根癌农杆菌介导的玉米大斑病菌转化条件的优化

Optimization of factors affecting genetic transformation of Setosphaeria turcica via Agrobacterium tumefaciens

为构建大规模的玉米大斑病菌ATMT突变体库,本研究对根癌农杆菌介导的玉米大斑病菌的转化条件进行了优化,包括共培养温度与时间、孢子萌发时间、转化受体材料、共培养介质等。结果表明:头孢噻肟钠和羧苄青霉素浓度为200/200μg/mL时可有效抑制根癌农杆菌的生长;当玉米大斑病菌分生孢子萌发24 h后,在20℃条件下与根癌农杆菌共培养48 h,共培养介质为微孔滤膜时转化效果最佳,转化效率可达到90～110个转化子/105个孢子。

In order to construct a large-scale Setosphaeria turcica ATMT mutant library,this study optimized the main factors affecting genetic transformation of S.turcica via Agrobacterium tumefaciens,including co-culturing temperature and time,spore germination time,transforming receptor materials,co-culturing media and other conditions.The results showed that the effective concentration inhibiting A.tumefaciens growth of cefotaxime and carbenicillin was 200/200 μg/mL.The most suitable conditions were as follows： germination time of S.turcica conidia was 24 h at 20 ℃,co-culturing time of the germinated conidia and bacteria was 48 h,co-culturing media was millipore filter,and the transformation frequency was up to 90-110 transformants per 105 conidia.