甘蔗镰孢菌(<i>Fusarium sacchari</i>)金属蛋白酶类效应蛋白Fs03538参与调控植物免疫响应

doi:10.13926/j.cnki.apps.000921

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植物病理学报 ›› 2025, Vol. 55 ›› Issue (1) : 19-31. DOI: 10.13926/j.cnki.apps.000921

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细胞生物学、生理学、生物化学、分子生物学

甘蔗镰孢菌(Fusarium sacchari)金属蛋白酶类效应蛋白Fs03538参与调控植物免疫响应

王璐璐^1,2#, 吴登^1,2#, 洪天澍^1,2, 姚伟^1,2,3, 张木清^1,2,3*, 胡琴^1,2,3*

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Metalloproteinase effector Fs03538 of Fusarium sacchari is involved in regulation of plant immunity

WANG Lulu^1,2＃, WU Deng^1,2＃, HONG Tianshu^1,2, YAO Wei^1,2,3, ZHANG Muqing^1,2,3*, HU Qin^1,2,3*

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摘要

甘蔗镰孢菌(Fusarium sacchari)是引致甘蔗梢腐病的主要病原真菌之一。为明确F. sacchari中金属蛋白酶(metalloproteinase)类效应蛋白的功能,本研究前期利用F. sacchari基因组数据对其具有分泌性的金属蛋白酶基因进行预测,从中扩增得到一个锌依赖型金属蛋白酶类效应蛋白基因Fs03538,并对其功能进行了初步研究。结果显示:Fs03538含有一个典型的锌依赖型金属蛋白酶结构域(ZnMc super family),蛋白质N端的1-18个氨基酸序列含有特定的分泌信号序列,亚细胞定位显示Fs03538能够定位于植物细胞核中;qRT-PCR分析表明Fs03538在甘蔗镰孢菌侵染甘蔗12 h到达最高峰值,之后表达量持续维持在较高水平;根癌农杆菌(Agrobacterium tumefaciens)介导的烟草瞬时表达系统确定Fs03538可以抑制小鼠促凋亡蛋白BAX引起的烟草细胞坏死,在F. sacchari中敲除Fs03538后发现突变体在生长速度、产孢量与野生型无明显差异,但致病力显著下降。以上结果表明,Fs03538在F. sacchari入侵寄主的过程中高量表达,可能通过进入寄主细胞核中抑制寄主免疫响应,是F. sacchari重要的毒力因子。

Abstract

Fusarium sacchari is one of the major pathogenic fungus that cause sugarcane pokkah boeng. In order to explore the function of metalloproteinase effector proteins in F. sacchari, we used the genomic data of F. sacchari to predict the secretory metalloproteinase proteins, and successfully amplified a zinc-type metalloproteinase effector protein gene Fs03538. The results showed that Fs03538 contained a typical ZnMc super family domain, and the 1-18 amino acid sequences at the N-terminal of the protein contain specific signal sequences. Subcellular localization showed that Fs03538 could be localized in the nucleus of Nicotiana benthamiana; qRT-PCR analysis showed that the expression level of Fs03538 was induced and reached the highest peak at 12 h post F. sacchari infection. Agrobacterium tumefaciens mediated transient expression system confirmed that Fs03538 could inhibit the necrosis of tobacco cells induced by mouse Bcl-2-associated X protein (BAX). As compared with the wild type F. sacchari strain CNO-1, the Fs03538 knock-out mutant showed no significant difference on mycelial growth and conidia production, but the pathogenicity on sugarcane was significantly decreased. Taken together, the results of the study suggest that Fs03538 is an important virulence factor of F. sacchari, which highly expressed at the time of infection and could inhibit the host immune responses by entering the host cell nucleus.

导出引用

王璐璐, 吴登, 洪天澍, 姚伟, 张木清, 胡琴. 甘蔗镰孢菌(Fusarium sacchari)金属蛋白酶类效应蛋白Fs03538参与调控植物免疫响应[J]. 植物病理学报, 2025, 55(1): 19-31 https://doi.org/10.13926/j.cnki.apps.000921

WANG Lulu, WU Deng, HONG Tianshu, YAO Wei, ZHANG Muqing, HU Qin. Metalloproteinase effector Fs03538 of Fusarium sacchari is involved in regulation of plant immunity[J]. Acta Phytopathologica Sinica, 2025, 55(1): 19-31 https://doi.org/10.13926/j.cnki.apps.000921

中图分类号： S432.44

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