摘要
为研究棉花HSP70基因在转基因植物中抗旱效果,利用前期克隆的Gh HSP70基因,以p CAMBIA1304质粒为植物表达载体,构建了Gh HSP70基因的植物表达载体p CAMBIA1304-HSP70;采用冻融法转入根癌农杆菌EHA105菌株,通过叶盘法对模式植物烟草进行遗传转化研究。结果表明,在潮霉素(50 mg/L)选择压力下获得的烟草转化不定芽和完整植株,经过PCR方法以及GUS基因组织化学法检测鉴定转基因烟草,其中有5株为阳性植株。初步证实了Gh HSP70基因已导入烟草基因组中。为进一步研究该基因的功能提供实验基础,同时为后续转化棉花改善其抗旱能力奠定基础。
In order to study the response of cotton HSPTO gene to drought stress in transgenic plants, the pCAMBIA1304-HSPTO, plant expression vector, was constructed by using the cloned GhHSPTO gene and the pCAMBIA1304 vector. The pCAMBIA1304-HSPTO was transformed into EHA105 strain by freeze-thaw method. Then, the genetic transformation of tobacco was studied through leaf dish transformation method. Results showed that the tobacco adventitious buds and complete plants were obtained under Hygromycin （50 rag/L） selective pressure, the transgenic tobaccos were detected by GUS gene histochemical method and PCR method, and five of them were positive plants. We preliminarily confirmed that the GhHSPTO gene had transferred into tobacco genome. This study provided a fundamental experiment condition for further research on the GhHSPTO gene, and it might lay the foundation for genetic transformation in order to improve the drought resistance of cotton.
出处
《中国农学通报》
2015年第31期194-198,共5页
Chinese Agricultural Science Bulletin
基金
国家自然科学基金地区科学基金项目"小麦非生物胁迫相关MYB转录因子TaSIM的功能及分子机理研究"(31360264)
新疆维吾尔自治区自然科学基金资助项目"棉花干旱胁迫相关Hsp70基因的克隆及功能分析"(2014211A025)
"棉花抗逆相关MYB转录因子的功能分析"(2013211B19)
棉花生物学国家重点实验室开放课题"棉纤维品质相关MYB转录因子的功能分析"(CB2015A26)
新疆维吾尔自治区高技术研究发展计划"棉纤维发育相关基因表达分析及鉴定
挖掘与利用研究"(201411103)
