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陈洁, 董梦, 崔悦, 房翠莲, 陈旭堂, 刘子明. 低温胁迫下两种不同形态群体拟穴青蟹转录组分析[J]. 水生生物学报, 2022, 46(8): 1256-1264. DOI: 10.7541/2022.2021.129
引用本文: 陈洁, 董梦, 崔悦, 房翠莲, 陈旭堂, 刘子明. 低温胁迫下两种不同形态群体拟穴青蟹转录组分析[J]. 水生生物学报, 2022, 46(8): 1256-1264. DOI: 10.7541/2022.2021.129
CHEN Jie, DONG Meng, CUI Yue, FANG Cui-Lian, CHEN Xu-Tang, LIU Zi-Ming. TRANSCRIPTOME ANALYSIS OF TWO DIFFERENT MORPHOLOGICAL POPULATIONS OF SCYLLA PARAMAMOSAIN UNDER COLD STRESS[J]. ACTA HYDROBIOLOGICA SINICA, 2022, 46(8): 1256-1264. DOI: 10.7541/2022.2021.129
Citation: CHEN Jie, DONG Meng, CUI Yue, FANG Cui-Lian, CHEN Xu-Tang, LIU Zi-Ming. TRANSCRIPTOME ANALYSIS OF TWO DIFFERENT MORPHOLOGICAL POPULATIONS OF SCYLLA PARAMAMOSAIN UNDER COLD STRESS[J]. ACTA HYDROBIOLOGICA SINICA, 2022, 46(8): 1256-1264. DOI: 10.7541/2022.2021.129

低温胁迫下两种不同形态群体拟穴青蟹转录组分析

TRANSCRIPTOME ANALYSIS OF TWO DIFFERENT MORPHOLOGICAL POPULATIONS OF SCYLLA PARAMAMOSAIN UNDER COLD STRESS

  • 摘要: 为研究拟穴青蟹(Scylla paramamosain)不同形态群体对温度变化的适应差异, 以Sp1群体(腕节外缘中部都不具有发达的刺)和Sp2群体(至少一只螯足腕节外缘中部具有发达的刺)为研究对象, 选用低温胁迫组(8℃)和对照组(20℃)拟穴青蟹肝胰腺为实验材料, 进行转录组测序。结果显示, 测序共获得81853个Unigene, 平均长度为420 bp, N50为1460 bp, 其中22.33%为已知基因。差异表达基因筛选发现Sp1群体在低温胁迫下共有15773个Unigene显著差异表达, 其中15628个Unigene表达下调, 145个Unigene表达上调, 这些Unigene显著富集于细胞质基质等生物学过程和RNA结合、核糖体结构组分等分子功能, 并进一步富集在核糖体、剪接体和内质网蛋白质加工等信号通路。Sp2群体在低温胁迫下共有323个Unigene显著差异表达, 其中上调114个, 下调209个。这些Unigene显著富集于DNA整合等生物学过程和甾体羟化酶活性、碳酸氢盐跨膜转运体活性等分子功能, 并进一步富集在赖氨酸降解、甘氨酸、丝氨酸和苏氨酸的代谢、精氨酸和脯氨酸的代谢等代谢通路。以上结果表明, 水温从20℃骤降至8℃, Sp1群体大量基因表达下调, 生理活动大幅度降低, 这可能与寒冷的冬季青蟹停止摄食、穴居不动等行为相对应; 而Sp2群体仅调整了物质代谢, 表明Sp2群体可能对温度骤降的响应较为迟缓。利用RT-qPCR验证了转录组数据, 结果证明基于转录组分析比较两种形态群体拟穴青蟹对低温胁迫的响应是可行的, 研究结果将应用于拟穴青蟹耐寒优质品种的选育。

     

    Abstract: Scylla paramamosain is an economically important crab species mainly distribute in the southern reaches of the Yangtze River. Based on many years of breeding experience, it is speculated that different morphological populations of Scylla paramamosain have acquired different adaptations to temperature changes. In this study, Sp1 populations, with “one spine on the outer margin of the carpus of cheliped ” and Sp2 populations, with “two spines” were designated research objects, and 8℃ low temperature stress and 20℃ control groups were established. The hepatopancreas was used as a transcriptome source, and an Illumina Hiseq4000 platform was used for sequencing. A total of 81853 Unigenes were obtained by sequencing, with an average length of 420 bp, and an N50 of 1460 bp. Among these, 22.33% were known genes. Screening for differentially expressed genes identified 15773 Unigenes displaying significant differential expression in the Sp1 population under low temperature stress. Of these, 15628 Unigenes were down-regulated, and 145 Unigenes were up-regulated. Functional enrichment analysis revealed that the differentially expressed genes encode abundant cytosolic, RNA binding, and ribosome structural constituent proteins, and were further enriched in signal pathway components including ribosome, spliceosome, and protein processing components in the endoplasmic reticulum. 323 Unigenes were significantly differentially expressed in the Sp2 population under low temperature stress, of which, 209 Unigenes were down-regulated, and 114 Unigenes were up-regulated. Functional enrichment analysis revealed that the differentially expressed genes include abundant contributors to steroid hydroxylase activity, DNA integration, and bicarbonate transmembrane transporter activity, and were also enriched in signal pathways including lysine degradation, glycine, serine, and threonine metabolism, and arginine and proline metabolism. The above results indicated that as water temperature droped from 20℃ to 8℃, expression of a large number of genes was down-regulated in the Sp1 population, and physiological activities were drastically reduced. This may correspond to winter behaviors of Scylla paramamosain, including fasting and dwelling in burrows. By contrast, the Sp2 population mainly adjusted its material metabolism, indicating that the Sp2 population was relatively slow to respond to sudden temperature drops. Transcriptome data has been verified by RT-qPCR, supporting the feasibility of comparing responses of two morphological populations to low temperature stress based on transcriptome analysis. This study will inform selection of cold-tolerant and high-quality varieties for breeding of Scylla paramamosain.

     

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