Subscriber access provided by Nottingham Trent University
Omics Technologies Applied to Agriculture and Food
MeJA promotes phospholipid remodeling and jasmonic acid signaling to alleviate chilling injury in peach fruit Mengshuang Chen, Huimin Guo, Shuqi Chen, Tingting Li, Meiqing Li, Arif Rashid, Changjie Xu, and Ke Wang J. Agric. Food Chem., Just Accepted Manuscript • DOI: 10.1021/acs.jafc.9b03853 • Publication Date (Web): 16 Aug 2019 Downloaded from pubs.acs.org on August 19, 2019
Just Accepted “Just Accepted” manuscripts have been peer-reviewed and accepted for publication. They are posted online prior to technical editing, formatting for publication and author proofing. The American Chemical Society provides “Just Accepted” as a service to the research community to expedite the dissemination of scientific material as soon as possible after acceptance. “Just Accepted” manuscripts appear in full in PDF format accompanied by an HTML abstract. “Just Accepted” manuscripts have been fully peer reviewed, but should not be considered the official version of record. They are citable by the Digital Object Identifier (DOI®). “Just Accepted” is an optional service offered to authors. Therefore, the “Just Accepted” Web site may not include all articles that will be published in the journal. After a manuscript is technically edited and formatted, it will be removed from the “Just Accepted” Web site and published as an ASAP article. Note that technical editing may introduce minor changes to the manuscript text and/or graphics which could affect content, and all legal disclaimers and ethical guidelines that apply to the journal pertain. ACS cannot be held responsible for errors or consequences arising from the use of information contained in these “Just Accepted” manuscripts.
is published by the American Chemical Society. 1155 Sixteenth Street N.W., Washington, DC 20036 Published by American Chemical Society. Copyright © American Chemical Society. However, no copyright claim is made to original U.S. Government works, or works produced by employees of any Commonwealth realm Crown government in the course of their duties.
Page 1 of 39
Journal of Agricultural and Food Chemistry
1
MeJA promotes phospholipid remodeling and jasmonic acid
2
signaling to alleviate chilling injury in peach fruit
3 4
Mengshuang Chen1, Huimin Guo2, Shuqi Chen1, Tingting Li1, Meiqing Li1, Arif
5
Rashid3, Changjie Xu4, Ke Wang1*
6 7
1
8
University, Hefei 230036, China
9
2
Anhui Engineering Laboratory for Agro-products Processing, Anhui Agricultural
Center for Biological Technology, Anhui Agricultural University, Hefei 230036,
10
China
11
3
School of Life Science, Anhui Agricultural University, Hefei 230036, China
12
4
College of Agriculture and Biotechnology/Zhejiang Provincial Key Laboratory of
13
Horticultural Plant Integrative Biology, Zhejiang University, Zijingang Campus,
14
Hangzhou 310058, China
15 16 17
* Corresponding author:
18
Dr. Ke Wang (E-mail:
[email protected])
19
ACS Paragon Plus Environment
Journal of Agricultural and Food Chemistry
20
ABSTRACT
21
Chilling injury (CI) is a physiological disorder induced by cold, which heavily limit
22
crops production and postharvest preservation worldwide. MeJA can alleviate CI in
23
various fruit species, including peach; however underlying molecular mechanism is
24
poorly understood. Here, changes in contents of phenolics, lipids, and jasmonic acid
25
(JA), and gene expressions, are compared between MeJA and control fruit. Exogenous
26
MeJA inhibited expressions of PpPAL1, PpPPO1 and PpPOD1/2, but did not affect
27
phenolics content. Furthermore, MeJA fruit showed lower relative electrolyte leakage,
28
indicating less membrane damage. Meanwhile, the enrichment of linoleic acid in the
29
potential lipid biomarkers, especially PC, PE and PG, coincided with lower
30
expressions of PpFAD8.1, but higher PpLOX3.1, and JA content. In the JA signaling
31
pathway, MeJA significantly upregulated expressions of PpMYC2.2 and PpCBF3, but
32
downregulated PpMYC2.1. In conclusion, adjustments of fatty acids in phospholipids
33
contribute to MeJA-induced alleviation of CI in peach fruit, via induction of JA
34
mediated CBF pathway.
35 36 37 38
KEYWORDS: Peach, MeJA, chilling injury, fatty acids, lipid
39
ACS Paragon Plus Environment
Page 2 of 39
Page 3 of 39
Journal of Agricultural and Food Chemistry
40
INTRODUCTION
41
For horticultural products of typical crop categories, cold storage is often used to
42
prolong their postharvest life, whereas long-term storage will cause a series of
43
physiological disorders termed chilling injury (CI), especially for certain tropical and
44
subtropical fruit and vegetables 1. Therefore, the elucidation for CI is of great
45
economic importance and has become a hot scientific issue to be addressed globally
46
during recent decades. Similar microscopic changes were observed during CI of
47
different horticultural products, while macroscopic changes were quite different, such
48
as abnormal skin colors, woolliness 2 and lignin development 3-5, surface and internal
49
browning (IB) 6-10.
50
IB is one of classic CI symptoms, and has often been considered as a visible
51
maker to reflect the CI intensity 11. The occurrence of IB is mainly facilitated by three
52
factors, including phenolic substrates, relative enzymes, including phenylalanine
53
ammonialyase (PAL), polyphenol oxidase (PPO) and peroxidase (POD), as well as
54
their compartmentalization by membrane as barriers 12. The function and integrity are
55
crucial for the compartmentalization of membrane, and it has been reported that
56
membrane lipids have a close impact on the function and integrity of the membranes.
57
Based on their structure and content difference, membrane lipids are generally divided
58
into glycerolipids and sphingolipids. The unsaturated level of total fatty acids (FAs)
59
was found to be positively correlated with chilling tolerance of many fruit species
60
under cold conditions 13-16. In addition, based on lipidome technique, the types of lipid
61
molecular species associated with chilling tolerance have recently been identified in
62
the plants
63
and fatty acids (FAs) have been identified, such as FA desaturase (FAD), and
17-19
and fruit
7, 20
. Meanwhile, many enzymes leading to changes to lipids
ACS Paragon Plus Environment
Journal of Agricultural and Food Chemistry
Page 4 of 39
64
lipoxygenase (LOX), involved in desaturation of FAs and peroxidation, respectively.
65
It is reported that these two enzymes are closely associated with CI 21-23.
66
Among diverse strategies against CI, phytohormone treatments often confer
67
effects of CI alleviation and quality on extensive range of horticultural products,
68
including ethylene
69
brassinosteroids 29-30, salicylic acid
70
that CI was ameliorated by JA exogenous donor MeJA in kinds of horticultural
71
products, including loquat fruit 34-35, orange fruit 36, pomegranate fruit 37, tomato fruit
72
22, 38-39
73
significant CI tolerance in diverse horticultural products, its economic and applicable
74
characteristics make it become a promising postharvest strategy for their quality
75
control.
24
, abscisic acid 31-33
25
, gibberellin acid
26-27
, melatonin
22, 28
,
, and jasmonic acid (JA). It has been reported
, papaya fruit 40-41, banana peel 41, kiwifruit 42, peach fruit 43-45. In addition to its
76
The mechanism of MeJA conferring CI resistance has been extensively studied. It
77
has been reported that JA biosynthesis and its signaling transduction pathway have an
78
effect on chilling tolerance
79
from release of unsaturated FAs from membrane lipids and further catalyzed by a
80
series of enzymes including LOX, allene oxide synthase, allene oxide cyclase,
81
cis-(+)-12-oxo-phyto-dienoic acid reductase. In addition, antioxidant systems
82
shock proteins 46, crytoprotectants 38, 44, energy substances and membrane lipids 43, are
83
reported to be part of effects of MeJA on chilling tolerance. Furthermore, the cascades
84
of C-repeat-binding factors (CBFs) couple with its inducer of CBF expression (ICE),
85
has been proved to be linked with the response induced by MeJA through interaction
86
between their corresponding key transcription factor ICE1 and MYC2 in banana fruit
87
41
39, 41
. The biosynthesis of the endogenous JA originates
.
ACS Paragon Plus Environment
35
, heat
Page 5 of 39
Journal of Agricultural and Food Chemistry
88
Although the understanding of chilling tolerance induced by MeJA has been
89
greatly expanded in past decades, the mechanism of MeJA on IB alleviation shared by
90
many horticultural products is still controversial. For phenolic content, some studies
91
showed negative effects of MeJA 37, 45, whereas others were positive
92
membrane barrier, MeJA could inhibit membrane damage, and maintain a high ratio
93
between unsaturated FAs and saturated FAs 43, however, the details of membrane lipid
94
reprogramming, and key genes remain to be elucidated. Peach is a typical climacteric
95
fruit with short shelf life of often less than one week. Cold storage can cause a variety
96
of CI symptoms such as woolliness, loss of juice, volatiles and ability to ripen, as well
97
as IB. It has been reported that peach IB can be alleviated by MeJA, and phenolics
98
metabolism has been investigated at physical and biochemical level, however
99
underlying molecular mechanism is not fully understood, and lipidome adjustments
100
47-48
. In term of
by MeJA remains to be uncovered.
101
‘Xiazhimeng’ is among stony-hard peaches and shows high chilling sensitiveness
102
according to our preliminary studies. This study aims to provide more insights into
103
lipid reprogramming involved in CI alleviated by MeJA. The results are helpful to
104
understand the cold tolerance mechanism conferred by MeJA, and the technological
105
innovation of CI prevention and amelioration for horticultural products.
106 107
MATERIALS AND METHODS
108
Plant Materials and Treatments
109
Peach (Prunus persica Batsch cv. Xiazhimeng) fruit were harvested at commercial
110
maturity from an orchard in Hefei, Anhui, China, and were transferred to the
111
laboratory once harvest. The uniform fruit without visual defects were randomly
112
divided into two groups. One group of fruit were fumigated among 10 μmol L-1 MeJA
ACS Paragon Plus Environment
Journal of Agricultural and Food Chemistry
Page 6 of 39
113
(Sigma-Aldrich, St. Louis, MO, USA) for 24 h at 20℃. After ventilation for 1 h, the
114
fruit were stored at 0℃; the other group of fruit were also held under the same
115
condition as the first group without MeJA fumigation. After 63 days of cold storage,
116
the fruit were transferred to 20℃ for six days to mimic shelf life. The samplings were
117
performed at the end point of shelf life (63+6d). Fifteen fruit were sampled at the
118
sampling point and randomly divided into three biological replicates with five fruit in
119
each replicate. The mesocarp was sliced and immediately frozen in liquid nitrogen
120
and stored at −80°C for further analysis.
121
Internal browning index and electrolyte leakage rate
122
An IB index was calculated to evaluate the degree of flesh browning based on a
123
previous study 7. Electrolyte leakage rate was determined as described by Wang 49.
124
Transcriptomic analysis and real-time quantitative PCR
125
The RNA samples of peach mesocarp were sent to commercial company (Personal
126
Biotechnology Co., Ltd, Shanghai City, China) for transcriptome sequencing. Briefly,
127
total RNA of mesocarp tissues of peach fruit were extracted using kit of TRIzol
128
(Ambion, Austin, USA). Reads were filtrated by three steps of removing adaptor
129
sequences, low quality (< Q20) and length shorter than 50bp, followed by quality
130
checking with FastQC (version 0.11.6). The clean reads were then aligned to the
131
peach genome (V2.1) by bowtie/tophat2. Gene-level raw read counts were normalized
132
using transcript per million (TPM) by Stringtie (version 1.33).
133
Real-time quantitative PCR was performed as described by Wang 49. The primers
134
information is listed in the Table S1.
135
Lipid extraction
136 137
Lipid extraction was conducted followed by the reported method
20
with minor
modifications. Mesocarp of peach (n = 5) were ground to power under liquid nitrogen,
ACS Paragon Plus Environment
Page 7 of 39
Journal of Agricultural and Food Chemistry
138
and 25 mg of powder were weighed for further lipid extraction. 1 mL of methanol:
139
methyl tertbutyl ether: water (1:3:1) mixtures were prepared and mixed with the
140
sample for 20 min at 4℃; after addition of water: methanol (3:1) mixture, lipophilic
141
phase were collected, and dried with high purity nitrogen. The dried lipid extracts was
142
recovered in buffer B (in the section of lipidome analysis) and further purified
143
through 0.22 μm syringe filter.
144
Lipidome Analysis
145
One microliter of samples was injected on a Waters ACQUITY UPLC BEH C8
146
column (100 mm × 2.1 mm, 1.7 μm, Waters, USA) connected with guard column, on
147
platform of a Thermo Scientific UHPLC system and coupled with Q Exactive mass
148
spectrometer (Thermo Fisher Scientific, Bremen, Germany). The two mobile phases
149
were water with 0.1% acetic acid (phase A), and acetonitrile with 0.1% acetic acid
150
(phase B), respectively. The gradient separation conditions were as follows: flow rate
151
of 0.3 mL min-1. 1 min phase A, and then 5 min linear gradient from 100% to 50%
152
before 24 min linear gradient from 50% to 0%. The phase A was recovered to 100%
153
and re-equilibrated for 4 min before next injection.
154
The mass analysis was conducted under both positive and negative ion modes. For
155
positive ion mode, the full MS scan type range from 70 m/z to 1050 m/z; resolution
156
70000, sheath gas flow rate 45, aux gas flow rate 15 arb, sweep gas flow rate 1 arb,
157
spray voltage 3.8 KV, capillary temperature 350℃, S-lens RF level of 60, aux gas
158
heater temperature 350℃; and for the negative ion mode, full MS scan type range
159
from 100 m/z to 1500 m/z; resolution 70000, sheath gas flow rate 40 arb, aux gas flow
160
rate 10 arb, sweep gas flow rate 0, spray voltage 3.1 KV, capillary temperature 320℃,
161
S-lens RF level of 60, aux gas heater temperature 350℃.
162
Chromatograms peak detection, integration and alignment across samples were
ACS Paragon Plus Environment
Journal of Agricultural and Food Chemistry
163
performed using LipidsearchTM software (version 4.1, Thermo Fisher Scientific,
164
Germany).
165
Jasmonic acid determination
166
Two hundred micrograms of sample powder were weighed for JA extraction. The
167
1.5 mL extraction buffer (methanol: water: formic acid = 7.9: 2: 0.1) was mixed with
168
the sample. After ultrasonication for 30 min on ice, the mixtures were incubated at 4℃
169
for 12 h, and centrifuged (13000×g) for 20 min at 4℃. The supernatants were
170
decanted to new tubes and the remaining fruit tissue was retracted followed by above
171
manipulations. The twice supernatants were collected with first one, and enriched
172
through MAX SPE column (60mg, 3mL, Waters, USA). The final elution was dried to
173
powders with high purity nitrogen and recovered with solvent (acetonitrile: formic
174
acid: water = 5: 0.1: 94.9).
175
JA detection were performed on ACQUITY UPLC H-Class platform (Waters,
176
USA) through Waters ACQUITY UPLC HSS T3(100 * 2.1 mm, 1.7 μm, Waters,
177
USA), with 1.0 μl volume of injection. Two-phase wash buffer consisted of phase A
178
(0.1% formic acid in water) and phase B (0.1% formic acid in acetonitrile). Gradient
179
separation program was as following: after equilibrium at 95% phase A, 1 min linear
180
gradient from 95% to 30%, then constant for 1 min; recovered to 95% and kept
181
equilibrium for 1 min before next injection. MassLynx software (version 4.1, Waters,
182
USA) was used for signal extraction and quantification.
183
Statistics analysis
184
The Experiment was designed based on random principles; variance analysis via
185
Student T-test method was conducted with Microsoft Excel and OPLS-DA was
186
performed by ‘ropls’ package on R.
187
ACS Paragon Plus Environment
Page 8 of 39
Page 9 of 39
Journal of Agricultural and Food Chemistry
188
RESULTS
189
Effect of MeJA on internal browning and phenolic metabolism of peach fruit
190
IB index and relative electrolyte leakage rate of MeJA-treated fruit was
191
significantly 50% and 20% lower, respectively, than that of control fruit at the end of
192
shelf life (63 days of cold storage at 0℃ plus 6 days at 20℃, Fig. 1), suggesting that
193
MeJA could significantly inhibit CI-induced IB and cellular membrane permeability
194
in ‘Xiazhimeng’ peach fruit.
195
In order to examine expression of genes related to IB, transcriptomic data in the
196
samples was obtained and validated by real-time quantitative PCR (Fig S1). In
197
comparison with control fruit, significantly lower transcript levels of PpPPO1 and
198
PpPOD1/2 involved in phenolic depletion were observed in MeJA fruit (Fig 2A).
199
Similarly, PpPAL1 involved in phenolic biosynthesis exhibited significantly lower
200
transcript levels. However, no significant differences were observed in the content of
201
total phenolics between MeJA and control fruit (Fig S2).
202 203
Untargeted lipidome reprogramming in peach fruit
204
Since the changes of phenolics cannot fully explain the less IB in MeJA fruit,
205
whereas membrane permeability was significantly inhibited by MeJA, further
206
membrane lipid reprogramming was evaluated. In order to investigate adjustments of
207
membrane lipids, an untargeted lipidomic analysis platform for peach fruit was
208
developed, and 56 lipid species were identified (Table. 1). Phosphatidylcholines (PCs,
209
nine species) and phosphatidylethanolamines (PEs, nine species) in phospholipids
210
were the most abundant lipid species after alignment across samples in this study. In
211
addition, sphingolipids including glycosylceramide (GCer) and ceramide (Cer) were
ACS Paragon Plus Environment
Journal of Agricultural and Food Chemistry
212
identified. These findings indicate that our system is suitable for the investigation of
213
polar glycerolipids and partial sphingolipids.
214
Several lipid species including two PCs and three PEs, and two (PGs), one Cer
215
and one (TG) were found to be significantly differentially accumulated between
216
MeJA and control fruit (Table. 1). The contents of total PEs and PGs, showed higher
217
levels in MeJA fruit relative to control fruit (Fig 3A). Although no considerable
218
changes in levels of total PCs, two species of PCs showed significant difference in
219
MeJA relative to control, including PC (18:2/18:2), PC (16:0/18:2) with higher levels
220
(Fig 3B). Besides, PE (16:0/18:2), PE (18:0/18:2), PE (18:2/18:2), PG (16:0/18:1),
221
PG (16:0/18:2) and PI (16:0/18:2), exhibited higher accumulation, as well.
222
In order to filter out lipid species contributing to separation of MeJA fruit from
223
control fruit, an orthogonal partial least squares-discriminant analysis (OPLS-DA)
224
model was established. It is well fitted (R2X = 0.935) and predictively (Q2Y = 0.975),
225
and there is clear divergence between MeJA and control fruit in the OPLS-DA score
226
plot (Fig 4A). Variable importance of projection (VIP) represents a summary vector
227
that explains the total importance of variances in the model. A clear negative
228
correlation between VIP and Student t-test P-value was observed, and three PCs
229
(18:0/18:2, 18:2/18:2, 16:0/18:2), two PEs (18:0/18:2, 18:2/18:2), PG (16:0/18:1), Cer
230
(d16:0/2:0) and FA (20:1) were of both high VIP (> 1) and low P-value (< 0.05, Fig
231
4B), indicating potential biomarkers for MeJA fruit during CI development after
232
long-term cold storage.
233 234
Changes of unsaturation status in fatty acid acyls in differentially accumulated
235
lipid molecular species
ACS Paragon Plus Environment
Page 10 of 39
Page 11 of 39
Journal of Agricultural and Food Chemistry
236
Intriguingly, with exception of linolenic acid (18:3), all FA acyl chains especially
237
linoleic acid (18:2) in potential phospholipid biomarkers, especially PC, PE and PG,
238
showed higher levels (approximately two-fold higher) in MeJA fruit, compared with
239
control fruit (Fig 4C), indicating significant effects of MeJA on unsaturation of FA in
240
phospholipids. Therefore, transcript encoding enzymes involved in FA metabolism
241
were studied based on transcriptomic data. Five genes encoding FAD and three ones
242
encoding LOX displayed differential expression between MeJA fruit and control fruit
243
(Fig 5, 6). Although no significant differences in transcript levels of PpFAD2 in MeJA
244
fruit, significantly lower transcript levels of PpFAD6 and PpFAD8s were observed
245
relative to control fruit, especially PpFAD8.1 (Prupe.6G056100) being approximately
246
4.7-fold lower (Fig 5A). Additionally, PpFAD8.1 presented second highest mRNA
247
abundance among PpFADs. One putative PpLOX3 (Prupe.4g047800, designated
248
PpLOX3.1) in MeJA fruit accumulated 10-fold higher transcript levels than control
249
fruit, whereas PpLOX1, lower (Fig 5B).
250
Unsaturated FAs substrate for the JA biosynthesis, and hence JA level was also
251
detected, and MeJA fruit exhibited significantly 46% higher (p=0.013) levels of that
252
relative to control fruit (Fig 6A). The downstream effector of JA signaling, MYC2,
253
putatively encoded by PpMYC2.1 and PpMYC2.2, exhibited significantly 0.6-fold
254
lower (p=0.036) and 7.3-fold higher (p=0.038) transcript levels in MeJA fruit
255
compared with control fruit (Fig 6B-C).
256
To find associations between JA signals and CBFs, which were considered as key
257
factors in acquirement of chilling tolerance in plants, the differentially expressed
258
PpCBFs were examined. One putative PpCBF3 (Prupe.2G289500) showed
259
significantly 13.38-fold higher (p=0.0063) transcript levels in MeJA fruit relative to
260
control fruit (Fig 6D).
ACS Paragon Plus Environment
Journal of Agricultural and Food Chemistry
261
Page 12 of 39
DISCUSSION
262
Recently IB in peach fruit has involved phenolics metabolism and membrane lipid
263
reprogramming; however a more holistic understanding of mechanism related to its
264
development and tolerance is still limited. In recent years, an approach of lipidome
265
combined transcriptomic analysis, has been performed and provides new insights into
266
this CI symptom
267
reprogramming and phenolic metabolism existed in IB alleviation by low temperature
268
conditioning 7. However, these findings need to be further validated with other
269
strategies such as MeJA, which showed remarkable amelioration of IB in diverse fruit
270
species, such as banana peel, peach, loquat and mango fruit
271
mechanism of MeJA-induced chilling tolerance, many studies focused on its effects
272
on fruit under cold condition, however less attention was paid on CI of fruit after cold
273
removal. In this study, IB alleviation induced by MeJA were observed in peach fruit at
274
the end of shelf life after cold storage for 63 days (Fig 1A); we further confirm that
275
both phenolics metabolism and lipid reprogramming events are involved in CI
276
alleviation processes, and provide more details to understanding on CI.
7, 18, 20
. According to our previous findings, both membrane lipid
35, 40-41, 48, 50
. As for the
277
Here, we propose a model of IB alleviation by MeJA in peach fruit (Fig 7).
278
Inhibition of phenolics metabolism also contributes to alleviation of IB by MeJA,
279
through down-regulating expression of PpPPO1, PpPOD1/2 and PpPAL1. Membrane
280
damage induced by low temperature, such as 0℃, should be alleviated by both
281
reprogramming of lipid molecular species and activation of chilling tolerance
282
pathways. For lipid reprogramming, MeJA inhibits desaturation of linoleic acid (18:2)
283
through downregulating expression of PpFAD8.1. For chilling tolerance pathway,
284
PpCBF3 in CBF signaling pathway is activated by MeJA with enhancement of
ACS Paragon Plus Environment
Page 13 of 39
Journal of Agricultural and Food Chemistry
285
accumulation of JA mediated by up-regulating expression of PpMYC2.2 and
286
PpLOX3.1, and down-regulating that of PpMYC2.1.
287 288
MeJA inhibits both biosynthesis and depletion of phenolics
289
Cold induced IB mainly depends on browning substance of phenolics, which is
290
controlled by generation of phenolics by PAL, and depletion by combined reactions of
291
POD and PPO 28, 51. In our results, MeJA fruit showed less intensity of IB, which is
292
accompanied by a lower level of phenolic metabolism indicated by lower transcript
293
levels of PpPPO1, PpPOD1/2, and PpPAL1 (Fig 2). The reduction of both
294
biosynthesis and depletion could explain the fact of no significant differences of
295
phenolic content in MeJA fruit, in the comparison with control fruit (Fig S2). The
296
results are different to the observation in ‘Baifeng’ peach fruit that the increase in
297
phenolic content and activities of PAL were observed in fruit during shelf life after
298
cold storage
299
‘Baifeng’ is a melting peach, and ‘Xiazhimeng’, stony-hard type. Indeed, dynamics of
300
phenolics contents is hardly consistent among different fruit species when subjected
301
to CI 29, 53.
52
. The cultivar variations should be considered as one of reasons, as
302 303
Promotion of linoleic acid (18:2) in phospholipids against membrane damage
304
In addition to substrate phenolics and enzymes, membrane damage is another
305
factor affecting browning occurrence of horticultural products caused by cold stress,
306
and it depends on the types of lipid molecular species and unsaturated property of FA
307
acyl chains 7. According to previous results of peach lipidome by Bustamante
308
most dramatically changed lipid belongs to galactolipid between six varieties with
309
differential chilling tolerance. Here, we found that differentially accumulated
ACS Paragon Plus Environment
20
, the
Journal of Agricultural and Food Chemistry
Page 14 of 39
310
phospholipids were enriched in the MeJA fruit with less IB (Fig 3B). Interestingly, the
311
potential biomarkers related to MeJA-induced amelioration of IB shared the same
312
lipid class, although with different fatty acyl chains 20.
313
In parallel with occurrence of cold-induced IB at the end of shelf life (Fig 1A),
314
phospholipids mainly including PCs, PEs and PGs, with linoleic acids (18:2) enriched
315
in their acyl chains, were differentially accumulated in the MeJA fruit (Fig 3B). It is
316
reported that PCs, PEs and PGs of phospholipids functioning as main components of
317
membrane systems, were associated with CI in horticultural products including peach
318
fruit
319
acyl chains in phospholipids were closely associated with cold-induced IB in peach
320
fruit 7. Previous studies regarding total FAs suggest that levels of unsaturation of FAs
321
were positively correlated with chilling tolerance in fruit including peach during cold
322
storage
323
maintained by MeJA in avocado fruit with less extent to CI during shelf life ripening
324
after cold storage
325
phospholipids may be the conserved FA adjustment by MeJA at shelf life of peach
326
fruit after cold storage.
7, 20
. In addition, our previous findings suggest that the unsaturation status of FA
13
. Recently, it is reported that higher levels of total linoleic acid (18:2) was
54
. Therefore, maintenance of levels of linoleic acid (18:2) in
327
In addition, MeJA-triggered maintenance levels of linoleic acid (18:2) in
328
phospholipids was accompanied by inhibited membrane permeability as indicated by
329
lower levels of relative electrolyte rate at the end of shelf life e (Fig 1B). Decline of
330
linoleic acid (18:2) and increase of linolenic acid (18:3) in PCs and PEs were often
331
observed concomitant with proceeding membrane leakage at the early stage of banana
332
fruit ripening and senescence 55, which were inhibited by MeJA in peach fruit
333
mentioned above. It is suggested that MeJA-triggered maintenance of levels of
ACS Paragon Plus Environment
56-57
, as
Page 15 of 39
Journal of Agricultural and Food Chemistry
334
linoleic acid (18:2) in phospholipids involve less membrane damage of CI-induced
335
ripening and senescence.
336 337
Inhibition of PpFAD8.1 but elevation of PpLOX3.1 expression by MeJA
338
contributes to unsaturation status in phospholipids and JA generation
339
The unsaturation levels of FAs are controlled by synergistic actions of FAD and 21-23
340
LOX
. The elevated accumulation of linoleic acid (18:2) in differentially
341
accumulated phospholipids coincided with lower transcript levels of PpFAD8.1 (Fig
342
4C, Fig. 5A). The similar results were observed in melting peach fruit ‘Hujingmilu’
343
with reduced IB in low temperature conditioning treatment at the end of shelf life 7.
344
PpFAD8.1 is the homologue of Arabidopsis FAD8, which is located in plastid (ω-3)
345
and can convert linoleic acid (18:2) to linolenic acid (18:3). FAD8 in plants is to be
346
considered a functional gene involved in cold stress 58-59. In the peach fruit, PpFAD8.1
347
response to cold stress was enhanced by low temperature conditioning fruit with slight
348
CI 7. Therefore, two distinct actions of PpFAD8.1 may exist in peach fruit when
349
facing different temperature environment.
350
Considering the depletion of unsaturated FAs, we further analyzed the expression
351
of PpLOXs. LOX action was closely related to JA level demonstrated by the fact that
352
linolenic acid (18:3) metabolism were significantly influenced in JA-deficient
353
Arabidopsis mutant after exogenous MeJA treatment
354
genes which can be categorized into 9-LOX and 13-LOX depending on oxygenated
355
site in the hydrocarbon backbone of FA. MeJA promoted PpLOX3.1 accumulation in
356
the transcript level, accompanied by elevated JA levels (Fig. 5B, Fig. 6A).
357 358
60
. It is encoded by multiple
PpLOX3.1 putatively belongs to 13-LOX subfamily, and act as the most abundant mRNA member in peach fruit under either cold or normal condition
ACS Paragon Plus Environment
7, 61
; it is
Journal of Agricultural and Food Chemistry
7, 57, 62
Page 16 of 39
359
responsive to cold stress and MeJA signal in peach fruit
360
tomato TomLOXE was found to be involved in JA biosynthesis to cope with abiotic
361
stress based on transgenic experiment
362
enzymatic activity of PpLOX3.1 were upregulated by exogenous MeJA in peach fruit
363
postharvest ripening
364
such as JA biosynthesis, tolerance to cold stress and fruit ripening. Moreover, there is
365
no linolenic acid (18:3) in the differentially expressed lipids (Fig. 4B-C), support the
366
notion that MeJA accelerated depletion of linolenic acid (18:3) to accumulate
367
endogenous JA via enhancing PpLOX3.1 expression in postharvest ripening after cold
368
storage.
57
63
. Its orthologue in
. Both expression of PpLOX3.1 and
. These data suggest that PpLOX3.1 may play multiple roles
369 370
MeJA promotes downstream CBF signaling pathway through PpMYC2.1
371
expression
372
The enhanced JA level was in parallel with evoked transcript levels of PpMYC2.2,
373
and PpCBF3 in MeJA fruit (Fig 6). MYC2 is a key positive effector of JA signal, and
374
CBF3 cascade has been demonstrated to be one of crucial pathways contributing to
375
cold tolerance in plants and fruit
376
MaMYC2b) physically interacted with ICE1 that can directly transactivate CBF, when
377
expression of downstream genes in CBF signaling cascades were also induced by
378
MeJA treatment 41. Recently, Wang et al. 66 reported that overexpression of MdMYC2
379
in apple calli upregulated MdCBF3 expression and enhanced freezing tolerance.
380
These data suggest that MYC2-related activation of CBF signaling pathway is a
381
general mechanism conferring cold tolerance, which may include PpMYC2.2 and
382
PpCBF3, and further enhanced by endogenous JA accumulation in peach fruit.
64-65
. In banana fruit, MYC2s (MaMYC2a and
ACS Paragon Plus Environment
Page 17 of 39
Journal of Agricultural and Food Chemistry
383
Further evidence will be needed to provide more insights into roles of PpMYC2s, and
384
associations with PpCBF3 signaling pathway.
385
In this study, details of transcript and metabolites related to phenolics metabolism
386
and lipid reprogramming were investigated to provide insights into CI amelioration by
387
MeJA in peach fruit after cold removal. Phenolics metabolism and phospholipids
388
especially PC, PE and PG were the main influenced factors by MeJA. Moreover,
389
PpFAD8.1 and PpLOX3.1 might control unsaturation status of fatty acid acyls in
390
phospholipids, and JA level. Enhanced JA further activates CBF signaling pathway
391
with participation of PpMYCs, to protect from membrane damage in peach fruit after
392
cold removal.
393 394
ABBREVIATIONS USED
395
CBF: C-repeat-binding factor; Cer: ceramide; CI: chilling injury; FAD: fatty acid
396
desaturase; GCer, glycosylceramides; IB: internal browning; JA: jasmonic acid; LOX:
397
lipoxygenase; MeJA: methyl jasmonate; PAL: phenylalanine ammonialyase; PC:
398
phosphatidylcholine; PE: phosphatidylethanolamine; PG: phosphatidylglycerol; PI:
399
phosphatidylinositol; POD: peroxidase; PPO: phenol oxidase; TG: triacylglycerol..
400 401
Funding
402
This research is supported by Anhui Provincial Department of Education Natural
403
Fund (No. KJ2018A0130), and Natural Science Foundation of Anhui province (No.
404
1808085MC94), and National Natural Science Foundation of China (No. 31772367).
405
AUTHOR INFORMATION
ACS Paragon Plus Environment
Journal of Agricultural and Food Chemistry
406
Corresponding Author
407
*Dr. Ke Wang Telephone: +86-0551-65785021 E-mail:
[email protected] 408 409
ORCID
410
https://orcid.org/0000-0003-3082-4302
411
Notes
412
All authors declare that they have no conflict of interest.
413
Supporting Information
414
Table S1. Information of primers used in real-time quantitative PCR experiment.
415
Figure S1. Relative expressions of genes through real-time quantitative PCR for
416
verification of transcriptomic data.
417
Figure S2. Content of phenolics in peach fruit stored at 0℃ for 63 days and 20℃ 6
418
days of shelf life.
419
ACKNOWLEDGMENTS
420
We thank the staff members at Omics-laboratory of the Biotechnology Center for
421
Anhui Agricultural University in Hefei, China for providing technical support in mass
422
data collection and the other valuable discussions.
423 424
REFERENCES
425
1. Aghdam, M. S.; Jannatizadeh, A.; Luo, Z.; Paliyath, G., Ensuring sufficient
426
intracellular ATP supplying and friendly extracellular ATP signaling attenuates
427
stresses, delays senescence and maintains quality in horticultural crops during
428
postharvest life. Trends Food Sci. Technol. 2018, 76, 67-81.
429
2. Genero, M.; Gismondi, M.; Monti, L. L.; Gabilondo, J.; Budde, C. O.; Andreo, C.
ACS Paragon Plus Environment
Page 18 of 39
Page 19 of 39
Journal of Agricultural and Food Chemistry
430
S.; Lara, M. V.; Drincovich, M. F.; Bustamante, C. A., Cell wall-related genes studies
431
on peach cultivars with differential susceptibility to woolliness: looking for candidates
432
as indicators of chilling tolerance. Plant Cell Rep. 2016.
433
3. Suo, J. T.; Li, H.; Ban, Q. Y.; Han, Y.; Meng, K.; Jin, M. J.; Zhang, Z. K.; Rao, J.
434
P., Characteristics of chilling injury-induced lignification in kiwifruit with different
435
sensitivities to low temperatures. Postharvest. Biol. Technol. 2018, 135, 8-18.
436
4. Ge, H.; Zhang, J.; Zhang, Y. J.; Li, X.; Yin, X. R.; Grierson, D.; Chen, K. S.,
437
EjNAC3 transcriptionally regulates chilling-induced lignification of loquat fruit via
438
physical interaction with an atypical CAD-like gene. J. Exp. Bot. 2017, 68 (18),
439
5129–5136.
440
5. Luo, Z.; Xu, X.; Yan, B., Use of 1-methylcyclopropene for alleviating chilling
441
injury and lignification of bamboo shoot (Phyllostachys praecox f. prevernalis) during
442
cold storage. J. Sci. Food Agric. 2008, 88 (1), 151-157.
443
6. Karagiannis, E.; Michailidis, M.; Tanou, G.; Samiotaki, M.; Karamanoli, K.;
444
Avramidou, E.; Ganopoulos, I.; Madesis, P.; Molassiotis, A., Ethylene –dependent and
445
–independent superficial scald resistance mechanisms in ‘Granny Smith’ apple fruit.
446
Sci. Rep. 2018, 8 (1), 11436.
447
7. Wang, K.; Yin, X. R.; Zhang, B.; Grierson, D.; Xu, C. J.; Chen, K. S.,
448
Transcriptomic and metabolic analyses provide new insights into chilling injury in
449
peach fruit. Plant Cell Environ. 2017, 40 (8), 1531-1551.
450
8. Luengwilai, K.; Beckles, D. M.; Roessner, U.; Dias, D. A.; Lui, V.; Siriphanich, J.,
451
Identification of physiological changes and key metabolites coincident with
ACS Paragon Plus Environment
Journal of Agricultural and Food Chemistry
452
postharvest internal browning of pineapple (Ananas comosus L.) fruit. Postharvest.
453
Biol. Technol. 2018, 137, 56-65.
454
9. Deuchande, T.; Larrigaudiere, C.; Gine-Bordonaba, J.; Carvalho, S. M. P.;
455
Vasconcelos, M. W., Biochemical basis of CO2-related internal browning disorders in
456
pears (pyrus communis l. Cv. Rocha) during long-term storage. J. Agric. Food Chem.
457
2016, 64 (21), 4336-4345.
458
10. Luo, Z.; Li, D.; Du, R.; Mou, W., Hydrogen sulfide alleviates chilling injury of
459
banana fruit by enhanced antioxidant system and proline content. Sci Hortic 2015,
460
183, 144-151.
461
11. Sevillano, L.; Sanchez-Ballesta, M. T.; Romojaro, F.; Flores, F. B., Physiological,
462
hormonal and molecular mechanisms regulating chilling injury in horticultural species.
463
Postharvest technologies applied to reduce its impact. J. Sci. Food Agric. 2009, 89 (4),
464
555-573.
465
12. Toivonen, P. M. A.; Brummell, D. A., Biochemical bases of appearance and
466
texture changes in fresh-cut fruit and vegetables. Postharvest. Biol. Technol. 2008, 48
467
(1), 1-14.
468
13. Zhang, C. F.; Tian, S. P., Crucial contribution of membrane lipids' unsaturation to
469
acquisition of chilling-tolerance in peach fruit stored at 0°C. Food Chem. 2009, 115
470
(2), 405-411.
471
14. Zhang, C. F.; Tian, S. P., Peach fruit acquired tolerance to low temperature stress
472
by accumulation of linolenic acid and N-acylphosphatidylethanolamine in plasma
473
membrane. Food Chem. 2010, 120 (3), 864-872.
ACS Paragon Plus Environment
Page 20 of 39
Page 21 of 39
Journal of Agricultural and Food Chemistry
474
15. Lin, Y.; Lin, Y.; Lin, H.; Shi, J.; Chen, Y.; Wang, H., Inhibitory effects of propyl
475
gallate on membrane lipids metabolism and its relation to increasing storability of
476
harvested longan fruit. Food Chem. 2017, 217, 133-138.
477
16. Lafuente, M. T.; Estables-Ortiz, B.; Gonzalez-Candelas, L., Insights into the
478
molecular events that regulate heat-induced chilling tolerance in citrus fruits.
479
Frontiers in Plant Science 2017, 8.
480
17. Zheng, G.; Li, L.; Li, W., Glycerolipidome responses to freezing- and
481
chilling-induced injuries: examples in Arabidopsis and rice. BMC Plant Biol. 2016, 16
482
(1), 70.
483
18. Kong, X. M.; Wei, B. D.; Gao, Z.; Zhou, Y.; Shi, F.; Zhou, X.; Zhou, Q.; Ji, S. J.,
484
Changes in membrane lipid composition and function accompanying chilling injury in
485
bell peppers. Plant Cell Physiol. 2018, 59 (1), 167-178.
486
19. Marla, S. R.; Shiva, S.; Welti, R.; Liu, S. Z.; Burke, J. J.; Morris, G. P.,
487
Comparative transcriptome and lipidome analyses reveal molecular chilling responses
488
in chilling-tolerant Sorghums. Plant Genome-Us 2017, 10 (3).
489
20. Bustamante, C. A.; Brotman, Y.; Monti, L. L.; Gabilondo, J.; Budde, C. O.; Lara,
490
M. V.; Fernie, A. R.; Drincovich, M. F., Differential lipidome remodeling during
491
postharvest of peach varieties with different susceptibility to chilling injury. Physiol.
492
Plant. 2018, 163 (1), 2-17.
493
21. Aghdam, M. S.; Bodbodak, S., Physiological and biochemical mechanisms
494
regulating chilling tolerance in fruits and vegetables under postharvest salicylates and
495
jasmonates treatments. Sci Hortic 2013, 156, 73-85.
ACS Paragon Plus Environment
Journal of Agricultural and Food Chemistry
Page 22 of 39
496
22. Jannatizadeh, A.; Aghdam, M. S.; Luo, Z.; Razavi, F., Impact of Exogenous
497
Melatonin Application on Chilling Injury in Tomato Fruits During Cold Storage. Food
498
Bioprocess Tech 2019, 12 (5), 741-750.
499
23. Jiao,
500
Nitric-Oxide-Induced Chilling Tolerance and Defense Response in Postharvest Peach
501
Fruit. J. Agric. Food Chem. 2019, 67 (17), 4764-4773.
502
24. Alhassan, N.; Golding, J. B.; Wills, R. B. H.; Bowyer, M. C.; Pristijono, P., Long
503
Term Exposure to Low Ethylene and Storage Temperatures Delays Calyx Senescence
504
and Maintains ‘Afourer’ Mandarins and Navel Oranges Quality. Foods 2019, 8 (1),
505
19.
506
25. Zhang, Q. T.; Zhang, L. L.; Geng, B.; Feng, J. R.; Zhu, S. H., Interactive effects
507
of abscisic acid and nitric oxide on chilling resistance and active oxygen metabolism
508
in peach fruit during cold storage. J. Sci. Food Agric. 2019, 99 (7), 3367-3380.
509
26. Ding, Y.; Zhao, J. H.; Nie, Y.; Fan, B.; Wu, S. J.; Zhang, Y.; Sheng, J. P.; Shen, L.;
510
Zhao, R. R.; Tang, X. M., Salicylic-Acid-Induced Chilling- and Oxidative-Stress
511
Tolerance in Relation to Gibberellin Homeostasis, C-Repeat/Dehydration-Responsive
512
Element Binding Factor Pathway, and Antioxidant Enzyme Systems in Cold-Stored
513
Tomato Fruit. J. Agric. Food Chem. 2016, 64 (43), 8200-8206.
514
27. Zhu, Z.; Ding, Y.; Zhao, J.; Nie, Y.; Zhang, Y.; Sheng, J.; Tang, X., Effects of
515
Postharvest Gibberellic Acid Treatment on Chilling Tolerance in Cold-Stored Tomato
516
(Solanum lycopersicum L.) Fruit. Food Bioprocess Tech 2016, 9 (7), 1202-1209.
517
28. Gao, H.; Lu, Z. M.; Yang, Y.; Wang, D. N.; Yang, T.; Cao, M. M.; Cao, W.,
C.;
Chai,
Y.;
Duan,
Y.,
Inositol
1,4,5-Trisphosphate
ACS Paragon Plus Environment
Mediates
Page 23 of 39
Journal of Agricultural and Food Chemistry
518
Melatonin treatment reduces chilling injury in peach fruit through its regulation of
519
membrane fatty acid contents and phenolic metabolism. Food Chem. 2018, 245,
520
659-666.
521
29. Gao, H.; Zhang, Z. K.; Lv, X. G.; Cheng, N.; Peng, B. Z.; Cao, W., Effect of
522
24-epibrassinolide on chilling injury of peach fruit in relation to phenolic and proline
523
metabolisms. Postharvest. Biol. Technol. 2016, 111, 390-397.
524
30. Liu, Z.; Li, L.; Luo, Z.; Zeng, F.; Jiang, L.; Tang, K., Effect of brassinolide on
525
energy status and proline metabolism in postharvest bamboo shoot during chilling
526
stress. Postharvest. Biol. Technol. 2016, 111, 240-246.
527
31. Aghdam, M. S.; Asghari, M.; Khorsandi, O.; Mohayeji, M., Alleviation of
528
postharvest chilling injury of tomato fruit by salicylic acid treatment. Journal of food
529
science and technology 2014, 51 (10), 2815-20.
530
32. Sayyari, M.; Babalar, M.; Kalantari, S.; Serrano, M.; Valero, D., Effect of
531
salicylic acid treatment on reducing chilling injury in stored pomegranates.
532
Postharvest. Biol. Technol. 2009, 53 (3), 152-154.
533
33. Luo, Z. S.; Chen, C.; Xie, J., Effect of salicylic acid treatment on alleviating
534
postharvest chilling injury of 'Qingnai' plum fruit. Postharvest. Biol. Technol. 2011,
535
62 (2), 115-120.
536
34. Cao, S. F.; Zheng, Y. H.; Wang, K. T.; Rui, H. J.; Tang, S. S., Effect of methyl
537
jasmonate on cell wall modification of loquat fruit in relation to chilling injury after
538
harvest. Food Chem. 2010, 118 (3), 641-647.
539
35. Cai, Y. T.; Cao, S. F.; Yang, Z. F.; Zheng, Y. H., MeJA regulates enzymes
ACS Paragon Plus Environment
Journal of Agricultural and Food Chemistry
540
involved in ascorbic acid and glutathione metabolism and improves chilling tolerance
541
in loquat fruit. Postharvest. Biol. Technol. 2011, 59 (3), 324-326.
542
36. Rehman, M.; Singh, Z.; Khurshid, T., Methyl jasmonate alleviates chilling injury
543
and regulates fruit quality in 'Midknight' Valencia orange. Postharvest. Biol. Technol.
544
2018, 141, 58-62.
545
37. Sayyari, M.; Babalar, M.; Kalantari, S.; Martinez-Romero, D.; Guillen, F.;
546
Serrano, M.; Valero, D., Vapour treatments with methyl salicylate or methyl
547
jasmonate alleviated chilling injury and enhanced antioxidant potential during
548
postharvest storage of pomegranates. Food Chem. 2011, 124 (3), 964-970.
549
38. Zhang, X. H.; Sheng, J. P.; Li, F. J.; Meng, D. M.; Shen, L., Methyl jasmonate
550
alters arginine catabolism and improves postharvest chilling tolerance in cherry
551
tomato fruit. Postharvest. Biol. Technol. 2012, 64 (1), 160-167.
552
39. Min, D.; Li, F.; Zhang, X.; Cui, X.; Shu, P.; Dong, L.; Ren, C., SlMYC2 Involved
553
in Methyl Jasmonate-Induced Tomato Fruit Chilling Tolerance. J. Agric. Food Chem.
554
2018, 66 (12), 3110-3117.
555
40. Rivera-Dominguez, M.; Astorga-Cienfuegos, K. R.; Tiznado-Hernandez, M. E.;
556
Gonzalez-Aguilar, G. A., Induction of the expression of defence genes in Carica
557
papaya fruit by methyl jasmonate and low temperature treatments. Electron. J.
558
Biotechnol. 2012, 15 (5).
559
41. Zhao, M. L.; Wang, J. N.; Shan, W.; Fan, J. G.; Kuang, J. F.; Wu, K. Q.; Li, X. P.;
560
Chen, W. X.; He, F. Y.; Chen, J. Y.; Lu, W. J., Induction of jasmonate signalling
561
regulators MaMYC2s and their physical interactions with MaICE1 in methyl
ACS Paragon Plus Environment
Page 24 of 39
Page 25 of 39
Journal of Agricultural and Food Chemistry
562
jasmonate-induced chilling tolerance in banana fruit. Plant Cell Environ. 2013, 36 (1),
563
30-51.
564
42. Li, H.; Suo, J. T.; Han, Y.; Liang, C. Q.; Jin, M. J.; Zhang, Z. K.; Rao, J. P., The
565
effect of 1-methylcyclopropene, methyl jasmonate and methyl salicylate on lignin
566
accumulation and gene expression in postharvest 'Xuxiang' kiwifruit during cold
567
storage. Postharvest. Biol. Technol. 2017, 124, 107-118.
568
43. Jin, P.; Zhu, H.; Wang, J.; Chen, J. J.; Wang, X. L.; Zheng, Y. H., Effect of methyl
569
jasmonate on energy metabolism in peach fruit during chilling stress. J. Sci. Food
570
Agric. 2013, 93 (8), 1827-1832.
571
44. Yu, L. N.; Liu, H. X.; Shao, X. F.; Yu, F.; Wei, Y. Z.; Ni, Z. M.; Xu, F.; Wang, H.
572
F., Effects of hot air and methyl jasmonate treatment on the metabolism of soluble
573
sugars in peach fruit during cold storage. Postharvest. Biol. Technol. 2016, 113, 8-16.
574
45. Meng, X. H.; Han, J.; Wang, Q.; Tian, S. P., Changes in physiology and quality of
575
peach fruits treated by methyl jasmonate under low temperature stress. Food Chem.
576
2009, 114 (3), 1028-1035.
577
46. Ding, C. K.; Wang, C. Y.; Gross, K. C.; Smith, D. L., Reduction of chilling injury
578
and transcript accumulation of heat shock proteins in tomato fruit by methyl
579
jasmonate and methyl salicylate. Plant Sci. 2001, 161 (6), 1153-1159.
580
47. Jin, P.; Wang, K. U.; Shang, H. T.; Tong, J. M.; Zheng, Y. H., Low-temperature
581
conditioning combined with methyl jasmonate treatment reduces chilling injury of
582
peach fruit. J. Sci. Food Agric. 2009, 89 (10), 1690-1696.
583
48. Kondo, S.; Kittikorn, M.; Kanlayanarat, S., Preharvest antioxidant activities of
ACS Paragon Plus Environment
Journal of Agricultural and Food Chemistry
584
tropical fruit and the effect of low temperature storage on antioxidants and jasmonates.
585
Postharvest. Biol. Technol. 2005, 36 (3), 309-318.
586
49. Wang, K.; Shao, X. F.; Gong, Y. F.; Zhu, Y.; Wang, H. F.; Zhang, X. L.; Yu, D. D.;
587
Yu, F.; Qiu, Z. Y.; Lu, H., The metabolism of soluble carbohydrates related to chilling
588
injury in peach fruit exposed to cold stress. Postharvest. Biol. Technol. 2013, 86,
589
53-61.
590
50. Peng, J.; Zhu, H.; Wang, J.; Chen, J.; Wang, X.; Zheng, Y., Effect of methyl
591
jasmonate on energy metabolism in peach fruit during chilling stress. J. Sci. Food
592
Agric. 2013, 93 (8), 1827-1832.
593
51. Peng, J.; Zheng, Y. H.; Tang, S. S.; Rui, H. J.; Wang, C. Y., A combination of hot
594
air and methyl jasmonate vapor treatment alleviates chilling injury of peach fruit.
595
Postharvest. Biol. Technol. 2009, 52 (1), 24–29.
596
52. Jin, P.; Zheng, Y. H.; Tang, S. S.; Rui, H. J.; Wang, C. Y., A combination of hot air
597
and methyl jasmonate vapor treatment alleviates chilling injury of peach fruit.
598
Postharvest. Biol. Technol. 2009, 52 (1), 24-29.
599
53. Vicente, A. R.; Pineda, C.; Lemoine, L.; Civello, P. M.; Martinez, G. A.; Chaves,
600
A. R., UV-C treatments reduce decay, retain quality and alleviate chilling injury in
601
pepper. Postharvest. Biol. Technol. 2005, 35 (1), 69-78.
602
54. Glowacz, M.; Bill, M.; Tinyane, P. P.; Sivakumar, D., Maintaining postharvest
603
quality of cold stored 'Hass' avocados by altering the fatty acids content and
604
composition with the use of natural volatile compounds - methyl jasmonate and
605
methyl salicylate. J. Sci. Food Agric. 2017, 97 (15), 5186-5193.
ACS Paragon Plus Environment
Page 26 of 39
Page 27 of 39
Journal of Agricultural and Food Chemistry
606
55. Wade, N. L.; Bishop, D. G., Changes in the lipid composition of ripening banana
607
fruits and evidence for an associated increase in cell membrane permeability. Biochim.
608
Biophys. Acta 1978, 529 (3), 454-460.
609
56. Soto, A.; Ruiz, K. B.; Ziosi, V.; Costa, G.; Torrigiani, P., Ethylene and auxin
610
biosynthesis and signaling are impaired by methyl jasmonate leading to a transient
611
slowing down of ripening in peach fruit. J. Plant Physiol. 2012, 169 (18), 1858-65.
612
57. Wei, J. X.; Wen, X. C.; Tang, L., Effect of methyl jasmonic acid on peach fruit
613
ripening progress. Sci Hortic 2017, 220, 206-213.
614
58. Soria-Garci, A. I.; Rubio, M. A. C.; Lagunas, B.; Li Pez-Gomolli, N. S.; Luji, N.
615
M.; Di Az-Guerra, R. L.; Picorel, R.; Alfonso, M., Tissue Distribution and Specific
616
Contribution of Arabidopsis FAD7 and FAD8 Plastid Desaturases to the JA- and
617
ABA-Mediated Cold Stress or Defense Responses. Plant Cell Physiol. 2019, 60 (5),
618
1025-1040.
619
59. Liu, W.; Li, W.; He, Q. L.; Daud, M. K.; Chen, J. H.; Zhu, S. J., Characterization
620
of 19 genes encoding membrane-bound fatty acid desaturases and their expression
621
profiles in gossypium raimondii under low temperature. PLoS One 2015, 10 (4).
622
60. Cao, J. J.; Li, M. Y.; Chen, J.; Liu, P.; Li, Z., Effects of MeJA on Arabidopsis
623
metabolome under endogenous JA deficiency. Sci. Rep. 2016, 6, 37674.
624
61. Guo, S. L.; Song, Z. Z.; Ma, R. J.; Yang, Y.; Yu, M. L., Genome-wide
625
identification and expression analysis of the lipoxygenase gene family during peach
626
fruit ripening under different postharvest treatments. Acta Physiol Plant 2017, 39 (5).
627
62. Han, M. Y.; Zhang, T.; Zhao, C. P.; Zhi, J. H., Regulation of the expression of
ACS Paragon Plus Environment
Journal of Agricultural and Food Chemistry
628
lipoxygenase genes in Prunus persica fruit ripening. Acta Physiol Plant 2011, 33 (4),
629
1345-1352.
630
63. Hu, T. Z.; Hu, Z. L.; Zeng, H.; Qv, X. X.; Chen, G. P., Tomato lipoxygenase D
631
involved in the biosynthesis of jasmonic acid and tolerance to abiotic and biotic stress
632
in tomato. Plant Biotechnol Rep 2015, 9 (1), 37-45.
633
64. Gilmour, S. J.; Sebolt, A. M.; Salazar, M. P.; Everard, J. D.; Thomashow, M. F.,
634
Overexpression of the Arabidopsis CBF3 transcriptional activator mimics multiple
635
biochemical changes associated with cold acclimation. Plant Physiol. 2000, 124 (4),
636
1854-65.
637
65. Zhang, T.; Che, F. B.; Zhang, H.; Pan, Y.; Xu, M. Q.; Ban, Q. Y.; Han, Y.; Rao, J.
638
P., Effect of nitric oxide treatment on chilling injury, antioxidant enzymes and
639
expression of the CmCBF1 and CmCBF3 genes in cold-stored Hami melon (Cucumis
640
melo L.) fruit. Postharvest. Biol. Technol. 2017, 127, 88-98.
641
66. Wang, Y.; Xu, H.; Liu, W.; Wang, N.; Qu, C.; Jiang, S.; Fang, H.; Zhang, Z.; Chen,
642
X., Methyl jasmonate enhances apple’ cold tolerance through the JAZ–MYC2
643
pathway. Plant Cell, Tissue and Organ Culture (PCTOC) 2019, 136 (1), 75-84.
644 645
ACS Paragon Plus Environment
Page 28 of 39
Page 29 of 39
Journal of Agricultural and Food Chemistry
646
Figure Captions
647
Fig 1. IB index (A) and electrolyte leakage rate (B) in peach fruit stored at 0℃ for 63
648
days and 6 days shelf life. Asterisks denote significant levels in the comparison
649
between means of MeJA and control (*, p