ABiMS Bioinformatics Facility, CNRS, Sorbonne Université, FR2424, Station Biologique, 29680, Roscoff, France.
UMR PEGASE, INRAE, Institut Agro, 16 Le Clos, 35590, Saint-Gilles, France.
BMC Genomics. 2020 Oct 2;21(1):687. doi: 10.1186/s12864-020-07099-4.
Common Pekin and Muscovy ducks and their intergeneric hinny and mule hybrids have different abilities for fatty liver production. RNA-Seq analyses from the liver of these different genetic types fed ad libitum or overfed would help to identify genes with different response to overfeeding between them. However RNA-seq analyses from different species and comparison is challenging. The goal of this study was develop a relevant strategy for transcriptome analysis and comparison between different species.
Transcriptomes were first assembled with a reference-based approach. Important mapping biases were observed when heterologous mapping were conducted on common duck reference genome, suggesting that this reference-based strategy was not suited to compare the four different genetic types. De novo transcriptome assemblies were then performed using Trinity and Oases. Assemblies of transcriptomes were not relevant when more than a single genetic type was considered. Finally, single genetic type transcriptomes were assembled with DRAP in a mega-transcriptome. No bias was observed when reads from the different genetic types were mapped on this mega-transcriptome and differences in gene expression between the four genetic types could be identified.
Analyses using both reference-based and de novo transcriptome assemblies point out a good performance of the de novo approach for the analysis of gene expression in different species. It also allowed the identification of differences in responses to overfeeding between Pekin and Muscovy ducks and hinny and mule hybrids.
普通北京鸭和麝香鸭及其种间杂种骡和驴在生产脂肪肝方面的能力不同。对这些不同遗传类型的肝脏进行无限制或过度喂养的 RNA-Seq 分析将有助于识别对过度喂养反应不同的基因。然而,来自不同物种的 RNA-seq 分析和比较具有挑战性。本研究的目的是开发一种相关的转录组分析策略,并在不同物种之间进行比较。
首先使用基于参考的方法对转录组进行组装。当在普通鸭参考基因组上进行异源映射时,观察到重要的映射偏差,这表明这种基于参考的策略不适合比较四种不同的遗传类型。然后使用 Trinity 和 Oases 进行从头转录组组装。当考虑多个遗传类型时,转录组的组装就不相关了。最后,使用 DRAP 在 mega-transcriptome 中组装单个遗传类型的转录组。当将来自不同遗传类型的reads映射到此 mega-transcriptome 时,没有观察到偏差,并且可以识别出四种遗传类型之间基因表达的差异。
基于参考和从头转录组组装的分析都指出了从头转录组组装方法在分析不同物种中的基因表达方面的良好性能。它还允许识别出北京鸭和麝香鸭以及骡和驴杂种之间对过度喂养的反应差异。