农杆菌介导的稻瘟病菌遗传转化体系的建立及优化  

Establishment and optimization of Agrobacterium mediated genetic transformation system for Magnaporthe oryzae

作　　者：李爽 张淑梅[1] 田缘 潘钰[1,3] 刘伟[1] 闫更轩[1] LI Shuang;ZHANG Shu-mei;TIAN Yuan;PAN Yu;LIU Wei;YAN Geng-xuan(Institute of Microbiology,Heilongjiang Academy of Sciences,Harbin150010,China;College of Food Science,Northeast Agricultural University,Harbin150030,China;School of Life Sciences,Heilongjiang University,Harbin150006,China)

机构地区：[1]黑龙江省科学院微生物研究所,哈尔滨150010 [2]东北农业大学食品学院,哈尔滨150030 [3]黑龙江大学生命科学学院,哈尔滨150006

出　　处：《湖北农业科学》2025年第3期182-189,共8页Hubei Agricultural Sciences

基　　金：省属科研院所科研业务费项目(CZKYF2021-2-C031)。

摘　　要：以稻瘟病菌(Magnaporthe oryzae)为试验材料,通过PCR获得GFP基因,构建pBarg-GFP-BARAmp重组载体,使用农杆菌转化法获得能够稳定遗传的含GFP基因的稻瘟病菌菌株,采用单因素与响应面法优化转化条件。稻瘟病菌遗传转化体系的最佳转化条件:OD_(600 nm)为0.38(农杆菌菌液浓度)、共培养温度为28.2℃、共培养时间为2.12 d,转化效率为280.00×10~(-5)。PCR鉴定和荧光显微镜观察发现,转化的稻瘟病菌GFP基因能够正常表达,和野生型稻瘟病菌的致病性无显著差异。Magnaporthe oryzae was used as the experimental material.The GFP gene was obtained through PCR,and a pBarg-GFP-BAR-Amp recombinant vector was constructed.The Magnaporthe oryzae strain containing the GFP gene with stable inheritance was obtained using Agrobacterium-mediated transformation.The transformation conditions were optimized using single factor and response surface methodology.The optimal transformation conditions for the Magnaporthe oryzae genetic transformation system were an OD600 nm of 0.38(Agrobacterium concentration),a co-culture temperature of 28.2℃,a co-culture time of 2.12 days,and a transformation efficiency of 280.00×10-5.PCR identification and fluorescence microscopy observation revealed that the GFP gene of the transformant Magnaporthe oryzae could be expressed normally,with no significant difference in pathogenicity compared to the wild-type Magnaporthe oryzae.

关 键 词：稻瘟病菌(Magnaporthe oryzae) 农杆菌介导 遗传转化 GFP基因 建立 优化 

分 类 号：S432.4[农业科学—植物病理学]