• 中文核心期刊
  • CSCD来源期刊
  • 中国科技核心期刊
  • CA、CABI、ZR收录期刊

稻瘟病菌MoDock1和MoElmo1蛋白的功能分析

Pathogenic Functions of MoDock1 and MoElmo1 in Magnaporthe oryzae

  • 摘要:
    目的 稻瘟病菌(Magnaporthe oryzae)是引起水稻稻瘟病的病原菌,探究其功能基因以揭示致病分子机理。
    方法 通过生物信息学方法鉴定稻瘟病菌中Dock180和ELMO的同源蛋白,对其编码基因进行敲除并对获得的基因缺失突变体进行表型分析,再通过免疫共沉淀试验分析二者的互作关系。
    结果 稻瘟病菌中鉴定得到MoDOCK1MoELMO1基因,并获得各自基因缺失突变体,两种突变体具有几乎一致的表型。MoDock1、MoElmo1的缺失影响了稻瘟病菌分生孢子的粘着,造成芽管变长且有隔膜,附着胞形成延缓。加入8-Br-cAMP或IBMX可以修复附着胞形成过程中的缺陷,表明MoDock1、MoElmo1可能在稻瘟病菌cAMP-PKA信号途径中发挥功能。洋葱表皮侵染试验结果表明基因缺失突变体的侵染能力有所下降。水稻接种显示,MoDock1、MoElmo1的缺失导致稻瘟病菌的致病性减弱。最后,通过免疫共沉淀证明,MoDock1和MoElmo1存在相互作用。
    结论 MoDock1和MoElmo1存在相互作用,二者是稻瘟病菌分生孢子粘着能力、附着胞正常形成和完整致病性所必需的。

     

    Abstract:
    Objective Roles of the genes associated with MoDock1 and MoElmo1 proteins in Magnaporthe oryzae played in rice blast disease were studied.
    Methods  Homologies of Dock180 and ELMO in M. oryzae were identified by bioinformatics method, and the coding genes knocked out to create mutants for determinations of phenotype and interactions between the two proteins by a co-immunoprecipitation test.
    Results  MoDOCK1 and MoELMO1 were identified in M. oryzae and deleted to obtain the mutants that were almost identical in phenotype. Absence of the two proteins in the mutants significantly affected the conidial adhesion resulting in long bud tubes with septum and delayed the appressoria formation. Since the defective appressoria formation could be repaired by the addition of 8-Br-cAMP or IBMX, it suggested that MoDock1 and MoElmo1 might be involved in the cAMP-PKA signaling pathway. To test that hypothesis, onion epidermis was infected with the mutants to show a declined infectious ability, and on a rice plant, a lowered pathogenicity. Furthermore, the co-immunoprecipitation experiment confirmed the MoDock1-MoElmo1 interaction as well.
    Conclusion MoDock1 and MoElmo1 in M. oryzae provided the conidia adhesion and appressoria formation essential for the pathogenicity of the pathogen on rice.

     

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