ANALELE

UNIVERSITĂŢII DIN CRAIOVA

V O L . XX I I ( L V I I I ) – 2 0 17

BIOLOGIE
HORTICULTURĂ
TEHNOLOGIA PRELUCRĂRII
PRODUSELOR AGRICOLE
INGINERIA MEDIULUI

AN N AL E S OF T H E U N I V ER SI T Y OF C R AI OV A

C R AI OV A – 2 0 1 7
E d i t ur a U N I V ER SI T AR I A
 GENERAL EDITOR: Prof.dr. Sina COSMULESCU

EDITOR: Prof.dr. Violeta NOUR

SCIENTIFIC BOARD:
● Adrian ASĂNICĂ - University of Agronomic Sciences and Veterinary Medicine Bucharest,
Romania
● Adrian BACIU - Faculty of Horticulture, University of Craiova, Romania
● Valerian BĂLAN - Agrarian State University, Republic of Moldova
● Constantin BĂDUCĂ - Faculty of Horticulture, University of Craiova, Romania
● Mihai BOTU - Faculty of Horticulture, University of Craiova, Romania
● Dorin CAMEN - Banat University of Agricultural Sciences and Veterinary Medicine
Timisoara, Romania
● Daniela Doloris CICHI - Faculty of Horticulture, University of Craiova, Romania
● Vily CIMPOIAȘU - Faculty of Horticulture, University of Craiova, Romania
● Daniela CIUPEANU CĂLUGĂRU - Faculty of Horticulture, University of Craiova, Romania
● Sina COSMULESCU - Faculty of Horticulture, University of Craiova, Romania
● Marta DISZY - Universidad de la Rioja, Spain
● Ana-Maria DODOCIOIU - Faculty of Horticulture, University of Craiova, Romania
● Lucia DRAGHIA - University of Agronomic Sciences and Veterinary Medicine Iasi, Romania
● Alin DOBREI - Banat University of Agricultural Sciences and Veterinary Medicine
Timisoara, Romania
● Gheorghe GLĂMAN - President of the Romanian Horticultural Society, Romania
● Stefano GREGO - University of Tuscia, Viterbo, Italy
● Michèle GUILLOUX – BÉNATIER - Institut Universitaire de la Vigne et du Vin „Jules
Guyot”, Université Bourgogne, France
● Dorel HOZA - University of Agronomic Sciences and Veterinary Medicine Bucharest,
Romania
● Stavros LALAS - Technological Education Institution of Thessaly, Larissa, Greece
● Viorel MITRE - University of Agronomic Sciences and Veterinary Medicine Cluj-Napoca,
Romania
● Ion MITREA - Faculty of Horticulture, University of Craiova, Romania
● Mihai NICOLESCU - Romanian Academy A.S.A.S "Gheorghe Ionescu Şişeşti”, Romania
● Violeta NOUR - Faculty of Horticulture, University of Craiova, Romania
● Alexandros PAPACHATZIS - Technological Education Institution of Thessaly, Larissa,
Greece
● Jocelyne PÉRARD - Responsable de la Chaire UNESCO „Culture et tradition de Vin”,
Université Bourgogne, France
● Aurel POPA - Faculty of Horticulture, University of Craiova, Romania
● Daniela POPA - Faculty of Horticulture, University of Craiova, Romania
● Daniel RĂDUȚOIU - Faculty of Horticulture, University of Craiova, Romania
● Radu SESTRAŞ - University of Agronomic Sciences and Veterinary Medicine Cluj-
Napoca, Romania
● Felicia TUŢULESCU - Faculty of Horticulture, University of Craiova, Romania
● Liviu Cristian MĂRĂCINEANU - Faculty of Horticulture, University of Craiova, Romania

COMPUTERISED EDITING:
Doina Roşca - Faculty of Horticulture, University of Craiova, Romania

ANNALS OF THE UNIVERSITY OF CRAIOVA – DOLJ

A.I.Cuza Street, No. 13, Code 200585 – Craiova, Romania
I SSN 1 4 5 3 – 1 2 7 5
Sections:
HORTICULTURE

FOOD PRODUCTS
PROCESSING TECHNOLOGY
 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
 Food produce process ing
UNIVERSITY OF CRAIO VA
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

RESEARCH REGARDING THE INFLUENCE OF THE VARIETY AND

CULTURE CONDITIONS ON CERTAIN FRUIT QUALITY FEATURES FOR
SEVERAL CHERRY VARIETIES

Albulescu Carmen Maria Andrada1, Hoza Dorel1*

1
University of Agronomical Sciences and Veterinary Medicine Bucharest, Faculty of Horticulture; 59 Mărăști
Bvd, 011464, Bucharest, Romania,
* correspondence author. E-mail: [email protected]

Key words: fruit caliber, average weight, soluble dry substanc

ABSTRACT
Cherries are important for the consumers for the moment of appearance, being the first
fresh fruit that appear on the market, and for their quality, with important implications in the
human health. The current case study presents several aspects related to the influence of the
culture area on fruit quality. From the study of several varieties, during a time span of two years,
it could be noticed an influence of the culture area and the climatic year on certain fruit quality
features. The recorded differences were different both in terms of fruit size, average weight and
of the capacity to accumulate soluble dry substance. The same variety had different behaviors in
different areas and during different climatic years, which leads to a correct choice regarding the
assortment to be used in order to capitalize at maximum the pedo-climatic potential of the culture
area.

INTRODUCTION
Cherries are important for the consumers for the moment of appearance, being
the first fresh fruit that appear on the market, and for their quality, with important
implications in the human health (Hoza 2015). The obtained fruit production depends
on the used rootstock; generally, small rootstocks lead to a larger production (Edin et
al., 1996; Hrotko et al., 2009; Cantin et al., 2010). Fruit quality is influenced by a series
of factors among which the following can be named: rootstock used (Jimenez et al.,
2004; Cantin at al., 2010), degree of orchard intensity (Lang et al., 1997; Balmer 1998),
crown shape used in the plantation (Budan and Gradinariu 2000) and the culture area
that ensured conditions more or less proper for cherry fructification (Asănică et al.,
2011). Recently, the fruit size has been on the focus and in order to obtain high caliber
fruit standardization works are performed related to the fruit load, using chemical
substances that determine a poorer bud differentiation (Lenahan et al., 2006) or
applying chemical or physical methods to ensure the flowers would be sparse.

MATERIAL AND METHODS

The research was conducted between 2015 and 2017 on fruit from several
traditional locations for cherry culture: UASVM Bucharest, Istrița Buzău farm, SCDP
Iași, Moara Domnească farm and two other private farms from Călărași and Vrancea
counties. The assortment was not the same; during the first year 17 varieties were
analyzed while during the second year 27 varieties were used. Measurements were
5
made related to fruit size, average weight and content of soluble dry substance. In
order to measure fruit size (height, large diameter, small diameter) the calipers was
used and for measuring the average weight the precision scale with two decimals was
used. Soluble dry substance was measured using the HI 96801 refractometer.

RESULTS AND DISCUSSIONS

During the first year, the behavior of the trees was very different from one area
to another and, in general, the fruit were smaller than the ones from the second year.
Average fruit size did not vary for the same area; fruit caliber, expressed through the
large diameter, had values below the varieties‟ potential, the values recorded being
similar for the majority of varieties, with several exceptions where the values were
larger: Biggareau moreau, Ferm Red and Giant that exceeded 24 mm and Lapins with
under 20 mm (table 1).
Table 1
Influence of the variety and culture area on fruit size (2015)
Location Variety Height Large Small Average
(mm) diameter diameter wieght
(mm) (mm) (g)
Moara D. Stella 20.62 20.44 17.94 5.7
Biggareau
20.94 24.00 19.96 7.2
moreau
SCDP Iaşi Iva 20.62 22.12 18.66 5.6
HC 883515 20.32 22.28 19.06 6.3
Cociu 19.6 21.9 18.32 5.1
Ludovic 20.74 22.56 19.34 5.8
Average 20.47 22.22 18.88 5.70
UASVM Giant 20.74 22.56 19.34 6.4
Bucharest New Star 20.86 22.72 20.7 6.0
Katalin 20.46 22.98 20.38 6.8
Lapins 19.38 19.66 18.24 5.0
Kordia 22.14 22.14 18.74 5.8
Van 19.34 21.1 18.5 5.3
Average 20.49 21.86 19.32 5.88
Istriţa Ferm Red 20.94 24.38 19.2 7.2
Buzău New Star 17.94 20.78 18.18 5.0
Giant 24.58 26.5 22.32 10.1
Kordia 20.22 20.54 17.48 4.7
Early Red 18.32 22.9 19 5.4
Average 20.40 23.02 19.24 6.48
General average 20.45 22.37 19.14 6.19

Average fruit weight did not follow an hierarchy according to the biological
potential, but was influenced by the location, with the maximum value obtained for the
variety Giant at Istrița, Buzău County, of more than 10 g. and with the minimum value
obtained for the variety Kordia, also at Istrița, of only 4.7 g.
Fruit caliber, as well as weight, is extremely important for capitalization; fruit
with diameter larger than 30 mm are capitalized at very good prices. This year, there
were no such fruit obtained.
The influence of the culture location was highlighted by the comparison of
three varieties: Kordia, New Star and Giant, cultivated in two locations (fig. 1). It can be
6
noticed that for the fruit obtained within the plantation from UASVM Bucharest the
values recorded for fruit size are similar among the three varieties, but for the Istrița
plantation the values were very different, the behavior of the trees being strongly
influenced by the area.

Fig. 1 Influence of the culture area on the fruit size

During the second year of experiment, several varieties from 5 locations were
tested and. as first impression, it resulted that the climatic year was better, the fruit
being larger in all centers of origin (table 2).
Fruit height was over 24 mm for 5 of the tested varieties, while the large
diameter exceeded 27 mm for some varieties, without having any production
standardization works applied in order to determine this caliber. Among the varieties
with potential to generate large fruit one can mention Early Red, Summit, Giant and
Ferm. Fruit size was influenced by the culture area, without any correlation between
them, which means that when establishing the assortment one has to choose among
the varieties that best respond to the local conditions. Average weight was good, for
many varieties it exceeded 9 g, or even 10 g for three varieties (Grace Star, Summit
and Early Red), while the variety Giant produced at Istrița exceeded 12 g, being the
highest value recorded during the experiment.
The climatic year influenced the capacity of the varieties to accumulate soluble
dry substance. Comparing the values recorded for several varieties during the two
years, it could be noticed that there is no unitary influence for all varieties, as they react
differently to the climatic conditions of the year. Generally, the values recorded for the
first year of culture, when the fruit were smaller, were higher; however, at Istrița, two
varieties produced larger fruit during the second year, by 19.6% for the variety Ferm
Red and 5.3% for Kordia (table 3).
The influence of the area on fruit size can be better shown through the
comparison between existing varieties in different locations. Thus. for the varieties
Giant and Ferovia, the best area for culture proved to be Istrița, Buzău County, for the
variety Summit Vrancea area, while for the variety Early Red the best area is
Bucharest (UASVM) (fig. 2).

7
 Table 2
Influence of the variety and culture area on the fruit size (2017)
Variety Height Large Small Average
Location (mm) diameter diameter weight
(mm) (mm) (g)
Moara Giant
22.27 25.63 20.07 7.51
D.
Călăraşi Grace Star 23.90 26.81 26.54 10.10
Early Red 24.69 27.28 21.98 9.53
Media 24.30 27.04 24.26 9.82
Vrancea Ferovia 24.31 24.89 20.61 8.19
Summit 24.28 27.53 21.15 10.17
Biggareau
21.66 24.51 19.85 9.53
Burlat
Average 23.42 25.64 20.54 9.30
Katalin 23.24 22.98 19.81 6.95
Rubin 21.06 23.95 19.80 6.92
Lapins 21.56 22.36 19.52 7.34
Uster 21.66 21.74 18.43 6.51
UASVM Van 20.93 23.27 18.65 6.38
Bucha- Ferm Red 22.79 24.06 19.32 9.49
rest Early Red 22.51 25.13 20.89 10.70
Summit 18.53 18.23 16.08 5.32
Giant 20.66 24.11 20.23 7.38
Kordia 24.06 23.71 19.24 7.45
Skeena 21.51 23.27 19.69 6.75
Ferovia 21.20 21.16 17.92 6.28
Average 21.64 22.83 19.13 7.29
Kordia 23.20 23.58 19.79 7.81
Istriţa Van 21.62 25.44 20.39 7.67
Buzău Lapins 20.85 23.02 19.29 6.17
Ferovia 22.90 25.62 20.70 8.41
Giant 24.91 28.91 23.00 12.34
Summit 23.20 27.36 22.50 9.74
Ferm Red 23.96 27.54 21.21 9.00
Early Red 22.36 26.86 22.10 7.54
New Star 22.75 25.59 20.81 9.21
Average 22.86 25.99 21.09 8.65
Experiment average 22.90 25.43 21.02 8.51

The largest difference between the values during the two years were recorded
for the variety Giant, of 30.27%.

8
 Fig. 2 Influence of the culture area on fruit weight (g)

Table 3
Influence of the climatic year on the accumulation of soluble dry substance (%)
Location Variety 2015 2017 Difference
absolute relative
UASVM Katalin 15.9 12.84 3.06 19.25
Lapins 14.3 12.48 1.82 12.73
Van 17.1 14.82 2.28 13.33
Kordia 15.5 12.82 2.68 17.29
Average 15.7 13.24 2.46 15.67
Istrita Giant 18.3 12.76 5.54 30.27
Ferm Red 15.1 18.06 -2.96 -19.60
Early Red 15.0 13.44 1.56 10.40
New Star 16.6 15.77 0.83 5.00
Kordia 14.7 15.48 -0.78 -5.31
Average 15.94 15.10 0.838 5.26

CONCLUSIONS
From the present study, the following conclusions can be drawn: the varieties
have reacted differently from one year to another and from one location to another,
depending on their biological features; the climatic year influenced the fruit size and
content of soluble dry substance; establishing the assortment has to be done
according to the manner in which the variety responds to the climatic conditions of a
certain area, in order to obtain high quality and superior capitalization possibilities.

REFERENCES
Asănică A., Ion L., Păun C., Popa A., 2011 – Growth and fruiting of some sweet
cherry cultivars on SL64 rootstock in the South/Eastern part of Romania. ISHS
Symposium, Pitești, September 5-7.
9
 Balber M., 1998 – Preliminary results on planting densities and rain covering for
sweet cherry on dwarfing rootstock. Acta Horticulture 468:433-439.
Budan S., Grădinariu G., 2000 – Cireșul. Editura Ion Ionescu de la Brad.
Cantin C.M., Pinochet J., Gogorcena Y., Angeles M., 2010 - Growth yield and fruit
quality oj Van and Stark Hardy Giant sweet cherry cultivars as influenced by grafting
on different rootstocks. Sci.Hort., vol.123, 3:329-335.
Edin M., Garcian A., Lichou J., Jourdain J.M., 1996 Influence of dwarfing cherry
rootstocks on fruit production. Acta horticulture 41:239-243.
Hoza D. 2015 - Pomologie. Editura Ceres București.
Hrotko K., Magyar L., Gyeviki M., 2009 - Effect of rootstock on vigor and
productivity in high density cherry orchards. Acta Horticulture, 825:245-250.
Jimenez S., Garin A., Albas E.S., Bteran J.A., Gogorcena Y., Moreno M., 2004 –
Effect of several rootstocks on fruit quality of sunburst sweet cherry. Acta Horticulture
658:353-358.
Lang G.A., Ophardt D.R., Guimond C.M., 1997 - Early performance of high
density sweet cherry orchard training systems on five rootstocks. Third International
Cherry Symposium. Norwey-Danmark, Abstract. p. 60.
Lenahan O., Whiting M.D., Elfving D.C., 2006 - Gibberellic acid inhibits floral bud
induction and improves Bing sweet cherry fruit quality. Hort Science 41:654-659.

10
 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
 Food produce process ing
UNIVERSITY OF CRAIO VA
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

OPTIMIZATION OF ECOLOGICAL FACTORS AND THE INFLUENCE OF

LIGHT ON THE RATIONAL USE OF RESERVE CARBOHYDRATES IN
PERIOD DURING FORCING CUTTINGS

Bădulescu Adriana1, Ficiu Lidia2

1
National Research & Development Insitute for Biotechnology in Horticulture Stefanesti,
bă[email protected]
2
Institute of Research & Development for Viticulture and Vinification Valea Călugărească,
[email protected]

Key words: grafted cuttings, ecological factors, carbohydrates

ABSTRACT
In most countries of the world, where viticulture has millenary traditions but not only,
diseases caused by phytoplasms, viruses, viruses, bacteria and pathogenic fungi cause irreparable
damage to vineyards. The production of viticulture propagating material of higher biological categories
represents the peak area of knowledge regarding the vegetative multiplication of the vine. This
involves addressing new techniques of genetic selection (in the framework of a technically acceptable
protocol for officially accepted clonal selection), phytosanitary status verification by complementary
methods (technical protocol for phytosanitary selection), the use of rapid multiplication techniques
(especially in vitro biotechnologies) and the conservation of the vine propagation material (biological
maintenance) in the greenhouse, the classification of the activities in the certification schemes and the
unanimously accepted classification schemes.

INTRODUCTION
The new varieties and valuable clones as well as the rootstock cuttings used in
the experiments were derived from the nucleus of the isolate, which is the national
collection of viticulture germplasm made of deviated material by heat therapy and in
vitro culture (Tita and al., 2004). The biological category of the material used for
grafting was the initial propagation material. The choice of SO4-4 rootstock was
primarily due to its superior behavior on a variety of soil types and, secondly, to the
great vigor of growth it imparts to grafting varieties (Tiţa I. and al., 2001; Corbean.D.
and al., 2009). However, it is a difficult rootstock in the vine school in the sense that it
roots harder, and the triggering of calogenesis at the grafting point is delayed by 1-2
days compared to other rootstocks (Tita I., 2004). The technological improvement
aimed at experimentation was the reduction of carbonate consumption from the
biological reserve of cuttings grafted during the forcing period and after the greening of
the forced cuttings (Hamdan A-J.S. si colab., 2010).

MATERIALS AND METHODS

The preparation of the grafting material was done using the classic technology.
The grafting was performed mechanically, with DPA 5, which cuts the inverted U-
shaped cuttings, and the stratification was made in resinous sawdust, in forcing boxes
having the frame and the metal bottom, and the walls of plastic foil.

11
 The special complexity of the issues pursued necessarily implied a series of
experiments, on technological phases, all with a view to improving graft growth and
stimulating early organogenesis in order to reduce the consumption of carbohydrates
in the rootstock cuttings reserve.
The study of the quality of the grapevine material grafted during the most
critical period for symbionts was carried out in the following experimental variants:
V1 - witness, forcing into unlighted rooms
V2 - forcing into artificially lighted rooms
V3 - forwarding in non-illuminated rooms and forcing in natural light

RESULTS AND DISCUSSIONS

The experiments during the grafted period consisted of:
- determination of the physiological state of grafted cuttings (total water,
soluble carbohydrates and starch);
- protection against dehydration, oxidation of sections and pathogenic infection
of the grafting point.
These determinations were followed by complete rehydration of graft and
rootstock cuttings, initial amounts of soluble carbohydrates in carbohydrates and
starch, which were the starting points in the assessment of consumption during the
period of forcing grafted cuttings.
The experiments during the forcing of the grafted cuttings followed:
- optimal control and management of environmental factors;
- reduction of electric and thermal energy consumption;
- early and concurrent activation of bumper partners' bindings;
- stimulation of early coughing and early vascularization of the grafting point;
- stimulation of chlorophyll synthesis in young shoots of graft shoots;
- the influence of natural light on reducing the consumption of carbonated
carbohydrates and on the endogenous production of auxins involved in both the
vascularization of the grafting point callus and the early rhizogenesis decalcification.
The evolution of carbon hydrate consumption was carried out by repeated
determinations at the beginning of the vegetation phases specific to forcing grafted
cuttings (debulking, beginning of callus formation, beginning of shoots, raising of
shoots, development of callus and beginning of vascularization of grafting area)
(Bartoloni G., and al., 1996).
For the correct observation of the phenophases of growth and growth of the
shoots in the buds, the top of the grafted cuttings (the grafting point and the grafts) was
not covered with sawdust as can be seen in figure 1.

Fig. 1. Forcing cut grafted cuttings into metal casting forcing rooms and hot glass
tables in greenhouse
12
 In rooms with or without artificial illumination, the baskets are placed on the
rasters in a single row. Natural light penetrates only through the windows and only
affects the baskets in their immediate vicinity. For the rest of the baskets artificial
lighting is needed.
Early chlorophyll assimilation is more important for the subsequent
development of vines than root nutrition during the same period. The intensity of
photosynthesis is closely correlated with the intensity of the forcing spaces and the
temperature level. The highest values of photosynthesis occur at 30 ° C and 12,000
lbs.
Determinations on the amounts of carbonated carbohydrates at 10 days, 16
days, 32 days after grafting show that this technology does not consume more than
57% of the initial reserve so that a large enough amount remains to repair the organs
removed at the time classification of forced cutters (Tiţa I. and al., 2001, 2002, 2003).
In the 7 days of exposure to natural light, the assimilating pigments were
synthesized as follows:
- chlorophyll A - from 0.867 to 1.475 mg / g of dry matter representing an
increase of 58.78%;
- chlorophyll B - from 0,153 to 0,365 mg / g of dry matter, representing an
increase of 41,92%;
- carotenoid pigments - from 0.383 to 1.020 mg / g of dry substance
representing an increase of 37.55%, as shown in Figure 2

1,6
1,4
1,2
1
mg/gs.v.

0,8
0,6
0,4
21 zile
0,2
18 zile
0 14 zile
Clorofila a Clorofila b
pigm.carot

1,6
y = 0,081x 2 - 0,02x + 0,806
1,4
R2 = 1
1,2
1
0,8
y = -0,0505x 2 + 0,5205x - 0,087
0,6 R2 = 1

0,4
0,2 y = -0,041x 2 + 0,27x - 0,076
R2 = 1
0
14 zile 18 zile 21 zile

Clorofila a Clorofila b pigm.carot

Poly. (Clorofila a) Poly. (pigm.carot ) Poly. (Clorofila b)

Fig. 2. Accumulation of assimilable pigments in light shoots

13
 Analyzing the consumption of carbohydrates, from the removal of the cuttings
from the storehouse to the end of the forcing into light, there are periods when the total
carbohydrate consumptions are minimal and periods of intense respiratory and
organogenic processes resulting in maximum carbohydrate consumption cutlery
reserve.
Table 1
Content in carbohydrates at the end of forcing by the three foil techniques

Force Content in Total H of C Available Number of

technology Carbohydrates at consumption carbonates in good
the End of Force (%) cuttings (%) cuttings to
Total Starch plant in the
glucosides field
(%)
Classic 7,2 0,8 73,6 26,4 72
With artificial
lighting 7,6 1,2 52,8 47,2 76
With natural
light on hot 8,5 2,1 47,8 52,2 83
meals

Thus, during the pre-harvesting period (8-10 days after grafting), the total
carbohydrate drops from 11.7 g to 9.8 g, resulting in a consumption of about 2 g / 100
g dry material, and during the pre-emergence period in natural light (leaves and green
shoots), the consumption is only 0.1 g within 4 days.
In these two periods the consumption is minimal, in the first part only the
processes of calogenesis and deforestation are carried out, and in the second part, the
leaves being intensely chlorophylised, the photosynthesis process reduces the
consumption of the symbionts reserve.
The consumption of carbohydrates is influenced by exposure to light.
Thus, from the grafting to the greening of the shoots (14 days) the
consumption of carbonated hydrates is 5.1 g, and after the greening of the leaves and
until the planting in the vine school (15 days) the consumption is only 1.1 g although
the energy needs are maximum in part II of the force.
Percentage, at the end of forcing cuttings, by driving environmental and
technological factors, significantly reduces the consumption of carbohydrates: 56.4% of
carbohydrates are consumed in the first part of the forcing and only 8.6% in part II;
thus prior to planting in the vineyard school in the rootstock cuttings remain 52.2% of
the initial carbonate content, which represents a great advantage for the restoration of
the vines after planting in the school and for the subsequent development of the
grafted vines (Figure 3)

CONCLUSIONS
By making a comparison between the cutter force technologies in the three
experimental variants, the classic V1 (without illumination), the improved V2 and the V3
experienced at INCDBH Ştefăneşti (8-10 days darkness and forced natural light in
greenhouses, on tables warm) we can see the following:
► the highest total carbohydrate content (8.5 g / 100 g of dry material) was
recorded in cuttings forced into natural light by the technique used at INCDBH
Ştefăneşti;

14
 ►the minimum carbonate consumption (47.8% of the initial content) was
recorded in the same experimental variant, which provides the highest content in
carbohydrates (52.2%), available in rootstocks for organ recovery (roots and shoots)
destroyed during planting in the vineyard school

14
y = -0,1095x 5 + 1,83x 4 - 11,176x 3 +
12 30,45x 2 - 37,615x + 28,76
R2 = 1

10 y = -0,0404x 5 + 0,6417x 4 - 3,6162x 3

+ 8,6883x 2 - 9,5033x + 10,99
g/100g s.u.

8 R2 = 1

4
y = -0,0691x 5 + 1,1883x 4 - 7,5596x 3 +
2 21,762x 2 - 28,111x + 17,77
R2 = 1
0
Depozit La altoire La 10 zile La 14 zile La 16 zile La 30 zile

Gluc. Sol. Amidon Gluc. Total

Poly. (Gluc. Total) Poly. (Amidon ) Poly. (Gluc. Sol.)

Fig. 3. Evolution of Carbohydrate Consumption during Forcing on Merlot 8 VI / SO4-4.

ACKNOWLEDGMENT
The financing was carried out with the support of the ADER 2020 Sectorial
Program, MADR, project 3.3.9: "Maintenance of the vine propagation material - the
initial biological propagation material, Basis and Certificate".

REFERENCES
Bartoloni G., Pastelli P., Teponi M.R., Di Monte G., 1996, Rooting and
carbohydrate availability in Vitis 140 Ruggeri stem cutting. Vitis, Allemagne, no.1,
pag.11-14.
Corbean D., N. Pop, A. Babeş, A. Comşa, 2009 - Research on new methods of
forcing management for production of grafted vines at S.C. Richter Tehnologii Viticole
S.R.L. Jidvei, Bulletin U.S.A.M.V. Cluj-Napoca.
Hamdan A-J.S., R. Basheer-Salimia, 2010 - Preliminary Compatibility between
Some Table- Grapevine Scion and Phylloxera-Resistant Rootstock Cultivars, Jordan
Journal of Agricultural ciences, Vol. 6 (1), p. 1-10.
Tiţa, I., 2001. Dezvoltarea sectorului de producere a materialului săditor viticol.
Eficientizarea tehnologiei aplicată la SCPVV Ştefăneşti Argeş. Simp. Şt. Naţional
”Perspective ale restructurării şi relansării agriculturii româneşti” UŞMV Bucureşti,
pag.71-77.
Tiţa I. 2002 – Contribuţii la îmbunătăţirea tehnologiei de multiplicare rapidă a
materialului viticol din categorii biologice superioare în condiţii controlate. Teză de
doctorat. Universitatea din Craiova, p. 1-183.
Tiţa I, 2003- Staţie pilot pentru producerea materialului săditor viticol din categorii
biologice superioare, Editura Tipnaste Piteşti

15
 Tiţa I., Dumitriu. I.C., Elena Buciumeanu, Emilia Vişoiu, Smaranda Gh., Adriana
Costescu, Cătălina Guţă, 2004 – Producerea materialului de înmulţire viticol din
categorii biologice superioare”, Programul AGRAL-Cercetarea pe filiera agroalimetară
din România în contextul european, Bucureşti, ISBN 973-632-123-1, p. 209-214.
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superioare, Editura Tipnaste, Piteşti.

16
 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
 Food produce process ing
UNIVERSITY OF CRAIO VA
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

FAVORABILITY OF CLIMATIC FACTORS FROM SANDY AREAS

(OLTENIA, ROMANIA) REGARDING THE WALNUT CULTURE

Birsanu Ionescu Mariana1,*, Cosmulescu Sina Niculina2

1*
University of Craiova, Horticulture Faculty, Doctoral School of Plant and Animal Resources Engineering,
Romania; e-mail: [email protected]
2
University of Craiova, Horticulture Faculty, Department of Horticulture & Food Science, A.I.Cuza Street, 13,
Craiova, Romania; e-mail: [email protected]

Key words: walnut, climatic factors, sandy areas, favorability

SUMMARY
Walnut grows in a varied areal, but there are few areas where it yields high and constant
production, which demostrates that, besides its hereditary attributes, the ecological factors play an
important role on quantity and quality of walnut production. This study summarizes the favorability of
climatic factors on walnut culture; factors existing in sandy area of the left bank of Jiu River. In this
respect, there were used data registered during the last 55 years (1961-2016) and statistically
processed. The area that represents the aim of this study has the following attributes: the average
annual temperature of the analyzed period was 11.51℃, the average temperature during the hottest
month was 27.40℃ (registered in July 2012), the average temperature during the coldest month was -
7.58℃ (registered in January 1985), the average temperature during the coldest month (January) was
-1.42℃, while the hottest one (July) of the analyzed period was 23.38℃; the amount of annual rainfall
was 514.47 mm and the average value of aridity index was 54.99 during the same period. The sandy
area on the left bank of Jiu River, situated in the South-West of Oltenia Region, in Romania,
corresponds mostly to ecological requirements of walnut, which determined a high diversity regarding
this tree species in the area. During some passed years, climatic factors have not favoured the growth
and fructification of the tree.

INTRODUCTION
Despite the fact that walnut grows in a varied areal, there are few areas where
it yields high volume and constant production. This fact demostrates that, besides its
hereditary attributes, ecological factors play an important role on quantity and quality of
walnut production. For the fruit trees to be successful on sandy soils, climatic factors
and soil have the highest influence. Environmental factors have a direct influence on
plants, that is their growth and development depends on climatic characteristics
(Cosmulescu & Gruia 2016). Plants need a specific index of warmth during their entire
period of growth, in order to develop their vital processes: breathing, sweating,
photosynthesis, water absorption. Temperature exerts conditions on evolution of
assimilation, breathing and sweating processes, the going through different phases of
growth and fruit setting, the latent life of trees during winter rest etc. Temperature is
also very important during flowering period (Cosmulescu et al. 2010a,b). For the
success of fruit tree farming in Southern area of Oltenia region, rainfall is also
extremely important as far as the amount is concerned, as well as the period of the
year when it occurs. Prentovic et al. (2014) showed that walnut production depends
17
mostly on weather conditions, especially during fertilization and fruit growth, but it also
depends on quantity of pollen dispersed in the air. A study by Lee & Sumner (2016)
regarding the impact of climatic changes on walnut culture showed that it is one of the
most vulnerable and the projections indicated that some species‟ production can be
affected during the years with unusual warm winters. Luedeling & Gassner (2012)
show that high temperature influenced the blooming of walnut and the appearance of
leaves. Autumn frosts could affect annual growth, consequently with the loss of wood
quality and causing even the death of the tree in the end (Guàrdia et al. 2013). There is
a considerable uncertainty as far as the magnitude of potential impact of climate
change on walnuts is concerned. Walnut could be negatively affected by climate
changes due to its disadvantageous sensitivity to drought, and frost damage
manifested in the areas where the tree grows today, taking into account the predicted
temperature increase and extreme weather phenomena (Gauthier & Jacobs 2011).
Xiao et al. (2013) show that walnut has great requirements regarding temperature and
humidity indices. The predominant climatic factors which affected the growth and
development of walnut in the Sichuan Province were: average annual temperature,
cumulated temperature of ≥10℃, low annual temperature, the annual sunlight, annual
rainfall, average temperature in January and July. Han et al. (2012) conducted a
comprehensive analysis of climate and soil factors, which combines climatic conditions
and biological characteristics of walnut. The appropriate ecological indices for walnut
plantations were summarized as follows: 400~2800 m altitude, 5.5~8.5 value of soil
pH, ≥60 cm soil thickness, ≥10℃ optimal temperature, 3000~6500℃ sum of annual
temperatures, 9~18℃ annual temperature, ≥ -15℃ extreme annual temperature,
400~1200 mm annual quantity of precipitation, 300 hours/year sunshine duration, ≥-
1℃ average temperature in January, ≤28℃ average temperature in July.
This study aims to analyze the favorability of sands on the left bank of Jiu River (in
Oltenia, Romania) for walnut culture.

MATERIAL AND METHOD

Material. The weather data recorded by the Romanian National Meteorological
Institute for Meteorological Station in Bechet were used for the analysis of favorability
of climate factors regarding walnut culture in the sandy areas on the left of Jiu river.
Data recorded and processed during 55 years, 1961-2016, were used. Two time
periods were used for analysis and comparison: 1961-2013 and 2007-2016.
Method. Climate data from the Meteorological Station in Bechet were used to
calculate ecoclimatic indices for light, temperature and rainfall. Favorability of climatic
resources in the sandy area on the left side of Jiu River for walnut culture was
evaluated. In order to estimate the evolution trend of climatic parameters the
regression analysis was used, the quantification of causal relations being processed by
linear equations. To analyze the space-time variability of meteorological parameters,
the standard deviation was used and the correlation analysis was used to measure the
intensity of the link between the variables.

RESULTS AND DISCUSSIONS

Generally, ecometric indices are numerical aggregates that provide information
on characteristics of a bioclimatic phenomenon at a given location (Linster 2003) and
they are methods of analysis used to highlight the relationship between climatic
favorability or restrictiveness of a territory and its biotic envelope.
Temperature factor analysis. The average annual temperatures in Bechet
area between 1961 and 2013 varied between 10.26°C (1969) and 13.07°C (2007) with
fairly large differences from one year to the next. According to previous studies,
18
walnuts grow and develop in climatic zones where average annual temperatures are
between 9 and 10°C. Xiao et al. (2013) consider that temperature, including the
average temperature in January, the average temperature in July, the average annual
temperature and the sum of temperatures ≥10°C, had a major impact on walnut
distribution in Yunnan Province (China). Thus, temperature is the primary dominant
factor. Related to the average annual temperature, Bechet area is suitable for walnut
growth (Ionescu Birsanu & Cosmulescu 2016).
According to climatic data of the last 10 years (2007-2016), there is a tendency
to increase the average annual temperature, following the model of a straight line
whose equation is y = 0.459 x +9.403 (Figure 2). Between 1961 and 2013, the
temperature increased steadily, the trend being linear and the confidence interval
±0.4°C (Figure 1). The variation limits for the average annual temperature of the
reference period (1961-2013) were between 10.26°C (1969) and 13.07°C (2007).

Figure 1. Evolution of air temperature Figure 2. Evolution of air temperature

(annual averages) over the period 1961- (annual averages) for the period 2007-
2013 2016

Comparing the last 10 years (2007-2016) with the period 1961-2013, one can
notice that the lower limit for the average annual temperature increased from 10.26°C
(1969) to 11.40°C (2010) and the upper limit was maintained (13.07°C). In comparing
the average annual temperature for the period 2007-2016 and the average
temperature between 1961 and 2013, the differences vary between 0.19°C (2010-
2014) and 1.6°C (2007), the annual averages of the period 2007-2016 exceeding the
average of the reference range. 2011 represents an exception of these measurements.
Comparing the average monthly temperatures of the period 1961-2013 with the 2007-
2016 period, there is an increase of 0.08°C (in May), 1.36°C (in August) and 2.25°C (in
February), while for the average annual growth the rate is 0.73°C (Table 1). The results
obtained at the temperature factor analysis in the sandy area on the left side of Jiu
River, the South-West Oltenia Region reveal the tendency of temperature rise over the
last years. This growth trend is also supported by Ionita et al. (2015) for the analysis of
summer temperatures in Romania, Della Marta et al. (2007) for the summer waves in
Western Europe, Cosmulescu et al. (2010, 2015) and by Busuioc et al. (2015).
Globally, Alexander et al. (2006) report significant changes in the extreme
temperatures associated with the heating trend, especially those related to the daily
minimum temperatures during the period 1951-2003 and a generalized and significant
increase in the amount of rainfall. Overall findings of Alexander et al. (2006) are not
representative for Europe if the entire 20th century is taken into account, reporting a
small difference between the average daily and daily temperature trends (Moberg et al.
2006).

19
 Table 1
Monthly and annual average temperature during 1961-2013 and 2007-2016
Average
o
Average monthly temperatures ( C)
o
Period ( C)

I II III IV V VI VII VIII IX X XI XII (oC)

1961-2013 1.46 0.7 5.94 12.28 17.81 21.45 23.36 22.56 17.8 11.5 5.5 0.3 11.47

2007-2016 -0.59 2.95 7.12 13.01 17.89 22.07 24.44 23.92 18.69 11.26 6.41 1.44 12.2

Differences -2.05 2.25 1.18 0.73 0.08 0.62 1.08 1.36 0.89 -0.24 0.91 1.14 0.73

Klein Tank & Konnen (2003) reported similar results between 1946 and 1999,
respectively a small difference between the average daily and minimum daily
temperature trends. In the Mediterranean Basin, extreme temperatures are in line with
global trends: the minimum temperature decreases and the maximum increases
(Efthymiadis et al. 2011). For walnut culture, Xiao et al. (2013) reported the following
optimal temperatures in Yunnan: 9.19ºC average temperature in January, with
variations between 6.23-12.16ºC, 21.42ºC July average temperature, with variations
between 19.08-23.76ºC and average annual temperatures of 16.36ºC. Solar et al.
(2008) determined how winter and spring temperature could affect the walnut (Juglans
regia L.). Phenological models showed that the average air temperature in the two
months (March and April) was important for the estimation of deforestation.
Rainfall analysis. From the analysis of data on the amount of rainfall recorded
in Bechet area during period 1961-2013 it is observed that the highest values of annual
average are registered in 2005, with an average value of 811.83 mm. The lowest
rainfall were recorded in 2000 (288.24mm), considered to be dry in terms of rainfall.
The multi-annual average (1961-2013) of rainfall in Bechet area is 516.92 mm, being
below the lower limit of walnut need (minimum 600 mm annual rainfall). During the last
10 years (2007-2016) annual average values ranged between 349 mm (2016) and 686
mm (2010). Also, the multiannual rainfall average in this area over the last 10 years
(508.73 mm) is below the lower limit of walnut need. Botu et al. (2014) investigated the
horticultural performance of 10 native varieties and eight varieties of walnuts
introduced from other countries cultivated in the Northern region of Oltenia. It was
found that the environment contributed with 24.44 and 29.1%, respectively, and the G
x E interaction with 12.22 and 14.6%. For a synthetic image of the space-time
variability of rainfall variance, we calculated the deviations of averages of annual
rainfall quantities registered during 2007-2016 compared to the annual rainfall
averages in the reference period 1961-2013. In Table 2, one can see that positive
values of deviations of annual rainfall quantities were recorded in 2007 (20.97 mm),
2009 (91.52 mm), 2010 (169.59 mm), 2014 (100.89 mm) and negative values in most
of the years: 2008 (-72.05 mm), 2011 (-139.84 mm), 2012 (76.66 mm), 2013 (818.82
mm), 2015 (69.99 mm), 2016 (167.54 mm).
Lately there have been drought problems in many agricultural areas.
According to the National Meteorological Agency (NMA) in the high risk area of drought
and desertification, the climate is warm and dry, with annual average temperatures
above 10ºC, and the average annual rainfall is between 350-550 mm, of which the
amount between 200-350 mm is during vegetation period (April to October).

20
 Table2
Difference between annual average rainfall and average rainfall from 1961 to 2013
Sum of annual average Sum of average rainfall from Difference
Year rainfall (mm) 1961 to 2013 (mm) (mm)
2007 537.55 516.58 20.97
2008 444.53 -72.05
2009 608.1 91.52
2010 686.17 169.59
2011 376.74 -139.84
2012 439.92 -76.66
2013 497.76 -18.82
2014 617.47 100.89
2015 446.59 -69.99
2016 349.04 -167.54

Based on these criteria, an analysis of Bechet area was made on the risk of
drought manifestation. Annual average temperatures exceed 10°C. For rainfall, the
sum of annual rainfall was also calculated from April to October. From this point of
view, the years 1983, 2002, 2007 and 2016 were years of high risk of drought, the sum
of rainfall being below 350 mm, and the average annual temperatures of 11.57ºC,
12.72ºC, 13.07ºC and 12.16ºC, respectively. A parameter that expresses the
correlation between climate, vegetation and the waters of a territory is considered
aridity index, established by Martonne. It expresses the relationship between
multiannual rainfall and multiannual average temperature. This index makes it possible
to determine the aridity of a region for characteristic periods (a year or a month), being
an expression of the restrictive nature that climate conditions impose on certain plant
formations (Satmari 2010). The analysis of aridity indices by Martonne and UNEP
reveals the obvious tendency of aridization in S-V Oltenia over the last three decades,
and since 1980 a semiarid climate has been established in the area, with the aim of
escalating aridisation (Pravalie et al. 2013). According to climatization of the region,
according to the provisions of Balteanu et al. (2013) Southern Oltenia is characterized
by a temperate continental climate of transition with Mediterranean influences. The
average annual temperature is 10ºC (over 11ºC in the Danube Meadow), precipitation
ranging from 500-600 mm / year, air humidity around 78-80%, specific climatic
characteristics for the sandy, southern plains, its individuality being due to aridity and
drought phenomena. The annual average evapotranspiration potential is over 700
mm/year (Sandu et al. 2008; Dragota et al. 2011). The space distribution of the values
of Martonne aridity index on the territory of Romania has been the object of several
researches (Dumitrascu 2006; Paltineanu et al. 2007a,b; Pravalie 2014). These
researches show that the lowest (extreme) values, below 20, are in eastern Dobrogea
below 25 in the Danube (Oltenia), in the East of the Romanian Plain and West
Dobrogea; also in the Plain of Oltenia and the Center of the Romanian Plain the values
are between 25-30. For plant culture, the amount of rainfall in the growing season is
important. The lack of rainfall from a certain period can not be compensated for by
surplus rainfall in another period. Between 1961 and 2016, in Bechet area there were
months of very low, insignificant rainfall: 0.14 (October 1969), 0.25 (January 1989) or
months with rainfall over the multiannual average of the month (August 2005 -
200.19mm, October 2007 - 106.99 mm, September 2014 - 111.50 mm). Correcting
with the literature and the requirements of the fruit tree species, it is found that the area
is suitable for crops with low requirements (at least 500 mm annual rainfall, peach,
apricot, almond), but also to medium-sized species (minimum 600 mm annual rainfall:
pear- tree, walnut, cherry, cherry, summer apple varieties). Average rainfall of 700 mm
or above is exceptional (year 2005-811.83), so it is necessary to irrigate the crops of
21
large species of fruit (quince, late varieties of apple, plum) with high and very high
need for water (fruit shrubs). Walnut requires areas with average annual rainfall of at
least 600 mm. The data analysis shows that in the Bechet area, only 10 of the 55
calendar years analyzed correspond to walnut requirements. Throughout the
vegetation period there are phenophases in fruit growing which require an appropriate
level of humidity. One of these is the growing period of shoots and/or the fruit, when
the requirement of the trees for water are high. The period of intense growth of shoots
and fruits is May - July. For a good development of these phenophases, we are
interested in the amount of rainfall during this period. The analysis of the average
precipitation in May - July, in the last 55 years (1961-2016) shows great variability,
from 22.24 mm (1985) to 108.63 mm (1970), and in the last 10 years from 28.19mm
(2007) to 89.19mm (2010). The average rainfall for May in the first and second interval
of the month is 57.19mm, respectively, 64.27 mm, for June 61.55mm, respectively
71.43 mm, and for July 52.85mm, respectively, 46.45mm. The amount of rainfall from
May to July differs from one year to the next, the variation limits in the first interval
(1961-2013) being as follows: between 11.2mm (2000) and 111.01mm in May (1967);
between 9.57mm (2003) and 126.66mm in June (1966); between 6.35 (1965) and
165.56mm in July (1970). In the second range (2007-2016), the variation limits are:
between 32.80 mm (2008) and 107.14 mm in May (2010); between 25.82 mm (2007)
and 117.35 mm in June (2010); between 10.11 (2007) and 102.09 mm in July (2009).
It is necessary to analyze whether rainfall was useful for growing and
developing trees. Useful rain is considered to be over 10 mm. To ensure a sufficient
wetting of soil with balanced hydrophobic characteristics, the amount of rainfall (in mm)
of each month should be three times more than the average monthly temperature
(Mihaescu, 1998). In order to highlight whether the rainfall was useful for the growth of
shoots and fruits, the ratio between the annual average rainfall and the average annual
temperature for the period 1961-2013 and May-July for Bechet area was calculated.
According to the data obtained, not all the rainfall during the analyzed months and
years were useful. Values smaller than 3, for the three analyzed months, were
obtained in 1985 and 2000, being the years with the lowest aridity index (46.20 and
32.66 respectively). For other years, the variation limits for this report are very high:
from 1.38 to 6.28 in 1961; from 3.70 to 7.37 in 1970; from 1.23 to 6.90 in 1980; from
2.92 to 7.24 in 1991; from 1.88 to 7.71 in 2004 etc. Of total values obtained after
studies, for the 55 years, from May to July, about 34% situations have a value greater
than 3, which means that only 34% of the analyzed cases have achieved sufficient
wetting for the growth and development of fruit trees. Analyzing the values obtained for
each month we can conclude the following: about 49% of the values obtained in May
are over 3, about 40% in June and about 24% in July.
It is noted that irrigation is recommended for good growth and development of
plants due to the fact that the unevenly distributed rainfall during the year does not
provide the necessary moisture for certain vegetation phenophases. In order to
mitigate the impact of climate change and to prevent land degradation, reconstruction
of the forest protection belts created between 1970-1980 is a necessity as far as they
are no longer able to protect the sandy lands (Achim et al. 2012).

CONCLUSIONS
The data from the recent studies reffering to the average temperature increase
during the last years confirmed a value of 12.20ºC (2007-2016) compared to 11.49ºC
between 1961 and 2013. Temperatures above 35°C (which cause thermal stress) were
recorded each year in July-August between 2007 and 2016, with the exception of 2010
and 2014, the absolute maximum temperature (44.08°C) being reached in July 2007.
22
Temperatures below zero degrees were recorded in: April 1970 (-2.44°C), 1974
(-2.00°C), 1993 (-1.34°C), 1997 (-2.53°C), 2000 (-1.57°C) 2001 (-1.18ºC), 2002
(-2.73ºC), 2003 (-1.71ºC), 2011 (-1.48ºC), 2012 (-2.06ºC) after starting in vegetation.
The multiannual average rainfall in the Bechet area is 514.47 mm, a value which is
below the walnut limit (minimum 600 mm annual precipitation). In conclusion, the sandy
area on the left side of the Jiu River corresponds largely with the ecological requirements of
the walnut, which made the diversity of walnuts in this area to be quite large. Nevertheless,
sometimes climatic factors are unfavorable to growth and fructification.

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24
 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
 Food produce process ing
UNIVERSITY OF CRAIO VA
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

EXTRACTS OF HERBS AND SPICES AS NATURAL ANTIOXIDANTS FOR

IMPROVING THE FUNCTIONAL VALUE OF MEAT PRODUCTS

Boruzi Andrei Iulian1, Nour Violeta2*

1
"Dunarea de Jos" University of Galati, Romania
2
University of Craiova, Romania
*Corresponding author e-mail: [email protected]

Key words: functional foods, extracts, oils, powders, herbs and spices, natural antioxidants

ABSTRACT
Lipid oxidation is one of the causes of deterioration in meat and meat preparations as it is
accompanied by the appearance of a large number of undesirable changes in flavor, texture and
nutritional value. The rate of lipid oxidation can be effectively reduced by the use of antioxidants.
Synthetic antioxidants have been widely used in the meat industry, but consumer concerns about
product safety and toxicity have prompted the food industry to look for natural sources. Natural
antioxidants such as extracts of rosemary, oregano, sage, green tea, cloves, garlic, citrus peel and
seeds, lotus, barley and Moringa oleiferia leaves can be used as alternatives to synthetic antioxidants
because of their equivalent or even higher effect on inhibition of lipid oxidation.

INTRODUCTION
Consumers nowadays attach greater importance to all aspects that can help
improve the quality of life; and diet, though not the only element that influences well-
being and health, is one of the most important. The factors that favored this evolution
include the current high impact on public opinion of media reports on the relationship
between diet and health, increasing the life expectancy of the population (this
generates high purchasing power consumers with higher health problems and are very
motivated to participate in initiatives aimed at maintaining health), an attentive attention
to disease prevention, etc.
Lipid oxidation is the major reaction that affects the flavor, color, texture and
nutritional value of food (Kanner, 1994). Various synthetic antioxidants such as butyl
hydroxytoluene (BHT), butylhydroxyanisole (BHA) and tert-butylhydroquinone have
been used to prevent oxidative damage to food. However, synthetic antioxidants are
not fully accepted by consumers because of health concerns. Therefore, some natural
ingredients, including herbs and spices, have been studied, especially in Asian
countries, as potential antioxidant products in meat and meat products (McCarthy et
al., 2001).
Herbs and spices contain many phytochemicals that are potential sources of
natural antioxidants including phenolic diterpenes, flavonoids, tannins and phenolic
acids (Dawidowicz et al., 2006). These compounds have antioxidant, anti-inflammatory
and anti-cancer activities. In food systems, they can improve the flavor, delay
degradation of foods induced by lipid oxidation, inhibit the growth of microorganisms,

25
and play roles in reducing the risk of certain diseases (Achinewhu et al., 1995, Tanabe
et al., 2002).
Among these spices, it is said that cloves have the strongest antioxidant
capacity, followed by rose petals, cinnamon, nutmeg and other spices (Al-Jalay et al.,
1987). In addition, spices have an antimicrobial capacity mainly due to phenolic
compounds. Possible mechanisms for the antimicrobial effect of phenolic compounds
include: modification of microbial cell permeability (Bajpai et al., 2008) interfering with
membrane function, including electron transport, absorption of nutrients, protein and
nucleic acid synthesis, and enzymatic activity (Bajpai et al. 2008), the interaction with
membrane proteins that cause deformation in structure and functionality (Rico-Munoz
et al., 1987) and substitution of alkyls in the phenol nucleus (Dorman & Deans, 2000)
(Zhang et al., 2010).

Rosemary extracts
The rosemary extract contains high levels of phenolic compounds that lead to
high antioxidant activity. Phenolic compounds are capable of regenerating endogenous
tocopherol in the phospholipid bipolar lipoprotein (Rice-Evans et al., 1996). Sebranek
et al. (2004) reported that extracts of rosemary added to pork sausages at 2500 ppm
were the same or even more effective than BHA / BHT in delaying TBARS values in
raw and chilled sausages during refrigeration and freezing. In addition, the addition of
rosemary extracts has improved the color and freshness of pork sausages (Sebranek
et al., 2004). Yu et al. (2002) added water-soluble extracts of rosemary in cooked
turkey products and found they were effective in slowing lipid oxidation and preventing
color loss evidenced by L-value decrease and increase in a* during refrigeration. In the
case of pork, the combination of oleoresins from rosemary with tocopherol effectively
reduced volatile hexane without inducing any effect on the production of sulfur volatiles
(Nam et al., 2006). Rosemary extracts have led to better color retention, evidenced by
decreased metmyoglobin concentration and increased oxymyoglobin values on
storage for 8 days in beef minced meat (Zhang et al., 2010).

Green tea
Catechins represent a predominant group of polyphenols present in green tea
leaves, composed of four epicatechin compounds, epicatechin gallate, epigallocatechin
and epigallocatechin gallate (Zhong et al., 2009). These tea compounds promote
health by preventing lipid oxidation and providing antibacterial, anticancer and antiviral
capabilities (Katiyar & Mukhtar, 1996; Yang et al., 2000). Tea catechins are known to
reduce peroxide formation even more effectively than alpha-tocopherol and BHA in
chicken and pig fat (Chen et al., 1998). Tea polyphenols may inhibit the formation of
mutagenic agents known to be associated with breast and colon cancer (Weisburger et
al., 2002).
Adding 300 ppm catechins from tea significantly reduced the TBARS values
for beef, duck, ostrich, pork and chicken during refrigeration for 10 days. At the same
concentration, tea catechins provided an antioxidant capacity of two to four times
higher than alpha-tocopherol, depending on the meat of different animal species (Tang
et al., 2001). Green tea extract reduced TBARS formation and the concentration of
putrescine and tyramine in dried fermented sausages. The addition of green tea,
however, had no significant effect on the pH, color, and overall sensory quality of
sausages (Bozkurt, 2006). In the case of pork sausages, green tea powder could
partially replace the nitrite, resulting in a lower TBARS value and a low volatile basic
nitrogen content compared with samples prepared with nitrite only (Choi et al., 2003).

26
 Cloves
Cloves (Eugenia caryophyllus) are known to have long-lasting antimicrobial
activity due to their active ingredient - eugenol (Cort, 1974). Cloves oil at 0.5% and 1%
inhibited the growth of L. monocytogenes in minced sheep meat. At 1%, the number of
L. monocytogenes decreased by 1-3 log cfu/g in sheep meat (Menon & Garg, 2001). In
ready-made chicken sausages, clove oil at 1% and 2% inhibited growth of L.
monocytogenes during storage at 5 °C and 15 °C (Mytle et al., 2006). Clay oil was also
effective in inhibiting other food-borne pathogens including C. jejuni, S. enteritidis,
Escherichia coli and Staphylococcus aureus (Smith-Palmer et al., 1998). Cloves were
able to prevent discoloration of raw pork during storage at room temperature and were
the most potent antioxidant in the lipid oxidation delay of all spices and herbal extracts
used, including cinnamon, oregano, pomegranate, and grape seed (Shan et al., 2009).
In another study, adding cloves oil in combination with lactic acid or vitamin C could
reduce lipid oxidation, maintain color and improve the sensory properties of buffalo
meat during store sales (Naveena et al., 2006).

Garlic
Allicin is known as the main ingredient of garlic having antimicrobial activity
against gram-positive and gram-negative bacteria. Allicin is produced enzymatically
from its algin precursor through the intermediate product of allylsulfenic acid (Ellmore &
Feldberg, 1994). Many studies have shown that garlic extract has been effective in
reducing the growth of many pathogens including S. aureus, S. albus, S. typhi, E. coli,
L. monocytogenes, A. niger, Acari parasitus, Pseudomonas aeruginosa and Proteus
morganni (Kumar & Berwal, 1998; Maidment et al., 1999).
In refrigerated chicken meat, aqueous garlic extract inhibited the growth of
microbial contaminants, including aerobic, mesophilic and feces coliforms on the
surface of poultry carcasses (Oliveira et al., 2005). The addition of 1% and 3% garlic
juice could lead to a decrease in the value of peroxide, TBARS, residual nitrite and
total number of microorganisms in sausages during cold storage compared to the
control group (Park & Kim, 2009).

Sage
Sage is commonly used in pork sausages. The main antioxidant compounds in
sage include carnosol, carnosic acid, rosmadial, rosmanol, epirosmanol and methyl
carnosate (Cuvelier et al., 1994). The addition of essential sage oil (3%) resulted in a
decrease in TBARS in both raw and cooked pork meat by 75% and 86% respectively,
whereas in raw beef and veal they fell by 57% and 62% % compared to the control
(Fasseas et al, 2008). The sage extract, alone or in combination with sodium
isoascorbate, reduced water and pH activity, reduced the number of mesophilic
bacteria and coliforms in vacuum packed turkey meat but had a better taste in cooked
sauces (Karpinska-Tymoszczyk, 2007). In high-pressure processed chicken, Salvia
protected chicken breast from lipid oxidation during refrigerated storage for 2 weeks
(Mariutti et al., 2008).

Oregano
Oregano is a traditional Mediterranean spice, and the essential oil of oregano
obtained from the steam distillation process contains more than 30 compounds.
Among the compounds, carvacrol and thymol contribute most to its antioxidant
capacity (Vekiari et al., 1993). Pig meat and beef to which 3% essential oil of oregano
was added showed lower levels of oxidation after 12 days of refrigerated storage
(Fasseas et al., 2008).
27
 Oregano oil could prolong the shelf life of fresh chicken meat by reducing the
growth of microorganisms during storage in refrigeration. However, 1% of oregano oil
could introduce a very strong unfavorable flavor to food, which would lead to low
sensory quality (Burt, 2004, Chouliara et al., 2007). The oregano essential oil (0.05%,
0.5% and 1%) could delay the development of microorganisms and decrease the final
number of alteration microorganisms under modified atmosphere conditions
(Skandamis & Nychas, 2001; Zhang et al. 2010).

Lotus and barley leaves

Lotus (Nelumbo nucifera) is widely cultivated in Asia and grows in water (Kim
& Park, 2008). It contains abundant dietary fiber, vitamins, riboflavin, potassium and
copper (Chiang & Luo, 2007). It also contains antioxidant compounds such as ascorbic
acid and phenolic compounds, carotenoids, flavonoids, phenolic acids, and
tocopherols (Hertog, 1994). The free-radical scavenging activity of phenolic
compounds from lotus leaves has been reported on by Park et. al. (2007), and the
antioxidant activity of lotus leaf extract has been described by Choe et al. (2010).
In a recent study it has been confirmed that barley leaf extract has antioxidant
activity, reducing power, free-radical scavenging activity, and superoxide dismutase-like
activity (Choe et al., 2010). It has been demonstrated via test results that in cooked
ground pork during refrigerated storage for 10 days the additions of lotus leaf powder
(LP) and barley leaf powder (BP) had positive effects on oxidative and color stability.
The ground pork made with addition of LP and BP had lower TBARS values, POVs
and CD compared to control (−) during storage. The addition of BP had an inhibitory
effect on lipid oxidation when compared to the control (−) over the 10 day period. The
addition of LP and BP has shown no significant differences in the overall acceptability.
Widely used vegetables in most Asian countries, lotus and barley leaf extracts, appear
to be useful ingredients in ground pork, providing a source of natural antioxidants that
are safe and enhance the oxidative stability of the processed meat (Choe et al., 2011).

Moringa oleiferia leaves

A good source of phenolic compounds and a very potent antioxidant activity is
found in mature M. oleiferia leaves (MOL). Cooked goat meat patties could be
protected against lipid oxidation during refrigerated storage by incorporating 0.1%
extract of MOL (100 mg⁄ 100 g meat). The MOL extract was more effective than BHT in
maintaining low TBARS number of precooked chilled goat meat patties. Frozen ready-
to-eat meat products are available in Indian cities, but freezing is quite expensive
leading to limited market. Therefore, MOL-treated meat products that can be stored at
chilled temperatures would be beneficial for the manufacturer as well as the consumer.
A potent source of phenolics and an immense nutraceutical value for the development
of functional meat products of commercial interest are the unutilized mature Moringa
leaves (Das et. al 2012).

Pomegranate
Native from Iran, pomegranate (Punica granatum) is now also cultivated in
several provinces in China. Pomegranate rind and seeds are byproducts obtained
during processing of pomegranate juice. It is reported that pomegranate rind,
pomegranate juice, and pomegranate seeds possess significant antioxidant activity
due to polyphenolic compounds (Fischer et al, 2003). The use of pomegranate juice,
pomegranate seed powder, and pomegranate rind powder has been recently
investigated as natural antioxidant in chicken and goat meat (Naveena et al, 2008;
Devatkal et al, 2010).
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 CONCLUSIONS
Meat and meat products are fundamental components of our diet, contributing
significantly to the intake of various nutrients and substances such as proteins, fats,
fatty acids, cholesterol, sodium, nitrites, etc., which have various health implications.
Meat and meat products could be protected against lipid oxidation by incorporating
extracts, oils or powders from herbs and spices in order to maintain their color and to
improve their sensory properties.

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 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
 Food produce process ing
UNIVERSITY OF CRAIO VA
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

CHARACTERIZATION OF THE ON-FARM PLUM COLLECTION FROM

PLENIŢA - DOLJ COUNTY AND THE DEVELOPMENT OF GENETIC
RESOURCES IN ROMANIA
1,2* 3,4 1 3,4 2
Botu Mihai , Pamfil Doru , Botu Ion , Hârţa Monica , Scutelnicu Anca , Giura
2 5
Simona , Popa Mihai
1*
University of Craiova, Faculty of Horticulture, Department of Horticulture and Food Science, Craiova
2
University of Craiova – SCDP Vâlcea, Rm. Vâlcea
3
University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca,
Faculty of Horticulture, Department of Horticulture and Landscape, Cluj-Napoca
4
University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca,
Life Sciences Institute, Research Centre for Agricultural Biotechnology, Cluj-Napoca
5
S.C. Secerișul S.R.L., Plenița
* Correspondence author. E-mail: [email protected]

Key words: germplasm, genetic resources, conservation, Prunus

ABSTRACT
During 2014-2017 period, regeneration of a part of national plum collection from UCv-
SCDP Vâlcea have been carried out. Through GERMPLUM project, an on-farm plum collection
consisting in 141 accessions of those regenerated has been established in Pleniţa – Dolj county
at S.C. Secerişul S.R.L. The 71 original and 70 foreign accessions from the on-farm collection
belong to 13 species of Prunus genus, 5 interspecific hybrids and 2 others. P. domestica (51.8%
of the accessions), P. insititia (22.7%), P. salicina (7.1%) and P. cerasifera (5.0%) hold the share
in the collection.
The plum accessions consist in 96 improved cultivars (34 autohtonous and 62
foreign), 11 old native cultivars, 26 biotypes from different populations and 8 wild species.The
accessions were phenotypically characterized using 31 different accepted descriptors, variability
of characters being observed.
The genetic resources present into this collection are valuable and can be used into
the future plum breeding programs for cultivars and rootstocks

INTRODUCTION
Identifying and maintaining biodiversity in fruit tree crops has become an
important necessity for assuring the survival conditions of future human generations.
Numerous plum collections hold over 5447 accessions of Prunus domestica,
802 accessions from P. salicina and over 200-300 accessions from P. cerasifera, P.
americana, etc. (Botu, 1999).
Ramming et al. (1992), Mehlenbacher et al. (1992); Botu et al. (2005); Butac at
al. (2011), Botu et al. (2012) highlights the value of plum biodiversity, how to conserve
genetic resources and their importance in breeding programs and for future
generations.
Due to the high genetic variability among the over 40 species and interspecific
hybrids (diploid, triploid, tetraploid, pentaploid and hexaploid) of plum used more or
less in culture, but also the variability existing within the same species, it is necessary
33
to save this genetic patrimony from its loss through genetic erosion and genetic
vulnerability and to value it.
World plum production amounts to more than 11,282,527 t, which corresponds
to only 1.6 kg of plums/year/capita (FAO. 2016). This consumption is considered to be
insufficient for a normal diet. Several global organizations are concerned with
preserving and making good use of plum genetic resources (European Prunus
Database, Biodiversity International, FAO, UPOV, etc.).
The value of plum genetic resources is that they are very well adapted to local
environmental conditions, quite rustic and productive and have resistance genes for
most dangerous diseases, they are productive and quite rustic.
During 1994-2001 period and through the GENRES - 61 Project of the
European Cooperation Program on Genetic Resources (ECP/GR)- Program 1467/94
on Prunus species also the Prunus genetic resources from Romania were investigated.
292 original accessions and 428 accessions introduced from other countries were
recorded from the national Prunus collections (Botu et al., 2001).
Later on, a part of the plum genetic resources from national collections were
characterized both by phenotypic and genetic descriptors through a national project
PN-II-PT-PCCA-2013-4-1399 no. 168 in titled ”Conservation of native plum germplasm
based on a experimental model of molecular characterization and collection on farm”
(GERMPLUM).

MATERIAL AND METHODS

The plum biological material consists of native genotypes from Romania
(original genetic resources) and foreign genotypes existing mainly in the national
collection, which needs to be regenerated due to the age of the trees. The plum
genetic resources in the national collection include both improved varieties, old
varieties, populations (biotypes), species and hybrids.
The biological material was grafted at UCv-SCDP Vâlcea onto selected
Myrobalan seedlings („Mirobolan galben‟rootstock). Planting was done in the autumn of
2016 and spring of 2017 on farm land belonging to S.C. Secerişul S.R.L. located in
Pleniţa, Dolj county (SW of Romania). Planting distances used were 5.0 x 4.0 m
(density of 500 trees/ha).
The phenotypic characterization of the accessions was carried out using the
internationally accepted descriptors of Biodiversity International (former IPGRI), F.A.O.
and EURISCO. From a total number of 47 descriptors used for plums, 31 descriptors
were selected in order to be used to characterize the plum accessions.
In the present paper only the passport and the general descriptors specific to
the phenotypic characterization are used, molecular characteristics will be presented
later.
The phenotypic characterization of accession will serve for setting up a
computerized data bank.

RESULTS AND DISCUSSIONS

During 2014 - 2017 period the project PN-II-PT-PCCA-2013-4-1399 no. 168 in
titled ”Conservation of native plum germplasm based on a experimental model of
molecular characterization and collection on farm” (GERMPLUM) have been carried
out. The consortium of the project was composed by University of Agricultural
Sciences and Veterinary Medicine (USAMV) from Cluj-Napoca as coordinator and
University of Craiova – SCDP Vâlcea and S.C. Secerişul S.R.L. as partners. The
project had as main objectives the identification, evaluation, multiplication and
establishment of an on-farm plum collection.
34
 During this period, 185 genotypes were identified, out of which 115 are original
(native) and 70 foreign. Following the multiplication and establishment of the 2.26 ha
on-farm collection in Pleniţa 141, accessions are found in this collection, of which 71
are original and 70 are foreign (Table 1). The accessions belong to a number of 13
species, to which are added 5 interspecific hybrids and 2 others.
Table 1

The on-farm plum collection from Pleniţa Dolj and the membership of accessions to different
species of the Prunus genus

Total no. of Accessions Out of them: Accessions

No. Species accessions introduced into identyfied but
identyfied the Pleniţa not introduced
collection into the Pleniţa
No. % No. % Autochto- Foreign collection
nous
1 P. domestica 98 53,0 73 51,8 34 39 25
2 P. insititia 43 23,2 32 22,7 27 5 11
3 P. cerasifera 13 7,0 7 5,0 6 1 6
4 P. spinosa 3 1,6 2 1,4 2 - 1
5 P. salicina 10 5,4 10 7,1 - 10 -
6 P. sibirica 1 0,6 1 0,7 - 1 -
7 P. americana 3 1,6 3 2,1 - 3 -
8 P. microcarpa 1 0,6 1 0,7 - 1 -
9 P. nigra 1 0,6 1 0,7 - 1 -
10 P. besseyi 1 0,6 1 0,7 - 1 -
11 P. maritima 1 0,6 1 0,7 - 1 -
12 P. tomentosa 2 1,1 2 1,4 - 2 -
13 P. subhirtella 1 0,5 1 0,7 - 1 -
14 Interspecific 5 2,7 4 2,8 2 2 1
hybrids
15 Others 2 1,1 2 1,4 - 2 -
Total 185 100 141 100 71 70 44

Of the 141 accessions in the collection, 73 of them (51.8%) belong to P.

domestica, 32 (22.7%) to P. insititia, 7 (5.0%) to P. cerasifera, 2 (1.4%) to P. spinosa,
10 (7.1%) to P. salicina, etc.
Through the GERMPLUM Project, efforts have been made to save a part of
the plum biodiversity, which is greatly affected by erosion and genetic vulnerability.
Also, we tried to overcome the negative effects produced in the old national plum
collection from UCv-SCDP Vâlcea due to the age of trees, as well as the loss of some
of the accessions due to land restitution to previous owners. At the time of counting of
plum accessions through the GENRES - 61 Project of the European Cooperation
Program on Genetic Resources (ECP/GR) - Program 1467/94 the UCv-SCDP Vâlcea
collection was holding 292 original (autohtonous) and 428 introduced (foreign)
accessions.
With the help of GERMPLUM project, 141 plum accessions from UCv-SCDP
Vâlcea were regenerated and introduced into the on-farm plum collection in Pleniţa.
The status of these accessions is presented in Table 2. Original or autochtonous
accessions (71 in total) contain 34 improved cultivars, 11 old cultivars and 26 biotypes
of different populations. Introduced or foreign accessions from the on-farm collection in
Pleniţa consist in 62 improved cultivars and 8 Prunus species.
All the 141 plum accessions are presented and recorded based on passport
and general descriptors so that they can be registered in the national and foreign data
banks. 31 descriptors were used to characterize each of the 141 plum accessions.

35
Along with the recording of the descriptors, 141 phenotypic characterization sheets
were written, out of which the limits of variation of the most important accessions
characteristics are presented in Table 3.
The phenotypic variability will then be correlated with the molecular data
issued as result of SSR analysis carried out at USAMV Cluj - Napoca.
A very high genetic variability was observed among the plum species
belonging to Prunus genus, but the variability is also present within the same species.
This variability will later serve in the breeding program for the obtaining of valuable
cultivars and rootstocks based on the genes available in this germplasm fund. Table 3
shows the variability of 10 characters (growth vigor, fruit size, tolerance to Plum-pox
virus, etc.) present in these accessions.
Tree growth vigor varies between 1.2 m in tree height (P. tomentosa) and 7.0
m (Marianna GF 8-1). The pollination capacity is differentiated from the level of male-
sterility (Tuleu gras) to that of self fertility (Gogoşele negre, Oteşani 8, etc.), and the
flowering time occurs between early March (P. sibirica) and early May (Vânăt
românesc cl. 4).
The size of the fruit is a very variable character, fruits vary from 4.0 to 6.0 g
(Spinigra) up to 120 g or more (Ozark premier).
Fruit yield is very variable in the case of the given plum accessions. The yields
start at a level of 0.2 kg/tree at Spinigra (interspecific hybrid) and reaching up to 60.0
kg/tree at Centenar and Stanley cultivars.Some accessions are used as generative or
seedling rootstocks (C 163, Oteşani 8, Voineşti B), others as vegetative or clonal
rootstocks (Oteşani 11, Miroval).
The range of resistance / tolerance to Plum-pox virus (PPV) attack is very
wide. Dobrovica and other accessions proved very susceptible to this attack, but
Andreea, Romandreea and Scolduş are tolerant and even exhibiting field resistance to
PPV.
Because temperatures in some areas of Romania are sometimes going below
-29°C; -31°C, some cultivars from P. salicina (like Methley) proved less adapted to the
climate.
The on-farm plum collection from Pleniţa is in the 2nd year (2nd leaf) after
planting. In the next coming years there is need for financial and scientific support.
Also, the activities of identification and promotion of new plum accessions has
to be continuated in order to enlarge the germplasm collections and to use the genetic
diversity for breeding new cultivars and rootstocks.
The collection serves for research activities and also for didactical purposes for
undergraduate and graduate students but also for research and teaching staff.

CONCLUSIONS
During 2014-2017 period, based on the GERMPLUM Project, a on-farm plum
collection made up of 141 accessions was established at S.C. Secerişul S.R.L. in
Pleniţa, located in Dolj county in the SW of Romania.
The plum accessions in the collection (71 of them original and 70 foreign ones)
belong to 13 species of the Prunus genus, plus 5 interspecific hybrids and 2 others.
The 141 accessions were phenotypically characterized using Bioversity
International (IPGRI), FAO and EURISCO descriptors.
The genetic resources present into this collection are valuable and can be
used into the future plum breeding programs for cultivars and rootstocks.

36
37
 ACKNOWLEDGMENT
Present work has been carried out with the help of project PN-II-PT-PCCA-
2013-4-1399 no. 168 in titled ”Conservation of native plum germplasm based on a
experimental model of molecular characterization and collection on farm”
(GERMPLUM).

REFERENCES
Botu M, 1999, Cercetări pentru stabilirea unor genotipuri şi soiuri valoroase de
prun pentru zona subcarpatică a Olteniei. Teză de doctorat, Univ. din Craiova
Botu M., Botu I., Preda Silvia. 2005, Metode de conservare a resurselor genetice
la speciile pomicole din genurile Prunus, Juglans şi Corylus. Lucr. Ştiinţifice "Prezent şi
Perspectivă în cercetarea pomicolă" Vol II, SCDP Constanţa
Botu M., Tomic L., Cvetkovic M., Gjamovski V. et al. 2012, Balkan Pomology -
Plums - Seed Net's WG for Fruit and Vitis, Exaktaprinting AB, Alnarp, Sweden
Butac Mădălina, Botu M., Militaru M., Budan S., 2011 – Romanian germplasm
fund and its use in plum breeding program, Scientific papers, R.I.F.G. Piteşti, Vol.
XXVII.
Botu I., Roman R., Turcu Elena, Botu M., Duţu I., 2001. Documentation,
characterization and evaluation of Prunus Genetic Resources of Romania. Project
Genres – 61 of the European Union Genetic Resources Programme 1467/94
Mehlenbacher S., Cociu V., Hough L.F., 1992. Apricots (Prunus) In: "Genetic
Resources of Temperate Fruits and Nut Crops. ISHS. Wageningen; 63-108
Ramming D.W., 1992, Plums (Prunus). In: "Genetic Resources of Temperate
Fruits and Nut Crops. ISHS. Wageningen: 233-258
xxx, 2001, Descriptors FAO/IPGRI For Genetic Resources, Roma
xxx, 2012, Descriptors for unploiding information from National Inventories to
Eurisco
xxx, 2016, FAO - Food and Agriculture Organization of the United Nations.
Statistics of Fruit Crop Trees (plum and apricot).

38
 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
 Food produce process ing
UNIVERSITY OF CRAIO VA
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

RESEARCH ON THE MULTIPLE WAYS FOR OBTAINING ECOLOGICAL

AND QUALITY SEEDS OF LAVANDULA
1*
Broșteanu Elena
1
University of Agronomic Sciences and Veterinary Medicine of Bucharest, Faculty of Horticulture
*Correspondence author. E-mail: [email protected]

Key words: Angustifolia, Ecological, Lavandula, Vera

ABSTRACT
This paper presents the influence of soil and planting density in Lavandula production
and seeds quality. The subsidiary objective was to obtain an ecological culture for high quality
seeds. The cultivars of Angustifolia and Lavandula ong Silver Mist have been studied at the
following planting distances: 0/25 with 71,428 pl ha, 70/30 with 47,613 pl/ha and 70/40 with
35,714 pl/ha. The setting up of the culture starts in the autumn of 2015, when the first row was
planted. The seeds production per plant increase direct proportionally with the planting distance.
In both varieties, the highest seed yield was obtained at a density of 71,428 plants per hectare.
In this case, the average seed yield per hectare was around 222 kg for Lavandula ong Silver
Mist and about 165 kg for the Angustifolia variety. The yield obtained is sensible higher, so
further quality investigation was performed for proving seeds quality.

INTRODUCTION
Lavandula angustifolia Mill which contains a volatile oil with a controlled
chemical composition and meets specific quality requirements. Our main goal
consisted in type of propagation, both generative, by seedlings and vegetative, by
rooted cuttings or by earthling up and separation of old shrubs. Species / subspecies /
cultivars of Lavandula in Romania, they were as follows:
 Lavadula Angustifolia Mill
 Lavandula Angustifolia ssp. Angustifolia.

MATERIAL AND METHODS

Determination of morpho-anatomical features of Lavandula species /
subspecies cultivated in Romania:
 evaluation of the volatile oil content extracted by hydrodistillation and
determining its chemical profile.
 production of seedlings from seeds
 by rooted cuttings
 separation of old shrubs

39
 RESULTS AND DISCUSSIONS
Lavandula is a mountainous, less demanding, sun-loving species. It is a
homogeneous, ethomophilous plant and, by generative propagation, it produces many
shoots with chemical morphogenetic variability and with diverse economic efficiency.
Lavender seeds are hard, covered by a poorly permeable coat, which leads to
long period needed to sprout and spring. After emergence, the lavender plant forms
shrubs as a result of growing new branches, increasing the flower production.
Lavender populations predominantly include forms which bloom once a year,
but other types bloom gradually or twice during vegetation. During the vegetation
period, lavender goes through the following stages:
- leaf-out
- budding
- blooming
- seed ageing
Table 1
Indices of quantitative features of lavender (2nd year of vegetation)

Number Plant Length of Length of Length of Number of

of height flower flower flower spike whirls on
versions (cm) (cm) stems (cm) (cm) flower spike
X X±Sx X±Sx X±Sx X±Sx
V1 48.5 27.2±2.4 20.4±1.2 6.8±1.3 6.3±1.1
V2 46.5 26.9±2.3 19.9±2.4 7.8±1.4 6.5±0.1
V3 47.0 25.7±2.2 18.8±1.5 6.9±1.4 6.8±0.8
V4 48.0 26.5±2.4 19.7±2.1 6.8±1.3 6.6±1.8
V5 54.0 31.5±1.5 23.5±2.2 8.0±0.7 7.7±0.5
V6 52.5 29.8±1.8 23.5,±2.3 8.1±1.3 7.4±1.0
VMt 41.5 24.5±1.9 17.4±1.4 6.3±1.1 6.0±0.5

Figure 1. Lavandula angustifolia plantation untreated.

The main factor that limits the spread of various lavender species is
temperature. Lavender plants begin vegetation at average daily air temperatures
around 10°C. The seed sown on the brink of winter directly in the field or in cold
40
beddings springs up in spring at 12-13°C provided it has proper humidity. Lavender is
a plant specific to dry biotopes and warm climate, with proper morphological features
that allow it to be drought tolerant. When cultivated on land with excessive moisture,
lavender suffers from rotting roots, which causes ageing and rapid drying of shrubs.
Lavender seeds sprout only when the surface soil layer they're in is moist enough.

Figure 2.Lavandula plantation after applying only physical treatment.

Moreover, in case of rooted cuttings, both for lavender and lavandin, moisture
is a decisive factor.
Final transplantation of rooted cuttings or seedlings will be done exclusively in
wet soil, in case of lack of moisture, plants will necessarily be watered at the planting
location.
Lavender and lavandin are light-loving species. Lavender plants which grow
under the sun always bloom at a higher rate, developing large shrubs, 3 times more
flowers. Light conditions the formation of leaves, as well as assimilation and production
of volatile oils. In order to increase the amount of light, lavender plantations are
recommended to be located in our country on land facing South or South-West.
The best results are obtained when lavender is grown on deep, limestone-rich,
permeable soils, with groundwater at depths of at least 2-2.5m.
Highly sandy soils, heavy, clay, cold and wet soils are not recommended for
lavender crops. Nitrogen plays an important role in the rejuvenation of shrubs, as it
stimulates growth, which is why young plants will be provided with more phosphorus in
the fertilizer mix and more nitrogen will sprayed on old plantations.
Growing seedlings from seeds. Sowing can be done both in late autumn,
October -November and in early spring, February-March, but only in specially prepared
beddings, which must be 1-1.2 m wide and 8-10 m long and even more. A 1-meter
space should be between beddings.
The seeds sprout in early spring when air temperature is constantly maintained
at 14-15°C. In case of spring sowing, the seeds germinate and sprout slowly, which
requires their 3-4-day moisturizing in water or their layering, which must start 1.5 - 2
months ahead. For this purpose, the seeds are mixed with wet river sand in a ratio of 1
: 5-6 and are kept out until sown, meanwhile being stirred to avoid molding. By the end
of the layering period, the seeds germinate and will be immediately sown in warm soil.
After the plants spring out of ground, when they have 2-3 pairs of leaves,
thinning is applied, leaving 3-5 cm space between plants. Until the end of vegetation, in
autumn, seedlings must be kept clean from weeds, by repeated weeding, must be
41
watered, fertilized and hoed. Concomitantly, at least twice, all the flower stems will be
cut to force the plant to grow branches as close to the ground as possible. The cutting
height of floral stems is approx. 8-10 cm above ground level.

Figure 3. Samples of Lavandula angustifolia seeds and yield.

The seedling is ready for transplantation, to the final location, at the end of
September and in early October when it is taken out, sorted and stored in layering in
sand or wet soil until planting.
Seedlings have the following characteristics:
– length of aerial part 12 cm
– length of main root 15 cm
– minimum 4 mm diameter and 3 branches
Growing seedlings by seeds is a cheap and fast way of propagating lavender.
Propagation by rooted cuttings: Cuttings are harvested autumn, in September-
October or in spring, March-April, during the vegetative rest period. Cuttings are
harvested from healthy, well-developed plants, aged 3-5 years. Cuttings are cut with a
very sharp knife or blade, under buds. Leaves are removed carefully to avoid
vegetative buds being destroyed. One wedge-shaped cut is done at the base of the
cutting, right after bud. The cuttings thus prepared have a length of approximately 5-6
cm and are ready for planting, which is to be carried out within 1-2 hours. In case of
longer storage, but not longer than one day, cuttings are kept in pots with clean water.
The land chosen for the beddings where cuttings will be planted must be
weed-free, located near a water source, but without risk of swamping. It will be
ploughed to 30 cm deep, leveled, shredded and then gently tamped, then seedbeds
will be arranged, allowing 40-50 cm space between them. The seedbeds shall have a
well sifted, 8-10 cm thick layer of sand, which, after leveling, is watered abundantly.
After watering, to facilitate planting, the next rows will be marked, using markers
specially prepared for this work. Planting is done manually, placing them with a 10.5
cm spacing. Immediately or concurrently with planting, the cuttings will be watered
abundantly, and the seedbed will be covered with window frames whose glass has
been previously varnished and covered with mats until the first roots emerge on
cuttings. When the first roots are formed, cuttings are strengthened by uncovering the

42
seedbed, for a few hours initially, exclusively during the morning or evening. When
floral stems appear, they will be cut to half to stimulate branching.

Figure 4. Different Lavadula variants and species, on the experimental plantation

CONCLUSIONS
Paper's general conclusions can be structured as follows:
A. The histoanatomical structure of the vegetative organs shows minor
differences between the two subspecies (L. angustifolia ssp. angustifolia / L.
angustifolia Mill), rather quantitative differences:
• Processed species, subspecies and cultivar
• Moment of the growing period and sampled plant part
• Location of origin • Climatic conditions during harvest
• Type of material (fresh / dry) subjected to processing.
B. The polyphenolic fraction varies qualitatively and quantitatively from:
• Processed species, subspecies and cultivar
• Climatic conditions during harvest
• Type of solvent used for extraction.
C. The antioxidant activity of the polyphenolic fraction varies with:
• Content of polyphenols present in extract, determined by species and
subspecies
• Cultivar, samples from Balteni (L. angustifolia Mill.) being more active than
cultivars collected from varieties cultivated in Dambovita.

ACKNOWLEDGMENT
Project's originality firstly derives from the investigation of certain new species
and varieties of Lavandula cultivated under pedo-climatic conditions specific to the
South-East of Romania. Another element of originality consists in total pharmaco-
botanical, chemical and biological issues used to explore the species under study. The
project contributes to wider phytochemical and correlation existing between
pedoclimatic conditions in our country and chemical composition, substantiating the
antimicrobial and antioxidant activity of species for Lavandula seed production.

43
 REFERENCES
Alexan M., Bojor O., Crăciun F., 1989, Medicinal flora of Romania. Vol. I, II.
Bucharest: Ceres Publishing House, pp. 118.
Ardelean A., Mohan Gh., Mohan R., 1998, Plants and health. Bucharest: Scaiul
Publishing House.
Bodea, C., 1982, Treaty of vegetal biology. Part II: Chemical composition of the
main crop plants. Romania: Acad. R.S. Publishing House, Vol. 4, pp. 105.
Cucu V., Bodea C., Cioacă C., 1996, Treaty of vegetal biochemistry. Part II:
Chemical composition of the main crop plants. Vol.2, pp. 76.
***, 1982, Medicinal and Aromatic Plants. Bucharest: Acad. R.S. Publishing
House, pp 408.
***1994, Bojor O. Aromatherapy, Bucharest, pp. 234

44
 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
UNIVERSITY OF CRAIO VA  Food produce process ing
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

THE CONCEPT OF ,,ROAD OF THE WINE” IN OLTENIA AND THE PLUS

OF VALUE FOR A TOURISTIC DESTINATION

Călugăru Iulian Viorel1, Giugea Nicolae1*, Mărăcineanu Liviu1, Berta Corina1,

1
Perţ Carmen
1
University of Craiova, Facultaty of Horticulture
* Correspondence author. E-mail: [email protected]

Key words: oenotourism, wine, road of the wine, touristic product, touristic potential, touristic
destination
ABSTRACT
The oenotourism is a recently phenomenon for Romania. In the european countries it
has been practiced for a few decades, spontaneous or organized. This expansion is determined
by the existence of two reasons fot the wine-growing tourism: on the one hand is the problem of
the development of the wine-growing field and on the other hand of a form of alternative tourism
to the traditional tourism.
In fact, the road of wine represents a well indicated and delimited touristic route in a
zone with wine-growing potential and with enough touristic attractions, natural and anthropical,
which can be used by the tourists, individual or in organized groups, in some periods of the year.
These roads, depending on their length, but also by the diversity of the objectives, can be seen
by the walking tourists or by bike, by car or even by carriage, in a day or in a few days.
The marketing of the touristic products includes the product of ,,touristic destination”. If
we compare it with other domains of activities, we observe that the tourism reacted with a certain
delay in the application of the methods and techniques af marketing. But, in our days, the
tourism and specially, the oenotourism in Oltenia is looking to extend and to go thoroughly into
these techniques in order to turn to good account the resources and to carry out a better
efficiency of the whole activity. The marketing is the process of an organization to be adopted to
the market in a creative, productive and profitable manner.

INTRODUCTION
The european countries with a long tradtion in wine-growing production,
understood a long tine ago that the wine is not only a liquid in a bottle, by the contrary
the wine is a sory, a history, a legend and a civilization, it is a rural tradition and an
urban charm. They understood that is very important for the wine lovers to enjoy the
famous wines in the restaurants or at home but in the same time, to discover the
universe of the vineyard, the place where it grows – in the wonderful valleys or on the
sunny and majestic hills in the mysteriously Oltenia.
The oenotourism and the ecotourism appeared first like an opportunity in
affairs, which could be a succes on a long term only if it is a responsiable affair, which
respects the market rules and the lasting development in the same time.
Establishing the nair objectives of this study, we proposed to ourselves to
identify the factors which can produce the plus of value for a wine-growing touristic
destination. In time, the travel reasons of people become very diverse, appearng new

45
needs and whishes, multiple and more complex, caused by the changes at the level of
society.
More than a century ago, nobody thought to pay a visit in a vineyard. The
oenotouristic motivation (vocation, inclination) which generates the touristic request is
always ery personal, subjective, determined by the psychological impetuses
(influenced by the environment).
In the social plan, the tourism (generally) and the oenotourism (specially) have
a substantial contribution to the raising of the level of education, culture and civilization
of the people. Stimulating the exchange of values, the tourism encourages the
enrichment of the cultural horizon for the tourists and for the local population.

MATERIAL AND METHODS

The touristic destination represents the place or the geographical space where
a visitor or a tourist stay for a night or for a longer perios of time, or the final point in the
vacantion of the tourists, no matter if they travel for touristic or for affair reasons
(Stăncioiu, A. F., 1999).
The method used in this study was the geographic method, which represents a
complex, synthetic approach of the tourism fenomenon in the territory, starting from the
existence of potential and built touristic landmarks and from the touristic phenomenon
in its complexity, and also from the fact that tourism evolves in the environment, more
or less humanized, which is the “raw material” for this and with whom is in close
interaction.
Geographic method involves, as a matter of fact, the use of the principles of
the distribution in an surface (or area) and geographic integration, specific to
geography as a science.
According to this method all information about tourism activities offered by
specialists in this field, in relation to the specialization of each, are dispersed, but the
overview of the touristic phenomenon results only in the systematization of knowledge
and their geographic integration into a territorial and geographis functional system.
It requires a special (expansion, shape, complexity) and positional
(geographic) approach of the objectives and of the tourism phenomenon in general, in
relation to the areas and forms of the nearness tourism (or at a higher territorial level),
and functional and territorial integration of the touristic activity; a causal approach of
the touristic phenomenon, but also an ecological and socio-economic approach
through the relationship with the environment and human communities (Glăvan V.,
2005).

RESULTS AND DISCUSSIONS

The main objective of this research was the identification of some wine-
growing trouristic destination in Oltenia, as well as the factors which can produce a
plus value of these destination. The wine-growing tourism shortens, in fact, the
distances between the wine lovers and the producers, while the tourists who can see
live the process of wine producing and the necessary effort of this process will be the
first promoters of the consumation of good and authentic wine.
The wine is a real part of the european identity, that's why is very important to
maintain this common cultural patrimony in its whole diversity, and to kepp this cultural
inheritance for the next generation.
The wine-growing in Romania is now in a period of revival, is in full swing.
Romanians begin to redescover the wines, the producers become more and
more experience and wines of a good quality are produced in our country, too.

46
Although in spite of these trumps, the connection between wine-growing aand tourism
is weak represented.
In Oltenia the wine-growing tourism is in an evidently development, being
helped by the investments carried out by the wine cellars, as well as by the request
from the costumers for this kind of travels, directed to same experiences. The potential
of development is great the wine lovers being attracted by the visits in vineyards and
wine cellars by the conversations with the experts in wine-growing and not finally, by
the discovered in Oltenia. More than these the old mansions near the vineyards whide
have been recpnditioned and changed into units for accommodation are points of
attraction with their legends for the Romanians and foreign tourists. Until recent time
the wine lovers in our country werw forced to travel abroad in order to practice the
wine-growing tourism, but in our days they can do this type of tourism just in our
coutry. The geographical position, landscape, diversity, history and the native varieties
of wines are clear advantages which can be used by the owner of wine cellars in order
to attract the Romanian and foreign tourists to the local wine-growing zonees.
The wine cellars in Oltenia can be visited all year round, but the most attractive
landscapes are in the period between april and october
Now, we small desribe four wine-growing destinations in Oltenia, which can be
included in a concept of “road of wine” in the north part of Oltenia.
1. The wine cellar ,,AVINCIS” is situated in Vâlcea District, on the Hills of
Drăgăşani
(Drăgăşani town is situated at a distance of 6 km far fromn wine cellar), at abot
200 km dar from Bucharest (two hours and a half by car).
ANVICIS is the symbol of the rival of the Romanian wines, uder the sign of
tradition and modernism. This adventure started by the discovery of one passion in
2007. When Cristina Stoica ame back with his husband, Valeriu, in the brithplace of
their family, in the region Drăgăşani. So they started their project to give a new life to
the family domain, in order to give back the older charm of the vineyard from the eng of
the place for the tourists who like the beautiful and the good wine.
In order to build for a long time the personality of their vineyard, Valeriu and
Cristina Stoica tried to use the knowledges of the young and talented French oenolog
Ghislain Moritz, graduate of the Dijon University, who had experience in wine-making,
working in Bourgone region and in Portugal. This vineyard with 50 hectares ( 41
hectares active ) occupies the whole hill and its slow slopes until the valleys. Out of the
old varieties, today are planted “Tămâioasa Românească”, “Negru de Drăgăşani” and
“Crâmpoşie Selecţionată” and besides all these other varieties with a foreign origin
which have benn introduced in Drăgăşani wine-growing. So that, varieties as
Sauvignon Blane, Pinot Noir, Cabernet Sayvignon or Muscat Ottonel have adapted
very well at the climate and soil condition. AVINCIS Wine Cellar from the “Domeniul
Vila Dobruşa” offers an unique experience. With a capacity of accommodation for 28
persons in double system or 14 persons in simple system, AVINCIS offers to its visitors
10 modern double rooms complete equipped and 3 flats situated above the wine cellar,
with a splendid view about the domain and vineyard.
AVINCIS wine cellar is surrounded by touristic objectives, Dobruşa Villa being
an important point of attraction in the region. Dobruşa hermitage, the monasteries of
Oltenia ( In One-Wood, Horezu, Arnota and the fortified houses from Măldăreşti ) are
only a part of the beautiful of this zone.
2.BAUER WINE CELLAR – founded in 2012 is situated in Drăgăşani , Vâlcea District,
in the middle of the wine-growing Drăgăşani , wine-growing zne Oltenia. This new wine
cellar is conceived to permit the process of wine-marking for a small quantity of grapes
out of rare or unfound varieties yet by the wine lovers, gathered in the old and well
47
carefully lots. The philosophy of the wine cellar “Natural Creativity” is the mirror of
confidence in diversity, not in the concept of “the best wine”. Varieties of grapes:
sauvignonasse, sauvignon Blanc, Crâmpoşie Selecţionată, Cabernet Sauvignon,
Merlot, Petit Verdot, Fetească Neagră.

Figure 1 AVINCIS wine cellar (http://www.revino.ro)

Drăgăşani wine-growing is rich in traditional varieties of wine, in natural

beautiful and diversity, smat people, exceptional experts in wine-making and wines
with personality. Bauer wine cellar can be visited every seasin. It is waiting for you to
taste its wines in winter, spring, autumn or summer. Preious reservation. Grups
between 4 and 18 persons.
3. Ştirbey Wine Cellar – is situated in Drăgăşani, Dealul Olt Street, Vâlcea
District. The owners are Ileana and Jakob Kripp and wine-making Oliver Bauer.
Since more than 300 years, the princely family Ştirbey had a wine-growing
domain in Drăgăşani. The wines of thw princely family Ştirbey are produced according
to some principles developed and kept from generation to generation. Here they
cultivate traditional varieties of grapes, adapted and with a good evolution in time at the
soil and climate conditions of this vineyard. These varieties of grapes of wine-making,
so that are produced only wines from pure varieties in order to emphasize the
characteristics of every variety.
Soil, climate, flora and fauna, but also the traditions of people who work in the
vineyard influences very much the evolution of wines and grapes, giving to every lot of
vineyard another characteristic which can be synthetized as Trroir or, in an universal
meaning, as Genius Loci – the spirit of the place.
The owners want to refind this Genius Loci in the wines produced and sold
here.
They are sure that the forces of nature give us products healther and more
lasting than any teachnical interention. The vines have a good care, with strict manual
works, so that the chimical treatments are reduces to the minimum. In the process of
wine-making is followed the clock of the nature fot the fermentation and maturising,
and it is not forced the process by the technical interventions.
4. Drăgăşani Domain – is situated in Drăgăşani, Dealul Viilor Street, Vâlcea
District. Situated on Dealul Viilor, in Drăgaşi Domain and its four points of attraction –
the wine-growing, wine cellar, the saloon for wine tasting and the hotel – in an
ensemble which proposes to itself to leads the guests to refind the nature and their
own aspiration. Thia pilgrimage is influemed by the panoramic view about the wine-
growing, afforested hills and flowing Olt, but also by the meeting of the people with
their stories
48
 Very interesting are the descriptions of some wines : “Pinot Gris 2015”. You
Suddenly realise that you are in a flowers filed. You don't scare yourself, such
experiences have the tendeny rather to calm you. You raise your head between
poppies and can see yellow in your glass. Or is the yellow straw of the field? Yhe fresh
aroma make you to feel cool, the air has a semidry aspect, nothing hurry you,
everyehere there is an unmotion as if you keep your breath. “Sauvignon Blanc 2015”.
You try to understand words which help you to describe what you drik. And in your
mind appear: poems from the seventh from, love letters fom the secondary school, an
acacia forest, citric fruit in an castern market, the balance of the ships in the sea, a
shell in the sand. Finalu, the fresh taste shakes you and you can seen another
imagine-you, the glass, the hat for sun. “Merlot 2015” the cork has a smell as the
phrases like all the time in the world. The intense red and violet remins you about a
storm from your childhood, when the sky wew full of fire, and the rain were
extingnishing it. The aroma of blackberrues, currants, cherries and spices determine
you to ask yourself if you are on a verandah or in a forest. The balanced but delicious
taste needs some cheeses and closed eyes etc.

Figure 2. Drăgaşi wine cellar

(http://www.domeniul-dragasi.ro/)
Limits. In Oltenia there are many wine cellars wines growing, history, touristic
objectives and very well roads. But in the same time, we have no enough rooms for
accommodation and we new marketing which could promote these wine growing
destinations. We haven't structure and coherentness. ,,Three principle condition the
succes of the promotion, and these are the three << C >> : continuation, convergence,
co-ordination “ ( Llanguar R., Hollier R., 2001)

CONCLUSIONS
The oenotourism in Oltenia must constitute the subject of the project
management. The road of wine has to become a touristic route, with wine-growing
products, with points for wine-growing products, with points for wine tasting and
touristic, cultural and natural objectives which can be developed by well made
programmes from one until three days for the people fond of tourism. The multitude of
resources / types of resources doesn't facilitate implicit the development of the tourism
in a certain region. By the contrary, it needs intense efforts of marketing, The new
marketin initiates, negotiates and leads favourable relations for change with the main
groups of interest, in order to turn to good account the potential of these relations and
to create the fidelity with the brand of the products, because and services not only for
products and sevices not only for their use value to spend his free time, but also for
their value of social connection. The geographical position, landscapes, diversity,
history and the native varieties of grapes are real advantages and the wine cellars
owners can use them in order to attract the Romanian and foreign tourists in the local
wine-growing zones.
49
 The oenotourism has a favourable inflluence about other national economy
domains. Consequently it is necessary to be well known the wine-growing destinations
in Oltenia, because they can create the concept of “road of wine” in Oltenia and, in the
same time they can create a plus of value for a touristic destination with the trumps of
the wine cellars in Oltenia.

REFERENCES
Glăvan V., 2005, Geografia turismului, Editura Fundaţiei România de Mâine,
Bucureşti.
Hollier R., Lanquar R.,2001, Le marketing touristique, Editeur Presses
Universitaires de France - P.U.F.
Stăncioiu A. F., 1999., Dictionar de terminologie turistică, Editura Economică,
Bucuresti.
***http://www.revino.ro/turism-viticol/crame/avincis-vila-dobrusa-crama-restaurant-
hotel-cazare-vin-vizite-degustari-evenimente-dragasani-cramposie-valeriustoica.
Html
***http://www.revino.ro/turism-viticol/crame/domeniul-dragasi-crama-cazare-
dragasani.html#prettyPhotoăgallery]/0/

50
 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
 Food produce process ing
UNIVERSITY OF CRAIO VA
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

THE OENO-TURISM A CONSTANT FORM OF TOURISM IN OLTENIA

1 1* 1 1
Călugăru Iulian Viorel , Giugea Nicolae , Berta Corina , Carmen Perţ ,
Mărăcineanu Liviu1
1
University of Craiova, Faculty of Horticulture
* Correspondence author. E-mail: [email protected]

Key words: oenotourism, wine, touristic product, sustainable development, wineries

ABSTRACT
The development of the tourism industry has recently been displayed through its
returning to nature and original cultural values. The oeno-tourism is one of the most valuable
forms of tourism. These forms have as the main purpose the preserving of the environment with
a high attention to the tourist’ education regarding the protection and the preserving of the
environment.
The connection between the tourism and the environment has a high importance
because they are the most important conditions to be fulfilled in order to develop the tourism.The
touristic development of the population, the variety of the motivations cand lead to a single great
contribution designed to stop the damage of the environment caused by different activities, thus,
through some rules, which must be taken, to fulfill the protection and the preserving of the
environment. Any dangerous action can lead to transformation and, most of the times, is an
injury to the environment: the decay and the defeat of its resources also of the economical
balance and all these can lead to the damage of the health and of the existence of the next
generations.Generally speaking, there are objective factors (when natural phenomena are
unfavorable displayed) and subjective factors, which are caused by human activities, that affects
and change the environment. Through different activities (controlled and uncontrolled tourism),
Man has a great contribution in the natural destruction. Those who are involved in oeno-touristic
activities are aware of the effects on the touristic development, of their impact on people’s
activities.

INTRODUCTION
Even though the tourism, at its beginning, represented a growing economical
factor for protecting and preerving the environment, nowadays the ecological balance
and the natural resources balance is highly affected, mainly through the growth of the
tourists flood. This is why it is neccessary to reorganise the area, to adapt to the needs
of the tourists, to support them, to reorganise the space, and this is a complex duty.
A constant tourism means an industry which tries to have the smallest impact
on the environment and on the local culture, but, in the same time, tries to support the
incomes, to diminish the unemployment and to support the local eco-systems.
The constant development must become one which fulfills all the needs
without affecting the future generations‟ abilities. An uncontrolled tourism can lead to
the development of the environment but also to its damage. According to J.Krippendorf
51
„ if we can lose and then rebuild the economical side of a country, when talking about
tourism, where the essence is the Landscape and the Earth, once the balance lost, it
stays lost forever.”

MATERIAL AND METHODS

The touristic destination represents the place or the geographical space where
a visitor or a tourist stay for a night or for a longer perios of time, or the final point in the
vacantion of the tourists, no matter if they travel for touristic or for affair reasons.
The method used in this study was the geographic method, which represents a
complex, synthetic approach of the tourism fenomenon in the territory, starting from the
existence of potential and built touristic landmarks and from the touristic phenomenon
in its complexity, and also from the fact that tourism evolves in the environment, more
or less humanized, which is the “raw material” for this and with whom is in close
interaction.
Geographic method involves, as a matter of fact, the use of the principles of
the distribution in an surface (or area) and geographic integration, specific to
geography as a science.
According to this method all information about tourism activities offered by
specialists in this field, in relation to the specialization of each, are dispersed, but the
overview of the touristic phenomenon results only in the systematization of knowledge
and their geographic integration into a territorial and geographis functional system.
It requires a special (expansion, shape, complexity) and positional
(geographic) approach of the objectives and of the tourism phenomenon in general, in
relation to the areas and forms of the nearness tourism (or at a higher territorial level),
and functional and territorial integration of the touristic activity; a causal approach of
the touristic phenomenon, but also an ecological and socio-economic approach
through the relationship with the environment and human communities. ( Glăvan V.,
2005).

RESULTS AND DISCUSSIONS

The major aim of this research was to identify the traits of the oeno-tourism in
order to become a developed one. Also, we wanted to establish some objectives for an
everlasting development of the wine-making industry in Oltenia.
A constant development means economic efficiency, social and cultural
cohesion, protection of the environment.(Glăvan V., 2005). The development of the
oeno-tourism is not automatic because it is connected to image and reputation.
The wine making tourism in Oltenia represents an important opportunity of
marketing and a possibility to develop the inside tourism. The oeno-tourism in Oltenia
is a method to promote the wine-making industry including a physical, social and
cultural dimension of the wine-making scenery.
The oeno-tourism in Oltenia fulfills some conditions and becomes a constant
one:
1.Supports the place integrity: the vineries from Oltenia emphasise the local
architectural trait of the area by keeping the local architectural motifs and by melting
the local culture with its chime, the using of specific materials, the local cuisine, natural
attractions and landmarks, aesthetics. The profits from this form of tourism lead to the
growth of services.
52
 2.Awakes the quality not the quantity: the vineries owners do not measure the
success through their visitors but through the time spent there by the visitors, the
expenses and the quality of the experience.
3.Do not abuse the product: The pressure growth is anticipated and applies
management limits and techniques. Thus, the groups of visitors are always small. The
firms must work to support the natural and cultural resources. Visiting the historical,
artistic and architectural landmarks in bad conditions (candles use, lack of technical
appliances) led to the damage of important and valuable pieces of art.
4.It is informative: the traveler does not only find information about their
destination, but also finds about how to help and support, thus, the experience
becomes memorable. By participating at some specific activities, the tourists will learn
interesting things. The festive traditions and customs usually represents the end of the
wine-making experience which can be in the same time with a religious holiday( The
Grapes Dance, The Vineyard Baptism).
5.Supports the local people: the vineries owners try to hire and to train local
people, to buy local products and to use local services.
6.Preserves the resources: The ecological aware tourists help the pollution
diminish, the use of electric power, the use of water and chemical products.
7.Values the local cultural traditions: the foreign visitors observe and learn the
traditions and the local customs. Most vineries include artistic events with folk music
and dances. Thus, the firms are likely to practice the wine-making tourism to fulfill the
tourists‟ requirements.
8.Means awesome holidays: the visitors will take with them interesting
information and they will send their friends for the same experience, for sure, which will
assure a constant flood of tourists.
Regarding the development rules, the building of equipment and the touristic
wine-making appliances in Oltenia are fulfilled through: lower buildings with an
attractive image, sceneray traits, parking places, natural resouces. (Snak O. and coll.,
2001)
The constant oeno-tourism helps the understanding of the touristic activities on
the natural , cultural and human environment and assures a touristic development.
The wine-making tourism in Oltenis has the following aims:
-To influence an to motivate the travelling
-To assess the positive results of the past
-To get an open attitude in the relations with the employees
-To practice an active education in order to protect the natural and cultural
resources of the touristic industry in order to understand the environmental problems.
Limits. It is necessary to extend a complex research regarding the constant
development of the tourism, generally speaking, and of the wine-making tourism. The
economic efficacy, the growth of the social cohesion and the protection of the
environment must be considered.

CONCLUSIONS
Like any other human activity, the oeno-tourism needs touristic resources and
participates at the damage and the pollution of the environment.

53
 The oeno-tourism of our area needs a healthy environment with beautiful
landscapes as a starting-point for all the other touristic activities. The wine-making
tourism of this area represents an opportunity for all the wine-producers and also a
way to develop the inside tourism.

REFERENCES
Glăvan V., 2005, Geografia turismului, Editura Fundaţiei România de Mâine,
Bucureşti, pag. 40-42, 191.
Snak O., Baron P., Neacşu N., 2001, Economia turismului, Editura Expert,
Bucureşti, pag 468.

54
 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
 Food produce process ing
UNIVERSITY OF CRAIO VA
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

DETERMINATION OF FACTORS THAT REDUCE THE POTENTIAL OF

DEGRADATION OF VEGETABLE OILS

Ramona Căpruciu1*
1
University of Craiova, Str. A.I Cuza, No. 13, Craiova
Corresponding author: E-mail address: [email protected]

Key words: cold and hot pressed sunflower oil, quality.

ABSTRACT
In this study, it is desirable to establish a comparative frame in terms of the degree of
oxidation between sun-pressed sunflower oil and sun-pressed sunflower oil obtained from seed
lots with a different moisture content. The use of these categories of oil was aimed at obtaining
data on their physicochemical stability, organoleptic analysis and the main quality indices (acidity
index, peroxide index), determinations with a precise indication of the potential for degradation of
the studied oils.

INTRODUCTION
In the food industry, the vegetable oil sector occupies an important place by
supplying the population with essential products (edible oils, margarines, mayonnaise,
etc.) by Gunstone et al, 2011, Popa, V.M et al, 2010.
In order to obtain a good quality oil, the raw material must correspond to the
qualitative parameters included in the standards in terms of both the oil content and the
moisture content, the parameters being maintained only if optimum conditions are
ensured in the storage spaces (M.M. Duda et al. 2007, Sarca Gh.,2004).
Sunflower, as a source of vegetable oil, occupies the first place in our country.
High content of unsaturated fatty acids, predominantly linoleic acid (44-75%) and oleic
acid (14-43%) and the presence of low linolenic acid (0.2%) give the high nutritional
value of the sunflower oil (O'Brien R., 2008). The growing demands of the Romanian
market for raw oils obtained by cold pressing are proven by the increasing desire of
consumers to know as many technological, nutritional, medical aspects as these oils
(D. Mozaffarian et al, 2009, Căpruciu R. 2011). They are also tasteful and contain
plenty of protein and liposoluble vitamins.

MATERIALS AND METHODS

Determination of humidity content of oilseeds The determination shall be made
within 16 hours of receipt of the laboratory sample in closed, labeled packages. It dries
in the oven, in a stationary atmosphere. Samples of sunflower seeds are pre-crushed by a
laboratory mist and then passed through a 3.15 mm sieve. The screening sample is
homogenized and introduced into two troughs, which are fed into the oven together
with the lids, for the duration indicated in Stas for each oleaginous feedstockThe time
and temperature for sunflower seeds was set at 130 +/- 30°C, 1h. After the drying

55
process ended, the vials were capped and immediately placed in the desiccator for 30
minutes. Once removed from the desiccator, they are weighed again. Calculation is
performed according to the formula and is expressed in% umidity.
Determination of organoleptic examination for established oil samples By
performing the organoleptic examination according to STAS, the subjective
examination of the color, odor, taste and appearance qualities is done. Determination
of appearance consists in visual observation in glass container (beaker Berzelius) in
natural light. Analyze the clarity, presence of suspensions, flocculation or sediment.
Color determination is made by examining the (natural) light of oil samples transported
in a colorless glass container, noting whether their color corresponds to the standard
and quality. Color change can occur due to certain processes such as wetting, heating,
milling or inappropriate drying of the raw material. Examination of smell is done
directly, by inspirational evidence. The olfactory test determines possible defects: stale
smell (taken from inappropriately stored seed), altered, moldy, rancid, fermented,
infestation (eg honey odor to infestation with mites of the raw material from which the
oil originates). Determination of taste is made by chewing of 5-10 ml of oil. Before and
after each determination rinse the mouth with water. It is appreciated that the taste
meets the standard by assessing whether the oil has a chemical taste or other foreign
taste. Expression of the results is made by entering in the quality document any
reported defects.
Determination of acidity index The acidity index is the amount of KOH, in mg,
required to neutralize free fatty acids from one gram of fat. Titrate the dissolved oil in
ethanol with 0.1N NaOH solution in the presence of phenolphthalein. Samples of liquid,
homogeneous and clear fluid at room temperature are analyzed as such. Samples of
liquid oils at room temperature, with insoluble waxes or glycerides, are heated to about
60 ° C, homogenised and filtered through filter paper. Weigh 5 g of the sample
(sunflower oil) - (depending on the acidity of the sample) to the nearest 0,01 g and
pass it quantitatively into an Erlenmayer glass. Over 100 cm3 of solvent is placed over the
sample. It is then agitaten until complete dissolution of the sample and add 5 drops of
indicator. It is then titrated with sodium hydroxide solution to the point of turning of the
color (pink in the case of phenolphthalein). Calculation is performed according to the
formula and is expressed in % oleic acid.
Determination of peroxide value The peroxide value is used in addition to other
quality parameters, for determining the oxidation degree of a product which is
composed of fat. This index is a measure unit for the oxygen content linked with
peroxide in oils and fats and particularly for the hydroperoxides. The peroxide value
was measured by determining the amount of converted iodide to iodine under the
action of the active oxygen of the peroxide.
The result was expressed as the number of milligrams of active O2/kg of fat.
There is only one principle for both methods of the peroxide value determination: the
oxidation of iodides to iodine through the active oxygen of peroxide, and measuring the
amount of free iodine by titration with 0.01 N sodium thiosulfate solution. An index
expressed in milligrams of active O2 higher than 20, indicates that fats are rancid. If this
index is very low, it has no meaning, in this case a simple heating at 130°C is sufficient
to destroy the peroxides (Căpruciu R. 2016).

RESULTS AND DISCUSSIONS

In order to determine which of the two assortments of oil (obtained by hot
pressing from seeds with normal humidity (8%) and high-moisture seeds (17%) and
obtained by cold pressing from seeds with normal humidity (8% high-grade seeds
(17%) retain their high-quality qualitative factors during storage, physico-chemical and
56
organoleptic analyzes were carried out on both raw material, sunflower seeds and oil
samples obtained.
For the steps in which the quality of the oils taken in the assay was
established, it began with the organoleptic analysis immediately after production, 14
days after production and 30 days after production. Determination of oil moisture is a
quality analysis in the context of obtaining oil samples required for the study (Table 1).

Table 1
Determination of humidity on the lots of sunflower seed studied

Component Sunflower seeds

Lot 1 Lot 2
Humidity (%) 8 17

As a result of this determination, the two seed lots have a different moisture
content, making it interesting to note whether this aspect influences the quality of the
finished product or not.
After the determination of the humidity, laboratory samples were developed for
the analysis of the quality factors. Thus, from each batch of seeds, two samples were
formed in order to obtain by pressing (cold or hot) the raw oil. Samples were scored as
follows: UR1 - raw oil obtained by cold pressing the seeds of lot 1 (8% humidity); UR2 -
raw oil obtained by cold pressing of seed in lot 2 (17% humidity); UC1 - raw oil
obtained by hot pressing of seeds in lot 1 (8% humidity); UC2 - raw oil obtained by hot
pressing of seed in lot 2 (17% humidity)
The organoleptic analysis was carried out under normal laboratory conditions
at a constant temperature of 20 °C under natural light conditions, the data obtained
being shown in Tables 2 and 3. The overall aspect of the studied samples is oily mass
with a high degree of shake suspension in the cold-pressed sun-float oil and with an
average stirring suspension in the hot-pressed sunflower oil. Also, when tasting, both
cold pressed sunflower oil and hot-pressed sunflower oil showed intense flavor specific
to the raw raw material from which it was obtained. The UR2 sample is slightly bitter.
For the odor analysis, rapid inhalation from coated pots for a few seconds was
performed, the four samples showing a characteristic odor of the raw material from
which they originated, more pronouncedly perceptible to sun-dried sunflower oil
obtained by hot pressing. The color of the analyzed samples was more intense for the
oil obtained by cold pressing. Samples obtained from high moisture seeds showed
spots or grains in the oil mass. By comparison organoleptically, the analyzed samples
reveal differences in color, consistency, smell and taste, differences in raw material
moisture and method of production (cold or hot pressing).
The oil samples taken in the study were analyzed qualitatively by two physico-
chemical indices recorded in STAS: the acidity index and the peroxide index, the main
indices giving clear data regarding the quality of the obtained oil, especially their
oxidative potential in storage time. Laboratory analyzes reveal differences between the
two categories of analyzed oils (cold and hot pressed) in terms of the value of the
physical and chemical indices analyzed, the differences being noted even within the
same type of oil (different lots table 4). It is noted that the free acidity expressed in%
oleic acid in sunflower oil obtained by hot pressing is lower than that recorded in the
cold-pressed sunflower oil. Also, samples of sunflower seed with low moisture content
show a lower acidity, maintaining the ratio during the analysis (30 days after
production).
57
58
 Table 4
Determination of the acidity index (I.A) in the studied oil types

Oil types
Storage time Sunflower cold Sunflower hot
pressing pressing
UR1 UR2 UC1 UC2
Acidity index (I.A) At production 1,1 1,4 0,5 0,9
(%oleic acid)
14 days 1,4 1,9 0,7 1,4
30 days 1,8 2,3 1,1 1,8

In which: I.A. – the acidity index; UR1 - raw oil obtained by cold pressing the
seeds of lot 1 (8% humidity); UR2 - raw oil obtained by cold pressing of seed in lot 2
(17% humidity); UC1 - raw oil obtained by hot pressing of seeds in lot 1 (8% humidity);
UC2 - raw oil obtained by hot pressing of seed in lot 2 (17% humidity)
Table 4 shows that the acidity index records double values of acidity for
sample UR1 versus UC1 immediately after production (1.1% versus 0.5%). Two weeks
after production, all samples analyzed showed an increase in acidity, after one month
of production and storage under the appropriate conditions (room without light, aerated
at 8 °C) only the sample UC1 (raw oil obtained by hot pressing of the seeds in lot 1
(8% humidity) being fit for consumption, according to recorded data. Due to the
technological process of production which involves keeping oil samples in contact with
the air for a longer period of time (compared to the production of refined oil), both
during the actual pressing and during the decanting, there may be oil degradations
visible to the eye free (especially color change but also smell and taste).
For this reason, the determination of the peroxide index (Table 5) together with
a thoroughly organized organoleptic analysis can lead to the exact determination of the
oxidative potential of the analyzed samples.
After determining the peroxide index, it is found that the cold pressed oil
samples degrade faster than the hot-pressed oil samples. Also, oil samples obtained
from seed with improper humidity show upward values (figure 1) of the peroxide index
over a period of 30 days, which means that these oils are not suitable for storage.
Following the physico-chemical analysis of the oil samples analyzed,
recommendations can be made regarding consumption and storage period.
Table 5
Determination of peroxide index (Ip) in studied oil types

Oil types
Sunflower cold Sunflower hot
The peroxide index Storage time pressing pressing
(Ip) UR1 UR2 UC1 UC2
(meq O2/Kg) At production 3,41 3,88 3,22 3,64
14 days 3,66 4,20 3,65 3,88
30 days 4,20 5,80 4,14 4,23

In which: Ip- the peroxide index; UR1 - raw oil obtained by cold pressing the
seeds of lot 1 (8% humidity); UR2 - raw oil obtained by cold pressing of seed in lot 2

59
(17% humidity); UC1 - raw oil obtained by hot pressing of seeds in lot 1 (8% humidity);
UC2 - raw oil obtained by hot pressing of seed in lot 2 (17% humidity)
Thus, out of the four analyzed samples, sunflower oil obtained from seeds
with normal humidity by hot pressing is recommended for consumption.

Figure 1 Determination of peroxide index (Ip) in studied oil types

It should be noted that the indices obtained for samples from seeds with
normal humidity but especially for samples of high humidity seeds indicate that they
are not suitable for long-term storage and are recommended to be used within a
maximum of two weeks (for sample UR1) and within 1 week (for UR2) as soon as the
oil has been in contact with the air (desintegration of the container) and keeping it at 8-
10 ° C.

CONCLUSIONS
Following the determination of moisture, laboratory samples were developed
for the analysis of the quality factors, from each seed lot being taken two samples to
obtain by pressing (cold or hot) the crude oil.
By comparison organoleptically, the analyzed samples reveal differences in
color, consistency, smell and taste, differences due to the humidity of the raw material
and the method of production (cold or hot pressing).
Laboratory analyzes revealed differences between the two categories of
analyzed oils (cold and hot pressed) in terms of the value of the physico-chemical
indices analyzed, the differences being noted even within the same type of oil (lots
diferent).
The free acidity expressed in% oleic acid in sun-dried sunflower oil is lower
than that recorded in cold-pressed sunflower oil. Samples obtained from sunflower
seeds with a low moisture content have a lower acidity, maintaining the ratio during the
60
analysis period (30 days after production). As a result, oil samples obtained from high
moisture seeds are not suitable for long-term storage. In order to reduce the oxidative
potential of these samples it is recommended that the seeds be subjected to the drying
operation with the normalization of moisture before the pressing process. The study
concludes that the moisture content of sunflower seeds is a first factor that can
increase the potential for degradation of the oils analyzed, this aspect being of interest
in keeping the oil in the household after the container has been unsealed and the oil is
kept at a temperature room or higher (cooking temperature during cooking) or if the
batch was not purchased and is nearing the end of the shelf life.
Air, by the presence of oxygen, is another factor that can lead to the oxidative
depreciation of the analyzed oil samples, requiring shortening the contact time
between the oil and air mass as a reduction factor of the oxidative potential.
Determination of the peroxide index along with a thoroughly organized organoleptic
analysis could lead to the exact determination of the oxidative potential of the analyzed
samples. Following the physico-chemical analyzes of analyzed oil samples, sunflower
oil obtained from seeds with normal moisture by hot pressing is recommended for
consumption.
It should be noted that the indices obtained for samples from seeds with
normal humidity but especially for samples of high humidity seeds indicate that they
are not suitable for long-term storage and are recommended to be used within a
maximum of two weeks (for sample UR1) and within 1 week (for UR2) as soon as the
oil has been in contact with the air (unsealing of the container) and keeping it at low
temperatures.

REFERENCES
Căpruciu Ramona, 2011, Study of sunflower oil quality under different conditions
of storage. Lucrare Ştiinţifică. Analele Universităţii din Craiova. pp.83-97
Ramona Căpruciu, 2016, Metode de analiză și control în industria uleiului. Ed.
Sitech. Craiova
M.M. Duda Şi A. Timar, 2007, Condiţionarea şi păstrarea produselor agricole.
Editura Academică Pres. Cluj-Napoca
Gunstone, Frank, D., 2011, Vegetable Oils in Food Technology: Composition,
Properties and Uses, Second edition Wiley Blackwell Publishing
Ana Leahu, Mircea Oroian, Sorina Ropciuc, 2007, The quality and stability of
walnut oil under the influence of storage conditions. USAMV Iași, Nr. 65(21)
Martínez, M. L., Penci, M. C., Ixtaina, V., Ribotta, P. D., & Maestri, D., 2013, Effect
of natural and synthetic antioxidants on the oxidative stability of walnut oil under
different storage conditions. LWT-Food Science and Technology, 51(1), 44-50
D. Mozaffarian, R. Clarke, 2009, Quantitative effects on cardiovascular risk
factors and coronary heart disease risk of replacing partially hydrogenated vegetable
oils with other fats and oils, European Journal of Clinical Nutrition 63, S22–S33;
doi:10.1038/sj.ejcn.1602976
O'Brien R., 2008, Fats and oils,: formulating and processing for applications. CRC
Press,.680 p
Popa, V.M., Hadaruga, N.G., Daniel I. Hadaruga, I.D., Gruia, A., Raba, D.N.,
Moldovan, C., Poiana, A.M., 2010, Fatty acids composition of some vegetable oils

61
obtained in the west area of Romania, Journal of Agroalimentary Processes and
Technologies,16,(3), 394-398
Sarca Gh.,2004, Materii prime vegetale. Depozitare și păstrare. Editura
Universitați din Oradea.

62
 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
 Food produce process ing
UNIVERSITY OF CRAIO VA
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

QUALITATIVE ANALYSIS OF TYPES OF ICE CREAM

Ramona Căpruciu1*
1
University of Craiova, Str. A.I Cuza, No. 13, Craiova
Corresponding author: E-mail address: [email protected]

Key words: types of ice cream, quality

ABSTRACT
Three types of ice cream, often used by consumers: ice cream with vanilla, cocoa and
fruit, ice cream with vanilla, cocoa and fruit and fruit ice cream, wishing -the organoleptic and
physico-chemical analysis of these ice-cream varieties with great demand among consumers.
The biotechnological assessment of the ice cream assortments under study will be based on the
fat content (milk ice cream) and the acidity (fruit ice), following certain physico-chemical changes
during storage.

INTRODUCTION
Ice cream and ice cream specialties are prepared from a special process
based on dairy products, fats, sugars, flavors, dyes, stabilizers, etc. The quality of each
ingredient is monitored from a biotechnological, microbiological, physico-chemical and
sensory (by Banu et al., 1993, 2009). The quality of fresh milk used as raw material to
obtain ice cream influences both the technological process and the quality of the
finished product (by Reithofer, 2011). Milk and dairy products have the capacity to
provide biologically active components (especially vitamins and trace elements) that
are fundamental to the promotion of human health (Scott et al., 1988, Baldi et al.,
2005). Fats can provide significant protein intake (Koxholt et al., 2001, Ruger et al.,
2002, Shaviklo et al., 2011). Ice cream is a food offering to the liking of consumers,
and because it can be obtained by mixing more raw materials, the perceived
organoleptic sensations are multiplied by the richness of nutrients found on the label
(by Tosaki et al., 2009, Dickinson, 2013).

MATERIALS AND METHODS

Determination of fat content by acid-butyric method
The principle of the method consists in separating the fat in the butyrometer by
centrifugation after the preliminary dissolution of the protein substances in sulfuric acid
in the presence of isoamyl alcohol.
Working mode Add 10 cm 3 of H2SO4 and 5 cm 3 of ice cream thoroughly
3
homogenised in the butyrometer. The frozen pipette is rinsed with 6 cm of distilled
water and added to the butyrometer then 1 ml of isoamyl alcohol is added. Remove the
butirometer with cotton wool, screw the rubber stopper and mix. After homogenization,
the butyrometer is centrifuged for 5 minutes at 1000-1200 rpm, removed from the

63
centrifuge and placed on the water bath at 65°C. Read the fat content on the
butirometer rod and the reading read multiplied by 2.2.
Determination of total acidity of frozen is done by the current titration method,
which applies to milk and all dairy products, including ice cream. The acidity of a given
volume of sample prepared for analysis is neutralized with a 0,1 n NAOH solution in
the presence of phenolphthalein as an indicator. It is expressed in Thörner grade =
acidity in 100 cm 3 of the product which is neutralized with 1 cm3 of NaOH solution 0,1n
after calculating and interpreting the results.
Determination of dry substance content consists of using a special balance
(Lacta). Introduce 5 g of paraffin and 5 g of ice cream. Heat moderately for evaporation
and shake vigorously. When the clock glass is no longer steamed, the evaporation is
complete. Keep in desiccator 10 minutes after balancing on the balance with 2 riders.
Rider. 1 (2 g) 1 div. = 0.4 g; Rider. 2 (0.5 g) 1 div. = 0.1 g x • 0.4 g + y • 0.1 g = zg
water of 5 g frozen.
Determination of sugar from ice cream
Determination of sugar is made by the iodometric method in two variants. The
aldehyde group of the reducing sugar is oxidized with iodine in alkaline medium.
Excess iodine is titrated with sodium thiosulphate solution in the presence of starch.
The titre of the sodium thiosulphate solution (T) expressed in grams of sucrose per ml
of solution is calculated according to the formula and expressed in g/cm3.

RESULTS AND DISCUSSIONS

The nutritional impact of ice cream ingredients is often transposed into their
biotech assessment to determine how they can influence consumer health.
The biotech assessment of ice frost during the hot season is of particular importance
and is based on several aspects. It primarily concerns the quality of the raw materials
that are part of the ice cream, the strict observance of the technological process and
last but not least the keeping and transportation to the beneficiary under appropriate
conditions.
In order to determine the nutritional value of the frozen types of ice cream, the
chemical analysis of the main compositional elements was performed. The data
obtained are listed in Table 1. Greater fat content is found for cream based ice cream
than for milk. (8.5% versus 3.5%). In the frozen fruit, the fats are missing. Figure 1
shows the large difference in fat content of the three types of ice cream analyzed.
Concerning the sugar content, it can be seen that both types of ice cream (milk-based
and cream-based) have a similar content in sugar (14-15%), while fruit ice has
recorded values close to twice as high compared to other assortments analyzed (27%
versus 14%).
Table 1
Composition analysis of the main types of ice cream

Types of ice cream Fat Sugar Dry substance Acidity

(%) (%) (%) (ºT)
Of milk:
- with vanilla, cocoa 3,5 14 27 20
- with fruit 2,8 15 28 46
By the cream:
- with vanilla, cocoa 8,5 14 32 21
- with fruit 7,0 15 31 47
Of fruit:
- with fruit flavor - 27 27 68
64
 9 8,5
8 Milk ice cream with
7 vanilla and cocoa
7
Frozen milk ice cream
6
5 Ice cream and vanilla
% 3,5 ice cream with cocoa
4 2,8 Ice cream with fruit
3
2 Frozen ice cream
(strawberry)
1
0
0
fat content

Figure 1 Determination of the fat content of the studied ice cream

And acidity, expressed in degrees Thorner, recorded similar values for milk
and cream ice cream (20-21ºT). The opposite is the ice cream with fruit, whose acidity
was 68 degrees tall. And within the same assortment (eg ice cream), the fruit has
higher acidity (46ºT to 20ºT) - figure 2.

Frozen ice cream

68 (strawberry)
47 Ice cream with fruit
acidity

21 Ice cream and vanilla

and cocoa
46
Frozen milk ice cream
20
Milk ice cream with
vanilla and cocoa

0 20 40 60 80
gr. T
.
Figure 2 Determination of acidity in the ice cream assortments taken into study

The highest content in dry matter is recorded in cream with three forms
(vanilla, 32% cocoa and 31% fruit). The lowest value in dry matter was milk vanilla with
vanilla and cocoa and ice cream (27%).
Fats and sugar, together with proteins, represent the basics that give the
nutritional value of the types of ice cream studied.
The main biotechnological aspects in evaluating a range of ice cream are
mainly transposed into ice cream defects. Ice cream presents a number of defects, the
most common being organoleptic: aroma, texture and color.
Ice cream like any food is assessed biotechologically by Stas, following the
admissibility conditions (Table 2).
The organoleptic evaluation of the ice cream samples taken in the study did
not reveal any major defects. However, it can be mentioned, from the category of flavor
65
defects, the slightly sour, poorly perceptible taste of fruit milk ice cream. An explanation
would be the use of strawberries with an average degree of freshness.

Table 2
Organoleptic characteristics for ice cream

Characteristics Admissibility conditions

Clear, characteristic, for ice cream of the given type and
the raw material used in its manufacture, with no foreign
flavors and smells; taste sweet or sweet-sour.To use the
food ingredients to taste and smell - must match the taste
and flavor of the ingredient introduced. When using
flavorings - it must match the flavor and aroma of the flavor
Taste and aroma enhancer. There is insufficient taste or scent or excessive
odor of the flavoring used. When using glaze coating and
food ingredients for decoration - it must match the given
type of ice cream in combination with the taste, aroma and
smell of the coatings used and the decoration ingredients.
No smell and taste of fodder, seasons, rînced, fish, burn,
metal, mineral, salty, alien. It does not allow insufficient or
excessively sweet taste.
Consistency Dense enough for tanned ice cream. Dish for food ice. Soft,
creamy for soft ice cream.
Homogeneous, without perceptible organoleptic
agglomerations of fat and stabilizer (stabilizer-emulsifier).
When using the food ingredients with the appearance of the
pieces - with the presence of their particles. No perceptible
organoleptic ice crystals - for soft and tempered ice cream.
Appearance in It is not allowed the structure and consistency of snow, ice,
section flakes, flour, sand, rock, hard fusible. The tiled ice cream
structure with a fat mass fraction of not more than 5% and a
dry mass fraction of dry matter of not more than 30% and
ice cream based on sugar syrup are allowed. For glaze ice
cream the color of the coating - characteristic of the given
type of glaze or chocolate.
For tanned ice cream with a good shape, without
deformations. Admitted (up to 10 mm) small spills of ice
cream and / or glaze over waffles or biscuits. Cracks and
Exterior other insignificant mechanical damage (not more than 10
appearance mm) of glaze or waffles, crackers, including edges of waffle
products are allowed. In glazed ice cream the glaze must
cover the ice cream tightly. It is not allowed to shrink and
detach ice cream from waffle and biscuit products.

The beginning of the defect is also due to the high acidity found in this ice
cream assortment. The taste and smell of the rancid was detected in the case of ice
cream with cocoa cream, and was largely due to the use of inappropriate raw materials
or the non-observance of the stages of the technological process of production (the
basic mixture was maintained too long before to be frozen in the presence of air,
resulting in the phenomenon of oxidation of the fat in the composition).

66
 The texture defect such as the sandy texture has also been encountered in the
fruit frost, the factors that lead to the formation of a non-uniform mixture are: the poor
quality of the raw materials used, the composition of the mixture, the processing
method, the freezing and quenching process. Thick texture met with ice cream and
vanilla ice cream, persisting for a longer time than fat.
Color defects were encountered in fruit ice (the intense pink color specific to
strawberries was not homogeneous in the mass of the product) and in milk cocoa ice
cream, where the intense cream color was also uneven in the mass of the product.

CONCLUSIONS
Following the determinations, there is a higher fat content in whipped cream
based on cream than milk, fruit ice cream, and fats lacking.
Fruit ice cream sugar content has almost doubled compared to the other assortments
analyzed. Acidity, expressed in degrees Thorner, recorded similar values for milk and
cream ice cream and higher for fruit ice, both as a single assortment and in other
assortments.
The highest content in dry matter is recorded in whipped cream with the three
forms (vanilla, cocoa and fruit), a lower dry amount found in vanilla ice cream and
cocoa and fruit ice cream.
The main biotechnological aspects in evaluating a range of ice cream are
mainly transposed into ice cream defects.
Some of the ice cream varieties analyzed showed a number of defects, the
most common being organoleptic: flavor, texture and color. So:
 Fruit milk ice cream: flavor defects (slightly sour, poorly perceived taste);
 Coconut ice cream: taste and smell easily rancid;
 Fruit ice cream: sandy texture, color defects.

REFERENCES
Constantin Banu, Berzea Marilena, Camelia Vizireanu, Toader Roxana, 1993,
Totul despre îngheţată, Editura Tehnică Bucureşti
Constantin Banu, 2009 Tratat de industrie alimentară, Tehnologii alimentare,
Editura ASAB, București
Dickinson, E. 2013, Stabilising emulsion-based colloidal structures with mixed
food ingredients. J. Sci. Food Agric., 93: 710–721. doi:10.1002/jsfa.6013
Baldi, A., 2005 Vitamin E în dairy cows. Livestock Production Science, 98 (1), 117-
122
Căpruciu Ramona, 2013, Biotehnologii moderne în industria alimentară, Editura
Sitech, Craiova
Gholam Reza Shaviklo, Gudjon Thorkelsson, Kolburn Sveindothir, Fereidon
Rafipour, 2011, Chemical properties and sensory quality of ice cream fortified with fish
protein, Journal of the Science of Food and Agriculture, Volume 91, Issue 7, 91:
1199–1204. doi:10.1002/jsfa.4299
Koxholt N M R, Eisenmann B, Hinrichs J, 2001, Effect of the fat globule sizes, on
meltdown of ice cream. Journal of Dairy Science 84 31–37
Elisabeth-Mayer Reithofer, 2011, Produse lactate- tehnologii și rețete de obținere
Editura MAST
Ruger P R, Baer R J, Kasperson K M, 2002, Effect of double homogenization and
whey protein concentrate on texture of ice cream. Journal of Dairy Science 85 1684–
1692
67
 Scott, K. J. and Bishop, D. R., 1988, Nutrient content of milk and milk products:
Vitamins of the B complex and vitamin C in retail creams, ice creams and milk shakes.
J. Sci. Food Agric., 43: 193–199. doi:10.1002/jsfa.2740430210
Tosaki, M., Kitamura, Y., Satake, T., Tsurutani, T. 2009, Effects of
homogenisation conditions on the physical properties of high-fat ice cream.
International Journal of Dairy Technology, 62: 577–583. doi:10.1111/j.1471-
0307.2009.00525.x

68
 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
 Food produce process ing
UNIVERS ITY OF CRAIO VA
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

ASSESSMENT OF THE TECHNOLOGICAL QUALITIES OF THE SOME

INTRASPECIFIC HYBRID ELITES FOR THE TABLE GRAPES COMPARED
WITH THE PARENTAL FORMS BY THE METHOD OF THE DISTRIBUTION
ON THE FREQUENCY CLASSES
1 1
Ciobotea Cristina-Magdalena , Popa Camelia , Radomir Ana-Maria
1*
N. I. R. D. B. H. Stefanesti-Arges, Sreet Bucharest - Pitesti, No.37,Stefanesti, [email protected]

Key words: technological descriptors, distribution on the frequency classes, hybrid elites,
parental formes

ABSTRACT
The present paper reveals the features and technological characteristics of 21 hybrid
elites resulting from the hybridization of four soups for table grapes in three distinct combinations
(Muscat Iantarni x Canner, Victoria x Black Pearl and Victoria x Victoria) selected from the
comparatively field, and also highlights the combined value of the varieties used in the
amelioration process by means of distribution by frequency classes.
In order to highlight the technological value of the genotypes obtained from the three
combinations mentioned above as well as the combination potential of the varieties involved in
the breeding process, the frequency distribution of the average values obtained was used. The
multitude of the data obtained was statistically and mathematically processed, based on the
descriptive statistics, with general clues on the way of manifestation of the hybrid vines
genotypes in the interaction with the biotic and abiotic factors.

INTRODUCTION
Based on data obtained, by statistical and mathematical processing, by the
descriptive statistics, general conclusions have been formulated concerning the
manner of manifestation of the hybrid elites grape-vine in interaction with the biotic and
abiotic factors having regard to natural variability of all processes and phenomena
involved.
This presents two aspects: - Representation of the entire distribution, by:
numerical (frequency analysis) and graphics (bar, histogram, box plot, stem and leaf);
central tendency indicators: mode, average, median; dispersion indicators: amplitude,
standard deviation; the indicators of the distribution form; index of asymmetry
(skewness) and index of vaulting (kurtosis).
Skewness (skewness) and Kurtosis (kurtosis) în SPSS - Skewness and
kurtosis values are normal and equal to 0. In the case of a random sample, extracted
from a normal population, it is unlikely to obtain the indices of symmetry and vaulting
equal to 0. Must be known if the sample of the analyzed values are derived from a
population asymmetric or abnormal vaulte.
The utility of the normality test of the distribution relates to the decision to use
parametric tests in the case of the quantitative variables, which are subjected to this
condition. Both the K-S test and Shapiro-Wilk are sensitive to the asymmetry and
69
vaulting. Therefore, when it is used the t test or analysis of variance, vaulting is less
important than the asymmetry. For this reason, if the test of normality is significant, it is
advisable to check whether this is due to the asymmetry, vaulting or both. For both
tests (Kolmogorv-Smirnov and Shapiro-Wilk), is only important the value of p (Si G.).
this being understood inversely than the classical interpretations of the p.

MATERIAL AND METHODS

In order to highlight the technological value of the hybrid elites resulting from
three combinations (Muscat Iantarnii x Canner, Victoria x Black Pearl, Victoria x
Victoria) and the combinational value of the varieties involved in the breeding process,
the frequency distribution of the obtained mean values was used. Were introduced into
the histograms all the values of the three types of hybrid descendants with the parental
varieties (26 genotypes), for characters analyzed. For more meaningful representation
of the data, it used the class frequency distribution of the average values of parameters
determined for elite hybrid and parents.
From here results that the result interpretation of the normality test is
determined by the simultaneous correlating of the p-value with the sample size. The
Gauss distribution for the normal values, in the form of a bell, is represented by two
values: the average value and standard deviation.

RESULTS AND DISCUTIONS

A symmetrical distribution and only slightly platykurtic compared to the average
was recorded in the case of yhe average weight of a grape. The coefficient of variation
of the values around the mean was average (25,9%), the maximum oscillation limits
being between 150 minima and 565 maxims.
With an average grape weight of 565 and 560 g, the selected elites A1 and
A19 are at 4 frequency classes in addition to the average. Low values below 250 g
show only the Muscat Iantarnîi variety of the maternal genitor. All the elites had grapes
heavier than 250 g (figure 1).

Greutatea medie a unui strugure (g)

4
Frequency

2 4 4 4

3 3

1 2

1 Mean =395,88
Std. Dev. =102,682
0 N =26
100 150 200 250 300 350 400 450 500 550 600

Greutatea medie a unui strugure (g)

Figure 1 - Distribution on the frequency classes of average values of the average
weight of a grape, to the hybrid elites

Also an empirical distribution with right asymmetry and weak leptokurtic is

indicated in the case of the average weight of 100 berries. The maximum oscillation of
70
the indicator was 1165 g by weight of 100 berries (between 234 and 1400). The
average was 524,96 and the higher coefficient of variation (53,3%) the sample being
more heterogeneous. Asymmetry is the right, which shows that dominates the elites
with the value of the weight grape less than average. The vaulting coefficient had a
value of 3,901, which shows the occurrence of excess values only near the mean and
far from it, with few representations in the high-value classes (figure 2).

Figure 2 - Distribution on the frequency classes of average values of the average

weight of the 100 berries, for F1 elites

Selected elites were distributed more around the average, but there were 2
genotypes with double values from the mean: A16 (1400 g) and A19 (1100 g). Lower
values recorded the hybrid descendents resulting from Muscat Iantarini x Canner
combined, character influenced by the berries of the maternal variety.
The content of the must in sugars ranged between 110 and 210, the maximum
variation amplitude was 100 g / l, and the deviation of the values around the average of
160,15. The coefficient of the variation was only 15%. The deviation from normal was
reduced. The elites were in a large number around of the average of 160,15 (15 elites),
with the values of the sugars from the must ranging from 140-180 g/l. The lowest value
for this indicator was the Hi10 elite (110 g / l), and the highest content, the A31 elite
(210 g/l) (figure 3).
The histogram of the must acidity for the hybrid descendents has the following
characteristics: the mean value was 3,031 g/l of H 2SO4 and the coefficient of variation
of 19,6%.
The excess of the sample was close to the normal distribution. There was a
slight right asymmetry, which means that values below the mean prevailed. With acidity
below 2,2 g / l of H 2SO4, the elites from the BP group (BP2, BP9, BP11) and a group
A (A17) elite predominated, and with the highest level of acidity, the group Hi (Hi3 and
Hi11) (figure 4).

71
Figure 3 - Distribution on the frequency classes of average values of the must content
in sugar, to the hybrid genotypes

Figure 4 - Distribution on the frequency classes of average value of the must acidity,
for F1 hybrids

CONCLUSIONS
Conclusions on the study of the technological characters of elites resulted by
Muscat Iantarnii x Canner hybrid combination (Hi) showed the following points:
 all elites had larger berries than the hybridized variety (Muscat Iantarnii)
and two elites (Hi17 and Hi19) passed the Canner variety to this descriptor;
 the sugars content to hybrid descendents from this combination (Hi) ranged
from 110 g/l for the Hi13 elite to 180 g/l at the Hi17 elite; the Hi19 elite accumulated
160 g/l, being overtaken by three elites, including Hi9 and Hi11 (175-170 g/l);

72
  three elites (Hi11, Hi17 and Hi19) inherited from the maternal variety the
muscat taste,and an elite (Hi13) acquired from the paternal genitor the specific taste,
and another elite presented the frank taste;
 all elites had the greenish-yellowish berry.
Conclusions on the study of the technological characters of elites resulted
by Victoria x Black Pearl (BP) hybrid combination showed the following points:
 because the varieties with large grapes participated in the hybridization
process, it is noted that the descendants presented large and very large bunch;
 the Black Pearl paternal variety was exceeded, in regards the weight of
bunch, by five elites (exception only makes BP2 elite with 310) and Victoria variety with
weight 470 g was equalized by the elite BP11 (470 g); the rest of the descendants
presented have values above 400 g, respectively higher than the average of all
analyzed genotypes;
 the Black Pearl variety was overtaken by the weight of 100 berries by the all
the elites and the Victoria variety rated and considered standard for the large berry,
was surpassed by the BP 11 elite, with 20 g (650 g, respectively 670 g); four hybrid
elites can be ranked in the large berry class because they have values above 500 g,
well above the average of 493 g; with regard to the sugar content, BP8 elite is
particularly noteworthy, with 206 g/l, the value rarely encountered in table grape
varieties, and especially in hybrid elites;
 the acidity of the must showed close values between them, three elites
placed above the average and cross-breeds varieties: BP2, BP7, BP8, with
concentrations between 3,3-3,8 g/l H 2SO4;
 it is noted that the BP 2 and BP 7 descendants have a discrete flavor, the
rest of the elites beeing whitout flavor;
Conclusions on the study of the technological characters of elites resulted
by Victoria x Victoria hybrid combination (A) showed the following points:
 Victoria variety was surpassed in the average weight of 100 berries by the
six of the 10 selected hybrid elites (A1, A5, A16, A17, A19, A27);
 sugars content higher than Victoria (141 g/l),we meet at the elites: A1, A5,
A9, A6, A16, A19, A25, A31;
 it was found that there is a significant positive and linear correlation
between the weight of 100 berries (g) and the weight of a grape (g) to the all the
studied elites;
 it is interesting and surprising at the same time that the elites A1, A9, A25
have the muscat flavor, elites A5, A6, A16, A17 possess a specific flavor;
 muscat flavor is intense at A1 and A9 and discrete at A6 and A 25.

REFERENCES
Cichi Daniela, Popa Camelia, Necula Cezarina, 2010, Ghid ampelografic al
soiurilor de struguri pentru masă, Ed. Universitaria, Craiova, 50-54 ;
Coelho de Souza Leão.,Cosme Damião Cruz, Sérgio Yoshimitsu Motoike, 2011-
Genetic diversity of table grape based on morphoagronomic traits, Scienta Agricola
(Piracicaba, Braz.) vol.68 no.1 Piracicaba;
Damian Doina, Calistru Gh., Savin Gh., Abruda P., 1997, Potenţialul agrobiologic
al unor soiuri apirene studiate în ecosistemul viticol Copou-Iaşi, Cercet. agr., în
Moldova, Vol.3 (109), 47-53;
Gribaudo I., Grando M.S., 2005, Caratterizzatione varietale e miglioramento
genetico della vite. In: Marenghi M., Manuale di viticoltura, Impianto, gestione, e difesa
del vigneto, Edagricole Bologna, 1-15;

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 Meirong Y., Yingzhu Z., C., Shengchen L., 1995, Breeding for early ripening table
grape varieties, ISHS Acta Horticulturae 403, International Symposium on Cultivar
Improvement of Horticultural Crops;
Onea (Ciobotea) Cristina-Magdalena, 2014, Studiul caracterelor ampelografice,
agrobiologice şi tehnologice ale unor hibrizi intraspecifici noi pentru struguri de masă
obţinuţi la Ștefăneşi - Argeş, Teză de doctorat, Universitatea din Craiova,
Oprea St., Pamfil D., Olaru B., 1989, Aplicaţii genetice în ameliorarea viţei de vie,
Cercet. Genet. Veget. Anim., I, 247-263;
Pearl A., Sahar, N., Eliassi, R., Baron, I., Spietel-Roy, P., Bazak, H. 2003 -
Breeding of new seedless table grapes in Israel conventional and biotechnological
approach. Acta Hort. (ISHS) 603:185-187;
Shapiro S.S., Wilk M.B., Chen M.J., 1968, A comparative study of various tests for
normality. Journal of the American Statistical Association, 63;

74
 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
 Food produce process ing
UNIVERSITY OF CRAIO VA
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

STUDY ON THE BEHAVIOR OF CAPSICUM

GRAFTED ON DIFFERENT ROOTSTOCKS
IN SANDY SOIL CONDITIONS
1 1 2
Ciuciuc Elena , Croitoru Mihaela , Vânătoru C.
1
Research-Development Center for Field Crops on Sandys Soils Dabuleni
2
Development Research Center for Vegetables Buzău
Corresponding author: [email protected]

Key words: capsicum, grafting, rootstock

ABSTRACT
Decreasing or eliminating the attack of pests and soil pathogens can be achieved by
introduction grafting crop technology. For this purpose Was used as a graft the Ișalniţa 85 V
variety grated and as rootstocks was used 6 lines obtained at SCDL Buzău (L 12, L 60, L63,
L66, L70E și L70P). Determinations were made on the main physiological processes, the height
increases of the capsicum plant, the plant health status, the number of fruit / plant, the average
weight of a fruit, the total and dynamic production and the biochemical content of the fruit. At the
first harvesting by grafting on the L66 rootstock, the highest yields were achieved, ie 13.8 t / ha,
with 1 t / ha more than plants without grafting, and the smallest production was made on grafted
plants on L12 . Total production was 40.0 t / ha for plants without grafting and between 32.9-43.8
t / ha for grafted plants. Were noted by the production of the plants grafted on L 66 with 43.8 t /
ha and the grafted on L 70 E by 43.2 t / ha. The plants grafted on the L66 rootstock were
distinguished by a well developed root system, with thick, strong roots associated with the lack of
nematodes, and contributed to significant production gains.

INTRODUCTION
The newly introduced pepper grafting has emerged as a necessity for
controlling pathogens and nematodes in the soil and for increasing the tolerance or
resistance to stress factors. In This has scope was studied numerous of rootstock for
compatibility of the capsicum culture.
Attia M.F. et al., 2003 demonstrated that solanaceae rootstocks were accepted
as compatible for both sweet pepper culture and pepper culture. The effect of pepper
grafting on Fusarium infested soil was conclusive.
The effect of pepper grafting on Fusarium infested soil was conclusive. Maria
Maribel Rodriquez and Paul W. Bosland, 2010, studied the compatibility of tomatoes
as rootstocks for Capsicum annuum L. and concluded that grafting peppers on
tomatoes provides a mechanism for controlling soil and nematode diseases that
negatively affect pepper and that Is still worth investigating. The older paprika plants
had a good development, connection to the graft site compared to the young plants.
The authors also conclude that rootstock root can be crucial for leaf water content,
especially under stress conditions. The fact that grafting reduces stress was also
reported by other authors (Lee J.M. et al., 1998, Rivero R.M., 2003, Shibuya T. et al.,

75
2007, Carmen Martinez-Ballesta, 2010). The authors believe that the physiological and
biochemical aspects of the graft-rootstock interaction should be studied.
The nutritional qualities of grafted pepper plants such as total dry substance,
soluble dry matter, titratable acidity are comparable of the plants without grafting,
while of grafted plants increased vigor and resistance to soil and nematode diseases
(Colla G. et al. , 2008; Davis AR, 2008a). For the management of nematodes and the
replacement of methyl bromide by grafting the pepper plants choose and other
authors (Lee J.M. et al., 1998, Attia M. F. et al., 2003, Morra L and Bilotto M., 2006,
King S.R., 2008).

MATERIAL AND METHOD

At the Research-Development Center for Field Crops on Sandy Soils Dabuleni
was studie the behavior of the Isalnita 85 V grape variety was grafted on various
rootstocks obtained at SCDL Buzău: L 12, L 60, L 63, L 66, L 70 E, L 70 P, compared
to the plants without grafting.
Was determine of the rootstock influence on the health status of the capsicum
plants (identification of pathogens of soil and nematodes), determinations of
productivity (number of fruit / plant, average weight of fruits) A fruit, total and total
sweet pepper production) and the determination of the biochemical content of the fruit.
The results were calculated and interpreted statistically.
Seedlings were obtained in the double-shielded solar hood. The grafting
method used was that of "sticking with a cotyledon”.
Fruit harvesting was done at the consumption maturity, staggered.

RESULTS AND DISCUSSIONS

The smallest elevations in were made in plants grafted on L 70 E and L 70 P,
and the largest waistline was recorded at Isalniţa 85 V grafted on L60 (Table 1). The
number of fruit/plant was influenced by the rootstock used being of 14.2 fruit/plant on
the without grafting plants and ranging from 11.7-15.6 fruit/plant to the grafted plants.
The largest number of fruits was recorded in plants grafted on L 66 and L 70 P. The
average weight of a fruit oscillated within very narrow limits and ranged from 50-53
g/fruit.

Table 1
Biometric determinations made

The average
Rootstock Plant height Number of fruits / weight of a fruit
(cm) plant (g/fruit)

Without grafting 35.6 14.2 50

L12 37.4 14.0 51
L60 42.2 14.4 53
L63 35.8 14.8 53
L66 33.4 15.4 51
L70 E 33.0 15.6 50
L70 P 30.6 11.7 51

Five crops were taken as the fruits matured. The first harvest took place on
July 22, and the productions made at that date were greatly influenced by the rootstock
used (table 2).
76
 The Isalnita 85 V cultivar without grafting achieved on July 22 a production of
12.8 t/ha, which was exceeded by the production obtained from the L66 grafted plants
at 13.8 t/ha and to those grafted on L60 that achieved 12.2 t/ha. The lowest yields
were obtained at this time on L12 grafted plants (5.5 t / ha), L70P (7.2 t / ha) and L63
(9.4 t / ha).
Table 2
The dynamics of the production of capsicum according to the rootstock used
Production of capsicum obtained on (t / ha):
Rootstock 22.07 14.08 26.08 9.09 23.09

Without grafting 12.8 11.6 5.0 5.0 5.6

L12 5.5 7.2 10.0 11.1 6.1
L60 12.2 4.4 7.8 5.0 13.3
L63 9.4 5.0 6.7 8.9 11.1
L66 13.8 6.7 7.2 10.0 6.1
L70 E 11.1 6.1 9.4 9.4 7.2
L70 P 7.2 3.4 5.6 11.1 5.6
At the second harvest, on August 14, the production of without grafting plants
was maintained at a high level, of 11.6 t / ha, while at grafted plants were low, ranging
from 3 , 4-7.2 t / ha. At the following harvests, the yields of without grafing plants
decreased and were maintained at high levels in grafted plants, the rootstock being
resistant to thermal stress conditions.
Total capsicum production was influenced by the rootstock used (table 3). The
Isalnita 85 V variety produced a 40 t / ha production in without grafting crops,
compared to which, by grafting on the L12 rootstock, a very close production was
achieved, the difference being only -0.1 t / ha.

Table 3
Influence of rootstock on the production of capsicum
Rootstock Production achieved Difference Signifficanc
t/ha % (t/ha) e
Without grafting 40.0 100 Mt. Mt.
L12 39.9 100 -0.1
L60 42.7 107 +2.7
L63 41.4 104 +1.4
L66 43.8 110 +3.8
L70 E 43.2 108 +3.2
L70 P 32.9 82 -7.1 0
DL 5% = 6.46 t/ha DL 1% = 8.79 t/ha DL 0,1% = 11.77 t/ha

Using the L60 rootstock a production of 42.7 t / ha and a production increase

of 2.7 t / ha was achieved, and by grafting on the L63 rootstock there was an increase
of 1.4 t / ha. The production of the without grafting variety was also exceeded by that
obtained with plants grafted on the L 70 E and L 66 rootstocks, the production yields of
which were 3.2 t / ha using the L70 E rootstock and 3.8 t / ha at the L66 rootstock.
The smallest production obtained by grafting the Isalniţa 85 V variety was obtained by
grafting on the L70 P rootstock, this being 32.9 t / ha, the production difference being
7.1 t / ha.

77
 After the last harvest, the root system was unveiled and the presence /
absence of nematodes was determined and the degree of root development based on
the rootstock used (table 4).
Table 4
Highlighting the nematode attack on grafted plants on the studied rootstocks

Rootstock Presence / Remarks

absence of
nematodes
Without Preence Very few nematodes, well developed root system,
grafting thick roots
L12 Absence Rich radicular system, thin roots
L60 Absence Radical lax system
L63 Absence Weak root system developed
L66 Absence Root system developed, thick roots, strong
L70 E Presence Nematodes in very small number, lax root system,
thick roots, few
L70 P Presence Much nematodes, root system developed

The rootstock L12 showed a root system rich in roots, but very thin, a poor
presence of nematodes, the production obtained by it being very close to the without
grafting witness. The rootstock L60 and L63, although presented with a radicular
system with few roots, the lack of nematodes positively influenced the produced
productions.
Among the studied rootstocks, a well-developed root system with thick, strong
roots associated with the lack of nematodes has contributed to the achievement of
significant L66 production increases.
The rootstock used has influenced not only quantitative but also qualitative
production (table 5).
Table 5
The influence of the rootstock on the quality of the capsicum fruit

Titratable
Total Solubil
acidity Carbohy- Vitamin C
Water dry dry
Rootstock g citric drates (mg/100
(%) substance substance
acid in (%) g f.s)
(%) (%)
100 g f.s.
Without 93.96 6.04 4.2 0.19 3.50 21.12
grafting
L12 93.68 6.32 4.6 0.36 3.80 29.92
L60 93.69 6.31 4.4 0.19 3.70 40.48
L63 94.67 5.33 4.4 0.36 3.68 36.08
L66 93.17 6.83 4.6 0.18 3.85 38.72
L70E 94.38 5.62 4.0 0.19 3.35 36.96
L70P 93.54 6.46 4.6 0.23 3.82 36.08

The following determinations were carried out in the laboratory: water content
and total dry substance (gravimetric method), soluble dry matter (refractometric
method), total sugars (Fehling Soxhlet method), titratable acidity (titrimetric method),
vitamin C (iodometric method).

78
 Total dry matter content ranged between 5.33% for Isalniţa 85 V grafted on L
63 and 6.83% grafted on L70P. Compared to the without grafting control in most
grafted variants, the amount of total dry substance showed higher values. The soluble
dry substance content, in all grafted variants, showed values comparable to those of
the without grafting control and was between 4-4.6%.
In peppers, the carbohydrate content is in the form of glucose, fructose and
sucrose, the amount of glucose and fructose being higher than that of sucrose.
Grafting of plants has favorably influenced the accumulation of sugars in peppers, and
the best results were obtained at the fruit of the Isalnita 85 V cultivar grafted on L66
(3.85%).
All pepper varieties contain vitamin C in large quantities, but green capsicum
has twice as much vitamin C as citrus, and red pepper contains 330% more vitamin C
than orange. In 100 grams of pepper are 125 mg of vitamin C, that is, 100% of the
recommended daily dose.
In all grafted variants, higher values of vitamin C content were obtained
compared to the without grafting control.
If we analyze the influence of the rootstock on the biochemical composition of
the fruit, the best results were obtained in the variant where the plants were grafted on
the L66.

CONCLUSIONS
At the first harvesting by grafting on the L66 rootstock, the Işalniţa 85 V variety
produced the highest yields of 13.8 t / ha and the production was low when using the
L12 rootstock.
The biggest productions were obtained by grafting on the L66 rootstock. These
were 43.8 t / ha providing a production increase of 3.8 t / ha compared to without
grafting. All biochemical components of the fruits showed higher values in plants
grafted on L 66. The high yields produced by grafting on the L66 rootstock were due to
the fact that it was distinguished by a well-developed root system with thick, strong
roots associated with the lack of nematodes and contributed to significant production
increases.

REFERENCES
Attia M.F., Arafa A.M., Moustafa M.A., Mona A. Mohamed, 2003 – Pepper
Grafting, a Method of Controlling Soilborne Diseases and Enhancement of Fruit Yield:
1. Improvement of Pepper Resistance to Fusarium wilt. Egypt. J. Phytopathol. Vol. 31,
No. 1-2, pp 151-165.
Carmen Martiney-Ballesta, Carlos alcaraz-Lopez, Beatriz Muries, cesar Mota-
Cadenas, Micaela Carvajal, 2010 – Physiological aspects of roostocks-scion
interaction. Scientica Horticultural 127: 112-118.
Colla G., Rouphael J., Cardarelli M., Temperini O., Rea E., Salerno A., Pierandrei
F., 2008 – Influence of grafting on zield and fruit quality of pepper (Capsicum annuum
L.) grown under greenhouse conditions. ISHS Acta Horticulturae 782: IV International
Symposium on Seed, Transplant and Stand Establisment of Horticultural Crops;
Translating Seed an Seedling Phisiology into Technology.
Davis A.R., Perkins-Veazie p., Hessell R., Levi A., King S.R., Zhang X.P., 2008a –
Grafting effects on vegetable quality. HortScience 43, 1670-1672.
King S.R.,Davis A.R., Liu W.G. and Levi A., 2008 – Grafting for disease
resistance. HortScience 42: Pg. 1673-1676.
Lee J. M., Bang H. J. et all.,1998 – Grafting of vegetables. Journal of the
Japanese Siciety for Horticultural Science 67 (6): 1098-1104.
79
 Maria Maribel Rodriquez and Paul W. Bosland, 2010 – Grafting Capsicum to
Tomato Roostocks. The journal of Joung Investigators. Volume 20, Issue 2.
Morra l., Bilotto M., 2006 – Evaluation of new roostocks for resistance to soilborne
pathogens and productive behaviour of pepper (Capsicum annuum L.). J. of
Horticultural Science and Biotech. 81: 518-524.
Rivero R. M., Ruiz J.M. et all., 2003 – Role oh Grafting in Horticultural Plants
Under Stress Conditions. Food, Agriculture & Environment 1(1): 70-74.
Shybuya T., Nakashima H., Shimizu-Maruo K., Kawara T., 2007 – Improvement of
graft development in tomato and eggplant cuttings by supplying warmed water to graft
union during low-air-temperature storage. Hortic. Sci. 76, 217-223.

80
 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
 Food produce process ing
UNIVERSITY OF CRAIO VA
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

DEVELOPMENT AND EVALUATION OF A REVERSE-PHASE HPLC

METHOD FOR THE ANALYSIS OF CAROTENOIDS IN EGG YOLK

Corbu Alexandru Radu1, Violeta Nour2*

1
"Dunarea de Jos" University of Galati, Romania
2
University of Craiova, Romania
Corresponding author e-mail: [email protected]

Key words: egg yolk, carotenoids, chromatography, method validation

ABSTRACT
Carotenoids from the egg yolk are important pigments influencing the consumer
acceptance and essential components associated with health benefits either as antioxidants and
immunomodulants or as precursor of vitamin A. Recently ther ehas been considerable interest in
enhancing the levels of yolk carotenoids by including carotenoids or carotenoid rich forage
products in laying hen diets. In order to learn the variety and amount of carotenoids in regular
and modified eggs, diiferent chemical analytical methods have been developed. In the current
study, a reversed-phase high performance liquid chromatography (RP-HPLC) method with
gradient elution and diode array detection (DAD) was developed and validated for the
simultaneous determination of seven carotenoids (lutein, zeaxanthin, canthaxanthin,
astaxanthin, lycopene, -carotene and trans-β-apo-8'-carotenal) in egg yolk.

INTRODUCTION
Hen eggs are a very popular food of animal origin, being eaten by many
people all over the world. They are served in various ways and they are widely used in
cookery for leavening, thickening, binding, and emulsifying (Surai & Sparks, 2001). It is
well-known their high nutritional value, due primarily to their great content of highly
digestible and complete nutritional proteins containing essential amino acids in a profile
similar to the ideal balance of amino acids needed by human (Regal et al., 2014). As a
result, the protein of the egg is used as a standard for measuring the quality of other
food proteins (Sparks, 2006).
Eggs contain high amounts of lipids (aprox. 11%) consisting of triacylglycerols,
phospholipids and cholesterol, more than 99% being located in the yolk (Spark, 2005).
Eggs are an important and balanced source of essential fatty acids as well as of some
minerals and vitamins. They contain significant amount of phosphorus, iodine,
selenium and iron as well as vitamins E, A, B12, B2 and folate (Surai & Sparks, 2001).
Egg yolks serve as an important dietary source of highly bioavailable
carotenoids that are essential components with positive health effects either as
antioxidants or as precursor of vitamin A (Chung et al., 2004; Islam & Schweigert,
2006). Based on their structure, carotenoids are divided into two groups: carotenes
and xanthophylls. Carotenes are hydrocarbon containing carotenoids (e.g., β-carotene
and lycopene) whereas xanthophylls are oxygenated derivatives (e.g., lutein and
astaxanthin) (Rivera et al., 2011; Gayosso García Sancho et al., 2011). Nutritionists

81
increasingly recognise the ability of carotenoids to scavenge free radicals and to affect
innate, humoral and cell-mediated immune response (Sparks, 2006). The consumption
of carotenoids has been associated with a number of health benefits, such as cancer
chemoprotection, prevention of heart and vascular disease, and prevention of other
chronic and degenerative diseases (e.g., Alzheimer„s Disease) (Regal et al., 2014).
Lutein, zeaxanthin,, canthaxanthin, β-cryptoxanthin, β-apocarotenoic ester and
β-carotene are the main carotenoids that have been identified in the yolk of hens‟ egg.
Lutein and zeaxanthin are reported to be associated with a reduced incidence of age-
related macular degeneration (Goodrow et al., 2006), a major cause of deterioration of
sight in elderly people (Alves-Rodrigues and Shao 2004; Stahl, 2005).
Carotenoids are also important pigments in eggs as the colour of the egg‟s
yolk is considered to be an important factor in determining the acceptability for the
consumer (Karadas et al., 2006). Some xanthophylls, such as lutein, cause a yellow
color while canthaxanthin cause a red color in yolks. The intensity of the yolk color and
the shade from yellow to red are dependent mainly on the concentration of carotenoids
in the diet (Knoblich et al., 2005).
Recently there has been considerable interest in improving the nutritional
quality of eggs by enhancing the levels of yolk carotenoids so as to provide the
consumer with a healthier and more balanced diet. Modified or so-called enriched eggs
have been obtained by including carotenoids or carotenoid rich forage products in
laying hen diets. Therefore, it is imperative to learn the variety and amount of
carotenoids in regular and modified eggs (Surai & Sparks, 2001; Singh et al., 2012).
Spectrophotometry (AOAC, 1996; Islam & Schweigert, 2015) and high
performance liquid chromatography (Barba et al., 2006) are the analytical methods
available for analysis of egg yolk carotenoids. The determination of individual
carotenoids such as lutein, zeaxanthin, canthaxanthin, β-carotene and β-apocarotenoic
ester in egg yolk has been routinely performed by reversed-phase HPLC (Knoblich et
al., 2005; Karadas et al., 2006; Hammershøj et al., 2010). HPLC analysis of
carotenoids is usually done with C18 or C30 RP-columns, operated with isocratic or
gradient elution using UV–vis or photodiode array or MS detection. A wide variety of
mixtures of different organic solvents were employed as mobile phases while, in order
to improve carotenoid separation, heating of the column was sometimes used. For the
extraction of carotenoids, AOAC (1993) recommends ethanol/tetrahydrofuran (THF)
(50:50 v/v) while other authors use ethyl acetate (100%) or different mixtures of
ethanol/hexane, acetone/ethanol/ hexane, ethyl acetate/hexane or acetone/hexane
(Barba et al., 2006).
This work aimed at developing and evaluating a suitable, reliable, rapid and
simple HPLC method employing a C18 column and UV–Vis photodiode array detection
(DAD) for simultaneous determination of carotenoids in egg yolk.

MATERIALS AND METHOD

Reagents and standards
Standards of lutein, zeaxanthin, canthaxanthin, astaxanthin, lycopene, -
carotene and trans-β-apo-8'-carotenal were purchased from Sigma-Aldrich (Chemie,
Steinheim, Germany). All the solvents used for extraction were of analytical grade and
were obtained from Merck (Darmstadt, Germany). All solvents used for HPLC analysis
(acetonitrile, ethyl-acetate and methanol) were of HPLC grade and were obtained from
Merck (Darmstadt, Germany). Ultrapure water was obtained from a SG-Ultra Clear
water purification system (SG Water Company, Barsbüttel, Germany). Standard
solutions were obtained by dissolving pure compounds in acetonitrile-methanol-ethyl

82
acetate (60:20:20, v/v/v) containing butylated hydroxytoluene (BHT) (1% w/v) and the
solutions were stored in brown flasks at 4oC.
Samples
The eggs used to develop the method were from hens fed a diet containing
waste from tomato processing. Egg yolks were separated manually from their
o
respective whites, then homogenized in a blender and stored at −20 C until carotenoid
analysis. All samples were analysed within three days after extraction. Before analysis,
samples were allowed to achieve room temperature.
Instrumentation
HPLC analyses were performed on a Finningan Surveyor Plus system
(Thermo Electron Corporation, San Jose, CA) including a vacuum degasser, a
Surveyor Plus LCPMPP pump, a Surveyor Plus ASP autosampler and a PDA5P diode
array detector with 5 cm flow cell and with Chrom Quest 4.2 system manager as data
processor. Separation was achieved by a reversed-phase Hypersil Gold C18 column
(5 mm particle size, 250 × 4.6 mm) provided by Thermo Electron Corporation.
Extraction of carotenoids
Carotenoids were extracted from 0.5 g sample with 10 mL of petroleum
ether:methanol:ethyl acetate (1:1:1, v/v/v) containing 0.1% butyl hydroxytoluene (BHT)
by homogenizing for 5 min at 2500 rpm using a Vortex homogenizer. The sample was
centrifuged for 6 min at 6000 rpm and the supernatant was collected. The residue was
extracted following the same procedure until the supernatant was colorless. The
combined supernatants were washed by adding 10 mL of 5% NaCl solution, mixing
vigorously and incubating for 30 min until two layers were separated. The upper layer
was collected, evaporated to dryness under N2 flow and then re-dissolved in 2 mL of
acetonitrile:methanol:ethyl acetate (60:20:20, v/v/v) containing butylated
hydroxytoluene (BHT) (1% w/v). The final solution was filtered through 0.45 µm
membrane filters for HPLC injection.
Chromatographic conditions
The mobile phase was filtered through a 0.45 µm membrane, and degassed
ultrasonically prior to use. The most suitable mobile phase system comprised of
acetonitrile:methanol (95:5, v/v) (A), acetonitrile:methanol:ethyl acetate (60:20:20,
v/v/v) (B) and water (C) with the following gradient: 96% A and 4% C in the beginning,
maintained for 10 min, changed linearly to 100% B in 13 min, maintained 5 min and
returned to 96% A and 4% C in 2 min. The C18 column resolved a total of seven
carotenoids within 35 min. The mobile phase flow rate was 1.5 mL/min. The column
o
temperature was 20 C and the absorbance was read at 450 nm.
The standards were diluted appropriately to prepare standard curves and were
subjected to HPLC-DAD analysis as described above. Standard curves of each
carotenoid were produced by plotting the relative peak areas at 450 nm against the
carotenoid concentrations. Identification was done by comparing the retention times
with those obtained with a mixed standard solution, by spiking of authentic standards
and comparing the spectral data obtained by DAD with reported values. Quantification
was performed using Chrom Quest 4.2 software by comparing peak area with standard
reference curves.

83
 Method validation
Linearity of the detector response was tested by preparing five mixed
calibration solutions. The calibration curves were established by plotting peak area
versus concentration of the analytes. The correlation coefficients were used as the
measure of linearity. The limits of detection (LOD) and quantification (LOQ) were
determined by calculating the concentration of the analytes in mg/L at a ratio of signal-
to-noise (S/N) of 3 and 10, respectively. The precision of the method was confirmed by
repetitive analyses, calculating the average relative standard deviation (RSD) for six
replicate determinations. For the recovery test, standard solutions of carotenoids were
added to the extracts of egg yolk at two concentration levels and then analysed by the
HPLC method for three times. The recoveries for the seven carotenoids were then
calcultated as ratio of the measured concentration to the spiked concentration and
were expressed as a percentage.

RESULTS AND DISCUSSION

The separation of carotenoids using a C18 column was optimized by testing
different solvents and different gradient profiles of co-solvents, temperature and flow
rate. In Figure 1 the separation of the eight carotenoid compounds in the standard
mixture solution was given. The HPLC method thus developed was evaluated in egg
yolk samples by the evaluation of linearity, reproducibility, precision, recovery and
detection limit (LOD).
Linearity
The linearity of the method was evaluated according to area response. The
use of a diode array detector allowed us to confirm the identity of the peak not only by
its retention time, but also by the overlay of the UV-VIS spectra with a standard. The
calibration graphs for carotenoids were produced by injecting standard solutions in the
range 2–20 mg/L. Each point of the calibration graph corresponded to the mean value
obtained from three independent area measurements. Retention times, regression
equations and correlation coefficients for carotenoid compounds were summarised in
Table 1.
As can be deduced from the calibration parameters shown in Table 2,
the correlation coefficients of linear regression analysis were always above 0.997.
Therefore, the external standard method was used for the quantitative analysis.
Reproducibility and detection limit
Chrom Quest software was used to test peak area and retention time
reproducibility. A standard mixture solution of 10 mg/L of each analyte was analysed
six times to determine the reproducibility of the peak area and retention time under the
optimum conditions in this experiment. The relative standard deviations (RSDs) for
retention time were between 0.090 and 0.491% while for peak area the RSDs were
between 0.339% and 1.211%. These values indicated the stability of the method in
terms of peak area and retention time. The detection limits were between 0.041 and
0.752 mg/L for seven carotenoid compounds.

84
 Table 1
Calibration results for determining seven carotenoid compounds in egg yolk
RT Regression equation Concentration Correlation Detection
Carotenoid
(min) Intercept Slope range coeffiicient limit
compounds 2
(mg/L) r (mg/L)
Astaxanthin 3.71 -0.02353 8.19028 e-006 1.2-15 0.9997 0.398
Lutein 4.95 0 7.68226 e-006 0.9-15 0.9996 0.259
Zeaxanthin 5.20 0 6.81732 e-006 0.8-15 0.9994 0.236
Canthaxanthin 6.31 0 8.47711 e-006 2.5-20 0.9990 0.752
Trans-β-apo- 6.51 0 5.34553 e-006 1.6-15 0.9997 0.490
8'-carotenal
Lycopene 17.29 0 4.46802 e-005 1.8-15 0.9970 0.522
β-Carotene 23.27 0 1.28077 e-005 0.2-20 0.9967 0.041

Precision and stability

To test the precision and stability of the HPLC method, a sample solution
(extract) was analysed by six repeated injections and the analysis was repeated over
three days. For retention time, RSD values were between 0.081 and 0.755%, while for
peak areas, RSD values were from 1.692 to 4.815%. These results show that the
HPLC method has good precision and stability.

Figure 1. Chromatogram of an external standard mixture at  = 450 nm

85
 Figure 2. Chromatogram at  = 450 nm of an egg yolk extract

Sample analysis and recovery

Egg yolk extracts were injected directly and separated under the optimum
condition mentioned earlier. A typical chromatogram in the extract of egg yolk is shown
in Figure 2. The calculated contents of the eight carotenoids were given in Table 3.
The recovery experiments of the seven carotenoids were performed by adding
carotenoid standards to the extract of egg yolk, which were treated according to the
procedure described in Section 2.4 for six times. The recoveries for the seven
carotenoids were between 97.27 and 99.98% (Table 2).
Table 2
Contents and recoveries of seven carotenoids in the extract of egg yolk (n = 6)

Component Content (mg/L) RSD (%) Amount Recovery (%)

added (mg/L)
Astaxanthin 0.00 2.53 1.0 93.60
2.5 98.56
Lutein 0.92 4.81 1.0 97.19
2.5 100.08
Zeaxanthin 0.82 1.69 1.0 98.22
2.5 99.86
Canthaxanthin 0.14 3.26 1.0 93.69
2.5 101.88
Trans-β-apo-8'- 0.01 1.94 1.0 92.96
carotenal 2.5 95.04
Lycopene 0.35 2.93 1.0 103.61
2.5 101.78
β-Carotene 0.02 1.79 1.0 98.87
2.5 96.89

86
 CONCLUSIONS
The RP-HPLC method developed here represented an excellent technique for
simultaneous determination of seven carotenoids in the extract of egg yolk, with good
sensitivity, precision and reproducibility. The method gives a good resolution among
analytes with a relatively short analysis time (35 min). The method can be used as
quality control of carotenoids in egg yolk and will play a reference role in the
understanding the influence of the diet and environment on the carotenoid content of
the egg yolk.

ACKNOWLEDGEMENTS
This work was supported by a grant of the Romanian National Authority for
Scientific Research and Innovation, CNCS/CCCDI – UEFISCDI, project number PN-III-
P2-2.1-BG-2016-0019, within PNCDI III.

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Application of a UV–vis detection-HPLC method for a rapid determination of lycopene
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Res. Int. 44, 1284–1291.
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Handelman G., Nocolas R.J. (2006). Consumption ofone egg per day increases serum
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 Singh V.P., Pathak V., Akhilesh K.V. (2012) Modified or enriched eggs: A smart
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88
 Series:  Biology
ANNALS OF THE  Horticulture
 Food produce processing
UNIVERSITY OF CRAIOVA technology
Vol. XXII (LVI II ) - 2017  Environmental engineering

THE INFLUENCE OF APPLYING SUMMER PRUNING OPERATIONS ON

THE QUANTITY AND QUALITY OF CARDINAL VARIETIES

Costea Dorin Constantin1*, Cichi Daniela Doloris1, Căpruciu Ramona1

1
University of Craiova, Faculty of Horticulture A.I.Cuza Street, No. 13, Craiova,
*
Correspondence author. E-mail: [email protected]

Key words: table grapes, summer pruning operations, quality.

ABSTRACT
Summer pruning works and operations apply during the vegetative cycle of grape vines
for the management of growing and fruit bearing seasons, for the improvement of quality,
appearance of grapes and earliness of production. They allow a proper ventilation and
illumination of stock elements, thus reducing diseases and improving the production quality. The
paper shows the results obtained by applying the works and operations to Cardinal table grape
varieties grown in Simnicu de Sus viticultural centre and their influence on the quality of grape
production.

INTRODUCTION
One may say that table grapes must first be appealing to the eye and only then
can be tasted. This is why the appearance is very important in case of table grapes,
holding a decisive influence on consummers‟decision.
Summer pruning works and operation include several plant-growing measures
to be performed during the vegetative cycle of grapevines.
Summer pruning holds greater importance in case of table grape varieties,
owed to its positive influence over grapes quality. When summer pruning works are
carefully and timely applied, their efficiency is higher. Summer pruning must be
performed reasonably considering that leaves are useful body parts for plants and that
each variety has a specific leaf area / output ratio. For this reason, both nationally and
internationally, several research studies have been conducted to determine which
summer pruning operations fit the biological features of varieties best for the
determination of perfect execution time and work intensity. Amongst the authors who
conducted such studies are the following: Acimovici D (2016), Cichi Daniela, Costea D
(2008), Costea D.C et al. (2013), Di Lorenzo. et al. (2011), Gatti M. et al (2015),
Olteanu I. et al. (2002), Palliotti A, Poni S (2011), etc.

MATERIAL AND METHODS

Research studies were conducted on Cardinal table grapes varieties in a
vineyard located in Șimnicu de Sus viticultural centre. The experiment was focused on
establishing the best choices of summer pruning operations, as well as their best
execution timing in order to get high quantity and quality levels of production.
In line with the topic proposed, the observations and determinations were
focused on monitoring climate factors for the evaluation of the favourableness of the
89
study year and the determination of the effect of summer pruning use on the quality
and quantity of Cardinal varieties.
The summer pruning applied was evaluated by analyzing biochemical indexes
(content of sugars, organic acids, glucoacidimetric index) and productive indexes
(grape berry weight, yield per trunk).
The options experimented were several combinations of summer pruning
operations, such as shoot thinning and hedging, partial leaf removal, inflorescence
thinning and trimming. The research methodology used is the one specific for this field.

RESULTS AND DISCUSSIONS

Multi-annual average climate data for the Simnicu de Sus viticultural centre are
shown in the graphs below:

Graph. 1 – Multi-annual average monthly precipitation and rainfall days

(by www.meteovista.co.uk/Europe/Romania/Simnicu-de-Sus/)

Graph. 2 – Multi-annual average monthly precipitation and rainfall days

(by www.meteovista.co.uk/Europe/Romania/Simnicu-de-Sus/)

Graph. 3 Multi-annual average monthly high and low temperatures

(by www.meteovista.co.uk/Europe/Romania/Simnicu-de-Sus)
90
 An analysis of the multiannual data indicated in the graphs above shows that
in most of the years Șimnicu de Sus viticultural centre met the climate conditions
required for high quality grape yields.
Table 1
Climate Data during the mature ripening – Șimnicu de Sus
(by www.accuweather.com/ro/ro/simnicu-de-sus)

Date Precipitation High temp. Low temp.

(mm) (° C) (° C)
16 July 25 27 19
1 August 13 36 18
24 August 16 30 16
19 September 14 32 16

The climate data during the mature ripening process (July-September) shows
a lack of precipitation during 2016 year. Table 1 shows that during the two-month
period there were only 4 rainfall days (over 10 mm), resulting in the early production of
grapes which reached their maturity peak about 2 weeks earlier.

Table 2
The influence of shoot thinning on the production quality of Cardinal variety

Finished Sugar
Production Acidity Glucoacidimetric
Production content
Options (kg/stock) (g/l H2SO4) Index
(%) (g/l)
V1 Total extra shoot 1,87 73 133 5,1 26,0
thinning
V2 50% extra shoot 1,75 79 142 4,9 28,9
thinning
V3 20% extra shoot 1,72 75 139 5,0 27,8
thinning
V4 No summer pruning 1,66 74 131 5,2 25,1
operations
(witness)

The influence of shoot thinning on the quality of production is shown in Table

2. In case of Cardinal varieties the production of V4 (witness) was lower compared to
other options. The biggest production was 2.46 kg / stock and resulted from V1 (total
extra shoot thinning), where the highest content of sugars was also determined. Out of
the shoot thinning options, lower results came out from the 20% extra shoot thinning
operation (Table 2).
Partial defoliation consisted in removing old leaves placed in front of grape
berries to allow their sun exposure, which is so important for their ripening, the stock
ventilation and the easiness of applying plant-growing treatments.
The positive effects of old leaves removal on the quality of table grapes are:
the uniform colouring of grape berries in table grape varieties, the increase of sugar
content, the decrease of total acidity by degradation of malic acid under the more
intense action of light.
The influence of partial leaf removal on the quantity and quality of production
of Cardinal variety is shown in Table 3. The best quantity and quality results came out
91
when the leaf removal operation took place after early ripening and the lowest results
were obtained in case of the witness option.
. In terms of quantity, the production of Cardinal variety, as well as sugar
content, were higher in case of variants V1 and V2 vs witness.
Table 3
The influence of partial leaf removal on the production of Cardinal variety

Finished Sugar
Production Acidity Glucoacidimetric
Production content
Options (kg/stock) (g/l H2SO4) Index
(%) (g/l)
V1 1,71 73 147 4,5 32,26
Leaf removal in
early ripening
V2 1,73 79 150 4,1 36,5
Leaf removal after
early ripening
V3 Leaf removal 1,70 75 139 4,8 28,9
before early
ripening
V4 No summer pruning 1,66 74 131 5,2 21,1
operations
(witness)

Shoot hedging allowed the partial removal of the last 5-7 leaves from the shoot
top which were considered to be consummers, due to the fact that they had no more
time left to reach maturity. The shoot hedging also had a positive influence on the
quantity and quality of production. The biggest production came out when the shoot
hedging operation took place in early ripening (1.75 kg/stock), compared to the lowest
production obtained in case of the shoot hedging before early ripening (1.66kg/stock)-
Table 4
Table 4
The influence of shoot hedging on production of Cardinal variety

Production Finished Sugar Acidity Glucoacidimetric

Options (kg/stock) Production content (g/l H2SO4) Index
(%) (g/l)
V1 Shoot hedging in 1,75 73 148 4,6 32,71
early ripening
V2 Shoot hedging 1,70 79 150 4,3 34,8
after early ripening
V3 Shoot hedging 1,63 75 134 5,0 26,8
before early
ripening
V4 No summer 1,66 74 131 5,2 21,1
pruning operations
(witness)

In terms of quality, the finished production was high (79%) when the shoot
hedging was performed after early ripening. The lowest level of finished production

92
was V1 (shoot hedging in early ripening), i.e. 73%, while V4 (no shoot hedging)
indicated 74%.
Table 5
The influence of inflorescence thinning and trimming on production quantity
of Cardinal variety

Finished Sugar
Production Acidity Glucoacidimetric
Production content
Options (kg/stock) (g/l H2SO4) Index
(%) (g/l)
Inflorescence
V1 trimming after 1,71 80 152 4.3 46.42
blooming
Inflorescence
V2 thinning after 1,79 82 148 3.5 54.28
blooming
No summer pruning
V3 1,66 74 131 5,2 21,1
operations (witness)

Inflorescence thinning greatly influences the quality of grapes through better

commercial appearance, uniformity of maturation, colour and berry size, as well as
increase of sugar content. The influence of inflorescence thinning and trimming on the
production quantity of Cardinal variety is shown in Table 5. In case of options where
summer pruning operations took place, production numbers were higher compared to
the witness option.
The lowest production of 1,66 kg / stock came out in case of V3 option (no
inflorescence thinning and trimming), while the highest production of 1,79 kg / stock
was in V2 (inflorescence thinning after blooming).

CONCLUSIONS
Summer pruning operations require costly and complex manual work, which
means timely performance, specific for each variety, depending on their biological
specific characteristics.
When carrying out summer pruning operations, one must bear in mind that
leaves are useful body parts for the plant and that every variety has a foliar surface/
production specific ratio.
Research studies indicate that summer pruning operations led to better results.
However, given the high costs required, turning them into practice must be performed
distinctly:
 the shoot thinning was good results: the ratios were different, depending on
their biological features, which in case of Cardinal varieties, recommend the total extra
shoot thinning;
 in case of Cardinal varieties the partial leaf removal place after early
ripening determined the increase of finished production and sugar content and the
decrease of acidity;
 when performed in early ripening, the shoot hedging had a positive impact
on the quantity and quality production levels;
 the best results for Cardinal varieties were obtained during the
inflorescence thinning and trimming operations.

93
 REFERENCES
Acimovic, D., Tozzini, L., Green, A., Sivilotti, P., and Sabbatini, P. (2016)
Identification of a defoliation severity threshold for changing fruitset, bunch morphology
and fruit composition in Pinot Noir. Australian Journal of Grape and Wine Research,
22: 399–408
Cichi Daniela Doloris, Costea D.C., 2008, Soiuri de viţă-de-vie cultivate şi
cultivabile în România, Editura Arves
Costea D.C., Elena Genoiu, Daniela Doloris Cichi , N Giugea,L.C. Mărăcineanu,
2013, Research studies on the influence of pedoclimatic variations over the quality of
grapes in viticultural areas of Oltenia (Romania) - Proceedings of the 36th World
Congress of vine and wine-Vine and Wine between Tradition and Modernity Bucharest
– Romania 2-7 June 2013, ISBN : 979-10-91799-16-4
Di Lorenzo R, Gambino C, Scafidi P, 2011 Summer pruning in table grape
,Advances in Horticultural Science ,Vol. 25, No. 3, pp. 143-150
M. Gatti, A. Garavani, A. Cantatore, M.G. Parisi, N. Bobeica, M.C. Merli, A.
Vercesi, S. Poni, 2015, Interactions of summer pruning techniques and vine
performance in the white Vitis vinifera cv. Ortrugo Australian Journal of Grape and
Wine Research Volume 21, Issue 1.Pages 80–89
Olteanu I, Daniela Doloris Cichi, DC.Costea, CL. Mărăcineanu, 2002, Viticultură
Specială –Zonare, Ampelografie, Tehnologii Specifice, Editura Universitaria Craiova
Palliotti A ,S. Poni, 2011,Traditional and innovative summer pruning techniques
for vineyard management Advances in Horticultural Science ,Vol. 25, No. 3, Special
issue on summer pruning of woody fruit species (2011), pp. 151-163
*** www.accuweather.com/ro/ro/simnicu-de-sus
***www.meteovista.fr/Europe/Roumanie/Simnicu-de-Sus

94
 Series:  Biology
ANNALS OF THE  Horticulture
 Food produce processing
UNIVERSITY OF CRAIOVA technology
Vol. XXII (LVI II ) - 2017  Environmental engineeri ng

DEVELOPMENT OF VINE-GROWING FARMS IN BULGARIA

1* 1
Dimitrova Daniela , Dimitrov Vladimir
1
Institute of Viticulture and Enology, 1 Каla Tepe Street, Pleven 5800, Bulgaria
* Correspondence author. E-mail: [email protected]

Key words: viticulture, farms, specialization, economic potential

ABSTRACT
Overcoming the negative tendencies, emerged over the past few decades, led to a
drastic decline in the production of wine and table grapes, required detailed analysis of the
factors and conditions determining viticulture development potential. In this context, the study
examined the processes of ongoing organizational restructuring in viticulture, as a whole and
depending on the legal status of the farms covering the period 2003-2013. The dynamics in the
number and average size of farms, the level of specialization in the sector and the farms
economic potential were studied. The obtained analysis results showed that the mechanisms of
the National and Common Agricultural Policy during the previous programming period did not
support sufficiently the restructuring of the sector. The opportunities for stabilizing the production
at the current economic conditions should be sought mainly in the framework of the integration
between grape-growers horizontally or vertically on the overall organization of the food chain.

INTRODUCTION
Regardless of the existing traditions in our country, the socio-economic
importance of viticulture continued to lose its position within the framework of the
overall development of agriculture and rural areas. (Slavova et al., 2011, Popov et al.,
2012, Malamova et al., 2013, Dimitrova et al., 2013, Koteva et al., 2014, Dimitrova and
Dimitrov, 2017). The intensive nature of the production of wine and table grapes under
the current economic conditions – rising production costs, low purchase prices,
insufficient financial support, a limited domestic market and weak competitive positions
on the international market had predetermined the dramatic decline of the cultivated
vineyard areas, reflecting on the output volume. Wine grapes production was reduced
by 41.6% in 2016 compared to 2007 while of the table grapes by 26.2%. The
overcoming of the negative trend and the utilization of the existing potential in the
country for viticulture development (soils, climate, relief, varietal composition) had been
a function of the entrepreneurial interest generated in the sector under the influence of
the organizational, economic and management factors of the business environment
(Risina, 2007, Borisov, 2007, Kirechev, 2013, Borisov et al., 2014). The objective of the
survey was to analyze the situation and to outline the trends in the development of the
organizational and economic structure of viticulture under the conditions of the National
and the Common Agricultural Policy.

MATERIAL AND METHODS

The analyses were based on official statistical data with sources from the
Ministry of Agriculture and Food (MAF), Agricultural Statistics Department and
95
Eurostat. The methods of statistical groupings and index analysis were applied (Petrov
et al.). The number of vine-growing farms; the number of the specialized vine-growing
farms; the vineyard area in the farm; the average size of the vineyard area per farm;
the number of permanent workers on farms specializing in viticulture, the number of
family labor force have been discussed in dynamics. The change in the structure of the
cultivated vineyard areas was monitored according to the physical size of the farms
grouped per size of the utilised agricultural area (UAA) and the economic potential of
production structures, expressed in European size units (ESU) in 2003 and in a
standard output (SO) in 2013.
The trends in the number of agricultural holdings, the size of the cultivated
areas with wine and table grapes varieties and the average area of the vineyards per
farm, depending on its legal status – individuals, sole traders, cooperatives, trade
companies and associations have been found out.

RESULTS AND DISCUSSIONS

Vine-growing farms development has been the result of the complex impact of
many factors - technological, economic and social, with a view of the complex nature of
the production activity and the specificity of the grape and wine market. The data
presented in Table 1 showed that for the period 2003 to 2013 the number of vine-
growing farms had been drastically reduced by 76.9%. The dynamics was similar with
regard to the size of the vineyard areas in the farms where the reported decrease was
33.4%.

Table 1
Changes in the number of farms, the area of vineyards on the farms and the workforce in the
period 2003-2013

No Indicators 2003 2005 2007 2010 2013 2013/

2003,
%
1 Vine-growing farms, 22358 141 330 98 260 87 000 51610 23.1
number 0
2 Vineyard area, ha 68490 52 670 51540 52340 45640 66.6
3 Average size, ha 0.306 0.373 0.525 0.602 0.884 288.9
4 Specialized vine- 22650 15380 14420 18120 12080 53.3
growing farms, number
5 Utilised agricultural 27900 26660 32990 31740 29430 105.5
area (UAA), ha
6 Vineyard area, ha 20900 19660 25880 27780 26640 127.5
7 Average size of the 0.923 1.278 1.795 1.533 2.205 238.9
vineyard area, ha
8 Permanently occupied 36140 28050 28750 35010 25070 69.4
work force in
specialized vine-
growing farms, number
9 Family workforce, 34680 26770 26340 32900 22730 65.5
number
Source: Eurostat (http://ec.europa.eu/eurostat/data/database) and own calculations

The rate of decrease in vineyards grown on the farms was less pronounced
than the reduction in their total number, resulting in an increase in the average area of
the vineyards in a single farm (188.9% for the survey period). In spite of the positive
effect, it should be outlined that the average size of the vineyards on the farms had
96
remained significantly lower than the rate of this indicator in some of the leading
European grapes and wine producing countries and competitors on the Bulgarian
market, such as France (10.3 ha), Spain (6.5 ha) and Italy (2.4 ha). The small size of
the cultivated areas on the farms negatively affected the opportunities for efficient
organization of the whole complex of activities related to the preparation and marketing
of the end product - grapes and wine.
The unfavorable market situation, especially in relation to the purchase prices
of wine grapes, as well as the high production and market risk, the limited opportunities
for investing in expanding, renovating and modernizing the production activity, due to
the lack of sufficient financial resources and difficult access to bank lending, the
shortage of qualified workers have been the main part of the macro- and
microeconomic conditions that predicted the refusal of a large number of farmers to be
involved in viticulture. During the survey period, the number of the specialized vine-
growing farms had dropped down by half however their relative share in the total
number of farms cultivating vineyards had increased from 10.1% in 2003 to 23.4% in
2013. The area of vineyards in the farms specialized in grapes production had
increased – by 27.5%, leading to a higher concentration of the production activity
within an agricultural holding from 0.9 ha in the beginning of the period to 2.2 ha in the
last year under review.
A negative phenomenon in the context of depopulated rural areas had been
the declining number of permanent workforce in the specialized vine-growing farms (by
30.6% in 2013 compared to 2003), as the decrease in the number of the family labor
force, representing almost entirely the permanent employment in the production
structures with specialization in viticulture was even higher – by 34.4%. The data
clearly revealed that the entrepreneurial interest was not sufficiently promoted with a
view of maintaining, expanding, renovating and modernizing the production potential in
the sector.

2013

0 ≤ 2 ha
2010 2 ≤ 5 ha
5 ≤ 10 ha
2007 10 ≤ 20 ha
20 ≤ 30 ha
30 ≤ 50 ha
2005
50 ≤ 100 ha
≥ 100 ha
2003

0% 20% 40% 60% 80% 100%

Source: MAF, Agricultural Statistics Department

(http://www.mzh.government.bg/MZH/bg/ShortLinks/SelskaPolitika/Agrostatistics/Structure_agricultural_holdi
ngs/Results_copy1.aspx)
Fig. 1: Structure of cultivated vineyard areas per farm group according to UAA for the
period 2003-2013, %.

The analysis of the cultivated vineyard area distribution according to the

physical size of the farms showed that by 2013 the highest share of the vineyard area
had the farms with UAA over 100 ha (38.5% of the total area of the vineyards,
decreasing from 40.8% in 2003) (Fig. 1). In absolute terms, there had been a trend
towards a decrease in the area of wine and table grapes varieties in this group of
farms, amounting to 10 403 ha during the survey period. Similar was the dynamics in

97
the development of the indicator in the groups of farms with UAA under 2 ha and from
2 to 5 ha where the reduction amounted to 66.4% and 25.1% respectively. At the same
time, in all other groups of farms there had been an increase in the size of vineyard
area, as it was the highest in the group with UAA from 50 to 100 ha – by 166.4% in
2013 compared to 2003.
In the structure of the cultivated vineyard areas, according to the economic
potential of the farms in 2003, the highest share had the market-oriented farms (over 4
ESU), that grew 49.0% of the vineyards in the country (Fig. 2). The farms of mainly
natural character (up to 1 ESU) included 35.0% of the area of the vineyards while the
share of the vineyard areas in the semi-subsistence farms (with economic size
between 1 and 4 ESU) was 16.0%.

7500 ha;
22500 ha;
16%
49%
33470 ha;
24100 ha;
49%
35%
8210 ha;
18%
4410 ha;
10%
3010 ha; 7%
10920 ha;
16%
<2000 EUR 2000-8000 EUR 8000-25000 EUR
<1 ESU 1-4 ESU >4 ESU 25000-50000 EUR >50000 EUR

2003 2013
Fig. 2: Distribution of the cultivated vineyard areas in the country according to the
economic size of the farms in 2003 and 2013.

The analysis of the vineyard areas distribution according to the economic

potential of the farms, expressed by the standard output indicator (SO), showed that by
2013 still a significant part of the vineyard area belonged to the small farms (up to EUR
2000 SO) – 7500 ha (16.0%). Almost half of the area of wine and table grapes
vineyards was cultivated in farms with high economic potential (SO over EUR 50,000).
That fact raised concerns about the viticulture sustainable development in the medium
and long term, especially given the need for significant investments in the
modernization of the technological process, in the face of a growing shortage of skilled
workers, together with the tangible need for upgrading of the output potential in the
sector, which was impossible for farms with low economic potential.
The development of the vine-growing farms according to their legal status is
illustrated in Table 2 and Table 3. Both in wine and table grapes vine-growing the
share of the individuals, owners of vineyards, continued to be prevalent by 2013 –
98.3% of the total number of wine varieties vineyards and 97.5% of the total number of
table grapes vineyards.
The number of farms had been decreasing in all organizational forms except
for the trade companies. The most severe was the reduction in the farms of the
individuals – by 77.6% in 2013 compared to 2003 in the wine-growing farms and by
81.3% in the table grapes farms. However, their share in the cultivated area was still
high – 45.7% of the area of the wine grapevines and 60.4% of the area of table grapes
vineyards were owned by individuals.
The area of the vineyards had dropped down in the cooperatives (by 76.0% for
the wine grapevines and 58.1% for the table grapes), in the associations (65.4% for the
wine grapevines and 35.4% for the table grapes) and in the farms of the individuals
(45.7% for wine grapevines and 32.5% for the table grapes).

98
 Table 2
Change in the number of farms, cultivated areas of wine grapevine vineyards
and their average size by legal status

Legal status Farms, number Area, ha Average size, ha

2003 2013 2013/ 2003 2013 2013/ 2003 2013
2003, 2003,
% %
Individuals 213438 47709 22.4 35870 19488 54.3 0.17 0.41
Sole traders 727 194 26.7 1676 2603 155.3 2.31 13.42
Cooperatives 285 87 30.5 19024 4565 24.0 66.75 52.47
Trade 205 514 250.7 7376 15864 215.1 36.0 30.86
companies
Associations 64 16 25.0 442 153 34.6 6.91 9.56
Source: MAF, Agricultural Statistics Department

The size of the cultivated areas had been increasing in the farms with the
status of sole traders (by 927 ha for wine vineyards and 129 ha for table grapes
vineyards) and in the trade companies (by 8488 ha for wine vineyards and 306 ha for
table grapes varieties).
Table 3
Change in the number of farms, cultivated areas of table grapes vineyards and
their average size by legal status

Legal status Farms, number Area, ha Average size, ha

2003 2013 2013/ 2003 2013 2013/ 2003 2013
2003, 2003,
% %
Individuals 36131 6764 18.7 2652 1791 67.5 0.07 0.26
Sole traders 137 56 40.8 87 216 248.2 0.63 3.85
Cooperatives 85 23 27.1 1151 483 41.9 13.54 21.00
Trade 33 82 248.5 104 410 394.2 3.15 5.00
companies
Associations 16 12 75.0 99 64 64.6 6.18 5.33
Source: MAF, Agricultural Statistics Department

The steady growth of cultivated areas in the farms with the status of sole
traders and trade companies showed a positive tendency for the development of these
organizational forms in the sector, influenced to a large extent by the existing support
mechanisms for the period 2007-2013, mainly The Rural Development Programme,
Measure 112 “Setting up farms for young farmers”, Measure 121 “Modernization of
agricultural holdings” and Measure 141 “Support for semi-subsistence farms
undergoing restructuring”. The share of these organizational forms in the total size of
the cultivated vineyards with wine and table grapes varieties had not been high enough
to ensure a long-term effect on the overall development and competitiveness of wine
and table grapes production. The existing possibilities for financial support through the
CAP 2014-2020 mechanisms, both through the multi-component nature of direct
payments and through investment support measures and knowledge transfer and
innovation under the RDP, might contribute to strengthening and enhancing the
viability of the agricultural holdings in the sector.

99
 CONCLUSIONS
The results of the analysis of the organizational and economic structure
demonstrated that, the overall restructuring in the viticulture was taking place at a slow
pace – still the highest share was of small-sized farms and low economic potential
holdings. The need for policy measures to stimulate the process of consolidation of
vineyards is tangible. Despite the positive trends related to the increased concentration
of the production and the positive development of some of the organizational forms, it
was disturbing that in the bigger farms the investment interest in viticulture was
decreasing.
The encouragement of the entrepreneurial initiative and its successful
implementation for stabilizing the production and supply of table and wine grapes and
wine in the country was related not only to the utilization of the provided financial
support opportunities for the sector, but also to the realization of the need to integrate
the production activity in the overall organization of the food chain and bringing
together farmers‟ efforts in the form of producer organizations, producer groups or
other network structures.

REFERENCES
Borisov, P., 2007, Competitiveness of Vine-Growing Enterprises in Southern
Region, Agricultural Economics and Management, 52, 5, 9-17.
Borisov, P., T. Radev, V. Koprivlenski, 2014, Investment activity of vine-growing
holdings and secondary effects under the conditions of CAP (Following the example of
the Municipality of Lyubimets), Agricultural sciences, vol. 6, issue 16, 101-108.
Dimitrova, D., V. Dimitrov, 2017, Regional aspects of viticulture and wine
production development in Bulgaria, Journal of Mountain Agriculture on the Balkans,
20 (3), 162-186.
Malamova, N., D. Mitova, P. Yovchevska, Ts, Kovaceva, B. Ivanov, I. Petrova, P.
Kirovsky, D. Toteva, 2013, Effects of the Implementation of the CAP on the
Development of the Bulgarian Agriculture and Food Industry, Agricultural Economics
and Management, 58, 5-6, 14-27.
Popov, R., M. Atanasova, Y. Slavova, V. Mitsov, 2012. The impact of the EU CAP
on Bulgarian agriculture, Agricultural academy, Acad. Publ. House, S., 60 p.
Petrov, V., P. Angelova, K. Slaveva, 2004, Methods of analysis and management
in agribusiness, Abagar, p.302
Risina, M. 2007. Influence of the Common Agricultural Policy of the European
Union over Vine-growing Development, Agricultural Economics and Management, 52,
2, 9-16.
Slavova, Y., Atanasova, M., Kovacheva, T., Koteva, N., Bachev, H., Mladenova,
М., Malamova, N., Petrova, I., Kaneva, К., Mitova, D., Ivanov, B., Mitzov, V., D.
Pantaleeva, 2011, Competitive opportunities of agrarian sector, Agricultural Academy,
pp. 286

100
 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
 Food produce process ing
UNIVERSITY OF CRAIO VA
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

INFLUENCE OF DIFFERENT GIBBERELLIC ACID (GA 3)

CONCENTRATIONS ON THE INTENSITY OF THE MAIN PHYSIOLOGICAL
PROCESSES ON VARIOUS GREEN LETTUCE HYBRIDS

Aurelia Dobrescu*, Gheorghita Hoza, Gabriela Neață, Ana Maria Petre, Monica
Luminița Badea, Mohammed Jasim Mohammed

University of Agronomic Sciences and Veterinary Medicine Bucharest, Faculty of Horticulture

*Corespondence author: [email protected]

Key words: gibberellic acid, photosynthesis, respiration, transpiration, growth

ABSTRACT
The study presents aspects of the intensity of the main physiological processes
(photosynthesis, respiration, transpiration) in some salad hybrids under the influence of growth
stimulating substance – gibberellic acid. The intensity appraisal of these physiological processes
was determined with the LC Pro + instrument directly in the field of experience at the time of
harvesting. Green lettuce belongs to the Compositae family, is an annual herbaceous and
vegetable plant. It is cultivated in all our country areas, on large surfaces, protected gardens or
directly in the field. The administration of gibberellic acid to various green lettuce hybrids had a
stimulating role in the development of their metabolic reactions. The obtained results were
correlated with the growth process of the targeted plants pursuing the weight of their commercial
part. The intensity of physiological processes varies depending on the light intensity,
temperature and giberelic acid concentration applied to selected green lettuce hybrids.

INTRODUCTION
Green lettuce is an annual herbaceous plant, belonging to Compositae family.
It has been cultivated since ancient times, being used in nutrition as well as in
medicine due to its therapeutic effects. Green lettuce leaves are short petiolate, spiral
arranged, covering the apical meristem. The outer leaves are larger with a well-
developed middle nervure. Layout of the leaves differs according to assortment (Burzo
et al. 2005). The growth of leaves is mainly influenced by light, temperature, nutrition
and humidity (Bensink, 1971). The light effect on leaf growth is dependent on both the
active photosynthetic radiation intensity and the light radiation interception respectively
the foliar surface index. Gaudreau et al. (1994) pointed out that light is the main factor
adjusting growth and development in plants. Also the temperature conditions the
growth of green lettuce plants, respectively the apical meristem growth located near
the soil. The temperature limits for the growth of lettuce plants varies between 7 and 24
° C, with an optimum of 18 ° C (Lorenz and Mainard, 1980). Among the internal factors
that have an influence on growth and ripening in plants, hormones are bioactive
substances with a decisive role. The growth hormone concentration changes during
the development period (Burzo et al. 2004). The staple objective of this study is to
highlight the gibberellic acid concentration influence on the intensity of the main
physiological processes (photosynthesis, respiration, transpiration) occurring in various
green lettuce hybrids.
101
 MATERIAL AND METHODS
The researches took place in the first decade of April, following the main
physiological processes: photosynthesis, respiration, transpiration of the mature leaves
of inland origin green lettuce hybrids under the influence of the growth stimulating
substance gibberellic acid (GA 3). The intensity of photosynthesis, respiration and
transpiration processes was determined with the LC Pro + analyzer directly in the
experience field. Photosynthesis and respiration values were expressed in
μmol/CO2/m 2/s and the transpiration values in µmol/H2O/m2/s. Also, the light intensity
2
was situated beetwen 180 and 926 µmol/m /s and the temperature beetwen 10.8 and
19.5º C. The obtained results were correlated with the growth process achieved by the
investigated plants regarding the weight of their commercial part. The analyzed green
lettuce hybrids were: Touareg, Shangore, Alanis, Centore, Analena and Sotalis. The
examined variants outline for each hybrid included the V1 variant representing the
witness hybrid, V2 referring to the plants that were treated with GA 25 mg/L
concentration and V3 representing GA 50 mg/L treated plants. Physiological
researches were performed at the time of harvesting and the data obtained was
recorded in the graphics.

RESULTS AND DISCUSSIONS

According to the data presented in Graphics as far as Touareg hybrid goes
plants belonging to V2 recorded an intensity of 1.65 times greater than V1 and 1.15
higher than V3. Regarding transpiration a remarkable thing is highlighted: the process
unfolds directy dependent upon temperature, according to the scientific literature data.
The highest transpiration intensity is registered at V3, being 1.22 higher compared to
V1 and 1.11 compared to V2. Note that the determinations were performed at different
temperatures, the highest being at V3. As respects the respiration process, a high
intensity was obtained at V2 plants, which demonstrates that gibberellic acid in the
concentration used (GA 25 mg/L) had a stimulating role in the metabolic reactions
carried out in the plants. Therefore, the metabolic reactions enhancement led to the
growth process stimulation, so the growth expressed in plant weight was the most
obvious at V2 plants, being 1.17 times higher than V1 and 1.10 times higher than V3.
Thus, for this hybrid, it is possible to appreciate the efficient application of the GA 25
mg/L concentration (figure 1).

Figure 1. The Intensity of Physiological Processes of Touareg

The Shangore hybrid, analyzed under the same fertilization conditions has the
highest photosynthesis intensity at V2, being 1.20 higher compared to V1 and 1.12
higher compared to V3. The photosynthesis process is also stimulated in this hybrid by
GA 25 mg/L, but we appreciate that photosynthesis is 1.62 times lower than Touareg.
The transpiration process unfolds with a similar intensity, the differences being
102
insignificant, which makes it possible to assume that in this case gibberellic acid does
not change the conduct of transpiration process. The respiration process determined in
this hybrid variants indicates increasing values from V1 to V3, but with reduced
differences. Regarding Shangore hybrid, although there are relatively few differences
in physiological processes, it is observed the influence of GA 25 mg/L on the growth
processes stimulation (figure 2).

Figure 2. The Intensity of Physiological Processes of Shangore

The Alanis hybrid variants indicate the highest photosynthesis intensity at V1,
being 2.1 times higher than V2 and 1.21 higher compared to V3. It was found that GA
50 mg/L concentration applied to V3, accelerated the photosynthesis process by 1.73
times compared to V2. As far as the transpiration process, it is very intense at V3
plants, being 1.36 higher than V1 and 1.16 higher than V2, which shows us that the
water absorption process is stimulated under the action of GA 50 mg/L. The respiration
process records the highest intensity at V1, being 1.76 times higher than V2 and 1.42
times higher than V3. It is noticed that in this hybrid case, V1 plants have the intensity
of photosynthesis and respiration much more intense than those of the plants under
the influence of growth stimulators, yet the commercial weight of the latter is highly
appreciated (figure 3).

Figure 3. The Intensity of Physiological Processes of Alanis

Centore hybrid records various intensities of the photosynthesis process, but
with insignificant differences, which can not give us relevant information about the
gibberellic acid treatment at V2 and V3 plants. As far as the transpiration process
goes, there is registered a high intensity on V1 plants, being 2.51 times higher than V2
and 1.49 times higher compared to V3. This aspect suggests a balanced water regime
for V1 plants. The respiration process records a high intensity at V3 plants, followed by
V2 plants and the lowest at V1 plants. The physiological indicators values are reflected
103
in the increase process, considering that the V2 plants have a clear growth over the V1
(figure 4).

Figure 4. The Intensity of Physiological Processes of Centore

Analena hybrid under the influence of growth stimulating substance GA

realizes the main physiological processes with the highest intensities in V1 plants. This
hybrid shows a commercial weight increase of the fertilized variants, although the
physiological processes are carried out with similar intensities (figure 5).

Figure 5. The Intensity of Physiological Processes of Analena

Researches on Sotalis hybrid revealed that the physiological processes show

insignificant differences beetwen variants. Thus, the photosynthesis process is carried
out with a similar intensity at V2 and V3 plants and 1,006 times higher at V1. As far as
the transpiration goes, we notice the highest intensity in V1 hybrids and the lowest in
V3 plants, which explains the negative influence of GA 50 mg/L stimulator on water
absorption, The respiration process is carried out with a higher intensity at V2, being
1.14 times higher than V1 and 1.65 times higher than V3. From a physiological point of
view, the variants V1, V2, V3 did not record significant differences, however, regarding
the plant growth of the fertilized variants there was a slight increase compared to
unfertilized variants (figure 6).

104
 Figure 6. The Intensity of Physiological Processes of Sotalis

Table 1
Biometric synthesis data of Green Lettuce, 2017

Edible part, g
Hybrid Variant AVG. nob weight at AVG. weight of Unsalable part
harvest time salable nob weight
unfertilized 252 243 9
SOTALIS GA 25 mg/L 294 285 9
GA 50 mg/L 297 278 19
ANALENA unfertilized 298 283 15
GA 25 mg/L 333 323 10
GA 50 mg/L 379 354 25
CENTORE unfertilized 301 274 27
GA 25 mg/L 401 376 25
GA 50 mg/L 350 312 18
TOUAREG unfertilized 338 304 34
GA 25 mg/L 398 352 46
GA 50 mg/L 361 321 40
ALANIS unfertilized 328 289 39
GA 25 mg/L 377 349 28
GA 50 mg/L 388 340 48
SHANGORE unfertilized 336 300 36
GA 25 mg/L 398 373 25
GA 50 mg/L 350 314 36

CONCLUSIONS
Regarding the Touareg hybrid, the efficient application of the gibberellic acid
must be in a concentration of 25 mg/L. As respects Shangore hybrid, although there
are relatively few differences in physiological processes, it is noticed GA 25 mg/L to
have a stimulating influence over the growth processes. In Alanis hybrid case, although
V1 plants have a higher intensity of photosynthesis and respiration than the fertilized
plants, the latter have a more obvious growth. In the matter of Centore, gibberellic acid
applied in a concentration of 25 mg/L is thought to achieve a metabolism that
stimulates the growth process. In relation to Analena hybrid there is a obvious growth
of the fertilized variants, although physiological processes are carried out with similar
intensities at all variants. Regarding Sotalis hybrid, there were no significant
physiological differences in all three variants, however, fertilized plant are observed to
have a slightly increase in plant growth compared to unfertilized ones. As respects
Analena and Sotalis hybrids, GA doesn‟t influence the physiological prcesses and the
50 mg/L use is not justified as the plant growth processes show reduced differences.
105
 REFERENCES
Bensink J.,1971, Mededelingen Landbouw Hogeschool te Wageningen, pp.71,1-
93.
Burzo I., Delian Elena, Dobrescu Aurelia, Voican Viorica, Bădulescu Liliana, 2004,
Fiziologia plantelor de cultură, Ed. Ceres, București.
Burzo I., Voican Vorica Ana, Luchian Viorica, 2005, Fiziologia plantelor de cultură,
vol. V, Ed. Elisavaros București, pp. 242-244.
Dobrescu Aurelia, 2007, Fiziologia plantelor, Ed. Elisavaros, București.
Gallardo M., ș.a, 1995, J.Plant Nutr.18, pp. 437-453.
‟
Lorenz O.A., Maynard D.N., 1980, Knott s Handbook for Vegetable Growers, New
York.

106
 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
 Food produce process ing
UNIVERSITY OF CRAIO VA
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

INFLUENCE OF FERTILIZATION MODE ON PRODUCTION OF SOME

TOMATO CULTIVARS GROWN IN AN ECOLOGICAL SYSTEM

Domocoş Daniela1, Apahidean Alexandru Silviu1, Cărbunar Mihai2, Mariana Bei2,

1* 1
Apahidean Alexandru Ioan , Ştef Adrian Valeriu
1
University of Agricultural Sciences and Veterinary Medicine, Faculty of Horticulture,
3-5 Mănăştur Street, Cluj-Napoca, 400372, Romania,
2
University of Oradea, Faculty of Environmental Protection,
B-dul Gen.Magheru 26,.Oradea, Romania
Corresponding author. E-mail: *[email protected]

Key words: tomatoes, cultivar, fertilization, production, ecological system

ABSTRACT
Tomatoes are used in human diet, fresh or in cooked food (soups, sauces, pots, stuffed
tomatoes, etc.). They are recommended as food in asthenia, inadequacy, chronic poisoning,
congestive conditions, atherosclerosis, vascular diseases, arthritis, gout, rheumatism, azotemia,
biliary and urinary lithiasis, constipation, enteritis. They are industrially used in the production of
tomato paste, broth, canned, simple or spicy juices. In order to achieve economically profitable
productions, organic farming system requires increased attention to crop fertilization. The
experience was carried out during the year 2016, in an ecologically certified vegetable micro-
plant, in Husasau de Tinca, Bihor County. The experimental factors were cultivar and
fertilization. Experimental culture was set up in a polyethylene tunnel in April.

INTRODUCTION
Tomatoes are an important source of many antioxidant properties: carotenoids
(lycopene, β-carotene), flavonoids, phenolic acids (chlorogenic acid, gallic acid) and
ascorbic acid (Hallmann, 2012). They provide valuable protection for the human body.
To prevent oxidative stress a diet based on the consumption of antioxidants is needed
(Horotan, 2014).
Dry matter content of tomato fruits ranges between 5.5-7.5%, 3-4% glucose
and glucose glucose, 1-1.3% antide and 0.5-0, 7% minerals from the fresh substance.
Fresh fruits contain vitamin C (15-30 mg/100g), carotenoids (0.8mg/100g),
vitamins in group B (B1 0.06mg, B2.0.04mg, B6 0.1mg) and P, vitamin K (24 mg/100
g). Tomato fruit has a high content of mineral substances and organic acids (0.3-
0.5%), mainly represented by citric and malic acid (Dinu et al., 2016).
Favorable ratio of carbohydrates to acids gives fruit and tomato juice a
pleasant taste and a refreshing effect.
Mineral nutrition of tomatoes has some particularities that must be taken into
account when fertilizing. Thus, specific consumption varies depending on the system of
cultivation, variety, production, degree of supply of soil with mineral elements (Indrea et
al., 2012).
Tomato assortment is very varied and dynamic, and varieties and hybrids with
superior agroproductive qualities are grown every year and can be grown in different
107
systems. In the assortment there are different fruit cultivars (most of them are red, but
they are also green, brown, black, yellow, pink or even tigrated). The chemical
composition of fruits differs significantly from one color to another (Lagunovschi-
Luchian & Vînătoru, 2016). The lycopene content varies depending on cultivar and
place of culture. For field cultures, content of lycopene varies between 5.2-23.6 mg/100
g, whereas for tomatoes cultivated in greenhouses the content is between 0.1-10.8
mg/100 g (Abushita et al., 2000 ).
Nitrogen nutrition should be guided so as to avoid excess because, especially
under unfavorable conditions (insufficient light, poor supply of K and Mg), causes
excessive foliar growth and delays the formation and maturation of the fruit (Ciofu et al.
2003). If in the conventional agricultural system the possibility of nutrition management
is ensured by the diversity of fertilizers that have complex composition with macro and
microelements, organic fertilization is mainly used for fertilization with organic
fertilizers. Experiments with Cropmax 0.25% provided 12.5% production increases and
by application of Humus the production increase was 17.7% (Dinu et al., 2009). In
accordance with the requirements of Reg.EU 834/2007, the maximum amount of N
from organic and mineral fertilizers must not exceed 170 kg per hectare per hectare
(Stoleru, 2013).
Cultures of tomatoes in protected areas are carried out between 25 March and
15 September, with the aim of producing early productions, beginning with the end of
May, at a lower cost price than the greenhouse production (Stan et al., 2003).

MATERIALS AND METHODS

The experience was carried out in 2016, being located in an ecologically
certified vegetable micro-farm from Husasau de Tinca in Bihor County, in the western
part of Romania.
The purpose of the experience was to determine how some tomatoes cultivars
behave in different fertilization systems, cultivated in a polyethylene tunnel, under the
conditions specific to organic farming. Objectives were to determine the dynamics of
production and total fruit production.
Pineapple, Potiron Ecarlate, Double Rich, Brandywine Pink, Merveille des
Marchés, Caroten de Plovdiv, Estiva F1 and Blue Beautz have been used in the
experience, the seed being produced for organic crops.
Ananas is highly appreciated by consumers for the qualities it has: it has few
seeds, the pulp is firm, dense, sweet, juicy, very fragrant and with an incomparable
flavor. Color of the pulp resembles that of the pineapple. The fruits are large (200-400
g, sometimes they can reach 1 kg) yellow marked in red. It is a late variety, with
indeterminate growth, with leaves similar to potatoes.
Potiron Ecarlate grows indefinitely, with large fruits of 250-600g, red in mixture
with a little yellow, dense, acidic, tasty, very fragrant pulp, ideal for filling.
Double Rich is an cultivar with indeterminate growth, adapted to short
summers or cooler summers. The fruits of 300-500g, are red, firm pulp, with few seeds
and sweet sour taste. They have double the content of vitamin C than the average of
other tomato varieties.
Brandywine Pink is a very old variety considered to be one of the best
varieties, with undetermined growth, with potato-like leaves. The fruits are large (500-
900g), slightly flattened, dark pink, with a very fine flavor. Fruits are sensitive to
cracking.
Merveille des Marchés is a very old variety with undetermined growth, vigorous
and compact growth, with large fruit production. The fruits are red, with pink pulp,
medium-sized (180-200g) with few seeds.
108
 Caroten de Plovdiv is a variety obtained in Bulgaria by researchers from the
Maritza Institute of Vegetable Culture in Plovdiv.
Estiva F1 is an undetermined growth hybrid, producing medium fruits (180-
220g), red in colour, tasty fruits.
Blue Beautz is a variety with undetermined growth that produces fruit weighing
200-250g with dense pulp. Color is a mixture of red and indigo. The fruits are resistant
to cracking and sunburn. Fruits are rich in anthocyan with antioxidant capabilities.
For differentiated fertilization, Agriful (applied to the ground) and Tekamin Brix
(foliar application) were compared to the nonfertilized variant. Combining the two
experimental factors resulted in 24 experimental variants that were placed in three
repetitions.
The experience was placed in a polyethylene tunnel plant (fertlized in autumn
with with 40 t/ha of half-decomposed manure), crop being set up with seedlings
produced in biofuel-heated seedlings shelter. Sowing was done on 6.02.2016.
Seedlings were transplanted in pots of 9x9x9.5 cm and planted in the polyethylene
tunnel on 16.04.2016.
During the production of seedlings, specific works and two treatments with
nettle macerate (to fortify plants and prevent disease attack) have been applied. First
treatment was performed immediately after rising and the second after 10 days of
transplanting. During the vegetation period, the usual maintenance work was carried
out. To prevent disease attack, nettle treatments were performed and in June, two
treatments were done to combat aphids using a mixture of fern macerate and black
soap (vegetable soap made from olive oil). Plant growth was stopped after 8
inflorescences. Fruit picking started in June and lasted until September.
Observations have been made on plant growth, quantitative and qualitative
production.

RESULTS AND DISCUSSIONS

Harvesting of fruits began with some cultivars in June and with others from
July. First ripened fruits were harvested from Double Rich, Brandywine Pink, Merveille
des Marches. By the fertilization on the ground with Agriful, a more favorable harvest
dynamic was ensured for all cultivars. Foliar fertilization with Tekamin Brix ensured a
better dynamics of harvested production compared to unfertilized variants but lower
compared to variants fertilized with Agriful.
Tomato production ranged from 8.48 kg/m 2 at Brandywine Pink and 10.86
2
kg/m at Estiva F1 hybrid (Table 1). As compared to the average of the experience, the
cultivars Ananas, Brandywine Pink and Caroten de Plovdiv produced lower yields, less
than 9.24 kg m 2, differences in production compared to average being significantly
negative.
Potiron Ecarlate cultivar achieved a production of 9.74 kg/m 2, the difference
from the average of the experience being significant and the Estiva F1s cultivar had a
2
production of 10.86 kg/m , the production increase compared to the average of 17,
53% and the production gap, very significant (Table 1).
Unilateral influence of the fertilization system applied in the experimental
culture highlights the favorable effect on the production of the two supplementary
fertilization variants with Agriful and Tekamin Brix respectively, accepted in the
2
ecological culture system. Tomato production was 8.61 kg/m for the additional
2
nonfertilized variant and 10.08 kg/m for the Agriful variant (Table 2).

109
 Table 1
Unilateral influence of the cultivar on tomato production

Cultivar Production Difference to Significance

kg/m2 % culture of the
average (t/ha) difference
Ananas 8.63 93.39 -0.61 0
Potiron 9.74 105.41 0.50 *
Ecarlate
Double 9.11 98.59 -0.13 -
Rich
Brandywine 8.48 91.77 -0.76 0
Pink
Merveille 9.19 99.45 -0.05 -
des
Marchés
Caroten de 8.49 91.88 -0.75 0
Plovdiv
Estiva F1 10.86 117.53 1.62 ***
Blue 9.44 102.16 0.20 -
Beauty
Average 9.24 100.00 - -
DL P 5% 0.45
DL P 1% 0.83
DL P 0.1% 1.14

Table 2
Unilateral influence of the fertilization system on production of tomatoes

Ferilization Production Difference Significance

kg/m2 % to culture of the
average difference
(t/ha)
No fertilization 8.61 100,00 - -
Soil fertilization 10.08 117,07 1.47 ***
with Agriful
Foliar 9.03 104,87 0.42 *
fertilization with
Tekamin Brix
DL P 5% 0.38
DL P 1% 0.94
DL P 0.1% 1.23

The total production of tomatoes was influenced by the cultivar used and
fertilization mode (Table 3).
The additional fertilization on the ground with Agriful provided production
bonuses ranging between 12.43% and 24.42% depending on the cultivar, the
production differences being very significant in the Ananas, Double Rich Brandywine
Pink, Merveille des Marchés, Merveille des Marchés Estiva F1 and Blue Beauty.
Additional fertilization with Tekamin Brix provided lower production yields, below 10%,

110
with the exception of the Blue Beauty cultivar at which the production increase was
11.59% and the production difference compared to the control variant was distinctly
significant.

Table 3
Combined influence of cultivar and fertilization system on the production of organic
tomatoes
Variant Production Difference Significance
Cultivar Fertilization kg/m2 % to culture of the
system average difference
(t/ha)
Ananas No fertilization 7.82 100.00 - -
Agriful 9.73 124.42 1.91 ***
Tekamin Brix 8.34 106.64 0.52 *
Potiron No fertilization 9.25 100.00 - -
Ecarlate Agriful 10.40 112.43 1.15 **
Tekamin Brix 9.59 103.67 0.34 -
Double No fertilization 8.33 100.00 - -
Rich Agriful 9.87 118.48 1.54 ***
Tekamin Brix 9.15 109.84 0.82 *
Brandywine No fertilization 7.75 100.00 - -
Pink Agriful 9.26 119.48 1.51 ***
Tekamin Brix 8.45 109.03 0.70 *
Merveille No fertilization 8.60 100.00 - -
des Agriful 10.10 117.44 1.50 ***
Marchés Tekamin Brix 8.87 103.13 0.27 -
Caroten de No fertilization 8.03 100.00 - -
Plovdiv Agriful 9.13 113.69 1.10 **
Tekamin Brix 8.32 103.61 0.29 -
Estiva F1 No 10.01 100.00 - -
fertilization
Agriful 11.96 119.48 1.95 ***
Tekamin Brix 10.63 106.19 0.62 *
Blue No fertilization 8.54 100.00 - -
Beauty Agriful 10.26 120.14 1.72 ***
Tekamin Brix 3.77 106.49 0.23 *
DL P 5% 0.51
DL P 1% 0.83
Dl P 0.1% 1.16

CONCLUSIONS
On the basis of the results obtained from the research carried out on the
tomatoes grown in polyethyelene tunnel, in the specific conditions in the West of
Romania, using the cultivars Ananas, Potiron Ecarlate, Double Rich, Brandywine Pink,
Merveille des Marchés, Caroten de Plovdiv, Estiva F1 and Blue Beautz, fertilized with
Agriful and Tekamin Brix respectively, the following conclusions were drawn:
- fruit harvest began in June at the Double Rich, Brandywine Pink, Merveille
des Marches;
- Agriful fertilization on the ground has ensured a more favorable harvesting
dynamics for all cultivars;
- unilateral influence of the cultivar revealed tomato production ranging from
8.48 kg / m2 to the Brandywine Pink cultivar and 10.86 kg / m2 in the Estiva F1 hybrid;
111
 - additional soil fertilization, with Agriful assuring production increases ranging
between 12.43% and 24.42% depending on the cultivar;
- Estiva hybrid achieved the maximum production of 11.96 kg / m2 in the
fertilized variant with Agriful.

REFERENCES
Abushita A.A., Daood H. G., Biacs P. A., 2000, Change in Carotenoids and
Antioxidant Vitamins in Tomato as a Function of Varietal and Technological Factors,
J.Agric.Food Chem.,48(6), pp. 2075-2081.
Ciofu R., Stan N., Popescu V., Chilom P., Apahidean S., Horgoş A., Berar V.,
Lauer K.F., Atanasiu N., 2003, Tratat de Legumicultură, Ed.Ceres, Bucureşti, PP. 603-
615.
Dinu M., Cimpoiașu V. M., Bitai G., Ghivercea V., 2009, Influence of stimulating
substances of tomatoes grown in solaria, Buletin USAMV Cluj,Horticulture, 66/1, pp.
415-419.
Dinu M., Soare R., Gruia M., 2016, Bioactive compounds and antioxidant activity
in comercial cultivars of tomatoes (Lycopersicon esculentum L.), SCEM Conference
Proceding, pp.149-156.
Indrea D., Apahidean Al. S., Apahidean M, Măniuţiu D., Sima R., 2012, Cultura
legumelor, Ed.Ceres, Bucureşti, pp.490-500.
Hallman E., 2012, The influence of organic and conventional cultivation system on
nutritional value and content of bioactive compounds in selected tomato type,
J.Sci.Food Agric, 92, pp. 178-185.
Horotan A., Oancea S., Apahidean Al. S., 2015, Effect of fungicide and
acetylsalycilic acid treatments on qualitativ and quantitativ tomato production, Bulletin
USAMV Horticulture, Cluj-Napoca, vol. 70(1-2), pp. 207-213.
Lagunovschi-Luchian V., Vînătoru C., 2016, Legumicultură, Ed.ALPHA MDN,
Buzău, pp.230-234.
Stan N., Munteanu N., Stan T.,2003, Legumicultură, Vol.III, Ed. Ion Ionescu de la
Brad, Iași, pp. 160-170.
Stoleru V., Managementul sistemelor legumicole ecologice, Ed. Ion Ionescu de la
Brad, Iași, pp. 156-162.

112
 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
 Food produce process ing
UNIVERSITY OF CRAIO VA
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

TRENDS IN THE EVOLUTION OF CLIMATIC FACTORS IN THE DEALUL

BUJORUI VINEYARD UNDER THE CURRENT CLIMATE CHANGES
CONDITIONS
1* 1 1
Enache Viorica , Donici Alina , Tabaranu Gabriel
1*
Research and Development Station for Viticultural and Winemaking Bujoru
Correspondence author E-mail [email protected]

Key words: climate risk, vine, air temperature

ABSTRACT
In the last period of time, the action of climatic factors on vine ecosystems is becoming
increasingly prominent. In addition to the damage caused by extreme temperatures, there is an
increased intensity and increasing frequency of extreme weather phenomena. The knowledge
generated by previous research on the implications of global climate change on the viticultural
ecosystem has been used to move to a superior level of sustainable use of vineyards under the
current climate change conditions. The evolution of air temperature, relative humidity and
sunstroke in the Dealul Bujorului vineyard 1980-2016 during was studied. Climatically
comparatively the periods 1980-2006 and 2007-2016 were analyzed. Observations and
determinations made allow us to assess the evolution of climatic factors under the current
climate changes conditions. It was observed a trend for increased frequency of extreme values
of air temperature and increasing duration of sunshine during in the vegetative period.

INTRODUCTION
Recently, we witness the variation of global climate factors as a result of global
warming caused by greenhouse effect enhancing. Climate change has been
highlighted frequently in recent decades by increasing air temperature, considerable
atmospheric precipitation (rainfall and snowfall) and extreme weather phenomena
(Enache Viorica et al, 2007). The long-term analysis has marked a climate change
trend. Simulations with complex global climatic models indicated that the phenomenon
is determined by both natural and anthropogenic factors (IPCC, 2007). According to
the researches, it is concluded that air temperature may rise by 0,1°C / decade in the
coming decades. A number of international studies have concluded that the average
temperature has risen over the past century. An increase of 0,6°C of the average air
temperature was reported. Obvious trends have also been manifested in the evolution
of the daily minimum temperature that has increased more than the maximum daily
temperature (Easterling et al., 1997).
Based on these considerations, the paper aims to present the tendency of
evolution of some climatic factors in the Dealul Bujorului vineyard, analyzing a series of
data from 1980-2006 taking as reference the decade 2007-2016.

MATERIAL AND METHOD

The research was carried out within the Bujoru Research and Development for
Viticulture and Wine Growing Station. Data on average daily, monthly, and annual
113
average temperatures were processed and analyzed. For the purpose of assessing the
trend of temperature evolution data were collected from the Tg Bujor weather station
(AGROEXPERT system). We analyzed:
- Annual average and vegetation period;
- Monthly average temperature;
- Minimum and maximum annual temperature;
- Temperature of temperatures above 30ºC and below -18,0 ºC;
- Deviation of average monthly temperatures from 2007-2016 compared to
1980-2006;
- Actual insolation (hours);
- Relative humidity (%).
A comparative analysis was carried out between 1980-2006 and the reference
period 2007-2016.

RESULTS AND DISCUSSIONS

The air temperature determines the area of the vine culture, the system of
cultivation, the triggering and the passage of the vegetation phenophases, the quantity
and the quality of the production. The vine is relatively pretentious for heat, cultivating
in areas with an annual average temperature of more than 9ºC. If the average annual
temperature is higher than 10 ° C, quality wine-wine products can be obtained. In
figure 1 shows the average annual temperatures over the two time periods: 1980-2006
and 2007-2016. Annual average temperatures were compared to the mobile average
on 5 year and multiannual average. Different average annual temperatures and
average temperatures during the vegetation period were presented. The 5-year mobile
average indicates a trend of average temperature increases between 1980 -2006 and
a downward trend over the period 2007-2016. Between 1980 and 2016, out of the total
of 27 years, 10 years are below the multiannual average and 12 years below the
multiannual average of the vegetation period. Of the 10 years of the reference period
2007-2016, 5 years are below the multiannual average and 7 years below the
multiannual average of the vegetation period.
From the point of view of the monthly evolution of the temperatures, it is
observed that during the months of March, July, August, November and December, the
average monthly temperature is higher than the average of the reference period 2007-
2016 (figure 2).
T oC
23
21
19
17
15
13
11
9
1983
1984
1985
1986

1991
1992
1993

1998
1999
2000
2001

2006
2007
2008

2013
2014
2015
2016
1980
1981
1982

1987
1988
1989
1990

1994
1995
1996
1997

2002
2003
2004
2005

2009
2010
2011
2012

annual vegetative period

annual mobile media mobile media for vegetative period
multiannual media multiannual media for vegetation
Figure 1 The average annual temperature and the vegetation period in the Dealul
Bujorului vineyard during 1980-2006 and 2007-2016
114
 T oC
25,0

20,0

15,0

10,0

5,0

0,0

-5,0
I II III IV V VI VII VIII IX X XI XII media

average 2007-2016 average 1980-2006

Figure 2 The average monthly temperature in the Dealul Bujorului vineyard between
1980-2006 and 2007-2016

The monthly average temperature during the vegetation period from the
reference period 2007-2016 shows a growth trend in April, May, June and September
with a maximum negative deviation of 0,8ºC / October and a positive maximum
deviation of 0,8 / August compared to the 1980-2006 period (figure 3).

T oC T oC
30,0 1,0
0,8
25,0 0,7
0,4
0,3 0,5
20,0 0,4
0,2 0,2
-0,1 0,0
15,0
-0,5
10,0
1980-2006 -0,5
-0,6
2007-2016
5,0 -0,8
deviation
-1,1 -1,0
0,0
I II III IV V VI VII VIII IX X XI XII
-5,0 -1,5

Figure 3 Deviation of the average monthly temperatures in the Dealul Bujorului

vineyard during 1980-2006 compared to the period 2007-2016
Notice a downward trend for the average temperature to drop in August 2007-
2016 as compared to the period 1980-2006. If from the 27 years of 1980-2006 during
the vegetation period were registered 21 years with maximum air temperatures> 35ºC,
representing 77,8%, in the period 2007-2016, 9 out of 10 years recorded maximum
temperatures of air> 35 ° C, representing 90%.
The frequency of days with maximum air temperatures> 30ºC in relation to the
total number of days for the period 1980-2006 is the maximum in July (37,6%).
Comparing the two intervals, the frequency of days with air temperatures> 30º is higher
115
in April, May, June and July for the period 1980-2006 and lower in August and
September (figure 4).
%
37,66 34,62 35,63
40
31,97
35

30
1980-2006
25 19,20 18,22 2007-2016
20

15
7,29
6,20 4,65
10
3,83
0,32 0,41
5

0
IV V VI VII VIII IX

Figure 4 Frequency of days with air temperatures> 30 ºC in relation to the total number
of days

In the figure 5 shows the evolution of the minimum and maximum annual
temperatures in the Dealul Bujorului vineyard between 1980-2006 and 2007-2016
periods. In the conditions of temperate continental climate, the minimal temperatures
are important for the vine culture. During vegetative rest, the buds of the vine freeze at
temperatures between -16,0° C and -18,0° C. Between 1980 and 2006, in 12 years out
of 27, minimum air temperatures were <-18,0°C, representing 44,5%, compared to the
baseline when it was 6 years, accounting for 60%. The frequency of minimum
temperatures <-18º C in the reference range is increasing compared to the 1980-2006
period.
ToC ToC
45 0

40
-5
35
35,0oC
30 -10

25
-15
20 -18,0oC

15 -20

10
-25
5 Tmax Tmin

0 -30
1980
1981
1982
1983
1984
1985
1986
1987
1988
1989
1990
1991
1992
1993
1994
1995
1996
1997
1998
1999
2000
2001
2002
2003
2004
2005
2006

2007
2008
2009
2010
2011
2012
2013
2014
2015
2016

Figure 5 Evolution of the minimum and maximum annual temperatures in the Dealul
Bujorului vineyard between 1980-2006 and 2007-2016

116
 The light sources of the vineyard are appreciated after the sum of the hours of
effective sunshine during the vegetation period (real sunshine - ∑rs). High values of
this indicator favor the accumulation of anthocyanins in red wine grape. From the data
presented in figure 6, light sources in the 2007-2016 reference range are increasing
over the 1980-2006 period, with a surplus of 212.4 hours, especially in July, August
and September.
Rs hours
340,0

290,0

240,0

190,0

140,0

90,0

40,0

-10,0

-60,0
I II III IV V VI VII VIII IX X XI XII

average 2007-2016 average 1981-2006 deviation

Fig. 6 The evolution of real sunstroke in the Dealul Bujorului vineyard between 1980-
2006 and 2007-2016

The growth and fructification processes of the vine are carried out in optimum
conditions with air humidity of 60-80%. For the whole analyzed period, the relative air
unity is within normal limits, with the exception of July and August 2007-2016, when
the monthly average is lower due to high temperatures and to low rainfall (figure 7).
RH %
100,0
80,0
60,0
40,0
20,0
0,0
-20,0
I II III IV V VI VII VIII IX X XI XIIaverage

average 2007-2016 average 1981-2006 deviation

Figure 7 The evolution of real sunstroke in the Dealul Bujorului vineyard between
1980-2006 and 2007-2016

CONCLUSIONS
Comparing the 1980-2006 period with the reference period 2007-2016, it was
found that:
1. The 5-year moving average indicates a tendency to increase average air
temperatures between 1980-2006 and a downward trend over the period 2007-2016,
both on an annual basis and during the growing season.
117
 2. There is a downward trend in the average temperature in August 2007-
2016 compared to the period 1980-2006.
3. The frequency of days with air temperatures> 30,0ºC is higher in April, May,
June and July for the period 1980-2006 and lower in August and September compared
to 2007-2016.
4. The frequency of the years with maximum temperatures> 30,0°C increased
in the reference range to 90% compared to the period 1980-2006 (77,8%).
5. From the 27 years of 1980-2006 during the vegetation period were
registered 21 years with maximum air temperatures> 35,0ºC, representing 77,8%, in
the period 2007-2016, 9 out of 10 years recorded maximum temperatures of air > 35,0
° C, representing 90%.
6. The frequency of minimum temperatures <-18,0ºC in the reference range is
increasing compared to the 1980-2006 period.
7. Light sources are rising in the 2007-2016 reference range.

REFERENCES
Easterling DR, Horton B, Jones PD, Peterson TC, Karl TR, Parker DE, Salinger
MJ, Razuvayev V, Plummer N, Jamason P, Folland CK., 1997, Maximum and
minimum temperature trends for the globe, Science 277: 364–367.
Enache Viorica, 2007, Researches Regarding the Tendency in Some climatic
factors evolusion from Moldavia south hilly region with reference to viticultural
plantation, Lucrări ştiinţifice, Anul L, vol. 1 (50), Seria Horticultura, Editura Ion Ionescu
de la Brad, p.639-642.
IPCC, 2007, Climate change, 2007, The physical science basis, Contribution of
working Grup I tothe fourthassessment raport of the IPCC, Cambridge UP.

118
 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
 Food produce process ing
UNIVERSITY OF CRAIO VA
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

USE OF EXTRACTS FROM RED GRAPE POMACE IN VINE PROTECTION

1 2
Fîciu Lidia *, Bădulescu Adriana
1*
Research and Development Institute for Viticulture and Enology, Valea Călugărească, Romania
2
Research and Development National Institute for Biotechnologies in Horticulture Stefanesti Arges
* Correspondence author. E-mail: [email protected]

Key words: grape pomace, polyphenolic compounds, grapevine downy mildew

ABSTRACT
The aim of this study was to investigate the efficiency of the treatment with Cu 2+
(chelatic) complexes and extract of grape pomace in the protection of the vine. Grape pomace
was obtained by extraction from three black varieties. Cu 2 + complexes were obtained based on
polyphenol compounds separated from red grape pomace. The causal agent pathogen was the
downy mildew produced by the Plasmopara viticola fungus and the treatments were performed
in the field conditions. Three control variants have been tested: classical treatment against
downy mildew with synthesis and contact products, treatment with copper chelate in aqueous
solution 0.1% and treatment with 0,2% aqueous solution of grape pomace. As a result of the
observations made in the case of chemical treatment, the degree of attack was only 9%,
followed by the solution treated with Cu 2 + (chelatic) complexes, respectively 20% and the
treatment with extract of grape pomace, 27%.

INTRODUCTION
The downy mildew is produced by the Plasmopara viticola fungus and is the
most dangerous disease because it attacks all the green organs in formation, causing
very great damage to the culture (Tomoiagă Liliana 2006, Dagostin et al. 2011, La
Torre et al. 2014). Worldwide there are few studies on the use of extracts from grape
pomace to naturally stimulate the resistance of plants to the attack of cryptogamic
diseases. Extracts from grape pomace alter the morphogenesis of parasitic fungi such
as Plasmopara viticola, Uncinula necator, Botrytis cinerea, preventing spore
germination, granulating conidia cytoplasm, destroying plasma membranes, etc.
(Woods et al. 1995). The research by Sackenheim et al. (1994) under controlled
conditions highlighted the positive effect of the extract from grape pomace in reducing
the attack of the Plasmopara viticola fungus on the leaves. The possibility of using the
extract from grape pomace in the control of cryptogamic diseases in vines is of interest
for organic viticulture since 2006, the use of copper products in the control of manure
in the vine was restricted at European Union level living, these products being
accepted in organic viticulture.

MATERIAL AND METHODS

The study was conducted at the Research and Development Institute for
Viticulture and Enology, Valea Călugărească in the 2015-2016 period. The
experimental device was located within a plot of cultivated Merlot / SO4 variety, with
119
planting distances of 2.0 x 1.0 m, located on wide terraces, under the conditions of a
brown-red mollusc soil. The way of soil maintenance was that of a black field. Three
variants of control have been tested: V1-classical treatment against downy mildew with
synthesis and contact products (Dithane M-45, Acrobat Mz 69 WG and Kocide 2000),
V2-treatment with copper chelate in aqueous solution 0.1% and V3-treatment with 0.2%
aqueous solution of grape pomace. The control variant was untreated (V1). Cu2+
(chelatic) complexes were obtained by Polytechnic University of Bucharest, based on
extracts rich in polyphenolic compounds obtained from red grape pomace. The
application of the products was carried out during the vegetation period in the
phenophases BBCH11 (first leafs), BBCH16 (six leafs), BBCH55 (before flowering),
BBCH71 (and of flowering) and BBCH73 (the grapes reached 30% of the size), with
the vermorrel. After each treatment, observations and determinations were made to
record the degree of attack on leaves and grapes and to determine the efficacy of the
products used to combat the Plasmopara viticola fungus. The rating of attack is
expressed by intensity and frequency (GA% = FxI / 100), and these values were the
basis for interpreting the obtained results.

RESULTS AND DISCUSSIONS

Grape pomace was obtained by extraction from three black varieties: Burgund
mare, Feteasca neagra and Merlot.
Physico-chemical characteristics of fresh grape pomace were performed (table
1). Fresh grape pomace presented a humidity of 6.70-7.73 g/100 g and an acid pH.
Sugars are present in quantity between 0.6-1.19 g/l, while the amount of sugars in
grapes had values ranging between 211 and 230 g/l.
Table 1
Physico-chemical characterization of fresh grape pomace
Parameters The raw material-grapes for red wines
Burgund mare Feteasca neagra Merlot
Moisture (g/100g) 7.73 ± 0.07 7.37 ± 0.17 6.70 ± 0.07
pH 3.68 ± 0.04 3.90 ± 0.03 3.67 ± 0.03
Sugar (g/l) 0.46 ± 0.06 0.66 ± 0.09 1.19 ± 0.18
Total acidity (g/l
3.9 ± 0.30 4.6 ± 0.25 4.9 ± 0.35
tartaric acid)
The ashes 7.99 ± 1.05 7.53 ± 1.03 6.56 ± 0.64

The obtaining of aqueous extract from grape pomace has been carried out in
the following manner: 100 g marc pinched and mountainous, has been named in 1000
ml ethyl alcohol 70% and mixed using a shaker for 2 hours at 100 rpm. The resulting
extract was centrifuged for 30ʼ at 3600 rpm, then passed the rotoevaporator at 80°C,
separating the alcohol extract. The extract was brought to 100 ml with distilled water
(I). In order to correct the acidity extraction (which must have a neutral ph) was carried
out and a buffer solution which consisted in: 11.9 g disodium phosphate, Na2HPO4
solution • 2H2O dissolved in 1000 ml of distilled water and 0.9 g potassium dihydrogen
phosphate KH2PO4 solution dissolved in 100 ml of distilled water. 970 ml solution of
disodium phosphate interfered with 30 ml of potassium dihydrogen phosphate solution,
resulting the buffer solution (II). The final extract was obtained through a mixture of 100
ml of extract with distilled water (I) and with the buffer solution (II).
With regard to the climatic conditions, the level of rainfall was very low during
the winter months, when it grew at 0.2 mm in December compared to 44.5 mm

120
multiannual average, but has become surplus in the spring (87.6 mm versus 35.3 mm
multiannual average recorded in March) and in July, favoring the development of
diseases, most notably downy mildew. In the warmest months (June, July and August)
have registered 81.8 mm, 70.2 mm and 78.2 mm. Evolution of average values of
rainfall in the period January-September 2016 is represented in Figure 1. The amount
of the average temperature °C in January-February period was between - 4.3 ° C in
January and 169.3 ° C in July (Figure 2).

Figure 1. The average rainfall Figure 2. The average temperature°C

January-September January-September

Analyzing the data collected in conjunction and with the climatic conditions in
the period of vegetation of vineyards that were particularly favourable to the attack of
downy mildew on leaves, inflorescences and grapes, the first symptoms of leaf downy
mildew attack have been observed from 15.06.2016. Due to the hot and humid climate,
the evolution of the disease was fast so that the degree of attack in untreated (control)
was quite high, 43%. In the case of chemical treatments the degree of attack was only
9%, followed by the solution treated with Cu2+ (chelatic) complexes, respectively 20%
and the treatement with extract of marc, 27%.
Due to the climatic conditions in the period of vegetation of vineyards, which
were very favourable to the attack of downy mildew, the treatment with copper
chelates, but especially with the pomace extract were not effective enough to halt the
onslaught of downy mildew, the reason for that especially young grapes were partially
affected, which has led to a reduction in the production of grapes.

Figure 3. The evolution of the attack of downy mildew (GA%)

121
 CONCLUSIONS
Compared to the treatment with chemicals, the degree of attack was double in
case of treatment with Cu2+ (chelatic) complexes and triple in case of treatment with
extract from grape pomace, but much less than in untreated variant, when the loss of
production was 100%. In the case of these variants, when the weather conditions are
very favourable to the attack of downy mildew (warm and wet weather) are
recommended complementary 1-2 treatments with systemic products (especially in the
phenophases a flowering and the increase of grape berries).

REFERENCES
Dagostin S., Schärerl H.J., Perto T., Tamm L., 2011, Are there alternatives to
copper for controlling grapevine downy mildew in organic viticulture? Crop Prot., 30,
pp. 776-788.
La Torre A., Mandalà C., Pezza L., Caradonia F. and Battaglia, 2014, Evaluation
of essential plant oils for the control of Plasmopara viticola, Journal of Essential Oil
Research.
Sackenheim R., Weltzien H.C., Kast W.K., 1994, Effects of microflora composition
in the phyllosphere on the biological regulation of grapevine fungal diseases. Vitis
33.235-240.
Tomoiagă Liliana, Bolile şi dăunătorii viţei de vie prevenire şi combatere, 2006,
Edit. Mediamira, Cluj-Napoca,.
Woods J.A., Hafield J.A., Pettit G.R., Fox B.W., and Mc Gown A.T., 1995, The
interaction with tubulin of a series of stilbenes based on combretastatin A-4.Br. J.
Cancer 71, pp. 705-711.

122
 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
 Food produce process ing
UNIVERSITY OF CRAIO VA
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

CHARACTERIZATION OF SPONTANEUS FLORA AND INSECTS IN VALEA

CĂLUGAREASCĂ VITICULTURAL ECOSYSTEM

Fîciu Lidia1*, Bădulescu Adriana2, Brîndușe Elena1

1*
Research and Development Institute for Viticulture and Enology, Valea Călugărească, Romania
2
Research and Development National Institute for Biotechnologies in Horticulture Stefanesti Arges
* Correspondence author. E-mail: [email protected]

Key words: flora, harmful insects, useful insects, ecosystem.

ABSTRACT
The mail goal of this paper was to invent the spontaneus flora and insects (useful and
harmful) in Valea Calugareasca viticultural ecosystem. Flora and fauna represent a natural
heritage, and it allows us to appreciate the situation of future threats to the vineyard culture. In
the viticultural ecosystem taken into study the following areas of interest were delimited: the
main crop (vine) in 85.3%, the hedges (5.4%), the canals (4.6%), the rows of trees (0.004%) and
isolated trees (0.002%).
28 floral species were identified following inventory. Of these, the most abundant were:
Agropyron repens and Capsella bursa pastoris. The rare species were: Ranunculus ficaria,
Crocus, Hypericum perforatum. As regards fauna, 61 species of harmful fauna and 138 species
of useful fauna were identified.

INTRODUCTION
The conservation of biodiversity is, nowadays, one of the major issue at the
international level due to intensified human impact on the biosphere. In this context,
the maintenance of biodiversity is necessary not only to ensure life in the present, but
also for future generations, because it preserves the regional and global ecological
balance, guarantee the regeneration of biological resources and maintaining
environmental quality required.
Wild flora and fauna represent a natural heritage of aesthetic, cultural and
scientific value (Ellenberg et al. 1992).
Biodiversity is essential for "services of ecosystems", the services being
offered by nature: adjusting the climate, water and air, soil fertility and the production of
food, fuel, fibre and medicines.

MATERIAL AND METHODS

In a vineyard ecosystem located at The Research and Development Institute
for Viticulture and Enology, Valea Calugareasca, the spontaneous flora, useful and
harmful fauna were inventoried.
Inventory of flora was achieved through collecting and mapping of species with
the help of frames of 1 m 2. Journeys were carried out at regular intervals so as to be
captured on all stages of vegetation and as many species as possible. In order to
identify the plants specialized determinant were used (determinator for flora: “Flora
ilustrată a României” – V. Ciocîrlan, 2009, “Flora R. P. R. ” - Vol. I-XIII,1952-1976).
123
Following the field visits was drawn up the inventory of flora and, also data on plant
fenologia were recorded. The following field observations were made: systematic
database of species, abundance, dominance, phenology. Abundance was calculated
as the total number of species in a given ecosystem area respectively. This indicator is
expressed in absolute value. Depending on abundance, the species may be relatively
rare, common, abundant, very abundant. Dominance was calculated starting from the
abundance as the ratio of livestock species and the amount of other herds in the
studied area. The notion of dominance is an indicator of productivity, showing what
percentage of each species is present in the biocoenosis.
Inventory of harmful and useful entomofauna were made using the Barber
traps, of leaves samples and the fracture technique. Barber traps are the classic
method for collecting ground-moving arthropods (epigee in the case of nocturnal or
diurnal grasshoppers, but also other invertebrates with high activity on the soil, such as
aranas, miriapodes, etc. Six traps per variant were installed in each experimental plot.
Leaf samples were an efficient method of estimating populations at the leaf
level. This method consists in the preleavtion of 50 leaves per variant, collected
randomly once a week (on the same day of the week). They were put in bags with the
batch and date labels, transported to the laboratory, and examined under a
microscope.
The fracture technique was used to evaluate fauna (harmful and useful) in the
experimental variants, completing the other techniques (visual control, buried traps or
Barber). The basic principle of the method is to shake a number of vegetative organs
above a conical or square device (an entomological net).

RESULTS AND DISCUSSIONS

It was necessary to identify and delimitation of habitats being a paramount
relevance for the realization of the programs of monitoring and management of
biodiversity. The microhabitat for vine has been identified as an analysis of its structure
and morphology (Figure 1). The total studied area was 0.129 ha, from which 0.11 ha
the area occupied with vine (main crop).

Figure 1. Analysis of structure and morphology of vineyard microhabitat

124
 Soil type of ecosystem is preluvosol reddish molic (Br MB), the structure was
destroyed towards the poliedric large developed subangular dark brown and wet-dry
dark brown, porous, fine effervescence, moderately friable, with earthworm channels.
The grain size fractions of soil in the ecosystem to study are shown in table 1.

Table 1

Enter the vineyard soil composition from experimental plots (%)

Soil type Depth Coarse Fine sand Dust Clay Textural

(cm) sand < 0.002 class
mm
Preluvosol 0-20 1.76 34.72 20.92 42.60 TT
reddish molic 20-40 12.91 6.41 29.38 51.30 TT
40-60 1.26 21.55 23.69 53.30 AL
TT = clay loamy; AL = clay.

As a result of valuation have been identified a number of 28 species of

spontaneous flora which grow and develop in the wine-growing ecosystem got in this
study. The plants are listed in table 2 after the scientific name and family.
Table 2

Spontaneous flora species reported in vineyard ecosystem

Species Taxon Abundance Dominant

Fam. Malvaceae Althaea officinalis 17 0.839
Fam. Poaceae/ Agropyron repens 258 12.734
Gramineae
Fam. Amaranthaceae Amaranthus retroflexus 97 4.787
Fam. Apiaceae Aethusa cynapium 76 3.751
Matricaria inodora 31 1.530
Polygonum convolvulus 98 4.837
Fam. Asteraceae Cirsium arvense 113 5.577
Chrysantheum vulgare 96 4.738
Taraxacum officinale 82 4.047
Fam. Rubiaceae Galium aparine 215 10.612
Galium verum 36 1.776
Fam. Caryophyllaceae Stellaria media 49 2.418
Fam. Lamiaceae Lamium purpureum 130 6.416
Salvia pratensis 10 0.493
Fam. Plantaginaceae Plantago lanceolata 158 7.798
Fam. Urticacea Urtica dioica 50 2.467
Fam. Umbelliferae Aegopodium podagraria 28 1.382
Fam. Primulaceae Primula officinalis 38 1.875
Fam. Papaveraceae Chelidonium majus L. 42 2.073
Papaver rhoeas 25 1.233
Fam. Ranunculaceae Ranunculus ficaria 5 0.246
125
 Fam. Violaceae Viola odorata 20 0.987
Fam. Fabaceae Lathyrus vernus 32 1.579
Fam. Campanulaceae Campanula patula 25 1.233
Fam. Iridaceae Crocus 5 0.246
Fam. Hypericaceae Hypericum perforatum 8 0.394
Fam. Brassicaceae Sinapis arvensis 22 1.085
Capsella bursa pastoris 260 12.833

As regards fauna, there were identified 61 species of harmful fauna and 138
species of useful fauna (Table 3).

Table 3
Useful and harmful fauna collected
Specify 06-24.06 11-25.07 08-25.08 Total The relative
2016 2016 2016 abundance
%
HARMFUL FAUNA
Gryllus campestris L. - 2 3 5 8.20
Cicada viridis L. 2 2 - 4 6.56
Ceresa bubalus L. - 1 1 2 3.28
Pulvinaria vitis L. 1 1 1 3 4.92
Alopecosa pulverulenta C. - 1 1 2 3.28
Epicometis hirta Poda - 2 1 3 4.92
Apis sp. 1 2 - 3 4.92
Carabus violaceus L. 2 1 - 3 4.92
Eurigaster maura Pul. 1 2 1 4 6.56
Melolontha melolontha L. 2 2 - 4 6.56
Haltica ampelophaga Guer. - 3 - 3 4.92
Lebia humeralis D. - 2 3 5 8.20
Vespa germeanica L. - - 3 3 4.92
Tetranychus urticae Koch 3 - - 3 4.92
Eriophyes vitis Pgst. 2 - - 2 3.28
Anomala solida Erw. 3 - - 3 4.92
Tanymecus palliathus F. 2 1 - 3 4.92
Hyphantria cuneae L. - - 3 3 4.92
Lobesia botrana Den.et Schiff. - - 3 3 4.92
Total harmful fauna 61 100.00
USEFUL FAUNA
Forficula auricularia L. 10 16 8 34 24.65
Blitophaga undata Mull. 1 1 1 3 2.17
Calosoma auropunctata L. 1 1 2 4 2.90
Harpalus pubescens L. - 2 3 5 3.62
Harpalus distiguendus Duft. - - 2 2 1.45

126
Coccinella 7-punctata L. 8 2 2 12 8.70
Propylea 14-punctata L. 5 1 - 6 4.35
Coccinula 14-punctata L. 8 1 1 10 7.25
Thea 22-punctata L 4 2 1 7 5.07
Coccinella septempunctata L. 5 2 1 8 5.80
Scymnus sp. 4 5 2 11 7.97
Philaenus spp. - 7 - 7 5.08
Allobophora caliginosa L. - - 2 2 1.45
Allobophora rosea L. - 3 1 4 2.90
Fridericia bulbosa Rosa - 2 2 4 2.90
Lithobus fornicatus L. 1 1 1 3 2.17
Geophillus sp. - 2 1 3 2.17
Nabis ferus L. 2 2 2 6 4.35
Eurydema ornata L. 1 1 2 4 2.90
Aphodius luridus F. - 1 2 3 2.17
Total useful fauna 138 100.00
Total general 199 -
The relative abundance of 61 30.66
harmful fauna (%)
The relative abundance of 138 69.34
useful fauna (%)

In the first category highlighted the following species: Cicada viridis L., Gryllus
campestris L., Melolontha melolontha L., Hyphantria cunea L., Vespa germeanica L.,
Eurigaster maura Pul. The category of useful species is represented by Forficula
auricularia L, Blitophaga undata Mull., Calosoma auropunctata L., Harpalus pubescens
L., Sciaridae, Chalcidoidea, Ichneumonoidea, Formicoidea.
Copies of each report category and the number of species showed a net
favourable situation for useful fauna. The report overall FU (useful fauna)/FD (harmful
fauna) was 69.34/30.66.

CONCLUSIONS
From the 28 floral species the most abundant were: Agropyron repens,
Capsella bursa pastoris, Galium aparine, Plantago lanceolata and Cirsium arvense.
Rare species were Ranunculus ficaria, Crocus, Hypericum perforatum, Salvia
pratensis and Althaea officinalis.
The most represented families were: Asteraceae - 420 species, Brassicaceae -
282 species, Poaceae/Gramineae - 258 species, Rubiaceae - 10 species. Genres that
include most species are: Capsella - 260 species, Agropyron - 258 species, Galium -
215 species, Genul Plantago - 158 species, Lamium -130 species.
The useful and damaging fauna of the Valea Calugarească vineyard
ecosystem was very varied.

127
 The species that have registered a numerical abundance, important from the
point of view of the protection of the culture were those belonging to the Homoptera
order.

REFERENCES
Сiocârlan V., 2009, Flora ilustrată a României, Bucureşti, Edit. Ceres, p. 1-1141.
Ellemberg H., Weber E.H., Dull R., Wirth V., Werner W., Paulissen D., 1992,
Zeigerwerte von Pflanzen in Mitteleuropa. 2 Aufl., E. Goltze Verlag, Göttingen.
** 1952-1976, Flora R.P România. R.S. România, I-XIII. Edit. Acad. Române.
Bucureşti. 298.

128
 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
 Food produce process ing
UNIVERSITY OF CRAIO VA
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

SOME FRUIT CHARACTERISTICS OF BLACKTHORN

(PRUNUS SPINOSA L.)

Gavrilă Călușaru Florentina Claudia1,*, Ionica Mira Elena, Cosmulescu Sina Niculina2
1,
*University of Craiova, Horticulture Faculty, Doctoral School of Plant and Animal Resources Engineering,
Romania, e-mail: [email protected]
2
University of Craiova, Horticulture Faculty, Department of Horticulture & Food Science, A.I.Cuza Street, 13,
Craiova, Romania; e-mail: [email protected]

Key words: Prunus spinosa L., fruits, physical-chemical characteristics, diversity.

ABSTRACT
This study has been conducted upon 15 genotypes of Prunus spinosa L. growing
wildly in spontaneous flora in South Oltenia Region (Dolj county). Ripen fruits of these genotypes
have been harvested and analysed. Some physical-chemical characteristics have been
observed. Average fruit weight ranged between 1.10 g and 4.20 g and the average pulp mass
ranged between 61.53 and 93.75%. Dried substance content ranged between 17.50% and
25.50% and the values of titrable acidity were established as 1.94; 2.08; 2.78; 2.28; 1.91; 1.27;
1.47; 1.88; 1.51; 1.21; 1.71; 1.74; 1.54; 2.11; 1.61, respectively (g malic acid content/100g fresh
fruit). This study aims at showing that there is a great diversity of Prunus spinosa L. genotypes,
especially regarding the characteristics of fruit evaluated in the same ecological conditions. The
last but not least, these genotypes of Prunus spinosa L. could be very useful not only as a tree-
growing point of view, but also as food additive.

INTRODUCTION
At present, Prunus spinosa L. is the only species of this genre which can be
found in spontaneous form in Romania, especially in steppe areas where it grows in
small bushes (0.3–0.8 m) stretching up to the hills areas. It plays an essential role in
root-sucking in both valley and low hills areas where it grows wildly especially in village
common areas (Cociu et al. 1999). Prunus spinosa L. (blackthorn or sloe) is a species
of flowering plant in the rose family of Rosaceae, subfamily Prunoidae, Prunus genre.
The fruit is 1-1.5 cm globular drupe (Cosmulescu 2014) blueish black, little, short
pedunculate bloomy, ripening in the autumn, covered of prune (Iliescu 2002). The pulp
is green, dense and very astringent to taste. They can be eaten both raw and overripe
(Ghena et al. 1997). These fruits can be a valuable source of natural antioxidants
(Cosmulescu et al. 2017). Even though they are edible and they can be used for
medicinal purposes, it is hard to find products of blackthorn fruits on the market or even
with their additives (Sikora et al. 2013). Blackthorn (Prunus spinosa L.) is used in
phyto-therapy for the treatment of many diseases related to various forms of cough, it
is mild laxative, diuretic, spasmolytic and anti-inflammatory agent. It has anti-septic
effect (due to the presence of tannins) and shows activity against inflammatory of
mucosal layer of the digestive system (Velickovic et al. 2014). Medicinal characteristics
of blackthorn were shown by fruit flowers, bark and root of the plant. Apart from phyto-
therapy, blackthorns are also used in food industry for production of jams and various
129
beverages: liquor, wine, juice, compote and tea (Velickovic et al. 2014). According to
Olszewska and Wolbis (2001) extracts from the flowers have been used as diuretic,
anti-inflammatory, spasmolytic and mild laxative and “blood cleansing” agents, and
their active substances are flavonoids. The fruits are also dietetic products rich in
tannin which have anti-inflammatory and anti-bacterial effects. Apart from, flavonoids,
kaempferol, quercetin and their heterosides can be mentioned among their
constituents. The blackthorn (Prunus spinosa L.) is a thorny deciduous shrub whose
fruits are astringent and they can be eaten fully ripen but it is more common to be
prepared using sugar, honey or alcoholic drinks in order to obtain a digestive liquor
used for its laxative, astringents, diuretic and purgative effects. According to Ruiz-
Rodrigues et al. (2014) blackthorn could be a valuable and cheap source of
antioxidants and she highlights their role in modern and traditional diet. Over a vast
expanse of north-eastern Iberian Peninsula, sloes from blackthorn are macerated to
make a schnapps called “pacharan”. It is a traditional drink of Navarra, Spain flavoured
with the essential oils of anises and sugar. In their study Fraternale et al. (2009)
assessed the antioxidant activity of Prunus spinosa L. growing wildly in Urbino area,
central Italy. Their study revealed that three most representative anthocyanins
contained by Prunus spinosa L. fruit juice (cyanidin-3-rutinoside, peonidin-3-rutinoside
and cyanidin-3-glucoside) are likely to play an important role in its antioxidant
properties. Due to its antioxidant properties, different concentration of smashed
blackthorn can be used for preparing ice-cream. Furthermore, this led to acceptable
effects upon physical, chemical, thermic and sensorial properties of ice-cream. The
use of blackthorn at different concentrations for ice-cream production caused an
acceptable effect on some physical-chemical, thermal and sensory properties of ice-
cream (Kavaz Yuksel 2015). According to Barros et al. (2010) blackthorn contains very
useful bioactive phytochemicals such as phenolics, vitamins (ascorbic acid and
tocopherol) and carotenoids. The combination of bioactive compounds and rich
nutritional composition (high contents in carbohydrates, low contents in fat with
precious contribution of polyunsaturated fatty acids as well as omega 3 and omega 6
fatty acids make them a special food. Marakoglu et al. (2005) showed not only the
chemical and physical properties of blackthorn but also the fact that all materials
contained high amounts of K (18706,98 mg/ kg), Ca (1524.22 mg/ kg), Mg (968.15 mg/
kg), P (1514.1 mg/ kg), S (500025.97 mg/ kg), Na (530.11 mg/ kg), B (26.99 mg/ kg),
Al (26.33 mg/ kg) and Fe (16.18 mg/ kg). Analytical values revealed nutritional
properties and mineral contents of blackthorn fruit. Moisture content, crude protein,
crude oil, crude fibre, crude energy, water soluble extract and alcohol-soluble extract
values obtained by Marakoglu et al. (2005) was 69.37%, 3.4%, 2.06%, 4.6%, 249
kcal/g, 78.28% and 14.74%, respectively. This paper aims at studying physical-
chemical characteristics and properties of blackthorn fruits growing wildly in
spontaneous flora in Oltenia region in order to identify important genotypes not only as
tree-growing but also as food additive point of view.

MATERIAL AND METHOD

There have been identified and labelled 15 genotypes of Prunus spinosa L.
growing wildly in southern part of Oltenia region, Dolj county, namely Gura Vaii village,
and Calopar village, Dolj county. Each label contained some codes such as the initial
of the village (fruit where harvested from) and the genotype number according to their
identification. Moreover, geographical coordinates were established for each type
using Altimeter programme. There were harvested 100 fruits from each genotype,
namely 7 genotypes from Gura Vaii and 8 genotypes from Calopar village, and then
were taken to the laboratory in cooled plastic bags. Next, they were labelled and
130
measured (length and diameter of the fruit, unit mass, fruit volume, fruit density, kernel
and pulp mass). After that they were prepared for chemical analyses (total dry matter,
soluble dry matter content, acidity). The length and diameter of fruit were measured by
digital micrometre. The fruit mass was measured by electronic balance. Water and
total dry matter were determined by using water evaporation method from the fine-
grained analytical average sample by maintaining the sample in a thermoregulatory
oven reaching temperatures no higher than 85-105°C (Nour 1998). The results are
expressed using millimetre for length and diameter, grams for fruit mass, water
percentage and total dry matter. Total dry matter was established using refractometric
method (Ionica 2014) using digital refractometer and the results were expressed as
percentages. The titrable acidity was determined by titration of a known amount of fruit
water extract with 0.1N NaOH using phenolphthalein as an indicator. The results are
expressed as g acid/100g fresh matter.
Statistical analysis. The data obtained after the above evaluation have been
processed so as mean values and standard deviation or standard errors. This
treatment was carried out using Data Analysis option in Microsoft Office Excel.

RESULTS AND DISCUSSION

In this study the chemical and physical properties of 15 genotypes of blackthorn
fruits growing wild in the spontaneous flora in Oltenia region were assessed. After the
investigations carried out in the laboratory, the highest average fruit weight was
registered in genotypes in Gura Vaii village 3.07± 0.47 g (GV3) whereas the lowest
amount was established in genotypes in Calopar village (2.01 ± 0.16 g la C5). The
highest fruit unit mass was observed in GV3 genotype in Gura Vaii village (4.20g)
whereas C6 genotype in Calopar was lower (3.10 g.). The minimum mass of fruits was
equal for both genotypes 1.10 g for both GV1 and C2. As regards the pulp mass, the
highest level was observed in GV1 genotype (93.75%) whereas the lowest one was
observed in C1 genotype (61.53%) (Table 2). For 4 of 7 genotypes in Gura Vaii village
the pulp mass was higher than 90% whereas only one genotype in Calopar village
reached this point. The medium length fruits for 15 genotypes was reported at the level
of 14.25 mm for genotypes in Gura Vaii village and 12.96 mm in Calopar village. The
highest rates for medium length fruits were observed in 2 genotypes from Gura Vaii (GV8
and GV3) ranging from 17.74 mm to 17.11 mm, respectively, compared to the values
obtained for the same characteristic in two genotypes of Calopar population, namely
14.04 mm (C6) and 13.89 (C5) (Table 1). The maximum height of fruits was established
as 22 mm for GV8 genotype in Gura Vaii village whereas C1 genotype in Calopar
reached only 18.80 mm. The minimum height of fruits was established as 9.50 mm for
GV 1 genotype and 9.90 mm for C3 (Table 3). The large diameter of the fruit had a
maximum value at GV8 (20.70 mm) and a minimum value for the genotype GV1 (9.40
mm), and in the Calopar population, the maximum and minimum value for the same
characteristic was 18.50 mm and 9.30 mm, encountered in the same genotype (C1). The
highest small diameter was established as 20.50 mm for GV8 genotype from Gura Vaii
area and the lowest one was established as 8.80 mm for GV1 genotype. As regards the
genotypes in Calopar area the highest and the lowest degree was established as 18.10
mm and 9.20 mm, respectively, for C1 genotype.
The average results of this study were established as being higher than those of
Marakoglu et al. (2005) (Table 1 and Table 2) for fruits growing wild in Konya province in
Turkey, except for diameter of fruit and sphericity which were almost similar to those
observed for genotypes from Calopar village namely: length (12.12 mm), unit mass (1.28
mm), fruit diameter (12.59 mm), geometric mean diameter (12.43), sphericity (1.03) and
volume (1288.5 mm³). Coefficient of variation for each value is given in table 1 and 2. For
131
C1 genotype from Calopar area and for GV1, GV2 and GV8, from Gura Vaii, the
variation coefficient for kernel mass shows greater percentages which were established
as follows: 20.00%, 27.78%, 20.83% and 25.71%, respectively, which show a great non-
uniformity for this feature. The same non-uniformity was also observed in genotypes C1,
C6, GV1, GV8; in case of kernel percentage the coefficient of variation was 20.57%,
21.49%, 24.82% and 24.57%, respectively. In terms of fruit weight, the variation
coefficient indicates a small/medium variation.
As regards physical characteristics of blackthorn fruits we can mention that the
values of mass, geometric mean diameter and sphericity were established. The
average weight of fruits that belonged to the genotypes studied here was lower, within
the range of 1.46 ± 0.19 and 3.07 ± 0.47, compared to that found by Erturk et al.
(2012) for Turkish genotypes, 5.86 g for red fruits and 7.83 g for dark-purple fruits.

Fig.1 Percentage distribution for fruit Fig.2 Percentage distribution for fruit
weight harvested in Gura Văii population weight harvested in Calopar population
Also, the technological properties such as geometric mean diameter of fruit
were established as 20.08 for GV8 and 18.46 for C1 genotype (the highest level).
Minimal values were 9.30 for GV1 genotype and 9.83 for C2 genotype, whereas the
highest average values were established as 16.84 and 16.33 for GV8 genotype and
GV3 genotype, respectively. Sphericity (shape index/form index) had average values
ranging between1.02 and 1.07 for genotypes in Gura Vaii area and from 0.98 to 1.06
for genotypes in Calopar area.
Concerning the fruit weight, of the total fruit harvested from the 7 genotypes
belonging to Gura Vaii population, only one fruit had the minimum weight of 1.10 g,
representing 0.17% of total fruit, and one single fruit had the maximum weight of 4.20
g. In the range of weight between 1.20 and 2.00 g were 43.96%, between 2.00 g and
3.00 g were 47.58%, and between 3.00 and 4.00 grams were 8.10% of the fruit (Figure
1). Of total fruit harvested from the eight genotypes belonging to Calopar population,
75.73% of fruits weighed between 1.10 and 2.00 g, 24.12% of the fruit weighed
between 2.00 and 3.00 g, and a single fruit had 3.10 g, representing 0.15% of total fruit
(Figure 2). To determine the fruit quality, chemical analyses were performed and the
results are presented in Table 3. The highest value for titratable acidity was found in
GV3 genotype (2.78 g malic acid / 100 g fresh product), and the lowest in C3 genotype
(1.21 g malic acid / 100 g fresh product). A high percentage of total dry substance
showed fruits from genotype GV8 (37.66%) and the lowest content was noted for
genotype C2 (27.71%). Considering soluble dried substance, the values ranged from
17.50% (GV5 genotype) to 25.50% (C5 genotype). Our results on titratable acidity
(Table 3) are similar to those found by Erturk et al. (2012) in Prunus spinosa L.
genotypes with dark purple, red and yellow purple fruits in Turkey; the highest acidity
was found in dark red fruits (4.99%) and the lowest in dark purple fruits (3.87%).
132
133
134
 Table nr. 3
Values and percentages of total dried substance, acidity, soluble dried substance of
Prunus spinosa L. from Gura Vaii (GV) and Calopar areas, Dolj county

No Genotype Total dry Titratable acidity Soluble dry matter

. matter % (100 g malic acid %
content/fresh fruit)
1 GV1 33.94±0.02 1.94±0.05 20.50±0.05
2 GV2 32.50±0.01 2.08±0.03 23.50±0.06
3 GV3 31.89±0.03 2.78±0.02 23.50±0.15
4 GV4 31.11±0.10 2.28±0.01 19.50±0.02
5 GV5 27.83±0.01 1.91±0.06 17.50±0.25
6 GV7 28.51±0.02 1.27±0.15 18.00±0.04
7 GV8 37.66±0.01 1.47±0.25 24.50±0.06
8 C1 36.74±0.01 1.88±0.03 23.50±0.03
9 C2 27.71±0.01 1.51±0.04 19.50±0.02
10 C3 28.12±0.10 1.21±0.02 18.50±0.35
11 C4 30.03±0.15 1.71±0.01 19.00±0.04
12 C5 35.73±0.02 1.74±0.02 25.50±0.05
13 C6 29.37±0.04 1.54±0.31 20.50±0.04
14 C7 31.21±0.06 2.11±0.25 20.50±0.05
15 C8 31.20±0.05 1.61±0.04 20.50±0.06

Marakoglu et al. (2005) report an acidity of 1.97% and a pH of 3.53% for fruit
in Konya, Turkey. The fruits of Prunus spinosa L. in Madrid, Spain, studied by Ruiz-
Rodríguez et al. (2014) showed a high content of anthocyanins (1431.75 mg
pelargonidine 3-glucoside Eq/100 g FW) and phenolic acids (728.81 mg E q gallic
acid/100 g FW), low vitamin C (11.27 mg ascorbic acid/100 g FW) and a high level of
total phenolic compounds (2294.57 mg/100 g FW). Wolbis et al. (2001) isolated from
the flowers of Prunus spinosa L. a mixture of alpha and beta amyrine, a mixture of
ursolic and oleanic acids, ursolic acid, a mixture of beta-sitosterol, gamma sitosterol 3-
0 beta-D-glucopyranoside were isolated.

CONCLUSIONS
Determining the physical and chemical characteristics of the fruits is a step
that precedes the establishment of their edible mass and an important step for the
qualitative and quantitative assessment of production and the drawing of the fruit
utilization directions. Fruit weight ranged between 1.10 g and 4.20 g and the
percentage of pulp mass ranged between 61.53 and 93.75 %. The soluble dry matter
content ranged from 17.50% to 25.50% and the titratable acidity was expressed as g of
malic acid/100 g of fresh product ranging from 1.27 to 2.78. Through this study it was
shown that there is a great variability between the genotypes of blackthorn, in terms of
characteristics of fruit under the same ecological conditions, and these genotypes
could be important as tree-growing point of view and also as food additive.

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Iliescu A.F. 2002. Cultura arborilor și arbuștilor ornamentali. Editura Ceres.
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Ionică M.E. 2014. Metode de analiză și control al calității fructelor și legumelor
proaspete și divers prelucrate. Editura Universitatea Craiova, EUC ProUniversitaria,
Craiova: 24-27.
Irizar A.C., Fernandez M.F., González A.G., Ravelo A.G. 1992. Constituents of
Prunus spinosa. Journal of Natural Products 55(4): 450-454.
Kavaz Yuksel, A. 2015. The effects of blackthorn (Prunus spinosa L.) addition on
certain quality characteristics of ice cream. Journal of Food Quality 38(6): 413-421.
Marakoglu T., Arslan D., Özcan M., Hacıseferogulları H. 2005. Proximate
composition and technological properties of fresh blackthorn (Prunus spinosa L. Subsp
dasyphylla (Schur.) fruits. Journal of Food Engineering 68(2): 137-142.
Nour V. 1998. Metode de analiză și control în industria conservelor de legume și
fructe. Editura Europa, Craiova: 28-30.
Olszewska M., Wolbis M. 2001. Flavonoids from the flowers of Prunus spinosa L.
Acta Poloniae Pharmaceutica 58(5): 367-372.
Olszewska M., Glowacki R., Bald M.W.E. 2001. Quantitative determination of
flavonoids in the flowers and leaves of Prunus spinosa L. Acta Poloniae
Pharmaceutica 58(3): 199-204.
Özcan T., Bayçu G. 2008. Fatty acid and amino acid profiles in the fruits of Prunus
spinosa L. subsp. dasyphylla (Schur) domin from Europe-in-Turkey. Advances in
Molecular Biology 1: 39-46.
Pârvu C. 2005. Enciclopedia plantelor – plante din flora României (Vol. IV). Editura
Tehnică, București.
Ruiz-Rodríguez B.M., de Ancos B., Sánchez-Moreno C., Fernández-Ruiz V., de
Cortes Sánchez-Mata M., Cámara M., Tardío J. 2014. Wild blackthorn (Prunus spinosa
L.) and hawthorn (Crataegus monogyna Jacq.) fruits as valuable sources of
antioxidants. Fruits 69(1): 61-73.
Sikora E., Bieniek M.I., Borczak B. 2013. Composition and antioxidant properties
of fresh and frozen stored blackthorn fruits (Prunus spinosa L.). Acta Scientiarum
Polonorum Technologia Alimentaria 12(4): 365-372.
Wolbis M., Olszewska M., Wesolowski W.J. 2001. Triterpenes and sterols in the
flowers and leaves of Prunus spinosa L. (Rosaceae). Acta Polonia e Pharmaceutica
58(6): 459-462.
Veličković J.M., Kostić D.A., Stojanović G.S., Mitić S.S., Mitić M.N., RanĎelović S.
S., ĐorĎević A.S. 2014. Phenolic composition, antioxidant and antimicrobial activity of
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 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
 Food produce process ing
UNIVERSITY OF CRAIO VA
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

RESEARCH CONCERNING THE BEHAVIOUR OF WALNUT GENOTYPES

WITH DIFFERENT GEOGRAPHIC ORIGIN UNDER THE ECOLOGICAL
CONDITIONS OF NORTH OF VÂLCEA COUNTY
1 1 1 1,2
Giura Simona , Scutelnicu Anca , Preda Silvia , Botu Mihai ,
3 2
Manthos Ioannis , Mitrea Rodi
1
University of Craiova – SCDP Vâlcea, Rm. Vâlcea
2
University of Craiova, Faculty of Horticulture, Department of Horticulture and Food Science, Craiova
2
University of Craiova – SCDP Vâlcea, Rm. Vâlcea
3
Institute of Plant Breeding & Genetic Resources, Dept. of Nut Trees, HAO-DEMETER , Lamia, Greece
* Correspondence author. E-mail: [email protected]

Key words: Juglans regia L., cultivars, ecological conditions, frost

ABSTRACT
The common walnut nut finds favorable conditions for cultivation in the Oltenia region
located into the SW of Romania, where the climate is temperate, with some Mediterranean
influences. Present study is evaluating during 2016 and 2017 years a total number of 26
accessions of walnuts of different geographic origins (14 from Romania, 9 from the USA, 1 from
France, 1 from Russia and 1 from Japan) from the UCv-SCDP collection regarding yield,
phenology, fruit maturation and resistance to late spring frosts.

INTRODUCTION
Common walnut (Juglans regia L.), known also as Carpathian, Persian or
English walnut is a very valuable nut crop. Although walnut is adapted to various
ecological conditions from almost 50 countries from both hemispheres, it is quite
susceptible to low temperatures during winter, but also to early and late frosts.
In Romania, the walnut culture dates back to the ancient Dacians, when the
Latin poet Ovid wrote about walnut that "he is not pretentious, he grows by the side of
the road, and he is not afraid neither of the wind, nor of rain, nor of heat or of cold"
(Cociu et al., 2008).
Walnuts can be found almost in all suitable regions of Romania from the plains
till high hills. Walnut cultivation has become very important due to the advantages of its
products (fruits, wood, leaves) (Botu et al., 2010).
Favorable ecological conditions for walnut cultivation are found in the Oltenia
region, a region located in southwestern Romania, where the climate is temperate, with
Mediterranean influences, with an average annual temperature of 10-11,5°C.
(Cosmulescu et al., 2012).
Survival and obtaining good fruit production in temperate continental areas
depends on winter temperatures (Olsen, 2010), and especially late spring frosts (Arora
and Rowland, 2011).
Juglans regia L. is a species of high plasticity and, to a certain extent, tolerates
low winter temperatures in different geographic areas and does not suffer winter
damage (Améglio T. et al., 2001).
137
 Even inside Juglans regia species there are differences regarding resistance
to low temperatures during winter, respectively to early or late frosts. Walnut cultivars
with lateral bearing proved to be more susceptible from this point of vue then the
cultivars with terminal bearing (so called Carpathian walnuts).
Resistance to winter temperatures is not just a characteristic of varieties,
according to Ameglio et al. (2001), low temperature resistance is due to the
accumulation of carbohydrates during the previous year and its storage in the trunk.
Terziev (2002) states that lesions are more severe when trees have been
exposed to low temperatures for a longer period of time.
Gandev (2013) mention that the French cultivar Lara is less winter hardy then
another French cultivar, Fernor and Bulgarian cultivar Izvor 10 regarding reproductive
organs. There are also differences between types of reproductive organs, catkins
being usually more susceptible to frost then female buds.
Germain et al. (1999) mentioned that the Californian varieties (lateral bearing
cultivars) were inappropriate for cultivation in the colder climate of France.
According to Nedev et al. (1983), low temperature damages in walnut cultures
was reported only when temperatures dropped below -20°C. The Carpathian walnut is
considered to be the most resistant, at temperatures that can reach up to -32°C; -35°C
(Domoto, 2002).
Botu et al. (2010) report that in the Vâlcea area the winter temperature of -
22.7°C did not affect the growth and fruiting of the Californian and French lateral
bearing cultivars.
Nedev et al. (1983) and Terziev (2002), cited by Gandev (2013), mention that
the temperature of -24.4°C is crucial for walnut cultivars.

MATERIAL AND METHODS

The study was conducted between 2016 and 2017 in the walnut collection
belonging to the University of Craiova - SCDP Vâlcea. The collection is located at 260
m elevation, at 45°8'45" N and 24°22'01" E, under non-irrigated conditions. Average
annual rainfall in the area is 715 mm.
The biological material used in this study consists of 26 accessions of walnut
with different geographical origins: 14 genotypes from Romania, 9 from the United
States, 1 from France, 1 from Russia and 1 from Japan. All genotypes are grafted on
Juglans regia seedling. The planting distance from the collection is 9.0 x 8.0 (139
trees/ha). The age of trees in the collection varies between 18 to 22 years old.
Research methods are related to the aims of the paper and include comments
on:
 geographic origin of studied cultivars;
 vigor of the trees;
 fruit bearing type;
 blooming time of female and male flowers;
 dichogamy;
 influence of low temperatures on vegetative buds, mixed buds and catkins;
 resistance to low temperatures during spring (March and April).
For observations on the influence of temperatures on buds, twigs were
collected from trees at a height of 2.5-3 m above the ground. 3 trees of each cultivar
were sampled, from each tree 10 twigs were harvested. The different types buds were
sectioned with a scalpel, and the percentage of frost damage was reported using a
binocular microscope.

138
 RESULTS AND DISCUSSIONS
The walnut genotypes with different geographical origins, studied in the
Northern area of Oltenia (Vâlcea county), showed differences regarding growth, fruit
bearing and fruit maturation.
In case of genotypes with geographical origin in Romania, the plant vigor
ranges from medium (Velniţa, HC O2 and HC O3) to the high (Germisara, Jupâneşti,
Sibişel 44, Valcor, Valrex, Valmit, Valcris, Unival, Valstar, Sarmis and VL 102 H).
Accessions originating from France, the USA, Japan and Russia exhibited
from high vigor (all of the genotypes from USA, except Juglans nigra, and Franquette
from France), genotypes from Japan (J. sieboldiana), Russia (J. mandshurica) and J.
nigra presented very high growth vigor (Tables 1 and 2).
The walnut accessions, which were evaluated, have three types of bearing
fruits:
- terminal: 11 genotypes from Romania (Table 3) and 7 foreign ones;
- lateral: 2 genotypes in Romania (HC O2 and HC O3) (Table 1) and 4
foreign genotypes (Table 2).
- Intermediate: Germisara cultivar from Romania and American cultivar
Hartley.
During the study period phenological data was recorded. Pollination is very
important to achieve yield. Walnut pollination is done by wind.
Walnuts are monoecious (Polito, 1996; cited by Krueger), bearing staminate
and pistillate flowers, separately located on the same tree. The walnut cultivars are
generally cross compatible. The period of pollen shedding does not completely overlap
the period of female pistillate flowers receptivity, which is known as dichogamy (Forde
and Griggs, 1975; cited by Krueger).
Dichogamy can be influenced by environmental conditions; same cultivar under
different ecological conditions can have slightly modified behaviour regarding blooming
time and dichogamy. In some exceptional climatical years, dichogamy can be modified,
a known protandrous cultivar can switch to homogamous type or a protogynous one
can pass to homogamous. Passing from protandrous to protogynous type of
dichogamy or from protogynous to protandrous type in case of same genotype under
exceptional variable climatic conditions of same location are extremely rare.
Phenology under normal climatic conditions was recorded in case of walnut
genotypes during spring of 2016.
Out of the Romanian walnut genotypes, 10 proved to behave during the study
period as protogynous and 4 as protandrous (Jupâneşti, Valcor, Valrex and Sibişel 44).
6 of the foreign accessions behaved as protogynous and 6 as protandrous (Tables 1
and 2).
In 2017, the cold wave of air of -2.1°C recorded on April 22nd has affected
drastically the flowering of walnut genotypes (Figure 4). The beginning of blooming for
Romanian genotypes occurred at the end of April, and the end of the blooming took
place beginning of May. Earlier accessions from this point of vue were HC O2, HC O3
and Valmit, and the latest one was Velniţa.
In the case of the foreign genotypes, Hartley and Serr cultivars exhibited early
blooming. J. sieboldiana, Franquette and J. nigra accessions proved to be the most
late. The blooming time began in the first 10 days of May.
st
Table 4 represents the absolute minimum temperatures for the January 1 to
st th
June 1 2017 period. The cold air reached -1.7°C on March 27 , 2017, when bud
break of walnut genotypes occurred for most genotypes (Table 3). Bud break took
place in spring of 2017 almost 3 weeks earlier than the multi-annual average period.

139
140
141
 Table 3
Bud break of the walnut cultivars studied in 2017 at UCv-SCDP Vâlcea

No. Accession Start of End of No. Accession Start of End of

name bud break bud break name bud break bud break

1 Germisara 07.04.2017 15.04.2017 14 Valmit 22.03.2017 31.03.2017

2 Jupâneşti 03.04.2017 14.04.2017 15 J. mandshurica 04.04.2017 13.04.2017
3 Franquette 27.04.2017 20.05.2017 16 J. sieboldiana 16.03.2017 31.03.2017
4 Hartley 03.04.2017 13.04.2017 17 J. nigra 05.05.2017 17.05.2017
5 Velniţa 31.03.2017 13.04.2017 18 Valcris 24.03.2017 03.04.2017
6 Adams 10 05.04.2017 13.04.2017 19 Unival 24.03.2017 03.04.2017
7 Serr 21.03.2017 31.03.2017 20 Valstar 24.03.2017 07.04.2017
8 Vina 24.03.2017 07.04.2017 21 HC O2 22.03.2017 31.03.2017
9 Sibişel 44 10.04.2017 17.04.2017 22 HC O3 20.03.2017 24.03.2017
10 Manregian 24.03.2017 03.04.2017 23 Sarmis 21.03.2017 04.04.2017
11 Pedro 05.04.2017 13.04.2017 24 Idaho 24.03.2017 08.04.2017
12 Valcor 24.03.2017 11.04.2017 25 VL 102 H 05.04.2017 16.04.2017
13 Valrex 24.03.2017 04.04.2017 26 Payne 21.03.2017 03.04.2017

Table 4
Minimum daily temperatures recorded at Rm. Vâlcea Weather Station (2017)

Climatic Year Monthly temperature values (°C)

element January February March April May June

Daily low -18.2 °C -9.3 °C -1.7 °C -2.1 °C 6.8°C 10.1°C

th th th nd th th
temperature 2017 (Jan.10 ) (Feb. 11 ) (Mar. 27 ) (Apr. 22 ) (May 11 ) (June 10 )
(°C)

After frost, twigs were harvested from each accession and samples of buds
were analyzed.
The data presented in Table 5 show that frost caused a different degree of
deterioration of the vegetative, mixed buds and catkins to the studied genotypes.
In case of Romanian genotypes the vegetative buds were affected from 0%
Germisara, Velniţa and VL 102 H) to 70.8% (Sarmis). Values over 50% also were
recorded for Valcor and HC O2 genotypes. Lower values of damage due to frost were
observed for Valrex, Valmit, Valcris and Unival cultivars.
Mixed buds were affected differently from vegetative ones. Valmit, Unival and
Jupâneşti cultivars did not show any damage, while the HC O2, HC O3 and Sarmis
genotypes emphasized 60-70% damage (Figure 1).
The male flowers disposed in drooping catkins very affected by frost. Catkins
from Velniţa, Sibişel 44, Valrex and Valcris cultivars and those from HC O2 and HC3
selections were affected in proportion of 70-100% (Fig. 2). Only 20% of the catkins
from Germisara cultivar were affected (Table 5).
The degree of damage caused to the group of American accessions varied
from 0% (Hartley, Manregian and Pedro) to 100% (Payne) in case of vegetative buds.
Also, vegetative buds of Serr, Idaho, Vina and J. nigra were affected in high
percentages (42.5 - 63.1%). Mixed buds were damaged in different proportions (100%
in case of Serr and Payne; 66.6% for Adams 10; 70% for Idaho; 60.0% for Manregian).
Catkins kept the same trend of damages (100% for Serr; 85.7% for Manregian; 71.8%
for Pedro; 69.5% for Hartley) (Figure 3).
In case of the French cultivar Franquette the vegetative and mixed buds were
only slightly affected by frost in the spring of 2017 due to their late bud break. This is in

142
accordance with Gandev and Dzuhvinov (2014) to confirm various frost susceptibility of
walnut cultivars attributed to different phenological development.
Table 5
Influence of frost on walnut accessions in the North of Vâlcea County
No. Origin Accession Mixed buds Vegetative Catkins
name affected buds affected affected
(%) (%) (%)
1 Germisara 20 0 17.9
2 Jupâneşti 0 33.3 34.3
3 Velniţa 20 0 100
4 Sibişel 44 30 44.1 70.2
5 Valcor 30 61.2 30.0
6 România Valrex 30 10.5 95.5
7 Valmit 0 20 25.6
8 Valcris 45.4 18.1 79.4
9 Unival 0 24 37.8
10 Valstar 33.3 44 26.9
11 HC O2 70 57.1 100
12 HC O3 60 45 86.9
13 Sarmis 70 70.8 66.6
14 VL 102 H 41.6 0 22.9
15 Hartley 0 0 69.5
16 Adams 10 66.6 29.1 51.3
17 Serr 100 63.1 100
18 Vina 30 42.5 29
19 U.S.A. Manregian 60 0 85.7
20 Pedro 0 0 71.8
21 Idaho 70 45.4 60
22 Payne 100 100 63.2
23 J. nigra 0 70.2 67.4
24 France Franquette 0 0 46.6
25 Japon J. sieboldiana 70 61.7 69.7
$26 Russia J. mandshurica 0 21.2 55.2

Fig. 1. Walnut mixed bud affected by frost in Fig. 2. Walnut catkin affected by frost
2017. in 2017.

143
 Fig. 3. Walnut vegetative bud affected by frost in 2017

Fig. 4. Walnut field collection from UCv-SCDP Vâlcea affected by frost in 2017

J. sieboldiana accession originating from Japan was affected by frost with

damages ranging from 61.7% in case of vegetative buds to 70.0% for mixed buds.
The mixed buds of J. mandshurica accession, originated in Russia, were not
affected by frost while 55.2% of the male flowers were destroyed.
Observation will have to be continued the next spring in order to fully evaluate
the consequences of 2017 frost on the next year yield.
Also, taking into account the visible variability of the climatic conditions, long
term phenological data collection is necessary.

144
 CONCLUSIONS
During the study period (2016 – 2017) the genotypes originating from U.S.A.,
grown under the environmental conditions from North of the Vâlcea county, were most
affected by frost occurred in April 22nd 2017.
The Franquette cultivar originating from France was the least affected by the
low temperatures in the spring of 2017 due to the later bud break and phenological
development.
Further observation should continue in the next years taking into account the
visible climatic variability.

ACKNOWLEDGMENT
Present work has been carried out with the help of Sectorial Project A.D.E.R.
3.2.2. No. 322/2015 funded by MADR.

REFERENCES
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Botu, M., Tudor, M., Botu, I., Cosmulescu, S., Papachatzis, A., 2010. Evaluation of
Walnut Cultivars in the Conditions of the Oltenia’s Hill Area Regarding Functioning
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Produselor Agricole, Ingineria Mediului. Vol. XV (XLXI). Ed. Universitaria – Craiova;
94-103.
Botu, M., Botu, I., Tudor, M. & Papachatzis, A. 2010. Advantages and disadvanta-
ges offered by growing lateral bearing walnut cultivars in the Sub-Carpathian area of
Oltenia. Scientific papers of the R.I.F.G Pitesti, XXVI, 32-35.
Cociu, V. et al. 2008. Culturile nucifere. Edit. Ceres, Bucureşti.
Cosmulescu, S. and Botu, M. 2012. Walnut biodiversity in south-western Romania
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Domoto, P. A., 2002. Walnut tree named „Domoto‟. United States Plant Patent №
US PP12,898 P2.
Gandev, S. 2013. Winter hardiness of reproductive organs of the walnut cultivars
Izvor 10, Lara and Fernor at extreme low temperatures. Bulgarian Journal of
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Gandev, S., Arnaudov, V., Dzhuvinov, V., Koumanov, K. and Perifanova -
Nemska,
M. (2013). Agrobiological evaluation of 'Lara' walnut cultivar under the climatic
conditions of southern Bulgaria. Acta Hortic. 981, 141-146
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Gandev, S. and Dzhuvinov,V. 2014. Evaluation of Some Walnut Cultivars under
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Germain E., Prunet Y-P. and Garcin A. 1999. Le Noyer. CFIFL, Paris, PP-93-95.
Krueger, W.H. 2010. Pollination of English Walnuts: Practices and Problems.
HortTehnology 10(1). Pollination Workshop Proceedings.
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Nedev, N., Serafimov, S., Anadoliev, G., Kavardjikov, L., Krinkov, H., Radev, R.,
Dochev, D., Stamatov, I., Slavov, N., Vishanska, Y., Rusalimov, J., Iovchev, I.,
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Djeneva, A., Lalev, N., Iliev, I., Slavcheva, R. 1983. Fruit Nuts. H. G. Danov, Plovdiv,
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Olsen J. E. 2010. Light and temperature sensing and signaling in induction of bud
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 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
UNIVERSITY OF CRAIO VA  Food produce process ing
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

COMPARATIVE STUDY ON POLYPHENOLS CONTENT AND

ANTIOXIDANT EFFECT OF OLIVE CULTIVARS FROM THE THESSALY
REGION, GREECE
1 2 3
Gougoulias Nikolaos *, Kamvoukou Christina- Anna , Ntalla Maria-Nektaria ,
1
Papachatzis Alexandros
1
Department of Agricultural Technology, Technological Educational Institute of Thessaly, 41110 Larissa,
Greece
2
School of Pharmacy, Aristotle University of Thessaloniki, Greece
3
Department of Computer Science and Engineering, Data Analysis, Technological Educational Institute of
Thessaly, 41110 Larissa, Greece
*Correspondence author. email: [email protected]

Key words: Antioxidant activity FRAP; Antiradical activity DPPH; Olive fruits; Total phenols

ABSTRACT
Seven different varieties olive fruits (Amfisis, Chalkidikis, Kalamon, Megaritiki,
Koroneiki, Arbequine and Arbosana) cultivated on the region of Larissa, Greece were studied for
total polyphenol content, phenolic fractions and for antioxidant properties. It has been
established that total phenols content, move in broad ranges depending on the variety, chemical
soil properties, and with the ecological conditions. The content of TP in the olives varieties varied
from 9.48 to 19.63 mg (GAE)/g FW, (NFP) from 1.43 to 5.11 mg (GAE)/g FW and (FP) from
5.89 to 14.52 mg (GAE)/g FW. The antioxidantactivity FRAP of the seven varieties of fresh fruits
ranges from 37.2 to 77.7 μmol (AAE)/g FW. The antiradical activity (DPPH•) ranges from 158.41
to 281.31 µmol (Trolox)/g FW. The olives grown on the region of Larisa, Greece offer a high
intake of antioxidants for the prevention of many diseases.

INTRODUCTION
The fruits are natural reservoir of phenolic compounds and of natural
antioxidants (Gougoulias 2014). Olive fruits (Olea europaea L.) are one traditional
Greek product, component of the Mediterranean diet, and natural reservoir of phenolic
compounds and of natural antioxidants (Boskou & Visioli 2003). Olive fruits are rich to
flavonoids, secoiridoids, phenolic acids and phenolic alcohols, which exert high
antioxidant activity (Dağdelen et al. 2013). Phenolic compounds influence the sensory
properties of the olive fruits (Brenes et al. 1995).
The concentration and composition of phenolic compounds in the olive fruits
depends from the cultivar, the irrigation and the climatic conditions (Romani et al.
1999, Tovar et al. 2002, Vinha et al. 2005).
The aim of the present study is to determine the content of phenolic
compounds and the antioxidant activity, of olive fruits from different varieties, grown on
the Thessaly region, Greece.

147
 MATERIAL AND METHODS
Experimental: The study was carried in the farms Alatas, during the 2015 -
2016 seasons, to two olive orchards. The one olive orchard (Α) is located on the region
Gonnoi of Larissa (latitude 39°51′N, longitude 22°29′E, 200m altitude), where are
grown the olive varieties Amfisis, Kalamon, Chalkidikis and Megaritiki. The olive trees
are over a hundred years old, with distance between two consecutive trees 6m and
3
between two consecutive lines 7m. 2000 m irrigation water per hectare were added.
Also, 50 kg sheep manure corresponding to 1.20 kg Nitrogen, 1.10 kg Potassium, 0.4
kg Phosphorus and 13.8 kg Organic matter per tree were added. The other olive
orchard (B) is located on the region Kranona of Larissa (latitude 39°26′6″N, longitude
22°17′55″E, 100m altitude), where are grown the olive varieties Koroneiki, Arbequine
and Arbosana. The olive trees are fifteen years old, with distance between two
3
consecutive trees 3m and between two consecutive lines 4m. 4500 m irrigation water
per hectare were added. Also, 30 kg sheep manure corresponding to 0.65 kg Nitrogen,
0.60 kg Potassium, 0.2 kg Phosphorus and 7.8 kg Organic matter per tree were added.
The climate in the area is continental, with cold rainy winters, average temperatures
0 0
6 C, hot summers, average temperatures 25 C and average annual precipitation 420
mm.
The basic characteristics of olive fruits are shown in Table 1. The collection of
fruits took place at November of 2016, at the stage of complete maturation. Two trees
by each olive variety were chosen, were sampled of olive fruits one kilogram with four
replicates from each tree, from all the orientations and without type of disease. These
samples were stored at -18oC and were further subjected to analyses.
.
Table 1
Basic characteristics of olives

Varieties Size Color Use Origin

Amfisis Big Black Dual-purpose Greece
Chalkidikis Big Yellowish-green Table olives Greece
Kalamon Big Black Table olives Greece
Megaritiki Medium Black Dual-purpose Greece
Koroneiki Small Black Oil production Greece
Arbequine Small Black Oil production Spain
Arbosana Small Black Oil production Spain

Preparation of the methanol extracts: The flesh was separated from the
kernel and 100 g of flesh for each treatment were subjected to freeze drying for further
extraction and determination of humidity. The dry mass was crushed and stored in
clean bottles in refrigeration (Boskou et al. 2006). Two g of dry sample was extracted
two times with 50 mL of 80% aqueous methanol for 24 h at 150 rpm, the methanolic
extracts were combined and washed two times with 25 mL n-hexane in order to
eliminate the oil of the methanolic extract (Rigane et al. 2011). The separation of the
phases was performed with separating funnels. Subsequently the methanolic extract
was evaporated under nitrogen, and the residue was dissolved in 50 mL of 80%
aqueous methanol, stored in clean bottles in refrigeration in the dark until its use.
Methods of analyses: Total polyphenolic (TP) content was determined with
the Folin-Ciocalteu (F.-C.) reagent according to the method by (Singleton & Rossi
1965) and the results were expressed as gallic acid equivalent (GAE) in mg/g fresh
weight. The content of Nonflavonoid phenols (NFP) was determined with the F.-C.

148
reagent after removing the flavonoid phenols (FP) with formaldehyde according to the
method by (Kramling & Singleton 1969) and was expressed as gallic acid equivalent
(GAE) in mg/g fresh weight. Flavanoid phenols (FP) were determined as a difference
between the content of total phenols (TP) and nonflavonoid phenols (NFP). Their
amount was evaluated as gallic acid equivalent in mg/g fresh weight.
The antioxidant activity of ferric reducing antioxidant power (FRAP) of the
methanol extracts was determined on the basis of the method by (Benzie & Strain
1999) and was expressed as ascorbic acid equivalent (AAE) in µmol/g fresh weight.
•
The antiradical activity (DPPH ) of the methanol extracts was determined according to
the method by (Brand-Williams et al. 1995) using the stable free radical 2,2′-diphenyl-
1-picrylhydrazyl (DPPH•). The activity was evaluated as Trolox equivalent (TEAC) in
μmol/g fresh weight.
Soil was analyzed using the following methods which are referred by Page
(1982).
Organic matter was analyzed by chemical oxidation with 1 mol/L K2Cr2O7 and
titration of the remaining reagent with 0.5 mol/L FeSO4.
Inorganic nitrogen was extracted with 0.5 mol/L CaCl2 and estimated by
distillation in the presence of MgO and Devarda's alloy, respectively.
Available P forms (Olsen P) was extracted with 0.5 mol/L NaHCO3 and
measured by spectroscopy. Exchangeable form of potassium was extracted with 1
mol/L CH3COONH4 and measured by flame Photometer (Essex, UK). pH and (EC),
Electrical conductivity measured in the extract (1 part soil : 5 parts H2O).
Available forms of Mn, Zn, and Cu were extracted with DTPA (diethylene
triamine pentaacetic acid 0.005 mol/L + CaCl2 0.01 mol/L + triethanolamine 0.1 mol/L)
and measured by atomic absorption. The samples were analyzed by Atomic
Absorption (Spectroscopy Varian Spectra AA 10 plus, Victoria, Australia), with the use
of flame and air-acetylene mixture (Varian 1989).
Statistical analysis: Data were analyzed using the MINITAB (Ryan et al.
2005) statistical package. The experiment had four replications. Analysis of variance
was used to assess treatment effects. Mean separation was made using Tukey‟s test
when significant differences (P=0.05) between treatments were found.

RESULTS AND DISCUSSIONS

Soil samples collected from olive orchards were analyzed for nutrient element
and organic matter content (Table 2). In the olive orchard (A) the soil was Sandy Clay
Loam (SCL) and in the olive orchard (B) Sandy Loam (SL).
In the olive fruits studied, the Kalamon variety characterised by the highest TP
content, with 19.63 mg (GAE)/g FW, followed by those of the Koroneiki and Chalkidikis
with 15.79 and 15.50 mg (GAE)/g FW respectively, and the lowest by the Megaritiki
and Amfisis with 9.64 and 9.48 mg(GAE)/g FW respectively (Table 3). Our results are
in agreement with the data obtained by other authors who have established differences
in phenols content in the different olive fruits varieties (Boskou et al. 2006). The
established trend in the variation of phenols within one and the same variety under the
action of climatic factors is observed also by other authors (Petridis et al. 2012).
The content of flavonoid phenols (FP) in the studied fruits ranges from 5.89 to
14.52 mg (GAE)/g FW (Table 3). The FP content in the fruits Amfisis, Arbosana
Arbequine, Kalamon and Koroneiki constitute 84.9%, 80.1%, 77.3%, 73.9 and 72.3
respectively of TP amount, whereas for fruits Chalkidikis and Megaritiki reached 70.3
% and 61.2%, respectively. The content of non-flavonoid phenols (NFP) ranges from
1.43 to 5.11 mg (GAE)/g FW (Table 3). The NFP content in the fruits Megaritiki,
Chalkidikis, Koroneiki, Kalamon and Arbequine constitute 38.9%, 29.7%, 27.7%,
149
26.0% and 22.7% respectively of TP amount, whereas for fruits Arbosana and Amfisis
reached 19.9 % and 15.1%, respectively.

Table 2
Soil chemical properties of olive orchards studied

Soil properties Olive orchard (A) Olive orchard (A)

Soil depth
(0-30) cm (30-60) cm (0-30) cm (30-60) cm
Texture SCL SCL SL SL
pH (1:5) 6.70 ± 0.34 6.97 ± 0.35 6.30 ± 0.30 6.72 ± 0.32
EC (1:5), dS/m 0.19 ± 0.01 0.19 ± 0.01 0.17 ± 0.01 0.15 ± 0.01
CaCO3 (%) 2.20 ± 0.11 4.89 ± 0.25 1.50 ± 0.08 1.82 ± 0.09
Organic matter (%) 2.62 ± 0.15 1.90 ± 0.10 2.00 ± 0.12 1.24 ± 0.05
Ν-inorganic (mg/kg) 192.2 ± 9.5 144.6 ± 8.3 161.4 ± 9.3 112.2 ± 7.7
K-exchangeable (mg/kg) 189.4 ± 9.1 98.6 ± 4.93 222.4 ± 9.9 164.8 ± 7.6
P -Olsen (mg/kg) 25.6 ± 4.56 13.4 ± 0.74 16.4 ± 0.97 9.3 ± 0.56
Cu-DTPA (mg/kg) 4.4 ± 0.23 2.1 ± 0.11 6.2 ± 0.28 4.2 ± 0.20
Zn-DTPA (mg/kg) 2.0 ± 0.08 0.7 ± 0.04 1.1 ± 0.06 0.4 ± 0.02
Mn-DTPA(mg/kg) 4.8 ± 0.23 2.6 ± 0.16 7.3 ± 0.45 6.2 ± 0.37
Data represent average and SE deviation, (n) = 4.

Table 3
Total phenols content, flavonoid and non-flavonoid phenols in the olives studied

Olive fruits Total phenols Flavonoid phenols Non-flavonoid

cultivars (TP) (FP) phenols (NFP)
mg (GAE)/g FW
Amfisis 9.48e 8.05c 1.43e
Chalkidikis 15.50b 10.90b 4.60b
Kalamon 19.63a 14.52a 5.11a
Megaritiki 9.64e 5.89d 3.75c
Koroneiki 15.79b 11.42b 4.37b
Arbequine 10.97d 8.48c 2.49d
Arbosana 13.09c 10.49b 2.60d
Columns with the same letter do not differ significantly according to the Tukey‟s test
(P=0.05).

The results of the FRAP antioxidative assay, of the seven varieties of fresh
fruits reveal that the Amfisis and Arbosana varieties exert the highest activity (as
ascorbic acid equivalent) with 77.7 and 74.8 μmol (AAE)/g FW respectively and those
of the Megaritiki variety the lowest with 37.2 μmol (AAE)/g FW (Table 4). These values
are higher than those reported by other authors for the same varieties, but from
different regions of Greece (Petridis et al. 2012). The correlation between the
antioxidant activity (FRAP) of fruits and the contents of ТР was low, with correlation
coefficient (r2) equal to: 0.006.
The antiradical activity (DPPH•) of fruits from the olives varieties studied
ranges from 158.41 to 281.31 µmol (Trolox)/g FW (Table 4). The Amfisis and Arbosana
varieties exert the highest activity with 281.32 and 253.12 μmol (Trolox)/g FW
150
respectively, and the Megaritiki variety exert the lowest with 158.41 μmol (Trolox)/g
FW. These values are higher than those reported by other authors for some Turkey
varieties (Gurel et al. 2014). The correlation between the antiradical activity (DPPH•) of
fruits and the contents of ТР was low, with correlation coefficient (r2) equal to: 0.08.

Table 4
Antioxidant activity FRAP and antiradical activity DPPH• in the olives studied

Antioxidant activity FRAP Antiradical activity DPPH•

μmol (AAE)/g FW µmol (Trolox)/g FW
Amfisis 77.7a 281.32a
Chalkidikis 50.2b 222.14b
Kalamon 61.8b 261.32a
Megaritiki 37.2c 158.41c
Koroneiki 51.1b 229.21b
Arbequine 55.7b 227.38b
Arbosana 74.8a 253.12a
Columns with the same letter do not differ significantly according to the Tukey‟s test
(P=0.05).
CONCLUSIONS
Phenolic compounds, antioxidant and antiradical activity of olives were studied
depends strongly on the variety and by the climatic conditions of the each region.
Olives which are cultivated on the region of Larissa, Greece are a source of bioactive
components that could be included in functional foods composition.

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Dağdelen A., Tümen G., Özcan M.M., Dündar E., 2013, Phenolics profiles of
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 Gurel S., Sahan Y., Basar H., 2014, Antioxidative properties of olive fruits (Olea
europaea L.) from Gemlik variety and relationship with soil properties and mineral
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wines. American Journal of Enology and Viticulture, 20(2), 86-92.
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Petridis A., Therios I., Samouris G., 2012, Genotypic variation of total phenol
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content in five tuscany cultivars of Olea europaea L. Journal of Agricultural and Food
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152
 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
 Food produce process ing
UNIVERSITY OF CRAIO VA
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

THE EFFECT OF POSIDONIA OCEANICA ON SOIL ORGANIC MATTER

BIODEGRADATION AND OTHER SOIL CHEMICAL PROPERTIES

Gougoulias Nikolaos1*, Goula Ioanna-Maria1, Papachatzis Achilleas3, Ntalla Maria-

2 1
Nektaria , Efthimiou Spyridoula
1
Department of Agronomy Technology, Technological Educational Institute of Thessaly, 41110 Larissa,
Greece
2
Department of Computer Science and Engineering, Data Analysis, Technological Educational Institute of
Thessaly, 41110 Larissa, Greece
3
School of Agriculture, Aristotle University of Thessaloniki, Greece
*Correspondence author. email: [email protected]

Key words: posidonia oceanica; soil chemical properties; soil organic matter biodegradation

ABSTRACT
The effects of posidonia oceanica on soil organic matter biodegradation and on soil
chemical properties an incubation experiment were studied. The air dried posidonia plant tissues
incorporated at five different rates (0, 0.3, 0.6, 0.9 and 1.2 g per 50 g of soil) resulted in
increases in organic carbon mineralization, mineral nitrogen forms, available potassium in
organic and available phosphorus. The level of available forms of Fe and Zn was increased at all
the rates of added posidonia, whereas Mn was increased at the three upper rates. The level of
available Cu did not show statistically significant differences in comparison with the control. Also,
the addition of posidonia resulted in a increase in total forms of Na, K, P and Zn. The results of
this study indicated that posidonia could be applied to the soil without any extremely negative
effect on the soil chemical and biological properties.

INTRODUCTION
The addition of organic materials in the soil, affects the composition and
biological activity of microflora. When applying of organic fertilizers in the soil,
dominate microbial nutrition (Riber et al. 2014). In biological agriculture is mandatory to
replace chemicals with natural additives. Between them, the plant debris for their role
in the soil fertility, the plant essential oils for their role as pesticides against on the soil
pathogens (Gravanis et al. 2004).
Posidonia oceanica is a Mediterranean seagrass that produces onshore
deposits. Many authors have studied the use of Posidonia oceanica as substitutes to
peat and as a growing medium for the soilless cultivation of Tomato (Mininni et al.
2015, Montesano et al. 2013). Studies have shown that the Posidonia oceanica, can
be used as composting material with poultry droppings for improving soil fertility for
horticulture (Tangour et al. 2015). Also, posidonia residues can be used as organic
mulch and soil amendment (Grassi et al. 2015).
Our previous studies have shown, the effect of oregano and basil on
degradation of organic fertilizers, and it was found that the presence of foliar tissues of
oregano, act a slowing effect on degradation of organic fertilizer, while the presence of
basil, accelerate it (Chouliaras et al. 2007, Gougoulias et al. 2010).
153
 The objectives of this research were to study, in the laboratory, if the
application of Posidonia oceanica dry matter into soil has an effect on soil organic
matter and on soil chemical properties.

MATERIAL AND METHODS

In coastal areas of the island Skopelos accumulate large quantities of
seaweed-sea Posidonia oceanica, and because of the environmental problems created
transported to landfills. The October 2015 held the collection of Posidonia oceanica
from the coastal areas of island Skopelos, Greece. Because of the high salinity,
Posidonia oceanica (EC =16.85 dS/m) stirred the material for one hour with distilled
water at ratio (1: 5), filtered, air dried, well milled and were used.
Incubation experiment: In this study, 12.3 g of a sheep manure containing 5
g of organic matter (Table 1), obtained from the farming establishments of TEI of
Thessaly, and was added to 50 g of air-dried soil that was poor in organic matter,
derived from the same region (Table 1). Into 50 g of this soil plus 12.3 g of the sheep
manure, 0, 0.3, 0.6, 0.9 and 1.2 g of air-dried and well milled of leaf tissues Posidonia
oceanica were added. Thus an experimental unit is constituted by 50 g of soil, 12.3 g
of manure and a variable amount of Posidonia oceanica. In the incubator, the
treatments were prepared in four replicates and kept at 28 0C for a period of 15 weeks.
During the first three weeks of the incubation period, the moisture was maintained at
two-thirds of field capacity, but for the next three weeks the soils were left to dry. This
process was repeated until the end of the incubation period according to (Wu and
Brookes 2005) they reported that the alternation of drying and rewetting soil samples
enhances mineralization of both soil biomass organic matter and non-biomass organic
matter. Soil aeration and regulation of moisture enhance the growth and metabolic
activity of aerobic soil microorganism (Gordon et al. 2008). At the end of the incubation
period, soil samples were analyzed.
Methods of analyses: Samples were analyzed using the following methods
which are referred by (Page et al. 1982).
Organic matter was analyzed by chemical oxidation with 1 mol/l K2Cr2O7 and
titration of the remaining reagent with 0.5 mol/l FeSO4.
Both ammonium and nitrate nitrogen were extracted with 0.5 mol/l CaCl2 and
estimated by distillation in the presence of MgO and Devarda's alloy, respectively.
Available P forms (Olsen P) was extracted with 0.5 mol/l NaHCO3 and measured by
spectroscopy. Exchangeable forms of potassium and sodium ware extracted with 1
mol/l CH3COONH4 and measured by flame Photometer. Available forms of Mn, Zn,
and Cu were extracted with DTPA (diethylene triamine pentaacetic acid 0.005 mol/l +
CaCl2 0.01 mol/l + triethanolamine 0.1 mol/l) and measured by atomic absorption.
Organic phosphorus was measured after mineralization by combustion of the
sample and subtraction of the mineral phosphorus amounts, which had previously
been estimated in the laboratory. The mineral amounts were extracted with 1 mol/l
H2SO4 and all forms were measured by spectroscopy.
For the determination of total metals Mn, Cu and Zn , 1 g of material,
digestion at 350 0C + 10 ml HNO3 + 5 ml HCLO4. According to the method described
by (Varian 1989), the samples were analyzed by Atomic Absorption (Spectroscopy
Varian Spectra AA 10 plus), with the use of flame and air-acetylene mixture.
Statistical analysis: Data analysis was made using the MINITAB (Ryan et al.
2005) statistical package. Analysis of variance was used to assess treatments effect.
Mean separation was made using Tukey's test when significant differences (P=0.05)
between treatments were found.

154
 Table 1
Chemical properties of soil samples, manure and Posidonia oceanica foliar tissues
used in the experiment

Property Soil *Manure Posidonia oceanica

Air dried material
Texture Sandy Loam - -
pH 7.74 ± 0.3 8.55 ± 0.4 7.84 ± 0.3
EC, dS/m 0.42 ± 0.02 2.93 ± 0.2 2.56 ± 0.3
Organic matter (%) 0.85 ± 0.04 40.54 ± 2.4 -
CaCO3 (%) 8.48 ± 0.6 - -
N -Total (g/kg) 1.55 ± 0.13 11.57 ± 0.8 4.55 ± 0.4
N-NH4+ (mg/kg) 28.6 ± 4.3 - -
N-NO3- (mg/kg-1) 94.1 ± 10.4 - -
K-exchangeable (mg/kg) 264.8 ± 9.2 - -
K-Total (g/kg) 5.20 ± 0.2 19.88 ± 1.3 0.71 ± 0.05
Na-exchangeable (mg/kg) 211.1 ±6.5 - -
Na-Total (g/kg) 0.61 ± 0.02 8.40 ± 0.5 10.67 ± 0.5
CEC (cmol/kg) 18.5 ± 0.9 - -
P -Olsen (mg/kg) 11.1 ± 4.2 - -
P -Total (g/kg) 0.36 ± 0.02 6.85 ± 0.2 0.48 ± 0.04
Cu –DTPA (mg/kg) 1.13 ± 0.07 - -
Zn -DTPA (mg/kg) 1.01 ± 0.06 - -
Mn -DTPA (mg/kg) 2.74 ± 0.17 - -
Cu –Total (mg/kg) - 39.6 ± 9.7 44.8 ± 3.3
Zn -Total (mg/kg) - 212.5 ± 9.8 61.6 ± 3.7
Mn -Total (mg/kg) - 77.8 ± 4.8 66.1 ± 7.1
Fe-Total (g/kg) - 5.37± 0.9 0.33 ± 0.01
*Digested sheep manure four months; Electrical conductivity, (EC) and soil pH is determined in
(1:5) soil/water extract; Data represent average means and SE deviation. (n)=4.

RESULTS AND DISCUSSIONS

The results of the laboratory experiment showed that decomposition of soil
organic matter added as manure or preexisted organic matter in soil, was increased
with the increase of posidonia concentration in soil, as compared with the control (soil
plus manure), (Figure 1). In particular, the data showed an increased mineralization of
soil organic carbon by 23.8 to 42.6 % at all rates addition of posidonia compared to the
control.
The addition of dry plant tissues posidonia in the mixture (soil plus manure), in
all treatments, in comparison with the control, at the end of the incubation period
increased available forms of P, Zn, Fe, ammonium and nitrate nitrogen (Table 2). In
contrast, available form of Cu by the addition of posidonia in the mixture, in all
treatments, at the end of the incubation period did not show statistically significant
differences in comparison with the control (Table 2).

155
Figure 1. Effect of dry posidonia added rates on soil organic C mineralization. Columns
in each characteristic of the graph with the same letter do not differ significantly
according to the Tukeyʼs test (P=0.05). C, control (soil plus manure); CP1, CP2, CP3
or CP4, control and posidonia 0.3, 0.6, 0.9 or 1.2g, respectively.

Table 2
Effect of posidonia dry matter concentration on soil available forms of elements

Treatments Available forms DTPA

P-Olsen N-NH4+ N-NO3- Cu Zn Mn Fe
(mg/kg soil)
C 276.9b 24.5b 161b 1.02a 2.38c 2.02b 12.22b
CP1 419.7a 178.8a 322a 0.92a 5.34b 2.22b 16.74a
CP2 423.5a 149.8a 357a 0.92a 5.88b 3.59a 16.42a
CP3 406.3a 145.5a 322a 0.73a 7.67a 3.17a 17.67a
CP4 422.2a 164.2a 301a 0.89a 6.74a 3.05a 17.21a
For each chemical property of soil mixtures, columns of table with the same letter do not differ
significantly according to the Tukey‟s test (P=0.05). C, control (soil plus manure); CP1, CP2,
CP3 or CP4, control and posidonia 0.3, 0.6, 0.9 or 1.2g, respectively.

The analysis of variance performed for the organic phosphorus and

exchangeable forms of potassium data indicated that the organic phosphorus and
exchangeable potassium was significantly increased by the added posidonia in all
treatments (Table 3).
Table 3
Effect of posidonia dry matter concentration on soil chemical properties

Treatments Exchangeable forms Organic EC pH

Na K P Extract (1: 5) in water
(mg/kg) (dS/m)
C 1110.9c 2130.6b 389.6c 1.66e 7.62a
CP1 2196.5b 5927.9a 517.9b 2.18d 7.55a
CP2 2334.5a 6180.1a 701.0a 2.68c 7.78a
CP3 2472.5a 5675.6a 706.9a 3.30b 7.69a
CP4 2677.2a 5675.6a 474.5b 4.12a 7.71a
For each chemical property of soil mixtures, columns of table with the same letter do not differ
significantly according to the Tukey‟s test (P=0.05). C, control (soil and manure); CP1, CP2, CP3
or CP4, control and posidonia 0.3, 0.6, 0.9 or 1.2 g, respectively.

156
 Also, the exchangeable form of sodium was increased by the added posidonia
dry matter, in all treatments. In particular, the exchangeable form of sodium was
increased with increasing posidonia dry matter concentration. Electrical conductivity
also increased in all treatments by the added posidonia. However, the greatest
increase was caused by the highest rate of dry plant tissues posidonia. The soil pH did
not show statistically significant differences compared to control, in all cases by the
added posidonia (Table 3).
The addition of dry plant tissues posidonia in the mixture (soil plus manure), at
the end of the incubation period in comparison with the control, increased total forms of
Na, K, P, Cu and Zn, in all treatments (Table 4). In contrast, total forms of Mn and Fe
by the addition of plant tissues posidonia did not show statistically significant
differences in comparison with the control, in all treatments (Table 4).

Table 4
Effect of posidonia dry matter concentration on soil total forms of elements

Treatments Total forms

Cu Zn Mn Na K P Fe
(mg/kg soil) (g/kg soil)
C 48.2a 57.6b 504.9a 1.315c 8.098c 1.454b 3.286a
CP1 26.83b 71.0a 556.9a 3.136b 10.479a 2.076a 3.347a
CP2 23.56b 85.9a 540.4a 3.136b 10.479a 2.276a 3.322a
CP3 22.91b 82.1a 524.0a 3.135b 9.431b 2.182a 3.289a
CP4 24.22b 76.8a 544.9a 4.182a 8.288c 2.048a 3.322a
For each chemical property of soil mixtures, columns of table with the same letter do not differ
significantly according to the Tukey‟s test (P=0.05). C, control (soil plus manure); CP1, CP2,
CP3 or CP4, control and posidonia 0.3, 0.6, 0.9 or 1.2 g, respectively.

CONCLUSIONS
Four different rates of posidonia biomass were applied into a soil amended
with manure, and after an Incubation experiment the effects on the soil chemical
properties were attested. These results, confirm the role of dry posidonia biomass as it
is a valuable material for soil amendment, could be applied to the soil as an
acceleration agent for soil organic matter biodegradation, improves soil chemical and
biological properties, while that recycling constitutes a useful practice for environment
protection.

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Montesano F.F., Parente A., Grassi F., Santamaria P., 2013, Posidonia-Based
Compost as a Growing Medium for the Soilless Cultivation of Tomato. In International
Symposium on Growing Media and Soilless Cultivation, 1034, 277-282.
Page A.L, Miller R.H., Keeney D.R.,1982, Methods of Soil Analysis. Part 2,
Chemical and Microbiological properties. Agronomy (9), ASSSA, Mad. Wisc. USA.
Riber L., Poulsen P.H., Al-Soud W.A., Hansen L.B.S., Bergmark L., Brejnrod A.,
Sørensen S.J., 2014, Exploring the immediate and long-term impact on bacterial
communities in soil amended with animal and urban organic waste fertilizers using
pyrosequencing and screening for horizontal transfer of antibiotic resistance. FEMS
microbiology ecology, 90(1), 206-224.
Ryan B.F., Joiner B.L., Cryer J.D., 2005, MINITAB Handbook: Updated for
release 14, 5th edition. Brooks/Cole-Thomson Learning Inc., Kentaky, KY.
Tangour D., Yeddes N., Ayadi M.T., Jrad A., 2015, Composting of Posidonia
oceanica (Neptune grass) mixed with poultry droppings residues for agricultural and
environmental purpose. International Journal of Environment and Waste Management,
15(4), 377-385.
Varian M., 1989, Flama Atomic Absorption Spectroscopy. Analytical Methods.
Varian Australia. Publ. N0: 85-100009-00.
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soil. Soil Biology & Biochemistry, 37, 507-515.

158
 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
 Food produce process ing
UNIVERSITY OF CRAIO VA
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

RESEARCHES CONCERNING THE USE OF TWO IRRIGATION SYSTEMS

WITH REDUCED CONSUMATION OF WATER IN ORDER TO CONTROL
THE WATER STRESS AT GRAPEVINE
1*
Ion Marian , Burlacu Cristian, Pircalabu Liliana, Filip Vlad Andrei, Brinduse Elena
1*
Research Institute for Viticulture and Enology Valea Calugareasca
* Correspondence author. E-mail: [email protected]

Key words: collector pluvial, humidity sensor, water stress, grapevine

ABSTRACT
The work has been carried out during 2016 – 2017 period, in a vineyard located in
Valea Călugarească viticultural center, planted with Tămâioasă romanească variety, aiming to
determine the influence of two irrigation systems (with pluvial collectors located between the
vines supplemented with a system based on dripping and irrigation system based only on
dripping) on the biological process at the soil level and on the physiological processes in the
plant, on the quality of the grape harvest. The obtained results highlight the positive effect of the
two irrigation systems on the microbiological activity in the soil. The positive effects on plants is
referring to the stimulation of the physiological processes in the plant, ensuring an optimal
growth and yielding ratio, increasing the grape production by 17-20 % and providing a sugar
content of the grape juice that allows the obtaining of wines with good quality.

INTRODUCTION
In the current context of climate change, which led to the appearance in the
vegetation period of the vine of some periods (longer or shorter) of water-stressed,
irrigation of grapevines become a necessity, even in wine-growing areas, which, two
decades ago were considered wet. Although the vine is adaptable to drought
conditions (possessing a root system well developed in depth and with a high capacity
for absorption) and the soil moisture to values close to the coefficient of wilting can be
used, the absence of water in the soil can induce critical period in terms of
physiological point of view, which can be reflected not only on the production of that
year, but also on the future evolution of crops.
Considering the high water consumption required for irrigation of the vineyard
during excessively dry, become a necessity the use of new methods for irrigation of the
vine in order to ensure a coefficient of utilization of irrigation water higher than that
obtained by using the conventional methods and which do not influence negatively the
quality of grape production (Pircalabu,2004; Ojeda, 2014). In this sense, the
researches carried out at ICDVV Valea Calugareasca within a demonstrative polygon
with the Tamaioasa romaneasca variety aimed to establish the influence of two
irrigation systems (with pluvial collectors located between the vineyards+drip irrigation

159
system and drip irrigation) on soil biological and physiological processes of plant
growth and on the grape quality.

MATERIAL AND METHODS

In the year 2016 an experimental plot was arranged in a plantation, three years
aged, with Tamaioasa Romaneasca variety grafted on rootstock SO4-4. This variety is
recognized for quality aromatic wines, recommended for Valea Calugareasca
viticultural center.
The experience was monofactorial, the factor studied being the irrigation
system with graduations: a1 = drip irrigation and pluvial collector; a2 = drip irrigation and
a3 = non-irrigated. The soil in the experimental polygon is reddish preluvosol which is
part of the luvisol class.
The electricity for the functioning of the irrigation system with low water
consumption has been assured by a solar generation of electricity.
The pluvial collectors have been placed between the vines, and were
integrated into the automated drip irrigation system.
The collectors includes a storage vessel provided with a cover, which
decreases in the rate of evaporation of water accumulated.
The geometry of the cover ensures efficient capture of rainwater that is slowly
ceded to the vine through cotton wicks.
The monitoring of the humidity threshold value at the level of the vine roots
was done by means of 8 sensors, conceived by INCDMTM Bucharest, evenly
distributed on the experimental plot, and by 3 humidity transducers.
Irrigation was started on the August 2nd when soil humidity reached the ceiling
of 17%, considered as the minimum threshold from which it must be triggered and was
stopped on August 26, although the soil moisture level fell below the minimum
threshold, given that the grapes were in the water in the veraison.
The following parameters were used for the characterization of the ecoclimatic
conditions during the experimental period: average annual temperature, sum of the
active temperature, sum of the useful temperature grades, the sum of the monthly
precipitation and the number of days with precipitation.
Observations and determinations of the main physical properties (apparent
density, hygroscopicity coefficient, coefficient of wilting, field capacity, active humidity
range) and microbiological observations (determining the viable germ load,
determining the structure of microbial communities) were made.
Determinations were made on the main physiological processes in plant
(photosynthetic rate, breathing, sweating), foliar surface index (LAI) by using a LCA-4
automated analyzer.
The grape production (Kg/vine) was determined by weighing the grapes
individually harvested in comparison with the control.
The mechanical composition of the grapes was determined by calculating the
grain technological indices (grape structure index, grain index and yield index).
The composition of the grape juice has been evaluated on the basis of the
following parameters: sugar (g / l) and total acidity.
Determinations have been made on the hydric regime of the grapevine: leaf
water potential and stomatal conductance.

160
 RESULTS AND DISCUSSIONS
The ecoclimatic conditions specific to 2017 year were characterized by a high
heliothermic regime of rich water resources in July and relatively low in August (Table
1).
In the table 2 are shown the main physical and hydro physical characteristics
of the soil in the experimental plantation.

Table 1
The ecoclimatic conditions recorded during the experimental period
(Valea Calugareasca, 2017 year)

Sum of Sum of Monthly Number of

Month active useful average Rainfalls days with
temperature temperature temperature rainfall
January 0 0 -3.3 7.3 2
February 20.7 0.7 1.3 32.9 3
March 145.6 25.6 9.2 9.8 5
April 226.7 56.7 10.9 97.4 9
May 571.0 261.0 18.4 56.4 9
June 636.9 336.9 21.2 82.3 6
July 710.4 400.4 22.9 86.6 11
August 748.5 438.5 24.1 36.4 6
The growing
2893.5 1493.5 19.5 359.1 41
season
TOTAL 3059.8 1519.8 13.1 409.1 51

Table 2
Physical and hydrophobic soil indices within the experimental polygon

Depth Apparent The coeficient of The field capacity

cm wilting (CO) (CC)
density (DA)

% Cubic % Cubic
t/cubic meters meters/ meters/
hectare hectare
0-80 1.44 10.8 1244 19.9 2292

Depth Useful water Minimum limit Momentary reserve of

cm capacity (CU) 50% CU water in soil
% Cubic % Cubic % Cubic meters
meters/ meters/ hectare
hectare hectare
0-80 9.1 1048 15.4 1768 25.45 2932

The hydro physical soil (CO, CC) indices from the experimental polygon on
22.06.2017 highlight that the soil can store a volume of water (CU) of 1048 cubic
meters/hectare. Momentary water reserve in the soil is 2932 cubic meters/hectare,
resulting in a surplus of 1164 cubic meters/hectare. Summed amount of water

161
accumulated in the precipitation of a rainwater collector was 64.70 (32.35 l/vine),
resulting in an amount of 161.75 cubic meters/hectare. The irrigation norm used to
supplement the deficit was 231.25 cubic meters/hectare for variant 1-drip irrigation and
pluvial collector and 393 cubic meters/hectare for the variant 2- drip irrigation.
The analysis of the data obtained for the two irrigated and non-irrigated
variants shows a significant increase of soil microbiological activity in case of irrigated
variants at the sampling horizon of 0-30 and 30-60 centimeters (Table 3).

Table 3
The microbial load of soil in irrigated and non-irrigated variants
(Valea Calugareasca, 2017 year)

Variants Depth Total Other Actinomycete Yeasts + Nitrogen

cm number of Bacteria Fungus fixative
live bacteria
microorga-
nisms /
g soil % % % %
0-30 2673200 67.34 1.68 30.86 0.12
V1– irrigated
30-60 3448000 92.81 0.84 6.09 0.26
0-30 2906500 75.69 0.83 23.40 0.09
V2– irrigated
30-60 5979000 91.99 0.20 7.69 0.12
V3 – non 0-30 460000 84.78 4.35 6.52 4.35
irrigated 30-60 919200 84.86 1.22 10.01 3.92

The most numerous group of microorganisms in case of the two systems was
that of bacteria which represented 67.34-92.81% from the total number of viable
microorganisms.
In case of non-irrigated variant there was observed a reduction of the total
number of microorganisms with 17.21-26.66%, in case of the soil profile 0 - 30 cm and
with 15.37-15.83% in case of the soil profile of 30 - 60 cm.
Regarding Actinomycetes group, there was noticed a reduction with 55.56-
16.66% for the soil profile 0 - 30 cm and of 75-53.33% for the soil profile of 30 cm - 60
cm, compared to the irrigated variants.
The group of fungus was represented by 23.40%-30.86% for the total number
of microorganisms, in case of the irrigated variants, for the soil profile 0-30 cm, and in
area percent, less than 10% for the other variants.
Fungus belonging to the genus Penicillium, Aspergillus, Fusarium, Alternaria,
Cladosporium, Mucor, Verticillium, Phoma, Trichoderma and Gliocladium, were
identified.
The most representative genus for irrigated variants were Penicillium,
Cladosporium si Aspergillus.
The irrigation systems used in the study also have an important influence on
the physiological processes in the plant.
Regarding the photosynthesis process, the highest values were obtained for
the two irrigated variants (Figure 1).

162
 30
24,85 24,56
25
Photosynthesis
22,42
process intensity
20
(µmol CO2/m2/s)
15 Transpiration process
intensity (mmol
10 H2O/m2/s)
4,22 4,56 Respiration process
5 2,80 intensity (µmol
2,74 3,12 CO2/m2/s)
0 1,05

0 1 2 3 4
Figure1. The physiological processes in the plant

The influence of two irrigation systems compared to the non-irrigated control

on grape production and quality is shown in Table 4.

Table 4
The influence of experienced technological factors on
the production of grapes to Tamaioasa romaneasca variety

The Number The The Total acidity

average of production sugar
Variants weight of grapes
a grape g/l tartric
g Kg/ha g/l acid
V1-
drip irrigation and 204 6.9 1.408* 209,0 8.1
pluvial collector
V2- drip irrigation 202 6.8 1.374* 209,0 8.1
V3-non-irigated 168 7.0 1.177 201,0 8.5
DL 5% DL 1% DL 0.1%
0.18 0.25 0.31

Based on the data obtained from the mechanical analysis of the grapes, the
main urological indices characterizing the technological abilities of the grapes were
calculated (Table 5).
The higher values of the mechanical composition indices of the grapes both in
terms of the mechanical composition and the quality of the berry, allowing to obtaining
wines with good quality were obtained in case of irrigated variants.
Regarding the influence of the two irrigation systems on the grape yield, the
variance analysis emphasized a positive effect as compared with the non-irrigated
control.

163
 Table 5
Variation of the values of mechanical berry composition indices
depending on the irrigation system

Grape
Berry Composition Yield
Variants harvesting
index index berry index
index
V1-
drip irrigation and 77.9 57.8 7.5 7.5
pluvial collector
V2- drip irrigation 74.2 55.5 7.4 7.4
V3-non-irigated 73.4 52.6 7.4 6.7

The must sugar concentration depended to a lesser extent on the influence of

the irrigation systems.

CONCLUSIONS
The using of different system of irrigation are positive effects on plants, in
terms of the stimulation of the physiological processes in the plant, the increasing of
soil microbiological activity and the ensuring an optimal growth and yielding ratio.
The obtained results put into evidence the positive influence of irrigation, both
upon the grape yield and its quality, no matters what irrigation method was used. The
increasing yields of the grapes were obtained in comparison with the not irrigated
witness control, averagely ranging in between the limits of 19 - 20%, on the
background of a better accumulation of sugars in grapes.

REFERENCES
Pircalabu L., Serdinescu, 2004, Cercetãri privind utilizarea irigãrii subterane şi a
irigãrii prin picurare (prin conducte t-tape) în vederea combaterii stresului hidric la viţa
de vie. Anale ICDVV, vol. 14, .
Ojeda H., Saurin N., 2014, L‟irrigation de précision de la vigne: méthodes, outils et
stratégies pour maximiser la qualité et les rendements de la vendange en économisant
de l‟eau. Innovations Agronomiques 38 (2014), 97-108

164
 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
 Food produce process ing
UNIVERSITY OF CRAIO VA
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

THE BEHAVIOUR OF SOME PEACH VARIETIES (PRUNUS PERSICA L.)

BELONGING TO THE WORLD GERMOPLASM COLLECTION,
CONCERNING THEIR GROWTH AND FRUCTIFICATION IN CONDITIONS
OF BANAT PLAIN AREA
1* 2 3
Iordănescu Olimpia Alina , Cosmulescu Sina Niculina , Becherescu Alexandra ,
4 5 6
Murgu Viorica Adriana , Sala Florin , Scedei Daniela
1,3,6
Banat's University of Agricultural Science and Veterinary Medicine "King Michael I" from Timișoara,
Faculty and Horticulture and Forestry, Timișoara, Romania,
2
University of Craiova, Faculty of Horticulture, Craiova, Romania
4
Community Police, Timișoara, Romania
5
Banat's University of Agricultural Science and Veterinary Medicine "King Michael I" from Timișoara, Faculty
of Agriculture, Soil Science and Plant Nutrition, Timișoara, Romania
*Correspondence author. E-mail : [email protected]

Key words: biometric parameters, coefficient of variation, fruit quality indices, peach

ABSTRACT
To obtain higher fruit productions and qualitative fruits suppose in addition to applying
of superior technology culture a good zoning, more exactly knowledge of different behavior of
varieties in agreement with climatic and soil conditions in specific area. The biological material
was constituted by nine peach tree varieties from around the world. So, there were determined
varieties with a high Coefficient of variation (CV) for tree diameter CV = 48.10 ('Piros
Magdalena') to CV = 20.03 ('Poli'), tree height CV = 33.79 ('Gold Dust') to 6.86 ('Elbertina'),
number of short formation CV = 99.60 ('Poli') to CV = 16.86 ('Springold'), number of medium
formation CV = 74.32 ('Poli') to CV = 21.38 ('Springold'), number of long formation CV = 112.54
('Poli') to CV = 40.02 ('Yinquing'). Concerning quality fruit parameters, submitted high variance
for fruit weight CV = 47.89 to CV = 6.07 ('Sui Hui Hun' – GDRT), large diameter of fruits CV =
15.92 ('Sun Hui Hun') to CV = 3.45 (, 'GDRT'), high of fruits CV = 10.39 (Piros Magdalena) to CV
= 3.14 (Springold), index size CV = 14.44 to CV = 3.83 ('Sun Hui Hun' - 'GDRT').

INTRODUCTION
The peach tree (Prunus persica var. persica) originary from China is a highly
important species, being cultivated all over the world in different eco-climates and
using a large range of varieties, clones or local cultivars (Faust & Timon 1995; Zheng
et al. 2014; Wang et al. 2006). Due to its high importance, a series of studies were
developed on this species, starting from germplasm and its genetical diversity (Wang
et al. 2006; Verde et al. 2013; Meng & Peng 2015), to its physiological indices and
biometrical parameters (Quilot et al. 2002; Bregoli et al., 2006), its relation towards
ecological factors and stress (Zhang & Tian 2010; Pinochet et al. 2012), fruits‟ quality
indices (Bonora et al. 2013; Gasic et al. 2015; Mitre 2008), culture technology
elements and products‟ and bio-product‟s marketing (Bielamberg et al. 2009). The
physiological growth of peach trees estimated through the annual growth shoots,
through stock‟s biometrical parameters, trees‟ height, crown‟s diameter, etc. have been
165
studied in order to determine the general physiological status and the productive
potential of trees related to their age, climate and soil culture conditions and the culture
technology (Mounzer et al. 2008; Ikinci 2014; Basile et al. 2015). In relation to the
dynamics and specificity of nutrients for horticultural species and especially for
peaches, studies have been carried out to characterize some physiological indices,
biometric parameters and production in quantitative and qualitative terms
(Chatzitheodorou et al. 2004; Han et al. 2015; Iordănescu et al. 2014; Gruia et al.
2009). The balance between annual tree growth and fructification is controlled by
current crown formation (Dumitru et al. 2006) and fruiting pruning, by fertilization,
irrigation (Layne et al. 2002) and others, study models of trees‟ architecture and
physiology being made and used successfully in this respect (Xia et al. 2004; Allen et
al. 2005; Lopez et al. 2010; Bevacqua et al. 2012). Fruit size and quality indices of
peaches were studied in relation to the genetic potential of different genotypes (Monet
et al. 1996), with environmental and technology factors, nutrition and stress factors,
too. It is known that there are a number of interdependence relationships between
plant nutrition status, physiological and biometric parameters, fruit quality, for peach
tree and for other fruit species. (Bregoli et al. 2006; Jivan & Sala 2014). The quality of
fruits, products and by-products in horticultural species and peach has been studied in
relation to competitive relations with orchard grass (Tworkoski & Glenn, 2001), the
ripening moment (Ziosi et al. 2008), storage conditions (Wang et al. 2006). Considering
the importance of knowledge and management of fruit quality and production, different
models have been developed for the evaluation and estimation of some physiological
indices (Sala et al. 2015) and of some quantitative and qualitative aspects of peach
production (Lescouuret & Génard 2005; Zhang et la. 2009; Grechi et al. 2008; Lopez et
al. 2011).
The present study aimed at comparative analysis and characterization of some
peach varieties in terms of physiological indices, biometric parameters and specific
quality indicators of fruits.

MATERIAL AND METHOD

Location and experimental conditions. The research field is placed in the
fruit tree orchard belonging to the Didactic Station of Banat's University of Agricultural
Science and Veterinary Medicine from Timişoara, more exactly on the 2 ha parcel
cultivated with some peach and nectarine varieties that belong to the World Peach
Germplasm Collection. The research developed during the climatic conditions of 2012-
2014 years, the varieties being cultivated on a chernozem soil with average fertility.
Biologic material. The biological material consists of nine peach tree varieties
from around the world, as it follows: 'Poli', 'Sun Hui Hun', 'Yinquing', 'Piros Magdalena',
'Gold Dust', 'Eureka', 'July Elberta', 'Giala di Roma Tardiva', 'Elbertina' and 'Springold'
variety considered the experimental witness. The worldwide biological material was
introduced in our country and multiplied at SCDP Băneasa, from where it was bought
and was planted afterwards in the soil and climate conditions of Timișoara, in 2007.
The used planting distance was of 4 x 2 m, the peach varieties were grafted on wax
cherry, while the crown form system is free palmet. The culture technology is the
standard one used in peach tree orchards.

166
 Table 1

Climatic conditions during the observations

January February March April May June July August September October

Rainfall (mm)

2012 59.1 47.3 13.5 73.5 128.5 55.1 40.5 31 30.7 46.3

2013 43.7 39.8 31.4 83.2 115.8 113.4 40.2 50 90.2 50.4

2014 47.3 12.2 39.1 83.2 115.8 88.8 123.5 62.8 54 67.4

Temperature (ºC)

2012 1 -2.0 7.0 13.0 16.0 22.0 22.0 23.0 20.0 13.0

2013 1.0 4.0 5.0 13.0 17.0 20.0 22.0 23.0 15.0 14.0

2014 3.0 6.0 9.0 12.0 16.0 20.0 22.0 22.0 18.0 13.0

Studied parameters. Peach varieties have been studied in terms of

physiological indices and biometric parameters, expressing the physiological aspects
of trees‟ growth and development, thus: trunk‟s diameter (DT), tree‟s height (IP),
crown‟s diameter (DC), the number of short shoots (NFS), the length of short shoots
(LFS), the number of medium shoots (NFMD), the length of medium shoots (LFMD),
the number of long shoots (NFLD), the length of long shoots (LFLD). Fruits‟ quality was
evaluated through some specific indices: fruits‟ weight (GF), fruits‟ big diameter (DMF),
fruits‟ small diameter (Dm F), fruits‟ height (IF), fruits‟ size index (IMF), stone
percentage (PSF), sugar content (CZF), dry substance content (CSU).
Statistical analysis. Results obtained were analyzed by correlation and
regression in order to evaluate the interdependence between physiological indices,
biometrical parameters and fruits‟ quality indices. Relationships of interdependence
were described by second degree polynomial equations, the safety parameters used
2
being represented by the correlation coefficient R , parameter p and sample F.

RESULTS AND DISCUSSIONS

Results. Recorded data on vegetative indices, biometric parameters of trees,
and fruit quality indices for peach varieties revealed a correlation of these in relation to
the climatic conditions during the study period. At the same time, there were observed
differences between the peach varieties for each of the evaluated parameter, all the
collected data being presented in table 2. For trunk diameter parameter, Sui Hun Hun
variety had the lowest values (3.78±0.86 cm), being a variety with low vigour, while the
highest values were registered for Elbertina variety (5.62±0.84 cm). Trees‟ height gave
values that ranged from 164.11±8.63 cm for Sui Hun Hun variety and 274±8.87 cm
Elbertina variety. As for crown‟s diameter, the minimum values were obtained from
Piros Magdalena variety - 127.11±6.70 cm, whith the maximum values were measured
to Poli variety - 261.67±6.73 cm. The number of small shoots was lower for Elbertina
variety - 6.44±2.47 and higher for Eureka variety - 21.00±9.38. the length of small
shoots had values of 1.39±0.24 m for Springold variety and 2.30±0.26 m for Yinquing
variety. The number of medium shoots was lower for Piros Magdalena variety -
167
9.22±0.59 and higher for July Elberta variety 36.88±5.05. The length of medium shoots
was smaller for Springold variety - 10.61±0.30 cm and bigger for Elbertina variety -
18.22±4.59 cm. the number of long shoots ranged from 8.00±1.53 for Yinquing variety
and 27.66±3.38 for Elbertina variety, while their length had values of 26.78±1.31 cm
for Yinquing variety and 35.44±5.79 cm for Poli variety. The analysis of vegetative
indices and parameters values in relation to the climatic conditions revealed relations
of interdependence with rainfall. For the following parameters: trunk‟s diameter, trees‟
height, crown‟s diameter, for all peach varieties there were determined negative
correlations towards rainfall in May, for different signification levels, from very high (r =
-0.993 for July Elberta variety, trunk‟s diameter), to high level (r = -0.771 for Piros
Magdalena, trunk‟s diameter) and to average level (r = -0.686 for Eureka variety, trees‟
height). For the same parameter, during the other months of active vegetation (March–
October), with the August rainfall, the most constant correlations for all varieties and
the highest intensity were obtained (r =0.941–0.999). Within the other months of the
study period, differentiated manifestations of this interdependence were recorded.
Within the other studied vegetative parameters were recorded relations of
interdependence with the variable intensity rainfall, from very high (r =0.995, Poli
variety, the number of small shoots) to high significance (r =0.866, Elbertina variety,
length of small shoots), average significance (r =0.613, Eureka variety, length of small
shoots), and to low significance (r =0.354, July Elberta variety, number of long shoots).
Regarding the distribution of rainfall and the correlation level for the short, medium and
long shoots differentiation parameters and their dimensions, most of the varieties
correlated positively with the rainfall level in July, August and September, which shows
the sensitivity of the studied varieties to the lack of water in this period and the need for
irrigation to ensure normal vegetation conditions. The values of fruit quality indices
showed differentiated relationships of interdependence with average monthly rainfall
and temperatures over the study period. Dimensional parameters, such as weight,
height, big and small fruit diameter, correlated positively with precipitation in July and
August (r=0.997, Piros Magdalena variety, fruits‟ weight; r =0.999, Eureka variety,
fruits‟ big diameter; r = 0.999 Eureka and Piros Magdalena varieties, fruits‟ small
diameter; r =0.996, Piros Magdalena variety, fruits‟ height). Analyzing the correlation
coefficient values for all varieties and fruit dimensional quality indices, July to August
rainfall had a greater influence on the fruits. The fruit size index, as a synthetic index,
showed higher interdependence relations with the August rainfall (r =0.853 in July, r =
0.909 in August). Fruits‟ sugar content correlated negatively to the June-August
rainfall, with higher values for Sui Hun Hun variety in June (r = -0.891), Eureka variety
in July (r = -0.998) and Piros Magdalena and Gold Dust varieties in August (r = -0.893).
In contrast, for the period July-August, positive correlations were found for the dry fruit
substance and the monthly average temperature, with very significant correlation
values for Gold Dust variety (r = 0.891, July) and Giala di Roma variety (r = 0.987,
August).

168
 Table 2
Physiological indices, biometric parameters and fruit quality indices for the studied
peach varieties 2012 - 2014
Sui Piros Giala di
Yinquin Gold July Elbertin Springo
Poli Hun Magdale Eureka Roma
g Dust Elberta a ld
Hun na Tardiva

2012 3.97 2.30 2.40 3.03 3.83 4.10 2.40 3.07 4.00 3.40
DT
2013 5.23 3.77 3.13 4.13 5.47 4.67 4.50 4.33 6.03 5.50
2014 6.13 5.27 4.33 6.13 5.60 6.03 4.80 4.70 6.83 6.27
2012 208.33 150.33 216.67 151.00 180.67 192.00 190.33 188.67 260.33 182.67
IP
2013 215.00 162.00 226.00 163.33 187.00 198.33 217.00 201.67 271.33 201.00
2014 230.00 180.00 248.33 183.33 205.00 218.33 231.00 221.67 290.67 220.00
2012 250.00 175.50 200.33 116.00 181.67 143.33 188.17 160.83 192.83 149.00
DC
2013 261.67 188.17 212.67 126.17 195.50 152.20 207.67 173.50 208.50 160.33
2014 273.33 200.83 230.67 139.17 223.83 170.83 225.00 184.33 234.17 173.33
2012 4.00 3.00 12.67 11.67 18.00 15.33 6.67 10.00 4.67 8.00
NFS
2013 5.33 19.33 12.00 7.67 29.33 39.33 12.33 10.00 11.33 10.00
2014 17.00 16.00 11.33 11.00 7.33 8.33 17.33 2.00 3.33 9.33
2012 2.17 1.67 2.77 3.00 1.17 1.67 2.00 1.83 3.17 1.00
LFS
2013 1.83 1.67 2.23 1.67 1.47 1.50 2.00 2.00 2.17 1.83
2014 1.33 1.50 1.89 1.83 2.17 2.50 1.00 1.33 1.17 1.33
NFM 2012 11.67 17.67 16.00 9.00 15.00 11.33 29.67 13.00 16.00 11.33
D 2013 24.00 7.67 10.33 10.33 19.33 16.00 34.33 10.00 23.33 14.00
2014 36.67 11.00 15.00 8.33 23.33 25.67 46.67 15.67 18.00 14.00
2012 13.33 17.00 10.67 12.00 12.67 11.33 14.67 13.00 12.67 11.33
LFMD
2013 18.00 15.00 17.00 13.33 20.00 17.00 15.00 14.00 27.33 10.00
2014 16.67 18.00 16.00 14.67 15.33 18.00 13.67 15.00 14.67 10.50
2012 5.00 8.00 7.00 11.33 18.00 7.33 12.00 15.33 21.00 13.00
NFLD
2013 4.00 8.00 6.00 9.33 14.67 14.33 20.33 11.00 30.00 11.00
2014 26.33 13.67 11.00 14.67 26.33 21.67 42.00 29.00 32.00 15.00
2012 36.33 35.33 26.00 25.00 31.67 30.00 36.33 30.00 36.00 36.00
LFLD
2013 25.00 29.50 25.00 35.50 27.00 32.33 33.67 27.17 37.17 32.00
2014 45.00 36.00 29.33 29.00 36.00 34.33 33.67 38.00 30.00 35.00
2012 41.33 45.33 93.93 51.67 39.32 51.33 41.00 62.67 54.15 37.34
GF
2013 59.02 46.20 99.21 54.93 39.45 55.35 46.70 67.22 66.84 42.45
2014 58.86 100.46 82.93 91.84 65.13 86.03 55.54 63.14 73.70 59.90
2012 42.33 42.33 51.33 45.00 40.00 46.67 43.67 49.17 46.00 42.33
DMF
2013 48.67 43.67 52.39 46.00 41.50 47.07 44.44 50.00 61.67 43.39
2014 49.00 56.33 53.89 56.00 48.29 56.00 47.67 51.17 60.81 49.97
2012 40.67 40.67 50.33 42.67 38.33 45.67 42.00 45.00 44.33 40.33
Dm F
2013 46.67 45.83 50.72 43.33 40.17 46.00 43.16 47.33 58.00 42.27
2014 48.33 54.67 52.67 55.17 46.19 52.67 46.33 49.50 58.05 47.00
2012 43.67 47.00 57.33 45.33 40.67 45.67 42.67 47.00 36.00 44.67
IF
2013 48.00 49.17 59.11 46.33 42.67 43.33 43.39 47.50 36.00 45.44
2014 46.33 61.00 57.45 55.33 45.75 55.67 47.67 47.50 36.50 46.97
2012 42.22 43.33 53.00 44.33 39.66 46.00 42.77 47.05 42.11 42.44
IMF
2013 47.77 46.22 54.07 45.22 41.44 45.46 43.63 48.28 51.89 43.70
2014 47.89 57.33 54.66 55.50 46.74 54.78 47.22 49.38 51.78 47.97
PSF 2012 12.00 16.47 8.46 8.00 7.79 7.49 8.37 6.29 9.00 10.97
2013 11.50 16.11 8.23 9.80 7.79 7.40 8.22 6.45 8.80 11.03
169
 Sui Piros Giala di
Yinquin Gold July Elbertin Springo
Poli Hun Magdale Eureka Roma
g Dust Elberta a ld
Hun na Tardiva

2014 10.50 15.80 8.27 8.50 7.60 7.20 8.00 6.20 8.50 10.27
2012 9.16 9.61 13.89 13.08 13.93 14.31 10.17 14.07 10.67 10.03
CZF
2013 10.25 9.11 13.95 12.61 13.43 14.04 9.45 13.65 12.16 8.96
2014 11.55 9.53 10.60 10.46 11.13 10.56 10.70 11.63 8.41 10.13
2012 11.00 11.40 15.43 14.67 15.47 16.17 11.93 15.60 12.40 11.80
CSU
2013 12.00 10.93 15.50 14.23 15.00 15.57 11.26 15.20 13.80 10.79
2014 13.23 11.33 12.33 12.20 12.83 12.30 12.43 13.30 10.27 11.90

Discussions. This study evaluated the behavior of nine peach varieties in the
pedoclimatic conditions of the Banat Plain, in terms of indices and parameters
reflecting annual increases, biometric parameters of tree trunk and crown and fruit
quality indices. Similar studies of vegetative growth were also carried out in other
researches of different peach varieties and cultivars in relation to rootstocks and
pedoclimatic cultivation conditions (Weibel 2003; Forcada et al. 2014; Baciu et al.
2013). Gleen et al. (2011) studied different peach production systems in terms of yields
and economic values, interactions between tree density and architecture, cutting
strategies and soil management. DeJong et al. (1996) have developed a computer
simulation model on annual vegetative and generative peach growths. Based on the
environmental parameters, the model simulated the photosynthetic rate and the growth
of the vegetative and generative organs. The sensitivity of peach to water stress was
studied by Rahmati et al. (2015) in semiarid culture conditions, assessing annual
growth, fruit growth and rhythm of growth, and physiological indices directly involved,
such as photosynthesis, gas exchange, sweating. Annual vegetative increases are
very sensitive to moisture deficiency and reduce much more to water stress compared
to fruit growth (Egea et al. 2013). Peach roots were considered less affected under
water stress compared to annual shoots and fruits‟ growth (Hsiao and Xu 2000).
Lopez and DeJong (2007) found a significant influence of spring thermal
conditions during the blooming period and for 30 days after flowering on the annual
growth rate, the thermal values of the period being useful in estimating the fruit
production. Similar results on the relationship between climatic conditions and annual
increases and peach productivity were also reported by Atkinson et al. (2013). Studies
on the variation of peach growth and productivity were also performed by Matei et al.
(2012); Iordănescu et al. (2015) in relation to different varieties, the planting system
and organic and mineral fertilization, with positive correlations that are statistically safe.
Naor (2006) evaluated the relationship of peach tree in different stages of vegetation,
with the water stress caused by controlled irrigation. Regarding peach fruit quality
parameters and indices, many studies have observed these aspects in relation to
different varieties, environmental and technological factors (Fuentes-Pérez et al. 2014;
Cirilli et al. 2016) confirming the results obtained in this study.

CONCLUSIONS
As a result of the research conducted on lesser known and cultivated peach
varieties in our country under the pedoclimatic conditions of Banat Plain area, the
varieties can be grouped as follows: depending on the culture system, for intensive
170
orchards with high density of trees on the surface unit (Giala di Roma Tardiva, Eureka,
Gold Dust, Sun Hui Hun, Piros Magdalena), for intensive orchards with medium density
of trees on the surface unit (Yinquing și Poli) and for family orchards or gardens (July
Elberta and Elbertina); depending on the dominant fruiting shoots, Elbertina, Giala
di Roma Tardiva, July Elberta, Poli need extensive pruning, while Eureka and Sun Hui
Hun need less pruning; depending on fruit weight and size index value, Yinquing,
Sun Hui Hun, Piros Magdalena, Eureka have large fruits, for fresh consume, but also
for industrialization and Gold Dust, July Elberta, Poli have smaller fruits; depending on
sugar and dry substance content, Giala di Roma Tardiva, Eureka, Gold Dust,
Yinquing have a high content of these elements, while Sun Hui Hun, July Elberta,
Elbertina have a lower content.

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 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
 Food produce process ing
UNIVERSITY OF CRAIO VA
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

THE INFLUENCE OF THE PLANTING DISTANCE ON THE DEVELOPMENT

OF THE CHERRY ROOT SYSTEM

Ivanov I., Balan V., Balan P.

Agrarian State University of Moldavia

Correspondence author. E-mail: [email protected]:[email protected]

Key words: Cherry, variety, root, planting distance

ABSTRACT
This study presents the distribution of roots in the soil in commercial cherry orchards
(Prunus avium L.) depending on the planting distance. The study was carried out in the southern fruit
tree area of the Republic of Moldova on the Ferrovia cherry variety, aged 3 years, grafted on Gisela 6,
planted at the distance of 5x1.5 m, 5x2 m and 5x2.5 m and crowns after the Crown naturally
ameliorated with low volume. The weight and length of the roots were determined on horizontal
sections at 0-25 cm, 26-50 cm, 51-75 cm, 76-100 cm, 101-125 cm and 126-150 cm from the trunk, at
0- 20 cm, 21-40 cm, 41-60 cm and 61-80 cm. Each soil layer has been rooted, washed, dried, and
distributed in size categories: below 1 mm, 1-3 mm, 3-5 mm, over 5 mm. The length of roots is
determined by fibrous roots (95.4-95.8%), and mass - by skeletal roots (65.6-69.3%). Most roots after
length (69.8-82.9%) and after meal (88.8-92.8%) are distributed on a depth of 0-60 cm.

INTRODUCTION
The density of cherry trees in modern orchards is constantly changing due to
vegetal rootstocks, new varieties and new tree management systems. (Whiting & amp;
Ophardt 2005, Calabro et al., 2009, Balan 2015). Considering that the planting distance of
cherry trees is determined by the vigor of the variety and of the rootstock, the crown shape,
the soil fertility and the equipment, it has been felt that it is necessary to study the
development of the root system and the distribution of the roots in the soil at different
planting distances of cherry trees. (Long et al., 2014; Musacchi & Serra, 2015).

MATERIAL AND METHODS

The experience was organized in the southern fruit tree area of the Republic of
Moldova in the spring of 2009 at S.R.L. Terra Vitis in the village of Burlacu Cahul district with
cherry rootstock Gisela 6 grafted with cherry varieties Bigarreau Burlat, Ferrovia, Lapins.
Planting distances were 5x1.5 m, 5x2 m, 5x2.5 m. The low crown shape was used: Crown
naturally ameliorated with low volume, and Slender Spindle.
The root system development study was carried out on the Ferrovia variety formed
after the reduced-volume natural crown. Three trees were chosen for the study, in each
variant, with uniform increases, after the diameter of the trunk, the height and diameter of the
crown.
The root mass and form were studied at the end of vegetation (1/4 of the tree nutrition
surface was dug) in terms of their spreading in the soil layers vertically and horizontally. The
weight and length of the roots were determined on horizontal sections at 0-25 cm, 26-50 cm,
51-75 cm, 76-100 cm, 101-125 cm and 126-150 cm from the trunk, at 0- 20 cm, 21-40 cm,
175
41-60 cm and 61-80 cm. In each layer of soil, the roots were selected, washed, dried, and
distributed in size categories: below 1 mm, 1-3 mm, 3-5 mm, over 5 mm.

RESULTS AND DISCUSSIONS

In order to obtain large and regular productions in intensive plantations, it is
necessary to guide the growth of both crown and roots (Babuc 2012). Analyzing the data
obtained (Table 1), it is found that cherry trees of the Ferrovia variety at the planting distance of
5x1,5 m have the root system less developed than at the larger distances taken in the study.

Table 1
The influence of the planting distance of the root system development Ferrovia cherry
variety grafted on Gisela 6, age of 3 years
Planting Length of roots, cm/tree Mass of roots, g/tree
distance, Under Over Under Over
1-3 mm 3-5 mm 1-3 mm 3-5 mm
m 1mm 5mm 1mm 5mm
5x1,5 17240 29320 1198 1032 76,8 130,2 105,6 290,0
5x2 25436 29633 1508 1084 83,2 157,5 142,4 336,4
5x2,5 21712 35665 1240 1248 73,6 154,6 156,0 359,2
Media 21462 31539 1315 1121 77,8 147,4 134,6 328,5
By increasing the density of the trees to a surface unit from 800 to 1333 tree/ha, the
root length decreases from 59867 cm to 48790 cm. The reduction of the root system in the
over grown plantations takes place on roots with a diameter of less than 1 mm and a 1- 3
mm. 3-year-old cherry trees make up 38.7% of roots below 1 mm in diameter, 56.9% in
diameter 1-3 mm and only 4.4% in diameters over 3 mm in diameter.
The weight of the roots also varies according to the planting distance. The largest
root mass was recorded at the planting distance of 5x2.5 m and consisted of 743 g/tree. The
size of the root mass is based on roots with a diameter of more than 3 mm. From the values
shown, 67.3% make up the roots with a diameter of over 3 mm, 21.4% of 1-3 mm and 11.3%
of the diameter below 1 mm of the total mass. So, the root length of the root system
increases from the diameter of the fibrous roots by less than 3 mm in diameter, and the mass
- from the skeletal and semi-skeletal roots with a diameter of more than 3 mm.
Analyzing the distribution of the roots in depth (Table 2) it can be emphasized that
most of the roots, both in length and after mass, are found in the soil layer 0-60 cm. Thus, 3
years old cherry trees at the planting distance of 5x1.5 m in this layer are placed 82.9% by
length and 72.7% by mass. Analogously, the roots and the other planting distances studied
are distributed in depth. The highest density of fibrous, skeletal and semi-skeleton roots was
recorded in the 21-60 cm soil layer.
These roots are the basis of feeding the trees and have a more directionally less
horizontal (Babuc 2012). At the depth of 0-20 cm and 61-80 cm, a small amount of roots is
placed. Thus, at the planting distance of 5x1,5 m, the total length of the deep roots is placed
as follows: 0-20 cm - 5,3%; 21-40 cm - 32.0%; 41-60 cm - 45.6%; 61-80 cm - 17.1%. The
mass of the roots in depth changes in analogy to the length.
It has been found that in young cherry trees in the 0-20 cm layer there is an
insignificant amount of roots up to 3 mm in diameter and missing roots with a diameter of
more than 3 mm. These findings lead to the conclusion that the soil works in young orchards
are made superficially up to the depth of 15-20 cm, due to both the cutting of the roots
caused by the repeated works and the frequent summer drying of the superficial soil layer.
The data presented demonstrate that the planting distances taken in the study
practically do not influence the distribution of the roots in depth to the cherry trees of 3 years
old.
Most of the skeleton roots are found in superficial soil layers, and as the depth

176
increases, it shrinks abruptly. The deep distribution of the fibrous roots is more uniform than
the skeletal roots.
Table 2
Influence of the planting distance on the distribution of roots in the soil in the Ferrovia variety,
grafted on Gisela 6, 3 years old.

Soil Length of roots, cm/tree Mass of roots, g/tree

layer, Under Over Under Over
1-3 mm 3-5 mm 1-3 mm 3-5 mm
cm 1mm 5mm 1mm 5mm
Planting distance, 5x1,5 m
0-20 1192 1413 - - 4,0 2,6 - -
21-40 7434 7393 540 248 25,2 28,4 54,8 132,4
41-60 6490 14600 374 784 22,0 81,2 50,8 157,6
61-80 2124 5914 284 - 25,6 18,0 - -
Total 17240 29320 1198 1032 76,8 130,2 105,6 290,0
Planting distance, 5x2 m
0-20 3074 2098 - - 7,6 9,1 - -
21-40 7788 8409 844 312 26,4 36,4 70,8 68,4
41-60 10916 11550 476 772 36,8 86,4 59,2 268,0
61-80 3658 7576 188 - 12,4 25,6 12,4 -
Total 25436 29633 1508 1084 83,2 157,5 142,4 336,4
Planting distance, 5x2,5 m
0-20 1298 645 - - 4,4 2,8 - -
21-40 7080 7299 440 288 24,0 31,6 53,6 132,0
41-60 7316 15708 748 960 24,8 68,0 91,6 227,2
61-80 4484 10720 52 - 15,2 46,4 10,8 -
81-100 1534 1293 - - 5,2 5,8 - -
Total 21712 35665 1240 1248 73,6 154,6 156 359,2
The 3-year cherry tree root system did not occupy the entire nutritional area for
them (Table 3). Most roots, both in length and after the meal, are at a distance of 0-75 cm
from the trunk.
In this area, at the planting distance of 5x1,5 m, 65,2% of the length and 82,6% of
the root mass are placed at the distance of 5x2 m respectively, 66,2% and 81,9%, and at
the distance of 5x2,5 m respectively 60.9% and 71.3%. If we analyze the data presented in
the table and figure it is found that all the planting distances the length and the root mass
decrease from the trunk to the periphery.
The root mass decreases more suddenly than their length. Decreasing the
distance between trees in a row helps to increase the amount of roots around the trunk (0-
50 cm), reaching 47.2% of the length and 55.5% of the mass at a distance of 5x1.5 m.
Table 3
Influence of the planting distance on the distribution of the roots in the soil in the Ferrovia
variety, grafted on Gisela 6, 3 years old.
Length of roots, cm/tree Mass of roots, g/tree
Planting
Under 3-5 Over 5 Under Over 5
distance, m 1-3 mm 1-3 mm 3-5 mm
1mm mm mm 1mm mm
Planting distance, 5x1,5 m
0-25 4130 7300 160 364 10,4 30,2 5,2 100,5
26-50 4248 5545 712 496 14,4 24,0 92,4 52,4
51-75 2124 6423 108 80 7,2 28,8 24,2 100,4
76-100 2018 2495 48 92 6,8 10,8 4,0 32,4
101-125 2478 3861 - - 8,4 16,8 - -
126-150 2235 3696 - - 7,6 16,0 - -
Amount 17233 29320 1028 1032 54,8 126,6 125,8 285,7

177
 Planting distance, 5x2 m
0-25 4130 7393 128 312 14,0 29,0 9,2 80,4
26-50 6018 6376 712 584 20,4 27,6 89,6 212,0
51-75 2832 8010 88 112 9,6 44,4 19,2 16,0
76-100 4248 3327 180 76 14,4 14,4 24,4 28,0
101-125 4956 2772 - - 16,8 12,0 - -
126-150 2360 756 - - 8,0 7,6 - -
Amount 24544 28634 1108 1084 83,2 135 142,4 336,4
Planting distance, 5x2,5 m
0-25 5664 9980 120 468 19,2 43,2 10,0 192,0
26-50 4484 6375 620 332 15,2 17,6 82,4 56,0
51-75 2714 5544 160 108 9,2 24,0 14,0 40,0
76-100 2832 2679 60 120 9,6 11,6 8,0 12,4
101-125 2596 3972 220 180 8,8 16,8 24,4 48,0
126-150 3422 7207 60 40 11,6 31,2 17,2 10,8
Amount 21712 35757 1240 1248 73,6 144,4 156,0 359,2

CONCLUSIONS
The roots of the cherry trees, grafted on the Gisela 6 rootstock, and shaped like a
reduced-sized natural crown, advance from the trunk radially evenly in all directions. As the
distance between tree trees increases, the length and root mass increase and their density in
the soil decreases. The length of roots is determined by fibrous roots (95.4-95.8%), and the
mass - of the skeleton roots (65.6-69.3%). The majority of the roots by length (69.8-82.9%)
and after the meal (88.8-92.8%) are distributed over a depth of 0-60 cm.
The extension and distribution in the soil of the radicular system allow to make the
following findings regarding some agro technical works in modern cherry plantations: In
young plantations with the soil maintained as a field, the soil loosening works can be
executed up to the depth of 15- 20 cm with the risk of cutting at most 5.3% of the roots by
length and 1.1% by mass with a diameter of up to 3 mm; The young trees, the roots near the
trunk (up to 75 cm) are higher in the superficial layer of 0-20 cm, it is better to avoid the
proximity of tree rows to less than 75 cm.
The administration of nitrogen fertilizers on the row of trees, on a 1.5 m wide strip, in
that the fibrous roots are denser in this sector of the nutrition of the trees. The migration of
phosphorus and potassium into the soil, being difficult, the administration of these fertilizers is
incorporated into the soil at a distance of 50-75 cm at a depth of 20 cm. Determining the
depth (need) of water supply to the soil must be the depth at which most roots are distributed,
especially 0-60 cm.

REFERENCES
Babuc, V. Pomicultura. Chişinău: Tipografia Centrală 2012, 662 p. ISBN 978-9975-53-
067.
Balan, V. Formarea coroanei după sistema „Fus subţire” în plantaţiile intensive de cireş.
Pomicultura, Viticultura şi Vinificaţia, Republica Moldova. Nr. 1 (55). Chişinău, 2015, p 18-21
Calabro J. M, Spotts R. A. and Grove G. G. (2009). Effect of Training System,
Rootstock, and Cultivar on Sweet Cherry Powdery Mildew Foliar Infections.
HortSciense, vol, 44: 481-482
Long L,E., Long M., Peşteanu A., and Gudumac E. (2014). Producerea Cireşelor.
Manual tehnologic. Chişinău, 2014, 262p.
Musacchi S., Gagliardi F. and Serra S. (2015). New Training Systems for High-
density Planting of Sweet Cherry. HortScience, January, vol 50, no. 1: 59-67.
Whiting M. D., Lang G. and Ophardt D. (2005). Rootstock and Training System Affect
Sweet Cherry Growth, Yield, and Fruit Quality. HortScience, vol 40, no. 3: 582-586.
178
 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
 Food produce process ing
UNIVERSITY OF CRAIO VA
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

EFFECT OF EXTRACTION TEMPERATURES AND SOLVENT ON THE

ANTIOXIDANT POTENTIAL OF GREEN ARABICA AND ROBUSTA
COFFEE BEANS
1 2 2
Kamvoukou Christina- Anna , Gougoulias Nikolaos *, Giannoulis Kyriakos ,
2 2 2
Papachatzis Alexandros , Papathanasiou Theodoros , Malamis Asterios
1
School of Pharmacy, Aristotle University of Thessaloniki, Greece
2
Department of Agricultural Technology, Technological Educational Institute of Thessaly, 41110 Larissa,
Greece
*Correspondence author. Email: [email protected]

Key words: Antioxidant activity FRAP; Antiradical activity DPPH; Coffee; Total phenols

ABSTRACT
Two different varieties green coffee (Robusta and Arabica) were studied for total
polyphenol content and for antioxidant properties. Different solvents and temperature extraction
conditions were employed. The total phenol content ranged from 4.73 to 60.31 mg (GAE)/g DW,
the antiradical activity DPPH• ranged from 26.2 to 158.1 µmol (Trolox)/g DW and the antioxidant
activity FRAP ranged from 115.8 to 811.9 µmol (Trolox)/g DW. Extraction with 80% aqueous
ethanol at 45oC temperature proved to be a optimum method to extract phenolic antioxidant
compounds from green coffee. The Robusta variety exert higher antioxidant activity FRAP and
higher antiradical activity (DPPH•), compared to the Arabica variety, for all extraction treatments.
The extracts green coffee can find applications as food additives, either in the form of powder or
as extracts highly concentrated.

INTRODUCTION
The reactive oxygen and nitrogen containing free radicals (ROS and RNS),
formed in the human cells as strong oxidants, can cause oxidative damage in the
biological molecules. These changes of the biological molecules are the basis for many
diseases (Halliwell 1991). Epidemiological studies demonstrate that the oxidative
destruction of biomolecules can be reduced by using endogenous and exogenous
antioxidants such as polyphenols, etc.(Pellegrini et al. 2003). They‟re in a diet; reduce
the risk of cardiovascular and some cancer diseases (Manach et al. 2004).
The coffee plant, is an evergreen shrub, where it belongs to the family
Rubiaceae, while when cultivated reaches about 2.5 meters high. Coffee contains a
tremendous number of phenolic compounds; the most abundant phenolic compounds
are chlorogenic acids, ferulic acid, tannins, proanthocyanidins, lignans, and
tocopherols, which exert strong antioxidant effect (Borelli et al. 2002, Farah &
Donangelo 2006, Gonzalez 2001). Other compounds in coffee affect its color and
aroma (Cammerer & Kroh 2006, Lopez-Galilea et al. 2006).
Studies have shown that coffee consumption is related with the reduction of
liver cancer, parkinson's disease, alzheimer's disease and the prevention of diabetes
mellitus type 2 (Dοrea & Costa 2005, Hidgon & Frei 2006, Lopez-Garcia et al. 2006).

179
 The aim of the present study is to determine the content of phenolic
compounds and the antioxidant activity, of the green Robusta and Arabica coffee
beans at different solvents and temperature extraction conditions.

MATERIAL AND METHODS

Sample preparation:
Two, commercial available, species of green coffee were purchased from the
local market in Larisa, Robusta coffee beans and Arabica coffee beans. The green
coffee were dried in a dark place at room temperature, finely ground and kept at 4oC.
The sample was extracted using four different conditions.

Extraction procedures:
(i) 4 g of the finely ground sample were 2-fold treated by 25 ml 80% aqueous
ethanol at 45 oC. At first treatment the samples were incubated for 4h in the extractant
o
at stirring and the second one continued stirring for 2h at 45 C. The extract was
gathered after centrifugation and filtration through Whatman No. 4 paper.
(ii) 4 g of the finely ground sample were 2-fold treated by 25 ml 80% aqueous
ethanol at ambient temperature. At first treatment the samples were incubated for 24 h
in the extractant at stirring and the second one continued stirring for 2h at ambient
temperature. The extract was gathered after centrifugation and filtration through
Whatman No. 4 paper.
(iii) 4 g of the finely ground sample was extracted with 50mL of boiling water at
150 rpm for 30 min. The extract was gathered after centrifugation and filtration through
Whatman No. 4 paper.
(iv) 4 g of the finely ground sample was extracted with 50mL of water at room
temperature for 24 h and filtered through Whatman No. 4 paper.

Methods of analyses:
Determination of total polyphenolics (TP). Total polyphenolic content was
determined with the Folin-Ciocalteu (F.-C.) reagent according to the method by
(Singleton and Rossi 1965) using the microvariant proposed by (Badenschneider et al.
2015) and the results were expressed as gallic acid equivalent (GAE) in mg/g dry
weight.
Determination of ferric reducing antioxidant power (FRAP). The antioxidant
activity of the methanol extracts was determined on the basis of the method by (Benzie
and Strain 1999) and was expressed as Trolox equivalent (TEAC) in µmol /g dry
weight.
Determination of antiradical activity (DPPH•). The antiradical activity of the
methanol extracts was determined according to the method by (Brand-Williams et al.
1995) using the stable free radical 2,2′-diphenyl-1-picrylhydrazyl (DPPH•). The activity
was evaluated as Trolox equivalent (TEAC) in μmol/g dry weight.

Statistical analysis:
Data were analyzed using the MINITAB (Ryan et al. 2005) statistical package.
The experiment had four replications. Analysis of variance was used to assess
treatment effects. Mean separation was made using Tukey‟s test when significant
differences (P=0.05) between treatments were found.

180
 RESULTS AND DISCUSSIONS
The extraction of the green coffee with different solvents and different
temperatures results in the extraction of phenol compounds which vary between 4.73
and 60.31 mg (GAE)/g DW (Table 1). In all treatments studied, the green coffee
Robusta characterised by higher TP content compared to green coffee Arabica. Our
results are in agreement with the data obtained by other authors who have established
differences in total phenols content in the different green coffee species (Hudáková et
al. 2016, Odžaković et al. 2016, Pérez-Hernández et al. 2012).
The use of solvents at high temperatures, compared to the respective solvents
at room temperature, showed a higher content of total phenols in the green coffee
Robusta and Arabica (Table 1). Extraction yield of total phenols in the green coffee
Robusta and Arabica shows the following order:
80% aqueous ethanol (45oC) > 80% aqueous ethanol (room temperature >
boiling water > Water (room temperature).

Table 1
Total phenols content of green Arabica and Robusta coffee beans

Solvent Conditions TP – (mg GAE/g) DW

Robusta Arabica
80% Aqueous ethanol 45oC 60.31a 44.53b
80% Aqueous ethanol Room temperature 39.01c 26.27d
Water Boiling water 16.86e 7.62f
Water Room temperature 12.79g 4.73i

For each extraction treatments characteristics with the same letter of table do
not differ significantly according to the Tukey‟s test (P=0.05).

The antiradical activity (DPPH•) of the two varieties green coffee studied
ranges from 26.2 to 158.1 µmol (Trolox)/g DW (Table 2). The use of solvent 80%
aqueous ethanol at temperature conditions 45oC exert the higher antiradical activity
(DPPH•) with 158.1 μmol (Trolox)/g DW, in the green coffee Robusta. On the contrary,
the use of water as solvent at room temperature exert the lower antiradical activity
(DPPH•) with 26.2 μmol (Trolox)/g DW, in the green coffee Arabica.
The Robusta variety exerts the higher activity, compared to the Arabica
variety, for all extraction treatments. The variation in the antiradical activity (DPPH•)
reported and by other authors for the green Arabica and Robusta coffee beans
(Kiattisin et al. 2016, Sentkowska et al. 2016).
The antioxidative assay FRAP of the two varieties green coffee studied ranges
from 115.8 to 811.9 µmol (Trolox)/g DW (Table 3). Extraction solvent 80% aqueous
ethanol at temperature conditions 45oC in the green coffee Robusta, exert the higher
antioxidant activity FRAP with 811.9 μmol (Trolox)/g DW. On the contrary, extraction
solvent water at room temperature in the green coffee Arabica, exert the lower
antioxidant activity FRAP with 115.8 μmol (Trolox)/g DW (Table 3).

181
 Table 2
Antiradical activity DPPH• of green Arabica and Robusta coffee beans

Solvent Conditions DPPH• - (μmol Trolox /g) DW

Robusta Arabica
o
80% Aqueous ethanol 45 C 158.1a 104.8c
80% Aqueous ethanol Room temperature 120.1b 82.3d
Water Boiling water 93.6e 63.9f
Water Room temperature 39.0g 26.2i
For each extraction treatments characteristics with the same letter of table do
not differ significantly according to the Tukey‟s test (P=0.05).

The Robusta variety exert higher activity (as Trolox equivalent) compared to
the Arabica variety, for all extraction treatments (Table 3). The variation in the
antioxidant activity FRAP reported and by other authors for the green coffee varieties
(Pokorná et al. 2015).

Table 3
Antioxidant activity FRAP of green Arabica and Robusta coffee beans

Solvent Conditions FRAP- (μmol Trolox /g) DW

Robusta Arabica
80% Aqueous ethanol 45oC 811.9a 763.2b
80% Aqueous ethanol Room temperature 615.2c 510.0d
Water Boiling water 301.5e 239.7f
Water Room temperature 135.3g 115.8i

For each extraction treatments characteristics with the same letter of table do
not differ significantly according to the Tukey‟s test (P=0.05).

CONCLUSIONS
The results obtained showed differences in total phenols composition in the
green Robusta and Arabica coffee beans. Phenolic compounds, antioxidant and
antiradical activity of green coffee depends strongly on the variety.
The higher extraction temperatures at each solvent increased the extraction
yield of total phenols in the green coffee Robusta and Arabica.
Green Robusta and Arabica coffee beans are a source of bioactive
components that could be included in functional foods composition. The extracts green
coffee can find applications as food additives, either in the form of powder or as
extracts highly concentrated.

REFERENCES
Baderschneider B., Luthria D., Waterhouse A.L., Winterhalter P., 2015,
Antioxidants in white wine (cv. Riesling): I. Comparison of different testing methods for
antioxidant activity. VITIS-Journal of Grapevine Research, 38(3), 127-131.
Benzie I.F., Strain J.J., 1999, Ferric reducing/antioxidant power assay: direct
measure of total antioxidant activity of biological fluids and modified version for
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simultaneous measurement of total antioxidant power and ascorbic
acid concentration. Methods in enzymology, (299), 15-27.
Borrelli R.C., Visconti A., Mennella C., Anese M., Fogliano V., 2002, Chemical
characterization and antioxidant properties of coffee melanoidins. Journal of
Agricultural and Food Chemistry, 50(22), 6527-6533.
Brand-Williams W., Cuvelier M.E., Berset C.L.W.T., 1995, Use of a Free Radical
Method to Evaluate Antioxidant Activity. LWT- Food Science and Technology, 28(1),
25-30.
Cämmerer B., Kroh, L.W., 2006, Antioxidant activity of coffee brews. European
Food Research and Technology, 223(4), 469-474.
Dórea J.G., da Costa, T.H.M., 2005, Is coffee a functional food?. British Journal of
Nutrition, 93(06), 773-782.
Farah A., Donangelo C.M., 2006, Phenolic compounds in coffee. Brazilian Journal
of Plant Physiology, 18(1), 23-36.
González A.G., Pablos F., Martın M.J., León-Camacho M., Valdenebro M.S.,
2001, HPLC analysis of tocopherols and triglycerides in coffee and their use as
authentication parameters. Food Chemistry, 73(1), 93-101.
Halliwell B., 1991, Reactive oxygen species in living systems: source,
biochemistry, and role in human disease. The American journal of medicine, 91(3),
S14-S2.
Higdon J.V., Frei B., 2006, Coffee and health: a review of recent human research.
Critical reviews in food science and nutrition, 46(2), 101-123.
Hudáková J., Marcinčáková D., Legáth J., 2016, Study of Antioxidant Effects of
Selected Types of Coffee. Folia Veterinaria, 60(3), 34-38.
Kiattisin K., Nantarat T., Leelapornpisid P., 2016., Evaluation of antioxidant and
anti-tyrosinase activities as well as stability of green and roasted coffee bean extracts
from Coffea arabica and Coffea canephora grown in Thailand. Journal of
Pharmacognosy and Phytotherapy, 8(10), 182-192.
Lopez-Garcia E., van Dam R.M., Willett W.C., Rimm E.B., Manson J.E., Stampfer
M.J., Rexrode K.M., Hu F.B., 2006, Coffee consumption and coronary heart disease in
men and women. Circulation, 113(17), 2045-2053.
López-Galilea I., Fournier N., Cid C., Guichard E., 2006, Changes in headspace
volatile concentrations of coffee brews caused by the roasting process and the brewing
procedure. Journal of Agricultural and Food Chemistry, 54(22), 8560-8566.
Manach C., Scalbert A., Morand C., Rémésy C., Jiménez L., 2004, Polyphenols:
food sources and bioavailability. The American journal of clinical nutrition, 79(5), 727-
747.
Odžaković B., Džinić N., Kukrić Z., Grujić S., 2016, Effect of roasting degree
on the antioxidant activity of different Arabica coffee quality classes. Acta Scientiarum
Polonorum Technologia Alimentaria, 15(4), 409-417.
Pellegrini N., Serafini M., Colombi B., Del Rio D., Salvatore S., Bianchi M.,
Brighenti F., 2003, Total antioxidant capacity of plant foods, beverages and oils
consumed in Italy assessed by three different in vitro assays. The Journal of nutrition,
133(9), 2812-2819.
Pérez-Hernández L.M., Chávez-Quiroz K., Medina-Juárez L.Á., Gámez Meza
N., 2012, Phenolic characterization, melanoidins, and antioxidant activity of some
commercial coffees from Coffea arabica and Coffea canephora. Journal of the Mexican
Chemical Society, 56(4), 430-435.
Pokorná J., Venskutonis P.R., Kraujalyte V., Kraujalis P., Dvořák P., Tremlová B.,
Kopřiva V., Ošťádalová M., 2015, Comparison of different methods of antioxidant

183
activity evaluation of green and roast C. Arabica and C. Robusta coffee beans. Acta
Alimentaria, 44(3), 454-460.
Ryan B.F., Joiner B.L., Cryer J.D., 2005, MINITAB Handbook: Updated for release
14, 5th edition.
Sentkowska A., Jeszka-Skowron M., Pyrzynska K., 2016, Comparative Studies
on the Antioxidant Properties of Different Green Coffee Extracts. MOJ Food
Processing & Technology, 3(2), 00071.
Singleton V.L., Rossi J.A., 1965, Colorimetry of total phenolics with
phosphomolybdic - phosphotungstic acid reagents. American journal of Enology
and Viticulture, 16(3), 144-158.

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 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
 Food produce process ing
UNIVERSITY OF CRAIO VA
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

EFFECT OF CYCOGAN ON GROWTH AND FLOWERING OF

BOUGAINVILLEA SPP.

Mandă Manuela1*, Nicu Carmen1

1*
University of Craiova, Craiova, e-mail: [email protected]

Key words: Bougainvillea, Cycogan, control, growth, flowering

ABSTRACT
The effect of growth inhibitor treatments on the growth and development of
Bougainvillea plants has been studied by many researchers, and the results indicate that
different species and varieties react differently to their application. The main objective of this
work was to investigate the effects of Cycogan treatments on the growth and development of
three species of Bougainvillea sp. The best results in terms of the retardant effect of Cycogan on
the plants were obtained in: B. buttiana "Raspberry Ice" - Cycogan 0.1% and 0.2%; B.
spectabilis "Orange King" - Cycogan 0.1% and 0.2%; B. buttiana "Alba" - Cycogan 0.1%; B.
spectabilis - Cycogan - 0.2%; B. glabra - Cycogan 0.2% and 0.3%. The flowering was positive
influenced by Cycogan treatments in B. glabra, B. spectabilis "Orange King" and B.
buttiana "Alba".

INTODUCTION
The Bougainvillea genus, originally from South America, is from the
Nyctaginaceae family and includes 14 species, of which the following three are of
particular interest: Bougainvillea spectabilis Willdenow, B. glabra Choisy and B.
peruviana Humboldt and Bonpland. Bougainvillea can be a shrub (B. peruviana H. &
P.), a small tree (B. praecox Griseb.) or a liana (B. spectabilis Willd., B. glabra Choisy)
(Randhawa & Mukhopadhyay 1986, Kobayashi 2007).
Bougainvillea has green or variegated leaves; the flowers are small and
tubular, usually white or yellow, supported by three persistent, brightly coloured bracts,
these being the main decorative element (Probert et al 1993).
Bougainvillea is a very popular evergreen landscape plant, used for decorating
pergolas, fences, walls, buildings, but also as a plant in pots or suspended baskets.
Bougainvillea x buttiana and B. glabra are ideal for topiary art and bonsai because they
are more compact (Cirillo et al. 2014, Whistler 2000). In our country, Bougainvillea can
be used in the decoration of spacious, well-lit interiors, but it is also present in gardens,
balconies and terraces during the warm season. Because of the vigorous growth habit,
bougainvilea needs a lot of labor-intensive pruning when being used as a potted plant.
PGRs could be alternatives to frequent pruning on bougainvillea to reduce labor costs
(Jain et al. 2014). The effect of growth inhibitor treatments on the growth and
development of Bougainvillea plants has been studied by many researchers, and the
results indicate that different species and varieties react differently to their application
(Shao et al. 2006, Saifuddin et al. 2009b, Jain et al. 2016). Thus, the treatment with
ethephon stimulated the Bougainvillea flowering (Fang-Yin & Chang, 2011). In the
185
"San Diego Red" variety, the CCC and B-9 treatments proved to be effective in
stimulating flowering in a short daytime period (Hackett & Sachs 1967). Aldrich &
Norcini (1996) studied the effect of PGRs on the production of Bougainvillea "Barbara
Karst". Foliar treatments with uniconazole (10 ppm), maleic hydrazide (2808 ppm),
daminozide (5000 ppm) and paclobutrazol 200 ppm) were applied. The PGR treatment
has not actually slowed the growth or has not increased the quality or marketability of
Bougainvillea plants "Barbara Karst". Moreover, the daminozide reduced the number of
ramifications, and the maleic hydrazide was phytotoxic.
Based on these considerations, the present study aims to present the effect of
Cycogan (1000, 2000, 3000 ppm) treatments on the growth and development of three
species of Bougainvillea spp.

MATERIALS AND METHOD

The biological material was represented by rooted cuttings of 3 species of the
Bougainvillea genus, namely: B. glabra, B. spectabilis, B. spectabilis "Orange King"
and B. buttiana with the "Alba" and "Raspberry Ice" varieties existing in the Floriculture
discipline collection.
Cycogan 400 LC 100 ml is a growth regulator that acts on plant as a growth
inhibitor but it is capable of producing a wide range of physiological changes in
different plant species and varieties.
The experiment was placed in the didactic greenhouse of the Floriculture
discipline of the A.I. Buia Botanical Garden from Craiova on 22.05.2015. The
greenhouse is temperate, with average temperatures ranging between 18 ºC and
20ºC. Two experimental factors were considered: Factor A - variety with 5 graduations:
a1. B. glabra, a2. B. spectabilis, a3. B. spectabilis "Orange King",a4. B. buttiana
"Alba", a5. B. buttiana "Raspberry Ice"; Factor B – the treatments with different
concentrations of Cycogan (C), with 4 graduations: b1. Water (Ct untreated); b2. C
0.1% (1000 ppm); b3. C 0.2% (2000 ppm); b4. C 0.3% (3000 ppm). The combination of
the two factors resulted in 20 experimental variants.: V11(Ct1) - B. buttiana ”Raspberry
Ice”/w; V12 - B. buttiana ”Raspberry Ice”/C 0.1%; V13 - B. buttiana ”Raspberry Ice”/C
0.2%; V14 - B. buttiana ”Raspberry Ice”C 0.3%; V21 (Ct 2) - B. spectabilis ”Orange
king”/w; V22 - B. spectabilis ”Orange king”/C 0.1%; V23 - B. spectabilis ”Orange
king”/C 0.2%; V24 - B. spectabilis ”Orange king”/C 0.3%; V31 (Ct3) - B. buttiana
”Alba”/w; V32 - B. buttiana ”Alba”/C 0.1%;V33 - B. buttiana ”Alba”/C 0.2%; V34 - B.
buttiana ”Alba”/C 0.3%; V41 (Ct 4) - B. spectabilis/w; V42 -B. spectabilis /C 0.1%; V43
- B. spectabilis /C 0.2%; V44 - B. spectabilis /C 0.3%; V51 (Ct 5) - B.glabra/w;V52 - B.
glabra /C 0.1%;V53-B. glabra /C 0.2%;V54 - B. glabra /C 0.3%. The rooted cuttings
were planted in a 2:1 mixture of peat + perlite in pots of 18 cm (a size recommended in
the literature). Three foliar treatments with Cycocel were applied at a two-week
interval.
The observations and determinations were carried out may-october 2015
period and consisted on: the average plant height, the average shoots length, the
average internodes length, the average leaf length and the percent of blooming plants
under the influence of applied treatments.

RESULTS AND DISCUSSIONS

The growth retardants (PGRs) are commonly used in floriculture to reduce
plant growth, thus creating much more compact shrubs and beautiful foliage, but they
are also used for stimulating flowering (Mandă & Nicu 2016). The graph 1 shows that
the plants responded differently to Cycogan treatments for the studied species and
varieties. The average height of the plants treated with Cycogan in different
186
concentrations for B. buttiana "Raspberry Ice" and B. glabra, had lower values in
comparison to the untreated control plant. The retardant effect was stronger in B.
glabra, causing a significant reduction in the average plant height from 37.5 cm
(V51Mt) to 23.5 cm (V54). There was found that by applying treatments with different
concentrations of Cycogan in B.buttiana "Alba”, the effect was not to reduce the
vegetative growth. The values corresponding to the average plant height were superior
to the untreated control plant, ranging between 35 cm (V32) and 54 cm (V33) in
comparison to 34 cm (V31-Mt). There was found that by applying treatments with
different concentrations of Cycogan in B. spectabilis, the retardant effect was only seen
in the variants treated with Cycogan 0.2% (V43), and in B. spectabilis "Orange King" at
Cycogan 0.1% concentrations, respectively 0.2% (V22, V23). The growth was
stimulated at the plants treated with Cycogan 0.3%, obtaining higher values than in
comparison to the untreated control plant (34.8 cm - 40 cm, in comparison to 34 - 39
cm).
Regarding the average shoots length, the B. buttiana "Raspberry Ice" and B.
spectabilis "Orange King" recorded lower values than at the untreated control plant in
the case of concentrations of 0.1% (18 cm, respectively 21 cm) and 0.2% (16 cm - 22
cm). The retarding effect of Cycogan treatments was manifested in B. glabra at all
three concentrations, but the best results were obtained at the variants treated with
Cycogan 0.1% and 0.2% (V52, V53). In B. buttiana "Alba", the retarding effect was
shown only in the plants treated with Cycogan 0.1%, and in B. spectabilis the Cycogan
treatments acted as growth stimulators, as shown in figure 2. The treatments with
Cycogan 0,3% had a stimulating effect of vegetative growth in the treated variants
compared to the untreated control plants (23-43cm in comparison to 20-34cm).
From the analysis of the data presented in graph 2, the average internodes
length was considerably influenced by the application of growth retardants to all the
studied species. The application of the Cycogan treatments in B. spectabilis "Orange
King" determined the elongation of the internodes in all treated variants compared to
the untreated control plant. The average length of the internodes was between 1.8 cm
(V22) and 2.1 cm (V24), and the lowest value was recorded by V41Mt (1.6 cm). The
values of this parameter in B. spectabilis increase up to a concentration of 0.2%, then
decrease, and there is a reduction in the average length of the internodes at
concentrations of 0.3%. The values obtained at the end of the experimental period in
the B. buttiana "Alba", were superior to the control plant at concentrations of 0.2% and
0.3%. Only the concentration 0.1% determined a reduction in the length of the
internodes compared to the untreated control plant (from 1.8 cm to 1.5 cm). The
average length of the internode in the B. buttiana "Raspberry Ice" was higher in
comparison to the control plant at the concentration of 0.1% or equal to that of the
untreated control plant at the concentration 0.2%. There was a slight reduction in the
length of the internode only at the concentration of 0.3% in comparison to the
untreated control plant (from 0.9 cm to 0.8 cm). Inferior or equal values of the average
length of the internodes (0.8 cm and 1.1 cm, respectively) were obtained in B. glabra,
at the Cycogan-treated variants compared to the untreated control plant (1.1 cm).
The graph 3 shows that the average length of the leaves varied very little for
each studied species due to the Cycogan treatments. The application of the growth
retardant in B.buttiana Raspberry Ice and B. buttiana ”Alba” determined a decrease of
leaf length with increasing Cycogan concentration, from 6.3 cm (V11Mt) to 5.9 cm
(V14) for B. buttiana "Raspberry Ice" and from 6.9 cm (V31Mt) to 6.4 cm (V33) for B.
buttiana "Alba". Minimal values of the average leaf length were obtained in B. glabra at
the Cycogan 0.1% treated variant (V52 - 4.6 cm) in comparison to the untreated control
plant (V51-5.1 cm). The treatment with Cycogan 0.2% and 0.3% resulted in an
187
increase of the average leaf length. The average leaf length was superior in B.
spectabilis in comparison to the control plant, in the case of the concentration 0.1%,
and in B. spectabilis "Orange King" for the concentrations of 0.1% and 0.3%. The
concentration 0.2% determined a reduction of the average leaf length compared to the
untreated control plant (from 6.4 cm to 5.9 cm at B. spectabilis "Orange King" and from
8.1 cm to 7 cm at B. spectabilis). The only species with phytotoxicity (discoloured leaf)
was B. buttiana "Alba" (V34 - C 0.3%).
Regarding Bougainvillea flowering, the influence of Cycogan treatments are
analyzed for each species, knowing that the different Bougainvillea species/varieties
are blooming at a certain amount of light intensity. In the didactic greenhouse of the
Floriculture discipline, Bougainvillea is located within a sunny exposure with the
intensity of light ranging from 12,000-18,000 lux in May and September.
Under these circumstances, only B. glabra, B. spectabilis "Orange King" and
Bougainvillea Buttiana "Alba" flourished in October 2015. The flowering took place in
all experimental variants, in B. spectabilis "Orange King", B. glabra, the best results
(75%) were recorded by V23-0.2%, respectively V52-0.1% in terms of percentage of
flowering plants per variant. The flowering was observed in B. buttiana "Alba" at all the
treated variants, the best results on the percentage of flowering plants were recorded
by V33-0,2% (75%). The flowering was not influenced in B. spectabilis by Cycogan
treatments, the only flowering being observed in the untreated control plant (graph 4).

(cm) Lungime medie lăstar Înălțime medie plantă

60
50
40
30
20
10
0
V21 (Ct 2)

V31 (Ct 3)

V41 (Ct 4)

V51 (Ct 5)
V12-C 0.1%

V22-C 0.1%

V32-C 0.1%

V42-C 0.1%

V52-C 0.1%
V11 (Ct1)

V13- C 0.2%
V14- C 0.3%

V23- C 0.2%
V24- C 0.3%

V33- C 0.2%
V34- C 0.3%

V43- C 0.2%
V44- C 0.3%

V53- C 0.2%
V54- C 0.3%

B.buttiana ” Raspberry B.spectabilis ”Orange B.buttiana”Alba” B.spectabilis B.glabra

Ice” king”

Graph 1. The influence of Cycogan treatments on the plant growth

(cm)
2.5
2
1.5
1
0.5
0
V11 (Ct1)

V21 (Ct 2)

V31 (Ct 3)

V41 (Ct 4)

V51 (Ct 5)
V12-C 0.1%

V22-C 0.1%

V32-C 0.1%

V42-C 0.1%

V52-C 0.1%
V13- C 0.2%
V14- C 0.3%

V23- C 0.2%
V24- C 0.3%

V33- C 0.2%
V34- C 0.3%

V43- C 0.2%
V44- C 0.3%

V53- C 0.2%
V54- C 0.3%

B.buttiana ”Raspberry B.spectabilis ”Orange B.buttiana”Alba” B.spectabilis B.glabra

Ice” king”

Graph 2: The influence of Cycogan treatments on the average internodes length

188
 (cm)
10
8
6
4
2
0

V21 (Mt 2)

V31 (Mt 3)

V41 (Mt 4)

V51 (Mt 5)
V11 (Mt1)
V12-C 0.1%

V22-C 0.1%

V32-C 0.1%

V42-C 0.1%

V52-C 0.1%
V13- C 0.2%
V14- C 0.3%

V23- C 0.2%
V24- C 0.3%

V33- C 0.2%
V34- C 0.3%

V43- C 0.2%
V44- C 0.3%

V53- C 0.2%
V54- C 0.3%
B.buttiana”Raspberry B. spectabilis ”Orange B. buttiana”Alba” B. spectabilis B. glabra
Ice” king”

Graph 3: The influence of Cycogan treatments on the average leaf length

(%)
80

60

40

20

0
V11 (Ct1)

V21 (Ct 2)

V31 (Ct 3)

V41 (Ct 4)

V51 (Ct 5)
V33- C 0.2%

V53- C 0.2%
V12-C 0.1%
V13- C 0.2%
V14- C 0.3%

V22-C 0.1%
V23- C 0.2%
V24- C 0.3%

V32-C 0.1%

V34- C 0.3%

V42-C 0.1%
V43- C 0.2%
V44- C 0.3%

V52-C 0.1%

V54- C 0.3%
B.buttiana ” Raspberry B.spectabilis ”Orange B.buttiana”Alba” B.spectabilis B.glabra
Ice” king”

Graph 4: The influence of Cycogan treatments on the percent of blooming plants

CONCLUSIONS
All the analyzed growth parameters in the Bougainvillea species were
influenced differently for each species, with no determination for an optimal
concentration at which Cycogan could have a retardant effect for all the studied
species / varieties. The best results in terms of the retardant effect of Cycogan on the
plants were obtained in: B. buttiana "Raspberry Ice" - Cycogan 0.1% and 0.2%; B.
spectabilis "Orange King" - Cycogan 0.1% and 0.2%; B. buttiana "Alba" - Cycogan
0.1%; B. spectabilis - Cycogan - 0.2%; B. glabra - Cycogan 0.2% and 0.3%. The only
species with phytotoxicity (discoloured leaf) was B. buttiana "Alba" (V34 - Cycogan
0.3%). In terms of flowering, only B. glabra, B. spectabilis "Orange King" and B.
buttiana "Alba" flourished in October 2015. The B. spectabilis flowering was not
influenced by Cycogan treatments, the only flowering being observed at the untreated
control plant.

REFERENCES
Aldrich, J. H., & Norcini, J. G., 1996, Effect of Plant Growth Regulators on Growth
of'Barbara Karst'Bougainvillea. In Proceedings-Florida State Horticultural Society (Vol.
109, Pp. 3-4). Florida State Horticultural Society.
Cirillo Chiara, Youssef R, R. Caputo, G. Raimondi and St. De Pascale, 2014, The
Influence of Deficit Irrigation on Growth, Ornamental Quality, and Water Use Efficiency
of Three Potted Bougainvillea Genotypes Grown in Two Shapes. HortScience October
2014 vol. 49no. 10 1284-1291.
189
 Fang-Yin LIU and Yu-Sen CHANG, 2011, Ethephon treatment promotes flower
formation in bougainvillea. Botanical Studies (2011) 52: 183-189.
Hackett, W.P., R.M. Sachs, 1967, Chemical control of flow-ering in Bougainvilllea
'San Diego Red'. Proc. Amer. Soc. Hortic. Sci. 90: 361-364..
Jain R., T, Janakiram K. S., Kumawat G., 2014, Induction of dwarfing in
bougainvillea cv Mahara by use of growth regulators. Indian Journal of Agricultural
Sciences, 84(7), 802-7.
Jain, R., Janakiram, T., Kumawat, G., 2016, Effect of growth retardants on growth
and flowering of bougainvillea (Bougainvillea spectabilis) cv. Shubra. The Indian
Journal of Agricultural Sciences, 86(9).
Kobayashi D.K., 2007, Bougainvillea. Departament of Tropical Plant and Soil
Sciences, University of Hawaii at Manoa.
Mandă M., Carmen Nicu, 2016, The influence of growth retardants and substrate
volume on growth of Hemigraphys repanda L. Analele Universitatii din Craiova, Seria
Biologie, Horticultura, TPPA, Ingineria Mediului, Vol. XVII (LIII): 263-268.
Probert M., Leela Hussain, A.A. Awan, 1999, Probert Encyclopaedia –
Inflorescence , Southampton, United Kingdom.
Randhawa G.S., A. Mukhopadhyay, 1986, “Floriculture in India”, Allied Publishers
Ptv. Limited.
Shao Z., Yang Y., Qiu S., 2006, The study of advance of flowering time regulator
of Bougainvillea glabra. Chinese Agriultural Science Bulletin 22: 326–9.
Saifuddin M., A.B.M.S. Hossain, O. Normaniza, A. Nasrulhaq Boyce and K.M.
Moneruzzaman, 2009b, The effects of naphthaleneacetic acid and gibberellic acid in
prolonging bract longevity and delaying discoloration of Bougainvillea spectabilis.
Biotechnology, 8(3), 343-350.
Whistler W.A., 2000, Tropical ornamentals: A guide.Timber Press, Portland.

190
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ANNALS OF T HE  Horticu lture
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EFFECTS OF TEMPERATURE ON GROWTH RATE, LIPID AND PROTEIN

CONTENT OF THE MICROALGAL SPECIES Nannochloropsis oculata

M. N. Metsoviti1*, N. Katsoulas1, I. T. Karapanagiotidis2, G. Papapolymerou3

1
Department of Agriculture Crop Production and Rural Environment, University of Thessaly, Volos, Greece
2
Department of Ichthyology and Aquatic Environment, University of Thessaly, Volos, Greece
3
Department of Civil Engineering, Technological & Educational Institute of Thessaly, Larissa, Greece
* Correspondence author. [email protected]

Key words: microalgae, Nannochloropsis oculata, productivity, growth rate, nutrient composition

ABSTRACT
This paper presents the effects of three different temperatures (20, 25 and 30 °C) on
the growth rate, productivity and nutrient composition of the microalgal species Nannochloropsis
oculata. The experiments were conducted in closed photobioreactors of 25 L capacity, where the
temperature and the light intensity were controlled. The absorbance at 665 nm was measured
daily. Total productivity was determined at the end of the experiments and the specific growth
rates in exponential growth phase (μexp) were calculated. Additionally, the nutrient composition of
the algal biomass in lipids and proteins was determined. It was found that N. oculata grew best
at 30oC. The protein content was similar among the three different temperatures, while the lipid
content increased with the increase of temperature.

INTRODUCTION
Microalgae are unicellular photosynthetic organisms that use light and carbon
dioxide, with higher photosynthetic efficiency than plants, for the production of
biomass. As aquatic relatives of plants, they thrive in aerated, liquid cultures where the
cells have sufficient access to light, carbon dioxide and other nutrients (Rosenberg et
al. 2008). Their biodiversity is enormous. There are species that prosper in freshwater
and others in saline water, such as the marine microalgal species Nannochloropsis
oculata.
Microalgae can be cultivated in open or closed systems. Open-field cultivation
is usually conducted in an open pond or an open container that is exposed to the
environment. Cultivation in a closed system can be carried out in photobioreactors. In
closed systems, the productivity of biomass is higher than it is in the open systems and
the risk of contamination of the cultivation is lower. However, investment and operating
costs, as well as energy requirements, are higher than in open systems (Rösch &
Posten 2012).
The microalgae culture systems are quite complex and are influenced by
different factors, such as temperature, carbon dioxide, pH, light intensity, mixing grade,
salinity, as well as the nutrient composition of the culture medium (nitrogen, potassium,
phosphorous and micronutrients) (Dean et al. 2010).
Cultivation in stress conditions includes culture in inadequate nutrients in the
culture medium, very high or low light intensities and low pH. These conditions lead to

191
a reduction in the rate of growth and as a result to a decrease in productivity (Wijffels
et al. 2010).
Environmental and culturing condition variations (temperature, light intensity,
nutrient content of the culture medium) can alter the chemical composition of the
biomass of microalgae produced at the end of the cultivation (Gouveia & Oliveira
2009). For instance, as the nitrogen concentration increases in the culture medium, the
protein content increases and the lipid content decreases (Metsoviti et al. 2016). It
should be mentioned that, the lipid production of a microalgal species can be
enhanced, by cultivation in different temperatures (Sibi et al. 2016).
Microalgae are used in pharmaceutical, in biodiesel production, in
biodegradation, as nutritional supplements and as feed for animals and fish, replacing
other ingredients of the feeds (Pulz & Gross 2004). In aquaculture, various species of
microalgae are used for fish larval nutrition during a brief period either for direct
consumption in the case of molluscs and peneid shrimp or indirectly as food for the live
prey fed to fish larvae (Muller-Feuga 2000).
The present study was conducted in order to investigate the effects of three
different temperatures on the growth rate, total productivity and nutrient composition in
lipids and proteins of the marine microalgal species Nannochloropsis oculata. For this
purpose, N. oculata was cultivated in a closed photobioreactor at three different
temperatures, 20, 25 and 30 oC in the same culture medium and with the same light
intensity. The total production on a dry basis (g of dry algal biomass/ 25 L of culture
medium), the specific growth rate in the exponential growth phase (μexp) and the
maximum absorbance readings (αmax) for the three different temperatures, were
determined as well.

MATERIAL AND METHODS

Microalgal culture: The microalgal species N. oculata was obtained from the
Experimental Phycology and Culture Collection of Algae from the University of
Goettingen in Germany (EPSAG).
Cultivation system: All experiments were conducted in a closed
photobioreactor of 25 litres capacity (Figure 1), at three different temperatures, 20, 25,
30 oC and were illuminated with red and white led lamps (each, 120 μmol m −2 s−1) on
12:12 h light-dark cycles. The temperature was controlled to within ± 0.1oC. The
duration of each experiment was 25 days.
Microalgal culture media: The culture medium was the same in all experiments
and contained the following ingredients, suggested by Andersen et al. 1991: Each litre
of the culture medium contained: 0.2g KNO3/L, 0.02 g K2HPO4/L, 0.02 g
MgSO4.7H2O/L, 30 ml of soil extract/L, 5 μg vitamin B12/L and 5 ml/L, of a solution
containing the following micronutrients: (1 mg ZnSO 4.7H2O, 2 mg MnSO4.4H2O, 10 mg
H3BO3, 1 mg Co(NO3)2.6H2O, 1 mg MoO4.2H2O, 0.005 mg CuSO4.5H2O, 700 mg
FeSO4.7H2O and 800 mg EDTA)/L. Additionally, synthetic salt was added in the culture
medium so that the salinity of the water was maintained at 35 ‰.
MIcroalgal biomass concentration: The microalgal concentration was
determined daily by optical density measurements at 655nm with the use of a
spectroscopy UV/Vis instrument. Three samples were collected daily and all measures
were carried out in triplicate. Additionally, at the end of the experiment total productivity
of each culture was determined (in g/25 L), after collection of the biomass by a
condensation process, which was based on flocculation, natural sedimentation,
removal of the supernatant, collection of the condensates and removal of moisture and
the specific growth rates in the exponential growth phase (μexp) were calculated
according to the relation:
192
 μ exp = ln (α2/α1) / (t2-t1)

where, α1 and α2 are the absorbances at the beginning and the end of exponential
growth phase at time 1 (t1) and 2 (t2), respectively.

Figure 1. Closed photobioreactor, where the experiments were conducted.

Protein and lipid content: Crude protein of the microalgal biomass was
determined by Kjeldahl method, using a conversion factor of 6.25 (AOAC 1997). Lipids
were extracted from the samples with 1:1 chloroform/methanol with Folch et al. (1957)
method. According to Ryckebosch et al. 2012, chloroform–methanol 1:1 was shown to
be the best solvent mixture for extraction of total lipids from microalgae.
Statistical analysis: Results are expressed as mean ± standard deviation. Data
were analyzed using a one-way analysis of variance (ANOVA). The significant
differences between treatments (P<0.05) were determined using Tukey's multiple
comparison test.

RESULTS AND DISCUSSIONS

Figure 2 illustrates the absorbance readings versus the cultivation time of N.
oculata during the cultivation at three different temperatures (20, 25 and 30 oC). Table
1 presents the specific growth rates in exponential growth phase (μexp), the maximum
absorbance readings (αmax) and total biomass productivities on a dry basis (in g/25L)
for N. oculata grown at three different temperatures while, Table 2 shows the crude
protein and crude lipid contents (%) of N. oculata cultivated at the three different
temperatures.
It was found that the growth rate of N. oculata (Fig. 2 and Table 1), was
strongly dependant on the cultivation temperature. The increase in temperature from
20oC to 25oC and to 30oC, led to consequent increases in specific growth rates in the
exponential growth phase (μexp), in the maximum absorbance readings (αmax) and in
total productivities.

193
 Nannochloropsis oculata

0,9
30oC
0,8

0,7

0,6 25oC
Absorbance

0,5

0,4

0,3

0,2 20oC
0,1

0
0 5 10 15 20 25 30
Time (in Days)

Figure 2. Growth curves of the microalgal species N. oculata grown at three different
temperatures.

Specifically, the total productivity in algal biomass was doubled at 30oC,

o
compared with the productivity at 20 C. Thus, it appears that the cultivation of N.
o
oculata is favored at 30 C, compared to lower temperature regimes.

TABLE 1
Specific growth rates in exponential growth phase (μexp), maximum absorbance
readings (αmax) and total productivities (g/25 L) for N. oculata grown at three different
temperatures.

T→ 20oC 25oC 30oC

μexp 0.24 0.28 0.35
αmax 0.19 0.59 0.82
Productivity (g/25 L) 30.2 52.3 60.3
TABLE 2
Lipid and protein content (% of dry weight) of N. oculata grown at three different
temperatures. Values represent means ± st. deviation (n=3). Values in the same row
bearing different superscript indicate statistically significant difference (P < 0.05).

Nutrients 20oC 25oC 30oC

a b c
Lipids (%) 9.0 ± 0.2 13.2 ± 0.5 15.7 ± 0.4
Proteins (%) 16.9 ± 0.3a 17.4 ± 0.4a 17.1 ± 0.1a

194
 As far as the nutrient composition of N. oculata is concerned, the protein
content of the algal biomass remained constant at the three different temperatures
(Table 2). This may be due to the fact that the protein is a significant structural and
metabolic component of algal cells, so that their protein content might be more
resistant to temperature alterations.
On the other hand, the lipid content of the algal biomass increased significantly
with the increase of the temperature from 20oC to 25oC and to 30oC. These findings
are in agreement with those reported by Converti et al. (2009), in which the lipid
content of N. oculata was doubled when cultured at 25oC compared to the temperature
of 20oC. The increase in lipid content by rises in temperature has also been shown for
other microalgal species such as S. obliquus (Vitova et al. 2015).
Total lipid content in microalgae increases to a certain extent as the
temperature increases and reaches an optimal level (Sibi et al. 2016). The optimal
value of temperature where the highest biomass and lipid production is achieved varies
from species to species and as a result, it is difficult to generalize on specific influence
of the environmental factors on growth, biochemical composition and enzyme activities
in different microalgal species (Gigova et al. 2012). Unfortunately, a higher
temperature regime that could have resulted in a possible peak in lipid content was not
tested in the present study.
It should be stated, however, that total lipid content does not signify lipid
quality and that culture of microalgae at higher temperatures may also alter their fatty
acid compositions. It is known, for example, that higher temperatures decrease the
production of unsaturated lipids in Dunaliella salina (Thompson, 1996). Therefore,
future research on the effects of temperature on the lipid content of microalgae should
also consider alterations in lipid quality, especially if their use is intended as nutrient
supplements.

CONCLUSIONS
o
Growth of the microalgal species N. oculata at 30 C was found to be the best
temperature regime in terms of growth rate, absorbance and productivity compared to
20oC and 25oC. Total lipid content of N. oculata was increased concomitantly with
increases in temperature, but protein content was unaffected.

REFERENCES
AOAC, 1997, Official Methods of Analysis, 16th ed. Association of Official
Analytical Chemists, Washington, DC.
Andersen R. A., Jacobson D. M., Sexton J. P., 1991, Provasoli-Guillard center for
culture of marine phytoplankton catalogue of strains. West Boothbay Harbor, Maine,
USA, pp. 98.
Converti A., Casazza A. A., Ortiz E. Y., Perego P., Del Borghi M., 2009, Effect of
temperature and nitrogen concentration on the growth and lipid content of
Nannochloropsis oculata and Chlorella vulgaris for biodiesel production. Chemical
Engineering and Processing: Process Intensification. 48(6):1146–1151.
Dean A., Sigee D., Estrada B., Pittman J., 2010, Using FTIR spectroscopy for
rapid determination of lipid accumulation in response to nitrogen limitation in
freshwater microalgae. Bioresour. Technol. 101, 4499–4507.
Folch J., Lees M., Sloane-Stanley G. H., 1957, A simple method for the isolation
and purification of total lipides from animal tissues. J Biol Chem1957, 226:497-509.
Gigova L., Ivanova N., Gacheva G., Andreeva R., Furnadzhieva S., 2012,
Response of Trachydiscus minutus (Xanthophyceae) to temperature and light. J.
Phycol. 48, 85–93 (2012).
195
 Gouveia L., Oliveira A. C., 2009, Microalgae as a raw material for biofuels
production. Journal of Industrial Microbiology & Biotechnology 36(2), 269-274.
Metsoviti M. N., Papapolymerou G., Karapanagiotidis I. T., Katsoulas N., 2016,
Effect of different concentrations of nitrogen on the growth rate and nutrient content of
the microalga Chlorella vulgaris. 2nd International Congress on Applied Ichthyology &
Aquatic Environment 10.p.72-76.
Muller-Feuga A., 2000, The role of microalgae in aquaculture: situation and trends.
J. Appl. Phycol. 12:527–534.
Pulz O., Gross W., 2004, Valuable products from biotechnology of microalgae.
Applied Microbiology and Biotechnology. 65 (6), 635–648.
Rösch C., Posten C., 2012, Challenges and Perspectives of Microalgae
Production, Karlsruhe, Karlsruhe Institute of Technology (KIT), 5.
Rosenberg J. N., Oyler G. A., Wilkinson L, Betenbaugh M.J., 2008, A green light
for engineered algae: redirecting metabolism to fuel a biotechnology revolution. Curr
Opin Biotechnol 19:430–436.
Ryckebosch Ε., Muylaert Κ., Foubert Ι., 2012, Optimization of an Analytical
Procedure for Extraction of Lipids from Microalgae. J Am Oil Chem Soc 89:189–198.
Sibi G., Shetty V., Mokashi K., 2016, Enhanced lipid productivity approaches in
microalgae as an alternate for fossil fuels - a review. Journal of the Energy Institute.;
89 (3):330–334.
Thompson, G.A., 1996, Lipids and membrane function in green algae. Biochim.
Biophys. Acta, 1302, 17–45.
Vitova M., Bisova K., Kawano S., Zachleder V., 2015, Accumulation of energy
reserves in algae: from cell cycles to biotechnological applications. Biotechnology
Advances. 33(6):1204–1218.
Wijffels R. H., Barbosa M. J., Eppink M. H. M., 2010, Microalgae for the production
of bulk chemicals and biofuels. Wiley InterScience. Biofuels, Bioprod. Bioref. 4:287–
295.

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ANNALS OF T HE  Horticu lture
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Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

AMPELOGRAPHIC CHARACTERISTICS OF CHARDONNAY 76

1 1 1*
Nakov Zdravko , Ivanov Miroslav , Simeonov Iliyan

* Institute of Viticulture and Enology, Pleven, Bulgaria, e-mail: [email protected]

Key words: vine, variety, clone, ampelographic description

ABSTRACT
During the period 2008-2011, an ampelographic description of the introduced French
clone Chardonnay 76 was made at the Institute of Viticulture and Enology – Pleven. The study
found that Chardonnay clone 76 belonged to the group of medium-ripening white wine varieties.
In the soil and weather conditions of the region of Pleven, its grapes reach technological maturity
during the first ten days of September (07.09.). The cluster is medium sized, cylindrical-conical in
shape, with one small wing, semi-compact or compact. The berry is small to medium-sized,
yellow-greenish, almost spherical. The vines have medium to strong growth, good fertility and
medium yield. The clone is not liable to putting forth catkins and milerandage. It has good
resistance to low winter temperatures. It is sensitive to fungal diseases and especially to
powdery mildew (oidium). It develops well and gives yield grafted to Berlandieri x Riparia SO4
rootstock. Chardonnay 76 has a very good sugar accumulating capacity - 23.20% sugars while
3
keeping enough titratable acids - 7.09 g/dm . Chardonnay 76 wine is yellow-greenish in colour
with a pleasant floral fruity aroma, pronounced acidity, dense, harmonious and long-lasting
aftertaste.

INTRODUCTION
The origin of Chardonnay variety has not been fully clarified however
according to Viala and Vermorel (1905) and Galet (1958; 1976), it was a French variety
grown since the old age in the regions of Burgundy and Champagne. The variety
belongs to the Western ecological and geographical group. Quality white dry wines and
products for sparkling wines are made from its grapes. Because of its valuable
economic qualities, Chardonnay variety is widely distributed in most wine-growing
countries in Europe, North and South America, South Africa and Australia. In Bulgaria
the variety was introduced from France, studied by Nedelchev and Kondarev (1936),
and later it was zoned to be distributed (Stoev, 1960).
During its many years of development and growing under different soil and
climatic conditions, various mutational changes occurred in the population of
Chardonnay variety. Most of them were of a negative nature, reducing the yield and
deteriorating grapes quality. That necessitated the study of the intra-diversity variety
and the application of the clonal selection method to improve some of the significant
economic qualities of the variety. Many clones were obtained by the clonal selection
method in Italy (Moretti, 1994), France (Roychev, 2012; Galet, 1994), Russia
(Guseynov et al., 2011), Germany and other vine-growing countries. In Bulgaria, a
clone of Chardonnay 6/48 (Nakov, 2006) was selected and approved in 1994. For
enriching the grapevine gene pool of the country and diversifying the range of wines, a

197
French clone of Chardonnay variety was introduced at the Institute of Viticulture and
Enology in Pleven in 1991.
The objective of this study is to present detailed ampelographic characteristics
of Chardonnay Clone 76 introduced from France.

MATERIAL AND METHODS

Chardonnay 76 was selected in France by the method of clonal selection. The
ampelographic study of the variety was carried out during the period 2008 – 2011 in
the clonal section established for that purpose in the Experimental Base of the Institute
of Viticulture and Enology – Pleven. The vines were grafted to Berlandieri x Riparia
SO4 rootstock and were planted at a distance of 3.00/1.30 m. They were grown on
Moser training system with stem height of 0.80 m. The vines were mixed pruned with
equal individual loading - 32 winter eyes (6 х 2 eyes and 2 х 10 eyes).
The botanic description (young shoot, mature shoot, developed leaf,
inflorescence, cluster and berry), the phonological observations (onset of flowering,
onset of grapes ripening and technological maturity), the fertility indicators (ratio of
developed winter eyes and fruit shoots with 1, 2, 3 clusters, ratio of developed and fruit
shoot) and yield (average mass per cluster and average yield per vine) was done in
accordance with the approved methodology in the country for clonal selection (Katerov
et al., 1990).
During the grapes ripening phase, the dynamics of sugar accumulation was
monitored. At technological maturity the quality parameters (sugars and titratable
acids) were determined and a mechanical analysis was made to establish the texture
and the structure of the cluster and the berry. From each harvest at the Experimental
cellar of IVE - Pleven, 30 kilograms were processed according to the classical
technology for the production of white dry wines by the method of micro-vinification
(Yankov, 1992). The young wines were subjected to a chemical analysis according to
the common methods used in the wine industry (Ivanov, 1979) and an organoleptic
evaluation by the 100-score system was made.

RESULTS AND DISCUSSIONS

ORIGIN. Chardonnay Clone 76 was separated from Chardonnay varieties by
the method of clonal selection in France.
DISTRIBUTION. In Bulgaria Chardonnay 76 is distributed in the super-elite
mother plantation of IVE – Pleven.
BOTANIC DESCRIPTION
Young shoot. The crown is very mossy to cottony, with a pinkish border. The
first leaf is gray-green, quite mossy on the top side, and cottony on the underside. The
second leaf is waxy-yellowish, mossy on the top side and intensely mossy on the
underside. The rest of the leaves are golden-yellowish, with a bronze tinge, mossy on
the top side and intensely mossy with bristles on the underside. The young shoot is
gray-greenish, with wine-red strips, slightly mossy.
Mature shoot. The internodes are medium long, medium thick, slightly
flattened, slightly angular. The bark is light brown. The nodes are darker in color. The
mature shoot has a small core and solid wood.
Developed leaf. The leaf blade is medium large, wider than longer, three to
five-lobed, folded upward. The top side is slightly reticulated wrinkled, with arachnoidal
moss, while the bottom surface - slightly mossy-bristly. The outlines of the middle
section form an obtuse angle. The upper lateral notches are shallow, open, in the form
of an acute angle, and rarely moderately deep, lyrated. The bottom side notches are
shallow, barely noticeable, open, in the form of an acute angle or missing. The tail
198
notch is open, lyrated or arched, equilateral. The top teeth are small to medium-sized,
straight, triangular, while the end ones are small, transient-domed. The veins are
green, on top - with scarce bristles and underneath - mossy. The petiole is short,
green, somewhat pale wine-red, slightly mossy-bristly. The autumn colouring of the
petiole is yellow.
Inflorescence. It is monoecious, with 5 stamens, rarely 4 or 6 stamens.
Cluster. It is medium large (13.3/8.1 cm), cylindrical-conical, with one small
wing, semi-compact or compact. The rachis is short, medium thick, woody at the base.
the average mass per cluster is 125.0 g (Fig. 1).

Fig. 1. Cluster of Chardonnay 76

Berry. It is small to medium large (13.61/13.60 mm), almost spherical,

sometimes deformed because of the cluster compactness. The skin is yellowish-green,
thin and elastic, with slight wax coating. The texture is juicy and the taste –
harmonious, neutral. The average mass of 100 berries is 146.7 g.

AGROBIOLOGICAL CHARACTERISTIC
Vegetation period. Chardonnay 76 is a medium ripening white wine variety.
Under the climatic and soil conditions of the region of Pleven, its grapes reach
technological maturity in the first ten days of September (07.09) and depending on the
weather conditions of the year it varies from the end of August (26.08.) to mid
September - (16.09.). The vegetation period duration is about 145 days. Flowering
starts in the last ten days of May - 21.05. and ranges from 17.05. to 24.05. The onset
of grapes ripening occurs at the end of July (29.07.) and varies from 21.07. to 04.08.
depending on the weather conditions of the year (Table 1).

Table 1
Phenological observations in Chardonnay 76 from 1998-2011 periode

Phenophases Data
Beginning of flowering 21.05.
Beginning of ripening of the grapes 29.07.
Technological ripeness in grapes 07.09.

199
 Growth strength. Vines have medium intense growth. By the end of the
vegetation the shoots mature well.
Fertility and yield. Chardonnay 76 is distinguished with good actual fertility –
84.12 % (80.00 % – 88.00 %) developed winter eyes and 79.47 % (67.31 % – 89.74
%) fruit shoots. From the fruit shoots 37.50 % (31.59 % – 43.75 %) have 1 cluster;
56.60 % (50.00 % – 62.86 %) - 2 clusters and 5.90 % (0.00 % - 10.23 %) – 3 clusters.
The fertility ratio per developed shoot is 1.28 (0.96 – 1.46) and of a fruit shoot – 1.61
(1.43 – 1.69). The average yield per vine is 3.086 kg, varying from 2.264 kg to 3.710 kg
(Table 2).
Table 2
Actual fertility indexes of Chardonnay 76 from the 1998-2011 periode

Fruiting shoots: Fertility coefficient in:

Developed
with 1 with 2 with 3
buds total developed
cluster clusters clusters fruit shoot
shoot
% % % % %
84.12 79.47 37.50 56.60 5.90 1.28 1.61

Putting forth catkins and milerandage. The clone is distinguished by good

pollination and fertilization and it is not liable to putting forth catkins and milerandage.
Disease resistance. It is not resistant to fungal diseases and especially to
powdery mildew. In years of rainy autumn grapes are attacked strongly by gray rot.
Agrotechnical specifics. Chardonnay 76 requires mixed pruning with
individual loading per vine of 32 winter eyes (6 spurs х 2 eyes and 2 canes х 10 eyes).
When grown on Moser training system and planting density 256 vines/dka (3.00 х 1.30
m.), the clone gives optimum yield with very good grape quality.
Suitable rootstoks. Chardonnay 76 has good affinity to Shasla x Berlandieri B
and Berlandieri x Riparia SO4 rootstocks.
Response to environmental conditions. The clone has good resistance to
low winter temperatures and recovery capacity, allowing it to be grown on various stem
training systems in all vine-growing regions of the country.

TECHNOLOGICAL CHARACTERISTICS
Grapes mechanical composition. According to the cluster mechanical
composition Chardonnay 76 is a typical wine variety, with high content of solid fraction.
The rachis ratio in the cluster is 4.16 % (3.26 % - 4.96 %) and of the berries - 95.84 %
(96.74 % - 95.04 %). The skins in the berry are 9.68% (7.72 % - 11.50 %), the seeds -
4.24 % (3.26 % - 5.67 %) and the mesocarp - 86.08 % (82.83 % - 88.49 %). The
theoretical yield is 82.50 %, varying for the individual vintages from 79.03 % to 85.61 %
(Table 3).
Chemical composition. Chardonnay 76 is characterized by very good sugar-
accumulating capacity – 23.20 % sugars that vary depending on the weather
conditions from 22.40 % to 23.90 %. The titratable acids of the grapes are kept
relatively high - 7.09 g/dm3 (6.15 g/dm3 - 8.25 g/dm3). Chardonnay 76 wine is
yellowish-green in colour with a pleasant floral and fruity aroma, pronounced acidity,
dense, harmonious and long-lasting aftertaste (Table 3).
GENERAL ASSESSMENT OF THE CLONE
Chardonnay 76 is medium ripening white wine clone. It was selected in France
by the method of clonal selection from the population of Chardonnay variety. The
cluster is medium large, cylindrical-conical, with one small wing, semi-compact or

200
compact. The berry is small to medium large, almost spherical, sometimes deformed
because of the cluster compactness.
Table 3
Yield, mechanical and chemical analyses of grapes and wine of Chardonnay
76 from the 1998-2011 periode

Cluster mechanical composition

Weight, g 125.00
Length, cm 13.30
Width, cm 8.10
Rachis, % 4.16
Berries, % 95.84
Berry mechanical composition
Weight of 100 berries, g 146.70
Length, mm 13.61
Width, mm 13.60
Skins, % 9.68
Seeds, % 4.24
Flesh, % 86.08
Yield and chemical analyses of grapes
Average yield per vine, kg 3.086
Theoretical yield, % 82.50
Sugars, % 23.20
Titratable acids, g/dm 3 7.090
Chemical composition and tasting score of white wine
from Chardonnay 76
Alcohol, vol. % 12.82
3
Sugars, g/dm 1.60
Titratable acids, g/dm 3 6.84
pH 3.16
Sugar free extract, g/dm 3 22.78
Total phenolic compounds (TPC), g/dm 3 0.45
Tasting score 77.80

The skin is yellowish-green, thin and elastic, with slight wax coating. The
texture is juicy and the taste – harmonious, neutral. Vines have medium to intense
growth and average high fertility. It is not liable to putting forth catkins and
milerandage. It is not resistant to fungal diseases and especially to powdery mildew. In
years of rainy autumn grapes are attacked strongly by gray rot. The vines have good
resistance to low winter temperatures and recovery capacity. Chardonnay 76 has good
affinity to Berlandieri x Riparia SO4 rootstocks. When grafted to this it is obtained
medium high yield with very good grapes quality. Under the soil and weather
conditions of Pleven region Chardonnay 76 has very good sugar-accumulating
capacity by keeping relatively high titratable acidity. The grapes are suitable for the
production of quality white dry wines. Chardonnay 76 wine is yellowish-green in colour
with a pleasant floral and fruity aroma, pronounced acidity, dense, harmonious and
long-lasting aftertaste.

201
 REFERENCES
Galet P., 1958. Sepages et vignobles de Frans. Vol. II, Montpellier, France.
Galet P. 1976. Precis d‟ampelographie pratique. Monpellier, France, Paul Dehan,
260 p.
Galet P., 1994. Le Chardonnay. Revue des Oenologues, № 73 (9), 19-20.
Guseynov Sh., B. Chigrik, N. Gordeeva, 2011. Comparative evaluation of the
efficacy of various clones of the Chardonnay variety under Taman conditions.
Collection from the International Scientific and Practical Conference, Novocherkask,
57-60.
Ivanov T., S. Gerov, A. Yankov, G. Bambalov, T. Tonchev, D. Nachkov, M.
Marinov, 1979. Practice in Wine Technology. Plovdiv, ed. "Hristo G. Danov", pp. 530.
Katerov К., А. Donchev, М. Kondarev, G. Getov, Т. Nachev, Е. Hershhovich, V.
Valchev, М. Markova, D. Braykov, H. Todorov, P. Мамаrоv, Y. Ivanov, Z. Zankov, B.
Tsankov, L. Radulov, М. Ivanov, М. Jekova, 1990. Bulgarian Ampelographia, General
ampelographia, Publisher of BAS, V. І, 296 p.
Moretti G., 1994. Vitigni et cloni in Italia. Vitignevini, № 12, 13-53.
Nakov Z., 2006. Chardonnay clone 6/48. Viticulture and Enology, 5, 12-14.
Nedelchev N., М. Kondarev, 1936. Results of the cultivation of French varieties of
vines in Saranyvo. Yearbook Agr. Faculty of Forestry, vol ХIV, Sofia.
Roychev V., 2012. Ampelographia. Academic publisher of the Agricultural
University of Plovdiv, 574 с.
Stoev К., 1960. Regionalization of viticulture in Bulgaria, Sofia, Zemizdat, 18-22,
117-132.
Viala P., V. Vermorel, 1905. Ampelografie. Vol. 6, Paris, Masson, 476 p.
Yankov А., 1992. Winemaking Technology. Sofia, Zemizdat.

202
 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
 Food produce process ing
UNIVERSITY OF CRAIO VA
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

RESEARCH ON THE BEHAVIOR OF SOME HYBRIDS LETTUCE IN

SOLARIUM

Neață Gabriela1, Hoza Gheorghița1*, Dobrescu Aurelia1, Basarabă Adrian1,Dinu Maria 2

1 1
Mohammed Jasin Mohammed , Hadi Hoobi Shallat
1
University of Agronomical Sciences and Veterinary Medicine Bucharest, Horticulture Department,
Bd.Mărăști, 59, Bucharest, Romania
2
University of Craiova, Horticulture Department, Craiova, Romania
* Correspondence author: , e-mail:[email protected]

Key words: Lactuca sativa L., hybrids, growth regulator

ABSTRACT
The research was conducted in the solarium of the Department of Horticulture at
USAMV Bucharest in 2017. Six F1 hybrids of Lactuca sativa L were used: Touareg, Alanis,
Analena, Shangore, Centore and Sotalis. The behavior of lettuce was followed when applying
gibberelinic acid (AG3) at different doses during vegetation. Phenological observations,
biometric measurements, agrochemical determinations of lettuce quality, and production were
recorded. The results were statistically interpreted.

INTRODUCTION
Achieving a high level of agricultural crop production implies the combined use
of classical chemical or organic fertilizers as well as growth regulators that ensure
optimal supply of plants with nutrients.( Pavlou et al.2007).
Cultivation of crops is done under conditions of maximum economic efficiency
without producing negative effects on the quantity, commercial quality and the food
value of the production (http: // www.giberelina.ro).
Foliar fertilizers apply to most crop plants to provide complete and balanced
fertilization, stimulate rooting, growth and fructification by accelerating the absorption of
nutrients in the soil.( Tamme et al., 2009).
The discovery of stimulating growth regulators (gibberellins) has resulted in
spectacular effects with outstanding results in agricultural and horticultural production.
()The main changes that occur in the metabolism of plants under the action of
gibberellins are intensification of sweating and increased water consumption,
respectively faster absorption of nutrients and delaying aging processes of vegetal
tissues.
The purpose of the research was to track the lettuce's behavior when applying
gibberellin acid (AG3) at different doses during vegetation.

MATERIAL AND METHODS

The experience was carried out in 2017 in a USAMVB solarium, the
Department of Vegetables.

203
 The biological material used was represented by 6 F1lettuce hybrids: Touareg,
Alanis, Analena, Shangore, Centore and Sotalis.(Table 1).

Table 1.
Characterization of cultivated lettuce hybrids
No Hybrid Characteristics Producer
1 TOUAREG It has resistance to the attack of aphids, LMV (the Seminis
virus of the mosaic virus) as well as the hand. The
bright, bright green caps are compact and can reach
great size. The Touareg has a good tolerance to the
floral stems and to the marginal necrosis of the
leaves. Leaves of equal, fine, color and shape of the
captain make the Touareg an excellent variety for
marketing and fresh consumption. Touareg is
recommended both for open-field cultivation and
protected areas.
2 ALANIS For crops in the field or tunnels and solariums, Seminis
throughout the year, the captains are large, well-
closed, compact, light green, shiny. Alanis has
shown good tolerance to plant disease (Botrytis,
Sclerotinia) and good tolerance to floral stems,
resistance to LVW (Lettuce mosaic virus) and to
Bremialactucae, I-25
3 SHANGORE Butterhead autumn lettuce for late-winter autumn Syngenta
crops. The caps are compacting, semi-finished. The
base of the captains is closed, smooth and shining. It
is noted for its high resistance to Bremialactucae BI
1-24. The plants grow rapidly, the leaves are erect.
Forms large captains (400-600g) even on weak
soils. Easily cut off with erect leaves
4 ANALENA Analena is a large lettuce for unprotected culture, set Syngenta
up early in spring and autumn. The leaves are large,
of medium thickness and form large caps in a very
short time. The light green color of the leaves makes
this variety a very attractive one. Presents high mold
resistance (Bremia).
5 CENTORE The best choice for spring late winter. Shows closed Enza
and compact captains, high hand strength (HR) Bl 1- Zaden
24. It is not pretentious to soil and low temperatures.
The crop cycle is fast, it is resistant to drying the
inner leaves. The base of the captains is smooth and
well closed. The internal structure of the head is
compact, the leaves are light and erect.
6 SOTALIS It is a variety of lettuce with a very nice presentation Enza
for spring and autumn crops. It has medium and Zaden
good disease resistance (Bl: 1-28, Nr: 0).
The head is compact, uniform in light green and
weighing 700-900g. Excellent for marketing and
fresh consumption.

The lettuce was planted on March 6, 2017, the planting scheme being 0.4 x
0.3m = 8plt / m2 of seedlings prepared in the multiplier of the same university.
The behavior of lettuce was followed by the application of gibberellic acid
(AG3) in different doses respectively: gibberellic acid 25mg / l and gibberellic acid
50mg / l applied twice during vegetation.(Table 2).
204
 During the vegetation, phenological observations were made. Biometric
measurements, respectively mean weight of head, weight of the lettuce part, plant
diameter, weight and volume of the roots were recorded at the harvest and production
was recorded.
At the same time, analyzes were made regarding the quality of lettuce, nitrate
content, phosphate nitrate and lettuce potassium. The methods used in the analyzes
were: Griess method, spectrophotometric for nitrates, nitrites and Duval reagent and
spectrophotometric phosphorus dosing, and for flame spectrophotometry. These
methods are according to the standards in our country: ISO 9001.( Metodologia de
analiza a solului si plantei. ICPA, 1987).

RESULTS AND DISCUSSIONS

Table 2
Scheme of experimental variants
No Hibrid Tratment

1 TUAREG Control
2 Gibberellic acid 25mg/l
3 Gibberellic acid 50mg/l
6 ALANIS Control
7 Gibberellic acid 25mg/l
8 Gibberellic acid 50mg/l
9 SHANGORE Control
10 Gibberellic acid 25mg/l
11 Gibberellic acid 50mg/l
12 ANALENA Control
13 Gibberellic acid 25mg/l
14 Gibberellic acid 50mg/l
15 CENTORE Control
16 Gibberellic acid 25mg/l
17 Gibberellic acid 50mg/l
18 SOTALIS Control
19 Gibberellic acid 25mg/l
20 Gibberellic acid 50mg/l

Determination of the average diameters of lettuce showed that the smallest

diameters were obtained at Centore hybrids where the application of gibberellic acid
caused the decrease of this characteristic from 33cm to control at 31cm to 50mg / l
gibberellic acid and 32cm, respectively, at the application of 25mg / l gibberellic
acid.(Figure 1`)
Beneficial results were obtained in several hybrids, namely Shangore, Analena
and Sotalis, at which the lettuce diameters reached 41 cm at the application of 50 mg /
l gibberellic acid.Touareg and Alanis responded differently to the increasing doses of
gibberellic acid, the lettuce diameters increased at 25mg / l and then dropped to the
maximum dose of 50mg / l.
The analysis of lettuce weights in the experimental variants showed differently,
so at a lower dose of 25mg / l we have an increase in weight in all hybrids but then at
50mg / l in a decrease in weights and recorded production respectively.(Figure 2)
The biggest increases in lettuce weight were recorded at Shangore (373g / plt)
and Centore (376g / plt).

205
 Figure 1.The diameter of lettuce in experimental variants

The statistical interpretation of the results (Table 3) shows that the application
of 25 mg / Gibberellic acid resulted in increased production with very significant
differences compared to control, eg Alanis, Shangore and Centore hybrids. When only
50mg / l gibberellic acid was applied, the results were either insignificant (Tuareg,
Shangore) or only significantly statistically ensured.

Figure 2. The medium weight of lettuce plants on variant

At the harvest, lettuce quality, nitrates, phosphates and potassium contents were
also analyzed (Table 4.). Nitrates have been absorbed in varying amounts in plants.
The highest values were obtained when applying 25 mg / l gibberellic acid to Analena
and Sotalis hybrids.( Wang et al., 2002)
If we analyze the results in terms of the Maximum Allowable Concentration at
lettuce cultivated in the greenhouse of 3000 ppm nitrates, the lettuce content of all the
variants under investigation was below this value, so the lettuce is good for
consumption.( Santamaria, 2006). In the case of phosphorus, the absorbed quantities
are small, not exceeding 200 ppm, and the potassium is absorbed in high amounts,
enrolling in the literature values for these elements.

206
 Table 3
Statistical Interpretation of Production Outputs in the 6 Hybrids

No Hybride Tratment Produc- Dife- Percent Signifi-

tion rence % cant
kg/m2 ±
1 TUAREG Control 2.43 - 100.00 -
2 Gibberellic acid 25mg/l 2.82 0.34 116.05 *
3 Gibberellic acid 50mg/l 2.57 0.14 105.76 ns
6 ALANIS Control 2.32 - 100.00 -
7 Gibberellic acid 25mg/l 2.8 0.48 120.69 ***
8 Gibberellic acid 50mg/l 2.72 0.40 117.24 **
9 SHANGORE Control 2.4 - 100.00 -
10 Gibberellic acid 25mg/l 2.99 0.59 124.58 ***
11 Gibberellic acid 50mg/l 2.52 0.12 105.00 ns
12 ANALENA Control 2.3 - 100.00 -
13 Gibberellic acid 25mg/l 2.59 0.29 112.60 *
14 Gibberellic acid 50mg/l 2.84 0.54 123.47 ***
15 CENTORE Control 2.19 - 100.00 -
16 Gibberellic acid 25mg/l 3.01 0.82 137.44 ***
17 Gibberellic acid 50mg/l 2.67 0.48 121.91 ***
18 SOTALIS Control 1.94 - 100.00 -
19 Gibberellic acid 25mg/l 2.28 0.34 117.52 *
20 Gibberellic acid 50mg/l 2.23 0.29 114.94 *
DL 5% = 0.232kg/m2
DL 1%=0.345kg/m2
DL 0.1% = 0.454kg/m2
Table 4.
The lettuce quality characteristics of the experimental variants

No Hybride Tratment N-NO3, P-PO4, K,ppm

ppm ppm
1 TUAREG Control 494 198.12 3940
2 Gibberellic acid 25mg/l 646 152.40 2340
3 Gibberellic acid 50mg/l 760 129.54 5260
6 ALANIS Control 513 251.46 3940
7 Gibberellic acid 25mg/l 646 106.68 3560
8 Gibberellic acid 50mg/l 722 144.78 1880
9 SHANGORE Control 836 182.88 1700
10 Gibberellic acid 25mg/l 418 83.82 2300
11 Gibberellic acid 50mg/l 418 121.92 2100
12 ANALENA Control 760 198.12 2260
13 Gibberellic acid 25mg/l 1120 167.64 2060
14 Gibberellic acid 50mg/l 950 144.78 2300
15 CENTORE Control 646 289.56 2840
16 Gibberellic acid 25mg/l 494 152.4 1640
17 Gibberellic acid 50mg/l 703 121.92 2180
18 SOTALIS Control 513 220.98 2380
19 Gibberellic acid 25mg/l 1254 144.78 2480
20 Gibberellic acid 50mg/l 418 121.92 1500

207
 CONCLUSIONS
1. Determination of average lettuce diameters showed that beneficial results
were obtained in several hybrids, namely Shangore, Analena and Sotalis, at which
lettuce diameters reached 41 cm when applying 50 mg / l gibberellic acid.
2. Touareg and Alanis responded differently to the increasing doses of
gibberellic acid. Lettuce diameters increased at 25mg / l and then dropped to a
maximum dose of 50mg / l.
3. The analysis of the lettuce weights in the experimental variants showed
differently, so at a lower dose of 25mg / l we have an increase in weight in all hybrids
but then to 50mg / l in a decrease in weights and the recorded production respectively.
Lettuce weight gains were recorded at Shangore (373g / plt) and at Centore (376g /
plt).
4. The statistical interpretation of the results shows that the application of 25
mg / Gibberellic acid resulted in increased production with very significant differences
compared to control, eg Alanis, Shangore and Centore hybrides. When only 50mg / l
gibberellic acid was applied, the results were either insignificant (Tuareg, Shangore) or
only significantly statistically ensured.
5. Nitrates were absorbed in varying amounts in plants. The monkey values
were obtained by applying 25 mg / l gibberellic acid to the Analena and Sotalis hybrids.
6. If we analyze the results in terms of the Maximum Allowable Concentration
of lettuce cultivated in greenhouses of 3000ppm nitrates, the lettuce content of all the
variants under investigation is below this value, so the sludge is good for consumption.
7. In the case of phosphorus, the absorbed quantities are small, not exceeding
200 ppm, and the potassium is absorbed in high amounts, enrolling in the values
presented by the literature for these elements.

REFERENCES
Tamme T., Reinik M., Roasto M. Nitrates and Nitrites in Vegetables: Occurrence
and Health Risks. In: Watson R.R., Preedy V.R, editors. Bioactive Foods Promoting
Health: Fruits and Vegetables. Academic Press; Salt Lake City, UT, USA: 2009. pp.
307–321.
Wang Z.H., Zong Z.Q., Li S.X., Chen B.M. Nitrate accumulation in vegetables and
its residual in vegetable fields. Environ. Sci. 2002;23:79–83.
Pavlou G.C., Ehaliotis C.D., Kavvadias V.A. Effect of organic and inorganic
fertilizers applied during successive crop seasons on growth and nitrate accumulation
in lettuce. Sci.Hortic. 2007;111:319–325.
Santamaria P. Nitrate in vegetables: Toxicity, content, intake and EC regulation. J.
Sci. Food Agr. 2006;86:10–17. doi: 10.1002/jsfa.2351.
*** Metodologia de analiza a solului si plantei. ICPA, 1987
*** www.giberelina.ro

208
 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
 Food produce process ing
UNIVERSITY OF CRAIO VA
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

COMPARATIVE STUDY ON POLYPHENOLS CONTENT AND

ANTIOXIDANT ACTIVITY IN SOME SWEET PEPPER CULTIVARS
DURING DEVELOPMENT IN HYDROPONIC CULTURE
1 1 2
Papathanasiou Theodoros , Gougoulias Nikolaos *, Kamvoukou Christina- Anna ,
1 3
Efthimiou Spyridoula , Papachatzis Achilleas
1
Department of Agronomy Technology, Technological Educational Institute of Thessaly, 41110 Lar issa,
Greece
2
School of Pharmacy, Aristotle University of Thessaloniki, Greece
3
School of Agriculture, Aristotle University of Thessaloniki, Greece
*Correspondence author. email: [email protected]

Key words: FRAP activity; Pepper fruits; Stages ripening; Total phenols

ABSTRACT
The changes of the total phenolic content and of the FRAP activity during development and
ripening three sweet peppers cultivars in hydroponic culture they studied. For all cultivars of
sweet peppers studied, from early development stages up to the mature green color, the total
phenols content and antioxidant activity did not show any increase. From mature green color up
to mature red color of fruits, all varieties studied showed a significant increase the levels of the
total phenolics content and FRAP assay values. In particular, total phenolics content for the
Dolmy, Yahoo and Florinis peppers increased by 220%, 195% and 260% respectively. FRAP
assay values for the Dolmy, Yahoo and Florinis peppers were found to be 2-fold, 1.8-fold and
2.9-fold times raised respectively. It is advisable to consume sweet peppers at the mature red
stage for better effects consumer health.

INTRODUCTION
The free radicals are strong oxidisers which damage the biological molecules,
causing the occurrence of many diseases in the human organism (Halliwell 1991).
Studies demonstrate that the oxidative destruction of biomolecules can be protected by
using endogenous and exogenous antioxidants which are contained in fruits,
vegetables etc. (Ou et al. 2002, Pellegrini et al. 2006, Wu et al. 2004).
Phenolic compounds are secondary metabolites which are synthesized by
plants and contained in a high proportion in the plants and the fruits (Gougoulias
2012). Many phenolic compounds contribute to shaping of the taste and color of fruits
and plants, while others are good sources of natural antioxidants and have protective
role against some diseases (Gambacorta et al. 2009, Shahidi 2000). The intake of
polyphenols with food reduces the risk of cardiovascular, carcinogens and other
diseases (Manach et al. 2004, Scalbert and Williamson 2000).
Sweet peppers fruits (Capsicum annuum L.) it is natural reservoir of nutrients
and of natural antioxidants with high antioxidant activity (Gougoulias et al. 2016). The
cultivation systems, fertilization, irrigation and variety, affect the levels of the
antioxidant activity in the peppers fruits (Ayodele et al. 2015, Guinoza et al. 2015).

209
 The aim of this study was the identification of the phenolic content and
antioxidant activity in three sweet peppers cultivars during the development and
ripening, in a hydroponic culture.

MATERIAL AND METHODS

Pepper plants (Capsicum annuum L.) obtained from a commercial nursery of
Larisa, were transplanted and ware cultivated in hydroponic system at the
Technological Education Institute of Larisa, Greece, during February 2016 to July 2016
2
in an area of 200 m . The crop substrate consisted from coconut slabs in double
rows, with a distance of plants for each slab 30 cm. Three cultivars of the sweet
peppers ware cultivated. The pepper fruits and the basic characteristics of each variety
are presented in Table 1.

Table 1
Basic characteristics of pepper fruits

Cultivars Flavor Shape Flesh Color of

thickness harvest
Dolmy F1 Sweet Bell High Green
Yahoo F1 Sweet Elongated High Green
Florinis NS 700 Sweet Elongated High Red

For experimental purposes, drip irrigation technique was used in order to

supply the open hydroponic system. The crop was fertilized through a stably chemical
nutritive solution at the rates of 58.9 ml/min for 3 minutes, repetitively 4 times a day
(Table 2). Nutrients were kept separately according to their chemical properties in
three tanks. The nutritive solution was prepared automatically mixing equal solution
2+
quantity from each tank. The final nutritive solution consisted of Ca = 253.5 meq/L,
K = 380.1 meq/L, Mg = 97.2 meq/L, NH4 = 27.5 meq/L, H = 168 meq/L, Fe2+ =0.9
+ 2+ + +

meq/L, NO3- = 421.9 meq/L, PO43- = 214.9 meq/L and SO42- =290.3 meq/L.

Table 2
Chemical properties of the coco palm substrate and the nutrient solution were used in
experiment

Chemical Property * Coco palm Nutrient Solution Irrigation

(fresh mass) influx water
pH 6.78 ± 0.31 6.45 ± 0.28 7.7
Electrical conductivity, (dS/m) 0.69 ± 0.38 2.23±0.31 0.55
* Coconut pH and electrical conductivity, determined in (1:5) coconut / water
extract. Data represent average means and SE deviation. (n) = 8.

Four pepper fruit for each variety with uniform size, shape and color were
harvested for analysis at each development stage. The development stages and
maturation of pepper fruits are presented in Table 3.
Preparation of the methanol extracts: Ten g of the peppers samples were
two rounds treated by 20 ml of 80% aqueous methanol. Samples were incubated for
24 h in the extractant at stirring; the supernatant material was removed. The pellet was
re-treated with aqueous methanol for 2 h at stirring at ambient temperature. The

210
extract was gathered after centrifugation/filtration and the volume was made up to 50
ml with aqueous methanol and used for further chemical analysis (Kanner et al. 1994).

Table 3
The development stages and ripening of the pepper fruits during cultivation

Stages Cultivars
Dolmy F1 Yahoo F1 Florinis NS 700
Size and color
S1 Small, green Small, green Small, green
S2 Medium, green Medium, green Medium, green
S3 Big, green Big, green Big, green
S4 Maturation, green Maturation, green Maturation, green
S5 30% red 30% reddish-brown 30% red
S6 Full red Full reddish-brown Full red
(S1), 10 days after flowering

Methods of analyses: Total polyphenolic (TP) content was determined with

the Folin-Ciocalteu (F.-C.) reagent according to the method by (Singleton and Rossi
1965) using the microvariant proposed by (Badenschneider et al. 2015) and the
results were expressed as gallic acid equivalent (GAE) in μg/g fresh weight.
The antioxidant activity of ferric reducing antioxidant power (FRAP) of the
methanol extracts was determined on the basis of the method by (Benzie and Strain
1999) and was expressed as ascorbic acid equivalent (AAE) in µmol/g fresh weight
Statistical analysis: Data were analyzed using the MINITAB (Ryan et al.
2005) statistical package. The experiment had four replications. Analysis of variance
was used to assess treatment effects. Mean separation was made using Tukey‟s test
when significant differences (P=0.05) between treatments were found.

RESULTS AND DISCUSSIONS

Among the sweet peppers cultivars studied the six stages development, in
the bell Dolmy pepper the total phenols content ranged from 432.2 to 1380.5 μg (GAE)
/ g FW, in the elongated Yahoo pepper the total phenols content ranged from 377.5
to 1256.9 μg (GAE) / g FW, and in the elongated Florinis pepper the total phenols
content ranged from 540.8 to 2283.8 μg (GAE) / g FW (Table 4).
The Dolmy and Yahoo cultivars from the start of development of fruits (first
stage) up to the mature green color (fourth stage) they showed reduction of the total
phenols content. The total phenols content in the Florinis peppers did not show
statistically significant differences from the start of development of the fruits up to the
mature green color. Subsequently, from mature green color up to mature red color of
fruits, all varieties studied showed a significant increase the levels of the total phenolics
content. The composition of total phenols in the Dolmy, Yahoo and Florinis peppers
increased by 220%, 195% and 260% respectively. To the same conclusions arrived
some researchers which studied the change in the content total phenolic during
ripening for some sweet pepper cultivars of India (Deepa et al. 2007).
Finally, during the mature green color of the sweet peppers or of the mature
red color, the Florinis variety showed a higher total phenols content compared to
Dolmy and Yahoo varieties in the respective stages (P < 0.05). Also, our results at the
stage of full maturation (mature red color) have higher values than those reported by
211
other authors for some hot peppers cultivars of Greece (Gougoulias et al. 2017). These
differences, in the variability of total phenols, likely due to the different cultivars, in soil
and climatic conditions of each region.
Table 4
Changes of the total phenolics content during development and ripening of sweet
pepper cultivars

Development stages and ripening TP-μg (GAE)/g FW

Dolmy F1 Yahoo F1 Florinis NS 700
S1 (small size) 615.9b 512.1c 607.1bc
S2 (medium size) 458.1d 433.8d 665.6b
S3 (big size) 509.4c 377.5e 540.8c
S4 (maturation green) 432.2d 426.9d 611.6bc
S5 (30% red color) 621.6b 578.8b 2283.8a
S6 (full color) 1380.5a 1256.9a 2197.2a
For each different variety of pepper fruits, columns of table with the same letter do not
differ significantly according to the Tukey‟s test (P=0.05).

The results of the FRAP antioxidant assay of three sweet pepper cultivars
studied, reveal that the antioxidant activity FRAP from the start of the development of
fruits (first stage) up to the mature green color (fourth stage), showed no statistically
significant difference for all cultivars studied (Table 5). Significant increase of the
FRAP activity in all sweet peppers cultivars reveal from the mature green color until
the mature red color. FRAP assay values for the Dolmy, Yahoo and Florinis peppers
were found to be 2-fold, 1.8-fold and 2.9-fold times raised respectively. To the same
conclusions arrived some researchers which studied the change in the antioxidant
activity FRAP during ripening for some sweet pepper cultivars of iran
(Ghasemnezhad et al. 2011). Also, the ferric reducing antioxidant power (FRAP) of
sweet peppers reveal that the Florinis cultivar in the mature red color, exert the
highest activity (as ascorbic acid equivalent) with 23.8 μmol (AAE)/g FW, compared to
the other sweet peppers cultivars (P < 0.05).
Table 5
Changes of the antioxidant activity FRAP during development and ripening of sweet
pepper cultivars

Development stages and FRAP activity -μmol (AAE)/g FW

ripening
Dolmy F1 Yahoo F1 Florinis NS 700
S1 (small size) 8.2bc 7.5b 7.9b
S2 (medium size) 7.2c 6.7c 8.8b
S3 (big size) 7.6c 6.5c 7.7b
S4 (maturation green) 7.1c 7.4b 8.3b
S5 (30% red color) 8.8b 7.9b 23.2a
S6 (full color) 14.3a 13.1a 23.8a
For each different variety of pepper fruits, columns of table with the same letter do not
differ significantly according to the Tukey‟s test (P=0.05).

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 CONCLUSIONS
The results of the hydroponic cultivation showed that in the Dolmy, Yahoo and
Florinis sweet peppers, from early development stages up to the mature green color,
the total phenols content and antioxidant activity did not show any increase.
For all cultivars of sweet peppers studied, from the mature green color to the
mature red color, the composition of total phenols and FRAP assay values they
increased significantly. Finally, it is advisable to consume sweet peppers at the mature
red stage for better effects consumer health.

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 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
 Food produce process ing
UNIVERSITY OF CRAIO VA
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

INFLUENCE OF GROWTH REGULATORS ON PRODUCTIVITY OF

PLANTATION AND QUALITY OF APPLE FRUITS

Pesteanu Ananie1
1
Faculty of Horticulture, State Agrarian University of Moldova
Mircesti str. 48, Chisinau, Republic of Moldova; [email protected]

Key words: Apple, economic efficiency, profitability, relative efficiency.

ABSTRACT
The experimental plot is placed in the orchard “Codru-ST” Ltd. founded in 2012 year.
The study subject of the experience was Idared apple variety grafted on M 9 rootstock. The
distance of plantation is 3.5x 0.8 m. The research was conducted during the period of 2015 year.
The tested agents was Humifield, WG, 0.2/0.2/0.2/0.2/0.2/0.2 kg/ha; Nertus PlantaPeg,
0.6/0.6/2.0 l/ha; Nertus PlantaPeg, 1.0/1.0/2.5 l/ha, which was sprayed in different period. During
the research, it was studied the influence on the growth regulators on productivity of plantation
and quality of apple fruits. It was established that the The Nertus PlantaPeg growth regulator can
be included in the technology system to improve apple growth and fructification processes and
reduce the etiquette release rate of fruit, applied 3 times by foliar spraying. The first treatment to
be carried out in the "pink button" phase at a dose of 0.6 - 1.0 l/ha, the second - in the binding
phase - the intensive growth of fruits (fruit diameter 20-25 mm) at a dose of 0.6 - 1.0 l/ha, and
the third - 1-2 days before harvesting in the dose 2.0 - 2.5 l/ha.

INTRODUCTION
The action of the production factors is generally limited after achieving a
certain level of production (Balan et al., 2001). Thus, mineral fertilizers can be
administered up to a certain limit, over which there are regression phenomena of
increased production, water can be administered up to the physiological limit specific to
apple culture. That is why new and new means discovered by man are needed for the
continuous growth of productions (Babuc, 2012; Cimpoieş, 2012).
The discovery of growth-enhancing substances (phytohormones, growth
regulators, etc.) has made it an effective tool for producers to manage and control
growth, plant development and apple production (Babuc et al., 2013; Suman et al.,
2017).
In addition to regulating growth and development processes, growth regulators
also influence other biochemical processes and biological phenomena such as
flowering and fertilization of flowers, fruit binding, yield of photosynthesis, differentiation
of fruit buds, resistance to fall of leaves and fruit (Ghena et al., 1999).
Polyethylene glycol, fulvic acide and salts of humic acids are those compounds
that increase the resistance of trees during vegetation to stress factors such as
drought, high temperatures, insufficiency of mineral elements, disease of various
diseases and pests. The products in question allow their action to have a positive
effect for a longer period after the treatments (Burzo et al., 1999).
Fulvic acids and salts of humic acids improve the activity of the root system in
215
the soil, transforming the minerals that are harder to reach for the trees, in an easily
added form, which through the absorbent roots are supplied to the air side (Burzo et
al., 1999; Neamţu et al., 1991).

MATERIALS AND METHODS

The research was carried out during the year 2015 in the superintensive apple
orchard founded in the autumn of 2012 at the enterprise "Codru ST", with "knip boom"
crown trees.
The object of study of the experience was Idared apples grafted on the weak M9
rootstock. The crown was driven after the improved thinning system. Planting distance
was 3.5 x 0.8 m.
To determine the influence of growth regulators on fructification and not to
imbalance the physiological processes inside the plant and to obtain constant and
quality productions, the following variants were tested (tab. 1).

Table 1
Scheme of experiments to determine the biological efficiency of the growth regulator in
apple trees

Variants Application method

Control - without treatment -

Spraying 6 times. The first treatment - until

Humifield, WG, 0.2/0.2/0.2/0.2/0.2/0.2
flowering, II - after flowering, the next 4 with 10-
kg/ha
14 days between treatments.

Nertus PlantaPeg, 0.6/0.6/2.0 l/ha Spraying 3 times. The first treatment - in the
pink "button" phase, the second - in the binding
phase - the intensive growth of the fruit (fruit
diameter 20-25 mm), and the third - 1-2 days
Nertus PlantaPeg, 1.0/1.0/2.5 l/ha before the harvest.

Trees treatment was done with the portable sprinkler in the hours without wind,
in the morning.The amount of solution per tree when treated with growth regulators on
apple trees was 0.3 liters, based on the number of trees per unit area and the
recommended water quantity of 1000 l/ha.
The amount of fruits, the average weight of a fruit, the production of a tree and
a surface unit, their redistribution by diameter were established during the harvest
period. The results were reported in the control variant and the standard version. The
apple firmness evaluation was performed using the FT 327 penetrometer, which fixes
the opposing pulp resistance to the penetration of a 1 cm2 plunger. The degree of
emission of ethylene was determined using the ICA 56 ethylene analyzer.
The main results obtained were statistically processed by the dispersion
analysis method after Б. Доспехов.

RESULTS AND DISCUSSIONS

Fruit production is one of the main indexes that are taken into account in
determining the efficiency of one or another technological element applied in the
superintensive culture system.
216
 The investigations show that the number of fruits in the crown of the trees
included in the research differs on the variants in the study (tab. 2). If, in the control
variant, the number of fruit was 52 pcs/tree, then, in the variant treated with the growth
regulator Humifield WG in dose of 0.2 kg/ha was 54 pcs/tree. When treating variants
with the Nertus PlantaPeg growth regulator, the studied index consisted of 55 - 56
pieces/tree.
The difference between the control variant and the variant treated with
Humifield WG in dose of 0.2 kg/ha was 2 pcs/tree, but between the third and fourth
variants, which was treated with the Nertus PlantaPeg growth regulator, respectively 3
and 4 pieces/tree.
This increase in the number of fruits in the tree crown was due to improved
metabolism of growth hormones, positive influence on inflorescence fertility, fertilization
and fruit binding process following treatment with Humifield WG and Nertus PlantaPeg
growth regulators during the pink button period and after flowering.
The average weight of a fruit on variants in the study underwent changes
compared to the control variant. The lowest average fruit weight was recorded in the
control variant, where the study index was 173.9 g. When using the Humifield WG
growth regulator in dose of 0.2 kg/ha, the average weight of a fruit constituted 176.3 g,
or an increase of 2.4 g compared to the control variant, without treatment.

Table 2
The influence of the growth regulator on the average quantity, average weight and fruit
production of Idared apple trees
Fruit production In %,
Fruit
Average compared
Variants quantity,
weight, g kg/tree t/ha to the
buc/tree
witness
Control 52 173,9 9,04 32,28 100,0

Humifield, WG,
54 176,3 9,52 34,00 105,5
0,2 kg/ha

Nertus PlantaPeg,
55 179,5 9,87 35,24 109,1
0,6 l/ha

Nertus PlantaPeg,
56 178,5 9,99 35,69 110,5
1,0 l/ha

DL 5% 2,4 7,3 0,45 1,63 -

The use of the Nertus PlantaPeg growth regulator, which besides fulvic acids
and salts of humic acids in its composition also has polyethylene glycols (PEG 400 and
PEG 1500) has positively influenced the average fruit weight. In the third variant,
treated with the Nertus PlantaPeg growth regulator, the average weight of a fruit was
179.5 g, or an increase of 5.6 g compared to the control variant and 3.2 g with the
standard Humifield WG in the dose 0.2 kg/ha. The increase in the Nertus PlantaPeg
growth regulator dose, the fourth variant, lowered the average weight of a fruit to 178.5
g or 0.6% compared to the third variant, but it recorded higher values compared to
control variant by 2.6%.
This slight difference in the average weight in variants 3 and 4 is due to
approximately the same number of fruits in the apple tree crown and maintenance of
217
the physiologically active balance following treatment with the Nertus PlantaPeg
growth regulator. The results outlined above are also confirmed by statistical
processing.
The fruit production per tree and on a surface unit correlates directly with the
number of fruits in the tree crown and their average weight. The smallest fruit
production was recorded in the control variant, constituting 9.04 kg/tree or 32.28 t/ha.
When treatment with the Humifield WG growth regulator was made, the fruit
production was 9.52 kg / tree or 34.00 t/ha, or a 5.5% increase compared to the control
variant without treatment.
The largest fruit production was recorded in the variants treated with the
Nertus PlantaPeg growth regulator. If in the third variant the fruit production constituted
9.87 kg/tree, then in variant four the studied index increased to 9.99 kg/tree, or 1.2%.
The recorded fruit production at a surface unit on the above mentioned variants
constituted 35.24 and 35,69 t/ha respectively. Treatments with Nertus PlantaPeg
growth regulator increased fruit production by 9.1-10.5% compared to control variant
and 3.6-5.0% versus Humifield, WG.
The insignificant difference between the third and fourth variants treated with
the Nertus PlantaPeg growth regulator is also provided by statistical data.
The results show that the Nertus PlantaPeg growth regulator used to prevent
water stress, plant temperature oscillations and maintaining the physiological balance
between growth and fructification in both doses has positively influenced the number of
fruits in the tree crown, the average weight of a fruit and plant productivity.
It is very important that apples are harvested at the optimum time. The data of
the investigations carried out (tab. 3) shows that the firmness of the apple pulp on the
variants studied at the time of the harvest constituted 7.2-7.6 kg/cm2. The smallest
firmness of the pulp was recorded in the control variant without treatment - 7.2 kg /
cm2.
When treating growth regulators, the pulp firmness increased. In the variant
treated with Humifield, WG growth regulator, the firmness of the pulp increased and
constituted 7.4 kg/cm 2, or 0.2 kg/cm 2 higher compared to the control variant. In the
variants treated with Nertus PlantaPeg, it was recorded, approximately the same
values of pulp firmness, 7.6 kg/cm 2, or an increase of 0.4 kg/cm2 compared to the
control variant and 0.2 kg/cm 2 versus the Humifield, WG variant. The increase in the
dose did not affect the firmness of the pulp.
The dry substance content of the Idared variety, on variants in the study, is
15.4 - 15.8%. The smallest value of dry substance weight was recorded in the control
variant - 15.4%.
The treatments with growth regulators increased the index in the study. In the
variant treated with Humifield, WG, the amount of dried substance in the fruit
constituted 15.7%, or an increase of 0.3% compared to the control variant.
When using the Nertus PlantaPeg growth regulator at 0.6 / 0.6 / 2.0 l/ha and
1.0 / 1.0 / 2.5 l/ha, the amount of dried substance in the fruits was 15, 8%. Treatments
on both variants with the Nertus PlantaPeg growth regulator increase 0.4% of the dry
substance content compared to the control variant and 0.1% versus the standard
variant. Therefore, the standard treatments with the growth regulator have influenced
the increase in the amount of dry substance in the fruit.
The content of soluble dry substance is a sort of variety, after which the
optimum harvesting time can be determined. The investigations show that the quantity
of soluble dry substances in the Idared variety on the variants in the study constituted
13.6 - 14.0%.

218
 Table 2
Influence of the growth regulator on the firmness, biochemical indices and the amount of
ethylene emanated from Idared apples

The
Dry Soluble dry amount of
Firmeness, Titratable
Variants substance substance, ethylene
kg/cm2 acidity,%
,% % emitted,
µl/kg/h
Control 7,2 15,4 13,6 0,50 0,013

Humifild, WG, 0,2

7,4 15,7 14,0 0,47 0,012
kg/ha

Nertus Planta Peg,

7,6 15,8 13,9 0,46 0,003
0,6 l/ha

Nertus Planta Peg,

7,6 15,8 14,0 0,46 0,004
1,0 l/ha

The smallest value of the weight of the soluble dry substances was recorded in
the control variant without treatment - 13.6%. In the case of treatment with growth
regulators, we record an increase in the study index to 13.9 - 14.0%. That is, when
treated with the Humifield, WG standard growth regulator, the amount of fruit-soluble
dried substance increases by 0.4% compared to the control variant.
Treatments with the Nertus PlantaPeg growth regulator on both variants
increases by 0.3 - 0.4% compared to the control variant, and compared to the standard
variant the same values were recorded. Therefore, treatments with growth regulators in
the study have positively influenced the increase in the amount of soluble dry
substance.
The share of titratable acids in fruits is in direct dependence on the amount of
soluble dry substance. Concomitantly, with the decrease of the quantity of dry
substances soluble in fruits the share of titratable acids increases, registering the
highest value in the control variant - 0.50%.
When, the treatment was made with the Humifield, WG growth regulator, the
amount of titratable acid in the fruit constituted 0.47%, and with Nertus PlantaPeg
decreased, constituting 0.46%.
Ethylene is considered as the main fruit maturing hormone. This substance is
synthesized in all plant organs and in larger quantities in apples.
The amount of ethylene emitted by fruit immediately after harvest differs on the
variants in the study. The largest quantity of ethylene emitted was recorded in the
control variant without treatment - 0.013 μl/kg/h.
In the variant treated with the Humifield, WG growth regulator, the amount of
ethylene emitted by the fruits is 0.012 μl/kg/hr.
When using the tested growth regulator Nertus PlantaPeg, which has the
ability to form a protective film on the fruit, the amount of ethylene emitted from fruits
has diminished on the variants in the study up to 0.003 - 0.004 μl/kg/hr.
This demonstrates that treatment with the Nertus PlantaPeg growth regulator
inhibited the amount of ethylene emitted by fruit by 0.009-0.010 μl/kg/hr compared to
the control variant. Increasing the dose of Nertus PlantaPeg has insignificantly
influenced the indicator in the study.
219
 CONCLUSION
The Nertus PlantaPeg growth regulator can be included in the technology
system to improve apple growth and fructification processes and reduce the etiquette
release rate of fruit, applied 3 times by foliar spraying. The first treatment to be carried
out in the "pink button" phase at a dose of 0.6 - 1.0 l/ha, II - in the binding phase - the
intensive growth of fruits (fruit diameter 20-25 mm) at a dose of 0.6 - 1.0 l/ha, and III -
1-2 days before harvesting in the dose 2.0 - 2.5 l/ha.

REFERENCES
Babuc V., 2012, Pomicultura. Edit. Tipografia Centrală, Chişinău, 662 pag.
Babuc V., Peşteanu A, Gudumac E., Cumpanici A., 2013, Producerea merelor.
Edit. Bonss Office, Chişinău, p.158-169.
Balan V., Cimpoieş Gh., Barbăroşie M., 2001, Pomicultura. Edit. Museum,
Chişinău, 453 pag.
Burzo I., Toma S., Olteanu I., Dejeu L., Delian Elena., Hoza D., 1999,
Piziologia plantelor de cultură. Fiziologia pomilor fructiferio şi a viţei de vie. Edit. Ştiinţa,
Chişinău, (3), p. 9-234.
Cimpoieş Gh., 2012, Cultura mărului. Edit. Bons Offices, Chişinău, 380 pag.
Ghena N., Branişte N. Stănică Fl., 2004, Pomicultura generală. Edit.
MatrixRom, Bucureşti, p. 391-422.
Neamţu G., Irimie Fl.,1991, Fitoregulatori de creştere. Edit. Ceris, Bucureşti,
336 pag.
Suman M., Sangma P., Meghawal D., Sahu O., 2017, Effect of plant growth
regulators on fruit crops. Journal of Pharmacognosy and Phytochemistry, 6(2): 331-
337.

220
 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
 Food produce process ing
UNIVERSITY OF CRAIO VA
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

POSSIBILITIES OF OPTIMIZATION OF THE COMPLEX FERTILIZATION IN

CORNICHON CUCUMBERS CROP

Petcuci Andrei-Cătălin1, Atanasiu Nicolae1, Neață Gabriela1, Câmpeanu Gheorghe1,

1 2
Cornea Călina Petruța , Petcuci Alexandra
1
University of Agronomical Sciences and Veterinary Medicine Bucharest, Horticulture Department,
Bucharest, Romania,
2
Dow AgroSciences
Corespondence author: E-mail:[email protected]

Key words: cornichon cucumbers, fertigation, complex fertilization

ABSTRACT
Cucumber is a vegetable appreciated and wanted very much not only by consumers
but also by the farmers. In the evolution of the cultivated area, cucumbers have gained the
extension of the protected crop even at higher altitude, using the conditions of the appearance
and extension of plastic-houses or cold or warm glass-houses. These culture systems are
appreciated by the farmers due to short length of the crop cycles that occupy these constructions
not more than 2.5-4 months in case of cornichon cucumbers. For obtaining the maximum of
production and quality of the cornichon cucumbers crop, very important is to establish a
fertirigation of a complex fertilization program that is efficient and optimum depending on the
crop characteristics.

INTRODUCTION
Cucumbers are known and cultivated in southern and south-east Asia and
northern Africa for more than 4000 years. Nowadays, due to the technological
evolution, cucumbers are grown on large geographic areas between the Equator and
the large latitudes of the Scandinavian countries (for example in Finland).
Resistance and / or tolerance to diseases is a technological asset of
cucumbers that can reduce the number of phytosanitary treatments, while lowering the
costs of these works, resulting in a production with fewer inputs and a higher
profit.(Atanasiu, 1999, Popescu et al., 2002)
Elasticity of production is also remarkable for small fruit cucumbers: fruits of
this species can be harvested when they reach either very small size (3-5 cm long) or
large size (over 10-14 cm). From a crop of cornichon cucumbers, due to this
technological trait, fruits intended for industrialization or fruit intended for fresh
consumption can be harvested.

MATERIAL AND METHODS

The following variants were studied in order to develop the previously
presented and argued experience.
Experience has been assembled on two sites:
- On the ground, in a cold plastic house, in Brezoaele, Dâmboviţa County

221
 - On peat used as rootstock in an unheated greenhouse from the technical
equipment of the sector of greenhouses District 4, ADP Bucharest.
The experience was located in a traditional tunnel-type plastic house, with a
base width of 5.40 m and in a cold greenhouse. The plastic house, located in the
Brezoaele area (Dâmboviţa County), is located on brown-red soil, with a moderate
degree of favorability for the cultivation of cornichon cucumbers. In the cold
greenhouse, District 4, Bucharest, the peat rooting substrate was used.
At the same time, analyzes were made regarding the quality of lettuce, nitrate
content, phosphate nitrate and lettuce potassium. The methods used in the analyzes
were: Griess method, spectrophotometric for nitrates, nitrites and Duval reagent and
spectrophotometric phosphorus dosing, and for flame spectrophotometry. These
methods are according to the standards in our country: ISO 9001.( Metodologia de
analiza a solului si plantei, ICPA, 1987).
Table 1

Experimental variants. Influence of the assortment and the culture substrate on

the production of cornichon cucumbers in cold plastic houses in autumn culture

Var. no. Culture substrate Variety

V1(mt) Kybria F1
V2 On soil, in plastic house Lenara F1
V3 Mirabelle F1
V4(mt2) Kybria F1
V5 On peat, in cold glasshouse Lenara F1
V6 Mirabelle F1

Basic fertilization was carried out in the autumn by the application of 3 kg of

well fermented manure to square meters of land.
Cornichon cucumber culture was established at 80/40 cm distances, providing
a density of 3.15 plants/m 2 in the case of the plastic house, and at distances of 160
(between peat bags) / 40 cm, with 2 plants per the peat bag, which ensures the same
density. This density has been established since the very good light conditions in the
first part of the crop cycle favor both vegetative growth and fructification of cucumbers.
Phaseal fertilization during vegetation was applied to the soil, along with
watering, using recommended doses of the Universol, richer in nitrogen before fruiting
and richer in potassium during the fruiting period.
The plastic house was not equipped with water spray equipment to increase
atmospheric humidity and maintain it inside the building. The consequence of this
situation was unfavorable for culture, influencing negatively the level of production
(Horgos, 1999).

RESULTS AND DISCUSSIONS

Additional fertilization has a special contribution. The first one is done 10-15
days after the rising, and the second one 15-20 days after the first.
The best soils for cucumber culture are fertile, loose, permeable soils. For
early production, the most obvious are light soils. Heavy, clay and soils are unsuitable.
Also avoid soils too wet or too dry and those that make the crust.

222
 Cucumber culture soil should have a good structure; high content of organic
matter, with groundwater at 80-90 cm. Ground water should not be over 70 cm. The
soil reaction must be slightly acidic to neutral (pH 6.5-7).
Soil fertilization for cucumber cultivation is indispensable. The type and
amount of fertilizer depends on the type of soil and its natural fertility. Cucumbers in
general need a lot of organic matter. That is why large crops are obtained at a massive
fertilization (40-60 tones / ha) with well-decomposed waste. Poorly decomposed
garbage cannot be used by cucumbers unless it was given to the precursor crop. in
this case, the fertilization is supplemented with 250-300 kg of superphosphate, 100-
150 kg of ammonium nitrate and 150-200 kg of potassium salt. The need for nutrients
is high in cucumbers and cannot be covered only by mineral fertilization.
The cucumber requirements for nutrients are as follows:
• Medium requirement for phosphorus;

• High demand for potassium and nitrogen.

Figure 1 – Traditional plastic house, Figure 2 – Cold glasshouse,
Brezoaele, Dâmboviţa county District 4, Bucharest

223
 Cucumber culture cannot be practiced on the same field for several
consecutive years or after other cucurbits, as soil fatigue occurs very rapidly and
disease and pests are greatly multiplied. It is recommended not to return to the same
place with a cucumber culture until after at least 4 years. It is recommended a place for
cucumbers where last year crops have been planted that improve the soil structure,
then those that leave free-weeds soils like onions, potatoes, tomatoes, late beans, then
spinach and early salad.
To achieve the described experience, the following complex fertilizers were
used:
Table 2

Some Chemical and Technological Particularities of Complex Fertilizers

Universol Albastru and Universol Violet, Brezoaele, Dâmboviţa County, 2016
Composition %

Macroelements Macroelements
No
Universol albastru Universol violet
1. Total N – 18, from which 10, from which
N-NO3 – 10.0 7.2
N-NH4 – 7.7 3.0
2 Urea (when necessary) – 0.5 0.5 (when necessary)
3 P2O5 – 11 10
4 K2O – 18 30
5 MgO – 2.5 3.3
6 SO4 – 8.3 15.5
7 CaO – 0.0 0.0
Microelements Microelements
Universol Albastru Universol violet
8 Bor – 0.01 0.01
9 Cu – 0.01 0.01
10 Fe – 0.10 0.10
11 Mg – 0.04 0.04
12 Mo – 0.001 0.001
13 Zn – 0.01 0.01

E.C. la 21oC – 1.3 1.5

o
Maximum solubility at 25 C – 350 300

Recommended dose rates: 10-15 g/m2/week

224
 Table 3
Nutrient content in differentiated nutrient solution on phenophases
A - cornichon cucumber in cold greenhouses on peat substrate, autumn culture,
Phenophase I - emergence - first harvest,
complex fertilizer – Universol albastru
Total MgSO
Concen- CaNO3 Concen-
Universol KNO3 4
No Nutrient tration 0.50 tration
albastru, 0.35 g/L 0.25
mg/L g/L mg/L
1 g/L g/L
1 N (total) 180-225 180 0.35 80 - 305
2 K 225-300 180 160 - - 340
3 Ca 145-185 - - 137 - 137
4 Mg 38-42 21 - - 40 61
5 P 40-50 110 - - - 110
6 SO4 40-60 - - - 80 80
7 Fe 0.06 0.06 - - - 0.06
8 B 0.01 0.01 - - - 0.01
9 Cu 0.01 0.01 - - - 0.01
10 Mn 0.04 0.04 - - - 0.04
11 Mo 0.001 0.001 - - - 0.001
12 Zn 0.001 0.001 - - - 0.001

Table 4
Nutrient content in nutrient solution differentiated on phenophases
- B - cornichon cucumber in cold greenhouses on peat substrate, autumn
culture, Phenophase II - from first to last harvest,
complex fertilizer – Universol violet

Total Conce
Concen- KNO3 CaNO3
Universol MgSO4 n-
No. Nutrients tration 0.35 0.50
violet, 0.25 g/L tration
mg/L g/L g/L
1 g/L mg/L
1 N (total) 180-225 90 45 50 - 185
2 K 225-300 270 160 - - 430
3 Ca 145-185 - - 137 - 137
4 Mg 38-42 33 - - 40 73
5 P 40-50 90 - - - 90
6 SO4 40-60 - - - 80 80
7 Fe 0.06 0.06 - - - 0.06
8 B 0.01 0.01 - - - 0.01
9 Cu 0.01 0.01 - - - 0.01
10 Mn 0.04 0.04 - - - 0.04
11 Mo 0.001 0.001 - - - 0.001
12 Zn 0.001 0.001 - - - 0.001

225
 The nutrient solution plant has three pools:
Basin A - alkaline reaction
- Calcium nitrate - Ca - Ca (NO3) 2 - additional + urea + ammonium nitrate -
max 15%
- Potassium nitrate - KNO3
- Fe - chelated - DTPA
Basin B - acidic reaction
- Potassium nitrate - KNO3
- Potassium sulphate - K2SO4
- Magnesium sulphate - MgSO4
- Magnesium nitrate - Mg (NO3) 2
- Monopotassium phosphate - KH2PO4
- Ammonium nitrate - NH4NO3
- Phosphoric acid - H3PO4 - or nitric acid
- Microelements - manganese sulphate
- copper sulphate
-boron
-molybdate sodium
Basin C - nitric acid
In the Brezoaele experience, fertirigation and Universol Galben were used in
the first vegetation phases at up to 10 g/m2/ week (3 g / plant) - this replaces classical
fertilization with phosphorus chemical fertilizers.
Harvesting must be done when cucumbers are young. For pickled cucumbers
are indicated fruits of 8-12 cm length. Harvesting is repeated at 2-3 days, because the
fruit ages very quickly and loses its quality. You do not have to rotate or twist gobs
because the plant suffers. If they are left unharvested and aged, the plants no longer
bind fruit.
From 1000 m2 of culture we can get 1000-2000 kg of early cucumbers, 2000-
3000 kg of summer cucumbers and 1500-2000 kg of autumn cucumber.
Comparing the results obtained with the three hybrids cultivated on the soil, we
can see that the best results were obtained at Kybria with a production of 12.9 kg / m 2
The production at Lenara and Mirabelle came close to Kybria's production on
the ground.
Comparing the results obtained with peat culture all Kybria has the highest
production of 17.40kg / m 2.
Comparing the experimental variants cultivated on the soil with the peat
cultivars, it can be seen that all peat variants yielded distinct significant results at
Mirabelle and very significant in Kybria and Lenara.

CONCLUSIONS
1. Comparing the results obtained with the three hybrids cultivated on the soil,
we can see that the best results were obtained at Kybria with a production of 12.9 kg /
2
m
2. The production at Lenara and Mirabelle came close to Kybria's production
on the ground.
Comparing the results obtained with peat culture all Kybria has the highest
2
production of 17.40kg / m .
3.Comparing the experimental variants cultivated on the soil with the peat
cultivars, it can be seen that all peat variants yielded distinct significant results at
Mirabelle and very significant in Kybria and Lenara.

226
227
 REFERENCES
Atanasiu N.-1999.Contributii la imbunatatirea tehnologiei de cultura a castravetilor
tip cornichon in camp in sistem industrial-Teza de doctorat-USAMV Iasi.
Popescu V.,Atanasiu N.-2002.Legumicultura-vol III,Ed.Ceres,Bucuresti.
*** Metodologia de analiza a solului si plantei. ICPA, 1987

228
 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
 Food produce process ing
UNIVERSITY OF CRAIO VA
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

INFLUENCE OF SEVERAL TECHNOLOGICAL LINKS APPLIED IN THE

ECOLOGICAL SYSTEM ON THE QUALITY OF GRAPES PRODUCTION

Pîrcălabu Liliana1*, Tudor Georgeta, Ion Marian

1*
Research Institute for Viticulture and Enology Valea Călugarească
* Correspondence author. E-mail: [email protected]

Key words: soil maintenance systems, black furrow, total mulching

ABSTRACT
The researches were carried out in two vineyards grown with the Feteasca neagra and
Cabernet Sauvignon varieties, with two experimental factors: the soil maintenance system (black
furrow, total mulching with straws, partial mulching with marc compost, permanent herbage with
grasses spontaneous flora) and the system for the control of pests and diseases (ecologically
and conventional). In the Feteasca neagra variety, the highest yields of 3.83 kg/vine (ecological
control system) were obtained in the case of soil maintenance by mulching with marc compost
compared with 4.23 kg/vine (conventional control system). In terms of total mulching with straws
the production was 3.76 kg/vine (ecological control system), compared to 4.08 kg/vine
(conventional control system). The black furrow system has produced a yield of 3.71 kg/vine
(organic control system) compared to 3.89 kg/vine (conventional control system)..

INTRODUCTION
In accordance with the concept of organic viticulture allocation technological
inputs should be performed at a level that will produce reasonable yields, while
ensuring adequate protection of the ecosystem vineyard.
The viticultural ecosystem has evolved over time to an anthropic ecosystem
where the anthropic factor has gained a more and more obvious regulatory function. In
these conditions, the functioning of the viticultural ecosystem and implicitly its
productive capacity can be negatively affected by the anthropic factors, represented by
the applied crop technologies (Dejeu and Matei, 1996, Heidi Resenthal Duminy,
2004)).
Different soil maintenance systems (mechanically, manually, biologically) and
to control vines and pests (ecologically, conventionally), each with their advantages
and disadvantages, generates in time differentiated effects on the soil, the plant and
the environment, in the production of grapes obtained and in their quality and
sanogenity.

MATERIAL AND METHODS

The researches were carried out in two vineyards grown with the Feteasca
neagra and Cabernet Sauvignon varieties, with planting distances of 2 x 1 m and the
bilateral Cordon form. The cutting system practiced is mixed in two-three eyes taps
and a cutting load of 10-12 eyes per vine, with 12-15 eyes/square meter loadings.

229
 The soil in the experimental polygon is eumezobazic brown, with a sandy loam
texture, pH weak acid (6.1), well supplied with humus (2.9%) and useful mineral
elements (N, P, K).
The device included two experimental factors: the soil maintenance system
(black furrow, total mulching with straws, partial mulching with marc compost,
permanent herbage with grasses spontaneous flora) and the system for the control of
pests and diseases ecologically (E) and conventionally (C).

RESULTS AND DISCUSSIONS

The experimental results obtained showed that under the ecoclimatic
conditions specific to 2016 year, characterized by a high heliothermic regime of rich
water resources, the experimental technological variants had a different impact on the
quality of the grape production.
Compared to normal (1985-2015), the average monthly air temperature varied
between + 3.0°C (April), + 0.6°C (Jun), and -1.9°C (May).
The vegetation period (April) started with average temperatures higher than
normal (14.7°C to 11.7°C) and a high water regime (65 l/square meter to 44.8 l/square
meter) multiannual (Table 1).
Table 1
The thermal regime during April-October

Monthly average Maximum Minimum

Month temperature absolute absolute
temperature temperature
Normal 2016
April 11.7 14.7 29.3 3.6
May 17.5 15.6 29.2 6.1
June 21.5 22.1 33.8 10.5
July 23.6 23.7 33.4 14.4
August 23.3 22.7 35.0 12.2
September 18.1 19.0 32.1 6.5
October 12.2 12.1 25.4 5.4

In 2016, the air hygroscopicity was higher by 6.6% in May, 2.2% in June, 5% in
October and lower hygroscopicity by 0.7% in April, 1.1% in August, 5.6% in September
and the amount of monthly rainfall recorded to the normal (1985-2015) positive
differences 20.2 mm (April) 14.9 mm (May), 15.5 mm (August) and 11.6 mm
(September) and negative difference of -5.6 mm (July), (Table 2).

Table 2
The pluviometric regime during April-October

Rainfalls Number of days

Month with rainfall
above 10 mm
Normal 2016
April 44.8 65.0 2
May 67.3 82.2 2
June 81.5 81.8 5
July 75.8 70.2 2
230
 August 62.7 78.2 4
September 54.4 66.0 3
October 46.2 120.0 4

Thus, with regard to the control systems under study, the use of conventional
pest and disease pest control has ensured the highest yields. The average weight of a
grape was varied with oscillations between 93.65 g (conventional control system-C)
and 94.43 g (ecological control system-E) the Feteasca neagra and Cabernet
Sauvignon range between 93.57 g (conventional control system-C) and 84.75 g
(ecological control system-E), (Table 3).

Table 3
Production of grapes obtained in 2016 year

The average Weight of 100

Variety kg/vine weight of a grape berry
g g
E C E C E C
Feteasca neagra 3.59 4.23 94.43 93.65 161.6 173.6
Cabernet Sauvignon 3.04 3.14 84.75 93.57 135.2 140.1

Regarding the influence of experienced technological factors on the quality of

grape production, expressed by the sugar content and the acidity of the must, it is
found that the use of the conventional control system ensured the highest
concentrations of sugars in grapes. The sugar accumulation potential in the must, a
variety characteristic influenced by climatic factors during the grape maturation period,
was variable, with values between 194.6 g/L (conventional control system) and 190.8
g/L) and the Cabernet Sauvignon variety with limits of 206.3 g/L (conventional control
system) and 187.8 g/l (ecological control system), (Table 4).

Table 4
Quality wine harvest obtained in 2016

Sugars Total acidity

Variety
g/L g/L H2SO4
E C E C
Feteasca neagra 191 195 8.6 8.6
Cabernet Sauvignon 188 206 4.9 4.6

With regard to soil maintenance systems, the effects were differentiated,

depending on the specific conditions, the highest yields were obtained in the case of
soil maintenance by mulching with marc compost 3.83 kg/vine(ecologic control system)
and 4.23 kg/vine (conventional control system) in the Feteasca neagra variety, total
mulching with straws 3.76 kg/vine(ecologic control system) and 4.08 kg/vine
(conventional control system), black furrow 3.71 kg/vine(ecological control system) and
3.89 kg/vine (conventional control system), while the lowest yield was obtained in the
case of maintenance of soil by permanent herbage 3.59 kg/vine (ecological control
system) and 3.71 kg/vine (conventional control system), (Table 5) .

231
 Table 5
The influence of experienced technological factors on the production of grapes and its
qualities
a) Feteasca neagra
The Sugars Total
average acidity
Graduation kg/vine
weight of a
grape (g) g/L g/L H2SO4
E C E C E C E C
Black furrow 3.71 3.89 95.7 94.6 197.6 198.5 4.7 4.7
Total mulching
3.76 4.08 91.2 92.0 191.4 196.3 4.8 4.9
with straws
Partial mulching
with marc 3.83 4.23 90.1 95.6 187.8 186.2 4.9 4.6
compost
Permanent
3.59 3.71 94.4 92.0 190.9 199.8 4.8 4.7
herbage

b) Cabernet Sauvignon
The Sugars Total
average acidity
Graduation kg/vine
weight of a
grape (g) g/L g/L H2SO4
E C E C E C E C
Black furrow 3.00 3.07 90.5 86.9 196.8 195.4 8.4 8.4
Total mulching
3.05 3.08 86.9 91.9 192.1 190.6 8.5 8.6
with straws
Partial mulching
with marc 3.06 3.14 89.5 93.6 191.4 189.2 8.5 8.8
compost
Permanent
2.88 3.04 86.8 84.7 198.6 195.5 8.4 8.5
herbage

The highest grape sugar contents were obtained when the soil is maintained
by permanent herbage 190.9 g/L (ecological control system) and 199.8 g/L
(conventional control system) and the black furrow 197.6 g/l (organic control system)
and 198.5 g/l (conventional control system).
Regarding the influence of experimental technological factors on the degree of
attack of the main cryptogamic diseases: Plasmopara viticola, Uncinula necator and
Botrytis cinerea (Table 6), it was found that due to climatic conditions specific to the
2016 year (abundant rain and high hygroscopicity) showed the degree of attack on the
grape, with values ranging between 3.35% for the ecological control system and 2.87%
for the conventional control system at Feteasca neagra, and for the Cabernet
Sauvignon the variations were between 4.05% for the ecological control system and
2.98 % at the conventional control system.
The attack of Uncinula necator on grapes was 3.93% for the ecological control
system and 2.63% for the conventional control system for the Feteasca neagra variety
and between 2.26% for the ecological control system and 1.82 for the conventional
control system for the Cabernet Sauvignon variety. As for the Botrytis cinerea, it had
an attack rate of 1.97% (Feteasca neagra) and 2.26% (Cabernet Sauvignon) in the

232
ecological control system, and in the case of conventional control system the attack
rate was 1.27% (Feteasca neagra) and 1.82% (Cabernet Sauvignon).

Table 6
The degree of attack of the main diseases under the conditions of 2016

Plasmopara Uncinula Botrytis

Variety
viticola necator cinerea
E C E C E C
Feteasca neagra 3.35 2.87 3.93 2.63 1.97 1.27
Cabernet Sauvignon 4.05 2.98 4.51 2.13 2.26 1.82

Analyzing the sensory profile of the obtained wines it is found that the
experimental technological links influenced differently the 14 characteristics that make
up the sensory profile. In the case of Feteasca neagra, the best wine quality indices
were obtained in the conditions of soil maintenance as a black furrow and by total
mulching with straws (Figure 1).

Appearance
Harmony 6,0 Color intensity
5,0
Quality 4,0 Nuance
3,0
2,0
Onctuozity Olfactory quality
1,0
0,0
Extraction Olfactory intensity

Aggressiveness Gustative quality

Softness Gustative intensity

Astringency

Black furrow Total mulching with straws

Partial mulching with marc compost Permanent herbage

Figure 1. The influence of the soil maintenance system on the technological typicality
of the Feteasca neagra wines

In the case of the Cabernet Sauvignon variety, the best wine quality indices
were obtained under the conditions of soil maintenance as a black furrow and by total
mulching with straws (Figure 2).
In the case of Feteasca neagra and Cabernet Sauvignon wine, the best wine
quality indices were obtained under the conditions of soil maintenance as a black
furrow and by total mulching with straws and the ecological control system.

233
 Appearance
Harmony 6,0 Color intensity
5,0
Quality 4,0 Nuance
3,0
2,0
Onctuozity Olfactory quality
1,0
0,0
Extraction Olfactory intensity

Aggressiveness Gustative quality

Softness Gustative intensity

Astringency

Black furrow Total mulching with straws

Partial mulching with marc compost Permanent herbage

Figure 2. The influence of the soil maintenance system on the technological typicality
of the Feteasca neagra wines

CONCLUSIONS
With regard to the soil maintenance system, the highest sugar content in
grapes was obtained in the case of soil maintenance by permanent herbage and black
furrow.
The experimental data show that the permanent herbage exerts a negative
influence on the production of the vines, through the competition of the perennial herbs
in the use of water and mineral substances on the vine.
The use of the conventional control system has ensured the highest
concentrations of sugars in the grapes.

REFERENCES
Dejeu Liviu, Matei Petruţa, 1996 – Viticultura biologicǎ, Centrul Editorial Poligrafic
USAMV Bucureşti
Heidi Rosenthal Duminy, 2004 – Modern Ecological wine grape production in
South Africa, Masters diploma, pp. 1-63

234
 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
 Food produce process ing
UNIVERSITY OF CRAIO VA
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

NOROCEL - A NEW SEEDLESS GRAPE VARIETY CREATED IN ROMANIA

1 2* 1
Popa Camelia , Cichi Daniela Doloris , Ciobotea Cristina Magdalena
1
National Research and Development Institute for Biotechnology in Horticulture Stefanesti-Arges, Bucharest
Street - Pitesti, No.37,Stefanesti, Romania
2*
University of Craiova, Faculty of Horticulture, Department of Horticulture & Food Science,
Craiova, Romania
* Correspondence author. E-mail: [email protected]

Key words: seedless grapes, agrobiological descriptors

ABSTRACT
The present paper presents the results of the observations and determinations
regarding the evaluation of the fertility and productivity potential of a new grapevine variety,
Norocel, obtained at I.N.C.D.B.H. Stefanesti (Romania). Under the specific conditions of the
study years and the area investigated, the new variety Norocel proves superior to the Perlette
variety in terms of resistance to frost under some conditions of critical temperatures of – 17.5 ºC,
potential fertility and productivity of shoots, the average weight of a bunch, the production per
plant and qualitative potential.

INTRODUCTION
The ever-increasing of growing grape consumer requirements makes it
necessary to intensify research into the improvement of grapevine, with a focus on the
creating new Vinifera genotypes, with superior quality and productivity characteristics
with high ecological plasticity, high tolerance to diseases, pests, high resistance to
stress factors (Cichi Daniela Doloris et al. 2013, 2014; Ciobotea Cristina et al.2014;
Popa Camelia et al.2009; Rotaru Liliana et al.2008).
At present, the Romanian market is experiencing a shortage of seedless grape
varieties, but also with the existence of a limited number of varieties with extra season
and early maturation, but especially of those with complex biological resistance
(Damian Doina et al. 2006; Necula Cezarina et al.2014). In this context, at National
Research and Development Institute for Biotechnology in Horticulture Stefanesti -
Arges (I.N.C.D.B.H., Romania) was created a new variety of seedless grapevine,
Norocel, registered in the Official Catalog of the Crop Plants Varieties of Romania and
proposed for protection by patent in 2017.

MATERIAL AND METHODS

The researches were carried out between 2015 - 2016 in the experimental
fields of National Research and Development Institute for Biotechnology in Horticulture
Stefanesti - Arges (N.R.D.I.B.H., Romania), vineyard Stefanesti Arges, Muntenia and
Oltenia Hills. The climate favourability assessment was carried out as a synthesis of
several climatic factors with an impact on the bioproductive behaviour and quality of
the grapevine.

235
 Seedless variety Norocel (Figure1) was obtained by conducted hybridization of
varieties (Augusta x Askari) x Black Pearl, by Camelia Popa and Gheorghe Smaranda
at N. I. R. D. B. H. Stefanesti - Arges being enrolled in the Official Catalog of the Crop
Plants Varieties of Romania in 2017 and proposed for protection by patent in 2017.
To assess the bio productive and qualitative potential of the new seedless
grape variety Norocel and the Perlette variety - as the witness variety, a series of
agrobiological and technological descriptors were determined in accordance with the
O.I.V. (2002) methodology and I.C.V.V (1988).
To this end, observations and determinations were made on the following
agrobiological descriptors: proportion of fertile shoots (%), the absolute (c.f.a) and
relative fertility (c.f.r) coefficients, relative (I.p.r.) and absolute productivity (I.p.r) index.
The observations and determinations of the technological descriptors was concerned:
single bunch weight (g), production (kg/vine), sugar content of must (g/L) and total
acidity of must (g/L H2SO4).

Figure 1. Bunches of Norocel seedless table grapes cultivar

RESULTS AND DISCUSSIONS

As for the climatic resources in the viticultural experimental centre under study,
during the related period, they were analysed vs the requirements for the table grape
and seedless varieties. The main viticultural climatic characteristics during the period of
2015 and 2016 years are shown in Table 1.
The percentage of fertile shoots, as indicator of the real fructification potential
of varieties in a certain viticultural area, offers information on the specific genetic
potential of varieties, as well as on the good use of pedoclimatic resources and on the
value of the cultivation system, mostly depending on the photo-periodicity
characteristics, as well as on the nutrient reserves accumulation (Cichi Daniela et al.
2011).
The comparative study of the percentage of fertile shoots to the analysed
varieties reveals a number of differences between them. Thus, under the same agro-
pedoclimatic conditions of Stefanesti Arges vineyard, the Norocel variety (68%) is
characterized by superior fertility per plant, compared to seedless Perlette variety
(62%), due in particular to the viability of buds, in a higher percentage for the Norocel

236
variety (96%) compared to 31% as recorded to the Perlette variety under conditions of
absolute minimum temperatures of – 17.5 ° C recorded in 2016 (Figure 2).

Table 1
The climate characteristics during the vegetation period of years 2015 and 2016

Year
Viticultural climatic Indices
2015 2016
Average annual temperature (ºC) 12.5 12.00
Winkler thermal index (IW)- useful balance
1825 1806
sheet
Annual precipitations (∑ P mm) 804 797.4
∑ P (mm) during active vegetation period 478.6 519.6

Figure 2. Buds viability and proportion of fertile shoots

Fertility expressed by the values of fertility coefficients (c.f.r and c.f.a.)

highlights obvious differences both between varieties and between vineyards years
(Figure 3). The average values of the relative fertility coefficient were between 0.62
(Perlette) and 0.9 (Norocel), and those of the fertility coefficient absolute between 1.1
(Perlette) and 1.52 for the Norocel variety. We note the high fertility potential of the
Norocel variety as compared to that of the witness variety Perlette.
The productivity potential of the shoots to the studied varieties was expressed
by relative productivity indices (I.p.r) and absolute (I.p.a.).

Figure 3. Absolute (c.f.a) and relative fertility (c.f.r) coefficients

237
 Figure 4. Relative (I.p.r.) and absolute productivity index (I.p.a.)

Figure 5. Productive and qualitative characteristics

The relative productivity index, indicating how much a shoot produces in

average has, besides the value significance of the cultivation conditions, also the
function to determinate the load of buds per surface unit or per stock in order to
determine the planned production ( Cichi Daniela et al. 2011).
The relative productivity index was determined (I.p.r. = the average weight of a
bunch x c.f.r.) and the absolute productivity index was also calculated (I.p.a. = the
average weight of a grape x c.f.a.).
Very good results in terms of productivity have been recorded by the Norocel
variety (Figure 4), with average values of productivity indices between 479 (2016) and
559 (2015), for the absolute productivity index (I.p.a) and between 322 and 363 for the
relative productivity index (I.p.r.).
The evolution of climatic factors during the grape maturity has a decisive
influence on the process of grape maturation and finally on the quality of the wine-
produced wine production.
Depending on the climatic supply characteristic of the wine years 2015-2016,
the full maturation of the Norocel variety was achieved from 28.08-01.09 with 4-5 days
later than the Perlette variety.
Regarding the qualitative potential of the two seedless genotypes, it was found
that the highest sugar accumulation potential (Figure 5) was found in the Norocel
variety (140 g / L and 169 g / L must) in the context of in the context of a total acidity of
3.79-3.89 g / l of H2 SO4.

238
 Figure 6. Average production per vine

We appreciate that although the two genotypes reach full maturation

approximately in the same calendar period, the Norocel variety has a high potential for
accumulation in sugars, which ultimately results in a balance of the glucose-acidic
index and the taste qualities of this cultivar.
The Norocel cultivar is also distinguished by superior production / plant
compared to the Perlette variety (figure 6), the average production being 5.3 kg / plant
in 2015 and 8.9 kg / plant in 2016.
This is also explained by the higher average weight of the berry (3.4 - 3.8 g)
and the bunch (370 - 431 g) recorded to the Norocel variety compared to the seedless
Perlette variety ( Figure 5).
CONCLUSIONS
Under the specific conditions of the study years and of the investigated area,
the new Norocel grape variety is superior to the Perlette variety in terms of resistance
to frost under critical temperatures of - 17,5 ºC, the potential fertility and productivity of
the shoots, the average weight of a bunch, production/plant and qualitative potential
(sugar g/L).
In this context, the new seedless - Norocel grape variety is recommended to
be introduced into culture in very favourable and favourable areas for table grape
varieties in the south of the country, in order to evaluate its behaviour and its
expansion into culture.

REFERENCES
Cichi Daniela Doloris, Monica Vintilescu, Giugea N. Costea D.C., Camelia Popa,
2011, Agrobiological and technological potential of sometable grape varieties in the
Banu Maracine vineyard, Annals of the University of Craiova- Biology, Horticulture,
Food Produce Processing Technology, Environmental Engineering, vol. XVI (LII)/
2011, 87-92 p.
Cichi Daniela Doloris, Popa Camelia, Ciobotea Cristina Magdalena, Stoica Felicia,
Costea D.C., 2013, Assortment of table grape varieties and geographical distribution in
viticulture areas of development regions South Muntenia and South-west Oltenia of
Romania, Annals of the University of Craiova- Biology, Horticulture, Food Produce
Processing Technology, Environmental Engineering, vol. XVIII (LIV), p. 89-100
Ciobotea Cristina Magdalena, Popa Camelia, Cichi Daniela Doloris, Giugea N.,
2014, Comparative study regarding the agrobiological and technological characteristics
of some hybrid elites for table grapes obtained from self-pollination Victoria x Victoria ,

239
Annals of the University of Craiova- Biology, Horticulture, Food Produce Processing
Technology, Environmental Engineering, vol. XIX (LV), 113-118p
Damian Doina, Calistru Gh., Savin C., Vasile Ancuţa, Zaldea Gabi, 2006,
Valoarea agrobiologică şi tehnologică a unor soiuri noi şi clone de viţă de vie create la
SCDVV Iaşi, Lucrări ştiinţifice, seria Horticultură, vol. 49, U.S.A.M.V. Iaşi.
Necula Cezarina, C.Stirbu, Popa Camelia, 2014, Elite hybrid of perspective for
table grapes obtained in INCDBH Stefanesti, GeoConference on Nano, Bio And
Green–Technologies For A Sustainable Future, SGEM 2014 Conference Proceedings,
ISBN 978-619-7105-20-9/ISSN 1314-2704, June 19-25, 2014, Vol. 1, 359-364 pp
Popa C., Necula C., Cichi D., Giugea N., 2009, Argessis and Golden Stefanesti
new varieties for table grapes with high biologycal strenght- P.I.06- 32 Congress of
vine and wine, June 28th- July 3rd 2009-Zagreb-Croatia – Edition Naklada
Rotaru Liliana, Mustea M., Zamfir C., Cotea V.V., Vasile Ancuţa, 2008, New
vinifera creations for table grapes in the restrictive conditions of culture in the North-
Eastern area of Romania, XXXIth – World Congres of Vine and Wine OIV, Verona, 15-
20 june.
***Buletinul I.C.V.V., nr. 7 (2/1988), ICVV Valea Călugărească
*** 2ndEdition of the OIV Descriptor list for grape varieties and Vitis species, 2002,
OIV, Paris

240
 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
 Food produce process ing
UNIVERSITY OF CRAIO VA
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

RESEARCH REGARDING THE EFFECT OF GROWTH

REGULATORS OVER THE IN VITRO MORPHOGENETIC
REACTION AT SALVIA OFFICINALIS
1* 1
Radomir Ana-Maria , Ciobotea Cristina-Magdalena
1
National Research and Development Institute for Biotechnology in Horticulture, Stefanesti,
* Correspondence author. E-mail: [email protected]

Key words: Salvia officinalis, “in vitro”culture, explants, microshoots, growth regulators

ABSTRACT
The goal of this work was to develop a protocol for obtaining a high regeneration rate
by testing the response of different explants of Salvia officinalis species on different culture
media. Results showed that explants of apexes stems represent optimal source of inoculums.
Research in the evaluation of morphogenetic capacity of explants on different nutrient
regeneration formulas have shown that the most effective hormonal balance in the generation of
shoots was 3 mg/l BAP / 1 mg/l ANA, in which case the rate of multiplication was 5,25
microshoots/explant. Shoots were multiplied by subculture on the same medium. The highest
percentage of rooting (94.2%) was registered in the case of supplementing the nutrient medium
with NAA in a concentration of 0.6 mg/l. The rooted plantlets were acclimatized and successfully
fortified in pots registering a survival rate of 87%.

INTRODUCTION
Salvia officinalis (garden sage) is a perennial subshrub from the family
Lamiaceae. This plant originates from the Mediterranean region and is widely used in
pharmaceutical and food industries as well as in cosmetics (Khan et al., 2010; Garcia
et al., 2013). The leaf extract of the plant possesses antibacterial, antifungal, antiviral,
anti-inflammatory, antitumor and hypoglycemic activity (Longaray Delamare et al.,
2007; Keshavarz et al., 2011). The pharmacology action of sage is a result of the
presence of essential oil, di- and triterpenes, phenolic acids, flavonoids and tannins
(Azimova and Glushenkova, 2012; Kontogianni et al., 2013). There are several studies
concerning micropropagation of Salvia officinalis using different explants: shoot tips
(Grzegorczyk and Wysokinska, 2008), nodal segments (Gostin, 2008), axillary and
apical buds (Wielgus et al., 2011). However, some of the serious limitations in the
above mentioned protocols were low propagation frequency, inconsistent and less
number of shoots and occurrence of hyperhydricity.
The aim of this study was to develop an efficient method for micropropagation
of Salvia officinalis.

MATERIALS AND METHODS

The biological material derived from Salvia officinalis species has been
multiplied in vitro to yield neoplantlets regenerated by tissue culture.

241
 Explants used to initiate tissue culture was represented by nodes and apexes
stems. The defoliated shoots were first washed in tap water and were sterilized in 6%
HgCl2 for 10 minutes, after which 3 rinses were performed in sterile distilled water.
After sterilization of plant material, the explants were sampled and inoculated
on MS culture medium without growth regulators. As a carbon source was used
dextrose (40 g/l) and as a source of iron was used NaFeEDTA (32 g/l). For
solidification of the culture medium was used agar (7 g/l).
The culture media were sterilized by autoclaving at 120°C temperature for 20
minutes. Before autoclaving, the pH registered in a culture medium was adjusted to
5.6-5.8.
The reactivity of explants was evaluated after a month of culture.
In order to culture proliferation, the explants by a node taken from microshoots
obtained in the initiation cultures phase were inoculated on MS culture medium
supplemented with various concentrations and combinations of BAP, K and ANA: 3
mg/l BAP / 0,5 mg/l ANA (V.1.), 2 mg/l BAP (V.2.), 1 mg/l BAP (V.3.), 1 mg/l K / 0,1
mg/l ANA (V.4.), 3 mg/l BAP / 1,0 mg/l ANA (V.5.). The cultures were transferred each 4
weeks on fresh media.
The differentiated microshoots, with a length of 3-4 cm, were detached from the
culture, individualized and cultivated on rooting medium, represented by a variation of the
MS basal medium with mineral salts reduced to half in which the auxin type varied (ANA -
V.1., AIA - V.2., IBA - V.3.) and was kept constant its concentration (0.6 mg / l).
For growth, multiplication and rooting of explants have ensured in the growing
room controlled conditions of temperature (22-24oC), photoperiod (16 hours) and light
intensity (2 500 lx).
The in vitro rooted plants were transferred ex vitro in order to be
acclimatization at their natural environment.
For the regeneration stage of the cultures, the monitored parameter was the
multiplication rate calculated as the number of differentiated shoots on the explant. The
percentage of rooting the shoots in vitro was estimated as the ratio of the number of
shoots to which the rhizogenesis induction process occurred and the total number of
shoots placed on the rooting substrate. The acclimatization percentage of in vitro
regenerated plant was calculated as the ratio between the number of acclimatized
plants viable and total number of in vitro regenerated plants.

RESULTS AND DISCUTIONS

Research on in vitro multiplication of the species Salvia officinalis have showed
the complexity of the process and revealed the influence of biotic (type of explant) and
abiotic (nutritive substrate) factors on the different phases of initiation, regeneration,
rooting and acclimatization of regenerated plants. To establish a reproductible protocol
for in vitro multiplication was necessary the selection of explant and of optimal culture
medium to induce a higher number of shoots.
The observations made during the initiation stage of in vitro culture revealed
that the inclusion of sage in this culture system does not pose particular problems, the
use of calcium hypochlorite (a 6% solution for 10 minutes) for the sterilization of the
explants proved to be efficient. The use of basal medium MS without growth hormones
favored the process of shoots development and the production of neoplantlets which
were then used to test the morphogenetic reaction of sage explants on different
hormonal formulations of the MS medium (Figure 1).
In comparative analyzes aimed morphogenetic competence of explants from
different origins (nodes and apexes stems), the results obtained showed that both
types of explants analyzed have the ability to regenerate shoots under in vitro

242
conditions, but the highest morphogenetic potential was observed in the case of
apexes stems (Figure 2).

Figure 1. Minishoots inoculated on MS Figure 2. The morphogenetic response

medium without growth regulators of the apexes stems explants, after 4
weeks from initiation of in vitro culture

In order to proliferate the culture, the explants of a node taken from the
microshoots obtained in the initiation phase of the cultures were inoculated on the MS
culture medium supplemented with different concentrations and combinations of
phytohormones.
The results registered in shoots regeneration experiments at sage reflected the
important role that it has hormonal balance in overall of the factors that determine the
expression of regenerative potential in the in vitro culture of the explants, a conclusive
evidence in this regard being different effects of the same basal medium supplemented
with various combinations and concentrations of growth hormones.
From the five variants of culture medium tested, two (V1 and V5) led to a better
morphogenetic response, two (V2 and V3) allowed induction of the regenerative
processes but to a lesser extent, and one (V4) did not stimulate the production of
shoots. It is well known that directing of the morphogenetic processes to the
development of shoots directly from the explant can be accomplished by
supplementing the basic nutrient media with different types of cytokinins. In our
experiments, the research was performed on three of the most important cytokinins,
namely BAP, NAA and K. The results obtained showed that among the three types of
cytokines used, BAP was the most effective in the shoot regeneration. Each of the four
variants of nutrient media which allowed the induction of regenerative processes was
characterized by the presence of BAP (V2 and V3) or BAP in combination with ANA
(V1 and V5). The most appropriate hormonal balance for this purpose was found to be
3 mg / l BAP / 1 mg / l ANA (V5), in which case the value of the multiplication rate was
5.25 minishoots / explant. Replacement of BAP with K (V4) did not stimulate shoots
regeneration, the multiplication rate being 0 (Figure 3).
After approximately 30 days, the shoots obtained were transferred to fresh
medium that supported the regenerative processes by determining a good proliferation
of shoots (Figure 4).

243
 Figure 3. The influence of growth Figure 4. Biological material multiplied
hormone combination and concentration in vitro
on in vitro multiplication rate to the
species Salvia officinalis

From the qualitative point of view, the biological material resulting from the
regeneration of explants had a normal morphology, without vitrification aspects,
necrosis or callus differentiation.
Differentiated microshoots were detached from the culture, individualized and
cultured on rooting medium. Knowing the beneficial effect of auxins on the efficiency of
the rhizogenesis process, three variants of the rooting medium were tested in which
the auxin type (ANA, AIA, IBA) varied, and its concentration was maintained constant
(0.6 mg /l). From the three variants tested, the highest rate of rooting (94.2%) was
registered in the V1 variant, which is characterized by the presence of the auxin NAA
at a concentration of 0.6 mg / l. Variants V2 and V3, although lead to the rooting of the
shoots in a fairly high percentage - 88.5%, respectively 72.4%, failed to reach a value
of 94.2% (rate obtained for V1 variant) (Figures 5 and 6).

Figure 5. In vitro rooting capacity at Figure 6. Rooting of Salvia officinalis

Salvia officinalis depending on the type microshoots on ½ MS supplemented
of auxin with 0.6 mg / l ANA
In vitro rooted plants were transferred ex vitro in order to accommodate to
septic life conditions. Their acclimatization has been successful, plants having a well
developed radicular system, capable of further supporting growth and development
process.
The best results with regard to acclimatization of Salvia officinalis plants has
been obtained when have been used pills of peat type Jiffy (87% acclimated plants).
When using the mixture of peat, perlite and manure in proportion of 2: 1: 1, the
percentage of acclimatized plants was lower (75%), this substrate however ensures a
good fortification of acclimated plants (Figure 7).
244
 a b

Figure 7. Acclimatization of in vitro regenerated plants: a) peat, manure, perlite

mixture; b) pills of peat

After the acclimatization and fortification at pots, the plants were transplanted
to greenhouse for further their growth and development (Figure 8). Mention the fact
that plants obtained by in vitro propagation have preserved morphological
characteristics of donor plant.

a b

Figure 8. Salvia officinalis plants fortified in pots (a) and on soil (b)

We conclude that results obtained throughout in vitro regeneration phases,

confirming preliminary studies and certify that for Salvia officinalis species, cloning by
tissue culture is viable. Micropropagation technique represents an effective alternative
to propagating for species under study, in order to create a permanent source to
extract of some secondary metabolites for pharmaceutical industry.

CONCLUSIONS
 The results obtained with regard to the morphogenetic competence of
explants from different origins (nodes and apexes stems) revealed that the apexes
stems have the highest morphogenetic potential.
 Of the three types of cytokinins used, the most effective in the production of
shoots was BAP. Replacing BAP with K did not stimulate shoot regeneration.
 The highest percentage of rooting (94.2%) was recorded in the case of
supplementation of the nutritional medium with ANA at a concentration of 0.6 mg/l.
 The yield on acclimatization was influenced by nutritive support used. The
best results were obtained by using peat pills as substrate nutrient which ensured a

245
percentage of 87% acclimatized plants. We recommend the mixture of peat, manure
and perlite in a proportion of 2:1:1 for the fortification of vitroplants in pots.
 On the basis of the results obtained was developed an original,
reproducible and efficient in vitro multiplication protocol to the sage.

REFERENCES
Azimova SS, Glushenkova AI, 2012. Salvia officinalis L. In Lipids, Lipophilic
Components and Essential Oils from Plant Sources. Springer London. pp. 494–494.
Garcia CSC, Ely MR, Wasum RA, Zoppa BCDA, Wolhheim C, Neves GÂ, Angeli
VW, Souza KCBD, 2013. Assessment of Salvia officinalis (L.) hydroalcoholic extract for
possible use in cosmetic formulation as inhibitor of pathogens in the skin. Revista de
Ciências Farmacêuticas Básica e Aplicada, 33(4): 509–514.
Gostin I, 2008. Effects of different Plant Hormones on Salvia officinalis, cultivated
in vitro, Int J Bot, 4 (4): 430–436.
Grzegorczyk I, Wysokińska H, 2008. Liquid shoot culture of Salvia officinalis L. for
micropropagation and production of antioxidant compounds; effect of triacontanol. Acta
Societatis Botanicorum Poloniae, 77(2): 99–104.
Keshavarz M, Bidmeshkipour A, Mostafaie A, Mansouri K, Mohammadi-Motlagh
HR, 2011. Anti tumor activity of Salvia officinalis is due to its anti-angiogenic, anti-
migratory and anti-proliferative effects. Cell J, 12: 477–482.
Khan IA, Abourashed EA, 2010. Leung‟s Encyclopedia of Common Natural
Ingredients: Used in Food, Drugs and Cosmetics. (Hoboken, Ed.) (3rd ed., p. 845).
N.J.: John Wiley & Sons Inc.
Kontogianni VG, Tomic G, Nikolic I, Nerantzaki AA, Sayyad N, Stosic-Grujicic S,
Stojanovic I, Gerothanassis IP, Tzakos AG, 2013. Phytochemical profile of Rosmarinus
officinalis and Salvia officinalis extracts and correlation to their antioxidant and anti-
proliferative activity. Food Chem, 136(1): 120–129.
Longaray Delamare AP, Moschen-Pistorello IT, Artico L, Atti-Serafini L,
Echeverrigaray S, 2007. Antibacterial activity of the essential oils of Salvia officinalis L.
and Salvia triloba L. cultivated in South Brazil Food Chem, 100(2): 603–608.
Wielgus K, Luwanska A, Szalata M, Mielcarek S, Gryszczynska A, Lipinski D,
Slomski R, 2011. Phytochemical estimation of Sage (Salvia officinalis L.) cultivated in
vitro-flavonoids and phenolic acids. Acta Fytotechnica et Zootechnica, Special Number
Nitra, 8–11.

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 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
 Food produce process ing
UNIVERSITY OF CRAIO VA
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

RESEARCH ON DEPENDENCE OF GRAPE PRODUCTION OF TEMPERATURE IN THE

REST PERIOD AT SOME WHITE WINE GRAPES IN THE SANDY SOILS FROM SOUTH-
WESTERN OLTENIA
Iulian Rățoi1*
1-
Research – Development Center for Plant on Sands Dabuleni, Victoria Street, Dabuleni.
*
Correspondence author: e-mail: [email protected]

Key words: vine, negative temperature, abundant rainfall

ABSTRACT
Determinations were performed over the 2012-2016 period, on a plantation with the
classic form of leadership (low). The state of vegetation and vine production is located
permanently subject to weather conditions. In the favorable climatic conditions (2013 year), all
varieties have been expressed at the level of genetic potential and of the degree of adaptability.
The highest level of production was achieved at the noble variety Selena with 35.9 t/ha, followed
by the resistant variety Brumăriu (34.8 t/ha). Analysis of grapes average production, conducted
in the first 4 years of fruiting, highlights the special ability of fruits variety with ,,blood of hybrid”
Brumăriu, 18 t/ha, a superior production of noble varieties Selena and Blasius which achived a
grapes average production of 13.9 and respectively 13.8 t/ha.

INTRODUCTION
Climatic factors, together with a edaphic factors, are limiting factors for vines.
Climatic factors as well as harmful minimum temperatures from winter and spring frosts,
sometimes, determines the utmost grape production (Țârdea C., Dejeu L., 1995), even when
the taught elements of fruiting, arranged at the base of the vine, are protected by earth,
because and the practiced cultural measures are extremely important in minimizing the
damage caused by harmful minimum temperatures at vine (Robert G.E., 2000). In some
years the damage can be total when intracellular ice crystals are formed, affecting the main
buds and wood annual and multiannual even. The risk of freezing at the base of the vine is
greater than to height of 100 cm, it being reduced by up to 0.36°C every 10 cm above the
ground level (Sugar, D. R. et. al., 2003).
In current circumstances, when they manifest changes or climate disruption is
necessary to improve the existing product range, with varieties showing superior quantitative
and qualitative traits, of the current range, and adapt to sandy soil from south-western area
(Vlădoianu Em., 1979; Vlădoianu Em., 1981; Anca Onache et al., 2008).

MATERIAL AND METHODS

The experience was founded in 2010. In 2012 year the plantation was found in the
third year after planting, and in 2013 year was the first year of fruiting.
Were plannted the varieties following: Columna, Donaris, Fetească albă 1 Od.,
Fetească regală 21 Bl, Pinot gris, Riesling italian, Riesling de Rhin, Neuburger, Chasselas
d’ore, Muscat Ottonel, Sauvignon blanc, Alb aromat, Grasă de Cotnari, Brumăriu, Blasius,
Sauvignon, Selena.

247
 Planting density is 3787 vines/hectare, density which results from planting distances
of 2.2 / 1.2 m.
During 2012-2016 period, the climatic conditions were unfavorable for vine in winter
of 2012, 2015, 2016 years, and during the vegetation period in 2014 year due to abundant
rainfall (640.7 mm in vegetative period) spreaded over a number of days, 74 of 183 total
(48.5%), but and of phenomenon of dew shown daily throughout the year (table 1). In the 3
years previously named temperatures recorded were below the limit of resistance for buds
and other organs of the vine, -24.30C in 2012 year, -25.10C in 2015 year and -23.40C in
2016 year.
At these temperatures were left only one part of the buds whith were placed in the
snow, the thickness of which was 25-30 cm. In the five years in which conducted the study,
only one was normal, in terms of climate conditions for vine, 2013 year, when the absolute
minimum temperature of -18.10C, was not affected the viability of main buds.
In 2014 year, allthouth the vine did not suffer due to low temperatures, grape
production was affected by the large amount of precipitation, 994 mm annual, over the
average multiannual of rainfall.
In years when vines were affected by frost, cutting short fruiting was performed in
short elements, of 2-3 eyes, as the existing buds above the layer snow were destroyed in
fully, and with these annual and multiannual and wood. In these conditions we have secured
fruit load specific to each variety.
Were effectuated observations and determinations experimentale on grape
production and quality (weight of 100 grapes, the content of total sugars and total titratable
acidity).
Table 1
The main climatic elements from 2012-2016 period
0
Year Temperature ( C) Rainfall (mm)
Minimum Maximum Annual April- Multiannual Snow
September layer (cm)
period
2012 -24.3 42.6 383.5 230.8 578.5 25
2013 -18.1 38.4 451.5 307.8 588.4 15
2014 -14.1 37.6 994 640.7 546.8 25
2015 -25.1 39.2 748.9 398 554.9 30
2016 -23.4 38 748.9 297.4 554.9 30

RESULTS AND DISCUSSIONS

Negative minimum temperatures registered during the rest period of the 2012,
2015 and 2016 years and abundant rainfalls of 2014 year, were constituted in harmful
factors for vines in this area. In 2012 year, grape production was lower due to the fact
that the vine has been in the third year after planting, in the last year of formation
(table 2). This demonstrates that not all varieties have the same precocity in the bear.
In 2013 year, the favorable climate has made all the varieties to express with
the genetic potential and the degree of adaptability, specific to each variety individually
for the first year of fruit, in conditions of sandy soils area from south-western Oltenia.
The highest level of production of grapes was done at Selena noble variety, with 35.9
t/ha, followed a short distance from resistant variety Brumăriu (34.8 t/ha). From noble
varieties were also noted Blasius (30.0 t/ha), Alb aromat (29.2 t/ha) and Chasselas
dʹoreˊ (27.6 t/ha). With a production of grapes over 20 t/ha, were found varieties
Fetească albă Cl. 1 Od. (21.2 t/ha) and Fetească regală Cl. 21 Bl. (21.2 t/ha). Harder
adaptable in this area and with lower genetic potential were Muscat Ottonel (11.7
t/ha), Neuburger (11.3 t/ha) and Donaris (10.2 t/ha).

248
 Table 2
Yield production (t/ha) of some varieties with grapes for white wine in 2012-2016
period

Variety Year 2013-2016

2012 2013 2014 2015 2016 Average
Riesling italian 0.7 16.7 7.6 2.3 16.3 10.7
Columna 0.7 12.9 7.2 2.9 11.7 8.7
Donaris 0.7 10.2 3.0 0.9 11.3 6.3
Fetească albă Cl. 1 Od. 0.4 21.2 7.9 3.6 6.8 9.9
Fetească regală Cl. 21 Bl. 3.9 21.2 10.2 7.5 10.6 12.4
Pinot gris 0.4 12.5 0 1.8 7.2 5.4
Riesling de Rhin 0.7 15.5 3,8 2.4 7.6 7.3
Neuburger 0.4 11.3 0 3.4 6.4 5.3
Chasselas d'oré 3.0 27.6 0 4.9 9.1 10.4
Muscat Ottonel 0.7 11.7 3.4 1.4 4.5 5.2
Sauvignon blanc 0.7 21.2 0 2.9 8.3 8.1
Alb aromat 4.2 29.2 8.3 0.5 7.2 11.3
Grasă de Cotnari 1.1 19.7 0 0.6 7.6 7.0
Brumăriu 4.2 34.8 10.6 4.9 21.9 18.0
Blasius 4.5 30.0 8.7 8.9 7.6 13.8
Sauvignon 0.7 14.0 3.4 3.6 10.6 7.9
Selena 1.5 35.9 6.4 6.8 6.4 13.9
In 2014 year, temperatures were favorable for dormant vines, but the abundant
rainfall during the growing season increased vulnerability against the major fungal diseases
of plants and increased their virulence, so that the pesticide treatments carried out have failed
to halt their work, or if the moment was stopped the next rain or even dew diseases were
reactivated.
Some varieties have shown greater tolerance and achieved grape production 8-10 t/ha,
Brumariu variety (10.6 t/ha), which is part of resistant varieties, but also noble varieties
Fetească regală Cl. 21 Bl. (10.2 t/ha), Blasius (8.7 t/ha) and Alb aromat (8.3 t/ha). At some
varieties, more sensitive grape production was compromised, example varieties Pinot gris,
Neuburger, Chasselas d’ore, Sauvignon blanc, Grasă de Cotnari. The adverse effect of
minimum temperatures on vines in 2015 year, was shown amid of vines diseases by 2014
year. So that, in 2015 year, grapes production achieved by studied varieties was extremely
low. Blasius, Fetească regală Cl. 21 Bl. and Selena varieties who realized the production of
grape between 8.9 and 6.8 t/ha. On the other varieties of grape yields realized was
insignificant.
In autumn of 2015 year, was protected with earth 2-3 strings from each vine, so
that, the negative effect of harmful minimum temperatures in winter 2015-2016, has declined
more were less depending on the variety. The grapes production achieved was over 6.4 t/ha,
culminating with resistant variety Brumăriu which achieved a grapes production of 21.9 t/ha.
Noble variety was highlighted with a Riesling italian variety, with a grape production of 16.3
t/ha.
Table 3
Weight of 100 grapes (g) in harvesting moment of some varieties with grapes for
white wine in 2012-2016 period
Variety Year Multiannual Average
2012 2013 2014 2015 2016 interval
Riesling italian 133 166 180 204 150 133-204 166
Columna 180 155 258 256 205 155-258 211
Donaris 182 178 244 267 263 178-267 227
Fetească albă Cl. 1 Od. 126 181 142 163 147 147-181 152
Fetească regală Cl. 21 Bl. 137 172 182 170 205 137-205 173
Pinot gris 92 140 156 127 125 92-156 128

249
Riesling de Rhin 126 152 156 159 167 126-167 160
Neuburger 154 209 202 259 205 154-259 206
Chasselas d'oré 230 211 196 316 222 196-316 235
Muscat Ottonel 146 224 196 275 183 146-275 205
Sauvignon blanc 133 171 158 192 167 133-192 164
Alb aromat 313 282 288 368 354 282-354 321
Grasă de Cotnari 208 276 276 248 333 208-333 268
Brumăriu 159 217 180 209 189 159-217 191
Blasius 218 288 392 420 395 218-420 343
Sauvignon 121 174 180 190 168 121-190 167
Selena 149 218 196 263 221 149-263 209
They also noted varieties Columna (11.7 t/ha), Donaris (11.3 t/ha), Fetească
regală Cl. 21 Bl. and Sauvignon with 10.6 t/ha, production of grapes each. Analysis of
grape average production, realised in the first 4 years of fruits, highlights the special
ability of fruiting for variety with ,,blood hybrid” Brumăriu, 18 t/ha, a superior production
of noble varrieties Selena and Blasius which achived a grapes average production of
13.9 and respectively 13.8 t/ha.
Table 4
Sugar content (g/l) in the harvesting moment at some varieties with grapes for white
wine in 2012-2016 period
Variety Year Multiannual
2012 2013 2014 2015 2016 interval
Riesling italian 209 208 197 183 198 183-209
Columna 188 166 177 165 155 155-188
Donaris 185 166 211 206 180 166-211
Fetească albă Cl. 1 Od. 184 188 163 174 178 163-188
Fetească regală Cl. 21 Bl. 187 177 174 168 187 168-187
Pinot gris 199 220 182 167 189 167-220
Riesling de Rhin 192 206 187 171 177 171-206
Neuburger 200 217 183 181 189 181-217
Chasselas d'oré 185 179 142 156 167 142-185
Muscat Ottonel 224 222 186 194 182 182-224
Sauvignon blanc 230 215 178 188 186 160-230
Alb aromat 190 188 160 168 174 160-190
Grasă de Cotnari 185 182 167 152 176 152-185
Brumăriu 177 179 174 183 193 174-193
Blasius 203 172 159 178 175 159-203
Sauvignon 196 227 191 186 184 184-227
Selena 186 222 175 180 172 172-222
With chances of reaching the assortment of sandy soils is Fetească regală Cl.
21 Bl., which conducted a grape average production of 12.4 t/ha. Weight of 100 grapes
(g), is an indicator of quality vine. This varies depending on the variety, the load of
grapes per vine, the way it was maintained plantation, climate, etc. (table 3). Overall
varieties of wine have grape berries small, weighing between 1 to 2.3 g/grape, or
medium grape who weight is 2.4 to 5 g. In the case of varieties studied in this period,
the majority have small-berry grapes, namely 13 at number and only 4 have over 2.3
g/grape (Blasius, 3.4 g; Alb aromat, 3.2 g; Grasă de Cotnari, 2.7 g and Chasselas
d'oré, 2.4 g).
The content of total sugars in the grapes in the time of harvesting, is a
characteristic of the variety, but it depends on edaphic and climatic factors, the level of
production and the health of the grapes, etc., and oscillated between broad limits from
one year to the next (table 4).
The studied varieties have a high potential for accumulation of sugars, over 220 g/l,
thus ensuring a wine with more than 12.5 vol.% alcohol, in certain years, example
Sauvignon blanc (230 g/l), Sauvignon (227 g/l), Muscat Ottonel (224 g/l), Selena (222
g/l).
250
 Those varieties have accumulated the minimum amount of total sugars as
follows: Sauvignon blanc (160 g/l), Sauvignon (184 g/l), Muscat Ottonel (182 g/l),
Selena (172 g/l). In years with optimal conditions for accumulation of sugars, no one
variety has not accumulated under 185 g/l.
Fluctuating levels of sugars of grapes from one year to another, not offer a guarantee
to obtain a constant quality for wine permanently, so that, in this area you can get top-
quality wines only in certain years.

Table 5
Total titrable acidity (g/l H2SO4) in the harvesting moment at some varieties with
grapes for white wine in 2012-2016 period

Variety Year Multiannual

2012 2013 2014 2015 2016 interval
Riesling italian 3,1 4,2 5,2 4,1 5,0 3,1-5,2
Columna 3,3 3,8 4 2,7 4,5 2,7-4,5
Donaris 2,4 3,9 4,5 3,6 3,5 2,4-4,5
Fetească albă Cl. 1 Od. 2,5 3,4 4,2 2,7 4,0 2,5-4,2
Fetească regală Cl. 21 Bl. 2,6 3,8 4,4 5,0 5,6 2,6-5,6
Pinot gris 4 4,6 4,6 5,3 4,7 4-5,3
Riesling de Rhin 3,6 4,6 4,6 5,7 5,2 3,6-5,7
Neuburger 3,9 4,2 5,6 4,2 5,8 3,9-5,8
Chasselas d'oré 2,7 3,2 4,1 2,5 4,5 2,5-4,5
Muscat Ottonel 2,5 2,9 3,5 2,5 3,5 2,5-3,5
Sauvignon blanc 3,9 4,2 4,4 5,3 5,6 3,9-5,6
Alb aromat 3,1 3 3,3 2,7 3,5 2,7-3,5
Grasă de Cotnari 4 4,2 5 4,8 5,7 4-5,7
Brumăriu 4,2 4,1 2,6 5,3 4,5 2,6-5,3
Blasius 4,1 4,8 4,6 4,3 5,7 4,1-5,7
Sauvignon 3,8 4,2 4,9 4,0 5,8 3,8-5,8
Selena 3,5 4,2 4,5 3,0 3,5 3-4,5

Total titrable acidity, like the other two indicators of the quality of the wines
listed above, has a level fluctuating from one variety to another, from one year to the
next (table 5). In the area of sandy soils of South-western Oltenia, the value of this is
less than in other areas, for variety, due to much higher temperatures (above 35oC),
from July to August, when a good part of the day the main plant physiological
processes are blocked. The value of total titrable acidity is influenced by the health of
the grapes.
In certain years, some varieties can record total titrable acidity values between
4.5-5.5 g/l H2SO4, which conduce to balanced taste. In years when the acidity values
are around 3-3.5 g/l H2SO4, obtained fade wines, and in these situations it is necessary
to correct this value to improve the taste.

CONCLUSIONS
State of vegetation and vine production is constantly under incidence of
climatic conditions. Harmful minimum temperatures to the rest period, but other climate
elements, outside the optimal manifested during the growing season, producing
significant damage on vegetative and generative organs, and thus the production of
the year or even the next few years.
From the five years analyzed (2012-2016), optimal conditions for vines on
sandy soils were just in one year. In 3 years of these, negative minimum temperatures
recorded below the resistance of vine organs, they were harmful, and in a year of
abundant rains during the growing season, spread over a number of days (48.5% and
251
dew phenomenon manifested daily, in this period, were established in factors
unfavorable for this crop, which affected grape production and quality.
In favorable weather conditions, all varieties are expressed at genetic potential
and degree of adaptability. The highest production was achieved at Selena noble
variety, with 35.9 t/ha, followed by resistant variety Brumăriu (34.8 t/ha). They also
noted the noble varieties Blasius (30.0 t/ha), Alb aromat (29.2 t/ha) and Chasselas
dʹoreˊ (27.6 t/ha). Harder adaptable in this area and with lower genetic potential were
Muscat Ottonel (11.7 t/ha), Neuburger (11.3 t/ha) și Donaris (10.2 t/ha).
In 2014 year, the rainfall during the growing season has increased vulnerability
against the major fungal diseases of plants and reduced effectiveness of substances
used in plant protection treatments, so that at some varieties that year production was
compromised, example Pinot gris, Neuburger, Chasselas d’ore, Sauvignon blanc,
Grasă de Cotnari.
In years with harmful minimum temperatures (2016 year), protecting the earth
2-3 wood shoots from each vine, led to reduced negative effect. Grape production was
achieved over 6.4 t/ha, culminating with resistant variety Brumăriu that has achieved
grape production of 21.9 t/ha. Noble varieties was highlighted with a variety Riesling
italian grape production of 16.3 t/ha. Analysis of grape production average, conducted
in the first 4 years of fruits, highlights the special ability of fruiting for variety Brumariu
with ,,blood hybrid”, 18 t/ha, a superior production for noble varieties Selena and
Blasius which achieved average grapes production of 13.9 and 13.8 t/ha, respectively.
With chances of reaching the assortment of sandy soils is Fetească regală Cl. 21 Bl.,
which conducted a grape average production of 12.4 t/ha. The main quality indicators
(weight of 100 grapes, content of total sugar and total titrable acidity), it is a
characteristic of variety but also depends on climatic and edaphic conditions, and the
applied technology. In certain years the some varieties as well as Sauvignon blanc
(230 g/l), Sauvignon (227 g/l), Muscat Ottonel (224 g/l), Selena (222 g/l), realized
wines over 12.5 vol.% alcohol and an total acidity of 4.5-5 g/l H2SO4.
ACKNOWLEDGMENT
The paper is a part of the Sectorial programme ADER 2020, of the MADR, the
project 3.1.3. Development and modernization of the germoplasma collection in order
to preserve the long-term biological material national value.

REFERENCES
Anca Onache, Rădulescu I., Camelia Popa, 2008 – Clonal selection recently
homologate of tradional romanian varieties (Fetească regală, Fetească albă, Fetească
neagră). Analele Universităţii din Craiova, vol. XIII (XLIX), pp. 93-96.
Evans, R.G. 2000. The Art of Protecting Grapevines From Low Temperature
Injury.Proceedings A 50-a aniversare ASEV Annu. MTG., Seattle, Washington June,
19-23 iunie. pp. 60-72.
Sugar, D., R. aur, P. Lombard, și A. Gardea. 2003. Strategies for frost protection.
In: E.W. Hellman (Ed.) Oregon Viticulture. Oregon State University Press. Corvallis,
Oregon.
Țârdea C., Dejeu L., 1995, Viticultură, Editura didactică și pedagogică, București.
Vlădoianu Em., 1979 – Caracterizarea tehnologică a unor soiuri pentru vinuri albe
cultivate pe nisipurile irigate de la Dăbuleni, Dolj. Anale SCDCPN Dăbuleni, vol. III.
Editura Scrisul românesc, Craiova, pp. 150-155.
Vlădoianu Em., 1981 – Cercetări privind stabilirea tipului de vin ,,Roșior de
Oltenia”, pe nisipurile ameliorate din centrul viticol Dăbuleni. Lucrări ştiinţifice ale
SCCCPN Dăbuleni, vol. VI, pp. 217-224.

252
 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
 Food produce process ing
UNIVERSITY OF CRAIO VA
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

THE HOMEOPATHIC PRODUCTS USED IN PLANT PROTECTION:

AN ALTERNATIVE CHOICE

Rîndașu Ileana1, Ciceoi Roxana2, Stănică Florin1

1
University of Agronomic Sciences and Veterinary Medicine of Bucharest, Faculty of Horticulture,
2
University of Agronomic Sciences and Veterinary Medicine of Bucharest, Research Center for Study of
Food and Agricultural Products Quality, Laboratory of Diagnose for Plant Protection
*Correspondence author. E-mail: [email protected]

Key words: agrohomeopathy, disease, pest, trends

ABSTRACT
The paper presents the history and trends of the homeopathic products used in plant
protection. Also known as high-dilution or agrohomeopatic treatments, this method started with
the pioneering work of Kolisko in 1923, on wheat germination and Junker in 1928, on growth of
microorganisms. Their approaches can be mainly divided in two important types of studies:
effects on crop growth and effects on crop disease/phytopathology and pests control, both on
plant models and in field trials. An important contribution in the agrohomeopathic research is
brought by the isopathic model, where the same substance is used for the induction of a stress
and then, in a diluted form, for the cure. Our work synthetises the information gathered from
previous reviews made in 1984-2015 and other various papers published in the past years.

INTRODUCTION
The medicines taken for specific symptoms cause similar manifestations in
healthy organisms if taken a certain period of time (principle of similitude). The
opposite is also true: the medicines that cause symptoms in healthy organisms can
cure them when administered in sick individuals, in diluted, nontoxic doses.
In plants, the mechanism of action may imply metabolic changes which lead to
formation of secondary metabolic products related to the mechanism of defense of
plants (Lensi et al, 2010). Gangar has concluded that homeopathic medicines can
influence also the genetic activity in plants, this phenomenon being explained with the
help of experiments made on mungbean, gram, cotton, wheat etc.
The paper presents an analysis of the evolution of research made in
agrohomeopathy, from international publications, to put in evidence the level of
knowledge acquired between 1923 - 2017.

MATERIALS AND METHODS

For the present paper were used all the available online and offline
bibliographical references we could find, using the following international databases:
Web of Science - Core Collection (Journal Citation Reports, Derwent Innovations
Index, Thomson Reuters), SpringerLink Journals (Springer), Scopus (Elsevier),
ScienceDirect Freedom Collection (Elsevier), PROQUEST Central, HomBRex

253
(Homeopathy Basic Research experiments database). The period analysed in this
study was 1923-2017.

ISOPATHIC MODELS
The first studies on agrohomeopathy were performed by Kolisko on wheat
germination in 1923. Afterwards, in 1928, another researcher published a study about
the effects of high dilutions on microorganisms (Junker, 1928). These two researchers
were initiating a domain which lately was defined as agrohomeopathy, domain that
started to develop more intensely in the end of the 60's.
One of the earliest models belongs to Nitien et al, in 1969, with homeopathic
preparations of copper sulphate (Cuprum sulphuricum CH 15) for detoxification of
green pea previously intoxicated with the same substance in high dosage,
demonstrating a positive effect. Another study was performed in 2011 on wheat
seedlings pretreated with high concentrations of gibberellic acid expected to facilitate
the inhibitory effect of the same plant hormone in diluted doses, but the supposition
was not confirmed by the experimental results (Pfleger et al).
Another model consisted of seeds of Vigna unguiculata pretreated with
Natrium muriaticum and afterwards stressed with NaCl. In the pretreated group there
was an increase of seed germination compared with the control group stressed with
the same substance (Mondal, Sukul, 2012).
The action of Natrium muriaticum in dilutions 6 CH and 30 CH, in comparison
with NaCl solution of 5% was also studied on Phaseolus vulgaris. Natrium muriaticum
had a significative effect of increasing the vegetable growth, mainly in the 6CH dilution
(Lensi et al).
A comparative study was made on two plant models: Lemna gibba, an
autotrophic multicellular organism and Saccharomyces cerevisiae yeast, a
heterotrophic single-celled organism, stressed with arsenic and then treated with
decimal potency of Arsenicum album. Application of Arsenicum album on the Lemna
gibba bioassay yielded the largest effects compared to control groups, confirming the
hypothesis that complex organisms show stronger reactions to homeopathic
treatments (Jager, Scherr, 2011).

EFFECTS ON PESTS
The preparation made with Coccinella septempunctata from the adult beetle
gives the necessary protection against different types of pests, like Trioleurodes
vaporariorum (whitefly). Another efficient remedies against whitefly are made from the
tincture of a live parasitic wasp, Encarsia formosa (Kaviraj, 2012) or from Sulphur (CH
200), if the honeydew secretions leads to colonisation with sooty mould, turning the
leaves black (Maute, 2011).
An interesting study was made with the rosy apple aphid (Dysaphis
plantaginea Pass., considered a major pest in apple orchards) on apple seedlings, with
Lycopodyum clavatum CH 6, CH 15, CH 30 and a nosode of rosy apple aphid, in eight
independent, randomized and blinded experiments in growth chambers. The number of
juvenile offsprings was reduced after application of L.clavatum CH 15 and nosode CH
6 compared with the control group (Wyss and al).

EFFECTS ON DISEASES, GERMINABILITY AND CROP GROWTH

In most of the cases, the research of the effects on diseases was carried out
on artificially diseased organisms. (Table 1)

254
 Table 1
Summary of phytopathological bioassays (after Betti et al)

Authors Species/pathogen Treatment Working variable

Wild taro/ Disease symptoms,
Aggarval et al., Homeopathic
phytophthora fungal growth and
1993 treatments**
colocasiae spore germination
Cheema et al., Tomato/tobacco Clerodendrum Disease
1993 mosaic virus aculeatum, CH Thuja symptoms
Betti et al., Tobacco/tobacco Virus-induced
DH As2O3
2003 mosaic virus Hypersensitive lesions
Cheema et al., Papaya/papaya Homeopathic Disease
1986 mosaic virus treatments symptoms
Mulberry/ m. Plant growth, nematode
Datta, 2006 CH Cina
Incognita infection
Mango, guava,
Khanna and tomato/ Homeopathic
Post-harvest
Chandra, pestalotia spp., treatments
losses
1989 fusarium and adjuvants
roseum
Khanna and Spore respiration rate,
Chandra, Different fungi DH treatments organic acid pool in
1992 spores
Wheat, tomato/
Rivas et al., Seed and spore
Alternaria CH treatments***
1996 germination
solani
Apple/
Rolim et al., Powdery mildew
podosphaera CH treatments****
2001 symptoms
leucotricha
CH Thuja, nitric
Reed okra/
Saxena et al., acid, Sulphur, Fungal spore
seed-borne
1987 Calcarea carb., germination
fungi
Teucrium Q
Cowpea /
Sukul and Plant growth,
Meloidogyne CH Cina
Sukul, 1999 nematode infection
incognita
D, C = decimal, centesimal potency; H = Hahnemannian potency; ** = Kali iodatum, Arsenicum
album, Blatta orientalis, Thuja occidentalis; ***= Arsenicum album, Calcarea, Cuprum, Ferrum
metallicum, Lycopodium, Natrum, Phosphorus, Selenium, Silicea, Sulphur; **** = Kali iodatum,
Lachesis trigonocephalus, Staphysagria, Sulphur, Oidium lycopersici.

Khanna and Chandra, obtained good results in the control of rot in mango,
guava and tomatos, suppressing the spore germination and respiration of the fungi
Alternaria alternate, Colletotrichum gloeosporioides, Fusarium roseum and
Gloeosporium psidii.
Sinha and Singh obtained significative results in the control of fungi producing
aflatoxins in stored products, with Sulphur CH 200 which inhibited 100% the growth of
Aspergillus parasiticus. Other remedies (Silicea terra and Dulcamara) reduced the
growth of the fungi with 50% and the production of the toxin with 90%. Verma et al
evaluated the effect of homeopathic remedies Lachesis and Chimaphila both in CH
200, before and after the inoculation of the tobacco mosaic virus (TMV) and obtained a
reduction with 50% of virus content in tobacco leaf discs. In apple seedlings, two
sprays of Staphysagria 100 CH at intervals of 12 days reduced powdery mildews
255
caused by Podosphaera leucotricha (Rolim et al). Rolim used also Staphysagria, this
time in the 30 CH potency for tomato plants, in greenhouse and reduced the severity of
early blight, as well as with Phosphorus 30 CH and isotherapics of A. solani in 30 and
60 CH.
A three year project on the control of dark leaf spot caused by Alternaria
brassicicola in cauliflower made by Betti et al concluded that As2O3 DH 35 reduced
significantly the infection on cauliflower heads compared to control.
Other investigations are made on healthy plants for the determination of the
stimulation of the growth and on abiotically stressed plants (table 2).
The test organism most frequently used was wheat. The most commonly used
stressor was arsenic.
Table 2.

Overview of bioassays with abiotic stress (after Betti et al)

Authors Species Stress Treatment Work variable

Auquiere et CH Ethanol, Shoot growth, weight of
Wheat Ethanol Lysine
al, 1988 Lysine shoots
Novic et al., Shoot growth; fresh and dry
Wheat Dark germination DH Aurum
1990 weight of shoots
DH NaCl, DH Shoot growth; fresh and dry
Kovac et al., NaCl, CuCl,
Wheat CuCl, DH weight of shoots, grains and
1991 K2Cr2O7
K2Cr2O7 roots
DH Platinum,
Mercurius,
Cadmium, Shoot growth; fresh and dry
Lehner et
Wheat Dark germination Plumbum, weight of shoots, grains and
al., 1991
Cuprum, Aurum, roots
Argentum nit,
Sulfur
Shoot growth; fresh of
Lauppert, shoots; dry
Wheat Dark germination DH CuSO4
1995 weight of shoots, grains and
roots
Betti et al.,
Wheat As2O3 DH As2O3 Shoot growth
1997
Brizzi et al.,
Wheat As2O3 DH As2O3 Germination
2000
Binder et al.,
Wheat As2O3 DH As2O3 Shoot growth
2005
Brizzi et al.,
Wheat As2O3 DH As2O3 Shoot growth
2005
Carvalho et DH Arnica Shoot growth, parthenolide
Feverfew Adaptation
al., 2003 montana content
Carvalho et CH Natrum Shoot growth, chlorophyll
Water shortage
al., 2004 Feverfew muriaticum and proline content
Carvalho et CH Arnica Shoot growth, parthenolide
Feverfew Adaptation
al., 2005 montana content
Projetti et
Lentil CuSO4 CH CuSO4 Root growth
al., 1985
Steffen,
Yeast CuSO4 DH CuSO4 In vitro growth
1985
Tighe, 2005 Cress NaCl CH NaCl Shoot growth, germination
D, C = decimal, centesimal potency; H = Hahnemannian potency; K = Korsakovian potency;
256
 CONCLUSIONS
1. The difficulties of previous studies described in literature lies in the fact that
no complete medical material has been developed with specific indications of
phytopathology or pest control, although there are data reported by different authors.
The methodology varies from case to case, using various treatments, various
concentrations, and there is no standardization that allows identical treatments.
2. It is necessary to use new research models to properly assess the potential
of the domain. In particular, we have found that there are only a few studies on fruit
trees, most of the studies being made on wheat and vegetables.
3. There are only a few studies conducted in the field and no specific
methodologies have been established for different cultures. Most studies were
conducted in growth chambers and seedlings. New studies are needed to develop
methodologies for treatments applied directly to cultures, but the development of the
work strategy must first be done in laboratory conditions to control more easily the
environment.

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Models with plants, microorganisms and viruses for basic research in homeopathy.
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Betti L, Trebbi G, Majewsky V et al, 2009. Use of homeopathic preparations in
phytopathological models and in field trials: a critical review. Homeopathy; 98: 244-
266.
Jager T, Scherr C, Simon M, Heusser P, Wolf U, Heusser P, Baumgartner S,
2011. Investigation of Arsenic stressed yeast (Sacharomyces cerevisiae) as a
bioassay in homeopathic basic research. The Scientific World Journal; 11: 568-583.
Junker H, 1928. The effect of extreme dilutions on microorganisms. Phluger's
Archiv fur die Gesamte Physiologie; 219: 647-672.
Khanna KK, Chandra S, 1989. Further investigations of the control of storage rot
of mango, guava and tomato fruits with homeopathic drugs. Indian
phytopathology;3:436-440.
Khanna KK, Chandra S, 1992. effect of homeopathic drugs on respiration of
germinating fungal spores. Indian phytopathology; 45: 348-353.
Lensi MM, Siqueira TJ, Silva GH, 2010. A pilot study of the influence of Natrum
muriaticum 6cH and 30cH in a standardized culture of Phaseolus vulgaris L. Int J High
Dilution Res [cited YYYY Month dd]; 9(30): 43-50. Available from:
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Kaviraj V, 2012. Homeopathy for farm and garden, Narayana Verlag, Kandern;
148-149, 161-162, 167-169.
Kolisko L, 1923. Physiologischer und physicalischer Nachweis der Wirksamkeit
kleinster Entitaten, Verlag der Kommende Tag AG, Stuttgart.
Mondal S, Sukul NC, Sukul S, 2012. Natrum mur 200c promotes seed germination
and increases total protein, chlorophyll, rubisco and sugar in early seedlings of cowpea
under salt stress. Int J High Dilution Res;11:128.
Maute C, 2014. Homeopathy for plants. Narayana Verlag, Kandern; 50.
Nitien G, Boiron J, Marin A, 1969. Acao de doses infinitesimais de sulfato de
cobre sobre plantas previamente intoxicadas por essa substancia; acao da 15
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Homeopatica Brasileira, Rio de Janeiro.

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 Pfleger A, Hofacker J, Scherer- Pongratz W, Lothaller H, Endler PC, 2011. The
effect of extremely diluted agitated gibberellic acid (10c-30) on wheat seedling
development - a two researcher pilot study. Complement Ther Med; 19:164- 169.
Rolim PRR, Brignani NF, Silva JM, 2001. Controle de oidio da macieira por
preaparacoes homeopaticas. Fitopatologia brasileira; 26:436.
Sinha KK, Singh P, 1983. Homeopathic drugs - inhibitors of growth and aflatoxin
production by Aspergillus parasiticus. Indian phytopathology; 36: 356-357.
Verma HN, Verma GS, Verma VK, Krishna R, Srivastava KM, 1989. Homeopathic
and pharmacopeal drugs as inhibitors of tobacco mosaic virus. Indian phytopathology;
22:188-193.
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258
 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
 Food produce process ing
UNIVERSITY OF CRAIO VA
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

CONSIDERATIONS CONCERNING THE INFLUENCE OF VACUUM AND

PRESSURE CYCLIC PROCESS UTILISED TO IMPROVE THE
TENDERNESS OF ROMANIAN TRADITIONAL GOAT MEAT PRODUCTS
1x 2
Roşca Adrian , Roşca Daniela
1)
Faculty of Horticulture, University of Craiova
2)
Faculty of Electrical Engineering, University of Craiova
* Correspondence author. E-mail: [email protected]

Key words: compression / piercing, vacuuming, pressuring, Warner - Bratzler method

ABSTRACT
The paper presents a new method to increase the tenderness’ quality of the traditional
Romanian product “Goat Pastrama”. In order to decrease the wet curing / marinating period of
the goat meat, and in the same time to increase the tenderness’ quality of the product, the new
tenderizing process consists in one simultaneous compression and piercing step (2000 N), one
intensive cyclic vacuum process (0…- 0,85 bar), followed by certain steps of intensive cyclic
pressuring (0…9 bar) process. The paper presents characteristic shear force amounts obtained
by using Werner - Bratzler testing method for this type of cured - hot smoked final product
tenderized due to piercing / compression step, intensive cyclic vacuum process, followed by intensive
cyclic pressuring steps process, in comparison with no tenderized cured - hot smoked final
product.

INTRODUCTION
The word pastramă is etymologically rooted from the Latin pastor who means
shepherd, and from the Romanian a păstra which means "to keep" or "to preserve".
But the word is maybe more ancient and come so because Pastramă was shepherd‟s
meat food of lamb, sheep, goat or mutton. Pastramă was originally created as a way to
preserve meat before modern refrigeration. Traditionally Pas-trama is made from the
brisket (which comes from the lower chest of the steer), or from the navel (a small piece
cut from the muscle known as the plate), that are the tenderless parts of the animal‟s
carcass. For any meat Pastrama traditional making, the raw meat is brined, partly dried,
seasoned with various herbs and spices, then medium or hot smoked.
In industrial process Pastrama making consists in meat curing realized by fast
injection of brine usually containing preservation additives, or otherwise, in home-made or
small enterprise processing, marinated for long time in concentrated salt solution
(http://en.wikipedia.org/wiki/Pastrami).
In small / middle enterprise, after the brining process the meat is tenderized for 4
- 8 hours in massaging vacuum equipment (maximum relative vacuum - 0,65 …- 0,75
bar). For the same reason, in homemade or small enterprise process, the meat is
tenderized for 1 - 3 weeks in high concentration brine containing additional flavors
added. Then, the brined meat is cold smoked, and finally dried for long time in 2 - 8°C
ventilated air (Roşca & Roşca 2014, Roşca et al. 2014).

259
 Tenderizing is a process that breaks down collagens in meat to make it more
palatable for consumption.
There are several ways to tenderize meat: mechanical tenderization, such as
piercing; the tenderization that occurs through cooking, such as braising; ten-derizers
in the form of naturally occurring enzymes, which can be added to food before cooking
(examples of enzymes used for tenderizing: marinating the meat with vinegar, wine,
lemon juice or yogurt; brining the meat in a high concentration salt solution. Mechanical
tenderization actions produce multiple cuts in the meat muscle in order to increase the
surface area and thereby facilitate extraction and solubilization during the massaging
phase. Softening of the muscle is also obtained, making the meat more adaptable to the
cooking moulds (Tyszkiewicz & Klossowska 1996). Tenderization, pre-massage and
massage are closely inter-related, and not all products require the same mechanical
action. Thus the mechanical action must be intensified and adapted in order to
compensate for some of the negative cones-quences that may result in the product‟s
quality. This will depend on the rest of the process and, above all, on the presentation
and final quality of the product itself. In low-injection products where meat content
represents more than 80% of the final composition, meat quality is a determining factor
in mastication, while in more highly injected products, this is not as important as the
process and technology used (McGee 2004, Maddock 2008, Shewfelt 2000).
In order to reduce the time tenderizing process as much is possible, and to
improve the final product tenderness‟, this paper presents a new tenderizing method
based on compression / piercing step, one intensive cyclic vacuum process, followed by
intensive cyclic pressuring steps process.

MATERIAL AND METHODS

Material, equipment and processing method
This paper presents experimental research concerning a new method to obtain
a cured - smoked goat boneless brisket “Pastrama de capra” product, similar with
traditional Romanian assortment made by SC AVI-GIIS SRL Mihaesti, Valcea.
In order to produce “Pastrama de capra”, goat boneless brisket (4 pieces; 0,5
kg/piece; aprox. dimensions: length x width x height = 120 x 60 x 30 mm) was used
(according Animals Slaughter Certificate: 26 months, individual small farm).
In principle, at SC AVI-GIIS SRL Mihaesti, the traditional processing techno-logy
of this cured - smoked product type consists in: wet curing phase of entire pieces of the
meat in 12% curing salt concentration, during 2 - 3 weeks; drying/ripening phase in cold
ventilation (4 - 6°C) for 4 hours; middle smoking phase (45 - 50°C) for 2 days, followed by
a cold ventilation long sequence (4 - 6°C) for 8 hours (Roşca & Roşca 2014).
According to Romanian legislation, sodium nitrite (NaNO2), potassium or sodium
nitrate (KNO3 / NaNO3), or any chemical alternative curing substances are not
permitted in traditional cured - cooked types products usual processing, and no brine
injection is allowed (Roşca & Roşca 2014).
In order to determine the efficiency of the new tenderizing and curing method
proposed in this paper, one of these four pieces was used to make “Pastrama de
capra” in similar technological condition alike “SC AVI-GIIS SRL Mihaesti” is made.
The other 3 pieces were used to obtain “Goat Pastrama” made using the
proposed method.
To reduce the wet curing phase, and in the same time to improve as much is
possible the final product tenderness‟ quality, this paper propose a tenderizing method
based on certain steps: one simultaneous piercing and compression step, one
intensive cyclic vacuuming processing step, finally followed by intensive cyclic
pressuring process step.
260
 The meat‟s simultaneous piercing and compression processing is realized by
using Multi - needle piercing and compression device (designed and made by Un-
conventional Technologies and Equipment for Agro-Food Industry Laboratory - UTEFIL
within Faculty of Horticulture, in collaboration with Laboratory for Environmental
Protection in Industry - EPIL within Faculty of Electrical Engineering). This experi-mental
device consists in two parallel plates (W1.4571 stainless steel): the lower fixed plate
with 120 holes (ø5,5); the upper mobile plate with 120 needles (ø5, 200 conical sharp,
W1.4571) placed in the same shape and reciprocity distance as into the industrial brine
injection equipment (Figure 1) (Roşca & Roşca 2014).

Figure 1. Multi - needle piercing Figure 2. Experimental equipment

and compression device for cyclic vacuuming and
cyclic pressuring process

The meat‟s simultaneous piercing and compression is realized between the

two plates of the Multi - needle piercing and compression device that is actuated by the
crosshead of universal testing machines LBG 10 (within EPIL): maximum force 2000 N;
speed 100 mm/min. Due to the compression force the meat‟s initial height decreases
up to 20 mm final height.
For a better correlation of the research‟s results, all the four pieces (both one
piece to be processed at “SC AVI-GIIS SRL Mihaesti”, and the three pieces that will be
processed by using the method proposed in this paper, too) were simul-taneous
pierced and compressed (piece by piece), as above was presented.
In order to put in evidence the influence of cyclic vacuuming and cyclic pre-
ssuring process (CVCP) on meat tenderization, experimental equipment (EE - CVCP)
was used (Figure 2). In principle, EE - CVCP is composed in a vacuuming and pre-
ssuring process hydraulic cylinder (VP-HC) consisting in a cylindrical vessel (inner ø
80; length 180 mm) made in stainless steel W1.4571 and a piston made in food grade
Teflon. VP-HC is provided with a manometer gauge (0…12 bar; 1,6 class), and a
manovacuumeter gauge (-1 …1,5 bar; 2,5 precision class) when vacuuming process is
actuated. In order to evacuate the liquid / gas excess before and after VCPC, the
piston is provided with G1/4” tap connected to ø8 Rilsan tube.
EE - CVCP was designed and made by UTEFIL, in collaboration with EPIL.
(Roşca & Roşca 2014, UTEFIL Data Base 2005-2017).
In order to actuate the vacuuming and pressuring processes into EE - VCPC,
universal testing machine LBG 10 (within EPIL), was used.
After meat‟s simultaneous piercing and compression the new tenderizing
method presented in this paper proposes two main successive steps:

261
 - Cyclic vacuuming and de - vacuuming step of the pierced and compressed
meat‟s and a certain amount of brine (brine concentration: 1% salt solution; meat / brine
proportion: 2/1), too, into a processing vessel.
The cyclic vacuuming and de - vacuuming step lasts 15 min consisting in 5 consecutive
stages, each stage lasting 3 min: slow vacuuming up to - 0,85 bar (during 1 min), followed by
maintaining for 2 min at this vacuum level, followed by fast de-vacuuming up to the ambient
atmosphere (Figure 3).
During this step the blood is “squeezed” from the meat. At the end of this cyclic
step, the salt solution and the “squeezed” blood are replaced from the vessel, but the
meat remains into the vessel. Over the meat into the processing vessel a certain amount
of new brine is poured (brine concentration: 3% salt solution; meat / brine proportion: 1/1),
and the next processing step can begin.

Figure 3. Diagram for cyclic vacuuming

and de - vacuuming step

- Cyclic pressuring and de - pressuring step in the processing vessel of the

pierced and compressed meat and the brine at certain pressure amounts. During the
pressuring process, the pressure level is 4 - 5 times higher than during brine in-jection
in industrial equipment, and 3 - 4 times than the dynamic pressing / dropping during the
industrial massaging process. Therefore this large amount of pressure realizes uniform
tenderizing both meat‟s internal tissues nearby each piercing / per-foration, and the
external meat‟s internal tissues, too.
Each cyclic pressuring and de - pressuring step lasts 15 min consisting in 5
consecutive stages, each lasting 3 min: slow pressuring (during 2 min) up to 9 bar,
followed by maintaining for 1 min at this pressure level, followed by fast de-pressuring up
to the ambient atmosphere (Figure 4).
In this paper were considered certain successive repetitions lasing 60 min (CP
60), 90 min (CP 90) and 120 min (CP 120).
All the four meat pieces (1 pieces no tenderization - TRAD, brined in 12 % salt
concentration, during 2 weeks; each of 3 pieces CP 60, CP 90 and CP 120 (tenderized
as presented above) were ripened in cold ventilation (4 - 6°C) for 4 hours; middle smoked
(45 - 50°C) for 36 hours, followed by a cold ventilation long sequence (4 - 6°C) for 8 hours.

262
 Figure 4. Diagram for cyclic pressuring
and de - pressuring step

Tenderness evaluation using Warner - Bratzler method

Instrumental - objective methods are based on fundamental tests measure
properties that are familiar to mechanical engineers (strength, Poisson‟s ratio, Young‟s and
shear modulus). Mechanical tests cover a wide range of simple and rapid tests, including
puncture, compression, extrusion, shear, and others, which measure one or more textural
properties and are commonly used in quality control applications. Actual instruments tend
to be more and more sensitive to small differences between samples and are able to
detect trends in quality loss before they can be detected by humans. The most relevant
and utilized destructive texture and tenderness tests are puncture/ penetration test,
compression test, and Warner - Bratzler shear test.
In Warner - Bratzler test a blade cuts through a specimen, therefore the shear
force behavior gives information about tenderness, as well as the bite characteristic
products. The jig consists of a rigid frame supporting a shear bar. Interchangeable shear
blades fit into the frame. There are wide known two blade types: the square cut blade that
is mainly used for rectangular specimens and the notched (triangular or “V” blade) for
round cross-section specimens. The jig acts in direct compression for slicing / shearing
tests on products. Because of the good reproducibility of the results, this test is widely
used in food texture and tenderness analyzes (Xargayó et al. 2015, Xianzhong &
Shaofang 2011).
To perform interdisciplinary researches concerning general texture and
tenderness analysis, universal testing machines Lloyd Instruments LRXPlus 5 (0,5
accuracy class for force and extension) within UTEFIL is used since several years ago to
perform comparative texture measurements for fruits and vegetable firmness‟s (skin
strength, ripeness‟ degree), and for the tenderness of row-meat and cooked-meat. The
mechanical characteristics of the specimens as a function of force - deformation are
recorded and displayed by using specialized NEXYGENPlus control and measure-ment
software (www.lloyd-instruments.co.uk).
Due to collaboration between UTEFIL and EPIL an experimental equipment
Warner - Bratzler was made: special rigid frame (made in food-grade Teflon) sup-porting
a shear bar that permits interchangeable Warner - Bratzler shear blades (V and square
plate cut blade made in stainless steel DIN W1.4571) to slide fit into the frame (Figure 5)

263
(Roşca & Roşca 2011, Roşca & Roşca 2013, UTEFIL Data Base 2005-2017). During this
experiment, 100 mm/min cutting speed was used.
Goat Pastrami obtained using goat brisket tenderized CP 90 method, during
Warner - Bratzler testing shear force, is presented in Figure 6.

Figure 5. Universal testing machine Figure 6. Goat Pastrama made using CP 90

Lloyd Instruments LRXPlus 5 method, during Warner - Bratzler
and Warner - Bratzler shear force test
experimental equipment

RESULTS AND DISCUSSIONS

In order to determine the influence of tenderizing process on the final pro-duct
tenderness, Goat Pastrama pieces made by using traditional homemade method, or
tenderized by using the presented processing method, respectively, were sliced
according Warner - Bratzler shear force test; during these shear force tests, each of all
4 pieces of Goat Pastrama were sliced in 6 parts. Representative Warner - Bratzler
shear force diagram is presented in Figure 7.
In Table 1 are presented the maximum shear force amount and the shear
force average for each of the four of Goat Pastrama types, and in the same time,
presents the percentage average decrease of shear force‟s (in comparison with tra-
ditional homemade Goat Pastrama‟s tenderness TRAD) by using each processing
method, that demonstrate the tenderness quality increase of the product.
Table 1
Warner - Bratzler shear force for Goat Pastrama types
Sample Maximum shear force Shear force Decrease of shear
code min…max amount, N average, N force average, %
TRAD 281,78…323,55 301,46 -
CP 60 171,22…213,34 188,59 38,62
CP 90 148,11…173,82 159,11 47,25
CP 120 132,82…151,63 141,75 53,14

264
 Load (N)

150

100

50
Load at Maximum Load: 152.58 N
Machine Extension at Maximum Load: 26.67 mm
Stress at Maximum Load: 0.92474 MPa
Extension at Maximum Load: 26.67 mm
Work to Maximum Load: 2859.50 Nmm
Tensile Strength: 0.92474 MPa
0

0 10 20 30 40
Extension (mm)

Figure 7. Warner - Bratzler shear force diagram for Goat Pastrama made using
simultaneous piercing and compression, one cyclic vacuuming and de - vacuuming
step, finally followed by cyclic pressuring and de - pressuring step CP 90 method

CONCLUSIONS
Due to both high vacuum level and fast vacuuming and de-vacuuming step, and
high pressure level of the cyclic pressuring and de - pressuring steps, too, the pro-posed
processing method represents a much more intensively tenderizing method, than home-
made tenderizing method.
The main conclusion concerning the influence of simultaneous piercing and
compression, one cyclic vacuuming and de - vacuuming step, finally followed by cyclic
pressuring and de - pressuring step could be drawn from Table 1: in comparison with
traditional homemade Goat Pastrama‟s tenderness, each type of the proposed method
deter-mines an important tenderness‟ increasing (36,62…53,14%) of the this product.
The proposed method determines the shortest salt marinating / tenderizing period,
only 90-120 min, in comparison with 4-8 hours in massaging vacuum equipment in industrial
processing, or 2-3 weeks marinating time in homemade or small enterprise processing.
This paper opens further experimental researches focused on the influence of
similar cyclic vacuuming / pressuring process (higher vacuum or pressure level, shorter or
longer vacuuming and de-vacuuming steps, and pressuring and de - pressuring steps,
respectively) for making traditional products made in other much more tenderless parts of
animal‟s carcass.

REFERENCES
McGee H., 2004, On food and cooking, the science and lore of the kitchen. Scribner,
155-158, ISBN 978-0-684-80001-1.
Maddock R., 2008, Mechanical Tenderization of Beef, Research RKM & Knowledge
Management, pp. 77-98.
Roşca A., Roşca D., 2011, Instrumental texture evaluation - An objective measuring
method for quality assurance in food industry. Annals of the University of Craiova,
Biology, Horticulture, Food produce processing technology, Environmental engineering
Series, vol. XLI (LII), Universitaria Publishing House Craiova, 361-367.

265
 Roşca A., Roşca D., 2013, Instrumental texture evaluation - An objective method to
evaluate fresh vegetables quality. Journal Progress of Cryogenics and Isotopes
Separation, ICSI-ICIT Rm. Valcea, Vol. 16, issue 1, 54-60.
Roşca Daniela, Roşca A., 2014, The influence of pressuring and vacuuming cyclic
process on tenderizing Beef Pastrami, Analele Universităţii din Craiova, seria Agricultură
- Montanologie - Cadastru (Annals of the University of Craiova - Agriculture, Montanology,
Cadastre Series), Scientific Conference with International participation „Sustainable
Development in Agriculture and Horticulture” and Durable Agriculture - Agriculture of
future”, The Tenth Edition, Vol. XLIV/1/2014, 245 - 254, ISSN 1841-8317, http://anale.agro-
craiova.ro/index.php.
Shewfelt R. L., 2000, Consumer - friendly specifications to meet the demands of a
global market. Food Australia, 52, 311-314.
Tyszkiewicz I., Klossowska B.M., 1996, Mechanical Tenderization of Pork Meat:
Protein and Water Release due to Tissue Damage. Journal of the Science of Food and
Agriculture, Vol. 73(2), 179-185.
Xargayó M., Lagares J., Fernández E., Borrell D., Sanz D., 2015, The impact of
tenderization on increased slicing yield. http://en.metalquimia.com/upload/article.
Xianzhong Xu, Shaofang Yuan, 2011, An examination of the force generated from
incisor penetration into foods with different textural properties part I: Expe-rimental
observation, Journal of Texture Studies, Special Issue, Vol. 42, Issue 3, pp. 228-235.
Data Base. Unconventional Technologies and Equipment for Agro-Food Industry
Laboratory, 2005 - 2017.
www.lloyd-instruments.co.uk
http://en.wikipedia.org/wiki/Pastrami

266
 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
UNIVERSITY OF CR AIO VA  Food produce process ing
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

MATHEMATICAL MODEL FOR COMPRESSION STRENGTH

OF PORK SHOULDER MEAT TENDERIZED
BY USING MECHANICAL PROCESSES
1 2x
Roşca Daniela , Roşca Adrian
1)
University of Craiova, Faculty of Electrical Engineering
2)
University of Craiova, Faculty of Horticulture
* Correspondence author. E-mail: [email protected]

Key words: mathematical model, compression strength, Warner - Bratzler method

ABSTRACT
The paper presents a mathematical model for compression strength for shoulder pork
meat tenderized by using mechanical process. The mathematical model is based on
compression characteristic diagrams that are experimentally determined after pork shoulder
meat tenderi-zation by using statically pressing. In order to decrease the wet curing / marinating
period of the pork shoulder meat, the new tenderizing process consists in one statically pressing
step, one simultaneous compression and piercing step, followed by certain steps of intensive
cyclic vacuum processing. The paper presents characteristic shear force amounts obtained by
using Warner - Bratzler testing method for a cured - hot smoked final product tenderized by
pressing, piercing and cyclic vacuum processing, in comparison with no tenderized cured - hot
smoked final product.

INTRODUCTION
Tenderization is the mechanical action of producing multiple cuts in the meat
muscle in order to increase the surface area and thereby facilitate extraction and
solubilization during the massaging phase. Tenderization, pre-massage and massage are
closely interrelated, and not all products require the same mechanical action. Thus the
mechanical action must be intensified and adapted in order to compensate for some of
the negative consequences that may result in the product‟s quality. This will depend on
the rest of the process and, above all, on the presentation and final quality of the product
itself. Tenderizing process breaks down collagens in meat to make it more palatable for
consumption. There are several ways to tenderize meat: mechanical tenderization, such
as piercing; the tenderization that occurs through cooking, such as braising; tenderizers
in the form of naturally occurring enzymes, which can be added into food before cooking
(examples of enzymes used for tenderizing: marinating the meat with vinegar, wine,
lemon juice, yogurt (Institute of Food Technologists 1981, Maddock 2008, Tyszkiewicz et
al. 1996).
Cohesion of the muscles takes place thanks to the myofibrillar proteins which
have been extracted during the manufacturing process and which are found on the
surface of the muscle. These proteins form the exudates and, due to their gelling
capacity, act as glue between the muscles. It has been widely demonstrated in the
pertinent literature that the greater number of proteins extracted, the greater the stability
between muscles and therefore the better the sliceability and masti-cation. Extraction of
267
myofibrillar proteins is achieved through both mechanical and chemical actions. The
degree to which the muscle structure is opened will determine the final quantity of
proteins present in the exudates. This structure‟s openings is done by means of
tenderization, pre-massage and massage (Maddock 2008).
By means of certain additives, aside from the above-mentioned salt and
phosphates, such as carragenan and vegetable gums, muscle texture can be slightly
hardened and / or “plastified”, how-ever this alone will not be sufficient to compen-sate
for the meat‟s lack of firmness. It has been observed that the mechanical action of
tenderization does have a positive effect on this type of meat, because the texture is less
fragile due to an increased surface of contact between muscles (Maddock 2008,
Tyszkiewicz et al. 1996).
There are slight differences in the processing technology of cured - cooked
products, mainly depending on the size of the meat parts used for product making.
Curing brine is administered in all products, usually done by brine injection. Even
distribution of the injected brine is achieved by treating the injected meat pieces in a
meat tumbler, and when no tumbler is available, “resting periods” for the meat pieces are
needed. For some raw - cured products smaller amounts of curing brine are injected
directly into the muscle tissue to accelerate the curing process. This fast curing technique
significantly shortens curing periods, as curing substances migrate in both directions,
from outside to inside and from central to less central parts. But because of this
accelerated process, the curing flavor remains less intensive and texture of these
products remains softer then in products applying dry or wet curing. The shelf life is also
reduced significantly and most products are kept refrigerated. Typical products of this
fast-cured type are cured - smoked pork, beef, sheep and goat meat. Fast curing with
injection of curing brine will therefore remain the method of choice for rapid turn - over
cured - cooked meat products only (Maddock 2008, Tyszkiewicz et al. 1996).
During wet-salting process, the curing salt solution infiltrates the meat tissue and
at the same time liquid from the meat tissue is extracted by the salt surrounding the
meat. Depending on the size of the meat cuts, the curing process alone can last up to
several weeks for equal penetration of the meat cuts with curing salt (at temperatures of
about +4°C, a pork shoulder takes about two weeks, a leg of pork about four weeks)
(Tyszkiewicz et al. 1996, Xargayó et al. 2011) .
As exception to the common technology of using curing salt (containing nitrite or
nitrate, or a mixture of both), some well - known traditional cured - raw ham products
(e.g.“Parma Ham” and “Jamon Serrano” in Spain; “Jambon Savoie” in France; Romanian
traditional products “Ceafa Perperlita”), are made without nitrite, using common salt only.
Although no nitrite is used, a stable red color is achieved in these cured - raw ham
products. This red color derives from the natural meat color intensified by the drying and
ripening process (Roşca & Cirdei 2015, Tyszkiewicz et al. 1996, Xargayó et al. 2011). .

MATERIAL AND METHODS

Material, processing method and equipment
This paper presents experimental and theoretical research concerning a new
method to obtain a cured - hot smoked boneless shoulder “Ceafa Hituita” product similar
with traditional Romanian assortment “Ceafa Perpelita” made by SC AVI-GIIS SRL
Mihaesti, Valcea County.
In principle, the processing technology of this traditional cured - hot smoked -
cooked product type consists in: wet curing phase of entire pieces of muscle meat in 12 -
15% curing salt concentration, during 2 - 3 weeks; drying / ripening phase in cold
ventilation for 6 - 8 hours; cold smoke phase (20 - 25°C) for 2 - 3 days, followed by a short

268
sequence of hot smoke phase (80 - 85°C), for 4 - 6 hours (Roşca & Cîrdei 2015, Simion
et al. 2014).
According to Romanian legislation, sodium nitrite (NaNO2), potassium/sodium
nitrate (NaNO3 / KNO3), or any alternative curing substances are not permitted in tradi-
tional cured - cooked types products usual processing, and no brine injection is allowed
(Roşca & Cîrdei 2015, Simion et al. 2014).
In order to produce traditional cured - hot smoked - cooked product “Ceafa
Hituita”, 2 large pieces (long pieces from symmetrical both sides) of boneless pork
shoulder were used (Animal Slaughter Certificate: 18 months, 95…100 kg carcass; large
farm). Each of this large piece of boneless pork shoulder was cut in 2 equal small pieces
(aprox. dimensions: length x width x height = 150 x 70 x 45 mm).
To determine the efficiency of the new tenderizing and curing method pro-posed
in this paper, one of these small pieces was used to make “Ceafa Hituita” in similar
technological condition alike “Ceafa Perpelita” is made by SC AVI-GIIS SRL Mihaesti. The
other 3 small pieces were used to obtain “Ceafa Hituita” made by using the proposed
method.
In order to improve the final product tenderness, and in the same time to reduce
as much is possible the wet curing phase, this paper propose a tenderizing method based
on meat one pressing / compression step, one simultaneous compre-ssion and piercing
step, followed by intensive cyclic vacuum process.
The pressing / compression step is realized by introducing the meat between two
Teflon plates; the compression force is realized by actuating the crosshead of universal
testing machines LBG 10 (Environmental Industrial Engineering Laboratory - EIL, within
Faculty of Electrical Engineering), with maximum force 3500 N and speed 200 mm/min.
Due to the compression force the meat‟s initial height decreases up to 20…25 mm final
height. During the compression step, the testing machine‟s software plotted compression
force - deformation diagram.
The meat‟s simultaneous compression and piercing processing is realized by
using the experimental Multi - needle compression and piercing device (Figure 1). The
experimental equipment consists in two parallel plates (W1.4571 stainless steel): the
lower fixed plate with 120 holes (ø5,5); the upper mobile plate with 120 needles (ø5, 200
conical sharp, W1.4571) disposed in the same shape and reciprocity distance as into the
industrial brine injection equipment (Roşca et al. 2014, UTEFIL Data Base 2005 - 2017).
The meat‟s simultaneous compression and piercing is realized between the two plates of
the Multi - needle compression and piercing device that is actuated by the crosshead of
universal testing machines LBG 10 with maximum force 2000 N and constant speed 200
mm/min.
In actual massaging vacuum processing equipment (tumbler) the vacuum level
do not exceed - 0,7 bar. Recent American and European research papers con-cerning in
meat tenderizing recommend increasing the vacuum level up to - 0,95 bar (Maddock
2008, Xargayó et al. 2011).
This paper proposes the simultaneous curing and massaging of the tenderized
meat by using intensive cyclic vacuum processing. In principle, the experimental equip-
ment for Intensive Cyclic Vacuum Processing (ICVP) consists in vacuum pump and
vacuum processing vessel (Figure 2). The main characteristics of the vacuum pump
(HYVAC type): maximum flow rate up to 40 l/min; absolute pressure up to 50 mbar.
The vacuum processing vessel (stainless steel W1.4571; welding coef. 1)
permits vacuum experiments for absolute pressure up to 0,5 mtorr. A rotative mixing
device (mounted into the vacuum; stainless steel W1.4571) can be put in operation for
rotational motion (1 - 30 rot / min) by a special electromechanical transmission speed

269
variator. To observe the inlet vessel during vacuum process, one of the flanges is made
in transparent visor (high resistance polycarbonate) (UTEFIL Data Base 2005 - 2017).

Figure 1. Multi - needle compression Figure 2. Experimental equipment for ICVP

and piercing device
The experimental equipment for ICVP and Multi - needle compression and
piercing device, too, were designed and made by Unconventional Technologies and
Equipment for Food Industry Laboratory - UTEFIL within Faculty of Agriculture and
Horticulture, in collaboration with EIL.
The intensive cyclic vacuum process (ICVP) consists in the following steps:
- The compressed and pierced meat (each piece tenderized after pressing /
compression step at 3500 N, followed by simultaneous compression and piercing as
above was presented) is introduced into the rotative mixing device of the vacuum pro-
cesssing vessel that contain only 3% salted concentration brine (proportion 2:1 for brine
and meat, respectively); 10 rot / min rotational speed was set.
- Each intensive vacuum cycle lasts 40 min consists in 20 consecutive steps,
each step lasting 2 min: slow vacuuming up to - 0,85 bar (during 1 min), followed by
maintaining for 1 min at this vacuum level, followed by fast de-vacuuming up to the
ambient atmosphere (Figure 3).
- After finishing each 40 min intensive cyclic vacuum process, the used brine was
replaced by a fresh cold one. The used brine replace lasts 3…4 min, and during this
period the meat is relaxed after it was “squeezed” by the vacuuming process.
This paper presents the results obtained by using 3 types of intensive vacuum
cycles:
- 40 min intensive vacuum cycle (consisting in 20 consecutive steps, each
step lasting 2 min): ICVP 3500-40;
- 80 min intensive vacuum cycle (consisting in 40 consecutive steps, each
step lasting 2 min): ICVP 3500-80;
- 120 min intensive vacuum cycle (consisting in 60 consecutive steps, each
step lasting 2 min): ICVP 3500-120.
All the four meat pieces (1 pieces no tenderization - TRAD, brined in 12 % salt
concentration, during 10 days; each of 3 pieces tenderized by compression, simul-
taneous compressed and pierced at 3500 N, followed by intensive cyclic vacuum process
during 40 min, 80 min, 120 min, respectively) were ripened in cold ventilation for 6 hours,
then cold smoked 22 - 25°C / 6 hours and hot smoked 80 - 85°C / 4 hours.
Tenderness evaluation by using Warner - Bratzler method
The most relevant and utilized methods to estimate meat‟s tenderness are
compression test, and Warner - Bratzler shear test. During Warner - Bratzler test the
shear blade acts simultaneously compression and slicing / shearing of the product
(Wheeler et al. 2001, Xargayó et al. 2011, Xianzhong & Shaofang 2011).

270
 Figure 3. Intensive cyclic vacuum process diagram

To perform inter-disciplinary researches concerning tenderness analysis,

universal testing machine Lloyd Instruments LRXPlus 5 (within UTEFIL) was used.
Due to collaboration between UTEFIL and EIL, a Warner - Bratzler experi-mental
equipment was made. In principle, this equipment consists in a special rigid frame that
permits operative / fast fit and sliding of interchangeable Warner - Bratzler shear blades
(UTEFIL Data Base 2005 - 2017). The experimental research performed for this paper,
100 mm/min cutting speed was set.
Mathematical model for compression strength to tenderize the meat
The compression strength behavior is represented by the internal stress within the
meat when certain compression force determines the decrease of the sample height‟s
amount (deformation during compression force action) (Graiver et al. 2009).
In this paper, a mathematical method based on MathCAD software was pro-
posed to study the compression strength‟ behavior.
Therefore, to determine the deformation‟s variation diagrams when a certain force
is applied, experimental data were used to describe the compression force matrix (Roşca
et al. 2014):
 F15 F14 F13 F12 F1 
 5 
 F2 F24 F23 F22 F2 
 
M :  F35 F34 F33 F32 F3 
 F5 F44 F43 F42 F4 
 4 
 F5 F54 F53 F52 F5 
 5 ,
(1)
and the vectors that define the each deformation determined by each compression force

271
  d1 
 d2 
v :   d3 
 d4 
d 
 5 , (2)
where each d := k, k + m...l, (k :=0; m := 0,5; l := 10).
After several preliminary analytical calculi, it was observed that the nearest
function which approximates the experimental force - deformation diagram, is the fifth
degree polynomial function defined by the relation (Roşca et al. 2014)
F( d )    d 5    d 4    d 3    d 2    d (3)
where α,..., ε represent influence coefficients that have to be determined solving the
relation (Roşca et al. 2014)
soln := lsolve (M, v) (4)
The mechanical work when compression force is applied, has to be obtained
solving the equation (Roşca et al. 2014)
d2

W mec   F d   d d  (5)
d1

RESULTS AND DISCUSSIONS

In order to determine the influence of compression process on meat‟s com-
pression strength and the final product tenderness‟ too, the meat was stressed with 3500
N compression force; the experimental diagram is presented in Figure 4a. Using rel. (1),
rel. (2) and rel. (4), for 3500 N compression force, were determined
 0,1735 0,1734 0,1733 0,1732 0,173    1,472 
   
 2,1755 2,175 4
2,175 2,1752
2,175   0,033 
M :   4,8775 4,877 4
4,877 3
4,8772

4,877  sol n   0,417 
  
 7,3215 7,3214 7,3213 7,3212 7,321    5,212
 5   68.40 
 9,752 9,7524 9,7523 9,7522 9,752   
;
and after solving rel. (3), it was obtained the particular polynomial function that best
describes the diagram‟s evolution of meat‟s deformation when compression force is
applied
F (d )  1,472  d 5  0,033 d 4  0,417  d 3  5,212  d 2  68,40  d
(6)
The force - deformation diagrams plotted by using rel. (6) for 3500 N, is pre-
sented in Figure 4b. The comparison between diagrams in Figure 4a and Figure 4b
when 3500 N compression force is applied, confirms the correctness of the proposed
mathematical model.
Table 1 presents the experimental mechanical work and the theoretical me-
chanical work obtained by using the mathematical model, when compression force 3500
N is applied.
The differences between the mechanical work determined by using experi-
mental method and the mathematical model are smaller than 7,8%, that confirms the
correctness of the proposed mathematical model. These small differences could be
explained by viscoelastical deformation of the tissues‟ structure when meat is stressed
during compression force acts.

272
 Table 1
Mechanical work for 3500 N compression force

Method Mechanical work, Nmm

Experimental 23840
Mathematical Model 21990

Load (N)

Load at Maximum Load: 3555.11 N

Stress at Maximum Load: 0.13466 MPa
Extension at Maximum Load: 21.46 mm
Work to Maximum Load: 26823.89 Nmm
3000

2000

1000

000

0.0 5.0 10.0 15.0 20.0 25.0

Extension (mm)

4000

3600

3200

2800

2400

F ( d) 2000

1600

1200

800

400

0
0 2 4 6 8 10 12 14 16 18 20
d
b
Figure 4. Compression diagram for 3500 N
a - experimental; b - mathematical model
273
 To evaluate the efficiency of the 3 types of the proposed method (ICVP 3500-40;
ICVP 3500-80; ICVP 3500-120) on the tenderness of cured - hot smoked “Ceafa Hituita”
final product, the Warner - Bratzler shear force experimentally deter-mined for these
types processing method was compared with the shear force amounts obtained for
“Ceafa Hituita” made using traditional method (TRAD).
In Table 2 are presented the maximum shear force amount and the shear force
average for each “Ceafa Hituita” type. Table 2 presents too, the decrease of percentage
average of shear force, in comparison with traditional homemade “Ceafa Hituita”s
tenderness, that demonstrates the final product tenderness‟ increasing.
Table 2 presents the influence of compression / pressing process, followed by
simultaneous compression and piercing, and finally followed by intensive cyclic vacuum
process on the tenderness of cured - hot smoked on “Ceafa Hituita” final tenderness‟: the
final product tenderness‟ improving with 35,04% for ICVP 3500-40, with 41,76% for ICVP
3500-80 and with 47,18% for ICVP 3500-120, respectively.
Table 2
Warner - Bratzler shear force for “Ceafa Hituita”

Sample Maximum shear force Shear force Decrease of shear

code min…max amount, N average, N force average, %
TRAD 163,40...208,93 184,64 -
ICVP 3500-40 109,48...139,57 119,46 35,04
ICVP 3500-80 92,11...122,62 107,53 41,76
ICVP 3500-120 86,73...106,49 97,52 47,18

CONCLUSIONS
In order to improve the cured - hot smoked “Ceafa Hituita” tenderness‟, the
proposed tenderizing method consists in compression / pressing process, followed by
simultaneous compression and piercing, and finally followed by intensive cyclic vacuum
process.
All these mechanical tenderization steps causes pressing and relaxation of the
muscle structure and breaking up of the cells, making the membranes more permeable
and increasing mobilization of the proteins up toward the surface of the muscle when the
intensive cyclic vacuum process is applied. The degree to which the muscle structure is
opened will determine the final quantity of proteins present in the exudates.
Due to much intensive osmosis phenomena when simultaneous compre-ssion
and piercing followed by cyclic vacuum process is applied, meat„s compression strength
decreasing deter-mines faster brine‟s infusion into the meat‟ tissues.
The salt percentage in all the final product “Ceafa Hituita” made by using any
one of the three types of the proposed method (ICVP 3500-40; ICVP 3500-80; ICVP
3500-120) was 2,6...3% (according to the analyses performed by Sanitary and Veterinary
Direction, Dolj).
As one of the most recommended analyze method, the Warner - Bratzler shear
force test offered objective results concerning the influence of compression / pressing
process, followed by simultaneous compression and piercing, and finally followed by
intensive cyclic vacuum process on final products tenderness‟.
In main, the method described in this paper has two important advantages:
- Reducing the wet curing phase from 2 weeks to only 120 min (for the best
results).

274
 - Improving the tenderness of Romanian traditional cured - hot smoked pro-duct
“Ceafa Hituita” (35,04...47,18%), in comparison with the traditional product “Ceafa
Perperlita” made with no tenderized meat.
- Low salt content (up to 3 %) and uniform distribution in the product volume.
Each mechanical tenderizing step determines significant changes of specific
strain within the meat‟s tissues, which determines the tenderness‟ improvement.
Considering only physically issues, these mechanical tenderizing steps reduce the
meat‟s mechanical characteristics amounts of the final product.
The main conclusion drawn in this paper refers the correctness of the mathe-
matical model, proved both by the diagrams‟ configuration and by the intermediary or
maximum amounts similarities‟ between the numerical tenderizing diagrams, and the
experimentally diagrams.
Further general and specific conclusions could be draw after this mathe-matical
model will be applied for other types of meat, before and after the same or other
mechanical tenderizing methods. The data presented in this paper can be important for
all the specialists interested in decreasing the wet curing period and in tenderness
improvement too, of the traditional meat products.

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Graiver N., Pinotti A., Clifano A., Zaritzky N., 2009, Mathematical modeling of the
uptake of curing salts in pork meat, Journal of Food Engineering, vol. 95/4, 533-540.
Institute of Food Technologists, 1981, Sensory evaluation guide for testing food and
beverage products, Food Technology, 35 (11), 50-59.
Maddock R., 2008, Mechanical Tenderization of Beef, Research RKM & Know-ledge
Management, pp. 77-98.
Roşca A., Cîrdei P., 2015,Mathematical model for pork boneless neck tenderizing to
produce Romanian traditional product “Ceafă Perpelită”, The 17-th International Conference
of Scientific Paper, Scientific Research and Education in the Air Force - AFASES, Brasov,
ISSN 2247-3173, 41-50. http://www.afahc.ro/ro/afases/2015/afases.
Roşca Daniela, Csatlos C., Roşca A., 2014, Considerations on compression
strength of tenderized pork meat by using mechanical processing, Analele Universităţii din
Craiova, seria Agricultură - Montanologie - Cadastru (Annals of the University of Craiova -
Agriculture, Montanology, Cadastre Series), Scientific Conference with Inter-national
participation „Sustainable Development in Agriculture and Horticulture” and Durable
Agriculture - Agriculture of future”, The Tenth Edition, Vol. XLIV/1/2014, 234 - 244, ISSN
1841-8317. http://anale.agro-craiova.ro/index.php.
Simion A., Cârdei R. P., Roşca A., Bădescu M., Roşca, Daniela, 2014, Mathe-matical
model based on Warner - Bratzler analyze method concerning tenderized pork boneless
loin in order to produce Romanian traditional product “Cotlet Perpelit“ type, International
Symposium ISB - INMA - TEH‟ 2014, National Institute of Research - Development for
Machines and Installations Designed to Agriculture and Food Industry - INMA, Bucharest,
71-84.
Tyszkiewicz I., Klossowska B.M., 1996, Mechanical Tenderization of Pork Meat:
Protein and Water Release due to Tissue Damage, Journal of the Science of Food and
Agriculture, Vol. 73(2), 179 - 185.
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protocol, USDA - ARS U.S. Meat Animal Research Center,http://www.ars.usda.gov
Xargayó M., Lagares J., Fernández E., Borrell D., Sanz, D., 2011, The impact of
tenderization on increased slicing yield, 73-97. http://en.metalquimia.com/upload.

275
 Xianzhong Xu, Shaofang Yuan, An examination of the force generated from incisor
penetration into foods with different textural properties part I: Experimental observation,
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Laboratory, 2005 - 2017.

276
 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
 Food produce process ing
UNIVERSITY OF CRAIO VA
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

DETERMINING THE HEALTH STATUS OF

PLATANUS X ACERIFOLIA TREES BY MEANS OF TOMOGRAPHY

Sima Sanda (Petredeanu)1*), Baciu Aurelian Adrian 1)

Faculty of Horticulture, University of Craiova, 13 A.I. Cuza Street, 200585, Craiova, Romania
*) corresponding author, e-mail: [email protected]

Key words: regeneration, London Plan Tree, trunk health,

ABSTRACT
Urban green areas, faced with increasing levels of real estate and social pressure, are
suffering a decrease in quality and value. This fact draws our attention to elements that play the
role of memory benchmarks, landscape values, which can define and characterize the evolution
of a society. Centennial trees or trees of an esthetic, historical, memorial, and social value are
also cultural landmarks that require a great deal of attention considering their value, correlated
with a lack of vision in terms of protecting and preserving their historical significance and
importance for the landscape. In the current context, the public or private green areas of
Bucharest host a number of 110 protected trees, declared monuments of nature -- according to
the List of protected trees, maintained by the Romanian Academy.

INTRODUCTION
The decrease in the number and size of green area accentuates
environmental risks present in the urban environment, and have an immediate
negative impact on the quality of life and general health of the population. This is why it
is necessary to create new green areas, especially in the most crowded urban centers,
or, where they already exist, to manage them properly (Chiriac et al.,2009).
Law 24/2007 on the regulation and administration of green areas in urban
environments stipulates that "the State recognizes every person's right to a healthy
environment, free recreational access to public green areas, the right to contribute to
the setup and care of green areas, to the creation of tree and bush lines, with the
observance of the current provisions of the law." The same law defines urban green
areas as a mosaic-like network or a system of semi-natural ecosystems, characterized
by vegetation (woods, trees, bushes, flowers, herbs), which include parks, squares,
planted street linings or clear, unproductive plots within the built area.
The national program for improving the quality of the environment by setting up
green areas in municipalities allowed the funding of projects for the creation of new
parks, or for the rehabilitation and expansion of existing parks (Toti, 2008).
The locations and plots assigned to green areas in European municipalities
depend on current management as well as the existing traditions in various cities.
Considering the pressure that green areas are exposed to, their size as well as the
changes they face become an important indicator in the assessment of policies
planning the "urban green." But, in addition to ensuring an optimal area of greenery,
European municipalities must also consider the requirements of various groups of
users, the demands for the variety of forms of recreation, the improvement of the urban
environment.
277
 The most important are parks, because of their area, facilities and functions
they meet (Stănescu, 2008).
A green area of this type in Bucharest includes an arboreal vegetation
comprised of sycamores, maple trees, poplars, carob trees, chestnut trees, linden
trees and firs. Because the area is populated by interesting specimens of centennial
sycamores, belonging to the Platanus x acerifolia species, it is known as "Sycamore
Park" (Parcul cu Platani). The land underwent several transformations, and it currently
belongs to the National Bank of Romania.
The study herein aims to analyze the specimens of Platanus x acerifolia
existent in the Sycamore Park from the point of view of their heritage value, in order to
perform a complex analysis of the characteristics of each specimen and determine its
health status, with the end goal of preserving and protecting this heritage.
All the on-site determinations and analyses will inform the decision-making
process regarding measures of regeneration and preservation of highly valuable
specimens at the Sycamore Park.

MATERIAL AND METHODS

The study was carried out on a pool of 47 Platanus x acerifolia specimens, this
species lending its name to the place - Sycamore Park - with 31 of them being
centennial and having various health statuses. These 31 specimens are featured on
the list of protected trees, managed by the Romanian Academy.
Each specimen was analyzed individually, according to a scheme created as
per the principles of Sustainable management, having an individual analysis sheet
assigned, containing aspects regarding the morphology of the tree (basic anatomic
elements: trunk, crown), the internal structure of the trunk (assessed in accordance
with the tomographic analyses carried out), the overall health and appearance of the
tree.
Before the trees were analyzed, they were coded, taking into account the
acronym of the Latin name of the genus and species (Pa – Platanus x acerifolia) as
well as the tree identification number (01, 02 etc.).
Tomographic profiles were carried out with the help of the ARBOTOM system,
containing highly sensitive sensors for measuring the duration of transmission of the
operator-generated impulses, so as to obtain a high-resolution diagram of the
transversal arrangement of the areas affected by wood decomposition.
The number of sensors installed was established according to trunk shape and
diameter. Sensor 1 was always placed towards north, so that the affected areas within
the trunk could be established later. The nails used for anchoring sensors are inserted
into the wood (the ritidom/bark must be fully pierced), and in order to intercept the
shape of the trunk as faithfully as possible in its transversal section, the nails must be
anchored in points of minimum and maximum radius to the tree axis (fig. 1). In order to
obtain 2D diagrams, safety nails are installed on the same plane. The average height
at which the tomographic profiles were carried out is approximately 20cm. In the case
of visibly damaged specimens, two tomography sessions were carried out: one at the
basis of the trunk, and one at a higher level, in order to ascertain whether the
degradation is advanced or in remission.

278
 Fig. 1. Location of sensors (user's handbook)

According to the degree of degradation, the analyzed sycamores were

classified in four categories:
- group a – healthy (0% degradation);
- group b – slight damage (1-15 % degradation);
- group c – medium damage (16-30 % degradation);
- group d – advanced damage (> 30 % degradation) (fig. 2).

a – healthy b – slight damage

c – medium damage d – advanced damage

Fig. 2 – Categories of degradation of the analyzed trees

RESULTS AND DISCUSSION

The analysis of the existing relations between the monitored features is of
great importance, as it allows the selection of genotypes that present an association of
several useful features. The value of the correlation coefficient may vary between ± 1.
Values close to +1 indicate a strong positive correlation; those close to -1 indicate a
strong negative correlation, whereas values close to 0 indicate the fact that there is no
correlation between the two features.

279
 Fig.3. The correlation between trunk diameter and age, for the sycamores at the
Sycamore Park - NBR.

The correlation between trunk diameter (cm) and calculated age (years) shows
that there is a close dependency between the two, with a highly significant correlation
coefficient (r = 0.9995); between the two determinations there is a direct positive
relation (fig. 3).
This proves the fact that the more a tree ages, the higher the quantity of
biomass it accumulates, including the one stored in the trunk.
Yet if we study the correlation between the circumference of the trunk and its
degree of degradation, we find that this correlation is positive, but with a value of r =
0.32599, meaning the degree of degradation is less influenced by trunk size (fig. 4).
A value r = 0.326874 of the correlation between the degradation of sycamore
trunks and their calculated age indicates the fact that, in addition to age, there are
several other elements influencing their health status (wounds -- cavities, periodic
pruning methods etc).

Fig.4. The correlation between trunk degradation and trunk circumference, for the
sycamores at the Sycamore Park - NBR.

280
 Fig.5. The correlation between trunk degradation and age, for the sycamores at the
Sycamore Park - NBR.
Out of the total 47 specimens of sycamores analyzed, it was concluded that 8
trees (17.02%) are healthy, 18 trees (38.30%) are slightly damaged, 10 trees (21.28%)
suffer medium degradation, while 11 sycamores (23.40%) suffer from advanced
degradation of the trunk (table 1).
In public parks and gardens, in the green areas of boulevards and avenues as
well as in green areas managed by the Public Administration, tree cutting (shearing,
pruning) techniques are carried out by qualified staff, in unity and harmony with the
other elements of the existing landscape architecture.
This study was carried out as it is compulsory to perform a periodical (5-10
years) identification of the physiological status of trees for the establishment and
enforcement of the required works - pruning, cutting down, maintenance and
regeneration - in order to render the administration and protection of the city's
dendrologic fund more efficient.

CONCLUSIONS
1. For determining the physiological state of trees in order to establish and perform
maintenance works it is recommended to perform health check-ups every 5-10 years.
2. Trees' health status can be determined by using the ARBOTOM system, which
carries out tomographic profiling.
3. According to the degree of damage, established by tomography, the sycamores
were grouped into four categories: healthy, slight damage, medium damage and
advanced damage.
4. By analyzing the correlations between various parameters, we find various
degrees of correlation.
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Cais în V (2005). Creşterea arborilor pentru 25 de ani şi clima. Mediul ambiant
2(20): 17-21.
Dumitru C, Hună C, Stanciu M (2009). Spațiile verzi – o problemă a urbanizării
globale. Calitatea vieții XX, 3-4:249-270.
Fritts H C (1990). Modeling tree-ring and environmental relationships for
dendrochronological analysis. Modeling of Forest Growth Responses to Environmental
Stress. 368-382.
281
 Toti M (2008). Dezvoltarea durabilă în contextul lansării oficiale a strategiei
naţionale. Infomediu Europa 7.
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Table 1.
Summary of sycamores analyzed by tomography

No. Degree of Trunk Calculated age

Crt Trunk diameter
Tree code tomography degradation circumference years
No (cm)
sessions (%) (cm) (Ø x 6,956)
1 Pa 01 2 50/40 330 105 151
2 Pa 02 1 40 395 126 181
3 Pa 03 1 31 316 101 145
4 Pa 04 2 51/35 404 129 185
5 Pa 05 1 0 312 99 142
6 Pa 17 1 35 312 99 142
7 Pa 18 1 35 267 85 122
8 Pa 19 1 0 288 92 132
9 Pa 20 1 5 253 81 116
10 Pa 21 2 29/23 218 69 99
11 Pa 22 1 16 278 88 127
12 Pa 23 1 16 245 78 112
13 Pa 24 1 0 305 97 139
14 Pa 28 2 23/45 262 83 119
15 Pa 30 2 37/48 320 102 147
16 Pa 43 1 20 210 67 96
17 Pa 44 1 25 355 113 162
18 Pa 45 1 22 311 99 142
19 Pa 46 1 0 291 93 134
20 Pa 47 1 11 363 116 167
21 Pa 48 1 8 341 109 157
22 Pa 50 2 9/14 316 101 145
23 Pa 51 1 3 286 91 131
24 Pa 52 1 22 276 88 127
25 Pa 53 1 16 270 86 124
26 Pa 54 2 14/0 293 93 134
27 Pa 55 1 1 182 58 83
28 Pa 56 1 11 264 84 121
29 Pa 57 1 0 243 77 111
30 Pa 59 1 28 248 79 114
31 Pa 60 1 12 264 84 121
32 Pa 61 1 23 267 85 127
33 Pa 62 1 3 237 75 108
34 Pa 63 1 11 350 111 160
35 Pa 64 1 6 264 84 121
36 Pa 65 1 23 399 127 183
37 Pa 66 1 7 258 82 118
38 Pa 67 1 2 273 87 125
39 Pa 68 1 0 240 76 109
40 Pa 69 2 54/36 393 125 180
41 Pa 73 1 13 332 106 152
42 Pa 74 1 1 372 118 170
43 Pa 75 2 35/17 342 109 157
44 Pa 76 2 19/6 255 81 116
45 Pa 77 1 0 310 99 142
46 Pa 126 1 0 335 107 154
47 Pa 127 1 12 378 120 173

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 Food produce process ing
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Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

BRIEF AGROBIOLOGICAL AND TECHNOLOGICAL CHARACTERISTICS

OF NEW TABLE GRAPES CANDIDATE-VARIETIES

Simeonov Iliyan1*, Ivanov Miroslav1, Nakov Zdravko1, Iliev Anatoly1

* Institute of Viticulture and Enology, Pleven, Bulgaria, e-mail: [email protected]

Key words: vine, table grapes candidate-varieties, agrobiological and technological

characteristics

ABSTRACT
An ampelographic study on the newly-selected Vit, Nayden and Miro table grapes
candidate varieties at the Insitute of Viticulture and Enology, Pleven was carried out. The study
found that, according to the time of the consumer's maturity of the grapes, the Vit and Miro
varieties belong to the group of early maturing, while Nayden to the group of medium-ripening
varieties of vines. The vines of newly selected table grapes candidate varieties are characterized
by medium to strong growth, good habits, high actual fertility and productivity, which are
relatively constant in years. By mechanical composition of the grapes, the candidate varieties
studied are typically dessert, with specific characteristics of the grapes and grains, suitable for
fresh consumption. Candidate varieties Vit and Nayden obtained by intra-hybridization are
characterized by high sensitivity to low winter temperatures. They can be grown in all wine-
growing regions of the country where the risk of freezing of vines is less. The candidate variety
Miro, obtained through the interspecific hybridization, has a genetically determined increased
resistance to low winter temperatures, mana and oidium and can be grown in all wine-growing
regions of the country. As regards of the appearance and organoleptic profile of the grapes, it
closely matches to the best table grapes varieties of Vitis vinifera L.

INTRODUCTION
As a result of mankind development, the selection of vine varieties from their
natural habitats and their propagation and distribution, a large number of varieties have
been grown nowadays, as their main set is specific for each country. These varieties
differ in their morphological signs, agrobiological features and technological qualities,
and their grapes could be used for various purposes (Ivanov, 2011). The soil and
weather conditions in Bulgaria are extremely favourable for planting and growing
vineyards with table grapes varieties. That is one of the reasons for the growing
interest of the viticultural science in the country to expand the available gene pool of
table grapes varieties through comprehensive selection-genetic research. Grapes
quality and the products derived from it has been directly dependent on the variety as a
means of production with its specific ampelographic features and the complex of
factors of the environment and the production conditions by which the soil and climatic
resources of nature had been used in the most rational way (Katerov et al., 2005). The
combination of the natural conditions and the potential of the varieties was a
prerequisite for grapes production of individual quality, superior appearance and
excellent organoleptic qualities (Todorov, 1998).

283
 Recent studies have shown that, on the background of the constantly changing
market situation of grapes and wine, the increasing agrotechnical level and the
dynamical climate changes, the variety structure in Bulgaria has not been flexible
enough with prevailing varieties whose grapes ripen from mid-August to mid-
September (Bachvarova et al., 2007, Krumov, 2014). That revealed the need of
creating and introducing new vine varieties with valuable biological and economic
qualities, increasing the share of early and late ripening seeded and seedless varieties
with big berries. The experience of the global viticulture science has demonstrated that
even today, gender hybridization has been the most current and efficient selection
method for obtaining new diverse grapevine varieties (Valchev, 1978, 1990; Stoev,
1984; Alleweldt & Possingham 1988). As a result of the achievements in the field of
vine genetics and selection, a number of new valuable table grapes and wine varieties
have been created in Bulgaria in the recent decades (Lazarov et al., 2004).
The objective of this study was to make a brief economic characteristic of the
newly-selected at IVE – Pleven table grapes candidate-varieties.

MATERIAL AND METHODS

The candidate-varieties Vit, Nayden (intra-species varieties) and Miro
(interspecific variety) has been object of the study. The ampelographic investigation
was carried out during the period 2014-2016 in vineyard (hybrid section) at the
Experimental base of IVE-Pleven. The vines were grown on improved Guyot training
system, at planting distance 2.20/1.30 m, grafted to Berlandieri x Riparia SO4
rootstock. For the ampelographic investigation of the variety the methodology
described in Bulgarian Ampelography volume 1 (Katerov et al., 1990) was used. It
included the following indicators:
- Botanical description of a cluster and a berry;
- Agrobiological characteristics – Phenological observations – budding,
flowering, interspersing, consumption maturity; Actual fertility – developed eyes ratio,
fruit shoots ratio, number of clusters, actual fertility rate; average yield per vine, kg.
- Mechanical analysis of a cluster and a berry – mass and size of the cluster
and the berry, ratio of rachis, seeds, skin, mesocarp, theoretical yield, tearing and
pressure;
- Technological characteristics – Chemical composition – sugars content (%),
and titratable acids (g/dm 3).

RESULTS AND DISCUSSIONS

Candidate-variety V 6-18 (Vit)
Candidate-variety V 6-18 (Vit) was obtained by intra-species hybridization from
the crossing of Dunav x Kondarev 10 varieties at the Experimental base of the Institute
of Viticulture and Enology - Pleven by M. Ivanov, Z. Nakov and I. Simeonov.
Botanical description
Cluster. The cluster is large (26.7/16.5 cm), conical, loose to semi-compact.
The stem is medium long, medium thick. The rachis is green. The average mass per
cluster is 785.5 g (Fig. 1).
Berry. The berry is large (28.46/18.96 mm), egg-shaped. The skin is dark blue,
thin and fragile, with wax coating. The texture is fleshy-juicy. The taste is harmonious,
neutral. The average mass per 100 berries is 633.3 g.
Agrobiological characteristics
Candidate- variety V 6-18 (Vit) is an early ripening red seedless table grapes
variety. Its grapes ripen at the end of August. The vegetation period duration is about
150 days. The vines of candidate-variety Vit have medium to intense growth, good
284
habitus and abundantly covered with foliage. At ground and stem training systems the
candidate-variety has good fertility and yield, relatively constant per years. Vit is not
liable to putting forth catkins and milerandage and it is distinguished for very good
pollination and fertilization. The candidate-variety Vit is obtained by intra-species
hybridization and it is susceptible to low winter temperatures, powdery mildew, downy
mildew and gray rot. At ground and stem cultivation it could be applied improved Guyot
training with cane length of 14-16 eyes. For grapes quality improvement green pruning
treatments should be done – suckering and removal of part of the leaves around the
clusters.

Figura 1. Candidate-variety V 6-18 (Vit)

Technological characteristics
According to the grapes mechanical composition the candidate-variety is a
typical tables grape variety. The berries ratio in the cluster is 97.50%, of the rachis –
2.50%. The skins in the berries are 3.41% and mesocarp – 96.59. At consumption
maturity the grapes contain 15-17% sugars and 5-6 g/dm3 titratable acidity. The grapes
are suitable for fresh consumption and meeting the market demand for quality
seedless table grapes during the second half of August.
Candidate-variety V 8-2 (Nayden)
Candidate-variety V 8-2 (Nayden) was obtained by intra-species hybridization
from the crossing of Dunav x Rusalka 3 varieties at the Experimental base of the
Institute of Viticulture and Enology - Pleven by M. Ivanov, Z. Nakov and I. Simeonov.
Botanical description
Cluster. The cluster is large (19.2/17.2 cm), conical, loose to semi-compact.
The stem is medium long, medium thick. The rachis is green. The average mass per
cluster is 817.7 g (Fig. 2).

285
 Figura 2. Candidate-variety V 8-2 (Nayden)

Berry. The berry is large to very large (38.69/18.10 mm), drop-like (specific).
The skin is deep purple, thin and delicate. The texture is fleshy, crisp, and the taste –
neutral, harmonious. The berry stem is medium, with middle bed. The rachis is medium
long, pale green. The average mass per 100 berries-873.3 g.
Agrobiological characteristics
Candidate-variety V 8-2 (Nayden) is medium ripening red seedless table
grapes variety. In the Pleven region its grapes ripen early in September. The
vegetation period duration is about 170 days. The vines of candidate-variety Nayden
have medium to intense growth, good habitus and plenty of foliage. At ground and
stem training systems the candidate-variety is characterized with good fertility and
yield, constant per years. Nayden is not liable to putting forth catkins and milerandage
and it is distinguished for very good pollination and fertilization. The candidate-variety
is susceptible to low winter temperatures and downy mildew but it is relatively resistant
to powdery mildew and gray rot. V 8-2 (Nayden) might be grown at ground and stem
improved Guyot training system with cane length of 14 to 16 eyes. For grapes quality
improvement green pruning treatments should be done during the vegetation –
suckering and removal of part of the leaves around the clusters.
Technological characteristics
According to the grapes mechanical composition candidate-variety Nayden is a
typical tables grape variety. The berries ratio in the cluster is 97.06%, of the rachis –
2.94%. The skins in the berries are 4.08%, the seeds 1.38%, and mesocarp – 94.54%.
3
At consumption maturity its grapes contain 14-16% sugars and 5.5-6.5 g/dm titratable
acidity. The grapes of the candidate-variety are suitable for fresh consumption and
storage. It has very good transportability. After storage it keeps its appearance and
taste qualities.
Candidate-variety V 1-40 (Miro)
Candidate-variety V 1-40 (Miro) was obtained by interspecific hybridization
from the crossing of Plevenski Favorit x Nadezhda varieties at the Experimental base

286
of the Institute of Viticulture and Enology - Pleven by M. Ivanov, Z. Nakov and I.
Simeonov.
Botanical description
Cluster. The cluster is very large (23.0/16.5 cm), cylindrical-conical, semi-
compact. The stem is medium long, thick, woody at the base. The rachis is green. The
berry stem is medium long, thick with average large cone-like bed. The average mass
per cluster is 862.8 g (Fig. 3).
Berry. The berry is very large (32.07/23.15 mm), oval. The skin is greenish-
yellow in colour, thin and delicate, with wax coating. Mesocarp texture is fleshy-juicy.
The taste is harmonious and neutral. The average mass per 100 berries is 955.0 g.
Agrobiological characteristics
Candidate-variety V 1-40 (Miro) is early ripening white table grapes variety. Its
grapes ripen in late August. The vegetation period duration is about 150-160 days. The
vines of candidate-variety Miro have medium to intense growth and plenty of foliage. At
ground and stem training systems it is distinguished with very good fertility and yield,
constant per years. The candidate-variety Miro is not liable to putting forth catkins and
milerandage and it is characterized by very good pollination and fertilization. The
candidate-variety Miro has higher resistance (1-2 scores) to fungal diseases compared
to the control varieties belonging to Vitis vinifera L. It has high resistance to low winter
temperatures and downy mildew and very good restoring capacity after frost. At ground
and stem cultivation it could be applied improved Guyot training system with cane
length of 14 to 16 eyes. For grapes quality improvement green pruning treatments
should be done during the vegetation – suckering, pinching off and removal of part of
the leaves around the clusters.

Figura 3. Candidate-variety V 1-40 (Miro)

Technological characteristics
According to the grapes mechanical composition candidate-variety V 1-40
(Miro) is a typical tables grape variety. The berries ratio is 97.28%, of the rachis –
2.72%. The skins from the structure of the berries are 3.54%, the seeds 1.31%, and

287
mesocarp – 95.02%. At consumption maturity its grapes contain 15-16% sugars and
5.5-6.5 g/dm3 titratable acidity. The grapes of the candidate-variety Miro are suitable
for fresh consumption and storage. It has very good transportability. After storage it
keeps its appearance and taste qualities.

CONCLUSIONS
According to the term of grapes consumption maturity, the candidate-varieties
Vit and Miro belong to the group of early ripening while Nayden to the medium ripening
grapevine varieties.
The vines of the newly selected table grapes candidate-varieties are
characterized by medium to intense growth, good habits, high actual fertility and
productivity, relatively constant per years. In grapes mechanical composition the
candidate-varieties are typically table grapes, with specific characteristics of the cluster
and the berries, suitable for fresh consumption.
The candidate-varieties Vit and Nayden, obtained by intra-species
hybridization are characterized by high susceptibility to low winter temperatures. They
can be grown in all vine-growing regions of the country where the risk of frost is
smaller.
The candidate-variety Miro, obtained by interspecific hybridization, has a
genetically determined increased resistance to low winter temperatures, downy mildew
and powdery mildew and can be grown in all the vine-growing regions of the country.
Regarding the appearance and organoleptic profile the grapes closely rivals the best
table grapes varieties of Vitis vinifera L.

REFERENCES
Alleweldt G., Possingham J. V. 1988. Progress in graprvine breeding. TAG
Theoretical and Apllied Genetics 75(5), 669-673.
Bachvarova, S., Pandeliev S., Dimitrova V., Strelkova D., Georgiev Ts., Popov K.,
Kovatchev V., Vachevska D., Yaneva V. 2007. National program to encourage the
production of table grapes in the Republic of Bulgaria from 2006 to 2020. Edition of
MAF, January, 2007.
Ivanov, M. 2011. Results of Interspecies Hybridization of Table Grapes Varieties,
Dissertation thesis, Plovdiv, 181p.
Katerov К., еt al., 1990. Bulgarian Ampelographia, General ampelographia,
Publisher of BAS, Vol. І, 296 p.
Katerov K., Peykov V., Ivanov M. 2005. Practical viticulture with ampelography.
Dionysus, Sofia, 272 p.
Krumov S., 2014. Agrobiological study of table grape varieties in Kyustendil area,
Dissertation thesis, Kyustendil, 186 p.
Lazarov I., Kostova V., Kirillov B. 2004. Vine varieties Bulgarian selection, Sofia,
184 p.
Stoev К., 1984. Physiology of grapes and the basis of its cultivation. BAS, Sofia,
V. 3, 328 p.
Todorov I., 1998. Problems of viticulture in Bulgaria on the eve of the 21st century,
Viticulture and enology, 4, 35-36.
Valchev V., 1978. Selection-Genetic Studies in Creation through Hybridization of
New Wine Varieties Resistant to Mildew (Pl. Viticola) and Cold, Dissertation thesis,
Pleven, 183 p.
Valchev V., 1990. Intraspecific and interspecific hybridization of the vine.
Habilitation, Pleven, 272 p.

288
 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
 Food produce process ing
UNIVERSITY OF CRAIO VA
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

CULTIVAR INFLUENCE UPON LETTUCE PRODUCTION, CULTIVATED IN

AUTUMN IN AN ECOLOGICAL SYSTEM

Ştef Adrian Valeriu1, Apahidean Alexandru Silviu1, Cărbunar Mihai2, Bei Mariana2,
1 1
Apahidean Alexandru Ioan *, Domocoş Daniela
1
University of Agricultural Sciences and Veterinary Medicine, Faculty of Horticulture,
3-5 Mănăştur Street, Cluj-Napoca, 400372, Romania,
2
University of Oradea, Faculty of Environmental Protection,
B-dul Gen.Magheru 26,.Oradea, Romania
Corresponding author. E-mail: *[email protected]

Key words: garden lettuce, cultivar, planting age, production

ABSTRACT
Garden lettuce is grown for the head and leaves, which are mainly consumed in green
salads, simple or combined with other green vegetables. It is important in nutrition due to its high
content in vitamins and mineral salts. It is cultivated in the field, early spring and autumn, as well
as in greenhouses and polyethylene tunnels, to obtain production during winter-spring or
autumn-winter period. Being a species with a short period of vegetation is cultivated in the
system of successive crops, before or after a basic crop, as well as in associated crops. In
temperate climate, salad culture is not practiced during the summer because in long day
conditions associated with high temperatures, plants emit floral stems in most varieties.The
experience was carried out in 2016, in the western part of Romania, in Husasău de Tinca,
county of Bihor, where 14 salad varieties were grown in an ecological system.

INTRODUCTION
Garden lettuce is grown for leaves and heads that are used in fresh or
prepared form. It is demanded by consumers throughout the year, being cultivated
both in the field and protected areas in successive or associated crops.
Lettuce heads and leaves contain 4-8% dry substance, 2-3.5% carbohydrates,
1-1.6% protides, vitamins B1, B2, C (5-20 mg) P and E, carotene (1-3 mg) and mineral
salts: potassium-260 mg, iron 1.2-1.7 mg, calcium, phosphorus - 40 mg, magnesium -
24 mg, per 100 g of fresh product. Leaves also contain latex, which prints the bitter
taste (Gherghi et al., 2002). It is a lightly digestible, low caloric intake (16-20
calories/100 g), corresponding to a dietary regimen. Lettuce that gets through winter in
open field has a higher content in vitamins than in forced or spring crops (Apahidean &
Apahidean, 2016). Consumed fresh, lettuce contributes to the revitalization of muscle
tissue, brain and nerves, maintains the fluidity of the blood, is a good diuretic,
stimulates appetite, etc. (Soare & Duţă, 2008). Lettuce consumption reduces the risk of
heart disease, cancer and cataracts. It is rich in fibres that help reduce cholesterol
(Lagunovschi-Luchian & Vânătoru, 2016).
Lettuce because it has a high nitrate value, over 2500 mg NO 3/kg, is included
in the group of high nitrate vegetables. This level is influenced by both genetic and

289
ecotoxic factors, so measures can be taken to obtain productions with lower nitrate and
nitrite content (Soare & Duţă, 2008).
Lettuce is a long day plant, conditions in which it has a short period of
vegetation and forms floral stems before forming the head. Sensitivity to the length of
the day occurs between the end of May and the first half of June, during which the
length of the day is over 14 hours (Still, 2007). Plant growth can be influenced by
various physiological treatments (Okuda et al., 2017). Under short conditions, it forms
a rich foliage and large head, which do not pass into the floral stem phase (Ciofu et al.,
2003; Posta, 2008).
Optimal temperature for leaf growth and head formation is around 16oC, and
o
for the formation of floral stems and flowers, 20-22 C (Stan et al., 2003).
Lettuce production is influenced by the cultivar used, the culture period and the
culture system (Stoleru, 2013). At a protected crop during the spring, with different
cultivars, Szabo et al., 2010, obtained plants with an average weight of 353.0 to 474.0
g/plant. The average yield was 46.0 t/ha and the maximum yield of 54.77 t/ha was
achieved by the Limax cultivar.

MATERIALS AND METHODS

Experience was carried out in 2016, in the western part of Romania, in
Husasău de Tinca, county of Bihor where a lettuce culture was carried out in an
ecological system.
Village is located in the Miersig Plain, which consists of a higher stretch of flats
with a slight slope and terraces located below the Hidişelului (Taşadului) Hills and
another lower, in the west, close to Criş river channel. Northern and southern
boundaries are given by Crişului Repede and the Crişului Negru Meadow. South-
western boundaries are between Colector-Culiser and Crisul Negru, where a low-lying
area, extending at 90-105 m, which extends to the low plain of Crişul Negru, the
approximate limit passing south of the Tica- Tulca.
Biological material consisted of 14 salad varieties of several types,
recommended for the organic farming system:
- Roman Lettuce – represented by the following varieties: Dark Green, Blonde
Maraichere, Blonde Lente a Monter;
- Letuce type Batavia – represented by the following varieties: Long Stading
Batavian, Jester;
- Leaf lettuce – was represented by Lola Rosa and Lola Bionda;
- Head forming lettuce - Anueme, Gloire de Mantes, Merveille de 4 Saison,
Grass Blonde Peresuble, Laituie Silvesta and Laituie Appia;
Dark Green - roman salad that forms a lax head of light green to dark green.
Leaves are fleshy with a clear, crisp, and very savory main nervure.
Blond Maraichere - roman salad, which forms a tall, elongated but rather large
head. Outside leaves are light green.
Blonde lente a Monter - roman salad that forms a compact head. Leaves have
shape of a spatula, long petiolate pale green. It has a good headdress even in the
warm summer conditions.
Long Standing Batavian - Batavian salad with a bulky and high head. Leaves
are very thick in light green color. It is one of the most resistant batavian salads in the
issue of floral stems.
Jester - is a Batavian salad with a very beautiful look. It forms a large rosette of
very voluminous leaves. Leaves are large, half-embossed, toothed edges, bright green
pigmented with reddish spots. leafs are crisp and tasty.

290
 Anuenue - is a salad that has the ability to grow up in warm soil. Leaves are
bright green. It is adapted to high temperatures.
Kwiek - is a vigorous variety with green leaves with a slight reddish tinge.
Forms a medium-sized head.
Gloire De Nantes - forms a thick head with slightly light-colored leaves, light
green. It resists well at high temperatures.
Merveille of 4 Saisons - is a fast growing and vigorous variety. It forms an
elongated colored ruby head to light red. It can be cultivated in all seasons, but it
prefers more spring and summer.
Lollo Rossa - is a salad with brown green leaves and the red hot tip. It's very
tasty. It resists the heat.
Lollo Bionda - forms a light curly green leaf rosette.
Grosse Blonde Paresseuse - forms a large flattened head. Leaves are light
green and the interior leaves are pale green. It resists the heat.
Laituie Sylvesta- is a bright green rustic head salad. Leaves are crisp and very
tasty.
Laituie Appia - is a kind of salad of green head, with finely crispy, glossy, very
tasty leaves. Resistant to the issue of floral stems.
Culture was set up in autumn in two different epochs and in two different
culture systems (protected and unprotected). For the first epoch, planting took place on
12.09.2016, in the field, and the second epoch was set up in a polyethyelene tunnel,
planting the seedlings on 15.10.2016. Culture was established by seedling produced in
alveolar trays with 104 cells per tray. The soil mix for sowing was made of peat and
peat (with neutral pH). Planting was done 35 days after sowing. The planting distances
were 30 cm both between the rows and between the plants per row. 15 days after
planting a nettle macerate treatment was performed. Harvesting took place between
October and November, staggered according to variety and place of culture.

RESULTS AND DISCUSSIONS

Field crop production in autumn crops was between 2.70 kg/m 2 for Anuenue
variety and 3.73 kg/m2 for Dark Green variety (Table 1). Compared with experience
average, the varieties of Anuenue, Kwiek, Lollo Rossa, Lollo Bionda, Gloire de Nantes,
Merveille des 4 Saisons, Grosse Blonde Paresseuse and Laitue Silvesta produced
lower yields. Dark Green, Blonde Maraichère, Blonde Lente à Monter, Long Standing
Batavian, and Laitue Pomme Appia exceeded the average with 5.28-23.10%.
Dark Green variety produced a yield of 3.73 kg/m 2, the difference in production
compared to the control being very significant and in Blonde Lente à Monter the
production was 3.56 kg/m 2, with a distinctly significant production difference (Table 1).
Polyethylene tunnel crop production ranged from 3.23 kg/m 2 at Kwiek and 4.32
2
kg/m in Dark Green variety (Table 2). In comparison with the average of the
experience, the varieties Anuenue, Kwiek, Lollo Rossa, Lollo Bionda, Gloire de Nantes,
Merveille des 4 Saisons, Grosse Blonde Paresseuse and Laitue Silvesta produced
lower yields. Dark Green, Blonde Maraichère, Blonde Lente à Monter, Long Standing
Batavian, and Laitue Pomme Appia exceeded the average with 5.11-16.13%.
Dark Green variety obtained a production of 4.32 kg/m 2, the difference in
production compared to control being very significant, and at Blonde Lente à Monter
2
variety, the production was 4.25 kg/m , with a distinct difference in production (Table
2). A significant difference in production was also recorded in at Laitue Pomme Appia
variety.

291
 Table 1
Cultivar influence on the production of lettuce cultivated in the autumn in the field

No. Variant Average production Difference to Significance

kg/m2 % culture average of the
2
(kg/m ) difference
1 Dark Green 3.73 123.10 0.70 ***
2 Blonde Maraichère 3.42 112.87 0.39 *
3 Blonde Lente à Monter 3.56 117.49 0.53 **
4 Long Standing Batavian 3.19 105.28 0.16 -
5 Jester 2.90 95.70 -0.13 -
6 Anuenue 2.70 89.10 -0.33 0
7 Kwiek 2.73 90.09 -0.30 0
8 Lollo Rossa 2.75 90.75 -0.28 0
9 Lollo Bionda 2.77 91.42 -0.26 0
10 Gloire de Nantes 2.74 90.43 0.29 0
11 Merveille des 4 Saisons 2.81 92.74 0.22 -
Grosse Blonde 2.87 94.72 0.16 -
12
Paresseuse
13 Laitue Silvesta 2.90 95.70 0.13 -
14 Laitue Pomme Appia 3.40 112.21 0.37 *
Average 3.03 100.00 - -
DL P5% 0,25; DL P1% 0,47; DL P 0,1% 0,62

Table 2
Cultivar influence upon production of lettuce cultivated in autumn in polyethylene
tunnel

Variant Average production Difference to Significance of

kg/m2 % culture average the difference
(kg/m2)
Dark Green 4.32 116.13 0.60 ***
Blonde Maraichère 3.91 105.11 0.19 -
Blonde Lente à Monter 3.97 106.72 0.25 *
Long Standing Batavian 4.25 114.24 0.53 **
Jester 3.73 100.26 0.01 -
Anuenue 3.51 94.35 -0.21 -
Kwiek 3.23 86.82 -0.49 00
Lollo Rossa 3.58 96.73 -0.14 -
Lollo Bionda 3.45 92.74 -0.27 0
Gloire de Nantes 3.26 87.63 -0.46 00
Merveille des 4 Saisons 3.57 95.96 -0.15 -
Grosse Blonde Paresseuse 3.58 96.23 -0.14 -
Laitue Silvesta 3.65 98.11 -0.07 -
Laitue Pomme Appia 4.13 111.02 0.41 **
Average 3.72 100.00 - -
DL P5% 0,24; DL P1% 0,39; DL P 0,1% 0,55

Table 3
292
 Cultivar influence on the production of lettuce cultivated in autumn in open field and
polyethylene tunnel

Variant Average production Difference to % field production

Polyethylene Open culture compared to
tunnel Field average polyethyelene
(kg/m2) tunnel
Dark Green 4.32 3.73 0.59 86.34
Blonde Maraichère 3.91 3.42 0.49 87.46
Blonde Lente à Monter 3.97 3.56 0.41 89.67
Long Standing 4.25 3.19 1.06 75.05
Batavian
Jester 3.73 2.90 0.83 77.74
Anuenue 3.51 2.70 0.81 76.92
Kwiek 3.23 2.73 0.50 84.52
Lollo Rossa 3.58 2.75 0.83 76.81
Lollo Bionda 3.45 2.77 0.68 80.28
Gloire de Nantes 3.26 2.74 0.52 84.04
Merveille des 4 3.57 2.81 0.76 78.71
Saisons
Grosse Blonde 3.58 2.87 0.71 80.16
Paresseuse
Laitue Silvesta 3.65 2.90 0.75 79.45
Laitue Pomme Appia 4.13 3.40 0.73 82.32

Analyzing the behavior of the 14 cultivars in the two crop systems (protected
and unprotected), it was found that in the protected crop system, yields were higher
with 0.5-1.6 kg/m2 compared to the unprotected crop system (Table 3).
Dark Green, Blonde Maraichère, Blonde Lente à Monter, Kwiek, Lollo Bionda,
Gloire de Nantes, Grosse Blonde Paresseuse and Laitue Pomme Appia produced at
least 80% unprotected crop production compared to the protected system.

CONCLUSIONS
Based on the results obtained from the research on lettuce grown in open field
and in polyethylene tunnel, in the specific conditions in the western part of Romania, in
ecological system, using 14 cultivation, the following conclusions were drawn:
- crop production in the field, in autumn culture, was between 2.70 kg/m 2 for
the Anuenue variety and 3.73 kg/m2 for the Dark Green variety;
- production of autumn lettuce in polyethylene tunnel ranged from 3.23 kg/m2
2
at Kwiek and 4.32 kg/m in Dark Green variety;
- In protected crop system, yields were higher by 0.5-1.6 kg/m2 compared to
the unprotected crop system, depending on the cultivar;
- Dark Green, Blonde Maraichère, Blonde Lente à Monter, Kwiek, Lollo
Bionda, Gloire de Nantes, Grosse Blonde Paresseuse and Laitue Pomme Appia have
obtained a yield in unprotected crop system of at least 80% compared to the protected
culture system.

REFERENCES
Apahidean Al.S., Apahidean Al.I., 2016, Legumicultura, Ed. AcademicPres, Cluj-
Napoca, pp. 202-206.

293
 Ciofu R., Stan N., Popescu V., Chilom P., Apahidean S., Horgoş A., Berar V.,
Lauer K.F., Atanasiu N., 2003, Tratat de Legumicultură, Ed.Ceres, Bucureşti, PP.900-
913.
Draghici E., 2002, Legumicultura, Ed.Granada, Bucuresti, pp. 217-220.
Gherghi A., Burzo I., Bibicu M., Mărgineanu L., Bădulescu L., 2002, Biochimia şi
fiziologia legumelor şi fructelor, Ed. Academiei Române, Bucureşti, pp.180-185.
Lagunovschi-Luchian V., Vînătoru C., 2016, Legumicultură, Ed. ALPHA MDN,
Buzău pp.301-305.
Okuda N., Miya Y., Yanagy T., Yamaguchi K., 2017, Effects of End of Day Lighting
after Night Chilling Treatment on Growth and Development of Lettuce, Environ Control
Biol., 55 (1), pp. 7-11.
Poşta Gh., 2008, Legumicultură, Ed. Mirton, Timişoara, pp.215-220.
Soare R., Duţă A., 2008, Tehnologii legumicole alternative, Ed.Universitaria,
Craiova, pp. 230-235.
Stan N., Munteanu N., Stan T., 2003, Legumicultură Vol.III, Ed. Ion Ionescu de la
Brad, Iași, pp. 180-190.
Stoleru V., 2013, Managementul sistemelor legumicole ecologice, Ed. Ion Ionescu
de la Brad, Iași, pp. 230-234.
Szabo J. A., Apahidean Al. S., Laczi E., 2010, Study of some lettuce (Lactuca
sativa L.) cultivars in covered crop conditions, Revista Lucrări ştiinţifice, seria
Horticultura, Iaşi,pp. 201-204.
Still D.W., 2007, Genome mapping and molecular breeding in plants, Vegetables,
Ed.Springer-Verlag, Berlin, Vol.5 pp. 101-107.

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ANNALS OF T HE  Horticu lture
 Food produce process ing
UNIVERSITY OF CRAIO VA
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

ON VINEYARD CULTIVATION IN ECOLOGICAL SYSTEM IN DEALUL

BUJORULUI VINEYARD UNDER THE CLIMATIC CONDITIONS OF 2016

Tabaranu Gabriel1, Viorica Enache1, Donici Alina1

1*
Research Station and Development for Vineyard and Vinification Bujoru, România,
e-mail: [email protected]

Key words: grapevine, pathogens, vineyard

ABSTRACT
The research was carried out at the Bujoru Viticulture and Wine Growing Research and
Development Stage between 2015-2016. The paper presents the results of the research on vine
cultivation in ecological system. In the technology of vine cultivation, phytosanitary protection is
an important technological link for obtaining superior quantitative and qualitative productions.
The researches focused on the analysis of the disturbing factors in the ecological cultivation
system of the vine and the experimentation of how to minimize the anthropogenic intervention in
the ecological viticultural ecosystem. Pathogens were monitored by field observations and
notations to determine the frequency (F), intensity (I) and attack rate (G.A%). The recorded data
were correlated with the climatic conditions specific to the Dealul Bujorului vineyard.

INTRODUCTION
After entry of pests and diseases in the Americas growing countries in Europe,
growers, both practitioners and researchers alike have sought ways and means to
prevent and fight as effectively. Achieve high yields and good quality viticulture
involves the correct and optimal timing of all technological links, where it occupies an
important place against pathogens and pests.
The emergence and evolution of the main pathogens and pests of wine grapes
in the vineyard area Dealu Bujorului is influenced by direct and indirect effects of
technological and ecological factors specific area, affecting the quality and quantity of
grape production (Mirică I. et at, 1976; Tabaranu G. et al, 2005, 2007,2016;
Teodorescu Georgeta et at, 2003).

MATERIAL AND METHODS

The researches were conducted at the Research and Development Station for
Viticulture and Winemaking Bujoru in a vineyard cultivated with Merlot and Feteasca
regala aged 37 years.
Location experiment was conducted on land with a slope of 3-5% chernozem
soil type with a humus content between 1.14 to 1.86% in the A horizon and a weak
alkaline reaction (pH 7,44 to 8,30 ) and sandy loam; exhibition eastern land, about 140-
170 m altitude, orientation north-south rows, planting distance of 2,1 m x 1,2 m, density
provided 3968 vines / hectare Berlandieri x Riparia rootstock selection Telecky
Openheim SO4-4.
The organization experience of comprised the following varieties:

295
  Merlot - ecological technology ;
 Feteasca regala - ecological technology;
 Merlot - conventional technology;
 Feteasca regala - conventional technology
For the determine the gravity of the attack produced by the main vine diseases
of (manna, powdery mildew, gray rot) have been commented on the intensity (I) the
frequency (F) and degree of attack (AD/GA%) of their leaves and grapevine.

RESULTS AND DISCUSSIONS

Research has focused on technology experience of growing vines in the
ecological system, analyze quality grape production obtained and tested for efficacy
using various techniques and products to control pathogens and pests in organic
vineyards. To characterize the specific vineyard microclimate conditions Dealu
Bujorului were used weather data recorded at the meteorological station at Research
Station and Development for Vineyard and Vinification Bujoru.
From the analysis of the main factors climate during the growing season
(dezmugurit - defoliation) compared to multi-media mode (thermal, fluid, air humidity,
insolation), show that the condition of their vegetative development, production, its
quality and aggression of pathogens. Average temperatures in most months of the
vegetation period were close to normal, with the exception of April when average
temperatures were recorded higher than normal values (Table 1).

Table 1
Thermal regime during April – September 2016

The monthy Amount degrees of temperature

average
Global Active Effective
Months, temperature
year the
IV-IX the IV-IX the IV-IX the IV-IX
norm
2016 normal 2016 normal 2016 normal 2016
al
IV. 2016 11.7 13.0 349.5 389.8 278.8 338.3 78.06 98.3
V. 2016 18.1 15.3 560.4 473.4 555,9 473,4 257,5 16,4
VI. 2016 21,8 21,3 654,1 639,3 654,1 639,3 353,9 339,3
VII. 2016 24,1 22,9 744,2 709,2 744,2 709,2 435,0 399,2
VIII. 2016 23,1 22,3 698,6 690,5 716,7 690,5 406,6 380,5
IX. 2016 17,5 17,6 525,6 528,3 523,7 519,1 226,2 229,1

Highest temperature was recorded in August and 35.8°C in air. It is also worth
noting that in these months, there was a large number of days with maximum
temperatures above 30°C (75 days) as follows: 14 days in June, 24 days July 23 days
in August and 14 days in September. September was warmer than normal, the
average temperature was 17.6°C to 17.5°C annual average. The amounts degrees of
global temperature (global Σ°C), active (active Σ°C) and output (Σ useful °C) during the
growing season were much lower than the multi-values (Table 1). Thus, the entire
vegetation period of the overall heat balance was 3430.5°C to 3532.3°C multi-value,
the active heat balance was 3369.8°C to 3473.4°C, and useful thermal balance, was of
1462.8°C to 1757.26°C. Rainfall recorded during the growing season have been very
unevenly distributed, so were months when there have been very small amounts, well
below the normal value, such as July and August (Table 2) and Monday the s-
recorded greater amounts than normal: April, May (63.4 mm to 48.0 mm) and

296
June. The amount of rainfall during the growing season (April-September) was 319.0
mm against the normal 256.0 mm SCDVV Bujoru (Table 2).
Table 2
Weather data from the period april - september 2016

The number
Precipitations Hygroscopicity Heatstroke
of days with
Months, (mm) (%) (hours)
rain
year
the IV-IX the IV-IX the IV-IX
>10
normal 2016 normal 2016 normal 2016
IV. 2016 36.5 66.2 3 67.3 68.4 162.4 181.1
V. 2016 48.0 63.4 3 64.3 70.2 232.7 226.6
VI. 2016 72.1 74.2 2 64.6 70.0 236.0 238.1
VII. 2016 50.5 12.2 1 62.4 57.7 266.2 333.0
VIII. 2016 48.6 38.0 1 63.0 58.9 246.0 274.6
IX. 2016 40.3 65.0 2 69.3 63.1 172.4 196.1

Under these conditions and relative air humidity values were much higher than
the annual average 68.4% in April, 70.2% in May and 70.0% in June (Table
2). Heatstroke measured by the number of hours of sunshine was higher than normal
in all months during the growing season, with a total of 1449.5 hours of 1315.7 hours
SCDVV Bujoru normal value. Regarding risk factors during the growing season, we
can say that there was a total of 75 days with maximum temperatures above 30°C, the
combined amount of low rainfall in July and August led installing phenomenon drought
soil and air.Achieving high yields and good quality in viticulture, requires proper and
timely implementation of all links technology, in which an important place occupied by
combating pathogens and pests. In developing regimen for green variants took into
account the sensitivity of the two varieties studied, the economic damage threshold,
and restrictions imposed by legislation (Table 3).
In the climatic conditions of 2016 have been alerted and made seven
treatments to combat mildew of the vine, 6 treatments to combat and vine downy
mildew traps 3-6 / synthetic sex pheromone generation type for the control of grape
moth Atrabot (Figure 1).

Figure 1. Traps with pheromones type Atrabot

The vines manna (Plasmopara viticola - Berk. Et Curt.): favorable weather

conditions (precipitation frequency, high air humidity, etc.) have the appearance and
development of the pathogen to the compaction of the bloom clusters.In this case,
fungicides used were protected vines, limiting the degree of attack (GA%).Thus, Merlot
and Fetească regală - the degree of attack organic solution was slightly higher
compared to conventional alternatives, both on the leaves and grapes (Table 5).

297
 Table 3
Phytosanitary treatments applied in ecological technology - 2016

Date of Phenological Pathogens and Plant protection product Dose

treatment stage pests controlled use Kg., lt./ha
Sulfomat 80 PU
Oidium 4,00
19 april Shoots 2-6 cm Traps with pheromones
Eudemis G a I-a 3 buc.
type Atrabot
Manna Champ 77 WG 3,00
11 may Shoots 30-50 cm
Oidium Sulfomat 93 P 10,00
Manna Champ 77 WG 3,00
31 may Before blooming
Oidium Sulfomat 93 P 10,00
Bouille bordelaise
Manna 5,00
Immediately after Sulfomat 93 P
17 june Oidium 12,00
flowering Traps with pheromones
Eudemis G a-II-a 6,0 buc.
type Atrabot
Manna Bouille bordelaise 5,00
Growing shoots,
27 june
grains as peas
Oidium Sulfomat 93 P 12,00
Manna Bouille bordelaise 5,00
10 july Berries growing
Oidium Sulfomat 93 P 15,00
Manna Bouille bordelaise
5,00
Compaction of Oidium Sulfomat 93 P
25 july 15,00
grape bunches Eudemis G a-III-a Traps with pheromones
6,0 buc.
type Atrabot

Table 4
Phytosanitary treatments applied in conventional technology - 2016

Pathogens
Date of Phenological Plant protection Dose
and pests
treatment stage product use Kg., lt./ha
controlled
4,00
19 april Oidium Sulfomat 80 PU
Shoots 2-6 cm
Manna Dithane M 45 2,00
11 may Shoots 30-50 cm
Oidium Sulfomat 93 P 10,00
Manna Mikal Flash 3,00
31 may Before blooming
Oidium Sulfomat 93 P 10,00
Immediately after Manna Verita 2,50
17 june
flowering Oidium Bumper 250 EC 0,20
Manna Cupertine Super 4,00
Growing shoots,
27 june Oidium Falcon 460 EC 0,30
grains as peas
Sulfomat 93 P 10,00
Manna Champ 77WG 3,00
10 july Berries growing
Oidium Sulfomat 93 P 15,00
Compaction of grape Manna Bouille bordelaise 5,00
25 july
bunches Oidium Sulfomat 93 P 15,00

Grapevine powdery mildew (Uncinula necator - Schw. Burr.): powdery

mildew of vines degree of attack (GA%) amounts similar in all experimental versions
embodiment except for the green - royal Fetească which records a 13.97% AD% on
grapes (Table 5).
Gray rot of grapes (Botrytis cinerea - Pers.): Due to adverse climatic
conditions in 2016 (precipitations low from summer period, low atmospheric humidity)
symptoms pathogen attacks not been signaled in experimental groups.
298
 Table 5
Evolution of the main diseases in the year 2016

The degree of attack (%)

Varieties, technology Manna Oidium Gray rot
leaves grapes leaves grapes grapes
Merlot - ecological technology
3.17 16.64 0.46 3.04 0.0
Feteasca regala - ecological
technology 2.08 4.26 4.01 13.97 0.0
Merlot - conventional
technology 0.33 3.57 0.46 5.67 0.0
Feteasca regala - conventional
technology 0.94 0.94 3.87 3.30 0.59

Table 6
Evolution of grape moth (Lobesia botrana) in the year 2016

The degree of attack (%)

Varieties, technology The generation The generation The generation
a I-a a-II-a a-III-a
Merlot - ecological 0.33 0.0 3.66
technology
Feteasca regala - 0.33 4.00 7.00
ecological technology
Merlot - conventional 2.33 4.00 8.00
technology
Feteasca regala - 0.66 5.00 11.66
conventional technology

Grape moth (Lobesia botrana-Den et Schiff): in the period analyzed the

behavior and potential vine pest attack was highly influenced by climatic conditions
(temperature, humidity etc.).Temperature has a major influence on the development of
grape moth, such as: high temperature > 34-36°C are lethal for the eggs and adults,
eggs and larvae resistant to temperatures between 0°C and 10°C and to 23°C
crisalidele. Following the observations made on the varieties in the experimental lots,
the grape moth recorded a lower degree of attack (G.A%) in all three generations
compared to conventional technology (Table 6).

CONCLUSIONS
 Analysis of the climatic elements of 2016 highlights the fact that in the period
from May to June were favorable conditions for the development of developing
and hands, and during July-August for mildew.
 Phytosanitary treatments applied to ecological technology to combat
pathogens and pests vines in specific climatic conditions of 2016, highlights
that plant protection products applied at the recommended dose and the
optimal time protects the vines.

ACKNOWLEDGMENT
This work was conducted under the project: ADER 3.3.8 .: Adaptation
technologies of cultivation of the vine in the ecological system, maximizing resource
biotic and abiotic ecosystem wine, in order to preserve biodiversity.

299
 REFERENCES
Mirică I. şi Mirică Afrodita,1976 - Combaterea bolilor şi dăunătorilor la viţa de vie,
Editura Ceres, p 25 - 55.
Teodorescu Georgeta, Roman T. şi Mirică Sumedrea Mihaela, 2003 - Entomologie
Horticolă, Editura Ceres,Bucureşti p 330 - 343.
Ulea E., 2003 - Fitopatologie, Ed."Ion Ionescu de la Brad", Iaşi, p 205 - 229.
Simion Cristina, 2003 - Tehnologie ecologică de producere a strugurilor pentru
obţinerea vinurilor roşii. Ed. Pax Aura Mundi, Galaţi, 27-30.
Tabaranu G., Simion Cristina, 2005 - Întrebări şi răspunsuri privind recunoaşterea
şi combaterea dăunătorilor precum şi a celor mai frecvente boli în plantaţiile viticole din
S-E Moldovei, Editura Pax Aura Mundi, Galaţi.
Tabaranu G, Enache viorica, 2016 - Researches on of grape moth the evolution
(Lobesia botrana - Den. et Schiff.) of Dealul Bujorului the vineyard, in the context of
current climate change. Lucrări ştiinţifice, sr. Horticultură, vol. 59 (1/2) 2016
*** Ministerul Agriculturii şi Industriei Alimentare, 1980 - Metodici de prognoză şi
avertizare a tratamentelor împotriva bolilor şi dăunătorilor plantelor de cultură ,
Bucureşti.

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ANNALS OF T HE  Horticu lture
 Food produce process ing
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techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

RESEARCH ON OBTAINING FRESH CHEESE FROM GOAT'S MILK WITH

TARRAGON

Tiţa Mihaela-Adriana1*, Popovici Cristina2

1*
”Lucian Blaga” University of Sibiu, Faculty of Agricultural Sciences, Food Industry and Environmental
Protection
2
Technical University of Moldova, Faculty of Food Technology, Chisinau, Republic of Moldova
*Correspondence author. E-mail: [email protected]

Key words: cheese, tarragon, innovative, bioactive components

ABSTRACT
This study aims at developing an innovative dairy product – cheeses, which besides the
benefit of the existing native ingredients in the raw material – goat’s milk, it is enriched with
natural products of vegetable origin with valuable bioactive principles. The natural product of
vegetable origin - tarragon, comes from the local flora, being rich in volatile oils, aromatic
compounds, antioxidants, minerals, vitamins. Have been realized three samples of cheese that
contain different quantities of tarragon and have been analyzed over a period of 6 days from a
sensory and physicochemical point of view.

INTRODUCTION
In recent years, many cheeses (C.Banu şi Vizireanu,1998) have responded to
consumer demands of various varieties by adding flavor ingredients, herbal
supplements such as parsley or chives, garlic, pepper or cumin, black pepper.
Aromatic plants are a source of biologically active compounds which, introduced into
food, can give them primarily the flavor of the volatile oils they contain, secondly they
can improve their color through anthocyanins and flavonoids, and last but not least
increases the antioxidant capacity of foods through their own vitamins, polyphenols or
flavonoids (Acharya Sn, Thomas Je, Basu Sk. ,2007). A strong aromatic plant that
draws its origins from Mongolia and Siberia, the tarragon (Arthemisia Dracunculus)
was introduced in Europe in the 16th century. In the chemical composition of tarragon
are volatile oils, aromatic compounds, antioxidants, minerals, vitamins, tannins, bitter
substances, iodine, mucilages, substances giving the plant carminative, diuretic,
expectorant, calming, digestive, aperitive, antiparasitic, antispasmodic, antiviral,
decongestant, antiinfectious, disinfectant, anti-inflammatory and flavoring proprieties.
The tarragon is a plant rich in vitamins, such as vitamin C, vitamin A, B complex
vitamins, folic acid, pyridoxine, niacin, riboflavin, etc., which function as antioxidants as
well as co-factors of metabolism. The tarragon is especially an excellent source of
minerals, such as calcium, manganese, iron, magnesium, copper, potassium and zinc.
The tarragon is widely used in cooking and flavor, but little of us know that it is also
effective in medicine. It is a key plant in French cuisine (it is an essential ingredient in
Bearnaise sauce) and works very well with egg, cheese. In our country, it is known as
essential spice in the Ardeal cuisine and is usually used in dry form, although the taste
of fresh tarragon is much more interesting and more armatically than dry.
301
 MATERIAL AND METHODS
In this research we used goat's milk from a private farm, which has a
corresponding microbiological quality, being processed into fresh cheese according to
a traditional recipe. After the squeeze process, we made three samples of fresh
cheese (Costin Gh. M.,2003) with a different addition of tarragon, which was previously
washed, finely chopped and mixed with a little salt. The samples contain 200 grams of
cheese with tarragon and was noted like this: PB1 - containing an addition of 10 grams
of tarragon, PB2 - 15 grams of tarragon, PB3 - 20 grams of tarragon. These samples
were analyzed over a period of six days in sensory and physicochemical terms (Tita M,
2002). Sensory analysis - using the method of comparison with unit scoring scales,
which consists of assessing each sensory characteristic and obtaining a medium score
given by 3 tasters. The score for assessing the sensory characteristics was as follows:
5 points - very good; 4 points - good; 3 points - satisfactory; 2 points - unsatisfactory; 1
point - inappropriate; 0 points - altered; Determination of dry substance - Moisture
analyzer AND ML-50 is based on the principle of thermogravimetric analysis, drying of
the samples using the halogen lamp and obtaining the moisture content in %;
Determination of lactic acid - Semiautomatic Clinical Biochemistry Analyzer State Fax
1904, expressed in grams/100 grams product; Determination of water activity using the
"Novasina" apparatus.

RESULTS AND DISCUSSIONS

a. Sensory analysis: The tasters analyzed the sensory characteristics of
cheese with tarragon for a period of 14 days, by comparing with scoring ranges from 0
to 5 points, established in sensory analysis standards. The scores and also the ratings
given by the tasters for this type of cheese are shown in table no.1.
Table 1
Sensory characteristics of cheese with tarragon
Samples Day 1 Day 3 Day 6
analyzed Scores Qualifying Scores Qualifying Scores Qualifying
Sample 1 4 B 3 S 2 NS
Sample 2 5 FB 4 B 4 B
Sample 3 3 S 2 NS 2 NS
Note: FB – very good; B – good; S – satisfactorily; NS – unsatisfactory; N – improperly;
A – adulterated

From the results obtained, we can see that the PB2 sample obtained the best
scores and qualifiers, which denotes the content of tarragon in the cheese is optimal,
compared to the other samples.
b. Determination of dry substance at thermobalance: The results obtained in
determining the dry substance are shown in figure no.1.
From the results presented, it is observed that the three samples subjected to
the analysis show slight increases of the dry substance over the analyzed period, and
the different content of the dry substance of the three samples is given by the amount
of tarragon that differs.

302
 Figure no.1. Evolution of dry matter

c. Determination of lactic acid: The results obtained in determination of lactic

acid are shown in figure no. 2

Figure no. 2. Evolution of lactic acid

It is noted that the lactic acid content increases as a result of lactic acid activity
on lactose throughout the analyzed period. The lactic acid content is also influenced by
the amount of tarragon, so sample PB1 has the highest amount of lactic acid and PB3
has a lower amount of lactic acid. For this reason, the shelf life of PB3 is higher than
that of PB2.
d. Determination of water activity: The result obtained in determination of water
activity are shown in figure no. 3
Water activity is the result of processes that take place during the analyzed
period and is seen to increase due to the fact that a balance is achieved between the
components of the products, and from the data presented, it is observed that the water
activity does not show any significant differences during the analyzed period, which
represents a safety of tarragon cheeses samples from a microbiological point of view.

303
 Figure no. 3. Evolution of water activity

CONCLUSIONS
Following the analysis of the three samples of fresh tarragon cheese during the
analyzed period, the following conclusions can be drawn:
 PB2 on sensory analysis has received the highest scores and ratings, which
means it has the optimal content of tarragon;
 The content of dry substance and lactic acid is influenced by the amount of
tarragon added;
 The activity of water does not show significant differences during the analyzed
period, which is a safety of tarragon cheeses samples from a microbiological
point of view.

REFERENCES
Acharya Sn, Thomas Je, Basu Sk. ,2007, Methods for the improvement of plant
medicinal properties with particular reference to fenugreek (Trigonella foenumgraecum
L.). In: Acharya SN, Thomas JE, Eds. Advances in medicinal plant research. India:
Research Signpost Chapter, pag. 491-512;
C. Banu şi Camelia Vizireanu, 1998,Procesarea industrială a laptelui – Editura
Tehnică,Bucureşti;
Costin Gh. M.,2003, Ştiinţa şi ingineria fabricării brânzeturilor, Editura Academica,
Galaţi,;
Tiţa Mihaela – 2002, „Manual de analiză şi controlul calităţii în industria laptelui”,
Editura Universităţii „Lucian Blaga” din Sibiu.

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ANNALS OF T HE  Horticu lture
 Food produce process ing
UNIVERSITY OF CRAIO VA
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

THERMAL ANALYSIS OF CURD OBTAINED BY DIFFERENT METHODS

1* 2
Tiţa Mihaela-Adriana , Popovici Cristina
1*
”Lucian Blaga” University of Sibiu, Faculty of Agricultural Sciences, Food Industry and Environmental
Protection
2
Technical University of Moldova, Faculty of Food Technology, Chisinau, Republic of Moldova
*Correspondence author. E-mail: [email protected]

Key words: coagulation process, quality, maturation, differential scanning calorymetry

ABSTRACT
To obtain high-quality cheese is dependent of the coagulation mode, which can be
about acids or enzymes. A coagulation process that respects the technological parameters
afford the obtaining of a good quality curd. In our study we used goat milk which was coagulated
with enzymes using the rennet and the coagulated with acid using citric acid, acetic acid. The
resulting curd was analyzed over a period of 14 days using differential scanning calorimetry
(DSC). Following the analysis performed by differential scanning calorimetry were determined
melting temperatures of the three types of curd, taking into account the time in which melting
occurs. The results obtained showed that the highest temperatures melting occurred at the curd
obtained by adding rennet, which has a specific consistency and allows to obtain a high quality
cheese, both in terms of sensory and physico-chemical.

INTRODUCTION
Improved coagulation properties of milk from individual goats (C. Banu, et.al.,
2007) were shown to be associated with a higher content of total and colloidal calcium
to some extent, low casein micelles. Cold storage has been shown to induce RCT
longer for both types of milk after 24 hours, where the gel strength was reduced
primarily in good coagulation of milk samples during storage. Milk, cold storage for up
to 14 hours did not affect the rheological, although RCT tended to decrease after
pasteurization HTST. Further studies are needed to understand how milk processing
steps affecting the distribution of casein and colloidalmineral phases between
individual serum and milk coagulation with different properties. In addition,
pasteurization HTST currently applied has no significant effect on coagulation
properties, even if RCT was reduced by about 7 min. This is in contrast to earlier
studies, where pasteurization of the cheese making properties improved by partially
reverse micelle dissociation of Ca, P inorganic casein occur during cold storage (Ali
AE, et.al.1980). Differences in developing coagulation properties could be attributed to
changes affecting the second stage coagulation. In this way, more investigation is
needed to characterize the relationship between the second stage of coagulation
induced chymosin and casein micelles changes occurring during storage in cold milk
coagulation distinct individual skills. Technological intervention milk through the
production line cheese (ex: cold storage, standardization fat, pasteurising, CaCl 2
addition and acidification) can have a significant impact on the physico-chemical in milk

305
and therefore on its properties and the rheological characteristics of the cheese.(De la
Fuente M,1988).
It is known that the pH affects the rate of renneting, curd firmness and
syneresis and the recovery of fat, the retention of calcium and, therefore, final cheese
texture.(Watkinson P,et.al.,2001). However, the stored cold pasteurization of milk is
known to recover its coagulation properties in a certain extent by reversing the above-
mentioned changes in the structure of the casein micelle. (Frederiksen PD,
et.al.,2011). Chymosin induced coagulation is a two-step process. The first step is κ-
CN chymosin cleavage in para-κ-CN and casein macropeptides (CMP). To about 85%
of ĸ-CN it was cleaved to promote the formation of aggregates. An increase in viscosity
is observed followed by branching of the aggregates, as characterized the second
stage. (Dalgleish DGD, Corredig M, 2012). It was questioned whether molecules of κ-
CN milk non coagulants are actually split in the same way as the κ-CN good milk
coagulant chymosin, but in a previous study, it was observed that the first phase held
at comparable as far as good and non coagulating. (Frederiksen PD, et.al.,2011).

MATERIAL AND METHODS

The milk used to obtain different types of clot is the goat originated from a farm
in the county of Sibiu. Milk has undergone technological operations reception,
pasteurization and cooling (.
Differential scanning calorimetry (DSC)
Thermal analysis (TA) is a group of techniques in which changes of physical or
chemical properties of the sample are monitored against time or temperature, while the
temperature of the sample is programmed. The temperature program may involve
heating or cooling at a fixed rate, holding the temperature constant (isothermal), or any
sequence of these. Determines: melting – crystallization; polymorphism/phase
diagram; glass transition; degree of crystallinity of semi-crystalline material; degree of
curing/cross linking; specific heat capacity (J/gK;) decomposition onset; reaction
kinetics, enthalpy; oxygen resistance (oxidative stability, OIT); purity determination.
The characteristics thermal analysis: samples can be analyzed in a wide range of
temperatures, using various temperature programs, dynamic and isothermal
conditions; samples can be in any physical form (solid, liquid, gel) in different forms
(powder, films, granules, fibers, fabrics, plate, etc.); it is necessary a small amount of
sample (0.1 mg-10 mg); the time required for analysis varies from a few minutes to
several hours; atmosphere around the sample can be defined (air/inert gas);
measurement techniques and sample handling is not difficult; there are a variety of
tools AT commercially available, and the price is moderate.
Differential scanning calorimetry (DSC) (G. Bruylants, et.al, 2005) is a
technique of recording energy required to maintain a zero temperature difference
between the sample cup and the cup is the reference, both of which are
simultaneously heated or cooled at a controlled rate. If the sample undergoes a
transition, the endo- or exothermic process it or reference cup will receive an
equivalent energy so as to maintain the temperature equal to two cups (reference
sample respectively).

RESULTS AND DISCUSSIONS

Slow heating rate leads to a balance between the reactions taking place. At
first the temperature is high, the sample undergoes a glass transition (glass), then at a
certain temperature and produce heat crystallized sample is an exothermic process
taking place, so at the lowest recorded temperature crystallization. Heating the
samples in turn heats the crystalline areas is a process of melting, i.e. an endothermic
306
process, so that the highest peak indicates the melting temperature of the sample. For
each sample analyzed at T0, T7 and T14 the two thermogram were obtained in the first
are the melting temperature of the test sample, and in the second time at which the
melting of the sample. The results obtained are presented in the following figures:

Fig.1. Analysis curds obtained by adding rennet.Melting temperature

Fig.2. Analysis curds obtained by adding rennet. Melting time.

The melting temperature for the sample T0 is 110°C to 8 minutes at 100°C T 7
is 7 minutes at 105°C T14 is 7.5 minutes. Thus the highest melting temperature was
recorded at T0, and the lowest at T7.

307
 Fig.3. Analysis curds obtained by adding acetic acid. Melting temperature.

Fig.4. Analysis curds obtained by adding acetic acid. Melting time

The melting temperature for the sample T0 is 97°C 6.8 minutes at 105°C T7 is
7.5 minutes at 102°C T14 is 7.3 minutes. Thus the highest melting temperature was
recorded at T7, and the lowest at T0.

308
 Fig.5. Analysis curds obtained by adding citric acid. Melting temperature.

Fig.6. Analysis curds obtained by adding citric acid. Melting time.

The melting temperature for the sample T0 is 103°C, 7.3 minutes at 96°C T7 is
6.6 minutes at 100°C T14 is 7.0 minutes. Thus the highest melting temperature was
recorded at T0, and the lowest at T7.

309
 CONCLUSIONS
This study showed that the analyzes performed were determined by DSC
melting temperatures of the four types of curd, as was revealed that the highest
melting temperatures were recorded at curd obtained by adding rennet, which has a
specific consistency and allows to obtain a high quality cheese, both in terms of
sensory and physico-chemical.

REFERENCES
Ali AE, Andrews AT, Cheeseman GC,1980, Influence of storage of milk on
casein distribution between the micellar and soluble phases and its relation ship to
cheese-making parameters, Journal of Dairy Research, 47, p.371-382;
De la Fuente M,1988, Changes in the mineral balance of milk submitted to
technological treatments, Trends in Food Science and Technology 9, pp.281–288;
Watkinson P, Coker C, Crawford R, Dodds C, Johnston K, McKenna A, White
N,2001, Effect of cheese pH and ripening time on model cheese textural properties
and proteolysis., International Dairy Journal,11, pp.455–464;
Dalgleish DGD, Corredig M, 2012,The structure of the casein micelle of milk and
its changes during processing, Annual Review of Food Science and Technology, 3,
pp.449–467;
Frederiksen PD, Hammershøj M, Bakman M, Andersen PN, Andersen JB, Qvist
KB, Larsen LB, 2011,Variations in coagulation properties of cheese milk from three
Danish dairy breeds as determined by a new free oscillation rheometry-based
method, Dairy Science and Technology, 91, pp.309–321;
C. Banu, Gh, Păsat, S. S: Dorin, Aura Darabă,2007, Valorificarea laptelui de
capră. Ghid pentru fermieri, Editura Agir, p.68, 71,138-143;
G. Bruylants, J.Wouters and C.Michaux, 2005 ,Differential Scanning Calorimetry
in Life Science: Thermodynamics, Stability, Molecular Recognition and Application in
Drug Design, Current Medicinal Chemistry, 12, pp.2011-2020.

310
 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
 Food produce process ing
UNIVERSITY OF CRAIO VA
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

USING THE BIOACTIVE POTENTIAL OF PLANTS BY GETTING TONIC

DRINKS

Tița Ovidiu1*
1
“Lucian Blaga” University of Sibiu, Victoriei Boulevard, no. 10, Sibiu, Romania
*
Corresponding author, e-mail: [email protected]

Key words: potentially bioactive, tonifying, plants, extract, wine

ABSTRACT
The paper presents important issues regarding the possibility of using plants with
bioactive potential recognized in the production of tonic drinks with a beneficial role in human
health. Bioactive plants are currently used both for medical purposes and for the production of
alcoholic beverages, extracts or tinctures. We can use plants that grow very well in our country
and are the subject of important crops for export. Diversifying their use can have beneficial
effects. The use of these in everyday life in various forms ensures the natural resistance of the
organism to some contamination factors such as bacteria or fungi.

INTRODUCTION
The beneficial effects of plant extracts in healing or ameliorating diseases have
long been used in folk medicine (Tabata et al., 1988). The properties of these plants
are based on their rich composition of bioactive compounds such as essential oils,
tannins, flavones or polyphenols. Terpene, monoterpene, diterpene, sesquiterpene,
these have antibacterial, antiseptic, antiinflammatory or antiviral properties (Wallace,
2004). Antimicrobial activity is due to compounds such as carvacol, camphor or thymol,
the activity of which is beneficial to the human body, having uses in the pharmaceutical
or food industry (Bhaskara et al., 1998; Ali-Shtayeh et al. 2008; Hadizadeh et al. 2009).
Plant aromas lead to an olfactory perception, based on higher alcohols, terpenic
compounds, bitter substances, aldehydes and ketones, esters, and quantitative and
qualitative quantification is achieved by modern methods such as gas chromatography
(Târdea, 2010). Polyphenols are natural compounds found in herbal foods, such as
fruits, vegetables, whole grains, tea, wine, cocoa, etc. (Pandey & Rizvi, 2009).

MATERIAL AND METHODS

The plants with potentially bioactive potential used were: aphid (leaves and
fruits) (Vaccinium myrtillus L.), rosehip (Rosa canina L.), cranberry (Vaccinium vitis-
idaea L.), blackberries (Rubus fruticosus L. ), wormwood (Artemisia absinthium L.),
zmeur (leaves and fruits) (Rubus idaeus L.) which have been dried and milled. The
obtained powder was homogenized in 50% alcoholic solution in order to extract the
bioactive principles in a ratio of 1:10 for 24 hours. The samples were centrifuged and
the supernatant was used in the assays. Evaluation of aromatic compounds in plants
was performed using the GC / FID system (gas chromatograph coupled with a flame
ionization detector. Maceration of plants to obtain plant maceration was performed at
311
40-450 ° C with a 45-50% vol. Hydroalcoholic solution for 18 hours. The plant / solvent
ratio was selected to 1/10.
The ratio of the weight of the 6 plants used to obtain the maceration is
confidential. Specialty studies on red wines show that their composition and properties
can be a basis for the production of tonic beverages. Wines of the Fetească Neagră
variety are indigenous wines with physicochemical properties and special sensors, well
appreciated on the domestic and foreign market, and due to these attributes it is also
suitable for use in various recipe for toning drinks. A wine of this variety from the 2015
harvest year was used in the Dealul Mare Vineyard with the following characteristics:
reducing sugar 3.7 g / L, pH 3.8, total acidity of 4.1 g / L tartaric acid and alcoholic
strength of 12.5% v / v.

RESULTS AND DISCUSSIONS

In the realization of the herbal tonic drink, the specific stages of raw materials
analysis, wine, sugar, refined food alcohols, aromatic plants, conditioning of the used
raw material, preparation of sugar syrup and aromatic herbs macerate were covered.
We then proceeded to the actual realization of the tonifying drink that remained in the
twinning of the components for 30 days of 15-180C before carrying out physico-
chemical and sensory analyzes.
They have been identified as particularly valuable wine compounds: 1431.257
mg / L of total polyphenols, of which 488.137 mg / l tannin, 369.027 mg / L
anthocyanins and 101 271 mg / L of phenolic acids.
A series of phenolic compounds were also identified by HPLC methods:
quercetin 0.5215 mg / 100 mL vin, gallic acid 8.8147 mg / 100 mL vin, acaric syrup with
1.9957 mg / 100 mL vin, caffeic acid with 4.17975 mg / 100 mL of wine. Resveratrol
was identified in the amount of 0.9173 mg / 100 mL of wine.

Figure 1. Sensory analysis for Black Fetească wine

312
 Figure 2. Sensory Analysis 1 for Tonic drink

Figure 3. Sensory Analysis 2 for Tonic drink

The not very high acidity of the wine, its aroma and its softness cover the
astringent of the plant extract used to make the toning drink.
As it results from the sensory analysis of the Black Feteasca wine, it is
distinguished by its typical character, it has a pronounced floral character, the mineral
character being more discreet. It has an excellent taste balance and a very good
persistence of taste. It has a pleasant astringency rated with a score that can print
harmony and ensures a pleasant tonic drink.
A first sensory analysis of the tonic drink obtained is presented as a product
with a strong taste of wormwood, but also of dried flowers and fruits, keeping very well
the basic material from which the product originates. It has a complex, pleasant,
313
harmonious taste. The parameters analyzed for the second sensory analysis attest to
the achievement of a harmonious, good-tasting product, a persistent pleasant,
refreshing taste without a astringency that would lower its value.

CONCLUSIONS
The tonic beverages have a small caloric intake, they help digestion, contain
plant compounds with high antioxidant potential and are recommended for various
diseases or as digestive and toning drinks. Polyphenols from these herbal drinks
based on red vines have a pronounced antiviral, antibacterial and protective role of the
body against atherosclerosis, facilitating biochemical reactions that help protect the
cardiovascular system. In addition to the antioxidant effect, it promotes vitamin C
action, helps reduce cholesterol, and resveratrol has the role of stopping the spread of
cancer cells. The content of polyphenols, tannins, flavones, vitamins, minerals or
phenolic acids give rise to specific flavors, which ultimately contribute to achieving the
particularly pleasant sensory aspect of the toning product obtained.

REFERENCES
Ali-Shtayeh M.S., Jamous R.M, Al-Shafie J.H., Elgharabah W.A., 2008. Traditional
knowledge of wild edible plants used in Palestine(Northern West Bank): A comparative
study, J. Ethnobio. Ethnomed., doi: 10.1186/1746-4269-4-13.
Bhaskara M.V., Angers P., Gosseline A., Arul J., 1998. Characterization and use
of essential oil from Thymus vulgaris against Botrytis cinerea and Rhizopus stolonnifer
in strawberry fruits,Phytochem., 47, 1515- 1520.
Hadizadeh I., Peivastegan B., Hamzehzarghani H., 2009. Antifungal activity of
essential oils from some medicinal plants of Iran against Alternaria alternate, Am. J.
Appl. sci., 6, 857-861.
Pandey K.B., Rizvi S.I., 2009. Plant polyphenols as dietary antioxidants in human
health and disease. Oxid Med Cell Longev., 2:270–278.
Tabata M., Honda G., Sezik E., 1988. A report on traditional medicine and
medicinal plants in Turkey. Faculty of Pharmaceutical sciences, KyotoUniversity.
Țârdea C., Sârbu Gh., Țârdea A., 2010. Tratat de vinificație, ed. Ion Ionescu de la
Brad, Iași, pp 725-734
Wallace R.J., 2004. Antimicrobial properties of plant secondary metabolites, Proc.
Nutr. Soc., 63, 621-629.

314
 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
 Food produce process ing
UNIVERSITY OF CRAIO VA
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

INFLUENCE OF FRUIT LOAD ON THE QUALITY OF GRAPE PRODUCTION

AND EVOLUTION OF PHENOLIC COMPOUNDS DURING THE
FERMENTATION MACERATION OF WINES IN DEALU MARE VINEYARD
1
Tudor Georgeta *, Pîrcălabu Liliana
1*
Research Institute for Viticulture and Enology Valea Calugareasca
* Correspondence author. E-mail: [email protected]

Key words: polyphenols, alcohol content, maceration-fermentation

ABSTRACT
The varieties taken into study were Feteasca neagra and Cabernet Sauvignon, part of
the basic assortment for the production of quality red wines, typicaly for Dealu Mare vineyard,
which are very valuable in view of the oenological aspect.
At harvest (19 September 2016) the grapes has sugar accumulations of 222 g/l
(Cabernet Sauvignon) and 226 g/l (Feteasca neagra). The total anthocyanin concentration was
1346 mg/l in case of Cabernet Sauvignon variety and 1267 mg/l at Feteasca neagra, values
closed to those of grapes rich in anthocyanins which have more than 1200 mg/l (Ribereau-
Gayon P., 1964, 1972). The extractibility of anthocyans (62% at Cabernet Sauvignon and 57% at
Feteasca neagra) and the maturity of the seeds (72% at Cabernet Sauvignon and 69% at
Feteasca neagra) were very good.

INTRODUCTION
The factors determining the alcohol content of wines are: varieties of vines; the
soil on which the vine is grown (calcareous soils generally giving the most alcohol-rich
wines); the climate of the vineyard and the way climatic factors evolve during the year;
technological conditions and especially fermentation of must (ŢARDEA, 2007; PATIC,
2006).
The evolution of phenolic compounds during the maturing of grapes for red
wines was determined and the amount of polyphenols in the skin of the berry grape is
10 to 100 times higher than in the must, increasing in the order of Feteasca neagra
and Cabernet Sauvignon, with large differences between varieties (KONTEK Adriana,
1975, 1983). The total amount of polyphenols in the skins is increasing after the grape
harvest, up to technological maturity and found in seeds, 150 times more than in the
must, and the flavonoly tannins are found in large quantities in the seeds (BOURZEIX
M., 1975, 1976).

MATERIAL AND METHODS

The research was carried out in two vineyards grown with Feteasca neagra
and Cabernet Sauvignon. The study was performed during 2016 harvest, with climatic
conditions favorable for the over-maturing of grapes. The cutting system practiced was
Guyot Multiple with 3 loadings: 20 eyes/vine (T1), 28 eyes/vine (Mt) and 36 eyes/vine
(T2), with the attribution of a cutting load of 8-10 eyes per vine and two eyes taps. nder
the conditions of microvinification, three experimental variants were made for each
315
selected vinifera variety, the differentiation being made according to the time of
separation of the marc, respectively 8 and 16 days.

RESULTS AND DISCUSSIONS

The experimental results obtained showed that under the ecoclimatic
conditions specific to 2016 year, characterized by a moderate heliothermic regime on
the background of rich water resources, the experimental technological variants had a
different impact on the quality of the grape production and wine. The lowest minimum
temperature was recorded in September (14.1°C) and the highest in July (18.3°C), the
lowest maximum temperature was in September (25.6°C) and the higher in July
(29.6°C), (Table 1).

Table 1
The thermal and pluviometric regime during June-September

Air temperature
Average Average Rainfalls Huglin
Luna Average
minimum minimum (mm) Index
temperature
temperature temperature
(oC)
(oC) (oC)
June 22.1 17.1 27.4 81.8 442.5
July 23.7 18.3 29.6 70.2 516.2
August 22.7 17.5 29.2 78.2 494.5
September 19.0 14.1 25.6 66.0 381.3

During the ripening and full maturation rainfall data indicate precipitation
amounts with a minimum of 66 mm in September and a maximum of 81.8 in June.
At harvest (19 September 2016) the grapes has sugar accumulations of 222 g/l
(Cabernet Sauvignon) and 226 g/l (Feteasca neagra). The total anthocyanin
concentration was 1346 mg/l in case of Cabernet Sauvignon variety and 1267 mg/l at
Feteasca neagra, values closed to those of grapes rich in anthocyanins which have
more than 1200 mg/l (Ribereau-Gayon P., 2012). The extractibility of anthocyans (62%
at Cabernet Sauvignon and 57% at Feteasca neagra) and the maturity of the seeds
(72% at Cabernet Sauvignon and 69% at Feteasca neagra) were very good.
The maceration-fermentation process began on 22 September 2016 and
ended on 7 October 2016. The dynamics of polyphenols extraction was continued until
the separation of the wine from marc. The amount of anthocyanins increased
progressively until day 7, and then decreased continuously until day 15, due to their
involvement in polyphenolic stabilization and absorption on the yeast.
The dynamics of anthocyanins during fermentation maceration is continued
until the time of separation of the marc (8th day of fermentation maceration),
respectively 436 mg/L in case of Feteasca neagra 20 eyes/vine and 317 mg/L to
Feteasca neagra 36 eyes/vine. The content polyphenols shows variations between
1809 mg/L (Feteasca neagra 20 eyes/vine) and 1605 mg/L (Feteasca neagra 36
eyes/vine), (Table 2).

316
 Table 2
The content of anthocyanins and polyphenols of the separated fractions at 8 and 16
days

a.Feteasca neagra
Anthocyanyns (mg/L) Total polyphenols (mg/L)
The maceration-
fermentation
36 eyes/ 28 eyes/ 20 eyes/ 36 eyes/ 28 eyes/ 20 eyes/
period
vines vines vines vines vines vines

8 days 317 375 436 1605 1655 1809

16 days 285 306 339 1587 1610 1649
b. Cabernet Sauvignon
Anthocyanyns (mg/L) Total polyphenols (mg/L)
The maceration-
fermentation
36 eyes/ 28 eyes/ 20 eyes/ 36 eyes/ 28 eyes/ 20 eyes/
period
vines vines vines vines vines vines

8 days 364 364 424 2081 2213 2567

16 days 343 350 404 2053 2334 2526

When separating the fractions to 16 days the content of anthocyanins is

continuously decreasing, respectively in the Feteasca neagra variety from 339 mg/L
(20 eyes/vine) to 285 mg/L 936 eyes/vine), and in the case of the Cabernet Sauvignon
variety from 404 mg/L (20 eyes/vine) to 343 mg/L (36 eyes/vine).
Extending the maceration-fermentation time from 8 to 16 days after the end of
alcoholic fermentation leads to important changes in the red wine structure: decreases
the total anthocyanin content and the color intensity of the wine; increases the content
in total polyphenols, modifying the indices characterizing the phenolic complex in wine,
namely:
The color intensity for Feteasca neagra variety (20 eyes/vine) from 8,350 (8
days) to 7,610 (16 days), 28 eyes/vine from 6,990 (8 days) to 6,370 (16 days), 36
eyes/vine from 6,970 (8 days) to 6,260 (16 days). The same variation is found for
Cabernet Sauvignon 20 eyes/vine from 11,350 (8 days) to 9,350 (16 days), 28
eyes/vine from 10,920 (8 days) to 8,850 (16 days) to 36 eyes/vine from 10,030 (8
days) to 8,740 (16 days).
The extension of the maceration-fermentation period induces an increase in
the total polyphenols content, as follows: in Feteasca neagra variety in case of 20
eyes/vine from 1585 (8 days) to 1783 (16 days) at 28 eyes/vine at 1580 (8 days) at
1612 (16 days) and at 36 eyes/vine from 1527 (8 days) to 1569 (16 days). And in the
Cabernet Sauvignon variety, increases in total polyphenols in 20 eyes/vine from 2441
(8 days) to 2500 (16 days), 28 eyes/vine from 2165 (8 days) to 2282 (16 days), and at
36 eyes from 1968 (8 days) to 2011 (16 days), (Table 3).
The sensory profile of the wine was obtained on the basis of a tasting score
using the red wine specific descriptors. The quality level of aromatic flavor components
was average for Feteasca neagra wine, harvest 2016, with the exception of the
caramel note that had 4 points for 20 eyes/vine -8 days. The amount of tannins was
average, perceived with a slight astringency in 20 eyes/vine and 36 eyes/vine (Figure 1
and Figure 2).

317
 Table 3
Physico-chemical characteristics of Feteasca neagra wines

Feteasca neagra
Variant/Parameters 28 eyes/vine 20 eyes/vine 36 eyes/vine
8 16 8 16 8 16
days days days days days days
Alcoholic concentration (% vol) 13.0 13.2 13.2 13.4 12.8 13.0
Sugars reducing (g/l) 4.85 4.96 4.03 4.07 5.20 5.30
Total acidity (g/l tartric acid) 5.79 5.45 5.49 5.27 6.09 6.17
Volatile acidity (g/l acetic acid) 0.63 0.67 0.62 0.64 0.65 0.67
Total extract (g/l) 23.7 24.2 22.7 22.9 24.8 28.7
Nereducator extract (g/l) 18.85 19.24 18.67 18.83 19.6 23.4
Color intensity (Ic) 6.990 6.370 8.350 7.610 6.970 6.260
Anthocyanins (520 nm) 347 298 425 334 309 278
Total polyphenols 1580 1612 1585 1783 1527 1569

Physico-chemical characteristics of Cabernet Sauvignon wines

Cabernet Sauvignon
Variant/Parameters 28 eyes/vine 20 eyes/vine 36 eyes/vine
8 16 8 16 8 16
days days days days days days
Alcoholic concentration (% vol) 12.5 12.8 12.7 12.8 12.0 12.4
Sugars reducing (g/l) 5.56 5.68 4.95 4.98 5.86 5.94
Total acidity (g/l tartric acid) 5.79 5.64 5.72 5.49 6.55 5.94
Volatile acidity (g/l acetic acid) 0.75 0.76 0.74 0.75 0.77 0.79
Total extract (g/l) 25.3 25.8 23.7 23.9 25.5 26.0
Nereducator extract (g/l) 19.74 20.12 18.75 18.92 19.64 20.06
Color intensity (Ic) 10.92 8.850 11.35 9.350 10.03 8.740
Anthocyanins (520 nm) 347 336 415 394 340 326
Total polyphenols 2165 2282 2441 2500 1968 2011

The Cabernet Sauvignon wine has a reddish color with violet shades and a
gusto-olfactory herbaceous character. The dominant feature is blackcurrant and bitter
chocolate (20 eyes/vine - 8 days) and the amount of tannin was maximum in 20
eyes/vine - 16 days.

318
 Aromatic
intensity
4
Quality tannins 3 Aromatic quality

2
1
Quantity tannins Dried prunes
0

Astringent Black currants

Caramel Rose

28 eyes/vine- 8 days 28 eyes/vine- 16 days

20 eyes/vine- 8 days 20 eyes/vine- 16 days
36 eyes/vine- 8 days 36 eyes/vine- 16 days

Figure 1. The sensory profile of the Feteasca neagra wines

Aromatic intensity
4
Quality tannins 3 Aromatic quality

Quantity tannins 1 Bouquet of pepper

0

The aroma of
Astringent
raisins

Dark chocolate Black blueberry

28 eyes/vine- 8 days 28 eyes/vine- 16 days

20 eyes/vine- 8 days 20 eyes/vine- 16 days
36 eyes/vine- 8 days 36 eyes/vine- 16 days

Figure 2. The sensory profile of the Cabernet Sauvignon wines

CONCLUSIONS
Extending the maceration-fermentation time from 8 to 16 days after the end of
alcoholic fermentation leads to important changes in the structure of red wines:
decreases the total anthocyanin content and the color intensity of the wine; increases
the content in total polyphenols, modifying the indices that characterize the phenolic
complex in wine.
319
 Red wines obtained by maceration-fermentation for 8 and 16 days recorded
increasing values of the anthocyanin content.
The extension of the maceration-fermentation period induces an increase in
the total polyphenols content

REFERENCES
Bourzeix M., 1975, Les anthocyanes du raisin et du vin, Vignes et Vins, nomero
special, p. 1-18
Bourzeix M., 1976, Les composes phenoliques des raisins et du vin. Leurs effets
sur la qualite. Rev. Frank. Oenol 63, 53-69
Kontek A., 1975, Evoluţia unor compuşi fenolici în decursul procesului de
maturare a strugurilor roşii, 1975, Analele Institutului de Cercetări pentru Viticultură şi
Vinificaţie Valea Călugărescă, Vol. VI, p. 499-509
Kontek Adriana, 1983, Date referitoare la conţinutul în compuşi fenolici al
strugurilor negri şi vinurilor roşii de Valea Călugărească, Anale ICVV., vol. X, pag.319-
325
Ribereau-Gayon, 1972.- Evolution des composes phenoliques en cours de la
maturation du raisins. Conn.Vigne Vin 6, p.161-175.
Ribéreau-Gayón, 1964 Les composés phénoliques du raisin et du vin.II. Les
flavonosides et les anthocyanosides. Ann. Physiol. Vég. 6:211-242
Tardea C. , 2007, Chimia si analiza vinului, Editura Ion Ionescu de la Brad, Iasi

320
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 Food produce process ing
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Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

THE BEHAVIOUR OF SOME GRAPEVINE GENOTYPES CREATED AT

INCDBH STEFANESTI ARGES, INITIAL MATERIAL G0, IN DEPOSITARY
GREENHOUSE CONDITIONS
1 1 1
Diana Elena Vizitiu , Gina Nedelea , Carmen Bejan , Emilia Vișoiu
1*
National Research and Development Institute for Biotechnology in Horticulture Ștefănești
Argeș, Romania
* Correspondence author. E-mail: [email protected]

Key words: starch, glucides, Auriu de Ştefăneşti, Perlette

ABSTRACT
This paper presents the behavior of the propagation Initial material belonging to
grapevine genotypes created at NRDIBH Stefanesti Arges. The obtained results after carrying
out the biochemical tests regarding the maturation of annual elements used in the multiplication
process of biological material shows a good ability of studied genotypes to adapt at the culture
conditions from the depositary greenhouse, fact due, largely, to the grapevine plasticity in the
situation of equilibrated supply with nutrients and water, as well as a good phytosanitary
protection throughout the entire vegetation period. In the case of studied genotypes the
accumulation of the reserve substances in woody tissue (soluble glucides and starch) was
positively correlated with the total length of shoots and with the matured chord portion.

INTRODUCTION
The grapevine culture occupies an important place in Romania and it would
not be possible without a high quality propagation material. In the strategy of viticultural
sector development, the Initial G0 vine propagation material is the first step in
multiplying the valuable genotypes for romanian viticulture, having in perspective a
particular importance in the grapevines genetic amelioration. Therefore, the depositary
with Initial G0 vine propagation material, from NRDIBH Stefanesti Arges must be
continuously enriched and studied, at the same time.
By maintaining the grapevine propagation Initial material (G0) in the depositary
greenhouse are fulfilled two actual aspects of Romanian viticulture: firstly it avoids the
natural or accidental disappearance of valuable autochthonous grapevines varieties
and, secondly, it protects and promotes an biological material with a very good status
by point of view sanitary and physiologically. Over the years, the NRDIBH Stefanesti
Arges was delivered to the vine research units G0 Initia propagation material as scion
and rootstock eye and rooted cuttings in pots in order to create the Base mother
plantation (Şerdinescu, 2011).

MATERIAL AND METHODS

The effectuated studies had as primary objective the appreciation of the
adaptation capacity of some grapevines genotypes (Auriu de Stefanesti, Perlette 10
St., Fetească alba 97 St., Muscat Ottonel 16 St., Merlot 202 St. and Burgund mare 86
St.) at the growing conditions from the depositary greenhouse by quantitative and
321
qualitative evaluation of woody biological material obtained annually during 5 years of
study, between 2009-2013.
Maturation degree evalution of strings was performed using biochemical tests
(for determining the wood content in water, soluble carbohydrates and starch), after the
period with the lowest temperatures (20.01.2009, 01.02.2010, 07.02.2011, 21.02.2012
01.31.2013). The analyzed sample was made up from internods fragments
(comprising nodes and internodes) from the entire length of string. The string
portions of the base, middle and top were analyzed separately, the final
result being the mean of three determinations. As working methods, were used the
standard ones, used in specialized laboratories. The free water content determination
from the woody material was assumed the drying of the material at 40ᵒC, up to
constant weight, and the determination of total water content it was carried out at
105ᵒC. The determination of soluble carbohidrates and starch from woody material
was effectuated by the colorimetric method, with anthrone reagent (Scott and Melvin,
1953).

RESULTS AND DISCUSSIONS

The wood maturation is a phisiological process influenced by environmental
factors (Burzo I. et al., 1999). During the periods when the ambient temperature
decrease at values between 10ᵒC and 0ᵒC it takes place the intensification of soluble
carbohydrates translocation process from leaves to perennial organs, and, their water
content gradually decreasing.
In the course of the study it was found that the plants from the depositary
greenhouse have completed all phenophases approximately with one month earlier in
all years studied (2009, 2010, 2011, 2012, 2013), compared to grapevines plants from
field.
The periodic registrations effectuated at the grapevine genotipes from
depositary greenhouse, showed that the conduct of the phenological phases was
closely correlated with the climatic variations specific to the greenhouse, so that, in the
study years, between 15.03 -04.04, took place: flowering 28.04-15.05, firstfruits 01.07-
31.07 full maturation: 20.07-15.08, strings maturation and leaf fall: 25.08-30.11; also
the studies have indicated, depending on the genotyp, a vegetation period of 185-250
days.
At the end of the vegetation period of each year, in depositary greenhouse, the
vegetative growths reached lengths of over 250 cm, from which a average of 204 cm
was maturated wood. The string diameter and the distance between the knots
constituted indicators in the quantitative assessment of the number of buds from the
maturated wood portion. Thus, the number of total buds was supperior for the studied
genotypes (210-250 eyes / 4 plants), where the distance between nodes was between
12-15 cm and the strings diameter 5-8 mm.
In this study, the temperature decrease in greenhouse occurred after
November 15, when it registered positive values of 3-4ᵒC in a longer period of time
(10-12 days). Thus, before leaf fall, took place the protein and carbohydrates
(poliglucide) hydrolysis from leaves and their translocation as amino acids,
respectively, soluble carbohydrates.

322
 Figure 1. The grapevine plants in the depositary greenhouse
The total water content obtained from the genotypes analyzed from the
depository greenhouse registered medium multiannual values between 47 and 50%
(Fig. 2d). The material derived from the clones for the production of white wines
(Feteasca alba 97 St. and Muscat Ottonel 16 St) registered an average content in total
water of 47.9 - 48.6% and those for the production of red wines (Merlot 202 St and
Burgund 86 St) it was 47.2 % (Fig. 2 b and c). At grapevine genotypes for table grapes
(Auriu de Stefanesti and Perlette) the multiannual averages were situated around 50%
(Fig. 2).

a b

c d
Figure 2. The total water content from the woody material obtained from grapevine
genotypes tested between 2009-2013 (a, b, c) and multiannual average (d)
In this study the bound water content from grapevine strings registred
appropriated multiannual average values (2.3 - 2.9%), observating its decrease at all
genotypes tested in 2010 and 2012 (Fig. 3 a, b, c). Quantitatively, the bound water
recorded higher values at Perlette 10 St. clone (fig. 3 a).

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 a b

c
Figure 3. The bound water content of woody material between 2009-2013 (a, b, c)
The glucides quantification showed a good maturation of the woody material at
all tested genotypes. At the table varieties, soluble glucides content was situated
between 10.3% dry substance at Auriu de Stefanesti and 12.7% at Perlette 10 St.clone
(multiannual average - Fig. 4 d). In the viticultural year 2012-2013 both genotypes have
accumulated significant amounts of glucides in wood (12.3% at Auriu de Stefanesti
varieties; 13.9% at Perlette 10 St. clone). At the grapevine genotypes intended for
quality red and white wines, the soluble glucides acumulation were between 10.1 % -
Burgund 86 St. and 11.8 % - Muscat Ottonel 16 St., accepted limits for a biological
material found in the rest period, corresponding to December and January, when the
soluble glucides content reaches a maximum of 10-12% glucides (Fig. 4 a, b and c).

a b

c d
Figure 4. The soluble glucides content between 2009-2013 (a, b, c) and the
multiannual average of soluble glucides and starch (d)
Regarding the starch content from canes, this recorded values close to the
normal in January and February, framing between 3.8 - 6.1% (multiannual average).
Higher starch accumulation were reported also in the viticultural year 2012-2013 at
324
clones for quality red wines (Merlot 202 Şt. - 7.3 %, Burgund 86 Şt. - 7.4 %). The
Perlette 10 St clone registred minimal value of starch in all studied years (Fig. 5 a, b
and c).

a b

c
Figure 5. The starch content of woody between 2009-2013 (a, b, c)
The studied genotypes have accumulated in the maturity stage of wood the
important quantities of sugars due to the optimal nutritional and health status of hubs,
but also due to the mild conditions from greenhouse where the minimum temperature
recorded values up to -10oC, when, in the field conditions, absolute minimum
temperature of air was -22oC (last decade of January 2010). The content evolution of
the total glucides from woody material belonging the clones and the varieties studied
between 2009-2013 is shown in Figure 6 a, b and c.

a b

c d
Figure 6. The total glucides content of woody material between 2009-2013 (a,
b, c) and the multiannual average (d)
325
 The multiannual average indicated accumulation of total sugars between 14.7
- 17.6%, which led at obtaining of superior primary buds viability, namely 97.2 - 99.6%
(Fig. 6 d). Meanwhile, the buds viability at annual woody elements harvested from the
studied genotypes was positively correlated with accumulated amount of total glucides.
Similar results for the obtaining of the quality viticultural planting material in
protected spaces has been achieved at Feteasca neagra 6 St., Pinot noir 3 St.,
Cabernet Sauvignon 131 St., Fetească regală 72 St. (Bejan şi colab, 2013) grapevine
clones.
The results obtained after biochemical determinations on the biological
material, reveals that the soluble glucides and starch acumulation in woody tissue was
correlated with total length of shoots and with the matured chord portion.

CONCLUSIONS
At the studied genotypes the contents of the woody material in bound water
was recorded multiannual average values between 2.3 - 2.9%, observing a decrease
of this indes at all tested genotypes between 2010 – 2012. quantitatively bound water
showed higher values at Perlette 10 St.The total water content of material obtained
from the analyzed genotypes recorded multiannual average values between 47 and
50%, values characteristic of a mature wood. In 2012-2013 years were recorded
significant amounts of soluble glucides at the table grapes genotypes and higher starch
accumulation at varieties clones for quality red wines.
The acumulation of soluble glucides and starch was fluctuated depending on
the genotype and on the specific climate of the year in which the evaluation was
conducted, but was within the limits of a biological material found during the rest
period.
The obtained results show a very good capacity for adaptation of studied
genotypes at depositary greenhouse culture conditions, due the grapevine plasticity in
the conditions of a equilibrated supply with nutrients and water, as well as a good
phytosanitary protection throughout the entire vegetation period.

REFERENCES
Şerdinescu A., 2011, Sprijinul cercetării, la dezvoltarea producţiei de material
săditor, Info AMSEM. Seminţe şi material săditor. Anul XIII, Numărul 4, pp. 18-20,
ISSN 2068-6862.
Burzo I., Toma S., Olteanu I., Dejeu L., Delian E., Hoza D., 1999, Fiziologia
plantelor de cultura, Inteprinderea Editorial-Poligrafica Stiinta Chisinau, Vol.3, pp 440.
Scott T.A. şi Melvin E.H., Determination of dextran with anthrone, 1953, Anal.
Chem. 25:1656–1661.
Bejan Carmen, Nedelea Gina, Visoiu Emilia, Biochemical evaluation of the
quality of G0 grapevine initial planting material, 2013, The 36rd World Congress of vine
and wine, Vine and Wine between Tradition and Modernity, Book of abstracts, pp. 195-
196.

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 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
 Food produce process ing
UNIVERSITY OF CRAIO VA
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

THE LABEL. THE EUROPEAN WINE ”PASSPORT”

1 1* 1 1
Voinea-Mic Cătălin , Giugea Nicolae , Berta Corina , Mărăcineanu Liviu
1*
University of Craiova, Faculty of Horticulture
* Correspondence author. E-mail: [email protected]

Key words: label, geographical indications, controlled designation of origin, specific traditional
mentions, consumer
ABSTRACT
Starting from the European desideratum to protect the legitimate interests of wine
consumers as important as the producers' interest in capitalizing their production depending on
the quality of the supplied products, the European Community has established higher standards
than the national legislation, designed to regulate how to present the wine once it is
commercialized.

INTRODUCTION
The label is a genuine wine identity document, a passport in international trade
relations, along with the fiscal and commercial documents accompanying the product,
the true business card of the bottled product. By referring strictly to the common
clients, the label is, in fact, the major milestone in the wine selection process, starting
from the chromatic aspect and ending with the strict specialty inscriptions on it.

MATERIAL AND METHODS

This paper was based on the European regulations and European
jurisprudence in the field, using the classical methods of legal interpretation of
legislation and legal analysis on the text of parts of the procedural documents in some
cases solved by the CJEU.

RESULTS AND DISCUSSIONS

According to European standards (Notice Regulation no. 1493/1999 on the
common organization of the wine market, Annex no. VII), the label is defined as "a set
of descriptions and other mentions, insignia, illustrations or trademarks that
characterize the product, placed on the same container, including on the closure
devices, or on the label attached to the container".
From the legal definition, several conclusions can be drawn:
- the label may contain illustrations, insignia or distinguishing marks -
trademarks;
- the notion of "label" also includes closing devices, as far as they are fit to be
inscribed, illustrated;
- in addition to the classic attachment formula or sticking on the container, the
label may also be attached to the container (usually with a string, ribbon, etc.); there
should be mentioned that the packaging of the product does not represent the labeling;
the packaging is done with paper, straws of any kind, cardboard, cassettes, etc. and it
327
is intended exclusively for the transport of one or more containers and/or their
presentation for sale to the final consumer.
Beyond the manufacturer's desire for product presentation through the label,
the European standards establish that, from the date of putting into circulation, any
container with a nominal capacity of not more than 60 liters must be labeled. It is
obvious that each wine producer is tempted to produce a label with a maximum impact
on consumers. The right to ownership of the wine, of the container, and implicitly of the
label, is not equivalent to the trader's absolute freedom to insert the specifications on
the product presentation label. The consumer protection and, implicitly, the protection
of the interests of traders of similar products require that the description and
presentation of the products through the presentation label are not misleading or likely
to cause confusion to consumers. The limits of the right to present the wine product are
set at European level by Regulation no. 1493/1999 (In force since 14.07.1999,
published in JOCE no. 179 of 14.07.1999) on the common organization of the wine
market, an European act containing rules obliging the insertion of certain mentions,
authorization rules for certain mentions, the protection and control of mentions, the use
of geographical indications and traditional mentions, rules on imported product labeling
(to the extent that the regulation permits these operations), on the authorization of the
use of consumer information, etc. The label must necessarily accompany the wine
categories specifically specified by the Frame Regulation, the content of which is
formed of mandatory and optional mentions. Thus, in the categories of mandatory
mentions referring to wine categories and sales denominations, we distinguish:
a) labeling of table wines with a geographical indication and wines with a
controlled designation of origin (DOC); the label will contain the name of the bottling
company and the name of the locality and the Member State of which it is part; in the
case of bottling (Operation of the product introduction in a container of less than 60
liters for the purpose of sale) in containers of more than 60 liters, it will be compulsory
to includ the name of the sender;
b) the labeling of wines coming from third party countries other than liqueur
wines, petillant wine, whether or not sparkling, as listed in Annex I to the framework
Regulation; the name of the importer will be placed on the label and if the imported
wine has been bottled in the EU, the name of the bottler will be also included;
c) the labeling of liqueur wines, sparkling or non-sparkling petillant wines as
referred to in Annex I to the framework Regulation, including those of third party
country origin; in the case of these wines, the mention of the sales name of the
product, the nominal volume, the alcoholic strength gained in volume and the lot
number must be included (According to Council Directive EEC 89/396 of 14 July 1989
on indications or marks identifying the lot to which a food product belongs);
d) the sales name, which is composed of the following terms, depending on
the characteristics of the product, as follows:
i. for table wines - the mention "table wine" accompanied by the name of the
EU country where the grapes were produced and vinified when the export is made to
another Member State; the mention "table wine" accompanied by the phrase "blend of
wines from different European Community countries" when wines are the result of
grapes processing from several EU countries; the mention 'table wine' accompanied by
the words 'wine obtained in ... from grapes harvested from ...' when the wines are
harvested in a Member State and are being vinified in another Member State;
ii. for table wines with geographical indication - the mention "table wine"
followed by the name of the geographical unit; in the case when there have been
established known names in order to determine the specific geographical indication of

328
a country at European level (eg "regional wine"), the reference "table wine" is no longer
mandatory, and the geographical area followed by this mention is inserted;
iii, for wines with a controlled designation of origin (VDOC), the term "VDOC"
(or "wine with a controlled designation of origin"), the mention "LWDOC." (or "liqueur
wine with a controlled designation of origin"), the mention "PWDOC" (or "petillant wine
with a controlled designation of origin"), a specific traditional mention, as we will
discuss below;
v. for imported wines - the word "wine" accompanied by the name of the
country of origin and the name of the relevant geographical area (if applicable);
vi. for liqueur wines - the term "liqueur wine";
vii. for petillant wines - the mention "petillant wine";
viii. for sparkling petillant wines - the term "sparkling petillant wine";
The optional mentions, called by the European legislator "optional indications",
allow manufacturers /traders to add on the label the following indications designed to
better characterize the product, namely: the name, address and function of one of the
persons who participated in the trading, type of product, specific colour, harvest year,
name of one or more vine varieties, a prize, a medal or a contest (Notice the comment
below on the medals and distinctions inserted on the labels), indications on the method
of obtaining and production, the name of the winemaking company, additional
traditional mentions, etc.
As part of the optional indications, the European legislator has also inserted
additional traditional mentions which, along with other information, can provide a clear
picture of the product. Thus, the mandatory mentions on the label, supplemented with
regulated optional indications, may be supplemented with other indications, provided
that the additional text does not create a risk of confusion among addressees as
regards mandatory mentions and optional indications. According to art. 23 of
Regulation no. 753/2002 laying down certain detailed rules for the application of
Council Regulation (EC) No. 1493/1999 as regards the designation, nomination,
presentation and protection of certain wine products, through the words "Additional
traditional mention" shall be used to indicate the method of production, preparation,
maturation or the quality, colour, type of place or a special event related to the history
of the wine in question. Both theoretically and practically, this comprehensive definition
grants the Member States the right to designate the nationally produced wines that
meet the criteria mentioned in the definition established at European level.
If, in the case of mandatory and optional mentions expressly regulated, the
things cannot be interpreted, the problems arise in the case of optional mentions that
may be subject to violations of rights already authorized. In this respect, the European
legislation restricts the use of additional information other than the mandatory ones,
establishing as a barrier the situation in which there may be a risk of confusion among
consumers regarding the regulated mandatory and optional mentions.
It should be noted that this risk of confusion must be assessed by an average
consumer, not addressed to a wine specialist. The CJEU has stated that "... it must be
demonstrated that the use of the mark is indeed likely to mislead the targeted
consumers and thereby influence their economic behavior. In this regard, the national
court must take into account the presumed expectations regarding these indications of
an average consumer, normally informed, sufficiently attentive and competent. "
(Regulation of 28 January 1999, Sektkellerei Kessler (C-303/97, ECR I-513, paragraph
32) (risk of confusion / labeling of German sparkling wine - Sekt)
In order to avoid confusion with other similar products, the European
legislation requires that on the label applied to the container the following elements
should be necessarily inserted: the identity and quality of natural and legal persons or
329
associates who have been or are involved in the production or commercial trade of
wine, composition, alcohol level by volume, colour, origin or provenance, quality, vine
varieties, harvest year or nominal volume of the containers. The mandatory indications
on the description, designation, presentation and protection of certain wine products
(Except for sparkling wines) must be presented in a clustered, clearly visible, clear and
distinct manner from the other inscriptions on the label. The importer mentions and
batch number may be inserted separately from the other mandatory information. The
ink or printing method must be resistant to external factors, so as to avoid the risk of
deleting this information during the transport, handling process etc.
Regarding the alcoholic strength by volume, this mention must be given on
labels with a height of at least 5 millimeters of characters if the nominal volume is more
than 100 centilitres, of at least 3 millimeters if the volume is lower or equal to 100
centilitres and bigger than 20 centilitres and at least 2 millimeters if the volume is less
than or equal to 20 centilitres. The obtained distinctions or medals can be mentioned
on the label of the table wines with geographical indication or wines QWPSR only if
these distinctions have been awarded to the batch of wine in question in competitions
authorized by EU Member States or third party States to the extent that they were
organized under advertising conditions designed to remove any suspicion of doubt.
Thus, the mention of "wine awarded at ..." inserted on a label applied to a batch of
wine which differs from the legal batch equates to an infringement of European rules
on consumer protection and market competitors.
Regarding the language used for the mentions on the label, the European
legislation requires to be one or more of the official languages of the European
Community, so that the information of the consumer to be easier understood. Given
their specificity, the mentions referring to the name of the region of production/name of
another geographical unit, the traditional mentions and additional traditional mentions
(By "additional traditional mention", it is indicated the method of production,
elaboration, maturation or quality, colour, type of place or a special event linked to the
history of the wine in question), the name of the winemaking company, the mentions
about bottling can be expressed in the official language(s) of the member state territory
on which that product was produced. Also in the countries where other languages are
spoken besides the official language, these specific mentions may also be expressed
in the language traditionally used in that area if it is closely related to that product.

CONCLUSIONS
The label represents, together with other accompanying fiscal and customs
documents, the way in which wine entries into consumer consciousness, contributing
greatly to building quality standards both in terms of protecting the final recipient and in
establishing an economic order based on the principles which governs loyal
competition between manufacturers. Editing legal requirements requiring mandatory
mentions insertions on the labels attached to bottled products does not constitute an
infringement of the right to promote, but double guarantees in favor of both consumers
and honest traders in the vast field of wine production and trading.

REFERENCES
*** 1999, Regulamentului (CE) nr. 1493 privind organizarea comuna a pietei
viticole, in vigoare din 4 mai 2002, publicat in JOCE nr. 118 din 4 mai 2002;
*** 2002, Regulamentul nr. 753 de stabilire a anumitor norme de aplicare a
Regulamentului (CE) nr. 1493/1999

330
 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
 Food produce process ing
UNIVERSITY OF CRAIO VA
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

WINE DESCRIPTION. THE ADDITIONAL TRADITIONAL MENTION

1 1* 1 1
Voinea-Mic Cătălin , Giugea Nicolae , Berta Corina , Mărăcineanu Liviu
1*
University of Craiova, Faculty of Horticulture
* Correspondence author. E-mail: [email protected]

Key words: additional traditional mention, labels, optional mentions, misleading, average
consumer

ABSTRACT
The description of wine products by means of additional traditional mentions is not
limited to the regulations expressly provided by the Community or national legislator, and it can
take various forms of expression as long as there is no concrete misleading of the consumer.

INTRODUCTION
Given the very wide appreciation left by the European legislator for additional
traditional mentions, referring to the established principle of legislation liberalizing on
product names marketed in the European Union, this article aims to analyze the extent
to which the trading of wine products under strictly unregulated denominations by prior
authorization, is permitted. More specifically, it is interested in the extent to which
additional traditional mentions may be inserted on labels insofar as they are not part of
the mandatory and optional mentions expressly regulated by the legislator.

MATERIAL AND METHODS

This article was based on European regulations and European jurisprudence
in the field, using the classical methods of legal interpretation of legislation and legal
analysis of the text of parts of the procedural documents in some cases solved by the
CJEU.

RESULTS AND DISCUSSIONS

According to art. 23 of Regulation no. 753/2002 laying down certain detailed
rules for the application of Council Regulation (EC) No 1493/1999 (Concerning the
common organization of the wine market, in force since 4 May 2002, published in
JOCE no. 118 of May 4, 2002) (hereinafter referred to as the "Implementing
Regulation"), the term "Additional traditional mention" means the term used to indicate
the method of production, preparation, maturation or quality, colour, type of place or
special event related to the history of the wine in question. This comprehensive
definition leaves the Member States the option of designating the wines produced at
national level on the basis of the criteria set out in the definition established at
European level.
Given the very wide margin of appreciation left by the European legislator
regarding the additional traditional mentions, referring to the established principle of
the legislation liberalization of denomination, this article aims to analyze to what extent
331
the trading of wine products under unregulated denominations strictly subject to prior
authorization is permitted. More specifically, it is interested in the extent to which
additional traditional mentions can be inserted on labels insofar as they are not part of
the mandatory and optional mentions expressly regulated by the legislator.
If the range of these mentions is quite wide, given the very permissive
definition, we make it clear that there are regulations that expressly refer to the
necessity and / or the possibility to insert the additional traditional mention.
Thus, as an example, we introduced the provisions of art. 14 par. (1) lit. (c) of
the "Implementing Regulation" for grape must in fermentation intended for direct
human consumption or for wines obtained in the Community from over-maturated
grapes, where it is expressly stated that the name under which these products may be
put up for sale may indicate, besides the name of the geographical unit of production
and the product category (Grape must in fermentation" or "wines obtained from over-
maturated grapes") which are mandatory, also a specific traditional mention, to the
extent that it does not become repetitive to the name under which the product is
marketed.
Besides the punctual regulations, the limits of the use of these mentions, or
rather the condition that limits the use of this type of description, is given by art. 6 par.
(1) of the "Implementing Regulation" according to which the labeling of the concerned
products may be "supplemented with other indications, provided that the additional text
does not create a risk of confusion for the persons to whom it is addressed", in
particular regarding the mandatory and optional mentions.
It should be noted that, for an effective analysis of the permissiveness of an
additional traditional mention, we must refer, on the one hand, to the list of mandatory
mentions and optional mentions stipulated in section A.1. and section B.1. of Annex VII
to Regulation (EC) No. No 1493/1999 on the common organization of the wine market
and on the other hand to the risk of confusion, misleading, which this mention may
bring to consumers.
If, regarding the mandatory mentions, the situation is clear in the sense that an
additional traditional mention does not have to create confusion with a mandatory
mention, problems arise when referring to an optional mention, regulated by the
legislator. More specifically, in practice, the question of the lawfulness of the
presentation of a wine product with an additional traditional mention which does not
appear to be regulated expressis verbis by the legislator has been raised.
Starting from the regulation of mandatory and optional indications, we can say
that any indication referring to the method of production, maturation elaboration and
wine quality can be authorized not only as an optional indication regulated under point
B.1. (b) of the fifth indent of Annex VII to Regulation No. 1493/1999 because the
labeling of the products within the community can be supplemented with additional
traditional mentions, according to the rules provided by the producer state. In this
respect, Art. 23 of the "Implementing Regulation" provides that Member States shall
lay down the framework and conditions for the use of indications concerning the
method of production or the methods for the preparation of table wines with a
geographical indication or of wines QWPSR (Quality wines produced in specified
regions) for the wines produced on their territory.
In support of the conclusion that traders may use additional traditional
mentions, unregulated, the European rules provided in point B.3 of Annex VII to
Regulation No. 1493/1999 that, in the case of table wines, table wines with a
geographical indication, wines with a controlled designation of origin (VDOC), liqueur
wines, petillant wines and sparkling petillant wines (Mentioned in Annex I of the
Regulation no. 1493/1999 on the common organization of the wine market, in force
332
since 14.07.1999, published in JOCE no. 179 of 14.07.1999), including those coming
from third party countries of the European Union, the labeling may contain other
information than the mandatory and optional mentions expressly referred to in the
European regulation framework.
Basically, from the point of view of the regulation, we consider that the only
obstacle to the use of an additional traditional mention, other than those expressly
regulated, is the existence of an express provision prohibiting the use of that mention
at Community or national level. In other words, authorizing the use of an additional
traditional mention should not be inflexible in relation to the existence or not of the
regulation but must be analyzed on a case-by-case basis whether or not there is a risk
of misleading the consumers.
Thus, the second condition to be met with regard to specific traditional
mentions, is that there is no risk of misleading the consumer. In other words, each
case must be analyzed in concrete terms to determine whether a particular name is
misleading or not. "The purely abstract danger of creating the appearance that it is a
protected designation cannot lead to the prohibition of the use of an unregulated
indication, as long as there is no actual misleading of the consumer"(Conclusions of
the Advocate General VERIKA TRSTENJAK delivered on 25 October 2007 in Case C-
285/06 Heinrich Stefan Schneider vs Land Rheinland - Pfalz, published at eur-
lex.europa.eu on 25 October 2007).
Regarding the consumer, or the danger of misleading the consumer, we notice
that always this risk of confusion must be appreciated by an average consumer, not
reported to a wine specialist. The CJEU (Court of Justice of the European Union) has
stated that "... it must be demonstrated that the use of the mark is indeed likely to
mislead the target consumers and thereby influence their economic behavior. In this
regard, the national court must take into account the presumed expectations regarding
these indications of an average consumer, normally informed, sufficiently attentive and
competent” (Resolution on 28 January 1999, Sektkellerei Kessler in Case C-303/97
ECR, p. I-513, paragraph 32; (risk of confusion / labeling of German sparkling wine -
'Sekt').
It is well-known that this consumer will not know the European regulations
regarding names, traditional or other additional regulated mentions, the appreciation of
the risk of confusion being reported by the ability of the unregulated mention to mislead
this consumer always by reference to mandatory or optional mentions already existing
in the market. Always an increased attention in the assessment of the risk of
misleading should be based on the stipulation of art. 24 paragraph (2) of the
"Implementation Regulation", according to which an existing traditional mention is
protected against "any illicit, imitation or use of citations, even if the mention in
question is accompanied by an expression as follows <<kind>>, <<type>>,
<<imitation>>, <<brand>> or other similar terms”.
In this regard, as an example, we demonstrate that there is no imitation or
citation unless the language used is the same as that in which the already protected
traditional mentions are indicated. Thus, the legislator limited the scope of protecting
traditional mentions to their own language. The use of the German expression
"Reserve" for German wines as a specific traditional mention contravenes Annex no. III
to the "Implementing Regulation" which recorded this mention for Austrian wines and
which would mislead the consumers of the Austrian wines holding this mention.
However, derivative mentions such as "Privat-Reserve" may be used instead, without
contravening the European standards, the legislator even allowing translations of the
mentions, but without allowing the copying of already used and legally protected
names (it is forbidden to have the already used names in the language of origin).
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 CONCLUSIONS
In conclusion we can specify that specific traditional mentions can be used
without barriers as long as they do not contravene the European or national norms
defining these categories of mentions, the legality of their use being censored on a
case by case basis in terms of avoiding the mislead of the consumer, in relation to
already existing and protected names at Community level.

REFERENCES
*** 1999, Curtea de Justitie a Uniunii Europene, Hotararea din 28 ianuarie 1999,
Sektkellerei Kessler C-303/97, Rec., p. I-513, pct. 32;
*** 1999, Regulamentului (CE) nr. 1493/1999 privind organizarea comuna a pietei
viticole, in vigoare din 4 mai 2002, publicat in JOCE nr. 118 din 4 mai 2002;
*** 2002, Regulamentul nr. 753/2002 de stabilire a anumitor norme de aplicare a
Regulamentului (CE) nr. 1493/1999;
*** 2007, Concluziile Avocatului General VERIKA TRSTENJAK prezentate la 25
octombrie 2007 in Cauza C-285/06 Heinrich Stefan Schneider Impotriva Land
Rheinland – Pfalz, publicate in eur-lex.europa.eu din 25 octombrie 2007.

334
 Seri es:  Bio logy
ANNALS OF T HE  Horticu lture
 Food produce process ing
UNIVERSITY OF CRAIO VA
techno lo gy
Vol. XXII (LVI II ) - 2017  Environ menta l e ng ineer ing

EFFECT OF RESIDUE GREEN ROBUSTA COFFEE, OBTAINED AFTER

THE REMOVAL OF HOT AQUEOUS EXTRACT, ON SOIL CHEMICAL
PROPERTIES
1 1 1 1
Wogiatzi Eleni , Gougoulias Nikolaos *, Giannoulis Kyriakos , Miliakidi Eleni-Maria
1
Department of Agricultural Technology, Technological Educational Institute of Thessaly, 41110 Larissa,
Greece
*Correspondence author. email: [email protected]

Key words: Green coffee; Soil organic matter biodegradation; Waste

ABSTRACT
The effects of the green coffee residue, obtained after from the removal of the extract
with hot water, on soil chemical properties for an incubation experiment, were studied. The air
dried residue incorporated at four different rates (0, 0.3, 0.6, and 0.9g per 50g of soil mixture with
9,84g of manure) resulted in increase in organic carbon mineralization, at the two higher rates
by 17.4 and 24.5%, respectively, at compared to the control. The addition of the residue Robusta
in all treatments increased available forms of Mn, Fe and organic P, while at the two highest
rates increased the K-exchangeable and N-NO3-, and at the highest dose increased the N-NH4+,
and available Cu. Available forms of P and Zn was not significantly affected by the residue
addition. The results of this study indicated that residue coffee could be applied to the soil
without any extremely negative effect on the soil chemical properties.

INTRODUCTION
In biological agriculture is mandatory to replace chemicals with natural
additives. Between them, various organic residues for their role in the soil fertility, the
plant essential oils for their role as pesticides against on the soil pathogens
(Gougoulias et al. 2010, Gravanis et al. 2004, Hartemink and Hutting 2005).
Green coffee beans contain a tremendous number of phenolic compounds,
such as chlorogenic acids, ferulic acid, tannins, proanthocyanidins, lignans, and
tocopherols, which exert strong antioxidant effect (Borelli et al. 2002, Farah and
Donangelo 2006).
Literature on coffee waste as a soil amendment is limited, however studies
have shown that coffee waste is a valuable organic fertilizer, particularly for highly poor
soils (Zake et al. 2000). In addition, coffee waste can be used as a fertilizer for sandy
soils and an improvement of the soil physico-chemical properties (Kasongo et al.
2011).
The objectives of this research were to study, in the laboratory, the effect of the
application of the green coffee solid residue obtained after the removal of the extract
with hot water on soil organic matter and on soil chemical properties.

335
 MATERIAL AND METHODS
Commercial green Robusta coffee beans from the local market in Larisa were
dried in a dark place at room temperature, finely ground, shaken at 150 rpm for 30 min
using boiling water. The extraction of the green Robusta coffee with boiling water
solvent results in the extraction of phenol compounds equal to16.86 mg (GAE)/g DW
and antiradical activity (DPPH•) 93.6 (μmol Trolox /g) DW. Green Robusta coffee
beans are a source of bioactive components and the extracts can find applications as
food additives, either in the form of powder or as extracts highly concentrated at
polyphenols and antioxidant activity. The green coffee solid residue, which remained
after the removal of the extract with hot water, was air dried and was used for the
incubation experiment.
Incubation experiment: In this study, 9.84 g of a sheep manure containing
2.5 g of organic matter (Table 1), obtained from the farming establishments of TEI of
Thessaly, and was added to 50 g of air-dried soil that was poor in organic matter,
derived from the same region (Table 1). Into 50 g of this soil plus 9.84 g of manure, 0,
0.3, 0.6 and 0.9 g of air-dried and well milled green Robusta coffee residue, were
added. Thus an experimental unit is constituted by 50 g of soil, 9.84 g of manure and a
variable amount of residue green Robusta coffee. In the incubator, the treatments were
prepared in four replicates and kept at 28 0C for a period of 15 weeks. During the first
three weeks of the incubation period, the moisture was maintained at two-thirds of field
capacity, but for the next three weeks the soils were left to dry. This process was
repeated until the end of the incubation period according to (Wu and Brookes 2005)
they reported that the alternation of drying and rewetting soil samples enhances
mineralization of both soil biomass organic matter and non-biomass organic matter.
Soil aeration and regulation of moisture enhance the growth and metabolic activity of
aerobic soil microorganism (Gordon et al. 2008). At the end of the incubation period,
soil samples were analyzed.
Methods of analyses: Samples were analyzed using the following methods
which are referred by (Page et al. 1982).
Organic matter was analyzed by chemical oxidation with 1 mol/l K2Cr2O7 and
titration of the remaining reagent with 0.5 mol/l FeSO4.
Both ammonium and nitrate nitrogen were extracted with 0.5 mol/l CaCl2 and
estimated by distillation in the presence of MgO and Devarda's alloy, respectively.
Available P forms (Olsen P) was extracted with 0.5 mol/l NaHCO3 and measured by
spectroscopy. Exchangeable forms of potassium and sodium ware extracted with 1
mol/l CH3COONH4 and measured by flame photometer.
Organic phosphorus was measured after mineralization by combustion of the
sample and subtraction of the mineral phosphorus amounts, which had previously
been estimated in the laboratory. The mineral amounts were extracted with 1 mol/l
H2SO4 and all forms were measured by spectroscopy.
Available forms of Mn, Zn, and Cu were extracted with DTPA (diethylene
triamine pentaacetic acid 0.005 mol/l + CaCl2 0.01 mol/l + triethanolamine 0.1 mol/l)
and measured by atomic absorption.
For the determination of total metals Mn, Cu and Zn , 1 g of material,
digestion at 3500C + 10 ml HNO3 + 5 ml HCLO4. According to the method described by
(Varian 1989), the samples were analyzed by Atomic Absorption (Spectroscopy Varian
Spectra AA 10 plus), with the use of flame and air-acetylene mixture.
Statistical analysis: Data analysis was made using the MINITAB (Ryan et al.
2005) statistical package. Analysis of variance was used to assess treatments effect.
Mean separation was made using Tukey's test when significant differences (P=0.05)
between treatments were found.
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 Table 1
Chemical properties of soil samples, manure and residue green Robusta coffee used
in the experiment

Property Soil *Manure Residue green

Robusta coffee
Texture Sandy Loam
pH 7.94 ± 0.4 8.85 ± 0.6
EC, dS/m 0.46 ± 0.05 2.33 ± 0.3
Organic matter (%) 0.85 ± 0.05 25.41 ± 1.3
CaCO3 (%) 8.68 ± 0.6 7.95 ± 0.6
N -Total (g/kg) 1.26 ± 0.10 9.36 ± 0.6 20.79 ± 0.9
+
N-NH4 (mg/kg) 47.2 ± 9.3
N-NO3- (mg/kg) 113.2 ± 14.2
K-exchangeable (mg/kg) 223.7 ± 7.6
K-Total (g/kg) 4.73 ± 0.24 5.30 ± 0.5 0.43 ±0.02
Na-exchangeable (mg/kg) 222.3 ±9.6
Na-Total (g/kg) 0.68 ± 0.04 0.47 ± 0.02 0.17 ±0.01
CEC (cmol/kg) 19.8 ± 1.2
P -Olsen (mg/kg) 14.2 ± 3.3
P -Total (g/kg) 0.32 ± 0.04 4.37 ± 0.2 1.51± 0.09
Cu –DTPA (mg/kg) 0.83 ± 0.05
Zn -DTPA (mg/kg) 1.31 ± 0.09
Mn -DTPA (mg/kg) 2.33 ± 0.13
Cu –Total (mg/kg) 27.5 ± 2.7 3.97 ± 0.22
Zn -Total (mg/kg) 271.4 ± 15.2 0.12 ± 0.01
Mn -Total (mg/kg) 643.5 ± 36.4 2.02 ± 0.14
Fe-Total (g/kg) 8.14 ± 0.3 0.099 ± 0.002
Mg-Total (g/kg) 14.58 ± 0.9 1.35 ± 0.08
*Digested sheep manure four months; Electrical conductivity, (EC) and soil pH is
determined in (1:5) soil/water extract; Data represent average means and SE
deviation. (n)=4.

RESULTS AND DISCUSSIONS

Decomposition of organic matter was increased with the increase of residue
Robusta concentration in soil, as compared with the control (soil plus manure). In
particular, the decomposition of organic matter in soil where the two higher rates of the
residue Robusta were incorporated were 17.4 and 24.5 % higher, respectively,
compared to the control (Table 2). However, was not significantly affected by the
smaller quantity addition of the residue Robusta.
The analysis of variance performed for the electrical conductivity data
indicated that the electrical conductivity was slightly increased by the added of the
residue Robusta. In particular, the highest residue Robusta concentration, increased
electrical conductivity by 24.5%, without any risk of salinity (Table 2). In contrast, the
soil pH was reduced by the residue Robusta addition in all treatments at comparison
with control (soil plus manure).

337
 Ammonium was increased by the addition of the highest rate of the residue
Robusta and was not significantly affected by the two lower quantities of the residue
Robusta. Also, nitrate nitrogen was increased by the addition of the residue Robusta
of the two upper rates, and it was not significantly affected by the smaller quantity of
the residue Robusta (Table 2). In particular, the addition of residue Robusta at the
highest dose increased ammonium and nitrate nitrogen by 202% and 43.5%
respectively in comparison with the control (soil plus manure).
The analysis of variance performed for the organic phosphorus and available P
data indicated that the organic phosphorus was increased by the residue Robusta
added. The greatest increase was caused by the greatest dose of residue Robusta
(Table 2). However, available P was not significantly affected by the residue Robusta
addition.

Table 2
Chemical properties of soil mixtures at the end of the incubation period

Properties Treatments
C R1 R2 R3
C-organic mineralization 257.4b 238.0b 302.1a 320.4a
(g/kg initial organic carbon)
EC (dS/m) 1.06b 1.12b 1.15b 1.32a
pH 7.01a 6.88b 6.87b 6.86b
+
N-NH4 (mg/kg) 44.0c 54.7c 47.25c 133.0a
N-NO3- (mg/kg) 543.2c 528.35c 654.5b 779.8a
P-Olsen (mg/kg) 216.8a 212.3a 207.8a 202.1a
P-Organic (mg/kg) 620c 749.5b 1507.3a 1333.2a
K- exchangeable (mg/kg) 613.1c 624.1c 690.43b 817.46a
Na- exchangeable (meq/100 g) 0.511a 0.474a 0.486a 0.524a
CEC (mmol/kg ) 16.6c 16c 17.1b 18.9a
Cu-DTPA (mg/kg) 0.87b 0.68b 0.87b 1.26a
Zn-DTPA (mg/kg) 6.36a 6.30a 6.40a 6.80a
Mn-DTPA (mg/kg) 2.10c 2.76b 3.32a 3.43a
Fe-DTPA (mg/kg) 0.48b 1.09a 1.38a 1.19a
Electrical conductivity, (EC) and pH measured in (1:5) soil/water extract. For each
chemical property of soil mixtures, lines of table with the same letter do not differ
significantly according to the Tukey‟s test (P=0.05). C, control (50g soil plus 9.84g
manure); R1, R2 or R3, control and residue Robusta 0.3, 0.6 or 0.9g, respectively.

The exchangeable form of potassium was significantly affected at the end of

the incubation period by the residue Robusta addition. In particular, the exchangeable
form of potassium was increased by the two highest rates of the residue Robusta by
12.6% and 33.3% respectively compared to the control. However, exchangeable Na
was not significantly affected by the residue Robusta addition (Table 2).
The available copper was increased by the addition of the highest rate of the
residue Robusta and was not significantly affected by the two lower quantities of the
residue Robusta. The addition of the residue Robusta at the highest dose increased
available copper by 43.8% at comparison to the control. However, available zinc was
338
not significantly affected by the residue Robusta addition. The available manganese
was increased with increasing residues Robusta concentration by 31.4, 58.1 and
63.3% respectively compared to the control. Also, available iron was increased by the
residue Robusta addition in all treatments at comparison to the control by 127.1% to
187.5%.
The total forms of potassium and phosphorus was increased at the end of the
incubation period by the residue Robusta addition at comparison to the control. In
particular, total potassium increased of the highest rate addition of residue Robusta by
6.3% compared to the control, while total phosphorus was increased by the two
highest rates of the residue Robusta by 43.9% and 44.5% respectively, at comparison
to the control (soil plus manure). However, total forms of sodium, magnesium, copper,
zinc, manganese and iron was not significantly affected by the residue Robusta
addition at comparison to the control.

Table 3
Total forms of elements of the soil mixtures at the end of the incubation period

Properties Treatments
C R1 R2 R3
Na-total (g/kg) 0.41a 0.40a 0.45a 0.41a
K-total (g/kg) 6.23b 6.04b 6.23b 6.62a
Mg-total (g/kg) 6.46a 6.38a 6.27a 6.28a
P-Total (g/kg) 1.82b 1.88b 2.62a 2.63b
Fe-total (g/kg) 22.47a 21.69a 21.89a 21.98a
Cu-total (mg/kg) 24.15a 19.32a 19.32a 19.32a
Zn-total (mg/kg) 84.22a 94.28a 84.22a 88.92a
Mn-total (mg/kg) 828.7a 788.2a 763.0a 828.8a
For each chemical property of soil mixtures, lines of table with the same letter do not
differ significantly according to the Tukey‟s test (P=0.05). C, control (50g soil plus
9.84g manure); R1, R2 or R3, control and residue robusta 0.3, 0.6 or 0.9g,
respectively.

CONCLUSIONS
Three different rates of green coffee residue, obtained after the removal of the
extract with hot water to determine the antioxidant activity, were applied into a soil
amended with manure. These results, confirm the role of green coffee residue as it
could be applied to the soil as an acceleration agent for soil organic matter
biodegradation, without any extremely negative effect on the available forms of
nutritional mineral elements, while that recycling constitutes a useful practice for
environment protection.

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