Research Article

Received: 20 January 2009 Revised: 27 May 2009 Accepted: 27 May 2009 Published online in Wiley Interscience: 9 July 2009

(www.interscience.wiley.com) DOI 10.1002/jsfa.3691

Effect of methyl jasmonate on quality

and antioxidant activity of postharvest loquat
fruit
Shifeng Cao,a,b Yonghua Zheng,a∗ Zhenfeng Yang,a Kaituo Wanga
and Huaijing Ruia

Abstract
BACKGOUND: Loquat fruit is rich in natural antioxidants and has shown a remarkably high antioxidant activity. To search for an
effective method for maintaining or even improving antioxidant activity during postharvest storage, we investigated the effect
of 10 µmol L−1 methyl jasmonate (MeJA) treatment on levels of major individual sugars and organic acids, total phenolics, total
carotenoids, total flavonoids and antioxidant activity in loquat fruit during storage at 1 ◦ C for 35 days.

RESULTS: The MeJA-treated fruit exhibited significantly lower levels of respiration rate, ethylene production, phenylalanine
ammonia-lyase and polyphenol oxidase activities, and higher levels of sugars, organic acids, total phenolics and total
flavonoids than control fruit. Meanwhile, the treatment also maintained significantly higher antioxidant activity as measured
by the scavenging capacity against 1,1-diphenyl-2-picrylhydrazyl (DPPH), superoxide and hydroxyl radicals, and by the reducing
power test compared to the control. There was a significant positive linear relationship between total phenolic content and
DPPH radical scavenging activity.

CONCLUSION: MeJA treatment can improve the quality and functional properties of harvested loquat fruit by maintaining a
higher level of antioxidants and enhancing antioxidant activity.


c 2009 Society of Chemical Industry

Keywords: loquat fruit; methyl jasmonate; quality; phenolics; flavonoids; antioxidant activity

INTRODUCTION
Free sugars and organic acids are natural components of many
fruits that play important roles in maintaining fruit quality and
determining nutritional value.1 The nature and the concentration
of these constituents in fruits have been of interest because of
their important influence on the organoleptic properties.
There is convincing epidemiological evidence showing that
a high consumption of fruits and vegetables can provide
protection against numerous chronic diseases, including cancer,
cardiovascular, cerebrovascular, ocular and neurological diseases.
This biological effect may be due largely to the presence
of antioxidant constituents, including vitamin C, vitamin E,
carotenoids, flavonoids and phenolic acids, as well as other
unidentified compounds.2,3 Therefore, these constituents have
received increasing attention for their antioxidant activity and the
level of antioxidant content has become an important parameter
with respect to fruit and vegetable quality.4
Interest in the role of free sugars and organic acids in fruit
quality and antioxidants in human health has promoted research
in the field of horticulture and food science to evaluate fruit
sugars, organic acids and antioxidants and to determine how their
content and activity can be maintained or even improved during
postharvest storage.
Loquat (Eriobotrya japonica Lindl.) is an evergreen tree native to
2064
yellow or white in color, with a soft and juicy flesh. The fruit is highly
favored by consumers worldwide for their mild, sub-acid and sweet
taste. Meanwhile, loquat fruit is also a good source of natural
antioxidants such as polyphenols, carotenoids and flavonoids,5
and has shown a remarkably high scavenging activity against
chemically generated radicals, thus making it effective in inhibiting
oxidation of human low-density lipoproteins.6 However, no studies
have been conducted to evaluate the effects of postharvest
conditions on the changes of phenolics and antioxidant activity
of loquat fruit, except for a report that loquat fruit stored at low
temperature had higher levels of total phenolics and 1,1-diphenyl-
2-picrylhydrazyl (DPPH) radical scavenging activity than the fruit
held at higher temperature.7
Methyl jasmonate (MeJA) is a naturally occurring plant growth
regulator that plays an important role in promoting biosynthesis of
secondary metabolites, inducing the expression of a set of defense
∗ Correspondence to: Yonghua Zheng, College of Food Science and Technology,
Nanjing Agricultural University, Weigang 1, Nanjing 210095, PR China.
E-mail: zhengyh@njau.edu.cn
a College of Food Science and Technology, Nanjing Agricultural University,
Weigang 1, Nanjing 210095, PR China

b Nanjing Research Institute for Agricultural Mechanization, Ministry of

China. The ripe fruit of loquat is spherical or oval in shape, orange- Agriculture, Liuying 100, Nanjing 210014, PR China

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c 2009 Society of Chemical Industry
Methyl jasmonate maintains quality and enhances antioxidant activity in loquat fruit
genes, and activating resistance of host against pathogens.8 In
recent research, it has been reported that a postharvest MeJA
treatment maintained higher levels of sugars and organic acids in
fresh-cut kiwifruit and radishes and enhanced antioxidant capacity
in blackberries, raspberries and strawberries.9 – 12 Thus MeJA has
a potential application in postharvest treatments for enhancing
antioxidant activity and maintaining a high-quality product. In a
previous study, we found that MeJA treatment at 10 µmol L−1 was
effective in reducing decay and maintaining quality of cold-stored
loquat fruit.13 However, there were no published data on the effect
of postharvest MeJA treatment on antioxidants and antioxidant
activity in loquat fruit. The objective of this work was to investigate
the effect of a pre-storage 10 µmol L−1 MeJA treatment on levels
of sugars, organic acids, total phenolics, total carotenoids and
flavonoids, antioxidant activity, and activities of phenylalanine
ammonia lyase (PAL) and polyphenol oxidase (PPO) in loquat fruit
during storage at 1 ◦ C.
MATERIALS AND METHODS
Plant material and treatments
Loquat fruit (Eriobotrya japonica Lindl. cv. Fuyang) were hand-
harvested at ripe stage from an orchard in Jiangsu, PR China, and
transported within 2 h to our laboratory. Fruit were selected for
uniform size and color and the absence of visual defects, then
randomly divided into two lots. The first lot of fruit was treated
with10 µmol L−1 MeJA (Aldrich Chemical Co., Milwaukee, WI, USA)
in sealed chambers at 20 ◦ C for 24 h, whereas the second lot of fruit
was subjected to the same conditions without exposure to MeJA
(control). The MeJA treatment was performed by placing a filter
paper soaked with MeJA solution into the container, and allowing
the MeJA to go into the container headspace. Fruit were exposed
to 10 µmol L−1 MeJA at 20 ◦ C for 24 h. For control fruit, a water-
soaked filter paper was used. Following treatment, the chambers
were opened, and both lots of fruit were stored (1 ◦ C, 95% relative
humidity) for up to 35 days. There were three replicates of 120
fruit each per treatment, and the experiment was conducted
twice. Twenty fruit samples were taken before MeJA treatment
(time 0) and at 7-day intervals during storage for measurements
of respiration rate, ethylene production, sugars, organic acids,
total phenolics, total flavonoids and total carotenoids, antioxidant
capacity and activities of PAL and PPO.
Measurement of respiration rate and ethylene production
Ten fruit for each of three replicates at each time point were
enclosed in 1 L glass jars at 1 ◦ C. 10 mL of headspace gas was
taken from each jar at the end of 2 h. CO2 was measured with an
infrared gas analyzer (GXH-305, Beijing, China). Respiration rate
was expressed as mL CO2 kg−1 fresh weight (FW) h−1 . Ethylene
was analyzed with a gas chromatograph with an alumina column
and a flame ionization detector. Results were expressed as µL
ethylene kg−1 FW h−1 .
Analysis of sugars and organic acids
Sugars and organic acids were extracted from loquat fruit using
the method as previously described,5 with some modifications.
5 g of flesh tissue was homogenized in 20 mL of 950 mL L−1 cold
ethanol solution for 1 min and shaken for 10 min. The homogenate
was filtered and the residue extracted twice with 800 mL L−1 cold
ethanol. The combined extracts were evaporated under vacuum
at 35 ◦ C until the ethanol was removed and the final volume was
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c 2009 Society of Chemical Industry
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made up to 25 mL with distilled water. The resulting extract was
passed through a 0.45 µm membrane filter and used for analysis
by high-performance liquid chromatography (HPLC).
For the sugars, 5 mL of extract was passed through a Sep-Pak
C18 cartridge (Waters, Milford, MA, USA) and the eluate was used
for sugar analysis. 20 µL of the eluate was injected into a high-
performance liquid chromatograph with a Zorbax carbohydrate
analytical column (150 × 4.6 mm), a refractive index detector,
and at a flow rate of 1.0 mL min−1 using water as eluent and a
column temperature of 80 ◦ C. Individual sugars were quantified by
comparison with peak areas of individual sugar standards. Results
were expressed as g kg−1 FW. Organic acids were determined
using 5 mL of extract that had been passed through a cation
exchange resin (Dowex 50, Bojie Resin Techology Co., Suzhou,
Jiangsu, China) column with 50 mL water. For organic acids, a
20 µL sample of the eluate was injected into a high-performance
liquid chromatograph with a reverse-phase Zorbax eclipse XDB C18
analytical column (250 × 4.6 mm). The flow rate was 0.8 ml min−1
using 50 mmol L−1 KH2 PO4 as eluent. The column temperature was
30 ◦ C. Organic acid components were detected at a wavelength of
214 nm. Quantification of individual organic acids was made using
peak areas of individual acid standards. Results were expressed as
g kg−1 FW.
Sample preparation for total phenolics, total flavonoids, total
carotenoids and antioxidant capacity
To prepare the fruit extracts, 5 g of flesh tissue from each replicate
was homogenized with 5 mL of 950 mL L−1 cold ethanol and
centrifuged at 10 000 × g for 15 min, and a further 5 mL of 800 mL
L−1 cold ethanol extracted the residua again. The supernatants
were combined to make a final volume of 25 mL. The ethanol
extract was used for analysis of total phenolics, total flavonoids,
total carotenoids and antioxidant capacity.
Analysis of total phenolics, flavonoids and carotenoids
Total phenolics content in loquat fruit extracts was determined
according to the Folin–Ciocalteu procedure,14 and results were
expressed as grams of gallic acid equivalent (GAE) per kilogram
of fresh weight. Total flavonoids content in ethanol extract of
loquat fruit was determined according to the method of Toor and
Savaga,15 the result was expressed as grams of rutin equivalent per
kilogram of fresh weight. Total carotenoids content was measured
spectrophotometrically according to the method of Kichtenthaler
and Wellburn16 and the result was expressed as grams per kilogram
of fresh weight.
Assay for scavenging activity against DPPH, superoxide
and hydroxyl radicals and the reducing power
The DPPH radical scavenging activity of the sample extract
was estimated following the method of Larrauri et al.17 An
aliquot (0.1 mL) of the ethanol extract was added to 2.9 mL of
DPPH (120 µmol L−1 ) in methanol. The absorbance at 517 nm
was measured after the reaction mixtures were incubated for
30 min at 30 ◦ C in the dark. Solvent containing 120 µmol L−1
DPPH without mixing with sample solution was used as control.
The result was calculated according the following formula:
DPPH radical scavenging activity (%) = 100 − (absorbance of
sample/absorbance of control) × 100.
The assay of superoxide scavenging activity was based on
the capacity of the ethanol extract to inhibit formazan for-
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mation by scavenging the superoxide radicals generated in a
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riboflavin–light-nitroblue tetrazolium (NBT) system.18 The total
volume of reactant was 5.0 mL and the mixture contained 50 mmol
L−1 phosphate buffer (pH 7.8), 4 µmol L−1 riboflavin, 0.2 mmol L−1
methionine, 75 µmol L−1 NBT, 3 mmol L−1 ethylenediaminete-
traacetic acid (EDTA) and 0.1 mL ethanol extract. The same reaction
mixture containing 50 mmol L−1 phosphate buffer (pH 7.8) instead
of sample solution was used as control. Absorbance was measured
at 560 nm after the reaction mixture was illuminated at 20 ◦ C under
fluorescent lamp (4000 lx) for 25 min. The percentage inhibition of
superoxide generation was calculated using the following formula:
superoxide radical scavenging activity (%) = 100 − (absorbance
of sample/absorbance of control) × 100.
Hydroxyl radical scavenging activity of the ethanol extract was
determined by the deoxyribose method described previously.19
Five-milliliter reactant mixtures containing 50 mmol L−1 phos-
phate buffer (pH 7.5), 10 mmol L−1 hydrogen peroxide, 5 mmol
L−1 ferric trichloride, 5 mmol L−1 ascorbate, 20 mmol L−1 de-
oxyribose, 1 mmol L−1 EDTA and 0.5 mL of sample solution or
0.5 mL of 50 mmol L−1 phosphate buffer (pH 7.5) as control. The
mixtures were then incubated at 40 ◦ C for 1 h. The reaction was
terminated by adding trichloroacetic acid (50 g L−1 ), followed by
the addition of thiobarbituric acid (2 g L−1 ) and boiled in a water
bath for 15 min. Absorbance of the color was measured at 535 nm.
Decreased absorbance of the mixture indicated scavenging ability
and the result was calculated according the following formula:
hydroxyl radical scavenging activity (%) = 100 − (absorbance of
sample/absorbance of control) × 100.
The reducing power of the ethanol extract was determined
according to the method of Oyaizu.20 A 2.5 mL diluted ethanol
extract of loquat fruit was mixed with 2.5 mL phosphate buffer
(0.2 mol L−1 , pH 6.6) and 2.5 mL potassium ferricyanide (10 g L−1 ).
The mixture was incubated at 50 ◦ C for 20 min. A portion (2.5 mL)
of acetic acid (100 mL L−1 ) was added to the mixture, which was
then centrifuged at 10 000 × g for 10 min. The upper layer of
solution (2.5 mL) was mixed with distilled water (2.5 mL) and ferric
trichloride (0.5 mL, 1 g L−1 ), and the absorbance was measured at
700 nm. Decreased absorbance of the reaction mixture indicated
decreased reducing capacity.
Measurements of PAL and PPO activities
Ten fruit from each treatment replicate were peeled, cut into
small pieces, and the bulked fruit samples (from the 10 fruit)
were frozen in liquid nitrogen and stored at −20 ◦ C until analysis.
PAL was extracted with 0.2 mol L−1 sodium borate buffer at
pH 8.7 containing 20 mmol L−1 of β-mercaptoethanol. For
PPO extraction, 1 g of flesh tissue was ground with 5 mL of
0.2 mol L−1 sodium phosphate buffer (pH 6.5) containing 10 g−1 L
polyvinylpyrrolidone. All extract procedures were conducted at
4 ◦ C. PAL activity was assayed by the method described by
Zucker.21 The assay medium contained 0.1 mL enzyme extract
and 1 mL L-phenylalanine. After incubation at 40 ◦ C for 1 h, the
reaction was stopped by adding 0.2 mL of 6 mol L−1 HCl. One unit
of PAL activity was defined as the amount of enzyme that causes an
increase in absorbance of 0.01 at 290 nm in 1 h under the specified
conditions. PPO activity was assayed following the method of Murr
and Morris.22 The assay medium contained 0.1 mL enzyme extract
and 2 mL catechol. PPO activity was determined by measuring
absorbance at 410 nm. One unit of PPO activity was defined as the
amount of enzyme that caused an increase in absorbance of 0.01
at 410 nm in 1 min under the specified conditions.
Protein content in the enzyme extracts was estimated using the
Bradford23 method, using bovine serum albumin as a standard.
Specific activity of the enzymes was expressed as units per
milligram of protein.
Statistical analysis
Experiments were performed using a completely randomized
design. All statistical analyses of variance were calculated over two
factors: treatment and time in storage with SPSS 13.0 (SPSS Inc.,
Chicago, IL, USA). The main effects were analyzed and the means
were compared by Duncan’s multiple range tests. Differences at
P < 0.05 were considered as significant.
RESULTS AND DISCUSSION
Effects of MeJA treatment on respiration rate and ethylene
production
Loquat fruit is a non-climacteric fruit that exhibits a decrease in
respiration rate and ethylene production during storage. MeJA
treatment was effective in inhibiting respiration rate and ethylene
production (Fig. 1). Respiration rate and C2 H4 production in control
fruit were 145.9% and 53.3% higher than those in MeJA-treated
fruit on the 35th day of storage, respectively. The lower respiration
rate in MeJA-treated fruit may result in lower consumption of
sugars and organic acids and hence better retention of their
contents.
Effect of MeJA treatment on sugar and organic acid contents
Sucrose, fructose and glucose are the main sugars in loquat fruit.
For sucrose content, a steady decrease was observed during

10 0.5
A B
control
8 MeJA 0.4
(ml CO2 kg-1 FW h-1)

C2H4 production
(µl kg-1FW h-1)
Respiratory rate

6 0.3

4 0.2

2 0.1

0 0.0
0 7 14 21 28 35 0 7 14 21 28 35
Storage time (day)

Figure 1. Effect of MeJA on respiration rate (A) and C2 H4 production (B) of loquat fruit during storage at 1 ◦ C. Values are the means ± SE of triplicate
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assays. Vertical bars represent the standard errors of the means.

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300 50
A D
250 40

Malic acid content

Sucrose content

(g kg-1 FW)
200

(g kg-1 FW)
30
150
20
100 control control
MeJA MeJA
50 10

0 0

240 4
B E
200

Oxalic acid content

Fructose content

3
160

(g kg-1 FW)
(g kg-1 FW)

120 2

80
1
40

0 0

260 12
C F
240 10

Lactic acid content

Glucose content

(g kg-1 FW)
220 8
(g kg-1 FW)

200 6

180 4

160 2

140 0
0 7 14 21 28 35 0 7 14 21 28 35
Storage time (day)

Figure 2. Effect of MeJA on contents of sucrose (A), fructose (B), glucose (C), malic (D), oxalic (E) and lactic (F) acid of loquat fruit during storage at 1 ◦ C.
Values are the means ± SE of triplicate assays. Vertical bars represent the standard errors of the means.

the whole storage time, while fructose and glucose contents
increased during the initial storage period and then decreased
at the later storage time (Fig. 2(A)–(C)). Fruit treated with MeJA
tended to maintain significant (P < 0.05) higher levels of sugars
compared to the control fruit. At the end of storage, the contents
of sucrose, fructose and glucose in MeJA-treated fruit were 32.1%,
37.3% and 13.5% higher than that in control after storage for
35 days, respectively. Malic acid is the predominant acid in loquat
fruit; oxalic and lactic acids were also present, but in smaller
concentrations. Malic acid and oxalic acid levels decreased during
the whole storage period; significantly (P < 0.05) higher levels of
these two acids were detected in treated fruit than in control fruit
(Fig. 2(D) and (E)). Lactic acid level increased during the first 2 and
3 weeks of storage and then declined in control and MeJA-treated
fruit, respectively (Fig. 2(F)). The levels of these three acids
remained significantly (P < 0.05) higher in MeJA-treated fruit
than in the control fruit, especially at the end of the storage. The
content of malic, oxalic and lactic acid in MeJA-treated fruit were
22.0%, 150.0% and 103.4% higher than that in control fruit on the
35th day of storage, respectively. These results suggest that MeJA
treatment can improve the organoleptic quality of loquat fruit
during storage by maintaining higher levels of sugars and organic
been shown to maintained higher levels of sugars and organic
acids in a number of horticultural crops, including fresh-cut
kiwifruit, radish, raspberry and zucchini squash.9,10,24 – 26 Since
sugars and organic acids are the main substrates of respiratory
metabolism, the maintenance of their levels by MeJA could be
due to the inhibition of fruit respiration (Fig. 1(A)).
Effect of MeJA treatment on contents of total phenolics,
flavonoids and carotenoids
Total phenolics content in both control and MeJA-treated fruit
decreased during storage; MeJA treatment markedly inhibited the
decline of total phenolics, thus maintaining significant (P < 0.05)
higher levels of phenolics throughout the experimental period
compared to the control fruit. As shown in Table 1, total phenolics
content in treated fruit was 26.3% higher than that in control
fruit on the 35th day of storage. The total flavonoids content in
control and MeJA-treated fruit increased during the first 14 and
21 days of storage, respectively, and decreased gradually with
prolonged storage time. A significantly (P < 0.05) higher level of
total flavonoids was observed in MeJA-treated fruit throughout the
whole storage period compared with control. The level in MeJA-
treated fruit was 45.1% higher than that in control fruit after 35 days
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acids. MeJA treatment, applied either pre- or postharvest, has also of cold storage (Table 1). Total carotenoids content increased

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Effects of MeJA treatment on activities of PAL and PPO

Table 1. Effect of MeJA on contents of total phenolics, flavonoids
and carotenoids of loquat fruit during storage at 1 ◦ Ca The metabolism of phenolic compounds is regulated by the activity
of various enzymes. The first step necessary for the synthesis of the
Total phenolics Total flavonoids Total carotenoids
phenylpropanoid skeleton in higher plants is the deamination of L-
Treatment (g kg−1 FW) (g kg−1 FW) (g kg−1 FW)
phenylalanine, giving rise to trans-cinnamic acid and ammonium.
Day 0 1529.1 ± 11.9a 81.2 ± 1.2bc 7.0 ± 1.9c This reaction is catalyzed by the enzyme PAL, which is commonly
Day 7 considered the principal enzyme in the biosynthesis of phenolic
Control 1230.8 ± 45.8c 59.9 ± 1.1de 8.1 ± 0.5bc compounds.32 In addition, the metabolism of phenolic compounds
MeJA 1349.9 ± 0.0bc 92.4 ± 0.9a 8.9 ± 0.0abc also includes the action of oxidative enzymes such as PPO, which
catalyzes the oxidation of phenols to quinones.33 The activities
Day 14
of PAL and PPO were therefore examined with the aim of better
Control 1090.5 ± 17.7d 54.8 ± 0.2de 9.0 ± 0.7abc understanding the process involved in phenolic metabolism in
MeJA 1366.4 ± 18.2b 85.6 ± 0.6ab 8.2 ± 0.2bc loquat fruit after MeJA treatment. As shown in Fig. 3, activities of
Day 21 PAL and PPO in loquat fruit increased with storage time. MeJA
Control 727.0 ± 44.4fg 61.6 ± 1.4d 8.8 ± 0.4abc treatment significantly (P < 0.05) inhibited the increase of both
MeJA 1250.0 ± 5.4bc 87.1 ± 0.6ab 8.4 ± 0.5abc enzymes. At the end of storage, the activities of PAL and PPO
in MeJA-treated fruit were 35.6% and 24.1% lower than that in
Day 28
control after storage for 35 days, respectively. It was reported that
Control 611.9 ± 31.0g 58.1 ± 0.0de 9.4 ± 1.0ab MeJA treatment induced accumulation of phenolic compounds,
MeJA 890.0 ± 88.9e 88.1 ± 1.2ab 8.1 ± 0.8bc such as caffeic acid derivative, in romaine lettuce, possibly through
Day 35 the action of PAL.34 However, in the present study, treatment with
MeJA significantly (P < 0.05) decreased PAL activity (Fig. 3(A))
Control 645.9 ± 71.2g 51.3 ± 0.8e 10.6 ± 1.1ab
while still maintaining higher phenolic compounds (Table 1),
MeJA 813.0 ± 49.3ef 74.9 ± 6.6c 9.5 ± 0.9ab
which was completely different from the previous report. Ossipov
a Data expressed as mean ± SE of triplicate assays. Different letters in
et al.35 suggested that, apart from the synthesis of phenolic
the same column indicate statistically significant difference at P < 0.05. compounds via PAL activation, some phenolic compounds such
as gallic acid or derivatives could also be synthesized through
other means such as the shikimate pathway. Therefore, whether
the shikimate pathway was involved in the phenolic metabolism
gradually with storage time, but no significant differences in in loquat fruit deserved further study. During storage, PPO activity
total carotenoids content were found between control and MeJA- in MeJA-treated fruit was lower than that in control fruit. This
treated fruit during storage (Table 1). negative shift of PPO activity provoked by MeJA could be due to a
As an endogenous phytohormone, MeJA plays a key role in conformational change of the enzyme or to a modification of the
regulating a great diversity of physiological and biochemical active site, namely a reduction in the cupric ion of the enzyme.36
processes in plants, including stimulating the biosynthesis of As PPO is necessary to initiate phenol oxidation to dark-brown
secondary metabolites.8 MeJA has been shown to induce stilbene melanin,33 it is possible that the observed increase of PPO activity
accumulation in leaves and berries of grapevine plants, increase during the whole storage time might explain the decrease of total
the accumulation of anthocyanins and phenolics in apples, phenolic content. These results confirmed the beneficial effect of
raspberries, strawberries and blackberries, and promote β- MeJA in delaying the oxidation of phenolic compounds and thus
carotene synthesis in apple peel.22,27 – 31 In the present study, preserving higher total phenolic content.
loquat fruit treated with MeJA exhibited significantly (P < 0.05)
higher levels of total phenolics and flavonoids compared to Effect of MeJA treatment on antioxidant activity
the control fruit, which suggests that MeJA may improve the Reactive oxygen species (ROS), such as superoxide radical,
antioxidant status of the fruit by positively affecting phenolic hydroxyl radical and hydrogen peroxide, are by-products of
metabolism. normal metabolism and generated when metabolism interacts

8 160
PPO activity (U mg-1 protein)
PAL activity (U mg-1 protein)

A B
control
6 MeJA 120

4 80

2 40

0 0
0 7 14 21 28 35 0 7 14 21 28 35
Storage time (day)

Figure 3. Effect of MeJA on activities of PAL (A) and PPO (B) of loquat fruit during storage at 1 ◦ C. Values are the means ± SE of triplicate assays. Vertical
2068

bars represent the standard errors of the means.

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Table 2. Effect of MeJA on scavenging activity against DPPH, superoxide and hydroxyl radicals, and reducing power of loquat fruit during storage
at 1 ◦ Ca

Treatment DPPH scavenging (%) Superoxide scavenging (%) Hydroxyl scavenging (%) Reducing power

Day 0 40.7 ± 1.8a 48.0 ± 3.2e 84.9 ± 0.3a 0.209 ± 0.005a

Day 7
Control 28.3 ± 0.5bcd 58.3 ± 1.4cd 71.6 ± 0.3b 0.151 ± 0.007cd
MeJA 31.4 ± 0.2b 64.4 ± 2.5ab 68.9 ± 0.4bc 0.186 ± 0.017ab
Day 14
Control 26.4 ± 0.3d 59.9 ± 1.0bc 59.7 ± 0.0de 0.144 ± 0.013de
MeJA 30.0 ± 0.1bc 65.5 ± 0.9ab 69.6 ± 0.7bc 0.182 ± 0.005ab
Day 21
Control 18.5 ± 0.3e 56.7 ± 0.8cd 53.1 ± 0.3fg 0.117 ± 0.017e
MeJA 26.7 ± 2.0d 66.0 ± 0.0a 59.6 ± 0.3de 0.187 ± 0.0ab
Day 28
Control 19.7 ± 0.4e 53.8 ± 3.0de 51.0 ± 1.9g 0.084 ± 0.007f
MeJA 25.7 ± 0.6d 60.0 ± 0.3bc 62.8 ± 0.7d 0.173 ± 0.002bc
Day 35
Control 21.3 ± 1.3e 53.5 ± 2.0de 55.6 ± 3.4ef 0.078 ± 0.005f
MeJA 28.1 ± 1.8cd 60.7 ± 1.6abc 67.1 ± 1.0c 0.187 ± 0.007ab
a
Data expressed as mean ± SE of triplicate assays. Different letters in the same column indicate statistically significant difference at P < 0.05.

with oxygen. Damage mediated by ROS has generally been
considered to be linked with many chronic health problems.37 The
antioxidant constituents in plants, which can quench ROS, have
health-promoting effects in the prevention of chronic problems
and are raising interest among scientists and consumers.2,3 In order
to evaluate the antioxidant activity of biological samples, several
methods have been developed in recent years. We evaluated
the antioxidant activity of loquat fruit treated with MeJA by
the scavenging activity against DPPH, superoxide and hydroxyl
radicals, and the reducing power test in this study. The DPPH
radical scavenging activity decreased during storage, fruit treated
with MeJA exhibited a significantly (P < 0.05) higher DPPH radical
scavenging activity than that in control fruit throughout storage
(Table 2). At the end of storage, the activity in MeJA-treated
fruit was 32.8% higher than that in control after storage for
35 days. A significant positive relationship (R2 = 0.906) between
total phenolic content and DPPH radical scavenging activity was
observed in this investigation, thus the higher DPPH radical
scavenging activity in MeJA-treated loquat fruit in the present
study could be attributed mainly to its higher level of total
phenolics.
Among the ROS, the superoxide and hydroxyl radicals are
the most reactive and induce severe damage to adjacent
biomolecules.37 Loquat fruit extract was found to be a powerful
scavenger of these two radicals. The superoxide radical scavenging
activity of loquat fruit showed a steady increase during the
first 21 days of storage, and then decreased slightly during
the remaining time. Fruit treated with MeJA had a significantly
(P < 0.05) higher superoxide radical scavenging activity during
storage (Table 2). The hydroxyl radical scavenging activity in loquat
fruit decreased with storage time. Fruit treated with MeJA tended
to maintain significantly (P < 0.05) higher scavenging activity of
hydroxyl radical after 7 days of storage compared to the control
radicals in treated fruit were 13.4% and 21.6% higher than that in
control fruit on the 35th day of storage, respectively.
The reducing power of these compounds indicates that they
are electron donors and can reduce the oxidized intermediates of
lipid peroxidation processes, so that they can act as primary and
secondary antioxidants. Previous studies have shown a correlation
between antioxidant activity and reducing power of certain plant
extracts.20 As can be seen in Table 2, the reducing power in
control loquat fruit decreased during storage. Treatment with
MeJA markedly inhibited the decline of reducing power and thus
maintained significantly (P < 0.05) higher reducing power value
than the control fruit during the whole storage period. At the end
of storage, the reducing power in MeJA-treated fruit was 137.5%
higher than that in control after storage for 35 days.
It has been reported that the harmful action of ROS can be
blocked by antioxidant substances which scavenge the free
radicals and detoxify the organism. Plants with high levels of
antioxidants, either constitutive or induced, have been regarded
as having greater resistance to oxidative damage to lipids, proteins
and nucleic acids, thus efficient antioxidant activity is essential in
order to maintain the ROS at relatively low levels.38 MeJA has
been demonstrated to be effective in enhancing antioxidant
activity in berry fruits such as raspberries, strawberries and
blackberries.11,12,25,29,30 In the present study, the scavenging
activity against DPPH, superoxide and hydroxyl radicals, and the
reducing power in MeJA-treated fruit, were significantly higher
than those in control fruit. This result suggests that a postharvest
application of MeJA will improve the health benefit of loquat fruit
by enhancing the antioxidant activity.
In summary, the data presented in this study indicate that a
postharvest application of MeJA positively affected antioxidant
levels, antioxidant activity and overall quality of loquat fruit. MeJA
might exert its effect on increasing phenolics content in loquat
fruit by inhibiting PPO activity. The increased contents of phenolics
2069

fruit (Table 2). The scavenging activities of superoxide and hydroxyl and flavonoids, elicited by MeJA treatment, resulted in enhanced

J Sci Food Agric 2009; 89: 2064–2070

c 2009 Society of Chemical Industry www.interscience.wiley.com/jsfa
 www.soci.org
antioxidant activity. Thus MeJA has a potential application in
postharvest treatment for improving the functional properties
and maintaining a high-quality product in harvested loquat fruit.
ACKNOWLEDGMENTS
This study was supported by the National Natural Science
Foundation of China (30671462), the Qing Lan Project and
National Scientific and Technical Supporting Program of China
(2006BAD30B03).
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