An Improved RNA Extraction Method for Octocorals and Its Application in Transcriptome Analysis of Dark-Induced Bleaching Octocoral

IF 2.6 3区 生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Marine Biotechnology Pub Date : 2024-11-26 DOI:10.1007/s10126-024-10391-z
Jing-Wen Michelle Wong, An-Chi Liu, Hsuan-Tung Lin, Chuya Shinzato, Sung-Yin Yang, Shan-Hua Yang
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Abstract

Octocorals, vital components of reef ecosystems, inhabit various marine environments across diverse climate zones, spanning from tropical shallows to frigid deep-sea regions. Certain octocoral species, notably Lobophytum and Sinularia, are particularly intriguing due to their production of diverse metabolites, warranting continuous investigation. Although octocorals played the roles in coral ecosystems, the studies are rare in comparison to scleractinian corals, especially in transcriptomic and genomic data. However, RNA extraction was massively interfered by the polysaccharides and secondary metabolites produced from octocoral holobiont. For this purpose, five lysis buffer systems and two extraction processes were examined for the RNA extraction efficiency in octocorals. We found CTAB/10%SDS as a new method for RNA extraction from six different octocoral genera. Furthermore, our new method is enable to extract RNA with good quality for downstream application such as quantitative PCR and RNA sequencing. Finally, comparative transcriptomic analysis between healthy octocorals and those dark-induced bleaching corals in Lobophytum hsiehi revealed extracellular matrix and immunity-related genes may play the important roles in coral-symbiodinium symbiosis. We believe that this study’s findings and the developed RNA extraction method will serve as valuable references for future research, particularly in octocorals.

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改进的八带鱼 RNA 提取方法及其在暗诱导漂白八带鱼转录组分析中的应用
八目珊瑚是珊瑚礁生态系统的重要组成部分,栖息在不同气候带的各种海洋环境中,从热带浅滩到寒冷的深海区域。某些八目珊瑚物种,特别是Lobophytum和Sinularia,由于能产生多种代谢物而特别引人关注,值得继续研究。虽然八珊瑚在珊瑚生态系统中扮演着重要角色,但与硬骨珊瑚相比,这方面的研究却很少,特别是在转录组和基因组数据方面。然而,章鱼全缘体产生的多糖和次生代谢物会对 RNA 的提取造成严重干扰。为此,我们研究了五种裂解缓冲液体系和两种提取工艺,以检测章鱼的 RNA 提取效率。我们发现 CTAB/10%SDS 是一种从六个不同章鱼属中提取 RNA 的新方法。此外,我们的新方法还能为下游应用(如定量 PCR 和 RNA 测序)提取高质量的 RNA。最后,通过对健康章鱼和因黑暗导致白化的Lobophytum hsiehi珊瑚进行转录组学比较分析,发现细胞外基质和免疫相关基因可能在珊瑚-共生藻共生过程中发挥重要作用。我们相信,本研究的发现和所开发的 RNA 提取方法将为今后的研究,尤其是八珊瑚的研究提供有价值的参考。
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来源期刊
Marine Biotechnology
Marine Biotechnology 工程技术-海洋与淡水生物学
CiteScore
4.80
自引率
3.30%
发文量
95
审稿时长
2 months
期刊介绍: Marine Biotechnology welcomes high-quality research papers presenting novel data on the biotechnology of aquatic organisms. The journal publishes high quality papers in the areas of molecular biology, genomics, proteomics, cell biology, and biochemistry, and particularly encourages submissions of papers related to genome biology such as linkage mapping, large-scale gene discoveries, QTL analysis, physical mapping, and comparative and functional genome analysis. Papers on technological development and marine natural products should demonstrate innovation and novel applications.
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