芍药种子下胚轴休眠相关基因的cDNA-AFLP分析.pdf

生物技术通报 ·研究报告· BIOTECHNOLOGY BULLETIN ( ) 2015, 31 6 :116-121 芍药种子下胚轴休眠相关基因的 cDNA-AFLP 分析 孙晓梅  韩宁宁  杨宏光  杨盼盼 （沈阳农业大学林学院，沈阳 110866） 摘 要 ： 旨在探讨芍药种子下胚轴的休眠机理，采用cDNA-AFLP 技术对层积0 d 和根部露白两个时期的芍药种子进行基因 差异表达分析。利用256 对引物组合进行扩增，筛选获得3 600 个差异表达转录衍生片段 （TDFs），成功回收到1 200 个TDFs，选 取500 个TDFs 对其克隆测序，共得到42 个有效序列。经BLAST 比对发现，其中30 个TDFs 与已知功能基因同源，分别参与芍药 种子下胚轴休眠过程中的基因表达调控、信号转导、逆境胁迫、物质与能量代谢等过程，9 个TDFs 与功能未知基因及假想蛋白同源， 另有3 个TDFs 为无同源序列，可能是新的未知基因，这些基因有助于更好地阐明芍药种子下胚轴休眠机理。 关键词 ： 芍药；种子 ；下胚轴 ；休眠 ；cDNA-AFLP ；差异表达基因 DOI ：10.13560/ki.biotech.bull.1985.2015.06.018 - - cDNA AFLP Analysis of Dormancy related Genes in Seed Hypocotyl of Paeonia lactiflora Sun Xiaomei Han Ningning Yang Hongguang Yang Panpan （College of Forestry ，Shenyang Agricultural University ，Shenyang 110866 ） Abstract: To explore the dormancy mechanism of seed hypocotyl of Paeonia lactiflora and isolate dormancy-related genes from P. lactiflora, a cDNA-AFLP technology was used to identify differentially expressed genes between the stages of seeds with stratification for 0 d and germination. A total of 3 600 differentially expressed transcript-derived fragments（TDFs）were screened by selective PCR amplification using 256 primer combinations, and 1 200 TDFs were recovered successfully. Then selected 500 TDFs were cloned and sequenced, and 42 effective sequences were obtained. Bioinformatics analysis with BLAST showed that 30 TDFs were homologous to the known functional genes, which involved in gene expression regulation, signal transduction, stress responding, material and energy metabo