茭白对菰黑粉菌侵染的应激防御反应研究

Zizania latifolia is the second aquatic vegetable of china, which had high economic benefits. The edible stem of Z. latifolia is a morbid organs caused by Ustiligo esculenta infection. The defense action played an important role in the gall formation during the interactions of U. esculenta and Z. latifolia. Researches about related gene actions had not been reported about the interactions of U. esculenta and Z. latifolia. Through the 2-DE analysis of Z. latifolia stem, differential expression of some disease-resistance related proteins had been found between different developmental stage, for example the RPR1 which could be induced by many fungi pathogens. In this research, some disease-resistance related genes would be cloned from cultivated Z. latifolia. The expression of these genes would be researched among cultivated Z. latifolia of gall formation normally, cultivated Z. latifolia of gall formation un-normally and cultivated Z. latifolia with no gall formation. After localized expression analysis, the regulation of genes expression in the defense action during gall formation of Z. latifolia would be clarified and farther validated through the experiments of infection between Z. latifolia and U. esculenta. Besides, the patterns of gene expression would be discussed .All of the above researches would provide more theory basis for the mechanism of gall formation in Z. latifolia.

茭白是我国第二大水生蔬菜，经济效益很高。茭白肉质茎是由菰黑粉菌侵染茭白植株后产生的病态器官，茭白对菰黑粉菌侵染的应激防御反应在茭白孕茭具有重要作用。目前尚无茭白植株与菰黒粉菌侵染互作相关基因的研究。.本课题组通过不同发育阶段茭白茎部蛋白质组的比较分析，发现肉质茎形成期间茎部对菰黒粉菌具有明显的感病反应，存在与多种病原真菌诱导表达相关的广谱性抗病相关蛋白RPR1等的差异表达。.本项目拟克隆正常栽培茭白中抗病相关基因；筛选正常茭白、雄茭及灰茭之间差异表达的抗病相关基因并进行定位表达分析，阐明抗病相关基因在茭白孕茭应激防御中的表达调控；通过互作分析明确抗病相关基因对菰黒粉菌侵染的诱导响应；比较基因表达模式变化在孕茭表达调控中的作用，为进一步探讨茭白孕茭相关机制的研究提供理论依据。

茭白茎部膨大孕茭是由菰黑粉菌侵染茭白植株共同作用的结果，本项目对茭白茎部菰黑粉菌的生长分布进行了比较分析，阐明了不同茭白表型间菰黑粉菌的生长分布特点，揭示了茭白孕茭期间茎部菰黑粉菌生长的动态。基于茭白茎部膨大前后的蛋白质组分析，筛选鉴定了与茭白茎部膨大响应相关的差异表达蛋白，阐明了茭白茎部膨大相关的主要响应途径及响应时期。通过茎部膨大发育过程中的表达谱分析，阐明了与茭白茎部膨大相关的茭白植物与菰黑粉菌基因的响应时期，发现茭白茎部膨大前与膨大后间的响应基因最多。克隆获得了茭白Zl-RPM1和Zl-ADR基因全长序列，阐明了其在茭白孕茭表型和膨大期间茎部的表达模式；通过喷施杀真菌剂抑制茭白茎部菰黑粉菌生长，延迟了茭白茎部的膨大时间，分析了3个抗病相关基因在茭白延迟结茭期间的表达模式变化，揭示了菰黑粉菌侵染生长对Zl-RPM1基因表达的诱导调节关系。本项目通过对茭白茎部膨大相关的响应基因及蛋白的分析，结合茭白茎部菰黑粉菌侵染生长的动态分布，探讨了茭白对菰黑粉菌侵染生长的免疫防御调节机制。