Establishing of the Transformation System of Fusarium oxysporum f. sp. melongenae by Agrobacterium tumefaciens-mediated Transformation (ATMT) and Constructing of GFP Labeled Strain
YAN Yan-Qin, HU Tian-Hua, WANG Wu-Hong, HU Hai-Jiao, WEI Qing-Zhen, WANG Jing-Lei, BAO Chong-Lai*
Institute of Vegetable, Zhejiang Academy of Agricultural Science, Hangzhou 310000, China
Abstract：Fusarium wilt caused by Fusarium oxysporum f. sp. melongenae seriously restricts the production of eggplant (Solanum melongena). However, its pathogenic mechanism is still unclear. In this study, a transformation system of F. oxysporum f. sp. melongenae was established and optimized, in which GFP expression vector pCAMsgfp was transformed into conidia using , ATMT (Agrobacterium tumefaciens-mediated transformation) method. The results showed that a satisfactory transformation with an efficiency of 319±10.21 transformants per 1×106 spores could be achieved in a system of 1×106 spores per milliliter of F. oxysporum f. sp. melongenae spore suspension which were co-cultured with Agrobacterium tumefaciens suspension at a concentration of OD600 0.5 under the co-culture medium containing 200 μmol/mL acetosyringone at 25~28 ℃ for 48 h. PCR identification of the GFP gene and fluorescence observation showed that GFP gene could be expressed normally. In this study, the genetic transformation system of F. oxysporum f.sp. melongenae mediated by ATMT was established and optimized, and the GFP-labeled strain was successfully obtained, which could provide powerful technic for further explore the infection process and the pathogenic mechanism of F. oxysporum f. sp. melongenae.
严亚琴, 胡天华, 王五宏, 胡海娇, 魏庆镇, 汪精磊, 包崇来. 基于农杆菌介导的茄子枯萎病菌遗传转化体系建立及GFP标记菌株的构建[J]. 农业生物技术学报, 2023, 31(9): 1980-1988.
YAN Yan-Qin, HU Tian-Hua, WANG Wu-Hong, HU Hai-Jiao, WEI Qing-Zhen, WANG Jing-Lei, BAO Chong-Lai. Establishing of the Transformation System of Fusarium oxysporum f. sp. melongenae by Agrobacterium tumefaciens-mediated Transformation (ATMT) and Constructing of GFP Labeled Strain. 农业生物技术学报, 2023, 31(9): 1980-1988.
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