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.

Sex-lethal (Sxl) plays an important role in sex determination in insects. In this study, the full-length complementary DNA (cDNA) of Sxl of the Chinese mitten crab Eriocheir sinensis (EsSxl) was cloned. EsSxl is expressed during different developmental stages of embryos, and its expression level in the cleavage stage is lower than that in other stages of embryonic development, such as the original zoea stage when it reaches the highest level. The expression level of EsSxl in eight different tissues was investigated. EsSxl was expressed in seven examined tissues, excluding eyestalk, and the highest expression levels were observed in testis and hepatopancreas. EsSxl expression in testis was 13-fold higher than that in ovary. After induction by eyestalk ablation, changes in EsSxl expression were also tissue-specific, with EsSxl expression increasing by 2.6-fold and 11.5-fold in ovary and muscle, respectively, between 0 and 7 days after eyestalk ablation and decreasing by 2.0-fold in testis between 0 and 3 days after eyestalk ablation and by 265-fold in hepatopancreas between 0 and 7 days after eyestalk ablation. Two splice variants of EsSxl were identified, and the only difference between them was a 77-bp alternatively spliced intron that is transcripted in EsSxl1 and skipped in EsSxl2. Both variants were expressed in males and females, demonstrating the lack of a sex-specific expression pattern for Sxl in Chinese mitten crab.

A novel Eriocheir sinensis reovirus (EsRV) was identified using deep-sequencing techniques in crabs afflicted with trembling disease (TD). Near-full-length genome sequences of 12 segments of EsRV were obtained. The genome of EsRV will facilitate further studies on the causative agent of TD.