Roulin Anne C, Wu Min, Pichon Samuel, Arbore Roberto, Kühn-Bühlmann Simone, Kölliker Mathias, Walser Jean-Claude
Department of Environmental Sciences, Zoology and Evolution, University of Basel, Basel, Switzerland.
Department of Environmental Sciences, Zoology and Evolution, University of Basel, Basel, Switzerland; Genetic Diversity Centre (GDC), ETH Zürich, Zürich, Switzerland.
PLoS One. 2014 Apr 10;9(4):e94098. doi: 10.1371/journal.pone.0094098. eCollection 2014.
The European earwig (Forficula auricularia) is an established system for studies of sexual selection, social interactions and the evolution of parental care. Despite its scientific interest, little knowledge exists about the species at the genomic level, limiting the scope of molecular studies and expression analyses of genes of interest. To overcome these limitations, we sequenced and validated the transcriptome of the European earwig.
To obtain a comprehensive transcriptome, we sequenced mRNA from various tissues and developmental stages of female and male earwigs using Roche 454 pyrosequencing and Illumina HiSeq. The reads were de novo assembled independently and screened for possible microbial contamination and repeated elements. The remaining contigs were combined into a hybrid assembly and clustered to reduce redundancy. A comparison with the eukaryotic core gene dataset indicates that we sequenced a substantial part of the earwig transcriptome with a low level of fragmentation. In addition, a comparative analysis revealed that more than 8,800 contigs of the hybrid assembly show significant similarity to insect-specific proteins and those were assigned for Gene Ontology terms. Finally, we established a quantitative PCR test for expression stability using commonly used housekeeping genes and applied the method to five homologs of known sex-biased genes of the honeybee. The qPCR pilot study confirmed sex specific expression and also revealed significant expression differences between the brain and antenna tissue samples.
By employing two different sequencing approaches and including samples obtained from different tissues, developmental stages, and sexes, we were able to assemble a comprehensive transcriptome of F. auricularia. The transcriptome presented here offers new opportunities to study the molecular bases and evolution of parental care and sociality in arthropods.
欧洲蠼螋(Forficula auricularia)是用于性选择、社会互动及亲代抚育进化研究的成熟系统。尽管其具有科学研究价值,但在基因组水平上对该物种的了解甚少,这限制了分子研究及相关基因表达分析的范围。为克服这些限制,我们对欧洲蠼螋的转录组进行了测序和验证。
为获得全面的转录组,我们使用罗氏454焦磷酸测序和Illumina HiSeq对雌性和雄性蠼螋不同组织及发育阶段的mRNA进行了测序。读取的序列进行了独立的从头组装,并筛选了可能的微生物污染和重复元件。剩余的重叠群被合并成一个混合组装并聚类以减少冗余。与真核核心基因数据集的比较表明,我们测序了蠼螋转录组的大部分,且片段化程度较低。此外,比较分析显示,混合组装的8800多个重叠群与昆虫特异性蛋白具有显著相似性,并被赋予了基因本体论术语。最后,我们使用常用的看家基因建立了表达稳定性的定量PCR检测方法,并将该方法应用于蜜蜂已知性别偏向基因的五个同源基因。qPCR初步研究证实了性别特异性表达,还揭示了脑和触角组织样本之间的显著表达差异。
通过采用两种不同的测序方法,并纳入来自不同组织、发育阶段和性别的样本,我们成功组装了欧洲蠼螋的全面转录组。这里呈现的转录组为研究节肢动物亲代抚育和社会性的分子基础及进化提供了新机会。