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Model of gene expression in extreme cold - reference transcriptome for the high-Antarctic cryopelagic notothenioid fish Pagothenia borchgrevinki

机译:高南极低温拟南芥类鱼Pagothenia borchgrevinki的极寒参考转录组中的基因表达模型。

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Background Among the cold-adapted Antarctic notothenioid fishes, the high-latitude bald notothen Pagothenia borchgrevinki is particularly notable as the sole cryopelagic species, exploiting the coldest and iciest waters of the Southern Ocean. Because P. borchgrevinki is a frequent model for investigating notothenioid cold-adaptation and specialization, it is imperative that “omic” tools be developed for this species. In the absence of a sequenced genome, a well annotated reference transcriptome of the bald notothen will serve as a model of gene expression in the coldest and harshest of all polar marine environments, useful for future comparative studies of cold adaptation and thermal responses in polar teleosts and ectotherms. Results We sequenced and annotated a reference transcriptome for P. borchgrevinki, with added attention to capturing the transcriptional responses to acute and chronic heat exposures. We sequenced by Roche 454 a normalized cDNA library constructed from pooled mRNA encompassing multiple tissues taken from environmental, warm acclimating, and acute heat stressed specimens. The resulting reads were assembled into 42,620 contigs, 17,951 of which could be annotated. We utilized this annotated portion of the reference transcriptome to map short Illumina reads sequenced from the gill and liver of environmental specimens, and also compared the gene expression profiles of these two tissue transcriptomes with those from the temperate model fish Danio rerio. From this, we identified a conserved group of 58 GO terms, in which terms related to transcription and its regulation, ubiquitin-protein ligase activity, protein ubiquitination, and protein binding among others are more prevalent in the bald notothen, suggesting the pertinent genes play essential roles in cold temperature functioning. Conclusion We sequenced multiple tissue transcriptomes from native and heat-exposed experimental specimens of the high Antarctic, cryopelagic notothenioid P. borchgrevinki to construct a reference transcriptome. In a proof of concept, we utilized the annotated reference transcriptome to profile the gene expression patterns of gill and liver, and identified a suite of over and under-represented GO terms when compared to the tropical water zebrafish suggesting these functions may be important for surviving in freezing waters. The transcriptome resource from this study will aid future investigations of cold adaptation and thermal response of polar ectothermic species.
机译:背景技术在适应寒冷的南极类胡萝卜素鱼类中,高纬度秃头的齿形紫檀Pagothenia borchgrevinki是唯一的低温古生物种,它利用了南洋最冷和最冰冷的海水。由于P. borchgrevinki是调查类胡萝卜素冷适应性和特化性的常用模型,因此必须为该物种开发“组学”工具。在没有测序基因组的情况下,光秃的牙本质的参考转录组将被很好地注释,它将成为所有极地海洋环境中最冷和最严酷的环境中基因表达的模型,可用于将来对极地硬骨鱼的冷适应性和热响应进行比较研究和等温线。结果我们对P. borchgrevinki的参考转录组进行了测序和注释,并特别注意捕获对急性和慢性热暴露的转录反应。我们通过Roche 454对标准化的cDNA文库进行了测序,该文库由合并的mRNA构建而成,该mRNA涵盖了从环境,温暖适应和急性热应激标本中采集的多个组织。所得的读段被组装成42620个重叠群,其中17951个可以被注释。我们利用参考转录组的这个带注释的部分来绘制从环境标本的g和肝脏测序的短的Illumina读图,并且还将这两个组织转录组的基因表达谱与温带模型鱼Danio rerio的基因表达谱进行了比较。由此,我们确定了一个由58个GO术语组成的保守组,其中与转录及其调控,泛素蛋白连接酶活性,蛋白泛素化以及蛋白结合等相关的术语在秃头牙髓中更为普遍,表明相关基因发挥了作用。低温功能中的重要作用。结论我们对来自南极,低温的类胸腺类拟南芥P. borchgrevinki的天然和热暴露实验标本的多个组织转录组进行了测序,以构建参考转录组。在概念验证中,我们利用带注释的参考转录组分析了ill和肝脏的基因表达模式,并与热带水斑马鱼进行了比较,发现了一组过高和过低的GO术语,表明这些功能对于生存至关重要在冰冻的水中。这项研究的转录组资源将有助于未来对极地物种的冷适应和热响应的研究。

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