缢蛏HIF-1和PHD2基因鉴定及其在低氧胁迫下的表达OA
Identification and expression analysis of HIF-1 and PHD2 genes in razor clam(Sinonovacula constricta)under hypoxia stress
[目的]了解缢蛏在低氧环境下HIF信号通路在代谢和能量平衡调节等方面的作用.[方法]利用PCR技术克隆了缢蛏 HIF信号通路中的 4 个关键基因HIF-1α、HIF-1β、PHD2A和PHD2B,对其编码蛋白的理化性质和结构域进行预测,并对其系统发育关系进行分析.[结果]HIF-1 包含典型的 HLH、PAS、PAC、C-TAD结构域,HIF-1α特有ODDD和N-TAD结构域,ScPHD2 含有zf-MYND和P4Hc结构域;不同于其他无脊椎动物,贝类包括缢蛏PHD2 具有 2 个拷贝.利用实时荧光定量PCR(RT-qPCR)技术对HIF-1 和PHD2 在不同发育阶段、成体不同组织和低氧(0.5 和 2.0 mg/L)及干露(21℃和 4℃干露)胁迫下的表达水平进行分析.结果显示,缢蛏HIF-1 和PHD2 在卵子时期便开始表达,在检测的 6 个组织中ScHIF-1α的表达水平高于其他基因,且在鳃中表达量最高,肝胰腺次之,而ScPHD2A和ScPHD2B在6 个组织的表达量均较低.低氧胁迫下,ScHIF-1α、ScPHD2 表达量显著上调,且均在 0.5 mg/L胁迫 24 h达到最大值,而ScHIF-1β表达量变化不显著.干露胁迫下,ScHIF-1β表达量变化不显著,ScHIF-1α、ScPHD2 表达量显著上调,并同时在 4℃干露 36 h表达量达到最大值.[结论]ScHIF-1 和ScPHD2 分别具备典型的HIF和PHD家族特征,且在低氧条件下表达量升高,表明其可能参与了缢蛏低氧胁迫后应答过程.研究结果为深入开展缢蛏低氧信号通路和低氧适应机制研究提供了数据基础和参考价值.
The HIF(hypoxia-inducible factor)signaling pathway is an oxygen-sensing signaling pathway in animals and plays important roles in metabolism and energy homeostasis under hypoxic conditions.In this study,four key genes of HIF signaling pathway—HIF-1α,HIF-1β,PHD2A and PHD2B were cloned from Sinonovacula constricta using PCR.The physicochemical properties and domains of their encoded proteins were predicted,and their phylogenetic relationships were analyzed.The results showed that HIF-1 contained typical HLH,PAS,PAC,and C-TAD domains,HIF-1α specific ODDD and N-TAD domains,and ScPHD2 contained zf-MYND and P4Hc domains;unlike other invertebrates,shellfish including S.con-stricta PHD2 had two copies.Using real-time quantitative PCR(RT-qPCR),the expression levels of HIF-1 and PHD2 were further analyzed at different developmental stages,in different tissues of adults and under hypoxia stress(0.5 mg/L and 2.0 mg/L)as well as air exposure stress(21℃and 4℃)stresses.The results indicated that S.constricta HIF-1 and PHD2 were expressed from the egg stage,ScHIF-1α was expressed at higher levels than the other genes in the six tissues examined,with the highest expression in gill,followed by hepatopancreas,whereas ScPHD2A and ScPHD2B were expressed at very low levels in all six tissues.Under hypoxia stress,the expressions of ScHIF-1α and ScPHD2 were significantly up-regulated,with a greater up-regulation in the 0.5 mg/L group than in the 2 mg/L group,while ScHIF-1β expressions did not significantly changed.Under air exposure stress,ScHIF-1β expression remained unchanged,whereas ScHIF-1α and ScPHD2 expression were significantly up-regulated,with a stronger up-regulation in the 4℃air exposure group than in the 21℃group.These findings demonstrate that ScHIF-1 and ScPHD2 possessed typical characteristics of the HIF and PHD family,respectively,and their expression is induced to be up-regulated under hypoxia,suggesting that they may be involved in the response process of S.constricta after low oxygen stress.This study provide a reference for further research on the hypoxia signaling pathway and the adaptation mechanisms of S.constricta to hypoxic conditions.
李治平;姚璐;吕丽媛;董迎辉;任建峰
上海海洋大学,农业农村部淡水水产种质资源重点实验室,上海 201306||上海海洋大学,水产种质资源发掘与利用教育部重点实验室,上海 201306上海海洋大学,农业农村部淡水水产种质资源重点实验室,上海 201306||上海海洋大学,水产种质资源发掘与利用教育部重点实验室,上海 201306浙江万里学院生物与环境学院,浙江省水产种质资源高效利用技术研究重点实验室,浙江 宁波 315000浙江万里学院生物与环境学院,浙江省水产种质资源高效利用技术研究重点实验室,浙江 宁波 315000上海海洋大学,农业农村部淡水水产种质资源重点实验室,上海 201306||上海海洋大学,水产种质资源发掘与利用教育部重点实验室,上海 201306
农业科技
缢蛏HIF-1PHD2低氧干露基因表达
Sinonovacula constrictaHIF-1PHD2hypoxiaair exposuregene expression
《水产学报》 2026 (2)
65-76,12
浙江省重大科技专项(2021C02069-7)宁波市"科技创新2025"重大专项(2019B10005) Zhejiang Province Science and Technology Major Project(2021C02069-7)Ningbo City"Science and Technology Innovation 2025"Major Project(2019B10005)
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