Abstract: Gynaephora qinghaiensis is one of the important pests in the Qinghai-Tibet Plateau. In this study,transcriptome sequencing was performed on G.qinghaiensis adults and 4th instar larvae using the Illumina HiSeq^TM2000 high-throughput sequencing platform. On this basis,its microsatellite (SSR) locus was screened and the microsatellite primers were mined. 63 335 unigenes were obtained with 12 597 microsatellite loci distributed in 9 851 unigenes. Analysis of KOG,GO annotation and KEGG pathway database revealed that unigenes annotation mainly focused on general function prediction,cell progression and carbohydrate metabolism processes. SSR repeat types were mainly mononucleotide and dinucleotide repeats,(A/T) _n was the most dominant motif in mononucleotide repeats,accounting for 71.37% of the total SSR sites;(AC/GT) _n was the dominant motif of dinucleotide repeats,accounting for 6.06% of the total SSR sites. The number of SSR decreased with the number of repeats and the repeat type decreases with the length of the base sequence. Using Primer 3 software,2 714 SSR primers were designed,25 pairs were randomly selected for validation,and 11 pairs amplified the target DNA fragment. In this study,their microsatellite loci were successfully screened based on transcriptomic data,providing data support for further studies of their population genetics and occurrence dynamics.

Key words: Gynaephora qinghaiensis, Transcriptome analysis, Microsatellite loci, Molecular markers

摘要： 青海草原毛虫(Gynaephora qinghaiensis)是青藏高原牧区重要的害虫之一。本研究采用Illumina HiSeq^TM2000高通量测序平台对青海草原毛虫成虫和4龄幼虫进行转录组测序,在此基础上筛选其微卫星(Single sequence reperts,SSR)位点并挖掘微卫星引物。本研究共获得63 335条unigenes,有12 597个微卫星位点分布于9 851条unigenes中。通过KOG,GO注释和KEGG通路数据库分析发现,unigenes注释主要集中于一般功能预测、细胞进程和碳水化合物代谢过程。SSR重复类型主要为单核苷酸和二核苷酸重复,(A/T)_n是单核苷酸重复中最主要的基元类型,占总SSR位点的71.37%,(AC/GT)_n为二核苷酸重复的优势基元,占总SSR位点数的6.06%,且SSR数量随重复次数增加而降低,重复次数类型随基元序列长度增长而减少。利用Primer 3软件设计出2 714对青海草原毛虫SSR引物,随机挑出25对引物进行PCR验证,有11对引物扩增出目的DNA片段。本研究基于转录组数据成功筛选出青海草原毛虫微卫星位点,为进一步研究其种群遗传学和发生动态提供数据支撑。

关键词: 青海草原毛虫, 转录组分析, 微卫星, 分子标记