摘要
评估玻璃粉创伤棉胚茎尖分生组织的转化效率及初步建立其遗传转化体系.高速涡旋的玻璃粉创伤棉胚茎尖分生组织,利用农杆菌介导法将报告基因gusA导入棉花,GUS组织化学染色后观察棉胚转化情况,探讨不同转化条件对转化的影响.结果表明:0.2 g玻璃粉创伤棉胚,茎尖分生组织的较多细胞被转化,成活12株幼苗,较适宜.涡旋90 s转化条件下获得2株GUS阳性苗,涡旋时间越长茎尖分生组织创伤程度越严重,能提高转化效率,但成苗量降低.此外,转化效率受菌液浓度及菌种的影响,OD600为0.8的农杆菌LBA4404菌液侵染棉胚,较多细胞能表达gusA的活性且表达水平较高,而被农杆菌GV3101侵染后的棉胚染色效果较差.
Abstract
The study aimed to evaluate transformation efficiency of cotton shoot-tip meristem wounded by glass powder and establish the genetic transformation system.After mature embryos were wounded by high-speed whirling glass powder,gusA was transformed by Agrobacterium mediated method.The different factors that affected transformation frequency were discussed by the observation of GUS histochemical assay.The results showed that the suitable amount of glass powder was 0.2 g,with more cells of shoot-tip meristem transformed and 12 seedlings obtained.When the vortex time was up to 90 s,two GUS-positive plants were obtained.It showed that shoot-tip meristems were wounded more severely with the extended vortex time.Transformation efficiency was improved,but the number of seedlings was less.Additionally,transformation efficiency was affected by the concentration and the strain of Agrobacterium tumefaciens.More cells could highly expressed gusA gene activity after infested by Agrobacterium tumefaciens LBA4404 with OD600 =0.8,while Agrobacterium tumefaciens GV3101 was less effective.
NETL
NSTL
维普
万方数据