生菜遗传转化体系的建立及转基因研究

以散叶生菜大速生(Lactuca sativavar.capatataL.)为试材,以MS为基本培养基,采用不同激素配比,确定生菜高效诱芽培养基为MS 1.5mg/L6-BA 0.2mg/LIAA;抗生素敏感性试验表明,筛选培养基中适宜的潮霉素选择压为20mg/L,抑菌剂羧苄青霉素的适宜质量浓度为300mg/L;通过根癌农杆菌介导的叶盘法将携带O型和A型口蹄疫抗原决定簇融合基因O21-O14-A21-HBcAg转入大速生散叶生菜,对部分抗性植株进行PCR和PCR-Southern杂交检测,证实目的基因已经成功整合到生菜基因组中。RT-PCR检测初步表明,O21-O14-A21-HBcAg基因可以在生菜中正常转录表达。

Establishment of Genetic Transformation System and Transgenic Studies in Lettuce (Lactuca sativa var.capatata)

In this experiment,the effects of different hormone concentration on shoot regeneration from cotyledon explants are investigated.The results show that MS medium supplied with 1.5 mg/L 6-BA and 0.2 mg/L IAA is the suitable shoot regeneration.The experiment on sensitivity of cotyledon to hygromycin B and carbenicillin shows that the suitable hygromycin concentration for selecting transgenic tissue is 20 mg/L and the carbenicillin concentration is 300 mg/L.Fused gene O21-O14-A21 containing both type O and A FMDV is transferred into lettuce mediated by Ti plasmid of Agrobacterium tumafaciens.The results of PCR and PCR-Southern blotting indicated that the O21-O14-A21 gene were successfully integrated into the genomes of some transformed lettuce plants.Expression of O21-O14-A21 gene in transformed lettuce plants was proved by RT-PCR analyses.