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目的:通过对灯盏花转录组中SSR位点信息的分析,设计SSR引物,为开发新的SSR标记奠定了基础。方法:利用MISA工具筛选灯盏花转录组测序获得的52 060条unigenes,对其SSR位点信息进行了分析;Primer3设计SSR引物,随机选取36对引物对13株采自不同地方的灯盏花进行多态性扩增分析。结果:灯盏花转录组搜索到3 639个SSR位点,分布于3 260条unigenes上,SSR位点出现频率是6.99%。其中,二核苷酸重复是主要的类型,占总SSR的34.41%,其次是单核苷酸和三核苷酸重复基元,分别占总SSR的31.41%,30.04%。二核苷酸重复基元中以AT/AT和AC/GT为优势重复基元,占总SSRs的28.71%,三核苷酸重复基元以AAT/ATT为主,占7.94%。随机选取36对引物进行PCR扩增,其中34对(94.44%)扩增出清晰、可重复的条带,19对(52.78%)表现出多态性差异。利用UPGMA作图,将13个材料分为2类。结论:灯盏花转录组中SSR位点出现频率高,类型丰富;大量的SSR为其遗传多样性分析和遗传图谱构建提供了丰富的候选分子标记。
OBJECTIVE: To design SSR primers based on the analysis of SSR loci in the transcriptome of Breviscapus, which laid the foundation for the development of new SSR markers. Methods: 52 060 unigenes obtained from the transcriptome of Breviscapus were screened by MISA tool to analyze the SSR loci. Primer3 was designed with SSR primers and randomly selected 36 pairs of primers for 13 strains of Breviscapus collected from different places Amplified amplification analysis. RESULTS: The 3 639 SSR loci were found in the transcriptome of Breviscapus, which distributed on 3 260 unigenes and the frequency of SSR loci was 6.99%. Among them, dinucleotide repeats were the major types, accounting for 34.41% of the total SSRs, followed by mononucleotide and trinucleotide repeats, accounting for 31.41% and 30.04% of the total SSR, respectively. The dinucleotide repeat motifs were dominated by AT / AT and AC / GT, accounting for 28.71% of the total SSRs. The trinucleotide repeat motifs were mainly AAT / ATT, accounting for 7.94%. 36 pairs of primers were randomly selected for PCR amplification, of which 34 pairs (94.44%) amplified clear and repeatable bands, and 19 pairs (52.78%) showed polymorphic differences. Using UPGMA mapping, the 13 materials were divided into two categories. Conclusion: The frequency of SSR loci is high and the types are abundant in the transcriptome of Erigeron breviscapus. A large number of SSRs provide abundant candidate molecular markers for genetic diversity analysis and genetic map construction.