The transcription factor FgNsf1 regulates fungal development, virulence and stress responses in Fusarium graminearum

doi:10.1016/S2095-3119(20)63339-1

摘要/Abstract

摘要：

Nsf1(Nutrient and stress factor 1)是典型的C2H2型锌指蛋白，酿酒酵母中Nsf1在非发酵碳源或者盐胁迫的条件下才会表达。进化树分析发现该基因在不同物种间比较保守，然而，Nsf1的功能在丝状真菌中研究得并不是很透彻。为了探索FgNsf1在小麦赤霉病的病原菌禾谷镰孢菌中的功能，我们构建了FgNSF1基因敲除体（ΔFgNsf1）和包含GFP标记的回复体（ΔFgNsf1-C），亚细胞定位表明FgNsf1蛋白集中于细胞核。进一步研究发现，与野生菌株PH-1和回复体ΔFgNsf1-C 相比，敲除体ΔFgNsf1的菌丝体生长速率明显减慢，分生孢子产量及萌发率显著下降，且有畸形孢子产生，子囊壳产量也显著降低。但是红色镰刀菌素和黄色镰刀菌素的产量明显增加，为了验证这一结果，我们利用实时荧光定量PCR技术检测了相关基因(AurJ, AurF, AurO, AurR2)的表达量，研究结果发现，相关基因的表达量都显著上调。此外，使用不同浓度的NaCl处理时，野生菌株PH-1中FgNSF1基因的表达量均上调，而在使用不可发酵碳源乙醇、甘油或醋酸盐作为唯一碳源时，FgNSF1基因的表达量都显著下调。另外我们发现敲除体对渗透，细胞壁，氧化和部分金属离子等胁迫因子的敏感性显著下降，只对0.2M镁离子胁迫的敏感性显著提高。药敏性实验发现，敲除体对咯菌腈和抑菌脲的抗药性明显增强，而对戊唑醇和多菌灵的敏感性显著提高。随后，我们在小麦胚芽鞘和麦穗上进行了致病力实验，结果发现ΔFgNsf1的致病力显著减弱，在产毒培养基中DON（脱氧雪腐镰刀烯醇）产量也显著下降，以及DON毒素合成相关基因TRI5和TRI6的基因表达量也显著下调。结论：FgNSF1在禾谷镰孢菌的生长发育，有性和无性生殖，应对外界胁迫，产毒和致病的过程中扮演着重要的角色。创新性：我们首次系统地报道了FgNSF1在禾谷镰孢菌中的功能。

Abstract:

Nutrient and stress factor 1 (Nsf1), a transcription factor containing the classical Cys2-His2 (C₂H₂) zinc finger motif, is expressed under non-fermentable carbon conditions and in response to salt stress in Saccharomyces cerevisiae. However, the role of Nsf1 in filamentous fungi is not well understood. In this study, the orthologue of Nsf1 was investigated in Fusarium graminearum (named FgNsf1), a causal agent of Fusarium head blight (FHB). The functions of FgNsf1 were evaluated by constructing a FgNSF1 deletion mutant, designated as ΔFgNsf1, and its functional complementation mutant ΔFgNsf1-C. Gene deletion experiments showed that the mycelial growth rate, asexual and sexual reproduction of ΔFgNsf1 were significantly reduced, but the pigment production of ΔFgNsf1 was remarkably increased compared with the PH-1 and ΔFgNsf1-C. In addition, the tolerance of ΔFgNsf1 to osmotic pressures, cell wall-damaging agents and oxidative stress increased significantly. Sensitivity tests to different fungicides revealed that ΔFgNsf1 exhibited increased sensitivity to carbendazim (MBC) and tebuconazole, and enhanced tolerance to fludioxonil and iprodione than PH-1 and ΔFgNsf1-C. The virulence of ΔFgNsf1 to wheat coleoptiles and flowering wheat heads were dramatically decreased, which was consistent with the decrease in the yield of deoxynivalenol (DON). All of these defects were restored by target gene complementation. These results indicated that FgNsf1 plays a crucial role in vegetative growth, asexual and sexual reproduction, stress responses, fungicide sensitivity, and full virulence in F. graminearum.

Key words: Fusarium graminearum , nutrient and stress factor 1 , stress responses , virulence

. [J]. Journal of Integrative Agriculture, 2021, 20(8): 2156-2169.

SHI Dong-ya, REN Wei-chao, WANG Jin, ZHANG Jie, Jane Ifunanya MBADIANYA, MAO Xue-wei, CHEN Chang-jun. The transcription factor FgNsf1 regulates fungal development, virulence and stress responses in Fusarium graminearum[J]. Journal of Integrative Agriculture, 2021, 20(8): 2156-2169.

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