干旱胁迫下新源1号新疆野苹果抑制性差减文库的构建与初步分析  

Construction and Preliminary Analysis of cDNA Library of Malus sieversii under Drought Stress by Suppression Subtractive Hybridization

作　　者：徐彦[1] 钟翡[1] 廖康[2] 李芳[1] 王惠庭[1] 李天红[1] 

机构地区：[1]中国农业大学农农学与生物技术学院果树系,北京市果树逆境生理与分子生物学重点开放实验室,北京100193 [2]新疆农业大学林学与园艺学院,乌鲁木齐830052

出　　处：《农业生物技术学报》2010年第1期149-155,共7页Journal of Agricultural Biotechnology

基　　金：新世纪人才支持计划(NCET-06-0108);国家"十一五"科技支撑计划(2007BAD36B02);国家自然科学基金(30871696)共同资助

摘　　要：为分离和鉴定新疆野苹果(Malus sieversii Roem.)根组织干旱胁迫条件下差异表达的基因,以苗龄为3个月的新源1号新疆野苹果水培苗为材料,应用20%PEG6000溶液处理模拟干旱胁迫,利用抑制性差减杂交(SSH)的方法构建了新疆野苹果根组织干旱胁迫诱导表达差减文库(正库)和抑制表达差减文库(反库)。正库和反库随机各挑菌500个,菌落PCR显示插入片段在250～1000bp之间。测序鉴定后共得到268个有效UniESTs。经Blastx和Blastn等生物信息学方法分析表明全部的UniESTs按可能的生物学功能可被分为16个大类,包括新陈代谢、能量、转录、蛋白质合成、蛋白质修饰降解、DNA加工、细胞信号转导等。To identify the genes controlling drought stress response in the root of Malus sieversii Roem.seedlings,suppression subtractive hybridization（SSH） was applied to construct forward and reverse subtractive cDNA libraries from M.sieversii Roem.Xinyuan 1 treated with 20% PEG 6000.After sequencings 500 clones respectively from each of two cDNA libraries at random,268 UniESTs were isolated.Homology search and sequences annotation were completed by Blastx and Blastn NCBI network service.According to MIPS function catalogue,all UniESTs were classified into 16 classifications.They were ascribed to proteins which were taken part in metabolism,transcription,protein biosynthesis and proteolytic,cell fate,cellular communication transduction,energymetabolismandsoon.

关 键 词：抑制性差减杂交 新疆野苹果 干旱胁迫 表达序列标签 

分 类 号：S661.1[农业科学—果树学]