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期 刊 ：中国中药杂志 1999年 24卷 03期 页码：140.

关键词：根癌农杆菌；枳壳；遗传转化；

Keywords:Agrobacterium tumefaciens, Poncirus trifoliata, genetic transformation,

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，建立了有效的转化再生系统。

Abstract: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.

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