芍药种子下胚轴休眠相关基因的cDNA-AFLP分析

doi:10.13560/j.cnki.biotech.bull.1985.2015.06.018

生物技术通报 ›› 2015, Vol. 31 ›› Issue (6): 116-121.doi: 10.13560/j.cnki.biotech.bull.1985.2015.06.018

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

收稿日期:2014-09-25 出版日期:2015-06-19 发布日期:2015-06-20
作者简介:孙晓梅，女，博士，教授，研究方向：园林植物遗传育种；E-mail：xiaomei7280@126.com
基金资助:
国家自然科学基金项目(31240028，31470696)

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

Received:2014-09-25 Published:2015-06-19 Online:2015-06-20

摘要/Abstract

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

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 metabolism etc. during the dormancy of seed hypocotyl of P. lactiflora. Nine TDFs were similar to genes of unknown functions and hypothetical proteins, and another 3 TDFs had no sequence homology to GenBank entries and might be new unknown genes, which could help us to clarify the mechanism of seed hypocotyl dormancy of P. lactiflora.

Key words: Paeonia lactiflora, seed, hypocotyl, dormancy, cDNA-AFLP, differentially expressed gene

孙晓梅, 韩宁宁, 杨宏光, 杨盼盼. 芍药种子下胚轴休眠相关基因的cDNA-AFLP分析[J]. 生物技术通报, 2015, 31(6): 116-121.

Sun Xiaomei, Han Ningning, Yang Hongguang, Yang Panpan. cDNA-AFLP Analysis of Dormancy-related Genes in Seed Hypocotyl of Paeonia lactiflora[J]. Biotechnology Bulletin, 2015, 31(6): 116-121.

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芍药种子下胚轴休眠相关基因的cDNA-AFLP分析

cDNA-AFLP Analysis of Dormancy-related Genes in Seed Hypocotyl of Paeonia lactiflora

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