关键词：
白鲫;杂交子代;转录组;信号通路;SSR位点
摘 要：
[目的]研究3种白鲫杂交子代转录组学特征,为揭示鲫鲤杂交优势的分子机理提供理论依据,同时为在生产上培育出生长速度快、肉质好、适应能力强的杂交品种提供技术参考.[方法]以白鲫(♀)×黑龙江野鲤(♂)杂交子代(简称HB)、白鲫(♀)×散鳞镜鲤(♂)杂交子代(简称SB)和白鲫(♀)×兴国红鲤(♂)杂交子代(简称XB)为研究对象,利用RNA-seq高通量测序技术构建3种白鲫杂交子代转录组文库,以HiSeq PE150进行测序分析,原始序列经Trinity组装后进行功能注释(E-value<1e-5);以DESeq2 R鉴定差异表达基因,利用GOseq R和KOBAS分别对差异表达的基因进行GO和KEGG富集分析;并采用MicroSAtellite对转录本中的SSR位点进行挖掘.[结果]共组装得225858条unigenes,平均长度为668 bp,N50为938 bp,有171461条unigenes可注释到蛋白质数据库(Nr)、非冗余核苷酸数据库(Nt)、蛋白质序列数据库(SwissPort)、基因本体论(GO)、直系同源基因簇(COG/KOG)和京都基因与基因组百科全书(KEGG)数据库中,注释比例为75.92%.其中,52630条unigenes注释到NR数据库,43659条unigenes注释到SwissPort数据库,35756条unigenes注释到COG/KOG数据库,包括生化代谢、信号转导机制、防御系统和细胞结构等.差异表达基因KEGG分析结果显示,较多的差异表达基因注释到内吞作用、Jak-STAT信号通路、溶酶体、吞噬体和Wnt信号通路等免疫相关及与生长发育相关的MAPK信号通路、Hippo信号通路和背腹轴形成等通路中.此外,从获得的转录组序列中共鉴定出20272个SSR位点,大多数为二核苷酸重复基元(占62.15%).[结论]不同白鲫杂交子代间存在较多的差异表达基因,从中获得参与抗氧化、免疫和生长发育相关的通路和基因序列,且挖掘出20272个SSR位点,有助于选择性育种、分子标记开发及开展遗传多样性、遗传图谱构建和QTL定位等研究.
译 名：
Transcriptomic analysis of three Carassius auratus cuvieri hybrids
作 者：
ZHOU Da-yan;ZHANG Zhi-xin;HUANG Cai-lin;ZHAO Zhi-jie;MO Fei-long;Aquatic Species Introduction and Breeding Center of Guangxi;
关键词：
Carassius auratus cuvieri;;hybrid;;transcriptome;;signal pathway;;SSR locus
摘 要：
【Objective】Transcriptomics characteristics of three Carassius auratus cuvieri hybrids were studied to provide reference for molecular mechanism of hybrid heterosis of the hybrids,and also to offer technical support for breeding hybrids with rapid growth,quality meat and strong adaptability.【Method】Three white crucian carp hybrids i.e. C. auratus cuvier(i♀)×Cyprinus carpio haermatopterus(♂)(abbreviated as HB),C. auratus cuvier(i♀)×C. carpio L(.♂)(abbreviated as SB)and C. auratus cuvier(i♀)×C. carpio var. singuonensis(♂)(abbreviated as XB)were used as research objectives.RNA-seq high-throughput sequencing was used to construct C. auratus cuvieri hybrids transcriptome library. HiSeq PE150 was used to conduct sequencing analysis,and the function annotation on original sequence after Trinity assembling was carried out(E-value<1 e-5). Differentially expressed genes was identified by DESeq2 R,then GO and KEGG enrichment analysis of differentially expressed genes were conducted by GOseq R and KOBAS respectively. MicroSAtellite was used to dig SSR loci in transcript.【Result】A total of 225858 unigenes,with average length of 668 bp and N50 of 938 bp were generated. Of these,a total of 171461 unigenes could be annotated in protein sequence database(Nr),non-redundant nucleotide database(Nt),protein sequence database(SwissProt),Gene Ontology database(GO),Clusters of Orthologous Groups(COG/KOG)and Kyoto Encyclopedia of Genes and Genomes(KEGG)database. The annotation ratio was75.92%. There were 52630 unigenes annotated in NR database,43659 unigenes annotated in SwissPort database,35756 unigenes annotated in COG/KOG database,including biochemical metabolism,signal transduction mechanism,defensive system and cell structure. KEGG analysis of differentially expressed genes showed that more differentially expressed genes were involved in immune-related pathways such as endocytosis,Jak-STAT signaling pathway,lysosome,phagosome and Wnt signaling pathways,and growth and development related pathways such as MAPK signaling pathway,Hip-po signaling pathway and dorso-ventral axis formation. In addition,20272 SSR loci were identified from transcriptome sequence and majority of which were dinucleotide repeats units(accounted for 62.15%).【Conclusion】There are many differentially expressed genes in different hybrids. Some pathways and gene sequences involved in antioxidation,immunity,growth and development are preliminarily obtained. A total of 20272 SSR loci are dug. This result is helpful for selective breeding,molecular markers development,researches in genetic diversity,genetic map construction and QTL mapping.
