上西早生柿ACC氧化酶的转基因研究  

Study on transformation of Uenishiwase Diospyros kaki Thunb with ACC oxidase gene

作　　者：刘艺[1] 马俊莲[1] 张子德[1] 唐霞[1] 宋春丽[2] 

机构地区：[1]河北农业大学食品科技学院,河北保定071001 [2]河北农业大学中兽医学院,河北定州073000

出　　处：《河北农业大学学报》2009年第2期66-70,共5页Journal of Hebei Agricultural University

基　　金：河北省国际科技合作攻关项目(04395501D-3)

摘　　要：为了得到耐贮藏的转基因柿,以上西早生柿叶片为外植体,通过农杆菌介导法,建立了ACC氧化酶的RNAi遗传转化体系。研究了预培养时间、农杆菌菌液浓度、侵染时间、乙酰丁香酮(AS)浓度和共培养时间等因素对基因转化的影响。结果表明:适宜的壮观霉素浓度为30 mg/L,预培养时间为3 d,用OD600值为0.3的菌液侵染10 min、AS浓度100 mg/L、共培养4 d有利于提高转化频率。经PCR分子检测与GUS组织化学染色,初步确定ACC氧化酶基因已转入上西早生柿组培苗中。In order to get transgenic Japanese Persimmon,using young leaves of Uenishiwase as ex- plant, ACC oxidase gene was transferred into Uenishiwase, mediated by Agrobacterium tumefa-ciens. The pre-cultured time, the concentration of A. tumefaciens, the infection time, the con- centration of AS and the co- cultured time were studied. Together studied was the genetic transfor- mation systems. The results showed that the available concentration of Spectinomycin was 10 mg/L, the best pre- cultured time was 3 d, the fittest OD600 of engineering bacterial suspension was 0.3, the optimum infection time was 10 min, the co- cultured time was 4 d and AS 100 mg/L could in- crease the frequency of transformation. It was originally proved by the PCR examination and GUS assays that the ACO gene had been transferred into Uenishiwase.

关 键 词：上西早生柿 根癌农杆菌 ACC氧化酶基因(ACO) 遗传转化 

分 类 号：S665.2[农业科学—果树学]