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对北大西洋主要浮游动物——哲水蚤(甲壳纲,哲水蚤目)进行转录组的从头组装

De novo assembly of a transcriptome for Calanus finmarchicus (Crustacea, Copepoda)--the dominant zooplankter of the North Atlantic Ocean.

作者信息

Lenz Petra H, Roncalli Vittoria, Hassett R Patrick, Wu Le-Shin, Cieslak Matthew C, Hartline Daniel K, Christie Andrew E

机构信息

Békésy Laboratory of Neurobiology, Pacific Biosciences Research Center, University of Hawaii at Manoa, Honolulu, Hawaii, United States of America.

Department of Biological Sciences, Ohio University, Athens, Ohio, United States of America.

出版信息

PLoS One. 2014 Feb 19;9(2):e88589. doi: 10.1371/journal.pone.0088589. eCollection 2014.

Abstract

Assessing the impact of global warming on the food web of the North Atlantic will require difficult-to-obtain physiological data on a key copepod crustacean, Calanus finmarchicus. The de novo transcriptome presented here represents a new resource for acquiring such data. It was produced from multiplexed gene libraries using RNA collected from six developmental stages: embryo, early nauplius (NI-II), late nauplius (NV-VI), early copepodite (CI-II), late copepodite (CV) and adult (CVI) female. Over 400,000,000 paired-end reads (100 base-pairs long) were sequenced on an Illumina instrument, and assembled into 206,041 contigs using Trinity software. Coverage was estimated to be at least 65%. A reference transcriptome comprising 96,090 unique components ("comps") was annotated using Blast2GO. 40% of the comps had significant blast hits. 11% of the comps were successfully annotated with gene ontology (GO) terms. Expression of many comps was found to be near zero in one or more developmental stages suggesting that 35 to 48% of the transcriptome is "silent" at any given life stage. Transcripts involved in lipid biosynthesis pathways, critical for the C. finmarchicus life cycle, were identified and their expression pattern during development was examined. Relative expression of three transcripts suggests wax ester biosynthesis in late copepodites, but triacylglyceride biosynthesis in adult females. Two of these transcripts may be involved in the preparatory phase of diapause. A key environmental challenge for C. finmarchicus is the seasonal exposure to the dinoflagellate Alexandrium fundyense with high concentrations of saxitoxins, neurotoxins that block voltage-gated sodium channels. Multiple contigs encoding putative voltage-gated sodium channels were identified. They appeared to be the result of both alternate splicing and gene duplication. This is the first report of multiple NaV1 genes in a protostome. These data provide new insights into the transcriptome and physiology of this environmentally important zooplankter.

摘要

评估全球变暖对北大西洋食物网的影响,将需要获取关键桡足类甲壳动物——飞马哲水蚤(Calanus finmarchicus)的生理数据,而这些数据很难获得。本文提出的从头转录组为获取此类数据提供了新资源。它由使用从六个发育阶段收集的RNA构建的多重基因文库产生:胚胎、无节幼体早期(NI-II)、无节幼体晚期(NV-VI)、桡足幼体早期(CI-II)、桡足幼体晚期(CV)和成年(CVI)雌性。在Illumina仪器上对超过4亿对末端配对读段(100个碱基对长)进行了测序,并使用Trinity软件组装成206,041个重叠群。估计覆盖率至少为65%。使用Blast2GO对包含96,090个独特成分(“comps”)的参考转录组进行了注释。40%的comps有显著的Blast比对结果。11%的comps成功地用基因本体(GO)术语进行了注释。发现许多comps在一个或多个发育阶段的表达接近零,这表明在任何给定的生命阶段,转录组的35%至48%是“沉默的”。鉴定了参与脂质生物合成途径的转录本,这些途径对飞马哲水蚤的生命周期至关重要,并研究了它们在发育过程中的表达模式。三个转录本的相对表达表明,桡足幼体晚期存在蜡酯生物合成,但成年雌性存在三酰甘油生物合成。其中两个转录本可能参与滞育的准备阶段。对飞马哲水蚤来说,一个关键的环境挑战是季节性接触含有高浓度石房蛤毒素的海洋甲藻——芬迪湾亚历山大藻,石房蛤毒素是一种阻断电压门控钠通道的神经毒素。鉴定出多个编码假定电压门控钠通道的重叠群。它们似乎是可变剪接和基因复制的结果。这是在原口动物中首次报道多个NaV1基因。这些数据为这种对环境具有重要意义的浮游动物的转录组和生理学提供了新的见解。

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