华南农业大学学报2013，Vol.34Issue(3)：378-382,5.
农杆菌子房注射法对金钗石斛的活体转化研究
In Planta Transformation of Dendrobium nobile by Ovary-Injection of Agrobacterium
摘要
Abstract
Dendrobium nobile was transformed by ovary-injection of Agrobacterium.After artificial pollination in 10,20,30,40,45,50 and 60 d,EHA105 (pCAMBIA1301) harboring both β-glucuronidase gene (GUS) and hpt was injected into the ovary.The seeds of split fruit of D.nobile were cultured on the N16 culture medium by aseptic processing and detected by GUS expression.Forty-five days after cultured,the protocorms were diverted to N16 medium with hygromycin to gain hygromycin-resistance protocorms,then four randomly selected hygromycin-resistance protocorms were detected by PCR analysis.The study showed that ovary-injection of agrobacterium had harmful effects on the fruit development and seed formation.Ovary-injection of agrobacterium after artificial pollination 45 d had the highest efficiency,18％ seeds from the injected position showing GUS expression.Eleven hygromycin-resistant plants were gained in this study,and four randomly selected hygromycin-resistant plants were detected by PCR analysis and all of them were positive.The results showed that ovary-injection of agrobacterium transformation had advantages in genetic transformation of orchids.关键词
金钗石斛/农杆菌子房注射法/GUS组织化学染色/活体转化/PCR检测Key words
Dendrobium nobile / ovary-injection of Agrobacterium/ GUS assay / in planta transformation /detection of polymerase chainreaction分类
农业科技引用本文复制引用
刘其府,董会,曾宋君,陈之林,吴坤林,张建霞,段俊..农杆菌子房注射法对金钗石斛的活体转化研究[J].华南农业大学学报,2013,34(3):378-382,5.基金项目
国家自然科学基金(31160400) （31160400）
