摘要
植物基因转化的成功依赖于一个良好的转化系统,能有效地将外源基因导入受体细胞,并得以表达。通过农杆菌介导的马铃薯遗传转化体系主要受基因型、预培养、菌液浓度及侵染时间、共培养等因素的影响,由于转化受体的异质性,有必要根据实际情况进行验证和改进,以获得最适转化条件。本研究以马铃薯无菌苗的叶片、茎段为外植体,通过农杆菌介导法,将抗马铃薯X病毒和Y病毒的RNA干扰型基因结构转入马铃薯。通过研究外植体预培养、菌液浓度及侵染时间、共培养等不同转化条件及影响因素对马铃薯遗传转化的影响,建立一种高效的马铃薯遗传转化体系,试验得出最佳转化条件为外植体预培养2 d,然后用OD600=0.5的农杆菌液侵染10 min,共培养2 d。本研究为下一步的对PVX和PVY双抗的马铃薯无标记转基因研究提供技术参考。
The success of plant gene transformation depends on a good transformation system,which can effectively import the exogenous gene into the receptor cell and express it.Genetic transformation system mediated by Agrobacterium mainly affected by potato genotypes,pre-culture,concentration of bacterium,infection time,co-culture and other factors.It is necessary to verify and improve the actual situation to obtain the optimal transformation conditions because of the transformation receptor heterogeneity.In this study,the sterile potato leaves and stem segments were explants.RNA interfering genetic structure which anti-potato X virus and Y virus were transferred into the potato by Agrobacterium mediated method.To establish an efficient potato genetic transformation system by studying the explant pre-culture,concentration of bacterium,infection time,co-culture and the other influence factors on the genetic transformation on potato.The optimum transformation conditions were 2 days pre-culture of the explants,10 min infected with the Agrobacterium with OD600=0.5 and 2 days co-cultivated.And it provided technical reference for the unmarked transgenic study of PVX and PVY double-resistant potato.
作者
张西英
张爱萍
刘江娜
Zhang Xiying;Zhang Aiping;Liu Jiangna(Institute of Agriculture Science,Agriculture Division 6th,Xinjiang Produsction and Construction Corps,Wujiaqu,830000)
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2019年第7期3174-3179,共6页
Genomics and Applied Biology
基金
新建生产建设兵团第六师五家渠科技计划项目马铃薯抗病毒无标记转基因研究(1506)资助
关键词
马铃薯
转基因
转化体系
Potatoes
Genetically modified
Conversion system
