多组学技术揭示葡萄叶片响应灰葡萄孢菌侵染的抗性机制
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浙江大学学报（农业与生命科学版） 45（3）：306 ~316，2019
Journal of Zhejiang University (Agric. & Life Sci.) http：//www.zjujournals.com/agr
E-mail：zdxbnsb@zju.edu.cn
DOI：10.3785/j.issn.1008-9209.2018.11.121
Key words grape; Botrytis cinerea; proteomics; metabolomics; salicylic acid
基金项目：国家葡萄产业技术体系项目（CARS-29-10）。 ∗通信作者（Corresponding author）：蒋爱丽（https://orcid.org/0000-0003-1183-5655），E-mail：putaojal@163.com 第一作者（First author）：方献平（https://orcid.org/0000-0001-7461-9170），E-mail：fxpbio@163.com 收稿日期（Received）：2018-11-12；接受日期（Accepted）：2019-01-29
FANG Xianping, HE Yani, XI Xiaojun, ZHA Qian, ZHANG Liqing, JIANG Aili* (Institute of Forestry and Pomology, Shanghai Academy of Agricultural Sciences, Shanghai 201403, China)
关键词 葡萄；灰葡萄孢菌；蛋白质组学；代谢组；水杨酸
中图分类号 S 663.1
文献标志码 A
Multi-omics reveals the resistance mechanism of grape leaves in response to Botrytis cinerea. Journal of Zhejiang UΒιβλιοθήκη Baiduiversity (Agric. & Life Sci.), 2019,45(3):306-316
Abstract Liquid chromatography and mass spectrometry based label-free proteomics and non-target metabolomics technology were used to study the proteome and metabolome change of disease-resistant grape cultivar‘Shenfeng’infected with Botrytis cinerea. There were 1 374 proteins and 33 metabolites showing more than 1.5-fold changes in‘Shenfeng’leaves infected with B. cinerea, respectively. The differentially expressed proteins and metabolites were analyzed by gene ontology annotation and bioinformatics. The results showed that B. cinerea infection changed the expression level of chloroplast proteins, and mainly focused on plant and pathogen interaction, synthesis pathways of plant hormones and alkaloids. Multi-omics analysis further showed that there was a consistent increase in the expression levels of chorismic acid, salicylic acid, isochorismic pyruvate lyase pchB, transcription factor TGA and pathogenesis-related protein PR-1 in the salicylic acid-mediated disease-resistant signal transduction pathway. The full activation of salicylic acid-mediated disease resistance signaling pathway is an effective means for grape leaves to resist B. cinerea infection. The result is of great benefit to further deeply reveal the molecular mechanisms of plant-pathogen interactions and the breeding of pathogen-resistant grape varieties.
多组学技术揭示葡萄叶片响应灰葡萄孢菌侵染的抗性机制
方献平，和雅妮，奚晓军，查倩，张丽勍，蒋爱丽*
（上海市农业科学院林木果树研究所，上海 201403）
摘要 以葡萄灰霉病高抗品种‘申丰’叶片为试验材料，采用基于液相色谱质谱联用的非标记定量蛋白质组学和 非靶向定量代谢组学技术，比较了叶片在灰葡萄孢菌侵染胁迫 3 d 后体内蛋白质和代谢物的差异变化水平。试验 结果表明，葡萄叶片中有 1 374 个蛋白质和 33 种小分子代谢物在病菌侵染后发生了 1.5 倍以上的差异表达（P＜ 0.05）。功能注释和代谢通路富集等生物信息学分析发现，灰葡萄孢菌侵染对叶绿体蛋白表达影响最大，且主要集 中在植病互作、植物激素与生物碱合成 3 条信号路径上。基于多组学数据的联合分析进一步表明，水杨酸合成与 信号转导通路中的分支酸、水杨酸、异分支酸丙酮酸裂解酶 pchB、转录因子 TGA 和病程相关蛋白 PR-1 表达水平 显著上调。水杨酸介导的抗病信号通路的全面激活是葡萄叶片抵御灰葡萄孢菌侵染的有效手段。本研究发现为 后续深入揭示葡萄灰霉病菌互作分子机制及葡萄抗病新品种选育奠定了理论研究基础。
Journal of Zhejiang University (Agric. & Life Sci.) http：//www.zjujournals.com/agr
E-mail：zdxbnsb@zju.edu.cn
DOI：10.3785/j.issn.1008-9209.2018.11.121
Key words grape; Botrytis cinerea; proteomics; metabolomics; salicylic acid
基金项目：国家葡萄产业技术体系项目（CARS-29-10）。 ∗通信作者（Corresponding author）：蒋爱丽（https://orcid.org/0000-0003-1183-5655），E-mail：putaojal@163.com 第一作者（First author）：方献平（https://orcid.org/0000-0001-7461-9170），E-mail：fxpbio@163.com 收稿日期（Received）：2018-11-12；接受日期（Accepted）：2019-01-29
FANG Xianping, HE Yani, XI Xiaojun, ZHA Qian, ZHANG Liqing, JIANG Aili* (Institute of Forestry and Pomology, Shanghai Academy of Agricultural Sciences, Shanghai 201403, China)
关键词 葡萄；灰葡萄孢菌；蛋白质组学；代谢组；水杨酸
中图分类号 S 663.1
文献标志码 A
Multi-omics reveals the resistance mechanism of grape leaves in response to Botrytis cinerea. Journal of Zhejiang UΒιβλιοθήκη Baiduiversity (Agric. & Life Sci.), 2019,45(3):306-316
Abstract Liquid chromatography and mass spectrometry based label-free proteomics and non-target metabolomics technology were used to study the proteome and metabolome change of disease-resistant grape cultivar‘Shenfeng’infected with Botrytis cinerea. There were 1 374 proteins and 33 metabolites showing more than 1.5-fold changes in‘Shenfeng’leaves infected with B. cinerea, respectively. The differentially expressed proteins and metabolites were analyzed by gene ontology annotation and bioinformatics. The results showed that B. cinerea infection changed the expression level of chloroplast proteins, and mainly focused on plant and pathogen interaction, synthesis pathways of plant hormones and alkaloids. Multi-omics analysis further showed that there was a consistent increase in the expression levels of chorismic acid, salicylic acid, isochorismic pyruvate lyase pchB, transcription factor TGA and pathogenesis-related protein PR-1 in the salicylic acid-mediated disease-resistant signal transduction pathway. The full activation of salicylic acid-mediated disease resistance signaling pathway is an effective means for grape leaves to resist B. cinerea infection. The result is of great benefit to further deeply reveal the molecular mechanisms of plant-pathogen interactions and the breeding of pathogen-resistant grape varieties.
多组学技术揭示葡萄叶片响应灰葡萄孢菌侵染的抗性机制
方献平，和雅妮，奚晓军，查倩，张丽勍，蒋爱丽*
（上海市农业科学院林木果树研究所，上海 201403）
摘要 以葡萄灰霉病高抗品种‘申丰’叶片为试验材料，采用基于液相色谱质谱联用的非标记定量蛋白质组学和 非靶向定量代谢组学技术，比较了叶片在灰葡萄孢菌侵染胁迫 3 d 后体内蛋白质和代谢物的差异变化水平。试验 结果表明，葡萄叶片中有 1 374 个蛋白质和 33 种小分子代谢物在病菌侵染后发生了 1.5 倍以上的差异表达（P＜ 0.05）。功能注释和代谢通路富集等生物信息学分析发现，灰葡萄孢菌侵染对叶绿体蛋白表达影响最大，且主要集 中在植病互作、植物激素与生物碱合成 3 条信号路径上。基于多组学数据的联合分析进一步表明，水杨酸合成与 信号转导通路中的分支酸、水杨酸、异分支酸丙酮酸裂解酶 pchB、转录因子 TGA 和病程相关蛋白 PR-1 表达水平 显著上调。水杨酸介导的抗病信号通路的全面激活是葡萄叶片抵御灰葡萄孢菌侵染的有效手段。本研究发现为 后续深入揭示葡萄灰霉病菌互作分子机制及葡萄抗病新品种选育奠定了理论研究基础。