GPI anchoring controls cell wall integrity, immune evasion and surface localization of ChFEM1 for infection of Cochlibolus heterostrophus

doi:10.1016/j.jia.2024.09.033

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GPI anchoring controls cell wall integrity, immune evasion and surface localization of ChFEM1 for infection of Cochlibolus heterostrophus

Hong Hu^1*, Tiangu Liu^1*, Xinyun Xie¹, Fuyan Li¹, Caiyun Liu², Jintao Jiang¹, Zhigang Li³, Xiaolin Chen^1#

¹State Key Laboratory of Agricultural Microbiology and Provincial Key Laboratory of Plant Pathology of Hubei Province, College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, China

²Guangdong Provincial Key Laboratory of High Technology for Plant Protection, Plant Protection Research Institute, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, China

³Key Laboratory of Green Prevention and Control of Tropical Plant Diseases and Pests, Ministry of Education, College of Plant Protection, Hainan University, Haikou 570000, China

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摘要

GPI（糖基磷脂酰肌醇）锚定修饰在不同物种中发挥着粘附、分子吸收、信号转导和保护、细胞壁组分的合成和重组以及细胞壁蛋白的锚定等作用，且对真菌的生长发育和致病过程发挥着重要作用。目前关于 GPI 锚定修饰在真菌中虽然已有了初步研究，但其生物学功能和致病调控机制尚需深入揭示。本文对玉米小斑病菌 (Cochlibolus heterostrophus) GPI 锚定修饰的生物学功能和致病机制进行研究。首先，探究了 GPI 锚生物合成通路关键蛋白 ChGPI7 在玉米小斑病菌生长发育和致病过程中的功能及其参与致病的调控机制。结果表明，GPI锚定蛋白在玉米小斑菌的菌丝、附着胞和侵染菌丝中广泛表达。玉米小班菌ChGPI7基因的缺失会导致玉米小斑菌生长速度和分生孢子产孢能力显著下降。同时玉米小班菌ChGPI7基因敲除体∆Chgpi7的附着胞形成和侵染过程明显受阻，导致其致病力显著下降。∆Chgpi7敲除体还表现出严重的细胞壁完整性缺陷。ChGPI7基因的缺失和HF pyridine处理都会导致玉米小斑菌的细胞壁蛋白丰度减弱和几丁质的暴露，说明GPI锚定蛋白可以保护几丁质不被宿主免疫识别。在玉米小斑菌中，共有124个蛋白质被预测为GPI锚定修饰蛋白，其中包括一种假定的细胞壁糖蛋白ChFEM1，并发现其功能受 GPI 锚定修饰调控。缺失ChFEM1也会导致玉米小斑菌的致病力显著下降、侵染结构和细胞壁完整性的缺陷。我们的研究进一步发现，在玉米小斑菌中，ChGPI7调控ChFEM1蛋白的细胞壁定位和蛋白丰度。综合以上结果可以表明，GPI锚定修饰调节了玉米小斑菌的细胞壁完整性，免疫逃避和ChFEM1蛋白的细胞壁定位。研究结果为深入研究玉米小斑病菌的致病机制提供了理论基础，同时为开发防治玉米小斑病菌的新型药剂提供了潜在的靶标。

Abstract

Glycosylphosphatidylinositol (GPI) anchoring is one of the common post-translational modifications in eukaryotic cells. In fungi, it exerts a wide range of biological functions by targeting proteins to the cell wall, but only few studies focus on the roles of GPI anchoring in plant pathogenic fungi. Here, we reveal a role of GPI anchoring in the maize fungal pathogen Cochlibolus heterostrophus. We found that GPI-anchored proteins were widely accumulated in hyphae, appressorium and infection hyphae of C. heterostrophus. Deletion of ChGPI7, which encodes a key enzyme involved in the biosynthesis of GPI anchors, resulted in significant reduction of vegetative growth and conidiation, as well as virulence due to impairment of appressorium formation and invasive growth. The ∆Chgpi7 mutants also showed severe defects in cell wall integrity, resulting in a significant reduction of stress resistance. Deletion of ChGPI7 and hydrofluoric acid (HF) pyridine treatment both led to removal of cell wall GPI-anchored proteins and exposure of chitin, the results suggested that GPI anchored proteins could protect chitin from host immune recognition. A total of 124 proteins were predicted to be GPI anchored proteins in C. heterostrophus, including a putative cell wall glycoprotein ChFEM1. Deletion of ChFEM1 also resulted in significant reduction in virulence and defects in infection structures, as well as cell wall integrity. We further found that cell wall localization and protein abundance of ChFEM1 were affected by ChGPI7. Our results showed that GPI anchoring regulates cell wall integrity and immune evasion for infection of C. heterostrophus.

Keywords: Cochlibolus heterostrophus GPI anchor immune evasion cell wall integrity cell wall protein fungal infection

Online: 26 September 2024

Fund:

This work was supported by the Fundamental Research Funds for the Central Universities, China (2021ZKPY007).

About author: #Correspondence Xiaolin Chen, E-mail: chenxiaolin@mail.hzau.edu.cn *These authors contribute equal to this work.

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Hong Hu, Tiangu Liu, Xinyun Xie, Fuyan Li, Caiyun Liu, Jintao Jiang, Zhigang Li, Xiaolin Chen. 2024. GPI anchoring controls cell wall integrity, immune evasion and surface localization of ChFEM1 for infection of Cochlibolus heterostrophus. Journal of Integrative Agriculture, Doi:10.1016/j.jia.2024.09.033

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