花生上胚轴为外植体的植株再生及转化研究  

Plantlet Regeneration and Agrobacterium-mediated Transformation of Peanut with Epicotyl as Explants

作　　者：万小荣[1] 杨丽霞[2] 李玲[2] 

机构地区：[1]仲恺农业技术学院生命科学学院,广州510225 [2]华南师范大学生命科学学院,广州510631

出　　处：《华中农业大学学报》2007年第1期15-19,共5页Journal of Huazhong Agricultural University

基　　金：广东省自然科学基金博士科研启动项目(06301202);仲恺农业技术学院博士启动项目(G2360225);广东省自然科学基金项目(06025049)资助

摘　　要：以8 d龄花生无菌苗的上胚轴为外植体,进行植株再生和农杆菌介导遗传转化研究。结果表明,上胚轴能高效诱导花生植株再生,外植体在MS+3 mg/L TDZ+6 mg/L6-BA+1.5 mg/L NAA培养基上能高效诱导不定芽分化,10 d时诱芽率达100%;不定芽接种到MS盐+B5维生素+3 mg/L TDZ+0.01%酵母粉培养基中15 d诱导出大量丛生芽;MS+0.5 mg/L NAA培养基较适合诱导丛生芽生根,25 d时生根率达82%。筛选确定35 mg/L潮霉素浓度为转化筛选压,以含AtRD29Ap∷GUS融合基因的根癌农杆菌侵染8 d龄上胚轴10 min,共培养3 d,经上述再生体系,以潮霉素筛选获得1株拟转化再生植株。The epicotyls of 8-d-old sterile peanut seedlings were used as explants to study plantlet regeneration and Agrobacterium-mediated genetic transformation. The results showed that the adventitious bud differentiation could be efficiently induced from the explants on MS solid medium supplemented with 3 mg/L TDZ,6 mg/L 6-BA and 1. 5 mg/L NAA. The budding frequency reached 100% after a 10-d-old culture of the explants on bud induction medium. Multiple buds were induced from the adventitious buds after a 15-d-old inoculation in the medium with MS salt, B5 vitamin, 3 mg/L TDZ and 0. 01% yeast powder. The optimum medium for root formation on the multiple buds was MS medium containing 0. 5 mg/L NAA. The rooting percentage added up to 82% after a 14-d-old culture of elongated shoots on root induction medium. The concentration of 35 mg/L hygromycin was used to screen the putative transformants in the following transformation of peanut mediated by Agrobacterium tumefaciens containing a binary vector harboring the drought-inducible promoter AtRD29Ap fusioned with GUS gene. Infected by the Agrobacterium for 10 min,the epicotyl explants were co-cultivated with the Agrobacterium for 3 d, and then were gultured on the above described regeneration system. One putative transformed regenerated plantlet was obtained after continuous selection for hygromycin resistance.

关 键 词：花生 上胚轴 植株再生 根癌农杆菌 遗传转化 

分 类 号：S565.203.53[农业科学—作物学]