Journal of Integrative Agriculture ›› 2023, Vol. 22 ›› Issue (11): 3444-3457.DOI: 10.1016/j.jia.2023.04.005

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FgGyp8通过调节FgSnc1介导的分泌囊泡与质膜融合影响禾谷镰刀菌的生长发育和致病性

  

  • 收稿日期:2022-11-02 接受日期:2023-02-19 出版日期:2023-11-20 发布日期:2023-11-09

FgGyp8 as a putative FgRab1 GAP is required for growth and pathogenesis by regulating FgSnc1-mediated secretory vesicles fusion in Fusarium graminearum

ZHANG Xing-zhi1, 2, CHEN Shuang2, Yakubu Saddeeq ABUBAKAR2, 3, MAO Xu-zhao1, 2, MIAO Peng-fei2, WANG Zong-hua1, 2, ZHOU Jie2#ZHENG Hua-wei1#   

  1. 1 Fujian Key Laboratory on Conservation and Sustainable Utilization of Marine Biodiversity/Ministerial and Provincial Joint Innovation Centre for Safety Production of Cross-Strait Crops, Minjiang University, Fuzhou 350108, P.R.China
    2 Fujian University Key Laboratory for Plant-Microbe Interaction, College of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, P.R.China
    3 Department of Biochemistry, Faculty of Life Sciences, Ahmadu Bello University, Zaria 810211, Nigeria
  • Received:2022-11-02 Accepted:2023-02-19 Online:2023-11-20 Published:2023-11-09
  • About author:ZHANG Xing-zhi, E-mail: 731563976@qq.com; #Correspondence ZHENG Hua-wei, E-mail: zhw@mju.edu.cn; ZHOU Jie, E-mail: jiezhou@fafu.edu.cn
  • Supported by:
    This research was funded by the National Natural Science Foundation of China (31970141), the Natural Science Foundation of Fujian Province, China (2020J06047), the Foundation of Minjiang University, China (MJY19019), and the Foundation of Fujian Agriculture and Forestry University, China (KFb22050XA).

摘要:

禾谷镰刀菌是一种重要的植物病原真菌,可引起小麦、大麦等多种禾谷类农作物致病及减产。模式真菌酿酒酵母中Gyp8蛋白能水解GTP激活态的Ypt1Rab1)然而,在植物病原真菌中Gyp8同源蛋白的功能仍然未知本研究从遗传学和病理学的角度对禾谷镰刀菌中Gyp8同源蛋白FgGyp8的功能进行研究。通过基因敲除和表型分析,我们发现FgGyp8是禾谷镰刀菌营养菌丝生长所必需。突变体ΔFggyp8分生孢子产量、大小隔膜数与野生型PH-1相比显著下降进一步发现FgGyp8禾谷镰刀菌小麦胚及麦穗的致病性起重要作用FgGyp8包含一个保守TBC(Tre2-Bub2-Cdc16)结构域结构域缺失结果表明,FgGyp8TBC结构域、C端和N末端对其生物学功能均有重要调控作用体外水解酶活性实验结果显示FgGyp8是FgRab1的GTP酶激活蛋白(GAP,GTPase-activating protein)。此外,FgGyp8对FgSnc1蛋白介导的分泌囊泡与质膜的融合过程是必需的。最后,我们发现FgGyp8禾谷镰刀菌FgRab1的另一个GAP FgGyp1存在功能冗余。综上所述,本研究表明FgGyp8作为FgRab1GAP禾谷镰刀菌营养菌丝生长分生孢子形态建成、致病性所必需的。

Abstract:

Fusarium graminearum is an important plant pathogenic fungus that causes disease and yield reduction in many cereal crops, such as wheat and barley.  Gyp8 stimulates GTP hydrolysis on Ypt1 in yeast.  However, the functions of Gyp8 in plant pathogenic fungi are still unknown.  In this study, we investigated the roles of FgGyp8 in Fgraminearum by genetic and pathological analyses.  Through gene knockout and phenotypic analyses, we found that FgGyp8 is required for vegetative growth in Fgraminearum.  The conidiation, conidial size and number of septa per conidium of ΔFggyp8 mutant are significantly reduced when compared to the wild type PH-1.  Furthermore, FgGyp8 is crucial for pathogenicity on wheat coleoptiles and wheat heads.  FgGyp8 contains a conserved TBC domain.  Domain deletion analysis showed that the TBC domain, C- and N-terminal regions of FgGyp8 are all important for its biological functions in Fgraminearum.  Moreover, we showed that FgGyp8 catalyzes the hydrolysis of the GTP on FgRab1 to GDP in vitro, indicating that FgGyp8 is a GTPase-activating protein (GAP) for FgRab1.  In addition, we demonstrated that FgGyp8 is required for FgSnc1-mediated fusion of secretory vesicles with the plasma membrane in Fgraminearum.  Finally, we showed that FgGyp8 has functional redundancy with another FgRab1 GAP, FgGyp1, in Fgraminearum.  Taken together, we conclude that FgGyp8 is required for vegetative growth, conidiogenesis, pathogenicity and acts as a GAP for FgRab1 in Fgraminearum.

Key words: Fusarium graminearum ,  FgGyp8 ,  GTPase-activating protein ,  FgRab1 ,  conidiogenesis ,  pathogenicity