甘露糖阳性选择系统的建立及在番茄转化中的应用  

A Novel Selection System for Tomato Transformation by Mannose

作　　者：彭世清[1] 陈守才[1] 

机构地区：[1]中国热带农业科学院热带生物技术研究所热带作物生物技术国家重点实验室,海口571101

出　　处：《农业生物技术学报》2005年第2期141-144,共4页Journal of Agricultural Biotechnology

基　　金：中国热带农业科学院科技基金资助项目

摘　　要：利用PCR克隆了大肠杆菌(Escherichiacoli)6-磷酸甘露糖异构酶基因(pmi)并进行了序列测定。将该基因替换植物表达载体pCAMBIA2301中的卡那霉素抗性基因,构建了以pmi为选择标记基因的植物表达载体pPMI,并导入根癌农杆菌(Agrobacteriumtumefaciens)EHA105中。研究了甘露糖对番茄(Lycopersicumesculentum)生长和生根的影响及叶盘对甘露糖的敏感性。通过根癌农杆菌介导对番茄进行遗传转化,获得转化番茄再生植株,并对转化植株进行了PCR扩增检测。研究表明以甘露糖阳性选择系统在番茄遗传转化中应用是可行的。The 6-phosphomannose isomerase gene (pmi ) of Escherichia coli was amplified by PCR. Based on pCAMBIA2301, the plant expression vector with pmi was constructed by substituting pmi for kanamycin resistance gene. Effects of mannose on tomato germination and growth were examined. A novel selectable system using in Agrobacterium-mediated transformation of tomato by mannose was developed. The selection condition was established by mixing mannose (0~5 g/L) and sucrose (30 g/L). The last selection pressure on shoot formation was created by a mixture of sucrose and mannose at 30 g/L and 3 g/L. Stable transformation was confirmed by PCR.

关 键 词：选择系统 甘露糖 番茄 应用 阳性 卡那霉素抗性基因 植物表达载体 根癌农杆菌介导 选择标记基因 遗传转化 PCR克隆 PCR扩增 大肠杆菌 序列测定 基因替换 再生植株 转化植株 酶基因 pmi 敏感性 导入 生根 

分 类 号：S641.2[农业科学—蔬菜学] F273.1[农业科学—园艺学]