TMT-based quantitative proteomics analyses reveal novel defense mechanisms of Brassica napus against the devastating necrotrophic pathogen Sclerotinia sclerotiorum
TMT-based quantitative proteomics analyses reveal novel defense mechanisms of Brassica napus against the devastating necrotrophic pathogen Sclerotinia sclerotiorum
复制标题
基于 TMT 的定量蛋白质组学分析揭示了甘蓝型油菜对抗毁灭性坏死性病原体核盘菌的新防御机制
DOI：
10.1016/j.jprot.2016.03.006
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发表时间：
2016-06-30
影响因子：
3.3
通讯作者：
Cai, Xin-Zhong
中科院分区：
文献类型：
--
作者：
Cao, Jia-Yi;Xu, You-Ping;Cai, Xin-Zhong
油菜白色霉病是由油菜核盘菌（Sclerotinia sclerotiorum）引起的重要病害之一。然而，B. napus对S.对核盘菌的了解很少。在这项研究中，我们进行了比较定量蛋白质组学分析，以揭示B。napus对S.菌核B.油菜叶片接种S.利用TMT标记的定量分析技术对核盘菌野生型菌株1980和非致病突变菌株Ep-1 PB以及作为对照的空琼脂塞进行了分析。共79，299和173个蛋白质始终差异表达之间的Ep-1 PB和模拟接种的叶子，1980年和模拟接种的叶子，以及1980年和Ep-1 PB接种的叶子，分别进行了鉴定。通过qRT-PCR分析证实了12种选择的蛋白质的差异表达。基因本体论（GO）、京都基因和基因组百科全书（KEGG）和蛋白质相互作用预测分析表明，氧化还原稳态、脂质信号、钙信号、组蛋白和DNA甲基化介导的转录调控以及防御素、防御素样蛋白和氰酸裂解酶等防御相关蛋白参与了S.菌核我们的研究结果为可能参与B防御反应的分子机制提供了新的见解。napus为S.意义：油菜菌核病（Sclerotinia白色mold disease）是重要油料作物甘蓝型油菜（Brassica napus）上最重要的病害之一。然而，B. napus对S.核盘菌至今仍在很大程度上未知。在这项研究中，我们通过对B的蛋白质组进行基于TMT标记的比较定量分析来解决这个问题。油菜叶片接种S.以核盘菌野生型菌株1980和非致病性突变菌株Ep-1 PB以及空琼脂塞为对照。通过对接种Ep-1 PB的叶片与对照叶片、接种1980的叶片与对照叶片以及接种1980的叶片与接种Ep-1 PB的叶片之间分别差异表达的79、299和173个蛋白质进行比较分析，我们揭示了氧化还原稳态、脂质信号、钙信号、组蛋白和DNA甲基化介导转录调节和防御相关蛋白如防御素和防御素样蛋白以及氰酸裂解酶对B有贡献。napus对S.菌核值得注意的是，脂质信号，钙信号，组蛋白和DNA甲基化介导的转录调控和氰酸裂解酶在B。napus对S.核盘菌是以前没有报道过的，而是在这项研究中首次公布。目前的研究代表了对B蛋白质谱的最广泛的分析。napus对S. sclerotiorum接种，并包括第一次与野生型菌株和S.菌核总的来说，我们的研究结果提供了新的见解之间的相互作用的分子机制B。油菜和S.菌核(C)© 2016 Elsevier B. V.版权所有。
The white mould disease, caused by Sclerotinia sclerotiorum, is one of the most important diseases in the vital oil crop Brassica napus. Nevertheless, the defense mechanisms of B. napus against S. sclerotiorum are poorly understood. In this study, we performed comparative quantitative proteomics analyses to reveal B. napus defense mechanisms against S. sclerotiorum. The proteomes of B. napus leaves inoculated with S. sclerotiorum wild-type strain 1980 and nonpathogenic mutant strain Ep-1PB as well as empty agar plug as the control were analyzed using TMT label-based quantitative analysis technique. A total of 79, 299 and 173 proteins consistently differentially expressed between Ep-1PB- and mock-inoculated leaves, 1980- and mock-inoculated leaves, as well as 1980- and Ep-1PB-inoculated leaves, respectively, were identified. The differential expression of 12 selected proteins was confirmed by qRT-PCR analyses. The Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) and protein-protein interaction prediction analyses revealed that redox homeostasis, lipid signaling, calcium signaling, histone and DNA methylation-mediated transcription regulation and defense-related proteins such as defensin and defensin-like proteins and cyanate lyase, contribute to defense against S. sclerotiorum. Our results provide new insights into molecular mechanisms that may be involved in defense responses of B. napus to S. sclerotiorum.Significance: The Sclerotinia white mould disease is one of the most important diseases in the significant oil crop Brassica napus. Nevertheless, the defense mechanisms of B. napus against S. sclerotiorum are still largely unknown to date. In this study, we addressed this issue by performing TMT label-based comparative quantitative analyses of the proteomes of B. napus leaves inoculated with S. sclerotiorum wild-type strain 1980 and nonpathogenic mutant strain Ep-1PB as well as empty agar plug as the control. Through comparative analyses on 79, 299, and 173 proteins that are consistently differentially expressed in between Ep-1PB-inoculated and the control leaves, 1980-inoculated and the control leaves, as well as 1980-inoculated and Ep-1PB-inoculated leaves, respectively, we revealed that redox homeostasis, lipid signaling, calcium signaling, histone and DNA methylation-mediated transcription regulation and defense-related proteins such as defensin and defensin-like proteins as well as cyanate lyase, contribute to B. napus defenses against S. sclerotiorum. Notably, the potential role of lipid signaling, calcium signaling, histone and DNA methylation-mediated transcription regulation and cyanate lyase in B. napus defense against S. sclerotiorum are not reported previously but rather unveiled for the first time in this study. The current study represents the most extensive analysis of the protein profile of B. napus in response to S. sclerotiorum inoculation and includes for the first time the results from comparison between plants inoculated with the wild-type strain and a nonpathogenic mutant strain of S. sclerotiorum. Collectively, our results provide new insights into the molecular mechanisms of interactions between B. napus and S. sclerotiorum. (C) 2016 Elsevier B.V. All rights reserved.