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棉花学报 ›› 2015, Vol. 27 ›› Issue (4): 346-353.doi: 10.11963/issn.1002-7807.201504007

• 研究与进展 • 上一篇    下一篇

棉花黄萎病菌VdSge1基因的敲除与功能分析

高云,孟佩,张婷,黄家风*, 高峰*   

  1. 石河子大学农学院/新疆绿洲农业病虫害治理与植保资源利用自治区高校重点实验室,新疆 石河子 832003
  • 收稿日期:2015-04-13 出版日期:2015-07-15 发布日期:2015-07-15
  • 通讯作者: 黄家风,jiafeng_ huang@163.com;*并列通信作者,高峰,gf_agr@shzu.edu.cn
  • 作者简介:高云(1988―),女,硕士生,547452811@qq.com
  • 基金资助:
    国家自然科学基金(31160351,31160350);新疆生产建设兵团农作物病害安全防控创新团队(2101802)

Knockout and Functional Analysis of VdSge1 in Verticillium dahliae

Gao Yun, Meng Pei, Zhang Ting, Huang Jiafeng*, Gao Feng*   

  1. College of Agriculture, Shihezi University /Key Laboratory at Universities of Xinjiang Uygur Autonomous Region for Oasis Agricultural Pest Management and Plant Protection Resource Utilization, Shihezi, Xinjiang 832003, China
  • Received:2015-04-13 Online:2015-07-15 Published:2015-07-15

摘要: 为了明确棉花黄萎病菌大丽轮枝菌(Verticillium dahliae Kleb.)的VdSge1基因功能,本研究利用基因同源重组原理,通过构建敲除载体,对棉花黄萎病菌落叶型强致病力菌株V592的VdSge1基因进行了敲除,获得了4个目标基因敲除的突变体;以野生型菌株V592为对照,通过对敲除突变体生物学性状和致病性测定,结果表明,突变体的菌落生长速度、产孢量明显高于野生型,并且丧失对棉花的致病性。利用qRT-PCR对其他基因在突变体中的表达情况进行分析,结果表明,VdSge1基因敲除后,VMK1、VGBVdGARP1的表达量随之下降,而VDH1和PevD1的表达却明显上升。由此说明,VdSge1基因与大丽轮枝菌的菌落生长速度、产孢量及致病性密切相关,并且可以影响其他一些基因的表达。

关键词: 棉花; 大丽轮枝菌; VdSge1基因; 基因敲除; 基因功能

Abstract: Verticillium dahliae Kleb. is a phytopathogenic fungus and causative agent of cotton wilt disease. To characterize the V. dahliae Kleb. gene VdSge1, a targeted gene replacement strategy based on homologous recombination was adopted to generate VdSge1 deletion mutant strains from the virulent defoliating V. dahliae isolate V592 by Agrobacterium tumefaciens-mediated transformation. Four VdSge1 deletion mutants were selected for biological function and pathogenicity assays. All VdSge1 deletion mutants displayed greater radial growth and conidia production, as well as a loss in pathogenicity on cotton plants when compared to wild type V592. We also analyzed the expression of series of well-known functional genes in both V592 strain and VdSge1 deletion mutants by quantitative reverse-transcription polymerase chain reaction. The results showed that VdSge1 affects the transcription of five genes; in compared to V592 strain, VMK1, VGB and VdGARP1 were significantly up-regulated in VdSge1 deletion mutants, whereas VDH1 and PevD1 were strongly down-regulated in VdSge1 deletion mutants. In conclusion, our data suggest that V. dahliae VdSge1 plays a role in fungal development and pathogenicity, in addition to affecting the expression of other genes in this pathogenic fungus.

Key words: cotton; Verticillium dahlia; VdSge1 gene; gene knockout; gene function

中图分类号: 
  • S435.621