Prediction of suitable habitat shifts and assessment of ecological niche overlaps for three Tridentiger species with intertidal and subtidal characteristics under future climate changes.

To show how dramatic global climate change affects marine ecosystem species in different habitats. We used a joint species distribution model (SDM) and an ecological niche model (ENM) to investigate the suitable habitat shifts and ecological niche overlaps of the Tridentiger fishes. In the present study, the SDM results showed that 5 hotspots were identified for T. trigonocephalus and T. barbatus, and 4 hotspots for T. bifasciatus. The study on center-of-mass transfer revealed notable reductions in the habitual range of the three Tridentiger species with future climate change and no significant bipolar shifts in the center of mass. The ENM results indicated that T. trigonocephalus and T. barbatus exhibited the greatest ecological niche overlap with Schoener's D (D) and Hellinger-based I (I) values of 0.4719 and 0.7690, respectively. Both SDM and ENM results have suggested that T. trigonocephalus occupied a wider distribution and greater adaptability to future climate change. This study sought to measure the variations in the effects of global climate change on marine species in different habitats. Our study first found that intertidal species with specific life histories may be more resilient to environmental change.

Theoretical calculations and photoluminescence properties of Sr4Al14O25:Mn4+ red phosphors doped with Li.

The absence of high-efficiency oxide red phosphors restricts the development of high-performance solid-state lighting. In this work, a series of Li+ doped Sr4Al14O25:Mn4+ (SAO-Li+) red phosphors were prepared. Theoretical calculation results indicate that Li+ is inclined to occupy the gap 2 position. The low concentration Li+ gap doping has almost no influence on the Sr4Al14O25 structure, and a 0.4 mol Li+-doped sample exhibits a pure phase with regular morphology. With increasing Li+ doping content, the luminescence intensities of phosphors increase first and then decrease. 0.4 mol is found to be the optimal concentration. The fluorescence lifetime continues to decrease with the increase in the Li+ doping content and a mutation occurs at 0.5 mol Li+. Phosphor doping with Li+ can improve the thermal quenching resistance. The WLED device encapsulated with SAO-0.4Li and YAG:Ce3+ phosphors prepared showed a correlated color temperature of 4667 K, a color rendering index of 82, and a light efficiency of 137.34 lm W-1 at a driving current of 20 mA. The above results indicate that the use of a SAO-0.4Li+ phosphor is expected for application in warm WLEDs.

First observation of the mating, egg-laying and hatching behaviour of a captive female Kong skate, Okamejei kenojei (Müller & Henle, 1841).

This study provides a novel record of the reproductive behaviour of the Kong skate (Okamejei kenojei) in captivity. These skates were found to mate and deposit eggs at a temperature of 16.5 ± 0.5°C. The results showed that 76.13% of the eggs possessed one yolk, 0.77% of the eggs possessed two yolks and 23.11% of the eggs had no yolk (N = 1043). The deposition of non-yolk and double-yolk eggs was random. A total of 100 eggs were collected. After nearly 92 ± 5 days of incubation, 28 eggs failed to hatch, and 72 skates were successfully hatched with a female-to-male ratio of 1:1 (P > 0.05). The results enrich our knowledge of the reproduction in cartilaginous fishes and can inform management and conservation strategies for this species.

Chromosome-Level Genome Reveals the Origin of Neo-Y Chromosome in the Male Barred Knifejaw Oplegnathus fasciatus.

The barred knifejaw, Oplegnathus fasciatus, is characterized by an X1X2Y system with a neo-Y chromosome for males. Here, a chromosome-level genome was assembled to investigate the origin of neo-Y chromosome to the male O. fasciatus. Twenty-three chromosomes corresponding to the male karyotypes were scaffolded to 762-Mb genome with a contig N50 length of 2.18 Mb. A large neo-Y chromosome (Ch9) in the male O. fasciatus genome was also assembled and exhibited high identity to those of the female chromosomes Ch8 and Ch10. Chromosome rearrangements events were detected in the neo-chromosome Ch9. Our results suggested that a centric fusion of acrocentric chromosomes Ch8 and Ch10 should be responsible for the formation of the X1X2Y system. The high-quality genome will not only provide a solid foundation for further sex-determining mechanism research in the X1X2Y system but also facilitate the artificial breeding aiming to improve the yield and disease resistance for Oplegnathus.

Genome sequence of the barred knifejaw Oplegnathus fasciatus (Temminck & Schlegel, 1844): the first chromosome-level draft genome in the family Oplegnathidae.

BACKGROUND: The barred knifejaw (Oplegnathus fasciatus), a member of the Oplegnathidae family of the Centrarchiformes, is a commercially important rocky reef fish native to East Asia. Oplegnathus fasciatus has become an important fishery resource for offshore cage aquaculture and fish stocking of marine ranching in China, Japan, and Korea. Recently, sexual dimorphism in growth with neo-sex chromosome and widespread biotic diseases in O. fasciatus have been increasing concern in the industry. However, adequate genome resources for gaining insight into sex-determining mechanisms and establishing genetically resistant breeding systems for O. fasciatus are lacking. Here, we analyzed the entire genome of a female O. fasciatus fish using long-read sequencing and Hi-C data to generate chromosome-length scaffolds and a highly contiguous genome assembly. FINDINGS: We assembled the O. fasciatus genome with a total of 245.0 Gb of raw reads that were generated using both Pacific Bioscience (PacBio) Sequel and Illumina HiSeq 2000 platforms. The final draft genome assembly was approximately 778.7 Mb, which reached a high level of continuity with a contig N50 of 2.1 Mb. The genome size was consistent with the estimated genome size (777.5 Mb) based on k-mer analysis. We combined Hi-C data with a draft genome assembly to generate chromosome-length scaffolds. Twenty-four scaffolds corresponding to the 24 chromosomes were assembled to a final size of 768.8 Mb with a contig N50 of 2.1 Mb and a scaffold N50 of 33.5 Mb using 1,372 contigs. The identified repeat sequences accounted for 33.9% of the entire genome, and 24 003 protein-coding genes with an average of 10.1 exons per gene were annotated using de novo methods, with RNA sequencing data and homologies to other teleosts. According to phylogenetic analysis using protein-coding genes, O. fasciatus is closely related to Larimichthys crocea, with O. fasciatus diverging from their common ancestor approximately 70.5-88.5 million years ago. CONCLUSIONS: We generated a high-quality draft genome for O. fasciatus using long-read PacBio sequencing technology, which represents the first chromosome-level reference genome for Oplegnathidae species. Assembly of this genome assists research into fish sex-determining mechanisms and can serve as a resource for accelerating genome-assisted improvements in resistant breeding systems.

Spermatogonial stem cells differentiation and testicular lobules formation in a seasonal breeding teleost: The evidence from the heat-induced masculinization of genetically female Japanese flounder (Paralichthys olivaceus).

Germ cells undergo a series of cellular changes including differentiation, mitosis, meiosis and maturation, and eventually develop into a large number of functional gametes. The available data regarding teleost gametogenesis in seasonal batch spawners is limited. In this study, we investigated spermatogenesis with special attention on spermatogonia differentiation using heat-induced masculine juveniles of genetically female Japanese flounder. Meanwhile, the Nanos2 expression had been detected by immunohistochemistry for analysis of spermatogonial stem cells (SSCs) distribution. Spermatogonia began mitosis at 35 dph, and basement membrane firstly appeared and gradually surrounded and separated the spermatogonia (type A) into single and paired status. At this period, the spermatogonia continuously maintained mitotic proliferation. As a result, the number of spermatogonia including isolated and clusters (2-8 spermatogonia) significantly increased in the presumptive testes. From 85 dph to 120 dph, with the mitosis of spermatogonia, germline acinar-clusters formed. In the clusters, type A spermatogonia differentiated into type B, and multi-spermatogonia surrounded by several sertoli cell formed cysts，which represented the formation of lobular precursors. After that, type B spermatogonia began meiosis, which indicated the initiation of spermatogenesis. In adult testes, most type A spermatogonia distributed in the peripheral region and a few clung to a basement membrane in the internal germinal epithelium. Various spermatogenic cysts with germ cells in different developmental stages existed in a testicular lobules, moreover the germ cells in earlier stages resided in the distal termini, and the advanced stages were adjacent to the spermatic duct of testes. Therefore, the testes of Japanese flounder belonged to an intermediated distribution of SSCs, which might contribute greatly to multiple spermiation during breeding season. These findings would improve the understanding the mechanisms of SSCs differentiation and testicular development, and may be of great value in future studies of the spermatogenesis regulation.

Germ Cell Migration, Proliferation and Differentiation during Gonadal Morphogenesis in All-Female Japanese Flounder (Paralichthys Olivaceus).

In this study, all genetic female (XX) broods of Japanese flounder were produced artificially by mating the females with sex-reversed males. The proliferation and migration of primordial germ cells (PGCs), formation of ovary and oogenesis were described in detail. After hatching, around 20 individual PGCs migrated from the lateral to the dorsal of trunk region. At 15 days posthatching (dph), a part of PGCs were covered by a single layer somatic cells and formed the genital ridge. By 22 dph, the elongated gonadal primordia appeared under the ventral kidney, where the PGCs were totally enclosed by somatic cells. During the process of migration, PGCs were presumed to be mitotically inactive. From 63 to 73 dph, somatic cells rearrangement resulted in the formation of a narrow crevice, which became deeper and formed ovarian lumen. However, at 52 dph, dramatic mitotic proliferation of germ cell occurred and germline nest formed before the appearance of ovarian lumen. The onset of intensive germ cell proliferation and appearance of cell nests could be accepted as a criterion of initial ovarian differentiation. Then germ cells and somatic epithelial cells were gradually delimited by basement membrane and formed the germinal epithelium. In this period, results from in situ hybridization revealed that the early forkhead box L2 (pofoxl2) was expressed in somatic cells and oocytes in primary growth, which indicated the prefollicle cells formed. Then oogonia or oocytes, follicle cells, basement membrane, and theca cells composed a follicle complex. Finally, oocytes underwent meiosis and developed into to mature eggs. Anat Rec, 301:727-741, 2018. © 2017 Wiley Periodicals, Inc.

The expression pattern of hsp70 plays a critical role in thermal tolerance of marine demersal fish: Multilevel responses of Paralichthys olivaceus and its hybrids (P. olivaceus ♀ × P. dentatus ♂) to chronic and acute heat stress.

Ocean warming has multifaceted impacts on marine organisms. This study investigated the different responses of Paralichthys olivaceus and the hybrids (P. olivaceus ♀ × P. dentatus ♂) to chronic and acute heat stress. By comparing their survival, behavioural and histological changes, we found that the hybrids possess a better thermal tolerance with a higher cumulative survival rate (CSR), relatively fewer behavioural changes and less gill damage. Moreover, we analysed the relationship between thermal tolerance and the hsp70 expression pattern and found that thermal tolerant species (the hybrids) exhibited higher threshold induction temperature, shorter durations, stronger magnitudes and a delay in hsp70 expression. We speculated that the expression mode of hsp70, rather than itself, plays a critical role in thermal tolerance. These findings would improve the understanding of hsp70 in future marine climate research and help clarify the profound effects of rising temperature on marine demersal fishes.

Pronounced population genetic differentiation in the rock bream Oplegnathus fasciatus inferred from mitochondrial DNA sequences.

The population genetic structure of the rock bream (Oplegnathus fasciatus) along the coastal waters of China was estimated based on three mtDNA fragments (D-loop, COI, and Cytb). A total of 112 polymorphic sites were checked, which defined 63 haplotypes. A pattern with high levels of haplotype diversity (hCOI = 0.886 ± 0.034, hCytb = 0.874 ± 0.023) and low levels of nucleotide diversity (лCOI = 0.009 ± 0.005, лCytb = 0.006 ± 0.003) was detected based on the COI and Cytb fragments, and high levels of genetic diversity (hD-loop = 0.995 ± 0.007, лD-loop = 0.021 ± 0.011) were detected from the mtDNA D-loop. The population genetic diversity of O. fasciatus in south China was significantly higher than those of north China. Three genealogical clades were checked in the O. fasciatus populations based on the NJ and MST analyses of mtDNA COI gene sequence, and the genetic distances among the clades ranged from 0.018 to 0.025. Significant population genetic differentiation was also checked based on the Fst (0.331, p = 0.000) and exact p (0.000) test analyses. No significant population differentiations were checked based on mtDNA D-loop and Cytb fragments. Using a variety of phylogenetic methods, coalescent reasoning, and molecular dating interpreted in conjunction with paleoclimatic and physiographic evidences, we inferred that the genetic make-up of extant populations of O. fasciatus was shaped by Pleistocene environmental impacts on the historical demography of this species. Coalescent analyses (neutrality tests, mismatch distribution analysis, and Bayesian skyline analyses) showed that the species along coastline of China has experienced population expansions originated in its most recent history at about 169-175 kya before present.

The dnd RNA Identifies Germ Cell Origin and Migration in Olive Flounder (Paralichthys olivaceus).

The present study obtained a germ cell-specific marker dead end (dnd) in olive flounder (Paralichthys olivaceus) named Podnd. The tissue-specific expressions of Podnd transcripts were present in testis and ovary but were not detectable in other somatic tissues detected. SISH showed that Podnd expressed only in germ cells at different developmental stages but not in surrounding somatic cells. The expression of Podnd during embryonic development at 16 different stages revealed that the relative expression of Podnd transcript fluctuated at a high level in the cleavage stages, gradually decreased through subsequent development, and reached the lowest at late gastrula stage till it was nearly undetectable. The Podnd transcripts localization and migration were similar to zebrafish. Further research on the specification migration mechanism of PGCs and the role of germ cell during gonadal development in olive flounder would improve our understanding of germline development.

Maternally derived trypsin may have multiple functions in the early development of turbot (Scopthalmus maximus).

Trypsin is an important serine protease that is considered to be involved in digestion of protein in teleost fish. Nevertheless, studies on trypsin/trypsinogen in fish embryos are very limited. In this study, the trypsinogen of turbot (Scophthalmus maximus) (tTG) was identified and the expression patterns and activity of trypsinogen/trypsin were investigated. The results showed that the tTG mRNA was evenly distributed in the oocytes and was also expressed along the yolk periphery in early embryos. At later embryo stages and 1 days after hatching (dph), the tTG mRNA concentrated at the alimentary tract and head. Quantitative expression analysis showed that the tTG transcripts decreased after fertilization until the gastrula stage, then increased with the embryo and larvae development. This result was also confirmed by the specific activity analysis of trypsin and in-situ-hybridization (ISH). All of the results indicated that tTG in early embryo stages was maternally derived and expressed by itself after gastrula stages. Additionally, location of tTG mRNA in embryos and larvae was investigated; we considered that trypsin may have multiple functions during the embryo development process. Based on our results regarding trypsinogen in embryos and early development, we concluded that the trypsin/trypsinogen in turbot embryos was inherited from a maternal source and we suggested that trypsin in early development has multiple functions in the process of development.

Effect of the addition of six antioxidants on sperm motility, membrane integrity and mitochondrial function in red seabream (Pagrus major) sperm cryopreservation.

The present study was to evaluate the effects of six antioxidants on frozen-thawed sperm motility, viability, membrane integrity and mitochondrial function in red seabream (Pagrus major) by computer-assisted sperm analysis system and flow cytometry, respectively. All the parameters tested in this study were determined using one-way ANOVA and identified using the SNK test (P < 0.05). The results demonstrated that on the first day, the highest motility and longevity occurred in 100 mM trehalose (78.34 ± 3.41%, 29 ± 4.00 days) and 50 mM taurine (77.46 ± 1.54%, 29.33 ± 4.04 days), followed by 25 mM vitamin C (79.03 ± 5.37 %, 17 ± 1.00 days), 25 mM vitamin E (69.64 ± 1.64%, 27.67 ± 1.53 days) and 25 mM vitamin A (78.89 ± 2.81%, 9.33 ± 1.53 days), which were all higher than frozen-thawed sperm without antioxidant (control) (66.80 ± 5.55, 5.67 ± 1.15 days). Especially, the percentages of class A sperm with the addition of 100 mM trehalose (40.39 ± 5.20%) and 50 mM taurine (37.78 ± 3.22%) were significantly improved compared to the control (19.63 ± 5.44%). The viability of all groups on the third and sixth day showed a similar trend. Moreover, during the 4 °C storage process, the decrease of frozen-thawed sperm motility was closely associated with the decrease in membrane integrity and mitochondrial function. In conclusion, the present study indicated that antioxidant (100 mM trehalose and 50 mM taurine) provided the most pronounced protective effect in improving frozen-thawed quality of red seabream sperm. The addition of antioxidant may be capable of scavenging the ROS generated during the cryopreservation process and 4 °C storage.

Histological and enzymatic responses of Japanese flounder (Paralichthys olivaceus) and its hybrids (P. olivaceus ♀ × P. dentatus ♂) to chronic heat stress.

This study investigated the effects of long-term heat exposure on Japanese flounder (Paralichthys olivaceus) and its hybrids (P. olivaceus ♀ × summer flounder Paralichthys dentatus ♂). From 24 ± 0.5°C, temperature was increased by 1 ± 0.5°C in a day and was kept at that temperature for 5 days before next rise. Cumulative survival rate (CSR), cumulative survival rate under different temperature (CSR-T), histological alteration, and related enzyme activities were investigated. In P. olivaceus, mass mortality occurred at 29 and 32 °C (the CSR-T dropped to 42.39%), and serious gill damages appeared at 30 and 32°C. Meanwhile, the activities of superoxide dismutase (SOD), catalase (CAT), lysozyme (LZM), and pyruvate kinase (PK) declined around 29 and 32°C (except for CAT). In comparison with P. olivaceus, the CSR of the hybrids was higher, the gill kept a better structural integrity, and the activities of SOD, CAT, LZM, and PK showed tiny fluctuations. The results suggested that during the process of chronic heat stress, P. olivaceus seemed to be more sensitive to 29 and 32°C, and the manifestations in survival, histology, and enzyme activity were generally consistent. For the hybrids, the comparatively insensitivity to high temperature might imply its better heat tolerance.

Pepsinogen A and C genes in turbot (Scophthalmus maximus): characterization and expression in early development.

We characterized the expression patterns of pepsinogen A (tPGA) and pepsinogen C (tPGC) in turbot (Scophthalmus maximus). Quantitative expression analysis showed that tPGC was preferentially expressed in early developmental stages, and that the tPGA mRNA expression level was higher in adult fish. Full-length cDNA constructs of tPGA and tPGC were 1307 bp (from which 377 amino acids were deduced); and 1430 bp (from which 385 amino acids were deduced), respectively. The deduced proteins of tPGA and tPGC possessed signal peptides of 17 amino acids and 20 amino acids respectively. The initial transcripts of tPGA and tPGC were detected at 22 days post hatching (dph), well after the formation of gastric glands (16 dph). This suggested that the morphologic development of gastric glands was not synchronous with their functional development. In addition, tPGA and tPGC mRNAs were also expressed in muscle and ovary at much lower levels than in stomach and esophagus. The distribution of tPGA and tPGC in the turbot was investigated using in-situ hybridization, and tPGA and tPGC were first detected in the esophagus and cardiac region of the stomach, and then throughout the stomach.

Germ line specific expression of a vasa homologue gene in turbot (Scophthalmus maximus): evidence for vasa localization at cleavage furrows in euteleostei.

Specification of primordial germ cells during early embryogenesis is a critical biological issue in reproduction and development. Yet, little is known in marine economic fish species. Vasa, a component of germ plasm, is the most-documented germ cell marker in teleosts. We isolated a full-length vasa cDNA (Smvas) from turbot (Scophthalmus maximus), a marine Euteleostei species, and investigated its expression patterns by RT-PCR and in situ hybridization during embryogenesis and gametogenesis to identify the germ cell lineage in this species. The deduced amino acid sequence of the isolated cDNA shared typical characteristics of Vasa protein and high identity to Vasa homologues in medaka (76.9%) and zebrafish (68.5%). The Smvas transcripts were exclusively detected in germ cells of testis and ovary, and exhibited an interesting dynamic localization pattern during oogenesis. The distribution pattern of Smvas during embyogenesis in this Euteleostei closely resembled the pattern observed in zebrafish (belonging to Osteriophysans) rather than medaka (belonging to Euteleostei). Thus, it is concluded that Smvas isolated in this study is a germ cell specific molecular marker in turbot. Furthermore, we hypothesize that Euteleostei could localize vasa mRNA by a special mode. The results not only facilitate the germ cell manipulation of the turbot, but also improve our understanding of germline development and evolution of vasa localization in teleost.

Ontogeny of the digestive tract and enzymes in rock bream Oplegnathus fasciatus (Temminck et Schlegel 1844) larvae.

Histological development of the digestive tract and specific activities of three digestive enzymes (trypsin, alkaline phosphatase, and pepsin) were studied in rock bream Oplegnathus fasciatus from hatching to 50 days after hatching (DAH). At hatching, the digestive tract appeared as an undifferentiated straight tube and differentiated into the buccopharynx, esophagus, stomach, intestine, and rectum at mouth opening by 3 DAH. The taste bud and mandibular teeth were present in the buccopharyx at 8 DAH. The goblet cells appeared in the esophagus at 8 DAH and in the buccopharyx at 9 DAH. The stomach anlage was formed at 2 DAH and developed into cardia, fundus, and pylorus at 14 DAH. The gastric glands were visible at 16 DAH, and the pepsin was firstly detected on 22 DAH. At 2 DAH, the intestinal valve appeared and divided the intestine into anterior intestine (AI) and posterior intestine (PI). The rectum was differentiated from the PI at 3 DAH. The supranuclear vacuoles were visible in the rectum by 6 DAH, and the lipid inclusions were present in the AI at 8 DAH. The alkaline phosphatase was detected at 1 DAH, and the increase in its activity indicated the maturation of the intestine after 40 DAH. The hepatocytes and pancreatic cells were differentiated from the blast cells at 2 DAH, and the acidophilic zymogen granules in the exocrine pancreas were observed simultaneously. The trypsin was detected by 1 DAH and increased to the maximum at 19 DAH, followed by a decrease as the stomach became functional.

Population genetic structure of the point-head flounder, Cleisthenes herzensteini, in the Northwestern Pacific.

Intraspecific phylogenies can provide useful insights into how populations have been shaped by historical and contemporary processes. To determine the population genetic structure and the demographic and colonization history of Cleisthenes herzensteini in the Northwestern Pacific, one hundred and twenty-one individuals were sampled from six localities along the coastal regions of Japan and the Yellow Sea of China. Mitochondrial DNA variation was analyzed using DNA sequence data from the 5' end of control region. High levels of haplotype diversity (>0.96) were found for all populations, indicating a high level of genetic diversity. No pattern of isolation by distance was detected among the population differentiation throughout the examined range. Analyses of molecular variance (AMOVA) and the conventional population statistic Fst revealed no significant population genetic structure among populations. According to the exact test of differentiation among populations, the null hypothesis that C. herzensteini within the examined range constituted a non-differential mtDNA gene pool was accepted. The demographic history of C. herzensteini was examined using neutrality test and mismatch distribution analyses and results indicated Pleistocene population expansion (about 94-376 kya) in the species, which was consistent with the inference result of nested clade phylogeographical analysis (NCPA) showing contiguous range expansion for C. herzensteini. The lack of phylogeographical structure for the species may reflect a recent range expansion after the glacial maximum and insufficient time to attain migration-drift equilibrium.