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对以产生氧化脂质的硅藻——海洋骨条藻为食的海洋桡足类动物赫氏哲水蚤转录组的见解。

Insights into the transcriptome of the marine copepod Calanus helgolandicus feeding on the oxylipin-producing diatom Skeletonema marinoi.

作者信息

Carotenuto Ylenia, Dattolo Emanuela, Lauritano Chiara, Pisano Fabio, Sanges Remo, Miralto Antonio, Procaccini Gabriele, Ianora Adrianna

机构信息

Stazione Zoologica Anton Dohrn, Villa Comunale, Napoli, Italy.

Stazione Zoologica Anton Dohrn, Villa Comunale, Napoli, Italy.

出版信息

Harmful Algae. 2014 Jan;31:153-162. doi: 10.1016/j.hal.2013.11.002. Epub 2013 Dec 11.

Abstract

Diatoms dominate productive regions in the oceans and have traditionally been regarded as sustaining the marine food chain to top consumers and fisheries. However, many of these unicellular algae produce cytotoxic oxylipins that impair reproductive and developmental processes in their main grazers, crustacean copepods. The molecular mode of action of diatoms and diatom oxylipins on copepods is still unclear. In the present study we generated two Expressed Sequence Tags (ESTs) libraries of the copepod Calanus helgolandicus feeding on the oxylipin-producing diatom Skeletonema marinoi and the cryptophyte Rhodomonas baltica as a control, using suppression subtractive hybridization (SSH). Our aim was to investigate differences in the transcriptome between females fed toxic and non-toxic food and identify differentially expressed genes and biological processes targeted by this diatom. We produced 947 high quality ESTs from both libraries, 475 of which were functionally annotated and deposited in GenBank. Clustering and assembling of ESTs resulted in 376 unique transcripts, 200 of which were functionally annotated. Functional enirchment analysis between the two SSH libraries showed that ESTs belonging to biological processes such as response to stimuli, signal transduction, and protein folding were significantly over-expressed in the S. marinoi-fed C. helgolandicus compared to R. baltica-fed C. helgolandicus library. These findings were confirmed by RT-qPCR analysis. In summary, 2 days of feeding on S. marinoi activated a generalized Cellular Stress Response (CSR) in C. helgolandicus, by over-expressing genes of molecular chaperones and signal transduction pathways that protect the copepod from the immediate effects of the diatom diet. Our results provide insights into the response of copepods to a harmful diatom diet at the transcriptome level, supporting the hypothesis that diatom oxylipins elicit a stress response in the receiving organism. They also increase the genomic resources for this copepod species, whose importance could become ever more relevant for pelagic ecosystem functioning in European waters due to global warming.

摘要

硅藻在海洋的高产区域占主导地位,传统上被认为维系着海洋食物链直至顶级消费者和渔业。然而,这些单细胞藻类中有许多会产生细胞毒性氧化脂质,损害其主要捕食者——甲壳类桡足类动物的生殖和发育过程。硅藻及其氧化脂质对桡足类动物的分子作用模式仍不清楚。在本研究中,我们利用抑制性消减杂交(SSH)技术,构建了以产生氧化脂质的硅藻海链藻(Skeletonema marinoi)为食的哲水蚤(Calanus helgolandicus)以及作为对照的隐藻波罗红胞藻(Rhodomonas baltica)的两个表达序列标签(EST)文库。我们的目的是研究摄食有毒和无毒食物的雌性哲水蚤转录组的差异,识别差异表达基因以及该硅藻所靶向的生物学过程。我们从两个文库中获得了947个高质量的EST,其中475个进行了功能注释并提交到GenBank。EST的聚类和组装产生了376个独特的转录本,其中200个进行了功能注释。两个SSH文库之间的功能富集分析表明,与以波罗红胞藻为食的哲水蚤文库相比,在以海链藻为食的哲水蚤中,属于对刺激的反应、信号转导和蛋白质折叠等生物学过程的EST显著过度表达。这些发现通过RT-qPCR分析得到了证实。总之,以海链藻为食2天会通过过度表达分子伴侣和信号转导途径的基因,激活哲水蚤的全身性细胞应激反应(CSR),从而保护桡足类动物免受硅藻饮食的直接影响。我们的结果在转录组水平上深入了解了桡足类动物对有害硅藻饮食的反应,支持了硅藻氧化脂质在受体生物中引发应激反应的假说。它们还增加了这种桡足类物种的基因组资源,由于全球变暖,其重要性对于欧洲水域的远洋生态系统功能可能变得越来越重要。

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