Food Chemistry 196 (2016) 1040–1047

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Food Chemistry
journal homepage: www.elsevier.com/locate/foodchem

Pre-harvest application of polyamines enhances antioxidants and table

grape (Vitis vinifera L.) quality during postharvest period
S.H. Mirdehghan ⇑, S. Rahimi
Dep. Horticultural Sciences, College of Agriculture, Vali-e-Asr University of Rafsanjan, Kerman, Iran

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

Article history: Pre-harvest foliar spraying of grapevines with putrescine (Put) and spermidine (Spd) (0, 1, 2 mM) was
Received 11 July 2015 evaluated for determining the fruit quality at harvest and improving postharvest characteristics of table
Received in revised form 10 October 2015 grapes during cold storage. Fruit parameters in terms of firmness, fungal infection, weight loss, total phe-
Accepted 10 October 2015
nol concentration, antioxidant activity, skin color, total anthocyanin concentration, total soluble solids
Available online 22 October 2015
(TSS) and titratable acidity (TA) were evaluated after 0, 25 and 55 days of storage at 1.5 ± 1 °C and
90 ± 5% R.H. Softening, fungal infection and weight loss increased during cold storage but the rate of
Keywords:
changes significantly was delayed in Put- and Spd-treated fruits. Besides these, the application of Put
Anthocyanin
Fungal infection
and Spd maintained higher values of phenolics content, antioxidant activity and anthocyanins at the
Phenolics end of storage in compare to control. Furthermore, after 55 days of storage all treated fruits with Put
Spermidine and Spd showed lower changes in L⁄, TSS and TA and also higher value of chroma in berries.
Ó 2015 Elsevier Ltd. All rights reserved.

1. Introduction than control which leads to an increased shelf life of strawberry

Table grape (Vitis vinifera L.) is one of the most important fruit
that widely cultivated in the world and have been considered as
an exportable fruit from many countries. Although table grapes
have been classified as a non-climacteric fruit, they are highly per-
ishable due to high softening, weight loss and fungal decay, which
consequently leads to low storability. Different strategies have
been postulated to maintain firmness and control decay of table
grapes during storage and improve functional properties of fruit
such as pre and postharvest chitosan coating (Meng, Li, Liu, &
Tian, 2008) and exogenous abscisic acid application (Sandhu,
Gray, Lu, & Gu, 2011). Polyamines (PAs) application also showed
a significant role in extending the storage periods of several fruit
species with maintenance of fruit quality. Higher firmness and
lower weight loss was reported in many PAs-treated fruits such
as lemon (Valero, Martinez-Romero, & Riquelme, 1998), peach
and nectarine (Bregoli et al., 2002) and pomegranate
(Mirdehghan, Rahemi, Castillo et al., 2007) during storage.
Zokaee-Khosroshahi, Esna-Ashari, and Ershadi (2007) found that
PAs may influence fungal infection of strawberry fruit during stor-
age. They showed that Put-treated fruits had less fungal infection
⇑ Corresponding author.
E-mail address: mirdehghan@vru.ac.ir (S.H. Mirdehghan).
fruits from 7 to 10 to 12–14 days.
Postharvest treatments are not necessarily the best way of
maintaining fruit quality during postharvest period. Such treat-
ments are expensive, increase the risk of fruit damage through
extra handling and also encourage grower to pay less attention
to on-tree quality. Pre-harvest application considered as a good
alternative to cope with mentioned problem. However, the data
on pre-harvest application of polyamine on different fruit species
are scant. Khan, Singh, and Abbasi (2007) showed that pre-
storage application of Put would retard fruit softening in ‘Angelino’
plum during cold storage by suppressing ethylene biosynthesis. In
mango, Malik and Singh (2005) reported that pre-harvest applica-
tion of PAs improved fruit shelf life, increased ascorbic acid content
and retarded fruit skin color changes compared to control. To the
best of our knowledge, there is not any report in literature about
the role of pre-harvest application of PAs in table grape as a possi-
ble role in reducing mechanical damage of berries which leads to
lower decays. Besides these, damage caused by human handling
starts at harvest operation, which still occurs by hand for most
fruits. Thus, the objective of this work was to study the possible
role of pre-harvest foliar spraying of Put and Spd in maintenance
of table grape quality at harvest and during postharvest periods.
Two cultivars that have been included in this study were ‘Olhoghi’
characterized by a purple-red skin and ‘Rishbaba’ by a yellow skin
color. These cultivars are seeded, commercially valuable and have a
good storability.

http://dx.doi.org/10.1016/j.foodchem.2015.10.038
0308-8146/Ó 2015 Elsevier Ltd. All rights reserved.
 S.H. Mirdehghan, S. Rahimi / Food Chemistry 196 (2016) 1040–1047
2. Material and methods
The experiment was carried out on table grapes trees (V. vinifera
L. cvs. Olhoghi and Rishbaba) in a commercial orchard received
routine cultural practices. The trees (9-year-old) were spaced
2.5
2 m between and along the rows respectively in a head train-
ing system. twenty trees of each cultivar were selected for uniform
size and fruit load, and used for treatments by aqueous solutions
containing different concentrations of Put and Spd [0 (control), 1
and 2 mM] which were sprayed on the whole grapevines to run
off stage. Control trees only received an aqueous solution contain-
ing the same concentration of surfactant (Tween-20, 2 g L1). Each
tree was sprayed twice at 40 and 20 days before estimated harvest
time. Experimental layout was randomized block design and a sin-
gle tree represented as an experimental unit with four replications.
Fruits were considered during last stages of ripening and all
treated and control fruits harvested at commercial maturity basis
[TSS (15.2% and 17.25% for ‘Rishbaba’ and ‘Olhoghi’ respectively)
and titratable acidity (0.43% and 0.54% for ‘Rishbaba’ and ‘Olhoghi’
respectively]. Fruit samples then transported to the postharvest
laboratory, where different fruit grape clusters of yellow-green to
yellow in ‘Rishbaba’ and red color in ‘Olhoghi’, without damage
or pathological defect were selected and stored (grape clusters
weighing approximately 350–400 g in each experimental unit) at
1.5 ± 1 °C and 90 ± 5% R.H. Firmness, antioxidant activity, Phenolics
content, anthocyanins, berries color, total soluble solids (TSS) and
titratable acids (TA) were evaluated after 0, 25 and 55 days of stor-
age. Weight loss and microbial activity were measured after 25 and
55 days of cold storage.
2.1. Firmness
Berry firmness was recorded from 10 berries of each replicate
by using a digital pressure tester (model Lutron FG5020, Taiwan)
fitted with an 8 mm tip in diameter which was pressed gradually
on the surface of berries with nearly constant speed until the juice
exudation of individual berry and means were expressed as Kgf.
2.2. Microbial analysis
10 g of sample from each replicate was homogenized with
90 ml of sterile peptone water for 90 s using a stomacher blender
(Bag Mixer 400, InterScience, France). Serial dilutions of fruit
homogenates were prepared and 1 ml of final preparation was
poured in plate count agar under sterile conditions (Valverde
et al., 2005). Plates were incubated for 5 days at 25 °C and counts
of colony forming were considered and results were expressed as
Log CFU g1. All tests were run in duplicate.
2.3. Weight loss, color, TSS and TA
Weight of grape clusters for each treatment was recorded at 0,
25 and 55 days of storage and cumulative weight loss was calcu-
lated as the percentage loss of the initial weight.
Berry color (L⁄, a⁄ and b⁄ values) was determined with a Minolta
Chroma Meter CR-400 (Osaka, Japan). Color measurements were
performed using the Hunter Lab System. Color was expressed as
changes in chroma index [=(a⁄2 + b⁄2)0.5] and L⁄ during cold storage.
Color values were obtained for 10 berries per replicate. Three mea-
surements were taken from equatorial region of each berry. TSS
content in 10 berries of each clusters was assessed by a digital
refractometer (ATAGO, PAL-1 model Japan), and the results were
expressed as a percent. TA was determined by titration with
0.1 N NaOH, using 5 ml of diluted juice and expressed as g tartaric
acid 100 mL1 juice.
1041
2.4. Total phenolic compounds (TPC) and total antioxidant activity
(TAC)
5 g of frozen sample (in liquid nitrogen and kept at 80 °C) in
the whole berry of grape fruits was homogenized in 10 ml of
50 mM phosphate buffer pH = 7.8 and centrifuged at 13,000 rpm
at 4 °C for 20 min. The supernatant was used for TAC and TPC anal-
ysis. Folin–Ciocalteu method was used to determine TPC according
to Serrano, Guillen, Martinez-Romero, Castillo, and Valero (2005),
with some modification. 100 ll of supernatant was mixed with
400 ll phosphate buffer and 2.5 ml of folin. After 1 min, 2 ml of
sodium carbonate solution (7.5%) was added to the mixture and
allowed to stand for 5 min at 50 °C. Absorbance was read at
760 nm with a spectrophotometer (Helios Zeta UVVIS, Thermo
Fisher Scientific, UK). Gallic acid was used as standard to produce
the calibration curve, and results were expressed as mg gallic acid
equivalent 100 g1 of fresh weight.
TAC was carried out as described by Serrano et al. (2005), with
some modification. Briefly, TAC was determined using the enzy-
matic system composed of the chromophore 2,20 -azinobis-(3-ethyl
benzothiazoline-6-sulfonic acid) diammonium salt (ABTS), the
horse radish peroxidase enzyme, and its oxidant substrate (hydro-
gen peroxide), in which ABTS+ radicals are generated and moni-
tored at 414 nm. The decrease in absorbance after adding the
supernatant was proportional to TAC of the sample. A calibration
curve was performed with L-ascorbic acid (0–20 nmol) from Sigma
(Poole, Dorset, United Kingdom), and results are expressed as mg
of L-ascorbic acid equivalent 100 g1 fresh weight.
2.5. Total anthocyanins
Differential pH method was used for determination of total
anthocyanin contents (Wrolstad, 1976). 5 g of homogenized fruit
were centrifuged at 10,000 rpm and hydrochloric acid–potassium
chloride (pH = 1) and acetate (pH = 4.5) buffers were used for dilu-
tion of supernatants. Absorbance was measured in a UV–Vis spec-
trophotometer at 520 nm and at 700 nm in buffers at pH 1 and 4.5
respectively. The absorbance was calculated by A = ((A520  A700)-
pH1  (A520  A700)pH4.5) with a molar extinction coefficient of
malvidin-3-glucoside of 28,000 L mol1 cm1. The results were
expressed as mg of malvidin-3-glucoside equivalent per kilogram
fresh weight.
2.6. Statistical analysis
Each value is the average of four replications. The experimental
data were subjected to analysis of variance (ANOVA) using MSTATC
statistical software and the means were compared by Tukey test.
Sources of variation were time of storage and treatments. Correla-
tion analysis was performed between some parameter taking into
account all sampling data (0, 25 and 55 days of cold storage).
3. Results and discussions
3.1. Firmness
Significant higher firmness of berries was observed in Put- and
Spd-treated table grape in compare to control, which was detect-
able immediately after harvest in each cultivar (Fig. 1). The highest
levels were observed in Spd 1 mM (0.58 Kgf) and Put 2 mM
(0.7 Kgf) in ‘Rishbaba’ and ‘Olhoghi’ cultivars respectively. Fruit
firmness declined during storage, but was always higher in PAs-
treated fruits than control. The rate of fruit softening could be
divided into two different stages, which is slow until 25 days of
cold storage followed by accelerated decline in fruit firmness up
1042
Cont rol
P ut (1 mM)
0.8
P ut (2 mM)
Spd (1 mM)
Spd (2 mM)
c-f
0.6
Firmness (Kg f)
g-i
h-j
k-n
l-n
0.4
0.2
0.0
0 25
Values with similar letters are not significantly different (p < 0. 05).
Fig. 1. Effect of pre-harvest foliar spray of Put and Spd on firmness of table grape cultivars of Rishbaba and Olhoghi stored at 1.5 ± 1 °C. Data are means ± S.E.
to the end of cold storage. At the end of experiment (55 days of
storage) the loss of firmness was indicated in all fruits especially
in control while the treated fruits showed firmer berries, especially
Spd-treated berries was the most effective in maintaining the firm-
ness than others in both cultivars.
Accelerated softening of table grape were limited the storage
potential and reduce the quality of the table grape. The changes
were significantly delayed in those berries treated with Put or
Spd during cold storage. Some of these effects even could be
detectable at harvest (0 days of storage), indicating that both Put
and Spd pre-harvest spraying was able to maintain the firmness
of berries compared to control. Similarly, Khan et al. on plum
(2007) and Malik and Singh on mango (2005) observed increased
firmness of fruits at harvest by pre-harvest treatments of different
PAs. Also delay of softening process was reported during cold stor-
age in several fruits including plums (Serrano, Martinez-Romero,
Guillen, & Valero, 2003), lemon and mandarin (Valero et al.,
1998). Softening contributes to quality loss in reducing the shelf
life, but PAs treatment resulted in maintenance of flesh firmness
during cold storage. Therefore, Put- and Spd-treated grape have
higher firmness at harvest leading to much lower mechanical
injury during harvest and handling process and providing better
transportability.
Deng, Wu, and Li (2005) showed a dramatic decrease in hemi-
celluloses (38–63%) and moderate decreases in cellulose (9–16%)
and total pectin (7–15%) in grapes ‘Kyoho’ stored in different atmo-
spheric condition at air and 0 °C. This indicates that the softening
of grapes resulted from an increase in depolymerization and degra-
dation of cell wall polysaccharides. Khan et al., 2007 found an
inverse relationship between cell wall degrading enzymes activity
and Pre- and post-harvest Put application leading to a higher firm-
ness and lower activity of exo-PG, endo-PG, PE and EGase in skin
and pulp tissues of treated plum during cold storage. Also it has
been reported that the role of PAs in delaying softening of peach
could be attributed to their effects on stabilization of cell wall
(Bonghi, Ferrarese, Ruperti, Tonutti, & Ramina, 1998).
Table grape classified as a susceptible fruit to mechanical dam-
ages. In developing countries, there are not much suitable facilities
at harvest and handling period to avoid mechanical damage and
eventually leads to higher commercial losses. Therefore the role
S.H. Mirdehghan, S. Rahimi / Food Chemistry 196 (2016) 1040–1047
Rishbaba Olhoghi
a ab
cd a-c
bc
c-e c-e
c-f
e-g d-g
f-h
g-i
h-j h-k
i-k k-n i-l
j-m
l-n
mn n
o
op
op p
55 0 25 55
Storage time (Days)
of preharvest PAs treatment is very valuable and effective on main-
taining fruit firmness subsequently lead to reduced losses by
mechanical damage.
3.2. Weight loss and microbial activity
As shown in Fig. 2A and B cumulative weight loss of fruits
increased during storage in both cultivars although it was signifi-
cantly higher in control than in PAs-treated bunches. The effects
were more obvious after 55 days of storage where the Spd concen-
trations decreased more than 50% of weight loss than in control.
Pre-harvest foliar spray of grape fruits with Spd and Put signif-
icantly reduced the microbial activity of berries during storage per-
iod at 1.5 ± 1 °C in both cultivars (Fig. 2C and D). After 55 days of
storage, Spd 1 mM with an average of 3.22 Log CFU g1 in ‘Rish-
baba’ (Fig. 2C) and Spd 2 mM (3.27 Log CFU g1) in ‘Olhoghi’
(Fig. 2D) has known as the most effective treatments on controlling
microbial activity of stored berries.
The berries of control treatment experienced an acceleration of
weight loss during cold storage, which can be attributed to
increased level of microbial activity, associated with tissue senes-
cence over long storage times, which is slowed down following
Put and Spd application. In this sense, we found a significant pos-
itive correlation between fungal infection and weight loss of table
grapes (r = 0.76⁄⁄). Previous studies have reported the effectiveness
of PAs treatment in reducing weight loss of pomegranate
(Mirdehghan, Rahemi, Castillo et al., 2007). It was suggested that
PAs conjugate to the membrane phospholipids led to cell mem-
brane integrity, which resulted the lower weight loss that was sug-
gested in some horticultural commodities (Artes-Hernandez &
Aguayo, 2004; Woods, 1990).
In table grapes, mechanical injury of berries leads to develop-
ment of fungal pathogens during handling and storage. It is well
known that one of the major postharvest problems of table grape
is its susceptibility to gray mold which caused by Botrytis cinerea
eventually limited the shelf life during storage and retail marketing
(Romanazzi, Lichter, Gabler, & Smilanick, 2012). Fungal decay
would affect appearance characteristics, firmness and weight loss
of fruits during cold storage.
 S.H. Mirdehghan, S. Rahimi / Food Chemistry 196 (2016) 1040–1047 1043

Cont rol
a
8 P ut (1 mM) (A) (B)
P ut (2 mM) Rishbaba Olhoghi
Spd (1 mM ) a
Spd (2 mM)
b
c
6

Weight loss (%)

d
e
f f e
4
g g
g

h
ij ij hi
2 k-l jk
l kl

0
5 (C) (D)
Rishbaba Olhoghi
Microbial activity (Log CFUg )
-1

a ab
4 ab
a-d a-c
b-d b-d
cd d cd
d de
3 ef
f ef
f f
f f
g
2

0
25 55 25 55

Storage time (Days)

Values with similar letters are not significantly different (p < 0. 05).
Fig. 2. Effect of pre-harvest foliar spray of Put and Spd on weight loss (A and B) and microbial activity (C and D) of table grapes stored at 1.5 ± 1 °C. Data are means ± S.E.

Different methods were demonstrated to control table grapes activity during cold storage (Fig. 3B), while in ‘Rishbaba’, the levels
postharvest decay, such as chitosan film (Meng et al., 2008; of antioxidant activity was higher in Put 2 mM and Spd 1 mM at
Romanazzi et al., 2012), coating with Aloe vera gel (Valverde 25 days and Spd 2 mM after 55 days of storage (Fig. 3A).
et al., 2005), modified atmosphere packaging (Artes-Hernandez The changes in total phenolics content of berries was nearly
and Aguayo, 2004). Our results indicates an increase of fungal constant during storage, the level being slightly decreased as the
infection of berries during cold storage but was significantly lower storage time progressed. However during cold storage, phenolics
in Put- and Spd-treated fruits. The results are in accordance with content was affected by different concentration of Spd and Put
that study on strawberry cv. Selva, in which Put-treated fruits treatments, in which the lowest value was observed in control
had less fungal infection symptoms than untreated ones and main- and the highest content was in fruit treated with Spd 1 mM in
tained the fruits with a very good appearance. ‘Rishbaba’ (Fig. 3C) and Spd 2 mM in ‘Olhoghi’ (Fig. 3D). The level
Few reports are available on antimicrobial activity of PAs on was markedly higher in Put- or Spd-treated berries than those in
horticultural crops and its interaction between plant and patho- control after 55 days of storage.
gens. However, it has been shown that the increased levels of Higher levels of phenolic compounds and antioxidant activity
PAs accompanied with lower infection of barley leaves with brown were observed in Put-and Spd-treated berries of table grape than
rust (Puccinia hordei) and powdery mildew fungi (Blumeria grami- control. Mirdehghan, Rahemi, Serrano et al. (2007) reported the
nis F. sp. Hordei) (Greenland & Lewis, 1984), therefore it was con- same increase of phenolics and antioxidants in pomegranate fruits
cluded that PAs may have a role in plant defense against treated with Spd or Put. Phenolic acids (cinnamic and benzoic,
pathogens but the involved molecular mechanisms are still esterified or not with tartaric acid) are mainly present in white
unclear. grapes. These compounds are highly oxidative, producing brown
compounds that also show antioxidant activity (Jimenez et al.,
3.3. Antioxidant activity and phenolics 2002).
The antioxidant properties of PAs has been shown in many
Antioxidant activity of berries significantly influenced by Put- studies implying that PAs exhibited their most significant antioxi-
and Spd-treated grape, in which the value were higher in treated dant properties when they form conjugates with phenolic acids
berries than those in control throughout postharvest periods (Bors, Langebarteles, Michel, & Sanderman, 1989). Thus, higher
(Fig. 3A and B). The effectiveness of treatments was depends on levels of these compounds in grape fruits treated with PAs could
the type of PAs and cultivars. In ‘Olhoghi’, Put 2 mM was more be attributed to antioxidant properties of PAs. Besides these, it
effective than other treatments in maintenance of antioxidant was observed a negative significant correlation between phenolic
1044 S.H. Mirdehghan, S. Rahimi / Food Chemistry 196 (2016) 1040–1047

Control (A) (B)

P ut (1 mM) Rishbaba Olhoghi
(mg ascorbic acid equiv 100g-1 FW)
60 P ut (2 mM) a
Spd (1 mM )
a a
Spd (2 mM)
Antioxidant activity

ab
b-d
b-e
b-f bc b-d b-e
b-f
40 b-f b-g
e-j c-i c-h e-j
e-k d-i i-m
h-l f-l g-l
h-m
j-m i-m i-m
k-m lm
m
20

0
(C) Rishbaba Olhoghi a (D)
a-d a-d ab a-c
60 a-d a-e a-d
(mg equiv gallic acid 100g-1)

a-c a-f
b-h b-h b-g
b-h
b-g b-g b-g b-g b-h b-g a-f
e-h d-h
Phenolic compounds

d-h c-h
gh g-h f-h e-h

h
40

20

0
0 25 55 0 25 55

Storage time (Days)

Values with similar letters are not significantly different (p < 0. 05).
Fig. 3. Effect of pre-harvest foliar spray of Put and Spd on antioxidant activity (A and B) and phenolics (C and D) of table grape stored at 1.5 ± 1 °C. Data are means ± S.E.

compounds and antimicrobial activity of treated and untreated be concluded that higher value of color intensity in PAs-treated
table grape (r = 0.51⁄⁄⁄). berries was due to their more anthocyanins concentrations.

3.4. Lightness and chroma 3.5. Total anthocyanins

‘Olhoghi’ table grape is characterized by a purple–red skin color As noted before, ‘Olhoghi’ is a purple–red skin color cultivar and
and ‘Rishbaba’ by a yellow skin color at the optimum stage of har- showed a mean of 49.19 ± 3.3 mg kg1 of anthocyanins at harvest,
vest. Color coordinate in ‘Olhoghi’ were a⁄ = 6.48 ± 0.69, while it was not detectable in ‘Rishbaba’ cultivar and therefore
b⁄ = 1.46 ± 0.14, L⁄ = 28.16 ± 0.39 and in ‘Rishbaba’ they were only the data of former cultivar was considered here. During cold
a⁄ = 5.72 ± 0.36, b⁄ = 15.43 ± 0.93, L⁄ = 46.05 ± 0.5 in control treat- storage, levels of these pigments increased by 25 days of storage
ment at harvest. However, pre-harvest Spd- and Put-treated grape and then declined over the end of storage time, but it was signifi-
berries exhibited higher L⁄ during 25 and 55 days of storage com- cantly higher in all Put- and Spd-treated berries at the end of
pared to untreated fruit especially at the end of cold storage experiment (Fig. 5). On the contrary, the level of total anthocyanins
(Fig. 4A and B). Spd 1 and 2 mM showed highest value of L⁄ in both was higher in control than treated berries after 25 days of storage
cultivars at the end of experiments. During cold storage, control and may be results from the delay-effect of PAs treatment on
berries showed the greater variations of chroma and all these vari- ripening process.
ation were lower in Put- and Spd-treated grape. Similarly, the high- The purple skin color of table grape was related to the presence
est chroma related to Spd 1, 2 mM in both cultivars compared to of anthocyanin compounds, from which the anthocyanin malvidin-
control fruit (Fig. 4C and D). 3-glucoside has been found as major component (Somers, 1966).
Little changes were observed in color of mature yellow–white Although, total anthocyanins were reduced in control and treated
and red–purple grape berries during storage, but during long- fruits during cold storage, but pre-harvest foliar spraying of Put
term storage, berry browning may occur (Hardenburg, Watada, & or Spd delayed total anthocyanins concentration after 25 days of
Wang, 1990). Romero, Teresa Sanchez-Ballesta, Maldonado, Isabel storage and decreased the loss of these compounds at the end of
Escribano, and Merodio (2008) noted that decrease in anthocyanin experiment. PAs have been described as anti-senescence agents
levels in table grape cv. Cardinal match with a decrease in L⁄ and and a great number of researches have been focused on the role
chroma values. Similar trends and rates of changes in antho- of exogenous PAs on fruit ripening (Valero, Martinez-Romero, &
cyanins, lightness and chroma were also found for our samples. Serrano, 2002). Also it has been reported that the ripening process
The higher the anthocyanins concentration in grape berries, the and senescence of table grapes is correlated with the anthocyanin
higher value of lightness and chroma is likely to be. Thus, it can concentration and profile (Cantos, Espin, & Tomas-Barberan, 2002).
 S.H. Mirdehghan, S. Rahimi / Food Chemistry 196 (2016) 1040–1047 1045

Control
P ut (1 mM) (A) Rishbaba Olhoghi (B)
P ut (2 mM)
60 Spd (1 mM )
Spd (2 mM)

ab a ab ab a-d ab a-d
a-c a-d
b-d a-d de c-e
40 f ef
L*

gh g-i g
g-i g-i g g-i g-j g-j
g-i ij h-j
jk
k k
20

0
60 (C) Rishbaba Olhoghi
(D)

ab
50 a

bc
40 bc
b-d
Chroma

cd
de
30 d-f
d-g
e-h
e-i
hi e-i f-i e-i
20 f-i g-i hi hi hi e-i
hi hi i i hi hi hi
hi
i

10

0
0 25 55 0 25 55

Storage time (Days)

Values with similar letters in each column are not significantly different (p < 0. 05).
Fig. 4. Effect of pre-harvest foliar spray of Put and Spd on L⁄ (A and B) and chroma (C and D) of table grapes stored at 1.5±. Data are means ± S.E.

Cont rol a
80 Put (1 mM)
P ut (2 mM)
Spd (1 mM)
ab
Spd (2 mM)
a-c
b-d b-d
60 b-d
b-e b-d
b-e b-e
c-e
Anthocyanin

b-e
(mg kg-1)

de c-e

40 e

20

0
0 25 55

Storage time (Days)

Values with similar letters in each column are not significantly different (p < 0. 05).

Fig. 5. Effect of pre-harvest foliar spray of Put and Spd on anthocyanin in table grapes ‘Olhoghi’ stored at 1.5 ± 1 °C for 25 and 55 days. Data are means ± S.E.
1046 S.H. Mirdehghan, S. Rahimi / Food Chemistry 196 (2016) 1040–1047

Khan & Singh, 2010 concluded that pre-harvest application of Put
2 mM delayed the ripening process in three cultivars of Japanese
plum with reduced respiration rate, ethylene production and fruit
softening. Thus the higher concentration of anthocyanins in PAs-
treated berries at the end of storage could be related to its effect
on ripening and senescence delay of treated table grape.
observed, as generally reported for grape fruits like ‘Perlit’ which
accounted for the higher TSS/TA ratio over storage. Delay in ripen-
ing is one of the effects of PAs properties and reported in many
fruits such as nectarine, Japanese plum (Khan & Singh, 2010) and
mango (Malik & Singh, 2005).

3.6. Total soluble solids (TSS) and titratable acidity (TA) 4. Conclusion

TSS and TA were affected by pre-harvest foliar application of Put
and Spd solution. A lower TSS% content and higher TA% was
observed in Put- and Spd-treated berries of grape than control at
harvest except some concentration (Fig. 6), indicating a marked
delay in process of maturation and ripening. During cold storage,
similar pattern of changes in TSS% and TA% content were observed,
in which higher value of TA% and lower value of TSS% were
detected in Put- and Spd-treated berries resulting different ripen-
ing index, which were significantly higher in control than treated
berries.
The main taste change during storage is a slight sweetening and
a loss of acidity which can cause the fruits to taste insipid. The total
soluble solids of grapes are largely made up of glucose and fructose
and the predominant acids are tartaric and malic acid (Kanellis &
Roubelakis-Angelakis, 1993). Pre-harvest application of Put and
Spd delayed the process of ripening in table grapes. During cold
storage, an increase in TSS and decrease in TA of grape juice were
Pre-harvest foliar application of Put and Spd on grapevines
maintained higher firmness at harvest and postharvest periods
and also improved the fruit quality in terms of antioxidant activity,
phenolics, anthocyanin and also controlling weight loss during cold
storage. Taking into account the overall results obtained, the most
effective treatment was Spd that showed the highest maintenance
of firmness and also the lowest antimicrobial activity of berries at
the end of cold storage. Since the fungal decay and mechanical
damage considered as the main causes of low storability in table
grape, using these naturally occurring substances can be a useful
tool for controlling fruit losses. In the USA, there is a patent
(Law, Davies & Mutschler, 1988) for the use of PAs as method of
extending shelf life and enhancing keeping quality of fruits. There-
fore, the beneficial effect of PAs treatments are commercially sig-
nificant and alleviate the risk of mechanical damage at harvest
and during handling and eventually could be a good alternative
in table grapes during marketing even at retail level.

25
Control
(A) Rishbaba Olhoghi (B)
Put (1 mM)
Put (2 mM)
Spd (1 mM )
20 Spd (2 mM) b a
cd c
de
fg fg ef g g g g
h-j h h-k hi hi
i-k h-j k jk jk
l m lm mn m lm
15
no o
TSS

10

0
(C) Rishbaba Olhoghi (D)
a
0.8 ab a-c
b-e c-f b-f b-c
d-g
g-j e-h f-h g-j
g-i f-h g-j
g-j
0.6 h-k
i-l j-m
m-o k-n l-o
TA (%)

k-n
n-p
op p p
0.4
q
q
q
0.2

0.0
0 25 55 0 25 55

Storage time (Days)

Values with similar letters in each column are not significantly different (p < 0. 05).
Fig. 6. Effect of pre-harvest foliar spray of Put and Spd on TSS (A and B) and TA (C and D) of table grapes stored at 1.5±. Data are means ± S.E.
 S.H. Mirdehghan, S. Rahimi / Food Chemistry 196 (2016) 1040–1047
Acknowledgement
The research was supported by Vali-e-Asr University of
Rafsanjan.
References
Artes-Hernandez, F., & Aguayo, E. (2004). Alternative atmosphere treatments for
keeping quality of autumn seedless table grapes during long term cold storage.
Postharvest Biology and Technology, 31, 59–67.
Bonghi, C., Ferrarese, L., Ruperti, B., Tonutti, P., & Ramina, A. (1998). Endo-1,4-
glucanases are involved in peach fruit growth and ripening, and regulated by
ethylene. Plant Physiology, 102, 346–352.
Bors, W., Langebarteles, C., Michel, C., & Sanderman, H. (1989). Polyamines as
radical scavengers and protectants against ozone damage. Phytochemistry, 28,
1589–1595.
Bregoli, A. M., Scaramagli, S., Costa, G., Sabatini, E., Ziosi, V., Biondi, S., & Torrigiani, P.
(2002). Peach (Prunus persica L.) fruit ripening: Aminoethoxyvinylglycine (AVG)
and exogenous polyamines affect ethylene emission and flesh fimness. Plant
Physiology, 114, 472–481.
Cantos, E., Espin, J. C., & Tomas-Barberan, F. A. (2002). Varietal differences among
the polyphenol profiles of seven table grape cultivars studied by LC-DAD-MS-
MS. Journal of Agricultural and Food Chemistry, 50, 5691–5696.
Deng, Y., Wu, Y., & Li, Y. (2005). Changes in firmness, cell wall composition and cell
wall hydrolases of grapes stored in high oxygen atmospheres. Food Research
International, 38, 769–776.
Greenland, A. J., & Lewis, D. H. (1984). Amines in barley leaves infected with brown
rust and their possible relevance to formation of ‘green islands. New Phytologist,
96, 283–291.
Hardenburg, R. E, Watada, A. E., Wang, C. Y. (1990). The commercial storage of fruits,
vegetables, and florist and nursery stocks, U.S. Department of Agriculture,
Agriculture Handbook No 66.
Jimenez, A., Creissen, G., Kular, B., Firmin, J., Robinson, S., Verhoeyen, M., &
Mullineaux, P. (2002). Changes in oxidative processes and components of the
antioxidant system during tomato fruit ripening. Planta, 214, 751–758.
Kanellis, A. K., & Roubelakis-Angelakis, K. A. (1993). Grape. In G. B. Seymour, J. E.
Taylor, & G. A. Tucker (Eds.), Biochemistry of fruit ripening (pp. 189–234).
London, UK: Chapman and Hall.
Khan, A. S., & Singh, Z. (2010). Pre-harvest application of putrescine influences
japanese plum fruit ripening and quality. Food Science and Technology
International, 16, 53–64.
Khan, A. S., Singh, Z., & Abbasi, N. A. (2007). Pre-storage putrescine application
suppresses ethylene biosynthesis and retards fruit softening during low
temperature storage in ‘Angelino’ plum. Postharvest Biology and Technology,
46, 36–46.
Law, D. M., Davies, P. J., Mutschler, M. A. (4957757). Method of extending shelf life
and enhancing keeping quality of fruits. US patent, 1988.
Malik, A. U., & Singh, Z. (2005). Pre storage application of polyamines improves shelf
life and fruit quality of mango. Journal of Horticultural and Science and
Biotechnology, 80, 363–369.
1047
Meng, X., Li, B., Liu, J., & Tian, S. (2008). Physiological responses and quality
attributes of table grape fruit to chitosan preharvest spray and postharvest
coating during storage. Food Chemistry, 106, 501–508.
Mirdehghan, S. H., Rahemi, M., Castillo, S., Martinez-Romero, D., Serrano, M., &
Valero, D. (2007). Pre-storage application of polyamines by pressure or
immersion improves shelf life of pomegranate stored at chilling temperature
by increasing endogenous polyamine levels. Postharvest Biology and Technology,
44, 26–33.
Mirdehghan, S. H., Rahemi, M., Serrano, M., Guillen, F., Martinez-Romero, D., &
Valero, D. (2007). The application of polyamines by pressure or immersion as a
tool to maintain functional properties in stored pomegranate arils. Journal of
Agricultural and Food Chemistry, 55, 755–760.
Romanazzi, G., Lichter, A., Gabler, F. M., & Smilanick, J. L. (2012). Recent advances on
the use of natural and safe alternatives to conventional methods to control
postharvest gray mold of table grapes. Postharvest Biology and Technology, 63,
141–147.
Romero, I., Teresa Sanchez-Ballesta, M., Maldonado, R., Isabel Escribano, M., &
Merodio, C. (2008). Anthocyanin, antioxidant activity and stress-induced gene
expression in high CO2-treated table grapes stored at low temperature. Journal
of Plant Physiology, 165, 522–530.
Sandhu, A. K., Gray, D. J., Lu, J., & Gu, L. (2011). Effects of exogenous abscisic acid on
antioxidant capacities, anthocyanins, and flavonol contents of muscadine grape
(Vitis rotundifolia) skins. Food Chemistry, 126, 982–988.
Serrano, M., Guillen, F., Martinez-Romero, D., Castillo, S., & Valero, D. (2005).
Chemical constituents and antioxidant activity of sweet cherry at different
ripening stages. Journal of Agricultural and Food Chemistry, 53, 2741–2745.
Serrano, M., Martinez-Romero, D., Guillen, F., & Valero, D. (2003). Effects of
exogenous putrescine on improving shelf life of four plum cultivars. Postharvest
Biology and Technology, 30, 259–271.
Somers, T. C. (1966). Grape phenolics: the anthocyanins of Vitis vinifera, var. Shiraz.
Journal of the Science of Food and Agriculture, 17, 215–219.
Valero, D., Martinez-Romero, D., & Riquelme, F. (1998). Influence of postharvest
treatment with putrescine and calcium on endogenous polyamines, firmness,
and abscisic acid in lemon (Citrus lemon L. Burm cv. Verna). Journal of
Agricultural and Food Chemistry, 46, 2102–2109.
Valero, D., Martinez-Romero, D., & Serrano, M. (2002). The role of polyamines in the
improvement of the shelf life of fruit. Trends in Food Science & Technology, 13,
228–234.
Valverde, J. M., Valero, D., Martinez-Romero, D., Guillen, F., Castillo, S., & Serrano, M.
(2005). Novel edible coating based on Aloe vera gel to maintain table grape
quality and safety. Journal of Agricultural and Food Chemistry, 53, 7807–7813.
Woods, J. L. (1990). Moisture loss from fruits and vegetables. Postharvest News
Information, 1, 195–199.
Wrolstad, R. E. (1976). Color and pigment analysis in fruit products. Agricultural
Experiment Station, Oregon. State University, Station Bulletin, Corvallis, 624,
1–17.
Zokaee-Khosroshahi, M. R., Esna-Ashari, M., & Ershadi, A. (2007). Effect of
exogenous putrescine on post-harvest life of strawberry (Fragaria ananassa
Duch.) fruit, cultivar Selva. Scientia Horticultuare, 114, 27–32.