Sex-relevant genes in the embryo stage of Chinese soft-shelled turtles as revealed by RNA-Seq analysis

The Chinese soft-shelled turtle (Pelodiscus sinensis) is an economically important and rare freshwater aquaculture species in China. The male turtles have better economic benefits than females due to their faster growth speed. In order to explore the mechanism of sex differentiation and determination of P. sinensis, transcriptome analysis was carried out using embryo samples from the sex determination, sex differentiation and shelling stages, respectively. The gender types of embryos were identified by genotyping and histological analysis. In all, 277,230 and 273,859 genes were identified from embryos of male and female turtles, respectively. Cluster analysis of gene expression patterns exhibited trends for all differentially expressed genes (DEGs), which can be classified into three subclusters. In the sex determination, sex differentiation, and shelling stages 975, 43, and 300 DEGs, respectively, were identified by comparing the male and female groups. KEGG analysis was used to explore the function of the DEGs. Fgf9, foxl2, mex3b, sox7,tgfβ-3, wnt4, wnt7a, and wt1 were upregulated in ovary development stages and chfr, ampk, aldh2, dao, glt1d1, hvcn1, psmd9, srsf9 and ubc were upregulated in testis development stages, indicating that these genes play important roles in the sex development of P. sinensis. The results of this study provide preliminary insights into the sex determination and differentiation of Chinese soft-shelled turtles.