摘要
目的：建立一套根癌农杆菌转化枳壳的有效方法，为枳壳新品种选育、引入外源基因奠定基础。方法：以枳壳实生苗的上胚轴为材料，农杆菌菌株为ＥＨＡ１０１，含质粒载体ＰＧＡ４８２ＧＧ，质粒上插入了ＧＵＳ，ＮＰＴⅡ基因。结果：２０ｄ苗龄的外植体转化再生率较高；其感染前先进行２ｄ预培养，对转化有一定促进作用；外植体与农杆菌共培养时间以２～３ｄ为宜；乙酰丁香酮能较大提高转化效率，转化再生频率达２７．５％；外植体与农杆菌共培养后延迟２ｄ选择，抗性芽发生率有所提高。结论：通过探讨根癌农杆菌对枳壳遗传转化的影响因素，获得较高的转化再生频率。
Objective:Establishing an effective system for the Agrobacterium mediated genetic transformation of Poncirus trifoliata,so as to lay a foundation for improving breeds and introducing foreign genes.Method:The explants used for the genetic transformation were the epicotyls from Poncirus trifoliata.The Agrobacterium tumefaciens strain was EHA 101 ,containing vector plasmid PGA482GG.The GUS gene and NPTⅡ gene were introduced into the transformation plasmid.Results:20 day epicotyls were suitable for transformation.2 day precultivation enhanced shoot regeneration.Shoot regeneration frequency was high when the cocultivation lasted 2～3 days.With the presence of acetosyringone during cocultivation, the efficiency of transformation could be greatly enhanced,and the frequency of shoot regeneration rose as high as 27.5%.After the explants were cocultivated with Agrobacterium,2 day delayed selection could stimulate shoot regeneration.Conclusion:An effective genefic transformation system has been established for Poncirus trifoliata on the basis of factors affecting Agrobacterium mediated genetic transformation.
出处
《中国中药杂志》
CAS
CSCD
北大核心
1999年第3期140-142,共3页
China Journal of Chinese Materia Medica
基金
香港Croucher基金
关键词
根癌农杆菌
枳壳
遗传转化
Agrobacterium tumefaciens
Poncirus trifoliata
genetic transformation
