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利用根癌农杆菌(Agrobacteriumtumefaciens(SmithetTownsend)Conn)介导法将含有豌豆外源凝集素(pealectin,PLec)基因和大豆Kunitz型胰蛋白酶抑制剂(soybeanKunitztrypsininhibitor,SKTI)基因的双价抗虫基因植物表达载体pBinLK用于陆地棉(GosypiumhirsutumL.)栽培品种“新陆早1号”、“新陆中2号”、“冀合321”和“辽9”的转化。棉花无菌苗下胚轴经过与根癌农杆菌共培养、卡那霉素抗性愈伤组织的筛选、体细胞胚状体的诱导和植株再生等阶段成功地获得了双价抗虫基因陆地棉转化植株。NPTⅡ的ELISA检测、PCR鉴定和PCRSouthern检测证实,两个外源抗虫基因同时存在于再生植株基因组内。抗虫测试结果表明,转基因棉株对棉铃虫(HeliothisarmigeraHubner)幼虫具有较强的抗性
The bivalent insect-resistant genes containing pealectin (PLec) gene and soybean Kunitztrypsin inhibitor (SKTI) gene were amplified by Agrobacterium tumefaciens (Smithetownown) The plant expression vector pBinLK was used for the transformation of Gossypium hirsutum L. cultivars “Xinluzao 1”, “Xinluzhong 2”, “Jihe 321” and “Liao 9”. Cotton aseptic seedlings hypocotyls after co-cultivation with Agrobacterium tumefaciens, kanamycin-resistant callus screening, somatic embryoid body induction and plant regeneration stage successfully obtained bivalent insect-resistant genes land Cotton transformed plants. NPT Ⅱ ELISA detection, PCR identification and PCR Southern detection confirmed that the two exogenous insect-resistant genes exist in the regenerated plant genome. Insect-resistant test results showed that the transgenic cotton plants had strong resistance to Heliothis armigera Hubner larvae