陆地棉(G.hirsutum L.)N-乙酰氨基葡萄糖转移酶基因(GhGnT)的克隆及耐盐性功能分析  

Cloning and Salt-tolerance Analysis of N-acetylglucosaminyltransferase Gene (GhGnT) from Gossypium hirsutum L.

作　　者：周凯[1] 宋丽艳[1] 王俊娟[1] 王德龙[1] 樊保香[1] 王帅[1] 叶武威[1] 

机构地区：[1]中国农业科学院棉花研究所,棉花生物学国家重点实验室,农业部棉花遗传改良重点实验室,安阳455000

出　　处：《分子植物育种》2011年第3期309-317,共9页Molecular Plant Breeding

基　　金：973项目(2010CB126006);转基因生物新品种培育科技重大专项(2008ZX08005)共同资助

摘　　要：在耐盐相关基因芯片基础上,我们筛选到盐诱导显著上调表达(Ratio>2)的N-乙酰氨基葡糖转移酶基因。选取耐盐材料中9806为材料,根据耐盐性抑制消减文库EST序列设计引物,利用RACE及RT-PCR技术获得该基因cDNA全长1970bp,命名为GhGnT。通过Blast比对,发现该基因与蓖麻乙酰氨基葡萄糖转移酶基因相似性最高,达到82%,认为GhGnT与该基因属于同类基因。利用TargetP1.1和toppred分析,GhGnT蛋白跨线粒体膜。由Real-time分析发现,该基因受盐胁迫诱导上调表达,在耐盐材料中9806的表达水平明显高于盐敏感材料中S9612,与芯片结果一致。有研究发现在小麦中发现糖基转移酶TaUGT1和TaUGT2受盐诱导上调表达,而且转化糖基转移酶亚基STT3a基因的杜氏盐藻耐盐性得到提高。因此,我们认为该基因与耐胁迫密切相关,并推测GhGnT的高表达对提高陆地棉耐盐性有一定作用。GhGnT基因的克隆及分析为以后的功能验证及耐盐种质的创新奠定基础。Based on salt-tolerant gene chips, a N-acetylglucosaminyltransferase gene encoding resistance of salinity has been screened by ratio index （Ratio〉2）. Zhong9806, salt resistance spp. of Gossypium hirsutum L. was used as experimental materials, and primers were designed on the basis of EST from salt resistance related SSH library. The full cDNA, 1 970 bp long, was gained by using RACE and RT-PCR technology and the gene was named as GhGnT, which shows, by Blast comparison the highest simility, 82%, as the acetylglucosaminyltransferase gene of Ricinus communis. Its protein acrossed the mitochondrial membrane through analyzed by TargetP 1.1, and its expression raised under salt stress induction and the expression level of GhGnT of Zhong9806 （salt-resistance material） was obviously higher than ZhongS9612 （salt-sensitive material） by Real-time PCR, which was in accord with the arrays results. It has already found that glycosyltransferase TaUGT1 and TaUGT2 transforming glycosyltransferase and subunitting STT3a gene to improve salt resistance were overexpressively induced by salt on wheat. So we believe the gene was bound up with salt stress and assumed that the overexpression of GhGnT do good to improve salt-tolerance of Gossypium hirsutum. Cloning and analysis of GhGnT gene lay the foundation for the functional verification and salt resistance germplasm innovation.

关 键 词：基因芯片 乙酰氨基葡萄糖转移酶 Real-time 耐盐 陆地棉 

分 类 号：S562[农业科学—作物学]