摘要
目的 :采用农杆菌介导法 ,将能抑制病毒侵染的外壳蛋白基因转入枳壳 ,为通过基因转化进行枳壳的抗病育种提供依据和方法。方法 :以枳壳实生苗上胚轴为材料 ,农杆菌菌株为EHA10 1,含有质粒载体 pGA482GG。质粒上插入了 35s启动子驱动的外源目的基因———柑桔衰退病病毒外壳蛋白基因 (CTV cp)及GUS ,NPTⅡ基因。结果 :枳壳转化试验中 ,抗菌素采用头孢霉素较好 ,浓度为 30 0mg·L-1;以卡那霉素作为选择试剂 ,浓度为 5 0mg·L-1;转化获得的抗性植株 ,GUS检测结果 ,70 .0 %的植株呈阳性反应 ;Southern杂交分析 (Southernblot)鉴定证明外壳蛋白基因已整合到枳壳植株的核基因组中。结论 :通过农杆菌介导的转化法 ,成功地获得了转外壳蛋白基因的枳壳植株。
Objective:To lay a foundation for the resistant breding, the anti virus CTV cp gene was transformed into the epicotyles mediated by Agrobacterium tumefaciens in Poncirus trifoliata . Method:The explants used for the genetic transformation were the epicotyls from P.trifoliata. The Agrobacterum tumefaciens strain was EHA 101 containing vector plasmid pGA482GG.The coat protein gene (CTV cp gene),GUS gene and NPT II gene were introduced into the transformation plasmid.Results:Ceftaxime used as antibiotics was better than carbenicillin. The concentration was 300 mg·L -1 ;The selection pressure for kanamycin was 50 mg·L -1 ;70.0% of the resistant plants were GUS positive; extra gene was proved to be in P.trifoliata plant by southern blot examination. Conclusion:An effective genetic transformation mediated by Agrobacterium tumefacines, which harbours a CTV cp gene, has been developed in P.trifoliata. Transgenic CTV cp plants were obtained. [
出处
《中国中药杂志》
CAS
CSCD
北大核心
2001年第1期21-23,共3页
China Journal of Chinese Materia Medica
基金
国家自然科学基金!项目 (396 70 476 )
关键词
枳壳
外壳蛋白基因
植物转化
中药
P.trifoliata
coat protein gene
plant transformat8
