百脉根高频再生体系的建立及兔出血症病毒衣壳蛋白VP60基因的转化  

Establishment of High Efficient System on Lotus Regeneration and Transformation of Capsid Protein (VP60) Gene of Rabbit Hemorrhagic Disease Virus in Lotus corniculatus

作　　者：唐广立[1] 李传山[1] 陈明利[1] 张忠品[1] 郭蔼光[1] 

机构地区：[1]西北农林科技大学生命科学学院

出　　处：《分子植物育种》2007年第4期593-600,共8页Molecular Plant Breeding

基　　金：西北农林科技大学博士创新基金资助。

摘　　要：以MS为基本培养基,通过调整激素种类与浓度等培养条件,按正交试验设计的原则,建立了百脉根高频再生体系。结果表明MS+2,4-D2.0mg/L+KT2.0mg/L培养基可高效诱导愈伤组织的形成,MS+6-BA1.0mg/L+NAA0.1mg/L培养基可高效诱导芽的分化,MS+NAA0.1mg/L培养基可快速诱导根的生成,形成再生植株。通过构建植物表达载体VP60-pBI121,研究了根癌农杆菌介导的兔出血症病毒(RHDV)衣壳蛋白VP60基因对百脉根遗传转化的影响因素,建立了百脉根快速高效遗传转化体系,结果表明,以农杆菌LBA4404为介导菌株、以下胚轴为外植体,预培养3d,在OD600为0.6的菌夜中浸染20min,共培养3d,以及300mg/L羧苄青霉素脱菌浓度和50mg/L卡那霉素筛选浓度为最佳转化条件。为利用百脉根生产动物口服型疫苗建立了技术基础。In this paper, MS was selected as the base culture medium. According to the orthogonal design principle, the high-efficient regeneration system of Lotus corniculatus Linn were found through adusting the varieties and concentration of hormones. The result showed that the medium of MS＋2,4-D 2.0 mg/L ＋KT 2.0 mg/L could effi- ciently induce callus and the medium of MS＋6-BA 1.0 mg/L ＋NAA 0.1 mg/L could efficiently induce bud of leaf, the medium of MS＋NAA 0.1 mg/L induced the regeneration of roots quickly, and the regenerated plant formed. Vector VP60-pBI121 was constructed and transformed to Lotus corniculatus Linn by Agrobacterium tumefaciens. The regenerated resistant plants could be obtained by using hypocotyls transformation recipient, infecting the 3-day pre-culturing hypocotyls with Agrobacterium tumefaciens LBA4404 suspension （OD600≈0.6） for 20 min, after a 3-day co-cultivation period transferring to the callus inducement media containing 300 mg/L Carbenicillin and 50 mg/L Kanamycin for 20 d. This system is the elementary technology for production of mammal edible vaccine.

关 键 词：百脉根(Lotus comiculatus Linn) 高频再生体系 兔出血症病毒(RHDV) 转化 

分 类 号：S541.6[农业科学—作物学]