High efficient gene knockout in Verticillium dahliae

Abstract:

Abstract:［Objective］We developed an efficient method of gene knockout in Verticillium dahliae，an important soil-borne fungal pathogen that causes cotton vascular wilt diseases． ［Methods］By using fusion PCR，we constructed gene knockout vectors． By using Agrobacterium tumefaciens-mediated transformation and applying a herpes simplex virus thymidine kinase (HSVtk) gene in T-DNA as a conditional lethal gene to counter-select against ectopic transformants，we developed an efficient method to select gene knockout transformants． ［Results］Gene knockout frequency for ADE4 and ChsV was 87% and 44% ，respectively． ［Conclusion］We developed an efficient tool for gene knockout in Verticillium dahliae，which would help clarify the infection mechanism of this fungal pathogen．