玻璃粉创伤棉花茎尖分生组织遗传转化的研究  

Research on Genetic Transformation of Cotton Shoot-Tip Meristem Wounded by Glass Powder

作　　者：尹秀[1] 游义霞[1] 张维[1] 黄先忠[1] 崔百明[1] 

机构地区：[1]石河子大学生命科学学院/农业生物技术重点实验室,石河子832003

出　　处：《石河子大学学报（自然科学版）》2013年第6期736-741,共6页Journal of Shihezi University(Natural Science)

基　　金：转基因生物新品种培育科技重大专项(2011ZX08005-002)

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:0.2g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90s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。The study aimed to evaluate transformation efficiency of cotton shoot-tip meristem wounded by glass powder and es- tablish 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. Transfor- mation efficiency was improved, but the number of seedlings was less. Additionally, transformation efficiency was affected hy the concentration and the strain of Agrobacterium tumefaeiens. More cells could highly expressed gusA gene activity after infested by Agrobacterium tumefaciens LBA4404 with ODs00 = 0.8, while Agrobacterium tumefaciens GV3101 was less effective.

关 键 词：棉花 农杆菌介导转化 茎尖 遗传转化 

分 类 号：S562.035[农业科学—作物学]