GFP基因在苜蓿转化组织中的荧光表达量分析  

Analysis of the fluorescence expression of GFP gene in transformed alfalfa tissues

作　　者：韩斌[1] 马晖玲[1] 李云霞[1] 

机构地区：[1]甘肃农业大学草业学院草业生态系统教育部重点实验室中-美草地畜牧业可持续研究中心,甘肃兰州730070

出　　处：《草原与草坪》2010年第6期65-68,共4页Grassland and Turf

基　　金：甘肃省农业生物技术研究与应用开发项目-紫花苜蓿抗病转基因植株的选育(GNSW-2007-03);国家科技支撑计划"奶业专项"子课题1"优质高产抗逆苜蓿和饲料作物新品种选育"子专题(2006BAD04A04-01-08)资助

摘　　要：利用农杆菌介导法将溶菌酶(Lyz)与绿色荧光蛋白(GFP)基因转入"甘农3号"紫花苜蓿(Medicago sativa)中,获得含有双元基因的紫花苜蓿转基因植株。在荧光显微镜下对不同处理、不同培养时期的苜蓿转化愈伤组织及再生转化植株进行荧光检测和荧光表达量的对比分析。结果表明:经预培养的转化愈伤组织中的荧光表达量明显高于未经预培养的转化子;3 d为共培养的最佳时间;侵染时经过摇床上摇动的负压处理,荧光表达量高于静止状态;共培养3 d后的转化荧光表达量达最强,随培养时间延长,表达量少量减少,但能够稳定表达。The callus containing dual genes of Lyz-GFP genes were obtained as hypocotyls of alfalfa（Gan-nong No.3）were used as receptors respectively for transformation of Lyz-GFP genes by Agrobacterium tumefaciens mediated.Fluorescence detection and fluorescence expression of transformed alfalfa callus and regenerative plants of different treatment and different stages were comparative analyzed by fluorescence microscope.The results showed that the fluorescence expression of pre-cultured transformed callus was significantly higher than the transformed callus without pre-culture after the infection of Agrobacterium.Three days were the best time for co-cultivation;The quantity of fluorescent expression was higher than static state;Throught negative pressure treatment on shaking bed in infection period.The fluorescence expression of explants co-cultured after 3d reached the strongest;The expression decreased a little but it could express stably with incubation time extend.

关 键 词：苜蓿 绿色荧光蛋白GFP基因 荧光检测 

分 类 号：S668.403.5[农业科学—果树学] S54[农业科学—园艺学]