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RNA-seq 分析两种泥蟹形态对低温胁迫的差异响应。

RNA-seq analysis of the differential response to low-temperature stress in two morphs of mud crabs (Scylla paramamosain).

机构信息

College of Ecology, Lishui University, Lishui 323000, China.

College of Ecology, Lishui University, Lishui 323000, China.

出版信息

Comp Biochem Physiol Part D Genomics Proteomics. 2022 Sep;43:101010. doi: 10.1016/j.cbd.2022.101010. Epub 2022 Jun 22.

Abstract

The mud crab (Scylla paramamosain) is an economically important crab species that is distributed along the southern coast of China. Previously we reported that various morphological types of mud crabs differentially adapt to temperature changes. This study investigates the effects of temperature on morphs with one or two spines (Sp1 and Sp2, respectively) on the outer margin of the carpus of cheliped at a low temperature (8 °C) and a control temperature (20 °C). The hepatopancreas were the transcriptome source, and a total of 81,853 unigenes were obtained by sequencing, with an average length of 420 bp, and an N50 of 1460 bp. Of these, 22.33 % were known genes. Under low-temperature stress, there were 361 differentially expressed genes (DEGs) between the two morphs; 96 and 265 were up- and down-regulated genes, respectively. There were no DEGs between the morphs at 20 °C. Functional enrichment analysis revealed that the DEGs encoded abundant metallocarboxypeptidase activity, extracellular space, proteolysis, and sequence-specific DNA binding, and were further enriched in signal pathway components, including ubiquinone and other terpenoid-quinone biosynthesis, phenylalanine metabolism, MAPK signaling pathway, apoptosis, and other signaling pathways. The gene expressions of acid phosphatase (ACP), chymotrypsin (CHY), serine carboxypeptidase (SCP), and trypsin (TRY) under low-temperature stress were detected. The expressions of ACP, CHY, and SCP gradually decreased with time. The ACP, CHY, SCP, and TRY gene expression of the Sp1 morph was lower than that of the Sp2 morph; however, the ACP gene expression of the Sp1 morph was higher than that of the Sp2 morph at several time points. In conclusion, adaptability differences between the morphs to low-temperature stress were confirmed, which will enable the selection of cold-tolerant, high-quality varieties of mud crabs for breeding.

摘要

泥蟹(Scylla paramamosain)是一种经济上重要的蟹类,分布在中国南部沿海。先前我们报道过,泥蟹的各种形态类型对外界温度变化有不同的适应能力。本研究调查了低温(8°C)和对照温度(20°C)下,外测腕节有 1 或 2 根刺(分别为 Sp1 和 Sp2)的形态对温度的影响。肝胰腺是转录组的来源,通过测序共获得 81853 条全长非编码 RNA(unigenes),平均长度为 420bp,N50 为 1460bp。其中,22.33%是已知基因。在低温胁迫下,两种形态之间有 361 个差异表达基因(DEGs);分别有 96 个和 265 个上调和下调基因。在 20°C 时,两种形态之间没有差异表达基因。功能富集分析表明,DEGs 编码丰富的金属羧肽酶活性、细胞外空间、蛋白水解和序列特异性 DNA 结合,并进一步富集在信号通路成分中,包括泛醌和其他萜类醌生物合成、苯丙氨酸代谢、MAPK 信号通路、细胞凋亡和其他信号通路。在低温胁迫下检测到酸性磷酸酶(ACP)、糜蛋白酶(CHY)、丝氨酸羧肽酶(SCP)和胰蛋白酶(TRY)的基因表达。随着时间的推移,ACP、CHY 和 SCP 的表达逐渐下降。Sp1 形态的 ACP、CHY、SCP 和 TRY 基因表达均低于 Sp2 形态;然而,在几个时间点,Sp1 形态的 ACP 基因表达高于 Sp2 形态。总之,证实了形态对低温胁迫的适应性差异,这将有助于选择耐寒、高质量的泥蟹品种进行养殖。

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