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核农学报  2012 

农杆菌介导球孢白僵菌转化体系的建立及突变体筛选

, PP. 994-999

Keywords: 球孢白僵菌,农杆菌AGL1,T-DNA插入转化,转化效率

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Abstract:

以球孢白僵菌(Beauveriabassiana)YB-06为受体菌株,采用农杆菌介导的遗传转化方法,实现了农杆菌AGL1(pATMT1)介导的球孢白僵菌的遗传转化,并研究了转化介质和pH对转化效率的影响。结果表明在采用28℃、200μmol/L的乙酰丁香酮(AS)、农杆菌浓度OD600=0.8、孢子浓度为1×106个/ml、pH5.1~5.8时可实现T-DNA的插入转化。当转化体系pH5.3时,转化效率最高,达586个转化子/106分生孢子;不同共转化介质对转化效率影响明显,玻璃纸和硝酸纤维素膜转化效率较高。对3000个转化子生长速度和产孢量等性状分析获得46个性状特异突变体。本文结果为进一步研究球孢白僵菌的生长发育、致病机理和毒力相关基因功能奠定了基础。

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