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| Identification of proteins associated with Fusarium crown rot resistance in wheat using label-free quantification analysis |
JIN Jing-jing1, 2, DUAN Shuo-nan3, QI Yong-zhi1, 2, ZHEN Wen-chao2, 4, MA Jun3 |
1 College of Plant Protection, Hebei Agricultural University, Baoding 071001, P.R.China
2 State Key Laboratory of North China Crop Improvement and Regulation, Baoding 071001, P.R.China
3 College of Agronomy and Biotechnology, China Agricultural University, Beijing 100193, P.R.China
4 College of Agronomy, Hebei Agricultural University, Baoding 071001, P.R.China |
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摘要
小麦茎基腐病是由镰刀菌属真菌引起的一种世界性土传病害，近年来已严重威胁到我国小麦的安全生产。目前，对该病害的研究主要集中在抗源筛选、抗病位点发掘、接种前后转录组分析等层面。本研究首次采用非标定量蛋白质组学方法，分析比较了我国小麦中抗品种和高感品种在接种前后24、48和72小时后的蛋白质组变化。试验共检测到9234个蛋白质，发掘到783个在接种后不同时间点差异表达的蛋白。对这些差异蛋白的富集分析表明，茎基腐病早期侵染对几丁质代谢、氨基糖代谢、乙醛酸和二元酸代谢，丙氨酸、天冬氨酸和谷氨酸代谢，MAPK信号通路，苯丙素生物合成等重要生物学功能和代谢通路具有显著影响。进一步分析发现共有33种与防御、细胞壁形成和光合作用等功能相关的蛋白质在抗、感品种间接种后多个时间点的表达呈现不同趋势。例如，中抗品种中几丁质酶在接菌后三个时间点均显著上调，而感病品种中接种前后则无显著变化。该酶参与降解真菌细胞壁的重要成分几丁质，从而限制真菌的生长。其它蛋白如糖基转移酶、过氧化物酶等也在中抗品种中特异上调表达。前人报道中，这些蛋白参与了其他重要小麦病害如赤霉病、白粉病等侵染后的应答反应。在感病品种中，小檗碱桥酶等特异下调表达。qRT-PCR试验表明，编码几丁质酶、糖基转移酶、过氧化物酶等8个抗病相关蛋白的基因在抗、感材料间接种前后表达趋势与蛋白质组学结果基本一致。本研究为进一步解析小麦抗茎基腐病分子机制奠定了有力基础，为候选基因筛选提供了重要参考。
Abstract Fusarium crown rot (FCR), typically caused by Fusarium pseudograminearum, is a severe soil-borne disease that, in recent years, has become an emerging threat to Chinese wheat crops. For the first time in this study, we investigated and compared the proteomic characteristics of two Chinese wheat varieties (04 Zhong 36 and Xinmai 26) at 24, 48, and 72 h post-inoculation using label-free quantitative proteomic analysis. A total of 9 234 proteins were successfully quantified, of which 783 were differentially expressed after inoculation. These proteins were mainly involved in metabolic, single-organism, and cellular processes. Thirty-three proteins associated with defense, cell wall formation, photosynthesis, etc., showed consistently different expression between the two genotypes at multiple time points. In particular, chitinase, which degrades chitin in the fungal cell wall and limits fungal growth, was exclusively and consistently upregulated in 04 Zhong 36 across the three time points. Other proteins such as flavonoid O-methyltransferase, glycosyltransferase, and peroxidase were only upregulated in 04 Zhong 36, and proteins, including the berberine bridge enzyme and rubisco large subunit-binding protein, were specifically downregulated in Xinmai 26. The expression of transcripts encoding eight selected proteins through qRT-PCR analysis supported the proteomic profiles. Overall, the results of this study allow us to understand FCR resistance in wheat at the protein level. Some proteins and their corresponding genes may be useful resources for the genetic improvement of FCR resistance in wheat.
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Received: 17 July 2020
Accepted:
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| Fund: This research was supported by the National Key Research and Development Program of China (2017YFD0300906 and 2018YFD0300501), the National Natural Science Foundation of China (31872865), and the State Key Laboratory of North China Crop Improvement and Regulation. |
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Corresponding Authors:
Correspondence ZHEN Wen-chao, Tel: +86-312-7528996, E-mail: wenchao@hebau.edu.cn; MA Jun, Tel: +86-10-62734514, E-mail: Junma@cau.edu.cn
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| About author: JIN Jing-jing, E-mail: jingjing1809@yeah.net; |
Cite this article:
JIN Jing-jing, DUAN Shuo-nan, QI Yong-zhi, ZHEN Wen-chao, MA Jun.
2021.
Identification of proteins associated with Fusarium crown rot resistance in wheat using label-free quantification analysis . Journal of Integrative Agriculture, 20(12): 3209-3221.
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