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Mitigation of Salt Stress by Organic Matter and GA3 on Growth and Peroxidase
Activity in Pepper (Capsicum annum L.)

Article · August 2017

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 ADVANCES in NATURAL and APPLIED SCIENCES
ISSN: 1995-0772 Published BYAENSI Publication
EISSN: 1998-1090 http://www.aensiweb.com/ANAS
2017 August 11(10):pages 1-11 Open Access Journal

Mitigation of Salt Stress by Organic Matter

and GA3 on Growth and Peroxidase Activity
in Pepper (Capsicum annum L.)
D. K.A. Altaey

Assistant Prof. in Plant Physiology Horticulture Department, Agriculture collage /Al-Qasim Green University -Babylon-Iraq

Received 22 May 2017; Accepted 25 July 2017; Available online 10 August 2017

Address For Correspondence:

Duraid K.A.Altaey, Assistant Prof. in Plant Physiology Horticulture Department, Agriculture collage /Al-Qasim Green University
Babylon-Iraq

Copyright © 2017 by authors and American-Eurasian Network for Scientific Information (AENSI Publication)
This work is licensed under the Creative Commons Attribution International License (CC BY).
http://creativecommons.org/licenses/by/4.0/

ABSTRACT
Background: Capsicum is one of the most widely consumed vegetables and is also used as a spice for its pungency. Many species
of Capsicum are being cultivated worldwide. Capsicum are considered as a commercial crop for their economical value. However,
the yield of the crop suffers severely due to salt stress, Soil salinity reduces water availability of plant roots via negative (low)
osmosis potential, as well as decrease of germination dynamics of plant seeds by ionic toxicity of Na and Cl, Significant differences
in fruit-set, yield, photo synthetic rates, stomatal conductance, total chlorophyll content, proline, In general, salinity affects almost
every aspect of the physiology and biochemistry of plants. Objectives: The aim of this study was to determine the salt tolerance of
pepper (Capsicum annuum L) under salinity stress by saline irrigation water, Poultry and gibberellins applications were used to
alleviated the negative effects on growth parameters and yield of Pepper under salinity stress. Results: The water salinity levels
led to a significant elevation in the values of electrical conductivity of the soil with the peroxidase activity, and Sodium and proline
contents in leaves, while resulting in decrease in growth parameters and leave contents of ( NPK),The poultry and gibberellins
applications increased the growth parameters ( Dry weight of shoot and root &fruit weight) and (NPK) contents in leaves with
slight dropping of peroxidase activity in leaves while a clear dropping of sodium and proline contents in leave. Conclusions: That
possible to mitigation the negative effect of salt stress by some application like exogenous hormones and Decomposed organic
matter to solve the disruption of endohormons and lack of available nutrients under salt stress, and elevation of osmotic stress in
soil solution in roots area. The GA & poultry application improved the growth and it has increased the Pepper tolerance to the
abiotic stress which was exerted by saline irrigation water.

KEYWORDS: salinity, salt stress, pepper, Gibberellins, organic matter, poultry manure, nutrient availability, osmotic pressure

INTRODUCTION

Growth and productivity of the plants are affected due to many abiotic stresses like salinity, heat, cold and
drought etc.[1] which are leading towards hundreds of billions of crop losses each year [2]. Soil salinity is the
most devastating among them [3], which not only limits plant growth and metabolism but also poses a foremost
intimidation to sustainable agricultural production throughout the world particularly in arid and semi-arid areas
[4]. More than 400 million hectares of the total geographical area of the world are affected by high
concentration of the soluble salts [1] Salt stress severely inhibits plant growth for two reasons: first by an
osmotic or water- deficit effect of salinity and second by a salt-specific or ion-excess effect of NaCl. Moreover,
plants subject to salinity stress conditions produce cytotoxic activated oxygen that can seriously disrupt normal
metabolism, through oxidative damage of lipids, proteins, and nucleic acids [5], Salinization can also lead to
excess intracellular production of reactive oxygen species (ROS) such as the superoxide radical (O2 ·–), the
hydroxyl radical (OH·), hydrogen peroxide (H2O2) and singlet oxygen (1O2) [6].The tolerance of salinity is
ToCite ThisArticle:Duraid K.A.Altaey.,Effects of Water Salinity ,Poultry and Gibberellins Applications on Growth of Selected
Nutrients and Proline Contents, with Peroxidase Activity in Pepper (Capsicum annum L.).Advances in Natural and Applied
Sciences. 11(10);Pages: 1-11
2 Duraid K.A.Altaey, 2017/Advances in Natural and Applied Sciences. 11(10) August 2017, Pages: 1-11

specific for each species or cultivar. Vegetables have a high sensitivity to the effects of NaCl [7], which hinders
growth because of its toxic and osmotic effects, respectively, causing accumulation of ions in the protoplasm
and physiological drought; [8]. To defend against such oxidants, plants have evolved specific protective
mechanisms, involving antioxidant molecules and enzymes that protect against the potentially-cytotoxic species
of activated oxygen. Adaptation to salt stress requires alterations in gene.
Pepper (Capsicum annuumL.) is the second most widely consumed vegetable in the world and an excellent
source of many essential nutrients for humans, especially vitamin C, phenolic compounds, flavonoids,
tocopherols (vita-min E), carotenoids (pro vitamin A), capsaicinoids, and calcium. Additionally, some pepper
cultivars contain significant quantities of capsaicinoids, a group of pungent phenolic derived compounds with
strong physiological and pharmacological properties. Thus, the growing global demand of pepper fruits implies
several strategies to increase crop production and fruit quality or promote the investigation to improve the plant
resistance to environmental stresses [9], Pepper is a moderately sensitive to salt stress [10], and it is grown
under protected glasshouse conditions in temperate regions and in the open field under warm Mediterranean
climates, it is frequently exposed to saline conditions brought about by saline irrigation water containing
amounts of salts including sodium chloride [11]
Salinization promotes an imbalance in the absorption of essential nutrients, causing metabolic disorders,
which inhibit growth [12], there are an extensive number of plant nutrition studies from all over the world, but
the studies were mostly conducted to determine best management practices under non-saline conditions. Some
studies have been conducted to determine if certain nutrients have alleviative effects on salinity tolerance [13].
Some studies indicated a positive effect of fertility on salt tolerance while some reported that there was no
alleviative effect on salt tolerance, some Studies showed that application of fertilizers in saline soils might result
in increased, decreased or unchanged plant salt tolerance. In other words, plant response to fertilizers depends
on severity of salt stress in the root zone [14],However, in another similar study to [15], found a positive yield
response for pepper at all three salinity levels by increasing nutrient N from 2 to 15 mM in a solution culture.
However the effect of N on relative yield was not clear. The first salinity level above the control (25 mM NaCl)
had a lower relative yield at lower N and with subsequent increases in salinity it had a higher relative yield.
phytohormones are considered the most important endogenous substances for modulating physiological and
molecular responses, a critical requirement for plant survival as sessile organisms, Phytohormones act either at
their site of synthesis or elsewhere in plants following their transport [16].
The gibberellins (GAs) are a large group of tetracyclic diterpenoid carboxylic acids, The GAs show
positive effects on seed germination, leaf expansion, stem elongation, flower and trichome initiation, and flower
and fruit development, They are essential for plants throughout their life cycle for growth-stimulatory functions.
They also promote developmental phase transitions. Interestingly, there is increasing evidence for their vital
roles in abiotic stress response and adaptation[17]and [16].Recently, experiments have been performed to
investigate the role of GAs in osmotic stress response in Arabidopsis thaliana seedlings [18].[19] reported that
GA3 treatment in tomato reduced stomatal resistance and enhanced plant water use at low salinity. Likewise,
GA3-priming increases grain yield due to the GA3-priming-induced modulation of ion uptake and partitioning
(within the shoots and roots) as well as hormone homeostasis under saline conditions.
GAs are known to interact with all other phytohormons in numerous developmental and stimulus-response
processes, The interactions between GA and ET include both negative and positive mutual regulation depending
on the tissue and signaling case [20].

MARTIAL ANDMETHODS

This experiment was conducted under glass house of horticulture department, collage of in AL- Qasim green
university at Novemb1st 2015, the Sweet pepper (Capsicum annuum L.) of RIDA cultivar from Netherland
was used. The seedlings were planted in plastic pots containing 10 kg of soil (six pots for each treatment).Each
one supplied with 0.5 gm of NPK and granular fungicide. Seedlings were irrigated with river water (1.2 dS.m-1
/cm) for ten days twice a day before salinity treatment, followed by irrigation (half of seedlings) with saline
water (6 dS.m-1 /cm) every day until seedlings were reaching 80 days old. Plants were sprayed twice with of
GA (0, 250 mg /L) the first spray was two weeks after germination, the second spray was 4 weeks after the first
spray. Experiment was conducted according to split-split plot design with threefactors, The main factor is the
water quality (1.2 dS.m-1 represented river water (W1) & 6 dS.m-1 represented saline water (W2),the second
factor (sub- plot) is the poultry fertilization levels with 10% (O1) &30% (O2),The third factor (sub-sub-plot) is
gibberellin levels with (0, 250 mg/liter)The Gibberellin 0% (G1) & the 250 mg/liter (G2.) ,the data were
analyzed statistically with Genstat discovery software. Means were statistically compared by L.S.D testat
p<5%[21], the nitrogen determination according to [22]while the determination of phosphorus in leaves was
measured according to [23].
3 Duraid K.A.Altaey, 2017/Advances in Natural and Applied Sciences. 11(10) August 2017, Pages: 1-11

Determination of Peroxidase Activity:

This was determined by measuring the increase in absorbance at 510 nm resulting from the decomposition
of hydrogen peroxide [24],The Lambda 25 UV/Vis spectrometer (Perkin Elmer) was adjusted to 510 nm. The
blank was a mixture of 1.4 ml of phosphate buffer and 1.4 ml of H2O2 in the cuvette. The assay mixture
contained 1.4 ml of phosphate buffer, 1.4 ml of H2O2 and 0.2 ml of the extract. The increase in absorbance at
510nm was recorded for 4 minutes. Then, ΔA240/min was calculated from the initial (45 second) linear portion
of the curve.

Determination of proline:
Proline colorimetric determination preceded according to [25]&[26] based on proline's reaction with
ninhydrin ratio of 1:1:1 solution of proline, ninhydrin acid and glacial acetic acid was incubated at 100ºC for 1
hour. Thereaction was arrested in an iced bath and the chromophore was extracted with 1 ml toluene and its
absorbance at 520 nm was determined spectrophotometerically .0.1 gm of shoot and root tissues was suspended
with 1 ml of 3% sulfosalicylic acid and after centrifugation (10 min at 12,000 rpm) was mixed in a 1:1:1 ratio
with ninhydrin acid and glacial acetic acid. The reaction and determination of proline were carried out similarly
tothat described above The concentration of proline in tissues were determined dependingon standard curve of
pure proline.

Results:
1- Dry weight of shoot and root, fruit weight & chlorophyll content in leaves:
The figures (1,2,3 & 4) show a significant effect of poultry manure at 25% concentration on dry weight of
shoot and root, fruit weight and chlorophyll content in leaves with boost rat was (130%,93%, 99% & 13%),
sequentially according to 0% of poultry manure concentration, In a similar manner to gibberellin application of
250 mg /liter with boost rate was (31%,42%, 84% & 14%), sequentially according to 0 mg/liter of gibberellin
concentration, but there is significant drop to dry weight of shoot and root &fruit weight with raising of water
salinity,the dropping rat was (45%, 34% , 58% & 7%) g, sequentially, the interaction treatment among (poultry
25% +gibberellin 250 mg + irrigation water 1.2 ds/m) achieved the highest means in dry weight of shoot and
root, fruit weight & chlorophyll content in leaves while the lowest means at treatment (poultry 0 %, gibberellin
0%,irrigation water 6ds/m),the application of poultry manure and gibberellin treatment alleviated the negative
affect of saline water in dry weight of shoot and root, fruit weight &chlorophyll content with (3.76, 2.11, 21.9&
49.9), sequentially according to treatment which irrigated by saline water without poultry and gibberellin
application which recorded (1.48, 0.65 , 9.8 & 44.9) sequentially.

Fig. 1: shows the effect of water quality, poultry manure& gibberellin with interaction between them on the dry
weight of leaves. {Water quality (W) Poultry manure (O) Gibberellin (g)}
4 Duraid K.A.Altaey, 2017/Advances in Natural and Applied Sciences. 11(10) August 2017, Pages: 1-11

Fig. 2: shows the effect of water quality, poultry manure& gibberellin with interaction between them on the dry
weight of root. {Water quality (W) Poultry manure (O) Gibberellin (g)}

Fig. 3: shows the effect of water quality, poultry manure& gibberellin with interaction between them on the
fruit weight. {Water quality (W) Poultry manure (O) Gibberellin (g)}
5 Duraid K.A.Altaey, 2017/Advances in Natural and Applied Sciences. 11(10) August 2017, Pages: 1-11

Fig. 4:shows the effect of water quality, poultry manure& gibberellin with interaction between them on the
chlorophyll content in spad. {Water quality (W) Poultry manure (O) Gibberellin (g)}

2- The Nitrogen, Phosphor, Potassium and Sodium in the leaves:

The figures (10 & 11) show a significant difference from treatments with proline contents of root and
peroxidase activity in leaves, the poultry manure 25% achieved significant values where boost rate was (9% &
13%) sequentially compared with treatment which none poultry manure fertilize, similarly the Gibberellin
application by 250 mg /liter produced a significant increments with proline contents of root and peroxidase
activity in leaves compared with treatment none Gibberellin treats, and peroxidase activities figures (10 & 11)
shows reductive effect of saline water with dropping rate if (78% & 49%) sequentially compared with river
water, these results agreed with [27;28] which they find that there elevation of proline contents in leaves with
raising of water salinity.
The tertiary interaction shows a significant affect amongst treatments of proline contents in leaves, the best
result was (W2G2O1) & (W2G2O1) with (7.81, 7.58) mmole .g-1, sequentially and the lowest result at the
treatment (W1G2O2) with (3.88) mmole .g-1.and the best results of peroxidase activity was (W2G2O1) &
(W2G2O2) with (102.67, 91.33)mg of protein -1 sequentially, and the lowest results at the treatment (W1G2O2)
with (54.33) mg of protein -1 figures (10,11). The application of poultry manure and gibberellins reduced the
negative affect of salinity by saline water, the treatments (W2G1O1) recorded (7.58) mmole .g -1 of proline and
(102.67) mg of protein -1 of peroxidase activity compared with (W1G2O2) treatment which recorded (3.88)
mmole .g-1 and (54.33) mg of protein -1 sequentially.

Fig. 5: shows the effect of water quality, poultry manure& gibberellin with interaction between them on the N
content in leaves. {Water quality (W) Poultry manure (O) Gibberellin (g)}
6 Duraid K.A.Altaey, 2017/Advances in Natural and Applied Sciences. 11(10) August 2017, Pages: 1-11

Fig. 6:shows the effect of water quality, poultry manure& gibberellin with interaction between them on the
Phosphorus content in leaves. {Water quality (W) Poultry manure (O) Gibberellin (g)}
%

Fig. 7: shows the effect of water quality, poultry manure& gibberellin with interaction between them on the K
content in leaves. {Water quality (W) Poultry manure (O) Gibberellin (g)}
7 Duraid K.A.Altaey, 2017/Advances in Natural and Applied Sciences. 11(10) August 2017, Pages: 1-11

Fig. 8: shows the effect of water quality, poultry manure& gibberellin with interaction between them on the Na
content in leaves. {Water quality (W) Poultry manure (O) Gibberellin (g)}

Fig. 9 : shows the effect of water quality, poultry manure& gibberellin with interaction between them on the
K/Na content in leaves. {Water quality (W) Poultry manure (O) Gibberellin (g)}

3- Proline content in root & Peroxidase activity in leaves:

The figures (11 & 12) show a significant difference from treatments with proline contents of root and
peroxidase activity in leaves, the poultry manure 25% achieved significant values where boost rate was (9% &
13%) sequentially compared with treatment which none poultry manure fertilize, similarly the Gibberellin
application by 250 mg /liter produced a significant increments with proline contents of root and peroxidase
activity in leaves compared with treatment none Gibberellin treats, and peroxidase activities figures (11 & 12)
shows reductive effect of saline water with dropping rate if (78% & 49%) sequentially compared with river
water.
The tertiary interaction shows a significant affect amongst treatments of proline contents in leaves, the best
result was (W2G2O1) & (W2G2O1) with (7.81, 7.58) mmole .g -1, sequentially and the lowest result at the
treatment (W1G2O2) with (3.88) mmole .g-1.and the best results of peroxidase activity was (W2G2O1) &
 8 Duraid K.A.Altaey, 2017/Advances in Natural and Applied Sciences. 11(10) August 2017, Pages: 1-11

(W2G2O2) with (102.67, 91.33)mg of protein -1 sequentially, and the lowest results at the treatment (W1G2O2)
with (54.33) mg of protein -1 figures (11,12).
The application of poultry manure and gibberellins reduced the negative affect of salinity by saline water,
the treatments (W2G1O1) recorded (7.58) mmole .g -1 of proline and (102.67) mg of protein -1 of peroxidase
activity compared with (W1G2O2) treatment which recorded (3.88) mmole .g -1 and (54.33) mg of protein -1
sequentially.

Fig. 10: shows the effect of water quality, poultry manure& gibberellin with interaction between them on
Proline content mmole .g-1 in root. {Water quality (W) Poultry manure (O) Gibberellin (g)}

Fig. 11: shows the effect of water quality, poultry manure& gibberellin with interaction between them onPeroxidase
activity in mg of protein -1of leaves. . {Water quality (W) Poultry manure (O) Gibberellin (g)}

Discussion:
the figure (1,2,3,4,5,6,7,8 ) show a significant reduction in dry weight of shoot and root, fruit weight,
chlorophyll contents of leaves,Nitrogen,phosphorus, Potassium & K/Na,when used saline water compared with
river water,similarly [29] showed that salt stress was negatively affect wet weight, leaves number; leaves
surface area and shoot length, The inhibitory effects of salinity on growth of pepper plant the effects of high soil
salt availability and are probably due to decreased water absorption and disturbed metabolic processes leading
to decreased meristematic activity or cell enlargement [30]. [31] Reported that there are two ways which
salinity could retard growth, by damaging growth cells so that they cannot perform their functions or by limiting
 9 Duraid K.A.Altaey, 2017/Advances in Natural and Applied Sciences. 11(10) August 2017, Pages: 1-11

their supply of essential metabolites. Salinity stress is known to retard plant growth through its influence on
several vital factors of plant metabolism, including osmotic adjustment [32],nutrient uptake, protein and nucleic
acid synthesis, photosynthesis [33], organic solute accumulation, enzyme activity, hormonal balance and
reduced water availability at the cell level all of which result in reduced plant growth and ultimately reduced
yield. Furthermore, increased salt content in the irrigation water may cause direct and indirect effects on leaf
water relations and stomata closure which influence CO2 exchange and photosynthetic rate. Increased salt
content in irrigation water may be directly toxic to plants, which in turn, lowered carbohydrate accumulation in
the plants [34; 35]
The, proline contents in root and peroxidase activity were increased with saline water figure (10, 11)
respectively, these are one of the role which plant followed to scavenge the reactive oxygen species, the
effects of salt stress on plant growth to an increase in reactive oxygen species which play an important role in
damaging all classes of biologically important macromolecules including DNA and the generation of H2O2 and
lipid hydro-peroxides which cause membrane changes, To mitigate and repair damage initiated by reactive
oxygen, plants have developed a complex antioxidant system. The primary components of this system include
some enzymes such as peroxidase (POX), catalase (CAT) super oxidase dismutase (SOD) and proline [36; 37].
The poultry and gibberellins applications alleviated from the negative affect of saline water figures
(1,2,3,4,5,6,7,8). Organic fertilizer apart from releasing nutrient elements to the soil has also been shown to
improve other soil chemical and physical properties which enhance crop growth and development [38],In
addition, poultry manure has also been reported to increase soil pH, hence the acidic soil of the experimental
site which could have caused the unavailability of nutrient element to the crops was checked by the limiting
potential of organic manure [39], Moreover, poultry manure contains essential nutrient elements associated with
high photosynthetic activities and thus promoted roots and vegetative growths [40],gibberellin play vital role in
regulating developmental processes within plant bodies [41]. Gibberellin helps in cell growth of stem, leaves
and other aerial parts by causing cell elongation, and increase in intermodal length. A higher concentration of
gibberellins increases plant growth [42].
[43].reported that treatment of GA causes stem elongation, expansion and proliferation and cell wall
thickening in bast fiber of linseed, GA3 counteracts with salinity by improving membrane permeability and
nutrient levels in leaves which ultimately leads to better growth and also GA3 induced physiochemical changes
responsible for induction of salt tolerance [44].
Knowledge:
That possible to mitigate the negative effects of salt stress by some application like exogenous hormones
and Decomposed organic matter to solve the disruption of endohormons and lack of available nutrients under
salt stress, and Osmotic stress elevation in soil solution in roots area.
We recommended that more researches about salt stress in arid and semi-arid zones to be conducted and the
use of other applications from sources of organic matters with studying the phytohormones (Auxins
gibberellins, cytokinis, ethylene,ABA, etc.. in addition to studying the interactions between them on growth and
yield to other plants for discovering and increasing the plant tolerance.

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