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[目的]开发银鲳分子标记技术。[方法]通过对银鲳(Pampus argenteus)进行高通量转录组测序(RNA-seq)获得银鲳转录组原始试验数据,经过拼接后,获得3 715 603条unigene序列。采用生物信息学分析软件MISA对所有unigene进行简单重复序列(simple sequence repeat,SSR)位点鉴定。同时,利用软件对银鲳转录组SSR的多态性进行评价。[结果]银鲳转录组水平上,共鉴定出107 007个SSR位点,分布在97 289条unigene中,发生频率为2.62%,SSR平均密度为476个/Mbp。在银鲳转录组SSRs中,单核苷酸与二核苷酸重复序列为主要重复类型,分别占总SSRs的48.14%和34.10%。银鲳转录组数据中SSR序列共包括424种重复基元类型,单核苷酸重复基元A占较高比例,占同一重复类型SSRs的49.57%,二核苷酸重复基元TG/AC和三核苷酸重复基元GAG/AAC是优势重复基元,分别占同一重复类型SSRs的42.43%和9.61%。重复序列长度在12 bp以上的SSR标记位点数占总SSR的76.95%,具有丰富的多态性。[结论]银鲳SSRs位点具有极大的可开发性,可为银鲳遗传多样性研究、遗传图谱构建及分子辅助育种提供有效工具。
[Objective] The research aimed to develop silver-labeled molecular marker technology. [Method] The original experimental data of Agrobacterium rhizogenes transcriptome was obtained by high-throughput transcriptome sequencing (RNA-seq) of Pampus argenteus. After splicing, 3 715 603 unigene sequences were obtained. Bioinformatics analysis software MISA was used to identify all unigene simple sequence repeat (SSR) loci. At the same time, the software was used to evaluate the polymorphism of SSR in transcriptome. [Result] A total of 107 007 SSR loci were found in 97 289 unigene transcriptomes, with a frequency of 2.62% and an average SSR of 476 loci / Mbp. In SSRs, the single nucleotide and dinucleotide repeats were the main repeat types, accounting for 48.14% and 34.10% of the total SSRs, respectively. A total of 424 repeat haplotypes were included in the SSR sequences of the Agkistrodon acutus transcriptome data. The mononucleotide repeat A accounted for 49.57% of the same repeat SSRs, dinucleotide repeat motif TG / AC and Trinucleotide repeat motif GAG / AAC is the dominant repeat motif, accounting for 42.43% and 9.61% of the same repeat type SSRs, respectively. The number of SSR markers with repeat length over 12 bp accounted for 76.95% of the total SSR, which was rich in polymorphism. [Conclusion] The SSRs loci of Agkistrodondellus were highly exploitable and could provide useful tools for the genetic diversity, genetic map building and molecular assisted breeding of Agkistrodon.