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硫化物胁迫对缢蛏血液${\rm {SO}} _4^{2-}$浓度及SULT1B1-12基因表达的影响

孙慧妙 沈伟良 陈彩芳 林志华 韩庆喜

孙慧妙,沈伟良,陈彩芳,等. 硫化物胁迫对缢蛏血液\scriptsize${\rm {SO}} _4^{2-} $浓度及SULT1B1-12基因表达的影响[J]. 海洋学报,2023,45(1):62–70 doi: 10.12284/hyxb2023004
引用本文: 孙慧妙,沈伟良,陈彩芳,等. 硫化物胁迫对缢蛏血液\scriptsize${\rm {SO}} _4^{2-} $浓度及SULT1B1-12基因表达的影响[J]. 海洋学报,2023,45(1):62–70 doi: 10.12284/hyxb2023004
Sun Huimiao,Shen Weiliang,Chen Caifang, et al. Effects of sulfide stress on blood \scriptsize${{\rm {SO}} _4^{2-}} $ concentration and SULT1B1-12 gene expression in Sinonovacula constricta[J]. Haiyang Xuebao,2023, 45(1):62–70 doi: 10.12284/hyxb2023004
Citation: Sun Huimiao,Shen Weiliang,Chen Caifang, et al. Effects of sulfide stress on blood \scriptsize${{\rm {SO}} _4^{2-}} $ concentration and SULT1B1-12 gene expression in Sinonovacula constricta[J]. Haiyang Xuebao,2023, 45(1):62–70 doi: 10.12284/hyxb2023004

硫化物胁迫对缢蛏血液${\rm {SO}} _4^{2-}$浓度及SULT1B1-12基因表达的影响

doi: 10.12284/hyxb2023004
基金项目: 浙江省公益技术应用研究项目(LGN22C190025);国家重点研发计划“蓝色粮仓”科技创新课题(2018YFD0901405,2020YFD0900802);财政部和农业农村部:国家现代农业产业技术体系项目(CARS-49);浙江省科技重点研发计划(2019C02054,2021C02069-7)。
详细信息
    作者简介:

    孙慧妙(1996-),女,浙江省衢州市人,研究方向为滩涂贝类环境适应机制。E-mail:2075243655@qq.com

    通讯作者:

    陈彩芳,女,博士,副教授,硕士生导师,主要从事海洋生物环境适应性研究。E-mail: chencaifang@zwu.edu.cn

  • 中图分类号: Q955;Q786

Effects of sulfide stress on blood ${{\bf {SO}} _4^{2-}} $ concentration and SULT1B1-12 gene expression in Sinonovacula constricta

  • 摘要: 作为典型的埋栖型滩涂贝类,缢蛏(Sinonovacula constricta)常暴露在富含硫化物的环境中,并表现出较强的硫化物耐受能力。胞质磺基转移酶1B1(SULT1B1)位于硫代谢途径下游,是催化磺化反应的关键酶,在甲状腺激素(THs)等内源性物质的生物转化过程中发挥重要作用。为研究ScSULT1B1-12基因在缢蛏耐硫中的作用,本研究采用生物信息学方法分析了其序列特征,并结合血液中${\rm {SO}} _4^{2-} $浓度变化,开展其组织表达及不同浓度(50 μmol/L、150 μmol/L、300 μmol/L)硫化物胁迫72 h后的表达特征研究。结果表明,ScSULT1B1-12基因全长cDNA为1 100 bp,含有 897 bp的开放阅读框,编码298个氨基酸。序列分析表明,ScSULT1B1-12含有4个催化活性位点(56K、104N、106H和134A)、1个N端的PAPS结合域(YPKSGTXW)、1个C端的PAPS结合和二聚化域(RKGXXGDWKNXFTVXXE),表明其在结构上具有催化磺化反应的能力。组织分布表明,ScSULT1B1-12基因在鳃中高表达,其次为闭壳肌和肝胰腺。硫化物胁迫后缢蛏血液中${\rm {SO}} _4^{2-} $浓度呈下降趋势,ScSULT1B1-12基因的表达模式也在波动中呈下降趋势,表明硫酸盐可进一步被活化生成磺化反应的供体,而ScSULT1B1-12介导的磺化反应受抑制后可使缢蛏体内THs保持在一定水平,以加强其代谢机能和免疫功能,使机体适应高硫化物的不良环境。
  • 图  1  硫化物胁迫下缢蛏血液$ {{\rm {SO}}_4^{2-}} $浓度的变化

    同类型字母中不同字母表示同一胁迫浓度下不同时间点间的数据差异显著(p<0.05)

    Fig.  1  Changes in $ {{\rm {SO}}_4^{2-}} $ concentration of Sinonovacula constricta under sulfide stress

    Different letters of the same type represented significant changes among different time at the same exposure concentration (p<0.05)

    图  2  ScSULT1B1-12基因全长cDNA序列及其推导的氨基酸序列

    红色加粗字体为起始密码子、终止密码子;阴影部分为蛋白的保守结构域,加粗字体和下划线标注表示PAPS结合域;黄色加粗字体和下划线标注表示PAPS结合和二聚化域;蓝色加粗字体为氨基酸活性位点;左侧数字为核苷酸和氨基酸的位置

    Fig.  2  Full-length cDNA sequence of the ScSULT1B1-12 gene and its deduced amino acid sequences

    Red bold font indicated the initiation codon and stop codon, the shaded part indicated the conserved domain of the protein, the bold font and underlined mark indicated the PAPS binding domains; the yellow bold font and underlined mark indicated PAPS binding and dimerization domains; the blue bold font showed the catalytic active sites; the numbers on the left indicate the positions of nucleotide and amino acid

    图  3  11种动物SULT1B1氨基酸序列的多重比较

    绿色箭头表示催化活性位点;红框表示PAPS结合域;蓝框表示PAPS结合和二聚化域;所使用序列的物种名及基因登录号见附表S1

    Fig.  3  Multiple alignments of the amino acid sequences of SULT1B1 in eleven animals

    The green arrows indicated the catalytic active site; the red box indicated the PAPS binding domain; the blue box indicated the PAPS binding and dimerization domain; the species name and the corresponding accession numbers of the sequences used are shown in appendix Table S1

    图  4  邻接法构建的ScSULT1B1-12系统进化树

    所使用序列的基因登录号见附表S1

    Fig.  4  The ScSULT1B1-12 phylogenetic tree constructed by neighbor-joining method

    Accession numbers of the sequences used in construction of tree are shown in appendix Table S1

    图  5  ScSULT1B1-12基因在缢蛏不同组织中的表达

    不同字母表示组织间的数据差异显著(p<0.05)

    Fig.  5  The expression of ScSULT1B1-12 gene in different tissues of Sinonovacula constricta

    Different letters represented significant changes among different tissues (p<0.05)

    图  6  硫化物胁迫下ScSULT1B1-12基因在缢蛏鳃(M)和肝胰腺(m)中的表达特征

    同类型字母中不同字母表示同一胁迫浓度下不同时间点间的数据差异显著(p<0.05)

    Fig.  6  The expression characteristics of the ScSULT1B1-12 gene in gill (M) and hepatopancreas (m) of Sinonovacula constricta under sulfide stress

    Different letters of the same type represented significant changes among different time at the same exposure concentration (p<0.05)

    表  1  实验所用引物及其序列

    Tab.  1  Primers and their sequences used in the experiment

    引物名称序列(5'-3'
    SULT1B1-12-FCAAATCCGAATGGAAAGGCGG
    SULT1B1-12-RCAACAGAATCTGTATGTGAAG
    RS9-FTGAAGTCTGGCGTGTCAAGT
    RS9-RCGTCTCAAAAGGGCATTACC
    下载: 导出CSV
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  • 表S1 物种拉丁文学名及其NCBI登录号对照表.doc
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  • 收稿日期:  2022-03-25
  • 修回日期:  2022-06-24
  • 网络出版日期:  2022-11-04
  • 刊出日期:  2023-01-09

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