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Acta Agronomica Sinica ›› 2019, Vol. 45 ›› Issue (2): 235-247.doi: 10.3724/SP.J.1006.2019.84063

• TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY • Previous Articles     Next Articles

Suppression mechanism of volatile sprout-inhibitors on potato tuber sprouting

Xue ZOU1,2,Fan DING1,Jin-Long YU1,Jie PENG2,Meng-Sheng DENG2,Yu WANG2,Li-Fang LIU1,Kai-Zong YU-HAN1,Nian-Wei CHEN1,Xi-Yao WANG2,*()   

  1. 1 Mianyang Academy of Agricultural Sciences, Mianyang 621023, Sichuan, China
    2 College of Agronomy, Sichuan Agricultural University, Chengdu 611130, Sichuan, China
  • Received:2018-04-29 Accepted:2018-08-20 Online:2019-02-12 Published:2018-09-26
  • Contact: Xi-Yao WANG E-mail:wxyrtl@163.com
  • Supported by:
    This study was supported by the Breeding Program for Public Welfare of Science & Technology Department of Sichuan Province(2016NYZ0032);and the Innovation Fund of Mianyang Academy of Agricultural Sciences(cxjj462016-2019)

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Abstract:

Tuber sprouting losses commercial value. In this experiment, the sprout inhibition abilities of naphthalene, camphor, menthol were researched and the acting mechanism by RNA-seq, iTRAQ was explained. The inhibition abilities showed a trend of menthol > camphor > naphthalene. The metabolic consumption was reduced due to sprouting inhibition and the tuber weight loss of menthol treatment was only 36% of the control loss in 180 days storage. Compared with the control, about 1227 (299) and 296 (204) genes and proteins whose expression levels were significantly up-regulated (or down-regulated) were detected respectively in sprouting tuber with camphor treatment for three days. Those genes and proteins mainly involved in response to stimulus and defense response. Transcripts of PEL, PME, PG related to pectin degradation, CYP77A6, HPFT, WES related to cutin synthesis, ACO related to synthesis of ethylene, and GATA4L coding transcription factor were upward with dormancy release during storage. Relative expression of those genes was stimulated at different degrees in camphor treatment at the early stage, but significantly inhibited at the middle and late stages, showing 0.68%-23.35% of the control expression at 49 days. Menthol treatment maintained these genes with low level expression in tuber, but significantly increased the expression of cell cycle inhibitor gene KRP4 to 15.9 times of control. Camphor treatment increased transcripts of genes WRKY75, STH-2, RBOH participating in plant-pathogen interaction pathway to the highest levels at sprouting stage. Therefore, we can conclude that camphor and menthol suppress the growth and development of sprouting tuber, eventually kill the bud and reduce the tuber weight loss during storage. Camphor treatment promotes the biosynthesis of protective substances to resist stress at the early stage and strengthen the resistance to pathogen infection at the sprouting stage; menthol treatment prevents bud sprouting by increasing KRP4 expression to inhibit cell division.

Key words: potato, storage, sprout-inhibitors, transcriptome, proteome

Fig. 1

Treatment schematics for the RNA-seq and iTRAQ experimental samples are listed"

Table 1

Primer sequences used in this study"

基因
Gene		 编号
ID		 引物
Primers (5°-3°)		 长度
Length
(bp)		 熔解温度
Melt temp.
(℃)
延伸因子EF1αL (内参Reference gene)
Elongation factor 1 alpha-like		 PGSC0003DMT400014674 CTTGTACACCACGCTAAGGAG
GTCAATGCAAACCATTCCTTG		 155 81.5
果胶裂解酶 PEL
Pectate lyase		 PGSC0003DMT400051938 TGGGAAGGATCATGGAAACA
ACCAGCATTTGAGGTAATCATT		 168 83.0
果胶酯酶PME
Pectinesterase		 PGSC0003DMT400005343 AATCTACTGGTGTCGCTTAC
TATTGTGGCATTGGTTCA		 167 78.5
多聚半乳糖醛酸酶PG
Polygalacturonase		 PGSC0003DMT400035162 CAAGTAAAGGCTACGACAA
TATGGACCTGAGAATGAAA		 199 81.0
细胞色素P450, CYP77A6
Cytochrome P450, CYP77A6		 PGSC0003DMT400060106 CTTAGTCCACGAAGCATT
TTCCGTAGTGACCTCCA		 141 84.0
ω-羟基酸O-阿魏酸酯转移酶HPFT
Omega-hydroxypalmitate O-feruloyl
transferase		 PGSC0003DMT400020180 AGGTCACTTATCCGTTCC
GAGTTGGCAATTTCAGG		 110 79.0
甘油二酯O-酰基转移酶WES
Diacylglycerol O-acyltransferase		 PGSC0003DMT400023554 GTGTTGACCCGATTACCCA
CTTGTTTAGTGACAGCCTCTT		 295 81.5
AG-基序结合蛋白 GATA4L
GATA transcription factor 4-like		 PGSC0003DMT400030708 GGTTGTTACTACGATGCTCT
AATTATTGGATGTATCTTCCAC		 121 80.5
Kip相关蛋白4 KRP4
Kip-related protein 4		 PGSC0003DMT400019919 AAAAGACAGAATCGGAGTT
GTCCTCTCTCTACCATCAAT		 182 81.0
氨基环丙烷羧酸氧化酶ACO
Aminocyclopropanecarboxylate oxidase		 PGSC0003DMT400030676 CAGAGGAGCATCGTTACTGG
AAAGAACCCTGCCTCCAAAC		 194 82.0
转录因子WRKY 75
WRKY transcription factor 75		 PGSC0003DMT400056352 CAGCATCATCGTCGTCAT
CATTAGTCCCAAGAACCC		 131 77.5
病程相关蛋白STH-2
Pathogenesis-related protein STH-2		 PGSC0003DMT400003934 AAAACCAGGCATGGAACT
ACGTGTAGACCTGATTCTTT		 99 79.0
呼吸爆发氧化酶(NADPH氧化酶) RBOH
Respiratory burst oxidase (NADPH oxidase)		 PGSC0003DMT400066027 CTCTCATCACCATGCTCCAGG
TGGCAGTGTCGTCTACCATAT		 168 82.5

Fig. 2

Sprouting inhibition comparison of different treatments at 50 days CK: control; NAP: naphthalene treatment; CAM: camphor treatment; MEN: menthol treatment."

Table 2

Shoot length and tuber weight loss of different treatments during storage"

处理
Treatment		 芽长 Shoot length (mm) 块茎重量损失 Tuber weight loss (%)
50 d 65 d 80 d 60 d 120 d 180 d
CK 2.45±0.36 aA 6.02±0.37 A 14.29±1.33 A 6.68±0.36 aA 13.85±0.61 A 22.78±0.99 A
NAP 2.28±0.36 abA 2.23±0.27 B 2.06±0.47 B 6.97±0.58 aA 11.88±0.92 B 17.49±0.93 B
CAM 1.59±0.45 bA 1.31±0.20 C 凋亡Death 5.67±0.36 bAB 9.42±0.67 C 14.30±1.17 C
MEN 无芽生长 No bud growth 4.83±0.27 cB 7.37±0.55 D 8.14±1.01 D

Fig. 3

Quantitative RT-PCR validation of RNA-seq"

Fig. 4

Significant GO terms analysis of differentially expressed genes and proteins in camphor treatment"

Table 3

The first 10 pathways of differentially expressed genes in RNA-seq"

代谢路径
Pathway		 注释到该路径的差异基因数
DEGs with pathway annotation		 P值
P-value		 路径ID号
Pathway ID
谷胱甘肽代谢Glutathione metabolism 23↑ 1↓ 5.32E-08 ko00480
异黄酮生物合成Isoflavonoid biosynthesis 15↑ 0↓ 4.61E-05 ko00943
玉米素生物合成Zeatin biosynthesis 26↑ 2↓ 5.88E-05 ko00908
二苯乙烯类化合物、二芳基庚烷和姜辣素生物合成
Stilbenoid, diarylheptanoid and gingerol biosynthesis		 25↑ 10↓ 9.78E-05 ko00945
柠檬烯和蒎烯降解Limonene and pinene degradation 22↑ 5↓ 3.16E-04 ko00903
植物病原菌互作Plant-pathogen interaction 101↑ 8↓ 3.21E-04 ko04626
内质网蛋白质加工 Protein processing in endoplasmic reticulum 25↑ 15↓ 3.97E-04 ko04141
黄酮和黄酮醇生物合成Flavone and flavonol biosynthesis 21↑ 1↓ 9.68E-04 ko00944
苯丙素生物合成Phenylpropanoid biosynthesis 27↑ 10↓ 2.75E-03 ko00940
次生代谢物的生物合成 Biosynthesis of secondary metabolites 117↑ 37↓ 5.05E-03 ko01110

Fig. 5

Correlation analyses in GO enrichment of genes with same expression trend between transcriptional and proteomics groups"

Fig. 6

Effects of sprouting-inhibitors on the expression of genes related to pectin metabolism and cutin, suberine, wax biosynthesis CK: control; CAM: camphor treatment; MEN: menthol treatment; CK-4℃: control tuber storage at 4℃; the error line was the standard deviation of three repeated values."

Fig. 7

Effects of sprouting-inhibitors on the expression of genes related to growth The abbreviations of different treatments are the same as those given in Fig. 6; the error line was the standard deviation of three repeated values."

Fig. 8

Effects of sprouting-inhibitors on the expression of genes related to plant-pathogen interaction The abbreviations of different treatments are the same as those given in Fig. 6; the error line was the standard deviation of three repeated values."

Fig. 9

Acting pattern speculation of tuber response to camphor treatment Solid arrows: expression levels up-regulated compared with control; dotted arrows: expression levels down-regulated compared with control, the size of the arrow represented the degree of variability."

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