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VdOGDH 参与能量代谢,是大丽轮枝菌毒力所必需的。

VdOGDH is involved in energy metabolism and required for virulence of Verticillium dahliae.

机构信息

School of Life Science, Anhui Agricultural University, Hefei, 230036, China.

Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing, 100081, China.

出版信息

Curr Genet. 2020 Apr;66(2):345-359. doi: 10.1007/s00294-019-01025-2. Epub 2019 Aug 17.

Abstract

Verticillium dahliae, a soil-borne fungus, can invade plant vascular tissue and cause Verticillium wilt. The enzyme α-oxoglutarate dehydrogenase (OGDH), catalyzing the oxidation of α-oxoglutarate in the tricarboxylic acid cycle (TCA), is vital for energy metabolism in the fungi. Here, we identified the OGDH gene in V. dahliae (VdOGDH, VDAG_10018) and investigated its function in virulence by generating gene deletion mutants (ΔVdOGDH) and complementary mutants (ΔVdOGDH-C). When the ΔVdOGDH mutants were supplemented with different carbon sources, vegetative growth on Czapek Dox medium was significantly impaired, suggesting that VdOGDH is crucial for vegetative growth and carbon utilization. Conidia of the ΔVdOGDH mutants were atypically rounded or spherical, and hyphae were irregularly branched and lacked typical whorled branches. Mutants ΔVdOGDH-1 and ΔVdOGDH-2 were highly sensitive to HO in the medium plates and had higher intracellular ROS levels. ΔVdOGDH mutants also had elevated expression of oxidative response-related genes, indicating that VdOGDH is involved in response to oxidative stress. In addition, the disruption of VdOGDH caused a significant increase in the expression of energy metabolism-related genes VdICL, VdICDH, VdMDH, and VdPDH and melanin-related genes Vayg1, VdSCD, VdLAC, VT4HR, and VaflM in the ΔVdOGDH mutants; thus, VdOGDH is also important for energy metabolism and melanin accumulation. Cotton plants inoculated with ΔVdOGDH mutants exhibited mild leaf chlorosis and the disease index was lower compared with wild type and ΔVdOGDH-C strains. These results together show that VdOGDH involved in energy metabolism of V. dahliae, is also essential for full virulence by regulating multiple fungal developmental factors.

摘要

黄萎轮枝菌是一种土壤传播的真菌,可以侵入植物的维管束组织并引起黄萎病。α-酮戊二酸脱氢酶(OGDH)是三羧酸循环(TCA)中α-酮戊二酸氧化的关键酶,对真菌的能量代谢至关重要。在这里,我们鉴定了黄萎轮枝菌中的 OGDH 基因(VdOGDH,VDAG_10018),并通过生成基因缺失突变体(ΔVdOGDH)和互补突变体(ΔVdOGDH-C)来研究其在毒力中的作用。当 ΔVdOGDH 突变体补充不同的碳源时,在 Czapek Dox 培养基上的营养生长受到显著抑制,这表明 VdOGDH 对营养生长和碳利用至关重要。ΔVdOGDH 突变体的分生孢子呈异常圆形或球形,菌丝不规则分枝,缺乏典型的轮生分枝。突变体 ΔVdOGDH-1 和 ΔVdOGDH-2 在培养基平板上对 HO 高度敏感,细胞内 ROS 水平较高。ΔVdOGDH 突变体还上调了氧化应激相关基因的表达,表明 VdOGDH 参与了氧化应激的反应。此外,VdOGDH 的破坏导致与能量代谢相关的基因 VdICL、VdICDH、VdMDH 和 VdPDH 以及黑色素相关基因 Vayg1、VdSCD、VdLAC、VT4HR 和 VaflM 在 ΔVdOGDH 突变体中的表达显著增加;因此,VdOGDH 对于能量代谢和黑色素积累也很重要。与野生型和 ΔVdOGDH-C 菌株相比,接种 ΔVdOGDH 突变体的棉花植株表现出轻度叶片黄化,病情指数较低。这些结果表明,VdOGDH 参与了黄萎轮枝菌的能量代谢,通过调节多个真菌发育因子,对其完全毒力也是必不可少的。

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