Investigation Of Sex Related Genes In Early Development Stage Of E.sinensis

Investigation of sex determination system in Eriocheir sinensis is important because of sex-dimorphism in its growth traits.However,little information about the sexrelated genes in embryonic and larval development stages were exposed.To obtain more information of sex determination in Chinese mitten crab,we combined the transcriptome in embryonic development stage in Eriocheir sinensis and the female specific marker to analysis the expression pattern of sex-related genes in early development stage in E.sinensis..Firstly,we performed the transcriptome analysis in embryonic development stage(fertilized egg stage,cleavage stage,blastula stage,gastrula stage and heartbeat stage)of Chinese mitten crab using next-generation sequencing technology.Thirty-one of 32,088 annotated unigenes were identified as sex-related genes including 16 genes involved in primary sex determination in model organisms and 8 genes of SOX family and 7 genes of DMRT gene family.Heatmap based on the RPKM value in five embryonic development stages indicated that these genes were clustered into two branches.Analysis of the differentially expressed 12 genes,including 3 genes of SOX family,3 genes of DMRT gene family and 6 genes involved in primary sex determination in model organisms,showed significantly difference between the first three periods(fertilized egg stage-cleavage stage-blastula stage)and the last two periods(gastrula stage-heartbeat stage)and all 12 genes were up-regulated after blastula stage.The 12 genes were regarded as the candidate genes of sex determination in E.sinensis.Secondly,we preformed the large-scale genome alignment between female and male in E.sinensis and obtained a female specific In Del marker-F23&R23,F23&R23 was mapped to scaffold8227 in female genome.We found three codon sequence: 8227-cds1,8227-cds2 and 8227-cds3 and performed the expression pattern of the three genes.Finally,we performed the expression pattern of the candidate sex determination genes in the Chinese mitten crabs which was identified sex by q RT-PCR.In conclution,we inferred that the sex determination might initiated after blastula stage and the ovaries development might initiated earlier than the testes development in E.sinensis.The study enriched the genetic resources of the E.sinensis and provided a background information for further investigation in sex determination of E.sinensis.