Scientia Horticulturae 217 (2017) 156–163

Contents lists available at ScienceDirect

Scientia Horticulturae
journal homepage: www.elsevier.com/locate/scihorti

Quality and biochemical changes of ‘Hindi-Besennara’ mangoes

during shelf life as affected by chitosan, trans-resveratrol and glycine
betaine postharvest dipping
Mohamed A. Awad a,b,∗ , Adel D. Al-Qurashi a , El-Refaey F.A. El-Dengawy c ,
Mohamed I. Elsayed a
a
Department of Arid Land Agriculture, Faculty of Meteorology, Environment and Arid Land Agriculture, King Abdulaziz University, P.O. Box 80208, Jeddah,
Saudi Arabia
b
Pomology Department, Faculty of Agriculture, Mansoura University, El-Mansoura, Egypt
c
Pomology Department, Faculty of Agriculture, Damietta University, 34517, Damietta, Egypt

a r t i c l e i n f o a b s t r a c t

Article history: Effects of chitosan (1%), trans-resveratrol (resveratrol) (1.6 × 10−5 M, 1.6 × 10−4 M and 1.6 × 10−3 M) and
Received 21 October 2016 glycine betaine (GB) (10, 15 and 20 mM) dipping on quality and biochemical changes of ‘Hindi-Besennara’
Received in revised form 24 January 2017 mangoes during ripening at shelf life (SL) (18 ± 2 ◦ C, 60–70% RH) were studied. Resveratrol, especially at
Accepted 25 January 2017
low rate followed by GB, especially at high rate, decreased decay percentage after one and two weeks of
Available online 5 February 2017
SL compared to other treatments. Both compounds at all rates retained higher fruit firmness during SL
and higher titratable acidity (TA) (only after one week of SL). Both GB and resveratrol at all rates retained
Keywords:
higher vitamin C level than control with no effect on total soluble solids (TSS) after two weeks of SL. These
Mango
Chitosan
compounds had no effect on weight loss after one week, but increased it after two weeks of SL compared
Antioxidants to control. Chitosan showed higher weight loss during SL but, retained higher TA and vitamin C, and
Resveratrol lower pH with no significant impact on firmness, TSS and decay compared to control. Chitosan, low and
Glycine betaine medium resveratrol rates, and low and high GB rates maintained higher membrane stability of peel after
Enzymes two weeks of SL compared to control. All treatments showed lower ␣-amylase but, higher peroxidase
(POD) activities in peel than control after two weeks of SL. High resveratrol rate retained higher total
phenols level than control, in contrast to chitosan after two weeks of SL while, total flavonoids was
not affected. Compared to initial, peel free radical scavenging capacity (FRSC) decreased after one week
followed by a sharp increase after two weeks of SL. Chitosan, medium and high resveratrol rates, and
medium GB rate showed higher FRSC than control after one week of SL but, with no differences after two
weeks of SL. In conclusion, both trans-resveratrol and GB treatments retained quality of ‘Hindi-Besennara’
mangoes during SL and being suggested as natural alternatives to synthetic chemicals.
© 2017 Elsevier B.V. All rights reserved.

1. Introduction a temperature below 13 ◦ C (Sivakumar et al., 2011). Synthetic

As climacteric fruit, mangoes (Mangifera indica L.) possess a
relatively short shelf life at ambient conditions due to rapid soften-
ing and the development of several physiological and pathological
disorders (Sivakumar et al., 2011; López-Mora et al., 2013). Also,
mangoes are highly sensitive to chilling injuries when stored at
∗ Corresponding author at: Department of Arid Land Agriculture, Faculty of Mete-
orology, Environment and Arid Land Agriculture, King Abdulaziz University, P.O. Box
80208, Jeddah, Saudi Arabia.
E-mail addresses: mawad@mans.edu.eg, mawad882005@yahoo.com
(M.A. Awad).
chemical preservatives application at pre or postharvest stages is
restricted due to rising consumers concerns on both human health
and the environment. Thus, alternative tools to maintain mangoes
quality during shelf life are critically required. Chitosan, a bioactive
natural edible coat, is widely considered as a promising alterna-
tive to chemical preservatives (Romanazzi et al., 2013; Al-Qurashi
and Awad, 2015). Postharvest dipping of ‘Tainong’ mangoes in
2% chitosan decreased respiration rate, and the loss of firmness,
color change, acidity, ascorbic acid and fruit weight as well as
inhibited diseases progress during storage at 15 ◦ C (Zhu et al.,
2008). Also dipping ‘Nam Dok Mai’ mangoes, previously inocu-
lated with C. gloeosporioides, in chitosan (from 0.5 to 2.0%) delayed
ripening and reduced respiration rate, ethylene production, and

http://dx.doi.org/10.1016/j.scienta.2017.01.043
0304-4238/© 2017 Elsevier B.V. All rights reserved.
 M.A. Awad et al. / Scientia Horticulturae 217 (2017) 156–163
weight loss, ascorbic acid, and acidity and reduced diseases pro-
gression (Jitareerat et al., 2007). However, ‘Tommy Atkins’ mangoes
dipped in 1% chitosan had no effects on fruit ripening, weight loss
and black spot incidence, but inhibited the extension of this dis-
ease during storage at 12 ◦ C and 25 ◦ C (López-Mora et al., 2013).
Resveratrol, its 3-glucopyranoside piceid, and their cis isomers
are natural antioxidant plant phenolics, representing the major
active compound of stilbene phytoalexins that mainly occur in
grapes, berries, and other dietary constituents and are presumed
to be involved in defense system against plant pathogens and
metabolic diseases in human (Adrian et al., 1997; Gonzalez Urena
et al., 2003; Jimenez et al., 2005; Gülçin, 2010; Chen et al., 2016).
Postharvest dipping in trans-resveratrol at 1.6 × 10−4 M maintained
firmness, sensory and nutritional value and reduced water loss dur-
ing storage of apples, grapes and tomatoes compared to control
(Jimenez et al., 2005). Cherukuri (2007) reported that exoge-
nous trans-resveratrol treatment at 1.6 × 10−3 M, 1.6 × 10−4 M
and 1.6 × 10−5 M increased total phenolics, vitamin C and total
carotenoids concentration and antioxidant capacity of Satsuma
mandarins. Postharvest trans-resveratrol dipping at 1.6 × 10−5 M,
1.6 × 10−4 M and 1.6 × 10−3 M reduced decay, increased antioxi-
dant compounds, POD and polyphenoloxidase (PPO) and decreased
polygalacturonase activities of ‘El-Bayadi’ table grapes after cold
storage and shelf life (Awad et al., 2015). Glycine betaine (GB) is a
naturally occurring compatible solute that function as an effective
osmotic stress protectant and stabilizing photosynthetic pigments
and cell membranes in plants (Robinson and Jones, 1986; Genard
et al., 1991; Ashraf and Foolad, 2007; Mansour, 2000; Yang et al.,
2003; Chen and Murata, 2011). Foliar application of GB at 50 and
100 mM/L increased antioxidant enzymes activities and enhanced
photosynthesis of maize under salinity stress (Nawaz and Ashraf,
2009). Postharvest GB dipping of Shredded Iceberg lettuce, at
especially 0.2 mM, maintained sensory quality during cold stor-
age (Hurme et al., 1999). Also, dipping of ‘Zhongnong 8’ cucumbers
in GB at 5, 10, and 15 mM decreased lipoxygenase (LOX) activity
but increased POD and catalase (CAT), restrained malondialdehyde
(MDA) and hydrogen peroxide (H2 O2 ) accumulations, especially at
10 mM during cold storage (Fu-gui et al., 2013). Postharvest GB dip-
ping at 10, 15 and 20 mM increased antioxidant compounds, POD
and PPO activities of ‘El-Bayadi’ table grapes after cold storage and
shelf life (Awad et al., 2015). This study aim to evaluate the response
of ‘Hindi-Besennara’ mangoes to postharvest dipping in chitosan,
resveratrol and GB treatment as an attempt to maintain quality
during SL.
2. Materials and methods
2.1. Plant materials and experimental procedure
This experiment was performed on mature hard-green ‘Hindi-
Besennara’ mangoes collected from a commercial orchard located
in Jizan region (17.4751◦ N, 42.7076◦ E), Kingdom of Saudi Arabia.
Fruit were packed in perforated cardbox (12 fruit of each box, about
3.0–3.5 kg) and transported to the postharvest laboratory at King
Abdulaziz University in Jeddah within about 8 h at 15 ◦ C. Fruit of
uniform size, weight (250–300 g/fruit) and appearance and free of
visual defects were selected for this experiment.
2.2. Fruit treatment
A completely randomized experimental design with three
replicates (20 fruit of each) was established. Fruit of each treat-
ment/replicate were soaked either into water (control), 1% acetic
acid, 1% chitosan (100,000–300,000 MW) (Acros Organic, New
Jersey, USA) dissolved in 1% acetic acid, trans-resveratrol (Baoji
157
Guokang Bio-Technology Co., Ltd., China) solution (1.6 × 10−5 M,
1.6 × 10−4 M and 1.6 × 10−3 M; referring to 0.00365, 0.0365 and
0.365 g/L, respectively) or glycine betaine (Danisco, Finland) solu-
tion (10, 15, 20 mM; refereeing to 1.172, 1.757 and 2.343 g/L,
respectively) for 1 min. A surfactant (Tween 20 at 0.5 mL/L) was
added to all treatments. Following air draying of about 1 h, all
treatments/replicates were weighted and stored at 18 ± 2 ◦ C and
60–70% (RH) in perforated cardboard cartons for 2 weeks. Before
applying the treatments, additional three samples (5 fruit of each)
were randomly collected for initial quality and biochemical anal-
yses as described below. After one and two weeks of shelf life,
weight loss and decay incidence were recorded for each treat-
ment/replicate as described below. Also, samples (5 fruit of each)
from each treatment/replicate were randomly collected for quality
and biochemical analyses. Then, these fruit samples were peeled
and the peel tissue was sliced and mixed. Random part of this peel
was used for electrolyte leakage measurement and the remaining
peel was kept at −80 ◦ C for later total phenols, flavonoids, enzymes
and antioxidant activity analysis. Pulp firmness was measured in
each sample directly following peeling then, the pulp tissue was
sliced and mixed. Random portion of this pulp tissue was directly
used for TSS, TA, pH, and vitamin C determinations.
2.3. Weight loss determination
The total fruit weight loss was calculated on initial weight basis
and expressed in percentage.
2.4. Decay incidence
Decay incidence, due to skin browning, shriveling and diseases,
was recorded and calculated on initial fruit number basis for each
samples and expressed in percentage.
2.5. Firmness, TSS, TA, pH and vitamin C measurements in fruit
pulp
Fruit pulp firmness was measured independently in 5 fruit (two
opposite measurements in the middle of each fruit) per replicate by
a digital basic force gauge, model BFG 50N (Mecmesin, Sterling, VA,
USA) supplemented with a probe of 11 mm diameter and the results
were expressed as Newton. A homogeneous sample was prepared
from these 5 fruit per replicate for measuring TSS content, TA, pH
and vitamin C concentration. TSS content was measured in fruit
pulp juice with a digital refractometer (Pocket Refractometer PAL 3,
ATAGO, Japan) and expressed in percentage. TA was determined in
distilled water diluted fruit juice (1:2) by titrating with 0.1N sodium
hydroxide up to pH 8.2, using automatic titrator (HI 902, HANNA
Instrument, USA) and the results expressed as a percentage of citric
acid. Fruit juice pH was measured by a pH meter (WTW 82382,
Weilheim, Germany). Vitamin C was measured by the oxidation
of ascorbic acid with 2,6-dichlorophenol endophenol dye and the
results expressed as g kg−1 on a fresh weight (FW) basis (Ranganna,
1979).
2.6. Leakage of ions from fruit peel
Leakage of ions was measured in peel disks according to Sairam
et al. (1997) with some modifications and was expressed as mem-
brane stability index percentage (MSI %). Three grams of peel disks
per replicate/treatment was randomly taken and placed in 30 ml
of deionized water at ambient temperature for 4 h in a shaker.
Conductivity before boiling (C1) was measured with an electrical
conductivity digital meter (Orion 150A+, Thermo Electron Corpora-
tion, USA). The same disks were kept in a boiling water bath (100 ◦ C)
158 M.A. Awad et al. / Scientia Horticulturae 217 (2017) 156–163

for 30 min to release all electrolytes, cooled to 22 ± 2 ◦ C with run- Table 1

Decay and weight loss percentage of ‘Hindi-Besennara’ mangoes during shelf life as
ning water, and conductivity after boiling was recorded (C2). MSI
affected by postharvest chitosan, trans-resveratrol and glycine betaine dipping.
was expressed in percentage using the formula: [1 − (C1/C2)] × 100.
Decay (%) Weight loss (%)

2.7. Preparation of methanol extract of fruit peel Initial 0.0 0.0

Treatment (T)
Control 23.6a 9.4f
Two grams of fruit peel (randomly collected from 5 Acetic acid (1%) 24.9a 14.0a
fruit/replicate) were extracted by shaking at 150 rpm for 12 h Chitosan (1%) 24.6a 11.5c
with 20 mL methanol (80%) and filtered through filter paper No. 1. Resveratrol (M)
The filtrate designated as methanol extract that will be used for 1.6 × 10−5 5.3d 9.8ef
1.6 × 10−4 9.4c 9.6f
total phenols, total flavonoids and antioxidant activity estimations.
1.6 × 10−3 7.2cd 10.1ed
Glycine betaine (mM/L)
2.7.1. Estimation of total phenols 10 14.2b 10.6c
15 14.8b 12.4b
Total phenols concentration was measured according to Hoff
20 10.4c 11.2c
and Singleton (1977). Fifty ␮l of the methanol extract was mixed F-test *** ***

with 100 ␮l Folin-Ciocalteu reagent, 850 ␮l of methanol and LSD (0.05) 3.2 0.48
allowed to stand for 5 min at ambient temperature. A 500 ␮l of Shelf life (SL) (weeks)
20% sodium carbonate was added and allowed to react for 30 min. 1 10.9 7.8
2 19.0 14.2
Absorbance was measured at 750 nm. Total phenols was quantified *** ***
F-test
from a calibration curve obtained by measuring the absorbance of T × SL
known concentrations of gallic acid and the results expressed as F-test * ***

g kg−1 FW gallic acid equivalent.
2.7.2. Estimation of total flavonoids
Total flavonoids concentration was determined using a mod-
ified colorimetric method described previously by Zhishen et al.
(1999). Methanol extract or standard solution (250 ␮l) was mixed
with distilled water (1.25 ml) and 5% NaNO2 solution (75 ␮l). After
standing for 6 min, the mixture was combined with 10% AlCl3 solu-
tion (150 ␮l), 1 M NaOH (0.5 ml) and distilled water (275 ␮l) were
added to the mixture 5 min later. The absorbance of the solutions at
510 nm was then measured. Total flavonoids was quantified from a
calibration curve obtained by measuring the absorbance of known
concentrations of catechin and the results expressed as g kg−1 FW
catechin equivalent.
2.7.3. Evaluation of DPPH radical scavenging assay of fruit peel
Free radical scavenging capacity (FRSC) of methanol extract of
fruit peel was determined using the 1,1-diphenyl-2-picrylhydrazyl
(DPPH) method (Ao et al., 2008). A methanol extract (0.1 ml)
was added to 0.9 ml of freshly prepared DPPH methanol solution
(0.1 mM). An equal amount of methanol was used as a control.
After incubation for 30 min at room temperature in the dark,
the absorbance (Abs) was measured at 517 nm using a spec-
trophotometer. Activity of scavenging (%) was calculated using the
following formula:
 (Abs control − Abs sample) 
DPPH radical scavenging % =
Abs control
×100
Means within each column followed by the same letter are not significantly different
at level P ≤ 0.05.
*
Significant at P ≤ 0.05.
***
Significant at P ≤ 0.001.
2.8.2. ˛-Amylase assay
␣-Amylase (EC 3.2.1.1) activity was assayed by determining the
liberated reducing end products using maltose as a standard (Miller,
1959). The reaction mixture (0.5 ml) containing 5 mg substrate,
0.25 ml of 0.2 M sodium acetate buffer pH 5.5 and a suitable amount
of crude extract. Assays were carried out at 37 ◦ C for 1 h. Then 0.5 ml
dinitrosalicylic acid reagent was added to each tube and heated in
a boiling water bath for 10 min. After cooling to room tempera-
ture, the absorbance was measured at 560 nm. Starch was used as
a substrate. One unit of enzyme activity was defined as the amount
of enzyme which liberated 1 ␮M of reducing sugar per min under
standard assay conditions.
2.8.3. Peroxidase assay
Peroxidase (EC 1.11.1.7) activity was assayed according to
Miranda et al. (1995). The reaction mixture containing in one
ml: 0.008 ml of 0.97 M H2 O2 , 0.08 ml of 0.5 M guaiacol, 0.25 ml of
0.2 M sodium acetate buffer, pH 5.5 and least amount of enzyme
preparation. The change in absorbance at 470 nm due to guaiacol
oxidation was followed for 1 min using a spectrophotometer. One
unit of enzyme activity was defined as the amount of enzyme that
increases the O.D. 1.0 per min under standard assay conditions.
2.9. Statistical analysis

The inhibition concentration (IC50 ) was defined as ␮g phenolics
of the test sample that decreases 50% of initial radical. The IC50
values were calculated from the dose responses curves.
2.8. Enzymes measurements of fruit peel
The data were statistically analyzed as a completely randomized
design with three replicates by analysis of variance (ANOVA) using
the statistical package software SAS (SAS Institute Inc., 2000, Cary,
NC, USA). Comparisons between means were made by F-test and
the least significant differences (LSD) at P ≤ 5%.

2.8.1. Crude extract preparation 3. Results

One gram of fruit peel (randomly collected from 5
fruit/replicate) was homogenized with 20 mM Tris–HCl buffer,
pH 7.2 using homogenizer. The homogenate was centrifuged at
10,000 rpm for 10 min at 4 ◦ C. The supernatant was designed
as crude extract and stored at −20 ◦ C for both ␣-amylase and
peroxidase assay.
There were significant interaction effects between treatment
and SL on decay and weight loss percentage (Tables 1 and 2). Decay
percentage increased during SL in all treatments, except for the
medium and high rates of resveratrol. In this respect, after one and
two weeks of shelf life, decay percentage was lower in resveratrol
 M.A. Awad et al. / Scientia Horticulturae 217 (2017) 156–163 159

Table 2
The interaction effect between treatment and shelf life on decay and weight loss percentage of ‘Hindi-Besennara’ mangoes as affected by postharvest chitosan, trans-resveratrol
and glycine betaine dipping.

Treatment Shelf life (weeks)

Decay (%) Weight loss (%)

1 2 1 2

Control 17.7b 29.4a 7.3ij 11.6f

Acetic acid (1%) 19.5b 30.3a 7.4ij 20.6a
Chitosan (1%) 18.9b 30.3a 8.7h 14.3cd
Resveratrol (M)
1.6 × 10−5 2.3f 8.2de 7.2j 12.5e
1.6 × 10−4 7.8de 11.1cd 6.9j 12.2ef
1.6 × 10−3 5.9ef 8.6de 7.4ij 12.8e
Glycine betaine (mM/L)
10 9.2de 19.2b 7.5ij 13.7d
15 11.5cd 18.0b 9.4g 15.3b
20 5.2ef 15.5bc 7.9i 14.4c

For each parameter, means within and between columns followed by the same letter are not significantly different at level P ≤ 0.05.

Table 3
Quality characteristics and peel membrane stability index (MSI) of ‘Hindi-Besennara’ mangoes during shelf life as affected by postharvest chitosan, trans-resveratrol and
glycine betaine dipping.

Firmness TSS Acidity pH Vitamin C MSI

(N) (%) (%) (g kg−1 ) (Index)

Initial 6.90 12.8 1.35 3.42 0.45 20.0

Treatment (T)
Control 3.85ed 17.3bcd 0.23c 5.3a 0.31c 8.8e
Acetic acid (1%) 3.41e 16.0d 0.28bc 4.9bc 0.46a 11.4cd
Chitosan (1%) 4.14d 16.7bcd 0.42ab 4.8bc 0.45a 19.3a
Resveratrol (M)
1.6 × 10−5 5.44ab 16.0d 0.37ab 4.8bc 0.38b 17.9a
1.6 × 10−4 5.48ab 17.6abc 0.47a 4.9b 0.43ab 15.1b
1.6 × 10−3 4.74c 19.0a 0.47a 4.9b 0.46a 9.7de
Glycine betaine (mM/L)
10 5.87a 17.2bcd 0.42ab 4.8bc 0.45a 12.8c
15 5.55ab 16.5cd 0.42ab 4.7bc 0.47a 9.2de
20 5.11bc 18.1ab 0.50a 4.7c 0.45a 8.8e
*** *** *** *** *** ***
F-test
LSD (0.05) 0.45 1.48 0.14 0.24 6.1 2.2
Shelf life (SL) (weeks)
1 5.15a 15.5b 0.56a 4.5b 0.42a 15.3a
2 4.54b 18.2a 0.24b 5.2a 0.44a 11.2b
*** *** *** *** ***
F-test NS
T × SL
*** *** *** ***
F-test NS NS

Means within each column followed by the same letter are not significantly different at level P ≤ 0.05. NS, not significant.
***
Significant at P ≤ 0.001.

Table 4
The interaction effect between treatment and shelf life on acidity (%), pH, vitamin C (g kg−1 ) and peel membrane stability index (MSI) of ‘Hindi-Besennara’ mangoes as affected
by postharvest chitosan, trans-resveratrol and glycine betaine dipping.

Treatment Shelf life (weeks)

Acidity (%) pH Vitamin C MSI

1 2 1 2 1 2 1 2

Control 0.20bc 0.25bc 5.4ab 5.2abc 0.33fg 0.30g 9.4efg 8.2fg

Acetic acid (1%) 0.38bc 0.19c 4.4f 5.4ab 0.50ab 0.42bcde 13.0d 9.7efg
Chitosan (1%) 0.63a 0.22bc 4.3f 5.4a 0.37defg 0.53a 21.6a 16.9b
Resveratrol (M)
1.6 × 10−5 0.39b 0.35bc 4.9cd 4.7de 0.36efg 0.41cdef 23.9a 11.8de
1.6 × 10−4 0.73a 0.22bc 4.7de 5.1bcd 0.39cdef 0.46abc 16.6bc 13.5d
1.6 × 10−3 0.70a 0.24bc 4.4ef 5.4ab 0.50ab 0.43bcde 10.9def 8.4fg
Glycine betaine (mM/L)
10 0.64a 0.19c 4.3f 5.2abc 0.40cdef 0.50ab 13.7cd 11.9de
15 0.62a 0.23bc 4.2f 5.2abc 0.45abcd 0.50ab 10.8def 7.5g
20 0.75a 0.25bc 4.2f 5.2abc 0.46abc 0.45abc 17.2b 13.0d

For each parameter, means within and between columns followed by the same letter are not significantly different at level P ≤ 0.05.

and GB treatments at all rates than other treatments (Table 2). After After one week of SL, there were no significant differences among
one and two weeks of SL, the low rate of resveratrol and the high the applied treatments, except for chitosan and the medium rate
rate of GB showed the lowest decay percentage compared to other of GB that showed higher weight loss percentage than other treat-
rates. Weight loss percentage increased during SL in all treatments. ments (Table 2). However, after two weeks of SL, all the applied
160 M.A. Awad et al. / Scientia Horticulturae 217 (2017) 156–163

treatments showed higher weight loss percentage than control, Table 5

Total phenols and flavonoids concentration and free radical scavenging capacity
except for the medium rate of resveratrol. In this respect, resver-
(FRSC) of peel of ‘Hindi-Besennara’ mangoes during shelf life as affected by posthar-
atrol at all rates gave lower weight loss percentage than GB at all vest chitosan, trans-resveratrol and glycine betaine dipping.
rates. Firmness decreased during SL, showed lower values than ini-
Phenols Flavonoids FRSC
tial, and was significantly higher at resveratrol and GB at all rates
(g kg−1 ) (g kg−1 ) (DPPH IC50 value)
than other treatments (Table 3). In this respect, the low and medium
rates of resveratrol showed higher firmness than the high rate. Also, Initial 20.5 1.21 9.5
Treatment (T)
the low rate of GB gave higher firmness than the high rate. However,
Control 22.4cd 2.00ab 17.0bc
there were no significant differences in firmness among chitosan, Acetic acid (1%) 26.5abc 1.51de 20.6ab
acetic acid and control treatments. TSS content increased during Chitosan (1%) 17.7e 1.72bcde 13.1cd
SL, showed higher values than initial, and was higher at the high Resveratrol (M)
1.6 × 10−5 24.6bcd 1.44e 17.4b
rate of resveratrol than control (Table 3). There were no signifi-
1.6 × 10−4 21.1de 1.85abcd 11.7d
cant interaction effect between treatment and SL on fruit firmness 1.6 × 10−3 22.7cd 2.12a 9.9d
and TSS content (Table 3). There were significant interaction effects Glycine betaine (mM/L)
between treatment and shelf life on TA, pH, vitamin C and MSI 10 29.3a 1.65cde 17.0bc
(Tables 3 and 4). TA concentration showed much lower values than 15 27.7ab 1.89abc 11.8d
20 23.7bcd 1.88abc 22.7a
initial and decreased during SL in all treatments, except for con- *** *** ***
F-test
trol, acetic acid and the low rate of resveratrol (Tables 3 and 4). In LSD (0.05) 4.4 0.35 4.0
this respect, after one week of SL chitosan, resveratrol at medium Shelf life (SL) (weeks)
and high rates and GB at all rates showed higher TA concentra- 1 26.5a 1.66b 27.2a
2 21.4b 1.91a 4.2b
tion than acetic acid and control treatments (Table 4). However, *** *** ***
F-test
after two weeks of shelf life there were no significant differences T × SL
among treatments. pH showed much higher values than initial and F-test *** ** ***

increased during SL in all treatments, except for control and the
low and medium rates of resveratrol (Tables 3 and 4). After one
week of SL, pH was lower in all treatments than control while,
after two weeks of SL, pH was lower at the low rate of resveratrol
than other treatments, except for the medium rate of resveratrol
(Table 4). Vitamin C concentration showed similar values to ini-
tial and increased during SL only in chitosan and the low rate of
GB treatments (Tables 3 and 4). After one week of SL, vitamin C
concentration was higher in acetic acid, high rate of resveratrol,
and medium and high rates of GB than other treatments (Table 4).
However, after two weeks of shelf life, all treatments gave higher
vitamin C concentration than control. In this respect, there were no
significant differences in vitamin C concentration among the differ-
ent rates of resveratrol and GB. MSI showed lower values than initial
and decreased during SL in all treatments, except for control and
the low rate of GB (Tables 3 and 4). After one week of SL, the applied
treatments maintained higher MSI than control, except for the high
rate of resveratrol and the medium rate of GB (Table 4). However,
after two weeks of SL, chitosan, the low and medium rates of resver-
atrol and the low and high rates of GB showed higher MSI than
other treatments including control. There were significant interac-
tion effect between treatment and shelf life on total phenols and
flavonoids concentrations and FRSC values (Tables 5 and 6). Total
phenols concentration decreased during SL in acetic acid, chitosan,
Means within each column followed by the same letter are not significantly different
at level P ≤ 0.05.
**
Significant at P ≤ 0.01.
***
Significant at P ≤ 0.001.
low rate of resveratrol, and low and medium rates of GB treatments,
while it did not change in the other treatments (Table 6). After one
week of SL, acetic acid, and the low and medium rates of GB showed
higher total phenols concentration than other treatments. How-
ever, after two weeks of SL, the high rate of resveratrol gave higher
total phenols concentration than control, in contrast to chitosan
treatment. Total flavonoids concentration increased during SL only
in acetic acid and the low rate of GB while, it did not significantly
change in the other treatments (Table 6). After one week of SL, total
flavonoids concentration was lower in acetic acid, and the low rate
of resveratrol and GB than control. However, after two weeks of SL,
there were no significant differences in total flavonoids concentra-
tion among the treatments. Compared to initial, FRSC of peel greatly
decreased (higher DPPH IC50 values) after one week followed by
a sharp increase (lower DPPH IC50 values) after two weeks of SL
(Tables 5 and 6). After one week of SL, chitosan, the medium and
high rates of resveratrol and the medium rate of GB showed higher
FRSC than other treatments (Table 6). However, after two weeks of

Table 6
The interaction effect between treatment and shelf life on total phenols and flavonoids concentration (g kg−1 ) and free radical scavenging capacity (FRSC) (DPPH IC50 value)
in peel of ‘Hindi-Besennara’ mangoes as affected by postharvest chitosan, trans-resveratrol and glycine betaine dipping.

Treatment Shelf life (weeks)

Phenols Flavonoids FRSC

1 2 1 2 1 2

Control 24.5cde 20.2def 2.1a 1.8abc 31.0bc 2.9f

Acetic acid (1%) 34.2ab 18.8efg 1.0e 2.0ab 35.8b 5.2f
Chitosan (1%) 21.7def 13.6g 1.8abc 1.6bcd 22.7d 3.4f
Resveratrol (M)
1.6 × 10−5 30.4abc 18.8efg 1.4cde 1.4cde 31.3bc 3.4f
1.6 × 10−4 21.6def 20.6def 1.7abcd 2.0ab 19.7de 3.7f
1.6 × 10−3 16.1fg 29.3bc 2.1ab 2.1a 14.5e 5.3f
Glycine betaine (mM/L)
10 33.3ab 25.2cd 1.2de 2.0ab 28.9c 5.1f
15 35.8a 19.7defg 1.6abcd 2.1ab 18.9de 4.7f
20 21.4def 25.9cd 1.8abc 1.9ab 41.9a 3.5f

For each parameter, means within and between columns followed by the same letter are not significantly different at level P ≤ 0.05.
 M.A. Awad et al. / Scientia Horticulturae 217 (2017) 156–163 161

Table 7 4. Discussion
␣-amylase and peroxidase activity of ‘Hindi-Besennara’ mangoes peel during shelf
life as affected by postharvest chitosan, trans-resveratrol and glycine betaine
dipping. As a climacteric fruit, mango fruit show a relatively high rate of
physiological activity including high respiration rate and ethylene
␣-amylase Peroxidase
production following harvest that shorten the SL. Due to raising
(U min g FW) (U min g FW)
consumers concerns against the use of synthetic chemicals, several
Initial 0.45 0.90 natural compounds are currently evaluated for their effectiveness
Treatment (T)
on delaying ripening and maintaining fruit quality during SL. In
Control 0.78a 1.13f
Acetic acid (1%) 0.58e 1.58bc this regard, the current study evaluated the response of ‘Hindi-
Chitosan (1%) 0.62d 1.19f Besennara’ mangoes to postharvest dipping in chitosan, as a natural
Resveratrol (M) edible coating, resveratrol, as a natural antioxidant compound, and
1.6 × 10−5 0.73b 1.43de
GB, as a natural compatible solute that function as an osmotic stress
1.6 × 10−4 0.68c 1.33e
1.6 × 10−3 0.62d 1.64b protectant and stabilizing photosynthetic pigments and cell mem-
Glycine betaine (mM/L) brane in plants. Our results showed that resveratrol, especially at
10 0.68c 0.99g low rate followed by GB, especially at high rate treatments, signif-
15 0.68c 1.50cd icantly decreased decay percentage after one and two weeks of SL
20 0.70c 2.11a
*** *** compared to control and other treatments (Tables 1 and 2). More-
F-test
LSD (0.05) 0.025 0.10 over, both compounds at all rates retained higher fruit firmness
Shelf life (SL) (weeks) during SL and higher TA (only after one week of SL). Also, after two
1 0.56b 1.01b weeks of SL, both GB and resveratrol at all rates retained higher level
2 0.79a 1.85a
*** ***
of vitamin C than control with no negative impact on TSS content
F-test
T × SL
(Tables 3 and 4). However, these compounds showed no signifi-
F-test *** *** cant effect on weight loss after one week, but increased it after two
Means within each column followed by the same letter are not significantly different
weeks of SL compared to control. These results partially confirm
at level P ≤ 0.05. those of Jimenez et al. (2005) in which trans-resveratrol especially
***
Significant at P ≤ 0.001. at 1.6 × 10−4 M reduced water loss, maintained turgidity and firm-
ness of apples, grapes and tomatoes during storage compared to
control with no negative impact on other fruit quality. Such effects
Table 8 were attributed to two distinct mechanisms namely the fungicidal
The interaction effect between treatment and shelf life on ␣-amylase and peroxidase and the impermeabilizer properties of trans-resveratrol, as sug-
activity (U min g FW) of ‘Hindi-Besennara’ mangoes peel as affected by postharvest gested by Jimenez et al. (2005). Accordingly, trans-resveratrol may
chitosan, trans-resveratrol and glycine betaine dipping. form a coat on fruit surface that decrease water loss and retain
Treatment Shelf life (weeks) higher fruit firmness than control. trans-resveratrol dipping of
‘Crimson’ table grapes controlled decay, but showed no synergistic
␣-amylase Peroxidase
effect when applied in combination with chitosan. trans-resveratrol
1 2 1 2 proved broad antifungal activity especially against Botrytis cinerea
Control 0.66e 0.89a 0.81k 1.4fg (Gonzalez Urena et al., 2003; Jimenez et al., 2005). Also, Awad
Acetic acid (1%) 0.46i 0.70e 1.3gh 1.8c et al. (2015) reported that both trans-resveratrol and GB treatments
Chitosan (1%) 0.49hi 0.75d 1.1i 1.3h retained higher firmness but had no effect on weight loss, TSS, TA
Resveratrol (M)
1.6 × 10−5 0.68e 0.78cd 0.97ij 1.9c
and pH after cold storage and SL of ‘El-Bayadi’ table grapes. The
1.6 × 10−4 0.53g 0.82b 0.90jk 1.7cd induction of natural defense system that include trans-resveratrol
1.6 × 10−3 0.46i 0.77d 1.0i 2.2b accumulation in ‘Cardinal’ grapes tissue by pre-storage carbon
Glycine betaine (mM/L) dioxide treatment was critical in controlling fungus decay dur-
10 0.61f 0.75d 0.43l 1.5ef
ing storage and shelf life. The observed positive effects of GB on
15 0.53gh 0.83b 0.90jk 2.1b
20 0.58f 0.82bc 1.6de 2.6a decay and other quality parameters of ‘Hindi-Besennara’ mangoes
(Tables 1–4) might be related to its well known general properties
For each parameter, means within and between columns followed by the same letter
are not significantly different at level P ≤ 0.05.
as a natural compatible solute that function as a photosynthetic
pigment and membrane stabilizing agent and osmoregulator in
many plant species (Robinson and Jones, 1986; Genard et al., 1991;
Ashraf and Foolad, 2007; Mansour, 2000; Yang et al., 2003; Chen
and Murata, 2011). However, the exact mechanism of especially GB
SL there were no significant differences in FRSC among treatments. on fruit physiology needs further investigations. Zhang et al. (2016)
There were significant interaction effect between treatment and reported that betaine was not able to scavenge free radicals (as
shelf life on ␣-amylase and peroxidase activities (Tables 7 and 8 measured by classical chemical assays such as DPPH, ABTS, FRAP)
). ␣-Amylase activity increased during SL and showed higher or enhance the enzymatic antioxidant system in human hepato-
values than initial (Tables 7 and 8). After one week of SL, both the cellular carcinoma (HepG2) cells. However, such classical assays
low rate of resveratrol and control showed higher ␣-amylase activ- did not always reflect the antioxidant activity, since a compound
ity than other treatments (Table 8). However, after two weeks of SL, without free radical scavenging ability may still possess antioxidant
all the treatments showed lower ␣-amylase activity than control. activity in an organism. Accordingly, Zhang et al. (2016) suggested
POD activity increased during SL in all treatments (Table 8). After that betaine exert its antioxidant activity via two mechanisms.
one week of SL, POD activity was lower in control, medium rate One mechanism involves scavenging reactive oxygen species (ROS)
of resveratrol, and low and medium rates of GB than other treat- in cells via up-regulation of endogenous non-enzymatic antioxi-
ments (Table 8). However, after two weeks of SL, all the treatments dant defense such as increasing the levels of S-adenosylmethionine
showed higher peroxidase activity than control, in contrast to chi- and methionine via the methionine-homocysteine cycle. The other
tosan treatment. After both one and two weeks of shelf life, the high inhibits ROS generation by forming a protective membrane with an
rate of GB treatment showed the highest POD activity. electronegative outer surface around cells, thereby preventing the
162 M.A. Awad et al. / Scientia Horticulturae 217 (2017) 156–163
contact of free radicals with the cytomembrane. It is well known
that weight loss during SL is mainly due to loss of water by tran-
spiration through fruit peel and also to the respiration process
(Jitareerat et al., 2007; Zhu et al., 2008; Jagadeesh et al., 2015). Chi-
tosan as an edible coating is expected to reduce transpiration and
thus decrease weight loss during SL. However, in our experiment,
chitosan showed higher weight loss after one and two weeks of
SL but, retained higher TA and vitamin C, and lower pH with no
significant impact on firmness, TSS and decay of fruit compared to
control (Tables 1–4). These results partially confirm those of López-
Mora et al. (2013) in which 1% chitosan dipping of ‘Tommy Atkins’
mangoes had no effects on fruit ripening, weight loss and black spot
incidence, but inhibited the extension of this disease during stor-
age at 12 ◦ C and 25 ◦ C. However, our results partially contradict with
those of Jitareerat et al. (2007), Zhu et al. (2008) and Jagadeesh et al.
(2015) on mangoes where edible coatings such as chitosan and gum
Arabic delayed ripening and reduced the loss in weight, vitamin C
and TA. Such contradictions are possibly attributed to the degree
of deacetylation (DD), molecular weight and concentration of the
used chitosan that affect the quality and homogeneity of the result-
ing coating. The weight loss of bananas coated with 70% DD chitosan
was higher than those coated with 80% DD chitosan (Suseno et al.,
2014). They also found that increasing chitosan concentration from
1 to 2% (w/w) increased the barrier to the moisture loss and
reduced weight loss during SL, but that was not true for vitamin
C. Mangoes treated with either 100 ppm laboratory-produced chi-
tosan or 500 ppm Sigma-produced chitosan delayed fruit ripening
and effectively decreased the incidence of postharvest stem-end
rot disease (caused by D. natalensis) (Baclayon and Calibo, 2013).
Our results showed that chitosan, resveratrol at low and medium
rates, and GB at low and high rates significantly maintained higher
membrane stability index (MSI) of peel tissues after two weeks of SL
(Table 4) compared to control. Fruit ripening and senescence is pos-
sibly an oxidative process in which the transition from mature stage
into ripening/senescence stage is accompanied by a progressive
shift toward an oxidative state (Goulao and Oliveira, 2008). Accord-
ingly, excessive ROS production could participate in the oxidation
of lipids and proteins of cell membrane that are involved in mango
ripening. Indeed a steady decrease in membrane stability index, as
measured by the leakage of ions, was observed upon the progres-
sion of fruit ripening (Tables 3 and 4), indicates a gradual loss of
membrane’s stability due to changes occurring in the biochemi-
cal and biophysical properties of cell membranes. During ripening,
the burst in ROS was combined with a surge in the expression of
genes that encode enzymes involved in the generation of antioxi-
dant systems in fruit skin and flesh such as POD, PPO and catalase
to enhance resistance against pathogens were reported (Ali et al.,
2011; Zhang et al., 2013). Our results showed that all the applied
treatments maintained higher vitamin C concentration than control
after two weeks of SL. Also, all treatments showed lower activity
of the hydrolytic enzyme ␣-amylase but, higher the antioxidant
enzyme peroxidase in peel tissues than control after two weeks of
SL (Table 8). These results might indicate that these compounds,
especially resveratrol and GB, enhanced the antioxidant network
of fruit, providing more efficient control of metabolic free radicals
level, thus maintain cell membranes integrity of peel and retain
higher flesh firmness. However, after two weeks of SL, high resver-
atrol rate retained higher total phenols concentration than control,
in contrast to chitosan while, total flavonoids concentration was not
affected (Table 6). After one week of SL, chitosan, the medium and
high rates of resveratrol and the medium rate of GB showed higher
FRSC than control (Table 6). However, there were no differences in
FRSC among all treatments after two weeks of SL. These results par-
tially confirm those of Cherukuri (2007) in which trans-resveratrol
dipping at different concentration (1.6 × 10−3 M, 1.6 × 10−4 M and
1.6 × 10−5 M) on ‘Satsuma’ mandarin retained higher total phenols,
vitamin C and total carotenoids concentration and antioxidant
capacity during 12 weeks of cold storage but, with no effect on total
flavonoids concentration. Also, our results are in partial agreement
with those of Fu-gui et al. (2013) in which dipping of ‘Zhongnong 8’
cucumbers in GB at 5, 10, and 15 mM decreased lipoxygenase (LOX)
activity but increased POD and catalase (CAT), restrained malon-
dialdehyde (MDA) and hydrogen peroxide (H2 O2 ) accumulations,
especially at 10 mM/L during cold storage. Moreover, postharvest
GB dipping at 10, 15 and 20 mM increased antioxidant compounds,
POD and PPO activities of ‘El-Bayadi’ table grapes after cold stor-
age and shelf life (Awad et al., 2015). Compared to initial, FRSC of
fruit peel greatly decreased (higher DPPH IC50 values) after one
week followed by a sharp increase (lower DPPH IC50 values) after
two weeks of SL (Tables 5 and 6). The increase in the antioxi-
dant capacity (lower IC50 values) during shelf life confirm those
of Kondo et al. (2005) where DPPH-radical scavenging activity of
‘Choke anan’ mangoes peel increased during 10 days of storage
at 6 and 12 ◦ C. The decrease in total phenols and the relative sta-
bility of total flavonoids levels with the increase in FRSC (lower
DPPH IC50 values) of peel during SL (Tables 5 and 6) might suggest
qualitative changes in phenolic classes toward higher antioxidant
potential. In other fleshy climacteric fruit, Fernando et al. (2014)
found no significant correlation between the total phenols con-
centration and antioxidant activity measured by DPPH and FRAP,
except for vitamin C level and FRAP in ‘Khai’ banana pulp. However,
Sulaiman et al. (2011) obtained only minor to moderate correlation
between phenolic concentration and antioxidant activities in nine
Malaysian banana cultivars. These results imply that antioxidant
compounds other than phenolics and vitamin C such as carotenoids
and xanthophylls might also be involved. It was reported that the
antioxidant capacity of phenols possibly has a concentration sat-
uration limit above which the activity could not increase further
with the concentration (Dani et al., 2012). Also, parallel different
assays should be used to investigate the principles of antioxi-
dant/oxidation activity of a certain horticultural commodity (Ciz
et al., 2010). In conclusion, both postharvest trans-resveratrol and
GB treatments retained quality of ‘Hindi-Besennara’ mangoes dur-
ing ripening at shelf life conditions and being suggested as natural
alternatives to synthetic chemicals.
Acknowledgments
This project was funded by the Deanship of Scientific Research
(DSR) at King Abdulaziz University of Saudia Arabia, Jeddah, under
grant G-37-155-1437. The authors, therefore, acknowledge with
thanks DSR for technical and financial support. Also, we would like
to thank Nageeb Al-Masoudi, M.Sc., and Nour Gamal, B.Sc. at the
Arid Land Agriculture Department, Faculty of Meteorology, Envi-
ronment and Arid Land Agriculture, King Abdulaziz University, for
their indispensable technical support.
References
Adrian, M., Jeandet, P., Veneau, J., Weston, L.A., Bessis, R., 1997. Biological activity
of resveratrol, a stilbenic compound from grapevines, against Botrytis cinerea,
the causal agent for gray mold. J. Chem. Ecol. 23, 1689–1702.
Ali, M.B., Howard, S., Chen, S., Wang, Y., Yu, O., Kovacs, L.G., Qiu, W., 2011. Berry
skin development in Norton grape: distinct patterns of transcriptional
regulation and flavonoid biosynthesis. BMC Plant Biol. 11, 7–29.
Al-Qurashi, A.D., Awad, M.A., 2015. Postharvest chitosan treatment affects quality,
antioxidant capacity, antioxidant compounds and enzymes activities of
‘El-Bayadi’ table grapes after storage. Sci. Hortic. 197, 392–398.
Ao, C., Li, A., Elzaawely, A.A., Xuan, T.D., Tawata, S., 2008. Evaluation of antioxidant
and antibacterial activities of Ficus microcarpa L. fil. extract. Food Control 19,
940–948.
Ashraf, M., Foolad, M.R., 2007. Roles of glycine betaine and proline in improving
plant abiotic stress resistance. Environ. Exp. Bot. 59, 206–216.
Awad, M.A., Al-Qurashi, A.D., Mohamed, S.A., 2015. Postharvest trans-resveratrol
and glycine betaine treatments affect quality, antioxidant capacity, antioxidant
 M.A. Awad et al. / Scientia Horticulturae 217 (2017) 156–163
compounds and enzymes activities of ‘El-Bayadi’ table grapes after storage and
shelf life. Sci. Hortic. 197, 350–356.
Baclayon, D.P., Calibo, C.L., 2013. Effects of chitosan isolated from crab exoskeleton
on postharvest stem-end rot disease and on the quality of mango fruit. Annal.
Tropic. Resea. 35, 23–34.
Chen, T.H., Murata, N., 2011. Glycine betaine protects plants against abiotic stress:
mechanisms and biotechnological applications. Plant Cell Environ. 34, 1–20.
Chen, M.-L., Yi, L., Zhang, Y., Zhou, X., Ran, L., Yang, J., Zhu, J.-D., Zhang, Q.-Y., Mi,
M.-T., 2016. Resveratrol attenuates trimethylamine-N-oxide (TMAO)-induced
atherosclerosis by regulating TMAO synthesis and bile acid metabolism via
remodeling of the gut microbiota. mBio 7, e02210–e2215.
Cherukuri, K., (M.Sc. thesis) 2007. Effect of trans-resveratrol on shelf life and
bioactive compounds in Satsuma mandarin. Auburn, Alabama, USA, 96 pp.
Ciz, M., Cizova, H., Denev, P., Kratchanova, M., Slavov, A., Lojek, A., 2010. Different
methods for control and comparison of the antioxidant properties of
vegetables. Food Control 21, 518–523.
Dani, C., Oliboni, L.S., Pra, D., Bonatto, D., Santos, C.E., Yoneama, M.L., Dias, J.F.,
Salvador, M., Henriques, J.A.P., 2012. Mineral content is related to antioxidant
and antimutagenic properties of grape juice. Genet. Mol. Res. 11,
3154–3163.
Fernando, H.R.P., Srilaong, V., Pongprasert, N., Boonyaritthongchai, P., Jitareerat, P.,
2014. Changes in antioxidant properties and chemical composition during
ripening in banana variety ‘Hom Thong’ (AAA group) and ‘Khai’ (AA group). Int.
Food Res. J. 21, 749–754.
Fu-gui, W., Tao, H., Hai-ying, Z., Li, X., 2013. Effect of exogenous glycine betaine on
oxidative metabolism in cucumber during low-temperature storage. Food Sci.
34, 313–316.
Genard, H., Le Saos, J., Hillard, J., Tremolieres, A., Boucaud, J., 1991. Effect of salinity
on lipid composition, glycine betaine content and photosynthetic activity in
chloroplasts of Suaeda maritime. Plant Physiol. Biochem. 29, 421–427.
Gonzalez Urena, A.G., Orea, J.M., Montero, C., Jimenez, J.B., Gonzalez, J.L., Sanchez,
A., Dorado, M., 2003. Improving postharvest resistance in fruits by external
application of trans-resveratrol. J. Agric. Food Chem. 51, 82–89.
Goulao, L.F., Oliveira, C.M., 2008. Cell wall modifications during fruit ripening:
when a fruit is not the fruit. Trends Food Sci. Technol. 19, 4–25.
Gülçin, İ., 2010. Antioxidant properties of resveratrol: a structure–activity insight.
Innov. Food Sci. Emerg. Technol. 11, 210–218.
Hoff, J.F., Singleton, K.I., 1977. A method for determination of tannin in foods by
means of immobilized enzymes. J. Food Sci. 42, 1566–1569.
Hurme, E.U., Kinnunen, A., Heiniö, R.L., Ahvenainen, R., Jokinen, K., 1999. The
storage life of packed shredded Iceberg lettuce dipped in glycine betaine
solutions. J. Food Prot. 62, 363–367.
Jagadeesh, N.S.L., Suresha, G.J., Swamy, G.S.K., 2015. Influence of eco-friendly
post-harvest treatments on pulp chroma and hue on mango cv. Alphonso
fruits. The Bioscan 10, 29–32.
Jimenez, J.B., Orea, J.M., Montero, C., Urena, A.G., Navas, E., Slowing, K.,
Gomez-Serranillos, M.P., Carretero, E., De Martinis, D., 2005. Resveratrol
treatment controls microbial flora, prolongs shelf life, and preserves
nutritional quality of fruit. J. Agric. Food Chem. 53, 1526–1530.
Jitareerat, P., Paumchai, S., Kanlayanarat, S., Sangchote, S., 2007. Effect of chitosan
on ripening, enzymatic activity, and disease development in mango (Mangifera
indica) fruit. N. Z. J. Crop Hort. Sci. 35, 211–218.
163
Kondo, S., Kittikorn, M., Kanlayanarat, S., 2005. Preharvest antioxidant activities of
tropical fruit and the effect of low temperature storage on antioxidants and
jasmonates. Postharvest Biol. Technol. 36, 309–318.
López-Mora, L.I., Gutiérrez-Martínez, P., Bautista-Baños, S., Jiménez-García, L.F.,
Zavaleta-Mancera, H.A., 2013. Evaluation of antifungal activity of chitosan in
Alternaria alternata and in the quality of ‘Tommy atkins’ mango during storage.
Rev. Chapingo Ser. Hortic. 19, 315–331.
Mansour, M.M.F., 2000. Nitrogen containing compounds and adaptation of plants
to salinity stress. Biol. Plant. 43, 491–500.
Miller, G.L., 1959. Use of dinitrosalicylic acid reagent for the determination of
reducing sugar. Anal. Chem. 31, 426–429.
Miranda, M.V., Fernandez Lahor, H.M., Cascone, O., 1995. Horseradish peroxidase
extraction and purification by aqueous two-phase partition. Appl. Biochem.
Biotechnol. 53, 147–154.
Nawaz, K., Ashraf, M., 2009. Exogenous application of glycinebetaine modulates
activities of antioxidants in maize plants subjected to salt stress. J. Agron. Crop
Sci. 196, 28–37.
Ranganna, S., 1979. Manual of Analysis of Fruit and Vegetable Products, 2nd ed.
Tata McGraw-Hill Publishing Company Limited, New Delhi, 634 pp.
Robinson, S.P., Jones, G.P., 1986. Accumulation of glycine betaine in chloroplasts
provides osmotic adjustment during salt stress. Aust. J. Plant Physiol. 13,
659–668.
Romanazzi, G., Feliziani, E., Santini, M., Landi, L., 2013. Effectiveness of postharvest
treatment with chitosan and other resistance inducers in the control of storage
decay of strawberry. Postharvest Biol. Technol. 75, 24–27.
Sairam, R.K., Deshmukh, P.S., Shukla, D.S., 1997. Tolerance to drought and
temperature stress in relation to increased antioxidant enzyme activity in
wheat. J. Agron. Crop Sci. 178, 171–177.
Sivakumar, D., Jiang, Y., Yahia, E.M., 2011. Maintaining mango (Mangifera indica L.)
fruit quality during the export chain. Food Res. Int. 44, 1254–1263.
Sulaiman, S.F., Yusoff, N.A.M., Eldeen, I.M., Seow, E.M., Sajak, A.A.B., Supriatno,
O.K.L., 2011. Correlation between total phenolic and mineral contents with
antioxidant activity of eight Malaysian bananas (Musa sp.). J. Food Compos.
Anal. 24, 1–10.
Suseno, N., Savitri, E., Sapei, L., Padmawijaya, K.S., 2014. Improving shelf-life of
Cavendish banana using chitosan edible coating. Procedia Chem. 9, 113–120.
Yang, W.J., Rich, P.J., Axtell, J.D., Wood, K.V., Bonham, C.C., Ejeta, G., Mickelbart,
M.V., Rhodes, D., 2003. Genotypic variation for glycine betaine in sorghum.
Crop Sci. 43, 162–169.
Zhang, M., Zhang, H., Li, H., Lai, F., Li, X., Tang, Y., Min, T., Wu, H., 2016. Antioxidant
mechanism of betaine without free radicals scavenging ability. J. Agric. Food
Chem. 64, 7921–7930.
Zhang, Z., Yang, D., Yang, B., Gao, Z., Li, M., Jiang, Y., Hu, M., 2013. ˇ-Aminobutyric
acid induces resistance of mango fruit to postharvest anthracnose caused by
Colletotrichum gloeosporioides and enhances activity of fruit defense
mechanisms. Sci. Hortic. 160, 78–84.
Zhishen, J., Mengcheng, T., Jianming, W., 1999. The determination of flavonoid
contents in mulberry and their scavenging effects on superoxide radicals. Food
Chem. 64, 555–559.
Zhu, X., Wang, Q., Cao, J., Jiang, W., 2008. Effect of chitosan coating on postharvest
quality of mango (Mangifera indica L. cv. Tainong). J. Food Process Preserv. 32,
770–784.