基于ATMT的茄子黄萎病菌遗传转化与转化子特性分析OA

Taking the highly pathogenic strain EP-3 of eggplant Verticillium wilt fungus as the test material,an Agrobacterium tumefaciens-mediated genetic transformation(ATMT)system was employed to systematically optimize key transformation parameters,including concentrations of A.tume faciens and acetosyringone(AS),as well as co-culture temperature and time,to establish optimal transformation conditions.The genetic expression stability of the obtained GFP-labeled transformants was assessed by analyzing characteristics such as growth rate,sporulation capacity,crude toxin secretion,and in order to pathogenicity,to establish and optimize a genetic transformation system for eggplant Verticillium wilt fungus and obtain green fluorescent protein(GFP)-labeled strains,and in order to provide reference for subsequent studies on the infection and colonization process of this pathogen in eggplants and its pathogenic mechanisms.The results showed that the optimal transformation conditions were as follows,A.tumefaciens concentration at OD600=0.6,conidial concentration of 1×105 conidia·mL-1,acetosyringone(AS)concentration of 200 μmol·L-1,and co-culture at 25 ℃ for 48 hours.Through hygromycin resistance screening,PCR,and fluorescence microscopy,it was confirmed that the GFP gene had been successfully integrated into the genome of strain EP-3 and was stably expressed.Analysis of transformant characteristics indicated that the obtained transformants exhibited stable core biological traits,with no significant differences compared to the wild-type strain.These findings further demonstrate that an efficient genetic transformation system for eggplant Verticillium wilt fungus has been successfully established in this study.