苜蓿植株再生及PPRV-F基因转化条件的研究  

Optimization of Alfalfa Regeneration and Transformation Conditions with H gene of Peste des Petits Ruminants Virus

作　　者：杨国锋[1] 刘霞[2] 孙娟[2] 王志亮[3] 

机构地区：[1]青岛农业大学生命科学学院,青岛2661091 [2]青岛农业大学动物科技学院 [3]中国动物卫生与流行病学中心

出　　处：《青岛农业大学学报（自然科学版）》2010年第3期237-240,共4页Journal of Qingdao Agricultural University(Natural Science)

基　　金：山东省优秀中青年科学家科研奖励基金(BS2009SW010);青岛农业大学高层次人才启动基金(630717)

摘　　要：以苜蓿品种甘农1号下胚轴为外植体,研究不同培养基对苜蓿愈伤组织、体细胞胚形成的影响,通过体细胞胚发生途径建立再生体系,在此基础上,摸索农杆菌介导小反刍兽疫病毒F基因的转化条件。结果表明:所选培养基愈伤组织诱导率均为100%,UM+0.1mg/L NAA+0.5mg/L KT为诱导体细胞胚的最佳培养基。农杆菌介导的苜蓿遗传转化条件为:下胚轴预培养3d,农杆菌菌液侵染15min,然后在培养基上铺一层灭菌滤纸共培养3d后清洗,选择压为卡那霉素(Km)75mg/L,抑菌浓度为头孢霉素(Cef)300mg/L。本研究为制备防治小反刍兽疫转基因植物疫苗奠定了基础。Regeneration system of alfalfa cultivar Gan-nong No.1 was established through somatic embryogenesis.Callus and somatic embryo were induced by different medium and by hypocotyl as explants.Based on this system,Alfalfa transformation Condition by Agrobacterium-mediated with F gene of peste des petits ruminants virus（PPRV） was optimizated.The results showed that all of the probability of callus induced by different medium was 100%,and the best medium which can induce somatic embryo was UM＋0.1mg/L NAA＋0.5mg/L KT.The transformation system of alfalfa by Agrobacterium-mediated with F gene of PPRV was that pre-cultivated for 3 days,the hypocotyl was immersed in the bacterial suspension for 15 minutes and then cocultivated on MS medium covered with a piece of sterile filter paper for 3 days before washing,concentration of Km and Cef was 75mg/L and 300mg/L respectively.This study will lay the foundations for the preparation of transgenic plant vaccine which can prevent and cure PPR.

关 键 词：苜蓿 再生体系 小反刍兽疫病毒 遗传转化 

分 类 号：Q813.12[生物学—生物工程]