.

STUDY ON THE CIRCUS PARK AS AN URBAN LANDSCAPE ASSET
Suzana COCIOABĂ1, Anca STĂNESCU2

1
Ecological University of Bucharest, Faculty of Ecology and Environmental Protection,
1G Vasile Milea Blvd, Bucharest, Romania
2
Abstract
Circus Park is one of the most distinguished green areas of Bucharest, being a veritable urban landscape resource.
Measuring 17 ha of land, it was designed during 1960-1961 following the plans of the architect Valentin Donose, who
formed and coordinated an interdisciplinary team within the Bucharest Project Institute, responsible with designing
green areas much needed to the capital at the time. This document aims to analyse the essential components which
form a park and bestow the attribute of an urban landscape resource: vegetation, water, alleys and ornamental
furniture. Circus park is the quintessential example which points to the idea that, just as man is attracted to water, he is
also instinctively attracted to trees and the spaces they compose. The distribution of foliage items contained in this
study cover a wide spectrum of trees, shrubs, lianas and herbaceous plants which range from wild to natural or man-
made architectural shapes. This inquiry recommends maintaining the concepts found in the initial plan of the park and
taking the necessary measures to preserve this natural heritage of Bucharest.
Key words: natural heritage, park, urban landscape resource, dendrological species.
This work aims to bring into attention the This study took place between March 2016-
necessity of preserving the natural heritage in March 2017, the focus being on the compo-
built-up areas sitional elements of Circus Park: vegetation
Green spaces contribute to the harmony of (trees, shrubs, flowering plants), the water
urban architecture and also add to the aesthetic features, walkways - tracking their evolution in
of the urban landscape, thus being an essential time through various photographs (photo
element of the human habitat (Godeanu, 2013). archive architect Valentin Donose - Photos 1,
In this regard we chose a park as the purpose of 2). The vegetation study was based on both a
this study, which through surface, composi- quality and quantity inventory. The quality
tional elements and functions is an essential inventory is based on identifying all the plant
component of the natural heritage of any city. species through field inspections during the
Circus Park (17 ha) is one of outstanding green four seasons of the year (2 field trips per
areas of Bucharest sector 2. It was created season) to phenologically highlight the vegetal
between 1960 and 1961 after the plans of species and their viability (Felicia Iliescu,
architect Valentin Donose, within the Design 1998; Doniţă N. et al., 2004; Ciocârlan, 2000).
Institute “Proiect București”. The quantity inventory was based on an
This park is a classic example (didactic even) established method: Braun-Blanquet (Van
of the idea that man is instinctively attracted by DerMaarel E., 1975) with which you can
water and trees and the appealing spaces that describe vegetal associations. Following this
these elements create and define. The studied method led to identifying some valuable
elements cover a never-ending palette of trees, compositional areas of the park in which the
shrubs, bushes, lianas which vary from natural plant material is predominantly made of trees
wild to clearly man crafted architectural shapes. and shrubs. The quantity was measured through
All of these aspects bestow the value of an surveys (4 surveys per field trip), each covering
urban landscape resource upon this park. an area of 200 m2 (the standard surface area
615
used in grouping trees and shrubs). This (from Lacul Tei Blvd. and Stefan cel Mare
method concludes the structural and Blvd.) in link with external circulation areas
quantitative associations of plants through a and with demands of neighbouring areas (ex:
combined scale of abundance - dominance after high population density of Lacul Tei and Stefan
Braun - Blanquet: + - few elements with low cel Mare quarters), linking these quarters and
coverage; 1 - many elements with low coverage ensuring circulation flow by leading to areas
or few elements with great coverage ( up to which serve different functions (children
10% of the surface); 2 - many elements with playgrounds, rest areas etc.) The sparse
coverage between 10-25%; 3 - various number vegetation which was in place before 1960 was
of elements with cover between 25-50%; 4 - kept as well. It was composed mainly from a
various number of elements with cover few elements of pyramidal poplar trees which
between 50-75%; 5 - various number of remain at the base of the hill and a white poplar
elements with cover between 75-100% (Doniță, at the edge of the park towards Lacul Tei Blvd.
Cocioabă, 2007). (this particular poplar is an outstanding element
and is protected by law to this day).
In the present day, following this study we
found 33 species of trees, 12 species of shrubs
and 32 species of plants. Among the better
represented species of trees are: Tilia
tomentosa, Platanus × acerifolia, Acer
saccharinum, Betulla verucosa, Populus
piramidalis, Quercus rubra, Salix alba, Ginkgo
biloba, Taxodium distichum, Pinus nigra etc.
(Photos 3, 4, 5).
Photo 3 - Planted areas around the lake - year 2000

2 (source: archive photo arh. V. Donose
General view of the park from Bd. Lacul Tei direction:

photo 1- year 1960 (source: arh. V. Donose);
photo 2- year 2000
The initial project of Circus Park was

outstanding through perfectly combining alley
paths and combinations with planted areas
which take the terrain architecture and the lake
presence into account. The circulation areas
respond to functional demands: granting access
Photo 4 - Planted areas around the lake - year 2016
to visitors from carefully chosen key points (original)
616
In the case of shrubs we identified 12 species, Survey 2 (200 m²): Trees and shrubs level (A/a) of
among which: Buxus sempervirens, Spiraea access area from Lacul Tei Blvd:
vanhouttei, Forsithia sp. Hibiscus sp., pictured Nr. Name A/a AD¹
crt
below (Photo 6). The analysed shrubs are 1. Quercus rubra A 60%
located near the main entrance into the park, 2. Tilia tomentosa A 25%
which would currently require some 3. Crataegus monogyna a 10%
reconditioning interventions. 4. Forsithia sp. a 5%
A particular problem of the study was ¹abundance - dominance
identifying valuable compositional areas
through exact grouping of plant species (inland We consider that the viability of a park is kept
and foreign), areas in which not only the in place by identifying, preserving and recon-
morphological characteristics of plants was ditioning, where necessary, of rich, aesthe-
taken into account, but also the rigors of tically valuable vegetation areas.
landscape architecture (Table 1). This aspect must be present in any course of
rehabilitating green areas which constitute
fundamental elements of natural heritage in any
city.
CONCLUSIONS
This study recommends that the original

concept ideas of the initial plan of the park be
maintained, as well as the variety of the
existing dendrological species.
Circus Park is an important component of the
Bucharest natural heritage. It’s value, authen-
ticity, and uniqueness deserve to be conserved.
Photo 5 - Taxodium distichum, year 2017 (original)
ACKNOWLEDGEMENTS
I would like to express my sincere gratitude to

the landscape architect Valentin Donose, the
author of the Circus Park, and to the Design
Institute “Proiect București” in which he
carried out his professional activity and which
serves as a personal archive.
Photo 6 - Buxus sempervivens - access area from Lacul REFERENCES

Tei Blvd.(source: archive photo arh. V. Donose, 2000)
Ciocârlan V., 2000. Flora ilustrată a României, Editura
In table 1 we cover two examples of important Ceres, Bucuresti.
compositional areas positioned along the alley Doniţă N., Geambașu T., Brad R., 2004. Dendrologie,
„Vasile Goldiș” University Press, Arad.
which links the two quarters (Stefan cel Mare
Doniţă N., Cocioabă S., 2007. Fitocenologie integrată şi
and Lacul Tei). Vegetaţia României, Editura Printech, Bucuresti.
Godeanu S., 2013. Ecologie aplicată, Editura Academiei
Table 1. Identifying valuable compositional areas - Romane, Bucuresti.
Circus Park (original, 2017) Iliescu, Ana-Felicia, 1998. Arboricultura ornamentala,
Survey 1 (200m²): Trees and shrubs level (A/a ) of Editura Ceres, Bucuresti.
access area from Lacul Tei Blvd: Van Der Maarel E., 1975, The Braun-Blanquet approach
Nr.cr Name A/a AD¹ in perspective, Vegetatio, 30 (3): 213-219.
t.
1. Ginkgo biloba A 60%
2. Quercus rubra A 35%
3. Crataegus monogyna a 5%
617
618
LEAF STOMATAL PARAMETERS OF IRIS GERMANICA L.

INFLUENCED BY CULTIVAR AND ARBUSCULAR MYCORRHIZAE
INOCULATION IN FIELD CONDITIONS, ROMANIA
Ioana CRIŞAN, Roxana VIDICAN, Vlad STOIAN, Sorin VÂTCĂ, Andrei STOIE

3-5 Manastur Street, 400372, Cluj-Napoca, Romania
Abstract
The aim of this study was to investigate if supplementary inoculation with arbuscular mycorrhizae of Iris germanica
plants in field conditions has influence on key stomata parameters that are known to determine the maximum leaf
diffusive conductance of CO2 to the site of assimilation as well as water use efficiency. Six Iris germanica cultivars
(‘Black Dragon’, ‘Blue Rhythm’, ‘Sultan’s Palace’, ‘Lime Fizz’, ‘Pinafore Pink’, ‘Pure As The’) were inoculated at
planting in autumn with the following arbuscular mycorrhizae fungi species: Funneliformis mosseae (Glomus
mosseae), Funneliformis geosporus (Glomus geosporum), Claroideoglomus clarodeum (Glomus claroideum),
Rhizophagus intraradices (Glomus intraradices), Glomus microaggregatum. Microscopic examination revealed that
both inoculated and non-inoculated plants presented AM root colonization after entering in vegetation. Analysis of leaf
imprints collected in spring showed that inoculation with arbuscular mycorrhizae determined a decrease in stomatal
density but increase of guard cell length. Inoculated plants presented higher potential stomatal conductance index.
Key words: physiological parameters, ornamental geophyte, AMF inoculation, root.
INTRODUCTION shown to be accompanied by increase in sto-

matal conductance (Augé et al., 1992; Augé et
Stomata are involved in regulation of water and al., 2008) or density (Chitarra et al., 2016) for
carbon dioxide balance of plants. Maximum several cultivated species. Regarding the effect
stomatal conductance to water vapor and CO2 of AMF on Iris plants, it was demonstrated the
is associated with stomata density and stomata promoting effect of Glomus mosseae on
size (Franks et al., 2017). During the evolution photosynthesis rate (Chen et al., 2014). Also,
of plants, changes in stomata density and size inoculation with Diversispora epigaea, Glomus
were linked to fluctuations in atmospheric CO2 aureum, Rhizophagus irregularis, Rhizophagus
levels (Franks et Beerling, 2009), this is why clarus influenced physiological processes of
how future elevated CO2 levels might affect Iris pseudacorus plants because it was
plants is subject of high attention (Hepworth et observed an improved phytoremediation
al., 2015). In addition, recent studies showed capacity (Wężowicz et al., 2015).
stomata parameters to be reliably linked to Previously, differences in stomatal density of
drought-resistance in some cultivated monocot Iris leaves were studied in terms of histological
plants (Li et al., 2017; Hughes et al., 2017). gradients within the leaves of the plant in
Current understanding of molecular control of different light conditions (Pazourek, 1970) or in
these leaf structures already allowed resear- order to highlight variations between Iris
chers to modify stomata density by manipu- species, subspecies or clones (Miljkovi et al.,
lating epidermal pattering factor family of 2013; Ghasemi et al., 2014; Kandemir et Çelik,
secreted signaling peptides which intervene in 2017). Thus, although previous studies were
regulation of stomata development (Hepworth able to put in evidence variations in stomatal
et al., 2015; Hughes et al., 2017). Previous parameters in Iris plants the influence of other
studies indicate that AMF inoculation could factors was little explored so far.
have influence on primary physiological Iris germanica leaves present epicuticular wax
functions of leaves as well. For example, root structures as film or platelets with regeneration
colonization with AMF species was already type following concentric-layer formation and
619
platelets (Koch et al., 2009). In the genus Iris spring 2017, conducted at the O.S.P.A. Cluj
leaf stomata observed is of anomocytic and showed a clay loam soil type with 6.72 pH, low
tetracytic type. In addition, under microscope humus level (1.35%) and good NPK supply (N
Iris leaf can present papillae on the outer 0.461%, P 68 ppm, K 312 ppm). Sum of basic
periclinal wall of epidermal cells as well as cations through Kappen method was 20.96
bulliform cells, while prism-shaped styloids of (me/100 g of soil), while hydrolytic acidity
calcium oxalate occur in idioblasts in the through extraction method was 1.92 (me/100 g
mesophyll and are arranged parallel to long soil). Basic cation saturation level determined
axis of the leaf often being visible through by calculation was 92. Granulometric analysis
epidermis. Micro-morphology leaf features using Kacinscki method was: coarse sand
hold taxonomic importance in genus Iris (Wu 14.42, fine sand 25.08, dust I 7.95, dust II
et Cutler, 1985; Mitić et Pavletić, 1995; 13.65 and clay 38.90. Low dose of NPK
Çölgeçen and Tug, 2006; Wang et al., 2010; fertilizer was applied before bloom and wood
Kandemir et Çelik, 2017). ash was applied in autumn and spring.
In all monocots stomatal development begins The bifactorial experiment established in
with an asymmetric mitosis in a meristemoid- October 2016 was organized in randomized
mother cell, which gives rise to a larger blocks with three replicates: factor A - Iris
stomatal-lineage ground cell and shorter germanica cultivars, with six levels (a1 =
meristemoid that directly forms a guard-mother ‘Black Dragon’, a2 = ‘Blue Rhythm’, a3 =
cell. Each guard-mother cell undergoes ‘Sultan’s Palace’, a4 = ‘Lime Fizz’, a5 =
symmetric mitosis to form a pair of guard cells ‘Pinafore Pink’, a6 = ‘Pure as The’) and factor
that rapidly cease expansion compared with the B – the treatment applied with two levels (b1 =
greater enlargement of pavement cells as leaves non-inoculated at planting and b2 = inoculated
grow. Because in monocots with linear leaves at planting with 13 grams mycorrhizal products
and parallel venation, stomata occur in per rhizome). The two commercial mycorrhizal
epidermal cell files that develop basipetally products special destined for use in ornamental
from an intercalary meristem (Rudall et al., plants were imported from U.K. and contained
2017), there is a histological gradient along the the following AMF species: Funneliformis
leaf blade. mosseae (Glomus mosseae), Funneliformis
The aim of the study was to investigate the geosporus (Glomus geosporum),
influence of inoculation in field conditions with Claroideoglomus clarodeum (Glomus
a consortium of five AMF species on key claroideum), Rhizophagus intraradices
stomata characteristics such as stomata density (Glomus intraradices), Glomus
and guard cell length for six Iris germanica microaggregatum. In addition to the AMF
cultivars. spores, the products contained excipients such
Results of this study could help in identifying if as organic materials, humates and auxins along
AM inoculation might optimize basic physiolo- a few others. Between the three blocks of
gical processes that could help plants cope with inoculated plants and the three blocks of non-
increasing atmospheric pollution and drought, inoculated plants was ensured 7 m, sufficient to
both known to influence plant development prevent potential cross transport of AM
within the context of current climate trend. propagules (Powell, 1979), also tools were
washed after working with each set of plants.
MATERIALS AND METHODS Microscopic examinations were conducted with
optical microscope (Figure 1c) ML-4M made
The experimental field was located in the Agro- in Romania. Images were taken with Bresser
Botanical Garden of UASVM Cluj-Napoca camera and microscope (Figures 2, 5).
Romania, elevation (AMSL) 380-430 m, with The success of root colonization was confirmed
average annual temperature 8.1°C and average in 2017 under microscope (Figure 2) using the
sum of annual precipitation 635 mm (Index staining technique of Stoian et Florian (2009)
Seminum USAMV Cluj). and colonization estimation method
The physical-chemical analysis of the soil (https://www2.dijon.inra.fr/). Using clear polish
collected when plants entered vegetation in and tape method (Palasciano et al., 2005), in
620
May 2017 were collected leaf imprints from (2007) and presented as density per square mm.
middle segment of the leaf blade surface facing Guard cell length was measured with eye piece
south (Figure 1c). reticle. Stomata were considered in the field of
For each leaf imprint sample, the number of view only if the entire ostiole was visible.
stomata was counted according to Vâtcă et al.
a b c
Figure 1. Iris germanica ‘Blue Rhythm’: a) rhizome at planting; b) plants in spring; c) leaf imprint sample
In total for all six Iris germanica cultivars, that looked rather discontinuous or patchy
stomata were counted on more than 1900 along root length.
microscopic fields of view and over 5700 The fresh, washed, and unstained thick roots of
stomata were measured for guard cell length. first order were white but thinner roots of
Leaf imprints were collected also from several higher order, were darker in color indicating
other rhizomatous cultivated species (Iris root colonization causing a darkening of the
pallida, Iris pseudacorus, Iris sibirica and Iris cortical cells under the pigment released by
chrysographes) grown in Agro-Botanical collapsed arbuscules (Fester et al., 2002). All
Garden UASVM Cluj-Napoca, for comparison AMF structures were present: extraradical
purposes and analyzed similarly. hyphae, hyphopodium and inner root coils at
Based on the microscopic examinations were the entrance points, young and collapsed
calculated: arbuscules, vesicles (oblong and round) and
1. PCI = L2 × SD × 10-4 (Holland et spores as well as a few loose sporocarps.
Richardson, 2009), where PCI = potential Under microscope was identified Arum type
conductance index, L = guard cell length proliferation as well as intermediate Arum-
(μm), SD = stomata density per 1 mm2; Paris type proliferation. Spreading patterns
2. a% = [(100 × mA3) + (50 × mA2) + (10 × varied slightly among cultivars. Thus, it was
mA1)]/100 (Trouvelot et al., 1986), where noticed in some segments a preponderant linear
a% = arbuscule abundance in mycorrhizal spreading within the roots of ‘Pure As The’,
parts of the root fragments, based on arbuscules preferentially along sieve elements
percentages of m rated according to in ‘Lime Fizz’ or in the outer root layers in
methodology. ‘Sultan’s Palace’, abundant hyphae coils within
Data was analyzed with Microsoft Excel, roots of some ‘Black Dragon’ plants.
Mycocalc and Origin. Arbuscules abundance in mycorrhizal parts of
root fragments of the six Iris germanica
RESULTS AND DISCUSSIONS cultivars was slightly increased from average
a% = 44.09 in non-inoculated plants to a% =
In 2017 at the end of active vegetative growth 44.45 in inoculated plants, hinting to a higher
of plants (end of spring) both inoculated and transfer occurring between plants and
non-inoculated Iris germanica plants presented arbuscular mycorrhizae fungi following
root colonization confirmed under microscope inoculation treatment.
621
a
b
Figure 2. Microscopic view: a) colonized Iris germanica roots, some with pigmentation
due to apocarotenoids released by collapsed arbuscules; b) roots after staining
In five out of six Iris germanica cultivars, with about 10 stomata/mm2, from an average
stomata density decreased as a result of density of 63.83/mm2 in non-inoculated plants
inoculation with arbuscular mycorrhizae to 53.12/mm2 in inoculated plants. Highest
(Figure 3). The only cultivar that presented a decrease of stomata density due to inoculation
slight increase in stomata density due to was observed in ‘Sultan’s Palace’ followed by
inoculation was ‘Black Dragon’ from ‘Pinafore Pink’ and ‘Pure As The’. The
61.41/mm2 in non-inoculated plants to smallest decrease in stomata density caused by
61.95/mm2 in inoculated plants. On average, inoculation was found in ‘Lime Fizz’ followed
stomata density decreased in inoculated plants by ‘Blue Rhythm’.
Figure 3. Stomata density (per mm2) and guard cell length (µm) in six Iris germanica cultivars:
1) ‘Black Dragon’; 2) ‘Blue Rhythm’; 3) ‘Sultan’s Palace’; 4) ‘Lime Fizz’; 5) ‘Pinafore Pink’; 6) ‘Pure as The’
non-inoculated (Myco-) and inoculated with arbuscular mycorrhizae (Myco+)
In all inoculated Iris germanica cultivars guard Potential stomatal conductance index increased
cell length increased (Figure 3) on average with due to inoculation in all studied Iris germanica
3.56 µm. Non-inoculated plants on average had cultivars (Figure 4), from the average of 2.93 in
a stomata length of 21.39 µm while inoculated non-inoculated plants to 3.31 in the inoculated
cultivars had an average stomata length of plants. The highest increase was observed in
24.95 µm. The largest increase in guard cell ‘Pinafore Pink’ (from 2.30 to 3.06). This
length linked to inoculation was observed in cultivar also experienced one of the highest
‘Pinafore Pink’ that had the second highest decrease in stomata density and highest
decrease of stomata density as a result of increase in guard cell length due to inoculation.
inoculation followed by ‘Sultan’s Palace’. The Also, it can be noted that non-inoculated plants
slightest increase in guard cell length due to of ‘Pinafore Pink’ presented the smallest guard
inoculation was observed in ‘Black Dragon’ of cell length among all cultivars studied. Two
only 1.6 µm on average. cultivars presented very similar increase of
potential stomata conductance due to
622
inoculation: ‘Black Dragon’ (from 3.10 to 3.59) cultivars, with an increase of PCI due to
and ‘Blue Rhythm’ (from 2.91 to 3.35). Both inoculation from 3.06 observed in non-
these cultivars had either a slight decrease in inoculated plants to 3.40 observed in inoculated
stomata density or a very small increase in plants (Figure 4).
stomata density due to inoculation. The least Although stomata density did not increase, on
increase in potential stomata conductance index contrary decreased in inoculated plants with
was observed in ‘Pure As The’, from 2.54 to one exception, the potential stomatal conduc-
2.61, followed by ‘Sultan’s Palace’ with an tance increased in all studied Iris germanica
increase from 3.69 to 3.85. The cultivar ‘Lime cultivars due to inoculation, because of the
Fizz’ with yellow flowers, situated between the increase in guard cell length.
dark-flowered cultivars and the light-flowered
Figure 4. Potential stomatal conductance index (PCI) in six Iris germanica cultivars
non-inoculated (Myco -) and inoculated (Myco +) with arbuscular mycorrhizae
During the vegetative months prior to flowe- citing previous researches mention that plants
ring and before collecting the leaf imprint with low stomata density presented enhanced
samples there was a warm spring with water use efficiency and reduced transpiration
temperatures exceeding 20°C on no less than levels that allowed plants to grow larger
17 days during the interval 15 March - 15 May especially under water restriction conditions.
2017. The sum of precipitations was about 45 But reduced stomata could also mean lower
mm during same time interval, while the wind transpiration. Yet, transpiration is driving the
had intensities between 3-37 km/h mass flow (Hepworth et al., 2015), fact that
(https://www.wunderground.com/history). would suggest that if stomata density decreases
Plants were not irrigated and supported a under a certain level it would be expected to be
natural water regime. Agro-Botanical Garden made at the expense of nutrient accumulation.
of UASVM is situated on the steep side of The increased guard cell length ensured the
Someşul Mic river valley within Cluj-Napoca increase of potential stomata conductance index
and experiences windy conditions in spring and despite of a reduction in stomata density. Plants
fall, fact known to increase the transpiration can rely on mycorrhizae mycelia network for
and water loss in plants. increase uptake of both nutrients and water, but
In the given conditions, it can be considered mycorrhizal fungi take in exchange part of the
that plants had to find ways to cope with these carbon fixed by the plant. Thus, the inoculated
environmental challenges by optimizing their plant needs to be able to conduct photosyn-
physiological processes. Hepworth et al. (2015) thesis at optimum levels, and this is perhaps
623
why the reduction in stomata density was species were observed under microscope on
accompanied by increase in stomata length. collected leaf imprint samples (Figure 5).
Since both sets of plants (inoculated and non- Comparing the stomata density and guard cell
inoculated presented root colonization), the length of non-inoculated Iris germanica plants
observed changes might be linked to inoculated (Figure 3) with those of Iris pallida (Table 1),
species rather than those already present in the it can be observed that values for Iris
soil; also, some excipients could have played a germanica are higher. It is known that current
beneficial role as well, either directly on plant Iris germanica cultivars are tetraploids (Norris,
rooting and nutrition or indirectly by 2012) and this cultivated species is actually a
stimulating the establishment of symbiose. natural hybrid with diploid Iris pallida as one
Micromorphology differences between studied of its ancestors (Lim, 2016).
Iris germanica cultivars and other rhizomatous
c d e
Figure 5. Leaf micromorphology on imprints from cultivated rhizomatous Iris species:
a) Iris germanica; b) Iris pallida; c) Iris pseudacorus; d) Iris sibirica; e) Iris chrysographes
Ploidy level influences stomatal characteristics, All three species from subgenus Limniris (Iris
since guard cell size has been used to predict pseudacorus, Iris sibirica and Iris
the haploid level of primitive angiosperms and chrysographes) presented higher stomata
the extent of polyploidy in the present-day density than the two species from subgenus Iris
angiosperms (Willmer et Fricker, 1996). On (Iris germanica and Iris pallida).
average both inoculated and non-inoculated Iris germanica and Iris pallida are native to
plants of Iris germanica presented longer guard milder Mediterranean climate, with warm
cell length, and polyploidy of Iris germanica summer (Lim, 2016), and their stomata density
could explain this characteristic compared to and guard cell length have similar values. The
diploid Iris pallida. However, the values are lower stomata density compared to the other
too close for a clear indication of ploidy level three species studied (Table 1) can be an
in Iris germanica based on stomata parameters indication of their known drought tolerance and
compared to Iris pallida, even more since their known preferences for dryer substrate
inoculated Iris germanica plants had slightly (Beresford-Kroeger, 2004). On Iris germanica
smaller stomata density than Iris pallida. leaves were observed large papillae (no more
Ghasemi et al. (2014) trying to find if stomata than one per epidermal cell) while stomata over
parameters can be used in several Iris taxa from the leaf veins are present but disposed sparsely.
Iran to predict ploidy level, reached same Iris sibirica and Iris chrysographes originate
conclusion that, environmental factors have a from colder regions of Eurasia and respectively
strong influence on stomata characteristics that Asia (White et al., 1997). Under microscope
would make this approach less exact in these two present smaller epidermal cells
predicting ploidy level in Iris. compared to other Iris plants from this study
and abundant leaf papillae. Both present
624
smaller guard cell length and high stomata species is used for water gardens and grows
density. It is known that smaller stomata occur best in marshes (White et al., 1997), fact that
at higher frequency, fact that determines the can be linked to observed stomata parameters
total possible pore area for leaf to be similar that indicates to higher water requirements
(Willmer et Fricker, 1996). Iris pseudacorus known that this species has (Jacobs et al.,
presents both a large number of stomata per 2011).
leaf unit area as well as longer guard cells. This
Table 1. Stomata parameters in four Iris species from Agro-Botanical Garden UASVM Cluj-Napoca
Species SD (mm2) L (µm) PCI
Iris pallida Lam. 56.33 20.83 2.44
Iris pseudacorus L. 82.11 23.77 4.64
Iris sibirica L. 97.59 17.28 2.91
Iris chrysographes Dykes 72.34 16.13 1.88
SD – stomata density, L – guard cell length, PCI – potential stomatal conductance index
Summarizing the observations and findings of germanica cultivars either inoculated or non-
this research, firstly can be suggested that inoculated. This comes to show that in field
patchy colonization pattern observed in both conditions colonizing patterns of plants can be
sets of Iris germanica plants (inoculated and different.
non-inoculated) could be due to soil texture, The yellowish tint of different intensities
particularly abundant clay component, that also identified in colonized Iris germanica roots of
might have caused poorer root ramification too. higher order appears in response to AM fungi
Previous studies showed that soil texture and various carotenoid degradation products
particularly clay rich soil along lime (apocarotenoids) released during collapse of
application are some of the most important arbuscules. These accumulate as hydrophobic
factors influencing AMF root colonization in droplets in root cortical cells and were
maize, sorghum and peanuts (Carrenho et al., identified in many Liliopsida species analyzed.
2007). Also, patchiness characterizes the Their function is little known but might have
distribution of mycorrhizal structures of importance in the arbuscule development and
members from genera Ambispora, function (Fester et al., 2002). This observation
Archaeospora, Diversispora, Entrophospora, indicated that Iris germanica plants were
Intraspora and Paraglomus, whereas members colonized during spring. Root samples were
of the families Gigasporaceae, Glomeraceae collected shortly after the flowering season.
and Pacisporaceae usually present a Between inoculated and non-inoculated plants
continuous mycorrhizal distribution along the was identified only a small difference regarding
root (Błaszkowski, 2012). This might as well arbuscules abundance in mycorrhizal parts of
explain the patchy colonization observed. root fragments.
The different colonization patterns observed in A direction for future investigation would be to
some cultivars can be attributed both to some investigate the seasonal variation of root
differences in root architecture between colonization in order to establish the link
cultivars as well as to possible plant genotype- between plant phenology and mycorrhiza life
AMF colonizing specificity but could not be cycle.
associated to inoculation treatment. In five out of six studied Iris germanica
Previously in Iris pseudacorus plants grown in cultivars, a decrease in stomata density was
pots on sterile substratum and inoculated with accompanied by increase in stomata length.
Diversispora epigaea, Glomus aureum, These findings are in accordance with previous
Rhizophagus irregularis and Rhizophagus patterns observed in Iris stomata. In two taxa
clarus obtained from trap cultures was reported from Turkey, Iris masia subsp. dumaniana Iris
typical Arum type spreading (Wężowicz et al., masia subsp. masica higher number of stomata
2012), but in this study conducted in field was linked to smaller length of stomata
conditions both Arum and intermediate Arum- (Kandemir et Çelik, 2017). Similarly, Ohsumi
Paris type proliferation was identified in Iris et al., 2007 found a negative correlation
625
between stomata density and stomata length in up until a certain growing factor (like
Oryza sativa plants, another monocot. inoculation) can be used to enhance plant
All Iris germanica cultivars inoculated with response and its effect cannot guarantee an
arbuscular mycorrhizae presented increased equal response among plants.
potential stomatal conductance index (Figure Expression of genes homologous to those
4). Previous studies showed that Glomus involved in the regulation of stomatal
mosseae inoculation had a stimulating effect on development in Arabidopsis known as
photosynthesis of Iris plants (Chen et al., STOMAGEN and genes encoding two
2014). Also, leaves of AMF inoculated Vigna intercellular signaling factors that act as
unguiculata had higher stomatal conductance negative regulators for stomatal development
than those of non-mycorrhizal plants before antagonistic to the first one was investigated in
and after lowering soil water potential (Augé et developing leaves of AMF inoculated tomato
al., 1992). plants.
In contrast with the results of this study on Iris It was showed that genes EPF1 and EPF2 were
plants, in controlled conditions Rhizophagus significantly modulated only in the presence of
intraradices-inoculated tomato plants presented AM symbiosis, while LeEPFL9 transcript
significantly increased number of stomata in levels were correlated with changes in stomata
mature leaves; stomatal density was almost density of Rhizophagus intraradices-inoculated
twice that of control tomato plants or plants (Chitarra et al., 2016).
Funneliformis mosseae-inoculated plants According to the results of this study,
(Chitarra et al., 2016). By comparison, in this supplementary inoculation has influence on two
study the inoculated Iris plants grown in field key stomata parameters.
conditions experienced a reduction in stomata In inoculated plants, on average stomatal
density, maybe as a strategy to reduce water density decreased but the length of guard cells
loss. Previous experiments showed that a route increased, possibly hinting to a tendency to
towards improving drought tolerance and water balance water use efficiency and increased
use efficiency without significantly affecting assimilation rate that ultimately lead to
photosynthetic capacity or nutrient potential stomatal conductance index to
accumulation by mass flow is a slight reduction increase in all inoculated cultivars.
in stomata density (Hepworth et al., 2015).
In conclusion can be said that environmental CONCLUSIONS
factors influence stomata parameters, but plant
genotype also has a strong influence as well. On average, stomata density decreased in
This can be exemplified by two observed arbuscular mycorrhizae-inoculated plants with
phenomena from this study. about 10 stomata/mm2, at the same time guard
First, different Iris germanica cultivars did not cell length increased on average with 3.56 µm.
responded identical to inoculation with The changes in stomata parameters following
arbuscular mycorrhizae, although in all cases inoculation lead to an increase of potential
there was an increased potential stomatal stomatal conductance observed in all studied
conductance index. Secondly, different cultivars.
rhizomatous species grown in Agro-Botanical Both inoculated and non-inoculated plants
Garden of Cluj retain their ancestral presented root colonization, with some close
characteristics from adaptation to their habitat values for arbuscule abundance in mycorrhizal
of origin. This is most simply explained by the parts of root fragments between the two sets of
known fact that each species maintains specific plants.
growing requirements similar with the More studies should be conducted to assess the
environment they evolved in, just as stomata physiological response of plants to mycorrhiza
density in different Iris species mentioned inoculation in field conditions, in order to
above can be easily linked to the way their further define the practicality of its application
physiological processes were adapted to for ornamental irises as well as irises destined
ecological niche they preferentially occupied. to obtaining orris oil.
This suggests there might be a certain threshold
626
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628
PRELIMINARY TESTING OF SRAP PRIMERS IN FOUR

RANUNCULACEAE SPECIES FROM ROMANIA
Cristina D. KELEMEN1, Orsolya BORSAI1, Katalin SZABO2, Doina CLAPA2,
Monica HÂRŢA2, Doru PAMFIL1,2
1
University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca, Department of
Horticulture, 3-5 Mănăştur Street, Cluj-Napoca, Romania
2
University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca, Life Sciences Institute,
Research Centre for Agricultural Biotechnology affiliated at Romanian Academy,
3-5 Mănăştur Street, Cluj-Napoca, Romania
Abstract
The main aim of this paper was to identify the most reliable primer pairs out of the 64 used in order to investigate the
genetic diversity of four medicinal Ranunculaceae species as follow: Aconitum moldavicum, Anemone transsilvanica,
Ranunculus carpaticus and R. platanifolius. Amplified products were separated on 1.6% agarose gel and a total
number of 886 DNA fragments were visualized by EtBr staining. After primer combinations screening, 27 primer pairs
were selected based on clarity, reproducibility of the amplified bands and high rates of polymorphism for further
research. The selected SRAP primer pairs resulted in 553 informative fragments with an average number of 20.5 per
primer combination. The most efficient primer combinations were Me1/Em1, Me3/Em3, Me7/Em4, Me7/Em5,
Me7/Em8, Me8/Em1 and Me8/Em5 which generated a total number of 201 visible DNA fragments. The current
research represents a preliminary study for further conservation perspectives regarding the genetic diversity of several
medicinal plants belonging to the Ranunculaceae family.
Keywords: Ranunculaceae, genetic diversity, primer testing, SRAP.
INTRODUCTION In Romania, the Ranunculaceae family is

represented by 23 genera and approximately
Ranunculaceae Juss. (buttercup family) is one 110 species including endemic plants such as
of the early lineages in flowering plants Anemone transsilvanica Fuss (Săvulescu,
distributed worldwide, most commonly in the 2007). Many local plants belonging to
temperate and cold areas of the northern Ranunculaceae family (Aconitum moldavicum
hemisphere (Heywood et al., 2007). The family Hacq., Anemone transsilvanica, Ranunculus
including 56 genera and approximately 2,500 carpaticus Herbich, R. platanifolius L.) have
species used as medicinals, spices, and been used as human and veterinary medicine to
ornamentals, however, most of them are highly treat various ailments such as bronchitis,
poisonous (Nyirimigabo et al., 2014; Aslam et cough, diarrhea, fever, hepatitis, gout,
al., 2012). The most representative genera of rheumatism, and skin diseases (Tămaș, 2005).
the buttercup family are Aconitum, Anemone, However, little is known concerning their
Clematis, Delphinium, Ranunculus, and genetical diversity and conservation.
Thalictrum which account approximately 90% The main purpose of this work is to assess the
of the species (Heywood et al., 2007). From genetic diversity at molecular level of A.
genetic point of view this family has a great moldavicum, A. transsilvanica, R. carpaticus
chromosomal variation in numbers, sizes and and R. platanifolius using sequence-related
structures, including species with 2n=14 (e.g. amplified polymorphism (SRAP) markers in
Hepatica asiatica Nakai, Ranunculu sceleratus order to establish some conservation strategies
L.), 2n=16 (e.g. Ranunculus cantoniensis DC), for these species. SRAP primers are
2n=18 (e.g. Clematis brachyura Maxim), and considering to be more suitable to revealing
2n=42 (e.g. Thalictrum aquilegiifolium var. genetic diversity among related species than
sibiricum Regel & Tiling) (Chung et al., 2013). amplified fragment length polymorphism
629
(AFLP), simple sequence repeats (SSR), inter- (2015). Concentration and purity of the
simple sequence repeat (ISSR) or even random extracted DNA were determined using
amplified polymorphic DNA (RAPD) markers NanoDrop1000 spectrophotometer. The SRAP
(Budak et al., 2004). Therefore, preliminary analysis was carried out according to
testing of SRAP primers was necessary to be previously established protocols by Li and
able to select the appropriate primer Quiros (2001). In order to select the most
combinations on a large-scale use to reliable primer combinations for further
characterize the genetic diversity of the studied analyses, 64 different primer combinations
species. Although, previous scientific reports were employed in this assay using eight
show intra- and interpopulation genetic forward and eight reverse primers (Table 2).
diversity of various Ranunculaceae species Table 2. SRAP primers sequence
[Ranunculus acris (Odat et al., 2004), R.
Forward primers
cabrerensis (Cires et al., 2013), R. kuepferi Me1 F: TGA GTC CAA ACC GGA TA
(Cosendai et al., 2013) and R. reptans (Fischer Me2 F: TGA GTC CAA ACC GGA GC
Me3 F: TGA GTC CAA ACC GGA AT
et al., 2000; Prati et al., 2016)] employing Me4 F: TGA GTC CAA ACC GGA CC
AFLP or RAPD primers, this is the first report Me5 F: TGA GTC CAA ACC GGA AG
Me6 F: TGA GTC CAA ACC GGA CA
regarding to assessment of genetic variation in Me7 F: TGA GTC CAA ACC GGA CG
this family using SRAP markers. Me8 F: TGA GTC CAA ACC GGA CT
Reverse primers
Em1 R: GAC TGC GTA CGA ATT AAT
MATERIALS AND METHODS Em2 R: GAC TGC GTA CGA ATT TGC
Em3 R: GAC TGC GTA CGA ATT GAC
Em4 R: GAC TGC GTA CGA ATT TGA
Plant samples were collected from their natural Em5 R: GAC TGC GTA CGA ATT AAC
Em6 R: GAC TGC GTA CGA ATT GCA
habitats in two districts from Romania (Brașov Em7 R: GAC TGC GTA CGA ATT CAA
and Hunedoara) namely at Mt. Postăvaru Em8 R: GAC TGC GTA CGA ATT CAC
(45°35.215´ N, 25°433.146´ E) and Mt. Stâmba
(46°12.508´ N, 22°51.354´ E). The voucher The polymerase chain reaction was performed
specimens have been stored in the Herbarium in 15 μL reaction mixture containing 1 × Green
collection at the University of Agricultural Buffer, 1.5 mM MgCl2, 200 µM of dNTPs, 0.3
Sciences and Veterinary Medicine of Cluj- µM of both forward and reverse primers
Napoca, Romania. The location and the (Generi Biotech), 1 U of Taq DNA polymerase
voucher specimen numbers of the species are (Promega) and approximately 50 ng of
shown in Table 1. template DNA. Amplifications were made in a
Table 1. Voucher specimen numbers of the
Corbette Research PalmCycler with an initial
Ranunculaceae species and their geographical origins step at 94°C for 5 min and five cycles of 1 min.
at 94°C, 1 min. at 35°C, and 1 min. at 75°C.
Species name VSNb) Longitude E Latitude N
Aconitum The following 35 cycles consisted of 94°C for
CLA30049
moldavicum Hacq.
22°51.354´ 46°12.508´ 1 min., 50°C for 1 min., and 72°C for 1 min.,
Anemone CLA30047 25°433.146 45°35.215´ with a final extension at 72°C for 10 min. The
transsilvanica FUSS.
Ranunculus
PCR reactions were repeated to ensure the
CLA30044
carpaticus Herbich.
25°433.146 45°35.215´ reproducibility of the DNA. Amplified
Ranunculus
CLA30040 25°433.146 45°35.215´
products were separated on 1.6 % (w/v)
platanifolius L.
b)
agarose gels in 1.0 × TAE buffer at 115 V, for
Note: Voucher specimen number
approximately 2 hours. A 100-bp DNA ladder
was used as molecular weight marker in order
The harvesting leaves were air dried at room
to confirm the appropriate SRAP markers. The
temperature and stored at -20°C until
electrophoretic profiles were stained with
processing. To find out the most suitable SRAP
1µg/mL EtBr form 20 to 30 minutes and
primer combinations four randomly selected
photographed with Biospectrum AC
samples were used one of each species.
The obtained images were processed with Total
In order to extract the genomic DNA, its
Lab 120 software (Figure 1).
isolation was made by applying the CTAB
.
method as described by Lodhi et al. (1994) and
improved by Pop et al. (2003) and Szabo et al.
630
Figure 1. Sequence-related amplified polymorphism (SRAP) profiles obtained
from 16 primer combinations of four species
.
RESULTS AND DISCUSSIONS and reproducibility of the amplified bands
together with the rate of polymorphism.
The screening of 64 SRAP primer pairs (eight Therefore, after the screening of the amplified
forward and eight reverse) generated a total products, 37 primer pairs were discarded out of
number of 886 DNA fragments ranging from 1 the 64 tested, because they either yielded no
to 35 bands per primer combination. The amplification or no polymorphic patterns.
average number band per primer set recorded The remaining 27 primer combinations
was 13.9. The size of amplification products produced a total number of 553 DNA
ranged from 400-3,200 bp. fragments with an average number of 20.5 clear
In order to select the most efficient primer bands per primer pairs (Table 3).
combinations for subsequent analyses, the main
criteria taken into consideration were the clarity
Table 3. The remaining 27 primer combinations
Primer Number of amplified fragments Total Average
A. A. R. R. number of number of
combination bands bands
moldavicum transsilvanica carpaticus platanifolius
Me1/Em1 7 6 7 5 25 6.3
Me1/ Em3 3 6 5 3 17 4.3
Me1/ Em4 5 6 6 4 21 5.3
Me1/Em6 4 4 6 1 15 3.8
Me1/ Em7 5 6 6 1 18 4.5
Me2/Em2 3 4 3 6 16 4.0
Me2/ Em3 6 1 4 3 14 3.5
Me3/ Em2 6 2 1 2 11 2.8
Me3/Em3 7 6 10 4 27 6.8
Me3/ Em4 6 6 1 1 14 3.5
Me3/ Em5 3 8 4 4 19 4.8
Me4/ Em1 2 5 8 3 18 4.5
Me4/Em4 0 2 6 5 13 3.3
Me4/ Em6 3 4 7 4 18 4.5
Me4/ Em7 2 5 7 7 21 5.3
Me5/Em5 6 6 6 4 22 5.5
Me6/ Em4 3 6 7 3 19 4.8
631
Number of amplified fragments Total Average
Primer
A. A. R. R. number of number of
combination
moldavicum transsilvanica carpaticus platanifolius bands bands
Me6/ Em5 4 8 6 5 23 5.8
Me6/Em6 7 3 4 5 19 4.8
Me7/ Em4 7 6 6 9 28 7.0
Me7/ Em5 6 4 8 7 25 6.3
Me7/ Em6 5 3 5 8 21 5.3
Me7/ Em8 10 7 6 8 31 7.8
Me8/ Em1 9 4 8 9 30 7.5
Me8/ Em6 3 2 5 10 20 5.0
Me8/ Em5 7 10 10 8 35 8.8
Me8/Em8 6 1 6 0 13 3.3
Total number
135 131 158 129 553
of bands
Average number
10.4 10.1 8.8 9.9 20.5
of bands
The most representative SRAP profiles were generated a total number of 201 visible DNA
obtained with the primer combinations fragments with a mean value of. 28.7 clear
Me1/Em1, Me3/Em3, Me7/Em4, Me7/Em5, bands per primer set (Figure 2).
Me7/Em8, Me8/Em1 and Me8/Em5 which
Figure 2. Electrophoretic profiles of four species obtained with eight primer combinations
A. moldavicum samples show good results with platanifolius were Me7/Em4, Me7/Em6,
the primer combinations: Me7/Em8, Me8/Em1, Me7/Em8, Me8/Em1, Me8/Em6.
Me8/Em5, Me7/Em4, Me6/Em6, while for A. Regarding the number of amplified bands per
transsilvanica the most effective primers pairs primer pair, 13 primer combinations were
were founded to be: Me3/Em5, Me6/Em5, selected as the most promising for A.
Me7/Em8, Me8/Em5. For R. carpaticus the moldavicum, A. transsilvanica and R.
following combinations were the most platanifolius, respectively, while for R.
informative: Me3/Em3, Me4/Em1, Me7/Em5, carpaticus 18 combinations were the most
Me8/Em1, Me8/Em5. Besides, the most representative (Figure 3).
effective primer combinations for R.
632
Figure 3. The representative combinations of A. moldavicum, A. transsilvanica, R. carpaticus and R. platanifolius
Similar studies were carried out on Carthamus A. moldavicum, A. transsilvanica, R. carpaticus

species as reported by Mokhtari et al., 2013, and R. platanifolius also for in situ and ex situ
who found that the most informative SRAP conservation perspective. Furthermore, in this
primer combinations were Me4/Em1 and study have been shown that the selected SRAP
Me5/Em2. markers combinations represent a powerful tool
Another report published on Citrus species and highly contribute to perform future analyze
show, that the most effective primers pairs that intra/interspecies and intra/interpopulation
worked, were Me3/Em2 and Me6/Em4 (Uzun et genetic diversity.
al, 2009). To the best of our knowledge, SRAP markers
Moreover, based on the number of polymorphic were used for the first time to analyze genetic
fragments the most representative SRAP primer diversity of species belonging to the
combinations reported for 76 Vitis species were Ranunculaceae family.
Me1/Em7, Me3/Em2, Me5/Em6 (Guo et al.,
2012). Likewise, Huang et al, 2017 reported ACKNOWLEDGEMENTS
that for Stylosanthes species the best working
SRAP primer pairs for genetic variation This research work was partly supported by the
analysis were Me1/Em2, Me6/Em7, Me8/Em1 PN-II-PT-PCCA2013-2014, project no.
and Me9/Em2 (Huang et al., 2017). 168/2014 GERMPLUM.
Therefore, it can be concluded that the
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POSSIBILITIES FOR SMALL-SCALE COMPOSTING

OF HORTICULTURAL PLANT WASTES
Milena YORDANOVA, Erik MINCHEV, Radoslav BORISOV
University of Forestry, Faculty of Agronomy, 10 Kliment Ohridski Blvd,

1797, Sofia, Bulgaria,
Abstract
The aim of the study was to investigate the possibilities for composting in a small suburban farm, where also vineyards,
orchards and vegetable crops are grown. The experiment was carried out in the period 2016-2017 in the experimental
field on University of Forestry - Sofia. In the spring of 2016 were selected plant wastes from viticulture and horticulture
and were built two compost piles – one only with plant residues (grape vine canes, fruit twigs and grass windrow) and
one with rabbit manure,(grape vine canes, fruit twigs and grass windrow+ rabbit manure). In the autumn of 2016 was
built a compost pile only with plant residues from vegetable field (frostbitten tomato and pepper stems and fresh leek
residues). During composting period it was monitored the temperature regime in the compost piles. They achieved high
temperature (60 and above 60 °C). The active phase of composting, in the three compost piles lasted for about a month.
At the beginning and after the 6 months period were measured pH and C/N ratio in the compost piles and changes were
established. After the 6 months period the C/N ratio was below 20/1, which is an indicator of mature compost. The pH
in the spring piles dropped to 7.5-7.6 while the autumn pile remained alkaline - 8.4.
Key words: composting, grape vine canes, fruit twigs, rabbit manure, vegetable residues.
INTRODUCTION has been beneficial (Maniadakis K. et al.,

2004).
Composting is a natural process of decom- In aerobic composting, one of the most used
position of organic waste using microorganisms methods is the ordering layers by layers of the
and under strictly controlled conditions, at the various materials for the construction of the
end of the process to give a stable humus-like compost piles, alternating layers of carbon and
organic product called compost. (Misra R.V. et layers of nitrogen material, the different authors
al., 2003). indicating a different thickness of the layers.
Organic waste from agriculture - from pruning, (Misra R.V. et al., 2003; Inckel M. et al., 2005).
harvesting, manure, etc. can be transformed Aerobic composting is a dynamic process.
into material that is used to improve soil Several main factors are favoring composting
structure and provide nutrients. (Misra R.V. et processes: temperature, pH, oxygen level and
al., 2003; Román P. et al., 2015) C/N ratios.
Most of these materials can easily be harvested These factors are a key point in building
on each farm. In the presence of animals, it is sustainable practices for organic waste
well that plant waste materials to be composted management (Maheshwari D.K. et al., 2014).
are not those used as fodder (Inckel M. et al., Studies on the carbon, nitrogen, pH content are
2005; Schuchardt F., 2005). key to tracking composting processes (López-
When testing various combinations of plant González et al., 2013).
residues from tomatoes, cucumbers, eggplants When testing a different initial ratio C/N of
and pepper (composted alone or with olive tree 20/1, 30/1 and 40/1, compost with an initial
branches and vine rods), it has been found that C/N ratio of 30/1 has developed the highest
the compost piles made by mixing vegetable maximum temperature (63.5°C) (Azim K. et
crops with twigs or grapes rods have developed al., 2014).
high temperatures and the composting process To determine if the compost is mature, ie.
whether it is ready for use, different methods
637
are available. Changes in temperature, pH, C/N (large waste from last year's compost of plant
ratio, microorganisms are observed. The C/N waste and a thin layer of soil).
ratio gradually decreases and when it reaches The first compost uses a combination of brown
less than 20, the temperature also decreases materials in a ratio of 2.4/1 (grape vine
(Karadag D. et al., 2013). canes/fruit twigs). To them was added a grass
Many authors point out that the C/N ratio can windrow as a green material. The weight ratio
be taken as an indicator of maturation of of brown/green materials was 1.6/1
compost (Goyal S. et al., 2005). Other authors (brown/green).
have the opposite view because of the variety The second one also uses a combination of the
of materials that can be composted - with their above-mentioned brown materials at a ratio of
structure and accessibility. 2.2/1 (grape vine canes/fruit twigs). A
Others point out that the ratio is not important, combination of green materials was added to
but rather how much it decreases during the them - a grass windrow and a rabbit manure in
composting process. Ratio C/N=12 is indicated a ratio of 1.1/1. The total weight ratio between
for the ideal but also a C/N=15-20 ratio is also brown and green materials is 1.8/1.
acceptable (Estévez-Schvarz et al., 2012). The compost bundle in autumn is dominated by
The aim of the study was to investigate the green materials in order to develop a higher
possibilities for composting in a small suburban temperature in the compost bundle under
farm, where also vineyards, orchards and conditions of low air temperatures.
vegetable crops are grown. During composting period it was monitored the
temperature regime in the compost piles. With
MATERIALS AND METHODS compost thermometers in the middle of the
compost pile. The temperature was measured at
The experiment was carried out in the period noon hours.
2016-2017 in the experimental field on Laboratory analyzes of the content of C, N, K,
University of Forestry - Sofia. P, Mg, in starting materialsand in mature
In the spring of 2016 were selected plant compost to track changes in compost and to
wastes from viticulture and horticulture and establish its quality.
rabbit manure. In the autumn of 2016 were Calculate the ratio of C/N in the starting
selected plant residues from vegetable field. materials and in mature compost.
Three compost piles were built from the The data have been statistically processed and
crushed plant materials - two in the spring and interpreted.
one in the autumn:
V1 - a combination of grape vine canes, RESULTS AND DISCUSSIONS
fruit twigs and grass windrow;
V2 - a combination of grape vine canes, The selection of waste materials is based on
fruit twigs and grass windrow + rabbit two criteria: to be available in sufficient
manure; quantity and not to be used in animal
V3 - a combination of frostbitten husbandry (such as bedding or fodder). Manure
tomato and pepper stems and fresh leek samples were taken for analysis from all groups
residues. of animals that are kept.
It was made a pre-preparation of the starting The total weight of materials used to build the
materials - they were shredded into smaller part first compost pile was 59,500 kg and for the
by garden shredder. Shredded material contai- second compost pile was 56,050 kg.
ned finely divided particles and larger ones. The total weight of the materials used to build
Larger parts were also inserted into the the third compost pile was 218 kg.
compost piles to aid natural aeration inside the Together with the preparation of the
pile. composting materials were taken samples to
To individual compost piles were incorporated analyze the content of chemical elements.
starters - materials, making it easier to start the (Tables 1 and 2).
microbiological processes in the compost piles
638
Table 1. Available plant waste and their chemical respectively. Rainfall was also reported in July,
characteristics but they were not heavy (Figure 1).
Chemical elements
Type of Р K Mg
Season C N 60
material mg/ mg/ mg/ 49,4
% %
kg kg kg 50
Grape
vine Sping 26.56 0.753 1.02 4631 1176 40
canes 27,5
30
Fruit
Sping 25.09 0.670 0.89 2856 725
l/m2
twigs 20
Corn
Autumn 26.93 0.973 52.92 5619 1057
leaves 10
*Grass Sping/
26.01 3.355 3.62 30622 2807 0
windrow Autumn
Tomato
stems Autumn 24.11 2.225 301.28 3671 5150
frostbitten
Pepper
stems Autumn 25.15 2.454 127.60 5796 7322
frostbitten
Fresh Figure 1. Amount of rainfall in l/m2 at ten-day periods
leek Autumn 18.46 1.737 138.43 27490 3201
residues Wet periods also led to a decrease in the
* Note: grass windrow can be used for composting and for fodder.
average daily temperature at the end of May
The C/N ratio was highest in the waste material and in the first ten days of June (Figure 2). The
from fruit (37.45) and grape vine canes (35.27), precipitations recorded in the second ten days
as they are carbon materials and the grass did not lead to a decrease in the air temperature
windrow had the lowest ratio (7.75), since it (Figure 2).
was high in total nitrogen. The remaining three
macroelements were also the highest in grass 25
24 22,87 23,02
windrow (P 3.62 mg/kg; K 30622 mg/kg; Mg 23
2807 mg/kg). 22
The rabbit manure had the highest nitrogen 21
t °C
20
content (3.92%) compared to other manure, and 19
the other chemical elements in it were also high 18
17
in content. 16 17,45
Horse manure was low in nitrogen (1%) and 15
respectively low C/N ratio (44.50%), probably
due to the rough feed fed to the horses.
Table 2. Available animal waste and their chemical

characteristics Figure 2. Average daily temperature at ten days periods
Chemical elements
Type of рН
Season C N Р mg/ K
material (H2O)
% % kg mg/kg
Weather conditions affected the temperature of
Horse All year
7.55 44.50 1.00 0.68 1.45
the compost piles. The temperature graph of the
manure round
Cattle All year
spring compost piles shows several peaks,
manure round
7.45 35.56 2.08 0.88 0.65
which outlined the turning of the compost from
Pig
manure
All year
round
8.02 37.24 3.38 2.31 1.17 an initial mesophilic to a thermophilic and then
Mixed All year again into the mesophilic phase (Figure 3).
manure round
7.30 37.78 2.70 2.77 4.26 Composte pile V1 continuously passes from
(sheep
and goat) thermophilic to mesophilic phase after about 25
Rabbit All year
8.05 40.46 3.92 2.56 3.24 days, and after 27 days the temperature steadily
manure round
dropped below 30°C.
The thermophilic phase in compost V2 passes
Meteorological conditions during composting
after about 21 days, and after 24 days it
period were characterized by precipitation
dropped permanently below 30°C.
during the first and second ten days of June
(06-08 and 13-15.06.2016), with the sum of
precipitation being 49.4 and 23.1 l/m2
639
70 70
V1 V2
60 60 61
56
50 50
40 40
t °С
t °C
30 32
30
20 20
17
10 10 11
0 0
0 10 20 30 40 50 60 70 0 5 10 15 20 25
Days after starting Days after starting
Figure 3. Dynamics of temperature Figure 5. Changing the temperature

in compost piles in compost pile V3, bild in autumn
The fall compost was built in the second half of The last ten days of December, along with the
November. The weather conditions were fall of the snow, the temperature of the compost
appropriate - sunny weather without precipi- has dropped permanently and the temperature
tation and positive temperatures. During the measuring has been stopped (Figure 5).
first ten days of December, the air temperature
dropped several times below zero, creating a Table 3. Chemical composition of mature compost
risk of cooling the compost (Figure 4) Chemical elements
рН
Variants C N Р K Mg
(H2O)
8 (%) (%) mg/kg mg/kg mg/kg
V1 7.6 16.33 1,028 2311 4970 2451
6
7.5 12.65 1,005 2636 4434 2518
V2
4 V3 8.3 4.86 1,070 2029 3592 2301
2
At the end of May 2017, were taken samples
t °C
0
from the three compost piles for chemical
16.11.2017
21.11.2017
26.11.2017
01.12.2017
06.12.2017
11.12.2017
16.12.2017
21.12.2017
-2
analysis and tracking of composting processes
-4 (Table 3).
-6 The highest level of pH is compost V3. The
-8
content of phosphorus, potassium and
magnesium had lower values than the other two
Figure 4. Average daily air temperature composts. Compost V1 had the highest C and
for the period 20.11.-15.12.2017 K content, while compost V2 had the highest P
content, which may be due to the presence of
The temperature graph of the autumn compost rabbit fertilizer in the compost. In the calcu-
pile showed that the compost was self-heating lations of the ratio C/N at the beginning and
well and reached a high temperature of 61°C. end of the composting were found differences
The temperature curve clearly indicates that the in the ratio C/Nin the three compost piles.
compost passed through the mesophilic and There is a decrease in the C/N ratio, and for
thermophilic phases (Figure 5). spring compost pilesV1 and V2, this ratio has
With the fall of the compost temperature near changed in the range of 11-13:1, while in
10°C, which coincided with the negative autumn compost (V3) which at the beginning
temperatures during the first ten days of of the composting process is low , at the end of
December, there was a danger of a permanent composting is reduced to about 4.5:1.
cooling of the compost bundle. But the At this compost, all the basic materials showed
followed favorable daytime temperatures, along low C/N ratios, where brown materials were
with the large mass of plant mass, helped to virtually absent (Figure 6)
pass through (Figure 5).
640
ACKNOWLEDGEMENTS
30 25,53 26,28 V1 V2 V3
25 This research work was funded by Project №

14/19.01.2016: „Comparative study of different
20 types of compost for composting phases,
С/N
15 11,81 11,46 12,59 compost quality and application methods”, to

10 the Research Sector of the University of
4,54 Forestry. The financial support for participation
5
in the conference is under Project
0 BG05M2OP001-2.009-0034 „Support for the
starting end Development of Scientific Capacity at the
Figure 6. The C/N ratio of the compost piles University of Forestry”, funded by the
at the beginning of the composting process and Operational Program „Science and Education
in the mature compost for Smart Growth” (2014-2020).
A ratio of less than 20:1 at the end of REFERENCES
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The obtained amounts of the three compost Alami I., Soudi B., 2014. Dynamic composting
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sieved, from V2 (compost 2) 19.5 kg of Organic World Congress 2014, 13-15 Oct., Istanbul,
compost was sieved and from V3 (compost 3) Turkey.
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Visually, at the first two composts, there was Delgado A., López-Mosquera M E., 2012. Production
more waste from vine branches and fruit rods, and Characterization of Compost Made from Garden
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posting of vegetable waste for only 6 months. biological changes during composting of different
organic wastes and assessment of compost maturity.
CONCLUSIONS Bioresource technology, 96 (14), 1584-1591.
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of Composted Green Wastefor Horticultural Uses.
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M.C., Suárez-Estrella F., Jurado M., Moreno J.,
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Cultivation Journal of environmental science and Authors Food and Agriculture Organization of the
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Román P., Martínez M.M., Pantoja A., 2015. Farmer´s
Compost Handbook Experiences in Latin America
642
MORPHOLOGICAL AND ANATOMICAL INVESTIGATION

OF ALOYSIA CITRODORA PALAU - NEW MEDICINAL PLANT
INTRODUCED IN ROMANIA
Vasilica LUCHIAN1, Elena SĂVULESCU1, Jorj PĂDURARU1,

Vlad POPA2, Viorica LAGUNOVSCHI-LUCHIAN1
1
2
Research Center for Studies of Food Quality and Agricultural Products, USAMV Bucharest,
Corresponding author email: elena_să[email protected]
Abstract
Lemon verbena - Aloysia citrodora Palau (Verbenaceae) is an aromati plant used for the medicinal properties of
itsleaves and essential oils. The species is native to Argentina, Paraguay, Brazil, Uruguay, Chile, Bolivia and Peru. It is
cultivated and commercialised as an aromatic plant for its lemon-like scent of its leaves and flowers. The dried plant
and its extracts are valuable for medicinal preparations, in the perfumery industry and as an ingredient for the gourmet
cuisine.It is often used in phytotherapiessuch as a digestive and diuretic, antispasmodic, carminative and sedative,
antimicrobial and local analgesic. In 2015 Aloysia citrodora Palau was studied at theUniversity of Agronomic Sciences
and Veterinary Medicine of Bucharest. The study targeted the stems and leaves and it shows the presence of glandular
and non-glandular trichomeson all of its aerial parts. Morphological and anatomical analyses of the leaves and stems
were made with the plant material cultivated in our University at the Research Center for Studies of Food Quality and
Agricultural Products Greenhouse.
Key words: Aloysia citrodora, morphological, leaf structure.
INTRODUCTION indoors, a period in which the plant is leafless

and requires low amounts of water. In spring,
Aloysia citrodora, commonly called lemon the plant recovers with leaves. Aloysia
verbena is a native plant - to Argentina and citrodora has been a popular garden plant in
Chile. Aloysia citrodora Palau is synonymous warm southern and western parts of the U.S.
with Verbena triphylla L'Hér., Aloysia triphylla The plant has a gentle sedative action and
L’Hér., Lippia citriodora Kunth, and Lippia areputation for soothing abdominal discomfort,
triphylla (L’Hér.) Kuntze (Erikson, 2006). It is has amild tonic effect upon the nervous system
commonly known as “lemon verbena”, and helps to depression
Gattuso et al., 2008).
“cedrón”, “cidron”, “hierba Luisa” or The leaves of lemon verbena confer a
“verveineodorante”, according to the country. refreshing lemon flavor to the teas and
The genus Aloysia Ortega ex Jussieu belongs to infusions isused for digestive problems (as
the Verbenaceae. This genus comprises about flatulence, indigestion and acidity). The leaves
30 species, distributed on the American and flowers are used for culinary purposes
continent from the south of the United States (fruit salads and jams, teas, desserts),
down to Chile and Argentina. Spanish cosmetics, for potpourris and as herbal
explorers brought this plant to Spain in the 17th medicines (colds, fevers, dyspepsia and
century at which point it was named after diarrhea). In addition, the essential oil of lemon
Princess Louisa of Parma, genus name honors verbena has anti-bacterial and anti-fungal
Maria Louisa, princess of Parma and wife of properties (Hanna et al., 2011), while the
King Carlos IV of Spain. Lemon verbena isan essential oils, tea and tinctures proved an
evergreen plant in tropical or warm winter antibacterial and antioxidant activity (Cowan
locations, but deciduous in areas where 1999; Valentão et al., 2002; Sartoratto et al.,
freezing temperatures occur. In colder climates 2004; Pereira and Meireles, 2007).
it is grown in containers and overwintered Traditionally, lemon verbena is used as folk
643
remedy in treatments of spasms, cold and fever The fresh material, leaves and stems, whichwas
(Carnat et al., 1999), insomnia and anxiety used for the macroscopic and micro-
(Van Hellemon, 1986; Newal et al., 1996; morphologic studies, was collected in 2017, in
Wanmacher et al., 1990 a, 1990 b) and as the summer period, from the greenhouse of the
source of analgesic, antiinflammatory and/or Research Center for Studies of Food Quality
antipyretic remedies (Pascual et al., 2001). It is and Agricultural Products, located in
also often included in phytomedicines such as Bucharest, Romania.
digestive and diuretic (Duke, 1985; Torrent The cross sections intended to illustrate the
Martí, 1985), antispasmodic (Torrent Martí, morphological and anatomical aspect were
1985), carminative and sedative (Alonso Paz et made on the same material, in Botanics
al., 1992; Mors et al., 2000), antimicrobial and laboratory. The sections were clarified with
local analgesic (Hieronymus, 1882; Dellacasa chloral-hydrate for 24 hours, and stained with
and Bandoni, 2003). Used alone or in Alaun-Carmine and Iodine Green suitable to
combination with other herbs, the leaves make optical microscopy techniques. Observations
an excellent herbaltea. The essential oil is used and images of the anatomical structures in stem
in aromatherapy, extensively used in perfu- and leaves were made with the optical
mery, especially in making eau de toilette and microscope Leica DM1000 LED, Camera
eau de cologne. The plants repels midges, flies video Leica DFC295 and the Stereomicroscop
and other insects (Benzi et al., 2009). The Leica S8 APO, belonging to the Laboratory of
essential oil is an effective insecticide in 1-2% microscopy and plant anatomy of the same
concentration. Aloysia citrodora is codified by research center.
the Argentine Pharmacopeia (1978), Francaise
Pharmacopeia (1996), Argentine Alimentary RESULTS AND DISCUSSIONS
Codex (1969-2007) and European
Pharmacopoeia (2007). It is included in the Macroscopical characteristics. The lemon
FDA's GRAS list, i.e. the list of food additives verbena is a woody shrub, with lanceolate
which are Generally Regarded As Safe (Newall green leaves, with strong aroma and lemon-
et al., 1996). taste. The analysis of the foliar architecture
shows simple pinnate, leaves, 5-10 cm long,
MATERIALS AND METHODS lanceolate, with short petioles, in whorls of
three. The color of lamina is pale green, white
Plant material to pale lilac flowers, usually assembled in
The plant material is a species and not a terminal racemose inflorescences.
cultivar. The plant material to start the study Microscopical characteristics. The blades of
was bought from plant-shop.ro as a 15 cm plant the leaves exhibit anomocytic stomata on the
(initially in a jiffy peat pellet) in a 9 cm pot. abaxial side (Figure 1) and: two types of
In the spring of 2015 Paduraru Jorj began the trichomes present on both sides of the leaf
study of multiplying Aloysia citrodora. The surfaces, non-glandular and glandular
activity took place in the Hortinvest Green- trichomes (Figure 2, Figure 4). The dorsiventral
houses and in Domnesti village (44° 24′ 1.5′′ N, mesophyll, epidermic abaxial cells present
25° 54′ 55.1′′ E) Ilfov, 14 km of Bucharest. The striate cuticule around of the stomata. Metcalfe
crop was fertilised with BioHumusSol. and Chalk (1972) mentioned for the genus
Being the first experimental study of having Aloysia the occurrence of anomocyticstomata.
Aloysia citrodora in Romania's climate, the Mesophyll is dorsiventral (Figure 3), with a 2-3
study began with 200 plants in the field. layer of palisade parenchyma and spongy
Considering the results of testing the planting parenchyma cells, this tissue is located next to
distances of Aloysia citrodora in other tempe- the abaxial epidermis, the midrib consists of a
rate climates, such as in the experimental fields collateral vascular bundle and the lower
in Zalec, Slovenia, 1997, it was chosen the epidermis is unstratified and presents stomata.
40/40 cm as planting distance. At planting Transverse sections of the leaf measuring
distances in the field of 40 x 40 cm, the average 10.297 to 12.318 μm and palisade parenchyma
yield was 1050-1150 kg of dry leaves per measuring 3.610 to 4.404 μm (Table 1).
hectare.
644
Table 1. Leaf and palisade parenchyma thicknesses
Thick of leaf (µm) Palisade parenchyma (µm)
12.065 3.770
12.206 4.304
11.306 4.292
10.638 3.859
11.306 3,398
12.318 4.404
11.430 3.705
10,297 3.610
10.915 4.224
10.407 3.871
Figure 3. Leaf (cross section): e.ad. - adaxial

epidermis; p.p. - palisade parenchyma; b. - colateral
vascular bundle; p.s. - spongy parenchima;
e.ab - abaxial epidermis
Figure 1. Abaxial epidermis with anomocytic stomata
and glandular capitata trichomes
Figure 2. Adaxial epidermis

with non-glandular trichomes
Figure 4. Detail of abaxial epidermis

with non-glandular trichomes
645
Stem anatomy: the stem, in incipient second- reported the presence of three types of
dary growth, presents epidermis, angular glandular and one typeof non-glandular
collenchyma and the vascular bundles which trichomes.
are open collaterally; transverse sections show
a circular margin with 6 ribs, the epidermis is
unstratified, with stomata.
At the level of the ribs and in a subepidermal
position, 3-4 layers of angular collenchyma can
be observed and cork usually appears near the
phloem. Vascular bundles are open collaterally
(Figures 5 and 6).
Nonglandular and glandular trichomes were
observed in the unistratified epidermis as
described for the leaf.
For the Verbenaceae family several descrip-
tions of trichomes exist (Cantino, 1990;
Yashodhara et al., 2001). Specifically for A. Figure 5. Stem (cross section):
citrodora, Casadoro and Rascio (1982) in their x. - xylem; p. - parenchyma
ultrastructural study of its leaf trichomes,
Figure 6. Stem (cross section): e. – epidermis; ng.t. - non-glandular trichome; g.t. – glandular trichome;
co. - cork; c - colenchyma; s. – sclerenchyma; b. – bundle; x. - xylem
646
OBSERVATIONS IN THE FIELD. REVIEW From the morphological point of view there
The species does well in Romania mainly as an were determined: simple, entire, lanceolate,
annual crop. It showed that it reaches it's petiolate, pinnate leaves. The stem presented
biological potential, producing leaves above the many ribs.
the parameters mentioned in the morphology of After this study it can be mentioned the
the species (20.0 cm2 ): 10/2.5cm, 21.75 cm2 following anatomical characteristics of the
leaf. leaves: adaxial epidermis with anticlinal thin
For mechanized harvesting, it can be planted in walls and strains, abaxial epidermis with
wide strips. The recommended plant material is anticlinal thin and sinuous walls, anomocytic
vigorous rooted cuttings in a perlite and peat stomata, non glandular and glandular tricho-
mix substrate, using rooting hormones from mes, hypostomatic dorsiventral mesophyll. The
parent plants grown in the greenhouse. above described elements should be useful for
The species can be cultivated successfully in corect botanical identification of A. citrodora
Romania as an annual crop, where it has species.
demonstrated that it reaches its biological
vegetative potential, producing leaves beyond REFERENCES
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648
DYNAMICS OF MICROBIOLOGICAL INDICATORS

FOR COMPARATIVE STUDY OF COMPОST VARIANTS
Boyka MALCHEVA1, Milena YORDANOVA2, Radoslav BORISOV2,

Teodora VICHEVA3, Maya NUSTOROVA1
1
University of Forestry, Faculty of Forestry, 10 Kliment Ohridski Blvd, 1797, Sofia, Bulgaria,
2
University of Forestry, Faculty of Agronomy, 10 Kliment Ohridski Blvd, 1797, Sofia, Bulgaria,
3
University of Forestry, Faculty of Ecology and Landscape Architecture, 10 Kliment Ohridski Blvd,
1797, Sofia, Bulgaria
Abstract
The microbiological dynamics of different starting vegetable and animal waste was investigated. There is a 5 variants
scheme of compost bunches (4 spring and 1 autumn) with different starters - two spring composts containing a
combination of grape vine canes, fruit twigs and grass swath, in addition to one rabbit manure; starter: last year`s
compost, two spring composts of mulberry twigs with leaves, and when the compost are turned, a different amount of
clean litter and twigs is introduced; starter: soil and one autumn compost containing pepper and tomato stems and
leaves, roots of tomatoes and pepper with soil, green leaves of leeks, corn and leek waste. A 6-fold microbiological
analysis was started on the 7th day of the compost materials and repeated in 7-10 days during the compost reversal
period. Main groups of heterotrophic microflora - ammonifying bacteria (non-spore and bacilli), actinomycetes and
micromycetes are defined. The study was carried out by the method of dilution and culture of solid nutrient media with
determination of cfy (colony forming units) in 1 g abs. dry substrate. The data from the microbiological analysis show
differences in the course of the individual stages of composting by microbiological indicators. These differences are
expressed in terms of both the total amount of microorganisms in the substrates and the dominant physiological and
systematic groups of microorganisms in the microbiocenosis. The different microbiological composition of the compost
materials determines a different rate of decay of the separate raw materials, which is reflected in the duration of the
composting process itself.
Key words: compost, grape vine canes, horticulture wastes, microbiological dynamics, rabbit manure.
INTRODUCTION bacterial groups at the start of the composting

process are mesophilic bacteria producing
Microorganisms play an important role in the organic acids such as Lactobacillus spp. and
composting process. They use organic matter as Acetobacter spp., and later, in the thermophilic
a source of nutrients (Rynk et al., 1992; Borken stage dominated Gram-positive bacteria such as
et al., 2002), as a result of the development and Bacillus spp. and Actinobacteria (De Bertolli et
the activity in the formation of compost al., 1980). According to Abu-Bakar (2015),
removed heat, CO2, water vapor and forming a exactly bacteria provide the fastest and most
humus ( Epstein, 1997). Composting involves efficient composting. In his study, Partanen et
different types of microorganisms - bacteria, al. (2010) found that bacteria from
fungi and actinomycetes. They have different Actinobacteria, Bacteroidetes, Firmicutes,
characteristics and functions that are vital to the Proteobacteria, Deinococcus-Thermus and
process of composting (Lee, 2016). According from over 2000 different philotypes
Tiquai (2005) in addition to studies of participated in the composting process. Fungi
microbial biomass, respiration rate and content develop both in mesophilic and thermophilic
of ATP, the enzyme activity is also one of the phase of the composting, in an amount between
most effective methods that can be used to 0.01 and 1 million per gram compost (Kowalik,
monitor the stability and maturity of compost. 2015), as are present on the surface of the
In a typical composting process both bacteria compost, when the temperature is higher. Some
and fungi are present (Gray et al., 1971). types of actinomycetes develop during the
Earlier studies have shown that the main thermophilic phase, while others develop in
649
subsequent stages of maturation and ripening, bedding for silkworm growing and twigs from
by degrading resistant compounds in the final the same polluted area.
stages of humification (Lee, 2016; Adani et al., Microbiological studies include determination
1997; Andrews and Hirano, 1991). of non-sprouting bacteria, bacilli and
Successful composting depends on a number of micromycetes by method of selective plating
factors that have a direct and indirect impact on and direct viable counts. They were used two
the activity of microorganisms. Tiquia et al. solid nutrient media (meat-peptone agar for
(2000); Fracchia et al. (2006) indicate as determination of non-sprouting bacteria and
important factors the type of composite raw bacilli, and medium of Chapek-Dox for
material, its nutrient composition and its determination of micromycetes) and counting
physical characteristics such as bulk density, of colony forming units, recalculated to 1 g of
pH, moisture content, etc. According to Garcia absolute dry substrate.
et al. (1993) and Serra-Wittling et al. (1996), The statistical processing of microbiological
composting results in homogeneity in the data includes calculation of an average of three
composition of the product, irrespective of the replicates and a coefficient of variation.
starting material and the composting process
itself. RESULTS AND DISCUSSIONS
Biddlestone and Gray (1985) report that the
complexity of the degraded plant materials and Dynamics of compost temperature and pH
the quality of the final product may depend on For the determination of the composting
the type of biomass. In composting, the C: N phases, the temperature measurement started
ratio is considered the most important parame- with the building of the compost piles and
ter as it reflects the degree of biotransfor- continued until their last turning (Figure 1).
mations that have occurred in compost from a
chemical point of view (Saber et al., 2011). A 70
V1 V2 V3 V4
ratio of C: N of less than 12 during the solid 60
phase is an indicator of maturity of the compost 50
(Bernal et al., 1998; Iglesias-Jimenez, 1993). 40
The purpose of the present study is to trace the
t °C
30
dynamics of different compost variants on the 20
quantity and quality composition of 10
microorganisms developing in composts. 0
0 10 20 30 40 50 60 70
MATERIALS AND METHODS Days after starting
Figure 1. Dynamics of temperature in compost piles

There has been developed a 4 variants scheme
of compost piles (V1, V2, V3, V4) with The thermophilic phase of compost pile V1
different row materials: lasts for about 25 days, and after 27 days the
V1-Brown materials: grape vine canes and fruit temperature steadily drops below 30°C. The
twigs; Green materials: grass swath; Starter: thermophilic phase of compost pile V2 lasts for
last year compost residues. about 21 days, and after 24 days it drops
V2-Brown materials: grape vine canes and fruit permanently below 30°C. For compost pile V4
twigs; Green materials: grass swath and rabbit the thermophilic lasts for about 13 days, and
manure; Starter: last year compost residues. after about 26 days, the temperature is
V3-Mulberry twigs with leaves; Starter - soil. continuously dropping below 30°C.
On the 7th and 13th day of building the compost The compost pile V3 did not switch to a ther-
pile, when it was turning, were added bedding mophilic phase. The highest measured
for silkworm growing and twigs. temperature in this pile is 34°C, reached around
V4-Mulberry twigs with leaves (from a the 13th day and, similar to V4 compost, after
contaminated area with heavy metals); Starter - about 26 days, the temperature steadily drops
soil. On the 7th and 13th day of building the below 30°C.
compost pile, when it was turning, were added
650
By making a comparison of the temperature at (V1), plant wastes have the highest microbial
different compost piles, with the amount of presence, whereas variants with a different
material used for their building, it can be seen amount of mulberry leaf and twigs (V3 and V4)
that from the four composting piles, with the have a lower microbial diversity and presence.
highest temperatures and longest lasting Reducing the amount of started plant waste
thermophilic phase is compost pile V1, which materials and adding the rabbit fertilizer to V2
is with the most greater mass of starting slows the growth of microorganisms at the start
material (59,500 kg). In descending order, for of the experiment (Day 7), but in the next days
this indicator are arranged compost pile V2 of reporting, their activation is determined,
(56,050 kg) and compost pile V4 (32,200 kg). most preferably on the 13th day.
Compost pile V3, which has the smaller mass
of the starting material (24,330 kg), did not 20000000
V1 V2 V3 V4
reach the thermophilic phase.
total number of microbes

Throughout the composting process, the pH of 15000000
the compost piles was monitored and it was.

10000000
between 7.8 and 8.0. During the thermophilic
phase and the maturation phase, normally the
5000000
medium is alkaline. Probably because of the
rapid passage from mesophilic to thermophilic 0
phases, no acidification of the medium from the 2 7 13 22 29 39 48
microbial extraction of organic acids in the Days after starting
mesophilic phase was established. According to
Figure 2. Dynamics of total microflora
some authors, low initial pH limits microbial
(number of microbes/g substrate)
activity and slows temperature rise (Sundberg
et al., 2004; Romanschuk et al., 2005).
In V2, the amount of the total microflora
Microbiological analyses decreases more gradually, whereas in the other
The microbiological analyses were started one variants, the multiplication of microorganisms
week after the building of compost piles V1, decreases sharply to the 13th day - 2 times at V1
V2 and V4, and on the 2nd day after the and V3, and 3 times at V4, after which the
building of compost pile V3. The analyzes breeding process gradually decreases to the end
were repeated at each compost turn (between 7- of the composting, with a slight increase in
10 days). microbial growth on the 29th day after the trial.
The results of the dynamics of the total In all four composting variants it was found
microflora (total number of microbes in 1 g of that on the 39th and 48th day the amount of
substrate) give an idea of the degree of microbes was the lowest. At the end of
development, resp. settlement of the compost composting, it decreased about 2 times for
with microbes. This indicator is important for variants V1, V2 and V3, and 3 times for V4
assessing the degree of destruction of organic compared to the amount at the beginning of the
wastes as far as the microorganisms carry out experiment. By the seventh day of the
the mineralization of the organic compounds in experiment, mesophils and thermophiles are
them. Data from the general microflora grown at V1, V2 and V4, whereas at V3 were
dynamics are presented in Figure 2. found only mesophils, as this compost does not
The data show a different dynamics of the total pass through the thermophilic phase. In the
microflora in the four composting variants. It is period from 22 to 29 days, temperatures rise
also possible to note the different start for the above 60°C at V1 and above 40°C at V2 and
presence of microorganisms in the individual V4, i.e. these variants pass through the
variants provided by the various formulations. thermophilic phase again. The most active is
On the 7th day of experimentation, the highest the development of microorganisms up to the
amount of total microflora was found at V1. 7th day of experiment. Activation of
This amount is about 1 times higher than the thermophiles and increase of total microflora
same in the other variants. For this variant from Day 22 to Day 29 of experimental was
set-up - 1 time at V1, 1, 2 times at V2, 1, 3
651
times at V4. The most active is the stress factor for the development of
development of mesophils in V3 during this microorganisms which limits their development
period - the total amount of microorganisms is and changes the role of the different groups of
increased by 1, 5 times. After 29 days to 48 microorganisms in compost 1 and 3. A major
days, compost temperatures are below 30°C, share in the composition of the total microflora
i.e., mesophilic microorganisms develop, the again occupies non-sprouting bacteria and
composts are aged and ripened. bacilli, with the exception of composts 1 and 3,
In the different phases of composting, different where the development of micromycetes is
quantitative development of the studied more active than that of the bacilli. Compost
microbial groups - non-sprouting bacteria, variants are in the mesophilic phase,
bacilli and micromycetes is also reported. The decomposition of organic matter occurs with
results for the composition of the microflora for the development of mesophils.
7 days (passage through the mesophilic and The composition of the microflora 22 days after
thermophilic phase) after the experimental the trial of the experiment is presented on Table
assay in all variants and on the 2nd day (meso- 3:
philic phase) at V3 are presented in Table 1.
Table 3. Microflora composition 22 days after the trial
Table 1. Composition of the microflora 7 days after the (CFU × 103/g compost) ± CV (%)
trial (CFU × 103/g compost) ± CV (%) Variants Non-sprouting Bacilli Micromycetes
Variants Non-sprouting Bacilli Micromycetes bacteria
bacteria 5600 ± 0.089 520 ± 0.385 960 ± 0.208
V1
(79.1) (7.3) (13.6)
V3 13760 ± 0.262 1720 ± 0.291 100 ± 0.400
4800 ± 0.208 500 ± 0.400 1560 ± 0.385
(2-ри ден) (88.3) (11.0) (0.6) V2
(70.0) (7.3) (22.7)
13120 ± 0.202 2940 ± 0.340 60 ± 0.333
V1 4000 ± 0.250 1900 ± 0.316 200 ± 0.250
(81.4) (18.2) (0.4) V3
(65.6) (31.1) (3.3)
11200 ± 0.179 2400 ± 0.300 80 ± 0.125
V2 3040 ± 0.164 1460 ± 0.342 500 ± 0.100
(81.9) (17.5) (0.6) V4
(60.8) (29.2) (10.0)
10880 ± 0.243 1720 ± 0.116 140 ± 0.357
V3
(85.4) (13.5) (1.1)
13600 ± 0.074 2060 ± 0.243 90 ± 0.222
The composition of the total microflora follows
V4
(86.3) (13.1) (0.6) the same trend on the 29th day of the
experiment - a higher amount of ammonifiable
Mesophils and thermophiles develop in these bacteria and lower micromucites (Table 4).
two phases - mesophilic and thermophilic, with
the highest contribution to the composition of Table 4. Microflora composition 29 days after the trial
the general microflora as non-sprouting (CFU × 103/g compost) ± CV (%)
bacteria, followed by the bacilli, which are Variants Non-sprouting Bacilli Micromycetes
bacteria
actively involved in the initial stages of 6000 ± 0.200 920 ± 0.217 260 ± 0.385
destruction of organic matter. V1
(83.6) (12.8) (3.6)
The microbiological analyses at the 13th day V2
5880 ± 0.307
(71.4)
1940 ± 0.258
(23.5)
420 ± 0.238
(5.1)
(mesophilic phase) after experimentation show 7920 ± 0.126 780 ± 0.256 560 ± 0.107
V3
a different quantitative development of the (85.5) (8.4) (6.1)
5200 ± 0.192 520 ± 0.288 600 ± 0.167
different groups of microbes (Table 2): V4
(82.3) (8.2) (9.5)
Table 2. Composition of the microflora 13 days The temperatures of the compost piles in the
after the experiment (CFU × 103/g compost) ± CV (%)
two last samples are close - ranging from 20°C
Variants Non-sprouting Bacilli Micromycetes to 26°C.
bacteria
5120 ± 0.195 820 ± 0.244 1020 ± 0.196
On the 39th day after experimentation, non-
V1
(73.6) (11.8) (14.7) sprouting bacteria and bacilli dominate, with
5200 ± 0.254 960 ± 0.417 4000 ± 0.150
V2
(51.2) (9.4) (39.4)
the exception of compost 2, where the amount
V3
4940 ± 0.202 880 ± 0.341 240 ± 0.417 of micromycetes is higher than that of bacilli
(81.5)
4040 ± 0.248
(14.5)
880 ± 0.455
(4.0)
480 ± 0.208
(Table 5).
V4
(74.8) (16.3) (8.9) As on the 39th day and 48th day of experi-
mentation, the temperature range suggests the
The lower temperature with 7°С (V1), 13°С
development of mesophilic microorganisms.
(V2), 5°С (V3) and 18°С (V4) for 1 week is a
652
For all compost materials, the amount of non- table materials. The lowest in the composition
sprouting bacteria and bacilli is higher than that of the total microflora is the micromycetes.
of micromycetes (Table 6).
ACKNOWLEDGEMENTS
Table 5. Composition of the microflora 39 days
after the trial (CFU × 103/g compost) ± CV (%) This research work was funded by Project №
Variants Non-sprouting Bacilli Micromycetes 14/19.01.2016: „Comparative study of different
bacteria types of compost for composting phases,
5160 ± 0.194 800 ± 0.250 20 ± 0.500
V1
(86.3) (13.4) (0.3) compost quality and application methods”, to
V2
4840 ± 0.179 500 ± 0.200 1160 ± 0.086 the Research Sector of the University of
(74.5) (7.7) (17.8)
3960 ± 0.253 460 ± 0.326 400 ± 0.125
Forestry. The financial support for participation
V3
(82.2) (9.5) (8.3) in the conference is under Project
3760 ± 0.266 760 ± 0.066 260 ± 0.154
V4
(78.7) (15.9) (5.4)
BG05M2OP001-2.009-0034 „Support for the
Development of Scientific Capacity at the
Table 6. Microflora composition 48 days University of Forestry”, funded by the
after the trial (CFU × 103/g compost) ± CV (%) Operational Program „Science and Education
Variants Non-sprouting Bacilli Micromycetes
for Smart Growth” (2014-2020).
bacteria
V1
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654
CONTRIBUTIONS TO HALOPHILIC FLORA

AND VEGETATION IN OLTENIA (ROMANIA)
Daniel RĂDUȚOIU1, Cătălin George SIMEANU1, Ion STAN2

1
University of Craiova, Faculty of Horticulture, Department of Biology and Environmental
Engineering, 13 A.I.Cuza Street, 200583-Craiova, Romania
2
University of Craiova, Faculty of Agriculture, 15 Libertăţii Street, 200583-Craiova, Romania
Abstract
The halophilic flora from Oltenia is known only from few localities: Gighera, Seaca de Câmp, Bratovoiești, Sadova,
Tâmburești, Piscu Sadovei, Murta, Dobrești, Afumați (Dolj County), Ocnele Mari - Ocnița (Vâlcea County), Osica de
Sus, Gura Padinii (Olt County).
By identification of new surfaces with halophilic plant species inside Oltenia region (Rastu Vechi - Dolj County) are
brought important contributions regarding the chorology of these plant species, associations and habitats. The
halophilic vegetation is fragmentary present near springs and small streams with salt water. From the habitats with
halophilic vegetation in Romania on the area investigated by us we find: R1511 West-Pontic communities with Crypsis
aculeata, R1514 West-Pontic communities with Trifolium fragiferum, Cynodon dactylon and Ranunculus sardous,
R1521 Pontic-Sarmatic communities with Puccinellia limosa and Plantago maritima, R1529 Pontic-Pannonian
meadows with Hordeum hystrix and R1507 Pontic-Pannonian meadows with Carex distans, Taraxacum bessarabicum
and Aster tripolium ssp. pannonicus. Some of these habitats have a high conservative value (ex. R1521) while others do
not know area in this country side (ex. R1507).
Key words: flora, Oltenia, Romania, salty area, vegetation.
INTRODUCTION Following the consultation of specialty

literature which has as study object the
The origin of the salty fields from Oltenia are halophilic flora and vegetation inside Oltenia
either from salt deposits (in case of those from area (Buia et al., 1959, 1960, 1961; Șerbănescu,
Ocnele Mari - Ocnița area - Vâlcea County) or 1963; Păun et al. 1971; Popescu et al., 2000;
climate consequences, in which the Răduțoiu, 2013, 2014) and from country (Doltu
efflorescence processes or slightly soluble salt et al., 1979; Mihai, 1969; Pop, 1968; Prodan,
exudation, predominates over leaching 1922; Sanda and Ciobanu, 1967; Șerbănescu,
processes (Țopa, 1954). 1963; Țopa, 1939, 1954, 1965; Dihoru, 1990)
The salty lands are easily distinguished from we found out that in this side of country are
distance by the lighter color compared to the few data (Popescu et al., 2000; Răduțoiu, 2013,
normal soil in that area. The most widespread 2014, 2017).
salty lands in our area belong to solonceac or Almost 40 years ago, Al. Buia et al. (1961),
soloneț types without sodium carbonate (Țopa, reported in this area, in the meadows, on low
1954; Popescu et al., 2000). salty soils, numerous species such as: Trifolium
The presence of salty lands lacking vegetation ornithopodioides, T. subterraneum, T.
is due to missing air from soil. echinatum, T. angulatum, Medicago arabica,
The classification of halophilic plant species is M. hispida and Scorzonera cana
different realized from one author to another. (Podospermum canum).
Some herbalists (Țopa, 1954; Popescu et al., However, in the consulted research papers,
2000) are grouping the plant species in 4 big were not found any data regarding the
categories: mandatory, preferably, tolerant and halophilic flora and vegetation from south part
accidental, others (Prodan, 1939) in first of country between Rastu Vechi and Rastu Nou
category, second and third. localities. In the IX volume from Romania
Flora is mentioned a point for Aster tripolium
655
L. ssp. pannonicus (Jacq.) Soó (between also by the aesthetics given by the color of
Negoiu and Catanele) (Morariu and Nyárády, these plant species flowers. In the lilac sea
1964 in Săvulescu et al.). The identification of offered by Aster tripolium ssp. pannonicus
appreciable surfaces with this type of (Figure 1) we find a yellow winding carpet
vegetation made us to take in study this for given by Lotus tenuis to which fixation
having a real situation and to propose the participates species like Trifolium fragiferum
protection of these places being known that the ssp. bonannii and T. repens, being speckled by
surfaces occupied by this kind of flora and their reddish and white flowers.
vegetation in Oltenia region are very few.
The territory with halophilic vegetation
between Rastu Vechi and Rastu Nou is near the
Danube (N43º54'690"; E23º17'161", Alt. 52
m.s.m.). We can say that is placed on the upper
terraces of the Danube. It is a flat field with a
length of almost 1875 m and a variable width,
between 3.75 m and 40 m, totaling over 4 ha.
For making this study were made numerous

field trips starting from 2006, when these salty
fields appeared for the first time, after the Figure 1. Aster tripolium ssp. pannonicus between Rastu
Vechi and Rastu Nou localities
catastrophic floods from the spring of same
year to the present. The collected data were On the background of these colors we meet
analyzed and compared with those already clusters of grasses which diversifies the
present from other stations with halophilic chromaticity of the place (Puccinellia limosa,
vegetation from Oltenia. Crypsis aculeata, C. schoenoides etc.).
The samples that cannot be identified in the The floral list of salty lands from Rastu Vechi-
field were collected using special material, Rastu Nou area totals a number of 71 taxa:
specific to this study and subsequent identified Phylum Spermatophyta, Magnoliopsida Class,
using the specialty work papers from our Ranunculaceae Family: Ranunculus sardous
country and abroad (Beldie, 1977, 1979; Cr., R. sceleratus L., Consolida regalis S.F.
Ciocârlan, 2009; Sârbu et al., 2013; Săvulescu Gray, Caryophyllaceae Family: Cerastium
et al., 1952-1976; Tutin et al., 1964-1980). dubium (Bast.) Guépin, Gypsophila muralis L.,
After identification, the material was preserved Spergularia rubra (L.) J&C Presl.,
in the botanical lab to be included in the Chenopodiaceae Family: Atriplex patula (L.),
herbarium from University of Craiova (CRA). A. prostrata Boucher ex DC., A. tatarica L.,
The authors’ abbreviations were done Chenopodium glaucum L. Polygonaceae
according to Brummitt and Powell (1992). Family: Polygonum aviculare L., P.
After establishing the existing vegetation type arenastrum Boreau, Rumex conglomeratus
in the research area was made also a framing of Murray, R. crispus L. Rosaceae Family:
it to the Romania habitats (Doniță et al., 2005; Potentilla reptans L., Fabaceae Family: Lotus
Gafta and Mountford, 2008). tenuis Waldst. & Kit., Medicago lupulina L.,
The coordinates were noticed using a Garmin Melilotus albus Medik., M. dentatus (Waldst.
etrex 30 GPS. & Kit.) Pers., Trifolium fragiferum L. subsp.
bonannii (C. Presl) Soják, T. repens L.
RESULTS AND DISCUSSIONS Lythraceae Family: Lythrum virgatum L.,
Apiaceae Family: Bupleurum tenuissimum L.,
Following analysis of the surfaces occupied by Peucedanum latifolium (Bieb.) DC., Malvaceae
this type of vegetation in the research territory, Family: Althaea officinalis L., Brassicaceae
we can say that the salty lands from here are Family: Cardaria draba (L.) Desv., Diplotaxis
interesting not only because of plant species but muralis (L.) DC., Lepidium perfoliatum L., L.
656
ruderale L., Rorippa austriaca (Cr.) Bess., R. xerophilic and mesophilic species can be
sylvestris (L.) Bess. ssp. kerneri (Menyh.) Soó, explained by the periods of drought present in
Sisymbrium polymorphum (Murray) Roth, this territory during the summer.
Gentianaceae Family: Centaurium pulchellum Table 1. The analysis of moisture index
(Sw.) Druce, Boraginaceae Family:
Nr. crt. Scale for moisture Nr. taxa
Heliotropium supinum L., Scrophulariaceae
Mezohigr. 27
Family: Gratiola officinalis L., Verbascum
1. Mez. 8
blattaria L., Lamiaceae Family: Mentha 2. Xeromez. 7
pulegium L., Plantaginaceae Family: Plantago 3. Mezohigr.-higr. 6
cornuti Gouan, P. uliginosa F.W. Schmidt, 4. Higr. 6
Asteraceae Family: Achillea collina Becker ex 5. Mez.-mezohigr. 3
Rchb., Aster tripolium L. ssp. pannonicus 6. Xeromez.-mez. 3
(Jacq.) Soó, Bidens cernua L., Inula britannica 7. Xeromez.-mezohigr. 2
8. Mezoxer. 2
L., Lactuca saligna L., Matricaria recutita L.,
9. Mez.-higr. 2
Pulicaria vulgaris Gaertn, Scorzonera cana 10. Eurif. 2
(C.A. Mey.) Griseb., Sonchus arvensis L. ssp. 11. Higr.-hidr.. 1
uliginosus (M. Bieb.) Nyman, Taraxacum 12. Xer.-xeromez. 1
bessarabicum (Hornem.) Hand.-Mazz., 13. Xer. 1
Liliopsida Class, Butomaceae Family: Butomus
umbellatus L., Juncaceae Family: Juncus
bufonius L., J. compressus Jacq., Cyperaceae
Family: Bolboschoenus maritimus (L.) Palla,
Carex distans L., C. divisa Huds., Poaceae
Family: Agrostis stolonifera L., Alopecurus
pratensis L., Bromus commutatus Schrader,
Crypsis aculeata (L.) Aiton, C. alopecuroides
(Pill. & Mitterp.) Schrad., C. schoenoides (L.)
Lam., Cynodon dactylon (L.) Pers., Elymus
repens (L.) Gould, Festuca arundinacea
Figure 2. The ecological spectrum of salty lands plant
Schreb., F. pulchra Schur, Hordeum species from the research area
geniculatum All., Lolium perenne L.
Phragmites australis (Cav.) Steudel ssp. As a general observation on these salty lands, is
humilis (De Not.) Asch. et Graebn., Poa the lack of the typical species for salty lands:
bulbosa L., Puccinellia limosa (Schur) Holmb., Camphorosma annua Pall. Salicornia europaea
P. convoluta (Horrnem.) Hayek ssp. L. and Suaeda maritima (L.) Dumort., aspect
pseudobulbosa (E.I. Nyárády) Borza. mentioned also on the salty lands from Seaca
The highlighting of ecological particularities of de Câmp (Popescu et al., 2000).
all plant species from the research area allows From the habitats with halophilic vegetation in
us to determine the ecological specificity of Romania on the territory researched by us we
vegetation in accordance with the complex of find: R1507 Pontic-sarmatic meadows of Carex
local pedoclimatic factors. distans, Taraxacum bessarabicum and Aster
The analysis was based on the moisture index tripolium ssp. pannonicus, R1511 West-Pontic
because it is the only climatic factor that could communities with Crypsis aculeata, R1514
be observed and appreciated in the field. West-Pontic communities with Trifolium
From its analysis it can be observed that first fragiferum, Cynodon dactylon and Ranunculus
place is occupied by the mesohigrophilic plant sardous, R1521 Pontic-sarmatic communities
species (Table 1, Figure 2) with almost 40%. If with Puccinellia limosa and Plantago
we add also the plant species that have an index maritima, R1529 Pontic-Pannonian meadows
of moisture close to that one specific to of Hordeum hystrix.
mesohigrophilic species, we can realize that The vegetation characteristic of these habitats
these salty lands belong to the category of those is framed to the next associations: Taraxaco
with high humidity. The presence of some bessarabici, Caricetum distantis Wendelberger
1943; Crypsidetum aculeatae (Bojko 1932)
657
Ţopa, 1939; Trifolio fragifero - Cynodontetum
Br.-Bl. et Balas 1958; Puccinellietum limosae
Rapaics ex Soó 1933, 1936 and Hordeetum
hystricis (Soó 1933) Wendelberger 1943.
Taraxaco bessarabici, Caricetum distantis
Wendelberger 1943
This is the association to which the
phytocoenosis are enlightened by the caespitose
plant species called Carex distans.
Next to it, a good representation has Aster
tripolium ssp. pannonicus. The cortege of
species is completed by Taraxacum
bessarabicum, Festuca pulchra, Atriplex Figure 4. Puccinellietum limosae - autumnal aspect
hastata, Trifolium repens, Trifolium fragiferum This is the vegetation characteristic to R1521
ssp. bonannii, Medicago lupulina and Cynodon habitat, that at national level has a high
dactylon. conservative value (Doniță et al., 2005).
The vegetation framed to this association is Trifolio fragifero - Cynodontetum Br.-Bl. et
characteristic to the R1507 habitat and has a Balas 1958
moderate conservative value. Is characteristic to R1514 habitat. Is recognized
Crypsidetum aculeatae (Bojko 1932) Ţopa through the nucleus of subhalophilic species:
1939 Cynodon dactylon, Lolium perenne,
The phytocoenosis of these association have Gypsophila muralis, Ranunculus sardous,
small plant species in composition. Next to the Consolida regalis and Atriplex prostrata.
dominant one (Crypsis aculeata - Figure 3) we Has a low conservative value.
meet Aster tripolium ssp. pannonicus, Hordeetum hystricis (Soó 1933) Wendelberger
Taraxacum bessarabicum, Crypsis 1943
schoenoides, Trifolium fragiferum ssp. The surfaces occupied by the phytocoenosis of
bonannii, Cynodon dactylon etc. this association are small (few m2) and placed
in area that become dry during the summer.
Next to the dominant species we find:
Cerastium dubium, Gypsophila muralis,
Plantago uliginosa, Matricaria recutita,
Lepidium ruderale, Achillea collina and rare
specimens of Puccinellia limosa.
It is characteristic to the R1529 habitat and has
a moderate conservative value.
In the territory researched by us the salty
vegetation in near the one framed to
Phragmitetea australis because is placed close
to a water channel flowing from Seaca de
Câmp to Negoi, settlements neighboring to
Figure 3. Crypsidetum aculeatae
from area investigated those in between are placed the researched salty
lands.
These characterize the vegetation of R1511 To the west part of the land are present
habitat, that at national level has a moderate numerous specimens of Phragmites australis
conservative value. ssp. humilis what makes a cover of 60-70%
Puccinellietum limosae Rapaics ex Soó 1933, (Figure 5). This thing let us to think that in the
1936 (Figure 4) - is found in those places with next future will be formed phytocoenosis of the
higher moisture. The most frequent halophilic association called Astero tripolii -
plant species in these phytocoenosis are: Lotus Phragmitetum humilis Krisch (1972) 1974
tenuis, Hordeum geniculatum, Trifolium association that characterizes the R5311
fragiferum ssp. bonannii and Carex distans. habitat. West-Pontic communities with
658
Phragmites australis ssp. humilis and Aster Buia Al., Păun M., Safta I., Pop M., 1959. Contribuții
tripolium. geobotanice asupra pășunilor și fânețelor din Oltenia.
Lucr. Șt. Inst. Agron. Tudor Vladimirescu. Craiova.
90 pag.
Buia Al., Păun M., Safta I., Pop M., 1960. Contribuţii
geobotanice asupra păşunilor şi fâneţelor din Oltenia.
Lucr. Şt. Inst. Agron. “T. Vladimirescu”: 93-180.
Craiova.
Buia A., Păun M., Maloş C., 1961. Pajiştile naturale din
Regiunea Craiova şi îmbunătăţirea lor (II). Probleme
Agricole. Anul XIII: 31-40.
Ciocârlan V., 2009. Flora ilustrată a României.
Pteridophyta et Spermatophyta. 1041 pag. Edit.
Ceres, Bucureşti.
Dihoru Gh., 1990. Über einige Limonium-arten der Flora
Rumäniens. Analele Univ. Bucureşti. Biol., XXXIX:
46-50.
Doltu M.I., Sanda V., Popescu A., 1979. Vegetaţia
Figure 5. Areas where Phragmites australis ssp. humilis solurilor saline şi alcaline din România. Muz.
has a good representation Brukenthal. Stud. Comun. Şt. Nat. 23: 197-219.
Doniţă N., Popescu A., Paucă-Comănescu M.,
Mihăilescu S., Biriş I.-A., 2005. Habitatele din
CONCLUSIONS România. 496 pp. Edit. Tehnică Silvică, Bucureşti.
Gafta D., Mountford O. (coord.), 2008. Manual de
interpretare a habitatelor Natura 2000 din România,
Comparing the flora from the territory Edit. Risoprint, Cluj-Napoca.
researched by us with the one of salty lands Mihai Gh., 1969. Cercetări asupra vegetaţiei halofile din
from other areas from Oltenia region, we can bazinul Başeului (jud. Botoşani). Stud. Comun. Muz.
say that the one from other stations is richer in Şti. Nat. Bacău: 129-140.
species of Trifolium (ex. T. angulatum, T. Morariu I., Nyárády E.I., 1964. Genul Aster L. In:
Săvulescu et al. Flora R.P.R. Edit. Acad. Române,
campestre, T. dubium, T. echinatum, T. București. Vol. IX: 187-212.
hybridum, T. ornithopodioides, T. pratense, T. Păun M., Georgescu L., Fulga G., 1971. Importante
retusum, T. resupinatum and T. striatum). puncte floristice şi de vegetaţie în cuprinsul Olteniei.
The influence of zoo-anthropogenic factor is Stud. Cercet. C.C.E.S. Dolj: 67-84. Craiova.
visible on the salty lands vegetation from Rastu Pop I., 1968. Flora şi vegetaţia Câmpiei Crişurilor.
Interfluviul Crişul Negru-Crişul Repede. 280 pp.
Vechi-Rastu Nou area by: their location in the Edit. Acad. Române.
vicinity of agricultural fields, collecting of Popescu G., Simeanu C., Costache I., Răduţoiu D., 2000.
some plant species by the locals (ex. Preliminary Data Regarding the Flora of Some Salty
Taraxacum bessarabicum) or by irrational Fields in Oltenia. Acta Horti Bot. Bucurest. 28: 89-
grazing with sheep or goats (rarely horses), that 96.
Prodan I., 1922. Ecologia plantelor halofile din România,
affects the appreciated feed plant species: în comparative cu cele din Ungaria şi şesul Tisei.
Trifolium fragiferum, Lotus tenuis, Puccinellia Bul. Grăd. Bot. şi al Muz. Bot. Univ. Cluj, 2 (1): 1-
limosa, Crypsis aculeata, C. schoenoides, Aster 36; 2: 38-68; 3: 69-84; 4: 101-112.
tripolium ssp. pannonicus. Prodan I., 1939. Flora pentru determinarea și descrierea
The need for protection of these surfaces with plantelor ce cresc în România. Tipografia Cartea
Românească, Cluj. 713 pag.
halophilic vegetation in Oltenia is justified not Răduțoiu D., 2013. West coast communities with
only by the reduced area in Oltenia, but also by Pholiurus pannonicus and Plantago tenuiflora from
the presence of some new habitats for this part Oltenia. Annals of the University of Craiova. Seria
of country (R1507) or with high conservative Biologie, Horticultură, Tehnologia prelucrării
value (R1521). produselor agricole, Ingineria Mediului. XVIII (LIV):
639-644.
Răduțoiu D., 2014. New data about Limonium
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660
PLANT GROWTH PROMOTING BACTERIA

WITH ANTIFUNGAL ACTIVITY
Oana-Alina BOIU-SICUIA1,2, Sorina DINU2, Cristina PETRIȘOR2,

Florica CONSTANTINESCU2
1
Faculty of Biotechnologies, 59 Mărăști Blvd, District 1, Bucharest, Romania
2
Research-Development Institute for Plant Protection, 8 Ion Ionescu de la Brad Blvd,
Abstract
The aim of the present study was to analyse the potential of two plant beneficial bacteria, Cp.b4 and 75.1s, to prevent
fungal pathogens proliferation and improve tomato seedlings growth. The bacterial strains were selected from the
RDIPP microbial collection, due to their antifungal activity towards solanaceous plant pathogens. Both strains were
previously identified as Bacillus cereus/thuringiensis based on their biochemical profile with the Biolog GEN III
system. Growth chamber tests on tomato seedlings have shown that biological treatments, with the above-mentioned
bacterial strains, induced a better vegetative growth to the seedlings, and increased the photosynthetic capacity of the
plants. To prevent early blight caused by Alternaria sp. in solanaceous plants, the ability to inhibit conidia germination
was analysed. Biological treatments with bacterial suspension reduced the number of germinated conidia by 4.98 fold
(when using 75.1s strain) and by 2.57 fold (when using Cp.b4 strain). The study also revealed that mixed treatments of
bacteria suspension along with a low dose of fungicide inhibit conidia germination, being comparable with the
chemical control, Mycoguard 500 SC, applied at the recommended dose of 0.2%.
Key words: PGPB, tomatoes, early blight prevention.
INTRODUCTION blossom end rot (calcium deficiency) (Chaerani

and Voorrips, 2006; Kennelly et al., 2012). It
Tomato culture is an important vegetable is, therefore, important to maintain a good
grown in greenhouses and open fields. The physiological status during plant growth in
culture has numerous pests and diseases in all order to prevent infections. Moreover,
stages of vegetation and post-harvesting. microbial inoculation with plant growth
Moreover, new races of pathogens and promoting bacteria having biocontrol activity
emerging pests make plant protection more could improve plants health and vigour in order
difficult. Therefore, there is a continuous to prevent pathogenic infections (Wang et al.,
interest in preventing pathogenic infections and 2008; Jagadeesh and Jagadeesh, 2009; Zahoor
increasing the productive potential of the plant. et al., 2017).
Among plant pathogens, Alternaria spp. (like Considering these, we studied two plant
A. solani, A. alternata or A. tomatophila) are beneficial bacteria, Cp.b4 and 75.1s, regarding
ubiquitous fungal phytopathogens, which can their biocontrol potential in preventing
induce early blight in various plant species, Alternaria sp. and other fungal pathogens
including tomatoes and other edible plants proliferation. Microbial ability to improve
(Thomma, 2003; Woudenberg et al., 2015). tomato seedlings growth was also evaluated.
These pathogens infect plants in various growth
stages, including vegetable fruits during MATERIALS AND METHODS
ripening, causing fruit rot. It survives and
overwinters as saprophyte on plant debris. The Bacterial strains
leaf infections occur on cool, wet weather, and Two bacterial strains were used in this study,
fruit rot symptoms are usually associated with a 75.1s and Cp.b4, both from the RDIPP
predisposing injury due to sunscald, frost, or microbial collection. These two strains were
661
isolated from soil of Bărăgan area (75.1s) and assay was performed against five plant patho-
onion rhizosphere from Dolj county (Cp.b4). gens, Alternaria sp. (new isolate from tomato
Bacteria were stored at -80°C, in Luria Bertani plant), Botrytis cinerea (new isolate from
(LB) medium supplemented with 30% glycerol. tomato fruit), Fusarium oxysporum f. sp.
Routinely, they were grown on LB agar, at radicis lycopersici ZUM 2407, Pythium
28°C. debaryanum DSM 62946 and Rhizoctonia
solani DSM 63002.
Bacterial identification procedure Bacterial ability to inhibit conidia germination
Bacteria were identified with the Biolog was also studied against Alternaria sp. Tests
GENIII system, using protocol B for Gram- were performed in vitro, in 2 ml tubes, on
positive, spore forming, fast growing bacilli, Potato-Dextrose-Broth. Fungal spore suspen-
according to the manufacturer’s guidelines (US sion was prepared in 105 conidia/ml, and
Patent 5,627,045). Therefore, each bacterial bacteria in 108 cfu/ml. A chemical control was
strain was grown on Biolog Universal Growth also included within the study. Mycoguard®
medium, at 33°C, for not more than 18 hours. 500SC commercial product, based on chlorot-
Isolated colonies were then homogenised in B halonil 500 g/l, was used at the recommended
type inoculation fluid, at 97% turbidity, concentration (0.2%) for Alternaria spp.
detected in 590 nm wavelength light. Biolog control (according to the pesticide factsheet).
GEN III Microplates, having 96 wells, were Bacterial treatments were tested individually
filled with bacterial suspension, disposing 100 and in combination with half of the chemical
µl in each well. These special plates are dose. This was carried out in order to see if
preloaded with 71 different carbon sources and there is any possibility in reducing the dose of
23 chemicals, which allow several biochemical pesticides for Alternaria spp. prevention.
reactions a single test. Inoculated plates were Samples were incubated at 25°C, optimal for
maintained at 33°C. Within 20 to 48 hours of conidia germination (Troncoso-Rojas and
incubation, plates were spectrophotometrically Tiznado-Hernández, 2014). The observations
analysed at 590 nm and 750 nm wavelengths, at were made after 72 hours of incubations, using
the Biolog Microstation Reader. The readings a light microscope. The test was performed in
were processed with the MicroLog3 software, triplicates and repeated twice.
which identifies the microbial strains based on
their phenotypic pattern. Plant growth promotion
This test was performed on tomato seedlings.
Three experimental variants were studied, two
Enzymatic assays
biological treatments, 75.1s and Cp.b4 respec-
Bacterial ability to produce hydrolytic enzymes
tively, comparing to an untreated control.
was tested on different growth substrates
Bacterial suspensions of 108 cfu/ml were first
(Sicuia et al, 2015). Bacterial ability to solu-
applied as seed treatment, by immersion, for 20
bilize tricalcium phosphate was tested on
minutes. The second treatment was applied to
Pikovskaya agar medium. Chitinase production
the soil, after transplantation, with 2 ml of
was determined on Roberts and Selitrennikoff
bacterial suspension near each plant. Seedlings
(1988) medium with colloidal chitin from crab
were transferred in 10 cm diameter pots. Plants
shell. Cellulase activity was tested on carboxy-
were grown in controlled conditions using a
methil cellulose containing medium, and
Sanyo MLR351H growth chamber, in 16 hours
revealed with 0.1% Congo red. Lipase activity
photoperiods, at 24ºC/light, 18ºC/dark, 14000
was evaluated on Tween 80 supplemented
lx and 70% RH.
medium (mTMB), and protease production was
Biometric observations were made six weeks
tested on slim milk agar.
after transplantation. From each experimental
variant, eight plants were analysed. Root length
Antifungal assay was determined through direct measurement.
The biocontrol activity of the studied/ Fresh weight of total aerial growth and roots
mentioned bacterial strains was evaluated in were separately weighted. Dry weight was
vitro, using the dual culture technique, on determined after 3 hours of soaking at 105°C.
Potato-Dextrose-Agar medium. The antifungal
662
Assimilatory pigments were quantified through Enzymatic characterisation
specific analytical methods. One gram of The bacterial strains were able to produce
freshly harvested leaves, from different plants chitinase, carboxymethil cellulase (CMC-ase),
of each experimental variant, were used in caseinase, and lipase enzymes (Figure 2). Some
order to quantify chlorophyll a (chl a), chlo- of these enzymes are involved in the biocontrol
rophyll b (chl b), xanthophyll and carotenoids. strategy, degrading insect and fungal cell wall
Pigments were extracted in acetone (99.99%) at structure (Veliz et al., 2017).
4°C, overnight. The chlorophyll, xanthophyll Chitinase activity was detected on colloidal
and carotenoids content were quantified accor- chitin media containing bromocresol purple as
ding to Lichtenthaler and Welburn (1987). pH indicator dye (pH 4.7). Therefore, the
Thereafter, the filtered extracts were spectro- breakdown of chitin into N-acetyl glucosamine
photometrically analysed at 662 nm, 645 nm is modifying the pH from acid (4.7) towards
and 470 nm, respectively. The absorbance alkaline, thus changing the colour of the pH
measurements were used for assimilatory indicator dye, from yellow to purple. As a
pigments quantification. Their concentration result of chitinase activity, a purple zone was
was calculated, in mg/g of fresh weight, using generated around the bacterial colonies after 5
Lichtenthaler’s equations. days of incubation on the specified medium
(Figure 2 a). Regarding CMC and casein
RESULTS AND DISCUSSIONS hydrolysis, the enzyme producing strains were
surrounded by a clear hallo due to the cellulase
Bacterial identification and protease activity. The CMC-ase production
The identification was made based on bacterial was revealed by flooding the plates with 0.1%
phenotypic fingerprint obtained in the GEN III Congo red solution for 15 minutes, followed by
Microplates. These plates started out colour- 1M sodium chloride rinses (Figure 2 b, c). The
less. However, during incubation, an increased hydrolysis of Tween 80 from mTMB medium,
respiration, due to carbon source utilization or due to lipase activity, generated a white
bacterial growth, reduced the tetrazolium redox precipitate around bacterial colonies as a result
dye, forming a purple color in specific wells, of calcium soap production (Figure 2 d).
giving a phenotypic fingerprint for each
bacterial strain (Figure 1). The color intensity
was compared with the negative control for
carbon source utilization assays, and positive
control for chemical sensitivity assays. The
phenotypic pattern was compared to Biolog’s
extensive species library, and the identification
was given at specie level. Both strains were
identified as Bacillus cereus/thuringiensis.
Figure 2. Bacterial hydrolytic activity:

a. chitinase, b. CMC-ase, c. protease, d. lipase
The bacterial ability to solubilize tricalcium

phosphate from Pikovskaya agar medium
indicates the bacterial potential in increasing
plant growth through nutrient availability.
Figure 1. Phenotypic profile of Bacillus cereus/ Secondarily, an improved phosphorus uptake is
thuringiensis Cp.b4 on Biolog GEN III microplate under
B type protocol (after 20 hours of incubation at 33°C)
also improving plant resistance to the
phytopathogenic attacks.
663
Antifungal activity Fungal cell lysis associated with protoplasmatic
The antifungal potential of the studied bacterial content likings were seen at the microbial
strains was revealed, by dual culture assay, interaction zone between Botrytis cinerea and
against (new isolate from tomato plant), both bacterial strains (figure 3 a, c). Hyphal
Botrytis cinerea (new isolate from tomato growth deformations were also seen in
fruit), Fusarium oxysporum f. sp. radicis Fusarium oxysporum and Rhizoctonia solani at
lycopersici ZUM 2407, Pythium debaryanum the interaction zone with the biocontrol bacteria
DSM 62946 and Rhizoctonia solani DSM (figure 3 b). The bacterial strains induced
63002 (Table 1). swelling of the fungal cells of Fusarium
We consider that the antifungal potential of oxysporum (figure 3 c), and mycelia
these strains is related with the lytic enzyme dehydration in Botrytis cinerea after prolonged
production and direct competition for nutrients incubation (figure 3 d). Similar aspects were
and niche. previously described on plant pathogenic fungi
when Bacillus spp. treatments were used for
Table 1. Antifungal spectrum of the studied biological control (Huang et al., 2012; Boiu-
Bacillus cereus/thuringiensis strains Sicuia et al., 2017a, b).
Microbial strains 75.1s Cp.b4 Regarding the bacterial potential to inhibit
Alternaria sp. +++ +++ conidia germination, we noticed that all tested
Botrytis cinerea ++ +++ treatments reduced the number of germinated
Fusarium oxysporum f.
++ + conidia of Alternaria sp. (Table 2). The most
sp. radicis lycopersici
Pythium debaryanum + + efficient treatments were the chemical control
Rhizoctonia solani +++ +++ and the mixed applications of biocontrol
Where: + + + = strong inhibition of the fungal growth; + + = moderate bacteria and chemical pesticide in half dose.
inhibition of the fungal growth; + = slight inhibition of the fungal Among bacterial treatments, 75.1s had a better
growth; – = no inhibition of the fungi
biocontrol effect than Cp.b4 strain.
Figure 3. Fungal growth deformations and lysis due to biocontrol bacteria:

A, D = Botrytis cinerea, B = Rhizoctonia solani, C = Furarium oxysporum f.sp. radicis lycopersici
l = cell lysis and cytoplasm likings, c = mycelial curling, s = fungal cell swelling, d = mycelia dehydration
Table 2. Inhibition of Alternaria sp. conidia germination

Germinated Inhibition of Inhibition of germinating
Experimental variant conidia germinated conidia filaments
(%)
Alternaria sp. - Untreated control 32.14 - -
Chemical control
2.7 91.6 86.7
0.2% Mycoguard
Cp.b4 biocontrol treatment 12.5 61.1 98.7
Mixed treatment Cp.b4 &
2.33 92.8 99.1
0.1% Mycoguard
75.1s biocontrol treatment 6.46 79.2 98.7
Mixed treatment 75.1s &
3.33 89.6 99.1
0.1% Mycoguard
The germination filaments were measured filaments length was quantified compared to
using the APS Assess 2.0 software, and the conidia dimensions. Concerning this, the
664
biocontrol and mixt treatments had a better A positive influence of the bacterial treatments
influence to inhibit the growth of germination on tomato seedlings was also seen when the
filaments in Alternaria sp. Although the assimilatory pigments were quantified (Table
chemical control reduced the number of 4).
germinated conidia, the germination filaments
were longer than the other treatments (Table 2). Table 4. Assimilatory pigments in the leaves
of tomato seedlings
Plant growth promoting activity Untreated
Bacterial inoculated
Tomato seedlings were grown in controlled Biometric plants
control
parameters Cp.b4 75.1s
conditions. Six weeks after transplantation,
mg/ g of fresh weight
several biometric observations were made Chlorophyll a 2.435 3.041 3.052
(Table 3). Root length, in cm, was measured, Chlorophyll b 0.949 1.231 1.168
fresh and dry weight of total aerial growth and Xanthophyll and
0.487 0.580 0.610
roots were separately determined as grams of Carotenoids
plant tissue per single plant.
Growth chamber tests on tomato seedlings have Regarding the chlorophyll content, bacterial
shown that biological treatments increase plant treatments slightly increased chl a and chl b
vigour. Plant growth promotion was observed content compared to the untreated control, with
both on aerial parts and root systems when no significant differences among the applied
seedlings were treated with beneficial bacteria. treatments. Moreover, for xanthophyll and
carotenoids, the pigments content was a little
Table 3. Biometric parameters of bacterial higher in treated plants. These results showed
inoculated tomato seedlings an increased photosynthetic capacity in
Bacterial bacterial inoculated seedlings.
Untreated inoculated
Biometric parameters
control plants CONCLUSIONS
Cp.b4 75.1s
Root length (cm) 13.6 17.9 15.7
Aerial fresh weight Two bacterial strains of Bacillus cereus/
5.7675 9.1043 7.2463 thuringiensis, Cp.b4 and 75.1 s, were analysed
(g/plant)
Aerial dry weight (g/plant) 0.5391 0.9630 0.7778 in this study. Both strains demonstrated in vitro
Root fresh weight biocontrol activity and plant growth promotion
0.6345 1.0500 0.7952
(g/plant) properties on tomato seedlings.
Root dry weight (g/plant) 0.0380 0.0819 0.0639
Both strains produced chitinase, carboxymethil
cellulase, caseinase, and lipase enzymes. Due
An increase in root length and weight was
to their high metabolic activity this biocontrol
achieved for the bacterial inoculated plants
strains expressed a wide antimicrobial action
(Figure 4), and a positive correlation of the root
against Alternaria sp. (new isolate from tomato
system with the shoots was revealed. Between
plant), Botrytis cinerea (new isolate from
the two tested bacterial strains, Cp.b4 was more
tomato fruit), Fusarium oxysporum f. sp.
efficient than 75.1s in plant growth promotion
radicis lycopersici ZUM 2407, Pythium
(Table 3).
debaryanum DSM 62946 and Rhizoctonia
solani DSM 63002. The biocontrol strains also
inhibited conidia germination in Alternaria sp.,
showing a high potential in prevent early blight
infections. Biological treatments with bacterial
suspension reduced the number of germinated
conidia by 4.98 fold (when using 75.1s strain)
and by 2.57 fold (when using Cp.b4 strain) than
the untreated control. Mixed treatments of
bacteria suspension and low dose (0.1%) of
Figure 4. Tomato roots: a) untreated control; chemical fungicide strongly inhibit conidia
b) 75.1s treated seedlings; c) Cp.b4 treated seedlings germination, being comparable with the
665
chemical control, Mycoguard 500SC, applied at solani as influenced by different delivery methods of
the recommended dose of 0.2%. Pseudomonas gladioli B25. Acta Horticulturae, 808:
327-332.
Regarding plant growth promotion activity, the Kennelly M., O'Mara J., Rivard C., Miller G.L., Smith
mentioned strains, applied as seed and soil D., 2012. Introduction to abiotic disorders in plants.
treatment, increased tomato root length, The Plant Health Instructor. DOI: 10.1094/PHI-I-
biomass fresh and dry weight, and plant 2012-10-29-01
photosynthetic capacity. The bacteria also Lichtenhaller H.K., Wellburn A.R., 1987.
Determinations of total carotenoids and chlorophylls a
revealed the ability to solubilize tricalcium and b of leaf extracts in different solvents.
phosphate from Pikovskaya agar medium Biochemical Society Transactions 591- 592.
indicating an increasing potential in improving Roberts W.A., Selitrenikoff C.P., 1988. Plant and
nutrient availability for the plants. bacterial chitinase differ in antifungal activity. Journal
of General Microbiology, 134: 169-176.
Sicuia O.A., Grosu I., Constantinescu F., Voaideş C.,
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Agamy R., Alamri S., Moustafa M.F.M., Hashem M., AgroLife Scientific Journal, 4 (2): 124-131.
2013. Management of tomato leaf spot caused by Thomma B.P.H.G., 2003. Alternaria spp.: from general
Alternaria tenuissima (Kunze ex Pers.) Wiltshire saprophyte to specific parasite. Molecular Plant
using salicylic acid and agrileen. International Journal Pathology, 4 (4): 225-236.
of Agriculture & Biology, 15: 266-272. Troncoso-Rojas R., Tiznado-Hernández M.E., 2014.
Biolog, Inc., US Patent # 5,627,045. Alternaria alternata (black rot, black spot). Chapter 5
Boiu-Sicuia O.A., Constantinescu F., Cornea C.P., 2017. in Bautista-Banos S. (ed.), Postharvest decay. Control
Selection and characterization of new endophytic strategies. Elsevier, 147-187.
bacterial strains isolated from potato tuber useful in Veliz E.A., Martínez-Hidalgo P., Hirsch A.M., 2017.
biocontrol strategies. Scientific Bulletin. Series F. Chitinase-producing bacteria and their role in
Biotechnologies, XXI: 23-28. biocontrol. AIMS Microbiology, 3 (3): 689-705.
Boiu-Sicuia O.A., Constantinescu F., Diaconu A., Wang Y., Bao Y., Shen D., Feng W., Yu T., Zhang J.,
Drăghici R., 2017. Research approaches regarding Zheng X.D., 2008. Biocontrol of Alternaria alternata
biological control of Fusarium sp. stem rot of sweet on cherry tomato fruit by use of marine yeast
potato produced on sandy soils. Muzeul Olteniei Rhodosporidium paludigenum Fell & Tallman.
Craiova. Oltenia. Studii şi comunicări. Ştiinţele International Journal of Food Microbiology, 123 (3):
Naturii, 33 (2): 171-178. 234-239.
Chaerani R., Voorrips R.E., 2006. Tomato early blight Woudenberg J.H.C., Seidl M.F., Groenewald J.Z., de
(Alternaria solani): the pathogen, genetics, and Vries M., Stielow J. B., Thomma B.P.H.J., Crous
breeding for resistance. Journal of General Plant P.W., 2015. Alternaria section Alternaria: Species,
Pathology, 72: 335-347. formae speciales or pathotypes? Studies in Mycology,
Huang X., Zhang N., Yong X., Yang X., Shen Q., 2012. 82:1-21.
Biocontrol of Rhizoctonia solani damping-off disease Zahoor A., Jia W., Lulu C., Wubei D., 2017. Isolated
in cucumber with Bacillus pumilus SQR-N43. Bacillus subtilis strain 330-2 and its antagonistic
Microbiological Research, 167: 135-143. genes identified by the removing PCR. Scientific
Jagadeesh K.S., Jagadeesh D.R., 2009. Biological Reports, 7: 1777.
control of early blight of tomato caused by Alternaria
666
ANTIMICROBIAL POTENTIAL OF ROMANIAN SPONTANEOUS FLORA -

A MINIREVIEW
Tomina PURCARU, Camelia DIGUŢĂ, Florentina MATEI

Faculty of Biotechnologies, 59 Mărăşti Blvd, District 1, Bucharest, Romania
Abstract
In the past decades, clinical microbiologists, practitioners and professionals in food safety, are facing new challenges
related to new born microbial pathogens as well as to the phenomenon of the antibiotic and biocide resistance
developed by the pathogens. Meanwhile, in Romania has been noticed an increase in scientific publications dealing
with the potential of Romanian aromatic and medicinal plants and their therapeutic use. The paper proposes a mini-
review on scientifically proved antimicrobial activity of aerial and underground parts of some spontaneous plant from
Romanian flora. The review approaches annual and perennial plants, from herbaceous species to bushes and trees. In
our search we have identified a total of 64 species from autochthonous flora involved in studies on antimicrobial
activity, belonging to 21 botanical families. Among these species, 28.1% are annual plants, 46.9% are herbaceous
perennial plants and the rest (25%) are woody perennial species (bushes and trees). Almost 50% of the active species
belongs to Asteraceae and Lamiaceae botanical families. For 89% of the species have been reported antibacterial
activity, while only 57.8% of the species have proven antifungal activity.
Key words: Romanian spontaneous flora, antibacterial, antifungal, phytopharmaceutical use.
INTRODUCTION different mechanisms, more or less elucidated,

the microorganisms became over and over
In the past decades, clinical microbiologists, resistance to the new antibiotics and biocids.
practitioners and professionals in food safety, Meanwhile, with the growing consumer
are facing new challenges related to new born demand for natural preservatives to replace
microbial pathogens as well as to the chemical compounds, plant antimicrobial
phenomenon of the antibiotic and biocide compounds must be thoroughly investigated for
resistance developed by the pathogens, of their potential to serve as natural preservatives
which the most studied is methicillin-resistant (Hintz et al, 2015); on this side, very recently,
Staphyloccous aureus MRSA (Lee et al., 2015). an exhaustive overview on natural food
The antibiotic resistance crisis has been preservatives with antimicrobial properties has
attributed to the overuse and misuse of the been reported by Pisoschi et al. (2018).In this
medications, as well as the lack of new drug context, the “return to the origins”, meaning the
development by the pharmaceutical industry use of the spontaneous flora as antimicrobial
due to reduced economic incentives and tool may be a solution of the antibiotic “crisis”
challenging regulatory requirements (Ventola, and the replacement of chemical preservatives.
2015). In ancient times, microbial infections A lot of studies have been published in the past
were treated empirically, by the use of different two decades and it has been proven that
natural solutions, including the use of different spontaneous flora of each continent,
plants prepared under different formulations. hemisphere, country or region has an immense
Starting with the discovery of penicillin by Sir potential in obtaining different products with
Alexander Fleming in 1928, antibiotics have antimicrobial and anti-inflammatory impact.
transformed modern medicine. Over the past The repository of the whole information is
decades, starting with 1950, it has been proven quite difficult because of the huge volume of
that bacteria have developed resistance to data, but there are some reports related to
different antibiotics; as a consequence, new different countries or regions. For example, in
synthesis substances have been developed to Balkan regions (Southeast Europe), where
treat the infections (Ventola, 2015); because of Romania belongs, an ethnobotanical analysis
667
showed that 128 plant species (105 wild, 22 proposes to present an overview of Romanian
cultivated and 1 wild/cultivated) are used in the spontaneous flora proved by experimental
treatment of wounds. Their application is approach to have antimicrobial effect.
external, in the form of infusions, decoctions,
tinctures, syrups, oils, ointments, and balms, or MATERIALS AND METHODS
direct to the skin. Among those plants recorded,
the most commonly used in Balkans are Online information research was conducted by
Plantago major, Hypericum perforatum, the use of different database collections and on-
Plantago lanceolata, Achillea millefolium, searching engines (Google Academic, Web of
Calendula officinalis, Sambucus nigra, Knowledge, PubMed, ScienceDirect and
Tussilago farfara and Prunus domestica (Jaric Embase, InTech and Hindawi databases). The
et al., 2018). As in all the other countries or information has been structured in relation to
regions, on the ancient lands of the actual the plants type (annual/biennial and perennial
Romania, it was a vivant interest in the use of plants, from herbaceous species to bushes and
different plants to treat human, animal or even trees).Where information was available, the
plants’ infections. Jaric et al. (2018) make district or the county of the plant origin have
references to different studies and report that in been specified.
Romania, out of more than 3600 species of
plants, over 700 are medicinal plants. RESULTS AND DISCUSSIONS
Nowadays the interest has been resuscitated
and more complex studies have been performed The database screening has led to a collection
related to the chemical composition of the of over sixty scientific publications, dated from
popular plants, as well as on testing their 2007 to 2018. The authors have reported the
antimicrobial effect by the use of a use of different types of extracts (aqueous,
standardized methodological approach. On our alcoholic or PEG extracts), as well as the use
knowledge, there is no comprehensive review essential oils of different plants in testing the
on the published studies developed on antimicrobial activity. Different methods have
Romanian level in relation to our local flora been used to test the antimicrobial activity of
and its antimicrobial properties. However, different plants products. The most usual
Amarioarei et al. (2016) has published a method is the agar diffusion method described
scientometric analysis performed on data by different authors or clinical standards in
collected from Scopus, during 2000-2015; the USA or Europe (Brown, 1978; Das et al.,
review contains information on number of 2010). The findings are described in the
papers, citations, affiliation and number of following, grouped in annual or perennial
authors dealing with the potential of Romanian plants, herbaceous or woody groups.
aromatic and medicinal plants and their
therapeutic use. The authors have reported an 1) Annual / Biennial plants
increasing trend for such publications in The reported antimicrobial activity of
Romania starting with 2007 which we could annual/biennial plants from Romanian flora are
rely with the variety of funding available synthetized in Table 1 and their appurtenance
through the national and regional funding to botanical family is clearly specified.
programs. Also, there is some information There are different studies targeting the
provided from some Romanian geographical botanical family Lamiaceae, which includes
regions on plants having different most of the aromatic plants like Origanum
phytopharmaceutical effects; for example, in vulgare L., Melissa officinalis L. or Ocimum
Banat region were identified about 140 plant basilicum L.; ethanolic extracts of aerial parts
species with antioxidant potential (Antal, of these plants have been proven to have some
2010). Generally, antioxidant activity is due to antibacterial activity on Listeria
polyphenol content, especially flavonoids monocytogenes, Stahpylococcus aureus
compounds, and it is assumed that same (Benedec et al., 2015), as well as antifungal
compounds are partially responsible for the activity on Candida albicans (Tuchila et al.,
antimicrobial activity. The present paper 2008; Benedec et al., 2015).
668
Table 1. Romanian annual herbaceous plants with antimicrobial activity
Plant species Botanical Plant part Antimicrobial activity Reference
family Bacteria Fungi
Origanum vulgare L. Lamiaceae hb; fl; fs Listeria monocytogenes Candida albicans Benedec et al., 2015
Stahpylococcus aureus Dobre et al., 2011
E. coli Sandru et al., 2015
Salmonella enteridis
Melissa officinalis L. Lamiaceae hb; fl; fs Listeria monocytogenes Candida albicans Benedec et al., 2015
Stahpylococcus aureus Hancianu et al., 2008
Ocimum basilicum L. Lamiaceae hb; fl; fs Listeria monocytogenes Candida albicans Benedec et al., 2015
Staphylococcus aureus Stefan et al., 2011
E. coli Tuchila et al., 2008
Streptococcus cricetus Vlase et al., 2014
Thymus vulgaris L. Lamiaceae hb; fl; fs Staphylococcus aureus Candida albicans Boruga et al., 2014.
Klebsiella pneumoniae Aspergillus niger Dobre et al., 2011
Salmonella typhimurium Grigore Armatu et al.,
E. coli 2012
Enterococcus faecalis Varga et al., 2015
Salmonella enteridis
Pseudomonas aeruginosa
Listeria innocua
Streptococcus pyogenes
Thymus pulegioides hb E. coli Candida albicans Pavel et al., 2010
Enterobacter cloacae Varga et al., 2015
Proteus mirabilis
Bacillus subtilis
Micrococcus flavus
Lamiaceae
Staphylococcus aureus
Streptococcus faecalis
Listeria innocua
Thymus glabrescens Lamiaceae hb Salmonella typhimurium Candida albicans Pavel et al., 2010
Pseudomonas aeruginosa Varga et al., 2015
Proteus mirabilis
Listeria innocua
Satureja hortesis Lamiaceae hb Streptococcus cricetus Botrityis cinerea Sesan et al., 2015
Staphylococcus aureus Candida albicans Tuchila et al., 2008
Anethum graveolens Apiaceae hb; fl; fs; Shigella flexneri Jianu et al., 2012
sm Klebsiella pneumoniae
Salmonella typhimurium
E. coli
Tropaeolum majus Tropaeolaceae hb Pseudomonas aeruginosa Candida albicans Butnariu et Bostan, 2011
Salmonella sp.
Bacillus sp.
Veronica officinalis Plantaginaceae hb Listeria monocytogenes - Mocan et al., 2015a
Listeria ivanovii Mocan et al., 2015b
Veronica teucrium Plantaginaceae hb Staphylococcus aureus - Mocan et al., 2015a
Bacillus cereus Mocan et al., 2015b
Enterococcus faecalis
Peptostreptococcus anaerobius
Veronica orchidea Plantaginaceae hb Listeria monocytogenes - Mocan et al., 2015a
Listeria ivanovii Mocan et al., 2015b
Peptostreptococcus anaerobius
Veronica persica Poiret Plantaginaceae hb; fl; fs Aspergillus niger Fierascu et al., 2018
Penicillium hirsutum
Arctium lappa Asteraceae rx E. coli Aspergillus niger Fierascu et al., 2018
Salmonella abony Penicillium hirsutum Ionescu et al., 2013
fl Staphylococcus aureus Pirvu et al., 2017
Staphylococcus epidermidis
Xanthium strumarium Asteraceae hb - Phytophthora infestans Rodino et al., 2013
Cnicus benedictus Asteraceae fs Salmonella typhimurium - Szabo et al., 2009
Salmonella enteritidis,
Shigella sonnei
Proteus vulgaris,
E. coli,
Pseudomonas aeruginosa,
Calendula officinalis Asteraceae hb Klebsiella penumoniae Candida albicans Jianu et al, 2016
S. aureus
E. coli
Tagetes patula Asteraceae fs Pythium sp. Rodino et al., 2015 a
Botrytis cinerea Sesean et al., 2015
Legend: hb: herba (flowering aerial parts); fl: folium (leaves); fs: flos (flowers); nd: needles; fr: fructus (fruits); cx: cortex (bark); sm: semen (grains); rx: radix (roots); rh:
rhizoma (rhizome); st: stipites (branches).
669
Similar results on essential oil of Melissa inhibit the growth of Pseudomonas aeruginosa,
officinalis L. have been reported by Hancianu Listeria innocua and Streptococcus pyogenes.
et al. (2008). Regarding Origanum vulgare, In the same family (Lamiaceae), Satureja
Sandru et al. (2015) have proven that essential hortensis harvested in Southern Romania has
oils made of the aerial parts have strong been reported to have antifungal effect on
inhibitory effect on E.coli, same as other Botrytis cinerea (Sesan et al., 2015). Aqueous
different species of Ocimum used as essential extracts of Satureja hortensis from Banat
oils (Stefan et al., 2011). Ocimum sp. spectrum county have inhibited Streptococcus cricetus,
is completed by Streptococcus cricetus which while in alcoholic extract inhibited
is inhibited by the alcoholic extract (Tuchila et Staphylococcus aureus and Candida albicans
al., 2008). (Tuchila et al., 2008).
In the same family, essential oils of Thymus The list of aromatic plants with antimicrobial
vulgaris aerial parts, harvested in Mehedinţi activity is also completed by the dill. Essential
County, have moderate to strong inhibition on oils from inflorescences, stems, immature and
Staphylococcus aureus, Klebsiella pneumoniae, mature seeds of Anethum graveolens L. grown
Salmonella typhimurium, E. coli, Enterococcus in Western Romania (Timis county) were
faecalis and Candida albicans (Boruga et al, isolated by steam distillation and tested on
2014); the authors correlate this activity with different bacteria. Significant antimicrobial
the presence of phenolic compounds (thymol) activity was recorded against Shigella flexneri,
and terpene hydrocarbons (γ-terpinene). Some Klebsiella pneumoniae, Salmonella
other authors reported higher antibacterial typhimurium and E. coli, while no inhibitory
activity of thyme extracts originated in effects were observed against Streptococcus
Southern Romania (Dobre et al., 2011), as well pyogenes and Staphylococcus aureus, results
as anti-fungal effect (Grigore Armatu et al., which is partially in contradiction with other
2012). reported results (Jianu et al., 2012).
Other Thymus species have been investigated. From a plant mainly cultivated as ornamental
Aerial parts of Thymus pulegioides collected at plant, Tropaeolum majus, Butanriu and Bistan
the flowering stage from two areas of the (2011) have extracted essential oils starting
Bucegi Mountains at different altitudes (1000 from dehydrated leaves and flowers harvested
and 1800 m above sea level) and aerial parts of in Timis county. The authors assumed that the
T. glabrescens from the district of Gorj have antimicrobial action is determined by the
been used for essential oils extraction (Pavel et phenols and metil-ethers identified in the T.
al., 2010). Escherichia coli, Enterobacter majus extracts, but also by the tymol and
cloacae, Proteus mirabilis, Bacillus subtilis carvacrol present in the volatile oil; the volatile
and Micrococcus flavus were the strains most oils tested presented a wide range of action
susceptible to T. pulegioides essential oil. over both Gram-positive and Gram–negative
T.glabrescens essential oil inhibited the growth species. The most sensitive microorganism to
of Salmonella typhimurium, Pseudomonas the action of the tested natural compounds of T.
aeruginosa and Proteus mirabilis. The authors majus proved to be P. aeruginosa and C.
have related the inhibitory activity tothe albicans, followed by Salmonella sp. and
presence of monoterpenoid alcohols in this Bacillus sp, while the most resistant is the E.
sample, especially of geraniol (55.5%), which coli stem.
manifests an antiseptic activity comparable to A plant considered annual, but sometime being
that of thymol, often against Pseudomonas. All an over winter specie, is Veronica persica
the tested samples showed antifungal effects by Poiret. Crude hydroalcoholic extracts of its
inhibiting the growth of Candida albicans. aerial parts originated in Pitesti hills showed
The Thymus sp. spectrum of antibacterial important inhibitory effect on two pathogenic
activity is completed with data reported by fungal species, Aspergillus niger and
Varga et al. (2015). Essential oils of four Penicillium hirsutum (Fierascu et al., 2018).
different Thymus species (T. vulgaris, T. This results have completed the image of
serpyllum, T. pulegioides, and T. glabrescens) antimicrobial effect of Veronica sp. described
harvested in Mures county have been proven to by Mocan et al. (2015), which have proven that
670
V. officinalis, V. teucrium and V. orchidea have typhimurium, Salmonella enteritidis to Shigella
inhibitory effect on Staphylococcus aureus, sonnei, Staphylococcus aureus, Streptococcus
Listeria monocytogenes and Listeria ivanovii. pyogenes, Proteus vulgaris, Escherichia coli,
Other species of Veronica genus have been Pseudomonas aeruginosa and Enterococcus
studied by Mocan et al. (2015 a, b). faecalis (Szabo et al., 2009).
Hydroalcoholic extracts of aerial parts From Asteraceae family have been taken into
harvested in Cluj county shows that in the case account also a species growing as weed,
of V. officinalis, the most sensitive bacterial Xanthium strumarium. Ethanolic extracts of
strains were Listeria monocytogenes and aerial parts of the plant have been demonstrated
Listeria ivanovii; regarding V. teucrium to have inhibitory effect on the growth of a
antibacterial activity, the strains of phytopathogenic fungi, Phytophthora infestans,
Staphylococcus aureus, Bacillus cereus, the causative agent of late blight in tomatoes
Enterococcus faecalis have been the most and potatoes (Rodino et al., 2013). Further,
sensitive. Referring to the V. orchidea extract, Rodino et al. (2015 a) have tested another
the most sensitive strains were Listeria Asteraceae representative, Tagetes patula
monocytogenes and Listeria ivanovii. Also, the (marigold) on the phytopathogenic fungi
extracts of V. teucrium and V. orchidea have Pythium sp., which can cause serious diseases
been proven to have antibacterial activity, on such as damping off, seed rot, root rot and soft
Peptostreptococcus anaerobius, an anaerobic rot in wheat, maize, soybean, peppers, bean,
Gram-positive bacteria responsible for clinical cucumber, tomato. Ethanolic extracts of
infections. The authors come with the marigold flowers harvested from Southern
assumption that the activity of Veronica Romanian from non-polluted sites, exhibited
ethanolic extracts against Gram-positive moderate to high inhibition on the fungal
bacteria like L. monocytogenes, L. ivanovii and specie. Other authors (Sesan et al., 2015)
S. aureus could be attributed at least in part to reported Tagetes sp. extracts as having good
their high β-sitosterol content but also to the antifungal activity on Botrytis cinerea.
presence of campesterol and stigmasterol and Another annual member of Asteraceae family,
may be might be influenced also by the Calendula officinalis, rarely studied for its
presence of hispidulin. Relatively recently, antimicrobial activity, as essential oils of aerial
have been given special attention to the parts has inhibited the growth of Klebsiella
biennial Arctium lappa. Crude roots penumoniae, S. aureus, E. coli and of the
hydroalcoholic extract proved inhibitory effect fungus Candida albicans (Jianu et al., 2016).
on Escherichia coli, Salmonella abony (Ionescu There are some other annual plants studied for
et al., 2013), as well as on fungi as Aspergillus their antimicrobial activity, activity which have
niger and Penicillium hirsutum (Fierascu et al., been proven to be weak on Romanian extracts,
2018). Authors have attributed the even some other reports are opposite. An
antimicrobial properties to the phenolic acids example is Agrimoniae herba ethanolic extract
content (such as chlorogenic acid, rutin, which has only a weak inhibitory effect on
quercitrin, luteolin, p-coumaric acid, caffeic Pseudomonas aeuroginosa (Pirvu et al., 2016).
acid and quercetin). In 2017, results obtained
by Pirvu et al. suggest the potential uses of 2) Perennial plants
Arctii folium whole (70%, v/v) ethanol extract The reported antimicrobial activity of perrenial
in restoring the activity of the antibiotics plants from Romanian flora are synthetized in
affected by microbial resistance, as well as Table 2 (herbaceous plant) and Table 3 (shrubs
inhibitory effect on Stapyloccous epidermidis. and trees), including their appurtenance to
A thistle-like plant from Asteraceae family, botanical family and the plants’ part tested for
Cnicus benedictus in different extracts of the inhibitory activity.
immature capitulum harvested in North- Herbaceous plants
Western Romania during prebloom period, Different species from Asteraceae family have
have been proven to have a very large been proven to have inhibitory activity on
antibacterial spectrum, from Salmonella pathogenic microorganisms. Achillea sp.
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Table 2. Romanian perennial herbaceous plants with antimicrobial activity
Plant species Botanical Plant part Antimicrobial activity Reference
family Bacteria Fungi
Achillea distans Asteraceae fs Listeria monocytogenes Benedec et al., 2013
Stahpylococcus aureus
Achillea shurii Asteraceae fs Listeria monocytogenes Benedec et al., 2016
Achillea millefolium Asteraceae hb Klebsiella pneumoniae Candida albicans Jianu et al., 2016
Achillea collina Asteraceae fs E. coli, Shigella flexneri Jianu et al., 2015
Klebsiella pneumoniae
Artemisia spp. Asteraceae fs Sclerotinia sclerotiorum Badea and Delian, 2014
Tanacetum vulgare Asteraceae fs Bacillus cereus Muresan, 2015
Staphylococcus aureus Muresan et al., 2015
Inula helenium Asteraceae rx E. coli, Enterococcus faecalis Candida albicans Diguta et al., 2014
Bacillus cereus Candida parapsilosis
Santolina rosmarinifolia Asteraceae hb; fl; fs Staphylococcus aureus Candida albicans Ioannou et al., 2007
Cynara scolymus Asteraceae fl E. coli, Salmonella abony Ionescu et al., 2013

Listeria innocua, Bacillus cereus Vamanu et al., 2011
Taraxacum officinale Asteraceae fl Escherichia coli Ionescu et al., 2013
Salmonella abony
Eupatorium cannabium Asteraceae fl Escherichia coli Candida albicans Purcaru et al., 2015
Bacillus subtilis
Salvia officinalis Lamiaceae hb; fl; fs Listeria monocytogenes Candida albicans Benedec et al., 2015
Stahpylococcus aureus Ilie et al., 2016
Klebsiella pneumoniae
Rosmarinus officinalis Lamiaceae hb; fl; fs Listeria monocytogenes Candida albicans Benedec et al., 2015
Stahpylococcus aureus Aspergillus flavus
Aspergillus ochraceus
Hyssopus officinalis Lamiaceae hb; fl; fs Listeria monocytogenes Candida albicans Benedec et al., 2015
Stahpylococcus aureus Jianu et al., 2016
P. aeruginosa Mihai and Popa, 2015
Vlase et al., 2014
Mentha piperita Lamiaceae hb - Candida albicans Jianu et al., 2016
Botrytis cinerea Sesan et al., 2015
Mentha smithiana Lamiaceae hb - Candida albicans Jianu et al., 2016
Mentha spicata Lamiaceae hb Listeria monocytogenes Candida albicans Moldovan et al., 2014
Mentha rotundifolia Lamiaceae hb Listeria monocytogenes Candida albicans Moldovan et al., 2014
Ajuga genevensis Lamiaceae hb Staphylococcus aureus Toiu et al., 2016
Listeria monocytogenes,
E. coli, Salmonella typhimurium
Teucrium chamaedrys Lamiaceae hb Staphylococcus aureus Candida albicans Vlase et al., 2014
Hypericum perforatum Hypericaceae hb S. aureus, S. typhimurium Candida albicans Jianu et al., 2016
E. coli
Eryngium campestre Apiaceae hb; fl; fs Staphylococcus aureus Conea et al., 2016
Stahpylococcus epidermidis
Humulus lupulus Cannabaceae fs Bacillus subtilis, E. coli - Arsene et al., 2015
Enterococcus faecalis,
Bacillus cereus, Staphylococcus
aureus, Enterobacter cloacae,
Pseudomonas fluorescens,
Hedera helix Araliaceae fs; fr Stahpylococcus aureus Pop et al., 2017
Listeria monocytogenes
Allium ursinum Amaryllidaceae Bacillus subtilis Aspergillus glaucus Lupoae et al., 2013
Allium sativum Staphylococcus aureus Geotrichum candidum
Streptococcus pyogenes Candida albicans
E.coli Botrytis cinerea Sesan et al., 2015
Helianthemum Cistaceae hb E. coli, Staphylococcus aureus Candida albicans Pirvu et al., 2017a
nummularium Salmonella typhimurium
Salmonella enterritidis
Pseudomonas aerugionsa
Epilobium hirsutum Onagraceae hb Staphylococcus aureus Pirvu et al., 2014
E. coli Pirvu et al., 2015
Chelidonium majus Papaveraceae hb Botrytyis cinerea Parvu et al, 2011
Glycyrrhiza glabra Fabaceae rx Pythium sp. Rodino et al., 2015a
Paeonia officinalis Paeoniaceae E. coli, Pseudomonas aeruginosa Aspergillus niger Soare et al., 2012
Salmonella abony,
Brevibacterium flavum
Sarcina sp., Bacillus cereus
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Is one of the most studied in the family. aeruginosa (Muresan, 2015; Muresan et al.,
Achillea distans Waldst. et Kit. ex Willd., 2015).
found in the Rodna Mountains (a subdivision The Asteraceae list is completed by Inula
of the Eastern Carpathians in Northern helenium; the ethanolic extracts were obtained
Romania), is confirmed as a native species of from the roots of plants harvested in Brasov
the Romanian flora; its flowers hydroalcoholic county (Transilvania); moderate to high
extract showed inhibitory activity on Gram- bacterial inhibition have been shown on
positive bacteria as reported by Benedec et al Escherichia coli, Enterococcus faecalis,
(2013). From the same family, hydroalcoholic Bacillus cereus and Staphylococcus aureus;
extract of Achillea schurii Sch.-Bip., an meanwhile, moderate anti-Candida effects have
endemic species from Romania, has revealed a been proven (Diguta et al., 2014).
remarkable inhibitory effect on Listeria Essential oils of the flower heads and leaves of
monocytogenes, Staphylococcus aureus and Santolina rosmarinifolia L. were obtained
Salmonella typhimurium (Benedec et al., 2016). through hydrodistillation and tested against
Essential oil of inflorescence harvested from Gram-positive and Gram-negative bacteria
Achillea millefolium and its hybrid Achillea strains and the fungus Candida albicans
collina Becker growing wild in Western (Ioannou et al., 2007). The highest inhibitory
Romania inhibited most strongly the growth of potential has been shown on Staphylococcus
E. coli, followed by Shigella flexneri, aureus and Candida albicans.
Klebsiella pneumoniae, Salmonella In the same family (Asteraceae) two other
typhimurium and Staphylococcus aureus. No species, Cynara scolymus and Taraxacum
effects were observed against Clostridium officinale have proven antibacterial activity in
perfringens and Streptococcus pyogenes (Jianu hydroalcoholic leaves extracts against
et al., 2015; Jianu et al., 2016). The authors Escherichia coli and Salmonella abony
assumed this could be the results of the (Ionescu et al., 2013). Meanwhile, Vamanu et
inhibitory effects exhibited by the major al. (2011) has reported that freeze-dried
constituents of the analyzed essential oils, ethanolic extracts of Cynara scolymus
respectively chamazulene, caryophyllene and harvested in Hunedoara county (Transylvania)
β-pinene; also they noticed the presence of have significant inhibitory effect on Listeria
certain minor components, known for their innocua and Bacillus cereus.
strong antimicrobial activity, such as limonene, Purcaru et al. (2015) have tested different dried
α-pinene or 1.8-cineole. leaves Eupatorium cannabium extracts made of
Essentila oils obtained by hydro distillation, a Romanian cultivar from Brasov county. In the
from Artemisia spp growth in different case of the chloroformic extract and hydro-
Romanian areas, as spontaneous flora or as alcoholic extract the inhibitory activity has
cultivated species have been tested against been noticed only in the case of Escherichia
fungal pathogen Sclerotinia sclerotiorum (Lib.) coli and Bacillus cereus, as well as on the
de Bary, from carrots roots stored in the dimorphic yeast Candida albicans. No clear
refrigerator (Badea and Delian, 2014); inhibition has been noticed in the case of
minimum inhibitory concentration (MIC) was Staphylococcus aureus, Enterococcus faecalis
found to be 2400 μL L-1 for A. santonica, A. and Aspergillus niger.
pontica, A. annua, A. austriaca, A. Lamiaceae family is on the top list of plants
dracunculus, A. lerchiana, A. vulgaris and A. tested for their antimicrobial activity. Benedec
vulgaris var. pilosa. et al. (2015) has proven that the rosmarinic acid
Tanacetum vulgare is known mainly for its from Salvia officinalis L. and Rosmarinus
toxicity and insect repelent properties. Essential officinalis L. has strong antibacterial effect on
oils and ethanolic extracts of this plant, Gram positive bacteria, even higher than
harvested in Transylvania (Sibiu and Alba gentamicin; similarly, strong effect has been
county) exhibited moderate inhibition on noticed against Candida albicans, higher than
Staphylococcus aureus and Bacillus subtilis, fluconazole. Meanwhile, essential oils from
but low activity on E. coli and Pseudomonas aerial part of Salvia officinalis originated in
Arad county showed strong inhibitory effect of
673
Staphyloccus aureus and Klebsiella from E. planum and E. campestre from Cluj
pneumoniae (Ilie et al., 2016). The county and E. maritimum from Constanta
antimicrobial activity recorded have been county have proven to have moderate
attributed mainly to the major components of S. antibacterial activity on Stahpylococcus aureus
officinalis essential oils, i.e., camphor, alpha- and Stahpylococcus epidermidis and high
thujone and alpha-humulene, recognized for inhibitory activity on Pseudomonas
their biological activities. Also, essential oils aeruginosa, especially in the case of E.
and terpens extracted from Rosamarinus campestre (Conea et al., 2016); authors
officinalis have been proven to have antifugal assumed that the activity of Eryngium tinctures
effects, both on growth and sporulation of probably results from the synergistic effect of
Aspergillus flavus and Aspergillus ochraceus; triterpene saponins, polyphenols, sterols, pectin
lower effect have been registered on and other active compounds.
Aspergillus niger (Mihai and Popa, 2015). Humulus lupulus (common hop) from Southern
Remaining in the same Lamiaceae family, in Romania, an herbaceous climbing plant, as
Romania has been reported for the first time hydroalcoholic extracts of female
antimicrobial activity of essential oils of inflorescences, has been proven to have
Mentha smithiana (Jianu et al., 2016). Aside antagonistic effect on both Gram-positive and
Mentha piperita, their essential oils inhibited negative bacteria (Aresene et al., 2015). In the
mainly the Gram-positive bacteria, as well as case of hope the substances associated with
the fungus Candida albicans. Similar results antibacterial activity are humulone, lupulone
have been obtained by the same authors in the and xanthohumol (Cermak et al., 2017).
case of essential oils of Hypericum perforatum. Another climbing plant, Hedera helix harvested
Also. Mentha sp. has been reported to have in Cluj county as leaves, flower and immature
significant antifungal activity on Botrytis fruits has been tested by Pop et al. (2017). They
cinerea (Şesan et al., 2015). Other species of have arrived to the conclusion that the
Mentha genus have been reported for anti- immature fruits extract showed a significant
Candida activity by Moldovan et al. (2014), activityagainst S. aureus, followed by the
respectively extracts of M. spicata subsp. flower extract with a good growth inhibitory
crispata and M. x rotundifolia. Same research effect against the same bacterial strain. Both
group reported that other Mentha sp. extracts immature fruits and flowers extracts possess
have strong inhibitory activity on Listeria appropriate antibacterialcapacity against L.
monocytogenes. monocytogenes.
Aerial part of Ajuga genevensis harvested from Among perennial bulbous of Romanian wild
wild populations from Cluj county at full flora has been tested Allium ursinum in
flowering stage, in alcoholic extracts showed hydroalcoholic or acetic acid extracts obtained
high inhibitory activity against Staphylococcus from different parts (leaves, roots, bulbs). The
aureus, followed by Pseudomonas aeruginosa, extracts inhibited the growth of different
Listeria monocytogenes, Escherichia coli and altering or pathogen fungi like Aspergillus
Salmonella typhimurium (Toiu et al., 2016). glaucus, Geotrichum candidum and Candida
Lamiaceae family list of plants with albicans, as well as on different Gram-positive
animicorbial activity is completed by the and Gram-negative bacteria (Lupoae et al.,
ornamental Teucrium chamaedrys. Aerial parts 2013). The authors recommend their use in the
harvested during summer in Sibiu county food industry as additive. Şesan et al. (2015)
(Transilvania) have been prepared as ethanolic have also demonstrated that Allium sativum
extract; the extract showed stronger extracts have inhibitory effects on Botrytis
antibacterial activity against S. aureus than cinerea (grey mould)affecting cultures of Ribes
gentamicin used as reference antibiotic, as well nigrum.
as antifungal activity against Candida albicans, The hydroalcoholic extract of Chelidonium
higher than fluconazole(Vlase et al., 2014). majus (Papaveraceae) obtained from powder of
From Apiaceae family, different species of dried aerial plant organs collected from a
Erymgium have been tested for their private homegarden in Cluj county had
antimicrobial activity. Tincture of aerial plants antifungal effect against B. cinerea (Pârvu et
674
al., 2011). Another phytopathogenic fungi, harvested in July from Romanian Carpathian
Pythium sp., have been proven to be inhibited Mountains. These extracts show certain
by ethanolic extracts of Glycyrrhiza glabra antimicrobial activity on E. coli, as well as
(Fabaceae) roots harvested in Southern weak to moderate activity on S. aureus, S.
Romania from non-polluted sites (Rodino et al., typhimurium and S. enterritidis; the list is
2015a). completed by Pseudomonas aueriginosa and
A novelty may be considered the studies Candida albicans.
conducted by Pîrvu et al. (2017 a) regarding the Among perennial herbaceous plants tested for
antimicrobial activities of extracts from rock antimicrobial activity an ornamental plant was
rose (Helianthemum nummularium Mill.) in the research attention. Red petals of Paeonia
Table 3. Romanian perennial woody plants with antimicrobial activity
Botanical Plant Antimicrobial activity
Plant species Bacteria Fungi Reference
family part
BUSHES/SHRUBS
Lavandula angustifolia Lamiaceae fs Shigella flexneri Candida albicans Jianu et al., 2013
Lavandula x intermedia Staphylococcus aureus Robu et al., 2016
E. coli
Viburnum opulus Caprifoliaceae hb; fl; fs Staphylococcus aureus Bubulica et al., 2012
Lonicera tatarica Caprifoliaceae hb; fl; fs Staphylococcus aureus Bubulica et al., 2012
Aronia melanocarpa Rosaceae fr; fl Vibrio vulnificus, Giupana et al., 2016
V. cholera, V. mimicus
E. coli
Lycium barbarum Solanaceae fl; fs Staphylococcus aureus Mocanu et al., 2014
Listeris monocytogenes Mocanu et al., 2015c
Bacillus subtilis
Lycium chinense Solanaceae fl Staphylococcus aureus Mocanu et al., 2014
Bacillus subtilis
Listeria monocytogenes
Salmonella thyphimurium
Sambucus ebulus Adoxaceae fr Pseudomonas fluorescens Rodino et al., 2015b
TREE
Juniperus communis Cupressaceae fr Bacillus subtilis Ivopol et al., 2016
Streptococcus luteus
Escherichia coli
Abies alba Pinaceae nd; cx Bacillus subtilis Ivopol et al., 2016
Escherichia coli Sandru et al., 2015
Picea abies Pinaceae nd Staphylococcus aureus Candida albicans Radulescu et al., 2011
Bacillus cereus Aspergillus niger
Proteus vulgaris
Pinus sylvestris Pinaceae nd; cx Bacillus subtilis Ivopol et al., 2016
Escherichia coli
Pinus cembra L. Pinaceae nd; cx Staphylococcus aureus Candida albicans Apetrei et al., 2011
Sarcina lutea Apetrei et al., 2013
Bacillus cereus
Escherichia coli
Fagus sylvatica Fagaceae fl Staphylococcus aureus Pirvu et al., 2014
Robinia pseudoacacia Fabaceae fs; sm Staphyloccous sp., Candida albicans Rosu et al., 2012
cx; fl Streptococcus sp.
E coli., Pseudomonas aeruginosa,
Proteus sp., Salmonella enterica
Cydonia oblonga Rosaceae fl Staphylococcus aureus Cerempei et al., 2016
Escherichia coli
officinalis, in ethanolic and methanolic extracts coli, Pseudomonas aeruginosa, Salmonella

have shown strong inhibitory activity on abony, Staphylococcus aureus, Enterococcus
bacteria and fungi, respectively Escherichia faecalis, Brevibacterium flavum, Sarcina sp.,
675
Bacillus cereus and Aspergillus niger (Soare et was shown by L. chinense extract against
al., 2012). Bacillus subtilis. Meanwhile, extract made of L.
Bushes/Shrubs barbarum flowers was found to be more active
Lavender (L. angustifolia Miller) and lavandin on the Gram-positive bacterial strains; the best
(Lavandula x intermedia) are well known for antibacterial activity was shown against
their medical and cosmetics applications. Staphylococcus aureus (Mocan et al., 2015 c).
Essential oils obtained by steam distillation The perennial herbaceous extracts of Epilobium
from fresh inflorescences harvested in Western hirsutum harvested in Prahova county inhibited
Romania showed antimicrobial activity against both Stapyloccous aureus and E. coli (Pirvu et
Shigella flexneri, Staphylococcus aureus, E. al., 2014). Same group (Pîrvu et al., 2015)
coli and Salmonella typhimurium, while suggested an augmented antimicrobial potency
Streptococcus pyogenes was not sensitive to on Stapyloccous aureus of the combination
their action (Jianu et al., 2013). The authors kaempferol-caffeic acid derivates (aqueous
emphasize the fact that even in the absence of fraction) than myricetin-gallic acid derivate
active principles like linalool and linalyl (ethyl acetic fraction).
acetate, considered responsible for the The dwarf elderberry (Sambucus ebulus) used
antibacterial and antifungal properties of in traditional medicine, has proven to have
essential oils obtained from different species of antibacterial effects on Pseudomonas
Lavandula. This results looks to be in contrast fluorescens and Enterococcus faecalis when
with results reported by Robu et al. (2016) on used as ethanolic extract made of fruits (Rodino
essential oils of lavandin (Lavandula hybrida et al., 2015 b).
Reverchon) harvested in North-Eastern
Romania (Neamt county); these oils showed no Trees
activity against Gram-negative strains; also the Conifers are widely used for the extraction of
results showed that the antistaphylococcal essential oils and their volatile oils contain
activity is reduced, while there is a moderate mainly monoterpene (Ivopol et al., 2016).
antifungal activity. Among the conifers Pinus sp. (Pinaceae) has
Two bushes belonging to Caprifoliaceae been widely studied. Pinus cembra L. from
family, Viburnum opulus and Lonicera tatarica Carpahtian Mountains, bark and needles, have
from Craiova, Dolj county have been tested by antimicrobial effects against Staphylococcus
Bubulica et al. (2012); aqueous extracts of aureus, Sarcina lutea, Bacillus cereus,
aerial parts (stem, flower buds, fruit pulp) have Escherichia coli, Pseudomonas aeruginosa and
been tested on Staphylococcus aureus and Candida albicans (Apetrei et al., 2011; Apetrei
Staphylococcus epidermidis. The results et al., 2013). Common Pinus sylvestris
showed a higher inhibition in the case of essential oils from needles and sprouts showed
Lonicera tatarica extracts. inhibitory effects on Bacillus subtilis,
Studies on bacterial strains isolated from wild Streptococcus luteus, Staphylococcus aureus
birds captured in Danube Delta Biosphere and Escherichia coli; similar results have been
Reservation proved that extracts of fresh fruits obtained by the use of essential oils from
of Aronia melanocarpa has important Juniperus communis berries and Abies alba
inhibition on Vibrio spp. (V. vulnificus, V. needles and sprouts (Ivopol et al., 2016).
cholera, V. mimicus), E. coli, Enterococcus Sandru et al. (2015) also proved inhibition on
faecalis (Giupana et al., 2016). E. coli by the use of Abies alba essential oils.
Ethanolic extracts of Lycium sp. (Solanaceae) Antimicrobial properties of volatile oil is olated
leaves originated in Cluj county, have been from sprouts of Picea abies growing wild in
tested for antimicrobial activity (Mocan et al., Romanian Carpathian Mountains (Prahova
2014). The authors reported that L. chinense Valley) have been tested by Radulescu et al.
extract was more active than L. barbarum (2011). The most evident inhibitory effect was
against both Gram-positive and Gram-negative noticed against the Gram-positive
bacterial strains and that these species as (Staphylococcus aureus, Bacillus cereus),
important sources of flavonoids and Gram-negative (Proteus vulgaris) and fungal
chlorogenic acid. The best antibacterial activity strains (Candida albicans, Aspergillus niger).
676
There are authors which have tested more herbaceous perennial and the rest (25%) are
species from a specific Romanian region for woody perennial species (bushes and trees).
their antimicrobial activity. For example, Pirvu The antimicrobial studies have been mainly
et al. (2014) have focused on herbaceous and focused on species belonging to two botanical
woody plants from Prahova county in families, Asteracea and Lamiaceae, which
propylene glycol solutions or in separate represents 50% of the total studies species.
aqueous, ethyl acetate and chloroform In terms of microbial species can be noticed an
fractions. Among the trees, extracts from leaves intensive focus on pathogenic Gram-positive
of Fagus sylvatica exhibited moderate and Gram-negative bacteria, responsible for
inhibitory effect on Stapyloccous aureus, clinical infections or food contamination. For
ethanolic extracts from flowers and seeds of 89% of the species have been reported
Robinia pseudoacacia have inhibitory activity antibacterial activity, while only 57.8% of the
mainly on Gram-positive coci (Staphyloccous species have proven antifungal activity. The
sp., Streptococcus sp.), while same extracts most reported susceptible fungus was Candida
from bark and leafs inhibited E coli, albicans; few reports are focused on
Pseudomonas aeruginosa, Proteus sp., filamentous fungi like Aspergillus sp.,
Salmonella enterica and Candida albicans. Penicillium sp, Botrytis cinerea or Pythium sp.
As a novelty can be mentioned the use of fall It has been noticed that some of the reports are
quince (Cydonia oblonga) leavesoriginated in novelty in the subject and the researchers have
North-Eastern Romania for the production of approached some spontaneous species little or
natural dye; it has been proven (Cerempei et not ever reported in the international databases
al., 2016) that such dye with mordant (silver (e.g. Helianthemum nummularium, Cydonia
nitrate) have a good antibacterial activity oblonga, Paeonia officinalis). This trend may
against Gram-positive (S. aureus) and Gram- be a solution for further research in the topic, as
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680
TRICHODERMA SPP. SOURCE OF HYDROLASE ENZYMES

WITH ROLE IN F. OXYSPORUM INHIBITION
Mohammed Naithel RADHI1, Cristina PETRIŞOR2, Florica CONSTANTINESCU2,

Ioan ROŞCA1, Hattf Bazool FARHOOD3, Hiba ABD-AL ADHEEM4, Gheorghe STOIAN3
1
2
Research and Development Institute for Plant Protection, 8 Ion Ionescu de la Brad Blvd,
3
University of Bucharest, Faculty of Biology, Department of Biochemistry and Molecular Biology,
91-95 Splaiul Independenței, District 5, Bucharest, Romania
4
University of Thi-Qar, Faculty of Medicine, Nasiriyah, 00964, Dhi Qar, Irak

Abstract
The fungi from Trichoderma genus are a very large group of microorganisms that are present in nearly all agricultural
soils and play a significant role in plant protection. Enzymes produced by species of genus Trichoderma constitute an
important group of biotechnologically enzymes because of the versatility of its properties and ease of mass production.
This study was focused on the effect different factors of antagonistic T. harzianum (ICCF 417) and T. koningii (ICCF
418) against pathogenic fungus F. oxysporum (ZUM 2407) by microbiologic and biochemical tests. There was different
patterns in cell wall degrading enzymes production by Trichoderma isolates so production of chitinase was 113.93 %
by T. harzianum comparative to T. koningii, the lipase was 38.53% in T.harzianum comparative to T. koningii while
the protease was 91.43% in T. harzianum comparative to T. koningii. Both Trichoderma strains studied showed
maximum lipase activity at pH 9, which was significantly higher than those at other pH levels. On the other hand,
optimum pH for producing highest activity of protease was pH 6. A high level of chitinase activity was observed in the
culture medium with pH 6. Our results showed that hydrolase activities studied in this experiment play an important
role in pathogenic fungus F. oxysporum inhibition and the degree of effect is different.
Key words: Trichoderma harzianum, Trichoderma koningii, lipase, protease.
INTRODUCTION implicated in antifungal effect on different

pathogens (Gajera et al, 2012; Vinale et al.,
The use of Trichoderma fungus species against 2007; Harman, 2006; Markovich and
many fungal phytopathogens has seen Kononova, 2003; Viterbo et al., 2002; Monte,
significant progress in recent years because 2001). Several reports have been given to
their high reproductive capacity, ability to explain the role of lytic enzymes produced by
survive under very unfavorable conditions, Trichoderma spp. during plant defense.
efficiency in the utilization of nutrients, strong Fusarium oxysporum, a well-known species of
aggressiveness and efficiency in defense soilborne fungus, causes significant losses to
mechanism (Harman, 2006; Harman et al., horticultural and ornamental crops throughout
2004). Due to their ability to protect plants and the temperate climatic regions (Chen et al.,
because they are commonly found in almost all 2017). The fungus Fusarium spp. is a fungal
soil types, these fungi have been widely studied species that have adapted to different range of
and commercially marketed as biopesticides, environmental from worldwide, in addition this
biofertilizers and soil amendments. types was described as the important pathogen
Trichoderma spp. were shown to be very of the different crops. Lobna et al. (2017) and
efficient producers of extracellular enzymes Houssein et al (2010) found that the use of the
like chitinases, β-1.3-glucanases, cellulases, T. harzianum as strategy to improve the
lipases, proteases some of these have been response of the tomato resistance against
681
Fusarium wilt under greenhouse conditions min., filtered by micro filter, aliquots of the
caused biochemical changes in plants. The supernatant were used for next assays.
correlation between the enzyme level and Characterization of enzymes produced by T.
resistance to the pathogens was increased, koningii and T. harzianum that had effect on F.
especially under conditions of stress and attack oxysporum was done in PDA medium using a
pathogens. Enzymes activity plays an important Petri dishes divided into 6 sections. At the first
role in plant resistance against attacking one we put 10 µL synthetic medium SM, the
pathogens (Zhang et al., 2008; Cherif et al., second 10 µL of dead cell of enzymes culture,
2007; Mohammadi and Karr, 2002). The the third one 10 µL of living cell of enzymes
inhibitory effect of Trichoderma on the culture, the forth section 10 µL of dead cell
phytopathogenic F. oxysporum could be due to from fungi culture, the fifth with 10 µL of dead
the secretion of lipolytic and proteolytic lysate, the last one with 10 µL living lysate.
enzymes that inactivate enzyme of F. For control, we put a Petri dish of T. koningii,
oxysporum thereby decreasing its pathogenity T. harzianum and F. oxysporum alone and a
(Elad and Kapat, 1999). Petri dish divided into two parts, one with
The aim of this study was the evaluation of Trichoderma and the other with F. oxysporum.
different factors from live and dead cells of T. All experiments were made in triplicate.
harzianum, (ICCF 417) and T. koningii strain Chitinase activity assay was performed
(ICCF 418) on hidrolytic enzyme production according to Miller (1959). Lipase activity was
that influence virulence against pathogenic measured spectrophotometrically using an
fungus F. oxysporum strain (ZUM 2407). assay based on the hydrolysis of p-nitrophenyl
palmitate as substrate, according to Gupta et al.
MATERIALS AND METHODS (2002) while for determination of protease
activity has been used the method of Saad
The ability of fungi T. koningii and T. (1995). The protein quantity of the crude
harzianum to produce enzymes were evaluate enzyme extract was determined by the Lowry
in synthetic medium. The fungus was grown in method using bovine serum albumin as
250 ml Erlenmeyer flask that contained 100 ml standard (Lowry et al., 1951). The enzymes
of synthetic medium (SM) then we add the activity and protein concentration were
specific substrates for each enzyme separately, measured after 1, 2, 3 and 4 weeks incubation
then cultures were incubated at 28°C and the of the culture incubation. In order to determine
ability of enzymes production was determined the optimum pH value for the enzyme
after 1, 2, 3 and 4 weeks of incubation. Also produced by Trichoderma obtained after
enzymes were used diluted to 100%, 50% and fermentation, the activity of the enzyme was
10%. The fungi weight was determined by assayed between the pH values of 3.0-9.0.
weighting an empty tube, then put (1 mL) of
the fungi from each period of culture into the RESULTS AND DISCUSSIONS
weighted tube after that drying and centrifuging
and taking the precipitate which was dried at Figure 1 shows the influence of pH on
500C for 3 days, then weighted again. chitinase, lipase and protease production by T.
Preparation of cell lysate was done by taking harzianum, T. koningii and F. oxysporum.
10 ml of each culture separately and According to the results, the optimal pH for
centrifuged. The precipitate obtained was re- lipase was 9. The results are similar with Ulker
suspended in 5 mL of phosphate buffer (pH et al., 2014, which showed the optimum pH
7.4) with vortex, then adding sand to the tube, value for lipase activity produced by T.
put it in IKA® ULTRA TURRAX device at harzianum was 8.5. Also has been establish
6000 rpm for 30 s then put it at ice for 1 min., that optimum pH for maximum chitinase and
replay for three times and centrifuged at 10000 protease activity in T. harzianum and T.
rpm for 10 min. at 4°C. Then the solution koningii was pH = 6. This agrees with results
obtained filtered by Millipore 0.22 µm, and obtained by Cirano et al., 1991, and Kredics et
aliquots of the supernatant were used for next al., 2004.
assays. All cultures were put at 1000C for 15
682
Figure 1. Determination of optimum pH for chitinase, lipase and protease enzymes production
by T. harzianum, T. koningii and F. oxysporum
The results of chitinase enzyme showed that dilution ratio 100% was the highest, as shown
the two fungi can produce the chitinase but its in figure 2.
level on T. harzianum was higher than T. The levels of chitinase enzyme was 0.42 and
koningii, and for the two fungi the period of 0.48 µmol/min in T. koningii and T.
14 days was the highest production, also the harzianum, respectively, after 14 days.
Figure 2. Chitinase production in T. koningii and T. harzianum in a different dilution ratio of enzyme
and for a period 1, 2, 3 and 4 weeks
The production of lipase enzyme in synthetic highest period of production, as shown in

medium for T. koningii was higher than T. figure 3.
harzianum and the second week was the
683
Figure 3. Lipase production in T. koningii and T. harzianum in a different dilution ratio of enzyme
The results of protease production by T. incubation period was with the highest
koningii and T. harzianum are presented in production, also the dilution ratio 100% was
figure 4. The protease levels in T. koningii was the highest increasing.
higher than in T. harzianum, and the 30 days
Figure 4. Protease production in T. koningii and T. harzianum in a different dilution ratio

of enzyme and for a period 1, 2, 3 and 4 weeks
The protein levels in chitinase enzyme were in the second week had the highest levels of
T. harzianum higher than the T. koningii, and protein in the two fungi, as shown in figure 5.
Figure 5. Protein levels in T. koningii and T. harzianum in a different dilution ratio of enzyme
The highest protein level in the lipase enzyme 14 days was the highest level of protein
medium was in T. koningii in comparison with production in comparison with the rest periods,
its level in T. harzianum, but also the period of as shown in figure 6.
684
Figure 6. Protein levels in T. koningii and T. harzianum in a different dilution ratio of enzyme
In figure 7 are presented the levels of proteins protein levels on T. koningii was higher than T.
in synthetic medium for protease production by harzianum, and for the two fungi the period of
in T. koningii and T. harzianum respectively, 30 days was the highest production.
Figure. 7. Protein levels in T. koningii and T. harzianum in a different dilution ratio of enzyme
The levels of protein in T. koningii were 52.97, lipases secreted by Trichoderma spp. may be
101.85 and 155.23 mg/mL for chitinase, lipase involved in inactivating extracellular enzymes
and protease, respectively, in out of cell, while of F. oxysporum.
in cell lysate were in chitinase 36.98 mg/mL,
lipase 65.69 mg/mL and protease 75.68 CONCLUSIONS
mg/mL. In T. harzianum the protein levels The results obtained in this study show that
were 67.85, 72.02 and 99.25 mg/mL for both strains of Trichoderma studied produced
chitinase, lipase and protease, respectively, in chitinases, proteases and lipases in synthetic
out of cell, while in cell lysate were in chitinase medium but the concentration depends on the
45.26 mg/mL, lipase 31.68 mg/mL and strain and the incubation period.
protease 58.61 mg/mL. The results of the effect There is an inhibitory effect of T. harzianum,
of T. harzianum and T. koningii on F. strain ICCF 417 and T. koningii strain ICCF
oxysporum showed that these fungi have a high 418 against pathogen F. oxysporum strain
ability against the pathogenic fungi F. ZUM 2407 by production of protein and
oxysporum, the ratio reached to 1, according to extracellular enzymes which may effect on the
the scale of Bell et al. (1982). The lipase and action F. oxysporum.
the protease enzymes were effective on F. The lipase has the highest weight in
oxysporum in the second and forth weeks, comparison with the others enzymes, while the
respectively. These results are in accordance chitinase enzyme was not effective on F.
with those obtained by Elad et Kapat, 1999, oxysporum for all period of incubation.
which supported that some proteases and
685
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P.Y., Guan Z.H., Qin H.Y., Liu A.R., Xiao- Min Lin tomato cultivars with differing level of resistance to
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Chérif M., Arfaoui A., Rhaiem A., 2007. Phenolic of Trichoderma and their role in plant defense from
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of chickpea to fungal pathogenic attacks. Tunisian Microbiology, 39 (4): 341-351.
Journal of Plant Protection, 2: 7-21. Miller G.L., 1959. Use of dinitrosalicylic acid reagent for
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chitinase synthesis in Trichoderma harzianum. Chemistry, 31 (3): 426-428.
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Elad Y., Kapat A., 1999. The role of Trichoderma chitinase activities in soybean root nodules. J. Plant
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Gajera H.P, Bambharolia R.P., Patel S.V., Khatrani T.J., Trichoderma: Between Agricultural Biotechnology
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686
EFFECT OF PERIODS AND REPETITION OF THE BURNING

METHOD IN THE CONTROL OF WATER HYACINTH
(EICHHORNIA CRASSPIES) PLANT
Al-Gburi Omar ABDULRAZZAQ SHIHAP1, Talal Abed KHATTAB1, Cristina PETRIŞOR2

1
University of Mosul, Faculty of Agriculture and Forestry, Mosul No.80, Irak
2
Research and Development Institute for Plant Protection, 8 Ion Ionescu de la Brad Blvd,

Abstract
The experiment carried out using basins under field conditions of the Faculty of Agriculture and Forestry, University of
Mosul in the season 2010-2011. The aim of study is using the mechanical method (burning method) for controlling
water hyacinth and decrease the pollution on aquatic environment. The study included a two factors, influence burning
repetition and periods of the burning in the control of water hyacinth weed. The experiment was applied in randomized
completed design it was applied in three repeaters. The results showed the no influence on studied traits in the case of
an incineration process one time while the increase in the number of times the incineration twice to a negative impact
on the studied traits of the water hyacinth plant this effect was more severe in the event of an increase in the replicates
of burning to three times, with low in the all studied traits (length of plant, the number of leaves, length of spike,
number of flowers and weight wet and dry weight the total vegetation and roots).
Key words: water hyacinth, burning, repetition burning.
INTRODUCTION impeded the work of the hydro power and

pumping stations as well as a haven for many
Water hyacinth of floating plants on the surface of the diseases and insects in the depletion of
of the water and usually floats by rafts publish oxygen dissolved in the water, which
on the water surface, which consists of a roots negatively affects life's bountiful fish and some
group under the water's surface and total of the people who depend on fishing. In order
vegetative floats above the surface of the water, to reduce the pollution caused by the excessive
there are several names for this plant, including use of herbicides and non-compliance with the
the flower Nile, yanst water or herb Nile control programs and the recommended
(Eichhornia crassipes (Mart.) Solms), belongs concentrations used for control, the burning is
to the family Pontederiaceae, also called family the one of mechanical methods used to combat
Pickerel weed family. Belong to this family six weed widespread in agricultural and non-
genera and about 20 species, and the plants of agricultural land. There are several methods
this family are either floating on the surface of that have been used to combat this weeds in
the water or floating and flourish in the warmer many countries where preventive, mechanical,
regions of the globe. There are several species biological and chemical methods have been
of the genus Eichhornia most important (E. deployed. So researchers and studies in this
crassipes ) for being a dangerous weed, which field are interested in working hard in the
occupies the eighth spot (Holm et al., 1977). preparation of control programs in order to
The ornamental plant enter in Irak in mid 1980s limit the spread and control of the weed, its
because it have purple flowers and some civil money has an impact on the human and its
nurseries selling this plants as ornamental plant sources of livelihood and other resources. Due
and they spread to the rest of the other areas to the lack of sources on the use of the method
currently present in separate regions in Mosul, of burning in the control against the water
and speed spread and growth of it led to more hyacinth, it was pointed out that this method
problems impeding the flow of water in was used with the reed plant, Common reed,
irrigation channels and drainage and also Phragmites australis (Cav.)Trin., because it is
reduced quantities of water consumption and in an environment similar to the environment of
687
the water hyacinth plant. Burning in general field of the Faculty of Agriculture and Forestry,
does not completely reduce the ability of wild University of Mosul on 3.03.2010, three plants
reed to grow because it is difficult to burn roots were taken from the basin, which was prepared
and rhizomes which are often covered with a in the field length of 15 m and width 6 m in
thick layer of soil or soil and water (Beall, each pot, which number 30 and dimensions 73
1984). Of course, this result applies to the water cm length and width 32 cm and depth of 33 cm.
hyacinth, where if burned green parts and This element was covered with thermal
leaves, it may be difficult to reach the burning insulated edges of aluminum to preserve the
to some of the leaves and roots because they element of heat consisting of the burning
are covered with a layer of water, which leads process and the burn tool is a regulator + gas
to the re-growth of these parts. Several studies tube + burner.
have indicated that the date of burning of cane Where the factors of the experiment included
has an effect on plant density and growth. one factor is the factor of burning times and
Burning at the end of summer is better than repetition as follows:
burning in winter and spring (Cross and 1. Burning one after month of comparison
Fleming, 1989). In an experiment conducted by in day 12.07.010.
(Thompson and Shay, 1989) in the Marsh 2. Burn one after a month + burn again
Marshes (Delta March Manitoba), he found that after 10 days of the first burn.
burning in spring, summer and fall leads to a 3. Burn one after a month of comparison +
higher growth rate in the vegetative growth but burn three after 20 days of the first
shorter and thicker but the biomass of the burn.
vegetative group was greater for the plants 4. Burning one after two months of
burned In the spring and fall while they are comparison on day 12.08.2010.
lower in plants that were burned in the summer. 5. Burn one after two months of
This is due to the fact that plants whose growth comparison + burn the second after 10
season was in the spring and fall gave them the days of the first burn.
opportunity to grow well. Burning leads to a 6. One burn after two months of
doubling of the size of the biomass on the comparison + burning three after 20
surface of the earth due to the availability of days of the first burn.
light and soil ventilation around the rhizomes 7. Burning one after three months of
(Graneli, 1989). The same researcher comparison on day 12.09.2010.
recommended that the burning of cane plants in 8. Burn one after three months of
winter causes slight damage to plants, while the comparison + burn second after the 10
date of burning in the period of emergence of days of the first burn.
plants leads to the killing of the total vegetative 9. Burn one after three months of
almost completely. (Uchytil, 1992) found that comparison + burn the second after 20
burning of wild reed plants when growing well days of the first burn.
and intensely is more efficient, the high heat 10. Comparison (without burning).
resulting from burning vegetation also causes Basins were distributed at random within the
damage or death to reed rhizomes found design (CRD) and with three replicates. The
beneath the surface of the soil. The objective of water was added twice a week to complete the
the research is to study the effect of the water volume in the basin to 77088 cm3, using
recurrence of the date of burning in the fight the same water source used in the first
against the water hyacinth, as well as reduce experiment. A basin was randomly selected for
the percentage of pollution caused by the use of each treatment and burned by the burner for
chemical herbicides in control. one minute and repeated three times. The
experiment was analyzed according to the
MATERIALS AND METHODS random complete design of simple experiments
(CRD). I used the computer according to the
Effect of repeated burning at different dates (SAS) program and used the Dunkin Multi-
in the controlling of water hyacinth Range Test to compare (Al-Rawi and Abdul-
This experiment was carried out in basins Aziz, 1980).
where it was placed under the conditions of the
688
RESULTS AND DISCUSSION The length of the inflorescence (cm) and the
number of flowers/plants of the water hyacinth
Plant height (cm) plants were not significantly affected compared
with the comparison treatment (Table 1). In the
Table 1 indicates that the height of the Nile
case of plant age progress after the burning
flower plants was not significantly affected by
process after a month or two months and
the one-time burning process and its various
repeated burning twice resulted in a significant
dates (12.07., 12.08. and 12.09.2010). In the
decrease for two adjective (61.80, 63.87, 54.72
case of repeated burning two times, the same
and 54%) for both the two adjective after a
table indicated that the process of burning at
month or two months of burning twice
the first date of the plant life (22.07.2010) and
compared to Comparative Plants. In the case of
twice did not significantly affect the attribute of
repeated burning three times and their different
plant height, but the progress of the plant age,
dates, it all these treatments significantly
after the process of burning after a month or
reduced the length of the inflorescence (cm)
two months and the repeat of burning (55.72
and the number of flowers/plant of the water
and 52.18%) treatment of burn twice and for
hyacinth (37.57, 75.77, 100, 45.45, 72.73 and
the second and third seasons, respectively,
100%) for the third burn with their different
compared to the plants that were not treated by
dates and the two adjective, respectively,
burning. This is due to the fact that the burning
compared with non-treated plants.
process was carried out at the stage where the
Number of leaves/plant, leaves area (m2/m2)
plant during this period in the stage of active
It is noted from the same table that the process
growth and despite the influence of this process
of burning one time at the date (12.07.2010)
in total vegetative the effect of burning was
did not significantly affect the number of
temporary, as the stolon and daughter plants
leaves/plant as well as the of leaves area
were able to form new growths and daughter
(m2/m2) compared with the treatment of
plants new where they were about the same
comparison, but it was observed there is a
height as those plants that were not treated with
significant effect of the two adjectival on the
burn. The results were taken at the end of the
burning date (12.08. and 12.09.2010). Morally
experiment, after about eight months. This
decline was observed of these two adjectival
period is enough for a plant to give these
(43.5, 38.83, 35.97 and 23.17%) for both the
growths. As for the plants treated late in the
two dates respectively. In the case of repeated
treatment, it seems that the plants treated with
burning of two times, the same table indicated
burning were in a good growth stage and so the
that the process of burning on the first date
burning process led to significant damage in the
(22.07.2010) and twice did not significantly
vegetative group and when the plant returned to
affect the number of leaves/plant of the water
grow again there was a second burning, which
hyacinth compared with the comparison
led to the weakening of the plant, negatively
treatment, but it was observed there is a
affect the height of these plants during the time
significant effect of the leave's area in this date,
period between the treatment procedure and the
where the decrease was 29.27% compared to
period of termination of the experiment
the unburned plants. In the increase plant age,
compared with the comparison treatment. In the
after burning after a month or two months and
case of repeated burning of three times and
repeated burning twice, resulted in a significant
their different dates, all these factors have
decrease in the number of leaves/plant and the
resulted in a significant decrease in plant height
leaves area (m2/m2) of the water hyacinth by
(61.34, 77.87 and 76.64%) for the burning
64.67, 79.5, 75 and 84.15% after month or two
processes (2.08., 2.09. and 2.10.2010) which
months after burning twice compared to the
were burned three times respectively.
Plants that did not burn. This may be due to the
Length of inflorescence (cm) and number of
fact that the plants that were burned by a late
flowers / plant
date were in the growth and total good
The process of burning one time and its
vegetative and when treated with burning, this
different dates (12.07. and 12.08. and
process led to significant damage to the
12.09.2010) and also when the burning
vegetative group and when the plant to
(22.07.2010 + 10 days of the first heartburn)
689
compensate the growth again, there was a plant. the case of repeated burning of the three
second burning, which led to the depletion of times and their different dates, all these
food stored in places Plant storage, which led to treatments resulted in a decrease in the number
the weakening of the plant, which adversely of leaves/plant as well as the leaves area of the
affected the number of leaves/plant, which in plant (m2/m2) by (37.17, 71.17, 97 and 51.22%)
turn affected the leaves area (m2/m2) of this to burn three times.
Table 1. Effect of the date and repeated of burning in the traits studied Plant water hyacinth growing
in the site of the Faculty of Agriculture and Forestry University of Mosul for the season
Thoughtful adjective
Plant Number Leaves Length of Flowers Fresh weight Dry weight of Fresh eight Dry weight
height / of area Inflorescence number/ of vegetative the vegetative of root of the root
cm leaves (m2/m2) (cm) plant total total (kg/m2) (kg/m2)
(kg/m2) (kg/m2)
Comparison (without 38.10a 600 a 1.64 a 15.97 a 11 a 22.59 a 2.40 a 8.69 a 1.23 a
burning).
Burning one after month of 37.63 a 615 a 1.84 a 14.80 ab 9 ab 19.63 ab 2.41 a 7.62 ab 1.40 a
comparison in day
12.07.2010.
Burn one after a month + 30.00 a 454 ab 1.16 b 13.17 ab 8 ab 13.79 c 1.74 b 7.95 ab 0.98 a
burn again after 10 days of
the first burn.
Burn one after a month of 14.73 b 377 bc 0.80 e 9.97 bc 6 cd 9.27 d 0.83 cd 5.94 b 0.77 a
comparison + burn three after
20 days of the first burn.
Burning one after two months 32.63 a 339 bc 1.05 de 13.50 ab 9 ab 13.73 c 1.33 bc 6.80 ab 19.44 a
of comparison on day
12.08.2010.
Burn one after two months of 16.87 b 212 cd 0.41 f 6.10 cd 3 cd 4.61 ef 0.27 de 2.53 c 0.23 a
comparison + burn the second
after 10 days of the first burn.
One burn after two months of 8.43 b 173 cd 0.47 f 3.87 de 3 cd 2.52 f 0.15 e 1.35 c 0.12 a
comparison + burning three
after 20 days of the first burn.
Burning one after three 28.87 a 367 bc 1.26 b 13.97 ab 11 a 18.05 b 1.86 ab 7.73 ab 0.90 a
months of comparison on day
12.09.2010.
Burn one after three months 8.90 b 123 d 0.26 f 5.77 cd 5 bc 6.73 de 0.54 de 2.21 c 0.27 a
of comparison + burn second
after the 10 days of the first
burn.
Burn one after three months 18.22 b 18 d 0.00 g 0.00 e 0.00 d 0.45 d 0.03 e 0.32 c 0.04 a
of comparison + burn the
second after 20 days of the
first burn.
The values followed by similar letters are not significantly different from each other at the 5% probability level in each of the study factors and their
interference.
Fresh and dry weight of vegetative total equal or equal to those plants not treated by
(kg/m2) burning. In the case of progress of the plant
The adjective of fresh and dry weight of age and after one or two months of this burn,
vegetative total (kg/m2) Plants of the water there is a significant effect for the two
hyacinth table (1) were not significantly adjective and decrease by (39.22, 20.10 and
affected when you make the burning process 44.58%) for both the two adjective and both
once the deadline (12.07.2010) as well as at others time respectively. It may be because
the date (12.09.2010) for the weight dry for the burning process was carried out in the
vegetative total, the reason is that a burning case of the plant is very activity where it had
process in the early stages of the experiment a large number of vegetative leaves so many,
may cause damage to the vegetative total, but the process of burning in this period leads to
these plants soon quickly re-growth as a result significant damage to the total vegetation so
of the growing vegetative by daughter plant or that some of these plants, which was able to
stolon in this period the plant active in re-growth Were not able to form a total
produce many new growth Therefore it was vegetative equal in these treatments were not
able to reach the fresh weights of almost treated by burn during the period of time the
690
experiment was harvested, which was less incineration factors and their different dates
than the period during which the burning compared with the plants that were not treated
process was carried out at the beginning of with burning.
the experiment. In the case of repeated
burning of two times, three times and their CONCLUSIONS
different dates, all the treatments resulted in a
significant decrease in the fresh and dry Delaying the time of burning is favoring for
weight of the total vegetation of the water the control of water hyacinth plant.
hyacinth plant (38.95, 79.59, 70.21, 27.5, More cover the results of present study
88.75, 77.5, 58.96, 88.84, 98.01 and 65.42, confirmed that increasing the frequency of
93.75 and 98.75%) the burn two time and burning more than twice gave an excellent
three time, for both the different s dates results for the controlling of this weed which
respectively, compared to non-burning plants. reflexed in the reduction of all characteristics
Table 1 shows that the one-time burning studied.
process (12.07., 12.08. and 12.09.2010) did
not significantly affect the adjective fresh REFERENCES
weight of root total of the water hyacinth
plant. In the case of repeated burning two Al-Rawi K.M., Abdul-Aziz M.K.A., 1980. Design and
analysis of agricultural experiments, National
times, the same table indicated that the Library for Printing and Publishing the Directorate
process of burning at the date (22.07.2010) of University of Mosul, the Ministry of Higher
and twice did not significantly affect the fresh Education and Scientific Research, Irak.
weight of the total root of the plants of the Beall D.L., 1984. Brigantine Division-Marsh
water hyacinth plant, but in the case increase vegetation rehabilitation - Chemical control of
Phragmites. U.S. Fish and Wildlife Service. 8PP.
plant age after burning after a month or two Cross D.H., Fleming K.L., 1989. Control of
months and In the case of repeated burning, Phragmites communis or common reed. U.S. Fish
this process resulted in a significant decrease and Wildlife Leaflet 13.4.12.5.
in the fresh weight of root of the plant by Graneli W., 1989. Influence of standing litter on shoot
(70.89 and 74.57%) for both the two times production in reed, Phragmites australis (Cav.)Trin.
Ex Steudel. Aquatic Botany, 35: 99-109.
respectively, compared to the plants that did Holm L.G., Plucknett D.L., Pancho J.V., Herurger J.P.,
not burn. While when the repeated burning 1977. The World’s Worst Weeds, Distribution and
three times, for different times in all the Biology. The University Press of Hawaii, 609.
treatments, there was a significant decrease in Thompson D.J., Shay J.M., 1989. First-year response
the fresh weight of the root group of the Nile of a Phragmites communis marsh community to
seasonal burning. Can. Journal Botany, 67: 1444-
flower plants by (31.64, 84.46 and 96.32%) 1455.
for the burning three times and different Uchytil Rj., 1992. Phragmites australis. In: Fire
dates, respectively compared with the natural Effects Information System, [Online]. U.S.
plants. It was also observed that there was no Department of Agriculture, Forest Service, Rocky
significant difference between recurrence of Mountain Research Station, Fire sciences laboratory
(producer). Available: 19
burning two and three times when compared http://www.Fs.Fed.us/database/Feis/[2007,
between them. The dry weight of the root february19].
plant of the water hyacinth was not
significantly affected by the various
691
692
BAKERY PRODUCTS FORTIFIED WITH DRIED FRUITS

OF ARONIA MELANOCARPA
Monica CATANĂ1, Luminița CATANĂ1, Enuța IORGA1, Adrian Constantin ASĂNICĂ2,

Nastasia BELC1
1
National Research and Development Institute for Food Bioresources, IBA Bucharest,
6 Dinu Vintilă, District 2, 021102, Bucharest, Romania
2
Faculty of Horticulture, 59 Mărăşti Blvd, District 1, 011464, Bucharest, Romania
Abstract
Aronia (Aronia melanocarpa) has gained a huge interest due to its complex biochemical composition that gives it
various beneficial effects on health. Polyphenols (anthocyanins and procyanidins, especially), represent the most
important group of biologically active compounds, which give to these fruits their therapeutical properties and
antioxidant potential. In this study it was evaluated the quality of bakery (bread, minibaguette and biscuits) products
fortified with dried fruits of Aronia melanocarpa. Products made have superior sensory quality, high nutritional value
and antioxidant potential. Thus, products are characterized by the content in polyphenols (193.34 ... 263.22 mg
GAE/100g), proteins (11.92 ... 12.95%), fibres (9.13 … 16.60%) and mineral elements (potassium, calcium,
magnesium, iron and zinc). Antioxidant capacity of the achieved products varied in the range 1.91 ... 3.42 mg Trolox
Equivalents/g. Biscuits with Aronia and ginger recorded the highest value of antioxidant capacity, and Bread with
Aronia had the minimum value. Shelf-life of products studied is quite long, possibly due to the antioxidant and
antibacterial properties of fruits of Aronia melanocarpa. In addition, in case of bread and minibaguette, lactic acid
bacteria from sourdough used for fermentation have an important role to ensure the preservation of these products.
This study has practical value; dried fruits of Aronia melanocarpa can be a functional ingredient to increase the
nutritional value and antioxidant capacity of bakery products.
Key words: Aronia melanocarpa, polyphenols, antioxidant capacity, bakery products.
INTRODUCTION substances which give the antioxidant potential

of black chokeberry fruits (Kokotkiewicz et al.,
There are scientific evidences that a diet rich in 2010). Total polyphenolic content varies in the
fruits and vegetables may reduce the risk to range 2-8 mg/100 d.m. and depends on the
have different chronic diseases (Borges et al., cultivar, growing conditions and harvesting
2010). Berries are recommended in a healthy time (Kähkönen et al., 1999; Hudec et al.,
diet as it provides protection against degene- 2006; Oszmiański and Wojdyło, 2005;
rative diseases, cardiovascular diseases and Benvenuti et al., 2004; Sueiro et al., 2006;
cancer (Howard et al., 2012). This protective Hakkinen et al., 1999). Jakobek et al. (2012)
role is given by some biologically active determined total polyphenolic content in case
compounds they contain, like: phenolic acids, of three cultivars (ʻVikingʼ, ʻNeroʼ,
anthocyanins and flavonoids (De Pascual- ʻGaliciankaʼ) of fruits of chokeberry (Aronia
Teresa et al., 2008). Among berries, fruits of melanocarpa) and wild chokeberries, in
Aronia melanocarpa, they have gaine recently Croatia, region Slavonia during two
attention due to the health claims associated consecutive years (2010 and 2011). Cultivars
with their consumption (Chrubasik et al., 2010; ʻVikingʼ, ʻNeroʼ and wild chokeberries had a
Kokotkiewicz et al., 2010). similar total polyphenolic content (9012–
Fruits of Aronia melanocarpa (Michx.) Elliott 10,804 mg kg−1 in the first year, 9361–12,055
are rich sources of biologically active mg GAE/ kg FW in the second year). Cultivar
compounds, polyphenols (anthocyanins and ʻGaliciankaʼ had a lower total polyphenolic
procyanidins, especially) representing the most content (8564 mg GAE/kg−FW first year, 8600
important group. Polyphenols are the main mg GAE/kg FW second year).
693
Besides polyphenols, fruits of Aronia In this paper are presented bakery products
melanocarpa are sources of sugar (10-18%), („Bread with Aronia”, „Minibaguette with
pectins (0.6-0.7%), the sugar alcohol sorbitol, Aronia”, „Biscuits with Aronia and cinnamon”,
and parasorboside (Wolski et al., 2007; „Biscuits with Aronia and ginger”) fortified
Niedworok and Brzozowski, 2001; Weinges et with dried fruits of Aronia melanocarpa. Pro-
al., 1998; Kulling and Rawel, 2008). ducts quality was evaluated through sensory,
Fresh fruits of Aronia melanocarpa can be physico-chemical and microbiological analy-
consumed a short period time and thus to ses. Also the antioxidant potential was evalua-
benefit by their nutritional qualities and ted by determination of total polyphenolic
antioxidant potential these fruits are processed content and of antioxidant capacity.
under various forms: dried fruits, puree, juice,
liqueur, syrup, jam, wine, compote, tea, powder MATERIALS AND METHODS
(Chrubasik et al., 2010; Ochmian et al., 2012;
Kapci et al., 2013; Šnebergrová et al., 2014). Materials
On the other side, fresh fruits of Aronia Dried fruits of Aronia melanocarpa used within
melanocarpa, have sour and astringent taste experiments were obtained from organic
and therefore consumers prefer juice of Aronia culture (Figure 1). These fruits were ground
melanocarpa, in combination with other fruits, with a Retsch mill within the performed
such as, apples, pears and blackcurrant experiments. The raw materials and materials
(Lehmann, 1990; Ara, 2002). used for making bakery products were
Kapci et al. (2013) investigated the antioxidant purchased from the market.
potential of fruits of Aronia melanocarpa and
of their derivate products (Chokeberry juice,
Chokeberry pomace, Chokeberry concentrate,
Chokeberry syrup, Chokeberry compote,
Chokeberry jam, Raspberry-chokeberry syrup,
Sour cherry-chokeberry syrup). Total poly-
phenolic content varied in the range: 0.78 ±
0.02 g GAE/kg ... 63.1 ± 0.5 g GAE/kg
(minimum value was recorded by Raspberry-
chokeberry syrup and the maximum one by Figure 1. Dried fruits of Aronia melanocarpa
Chokeberry pomace, due to the fact that it
contains skin and seeds of chokeberry). It Methods
should be noted that dried chokeberry have a Sensory analysis
high total polyphenolic content (39.9 ± 0.3 g Sensory evaluation of the bakery products
GAE/kg, respectively, 50.1 ± 0.4 g GAE/kg). fortified with dried fruits of Aronia
Antioxidant capacity of chokeberry fruit deter- melanocarpa was performed 12 hours after
mined by ABTS, DPPH, and CUPRAC were baking, using „Comparison method with
10.9 g·kg-1, 11.3 g·kg-1 and 67.7 g·kg-1, res- unitary score scales”. Sensory quality of the
pectively. By all methods, the highest antio- fortified product was established based on the
xidant capacity was recorded in case of dried total average score by comparison with a scale
chokeberries (54.4 ± 1 g·kg-1 by ABTS, 30.5 ± from 0 to 20 points (18.1 ... 20 - qualyfing
1 g·kg-1 by DPPH and 233.2 ± 1.3 g·kg-1 by „very good”; 15.1 ... 18 - qualyfing „good”;
CUPRAC). The lowest antioxidant capacity 11.1 ... 15 - „satisfactory”; 7.1 ... 11 -
was recorded in case of Raspberry-chokeberry „unsatisfactory”; 0 ... 7 - „inadequate”).
syrup (0.7-1.2 g·kg-1), the obtained results For measurement of colour parameters dried
correlated with content in total polyphenols, fruits of Aronia melanocarpa were ground with
total flavonoids, total anthocyanins. Retsch mill, and bakery products fortified were
Taking into account the high content of liophylised and then grounded with a Retsch
biologically active compounds and antioxidant mill. Measurement of the colour parameters of
capacity of dried fruits of Aronia melanocarpa, samples was performed at room temperature,
their use to fortify food products has a real using a HunterLab colorimeter, equipped with
interest. Universal Software V4.01 Miniscan XE Plus
694
programme, to register CIELab parameters (the achieved in the concentration range 0 to 0.20
Commission Internationale de l’Eclairage - mg/mL. The extraction of phenolic compounds
CIE), L*, a* and b*: L* - colour luminance (0= was performed in methanol: water = 50:50 and
black, 100 = white); a* - red-green coordinate the absorbance of the extracts were determined
(-a = green, +a = red); b* - yellow-blue at a wavelenght λ = 755 nm. Results were
coordinate (-b = blue, +b = yellow). expressed as mg of Gallic Acid Equivalents
The texture properties of the bakery products (GAE) per g product.
fortified with dried fruits of Aronia
melanocarpa were measured through a Antioxidant capacity
compression test using an Instron Texture The DPPH scavenging radical assay was
Analyzer (model 5944, Illinois Tool Works conducted according to Horszwald and
Inc., USA). Andlauer (2011) with some modifications
(concerning extract volumes of the used sample
Physic-chemical analysis and reagents, using UV-VIS Jasco V 550
The moisture content was determined accor- spectrophotometer). The reaction was perfor-
ding to the ААCC 44-15A method. Protein med in dark for 30 min (at ambient tempe-
content was determined by the Kjeldahl method rature) and after this time the absorbance was
with a conversion factor of nitrogen to protein read at 517 nm. It was achieved the calibration
of 6.25 (AOAC Method 979.09, 2005). Fat curve Absorbance = f (Trolox concentration),
content was determined according to AOAC in the concentration range 0-0.4375 mmol/L
Method 963.15, and ash content according to and the results were expressed as mg Trolox
AOAC Method 923.03 (AOAC, 2005). Mineral Equivalents per g product.
elements content was determined by atomic
absorption spectrophotometer (type AAnalyst Microbiological analysis
400, Perkin-Elmer, Waltham, MA, USA) from The water activity (Aw) was determined by an
HCl mineralized sample. instrument Aquaspector AQS-2-TC, Nagy. The
Total dietary fiber (TDF) was determined by measurements were performed at 25ºC. Yeasts
enzymatic method using the assay kits: K- and molds were determined by the method SR
TDFR „Total dietary fiber” (AOAC Method ISO 21527-1: 2009. Enterobacteriaceae were
991.43). determined according to the SR ISO 21528-2:
Total sugar and reducing sugar content was 2008 method and Escherichia coli by SR ISO
determined according to Schoorl method. For 16649-2: 2007 method. Salmonella was
total sugar, the method is applied after an acid determined by the method SR EN ISO
hydrolysis (20% HCl solution) at 70ºC, for 27 6579:2003/AC: 2006.
minutes.
Calorie contents were calculated using the RESULTS AND DISCUSSIONS
following conversion factors: 9 for fat, 4 for
carbohydrates, 4 for protein and 2 for fibre, Sensory analysis
according to the Commission Regulation no. Sensory analysis plays an important role in
1169/2011 (European Commission, 2011). characterizing the quality of food products.
Joule contents were calculated using the Results of sensory analysis of bakery products
following conversion factors: 37 for fat, 17 for showed that the addition of powder of Aronia
carbohydrates, 17 for protein and 8 for fibre, melanocarpa, in their composition, has not a
according to the Commission regulation no. negative effect on sensory characteristics
1169/2011 (European Commission, 2011). (Figure 2).
Total polyphenol content So, the analyzed products were tested by an
Total polyphenol content was conducted expert panel receiving qualifying „very good",
according to Horszwald and Andlauer (2011) with scores in the range 19.44-19.92, as
with some modifications (concerning extract follows: Biscuits with Aronia and cinnamon-
volumes of the used sample and reagents, using 19.44; Biscuits with Aronia and ginger-19.52;
UV-VIS Jasco V 550 spectrophotometer), Bread with Aronia and seeds-19.76; Miniba-
based on calibration curve of gallic acid guette with Aronia and seeds-19.92 (Figure 3).
695
Using of sourdough for fermentation and final Aronia” (L* = 40.18). In comparison with the
proofing, in case of bread and minibaguette, achieved products, Aronia fruits have the most
give them an elastic and dense crumb, proper intense colour (L* = 18.55) (Figure 4).
texture and in the same time intense and
pleasant flavour.
Figure 4. Colour parameters of Aronia fruits

and bakery products fortified with Aronia
In the case of the four bakery products, the

colour parameter a* recorded close values
(7.10 ... 7.84), while the colour parameter b*
varied in the range 7.45 ... 10.58 (the minimum
value was recorded for the product
„Minibaguette with Aronia” and the maximum
one for the product „Biscuits with Aronia and
cinnamon”).
Texture properties of bakery products fortified
with dried fruits of Aronia melanocarpa,
during the shelf-life, packed in polypropylene
film are presented in Tables 1 and 2.
Figure 2. Bakery products fortified with Aronia Table 1. Texture properties of products Bread with
Aronia and Minibaguette with Aronia
Table 2. Texture properties of products Biscuits

fortified with Aronia
Figure 3. Sensory evaluation of bakery products
fortified with Aronia
According to the results obtained, the darkest

colour was recorded for the product „Biscuits
with Aronia and ginger” (L* = 29.6), and the
least intense for the product „Bread with
696
Bread with Aronia had the lowest firmness, in Table 3. Chemical composition (%) of bakery products
comparison with those of Minibaguette with fortified with Aronia
Aronia, due to the highest moisture content
(41.69 ± 0.30%). During the 7 days firmness
varied in the range 4.23 ... 10.38 N, in case of
Bread with Aronia, and, respectively, 9.98 ...
30.44 N, in case of Minibaguette with Aronia.
Elasticity and, respectively, cohesiveness, have a
Expressed as kcal/100 g product.
values relatively close in case of those two b
Expressed as kJ/100 g product.
bakery products. Although initially those two
assortments of biscuits with Aronia have close Also biscuits can have a beneficial effect on
values of firmness and brittleness, after 72 days diabetes because they contain ingredients with
of storage, Biscuits with Aronia and ginger hypoglicemiant effect: fruits of Aronia
have firmness of 1.84 times higher in melanocarpa, ginger and cinnamon (Simeonov
comparison with Biscuits with Aronia and et al., 2002; Valcheva-Kuzmanova et al., 2007;
cinnamon, and brittleness of 1.28 times. This Li et al., 2012; Lu et al., 2012).
can be explained by difference in lipid content, Due to the used raw materials, the bakery
water content and, in the same time, by products fortified with Aronia have high
difference in composition between those two mineral element content (Table 4). The most
biscuit assortments. abundant among investigated elements was
potassium, its concentration varying in the
Physic-chemical analysis range 4784,60 … 10547 mg/kg (maximum
Dried fruits of Aronia melanocarpa, used to concentration was recorded for Minibaguette
fortify bakery and pastry products, have a with Aronia).
complex biochemical composition, by total
polyphenolic content, mineral elements and Table 4. Mineral element content of bakery products
fibre content, especially. Their total fortified with Aronia
polyphenolic content was 3180.90 ± 84.67 mg
GAE/100 g, comparable with those reported by
Kapci et al. (2013): 3990 ± 30 mg GAE/100 g,
5010 ± 50mg GAE/100 g, respectively. Also,
fruits used within experiments had antioxidant Also, the achieved products have high calcium
capacity of 67.29 ± 2.82 mg Trolox/g, higher content, which varied between 1100.29 mg/kg
than those reported by Kapci et al. (2013): 36.3 (for Bread with Aronia) and 1691.4 mg/kg (for
± 1.2 mg Trolox/g, respectively, 30.5 ± 1.0 mg Biscuits with Aronia and ginger), being signifi-
Trolox/g. In the same time, dried fruits of cantly higher than those obtained by Ajibola et
Aronia are an important source of potassium al. (2015) in case of biscuits prepared from
(9693.3 ± 1095.34 mg/kg), calcium (1563.7 ± different blends of whole-wheat flour, Moringa
161.22 mg/kg), magnesium (694.1 ± 109.95 oleifera leaves and cocoa powder (291.7 …
mg/kg), iron (36.8 ± 1.10 mg/kg), zinc (6.36 ± 524.7 mg/kg) and, respectively, Mahmoud et
0.36 mg/kg) and fibres (16.50 g/100 g). al. (2012), in case of fenugreek supplemented
Moisture of Aronia fruits was 10.2%, and total biscuits (465.10 mg/kg d.w., respectively,
sugar content 30.25%. 561.7 mg/kg d.w.). Magnesium content of
Chemical composition of bakery products products varied in the range 839.20 ... 1196.6
fortified with Aronia is presented in Table 3. mg/kg, being comparable with those of the
It is noteworthy that these products have high achieved biscuits by Vitali et al. (2009) with
protein content (11.92 ... 12.95%), fibres (9.13 inulin added and one of the following raw
... 16.60%) and mineral elements, ash varying materials: soy flour, amaranth, carob, apple
between 1.51 ... 2.03%. fibre or oat fibre. The higher iron content was
Due to the high fibres content and low sugar recorded for Biscuits with Aronia and ginger
content (4.33 ... 5.79%), bakery products with (65.27 mg/kg, higher than those reported by
Aronia can be beneficial in diet of peoples with Vitali et al., 2009), and the lower, in case of
type 2 diabetes and obesity. Bread with Aronia (10.97 mg/kg).
697
Total polyphenol content 4%, 6%, 8% and 10% of grape by-products has
Fortification of bakery products with dried a significantly higher polyphenol content
fruits of Aronia melanocarpa had a positive (334.32 ... 613.77 mg GAE/100 g d.m.), in
effect on total polyphenol content and their comparison with bread assortments with Aronia
antioxidant properties. The biscuits present the (Mildner-Szkudlarz et al., 2011). These results
highest total polyphenol content: 263.22 mg were achieved mainly due to the high content
GAE/100g in case of Biscuits with Aronia and of polyphenols of grape by-products (5895 ±
ginger and 221.58 mg GAE/100 g, respectively, 150 mg GAE/100 g d.m.), in comparison with
in case of Biscuits with Aronia and cinnamon those of the dried fruits of Aronia, and the
(Figure 5). difference in composition of products.
Total polyphenol content is higher than those Bread with Aronia and Minibaguette with
reported by Mildner-Szkudlarz et al. (2013), in Aronia have a higher content of polyphenols
case of biscuits made from wheat flour and than those recorded for bread with adding of
addition of 10% white grape pomace (211 mg 15% amaranth flour (173 ± 9 mg GAE/100 g
GAE/100 g). Also, total polyphenol content of d.m.) and of 15% quinoa flour (188 ± 7 mg
biscuits with Aronia exceeds those of biscuits GAE/100 g d.m.), respectively, and increase of
supplemented with 10% germinated fenugreek dose of these pseudocereal flours, to 30%,
(Trigonella Foenum Graecum) seeds flour: determines an increase of total polyphenol
196.58 mg GAE /100 g (Mahmoud et al., content of the achieved breads: 261 ± 4 mg
2012). GAE/100 g d.m. in case of amaranth flour and
254 ± 10mg GAE/100 g d.m. in case of quinoa
flour (Chlopicka et al., 2012). Also, in case of
bakery with Aronia, there is an inversion
relationship between total polyphenol content
and colour parameter L*, demonstrating that
presence of these compounds in composition of
products determines dark colour of them
(Figure 6).
Figure 5. Total polyphenol content and antioxidant

capacity of bakery products fortified with Aronia
Bread with Aronia and Minibaguette with

Aronia had close values of total polyphenol
content (193.34 mg GAE/100 g, respectively,
197.91 mg GAE/100 g), but small in com-
parison with biscuits. This fact can be
explained by difference in composition (lower
percentage of powder obtained from dried
fruits of Aronia melanocarpa) and by higher Figure 6. Correlation between total polyphenols
value of moisture. and colour parameter L* in case of bakery
Total polyphenol content of those two bakery products fortified with Aronia
products is superior to those recorded in case of
bread prepared with 10% of grape pomace Antioxidant capacity
powder (89.43 mg GAE/100 g; Hayta et al., Due, mainly, to phenolic compounds contained
2014). Grape pomace presents high antioxidant in powder of Aronia, bakery products fortified
capacity, due to high content in phenolic with it have antioxidant capacity which varied
compounds, such as proanthocyanidins (Özkan in the range: 1.91 ... 3.42 mg Trolox
et al., 2004). Equivalents/g (minimum value was recorded
It should be noted that bread prepared with for Bread with Aronia, and the maximum one
sourdough mixed rye and four different levels: in case of Biscuits with Aronia and ginger).
698
Between total polyphenol content and 0.834; Biscuits with Aronia and ginger - 0.694
antioxidant capacity there is a directly ... 0.849.
proportional relationship. The obtained results Shelf-life relative high of bakery products
are consistent with those obtained by Zheng fortified with Aronia can be explained mainly
and Wang (2003), mentioning that content of by antioxidant and antibacterial potential of
polyphenols of fruits of Aronia was strongly fruits of Aronia melanocarpa. Bräunlich et al.
correlates with their antioxidant capacity. In (2013) have shown that Aronia melanocarpa
case of bakery products fortified with Aronia in extracts can inhibit bacterial growth of
this study, between total polyphenol content Escherichia coli and Bacillus cereus in vitro.
and antioxidant capacity there is a linear Also, Liepiņa et al. (2013) have shown that
correlation, R2 = 0.8126. Antioxidant capacity extracts from fruits of Aronia melanocarpa and
of biscuits with Aronia is higher than those wild rowan (Sorbus aucuparia L.) inhibited the
recorded in case of biscuits enriched with grape growth of Gram-negative bacterium
marc extract: 0.79 ± 0.045 µmol Trolox/g Pseudomonas aeruginosa but did not have
(Pasqualone et al., 2014). Antioxidant capacity influence on Escherichia coli. However, in case
of those two assortments bread with Aronia of bread and minibaguette, lactic acid bacteria
(7.64 ± 0.73 µmol Trolox/g for bread and 11.16 from sourdough used for fermentation and final
± 0.45 µmol Trolox/g, respectively, for proofing, act as a natural antibiotic, thereby
minibaguette) is superior to bread which increasing the shelf-life of these products.
contains 2.5 ... 7.5% grape seed and has
antioxidant capacity in the range 4.15 ... 6.28 CONCLUSIONS
µmol Trolox/g d.m. (Meral and Dogăn, 2013).
This study showed that dried fruits of Aronia
Microbiological analysis melanocarpa are an important source for
Based on microbiological, sensory and fortification of bakery products. Bakery
peroxide index analyses it was established products achieved with adding of powder of
shelf-life of products with Aronia, as follows: dried fruits of Aronia melanocarpa have a
Bread with Aronia - 7 days; Minibaguette with complex biochemical composition and
Aronia - 7 days; Biscuits with Aronia and antioxidant potential.
cinnamon - 72 days; Biscuits with Aronia and Bakery products fortified with dried fruits of
ginger - 72 days. Aronia melanocarpa have a high polyphenol
Microbiological analysis of bakery and pastry content (193.34 ... 263.22 mg GAE/100 g),
products fortified with Aronia, at the end of proteins (11.92 ... 12.95%), fibres (9.13 …
shelf-life is presented in Table 5. 16.60%), potassium (4784,60 … 10547
mg/kg), calcium (1100.2 ... 1691.4 mg/kg),
Table 5. Microbiological analysis of bakery products magnesium (839.20 ... 1196.6 mg/kg) and iron
fortified with Aronia (10.97 ... 65.27 mg/kg). Due to high content of
fibres and low content of sugars (4.33 ...
5.79%), bakery products with Aronia can be
included in diet of people with type 2 diabetes
and obesity. Also, noteworthy is that those two
assortments of biscuits with Aronia, contain
Microbiological indicators determined come ingredients with hypoglicemiant effect (fruits
under provisions of the Commission Regula- of Aronia melanocarpa, ginger and cinnamon)
tion (EC) No. 2073/2005 on microbiological could have beneficial effects on glycemic
criteria for foodstuffs. equilibrium of consumers with type 2 diabetes.
During shelf-life water activity varied Antioxidant capacity of bakery products
differentiated depending on product, as fortified with dried fruits of Aronia
follows: Bread with Aronia - 0.938 ... 0.945; melanocarpa varied in the range 1.91 to 3.42
Minibaguette with Aronia - 0.926 ... 0.938; mg Trolox Equivalents/g. Biscuits with Aronia
Biscuits with Aronia and cinnamon - 0.615 ... and ginger recorded the higher value of
699
antioxidant capacity and Bread with Aronia had Hayta M., Özuğur G., Etgü H., Şeker I.T., 2014. Effect
the minimum value. of grape (Vitis vinifera L.) pomace on the quality,
total phenolic content and anti-radical activity of
Products achieved with Aronia received bread. Journal of Food Processing and Preservation,
qualifying „very good" at sensory analysis, 38, 980-986.
recording scores in the range: 19.44-19.92. Due Horszwald A., Andlauer W., 2011. Characterisation of
to use of sourdough in composition, bread and bioactive compounds in berry juices by traditional
minibaguette have crumb elastic and dense, photometric and modern microplate methods. Journal
of Berry Research, 1, 189-199.
proper texture and, in the same time, pleasant Howard L.R., Prior R.L., Liyanage R., Lay J.O., 2012.
and intense flavour. Colour of products with Processing and storage effect on berry polyphenols:
Aronia was apreciated by expert panel challenges and implications for bioactive properties.
receiving 4 or 5 points after evaluation. Journal of Agricultural and Food Chemistry, 60,
Biscuits with Aronia and ginger had the darker 6678-6693.
Hudec J., Bakoš D., Mravec D., Kobida L., Burdova M.,
colour (L* = 29.6), Turianica I., Hlušek J., 2006. Content of phenolic
compounds and free polyamines in black chokeberry
ACKNOWLEDGEMENTS (Aronia melanocarpa) after application of polyamine
biosynthesis regulators. J Agric Food Chem, 54, 3625-
This study was achieved through Core 3628.
Jakobek L., Drenjančević M., Jukić V., Šeruga M., 2012.
Programme (PN 12 48), supported by the Phenolic acids, flavonols, anthocyanins and
Ministry of Education and Scientific Research, antiradical activity of ʻNeroʼ, ʻVikingʼ, ʻGaliciankaʼ
contract 48 N/2012, project no. PN 12 48 01 03. and wild chokeberries. Scientia Horticulturae, 147,
56-63.
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701
702
INTEGRATING SOIL PH MEASUREMENT INTO AN INTERNET

OF THINGS APPLICATION
Adrian ZĂRNESCU, Răzvan UNGURELU, Marius-Ionuţ MACOVEI,

Gaudenţiu VĂRZARU
Syswin Solutions SRL, Bucharest, 26 Biharia Street, 3 Floor, District 1, Bucharest, Romania
Abstract
The paper presents a device for soil pH determination on-site and remote transmission of the result. Soil pH is an
indicator of the soil quality because it affects plant growing. Normally, soil samples are taken from the field and tested
in-door in aqueous solutions using ISO 10390: 2005 specification. However, it was an increasing demand for on-site
measurements materialized in a plethora of stand-alone devices. Most of them offer a narrow range of pH values, e.g. 3
to 8, because the majority of plants has an optimum range between 5.5 and 7.5. The Internet of Things exigencies have
led to design a node of multiple sensors including soil pH measurement with an increased precision. Therefore, it was
used a sensor for hydrogen ion activity measurement based on glass membrane as a junction between the soil solution
and a reference solution. The probe was calibrated using pH known solutions and then tested on different soil samples -
all data being remotely collected. The results emphasized the capability of the device to measure the soil pH on-site and
to send it to remote.
Key words: environmental monitoring, internet of things, precision agriculture, soil pH, wireless communication.
INTRODUCTION from authorized laboratories which could be

several tens of kilometers reach a price that
Precision agriculture and Agriculture 4.0 farmers will not pay. It is an increased demand
concepts imply, among other things, more data for a competitive solution for continuously soil
to the farmer. These require a lot of sensors that pH monitoring from remote. Practical solutions
will generate information. One of the most have emerged on the market based on
important parameters is soil pH. As known, soil indicators, colorimeters or different types of
pH can change during the year due to rainfall, electrodes sensitive to H+ activity (Plotog et al.,
quality of the water used in irrigation, 2016), but the lack of these methods is that data
fertilizers, pesticides, root respiration and is available randomly and it is eventually
decomposition of organic matter by inserted manually in a database. The accuracy
microorganisms. Also, soil pH depends on of these cheap devices is poor.
temperature and moisture conditions and can As the Internet of Things (IoT) technology
vary to with the soil depth. Temperature emerges, all sensors are required to be
changes the chemical activity of the hydrogen automatically controlled from the distance and
ions (H+), so the measurements of pH must integrated in environmental monitoring
include the temperature correction to a standard systems. There are mentioned remote sensing
temperature of 25°C. The whole variation of a (RS) methods based on bare-soil images and
soil can vary up to a unit of pH within a year, spectroscopic reflectance of soil samples for
which is quite a lot, since, for example pH 6.4 soil survey, mapping, and quantitative soil-
is a slightly acidic soil, while pH 7.4 is a property characterization (Yufeng et al., 2011),
slightly alkaline one. Therefore, the one work claiming that for pH, the soil map
information must arrive in time to the farmer in and the RS estimate were nearly as accurate
order to take corrective measures. (Roelofsen et al., 2015). The methods, which
Currently, soil pH determination is done in imply satellite and aerial imagery or other non-
laboratories on samples of mixed soil in an contact nature of measurement, are costly.
aqueous solution; although very accurate, the There are mentioned real time and maped-
method is costly, takes time, and the distance based approaches using vehicle-based on-the-
703
go soil sensors for measuring soil properties the resolution +/- 0.0001 (Device #1). It has
including electromagnetic and even been chosen this type of sensor because the
electrochemical sensors for pH determination manufacturer states that the pH probe may have
(Adamchuk et al., 1999). Another work states in the soil indefinite placement. The life
that automated soil sampling system could be expectancy of the probe is more than 2.5 years
used to estimate soil pH on-the-go (Adamchuk without maintenance.
et al., 1999). The data can be sent to a remote The Signal conditioning block adapts the
center through the vehicle communication analog signal from the output of the pH sensor
station. The disadvantages of the method: the into a digital signal to be applied to any
values could not be as accurate as a laboratory microprocessor that supports UART, or I2C
test, the vehicle could not access any place and protocol.
the data is available discontinuously. The power supply block assures the voltages
In this context, the paper presents the way the for the electronic circuits from a solar cell
authors have solved the objectives they have panel and a rechargeable battery.
proposed: in-field soil pH determination with
incresed accuracy and the reception of data at a
remote center using wireless communication
technology.
The pH sensing is part of a more complex
device for monitoring environmental
parameters such as air (temperature, relative
humidity, harmful substance concentration,
wind speed and direction), soil (temperature,
relative humidity, pH, macronutrient content)
and lighting level. The goal was to build an
automatic system dedicated to coordinate and
streamline all resources and factors involved in
optimizing agricultural crops.
In order to meet the proposed objectives it was

designed an electronic module which interface
Figure 1. The block diagram of the electronic module for
a pH sensor to a microcontroller to read the in-filed pH measurement and data transmission to remote
data and to send it to remote by means of a
radio-frequency (RF) block (Figure 1). This On the reception side of the data it has to be
approach allowed to implement a complete IoT used a Wi-Fi gateway connected to a laptop to
application for agriculture. Since the main store and process the information.
requirement for an IoT application is the low The goal of the experiment was to collect the
power, the circuitry was designed around a low data regarding the soil pH from a remote center
power 8-bit AVR RISC-based microcontroller, in real time and to analyze the accuracy of the
ATmega2560, Microchip Technology. measurement.
Generally, the largest power consumption in an The soil pH determination shall be compared
IoT application is the RF block, therefore, it with the measurements made by other devices:
was selected the long-range low-power I) EC500, a pH - Conductivity meter from the
technology wireless platform that is the top EC510 kit, Extech Instruments/FLIR Systems,
technology choice for building IoT networks, which has a pH range between 0.00 to 14.00, as
LoRa. In addition, it uses a license-free benchmark ((Device #2);
spectrum. II) A cheap 3-in-1 moisture meter with light
The pH probe was a silver/silver chloride and pH test function for gardening, which offer
sensor, Atlas Scientific LLC, which can a pH range between 3.5 to 8, Shenzhen Xing
measure pH of solutions in the range 0-14 with Ying Da Industry Co (Device #3).
704
Before starting the experiments the pH sensor covering most of its scale: acidic, neutral and
was calibrated. The measurements were alkaline;
performed in the laboratory area on flower soil - three equal amount of mixed soil and aqueous
mixed with water at room temperature (25°C). solution were prepared and measured with all
According to the manufacturer’s three pH meters;
recommendations the measurement of the soil - it is supposed that samples equal amount of
pH should be preformed with a delay of 25 the same soil mixed with three different
minutes after the immersion in the mud. aqueous solutions change their properties
In the Figures 2 and 3 there are presented the regardless the measuring instrument;
pH devices used for comparision. - the recorded data from the Device #1 will be
analyzed and discussed.
The following three solutions were prepared:
alkaline (pH 13.2 on both devices, ALKALINE
indication on Device #3), approximately
neutral using running water (pH 7.82, pH 7.88
and slightly over 7 on Device #3) and acidic
(pH 4.5 on both devices, 7 on Device #3).
The solutions were poured into equal pots with
soil and the devices were infiltrated into the
created mud. The automatic reading period of
the pH values for the Device #1 was set to 10
minutes; at the same time the values of the
Figure 2. EC510 conductivity testing kit including: other devices were noticed by an operator.
EC500 meter, 3 calibration standards, 3 pH buffer
pouches plus rinse solution
The data from the pH sensor could be read on

the monitor of a laptop (Figure 4). The pH
sensor and the laptop were placed in two
different rooms of the company.
Figure 3. 3-in-1 plant flowers soil tester pH tester/

moisture meter/light meter
The experiments were performed in the

laboratory of the company; the temperature was
around 25°C ± 3°C.
Since the rezult of the measurements performed
with the Device #1 are available only on a Figure 4. An extract from the print screen on the laptop
remote display where the data can be stored monitor
and processed, while the other two devices
needed a local operator for readings, the This prove the main parts of the designed
following method was proposed: electronic module - microcontroller and LoRa
- three equal amount of aqueous solutions were transmission - work correctly.
prepared and measured with all three types of The interconnection between the computer and
pH meters; each solution had a different pH the pH sensor device offer the possibility to
705
transmit several commands: the period of For both very strongly alkaline solution and the
continuous readings, the single reading mode, moderately acidic solution the measurements
start/stop of readings in order to save power, have shown that the soil pH has decreasing
mathematical compensation of pH readings values. The Device #3 indications were
with temperature. ALKALINE for the first case and 7 for acidic.
The results for neutral domain are displaying a A graphical representation of the cumulative
slow variation for Devices #1 and #2, while measurements using the Device #1 is presented
Device #3 shows always a value between 7 and in Figure 6. The measurements were performed
8 (Table 1). during a period of more than 3.5 hours. The
slowly decreasing trend of the measurements is
Table 1. Determination of soil pH with three different due to the gravitational leakage of the water
devices for neutral domain from the created solution into the soil. This is a
Soil pH
situation that is also encountered in practice.
Time [min.]
Device #1 Device #2 Device #3
0 6.939 7.27 7-8
10 6.960 7.20 7-8
20 6.958 7.22 7-8
30 6.945 7.14 7-8
40 6.942 7.16 7-8
50 6.958 7.08 7-8
60 6.864 7.07 7-8
70 6.849 7.01 7-8
80 6.850 7.03 7-8
90 6.850 7.02 7-8
100 6.855 7.01 7-8
110 6.854 7.12 7-8
Their graphical representation is presented in

Figure 5. The measurements performed with
the EC500 meter have a greater margin of Figure 6. Graphical representation of soil pH cumulative
variation when it is immersed in the mud. The measurements for Device #1
difference between the maximum and the
minimum values of soil pH is 0.260 while for It is supposed that during the laboratory tests
the Device #1 is 0.111. However, this is an the pH values will tend to fixed value due to
improper use of the probe it has to be used only the limitation of the container with the soil.
for aqueous solutions. In order to test the repeatability of
measurements it was recorded the variation of
the pH values received from the Device #1
during a longer period. As it can be seen in the
Figure 7, during 9 hours the soil pH values
were practically constant. It is a real situation
because the solution was almost neutral and
during the period of test the conditions in the
laboratory were unchanged and no other
material was added.
The lack of extremely values proves the high
quality of the pH sensor.
Although it is supposed that the pH sensor will
stay in the soil much longer time than tested
above, in order to make correct determinations
Figure 5. Graphical representation of soil pH it will be created around it a mixed solution
measurement for near neutral domain
using pure water (pH 7) every time when
desired.
706
Figure 7. The graphical representation of pH measurement performed with Device #1 during 9 hours
The practical implementation of the pH standard pH instrument for different aqueous

measurement and the remote transmission of solutions. The extrapolation of the measure-
the data is presented in Figure 8. ments to the soil mixed with water has to be
confirmed by a specialized laboratory.
The pH sensor device could be programmed to
enter single reading mode, standby mode and
multiple readings with different periods of
readings.
The measurements using inexpensive 3-in-1
device did not prove to be realistic.
The designed pH device can be interconnected
to other systems for parameter monitoring in
agriculture that have UART or I2C protocol
capability.
ACKNOWLEDGEMENTS
This research work was carried out in the frame

of the „Operational Program Competitivity
2014-2020” POC - A.1 – A.1.2.1. – D – 2015,
Figure 8. View of the multi sensors device with COD SMIS 104238, supported by MECS -
autonomous energy and wireless communication ANCSI, PRIAMM, ID p_39_360 project.
including soil pH measurement
REFERENCES
CONCLUSIONS
Adamchuk I.V., Morgan M., 1999. Evaluation of
automated soil pH mapping. Agricultural and
In-field soil pH determination and remote Biological Engineering, Purdue University. Paper No.
transmission of the values in real time was 991100.
proved. Adamchuk I.V., Paul J., 2002 On-the-go vehicle-based
The data can be stored and processed on a soil sensors. Precision Agriculture, University of
Nebraska Cooperative Extension, EC 02-178.
computer placed anywhere in the cloud. Plotog I., Vărzaru G., Iacomi C., Iacomi B., Roşca I.,
The numerical values of the pH measured by 2016. Investigations on a practical determination of
the proposed device were similar compared to a soil pH for an irrigation system. Scientific Papers, vol.
707
59/2016, Series Agronomy, University of Agricultural Applied Earth Observation and Geoinformation,
Sciences and Veterinary Medicine Iaşi. Volume 43, December 2015, Elsevier, pp. 149-159.
Roelofsen H.D., Peter M. van Bodegom, Lammert Yufeng Ge, J. Alaex Thomasson, Ruixiu, 2011. Remote
Kooistra, Jorg J. van Amerongen, Jan-Philip M. Witte, sensing of soil properties in precision agriculture: A
2015. An evaluation of remote sensing derived soil review. Frontiers of Earth Science, Volume 5, Issue 3,
pH and average spring groundwater table for Springer Link, 229-238.
ecological assessments. International Journal of
708
ADVANTAGES AND DISADVANTAGES OF PESTICIDE ANALYSIS

METHODS USED IN AGRICULTURAL SAMPLES
Elena-Cornelia MITRAN1, 2, Elena Perdum1, Ovidiu George IORDACHE1,

Iuliana DUMITRESCU1
1
National Research and Development Institute for Textiles and Leather Bucharest (INCDTP),
16 Lucrețiu Pătrăşcanu Street, 030508, Bucharest, Romania
2
Politehnica University of Bucharest, 1-7 Polizu Street, Bucharest, Romania
Abstract
Pesticides are substances (herbicides, fungicides, insecticides, plant growth regulators etc.) used primarily for pest
control that can occur in both animals and plants. Unfortunately, besides beneficial effects, their use also has many
disadvantages, these being toxic to humans and environment. For this reason, it is very important to have precise and
accurate analytical methods for pesticide determination and quantification. The main purpose of this paper was to
provide a description of the most commonly used methods of analysis and sample preparation for qualitative and
quantitative determination of pesticides. The field of agriculture was analysed as a field of use of pesticides. Thus it has
been highlighted that current analysis methods heavily rely on the use of gas chromatography analysis tools and in
regard to the methods of agricultural samples preparation, these are in general extraction methods. The paper presents
the advantages and disadvantages of the mentioned methods, in qualitative and quantitative assessment of pesticide
present in agricultural biomass.
Key words: pesticides, agriculture, analytical method, chromatography.
INTRODUCTION 2015), genotoxic effects in hamster ovarian

cells (Soloneski et al., 2015) and last but not
Pesticides are substances used primarily for least increased risk for dementia (Lin et al.,
pest control (US Environmental, 2007) that can 2016).
occur in both animals and plants. There are three ways in which humans can be
There are several classes of pesticides, the most affected by pesticides (Yusà, Coscollà and
common ones being: Millet, 2014) and the most important source is
- Organochlorine pesticides: DDT is the most through diet or ingestion.
used pesticide from this class and although its Another way is through dermal contact, a way
use has been restricted, there are countries that that it’s more and more encountered due to
are thinking of reintroducing it (Turusov et al., household use of pesticides.
2002; Van den Berg, 2009); And a third way is by inhalation of
- Organophosphorus pesticides: although they contaminated air, particularly for those are
are said to be a more ecological option, many staying nearby the agriculture areas.
of the substances from this class have been Pesticides besides activities that have a
associated with an endocrine disrupting negative impact on the health of people, affect
potential (Mnif et al., 2011; Karami-Mohajeri on long term all type of water (surface and
et al., 2011); underground), air, soil and also soil organisms
- Carbamate pesticides: unfortunately the use of (Sarfraz et al., 2009)
this class of pesticide can lead to a series of To understand better the importance of
negative activity on the human body such as: pesticide use a short history of these
possible reproductive disorders (Jamal et al., compounds have been summarised in Table 1.
709
Table 1. Brief history of pesticide use (Jojiya et al., 2017)
Period Type of pesticide used

Ancient time Ashes, common salts and bitters
Arsenic, suggestion of soda and olive oil for treatment of legumes (Pliny the Elder, a
1st century
Roman naturalist-Historia naturalis)
16th century Arsenicals and nicotine in the form of tobacco extracts (Chinese farmers)
1850 Pyrethrum, soap and a wash of tobacco, sulfur and lime also used
The pigment Paris green (impure form of copper arsenite), Paris green and kerosene oil
1867
emulsion
1896 Bordeaux mixture (CuSO4 and Ca(OH)2) and selective chemical herbicides
1900 Dilute sulfuric acid, copper nitrates and potassium salts
Sodium arsenite solutions become the standard herbicides and are used in large
1900-1950
quantities
1913 Organomercury seed dressing
1913-1939 Dithiocarbamates fungicides used in US
Insecticidal potential of DDT discovered in Switzerland. Chlorinated hydrocarbons
1939
(DDT, BHC, dieldrin, aldrin and chlordane)
1950s Fungicides captan, glyodin and organophosphorus insecticide: malathion
1961 DDT registered for use on 34 different crops as pesticide usage dramatically increases
Bio accumulation and long-term toxicity and pest resistance became evident. Stoppage
1962 of DDT usage and other chlorinated compounds by farmers. Favor of the use of
Organophosphates and Carbamates
Environmental Protection Agency revoked the use of DDT on all food sources in the
1972 United States. The World Health Organization, however, still reserves the right to use
DDT on particularly virulent outbreaks of malaria
Herbicidal sulfonylureas, neonicotinoids, glyphosate, synthetic fungicides such as
1972-1980
metaxyl and triadimefron and light-stable pyrethroid pesticides are introduced
Integrated pest management, intensified research on biological pest control methods
1990s
and other alternatives to pesticides
1990-1995 Increased interest in Integrated Pest Management (IPM) programs
Wide spread usage of IPM techniques organic farming excluding the usage of synthetic
2000
pesticides.
Involvement of genetic engineering and biotechnological methods to control the usage
2010-2015
of pesticides eg. baculoviruses
EXTRACTION METHODS OF has several advantages such as: environ-

PESTICIDES mentally friendly and is simple and fast.
Another type known and used extraction
A novel method for sample extraction that it is method is Supercritical Fluid Extraction (SFE)
used more and more is QuEChERS (Sherish et which can be used for both solid and semi-solid
al., 2017). samples. It is a recently developed method. The
One study use QuEChERS for the main advantage is that SFE is simple and less
determination of seven pesticides from time consuming. Teresa Castelo-Grande et al.
Okavango Delta water samples and present a developed a SFE method for pesticides from
detection limit situated between 0.102 μg/L- soil and obtain a recovery of atrazine higher
1.693 μg/L and a recovery value situated that 96% (Teresa Catselo-Grande et al., 2005).
between 61% and 95% (V.C. Obuseng et al., Tatsuo Yoshida et al. obtain a value for
2013). Correia-Sa et al. have analysed soil recovery of Isotianil extracted from rice and
samples with organic carbon over 2.3% and rice cultivation soil between 95.1% and 99.3%
obtained 3.42-23.77 μg/kg limit detection and (T. Yoshida et al., 2013).
70%-120% recovery (Correia-Sa et al., 2012). Hiroaki Chikushi et al. (H. Chikushi et al.,
Fresh peppermint samples were analysed by 2009) evaluated the presence in water sample
Magdalena Slowik-Borowiec et al. using this and proposed a method with low limit of
type of extraction method and obtain 0.01 detection: 0.002-2.3 µg/l.
mg/kg limit of detection and 100% recovery A quite new extraction method is accelerated
(Slowick-Borowiec et al., 2012). This method solvent extraction (ASE). Important points in
710
this technique are temperature and pressure and and 0.6-3.1 µg/kg limit of detection (Michelle
also have advantages like speed and simplicity. L. Hladik et al., 2012).
Another plus is represented by the volume of Beside the extraction methods mentioned
reactive which is relatively low. Michelle L. above, there are presented in Table 2 other data
Hladik et al. have analysed environmental obtained by using different types of extractions.
sediment samples and obtain 81-101% recovery
Table 2. Extraction method of pesticide
Extraction Recovery/Limit of
Sample Advantages/Disadvantages Reference
method detection (LOD)
Advantages: Possibility of using
Headspace
LOD = 0.07-12.54 various solvents; Very good for
Single-Drop Amvrazi et
µg/kg Honey extraction of diazinon; Possibility of
Micro-extraction al., 2012
Recovery = 74-102% extracting volatile and water-soluble
(HS-SDME)
analytes.
LOD = 0.01-0.088 Advantages: Can be used to
Lopez-
Solid Phase µg/L determine may types of pesticides;
Water samples Mesas et al.,
Extraction (SPE) Recovery = 74.2- Present very good limit of detection;
2007
116.4% Rapid and efficient method.
Advantages: Fast; Sensitive; Multi-
Dispersive
LOD = 0.0032- residue method; Very good recovery.
liquid–liquid Abdullash et
0.0174 µg/L Water samples Disadvantages: Limited solvent
micro-extraction al., 2017
Recovery = 84-108 % choice; It’s not suitable when the
(DLLME)
matrix composition is complex.
Solvent-based de- Advantages: Environmentally
emulsification friendly; Less expensive than other
dispersive liquid– techniques; Can be applied also for Caldas et
Recovery = 60-120% Water samples
liquid micro- pharmaceuticals and personal care al., 2016
extraction (SD- products.
DLLME) Disadvantages: same as DLLME.
Sedimentation of
LOD = 0.6-3.1 µg/kg Advantage: Small volumes of
Accelerated agricultural
Recovery = 81-101% solvents; Fast, easy and simple. Hladik et
solvent extraction drainage samples
Disadvantages: Relatively high price al., 2012
(ASE) LOD = 0.8-3.4 µg/kg Sediment samples of the equipment.
Recovery = 75-102% from the estuary
Solid-phase
extraction (D- Advantages: Alternative materials are
SPE) by quick, more effective and less expensive
Arias et al.,
easy, cheap, LOQ = 0.1-100 µg/kg Rice paddy soils than traditional sorbents
2014
effective, rugged Disadvantages: Limited solvent
and safe choice for extraction.
(QuEChERS)
Cereals (corn, Advantages: Multi-residue analysis;
He et al.,
QuEChERS Recovery = 70-120% wheat flour and Simple and with satisfactory
2015
rice) accuracy.
Advantages: Can be applied on all
Direct immersion
types of water samples;
solid-phase LOD = 0.015-0.13 Tankiewicz
Aqueous samples Multi-residue analysis.
micro-extraction µg/L et al., 2013
Disadvantages: Relatively expensive
(DI-SPME)
because of fiber cost; Matrix effects.
Microwave-
Recovery = 81.5-
assisted Grass samples;
108.4% Both methods are suitable for
extraction (MAE) Vegetation from Barriada-
chlorinated pesticides analysis.
the contaminated Pereira et
Advantages MAE: shorter extraction
Soxhlet Recovery = 75.5- industrial area of al., 2003
times, higher extraction rates.
extraction 132.7% Torneiros
711
DETERMINATION METHODS OF The real samples have included city piped and
PESTICIDES agricultural water.
Another determination method that uses
Javad Ghodsi and Amir Abbas Rafati have HPLC/MS/MS was developed by Hwa-mi Lee
developed a method for the determination of et al. They analyse 56 residual pesticides from
diazinon made by use of a MWCNTs/TiO2NPs commercial crops and obtain a recovery value
nanocomposite sensor (J. Gjodsi et al., 2017). between 65-82% and a detection limit up to
Comparative with other methods used such as 11.54 µg/kg (Hwa-mi Lee et al., 2013).
gas chromatography, high-performance liquid Hirahara et al. described a screening method
chromatography (HPLC), mass spectrometry for determination of 200 pesticides using
method, spectrophotometry, infrared spectro- GC/MS/MS.
scopy and an enzyme immunoassay (M. The method present a recovery value situated in
Khadem et al., 2017; T.D. Lazarevic-Pasti et 50-150% interval and a good limit of
al., 2013; G. Erdogdu, 2003) that are ex- quantification of 0.01 mg/kg (Hirahara et al.,
pensive, time-consuming and with the need of 2006).
trained employers, this method has shown to be There are many methods for determination of
sensitive, fast and use an easy fabricate sensor pesticides, some of which are presented in the
that is not so expensive. Also the method Table 3.
presents a good limit of detection of 3 nM.
Table 3. Determination method of pesticide
Determination Recovery/ Limit of
Sample Advantages/Disadvantages Reference
method detection (LOD)
Very good for determination
of organochlorines and
Amvrazi et
GC-ìECD LOD = 0.07-19 µg/kg Honey organophosphorus pesticides;
al., 2012
Highly sensitive;
Low detection limit.
Sedimentation of
LOD = 0.6-3.1 µg/kg
agricultural
Recovery = 81-101%
drainage samples Very good recovery value; Hladik et
GC/MS
Sensitive method. al., 2012
LOD = 0.8-3.4 µg/kg Sediment samples
Recovery = 75-102% from the estuary
Micellar electro-
Advantages:
kinetic
LOD = 0.001-0.144 Environmental or Low limit of detection Moreno-
chromatography–
µg/L drinking water Disadvantages: Gonzalez et
electrospray–mass
Recovery = 83-101% samples Relatively high price of the al., 2012
spectrometry
equipment
(MEKC-ESI-MS/MS)
Ultra-high-
performance liquid Multi-residue method;
chromatography/time- Vegetable and fruit suitable for routine Sicaperumal
Recovery = 74-111%
of-flight mass samples quantitative analyses of et al., 2015
spectrometry pesticide
(UHPLC/TOF-MS)
LOD = 0.4-48.2 µg/kg Advantages:
GC/SQ-MS
Recovery = 70-110% The method is repeatable;
Can be used in many types of
Grape, lemon, Lesueur et
matrices
LOD = 1-115 µg/kg onion and tomatoes al., 2008
HPLC/IT-MS Disadvantages:
Recovery = 70-110%
Lemon and onion showed
poor recoveries
Cereal samples
Improved analytical
Recovery = 96 ± 9% (wheat, rye, barley, Walorczyk
GC-MS/MS performance parameters
oats, maze, et al., 2012
Multi-residue method.
buckwheat)
712
CONCLUSIONS Microwave-assisted extraction versus Soxhlet
extraction in the analysis of 21 organochlorine
pesticides in plants, Journal of Chromatography A,
Due to the numerous negative effects on human 1008 (1): 115-122.
and environmental, it is important and there is Caldas S.S., Rombaldi C., Arias J.L.O., Marube L.C.,
still a need to develop precise, sensible and Primel E.G., 2016. Multi-residue method for
robust extraction and analysis methods to determination of 58 pesticides, pharmaceuticals and
determine the amount of pesticides and to keep personal care products in water using solvent
demulsification dispersive liquid-liquid micro-
them in conformity with applicable laws. extraction combined with liquid chromatography-
Taking into account all these aspects, in this tandem mass spectrometry, Talanta, 146: 676-688.
paper we have briefly discussed the most Castelo-Grande T., Augusto P.A., Barbosa D., 2005.
commonly used extraction and determination Removal of pesticides from soil by supercritical
method for pesticides mainly from agriculture extraction - a preliminary study, Chemical
Engineering Journal, 111: 167-171.
domain and also from other fields. Chikushi H., Hirota K., Yoshida N., Edamura T., Toda
K., 2009. Simple water analysis of golf link pesticides
ACKNOWLEDGEMENTS by means of batch-wise adsorption and supercritical
fluid extraction, Talanta, 80 (2): 738-743.
This work was carried out in Nucleu Correia-Sa L., Fernandes V.C., Carvalho M., Calhau C.,
Domingues V.F., Delerue-Matos C., 2012.
Programme TEX-PEL-2020, implemented with Optimization of QuEChERs Method for the Analysis
the support of MCI, project nr. PN 18 23 01 02, of Organochlorine Pesticides in Soils with Diverse
entitled „Exploitation of filamentous fungi for Organic Matter, Journal of Separation Science, 35
the production of bio-composite materials”. (12): 1521-1530.
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the determination of diazinon insecticide, Journal of
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Ghodsi J., Rafati A.A., 2017. A voltammetric sensor for
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TiO2 nanoparticles covered multi walled carbon
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Hirahara Y., Kimura M., Inoue T., Uchikawa S., Otani
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He Z., Wang L., Peng Y., Luo M., Wang W., Liu X.,
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714
SCREENING OF MICROORGANISMS DISPLAYING ACETYL XYLAN

ESTERASE ACTIVITY
Aglaia BURLACU, Florentina ISRAEL-ROMING, Călina Petruța CORNEA

Abstract
Hemicellulose is a major component of the lignocellulosic biomass, located between lignin and cellulose fibers and is
the second most abundant natural polymer on earth. The main constituent of hemicellulose is xylan, a polysaccharide
comprised of β-1.4-linked xylopyranosyl residues. Due to its complex structure and heterogeneous nature, hydrolysis of
xylan requires a synergic action of several enzymes generally named xylanases. Amongst them, acetyl xylan esterase
(AXE) is an accessory enzyme that liberates acetyl groups from the side chains of the xylan backbone. These acetyl
side-groups protect the backbone of xylan from the action of others enzymes, therefore their release facilitates the
action of endoxylanases. Several microorganisms display acetyl xylan esterase activity, the main producers being fungi
and bacteria. The aim of this study was to test different strains in regard to their ability to display acetyl xylan esterase
activity. The microorganisms subjected to screening belonged to the genres: Aspergillus, Penicillium, Bacillus,
Trichoderma and Fusarium. The strains subjected for screening were also studied for their xylanase activity, in order to
compare their ability to produce both xylanase and acetyl xylan esterase. In addition, the microorganisms selected after
the screening process were subjected to protein assay, in order to determine the specific enzymatic activities. The best
acetyl xylan esterase activities were detected with Aspergillus brasiliensis ATCC 16404 UV 7, Aspergillus brasiliensis
ATCC 16404 UV 5, Aspergillus niger UV 10 and Penicillium digitatum UV 11. These experimental results are
significant for further studies related to hydrolysis of hemicellulose, regarding lignocellulosic biomass valorisation.
Key words: acetyl, xylan, esterase, xylanase.
INTRODUCTION efficient enzymatic hydrolysis of lingo-

cellulosic materials (Zhang et al., 2011).
Lignocellulosic materials, such as forestry and Acetyl xylan esterases (AXE, E.C. 3.1.1.72) are
agricultural wastes, are the major source for key accessory enzymes involved in deacety-
renewable organic matter (Pothiraj et al., 2006; lation of xylans and xylo-oligosachharides.
Yoo et al., 2014). They are able to hydrolyse acetyl groups from
Lignocellulose has three main components: D-xylopyranosyl residues in xylan chains (Hou,
cellulose, hemicellulose and lignin, with 2005).
hemicellulose as the second most abundant After the hydrolysis of the acetyl ester groups,
natural heteropolymer on earth (Hendriks and xylan main chain has new ester-free regions
Zeeman, 2009). targeted by β-1.4-endoxylanase. Acetylxylan
Xylan, as the main component of hemicel- hydrolysis by endoxylanases proceeds with a
lulose, consists of β-1.4-linked xylopyranosyl higher and faster rate when AXEs are involved.
residues (Blum et al., 1999). (Hou, 2005; Zhang et al., 2011)
It can be degraded by the synergic action of Due to the lack of low-cost and well
several enzymes such as: β-1.4-endoxylanase, characterized acetylated xylo-oligosaccharides
β-xylosidase, α-L-arabinofuranosidase, α-glu- and xylans, screening of microorganisms
curonidase, acetyl xylan esterase and phenolic displaying AXE activity is usually conducted
acid (ferulic and p-coumaric acid) esterase by using chromogenic or fluorogenic acetyl-
(Dhiman et al., 2008; Motta et al., 2013). esterase substrates such as α-naphtyl acetate, β-
Several reports suggest that enzymatic removal naphtyl acetate, 4-nitrophenyl acetate or 4-
of xylan enhances cellulose hydrolysis, methylumbelliferyl acetate (Biely et al., 2014;
therefore it is important to improve xylan Johnson et al., 1988; Shao and Wiegel, 1995).
enzymatic degradation in order to obtain an In addition, some methods use as a substrate
715
chemically acetylated xylan, obtained usually UV 5 and A. brasiliensis UV 14), 40 minutes
by Johnson’s method (1988) (Hou, 2005). (A. brasiliensis UV 7 and P. digitatum UV 6),
Some studies have concluded that 50 minutes (A. niger UV 10 and Penicillium
supplementation of culture medium with digitatum UV 11) and 60 minutes (A. niger UV
Tween 80 could improve significantly AXE 20, A. brasiliensis UV 5 and Penicillium
and xylanase activities, due to the fact that digitatum UV 12).
Tween 80 has a strong impact on the
microorganism ability to synthesize the Primary screening of microbial strains for
enzymes (Atta et al., 2011). AXE activity
There are several AXE producing microor- The selected isolates were cultivated on two
ganisms studied over the years, including fungi different liquid mediums containing 0.8%
and bacteria: Penicillium purpurogenum, P. corncob xylan as the only carbon source; only
notatum, P. chrysogenum, Phanerochaete one medium supplemented with 0.5% Tween
chrysosporium, Schizophylum commune, 80. Other components of the cultivation
Aspergillus awamori, A. niger, Bacillus medium were for fungi (g/L): 0.05 g
pumilus, B. subtilis, Thermoanarobacterium MgSO4·7H2O, 0.005 g CaCl2, 0.005 g NaNO3,
sp., Streptomyces lividans, S. flavogriseus, 0.009 g FeSO4·7H2O, 0.002 g ZnSO4, 0.012 g
Fusarium oxysporum, Trichoderma reesei, T. MnSO4, 0.23 g KCl, 0.23 g KH2PO4, 2 g
longibrachiatum, Aureobasidium pullulans etc. peptone (Adesina and Onilude, 2013) and for
(Atta et al., 2011; Bajpai, 1997; Biely et al., bacteria (g/L): 0.05 g MgSO4·7H2O, 0.05 g
1988; Degrassi et al., 1998; Halgasová et al., NaCl, 0.01 g CaCl2, 0.2 g yeast extract, 0.5 g
1994; Hou, 2005; Yang et al., 2017). peptone (Mahilrajan et al., 2012).
The aim of this study was to test different The Erlenmeyer flasks containing the
microbial strains regarding their ability to cultivation medium were inoculated with the
display acetyl xylan esterase activity, under selected microorganism and incubated at 30 ±
different cultivation conditions. 2°C in an incubator with shaker at 140 rpm for
5-9 days depending on the strain. Samples were
MATERIALS AND METHODS taken at every 24 hours and centrifuged at 4500
rpm for 30 minutes. The supernatants were
Microorganisms subjected to acetyl xylan esterase assay.
The bacterial and fungal isolates studied were
provided by the Department of Genetics and Secondary screening
genetic engineering of the Faculty of The isolates that displayed high acetyl xylan
Biotechnologies from USAMV Bucharest. esterase activity were subjected to a second
For this study, several microbial isolates were screening being cultivated on the same liquid
subjected to screening: wild strains (Bacillus medium, but with different carbon sources:
amyloliquefaciens B4, B. amyloliquefaciens wheat bran (WB) and de-starched wheat bran
BN7, Trichoderma viride TV2, Fusarium (DSWB), supplemented with 0.5% Tween 80.
oxysporum, Aspergillus niger, A. brasiliensis De-starched wheat bran was prepared
ATCC 16404, Penicillium digitatum) and according to Mukherjee (2007) and Huang
mutant strains (A. niger UV 5, A. niger UV 10, (2013). Samples were taken at every 24 hours
A. niger UV 20, A. brasiliensis UV 5, A. and were analysed for their acetyl xylan
brasiliensis UV 7, A. brasiliensis UV 14, esterase activity and xylanase activity.
Penicillium digitatum UV 6, Penicillium
digitatum UV 11, Penicillium digitatum UV Acetyl xylan esterase assay
12). The enzymatic activity of AXE was measured
The mutant strains were obtained after random by hydrolysis of p-nitrophenyl acetate (pNPA)
mutagenesis through UV irradiation according to p-nitrophenol (pNP), according to a modified
to Ho and Ho method (2015). Briefly, the spore method by Johnson et al. (1988) (Atta et al,
suspensions were kept at a distance of 10 cm at 2011). The assay mixture consisted of 1mL
254 nm in a vertical laminar flow cabinet for 0.1M sodium phosphate buffer (pH 7), 0.9 mL
different time exposure: 30 minutes (A. niger 10 mM pNPA and 0.1 mL enzyme sample and
716
was incubated at 37ºC for 10 minutes. The conditions, lower than the results of other
release of pNP was measured by reading the studies (Atta et al., 2011).
absorbance at 410 nm using a spectropho- The best AXE activity were recorded with
tometer. One unit of acetyl xylan esterase fungal strains (Table 1), such as: A. niger UV
activity was defined as the amount of enzyme 10, A. brasiliensis UV 5, A. brasiliensis UV 7
required to release 1 µmol of p-nitrophenol per and P. digitatum UV 11. Amongst them, A.
minute under the specified assay conditions. brasiliensis UV 7 had the best AXE activity on
corncob xylan medium (2.68 µmol/ml/min.),
Xylanase assay while A. brasiliensis UV 5 had the highest
Xylanase activity was determined according to activity of 3.24 µmol/ml/min. on corncob xylan
the DNS (3.5-dinitrosalicylic acid) assay for medium supplemented with Tween 80,
reducing sugars (Bailey et al., 1992). Assay comparable with other studies (Christov et al.,
mixture consisted of 0.5 mL sample and 0.5 1993; Khan et al., 1990). The results obtained
mL of 0.6% oat spelt xylan and was incubated with the mutant strains were comparable or
at 40ºC for 10 min. The reaction was slightly higher than the activities recorded with
terminated by adding 1 mL of DNS and heating the wild type fungal strains (Table 1).
for 5 min at 80ºC. 3 mL of distilled water was Although other studies (Christov et al., 1993;
added to the mixture and after 30 min the Christakopoulos et al., 1999) suggest that
absorbance was read at 540 nm using a Fusarium oxysporum is an important producer
spectrophotometer to determine de amount of for AXEs, under this experimental conditions,
sugar released by the enzyme. One unit of the strain displayed a low AXE activity on both
xylanase was defined as the amount of enzyme cultivation mediums.
that released 1 μmol reducing sugar as xylose
equivalent per minute in the reaction mixture Table 1. Acetyl xylan esterase activity on xylan medium
under the specified assay conditions. with or without Tween 80
AXE activity
Protein assay (µmol/ml/min)
Microorganism Xylan +
The samples from the secondary screening Xylan
Tween 80
were analysed for their soluble protein medium
medium
quantification using Lowry method (1951). B. amyloliquefaciens B4 1.01 1.21
This analysis was necessary to determine the B. amyloliquefaciens BN7 0.29 0.33
specific enzymatic activity. T. viride TV2 1.40 1.67
Fusarium oxysporum 0.32 0.41
RESULTS AND DISCUSSIONS A. niger - wild type 1.78 2.19
A. niger UV 5 1.71 2.14
A. niger UV 10 2.31 2.93
Primary screening
A. niger UV 20 1.68 1.99
The wild and mutant strains obtained with
A. brasiliensis ATCC 1.81 2.15
random mutagenesis through UV exposure 16404 - wild type
were evaluated for their ability to produce A. brasiliensis UV 5 2.50 3.24
acetyl xylan esterases on xylan medium with or A. brasiliensis UV 7 2.68 3.19
without Tween 80. A. brasiliensis UV 14 1.92 2.37
From the bacterial strains, B. amyoliquefaciens P. digitatum - wild type 2.05 2.66
B4 was detected with the highest AXE activity P. digitatum UV 6 2.02 2.59
of 1.20 µmol/ml/min. on xylan medium and P. digitatum UV 11 2.34 3.03
P. digitatum UV 12 1.92 2.25
1.29 on xylan medium supplemented with
Tween 80 (Table 1). The results were higher
than other data regarding AXE activity of For P. digitatum mutant strains (Table 1), the
Bacillus sp. (Christov et al., 1993). AXE activities were comparable with the wild
As shown in Table 1, for the bacterial strains, strain, except for P. digitatum UV 11, that
the addition of 0.5% Tween 80 had a low displayed a relatively high AXE activity (3.03
improvement on AXE production (7.5% and µmol/ml/min) on xylan medium supplemented
13.8%) under this study experimental with Tween 80. This can suggest that UV
mutagenesis could be an important method for
717
improvement of microbial strains, regarding wheat bran medium (3.24 µmol/ml/min) and
their AXE activities. de-starched wheat bran medium (3.52
According to these results (Table 1), the µmol/ml/min), higher than the activities of the
addition of Tween 80 improved the enzymatic wild type strains with cca 29% (Table 2).
activity of the isolates with 20-30%, compa- In addition, the use of de-starched wheat bran
rable with other similar studies (Atta et al., instead of wheat bran as the only carbon source
2011). was correlated with an increase of enzymatic
activities of 7-10% (Table 2).
Secondary screening Regarding P. digitatum, AXE activities of the
After the first screening, several fungal isolates mutant isolates were higher with almost 16.5%
detected with the highest AXE activity were than the wild strain, while xylanase activities
subjected to a second screening in order to were higher with cca 6% than the wild type
examine the influence of the selected carbon strain (Table 2).
source upon AXE and xylanase activities.
Protein assay
Table 2. AXE and xylanase activity on wheat bran (WB) After the secondary screening, the samples
and de-starched wheat bran (DSWB) from the strains cultivated on de-starched
Microorganism AXE activity Xylanase activity wheat bran with Tween 80 were also analysed
(µmol/ml/min) (µmol/ml/min) for their protein content in order to determine
WB DSWB WB DSWB their specific AXE activity.
B. 1.05 1.14 1.72 1.89
amyloliquefaciens
The highest specific enzymatic activity of 1.34
B4 µmol/mg protein was recorded with A.
A. niger - wild type 2.10 2.31 2.44 2.67 brasiliensis UV 5 (Table 3), higher than the
A. niger UV 10 2.72 2.99 2.68 2.90 wild type strain with cca 12%.
A. brasiliensis 2.08 2.27 2.51 2.73
ATCC 16404 - wild Table 3. Specific AXE activity
type
A. brasiliensis UV 5 3.09 3.41 3.24 3.52 Microorganism Specific enzymatic
A. brasiliensis UV 7 3.10 3.35 2.87 3.1 activity (µmol/mg
P. digitatum - wild 2.48 2.72 2.63 2.89 protein)
type B. amyloliquefaciens B4 1.05
P. digitatum UV 11 2.89 3.17 2.80 3.05 A. niger - wild type 1.22
A. niger UV 10 1.43
The best AXE activity was detected with A.
brasiliensis UV 7 on wheat bran medium (3.10 A. brasiliensis ATCC 16404 1.20
- wild strain
µmol/ml/min.) and with A. brasiliensis UV 5 on
de-starched wheat bran medium (3.41 A. brasiliensis UV 5 1.34
µmol/ml/min.). AXE activities of the mutant A. brasiliensis UV 7 1.27
strains were higher than the activities of the
P. digitatum - wild type 0.78
wild type strains with almost 50% (Table 2).
Comparing the results of AXE activity from P. digitatum UV 11 0.81
table 1 and table 2, it can be observed that
when using wheat bran the microbial strains P. digitatum had a low specific enzymatic
show a slightly lower activities compared with activity with both wild and selected mutant
the activities on corncob xylan medium, both strain (Table 3).
supplemented with Tween 80. However, with
de-starched wheat bran, there’s an increase in CONCLUSIONS
enzymatic activity of 8-10%, in comparison
with the results obtained with wheat bran Acetyl xylan esterases (AXE) are key
(Table 2). accessory enzymes necessary for the complete
According to the xylanase assay, the results hydrolysis of xylan, their action increasing the
show that the highest enzymatic activity was activity of β-1.4-endoxylanases.
detected with A. brasiliensis UV 5 on both In this study, several microbial strains were
subjected to a screening regarding their ability
718
to produce acetyl xylan esterases on both Bailey M.J., Biely P., Poutanen K., 1992. Interlaboratory
corncob medium and corncob medium testing of methods for assay of xylanase activity.
Journal of Biotechnology 23 (3): 257-270.
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brasiliensis ATCC 16404 UV 7 and Penicillium Production of acetyl xylan esterase by Trichoderma
reesei and Schizophyllum commune. Canadian journal
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SCIENTIFIC PAPERS

CERES Publishing House

1 EFFECT OF INFECTION BY ILAR-VIRUSES ON VEGETATIVE GROWTH OF

1. CHARACTERIZATION OF GRAPE AND WINE QUALITY INFLUENCED BY

1 HERITABILITY OF THE TOMATO GENOTYPES RESISTANCE TO THE

FLORICULTURE, ORNAMENTAL PLANTS

1 PRELIMINARY RESEARCHON THE INFLUENCE OF THE PLANTING TIME

1 POSSIBILITIES FOR SMALL-SCALE COMPOSTING OF HORTICULTURAL

EFFECT OF INFECTION BY ILAR-VIRUSES

Institute of Agriculture - Kyustendil, 2500 Kyustendil, Bulgaria

Corresponding author email: [email protected]

Key words: influence, PNRSV, PDV, sweet cherry, vegetative growth.

INTRODUCTION combined infections, the damage is always

virus free 2.18 1.46 163.6 PNRSV 1.44 ns 241.6 ns 42.67 ns

ʻVanʼ PNRSV 1.84 ns 1.36 ⃰ 153.8 ns

MECHANICAL PROPERTIES OF THE FRUITS

Simeon KRUMOV, Nicolai CHRISTOV

Institute of Agriculture - Kyustendil, 2500, Kyustendil, Bulgaria

Corresponding author email: [email protected]

Key words: cultivar, mechanized harvesting, sweet cherry, transportability.

INTRODUCTION authors was found that the cultivar ‘Volsko’

CONCLUSIONS The sweet cherry cultivar ‘Vasinika’, created at

Viorel ENACHE, Mihaela CROITORU, Milică DIMA

Corresponding author email: [email protected]

Keys words: dynamic, annual growths, shoots, works in green.

INTRODUCTION of leaves and the foliar surface grow rapidly,

Mean length of shoots per tree (cm)

results obtained are presented in table 1. The

varieties with different ripening time and

of the apricot varieties studied in the year 2016

Adrian CHIRA1, Lenuța CHIRA1, Elena DELIAN1, Constanța ALEXE2

Corresponding author email: [email protected]

Key words: pre-harvest, post-harvest, rotten fruits.

INTRODUCTION vegetation period, but at the same time, imply a

Variety Variant Concentration Storage period Rotten fruits

Table 3. Fruits chemical analysis

‘Generos’ 80.50 19.50 14.90 0.43 4.76 0.27

Table 4. Fruits physical analysis

‘Jonathan’ 120 0.744 3.9

‘Generos’ 135 0.780 3.7

‘Golden Delicious’ 138 0.760 3.6

SYSTEMS FOR SHAPING AND PRUNING OF PLUM TREES

University of Forestry, 10 „St. Kliment Ohridski” Blvd, Sofia, Bulgaria

Corresponding author email: [email protected]

Key words: plum, pruning, forming systems.

INTRODUCTION The purpose of the currant study is to present the

The article is published in collaboration with

ASPECTS REGARDING THE QUALITY EVOLUTION OF SOME APRICOT

Corresponding author email: [email protected]

Key words: modified atmosphere, rottenness, storage condition.

INTRODUCTION MATERIALS AND METHODS

ECONOMIC EFFECT OF SUSTAINABLE APPLE PRODUCTION

Iliyana KRISHKOVA1, Denitsa SERBEZOVA2

Corresponding author email: [email protected]

Key words: apple, rootstock, yield, quality, economic analysis.

INTRODUCTION found that the density of the trees and the

ʻFlorinaʼ 2014 2015 2016 2017 Growth, cm

STUDIES ON THE BEHAVIOR OF SOME ELITE

Mihaela (ERCULESCU) STOICA, Dorel HOZA, Lenuţa Constantina CHIRA,

University of Agronomic Sciences and Veterinary Medicine of Bucharest,