蓝靛果忍冬转录组SSR信息分析及其分子标记开发

Analysis on SSR Information in Transcriptome and Development of Molecular Markers in Lonicera caerulea

利用MISA软件筛选蓝靛果忍冬(Lonicera caerulea L.)转录组测序获得的45 656条Unigene,共检测出14 841个SSR位点,分布于11 251条Unigene中,出现频率为32.51%,平均分布距离为2.58 kb。优势重复基序为单核苷酸、二核苷酸和三核苷酸,分别占总SSR位点的34.81%,42.79%和20.64%。二核苷酸重复基元中以AG/CT为优势重复基元,占总位点27.2%,三核苷酸重复基元以AAG/CTT为主,占6.18%。利用Primer 3.0共设计出21 867对SSR引物。随机选择20对引物进行PCR扩增,其中8对扩增出清晰可重复的条带,5对在16个蓝靛果忍冬材料中表现出多态性。利用UPGMA作图,将16份供试材料分为3类。丰富的SSR多态性为蓝靛果忍冬遗传多样性分析和遗传图谱构建提供更加丰富可靠的标记选择。

Forty-five thousand six hundred and fifty-six unigenes from fruit transcriptome of Lonicera caerulea L. were screened using MISA software. A total of 14 841 SSRs that occurred in 11 251 unigenes were identified,and the frequency of these SSRs was 32.51% and mean distance was 2.58 kb in the unigenes. Mononucleotide,dinucleotide and trinucleotide were major types,accounting for 34.81%,42.79% and 20.64%,respectively. The dinucleotide repeat motifs of AG/CT were the predominant repeat types（27.2%）. The trinucleotide repeat motifs of AAG/CTT were the predominant repeat types（6.18%）. Twenty-one thousand eight hundred and sixty-seven pairs of SSR primers were designed using Primer 3.0. Randomly 20 pairs of primers were selected for PCR amplification,8 amplified on clear and reproducible bands,5 in 16 different types showed polymorphism in materials. Sixteen plants were divided into 3 groups by UPGMA. Abundant SSR polymorphisms provide more reliable markers for Lonicera caerulea L. fruit genetic diversity analysis and genetic map construction.