湖南农业大学学报（自然科学版）2013，Vol.39Issue(5)：471-477,7.DOI:10.3724/SP.J.1238.2013.00471
农杆菌介导的遗传转化籼稻Kasalath的条件优化
Study on the method for effecient and rapid genetic transformation of Kasalath mediated by Agrobacterium
摘要
Abstract
At present,Kasalath is a model Indica material for rice transgenic research.By using the technique of plant tissue culture and Agrobacterium-mediated genetic transformation,we optimized the condition for the callus induction and the performance of genetic transformation of Kasalath.The results showed that immature or newly harvested mature seeds as explants were more effective to generate callus than the seeds stored for 1 year.Among the induction media with 0.5,1,1.5,2,2.5,3,10,20 or 30 mg/L 2,4-D solutions,the medium with 2.5 mg/L 2,4-D had the best performance in callus induction; in callus infection by co-culturing with Agrobacterium Ag10 or EHA105 solution with OD600nm being 0.05,0.1,0.15,0.2,0.3 or 0.4,the efficiency of transformation reached the optimum when OD600 nm of the working solution of both Ag 10 and EHA105 was 0.15,but EHA105 was superior to Ag10,its hygromycin-resistant callus reached up to 82.0％ and 3 d co-culture with EHA105 was perfect.When the resistant calli suffered rapid dehydration for 6-8 h or slow dehydration for 2-3 d,the regeneration frequency was evidently improved and the time needed for regeneration was shortened; while pre-culture before callus infection and pre-differentiation processing before hygromycin-resistant callus getting onto differentiation culturing had little effect on genetic transformation efficiency,so these procedures could be omitted.By the optimized procedures,the overall time from callus induction to regeneration of transformant plantlets was shortened to 57-60 d,and 65％ of transformation efficiency was attained.关键词
籼稻/2,4-D/农杆菌介导/遗传转化/影响因素Key words
Indica rice/ 2,4-D(2,4-dichlorophenoxyacetic acid)/ Agrobacterium-mediated/ genetic transformation/influence factors分类
农业科技引用本文复制引用
周艳,黄亚萍,周遥,吴亚先,周兰英,姜孝成..农杆菌介导的遗传转化籼稻Kasalath的条件优化[J].湖南农业大学学报（自然科学版）,2013,39(5):471-477,7.基金项目
国家自然科学基金项目(31071487 （）
31270348) （）
湖南省生态学重点学科建设项目(0713) （0713）
