Screening and characterization of sex-specific markers by NGS sequencing in Spinibarbus hollandi with implication of XY sex determination system

Sex-specific markers are critical for understanding sex determination mechanism and development of unisexual breeding in fish. Spinibarbus hollandi is an important commercial aquaculture species in southeastern China. Here, through whole genome sequencing of three female and six male individuals of S. hollandi, we first screened out a total of 88 candidate male-specific sequences. After PCR validation, three male-specific markers were proved to be able to realize male-specific amplification in S. hollandi. Subsequently, the three markers were also examined in another 24 female and 24 male individuals of S. hollandi, and the accuracy was 100%. Moreover, several male-specific genes that participate in sex determination process were identified by NR annotation, which might act as sex determining genes in S. hollandi. In general, the male-specific markers suggested an XY sex determination system in S. hollandi and would contribute a lot to understanding of sex determination mecha-nisms and development of unisexual breeding.