Aja-miR-22和Aja-miR-27及其靶基因在刺参盐度胁迫后的表达模式  

Expression profiles of Aja-miR-22 and Aja-miR-27, and their target genes, in sea cucumber under salinity stress

作　　者：郭然 吴玮杰 商艳鹏 仇雪梅[1] 丁君[1] 李翔 田燚[1] GUO Ran;WU Weijie;SHANG Yanpeng;QIU Xuemei;DING Jun;LI Xiang;TIAN Yi(Key Laboratory of Mariculture&Stock Enhancement in North China’s Sea,Ministry of Agriculture and Rural Affairs,Dalian Ocean University,Dalian 116223,China)

机构地区：[1]大连海洋大学,农业农村部北方海水增养殖重点实验室,辽宁大连116223

出　　处：《中国水产科学》2020年第4期375-382,共8页Journal of Fishery Sciences of China

基　　金：国家重点研发计划项目(2018YFD0901601).

摘　　要：盐度是影响刺参(Apostichopus japonicas)生长发育最重要的环境因子之一,而microRNAs(miRNAs)可通过与靶基因的mRNA特异性结合,实现对其靶基因的表达调控。为探究Aja-miR-22和Aja-miR-27及对应的两个靶基因在刺参低盐海水胁迫下的表达情况,本实验将刺参放入盐度18低盐海水中进行胁迫,取盐度胁迫后的3 h、6 h、12 h、24 h、48 h和72 h不同时间点和正常盐度海水中刺参的体腔液,提取RNA和miRNA,利用qRT-PCR技术进行表达分析。结果表明Aja-miR-22和Aja-miR-27这两个miRNA在盐度胁迫后各时段均呈现出上调表达趋势,且都在胁迫后3 h达到最大值,分别为对照组的36倍和16倍。Aja-miR-22和Aja-miR-27在胁迫后各个表达量都均高于对照组,其中Aja-miR-22在胁迫后3 h、48 h与对照组差异极显著(P<0.01),Aja-miR-27在胁迫后3 h、48 h、72 h与对照组差异极显著(P<0.01)。靶基因棘皮动物微管相关蛋白(77 kD echinoderm microtubule-associated protein)与一般转录因子IIE亚基2(general transcription factor IIE subunit 2)的表达趋势基本一致,在胁迫后3 h降到最低值,其中棘皮动物微管相关蛋白(EMAP)在胁迫后各个时间点表达量均低于对照组,除胁迫后48 h外其余时间点均与对照组差异极显著(P<0.01),而一般转录因子IIE亚基2(general transcription factor IIE subunit 2)在胁迫后48 h和胁迫后72 h的表达量与对照组差异极显著(P<0.01)。Aja-miR-22和Aja-miR-27的表达趋势与其靶基因表达模式在24 h前均呈现负相关关系。上述研究结果说明Aja-miR-22和Aja-miR-27及其对应的靶基因在刺参的盐度适应调节机制中发挥作用。Salinity is one of the most important environmental factors affecting the growth and development of sea cucumbers.microRNAs(miRNAs)can regulate the expression of their target genes by specifically binding to target mRNA.The expression profiles of Aja-miR-22 and Aja-miR-27,and two target genes,were detected in sea cucumbers that were under low salt stress.RNA and miRNA were extracted from coelomocytes of sea cucumbers in normal salini-ty seawater and were,after 3 h,6 h,12 h,24 h,48 h,and 72 h exposure to salinity stress,analyzed by qRT-PCR.The results showed that the expression profile of Aja-miR-22 was consistent with that of Aja-miR-27 at each time point after initial salinity stress.Two miRNAs were induced to up-regulate and reached maximum expression at 3 h after stress.The expression maximums of Aja-miR-22 and Aja-miR-27 were 36 times and 16 times more than in the control group,respectively.The expression levels of Aja-miR-22 and Aja-miR-27 were higher than those of the control group under salinity stress,and Aja-miR-22 was significantly different from the control group at 3 h and 48 h after initial stress exposure(P<0.01).Aja-miR-27 was significantly different from the control group at 3 h,48 h,and 72 h under salinity stress.The expression of the target gene echinoderm microtubule-associated protein(EMAP;77 kD)was also consistent with the general transcription factor IIE subunit 2,which reached the minimum value at 3 h under salinity stress.The expression level of EMAP was lower than that of the control group at each time point after salinity stress.The general transcription factor IIE subunit 2(General Transcription Factor IIE Subunit 2)was significantly different from the control group at 48 h and 72 h under salinity stress(P<0.01).The expression trends of Aja-miR-22 and Aja-miR-27 were negatively correlated with their target gene expression patterns before 24 h.The above results indi-cated that Aja-miR-22 and Aja-miR-27,and their corresponding target genes,are involved in the salinity adaptation mechanism

关 键 词：刺参 盐度胁迫 miRNA基因表达 靶基因表达 

分 类 号：S94[农业科学—水产养殖] S917[农业科学—水产科学]