禾本科镰刀菌效应蛋白酶 FgTPP1 可抑制免疫反应并促进镰刀菌头疫病的发生

Martin Darino, Namrata Jaiswal, Reynaldi Darma, Erika Kroll, Martin Urban, Youhuang Xiang, Hye-Seon Kim, Ariana Myers, Steven Scofield, Roger W Innes, Kim E. Hammond-Kosack, Matthew Helm
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摘要

大多数植物病原体都会分泌效应蛋白来规避宿主的免疫反应,从而提高病原体的毒力。禾本科镰刀菌(Fusarium graminearum)就是这样一种病原体,它会导致小麦和大麦发生镰刀菌头疫病(FHB)。转录组分析表明,禾谷镰刀菌在感染过程的早期阶段会表达许多候选效应蛋白,其中一些被注释为蛋白酶。然而,这些蛋白酶对毒力的贡献仍不十分明确。在此,我们描述了禾谷镰孢内肽酶 FgTPP1(FGSG_11164)的特性,它在小麦穗感染过程中高度上调,并从真菌细胞中分泌。为了阐明 FgTPP1 在禾谷镰孢毒力中的潜在作用,我们生成了 FgTPP1 缺失突变体(ΔFgtpp1)并进行了 FHB 感染试验。被禾谷镰孢野生型感染的完全漂白穗的数量达到总感染穗的 50%,而被ΔFgtpp1 突变体感染的完全漂白穗的数量为 25%,这表明 FgTPP1 对真菌的毒力有贡献。绿色荧光蛋白(GFP)标记的 FgTPP1 的瞬时表达显示,FgTPP1 部分定位于叶绿体,在植物体内表达时,可减轻几丁质介导的丝裂原活化蛋白激酶(MAPK)信号激活、活性氧产生以及自身活性抗病蛋白诱导的细胞死亡。值得注意的是,FgTPP1 蛋白在子囊菌门中是保守的,使其成为子囊菌植物病原体的核心效应物。这些特性使 FgTPP1 成为诱饵底物工程的理想候选蛋白,其目标是工程化抵抗 FHB 以及由子囊真菌引起的其他作物病害。
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The Fusarium graminearum effector protease FgTPP1 suppresses immune responses and facilitates Fusarium Head Blight Disease
Most plant pathogens secrete effector proteins to circumvent host immune responses, thereby promoting pathogen virulence. One such pathogen is the fungus Fusarium graminearum, which causes Fusarium Head Blight (FHB) disease on wheat and barley. Transcriptomic analyses revealed that F. graminearum expresses many candidate effector proteins during early phases of the infection process, some of which are annotated as proteases. However, the contributions of these proteases to virulence remains poorly defined. Here, we characterize a F. graminearum endopeptidase, FgTPP1 (FGSG_11164), that is highly upregulated during wheat spikelet infection and is secreted from fungal cells. To elucidate the potential role of FgTPP1 in F. graminearum virulence, we generated FgTPP1 deletion mutants (ΔFgtpp1) and performed FHB infection assays. While the number of completely bleached spikes infected by F. graminearum wild-type reached 50% of total infected spikes, the number of fully bleached spikes infected by ΔFgtpp1 mutants was 25%, suggesting FgTPP1 contributes to fungal virulence. Transient expression of green fluorescent protein (GFP)-tagged FgTPP1 revealed that FgTPP1 localizes, in part, to chloroplasts and attenuates chitin-mediated activation of mitogen-activated protein kinase (MAPK) signaling, reactive oxygen species production, and cell death induced by an autoactive disease resistance protein when expressed in planta. Notably, the FgTPP1 protein is conserved across the Ascomycota phylum, making it a core effector among ascomycete plant pathogens. These properties make FgTPP1 an ideal candidate for decoy substrate engineering, with the goal of engineering resistance to FHB, and likely other crop diseases caused by ascomycete fungi.
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