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In-depth comparative transcriptome analysis of intestines of red swamp crayfish, Procambarus clarkii, infected with WSSV.

ABSTRACT: Crayfish has become one of the most important farmed aquatic species in China due to its excellent disease resistance against bacteria and viruses. However, the antiviral mechanism of crayfish is still not very clear. In the present study, many high-quality sequence reads from crayfish intestine were obtained using Illumina-based transcriptome sequencing. For the normal group (GN), 44,600,142 high-quality clean reads were randomly assembled to produce 125,394 contigs. For the WSSV-challenged group (GW), 47,790,746 high-quality clean reads were randomly assembled to produce 148,983 contigs. After GO annotation, 39,482 unigenes were annotated into three ontologies: biological processes, cellular components, and molecular functions. In addition, 15,959 unigenes were mapped to 25 different COG categories. Moreover, 7,000 DEGs were screened out after a comparative analysis between the GN and GW samples, which were mapped into 250 KEGG pathways. Among these pathways, 36 were obviously changed (P-values < 0.05) and 28 pathways were extremely significantly changed (P-values < 0.01). Finally, five key DEGs involved in the JAK-STAT signaling pathway were chosen for qRT-PCR. The results showed that these five DEGs were obviously up-regulated at 36 h post WSSV infection in crayfish intestine. These results provide new insight into crayfish antiviral immunity mechanisms.

SUBMITTER: Du Z

PROVIDER: S-EPMC4901281 | biostudies-other | 2016

REPOSITORIES: biostudies-other

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Project description:The red swamp crayfish Procambarus clarkii is a highly adaptable, tolerant, and fecund freshwater crayfish that inhabits a wide range of aquatic environments. It is an important crustacean model organism that is used in many research fields, including animal behavior, environmental stress and toxicity, and studies of viral infection. Despite its widespread use, knowledge of the crayfish genome is very limited and insufficient for meaningful research. This is the use of next-generation sequencing techniques to analyze the crayfish transcriptome. A total of 324.97 million raw reads of 100 base pairs were generated, and a total of 88,463 transcripts were assembled de novo using Trinity software, producing 55,278 non-redundant transcripts. Comparison of digital gene expression between four different tissues revealed differentially expressed genes, in which more overexpressed genes were found in the hepatopancreas than in other tissues, and more underexpressed genes were found in the testis and the ovary than in other tissues. Gene ontology (GO) and KEGG enrichment analysis of differentially expressed genes revealed that metabolite- and immune-related pathway genes were enriched in the hepatopancreas, and DNA replication-related pathway genes were enriched in the ovary and the testis, which is consistent with the important role of the hepatopancreas in metabolism, immunity, and the stress response, and with that of the ovary and the testis in reproduction. It was also found that 14 vitellogenin transcripts were highly expressed specifically in the hepatopancreas, and 6 transcripts were highly expressed specifically in the ovary, but no vitellogenin transcripts were highly expressed in both the hepatopancreas and the ovary. These results provide new insight into the role of vitellogenin in crustaceans. In addition, 243,764 SNP sites and 43,205 microsatellite sequences were identified in the sequencing data. We believe that our results provide an important genome resource for the crayfish.

Dataset Information

In-depth comparative transcriptome analysis of intestines of red swamp crayfish, Procambarus clarkii, infected with WSSV.

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