A type 2C protein phosphatase FgPtc3 is involved in cell wall integrity, lipid metabolism, and virulence in Fusarium graminearum.
A type 2C protein phosphatase FgPtc3 is involved in cell wall integrity, lipid metabolism, and virulence in Fusarium graminearum.
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DOI：
10.1371/journal.pone.0025311
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发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Ma Z
中科院分区：
文献类型：
--
作者：
Jiang J;Yun Y;Yang Q;Shim WB;Wang Z;Ma Z
2C 型蛋白磷酸酶 (PP2C) 在调节真核生物的许多生物过程中发挥着重要作用。目前，人们对 PP2Cs 在丝状真菌中的功能知之甚少。小麦赤霉病的病原禾谷镰刀菌含有七个假定的 PP2C 基因：FgPTC1、-3、-5、-5R、-6、-7 和 -7R。为了研究这些 PP2C 的作用，我们在本研究中构建了所有 7 个 PP2C 基因的缺失突变体。 FgPTC3缺失突变体（ΔFgPtc3-8）表现出气生菌丝形成和脱氧雪腐镰刀菌烯醇（DON）产量减少，但分生孢子产量增加。该突变体在马铃薯葡萄糖琼脂平板上表现出对渗透胁迫和细胞壁损伤剂的抵抗力增强。致病性测定表明ΔFgPtc3-8不能感染开花小麦头。当 ΔFgPtc3-8 与野生型 FgPTC3 基因互补时，所有缺陷都得到恢复。此外，FgPTC3 部分挽救了酵母 PTC1 缺失突变体在各种胁迫条件下的生长缺陷。超微结构和组织化学分析表明，ΔFgPtc3-8 的分生孢子含有异常大量的大脂滴。此外，突变体比野生型祖细胞积累了更高的甘油基础水平。实时定量PCR检测显示，突变株中FgOS2、FgSLT2和FgMKK1的基础表达量显着高于野生型菌株。对 ΔFgPtc3-8 中基因表达的系列分析表明，FgPTC3 与多种代谢途径相关。与 FgPTC3 突变体相比，FgPTC1、FgPTC5、FgPTC5R、FgPTC6、FgPTC7 或 FgPTC7R 的缺失突变体在 PDA 培养基上生长或接种在麦头上时没有表现出异常表型特征。这些结果表明 FgPtc3 是关键的 PP2C，在多种细胞和生物功能中发挥着关键作用，包括细胞壁完整性、脂质和次生代谢以及禾谷镰刀菌的毒力。
Type 2C protein phosphatases (PP2Cs) play important roles in regulating many biological processes in eukaryotes. Currently, little is known about functions of PP2Cs in filamentous fungi. The causal agent of wheat head blight, Fusarium graminearum, contains seven putative PP2C genes, FgPTC1, -3, -5, -5R, -6, -7 and -7R. In order to investigate roles of these PP2Cs, we constructed deletion mutants for all seven PP2C genes in this study. The FgPTC3 deletion mutant (ΔFgPtc3-8) exhibited reduced aerial hyphae formation and deoxynivalenol (DON) production, but increased production of conidia. The mutant showed increased resistance to osmotic stress and cell wall-damaging agents on potato dextrose agar plates. Pathogencity assays showed that ΔFgPtc3-8 is unable to infect flowering wheat head. All of the defects were restored when ΔFgPtc3-8 was complemented with the wild-type FgPTC3 gene. Additionally, the FgPTC3 partially rescued growth defect of a yeast PTC1 deletion mutant under various stress conditions. Ultrastructural and histochemical analyses showed that conidia of ΔFgPtc3-8 contained an unusually high number of large lipid droplets. Furthermore, the mutant accumulated a higher basal level of glycerol than the wild-type progenitor. Quantitative real-time PCR assays showed that basal expression of FgOS2, FgSLT2 and FgMKK1 in the mutant was significantly higher than that in the wild-type strain. Serial analysis of gene expression in ΔFgPtc3-8 revealed that FgPTC3 is associated with various metabolic pathways. In contrast to the FgPTC3 mutant, the deletion mutants of FgPTC1, FgPTC5, FgPTC5R, FgPTC6, FgPTC7 or FgPTC7R did not show aberrant phenotypic features when grown on PDA medium or inoculated on wheat head. These results indicate FgPtc3 is the key PP2C that plays a critical role in a variety of cellular and biological functions, including cell wall integrity, lipid and secondary metabolisms, and virulence in F. graminearum.
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