摘要
采用携带γ-TMT和bar基因表达载体pGBVE的根癌农杆菌(Agrobacteriumtumefaciens)菌 株GV3101对花生栽培品种鲁花11和丰花2号的成熟胚小叶片及胚生长点进行遗传转化。受体基因型及外 植体类型对转化效率影响很大,以鲁花11胚生长点的转化率最高。试验获得14株PPT抗性苗,经PCR检测 证明,γ-TMT基因已整合在花生基因组中。
Leaflets and apexes of embryo from two cultivars of peanut (Arachis hypogaea L.), Luhua11 and Fenghua No.2, were transformed with the strain of Agrobacterium tumefaciens GV3101 harboring expression vector pGBVE. The vector contained γ-TMT(γ-tocopherol methyltransferase) gene and bar gene. The precultured explants were infected with Agrobacterium tumefaciens GV3101(OD 600=0.65) for 15 min. After being co-incubated 4d, and continually cultured for regeneration and growth of buds, then regenerated shoots were transferred to selective medium. Most regenerated shoots turned to perish and then died;the transgenic ones grew normally, after selectively cultured two turns. The efficiencies of transformation and transgenic plant regeneration vary greatly with the influence of receptor genotypes and explant types. The highest transformation efficiency is obtained when apexes of embryo from cultivar Luhua11 are infected as explants.14 PPT-resistance plants are obtained. The integration of γ-TMT gene into the genome of transgenic plants is confirmed by PCR analysis.
出处
《中国粮油学报》
EI
CAS
CSCD
北大核心
2005年第1期61-64,68,共5页
Journal of the Chinese Cereals and Oils Association
基金
"十五"国家重点科技攻关项目(2001BA511B)
