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农杆菌介导樱桃干腐病菌的遗传转化

DOI: 10.3969/j.issn.1000-7091.2013.04.040, PP. 218-222

Keywords: 樱桃干腐病菌,遗传转化,转化效率,转化子鉴定

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Abstract:

以携带潮霉素磷酸转移酶基因的pBIG3C为转化载体,根癌农杆菌EHA105为转化介体,对樱桃干腐病菌LXS230101分生孢子进行转化。结果表明,樱桃干腐病菌的最优转化体系为:α型分生孢子浓度为106个/mL,培养基中添加200μmol/mL乙酰丁香酮(AS),共培养温度和时间分别为25℃和72h,其转化效率为1×106个α型分生孢子产生686个转化子。随机选取转化子进行PCR和Southernblot鉴定,发现T-DNA已整合进樱桃干腐病菌基因组中;转化子在不含潮霉素的PDA培养基中连续培养5代后,仍表现出对潮霉素的抗性,表明外源基因能在樱桃干腐病菌中稳定遗传。

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