窄螯螯虾响应高温胁迫的转录组分析
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1.新疆农业大学动物医学学院;2.海水养殖生物育种与可持续产出全国重点实验室 中国水产科学研究院黄海水产研究所 山东 青岛;3.海水养殖生物育种与可持续产出全国重点实验室 中国水产科学研究院黄海水产研究所

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Q89

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2024年科技成果转化示范专项--多营养层次综合养殖(IMTA)技术转化及应用(2024NC029)和中国水产科学研究院院级基本科研业务费“海水池塘生态养殖创新团队项目(2020TD50)”共同资助


Transcriptomic analysis of narrow-clawed crayfish (Pontastacus leptodactylus) in response to high temperature stress
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1.College of Animal Medicine, Xinjiang Agricultural University;2.National Key Laboratory of Biological Breeding and Sustainable Output of Mariculture Yellow Sea Fisheries Research Institute, Chinese Academy of Fisheries Sciences, Qingdao, Shandong, China;3.National Key Laboratory of Biological Breeding and Sustainable Output of Mariculture Yellow Sea Fisheries Research Institute, Chinese Academy of Fisheries Sciences

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    摘要:

    窄螯螯虾是一种冷淡水螯虾,具有巨大的经济发展潜力。温度作为影响水生生物生命活动重要的影响因子,近年来夏季高温气候频发,窄螯螯虾规模化养殖过程中可能会遭到高温胁迫。因此,本研究探究窄螯螯虾鳃组织在不同温度胁迫下的作用机制,分析20℃(对照)、25℃、30℃下的基因表达变化,制作病理切片,对差异基因进行生物学分析及RT-qPCR验证。结果显示,高温对鳃组织损伤随温度升高而加剧。高通量测序得CCS序列22216453个,FLNC序列同此数,去冗余后获转录本181039个。25℃与对照比,差异基因413个(上调248,下调165);30℃与对照比,差异基因1778个(上调1056,下调722);30℃与25℃比,差异基因783个(上调402,下调381)。KEGG分析显示,差异基因显著富集于代谢与信号相关通路。MDH与LDH表达下降,提示糖代谢或受破坏;HOGA1与ALDH7A1表达上升,推测糖代谢受损时,其可能借脂质代谢获能应对高温。热休克蛋白家族基因及CPT-2上调,表明其在抵御高温中起到了重要作用。研究为解析窄螯螯虾响应高温分子机制、耐高温育种等提供数据支撑。

    Abstract:

    The scientific nomenclature of the Ertysh crayfish is "narrow crayfish" (Pontastacus leptodactylus). The species under consideration is indigenous to Europe and has a wide distribution range. A taxonomic classification of species inhabiting the lakes of Eastern Europe is presented herein. The recent discovery of P.leptodactylus in the Ertysh River Basin of Xinjiang indicates a potential expansion of the species" distribution, thereby addressing a notable gap in the region"s freshwater crayfish biodiversity. To date, the extant literature on P.leptodactylus in China is limited in scope. Water temperature is a pivotal environmental variable within aquatic ecosystems, exhibiting continuous fluctuations that profoundly impact the growth, development, reproduction, and other vital life activities of aquatic organisms. Aquatic organisms exhibit a marked increase in metabolic rate when confronted with rising water temperatures, resulting in a significant escalation in oxygen demand. In instances where the dissolved oxygen level in water bodies does not meet the demand, there is an intensification of the anaerobic metabolic activities of microorganisms. This, in turn, leads to a significant accumulation of toxic metabolic products, including ammonia nitrogen,nitrite,and hydrogen sulfide. The resulting water pollution has the potential to pose a threat to the survival of aquatic organisms. Exceeding the white-spotted pike"s temperature tolerance range has been demonstrated to elicit substantial variations in several key metrics,including daily weight gain, specific growth rate, growth efficiency, and feeding efficiency. Temperature stress can have a multitude of adverse effects on aquatic animals. Temperature is a critical factor in the growth and development of diverse aquatic organisms. Consequently, it is imperative to investigate the impact of temperature on these processes. In this study, we employed the technique of transcriptome sequencing to investigate the gene expression changes in the gill tissue of P.leptodactylus under various temperature stresses. The experiment involved the analysis of three temperature gradients: 20°C (control), 25°C, and 30°C. The gill tissue were then subjected to pathological examination, and the differential genes were subjected to biological analysis. To verify the findings, RT-qPCR was employed as a confirmatory method. The differential genes were confirmed through RT-qPCR. The results demonstrated that elevated temperatures resulted in substantial damage to the gill of narrow crayfish, with the intensity of the damage increasing with rising temperatures. A comprehensive investigation was conducted to ascertain the number of CCS sequences through high-throughput sequencing and bioinformatics analysis. The investigation revealed a total of 22,216,453 CCS sequences,with an average length of 1,758 base pairs (bp) and an N50 of 1,666 bp. Furthermore, the analysis revealed that the full-length non-chimeric read (FLNC) was 22,216,453, with an average length of 1,758 bp and an N50 of 1,666 bp. Following the process of redundancy elimination, a total of 181,039 transcripts were obtained,with an average length of 2,041 base pairs (bp) and an N50 of 2,171 bp. In the experimental group B (25°C), there was a total of 413 differential genes, of which 248 were up-regulated and 165 were down-regulated in expression,in comparison to the control group A (20°C). In the experiment, Group C (30°C) was subjected to stress in comparison with Group A (20°C). The results indicated the presence of a total of 1,778 differential genes, of which 1,056 (57.9%) were found to be up-regulated, and 722 (39.9%) were found to be down-regulated in expression.

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  • 收稿日期:2025-07-11
  • 最后修改日期:2025-08-15
  • 录用日期:2025-08-15
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