CAJ | 학술논문

大丽轮枝菌（Verticillium dahliae）侵染引起的棉花黄萎病是棉花生产上的重要病害之一，而其致病的分子机制尚不清楚，为此，以强致病力落叶型黄萎病菌FGH2为材料，利用农杆菌介导转化技术构建棉花黄萎病菌T-DNA插入突变体库并进行致病缺陷突变体的筛选。当农杆菌与黄萎病菌分生孢子浓度比为250∶1、承载介质为NC膜、乙酰丁香酮浓度400μm o l/L、共培养时间48 h时，转化效率高达每106个分生孢子产生612．5个转化子。通过采用优化后的转化体系，构建了含1200个T-DNA插入转化子的棉花黄萎病菌突变体库。利用苗期分生孢子蘸根法测定300个转化子在棉花幼苗上的致病力，获得3个致病力显著降低的突变体181、546、1009，其中546的微菌核形成能力丧失，1009的微菌核形成能力明显下降。黄萎病菌T-DNA插入突变体库的构建，为棉花黄萎病菌致病及微菌核形成机制的研究奠定了基础。
대려륜지균（Verticillium dahliae）침염인기적면화황위병시면화생산상적중요병해지일，이기치병적분자궤제상불청초，위차，이강치병력락협형황위병균FGH2위재료，이용농간균개도전화기술구건면화황위병균T-DNA삽입돌변체고병진행치병결함돌변체적사선。당농간균여황위병균분생포자농도비위250∶1、승재개질위NC막、을선정향동농도400μm o l/L、공배양시간48 h시，전화효솔고체매106개분생포자산생612．5개전화자。통과채용우화후적전화체계，구건료함1200개T-DNA삽입전화자적면화황위병균돌변체고。이용묘기분생포자잠근법측정300개전화자재면화유묘상적치병력，획득3개치병력현저강저적돌변체181、546、1009，기중546적미균핵형성능력상실，1009적미균핵형성능력명현하강。황위병균T-DNA삽입돌변체고적구건，위면화황위병균치병급미균핵형성궤제적연구전정료기출。
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 .