一种潜在的菌丝融合蛋白复合体作用于假禾谷镰孢的发育和致病性，并与细胞自噬相关蛋白互作

doi:10.1016/j.jia.2023.09.005

摘要/Abstract

摘要：

菌丝融合在子囊真菌中普遍存在，调控多种发育过程。但是菌丝融合在植物病原真菌中的生物功能及分子机制尚不明了。本研究基于假禾谷镰孢T-DNA插入突变体库的筛选，鉴定到一个菌丝融合蛋白FpHam-2。利用酵母双杂交方法发现FpHam-2与另外两个菌丝融合蛋白FpHam-3和FpHam-4直接互作，且其作为STRIPAK复合体的核心元件在真核生物中非常保守。利用假禾谷镰孢遗传转化分别获得FpHam-2、FpHam-3和FpHam-4缺失的突变体菌株。表型分析发现，三种突变体具有相似的生物学缺陷表型，与野生型和回补菌株相比，基因缺失突变体的菌丝生长速度减慢、不能发生菌丝融合、对细胞膜、细胞壁和氧化胁迫的耐受性降低，不能产生分生孢子，且致病性降低。推测FpHam-2、FpHam-3和FpHam-4可能作为一种蛋白复合体起作用。通过酵母双杂交方法进一步验证FpHam-2与细胞自噬相关蛋白FpAtg3、FpAtg28和FpAtg33直接互作。假禾谷镰孢饥饿诱导后，野生型和回补菌株有大量自噬体被MDC染料标记，而FpHam-2缺失突变体的细胞自噬明显减弱。综合上述结果，本研究阐明了菌丝融合蛋白FpHam-2、FpHam-3和FpHam-4在假禾谷镰孢生长、产孢和致病中的重要作用，并初步揭示了其作用的潜在分子机制。

Abstract:

Hyphal fusion (anastomosis) is a common process serving many important functions at various developmental stages in the life cycle of ascomycetous fungi. However, the biological roles and molecular mechanisms in plant pathogenic fungi were widely unknown. In this study, a hyphal fusion protein FpHam-2 was screened from a T-DNA insertion mutant library of Fusarium pseudograminearum, and FpHam-2 interacts with another 2 hyphal fusion protein homologues FpHam-3 and FpHam-4. Each of these 3 genes deletion mutant revealed in similar defective phenotypes compared with the WT and complemented strains, including reduction in growth rate, defects in hyphal fusion and conidiation, more sensitive for cell membrane, cell wall and oxidative stress responses, and decreased in virulence. The yeast two-hybrid assay was used to identify that FpHam-2 interacts with 3 autophagy-related proteins, including FpAtg3, FpAtg28 and FpAtg33. Furthermore, FpHam-2-deletion mutant showed decreased accumulation of autophagic bodies in hypha. In conclusion, FpHam-2, FpHam-3 and FpHam-4 have an essential role for hyphal fusion and regulating the growth, conidiation and virulence in F. pseudograminearum.

Key words: Fusarium pseudograminearum , hyphal fusion , FpHam-2 , virulence , autophagy-related proteins

Linlin Chen, Yixuan Shan, Zaifang Dong, Yake Zhang, Mengya Peng, Hongxia Yuan, Yan Shi, Honglian Li, Xiaoping Xing. 一种潜在的菌丝融合蛋白复合体作用于假禾谷镰孢的发育和致病性，并与细胞自噬相关蛋白互作[J]. Journal of Integrative Agriculture, 2024, 23(12): 4093-4106.

Linlin Chen, Yixuan Shan, Zaifang Dong, Yake Zhang, Mengya Peng, Hongxia Yuan, Yan Shi, Honglian Li, Xiaoping Xing. A potential hyphal fusion protein complex with an important role in development and virulence interacts with autophagy-related proteins in Fusarium pseudograminearum[J]. Journal of Integrative Agriculture, 2024, 23(12): 4093-4106.

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