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利用抑制消减杂交技术构建东农冬麦1号分蘖节低温胁迫相关基因的cDNA正反文库,获得表达序列标签EST(Expressed sequence tag),经分析和前期处理后拼接得到736条非重复序列(Unigenes)。运用生物信息学方法对低温胁迫下诱导的冬小麦东农冬麦1号分蘖节相关EST-SSRs分布频率和碱基重复特点进行归纳分析。结果表明,736条Unigenes共检索到59个EST-SSRs,EST-SSRs出现频率为8.02%。EST-SSRs包括24种重复单元,其主要以三核苷酸和二核苷酸重复基元为主,二者分别占总SSRs 47.46%和44.07%,在EST中出现频率分别为3.80%和3.53%,其中以二核苷酸CA和AG重复基元为主,三核苷酸重复基元以AAC重复基元所占比例最高。为冬小麦EST资源开发和抗寒基因挖掘奠定基础。
Using suppression subtractive hybridization (SSH) technique, a cDNA positive and negative library of Dongnongdongmai tillering stage cold stress-related genes was constructed and expressed sequence tags EST (Expressed sequence tag) was obtained. After analysis and pre-treatment, 736 non-repetitive sequences ). Bioinformatics method was used to analyze the distribution frequency and base repetition characteristics of EST-SSRs related to tillering stage of Dongnong Dongmai1 induced by low temperature stress. The results showed that 59 EST-SSRs were found in 736 Unigenes, and the frequency of EST-SSRs was 8.02%. The EST-SSRs consisted of 24 kinds of repeat units, which were mainly trinucleotide and dinucleotide repeat motifs, which accounted for 47.46% and 44.07% of the total SSRs, respectively, and the ESTs were found in ESTs with frequency of 3.80% and 3.53 %, Of which the dinucleotide CA and AG repeat motifs are the main trinucleotide repeat motifs with the highest proportion of AAC repeat motifs. It laid the foundation for winter wheat EST resource development and cold hardiness gene mining.