香蕉枯萎病菌应对外源氧化胁迫的分子作用机制研究

Fusarium wilt of banana, caused by Fusarium oxysporum f. sp. cubense (Foc), is a destructive disease that has caused the greatest damage to banana crops worldwide. To research the molecular basis of this disease not only help us to understand the pathogen-host plant interaction, but also provide new targets for exploiting new fungicide and ultimately control of banana wilt disease hazards. By taking Foc4 as research object, using technologies of RNA-Seq (Quantification), PEG-mediated protoplast transformation, ROS detection and real-time quantitative PCR, we will carry out research in the following aspects: ①Analyses of the differentially expressed genes between wild-type strain B2 of Foc4 which are induced by H2O2 and the control in early phase; ②To get transcription factor in differentially display genes and construct target gene deletion mutant in Foc4 B2 strain; ③To determine whether or not target gene deletion mutants are hypersensitive to H2O2; ④To investigate whether or not target genes mediate host-derived ROS detoxification through regulation of gene expression; ⑤To detect whether or not gene deletion mutants can cause oxidative burst in the roots of banana tissue-culture plantlets and attenuates the virulence. We expect to clarify molecular mechanism of response of Foc4 to host-derived oxidative stress.

香蕉枯萎病是一种对全世界香蕉生产造成巨大损失的植物病害，研究香蕉枯萎病菌的致病机理不但能了解病原菌与寄主互作的详细机制，而且对开发新的杀菌剂从而控制香蕉枯萎病的危害具有重要意义。本项目拟以香蕉枯萎病病原菌（Foc4）为研究对象，采用最新的数字基因表达谱升级版分析技术(RNA-Seq (Quantification))、PEG介导的原生质体转化技术、ROS检测技术和荧光定量PCR技术等进行如下研究：①外源H2O2诱导下的Foc4野生型B2菌株与对照的B2菌株在诱导早期不同时段的基因表达差异分析；②获得表达差异显著基因中的转录因子并构建其在野生型B2菌株中的缺失突变菌株；③检测突变菌株对外源ROS（H2O2）的敏感性；④检测突变菌株中与清除ROS相关的基因表达；⑤检测突变菌株感染香蕉组培苗后的活性氧迸发及突变菌株的致病能力。期望初步阐明病原菌抵抗香蕉苗寄主氧化胁迫的分子机制。

香蕉枯萎病是一种对全世界香蕉生产造成巨大损失的植物病害，研究香蕉枯萎病菌的致病机理不但能了解病原菌与寄主互作的详细机制，而且对开发新的杀菌剂从而控制香蕉枯萎病的危害具有重要意义。申请人的前期研究工作表明，香蕉枯萎病病原菌（尖孢镰刀菌古巴专化型4号生理小种，Foc4）在入侵香蕉苗根部早期，香蕉苗根部根部存在强烈的活性氧迸发现象，这表明病原菌与寄主互作过程中存在强烈的氧化胁迫现象。本项目在前期工作基础上，采用数字基因表达谱升级版分析技术(RNA-Seq(Quantification))，结合PEG介导的原生质体转化技术、ROS检测技术和荧光定量PCR等技术，研究了Foc4在外源氧化胁迫（H2O2）条件下的转录组水平的基因差异表达情况。研究了部分参与应对外源氧化胁迫的转录因子基因。初步阐述了Foc4应对外源氧化胁迫的分子机制。