Rac1 is required for pathogenicity and Chm1-dependent conidiogenesis in rice fungal pathogen Magnaporthe grisea.

Rac1 is required for pathogenicity and Chm1-dependent conidiogenesis in rice fungal pathogen Magnaporthe grisea.
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水稻真菌病原菌稻瘟病菌的致病性和 Chm1 依赖性分生孢子发生需要 Rac1

DOI:
10.1371/journal.ppat.1000202
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发表时间:
2008-11
期刊:
影响因子:
6.7
通讯作者:
Wang, Zonghua
Wang, Zonghua
中科院分区:
医学1区
文献类型:
--
作者:
Chen, Jisheng;Zheng, Wu;Zheng, Shiqin;Zhang, Dongmei;Sang, Weijian;Chen, Xiao;Li, Guangpu;Lu, Guodong;Wang, Zonghua

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Rac1 是一种小型 GTP 酶,参与许多生物体中的肌动蛋白细胞骨架组织和极化细胞生长。在这项研究中,我们研究了 MgRac1(稻瘟病菌中 Rac1 同源物)的生物学功能。 MgRac1 缺失突变体在分生孢子产生方面存在缺陷。在产生的少数分生孢子中,它们是畸形的并且在附着形成方面有缺陷,因此失去了致病性。与天然 MgRac1 的基因互补可完全恢复所有这些缺陷表型。一致地,MgRac1 显性失活等位基因的表达表现出与缺失突变体相同的缺陷,而 MgRac1 组成型活性等位基因的表达可以诱导异常大的分生孢子,其感染相关生长存在缺陷。此外,我们通过酵母双杂交测定显示了 MgRac1 与其效应子(包括 PAK 激酶 Chm1 和 NADPH 氧化酶(Nox1 和 Nox2))之间的相互作用。虽然 Nox 蛋白对于致病性很重要,但 MgRac1-Chm1 相互作用负责分生孢子发生。一个组成型活性 Chm1 突变体,其中 Rac1 结合 PBD 结构域被去除,完全恢复了 MgRac1 缺失突变体的分生孢子形成,但这些分生孢子不能正常发育附着胞,并且对水稻植物不致病。我们的数据表明 MgRac1-Chm1 途径负责分生孢子发生,但其他途径(包括 Nox 途径)对于附着细胞的形成和致病性是必需的。
Rac1 is a small GTPase involved in actin cytoskeleton organization and polarized cell growth in many organisms. In this study, we investigate the biological function of MgRac1, a Rac1 homolog in Magnaporthe grisea. The MgRac1 deletion mutants are defective in conidial production. Among the few conidia generated, they are malformed and defective in appressorial formation and consequently lose pathogenicity. Genetic complementation with native MgRac1 fully recovers all these defective phenotypes. Consistently, expression of a dominant negative allele of MgRac1 exhibits the same defect as the deletion mutants, while expression of a constitutively active allele of MgRac1 can induce abnormally large conidia with defects in infection-related growth. Furthermore, we show the interactions between MgRac1 and its effectors, including the PAK kinase Chm1 and NADPH oxidases (Nox1 and Nox2), by the yeast two-hybrid assay. While the Nox proteins are important for pathogenicity, the MgRac1-Chm1 interaction is responsible for conidiogenesis. A constitutively active Chm1 mutant, in which the Rac1-binding PBD domain is removed, fully restores conidiation of the MgRac1 deletion mutants, but these conidia do not develop appressoria normally and are not pathogenic to rice plants. Our data suggest that the MgRac1-Chm1 pathway is responsible for conidiogenesis, but additional pathways, including the Nox pathway, are necessary for appressorial formation and pathogenicity.
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