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作 者 ：张 宇1，石凤翎1*，云 娜1，高 霞1，伊风艳1，蔡丽艳2

期 刊 ：西北植物学报 2013年 1期 页码：60~65

关键词：扁蓿豆；冷诱导；cDNA-AFLP；差异表达；

Keywords:Medicago ruthenica, cold-induced, cDNA-AFLP, different expression,

摘 要 ：以直立型扁蓿豆幼苗为试验材料，采用cDNA-AFLP技术分析扁蓿豆在低温胁迫诱导下的基因表达差异。结果显示，利用筛选的64对引物组合，对0 ℃低温处理3～5叶期扁蓿豆幼苗的叶片cDNA进行扩增，共获得549条差异表达的转录衍生片段（TDFs）。选取上调表达较好的43条片段进行克隆、测序、Blastx比对和功能分析，其中32个TDFs的蛋白序列与基础代谢、信号转导、转录因子、防御等功能有关，11个TDFs为假设蛋白、未知蛋白或没有找到一致序列。利用荧光定量PCR对3种不同上调表达差异片段进行验证，结果可从数值上更准确地显示差异片段在低温胁迫过程中的相对表达量。

Abstract:cDNA-AFLP approach was employed to analyze genes differential expression under cold stress using the three-five leaf stage leaves of Medicago ruthenica.The results showed that 549 differentially expressed transcript-derived fragments（TDFs） were identified by sixty-four primer combinations,and 43 up-regulated TDFs of them were sequenced and analyzed.Blastx-analysis and functional analysis indicated that 32 TDFs might participate in senescence and metabolism pathways,signal transduction,transcription regulation,stress responses etc,other 11 TDFs were showed homology to unknown and hypothetical function genes.The qRT-PCR results showed that it was accuately on the numerical value for expression of three different up-regulated TDFs were induced by low temperature stress.

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