title: Transcriptome analysis of orange-spotted grouper (Epinephelus coioides) spleen in response to Singapore grouper iridovirus.
abstract: BACKGROUND
  Orange-spotted grouper (Epinephelus coioides) is an economically important marine fish cultured in China and Southeast Asian countries. The emergence of infectious viral diseases, including iridovirus and betanodavirus, have severely affected food products based on this species, causing heavy economic losses. Limited available information on the genomics of E. coioides has hampered the understanding of the molecular mechanisms that underlie host-virus interactions. In this study, we used a 454 pyrosequencing method to investigate differentially-expressed genes in the spleen of the E. coioides infected with Singapore grouper iridovirus (SGIV).
  
  
  RESULTS
  Using 454 pyrosequencing, we obtained abundant high-quality ESTs from two spleen-complementary DNA libraries which were constructed from SGIV-infected (V) and PBS-injected fish (used as a control: C). A total of 407,027 and 421,141 ESTs were produced in control and SGIV infected libraries, respectively. Among the assembled ESTs, 9,616 (C) and 10,426 (V) ESTs were successfully matched against known genes in the NCBI non-redundant (nr) database with a cut-off E-value above 10-5. Gene ontology (GO) analysis indicated that "cell part", "cellular process" and "binding" represented the largest category. Among the 25 clusters of orthologous group (COG) categories, the cluster for "translation, ribosomal structure and biogenesis" represented the largest group in the control (185 ESTs) and infected (172 ESTs) libraries. Further KEGG analysis revealed that pathways, including cellular metabolism and intracellular immune signaling, existed in the control and infected libraries. Comparative expression analysis indicated that certain genes associated with mitogen-activated protein kinase (MAPK), chemokine, toll-like receptor and RIG-I signaling pathway were alternated in response to SGIV infection. Moreover, changes in the pattern of gene expression were validated by qRT-PCR, including cytokines, cytokine receptors, and transcription factors, apoptosis-associated genes, and interferon related genes.
  
  
  CONCLUSIONS
  This study provided abundant ESTs that could contribute greatly to disclosing novel genes in marine fish. Furthermore, the alterations of predicted gene expression patterns reflected possible responses of these fish to the virus infection. Taken together, our data not only provided new information for identification of novel genes from marine vertebrates, but also shed new light on the understanding of defense mechanisms of marine fish to viral pathogens.
authors.affiliation: Key Laboratory of Marine Bio-resources Sustainable Utilization, South China Sea Institute of Oceanology, Chinese Academy of Sciences, 164 West Xingang Road, Guangzhou, PR China.
authors.forename: Youhua
authors.initials: Y
authors.lastname: Huang
authors.forename: Xiaohong
authors.initials: X
authors.lastname: Huang
authors.forename: Yang
authors.initials: Y
authors.lastname: Yan
authors.forename: Jia
authors.initials: J
authors.lastname: Cai
authors.forename: Zhengliang
authors.initials: Z
authors.lastname: Ouyang
authors.forename: Huachun
authors.initials: H
authors.lastname: Cui
authors.forename: Peiran
authors.initials: P
authors.lastname: Wang
authors.forename: Qiwei
authors.initials: Q
authors.lastname: Qin
mesh_terms: D000818:Animals; D004266:DNA Virus Infections / Q000235:genetics / Q000662:veterinary*; D020224:Expressed Sequence Tags; D005393:Fish Diseases / Q000235:genetics* / Q000821:virology; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D015723:Gene Library; D017994:Iridovirus / Q000472:pathogenicity*; D010473:Perciformes / Q000235:genetics* / Q000821:virology; D017422:Sequence Analysis, DNA; D013154:Spleen / Q000378:metabolism* / Q000821:virology; D059467:Transcriptome
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