Discovery of Sex-Determining Genes in Portunus trituberculatus: A Comparison of Male and Female Transcriptomes During Early Developmental Stages

Abstract

Portunus trituberculatus is one of the main mariculture crabs of high economic value. To identify genes involved in sex determination, we first performed sex-specific transcriptome sequencing at six larval development stages using a DNA/RNA co-extraction method. A total of 907,952,938 and 828,774,880 reads were obtained from female and male crabs, respectively. 2,379 differentially expressed genes (DEGs) were found between females and males, and were mainly enriched in DNA replication, folate biosynthesis, and retinol metabolism pathways. Furthermore, transcription patterns of genes in the sex-determining region (SD) were analyzed based on the transcriptome data, and one Dmrt gene (PtDMY) was found to be exclusively expressed in males during early developmental stages. Notably, some known sex-related genes, including IAG, Dmrt11E, DmrtB1, and DmrtC2 were significantly down-regulated after knocking down PtDMY. Our results suggested that PtDMY is involved in sex determination and may be one of the key upstream regulators of the sex determination pathway. In addition, the massive volume of transcriptome data obtained in this study provided an important basis for the systematic study of sex determination mechanisms in P. trituberculatus.

CRISPR/Cas9-mediated gene mutation of EcIAG leads to sex reversal in the male ridgetail white prawn Exopalaemon carinicauda

“…The in-depth study on the sexual differentiation of the IAG gene provides a basis for the use of CRISPR/Cas9 gene editing technology to achieve sex manipulation in crustaceans. In addition to IAG, several genes homologous to Sxl, Tra-2, Dmrt and Dsx, which are involved in the insect sexual differentiation cascade, have been identified from many crustaceans (49)(50)(51)(52)(53). These genes share high sequence similarity with their homologs in insects and their expression profiles have been characterized in crustaceans.…”

Monosex Populations of the Giant Freshwater Prawn Macrobrachium rosenbergii—From a Pre-Molecular Start to the Next Generation Era

“…The availability of NGS technologies yielded novel molecular and genomic tools that were applied to several crustacean species [104]. Ample transcriptomic libraries of decapod crustaceans were studied, with several referring to sexually biased transcripts [27,28,[105][106][107][108], including the description of different stages in the life cycle of M. rosenbergii [109][110][111]. This is true also at the genomic level, with published genomes of decapod species, including the crayfish Procambarus virginalis [112] and C. quadricarinatus [113], the crabs E. sinensis [114,115], Birgus latro and Paralithodes camtschaticus [116], the ridgetail white prawn Exopalaemon carinicauda [117], the shrimp Neocaridina denticulata [118], L. vannamei [119], Marsupenaeus japonicus and Penaeus monodon [120], and the lobsters Panulirus ornatus [116] and Homarus americanus [121].…”

Comparative transcriptome analysis reveals the growth and development in larval stages of the swimming crab Portunus trituberculatus

“…At present, domestic and foreign research on swimming crab larvae is mainly focused on external factors, such as larval rearing (Dan et al, 2017), diseases (Muroga et al, 1989), morphology (Hamasaki et al, 2011;Dan et al, 2013), and environmental factors (Ren et al, 2017;Gao and Wang, 2021;Zhu et al, 2022). In recent years, genomics has been widely applied in studying crustaceans to elucidate the molecular mechanisms of different physiological processes (Li et al, 2014;Huerlimann et al, 2018;Zhao et al, 2021;Zhang et al, 2022). Compared to other sequencing methods, gene microarray technology can rapidly and efficiently obtain expression results for many genes at the transcriptional level, enabling genome-wide gene expression analysis (Bellin et al, 2009).…”