刺参3个盐度相关microRNA及预测靶基因的表达模式分析  

Analysis of expression patterns of three salinity-related microRNAs and predicted target genes in sea cucumber

作　　者：王研 高思祺 匡倩瑶 刘怡宁 田燚 WANG Yan;GAO Siqi;KUANG Qianyao;LIU Yining;TIAN Yi(College of Fisheries and Life Science, Dalian Ocean University, Dalian 116023, China)

机构地区：[1]大连海洋大学水产与生命学院,辽宁大连116023

出　　处：《大连海洋大学学报》2022年第2期212-220,共9页Journal of Dalian Ocean University

基　　金：国家重点研发计划“蓝色粮仓科技创新”专项(2019YFD0900505)。

摘　　要：为研究低盐胁迫对刺参Apostichopus japonicus(16.38 g±1.27 g)体内离子浓度及钠钾ATP酶活力的影响,以及低盐胁迫与miRNA间的调控关系,试验取低盐度(18)胁迫0(对照)、6、24、48 h时的刺参体腔液,测定其钠、氯、钾离子浓度及钠钾ATP酶活力。结果表明:低盐胁迫后,刺参体腔液中钠离子浓度在6 h时显著升高(P<0.05),随胁迫时间的延长钠离子浓度有所降低,氯离子和钾离子浓度均显著低于对照组(P<0.05)且均在6 h时浓度最低,钠钾ATP酶活力在6 h时低于对照组,在24 h时出现最低值;microRNAs(miRNAs)前体序列和成熟序列分析显示,miR-2011、miR-124和miR-2010均能形成稳定的茎环结构,序列相对保守;通过miRNAs与靶基因间的序列分析,获得3个差异表达的miRNAs(miR-2011、miR-2010和miR-124)及互作的靶基因;miR-2011、miR-2010和miR-124的miRNA表达量在盐度胁迫后均呈现上调,miR-2011和miR-2010的表达量在低盐胁迫48 h时达到最大,分别为对照组(0 h)的80倍和24倍;序列预测分析获得miR-2011的靶基因为PPM1L和PBK,miR-2010的靶基因为PPM1L,miR-124的靶基因为EGF3、IMPA1。研究表明,刺参miR-124、miR-2011和miR-20103个盐度相关的miRNAs序列相对保守,且在盐度胁迫后均能够被诱导表达,从而参与刺参盐度胁迫的响应过程。Na+,Cl-,and K+ion concentrations and sodium potassium ATPase activity were determined in the coelomic fluid of sea cucumber Apostichopus japonicus with body mass of(16.38±1.27)g exposed to a salinity of 18 in 0,6,24 and 48 h.The results indicated that the sodium ion concentration increased significantly at 6 h after salinity stress(P<0.05),and then decreased with the stages of stress time.The lower concentrations of chloride and potassium ions were presented in the sea cucumber exposed to salinity stress than those of the control group(P<0.05),with the minimum at 6 h.The activity of sodium-potassium ATPase was lower than that of the control group at 6 h,with the minimum at 24 h.That miR-2011,miR-124 and miR-2010 formed a stable stem-loop structure based on the precursor sequences and mature sequences,which indicated that the three miRNAs were relatively conservative.Three differential expression in miRNAs(miR-2011,miR-2010 and miR-124)were presented and interacting target genes were obtained through miRNAs and target genes binding sites.Quantitative expression profiles showed that the miR-2011,miR-2010 and miR-124 were up-regulated in the sea cucumber exposed to salinity stress,compared to the maximal expression levels of miR-2011 and miR-2010 presented in 48 h under low-salt stress,80 times and 24 times compared to control group(0 h),respectively.The target genes of miR-2011 were PPM1L and PBK,the miR-2010 to PPM1L,and the miR-124 to EGF3 and IMPA1.The findings indicate that the three miRNAs related to salinity genes are conservative and involved in regulation of salinity stress in sea cucumber.

关 键 词：刺参 盐度胁迫 miRNA序列分析 miRNA基因表达 

分 类 号：S917.4[农业科学—水产科学]