Journal of Integrative Agriculture ›› 2025, Vol. 24 ›› Issue (10): 3941-3952.DOI: 10.1016/j.jia.2024.01.029

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FoVEL1 和 FoLAE1基因影响西瓜枯萎病菌分生孢子、致病力和次生代谢

  

  • 收稿日期:2023-09-04 修回日期:2024-01-20 接受日期:2023-12-01 出版日期:2025-10-20 发布日期:2025-09-24

Involvement of FoVEL1 and FoLAE1 in conidiation, virulence and secondary metabolism of Fusarium oxysporum f. sp. niveum

Yang Sun1, 2*, Xuhuan Zhang1*, Zhenqin Chai1, Yuying Li1, Zheng Ren3, Miaomiao Wang1, Zhiqing Ma1, Yong Wang1#, Juntao Feng1#   

  1. 1 College of Plant Protection, Northwest A&F University, Yangling 712100, China

    2 College of Plant Protection, Anhui Agricultural University, Hefei 230036, China

    3 College of Language and Culture, Northwest A&F University, Yangling 712100, China

  • Received:2023-09-04 Revised:2024-01-20 Accepted:2023-12-01 Online:2025-10-20 Published:2025-09-24
  • About author:Yang Sun, E-mail: Sunyang@ahau.edu.cn; Xuhuan Zhang, E-mail: zhangxuhuan01@163.com; #Correspondence Yong Wang, E-mail: wy2010102163@163.com; Juntao Feng, E-mail: jtfeng@126.com * These authors contributed equally to this study.
  • Supported by:
    This investigation was supported by the National Natural Science Foundation of China (32072461) and the Open Foundation of Shaanxi Key Laboratory of Plant Nematology, China (2021-SKL-01). 

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摘要：

[目的] 研究Velvet蛋白家族FoVEL1基因和甲基转移酶FoLAE1基因在西瓜枯萎病菌中的功能。[方法] 以西瓜枯萎病菌为研究对象，构建FoVEL1和 FoLAE1基因的敲除突变体（∆fovel1，∆folae1）和回补突变体（∆fovel1-C，∆folae1-C）；分别测定敏感菌株及各突变体的菌丝生长速率和分生孢子萌发情况；使用电导率计测定敏感菌株及各突变体分别在0、50、100、150、200和250分钟后的电导率，以反映细胞膜通透性；测定敏感菌株及各突变体对不同胁迫条件（光，NaCl，SDS，H2O2）的敏感性；将敏感菌株及各突变体培养7天后，用UV-5100紫外分光光度计测定镰刀菌酸含量；盆栽试验测定敏感菌株及各突变体的致病力；将敏感菌株及各突变体培养7天后，提取总RNA反转录为cDNA，利用实时荧光定量分析各突变体中与致病性相关的3个基因（FOW1、FMK1、MPK1）及比卡菌红素主要调控基因（BIK1，BIK2和BIK3）表达水平；利用酵母双杂测定FoVel1和 FoLae1的互作关系。[结果] 与野生型菌株相比，∆fovel1 和 ∆folae1 突变体表现出不同的菌丝体表型，色素沉着显著减少，分生孢子数量显著减少。qRT-PCR结果表明与野生型菌株相比，∆fovel1和∆folae1突变体中BIK1、BIK2和BIK3表达水平显著降低。电导率实验和胁迫实验表明，∆fovel1突变体的细胞膜通透性显著提高，对渗透压、细胞壁抑制剂和氧化压力等胁迫条件的敏感性略有提高。镰刀菌酸是西瓜枯萎病菌产生的主要毒素之一，∆fovel1和∆folae1突变体的镰刀菌酸含量明显减少，侵染能力显著降低。qRT-PCR结果表明，∆fovel1和∆folae1突变体中与致病性相关的三个基因（FOW1，FMK1，MPK1）表达水平显著低于野生型菌株。酵母双杂结果表明，Velvet蛋白FoVel1可与位于细胞核的甲基转移酶FoLae1相互作用。 [结论] 西瓜枯萎病菌中，Velvet蛋白家族FoVEL1基因和甲基转移酶FoLAE1基因在分生孢子产生、致病力和调节次生代谢中起关键作用。

Abstract:

The velvet protein family serves as a crucial factor in coordinating development and secondary metabolism in numerous pathogenic fungi.  However, no previous research has examined the function of the velvet protein family in Fusarium oxysporum f. sp. niveum (FON), a pathogen causing a highly destructive disease in watermelon.  In this study, ∆fovel1 and ∆folae1 deletion mutants and ∆fovel1-C and ∆folae1-C corresponding complementation mutants of FON were validated.  Additionally, the phenotypic, biochemical, and virulence effects of the deletion mutants were investigated.  Compared to the wild-type strains, the ∆fovel1 and ∆folae1 mutants exhibited altered mycelial phenotype, reduced conidiation, and decreased production of bikaverin and fusaric acid.  Furthermore, their virulence on watermelon plant roots significantly decreased.  All these alterations in mutants were restored in corresponding complementation strains.  Notably, yeast two-hybrid results demonstrated an interaction between FoVel1 and FoLae1.  This study reveals that FoVEL1 and FoLAE1 play essential roles in secondary metabolism, conidiation, and virulence in FON.  These findings enhance our understanding of the genetic and functional roles of VEL1 and LAE1 in pathogenic fungi.

Key words: velvet complex ,  Fusarium oxysporum f. sp. Niveum ,  fusaric acid ,  virulence , protein interaction