FgFim, a key protein regulating resistance to the fungicide JS399-19, asexual and sexual development, stress responses and virulence in Fusarium graminearum.
FgFim, a key protein regulating resistance to the fungicide JS399-19, asexual and sexual development, stress responses and virulence in Fusarium graminearum.
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DOI：
10.1111/mpp.12108
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发表时间：
2014-06
影响因子：
4.9
通讯作者：
Zhitian Zheng;Tao Gao;Yu Zhang;Yiping Hou;Jian-xin Wang;Ming-guo Zhou
中科院分区：
文献类型：
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作者：
Zhitian Zheng;Tao Gao;Yu Zhang;Yiping Hou;Jian-xin Wang;Ming-guo Zhou
纤维蛋白是一种肌动蛋白成束蛋白，存在于肠道微绒毛、毛细胞静纤毛和成纤维细胞丝状伪足中。其同源物 Sac6p 已被证明在酿酒酵母的内吞作用和多种细胞过程中发挥着关键作用。通过筛选由 HPH-HSV-tk 盒介导的整合产生的突变体文库，鉴定出来自小麦赤霉病病原真菌禾谷镰刀菌菌株 Y2021A 的 FgFim，该菌株对杀菌剂 JS399-19 具有高度抗性。通过构建 FgFim 的缺失突变体（命名为 ΔFgFim-15）来评估 FgFim 的功能。缺失突变体表现出菌丝生长速率降低、分生孢子形成减少、分生孢子萌发延迟、菌丝形状不规则、高压灭菌的小麦籽粒缺乏有性繁殖以及对JS399-19的抗性显着降低。 ΔFgFim-15 还表现出对多种金属阳离子、诱导渗透应激和氧化应激的物质以及损伤细胞膜和细胞壁的物质的敏感性增加。致病性测定表明，FgFim 缺失突变体对开花小麦穗的毒力受损，这与其在宿主组织中毒素脱氧雪腐镰刀菌烯醇产量的减少相一致。所有这些缺陷都通过突变体与亲本 FgFim 基因的遗传互补得到恢复。实时定量聚合酶链式反应（PCR）分析表明，编码甾醇14α-去甲基酶的三个Cyp51基因的基础表达在突变体中显着低于亲本菌株。这项研究的结果表明，FgFim 在调节禾谷镰刀菌对 JS399-19 的抗性以及各种细胞过程中发挥着关键作用。
Fimbrin is an actin-bundling protein found in intestinal microvilli, hair cell stereocilia and fibroblast filopodia. Its homologue Sac6p has been shown to play a critical role in endocytosis and diverse cellular processes in Saccharomyces cerevisiae. FgFim from the wheat scab pathogenic fungus Fusarium graminearum strain Y2021A, which is highly resistant to the fungicide JS399-19, was identified by screening a mutant library generated by HPH-HSV-tk cassette-mediated integration. The functions of FgFim were evaluated by constructing a deletion mutant of FgFim, designated ΔFgFim-15. The deletion mutant exhibited a reduced rate of mycelial growth, reduced conidiation, delayed conidium germination, irregularly shaped hyphae, a lack of sexual reproduction on autoclaved wheat kernels and a dramatic decrease in resistance to JS399-19. ΔFgFim-15 also exhibited increased sensitivity to diverse metal cations, to agents that induce osmotic stress and oxidative stress, and to agents that damage the cell membrane and cell wall. Pathogenicity assays showed that the virulence of the FgFim deletion mutant on flowering wheat heads was impaired, which was consistent with its reduced production of the toxin deoxynivalenol in host tissue. All of these defects were restored by genetic complementation of the mutant with the parental FgFim gene. Quantitative real-time polymerase chain reaction (PCR) assays showed that the basal expression of three Cyp51 genes, which encode sterol 14α-demethylase, was significantly lower in the mutant than in the parental strain. The results of this study indicate that FgFim plays a critical role in the regulation of resistance to JS399-19 and in various cellular processes in F. graminearum.