根癌农杆菌介导蓝猪耳转化系统的建立

 通过对影响根癌农杆菌介导蓝猪耳转化效果各种因素的研究, 建立了蓝猪耳高效稳定的遗传转化系统。研究结果表明, 农杆菌侵染叶片外植体效率最高; 外植体预培养不利于农杆菌的侵染; 乙酰丁香酮能大大提高根癌农杆菌转化蓝猪耳的效率。另外, 菌液浓度、侵染和共培养的时间及再生的途径等均对转化效率有一定的影响。叶片外植体与农杆菌共培养后, 经过抗性芽的诱导、根的再生, 70 d左右就可以获得抗性苗。抗性植株经GUS染色、PCR分析和Southern blot检测, 外源基因已经整合到蓝猪耳基因组中, 转化频率为7%～8%。

Establishment of Agrobacterium-mediated Transformation System for Torenia

An efficient and stable Agrobacterium-mediated genetic transformation system for Torenia was established through investigating the factors influencing the transformation efficiency.The results showed that leaf explants are more sensitive to Agrobacterium infection,pre-culture of explants had an adverse effect on transformation efficiency,induction of vir gene with AS greatly improved the transformation effects.In addition,the concentration of bacteria,the time of infection and co-cultivation,as well as the regeneration methods have effects on the transformation.By using the optimized conditions,hygromycin resistant plants could be obtained within 70 days from the infection of the explants with Agrobacterium.GUS staining,PCR and Southern-blot analysis confirmed that transgene were integrated into the Torenia genome with a transformation frequency of 7%-8%.