烤烟栽培品种G－28、K326抗病毒遗传转化的研究  

ANTIVIRUS GENETIC TRANSFORMATION OF COMMERCUL TOBACCO CULTIVAR G-28 AND K326

作　　者：吕华飞[1] 何云昆[1] 赵淑珍[1] 王苏燕[1] 叶寅[1] 张仲凯[1] 田波[1] 

机构地区：[1]云南省农业科学院生物技术研究所,云南省农业生物技术重点实验室,中国科学院微生物研究所

出　　处：《西南农业学报》1994年第1期66-69,共4页Southwest China Journal of Agricultural Sciences

基　　金：云南省烟草公司

摘　　要：用携带植物抗病毒基因表达载体ｐＥＣ２３和ｐＲＣＰⅠ的农杆菌转化云南烤烟栽培品种Ｇ－２８和Ｋ３２６，获得了大量转基因植株，载体质粒ｐＥＣ２３上有卡那霉素抗性标记基因（ＮＰＴⅡ）和ＣａＭＶ３５ｓ启动子控制的烟草花叶病毒（ＴＭＶ）５４ＫＤ蛋白基因，ｐＲＣＰⅠ上则有卡那霉素抗性标记基因（ＮＰＴⅡ）、黄瓜花叶病毒（ＣＭＶ）外壳蛋白基因、ＣＭＶ卫星ＲＮＡ基因，品种Ｇ－２８、Ｋ３２６的叶片与携带载体质粒的农杆菌共培养后，将其按品种分别转移到含６０μｇ／ｍｌ、２０μｇ／ｍｌＫｍ的ＭＳ＋０．２ｍｇ／ｌＮＡＡ＋２．０ｍｇｌ６－ＢＡ＋０．１ｍｇ／ｌＫＴ培养基上培养筛选，２～３周后分化出芽，切下小芽分别转移到含６０μｇ／ｍｌ、２０μｇ／ｍｌＫｍ的ＭＳ＋０．２ｍｇ／ｌＮＡＡ的培养基上进一步筛选并使转化体生根，再对生根植株叶片进行抗性鉴定，结果表明，至少Ｋｍ抗性标记基因（ＮＰＴⅡ）已转入烤烟植株。Yunnan commerclal tobacco cultivar G-28 and K326 were transformed by two strains of Agrobacterium tumefaciens with the plant expression vector pEC23 and pRCPⅠ respectively.A large number of transgenic plants were obtained.The plasmid vcctor pEC23 carried a marker gene expressing Km resistance and the gene of tobacco mosaic virus 54KD protein, the latter controlled by CaMV35s promoter. The plasmid vector PRCPⅠ carried the marker gene expressing Km resistance and two other genes, cucumber mosaic virus(CMV) coat protein and CMV satellite RNA, both the latter were controlled by CaMV35s promoter. The leaf disks of cultivar G-28 and K326 were transferred separately on MS medium added 0.2 mg/l NAA, 2.0mg/l 6-BA,0.1mg/l KT and 60mg/l Km for G-28, 20mg/l for K326 after cocultivation with A, tumefaciens with the plasmid vectors. Shoots formed from transformed leaf disks in 2 or 3 weeks. Then the shoots were excised off and in MS medium added 0.2mg/l NAA 60mg/l Km for G-28 or 20mg/l for K326 to further select the Km resistant shoots and stimulate to form roots. Finally, the Km resistance of rooted plants was detected. The results indicated that the genes, at least the Km resistant marker gene were introduced into the plants.

关 键 词：烟草 烤烟 品种 抗病性 遗传 

分 类 号：S572.03[农业科学—烟草工业]