李白盾蚧转录组分析及SSR位点开发  

Analysis of the transcriptome and development of SSR Loci in Pseudaulacaspis prunicola(Maskell)

作　　者：杜炫星 牛敏敏 赵清 蔡波 卢运运 魏久锋 张虎芳 DU Xuan-Xing;NIU Min-Min;ZHAO Qing;CAI Bo;LU Yun-Yun;WEI Jiu-Feng;ZHANG Hu-Fang(Shanxi Agricultural University,College of Plant Protection,Taigu 030801,Shanxi Province,China;Xinzhou Teachers University,Department of Biology,Xinzhou 034000,Shanxi Province,China;Haikou Customs,Tropical Plant Quarantine Center,Haikou 570311,China)

机构地区：[1]山西农业大学植物保护学院,山西太谷030801 [2]忻州师范学院生物系,山西忻州034000 [3]海口海关热带植物隔离检疫中心,海口570311

出　　处：《环境昆虫学报》2023年第5期1318-1332,共15页Journal of Environmental Entomology

基　　金：山西省归国留学人员科研资助项目(2020-065,2020-064);山西省基础研究计划(202103021224132,202103021224331);国家自然科学基金(32100370,31872272);山西农业大学创新基金项目(2020BQ79);海南省重点研发计划项目(ZDYF2018098)。

摘　　要：李白盾蚧Pseudaulacaspis prunicola(Maskell)寄主范围广泛,是一种重要的入侵害虫。本研究利用高通量测序平台(Illumina NovaSeq 6000)对李白盾蚧进行转录组测序、de novo从头组装及功能注释,在此基础上筛选其微卫星(SSR)位点,并挖掘微卫星引物。研究共获得李白盾蚧转录组60296条转录本,24967条单基因(unigenes)序列。通过GO数据库注释,将所有unigenes的功能分为生物学进程、细胞组分和分子功能三大类41个亚类功能区。KOG数据库注释结果显示,5085条unigenes归到25个基因家族,注释到一般功能预测的数目最多。KEGG代谢通路富集分析显示6668条unigenes注释到280个代谢通路,其中注释到内质网中蛋白质加工的数目最多。利用MISA软件共搜索到微卫星位点18193个,分布在9043条unigenes中,占总unigenes数量的36.22%,平均每2.29 kb出现一个SSR位点。其中主要重复类型为单核苷酸重复,占SSR位点总数的72.03%,其次为三核苷酸重复(15.90%)和二核苷酸重复(8.48%)。单核苷酸重复主要为A/T(71.16%),二核苷酸重复主要为AG/CT(5.20%)。基于Primer Primer 3软件设计出12538对李白盾蚧SSR引物,从中随机挑选50对引物进行PCR验证,共29对引物可以稳定扩增出目的片段。本研究成功组装了李白盾蚧转录组数据,并基于转录组数据成功筛选出其微卫星位点,为未来该虫的种群遗传学以及入侵生物学研究提供了数据支撑。Pseudaulacaspis prunicola(Maskell)is an important invasive pest with a wide host range.The transcriptome of P.prunicola was sequenced based on high-throughput sequencing platform(Illumina NovaSeq 6000),de novo assembled and function annotion;On the basis screened its microsatellite(SSR)loci and mined microsatellite primers.A total of 60296 transcripts and 24967 unigenes were obtained.According to Gene Ontology(GO)database annotation,these unigenes can be divided into 41 subclasses in 3 categories including biological process,cellularcomponent and molecular function.The results of KOG database annotation showed that 5085 sequences were classified into 25 gene families,with the most number annotated to the general function prediction.Enrichment analysis of KEGG metabolic pathways showed that 6668 unigenes were annotated to 280 metabolic pathways,andthe number of annotations to protein processing in endoplasmic reticulum was the largest.The microsatellite loci were searched using MISA software.A total of 18193 SSRs were distributed in 9043 unigene,with a frequency of 36.22%,with one SSR per 2.29 kb.The majority of microsatellite loci consisted of mono-,di-and tri-nucleotide motifs(72.03%,8.48% and 15.90%,respectively).The mono-nucleotide repeats motif were mainly A/T(71.16%)and the di-nucleotide repeats were mainly AG/CT(5.20%).Based on the 12538 pairs of SSR primers designed by Primer Primer 3 software,50 pairs of primers were randomly selected and 29 pairs were amplified successfully.In this study,we successfully assembled the transcriptome of P.prunicola and successfully developed its potential SSR makers based on P.prunicola transcriptome,will provide data support for future studies on the population genetics and invasion biology of this insect.

关 键 词：李白盾蚧 高通量测序 转录组分析 微卫星 分子标记 

分 类 号：Q963[生物学—昆虫学] S433[农业科学—农业昆虫与害虫防治]