林业科学2013，Vol.49Issue(7)：62-68,7.DOI:10.11707/j.1001-7488.20130709
干旱胁迫下柠条锦鸡儿叶片SSH文库构建及CkWRKY1基因克隆
Construction of a Suppression Subtractive Hybridization Library of Caragana korshinskii Under Drought Stress and Cloning of CkWRKY1 Gene
摘要
Abstract
Caragana korshinskii is an important cultivated shrub with high ecological and forage value,and widely distributes in Northwest China.C.korshinskii has a strong tolerance to drought,cold,heat,saline and poor soil.In order to study its resistance mechanisms to drought and explore for drought response related genes,a suppression subtractive hybridization (SSH) library of C.korshinskii under drought stress was constructed.Totally 1 286 ESTs from the SSH library were obtained and the average length was 475 bp.As many as 645 Unigenes were identified through the EST sequence assembly.Many drought-related genes were annotated by Blasting,such as the genes encoding dehydration related LEA and DHN proteins as well as transcription factors including MYB,bZIP,WRKY,NAC,and bHLH families.A candidate gene encoding a member of WRKY transcription factor family was cloned by rapid amplification of cDNA ends technique,and was named as CkWRKY1 (GenBank accession No.JX987095).The protein deduced from the cDNA was composed from 330 amino acids with a conserved region "WRKYGQK".Real-time-quantitative PCR analysis showed that drought stress enhanced CkWRKY1 expression at the transcriptional level,indicating that CkWRKY1 might be involved in the drought resistance mechanisms of C.korshinskii at molecular level.关键词
柠条锦鸡儿/干旱胁迫/抑制性差减杂交(SSH)文库/WRKY/基因克隆Key words
Caragana korshinskii/ drought stress / suppression subtractive hybridization (SSH) library / WRKY / gene cloning分类
农业科技引用本文复制引用
杨杞,张涛,王颖,李高,尹佳佳,韩晓敏,齐力旺,李国婧,王瑞刚..干旱胁迫下柠条锦鸡儿叶片SSH文库构建及CkWRKY1基因克隆[J].林业科学,2013,49(7):62-68,7.基金项目
教育部博士学科点基金项目(20111515110001) （20111515110001）
内蒙古农业大学科技创新团队培育计划(NDPYTD2010-3). （NDPYTD2010-3）
