Construction of T-DNA Insertional Mutant Library for V erticillium dahliae and Identification of Pathogenicity Defective M utants
GU Su-jing
WANG Min
SANG Xi
ZHAO Jing
LI Hong-lian
Abstract：The fungus Verticillium dahliae causes cotton Verticillium wilt ,one of the most devas-tating diseases of cotton ,however ,the molecular basis of pathogenicity of V .dahliae is unclear .In this study ,Agrobacterium tumefaciens-mediated transformation(ATMT)was applied for inser-tional mutagenesis of V .dahliae FGH2 ,a highly virulent and defoliating strain .The collection of 1 200 T-DNA random insertion transformants of V .dahliae was generated ,using the optimized cocultivating conditions of V .dahliae conidia and A grobacterium(1∶250 ratio )for 48 h ,with the AS concentration of 400 μmol/L and the nitrocellulose filter as the filter type in the co-cultiva-tion ,resulting in 612 .5 transformants per 106 conidia .T hree pathogenicity defective V .dahliae mutants were obtained through testing the pathogenicity of 300 T-DNA transformants on the cot-ton variety Yinshan 1 seedling by dipping roots with spore suspension ,including one T-DNA in-sertion mutant losing the ability for forming microsclerotia and one T-DNA insertion mutant de-fective of microsclerotia development capability .The T-DNA insertional mutagenesis library of V . dahliae lays a foundation for researching molecular mechanisms of pathogenicity and microsclero-tia development of V .dahliae .
Keywords：cotton Verticillium wiltVerticillium dahliaeAgrobacterium tumefaciens-mediated transformationT-DNA insertional mutagenesispathogenicity
Publication Date：2014-01-01
Online Publishing Date：2026-09-12(First online date of this platform, not the publication date of the document)
Pages：6( 69-73,83 )