基于转录组序列的乌桕SSR分子标记开发

Development of SSR Markers Based on Transcriptome Sequences of Triadica sebifera

为了开发乌桕(Triadica sebifera(L.)Small)的简单重复序列(SSR)分子标记,利用Illumina HiSeq2500测序平台对乌桕嫩叶进行转录组测序,基于测序获得的59441条Unigenes数据,利用MISA软件进行SSR位点鉴别,通过普通PCR和毛细管电泳筛选稳定可靠的多态性SSR标记。结果表明:从乌桕59441条Unigenes中鉴定出7344个包含1~6个核苷酸重复类型的SSR位点,出现频率为12.36%;单核苷酸、三核苷酸和二核苷酸为优势重复类型,其数量和占比分别为2946(40.11%)、2314(31.53%)和1958(21.87%)。从120对随机选择的SSR引物中筛选出了10对扩增效率高、稳定性好的多态性SSR引物。本研究为乌桕现存资源的遗传多样性研究、遗传图谱构建提供了有价值的候选标记。

To develop SSR markers based on transcriptome data of Triadica sebifera(L.)Small,the transcriptome of T.sebifera young leaves was sequenced by Illumina HiSeq 2500 platform.SSR loci were identified and analysed in all of 59441 Unigenes by using MISA software.Stable reliable polymorphic SSR markers were selected by using conventional PCR and capillary electrophoresis.The results showed that a total of 7344 SSR loci,including the types of 1~6 nucleotide repeats with occurring frequency of 12.36%,were identified from 59441 Unigenes in T.sebifera transcriptome.Mononucleotide,trinucleotide and dinucleotide repeats were the dominant types,accounted for as much as 2946(40.11%),2314(31.53%),and 1958(21.87%)of all SSRs,respectively.A total of 120 pairs of SSR primers were randomly selected,among which 10 were verified to be polymorphic primers.This study can provide valuable candidate markers for genetic diversity analysis and genetic mapping construction of existing T.sebifera resource.