农杆菌介导rd29A启动子驱动otsB基因转化紫茉莉的研究  

Transformation of Mirabilis jalapa L. with ots B Gene in E.coli Driven by rd29A Promoter from Arabidopsis thaliana

作　　者：陆玉建[1,2,3] 张韩杰[3] 韩文瑜[2] 沈志强[1,4] 

机构地区：[1]山东省滨州畜牧兽医研究院/博士后科研工作站,滨州256600 [2]吉林大学博士后科研流动站,长春130062 [3]滨州学院生命科学系/山东省黄河三角洲野生植物资源开发利用工程技术研究中心,滨州256603 [4]山东省滨州畜牧兽医研究院,滨州256600

出　　处：《植物研究》2016年第3期401-408,共8页Bulletin of Botanical Research

基　　金：山东省自然科学基金(ZR2012CL14);滨州学院博士基金(2010Y08)

摘　　要：紫茉莉为紫茉莉科紫茉莉属多年生草本植物,具有较高观赏和药用价值,对重金属和石油污染有较强修复能力。然而目前为止,紫茉莉再生和遗传转化体系尚未建立。本研究以紫茉莉为材料,初步建立起较为稳定的再生体系。通过克隆拟南芥rd29A启动子和大肠杆菌ots B基因,构建了p2300-rd29A pro-ots B植物表达载体。利用根癌农杆菌介导法对紫茉莉进行遗传转化。结果表明,当农杆菌的OD600=0.5,侵染成熟胚或带芽茎段60 min,共培养2 d,紫茉莉的转化效率较高。通过对筛选出的转基因植株进行PCR检测,显示大肠杆菌ots B基因已成功整合到紫茉莉的基因组中,并可有效的进行转录。Mirabilis jalapa L. is a perennial herb of Nyctaginaceae. Not only is it a high-value ornamental flowering and greening plants,but also can be used as medicine. M. jalapa L. has good bioremediation function. M. jalapa L. was used as experimental material,and the perfect regeneration system was initially established. The rd29 A promoter of Arabidopsis thaliana and ots B gene in E. coli were cloned by PCR respectively. Afterwards,the rd29 A promoter and ots B gene were ligated into plasmid p2300-GFP,which would lead to the construction of p2300-rd29 A pro-ots B expression vector. Then,the expression vector was introduced into the cells of M. jalapa L. by Agrobacterium-mediated method. The genetic transformation results showed that when the concentration of Agrobacterium was OD600= 0. 5,the time of infection of mature embryo or nodal stem segments was 60 min,and the co-culture time was 2 d,the transformation efficiency of M. jalapa L. was the highest. By PCR detection,ots B gene was successfully integrated into the genome of M. jalapa L.,and the efficient transcription could be performed.

关 键 词：紫茉莉 RD29A启动子 otsB基因 农杆菌 遗传转化 

分 类 号：S685.16[农业科学—观赏园艺]