灰毡毛忍冬转录组SSR位点分析及EST-SSR标记开发

Analysis on SSR Loci in Transcriptome and Development of EST-SSR Molecular Markers in Lonicera macranthoides

灰毡毛忍冬(Lonicera macranthoides)为中药山银花的一种基原植物,在中国南方广泛分布,具有很高的药用价值和经济价值。本研究利用MISA工具对灰毡毛忍冬转录组74057条Unigenes中的SSR位点进行了分析,其中15587条Unigene共计含有SSR位点20161个,SSR发生频率为21.05%。灰毡毛忍冬转录组SSR位点的种类从单核苷酸至六核苷酸重复均有分布,主要重复类型是二核苷酸(9704个)和三核苷酸(5847个),分别占SSR总数的48.13%和29.00%,主要重复单元为AG/CT(6086)、AT/AT(3071)和A/T(2826),分别占SSR总数的30.19%、15.23%和14.02%。SSR位点的重复次数分布最多的是6和5次,分别有4897和3531个,分别占总SSR数的24.29%和17.51%。利用Primer 6.0软件设计了17611对SSR引物,随机选出30对引物进行扩增验证,其中14对扩增条带表现出多态性,进而利用这14对引物对6个灰毡毛忍冬品种开展了遗传多样性分析。本研究结果将为灰毡毛忍冬物种鉴定、遗传多样性分析和分子标记辅助育种等提供科学依据。

As a species of original plant of flos Lonicerae,Lonicera macranthoides is widely distributed in South China and features high medicinal and economic value.In this study,the SSR loci of 74057 unigenes in the transcriptome of L.macranthoides were analyzed by MISA.Among them,15587 unigenes contained 20161 SSR loci,and the frequency of SSR was 21.05%.The types of SSR sites in the transcriptome of L.macranthoides were distributed from single nucleotide to six nucleotide repeats.The main repeats were dinucleotide(9704)and trinucleotide(5847),accounting for 48.13%and 29.00%of the total SSRs,respectively.The main repeat units were AG/CT(6086),AT/AT(3071)and A/T(2826),accounting for 30.19%,15.23%,and 14.02%of the total SSRs,respectively.There were 4897 and 3531 repeats(The numbers of repeats were the most)in 6 and 5 SSR loci,accounting for 24.29%and 17.51%of the total SSR number,respectively.By using primer 6.0 software,17611 pairs of SSR primers were designed,and 30 pairs of primers were randomly selected for amplification verification.Among them,14 pairs of amplified bands showed polymorphism,and then the genetic diversity of 6 varieties of L.macranthoides was analyzed by using these 14 pairs of primers.The results will provide a scientific basis for species identification,genetic diversity analysis,and molecular marker-assisted breeding of L.macranthoides.