两用核不育系P88S愈伤组织的诱导及其遗传转化研究

Callus Induction and Genetic Transformation for Dual Purpose Genic Male Sterile Line P88S

以超级杂交水稻两用核不育系P88S为材料,研究了其愈伤组织的诱导及植物抗病基因snc1在愈伤组织中的遗传转化。结果表明:P88S成熟胚愈伤组织的最适诱导和继代培养基是NMB培养基,其中2,4-D和6-BA的最适浓度分别为3.0 mg/L和0.2mg/L,而添加0.5 g/L的酸水解酪蛋白和脯氨酸有利于愈伤组织的诱导。P88S成熟胚愈伤组织的最适分化培养基是DL分化培养基,麦芽糖为抗性愈伤组织分化的最适碳源。并通过农杆菌介导遗传转化法共获得5株转化稻株,经PCR检测其中1株含目的基因snc1。

Taking super hybrid rice dual purpose genic male sterile line(DGMS) P88S as experimental material,its callus induction and genetic transformation of plant resistant gene sncl in its callus were researched.The results showed that the NMB is the optimum medium both for induction and subculture of P88S mature embryo callus,and the optimal concentration of 2,4-D and 6-BA are 3.0 mg/L and 0.2 mg/L,respectively;adding with 0.5 g/L of casein acid hydrolysate and 0.5 g/L of proline is beneficial to the callus induction;DL is the optimal differentiation medium for P88s mature embryo callus and maltose is the optimum carbon source for differentiation of resistant callus.Using the Agrobacteriummediate transformation method,five transformants are obtained and one of them is confirmed that containing the target gene snc1 by PCR test.