含GmNHX1和LcVP1双价基因植物表达载体构建与转基因棉花的获得  

Construction of Plant Expression Vector with GmNHX1 and LcVP1 Dual-gene and Obtaining Transgenic Cotton

作　　者：冯雪[1] 贾永红[1] 郭红珍[1] 张一名[1] 赵学良[1] 孙艳香[1] 

机构地区：[1]廊坊师范学院遗传与育种研究所,河北廊坊065000

出　　处：《华北农学报》2014年第4期56-62,共7页Acta Agriculturae Boreali-Sinica

基　　金：河北省自然科学基金项目(C2010001804);河北省高等学校科学技术研究项目(20070318);廊坊师范学院博士基金项目(LSZB200903);廊坊师范学院自然科学重点项目(LSZZ201102)

摘　　要：应用DNA重组技术将编码大豆液泡膜Na+/H+反向转运蛋白GmNHX1和百脉根液泡膜H+-PPase LcVP1的开放阅读框片段同时构建在pCambia1301载体中,形成p1301-GmNHX1-LcVP1双价耐盐基因植物表达载体。以Coker 312棉花品种的胚性愈伤组织为外植体,采用根癌农杆菌介导法将其转入棉花中,并利用潮霉素筛选获得抗性植株。对抗性植株的T1幼苗进行分子和生理指标检测,表明外源基因已成功整合到棉花基因组中,并不同程度地提高了转基因株系耐盐能力。By use of DNA recombination method ,a vector,p1301-GmNHX1-LcVP1,harboring two open reading frames of soybean tonoplast Na＋/H＋antiporter GmNHX1 and Lotus corniculatus H＋-PPase LcVP1,was constructed through sequential restriction digests and ligations recombination .The two genes driven by CaMV 35 S promoters were suc-cessfully transferred into cotton cultivars Coker 312 via Agrobacterium with hygromycin selection system .The molec-ular and physiological analysises of T 1 generation indicated that genes were integrated into the cotton genome and enhanced transgenic cotton salt tolerance .

关 键 词：棉花 耐盐 大豆液泡膜Na+/H+反向转运蛋白GmNHX1 百脉根液泡膜H+-PPase LcVPl 遗传转化 

分 类 号：Q78[生物学—分子生物学] S562.03[农业科学—作物学]