摘要
利用带有内含子gus基因的瞬时表达,研究影响根癌农杆菌介导"鬼露甘"草莓遗传转化的若干因素。结果表明:草莓叶片外植体对卡那霉素较为敏感,适宜的筛选浓度为20mg/L,可明显抑制非转化组织的生长;300mg/L的羧吩青霉素可有效抑菌,对外植体生长影响也较小;预培养2-4d有利于转化;共培养2-3d有利于提高转化频率并避免了农杆菌的过度生长;OD600nm值为0.4的菌液侵染10min效果最佳。再生植株经PCR检测初步证明gus基因已成功整合到草莓基因组中。
Several factors affecting Agrobacterium tumefaciens-mediated gene transformation of strawberry were studied by using the transient expression of gus gene with introns as criteria. The results showed that： the explants of strawberry leaves were rather sensitive to kanamycin, and the corresponding concentration of 20 mg/L could substantially inhibit the growth of non- transformed tisse and be used to screen the leaves of strawberry after transformation. Timentin at the concentration of 300 mg/L could be used to eliminate Agrobacterium tumefaciens and had minor impact on the growth of the explants. The precuhure time of 2 - 4 days was beneficial to improve transformation frequency. Co-culture for 2 - 3 days not only had higher gus transient expression but also avoid the over-growth of agrobacterium. The bacterial concentration of OD600mm 0.4 was the best inoculum density and 10 minutes was the best inoculation time. The PCR analysis proved that the target gene gus was successfully integrated into the genome of strawberry.
出处
《核农学报》
CAS
CSCD
北大核心
2008年第1期36-40,35,共6页
Journal of Nuclear Agricultural Sciences
关键词
根癌农杆菌
草莓
遗传转化
Agrobacterium tumefaciens
strawberry
genetic transformation
