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作物学报

作物学报 ›› 2019, Vol. 45 ›› Issue (2): 235-247.doi: 10.3724/SP.J.1006.2019.84063

• 耕作栽培·生理生化 • 上一篇    下一篇

挥发性抑芽物质对马铃薯块茎萌芽的影响及其作用机制

邹雪1,2,丁凡1,余金龙1,彭洁2,邓孟胜2,王宇2,刘丽芳1,余韩开宗1,陈年伟1,王西瑶2,*()   

  1. 1绵阳市农业科学研究院, 四川绵阳 621023
    2四川农业大学农学院, 四川成都 611130
  • 收稿日期:2018-04-29 接受日期:2018-08-20 出版日期:2019-02-12 网络出版日期:2018-09-26
  • 通讯作者: 王西瑶
  • 基金资助:
    本研究由四川省科技厅公益性育种攻关项目(2016NYZ0032);绵阳市农业科学研究院创新基金项目(cxjj462016-2019)

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 Published:2019-02-12 Published online:2018-09-26
  • Contact: Xi-Yao WANG
  • 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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摘要：

马铃薯块茎过早萌芽会降低商品价值, 本试验比较挥发性物质的抑芽效应, 并从转录和蛋白水平分析其作用机制。结果表明, 抑芽能力薄荷醇?樟脑?萘, 萌芽受抑降低代谢消耗, 薄荷醇处理180 d时的重量损失只有对照损失的36%。樟脑处理萌芽块茎3 d引起表达显著上调(或下调)的基因和蛋白分别有1227 (299)和296 (204)个, 主要参与响应刺激、防御反应。贮藏期间, 果胶代谢基因PEL、PME、PG, 角质合成基因CYP77A6、HPFT、WES, 乙烯合成基因ACO以及转录因子编码基因GATA4L的表达量随时间升高。樟脑早期不同程度地刺激这些基因表达, 中后期则抑制, 49 d时只有对照的0.68%~23.35%。薄荷醇使上述基因表达保持低水平, 但可提高细胞周期负控基因KRP4的表达, 为对照的15.9倍。植物病原菌互作通路中的基因WRKY75、STH-2、RBOH表达受樟脑诱导并在后期高表达。樟脑和薄荷醇均能抑制生长发育基因的表达, 造成芽死亡, 降低贮藏损耗。前者早期能促进合成保护性物质, 萌芽时有更强烈的抗菌反应; 后者则阻碍细胞分裂。

关键词: 马铃薯, 贮藏, 抑芽物质, 转录组, 蛋白组

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

图1

用作转录组和蛋白组测序的处理方式"

表1

试验选择基因的引物序列"

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

图2

贮藏50 d时各处理的抑芽作用比较 CK: 对照; NAP: 萘处理; CAM: 樟脑处理; MEN: 薄荷醇处理。"

表2

贮藏期间各处理的平均芽长及块茎重量损失"

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

图3

荧光定量验证转录组测序"

图4

樟脑处理的差异表达基因和蛋白的显著GO富集"

表3

RNA-seq测序得到的差异基因富集到的前10个代谢通路"

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 p