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| 泥蚶低氧胁迫后血细胞转录组及耐低氧相关基因的筛选与分析 |
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张阳1,2, 金铭2, 刘宏星2, 郑赟2, 詹宇1,2, 李凌云2, 程浩翔2, 包永波2,3, 彭志兰2,3, 张晓林4
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1.上海海洋大学 水产与生命学院, 上海 201306;2.浙江万里学院 生物与环境学院, 浙江 宁波 315100;3.浙江万里学院 宁海海洋种业研究院, 浙江 宁波 315600;4.浙江海洋大学 海洋科学与技术学院, 浙江 舟山 316022
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| 摘要: |
| 泥蚶(Tegillarca granosa)是一种比较特别的具有较强耐低氧能力的贝类,但目前其耐低氧分子调控机理尚不知。为探究泥蚶耐受低氧的分子调控机理,本研究对低氧胁迫下的泥蚶(Tegillarca granosa)血细胞转录组中富集的信号通路及生物学过程进行分析,并初步筛选分析耐低氧基因。对低氧(DO=0.5 mg/L)胁迫6、24、72、120 h的泥蚶血细胞进行转录组测序(RNA-seq)并开展生物信息学分析,筛选出差异基因集,并对胁迫72、120 h的DEGs进行GO富集分析、KEGG通路分析,采用qPCR方法检测7个DEGs的表达量并与转录组比较。结果显示从6 h开始到120 h的4个时间点的DEGs数量呈增多的趋势,GO功能分析主要富集在JUN激酶活性的负调控、蛋白质水解的负调控及免疫系统进程等主动抗凋亡、抗逆进程;KEGG通路分析主要富集在胰岛素及胰腺分泌的信号通路、HIF-1通路、钙信号相关通路和细胞凋亡相关通路。qPCR检测结果显示,7个基因的表达上调/下调趋势与转录组测序一致,证实了转录组测序结果的可靠性。本研究推测胰腺分泌信号通路、钙信号通路及凋亡通路在泥蚶耐受低氧过程中发挥重要作用,与这些通路相关的JUN、PIM-3、TRIM45、MR-1B、CCKR在耐低氧过程中起到关键作用,可为以后贝类耐低氧的调控机理探究及耐低氧品系的分子选育提供参考。 |
| 关键词: 泥蚶 耐低氧 血细胞 差异表达基因 转录组测序 |
| DOI：10.11759/hykx20220708001 |
| 分类号:Q955;S917.4 |
| 基金项目:浙江省公益技术应用研究计划项目(LGN21C190012);浙江省农业(水产)新品种选育重大科技专项子课题(2021C02069-7-03);海洋科学浙江省一流学科开放课题(OFMS007) |
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| Blood cell transcriptome and screening, analysis of genes related to hypoxia tolerance of Tegillarca granosa |
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ZHANG Yang1,2, JIN Ming2, LIU Hong-xing2, ZHENG Yun2, ZHAN Yu1,2, LI Ling-yun2, CHENG Hao-xiang2, BAO Yong-bo2,3, PENG Zhi-lan2,3, ZHANG Xiao-lin4
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1.College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China;2.College of Biological and Environmental Sciences, Zhejiang Wanli University, Ningbo 315000, China;3.Ninghai Institute of Mariculture Breeding and Seed Industry, Zhejiang Wanli University, Ningbo 315600, China;4.Marine Science and Technology College, Zhejiang Ocean University, Zhoushan 316022, China
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| Abstract: |
| Tegillarca granosa is a shellfish with a high tolerance to hypoxic conditions. However, not much is known about the molecular mechanism of tolerance. The study aims to understand the molecular mechanism of T. granosa by analyzing differentially expressed genes (DEGs) and changes in signaling pathways of blood cell transcripts under hypoxic and normoxic conditions. This, in turn, helps screen the hypoxia tolerance genes. Transcriptome sequencing (RNA-seq) and bioinformatics analysis of T. granosa blood cells were performed under hypoxic conditions at 6, 24, 72, and 120 h and screened for differential gene sets. GO enrichment and KEGG pathway analyses were performed on DEGs under hypoxic conditions at 72 and 120 h. The expression levels of seven DEGs were measured using qPCR and compared with the transcriptome. The results showed an increase in DEGs at four time points from 6 to 120 h. GO analysis of DEGs was mainly concentrated in some active antiapoptotic and antistress processes, such as negative regulation of JUN kinase activity, proteolysis, and immune system processes. KEGG pathway analysis revealed that DEGs were mainly enriched in insulin and pancreatic secretion signaling pathways, HIF-1 pathway, calcium signaling pathway, and apoptosis pathway. qPCR results showed that the upregulation and downregulation of seven genes were consistent with that of RNA-seq, which, in turn, confirmed the reliability of RNA-seq results. This study speculated that the pancreatic secretion signaling pathway, calcium signaling pathway and apoptosis pathway in blood clams play a key role in tolerance to hypoxic conditions. JUN, PIM-3, TRIM45, MR-1B, and CCKR related to these pathways are significant for tolerance to hypoxia. This can further help in the exploration of the regulatory mechanism and molecular breeding of hypoxia tolerant strains of the shellfish T. granosa. |
| Key words: Tegillarca granosa hypoxia blood cell differentially expressed genes RNA-seq |
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