基于鳃的microRNA转录组研究中国蛤蜊对重金属镉的响应  

Identification and differential expression of gill microRNA in the Chinese surf clam(Mactra chinensis)with Cd^(2+) exposure

作　　者：张晶晶[1,2] 李宏俊[1] 秦艳杰[2] 刘敏[1,2] 叶晟[1,2] Zhang Jingjing Li Hongjun Qin Yanjie Liu Min Ye Sheng(Marine Ecology Department, National Marine Environmental Monitoring Center, Dalian 116023, China College of Fisheries and Life Science, Dalian Ocean University, Dalian 116023, China)

机构地区：[1]国家海洋环境监测中心海洋生态室,辽宁大连116023 [2]大连海洋大学水产与生命学院,辽宁大连116023

出　　处：《海洋学报》2016年第12期118-131,共14页

基　　金：国家自然科学基金(31572595);国家海洋局近岸海域生态环境重点实验室开放基金(201511)

摘　　要：中国蛤蜊(Mactra chinensis)是一种重要的经济贝类,分布于我国的辽宁和山东。近年来,生境恶化和过度捕捞导致我国蛤蜊资源量减少。我们利用Illumina Hiseq-2500平台对中国蛤蜊鳃的microRNA转录组进行测序,运用差异表达寻找对镉刺激有响应的功能microRNA。结果显示对照组获得了14 415 256条clean reads,实验组获得了15 570 111条clean reads。在这些reads中一共存在14 584 077小RNA,包括1 898 035条unique小RNA,其中对照组和实验组共有187 859条unique小RNA。两个组的小RNA文库的片段长度分布是相似的,大部分小RNA长度分布在26~27nt。在对照组文库中最丰富的片段长度是27nt,其次是28nt、26nt和23nt。在实验组文库中,数量最丰富的片段长度是26nt,其次是27nt、28nt和23nt。经过对序列前体以及结构特征的分析,发现这两个组中一共有50个microRNA,其中已知的是38个,新发现的是12个。对照组和实验组microRNA含量最丰富的片段长度是23nt。通过差异表达分析,5个microRNA基因具有显著性差异且从对照组到实验组的表达量是上调的,其余45个没有显著性的差异。把这50个序列与中国蛤蜊转录组进行比对,一共找到了542个靶基因。5个具有表达差异的基因一共比对到11个靶基因。把这11个靶基因与NCBI数据库比对,4个靶基因在COG中得到注释,1个靶基因在GO中得到注释,6个靶基因在KEGG中得到注释,11个靶基因在nr数据库中得到注释,注释到的基因有泛素蛋白连接酶E3,Wnt信号通路,G蛋白信号调控等。The Chinese surf clam （Mactra chinensis ）is an economically important clam,distributed in Lia-oning Province and Shandong Province.In recently years,because of coastal environmental deterioration and overfishing,the natural population of M.chinensis have considerably declined.In this paper,we stud-ied the microRNA transcriptome of gills,and control and experimental group were sequenced through Illu-mina Hiseq 2500 CE.The differential expression andlysis was used to find the functional microRNA re-sponse to the Cd2 ＋ exposure.Through Illumina Hi-seq 2500 CE,a total of 14 415 256 clean reads and 15 570 111 clean reads were yielded in the gill of control and experimental group respectively.A total of 14 584 077 small RNA,including 187 859 unique small RNA were yielded.The distribution of the small RNA length in the two library was similar,most of them were 26 -27 nt.27 nt was the most abundant length in control group,followded by 28 nt,26 nt,and 23 nt;26 nt was the most abundant length in ex-perimental group,and followed by 27 nt,28 nt and 23 nt.50 microRNA was found in unique small RNA, including 38 conserved and 12 novel genes.The most abundant length of microRNA in the two library was the same,23 nt.Through the analyze of differential expression analysis,the expression of 5 microRNA was induced with significantly difference,other 45 microRNA was regulated up or down without signifi-cantly difference.542 target genes were yielded when the 50 microRNA were hit to mRNA genome.And the target genes of differential expression microRNA were annotated by hitting to the NCBI database,and 4 genes hit to the COG,1 genes hit to the GO,5 genes hit to the KEGG and 11 genes hit to the nr data-base.The genes hit to the NCBI database included E3 ubiquitin-protein ligase,Wnt signaling pathway and Regulator of G-protein signaling 22.

关 键 词：中国蛤蜊 镉 MICRORNA 差异表达 功能基因 

分 类 号：S917.4[农业科学—水产科学]