Jun Li

Spotted knifejaw (Oplegnathus punctatus), an economically important species in marine aquaculture, employs a unique sex determination mechanism based on a complex sex chromosome system (X₁X₁X₂X₂/X₁X₂Y). Males (2n = 47) possess one fewer chromosome than females (2n = 48), and their karyotype includes an unusually large neo-Y chromosome. Additionally, a pronounced sexual dimorphism in growth rate is observed, with males exhibiting a faster growth rate than females. In this study, we conducted a comprehensive whole-genome scan, which initially revealed structural variations in the anti-inflammatory itih4 gene between male and female O. punctatus. Additionally, we designed a pair of primers to detect DNA sequence variations within the itih4a/itih4b gene. These variations are located in the intergenic region of the fusion Y chromosome in male O. punctatus, compared to the homologous X chromosome in females. In females without DNA insertions in the itih4a/itih4b intergenic region, a single band of 351 bp is amplified. By contrast, in males with DNA insertions, two bands are amplified (755 bp and 351 bp). The 755 bp band specifically indicates the presence of a DNA insertion in the itih4a/itih4b intergenic region on the Y chromosome, associated with male-specific genetic traits. Our study will facilitate the rapid identification of the genetic sex of both male and female O. punctatus individuals.

Recirculating aquaculture systems (RAS) have promising applications in aquaculture. Feed is recognized as a major source of input to the RAS, and feeding frequency will not only impact the performance of turbot, but will also impact the quality of the cultured water. In order to rationally manage feeding and reduce aquaculture pollution, this study investigated the effects of feeding frequency on the performance of turbot (Scophthalmus maximus), nitrogen removal (ammonia and nitrite) characteristics and microbial communities in biofilters. The experiment was designed with three treatment groups, which were categorized into feeding once/day (FF1), feeding twice/day (FF2) and feeding three times/day (FF3) for 30 days. The results indicated that weight gain rate (WGR) and specific growth rate (SGR) significantly increased (p < 0.05) in the FF2 group and FF3 group compared with the FF1 group. The feed conversion ratio (FCR) was significantly lower (p < 0.05) in the FF2 group and FF3 group than in the FF1 group. There was no significant change in condition factor (CF). Ammonia and nitrite concentration decreased and water quality fluctuated less as the feeding frequency increased. FF2 showed the highest ammonia and nitrite removal rates. Feeding frequency did not significantly affect biofilter alpha diversity, but significantly altered beta diversity. PICRUSt functional prediction analysis revealed that the relative abundance of functional genes for nitrogen metabolism (amoA, amoB, amoC, hao, nxrA and nxrB) was highest in FF2. Therefore, feeding frequency of twice/day not only benefits the performance of turbot but also stabilizes the water environment and improves the removal of ammonia nitrogen and nitrite in RAS. These results provide theoretical and practical basis for further water improvement by seawater RAS.

Spermatogenesis, the process of male germ cell development, is tightly regulated by the hypothalamic–pituitary–testis (HPT) axis in seasonally breeding teleosts. Despite its importance, our understanding of how the brain and male germ cells coordinate key transitions—such as testis initiation and maturation—remains limited, particularly in species with distinct seasonal reproductive cycles. Black rockfish (Sebastes schlegelii), a marine viviparous teleost, exhibits a prolonged testis quiescent phase lasting three-quarters of the year, with testis initiation occurring in September and maturation concluding in November and December. The mechanisms underlying these transitions are poorly characterized, leaving a critical gap in our knowledge of seasonal spermatogenesis and its regulation. Addressing this gap is crucial for advancing artificial breeding technologies, which could significantly benefit the aquaculture industry. RNA-seq was used to explore the gene regulatory networks involved in testis initiation in S. schlegelii. Transcriptomic analyses of brain and testis were conducted across key developmental phases. In the brain, upregulated genes were notably involved in neuroactive ligand–receptor interactions, whereas in the testis, differentially expressed genes were linked to cell cycle processes and ATP-dependent chromatin remodeling. Our findings reveal the molecular mechanisms underlying testis initiation in S. schlegelii, providing omics evidence for the role of the HPT axis in regulating this process. By elucidating the gene regulatory networks of the brain and testis during critical transitions, this study advances our understanding of spermatogenesis in seasonally breeding teleosts. These insights pave the way for developing year-round artificial breeding technologies, contributing to the sustainable management of commercially valuable fish species.

The fish species Oplegnathus fasciatus exhibits an X₁X₁X₂X₂/X₁X₂Y sex determination mechanism. This species holds considerable economic value and displays pronounced sexual dimorphism in growth. Therefore, the development of a rapid and accurate method for sex identification is critical to enhancing breeding efficiency and maximizing production value. Using third-generation PacBio whole-genome sequencing, we identified a homologous region in the samd3/elf3 intergenic region of the X and Y chromosomes of O. fasciatus. Analysis of the whole-genome sequence revealed a large DNA insertion marker fragment within this region. Using specifically designed primers, two bands of 390 bp and 1008 bp were successfully amplified in males, whereas only a single 390 bp band was detected in females. This marker can be easily distinguished by agarose gel electrophoresis, greatly enhancing the efficiency and accuracy of sex identification. This study not only expands the molecular marker system for sex identification of O. fasciatus but also offers a valuable methodological reference for sex identification in other economically important fish species. These findings have significant implications for germplasm improvement and efficient selection in aquaculture.

Sex determination mechanisms vary significantly across different chromosomal systems and evolutionary contexts. Nonetheless, the regulatory framework governing the multi-sex chromosome system (X1X1X2X2/X1X2Y) remains enigmatic. Through an examination of sex-related genes (dmrt1, hsd11b2, amh, sox9a, sox9b, foxl2, cyp19a), hormonal influences (E2, 11-KT), and histological analyses of gonadal development, we demonstrate that the critical period for sexual differentiation occurs between 35 to 60 days post-hatching (dph). Our multi-omics analysis identified amhr2 as a candidate sex-determining gene, revealing that the males possess three distinct amhr2 transcripts (amhr2ay, amhr2by, amhr2cy), whereas females express only one (amhr2a). In situ hybridization assays demonstrated that amhr2 is predominantly localized to primary spermatocyte and Sertoli cells of male testes. Notably, the specific mRNA expression of amhr2 is significantly enriched in amhr2cy, whose extracellular domain exhibits the highest binding affinity for Amh protein, with sexual expression differences manifesting as early as 5 dph. The outcomes of amhr2 interference (RNAi) experiments indicate that amhr2 knockdown leads to a reduction in the expression of male-related gene (dmrt1, amh, sox9a, sox9b), androgen synthesis genes (hsd11b2, cyp11a), and female-related genes (wnt4, foxl2, cyp19a, cyp19b). Conversely, overexpression of amhr2 yielded contrasting results. Our research supports the role of amhr2 as a pivotal candidate sex-determining gene. Furthermore, the dosage effect of amhr2, reflected in transcript abundance, mRNA expression levels, and binding efficacy, serves as a fundamental mechanism driving male differentiation and regulatory processes in Spotted knifejaw.