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浙江大学学报（农业与生命科学版）
浙江大学学报（农业与生命科学版）  2019, Vol. 45 Issue (4): 418-425    DOI: 10.3785/j.issn.1008-9209.2018.10.151
植物保护     
草莓响应炭疽菌侵染的差异磷酸化蛋白质组学分析
余红1(),严建立1,裘劼人1,王淑珍1,忻雅1,童建新1,来文国1,方献平1,2()
1. 杭州市农业科学研究院生物技术研究所，杭州 310024
2. 上海市农业科学院林木果树研究所，上海 201403
Differential phosphoproteomic analysis of strawberry in response to Colletotrichum gloeosporioides
Hong YU1(),Jianli YAN1,Jieren QIU1,Shuzhen WANG1,Ya XIN1,Jianxin TONG1,Wenguo LAI1,Xianping FANG1,2()
1. Institute of Biotechnology, Hangzhou Academy of Agricultural Sciences, Hangzhou 310024, China
2. Institute of Forestry and Pomology, Shanghai Academy of Agricultural Sciences, Shanghai 201403, China
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摘要：

以不同炭疽病抗性的草莓品种‘红颊’和‘甜查理’的茎组织为试验材料，采用非标记定量磷酸化蛋白质组学技术分析其在炭疽菌侵染胁迫后磷酸化蛋白组的变化。结果表明，‘红颊’和‘甜查理’中分别有154个和173个磷酸化蛋白在病菌侵染后发生了1.5倍以上的差异表达。功能注释归类分析发现，大部分差异磷酸化蛋白参与了大分子复合体形成及结构定位、刺激应答和信号转导等生物学过程。生物信息学分析进一步表明，相对于易感品种‘红颊’，高抗品种‘甜查理’差异磷酸化蛋白特异性表现出S*Y和T*F 2种保守结构域类型，且植物激素信号传导途径和碳固定途径都发生了更高程度的磷酸化。本研究结果揭示出，这些信号通路在防御病菌入侵过程中发挥了重要作用，为后续深入揭示草莓-炭疽病菌互作分子机制及草莓抗病新品种选育奠定了宝贵的理论研究基础。
关键词： 草莓;  胶孢炭疽菌;  磷酸化蛋白质组学;  ‘红颊’;  ‘甜查理’    
Abstract:

Label-free based phosphoproteomics technology was used to study the phosphoproteome change of disease-susceptible strawberry cultivar ‘Benihopp’ and resistant cultivar ‘Sweet Charlie’ infected with Colletotrichum gloeosporioides. There were 154 and 173 phosphoproteins showing more than 1.5-fold change in ‘Benihopp’ and ‘Sweet Charlie’, respectively. The differentially expressed phosphoproteins were analyzed by gene ontology annotation and bioinformatics, and we found that, most differentially expressed phosphoproteins were involved in macromolecule complexe formation, and the biological processes of structural localization, stimulus response and signal transduction. Compared with ‘Benihopp’, ‘Sweet Charlie’ may specifically own two motif types of S*Y and T*F of differentially expressed phosphorylated peptides, and its plant hormone signal transduction pathway and carbon fixation pathway had higher levels of phosphorylation. The result is of great benefit to further deeply reveal the molecular mechanisms of plant-pathogen interactions and the breeding of pathogen-resistant strawberry cultivars.
Key words: strawberry    Colletotrichum gloeosporioides    phosphoproteomics    ‘Benihopp’    ‘Sweet Charlie’
收稿日期: 2018-10-15 出版日期: 2019-09-17
CLC:  S 668.4  
基金资助: 浙江省农业（果品）新品种选育重大科技专项“草莓新品种选育”(2016C02052-8);浙江省特色农产品全产业链安全风险管控项目(HZHNKY2018001);杭州市重大科技项目(20162012A2);杭州市科技发展计划项目(20170432B14)
通讯作者: 方献平     E-mail: 340292062@qq.com;fxpbio@163.com
作者简介: 余红（https://orcid.org/0000-0001-6255-6635），E-mail：340292062@qq.com
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余红
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童建新
来文国
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引用本文:

余红, 严建立, 裘劼人, 王淑珍, 忻雅, 童建新, 来文国, 方献平. 草莓响应炭疽菌侵染的差异磷酸化蛋白质组学分析[J]. 浙江大学学报（农业与生命科学版）, 2019, 45(4): 418-425.

Hong YU, Jianli YAN, Jieren QIU, Shuzhen WANG, Ya XIN, Jianxin TONG, Wenguo LAI, Xianping FANG. Differential phosphoproteomic analysis of strawberry in response to Colletotrichum gloeosporioides. Journal of Zhejiang University (Agriculture and Life Sciences), 2019, 45(4): 418-425.

链接本文:

http://www.zjujournals.com/agr/CN/10.3785/j.issn.1008-9209.2018.10.151    或    http://www.zjujournals.com/agr/CN/Y2019/V45/I4/418

图1  病菌易感草莓品种‘红颊’和抗性品种‘甜查理’在对照和接种炭疽病菌后的病征变化
图2  鉴定到的磷酸化位点分布统计图

草莓品种

Strawberry cultivar

差异倍数

Fold change

上调肽段数

Number of up-regulated peptides

下调肽段数

Number of down-regulated peptides

‘红颊’

‘Benihopp’

 ≥1.5或≤0.667 86 68
≥3.0或≤0.333 15 9

‘甜查理’

‘Sweet Charlie’

 ≥1.5或≤0.667 126 47
≥3.0或≤0.333 22 15
表1  差异表达磷酸化修饰肽段数统计
图3  ‘红颊’（A）和‘甜查理’（B）差异磷酸化肽段的保守域分析
图4  差异磷酸化蛋白的GO功能归类分析
图5  ‘红颊’（A）和 ‘甜查理’（B）响应炭疽菌侵染的差异磷酸化蛋白的KEGG通路富集分析
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