不同温度作用下草鱼胚胎的发育及hsp70基因表达  

Embryonic Development and Expression Level of hsp70 Gene of Grass Carp Ctenopharyngodon idella under Different Temperatures

作　　者：邱裕明 林旺 郭红会 张策 李莉 李大鹏 QIU Yuming;LIN Wang;GUO Honghui;ZHANG Ce;LI Li;LI Dapeng(National Experimental Teaching Demonstration Center of Aquaculture,Hubei Provincial Engineering Laboratory of Pond Healthy Aquaculture,College of Fisheries,Huazhong Agricultural University,Wuhan 430070,China)

机构地区：[1]华中农业大学水产学院,池塘健康养殖湖北省工程实验室,水产养殖国家级实验教学示范中心,湖北武汉430070

出　　处：《水产科学》2021年第6期870-876,共7页Fisheries Science

基　　金：国家重点研发计划项目(2019YFD0900303);财政部和农业农村部现代农业产业技术体系项目(CARS-45);中央高校基本科研业务费专项资金资助项目(2662015PY119).

摘　　要：为研究温度对草鱼胚胎发育的影响及其机制,将人工授精获得的草鱼胚胎分别在22、25、28、31℃温度下孵化,测定体长,记录孵化时长、孵化率及畸形率,计算有效积温,并运用实时荧光定量PCR技术测定孵化仔鱼体内热休克蛋白70(hsp70)基因的相对表达量。试验结果显示,22~28℃,草鱼胚胎孵化率随温度升高而升高(P<0.05),畸形率随温度升高而降低(P<0.05),但在31℃时孵化率显著降低(P<0.05),仅为(31.51±1.84)%,畸形率显著升高(P<0.05),为(90.00±5.79)%,温度(x)与孵化率(y)之间的回归方程为y=-0.0192x^(2)+0.9695x-11.246(r^(2)=0.872),温度(x)与畸形率(y)之间的回归方程为y=0.0271x^(2)-1.3608x+17.035(r^(2)=0.931);随着温度升高,草鱼胚胎孵化耗时显著减少(P<0.05),孵化的有效积温显著增加(P<0.05),温度系数Q_(10)为1.77~2.27;同时,初孵仔鱼体长随温度升高呈先升后降的趋势,在28℃组达到最大值,温度(x)与体长(y)之间的回归方程为y=-0.0357x^(2)+1.9394x-21.138(r^(2)=0.999);初孵仔鱼hsp70基因相对表达量也在28℃组达到最高。本试验结果表明,温度升高能在一定程度内促使草鱼激活体内热休克蛋白以调节稳态,促进胚胎的发育,且草鱼胚胎孵化的最适温度带为25~28℃。In order to explore the effect and mechanism of water temperature on embryonic development,the embryos of grass carp Ctenopharyngodon idella obtained by artificial insemination were incubated at different temperatures(22℃,25℃,28℃,and 31℃)in the present study.The incubation duration,hatching rate and deformity rate as well as the effective accumulative temperature were recorded and calculated along with the body length of the newly hatched larvae and the expression levels of hsp70 gene in the newly hatched larvae were also detected by using real-time quantitative PCR.The results showed that the hatching rate of embryos was increased significantly with increasing exposure temperature between 22-28℃,while the deformity rate significantly was decreased.On the contrary,the hatching rate was significantly decreased [(31.51±1.84)%](P<0.05)and the deformity rate was increased significantly [(90.00±5.79)%](P<0.05)at 31℃.The regression equation between temperature and hatching rate was described as y=-0.0192 x^(2)+0.9695 x-11.246(r^(2)=0.872),and the regression equation between temperature and deformity rate was expressed as y=0.0271 x^(2)-1.3608 x+17.035(r^(2)=0.931).Also,the hatching duration was significantly decreased in a time-dependent way and the effective accumulative temperature was significantly elevated between 22-28℃.The value of Q_(10)was changed from 1.77 to 2.27.In the newly hatched larvae,the body length was increased within the range of 22-28℃but decreased at 31℃,and the regression equation between temperature and body length was expressed as y=-0.0357 x^(2)+1.9394 x-21.138(r^(2)=0.999).The relative mRNA expression for hsp70 gene was higher at 28℃than that in other groups(P<0.05).The findings indicated that increasing temperature promoted the activation of heat shock protein in grass carp embryos to some extent,which contributed to regulate homeostasis and promote embryonic development,with the range of optimal temperature of embryonic development of 25-28℃for grass carp.

关 键 词：草鱼 温度 胚胎发育 HSP70基因 

分 类 号：S917.4[农业科学—水产科学]