影响农杆菌介导的小麦遗传转化条件的研究.PDFVIP

: 1 1 1 1 2 , , , , ( 1. , 453003; 2. , 571737) : 利用小麦成熟胚愈伤组织的高效再生体系, 对影响农杆菌介导的小麦遗传转化条 : 预培养时间共培养时间抗 生素( Kan) 筛选压头抱霉素浓度菌液浓度和侵染时间等进行了研究最后确立的转化条 为: 预培养时间为 1~ 2 , 共培养时间为 3 , K an筛选压为 100 mg /L, 头孢霉素所用浓度 500m g/L, 当菌液浓度 OD600 = 0. 6浸染 45m in并对得到 的再生植株进行了 PCR 检测, 初步证实了目的基因转到了小麦基因组中 : ; ; : S 512. 1 : A : 2010) Stu y on Genetic T ransformation Con itions ofW heatM e iate by Agrobacterium Tum efaciens 1 1 1 1 2 HE Jie, WANGW ei, HU Haiyan , ZHAO Junjie, XIAO Xianzhou ( 1. H enan Institute of Science an T echnology, X inxiangH e nan 453003, China; 2. Research Institute, Ch ineseA ca emy of Tropical Science, Danzhou H ainan 571737, Ch ina) Abstract: Callus in uction regeneration system ofw heat, genetic transformation ofw heat using an agrobacteri umme iate approach w ere applie in the research to stu y the genetic transformation con itions of w heat me iate by agrobacterium tum efaciens. The factors affecting the transformation, such as preculture , coculti vation, the selective pressure of antibiotic, Cef on the regeneration, agrobacterium concentration an infection perio w ere exam ine . The optmi ize proce ure w as as follow ing: after preculture for 1 to 2 ays, the callus in uction w ere ippe into an a agrobacterium wh ich concentrationw as OD600 0. 6 for 15 m in, then cocultiva tion for 3 ays, the exp lantsw ere transferre to selectiveme ia w ith 100 mg/L Km an 500 mg/L Ce.f These con itionsw ere benefite to mi proving the transform ation efficiency. PCR analysis show e that objective gene w ere transferre to wheat genome. Key wor s: wheat; agrobacterium tumefaciens; genetic transforma