• 中文核心期刊
  • CSCD来源期刊
  • 中国科技核心期刊
  • CA、CABI、ZR收录期刊

利用芥菜转录组信息挖掘SSR标记及用于种质分析

Using SSR Markers from Brassica juncea Transcriptome for Germplasm Analysis

  • 摘要:
      目的  为开发高效的芥菜Brassicajuncea.分子标记。
      方法  本研究通过对芥菜转录组测序的信息搜索SSR位点并分析其分布特点,应用Primer 3.0软件设计SSR引物,随机选择 50 对引物扩增44 份芥菜种质,检测其多态性。
      结果  芥菜转录组测序共获得55 636条unigene,全部序列有48 193 376 bp;有7 834条unigene包含SSR的序列,从中鉴定出9 526个SSR位点,其中有1 371条序列包含1个以上SSR,复合SSR有572个,SSR发生频率为14.08%。优势重复基序为三核苷酸和二核苷酸,分别占总SSR的51.12%和41.91%。二核苷酸重复基元中以AG/CT为优势重复基元,占总位点的34.74%,三核苷酸重复基元以AAG/CTT为主,占总位点的18.30%。共设计出21 282对SSR引物,随机选择 50 对引物进行 PCR 扩增,其中41对扩增出清晰可重复的预期条带,17对(占34%)在 44 份芥菜种质中表现出多态性。应用UPGMA得到将44份供试材料分为4大类聚类图,可准确地体现了芥菜种质材料的关系。
      结论  根据芥菜转录组数据能开发出类型丰富、效率较高的SSR标记,为芥菜亲缘关系分析和遗传图谱构建等提供更可靠的标记。

     

    Abstract:
      Objective  To search for applicable molecular markers for studying Brassica juncea.
      Method  SSR sites were searched from transcriptional sequencing of B. juncea. Potential primers were designed using Primer 3.0, and polymorphisms of 44 B. juncea germplasms detected from randomly selected 50 pairs of primers for the investigation.
      Result  Totally, 55,636 unigenes were obtained from the transcriptome sequencing. The entire sequence was 48,193,376bp with 9,526 SSR loci identified from 7,834 unigenes at a rate of 14.08%. Of the loci, 1,371 contained more than one SSR locus and 572 compound loci. Of the major repeat types, 51.12% were trinucleotide dominated by AGA/TCT (18.30%), and 41.91% dinucleotide dominated by AG/CT (34.74%). Using Primer 3.0, from 21,282 pairs of SSR primers, 50 were randomly selected for the PCR amplification. Out of 44 amplifications, 41 showed clear and reproducible bands, and 17 with polymorphisms. The 44 germplasms were classified into 4 distinctive groups by UPGMA.
      Conclusion  The SSR markers with high frequency and polymorphism could be acquired from the transcriptome to be used for the genetic diversity analysis and mapping construction on B. juncea.

     

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