热带作物学报2011，Vol.32Issue(9)：1697-1703,7.DOI:10.3969/j.issn.1000-2561.2011.09.023
根癌农杆菌介导的木薯遗传转化条件的优化
Optimization of the Conditions for Agrobacterium-mediated Cassava Genetic Transformation
摘要
Abstract
As cassava genetic transformation is a complex process, it is pivotal to optimize relative conditions including explant culturing, infection and cocultivation by Agrobacterium, selection and plant regeneration of transformants. Among 11 farmer-preferred cassava varieties(KU50, NZ188, NZ199, SC5, SC6, SC8, SC124, TME7, TME12, Ebwanateraka and TMS60444)from Africa and China, different capacities for shoot organogenesis and somatic embryogenesis were noticed during the tissue culture processes. Most varieties could have a frequency of somatic embryogenesis above 60% with several more than 80%, such as TMS60444 and Ebwanateraka. Cultivars NZ199, SC124 and TME2 only gave very low frequencies, 25%, 43% and 36%, respectively. Shoot organogenesis could be induced from the cutting pieces of somatic cotyledons. After 2 weeks, somatic cotyledons produced shoot primordia and adventitious buds with a range of 15.8% to 75.8%. Among the tested cultivars, TME7 and TMS60444 performed the best. Similar to TMS60444, secondary somatic embryos of KU50, Ebwanateraka, SC5 and SC8 could form friable embryogenic callus after cyclic subculturing. Nevertheless, induction rates were very low in these cultivars in comparison with TMS60444. According to the results of cocultivation with different Agrobacterium tumefaciens strains and antibiotic dose responses, optimized conditions were established for the best transformation efficiency in cassava transformation. These conditions included infection with the modified Agrobacterium strain LBA4404/CS, the use of 200 u,m AS, cultivation temperature at 22 °C and cocultivation duration of 4 days. After cocultivation, antibiotic resistant somatic embryos should be regenerated under the selection containing 500 mg/L Carbenicillin and 15 mg/L of hygromycin. This study provides a protocol for condition optimization in cassava genetic transformation and will be useful for guiding the production of transgenic cassava using farmer-preferred cultivars.关键词
木薯/体细胞胚胎发生/芽器官发生/遗传转化/条件优化Key words
Cassava/ Somatic embryogenesis/ Shoot organogenesis/ Genetic transformation/ Condition optimization分类
农业科技引用本文复制引用
方佳敏,刘佳,唐克轩,张鹏..根癌农杆菌介导的木薯遗传转化条件的优化[J].热带作物学报,2011,32(9):1697-1703,7.基金项目
973计划项目(No.2010CB126605) （No.2010CB126605）
国家木薯产业技术体系项目(No.nycytx-17). （No.nycytx-17）
