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作 者 ：孙宏冉;齐凤慧;詹亚光*;叶鹏

期 刊 ：植物研究 2014年 34卷 03期 页码：393-402

关键词：水曲柳；简单重复序列；EST-SSR；

Keywords:Fraxinus mandshurica Rupr., simple sequence repeat, EST-SSR,

摘 要 ：为使EST-SSRs分子标记技术在水曲柳辅助育种、基因定位、雌雄鉴定等研究中应用，本文对NCBI数据库中的12 100条水曲柳ESTs进行了分析，序列拼接得到全长为3.02×10⁶ bp的非冗余的5 423个ESTs聚类，利用在线软件SSRIT进行SSRs位点搜索，共检索出9 489个SSRs位点。EST-SSRs的长度从6~46 bp之间变化。二核苷酸重复在水曲柳中占主导地位(6 914个)，占总SSRs的72.86%，其次是三核苷酸(24.96%）和四核苷酸(1.37%)。各种不同重复模式中出现最多的是(AG/CT)n(3 458次)，占二核苷酸重复的50.01%，其次分别是(AC/GT)n(24.47%)和(AT/AT)n(24.23%)。利用Primer 5.0软件共设计了147对EST-SSRs引物。水曲柳ESTs中SSRs出现频率较高，类型丰富，这一研究结果提供了水曲柳EST-SSRs信息，使该分子标记技术在水曲柳研究中的进一步应用成为可能。

Abstract:In order to apply the EST-SSRs technique in molecular marker aided breeding, gene localization, and gender identification of Fraxinus mandshurica Rupr., a total of 12 100 ESTs from NCBI database were analyzed. The sizes of non redundant ESTs added up to 3.02×10⁶ bp, which can be clustered into 5 423 clads. A total of 9 489 SSRs loci (6-46 bp in length) were found via SSRIT software. Dinucleotide repeats is a predominated feature of these loci (accounting 72.86%), followed by trinucleotide repeats (24.96%) and tetranucleotide repeats (1.37%). The pattern (AG/CT)n is the most frequent one among all repeat patterns, followed by (AC/GT)n and (AT/AT)n. We designed 147 pairs of EST-SSRs primers by Primer 5.0 software. EST-SSRs in F.mandshurica Rupr displayed a relatively high level of occurrence frequency with abundant patterns. These results will pave the way for application of EST-SSRs molecular marker in F.mandshurica breeding.

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