桃花芽休眠解除SSH文库构建及相关基因表达分析  

Construction of the Suppression Subtractive Hybridization Library and Analysis of Related Genes of Floral Buds in Prunus persica During Dormancy-releasing

作　　者：李玲[1] 王慧[1] 谭钺[1] 王宇[1] 陈修德[1] 李冬梅[1] 高东升[1] 

机构地区：[1]山东农业大学园艺科学与工程学院,作物生物学国家重点实验室,山东泰安271018

出　　处：《园艺学报》2011年第12期2273-2280,共8页Acta Horticulturae Sinica

基　　金：‘十二五’国家科技支撑计划项目(2011BAD12B02);山东省现代农业产业技术体系、水果创新团队专项

摘　　要：为了深入研究桃花芽休眠的分子机制,利用抑制差减杂交技术(SSH),以解除休眠期的花芽的RNA为实验方Tester,以休眠期花芽的RNA为驱动方Driver,构建休眠解除cDNA文库,测定分析了180个上调表达的cDNA片段,测序成功128个,除去冗余序列,得到28个单一序列。对序列进行BLAST比对及功能注释,发现这些序列分别属于新陈代谢、氧化还原、信号转导、抗性相关等类别基因。运用qRT-PCR技术对HisH3、Dhn和Metallothionein基因进行检测,结果表明它们在花芽休眠解除过程中均上调表达。To better understand the molecular and physiological mechanisms of seasonal bud dormancy in perennial trees,the differentially expressed genes in peach[Prunus persica（L.）Batsch]floral buds during dormancy-releasing were identified by suppression subtractive hybridization（SSH）.One hundred and eighty positive clones were selected and sequenced from the libraries totally,and 28 unique genes were obtained.According to the BLAST screening and functional annotation,metabolism,oxidation reduction,signal transduction and defense-related genes were identified with high frequencies in fertility cDNA library.In SSH library three representative genes,HisH3,Dhn and Metallothionein were selected and analyzed qRT-PCR.The results showed that they were up-regulated during dormancy-releasing of floral buds.

关 键 词：桃 花芽 休眠 抑制差减杂交 基因表达 QRT-PCR 

分 类 号：S662.1[农业科学—果树学]