Cao Jia-Yi, Xu You-Ping, Cai Xin-Zhong
Institute of Biotechnology, College of Agriculture and Biotechnology, Zhejiang University, 866 Yu Hang Tang Road, Hangzhou 310058, China.
Centre of Analysis and Measurement, Zhejiang University, 866 Yu Hang Tang Road, Hangzhou 310058, China.
J Proteomics. 2016 Jun 30;143:265-277. doi: 10.1016/j.jprot.2016.03.006. Epub 2016 Mar 4.
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.
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.
由核盘菌引起的白霉病是重要油料作物甘蓝型油菜中最重要的病害之一。然而,甘蓝型油菜对核盘菌的防御机制尚不清楚。在本研究中,我们进行了比较定量蛋白质组学分析,以揭示甘蓝型油菜对核盘菌的防御机制。使用基于TMT标记的定量分析技术,分析了接种核盘菌野生型菌株1980和非致病突变菌株Ep-1PB以及空琼脂块作为对照的甘蓝型油菜叶片的蛋白质组。分别在接种Ep-1PB和模拟接种的叶片、接种1980和模拟接种的叶片以及接种1980和Ep-1PB的叶片之间,共鉴定出79、299和173种持续差异表达的蛋白质。通过qRT-PCR分析证实了12种选定蛋白质的差异表达。基因本体论(GO)、京都基因与基因组百科全书(KEGG)和蛋白质-蛋白质相互作用预测分析表明,氧化还原稳态、脂质信号传导、钙信号传导、组蛋白和DNA甲基化介导的转录调控以及防御相关蛋白如防御素、防御素样蛋白和氰酸裂解酶,有助于甘蓝型油菜对核盘菌的防御。我们的结果为可能参与甘蓝型油菜对核盘菌防御反应的分子机制提供了新的见解。
核盘菌白霉病是重要油料作物甘蓝型油菜中最重要的病害之一。然而,甘蓝型油菜对核盘菌的防御机制迄今为止仍大多未知。在本研究中,我们通过对接种核盘菌野生型菌株1980和非致病突变菌株Ep-1PB以及空琼脂块作为对照的甘蓝型油菜叶片蛋白质组进行基于TMT标记的比较定量分析,解决了这个问题。通过分别对在接种Ep-1PB和对照叶片之间、接种1980和对照叶片之间以及接种1980和Ep-1PB的叶片之间持续差异表达的79、299和173种蛋白质进行比较分析,我们发现氧化还原稳态、脂质信号传导、钙信号传导、组蛋白和DNA甲基化介导的转录调控以及防御相关蛋白如防御素、防御素样蛋白以及氰酸裂解酶,有助于甘蓝型油菜对核盘菌的防御。值得注意的是,脂质信号传导、钙信号传导、组蛋白和DNA甲基化介导的转录调控以及氰酸裂解酶在甘蓝型油菜对核盘菌防御中的潜在作用此前未被报道,而是在本研究中首次揭示。本研究代表了对甘蓝型油菜接种核盘菌后蛋白质谱的最广泛分析,并且首次包括了接种野生型菌株和核盘菌非致病突变菌株的植物之间比较的结果。总体而言,我们的结果为甘蓝型油菜与核盘菌相互作用的分子机制提供了新的见解。