Scientia Horticulturae 225 (2017) 581588

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Scientia Horticulturae
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Original Research Article

Gibberellic acid coating: A novel approach to expand the shelf-life in green MARK
chilli (Capsicum annuum L.)

Jitendriya Panigrahia, , Bhumi Gheewalaa, Mansi Patela, Niyati Patela, Saikat Gantaitb,c,
a
Department of Biotechnology, Shri A. N. Patel P.G. Institute of Science and Research, Anand, Gujarat 388001, India
b
All India Coordinated Research Project on Groundnut, Directorate of Research, Bidhan Chandra Krishi Viswavidyalaya, Kalyani, Nadia, West Bengal 741235, India
c
Department of Genetics and Plant Breeding, Faculty of Agriculture, Bidhan Chandra Krishi Viswavidyalaya, Mohanpur, Nadia, West Bengal 741252, India

A R T I C L E I N F O A B S T R A C T

Keywords: In the present study, for the rst time, we report the eectiveness of gibberellic acid (GA3) coating on the
Capsicum annum expansion of postharvest shelf-life in green chilli (Capsicum annum L.). The fruits were coated with GA3 dipping
Coating in 1, 2 and 3 ppm for 30 s and then stored at 4 1 C. Fruits without any GA3 coating were considered as
GA3 control. All the treated and control fruits were stored as such for 45 days and throughout the storage period (with
Postharvest
every nine days intervals) the multiple enzyme and antioxidants (associated with storage) such as titratable
Shelf-life
acidity, ascorbic acid content, ferrous ion chelating activity, reducing power, 2,2-diphenyl-1-picrylhydrazyl
(DPPH) scavenging activity, hydroxyl radical scavenging activity, total phenolic content and enzymatic study of
polyphenol oxidase and pectate lyase activity were assessed. The fruits coated with all three levels of GA3
showed a signicant delay in change of skin colour, decline in titratable acidity, total phenolic and ascorbic acid
content, increase in enzyme and antioxidant activities during the cold-storage in comparison to the control fruits.
GA3, used as the coating in green chillis, eventually showed benecial roles in decelerating the mechanism of
ripening. The results further indicated that the 2 ppm of GA3 concentration served superiorly than the 1 ppm or
3 ppm throughout storage and provided expanded shelf-life of green chilli.

1. Introduction dihydrocapsaicin and capsaicin (Hornero-Mndez et al., 2002). Being a

Preservation of natural products for an extended period without any
deterioration is a universal requirement. Every fruit or vegetable have
their own life span, yet as per our need, we want to make them avail-
able for more than the expected period. Multiple preservatives are
being used to serve this purpose. Nevertheless, some of them were not
preferable to us due to their residual side-eects, instead, the usage of
coatings that can delay the degradation process and extend the shelf-life
are preferred.
Green chilli (Capsicum annuum L.) is a member of Solanaceae.
Though it is originated from South and Central America it is widely
used as an ingredient in cooking for its spicy nature throughout the
world. It consists of various chemicals like steam volatile oil, fatty acid,
vitamin A, vitamin C, vitamin E, proteins, bres and mineral elements
like potassium and folic acid (Sitthiwong et al., 2005). Two distinctive
compounds, capsanthin and capsaicin are responsible for red colour and
pungency of chilli. The major percent of total capsaicinoids are
non-climacteric fruit, the postharvest deterioration of green chilli is
very rapid that results into huge losses. Due to high moisture content
(6085%) during harvest (Charmongkolpradit et al., 2010) the green
chillies are prone to fungal attack. Degradation of quality, shrivelling
with the loss of weight, change in skin colour and chilling injury when
stored at below 7 C are the common issues associated with postharvest
storing and handling (Nyanjage et al., 2005). These complications
eventually turn green chilli as a highly perishable vegetable (Xing et al.,
2011). Yet, there are very limited attempts have been made on the
shelf-life extension of green chilli in comparison to bell pepper
(Chitravathi et al., 2015).
Researches suggest that organic or inorganic coatings applied in ne
layers on the fruit skin as a supplement or substitute to the natural waxy
overlays act as a barrier to moisture, oxygen, and passage of solutes in
the fruit. By dipping, spraying, or brushing the coatings are applied
directly on the food surface to make an improved state for storage
(Quirs-Sauceda et al., 2014). Coatings have a good bond to rough

Corresponding author at: All India Coordinated Research Project on Groundnut, Directorate of Research, Bidhan Chandra Krishi Viswavidyalaya, Kalyani, Nadia, West Bengal 741235,
India.

Corresponding author.
E-mail addresses: jitenp2001@gmail.com (J. Panigrahi), bhumigheewala93@gmail.com (B. Gheewala), maansipatel1512@gmail.com (M. Patel), niyaa_001@yahoo.com (N. Patel),
saikatgantait@yahoo.com (S. Gantait).

http://dx.doi.org/10.1016/j.scienta.2017.07.059
Received 16 April 2017; Received in revised form 29 July 2017; Accepted 31 July 2017
0304-4238/ 2017 Elsevier B.V. All rights reserved.
J. Panigrahi et al.
surfaces (Hershko et al., 1996). An ideal coating is distinct since it
extends the storage life of fresh fruits and vegetables without causing
anaerobiosis and reduces decay without aecting their quality (Jyand
et al., 1994; Park et al., 1994a). The role of coatings on product surface
relies on temperature, thickness, acidity, and the variety and nature of
coating including condition of fruit and vegetable (Park et al., 1994b).
Such coatings possess immense capacity to transmit active ingredients,
e.g. anti-browning agents, colours, essence, proteins, vitamins and mi-
nerals, and antimicrobial component that can prolong the shelf-life of
product and minimise the menace of pathogenic contamination on food
surface (Quirs-Sauceda et al., 2014). The ecacy of coatings depends
on the molecular structure, molecular size and chemical composition.
The gibberellic acid (GA3) proved to be a successful agent that
signicantly delayed the ripening of several fruits and extended shelf-
life simultaneously. It was observed that both at 25 C and 15 C GA3,
reportedly checks the breakdown of ascorbic acid and chlorophyll in
the skin, and lessens -amylase and peroxidase activities throughout
storage (Khader, 1992). Furthermore, supplementation of GA3 at low
temperature eectively reduces soluble protein and sugar, decreases the
action of polyphenol oxidase and peroxidase plus accumulation of free
amino acids during storage that eventually extends shelf-life of fruits
(Tian et al., 2014). Even though, GA3 has been used as an agent that
positively inuenced the length of shelf-life without compromising the
quality in few fruits and vegetables such as mango, banana, peach,
iceberg lettuce and plums (Khader, 1992; Kumar, 1998; Osman and
Abu-Goukh, 2008; Dagar et al., 2012; Huang et al., 2014; Tian et al.,
2014; Erogul and Sen, 2016) but there is scanty of report on its use as a
coating in green chilli. Hence, in the present study for the rst time, we
report the extension in shelf-life of green chilli with the application of
GA3 as coating agent and assess its interaction with some storage-linked
enzymes and antioxidants.
2. Materials and methods
2.1. Fruit collection, preparation and application of coating
Green chilli fruits, used as the experimental material, were collected
from a local market near Anand, Gujarat, India. The fruits were washed
with tap water to remove the adhered dirt on their skins. For further
sterilization of the surface, the fruits were then dipped in 70% (v/v)
ethanol for 30 s and were thoroughly rinsed with sterile-distilled water.
Prior to their storage in common refrigerator (at 4 1 C), the fruits
were treated with 1, 2 and 3 ppm of GA3 solution for 30 s and after a
complete drying, the fruits were wrapped in aluminium foil. The fruits
without any GA3 treatment, considered as control, were also wrapped
in aluminium foil and stored in refrigerator. The control and GA3-
treated fruits were then observed for 45 days and throughout the sto-
rage period (with each nine days intervals) the multiple enzymes and
antioxidants were assessed (described below) along with the estimation
of change in skin colour.
2.2. Assessment of skin colour
The change in skin colour following dierential storage period was
assessed with the help of RHS standard colour chart (Fifth Edition.
The Royal Horticultural Society London, 2007).
2.3. Assessment of titrable acidity
To prepare the homogenate, the chilli fruits were peeled and
chopped randomly. Five grams of such fruit pulp was taken and mixed
well with 50 ml distilled water and then centrifuged at 5000 rpm for
10 min. Titratable acidity was determined by titration of 20 ml of chilli
homogenate (supernatant) with 0.1 N NaOH up to pH 8.1. Three re-
plicates were used every two days for each treatment.
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Scientia Horticulturae 225 (2017) 581588
2.4. Assessment of ascorbic acid content
Ascorbic acid level was determined following the protocol of
Ranganna (1986). The chilli fruits were peeled and chopped. Such 10 g
of fruit pulp was crushed and diluted with 100 ml distilled water. Ten
millilitre of this homogenate of chilli fruit was dissolved with 10 ml of
20% meta phosphoric acid in a mortar and pestle. The solution was
then collected in a 100 ml volumetric ask and made up to the volume
of 100 ml with distilled water. Titration was made with the standard
2,6-dichlorophenol indophenol dyes. The ascorbic acid level of in-
dividual samples was calculated by the formula: Ascorbic acid (mg/
100 ml homogenate i.e. mg/10 g fruit pulp) = (Titre dye factor -
volume made up 100)/(volume taken for titration sample
weight). The level of ascorbic acid was designated as mg/10 g fruit
pulp.
2.5. Total soluble sugar estimation
The estimation of total soluble sugar was measured by taking
100 mg of plant samples followed by Yemm and Willis (1954).
2.6. Assessment of ferrous ion chelating activity
The chelating eect was measured following the method of Shimada
et al. (1992). Ethylene diamine tetraacetic acid (EDTA) was taken as a
control. The ferrous ion chelating activity was measured via the amount
of decline in absorbance ratio in the presence of polysaccharide and
expressed as millimolal (mm) of chelted ferrous ion.
2.7. Assessment of reducing power
Five grams of the chilli fruits were ground with 50 ml of distilled
water and centrifuged at 5000 rpm for 10 min and the supernatant was
collected. Serial dilution of these extracts (200, 100, 50, 25 and
12.5 g/ml) was carried in 0.2 M phosphate buer pH 6.6 comprising
1% ferrocyanate. After incubation at 50 C for 20 min, 10% tri-
chloroacetic acid (TCA, 2.5 ml) was mixed with a part (5 ml) of this
mixture and centrifuged at 3000g for 10 min. The supernatant was
isolated and diluted with distilled water (2.5 ml) having 1% ferric
chloride (0.5 ml); and then estimated at 700 nm. The intensity in ab-
sorbance could be the measurement of antioxidant activity of the ex-
tract (Yen and Duh, 1994)
2.8. Assessment of total phenolic content
The total phenolic content was assessed following the method pro-
posed by Singleton et al. (1998). For the calibration curve, aliquots
(1 ml) of 0.024, 0.075, 0.105 and 0.3 mg/ml of gallic acid methanol
solutions were blended with Folin-Ciocalteu reagent (5 ml, diluted ten
fold) and sodium carbonate (75 g/l, 4 ml). After 30 min, the absorption
was recorded at 765 nm at 20 C and the calibration curve was outlined.
For extraction of polyphenol a mixture of 10 g ne homogenate
(chrushed fruit pulp) of chilli fruit with 100 ml acidic methanol was
prepared and kept at 4 C that was ltered through ordinary lter paper
later on. One militre of such chilli fruit extract was mixed with the
similar reagents as stated above, and after 1 h the absorption was re-
corded for the assessment of total phenolics. The total level of phenolic
compounds in fruit extracts in gallic acid equivalents (GAE) was de-
termined following the formula of: C = c. V/m, where C is the total
content of phenolic compounds, mg/g extract, in GAE; c is the con-
centration of gallic acid ascertained from the calibration curve, mg/ml;
V is the volume of extract in ml; m is the weight of pure methanol
extract in g (Miliauskas et al., 2004). The results were expressed as mg
GAE/g extract.
J. Panigrahi et al.
2.9. Assessment of hydroxyl radical scavenging activity
Hydroxyl radicals were produced by a Fenton reaction (Fe3+- as-
corbate-EDTA-H2O2 system), and the scavenging activity towards the
hydroxyl radicals was assessed following the deoxyribose method (Kaur
and Halliwell, 1994).
2.10. Assessment of 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical
scavenging activity
The peeled chilli fruits were dried and lyophilized. The lyophilized
chilli fruits were powdered and 50 gm of this powder was suspended in
500 ml milipore water and extracted with 85% ethyl acetate at room
temperature for 12 h (3 times with 500 ml). The ethyl acetate fraction
was separated and the remaining aqueous part was treated as sub-
fractions. The antioxidant activity of sample ethyl acetate fractions,
subfractions and the antioxidant standard were evaluated based on the
radical scavenging eect of the stable DPPH free radical (Shimada
et al., 1992).
2.11 Assessment of polyphenolic oxidase activity
The enzymatic activity was determined by measuring the increase in
optical density. A 0.01 M catechol solution (2.5 ml) and 22 ml of a
0.05 M phosphate buer (pH 6.0) were mixed and kept at 30 0.5 C
583
Scientia Horticulturae 225 (2017) 581588
Fig. 1. Eect of GA3 coating and storage period on skin colour of
green chilli (Capsicum annuum L.).
for 10 min in a water bath. Five grams chilli fruit pulp was homo-
genized with 50 ml of 0.05 M potassium phosphate buer (pH 6.8) and
centrifuged at 8000 rpm for 15 min. The supernatant was treated as
enzyme extract. The enzyme extract (0.5 ml) was added to the above
mixture with further mixing achieved by swirling. The optical density
was then measured at 430 nm. A reference cuvette containing the sol-
vent was placed in the sample and solvent compartments and the col-
orimeter was zeroed until a zero absorbance was displayed. The sample
cuvette was then used for measurements on the samples. At intervals of
two min absorbency readings were made from the reacting mixture.
Duplicate determinations were carried out and the average result re-
corded. PPO activity was obtained by determining the initial slope of
the curve obtained by plotting optical density at 430 nm against time in
min. The results were expressed as the increase in OD/min/ml of
enzyme solution (Luh and Phithakpol, 1972).
2.12 Assessment of pectate lyase activity
The pectate lyase enzyme was extracted by homogenising 5 gm of
chilli fruit pulp in 75 ml cold 8.8% NaCl and centrifuged at 20,000 G for
10 min. The supernatant was collected and the pH adjusted at 7.5 with
NaOH. Then 2 ml of pectin was mixed with 0.15 ml bromothymol blue
and 0.83 ml distilled water. The mixture was incubated at 25 C with
circulating water bath. The enzymatic reaction was started by adding
100 l enzyme solution and the absorbance was observed at 620 nm
J. Panigrahi et al.
after 20, 40, 60 and 80 s intervals. One unit (U) of pectate lyase activity
is the amount of enzyme causing a change in absorbance of 0.01 under
the conditions of the assay (Moran et al., 1968).
2.13 Statistical analysis
The experiments were executed following Completely Randomized
Design (CRD). Each experiment was replicated three times with 30
samples per replication. Recorded data were statistically assessed using
two-way analysis of variance (ANOVA). To assess the least signicant
dierence among the treatments, the data (means SD) were then
calculated at P < 0.05 using SPSS (Version 20, SPSS Inc. Chicago,
USA) software.
3. Results and discussion
3.1. Skin colour
The fruits stored followed by GA3 coating showed dierential re-
sponse on their skin colour when they were photographed at certain
interval under storage. The control fruits turned into red colour (code
44B) with necrotic tip signicantly earlier (following 18 days of sto-
rage) than the fruits with GA3 coating and eventually rotten up with
red-orange colour (code 34D) following 27 days of storage. The fruits
with 2 ppm GA3 coating were the last ones to be turned into yellow-
green (code 144C) or green (141C) with red (code between 40A and
40B) tinge, whereas 1 ppm and 3 ppm GA3 coating turned the skins
from green (code between 141A and 141B) to red (code 43A) or ne-
crotic and yellow-green (code between 143 B and 143C) with red (code
44A) tinge much earlier than that of the 2 ppm (Fig. 1). Throughout
postharvest storage green chilli experience intense transformations and
conversion of existing pigments. Existence of chlorophyll is the key
reason of green colour of the fruit whereas degradation of chloroplast
pigments and exclusive synthesis of carotenoid chromoplast pigments
and their esterication by fatty acids occurs during maturation and
senescence (Hortensteiner, 2006). However, the slow transformation of
skin colour and long-term retention of green colour in the fruits with
2 ppm GA3 coating might be due to reduced respiration rate caused by
optimized GA3 coating. According to Chitravathi et al. (2015) slow
respiration rate ensued in limited degradation of chlorophyll and re-
stricted synthesis rate of carotenoids, which occur during senescence. In
the present study, transformation of skin colour from green to yellow to
red-orange during ripening might be attributed to the degradation of
chlorophyll pigment and parallel biosynthesis of carotenoids (Deepa
et al., 2007). Comparable outcomes were also detected in green chilli
by Chitravathi et al. (2015).
3.2. Titrable acidity
Organic acids like citric acid are the prime substrates for respiration;
a decrease in acidity and a rise in pH are liable in highly respiring fruits.
Such coatings minimize respiration rates and hinder the activities of
organic acids (Yaman and Bayindirli, 2001). During the storage period
in our study, the outcomes revealed that titratable acidity values in all
cases were slowly and considerably (signicant at P < 0.05) lowered
with increasing storage period. At the end of the 18-days storage
period, the control samples showed its least titratable acidity
(0.127 0.006% citric acid) and following which the control samples
were deteriorated completely. At the same storage period (18 days), the
highest value of titratable acidity was shown by chillies treated with
2 ppm GA3 (0.307 0.006% citric acid) followed by the 3 ppm ones
(0.223 0.006% citric acid) (Table 1). Nevertheless, the shelf-life of
chilli was also observed to be extended up to 45 days for the fruits
treated with 2 ppm GA3. The two-way ANOVA for the inuence of GA3
level and storage time on titratable acidity and the two-way interac-
tions are presented in Table 2 that signied the interractive inuence of
584
Scientia Horticulturae 225 (2017) 581588
GA3 level and storage period. In our study, the low levels of titratable
acidity in the control fruits in comparison to the coated fruit indicates
that the coating on fruits suspended the ripening mechanism by oering
a semipermeable layer over the fruit (Ali et al., 2010). Titratable acidity
of green chilli coated with GA3 also showed variable results depending
on the dierent coating concentrations. The 2 ppm of GA3 produced
better results as compared 3 and 1 ppm, respectively. At the red skin
colour stage, for both for control and coated fruit, acidity persisted
almost the same whereas it started to decline with the passage of sto-
rage period further. This probably accounted for the reduced rate of
respiratory activity indicates the lower metabolic activities (zden and
Bayindirli, 2002) and delay in utilization of organic acids (Yaman and
Bayindirli, 2002). Our outcome on titratable acidity corresponds with
the previous report on eect of coatings on shelf-life of chilli (zden
and Bayindirli, 2002).
3.3. Ascorbic acid content
Ascorbic acid is one of the vital hydrophilic antioxidant, which
scavenges detrimental free radicals and additional ROS as well as re-
juvenates other antioxidants such as tocopherol to its active form
(Denre et al., 2013). In our study, the quantity of ascorbic acid from
coated green chilli was observed to be greater than control (Table 1).
The 2 ppm concentration of GA3 proved to be the better concentration
where ascorbic acid content of 9.125 0.006 mg/10 g fruit pulp was
detected following 9-days storage that was higher than those of the
control, 1 ppm and 3 ppm GA3 coated fruits. The maximum extension of
shelf-life was observed in 2 ppm GA3 coated chilli that was up to
45 days followed by 36 days in 3 ppm GA3 coated ones. These data
were also supported by the two-way ANOVA where interraction eect
of GA3 concentrations with storage duration observed to be signicant
at P < 0.05 (Table 2). Due to its oxidation, ascorbic acid being a vital
nutrient quality factor is very delicate to degradation (Veltman et al.,
2000) in comparison to other nutrient at the time of food processing
and storage. Ascorbic acid level varies with genotypic variations, pre-
harvest climatic states, ripeness and postharvest treatment processes.
Ascorbic acid content in chilli fruits declined with extending period of
storage. The results of our study were similar to the previous study of
Kumar et al. (2000) in kinnow and Huang et al. (2014) in banana.
Ascorbic acid of vegetables produced maximum value immediately
prior to ripening and later decline owing to the enzymatic activity of
ascorbic acid oxidase (Bhattacharya, 2004). Comparable to the result of
titratable acidity, the 2 ppm concentration of GA3 displayed better re-
sults for ascorbic acid than 3 ppm and 1 ppm in decreasing order.
3.4. Total soluble sugar content
As the days of storage increased in each case of control and coated
chillies, the sugar concentration increased (Table 1). With the progress
of ripening, more micro molecules are produced which was supported
by this study. The highest content of total soluble sugar was found in
2 ppm GA3 treated chillies with a value of 331.067 3.197 mg/g fruit
pulp having maximum shelf-life extension of 45 days. On the other
hand, the control chillies survived only up to 18 days in storage re-
cording a signicantly lower value (282.923 0.285 mg/g). The
above values were shown to be signicant at P < 0.05 (Table 2). The
other treatments also exhibited comparable total sugar content, albeit
lesser shelf-life extension. The present results of total soluble sugar
content gives the conformity of previous studies on use of coatings in
Ziziphus and tomato (Beckles, 2012; Li et al., 2009). The decline of
sugar content throughout storage period is normal, since sugars that are
the major component of the soluble solids content of a produce, are
consumed by respiration and expended for the metabolic activities of
the green chillies (zden and Bayindirli, 2002). However, incompar-
able increase of total soluble sugar in most of the applied conditions
(Table 1) were recorded in the present study. Such result, which
 J. Panigrahi et al.

Table 1
Eect of GA3 coating and storage period on titrable acidity (TA), ascorbic acid content (AAC), total soluble sugar content (TSSC), ferrous ion chelating activity (FICA), reducing power activity (RPA), total phenolic content (TPC), hydroxyl radical
scavenging activity (HRSA), DPPH radical scavenging activity (DPPH RSA), polyphenol oxidase activity (POA), and pectate lyase activity (PLA) in green chilli (Capsicum annuum L.).

GA3 (ppm) Storage TA (% citric acid) AAC (mg/10 g fruit TSS (mg/g fruit pulp) FICA (mm of RPA (absorbance at TPC (mg GAE/g HRSA (%) DPPH RSA (%) POA (U/100 mg PLA (U/g)
(days) pulp) chelated Fe2+) 700 nm) fruit pulp) fruit pulp)

0 0 0.493 0.006 11.483 0.029 75.140 1.059 1.171 0.051 0.275 0.004 0.547 0.017 1.725 0.005 0.761 0.004 0.048 0.002 0.215 0.001
0 9 0.175 0.006 7.793 0.012 155.977 0.351 1.386 0.008 0.280 0.002 0.527 0.019 1.824 0.005 2.121 0.009 0.092 0.001 0.246 0.009
18 0.127 0.006 3.533 0.058 282.923 0.285 1.728 0.006 0.630 0.007 0.327 0.006 1.988 0.002 2.149 0.006 0.084 0.003 0.146 0.003
27
36
45
1 9 0.213 0.006 6.200 0.265 162.007 0.386 1.609 0.012 0.783 0.007 0.552 0.008 1.911 0.009 2.297 0.031 0.098 0.002 0.253 0.001
18 0.157 0.006 4.533 0.058 248.003 0.376 1.749 0.001 1.614 0.066 0354 0.021 2.021 0.009 2.386 0.012 0.095 0.004 0.168 0.008

585
27 0.117 0.006 1.117 0.015 254.030 2.140 1.897 0.005 1.161 0.077 0.246 0.005 2.213 0.006 2.385 0.039 0.086 0.004 0.156 0.003
36
45
2 9 0.307 0.006 9.125 0.006 167.900 0.971 1.847 0.005 1.569 0.003 0.559 0.005 2.080 0.062 2.524 0.050 0.103 0.002 0.254 0.004
18 0.267 0.006 8.257 0.045 250.073 0.991 1.987 0.006 1.629 0.007 0.425 0.006 2.183 0.056 2.479 0.059 0.186 0.004 0.182 0.001
27 0.193 0.006 6.713 0.133 277.953 1.983 2.017 0.006 1.396 0.015 0.311 0.011 2.326 0.014 2.665 0.033 0.124 0.002 0.162 0.001
36 0.153 0.006 5.233 0.058 299.000 4.498 2.220 0.010 1.151 0.052 0.239 0.003 2.462 0.050 2.762 0.009 0.096 0.001 0.126 0.001
45 0.123 0.006 2.800 0.100 331.067 3.197 2.293 0.006 0.918 0.006 0.214 0.005 2.533 0.031 2.864 0.065 0.079 0.006 0.096 0.002
3 9 0.223 0.006 7.967 0.058 165.000 0.677 1.640 0.010 1.425 0.004 0.561 0.002 1.944 0.021 2.341 0.049 0.126 0.003 0.261 0.009
18 0.207 0.006 4.100 0.100 222.967 2.542 1.815 0.005 1.612 0.005 0.403 0.008 2.076 0.004 2.445 0.049 0.142 0.002 0.184 0.002
27 0.167 0.006 3.517 0.076 268.063 2.893 1.947 0.006 1.214 0.009 0.276 0.006 2.350 0.007 2.477 0.025 0.104 0.002 0.136 0.001
36 0.113 0.006 1.433 0.058 280.033 3.002 2.143 0.059 0.907 0.015 0.215 0.005 2.382 0.007 2.529 0.023 0.080 0.010 0.113 0.004
45

The data represent mean standard deviation. The control and the dierent treatments are signicantly dierent (p < 0.05). Absence of data () represent unavailability of information due to postharvest deterioration of fruits.
Scientia Horticulturae 225 (2017) 581588
J. Panigrahi et al. Scientia Horticulturae 225 (2017) 581588

Two-way ANOVA showing individual and interaction eect of the GA3 coating and storage period on titrable acidity (TA), ascorbic acid content (AAC), total soluble sugar content (TSS), ferrous ion chelating activity (FICA), reducing power activity
conicts with the usual decrease, might be attributed to concentrated

1350.779*
11.378
22.274
28.842
3.377*

2486.344

9471.617
385.033*
and ascended levels of soluble sugars owing to the water loss
(RPA), total phenolic content (TPC), hydroxyl radical scavenging activity (HRSA), DPPH radical scavenging activity (DPPH RSA), polyphenol oxidase activity (POA), and pectate lyase activity (PLA) in green chilli (Capsicum annuum L.).

F
throughout the span of cold-storage (zden and Bayindirli, 2002; Koh

F
Mean square et al., 2017).

3.5. Ferrous ion chelating activity

Mean square

1.145E-005
1.110
2.173
2.814
0.329
0.098
RPA

In terms of its ecacy on ferrous ion chelating activity, the 2 ppm

0.028
0.015
0.108
0.004
PLA
concentration of GA3 has shown better results than 3 ppm and 1 ppm in
decreasing order. As the days progresses, though the ferrous ion che-
lating activity increases in each cases but 2 ppm concentration of GA3
2304.078*
3419.543
8087.378
3965.236

proved to be the better among them (Table 1). The 2 ppm concentration

1802.211
2122.514
260.796*
866.572
of GA3 improves the extended rate of ferrous ion chelating activity
F

F
more as compared to the lower and higher concentration of GA3 as well
without coating which was observed to be signicant at P < 0.05
(Table 2). Ferrous ions are one of the most competent pro-oxidants in
Mean square

Mean square

biological systems; they can form highly reactive hydroxyl radicals

9.262E-006
through their interaction with hydrogen peroxide. Ferrozine (ferroin
2.045
4.836
2.371
1.378
0.001
FICA

0.008
0.017
0.020
0.002

compound) develops steady magenta-coloured complexes with ferrous

POA

ion (Fe2+). But the complex formation is hindered and the colour of the
complex fades in the presence of other chelating compounds. The
10181.696*

chelating activity of the coexisting chelator is estimated via the detec-

13989.954
24924.507
18853.997

2778.006*
4948.850
9725.818
8595.204

tion of the rate of colour reduction. In our study, the ferrous ion che-
lating activities are increased with increasing the storage period as
F

agreed by Dinis et al. (1994). Though, the previous study related to use
F

of coating on chilli did not observe the ferrous ion chelating activity.
Nevertheless, the antioxidant studies in chilli suggested of having more
Mean square

Mean square
40904.664
72875.762
55126.443
29769.851

ferrous ion activity in pericarp as compared to seeds (Sim and Sil,

DPPH RSA

2008).
2.924

3.792
7.452
6.586
2.128
0.001
TSS

3.6. Reducing power activity

As observed in the results of other attributes, the 2 ppm con-

35566.480
8322.164*
9315.285

2308.261*
763.511*

3384.024
7120.398
4369.488

centration of GA3 showed better results than 3 ppm and 1 ppm in a

decreasing order (Table 1). Though, 1 ppm concentration of GA3
F

showed the reducing power activity close to that of the 3 ppm, still the
same concentration could not sustain for 45 days as done by 2 ppm GA3
coating (Tables 1, 2). Reducing power is linked with antioxidant ac-
Mean square

Mean square

tivity and serves as an evidence of the antioxidant activity. Compounds

201.543
52.787
47.159

having reducing power revealed that they are electron donors. There-
4.327
0.006

HRSA

2.667
5.611
3.443
1.819
0.001
AAC

fore, they perform as primary and secondary antioxidants. In this

method, the yellow colour solution changes to blue depending on the
reducing power of each compound (Chanda and Dave, 2009). The re-
11417.762

ducing eciency of a compound may function as a key marker of its

1816.937*

1367.518

5403.489
2816.197

718.053*

152.087*
148.860*

strong antioxidant activity. The decreased absorbance of the reaction

complex species decreasing reducing power. The reducing power of
F
F

chillies decreases with expanding storage period. No such activities

shown by earlier study on the use of coating in chilli. Simultaneously,
study in antioxidant activity of Capsicum pericarp, the reducing power
Mean square
Mean square

2.917E-005

activity initially increases and later on decreases in support of the

present study (Sim and Sil, 2008).
0.141
0.074
0.559
0.016
0.000
0.082
0.053
0.333
0.004

TPC
TA

3.7. Total phenolic content

23

15
48
72
23

15
48
72

The results of the study indicated that GA3 coating signicantly

df
df

3
5
3
5

increases the storage life of chillies and its overall quality through af-
fecting the secondary metabolites, antimicrobial and antioxidant
GA3 conc. Storage duration

GA3 conc. Storage duration

properties. The 2 ppm concentration of GA3 displayed better results

than 3 ppm and 1 ppm (Table 1). Throughout cold storage the total
*Signicant at p < 0.05.

phenolic content gradually decreased in all three levels of GA3 treat-

ments and control as well. The two-way ANOVA conrmed the above
Storage duration

Storage duration
Corrected model

Corrected model

results since it was proved to be signicant at P < 0.05 (Table 2).

Phenolic compounds are an important class of secondary metabolites in
GA3 conc.

GA3 conc.

chilli that strongly inuence the quality such as colour, avour and
Source

Source
Table 2

Error

Error
Total

Total

pungency (Chamkha et al., 2003); and display antimicrobial and anti-

oxidant activities (Frankel et al., 1995). The control samples recorded

586
J. Panigrahi et al.
the highest phenolics as they ripened faster as compared to coated
chillies, which conrms the ndings of previous studies (Chitravathi
et al., 2014; Ghasemnezhad et al., 2011). The rise in total phenolic
content is associated with the improvement of antioxidant capacity
(Reyes and Cisneros-Zevallos, 2003). Therefore, in the present report,
the highest total phenolic content in 2 ppm of GA3 coated green chillies
suggests that these fruits sustained higher amounts of antioxidants than
controls or fruits coated with higher (3 ppm) or lower (1 ppm) con-
centration. The low level of total phenolic content or its steep drop
following 18 days of storage in control as well as 1 or 3 ppm GA3 coated
fruit might be owing to the rise in respiration rate that caused the de-
cline of total phenolic content via breakdown of certain phenolic
compounds (Day, 2001). According to Ali et al. (2010) it could also be
due to senescence and degradation of cell structure through storage.
3.8. Hydroxyl radical scavenging activity
It follows the above trend like 2 ppm concentration of GA3 had
better results than 3 ppm and 1 ppm and also signicant at P < 0.05
(Tables 1, 2). As the days progressed, the hydroxyl radical scavenging
activity was increased in all the treatment, though 2 ppm concentration
of GA3 resulted in the superior shelf-life extension up to 45 days as
compared to the control (18 days). The hydroxyl radicals are the most
important active oxygen species causing lipid oxidation and huge bio-
logical damage (Gutteridge, 1984). Hydroxyl radicals were developed
in free solution and get identied by their capability to reduce 2-deoxy-
2-ribose into fractions, which form a pink chromogen on warming with
TBA at low pH. As presented in Table 1 chilli was competent to reduce
hydroxyl radical development at all the GA3 levels and it exerts op-
timum scavenging eects. Hydroxyl radical scavenging activity gradu-
ally increased with the extension of storage period. Such trend of hy-
droxyl radical scavenging activity in the present study supports the
reports of Wang and Gao (2013) on post-harvest fruit quality of
strawberries.
3.9. DPPH radical scavenging activity
It was revealed from the Table 1, the DPPH radical scavenging ac-
tivity gradually increased as the days progressed up to 45 days. The
2 ppm concentration of GA3 showed better DPPH radical values from its
counter surface coating of 3 ppm and 1 ppm. The GA3 coating sig-
nicantly inuenced the DPPH radical scavenging activity (Table 2).
The DPPH radical scavenging activity is established relying on the
power of DPPH, an unchanging free radical, to bleach out the existence
of antioxidants. Incidentally, the antioxidants react with DPPH (stable
free radical with violet colour) to produce a yellow coloured , -di-
phenyl-- picryl hydrazine. Such discolouration of the reaction mixture
can be measured through detection of the absorbance at 517 nm that
signies that radical-scavenging eciency of the antioxidants
(Malterud et al., 1993). A sharp initial increase and a very slow gradual
increase in DPPH activity was observed in tomato and fresh cut pears
that endorse the present study (Ali et al., 2013; Oms-Oliu et al., 2008).
Nevertheless, there are multiple factors e.g. genetic background, en-
vironmental conditions, production methods followed, the day of har-
vest and postharvest storage environments that inuence the anti-
oxidant capacity (Dumas et al., 2003). By and large, a progressive
association has been conrmed among total phenolic content and total
antioxidant capacity (Reyes and Cisneros-Zevallos, 2003). The late in-
crease in antioxidant activity in fruit coated with 2 ppm GA3 could be
associated to the delayed ripening of those fruit in comparison to the
control and 1 or 3 ppm GA3 coated fruit, and it was fairly evident in an
earlier report where the tomato fruits covered by gum Arabic de-
celerated the ripening activity through suspension of the storage-asso-
ciated biochemical and physiological transformations (Ali et al., 2010).
The antioxidant action is very much reliant on fruit ripening courses.
Throughout ripening, the antioxidant activity escalates and this
587
Scientia Horticulturae 225 (2017) 581588
escalation is primarily owing to the alterations into the lipophilic an-
tioxidant movement (Cano et al., 2003).
3.10. Polyphenol oxidase activity
The polyphenol oxidase activity signicantly increases up to
18 days storage period irrespective of the levels of GA3 coating. But
eventually, it started to decrease with the passage of storage period. As
shown in Table 1, its value recorded highest and comparable in the
fruits with 2 ppm and 3 ppm GA3 coating (0.079 0.006 and
0.080 0.010 U/100 mg fruit pulp, respectively) following 45 days of
storage. Contrarily, the untreated fruits showed 0.084 0.003 U/
100 mg but could withstand merely 18 days storage. Polyphenol oxi-
dase is a terminal oxidase which occurs extensively in plants. It cata-
lyses the oxidation of phenolics ensuing in the browning of tissue in
fruits and vegetables (Zhang and Quantick, 1997). The previous study
of chitosan coatings in longan fruit has shown the reduction of poly-
phenol oxidase activity (Jiang and Li, 2001) and the same as the case in
crude plant extracts (Ponce et al., 2008).
3.11. Pectate lyase activity
The pectate lyase activity observed to be inclined during the com-
mencement of storage but later it displayed a decreasing pattern to-
wards the completion of storage phase. The trend of pectate lyase ac-
tivity has been presented in Table 1. As the days progressed, the
ripening process started and the shelf-life of the respective fruits dete-
riorated with increase in the pectate lyase activity. Such activity was
clearly observed initially in control fruits since it was started deterior-
ating immediately after 18 days. The GA3 coated chillies either with
1 ppm or 3 ppm failed to unrestrict the activity of pectate lyase beyond
27 and 36 days, respectively. On the other hand, 2 ppm GA3 coated
chillies displayed an extended storage of 45 days with signicantly
lower pectate lyase activity (Tables 1, 2). Pectate lyase acts as a vital
enzyme during ripening process and involved in softening of fruits and
vegetable tissues. Such restriction in ripening mechanism though con-
trolled pectate lyase activity also observed in green bell peppers and
fresh cut papaya (Conforti and Zinck, 2002; Robles-Snchez et al.,
2013).
4. Conclusion
The results of the present investigation indicated that the green
chillies coated with GA3 showed a signicant delay in the change of
titratable acidity, total phenolic content, decrease ascorbic acid, in-
crease enzymes activity, increase antioxidant activity during the cold
storage compared to uncoated ones. Application of GA3 coating on
green chillies showed expedient impact by delaying the course of ri-
pening. The postharvest treatments by the dierent concentration
(1 ppm, 2 ppm and 3 ppm) of GA3 as a coating extended the storage life
of chillies. From the obtained results, it is indicated that the 2 ppm of
GA3 concentration is better than the 1 ppm or 3 ppm. Hence, 2 ppm of
GA3 concentration can be considered better for storage life of green
chilli. In future, GA3 coating will be a huge potential in fruits and ve-
getables for post-harvest preservation.
Funding
This research did not obtain any specic grant from funding agen-
cies in the public, commercial, or not-for-prot sectors.
Conict of interest
Declared none.
J. Panigrahi et al.
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