摘要
以水稻两用不育系培矮64S成熟胚为外植体,以不同组合培养基为诱导和继代培养基,建立了适合水稻培矮64S转化的高效再生体系,并通过农杆菌EHA105介导,将大肠杆菌C1基因转入培矮64S,获得再生植株。结果表明,J0N1D3为合适的愈伤组织诱导培养基,诱导率达56.44%;通过潮霉素筛选后,获得的抗性愈伤组织分化率为73.08%,经分化,获得133株再生植株;随机挑选67株再生植株经PCR检测,其中46株检测到目的条带,阳性检出率占68.66%;对PCR阳性植株进行荧光定量实时PCR分析,结果表明C1基因能在转基因植株中表达。通过对T1代PCR分析,得到目的条带,表明C1基因能在转基因后代稳定遗传。
A high efficient regeneration system of rice plants for genetic transformation was developed through optimizing culture media,when mature embryos of Pei＇ai 64S were used as explants. CI gene cloned from E. coil was successfully delivered into Pei＇ai 64S through the Agrobacterium-mediated method, and stable transgenic plants were obtained. The rate of callus induction reached 56.44% using J0N1D3 as the induction medium. A total of 133 hygromycin resistant plants were obtained with a regeneration rate of 73.08%. A group of 67 of them were chosen randomly for PCR analysis, and 46 were positive for the gene （68.66%）. Quantitative RT-PCR analysis showed that the expression of exogenous C1 gene in transgenic rice plants. PCR analysis of T1 plants showed that the integrated C1 gene could be steadily inherited.
出处
《农业现代化研究》
CSCD
北大核心
2009年第3期364-368,共5页
Research of Agricultural Modernization
基金
中国科学院“百人计划”项目(编号:02200420062903)
中国科学院亚热带农业生态研究所青年人才领域前沿项目(编号:ISACX-LYQY-QN-0706)
关键词
水稻
培矮64S
成熟胚
根癌农杆菌
转化体系
组织培养
rice
Pei＇ ai 64S
mature embryo
Agrobactenum tumefaciens
transformation system
tissue culture
