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第二信使c-di-GMP受体DGC3介导的内生蜡样芽孢杆菌生物防治小麦纹枯病的作用机制

第二信使c-di-GMP受体DGC3介导的内生蜡样芽孢杆菌生物防治小麦纹枯病的作用机制
  • 导航:首页 > 科学基金
  • 批准号:31572047
  • 批准年度: 2015年
  • 学科分类:植物病害生物防治(C140601) |
  • 项目负责人:王刚
  • 负责人职称:教授
  • 依托单位:河南大学
  • 资助金额:65万元
  • 项目类别:面上项目
  • 研究期限:2016年01月01日 至 2019年12月31日
  • 中文关键词: c-di-GMP;DGC3;内生蜡样芽孢;防治;纹枯病
  • 英文关键词:Bacillus;biofilm;colonization;sharp eyespot of wheat;c-di-GMP

项目摘要

中文摘要

利用内生细菌防治病害,可以克服根际细菌生物防治中易出现的不稳定现象。研究生防作用的代谢调控网络,有助于揭示生防机制。c-di-GMP是广泛存在于细菌中的一种第二信使信号分子,参与调节biofilm形成、运动性等多种基本生物学过程。前期我们证明了生防菌蜡样芽孢杆菌B3-7中存在c-di-GMP代谢网络,蛋白DGC3作为c-di-GMP受体参与介导B3-7对于小麦纹枯病生防作用中由DGC3到SbnB的信号传递,但是DGC3的效应蛋白和生防途径的中间组分未知。本研究鉴定DGC3的效应调节蛋白,研究DGC3和效应调节蛋白的互作方式,解析效应蛋白在生防中的作用。在此基础上,利用获得的16个生防变异突变体,分离生防相关基因并进行功能分析,确定生防途径的中间组分。通过上位性分析,解析不同组分的时空顺序,揭示DGC3介导的信号通路组成和c-di-GMP信号在B3-7生物防治中的作用。

英文摘要

Sharp eyespot of wheat is an important soil-borne disease of wheat worldwide. Control measures against this disease available at present are inefficient due to lack of wheat cultivars resistant to sharp eyespot and no efficient fungicide available. Biocontrol of sharp eyespot, as an alternative measure, have been investigated intensively. Bacillus cereus can form endospores that are tolerant to heat and desiccation, offer several advantages over other bacteria..The second messenger c-di-GMP is a broadly conserved intracellular signaling molecule that is important for controlling biofilm formation, adhesion, motility, virulence in diverse bacterial species. The intracellular levels of c-di-GMP are modulated by the opposite activity of diguanylate cyclases (DGCs), which contain GGDEF domains, and phosphodiesterases (PDEs), which contain EAL or HD-GYP domains. The presence of a variety of genes coding for GGDEF, EAL or GGDEF-EAL proteins in the genome of many bacterial species indicates that bacteria regulate c-di-GMP turnover in an extremely sophisticated manner. How microbes integrate multiple input signals to regulate critical biological processes is still largely unclear. .B.cereus B3-7 is an endophytic bacteria, isolated from wheat roots, which have biocontrol capacity against sharp eyespot of wheat. We have revealed that a degenerated GGDEF protein called DGC3 functioned as a c-di-GMP receptor which involves in biocontrol to sharp eyespot. To elucidate the function of DGC3 in the process of biocontrol. We constructed dgc3 deletion mutant(Δdgc3)and identified total of 72 differential expression proteins between B3-7 and its Δdgc3 mutant based on 2D electrophoresis. All of the 72 protein encoding genes were deleted and biocontrol activity of 72 gene deletion mutants were tested indivually. We found SbnB involved in biocontrol activity. No protein interaction found between DGC3 and SbnB. It was suggested that there existed a metabolism pathway between the c-di-GMP receptor DGC3 and SbnB,which mediated biocontrol of B3-7 to sharp eyespot of wheat. Based on research results previously, we would carry out co-immunoprecipitation, pull-down assay and Bacterial two-hybrid to characterize response regulator protein which binds DGC3 and relay signals to cellular processes and impact biocontrol activity of B3-7. We then would conduct biocontrol-related genes isolation from 16 biocontrol variant mutants characterized previously. We would analyze gene function so as to ascertain components of biocontrol pathway from DGC3 to SbnB. Next ,we would analyze the metabolic circuits of biocontrol related factors involved and carry out multigene deletion to disclose temporal and spatial expression patterns of genes in the process of biocontrol. We would assay the stability of mRNA and its encoding proteins of biocontrol gene to reveal the regulation patter caused by c-di-GMP levels. All the research findings will lay a foundation of development biocontrol agents.

评估说明

    国家自然科学基金项目“第二信使c-di-GMP受体DGC3介导的内生蜡样芽孢杆菌生物防治小麦纹枯病的作用机制”发布于爱科学iikx,并永久归类于相关科学基金导航中,仅供广大科研工作者查询、学习、选题参考。国科金是根据国家发展科学技术的方针、政策和规划,以及科学技术发展方向,面向全国资助基础研究和应用研究,发挥着促进我国基础研究源头创新的作用。国科金的真正价值在于它能否为科学进步和社会发展带来积极的影响。

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