小黄鱼肝脏中hsp90b1和hspb1对温度胁迫的响应  

Response of hsp90b1 and hspb1 to Temperature Stress in the Liver of Larimichthys polyactis

作　　者：储天琪 刘峰 陈红林 詹炜 王梦洁 秦高婵 楼宝 徐万土 CHU Tian-Qi;LIU Feng;CHEN Hong-Lin;ZHAN Wei;WANG Meng-Jie;QIN Gao-Chan;LOU Bao;XU Wan-Tu(Institute of Hydrobiology,Zhejiang Academy of Agricultural Sciences,Hangzhou 310021 China;School of Fishery,Zhejiang Ocean University,Zhoushan 316022,China;Xiangshan Harbor Aquatic Seed Co.,Ltd.,Ningbo 315700,China)

机构地区：[1]浙江省农业科学院水生生物研究所,杭州310021 [2]浙江海洋大学水产学院,舟山316022 [3]象山港湾水产苗种有限公司,宁波315700

出　　处：《农业生物技术学报》2022年第3期528-538,共11页Journal of Agricultural Biotechnology

基　　金：国家重点研发计划“蓝色粮仓科技创新”重点专项(2018YFD0901204);国家自然科学基金(32102765);浙江省重点研发计划(2020C02015,2021C02055);象山县科技计划(2019C0001)。

摘　　要：热休克蛋白(heat shock proteins,HSPs)与鱼类响应温度胁迫密切相关。为研究HSPs家族基因(hsp90b1和hspb1)在重要海水鱼类小黄鱼(Larimichthys polyactis)温度胁迫过程中的响应特点,本研究对hsp90b1、hspb1进行克隆分析,采用实时荧光定量PCR技术对2个基因进行组织表达量测定,通过急性和慢性2种温度处理方式对2个基因进行温度响应分析。结果显示,hsp90b1(GenBank No.OK340652)的CDS序列长度为2406 bp,编码801个氨基酸;hspb1(GenBank No.OK340653)的CDS序列长度为609 bp,编码202个氨基酸。2个基因在小黄鱼的肝脏、脑、鳃、肠、肌肉、皮肤和肾脏等组织中均有表达,其中hsp90b1在皮肤中的表达量最高,而hspb1在肌肉中表达量最高。在自然水温为20℃(对照组)时,对小黄鱼分别进行急性高温(32℃)和低温(6℃)胁迫处理,发现急性温度处理可诱导hsp90b1显著高表达,而hspb1则表现出显著的高温高表达、低温低表达的特点(P<0.05)。以13℃(自然水温)为对照,对小黄鱼分别进行6、8、16和20℃长期温度处理(7 d),发现肝脏中hsp90b1表达水平均显著高于对照组(P<0.05),同时高温组(16和20℃)的表达水平要高于低温组(6和8℃);与hsp90b1不同,hspb1仍呈现出高温高表达、低温低表达的特点(P<0.05)。上述研究表明,hsp90b1和hspb1在小黄鱼温度胁迫过程中出现明显的差异表达,表明其在温度胁迫响应过程中发挥着重要作用。本研究结果为探究小黄鱼响应温度胁迫的分子机制提供了参考。Heat shock proteins(HSPs)are closely related to fish response to temperature stress.To investigate the response characteristics of the heat shock proteins family genes,hsp90b1 and hspb1,in Larimichthys polyactis subjected to temperature stress,In this study,full-length CDS of hsp90b1 and hspb1 in L.polyactis was cloned,and detected their spatiotemporal expression characteristics and expression patterns after acute and long-term temperature stress.The full-length CDS sequence of hsp90b1(GenBank No.OK340652)was 2406 bp,encoding a predicted 801 amino acid.While the full-length CDS sequence of hspb1(GenBank No.OK340653)was 609 bp encoding 202 amino acid.Tissue expression analysis results showed that the hsp90b1 gene expression is the highest in the skin,while hspb1 gene expression was the highest in the muscle.As the natural water temperature was 20℃(control group),L.polyactis was treated with acute high(32℃)and low(6℃)temperature stress.The results showed that the acute temperature stress could induce significant high expression of hsp90b1 in the liver.Differently,hspb1 expressed higher with high temperature and expressed lower with low temperature(P<0.05).During long-term temperature treatments(with 13℃as the control,fish was treated at 6,8,16 and 20℃,respectively and maintained for 7 d).It was found that the expression of hsp90b1 in the liver of the 4 treatment groups was significantly higher than that in the control group,which temperature was 13℃(P<0.05),while the expression level of the high-temperature group(16℃and 20℃)was higher than that in the low-temperature group(6℃and 8℃);Unlike hsp90b1,hspb1 still showed the characteristics of high expression at high temperature and low expression at low temperature(P<0.05).The present study indicated that hsp90b1 and hspb1 were significantly differentially expressed in L.polyactis during temperature stress,suggested that hsp90b1 and hspb1 played an important role in the response to temperature stress.The results of this study provide an important fo

关 键 词：小黄鱼 热休克蛋白(HSPs) 温度胁迫 基因克隆 qPCR 

分 类 号：S965.323[农业科学—水产养殖]