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  • 周慧,吴海霞,谢伟,王芸芸,蒋琦辰,李鹏.温度和光照对克氏原螯虾生物钟基因PcClkPcCry1表达的影响[J].广西科学,2022,29(2):393-404.    [点击复制]
  • ZHOU Hui,WU Haixia,XIE Wei,WANG Yunyun,JIANG Qichen,LI Peng.Effects of Temperature and Light on the Expression of Circadian Clock Genes PcClk and PcCry1 in Procambarus clarkii[J].Guangxi Sciences,2022,29(2):393-404.   [点击复制]
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温度和光照对克氏原螯虾生物钟基因PcClkPcCry1表达的影响
周慧1,2, 吴海霞2, 谢伟2, 王芸芸2, 蒋琦辰3, 李鹏2
0
(1.南京师范大学海洋科学与工程学院, 江苏南京 210023;2.南京师范大学生命科学学院, 江苏南京 210023;3.江苏省淡水水产研究所, 江苏南京 210017)
摘要:
生物钟基因clock(Clk)和cryptochrome(Cry)参与生物内在节律调控。克氏原螯虾(Procambarus clarbii)是我国重要的水产养殖经济虾类,为探究温度、光照对克氏原螯虾生长及其生物钟基因PcClkPcCry1节律振荡表达的影响,采用实时定量PCR技术探讨克氏原螯虾PcClkPcCry1基因在脑、眼柄和肝胰腺组织中的mRNA表达模式,并探讨PcClkPcCry1基因在温度和光照周期适应过程中是否发生适应性进化。荧光定量PCR检测结果显示,克氏原螯虾的PcClk基因和PcCry1基因在脑、眼柄和肝胰腺中均有表达。在不同光照周期培养下,PcClk基因和PcCry1基因在脑、眼柄中的mRNA表达有一定的振荡节律,在25℃、12 LD的培养模式下,脑中PcClk基因和PcCry1基因mRNA表达的节律性会更明显。选择压力分析显示,经PAML和Datamonkey共同筛选出的PcCLK正选择位点为71与213,PcCRY1正选择位点为63。以上结果表明,克氏原螯虾生物钟基因PcClkPcCry1受到了选择压力,克氏原螯虾PcClk基因和PcCry1基因是内源性的节律基因。
关键词:  昼夜节律  克氏原螯虾  生物钟基因  mRNA表达  选择压力
DOI:10.13656/j.cnki.gxkx.20220526.021
投稿时间:2021-09-06
基金项目:国家虾蟹产业技术体系克氏原螯虾种质资源与遗传育种岗位项目(CARS-48),江苏省农业重大新品种创制项目(PZCZ201746),江苏省高等学校自然科学研究重大项目(19KJA330001),江苏省现代农业(克氏原螯虾) 产业技术体系项目(JATS[2020]365),江苏省研究生科研与实践创新计划项目(KYCX20_1241),江苏省水生生物资源重大专项(ZYHB16-3)和江苏省海洋渔业资源环保项目资助。
Effects of Temperature and Light on the Expression of Circadian Clock Genes PcClk and PcCry1 in Procambarus clarkii
ZHOU Hui1,2, WU Haixia2, XIE Wei2, WANG Yunyun2, JIANG Qichen3, LI Peng2
(1.College of Ocean Science and Engineering, Nanjing Normal University, Nanjing, Jiangsu, 210023, China;2.College of Life Sciences, Nanjing Normal University, Nanjing, Jiangsu, 210023, China;3.Jiangsu Freshwater Fisheries Research Institute, Nanjing, Jiangsu, 210017, China)
Abstract:
Clock (Clk) genes and cryptochrome (Cry) genes are involved in the regulation of biological intrinsic rhythm.Procambarus clarkii is an important economic aquaculture shrimp in China.In order to explore the effects of temperature and light on the growth of Procambarus clarkii and the rhythmic oscillation expression of clock genes PcClk and PcCry1,real-time quantitative PCR was used to explore the mRNA expression patterns of PcClk and PcCry1 genes in brain,eyestalk stem and hepatopancreas tissues of Procambarus clarkii. The adaptive evolution of PcClk and PcCry1 genes in the adaptation process of temperature and photoperiod was also discussed.The results of fluorescence quantitative PCR showed that PcClk and PcCry1 genes of Procambarus clarkii were both expressed in the brain,eyestalk and hepatopancreas.Under different photoperiod,the mRNA expression of PcClk and PcCry1 genes in the brain and eyestalk showed a certain oscillating rhythm,and the mRNA expression of PcClk and PcCry1 genes in the brain was more obvious under the culture mode of 25℃ and 12 LD.The selection pressure analysis showed that the positive selection sites of PcCLK screened by PAML and Datamonkey were 71 and 213,and the positive selection sites of PcCRY1 were 63.The above results indicated that the PcClk and PcCry1 genes were subjected to the selection pressure,and PcClk and PcCry1 genes were endogenous rhythm genes.
Key words:  circadian rhythm  Procambarus clarkii  circadian clock gene  mRNA expression  selection pressure

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