二色补血草LbGRP基因的克隆及抗逆能力分析  

Cloning and stress tolerance analysis of an LbGRP gene in Limonium bicolor

作　　者：潘妍[1] 王玉成[1] 张大伟[1] 杨传平[1] 

机构地区：[1]东北林业大学林木遗传育种与生物技术教育部重点实验室,哈尔滨150040

出　　处：《遗传》2010年第3期278-286,共9页Hereditas（Beijing)

基　　金：黑龙江省国际合作项目(编号:WB07N02);黑龙江省青年基金专项(编号:QC07C56)资助

摘　　要：富含甘氨酸RNA结合蛋白(Glycine-rich RNA-binding proteins,GRP)是植物中重要的转录后调控蛋白,在植物的生长发育及对胁迫的抗性调控等过程中起重要作用。文章从二色补血草(Limonium bicolor(Bunge)O.)cDNA文库中克隆出了富含甘氨酸RNA结合蛋白基因(LbGRP)的全长cDNA序列(GenBank登录号:GQ398238)。为了研究LbGRP的抗逆功能,将其构建到酵母表达载体pYES2中,转化到酿酒酵母INVSc1菌株中,获得重组酵母INVSc1(pYES2-LbGRP),同时将转空pYES2质粒的酵母INVSc1(pYES2)作为对照。对重组酵母INVSc1(pYES2-LbGRP)和对照INVSc1(pYES2)进行NaCl、KCl、NaHCO3、Na2CO3、干旱和冷冻胁迫,比较它们在不同胁迫下的存活率。结果表明,INVSc1(pYES2-LbGRP)在各种胁迫下的存活率明显高于对照INVSc1(pYES2),证明LbGRP基因具有抗NaCl、KCl、NaHCO3、Na2CO3、干旱和冷冻等胁迫的能力,推测该基因参与了二色补血草的抗逆调控过程。Glycine-rich RNA-binding proteins （GRP） are involved in post-transcriptional regulation,and play important roles in plant growth,development and stress response.In the present study,the full-length cDNA of a GRP gene （LbGRP,GenBank accession number： GQ398238） was cloned from a cDNA library of Limonium bicolor.To investigate the stress tolerance of LbGRP,the recombinant plasmid pYES2-LbGRP was constructed by inserting the LbGRP gene into the yeast expression vector pYES2.The recombinant plasmid pYES2-LbGRP was transformed into yeast Saccharomyces cerevisiae INVSc1,and the INVSc1 strain transformed with empty pYES2 was used as the control.The recombinant yeast INVSc1 harboring LbGRP （pYES2-LbGRP） and the control INVSc1 harboring empty pYES2 were treated with NaCl,KCl,NaHCO3,Na2CO3,drought and frezzing,respectively,and their survial rates were compared under these stress conditions.The results showed that the survival rates of the recombinant yeast INVScl （pYES2-LbGRP） were higher than that of the control strain under these stress conditions,indicating that the LbGRP is tolerant to NaCl,KCl,NaHCO3,Na2CO3,drought and frezzing stresses.These results suggested that LbGRP plays a role in stress tolerance of L.bicolor.

关 键 词：二色补血草 富含甘氨酸RNA结合蛋白 抗逆 酵母表达 

分 类 号：S688.4[农业科学—观赏园艺] Q943[农业科学—园艺学]