盐碱胁迫下羊草消减文库的构建及分析  

Construction and Analysis of Suppression Subtractive Hybridization Library of Leymus chinensis under Saline-alkaline Stress

作　　者：解莉楠[1] 聂玉哲[1] 张晓磊[1] 李玉花[1] 

机构地区：[1]东北林业大学生命科学学院,哈尔滨150040

出　　处：《分子植物育种》2007年第3期371-376,共6页Molecular Plant Breeding

基　　金：863计划(2001AA246102)资助。

摘　　要：以盐碱胁迫下的羊草地上部分cDNA为实验方（tester）,非盐碱条件下生长的羊草地上部分cDNA为消减方（driver）,利用抑制性消减杂交技术（suppression subtractive hybridization,SSH）构建了盐碱胁迫下羊草消减文库。从消减文库中筛选1920个阳性克隆,PCR鉴定插入片段大部分集中在300～800bp之间。将聚类后得到的552个非重复序列与GenBank中的序列进行比对,其中有548条与数据库中的序列有同源性,其中功能已知的339个,功能未知序列209条。获得了与盐碱胁迫相关的基因,如单脱氢抗坏血酸还原酶（MDAR）、γ-谷氨酰半胱氨酸合成酶（γ-ECS）、铁蛋白（Ferrtin）、水孔蛋白（PIP）、脱水素基因（dehydrin）、14-3-3蛋白等。本实验为抗逆基因的克隆及系统研究盐碱胁迫下羊草相关基因的表达奠定了基础。Using cDNA from Leymus chinensis seedlings grown on the natural habitat where is the saline-alkaline soil as tester and cDNA from seedlings in normal growth as driver, suppression subtractive hybridization （SSH） was employed to construct cDNA subtracted library. Of the 1 920 cDNA clones showed high quality sequences. The size of inserts was 300-1 000 base pair as it was determined by PCR. In the subtractive library, 552 clones were non-redundant, of which 339 had homology to the known-transcripts, and 209 to unclassified proteins. The remaining 4 ESTs had no homology in the databases searched. Sequence comparison of these clones using BLASTX led to the identification of several putative saline-alkaline stress related genes, such as MDAR, γ-ECS, Ferritin, PIP, dehydrin, 14-3-3. The putative ESTs will serve as useful resource to understand the molecular basis of saline-alkaline stress related genes and the further studying gene expression in Leymus chinensis.

关 键 词：盐碱胁迫 羊草 抑制性消减杂交 

分 类 号：S543.9[农业科学—作物学]