CPB-LAR-GFP表达载体的构建及其在紫花苜蓿愈伤组织中的表达

摘要

以无色花青素还原酶编码基因LAR为靶序列,构建携带绿色荧光蛋白GFP和抗除草剂bar基因的双标记选择植物表达载体,通过农杆菌介导法对紫花苜蓿进行遗传转化,经PCR检测和PPT筛选出抗性愈伤组织,转基因紫花苜蓿愈伤组织的缩合单宁含量比对照高30.8%.试验证明重组基因已转化至紫花苜蓿愈伤组织中并发挥生理功能,为研究缩合单宁合成与代谢的分子调控机制和培育具有抗除草剂和抗臌胀病的牧草新品种奠定基础.%Leucoanthocyanidin reductase is a key enzyme in plant condensed tannins synthesis.The target was Leucoanthocyanidin reductase encoded by LAR gene,the expression vector was constructed with the double-marker expressing green fluoresce protein GFP gene and herbicide resistance bar gene.Transgenic alfalfa callus were obtained by Agrobacterium tumefaciens mediated transformation,and the resistant callus were screened by PPT.The condensed tannins content in transgenic alfalfa callus was 30.8% higher than that of control.The recombinant gene had been transformed to alfalfa callus and played a physiological function.These research work will provide the fundamental information and the method for studying the regulation mechanism of condensed tannins synthesis and further alfalfa breeding through transgenic technology with character of resistance to herbicide and bloat disease.