Verticillium dahliae VdTHI20, Involved in Pyrimidine Biosynthesis, Is Required for DNA Repair Functions and Pathogenicity

Int J Mol Sci. 2020 Feb 18;21(4):1378. doi: 10.3390/ijms21041378.

Abstract

Verticillium dahliae (V. dahliae) infects roots and colonizes the vascular vessels of host plants, significantly reducing the economic yield of cotton and other crops. In this study, the protein VdTHI20, which is involved in the thiamine biosynthesis pathway, was characterized by knocking out the corresponding VdTHI20 gene in V. dahliae via Agrobacterium tumefaciens-mediated transformation (ATMT). The deletion of VdTHI20 resulted in several phenotypic defects in vegetative growth and conidiation and in impaired virulence in tobacco seedlings. We show that VdTHI20 increases the tolerance of V. dahliae to UV damage. The impaired vegetative growth of ΔVdTHI20 mutant strains was restored by complementation with a functional copy of the VdTHI20 gene or by supplementation with additional thiamine. Furthermore, the root infection and colonization of the ΔVdTHI20 mutant strains were suppressed, as indicated by green fluorescent protein (GFP)-labelling under microscope observation. When the RNAi constructs of VdTHI20 were used to transform Nicotiana benthamiana, the transgenic lines expressing dsVdTHI20 showed elevated resistance to V. dahliae. Together, these results suggest that VdTHI20 plays a significant role in the pathogenicity of V. dahliae. In addition, the pathogenesis-related gene VdTHI20 exhibits potential for controlling V. dahliae in important crops.

Keywords: THI20; Verticillium dahliae; pathogenicity; thiamine biosynthesis.

MeSH terms

  • Biosynthetic Pathways* / drug effects
  • Biosynthetic Pathways* / genetics
  • DNA Repair* / drug effects
  • Fluorescence
  • Fungal Proteins / genetics
  • Fungal Proteins / metabolism*
  • Gene Deletion
  • Gene Expression Regulation, Fungal / drug effects
  • Gene Expression Regulation, Fungal / radiation effects
  • Genetic Complementation Test
  • Green Fluorescent Proteins / metabolism
  • Mutation / genetics
  • Mycelium / drug effects
  • Mycelium / growth & development
  • Mycelium / metabolism
  • Nicotiana / microbiology
  • Plant Roots / drug effects
  • Plant Roots / microbiology
  • Plants, Genetically Modified
  • Pyrimidines / biosynthesis*
  • Thiamine / pharmacology
  • Ultraviolet Rays
  • Verticillium / drug effects
  • Verticillium / growth & development
  • Verticillium / metabolism*
  • Verticillium / pathogenicity*
  • Virulence / drug effects
  • Virulence / genetics
  • Virulence / radiation effects

Substances

  • Fungal Proteins
  • Pyrimidines
  • Green Fluorescent Proteins
  • pyrimidine
  • Thiamine