河北师范大学学报—

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刊名：河北师范大学学报（自然科学版）Journal of Hebei Normal University (Natural Science)
主办：河北师范大学
ISSN： 1000-5854
CN： 13-1061/N
中国科技核心期刊
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中国高校优秀科技期刊
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基于转录组的褐软蜡蚧微卫星分子标记开发

(河北师范大学生命科学学院，河北石家庄 050024)
DOI： 10.13763/j.cnki.jhebnu.nse.202204011

Development of Microsatellite Markers Based on the Transcriptome-data of Coccus hesperidum（Hemiptera:Coccidae）

LI Dongzhi,ZHANG Meixia,SHAN Yiman,WANG Fang

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摘要/Abstract

摘要：

褐软蜡蚧Coccus hesperidum是一种在世界范围内危害多种经济作物和园林树木的昆虫. 基于高通量测序技术获得褐软蜡蚧的转录组数据，通过微卫量识别工具（MISA）筛选SSR位点，同时利用Primer Premier 3批量设计引物并进行验证. 结果显示，褐软蜡蚧的转录组数据经过拼接和聚类，最终获得111 073条单基因簇(unigenes)，长度为201~23 463 bp，平均长度为1 687 bp，并通过MISA预测到51 272个SSR位点. 其中，单核苷酸重复率最高（87. 14 %），三核苷酸重复率次之（8. 93 %），二核苷酸重复率最低（2. 74 %）；单核苷酸以A/T重复为主 (80. 16 %)，三核苷酸以CCG/CGG 重复为主(2. 33 %)，二核苷酸以AG/CT重复为主（1. 24 %）. 同时利用七大数据库进行功能注释，GO注释率最高，为47. 85 %. 此外，共随机挑选50对引物，其中15对引物在3个褐软蜡蚧地理种群中均成功扩增出目的条带. 通过转录组数据筛选褐软蜡蚧SSR位点，为褐软蜡蚧的种群遗传及功能基因等研究奠定基础.

Abstract：

Coccus hesperidum is an important economic pest causing harmful damage to many commercial crops and ornamental plants worldwide. The transcriptome data was obtained by the high-throughputsequencing, and the SSR loci were identified using MIcorSAtellite identification tool（MISA）, then the Primer Premier 3 was used to design primers for experimental validation. The results showed that the transcriptome of C. hesperidum include 111 073 unigenes, the length ranging from 201 bp to 23 463 bp, with an average length of 1 687 bp, and 51 272 SSR loci were predicted by MISA. Among them, the majority of these were mononucleotides (87. 14 %), followed by trinucleotides (8. 93 %) and dinucleotides (2. 74 %). The most abundant mononucleotide repeat motif was A/T (80. 16 %). The dominant motif among the tri-nucleotide repeat units was CCG/CGG (2. 33 %). The dominant motif among the dinucleotides repeat units was CCG/CGG (2. 33 %). And seven databases were used for functional annotation and the GO annotation was the most (47. 85 %). Additionally, out of 50 pairs of the randomly selected primers, a total of 15 primer pairs successfully amplificated the target bands of C. hesperidum from three different geographic populations. In short, 51 272 SSR loci were identified based on the transcriptome data, which can provide a foundation for research on molecular markers, population genetics and functional genes for C. hesperidu.

关键词

关键词： 褐软蜡蚧；转录组；微卫星DNA；分子标记

Key words： Coccus hesperidum ; transcriptome ; microsatellite DNA ; molecular marker

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