基于转录组数据高通量发掘扶桑绵粉蚧微卫星引物

High-throughput discovery of SSR genetic markers in the mealybug,Phenacoccus solenopsis( Hemiptera: Pseudococcidae ), from its transcriptome database

【目的】扶桑绵粉蚧Phenacoccus solenopsis Tinsley是我国重要的检疫性害虫。简单重复序列(simple sequence repeat,SSR)研究在遗传图谱和物理图谱的构建、分子标记辅助育种、品种鉴定、基因定位、遗传多样性、动植物分类和进化等方面具有重要意义。筛选的SSR引物将为扶桑绵粉蚧遗传多样性分析、进化分析及入侵生物学等奠定基础。【方法】利用高通量搜索的方法对扶桑绵粉蚧转录组中28 120条unigenes的数据进行搜索。【结果】共找到1 781个SSR位点。扶桑绵粉蚧转录组中SSRs的主要重复类型是单核苷酸重复,占SSR总数的89.44%;其次是三核苷酸重复,占SSR总数的7.52%。单核苷酸重复里主要是A/T基序,占了总量的87.42%。基于筛选的SSRs,运用Primer 3软件进行引物的批量设计,共有481个unigenes成功设计引物,共设计出1 228对引物。【结论】研究表明利用扶桑绵粉蚧转录组数据开发SSR标记是可行的,本研究开发的引物将为扶桑绵粉蚧遗传多样性分析、进化分析及入侵生物学等奠定基础。

[ Aim ] Phenacoccus solenopsis Tinsley is one of the most important quarantine pests in China. Simple sequence repeats （SSRs） are very useful molecular markers in genetic studies such as genetic mapping, physical mapping, marker-assisted breeding and species identification, gene location, genetic diversity, classification and evolution of plants and animals, etc. Screening SSR primers will provide an important resource for marker-assisted investigation in genetic diversity, phylogenetic analysis and biological invasion of P. solenopsis. [ Methods ] A high-throughput method was used to develop the SSR genetic markers from the 28 120 unigenes of the transcriptome in the mealybug P. solenopsis. [ Results] In total, 1 781 SSR genetic markers were developed from the previously constructed transcriptome database. The majority of them contained mono- and trinucleotide motifs （89.44% and 7.52%, respectively）, of which A/T was the most abundant mononucleotide motif （87.42%）. Based on the identified SSRs, 1 228 pairs of primer sets were designed from 481 unigenes by the Primer 3 program. [ Conclusion ] Our study shows that it is feasible to develop SSR markers by using P. solenopsis transcriptome, and the primers developed in this study provide the foundation for the analysis of genetic diversity, evolutionary biology, and invasion biology of P. solenopsis.