Black rockfish(Sebastes schlegelii)has evolved viviparous reproduction mode.Different from the oviparous fishes,the spermatozoa of black rockfish are stored on the surface of ovarian follicle for over four months unti...Black rockfish(Sebastes schlegelii)has evolved viviparous reproduction mode.Different from the oviparous fishes,the spermatozoa of black rockfish are stored on the surface of ovarian follicle for over four months until the egg mature in ovary,while the adaptive evolution of fertilization-related genes remains to be studied.In the present study,based on the genome and transcriptome information of black rockfish,a total of 10 hyaluronidase genes were identified by phylogenetic and sequence analyses,including hyal1,hyal2,hyal3,hyal4,hyal6 and spam1a,spam1b,spam1c and spam1d.The spam1 subfamilies,including spam1a,spam1b,spam1c and spam1d,were significantly expanded compared to other species.The adaptive evolution of hyaluronidase was further investigated by selection pressure analysis of branch model and branch site model.The results showed that only spam1 subfamily was positively selected with a large number of positive selection sites,and the evolution rate was significantly higher thanthose of other teleosts.Two positively selected sites,LYS-171 and GLY-164,were located in the neutral hyaluronidase activity domain,which was pivotal for SPAM1 to participate in enzymatic hydrolysis of the extracellular hyaluronic acid matrix and mediate acrosome reaction in mammals.Furthermore,spam1 was mainly expressed in spermatocytes,secondary spermatocytes,and spermatids of testis,whereas it was not found in metamorphic sperm and mature sperm based on the spatiotemporal expression analysis.All results indicated that spam1 might originate from the testis and might be an adaptation of viviparous trait in black rockfish.展开更多
Mitogen-activated protein kinases(MAPKs)play pivotal roles in response to environmental stresses and bacterial infections.Compared with those in the higher vertebrates,studies of mapk gene family are still limited in ...Mitogen-activated protein kinases(MAPKs)play pivotal roles in response to environmental stresses and bacterial infections.Compared with those in the higher vertebrates,studies of mapk gene family are still limited in teleost.Identification,characterization,classification,and expression profiling of totally 15 mapk genes in black rockfish(Sebastes schlegelii)were conducted.Phylogenetic relationships show that these mapk genes could be divided into extracellular signal-regulated kinase(ERK),c-Jun N-terminal kinase(JNK),and p38 sub-families.In addition,gene structures,syntenic analysis,and selective pressure analysis are performed to confirm their annotations.Results of selective pressure analysis indicate that mapk1,mapk3,mapk7,mapk10,mapk11,and mapk12 underwent significantly-positive selections,while the others genes such as mapk4,mapk6,mapk15,mapk8a,mapk8b,mapk9,mapk13,mapk14a,and mapk14b were under purifying selections.Moreover,results of qRT-PCR indicate that mapk genes in 8 healthy tissues displayed different expression patterns.The expression patterns of several mapk genes including mapk12,mapk13,mapk14a,mapk14b,and mapk15 were significantly changed in mucosal tissues after Edwardsiella piscicida infection.This study demonstrates that mapk genes in black rockfish play vital prevention roles against bacterial infection,which not only helps us understand the structure and function of mapk genes in black rockfish,but also provides a reference to understand the role of mapk genes in teleost immune responses.展开更多
Long non-coding RNAs(lncRNAs)are a class of transcripts longer than 200 bp,which have been emerged as essential regulators in numerous biological processes.Black rockfish(Sebastes schlegelii)is an economic fish that w...Long non-coding RNAs(lncRNAs)are a class of transcripts longer than 200 bp,which have been emerged as essential regulators in numerous biological processes.Black rockfish(Sebastes schlegelii)is an economic fish that widely cultured in the coastal areas of China,Japan,and South Korea.With the expansion of aquacultural scale,various pathogens have threatened its industry and reduced its economic values.It has been reported that lncRNA were involved in the immune response and metabolic pathway in teleost,while no study is available on identification and functional analysis of lncRNAs in black rockfish so far.Herein,this study was performed to identify lncRNAs in the intestine of black rockfish after Edwardsiella tarda infection.In our results,a total of 9311 lncRNAs were identified through highthroughput sequencing,and 102 lncRNAs were significantly regulated following challenge,which were predicted to target 3348 mRNAs.Results of Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses of the se target genes showed they were function in catalytic activity,hydrolase activity,defense response and peptidase activity,which involved in metabolic pathways and immune related pathways.In addition,47 lncRNAs and 8 differentially expressed mRNAs(DEmRNAs)showed co-expression at two or more infection time points with metabolism and immunity functions.Moreover,real-time quantitative PCR(qRT-PCR)was performed to verify the reliability of sequencing gene expression analysis results.This research laid the foundation for further investigation of the regulatory roles of lncRNAs in the intestinal immune response of black rockfish.展开更多
CD209,a transmembrane lectin belonging to the C-type lectin family,can recognize carbohydrates on the surface of host cells and invading pathogens,and play an important role in cell adhesion and migration,pathogen rec...CD209,a transmembrane lectin belonging to the C-type lectin family,can recognize carbohydrates on the surface of host cells and invading pathogens,and play an important role in cell adhesion and migration,pathogen recognition and immune activation.Although well characterized in mammals,CD209 is still under-researched in fish.Here,we report a CD209-like gene,which was named SsCD209like,in black rockfish Sebastes schlegelii,and analyzed its structure features,expression patterns and ligand-binding activities.SsCD209like displays structural similarities to mammalian CD209s,with a cytosolic tail at N-terminus,a transmembrane region and an extracellular part containing a neck region and a CRD at C-terminus.The extracellular region and the neck region of SsCD-209like can both form dimers,which is different with the tetramer in human homologue.This result demonstrates the multimerization of CD209 homologue in fish for the first time.The EPN motif,a functional motif participating in sugar binding and affinity determination,is conserved in the CRD of SsCD209like,which is consistent with the higher binding strength of this lectin to L-fucose,D-GlcNAc and D-mannose.The binding of SsCD209like to different bacteria strains and bacteria-derived pathogen associated molecular patterns(PAMPs)are also observed in a dose-dependent manner.Results in this study show the sequence and structure features of SsCD209like and demonstrate its binding properties as a pathogen recognition receptor,which promotes our understanding of CD209 homologues in fish and provides basis for more in-depth studies of this molecule in the future.展开更多
Black rockfish Sebastes schlegelii juveniles （30-40 mm total length） were immersed in a range of calcein （CAL） solutions at concentrations ranging from 50 to 250 mg/L and alizarin red S （ARS） solutions at concen...Black rockfish Sebastes schlegelii juveniles （30-40 mm total length） were immersed in a range of calcein （CAL） solutions at concentrations ranging from 50 to 250 mg/L and alizarin red S （ARS） solutions at concentrations ranging from 100 to 500 mg/L in filtered seawater （salinity 30） for 24 h. Fluorescent marks were detected in otoliths （sagittae, asteriscus）, scales, fin rays （dorsal, pectoral, ventral, anal, and caudal fin rays）, and fin spines （dorsal, ventral, and anal fin spines） after a 60-d growth experiment. With the exception of 50-100 rng/L CAL, acceptable marks were produced in the otoliths and fin spines by all concentrations of CAL and ARS. In particular, marks were clearly visible under normal light in the sagittae, asteriscus, and fin spines offish immersed in 200 500 mg/L, 300-500 rag/L, and 200-500 mg/LARS, respectively. Scales and fin rays had acceptable marks at much higher concentrations （≥50 mg/L CAL, ≥300 mg/L ARS for scales and ≥50 mg/L CAL,≥200 mg/L ARS for fin rays）. The mark quality was highest （i.e., acceptable marks were observed in all sampled structures after immersion marking） in fish immersed in 150-250 mg/L CAL or 300-500 mg/LARS. In addition, there was no significant difference in survival and growth of marked fish compared with controls 60 d post-marking （P〉0.05）.展开更多
We tested the utility of chemical marking techniques in the juvenile black rockfish Sebastes schlegelii. Juveniles （30-40 mm total length） were immersed in a range of tetracycline hydrochloride （TC） solutions at c...We tested the utility of chemical marking techniques in the juvenile black rockfish Sebastes schlegelii. Juveniles （30-40 mm total length） were immersed in a range of tetracycline hydrochloride （TC） solutions at concentrations ranging from 300 to 500 mg/L, and alizarin complexone （ALC） solutions at concentrations ranging from 200 to 400 mg/L in filtered sea water （salinity of 30） for 24 h, respectively. Otoliths （sagittae, asteriscus）, scales, fin rays （dorsal, pectoral, ventral, anal, and caudal fin rays）, and fin spines （dorsal, ventral, and anal fin spines） were sampled and used to detect fluorescent marks after a 60-day growth experiment. With the exception of 300 mg/L TC, acceptable marks were produced in the otoliths and fin spines by all concentrations of TC and ALC. In particular, we observed clearly visible marks in the sagittae, asteriscus, and fin spines under normal light at concentrations of200~00 mg/L, 250-400 mg/L, and 250-400 mg/L ALC, respectively. Scales and fin rays had acceptable marks at much higher concentrations （_〉350 mg/L TC, 〉250 mg/L ALC for scales and _〉350 mg/L TC, 〉300 mg/L ALC for fin rays）. The best mark quality （i.e., acceptable marks were observed in all sampled structures after immersion marking） were obtained following immersion in TC at between 350-500 rag/L, and ALC between 300-400 mg/L. In addition, there was no significant difference in survival and growth of TC and ALC marked fish compared to their controls up to 60 days post-marking （P〉0.05）.展开更多
The structure of intestinal microbiota of black rockfish Sebastes schlegelii in five early development stages were determined in high throughput sequencing with Illumina MiSeq PE300 system.The relationship between int...The structure of intestinal microbiota of black rockfish Sebastes schlegelii in five early development stages were determined in high throughput sequencing with Illumina MiSeq PE300 system.The relationship between intestinal microbial community and the environmental(including culture water and feed)microbiota and the abundance variation trends of core microbiota were investigated,based on which the source of some core microbiota was analyzed in this study.The results show that Proteobacteria and Firmicutes are the most dominant phyla in guts.At the genus level,there are obvious differences between the artificial breeding fish and wild adults in the intestinal microflora structure.The compositions of dominant genera are similar,although the structure of intestinal microbiota gradually changes with the growth of larvae and juveniles.The core microbiota including Bacillus,Acinetobacter,Pseudomonas,Lactobacillus,Lactococcus,Glaciecola,Vibrio,Pseudoalteromonas,Acidovorax,and Aliivibrio were determined in the analysis of dominant and shared species.Compared with the water,the effect of feed microbiota on the structure of the gut microbial community is more obvious.Moreover,the trends of Bacillus,Acinetobacter,Pseudomonas,Lactobacillus,Lactococcus,and Glaciecola were opposite to Vibrio and Pseudoalteromonas in the gut.The correlation analysis suggested that Acidovorax,Glaciecola,Pseudomonas,Lactobacillus,and Acinetobacter might transited from mainly the parents and/or came from the fertilization process.The relative results may provide a theoretical reference for selecting the native probiotics,and supply the basic data for artificially regulating the intestinal microbiota with probiotic during early developmental stage of black rockfish.展开更多
Evolutionary evidence suggests that Sox3, a member of the high-mobility-group(HMG) family of transcription factors, is an ancestral precursor of Sry and is involved in sex determination similar to Sry. However, there ...Evolutionary evidence suggests that Sox3, a member of the high-mobility-group(HMG) family of transcription factors, is an ancestral precursor of Sry and is involved in sex determination similar to Sry. However, there is limited information regarding the SOX3 gene of the black rockfish(Sebastes schlegeli). In this study, we first isolated SOX3 gene from the gonads of S. schlegeli by homology cloning. The full-length of S. schlegeli SOX3(SsSOX3) c DNA was 1386 bp, comprising a 906-bp open reading frame, which encodes a peptide showing 93.6% and 93.9% homology with the Sox3 proteins of Epinephelus coioides and Oryzias latipe, respectively. Comparison of the cDNA sequence of the Ss SOX3 gene with the corresponding genomic DNA fragment revealed that the SsSOX3 gene consists of a single exon. Phylogenetic analysis demonstrated the evolutionary relationship of Ss SOX3 with other known SOXB1 genes in fish and tetrapods. The promoter region contains binding sites of several transcriptional factors that might participate in the regulation of Ss SOX3 expression. Quantitative real-time PCR analysis indicated that SsSOX3 was expressed in all the investigated larval developmental stages from 1 to 35 days after birth and the level of expression gradually decreased as the development proceeded. SsSOX3 exhibited sexually dimorphic expression in adult gonads, with high expression in the ovary but low expression in the testis. In situ hybridization revealed that SsSOX3 was strongly expressed in oocytes and follicular cells of ovaries but slightly expressed in germ cells of testicular tissues. Therefore, this study suggests that Ss SOX3 plays an important role in oogenesis and ovary differentiation in S. schlegeli.展开更多
Sex related FTZ-F1 is a transcriptional factor regulating the expression of fushi tarazu (a member of the orphan nuclear receptors) gene. In this study, FTZ-F1 gene (FTZ-F1) was isolated from the testis of black rockf...Sex related FTZ-F1 is a transcriptional factor regulating the expression of fushi tarazu (a member of the orphan nuclear receptors) gene. In this study, FTZ-F1 gene (FTZ-F1) was isolated from the testis of black rockfish (Sebastes schlegeli) by homology cloning. The full-length cDNA of S. schlegeli FTZ-F1 (ssFTZ-F1) contained a 232bp 5′UTR, a 1449bp ORF encoding FTZ-F1 (482 amino acid residules in length) with an estimated molecular weight of 5.4kD and a 105bp 3′UTR. Sequence, tissue distribution and phylogenic analysis showed that ssFTZ-F1 belonged to FTZ group, holding highly conserved regions including Ⅰ, Ⅱ and Ⅲ FTZ-F1 boxes and an AF-2 hexamer. Relatively high expression was observed at different larva stages. In juveniles (105 days old), the transcript of ssFTZ-F1 can be detected in all tissues and the abuncance of the gene transcript in testis, ovary, spleen and brain was higher than that in other tissues. In mature fish, the abundance of gene transcript was higher in testis, ovary, spleen and brain than that in liver (trace amount), and the gene was not transcribed in other tissues. The highest abundance of gene transcript was always observed in gonads of both juvenile and mature fish. In addition, the abundance of gene transcript in male tissues were higher than that in female tissue counterparts (P<0.05).展开更多
The assessment of population structure and abundance of fish assemblages associated with artificial reefs(ARs) is an important aspect of AR management.In the present study,we used a DiveOperated Stereo Video(stereo-DO...The assessment of population structure and abundance of fish assemblages associated with artificial reefs(ARs) is an important aspect of AR management.In the present study,we used a DiveOperated Stereo Video(stereo-DOV) technique to assess the population structure and abundance of Sebastes schlegeli associated with two metallic,and three wooden,vessel reefs in Haizhou Bay during 2012 and2013.The study used video systems to obtain length measurements and estimates of abundance.The size composition of S.schlegeli differed among reefs and individuals around vessel reefs were all adults,with total lengths(TL) of >20 cm.Juvenile fish were encountered by divers in a rocky area near the island away from the vessel reefs.The largest individual S.schlegeli(with the highest TL) among five reefs were found around a metallic vessel reef in both 2012 and 2013.TL of S.schlegeli from all reefs increased by an average of 3.2 cm(P<0.05) from 2012 to 2013,with an estimated mean weight increase of 250.4 g(P<0.05).The video survey also indicated a decrease in the biomass of schools near two metallic vessels between the years.Stereo-video technology was found to be suitable for rockfish surveys around the reefs.展开更多
Ovoviviparous black rockfish(Sebastes schlegeli) is an important marine fish species for aquaculture and fisheries in China. Genetic information of this species is scarce because of the lack of microsatellite markers....Ovoviviparous black rockfish(Sebastes schlegeli) is an important marine fish species for aquaculture and fisheries in China. Genetic information of this species is scarce because of the lack of microsatellite markers. In this study, a large number of microsatellite markers of black rockfish were isolated by constructing microsatellite-enriched libraries. Female- and male-specific genetic linkage maps were constructed using 435 microsatellite markers genotyped in a full-sib family of the fish species. The female linkage map contained 140 microsatellite markers, in which 23 linkage groups had a total genetic length of 1334.1 c M and average inter-marker space of 13.3 c M. The male linkage map contained 156 microsatellite markers, in which 25 linkage groups had a total genetic length of 1359.6 c M and average inter-marker distance of 12.4 c M. The genome coverage of the female and male linkage maps was 68.6% and 69.3%, respectively. The female-to-male ratio of the recombination rate was approximately 1.07:1 in adjacent microsatellite markers. This paper presents the first genetic linkage map of microsatellites in black rockfish. The collection of polymorphic markers and sex-specific linkage maps of black rockfish could be useful for further investigations on parental assignment, population genetics, quantitative trait loci mapping, and marker-assisted selection in related breeding programs.展开更多
基金supported by the National Natural Science Foundation of China(Nos.32273133,31970492).
文摘Black rockfish(Sebastes schlegelii)has evolved viviparous reproduction mode.Different from the oviparous fishes,the spermatozoa of black rockfish are stored on the surface of ovarian follicle for over four months until the egg mature in ovary,while the adaptive evolution of fertilization-related genes remains to be studied.In the present study,based on the genome and transcriptome information of black rockfish,a total of 10 hyaluronidase genes were identified by phylogenetic and sequence analyses,including hyal1,hyal2,hyal3,hyal4,hyal6 and spam1a,spam1b,spam1c and spam1d.The spam1 subfamilies,including spam1a,spam1b,spam1c and spam1d,were significantly expanded compared to other species.The adaptive evolution of hyaluronidase was further investigated by selection pressure analysis of branch model and branch site model.The results showed that only spam1 subfamily was positively selected with a large number of positive selection sites,and the evolution rate was significantly higher thanthose of other teleosts.Two positively selected sites,LYS-171 and GLY-164,were located in the neutral hyaluronidase activity domain,which was pivotal for SPAM1 to participate in enzymatic hydrolysis of the extracellular hyaluronic acid matrix and mediate acrosome reaction in mammals.Furthermore,spam1 was mainly expressed in spermatocytes,secondary spermatocytes,and spermatids of testis,whereas it was not found in metamorphic sperm and mature sperm based on the spatiotemporal expression analysis.All results indicated that spam1 might originate from the testis and might be an adaptation of viviparous trait in black rockfish.
基金Supported by the National Key R&D Program of China(No.2018YFD0900101)the Young Experts of Taishan Scholars(No.tsqn201909130)+2 种基金the Science and Technology Support Plan for Youth Innovation of Colleges and Universities in Shandong Province(No.2019KJF003)the“First Class Fishery Discipline”Program in Shandong Provincethe Shandong Technical System of Fish Industry(No.SDAIT-12-03)。
文摘Mitogen-activated protein kinases(MAPKs)play pivotal roles in response to environmental stresses and bacterial infections.Compared with those in the higher vertebrates,studies of mapk gene family are still limited in teleost.Identification,characterization,classification,and expression profiling of totally 15 mapk genes in black rockfish(Sebastes schlegelii)were conducted.Phylogenetic relationships show that these mapk genes could be divided into extracellular signal-regulated kinase(ERK),c-Jun N-terminal kinase(JNK),and p38 sub-families.In addition,gene structures,syntenic analysis,and selective pressure analysis are performed to confirm their annotations.Results of selective pressure analysis indicate that mapk1,mapk3,mapk7,mapk10,mapk11,and mapk12 underwent significantly-positive selections,while the others genes such as mapk4,mapk6,mapk15,mapk8a,mapk8b,mapk9,mapk13,mapk14a,and mapk14b were under purifying selections.Moreover,results of qRT-PCR indicate that mapk genes in 8 healthy tissues displayed different expression patterns.The expression patterns of several mapk genes including mapk12,mapk13,mapk14a,mapk14b,and mapk15 were significantly changed in mucosal tissues after Edwardsiella piscicida infection.This study demonstrates that mapk genes in black rockfish play vital prevention roles against bacterial infection,which not only helps us understand the structure and function of mapk genes in black rockfish,but also provides a reference to understand the role of mapk genes in teleost immune responses.
基金Supported by the Young Experts of Taishan Scholars(No.tsqn201909130)the Science and Technology Support Plan for Youth Innovation of Colleges and Universities in Shandong Province(No.2019KJF003)+1 种基金the“First Class Fishery Discipline”Program in Shandong Province,a special talent program“One Matter One Decision(Yi Shi Yi Yi)”Program in Shandong Province,Chinathe Breeding Plan of Shandong Provincial Qingchuang Research Team(2019)。
文摘Long non-coding RNAs(lncRNAs)are a class of transcripts longer than 200 bp,which have been emerged as essential regulators in numerous biological processes.Black rockfish(Sebastes schlegelii)is an economic fish that widely cultured in the coastal areas of China,Japan,and South Korea.With the expansion of aquacultural scale,various pathogens have threatened its industry and reduced its economic values.It has been reported that lncRNA were involved in the immune response and metabolic pathway in teleost,while no study is available on identification and functional analysis of lncRNAs in black rockfish so far.Herein,this study was performed to identify lncRNAs in the intestine of black rockfish after Edwardsiella tarda infection.In our results,a total of 9311 lncRNAs were identified through highthroughput sequencing,and 102 lncRNAs were significantly regulated following challenge,which were predicted to target 3348 mRNAs.Results of Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses of the se target genes showed they were function in catalytic activity,hydrolase activity,defense response and peptidase activity,which involved in metabolic pathways and immune related pathways.In addition,47 lncRNAs and 8 differentially expressed mRNAs(DEmRNAs)showed co-expression at two or more infection time points with metabolism and immunity functions.Moreover,real-time quantitative PCR(qRT-PCR)was performed to verify the reliability of sequencing gene expression analysis results.This research laid the foundation for further investigation of the regulatory roles of lncRNAs in the intestinal immune response of black rockfish.
基金financially supported by the National Natural Science Foundation of China (No.32002422)the Natural Science Foundation of Shandong Province (No.ZR 2020QC212)+4 种基金the Young Experts of Taishan Scholars (No.tsqn201909130)the Shandong Technical System of Fish Industry (No. SDAIT-12-03)the Science and Technology Support Plan for Youth Innovation of Colleges and Universities in Shandong Province (No. 2019KJF003)the Breeding Plan of Shandong Provincial Qingchuang Research Team (2019)the Advanced Talents Foundation of QAU Grant (No. 663-1120023)
文摘CD209,a transmembrane lectin belonging to the C-type lectin family,can recognize carbohydrates on the surface of host cells and invading pathogens,and play an important role in cell adhesion and migration,pathogen recognition and immune activation.Although well characterized in mammals,CD209 is still under-researched in fish.Here,we report a CD209-like gene,which was named SsCD209like,in black rockfish Sebastes schlegelii,and analyzed its structure features,expression patterns and ligand-binding activities.SsCD209like displays structural similarities to mammalian CD209s,with a cytosolic tail at N-terminus,a transmembrane region and an extracellular part containing a neck region and a CRD at C-terminus.The extracellular region and the neck region of SsCD-209like can both form dimers,which is different with the tetramer in human homologue.This result demonstrates the multimerization of CD209 homologue in fish for the first time.The EPN motif,a functional motif participating in sugar binding and affinity determination,is conserved in the CRD of SsCD209like,which is consistent with the higher binding strength of this lectin to L-fucose,D-GlcNAc and D-mannose.The binding of SsCD209like to different bacteria strains and bacteria-derived pathogen associated molecular patterns(PAMPs)are also observed in a dose-dependent manner.Results in this study show the sequence and structure features of SsCD209like and demonstrate its binding properties as a pathogen recognition receptor,which promotes our understanding of CD209 homologues in fish and provides basis for more in-depth studies of this molecule in the future.
基金Supported by the Special Fund for Agro-scientific Research in the Public Interest(No.201003068)the National Natural Science Foundation of China(Nos.31172447,41176117)
文摘Black rockfish Sebastes schlegelii juveniles （30-40 mm total length） were immersed in a range of calcein （CAL） solutions at concentrations ranging from 50 to 250 mg/L and alizarin red S （ARS） solutions at concentrations ranging from 100 to 500 mg/L in filtered seawater （salinity 30） for 24 h. Fluorescent marks were detected in otoliths （sagittae, asteriscus）, scales, fin rays （dorsal, pectoral, ventral, anal, and caudal fin rays）, and fin spines （dorsal, ventral, and anal fin spines） after a 60-d growth experiment. With the exception of 50-100 rng/L CAL, acceptable marks were produced in the otoliths and fin spines by all concentrations of CAL and ARS. In particular, marks were clearly visible under normal light in the sagittae, asteriscus, and fin spines offish immersed in 200 500 mg/L, 300-500 rag/L, and 200-500 mg/LARS, respectively. Scales and fin rays had acceptable marks at much higher concentrations （≥50 mg/L CAL, ≥300 mg/L ARS for scales and ≥50 mg/L CAL,≥200 mg/L ARS for fin rays）. The mark quality was highest （i.e., acceptable marks were observed in all sampled structures after immersion marking） in fish immersed in 150-250 mg/L CAL or 300-500 mg/LARS. In addition, there was no significant difference in survival and growth of marked fish compared with controls 60 d post-marking （P〉0.05）.
基金Supported by the Special Fund for Agro-Scientific Research in the Public Interest(No.201003068)the National Natural Science Foundation of China(Nos.31172447,41176117)
文摘We tested the utility of chemical marking techniques in the juvenile black rockfish Sebastes schlegelii. Juveniles （30-40 mm total length） were immersed in a range of tetracycline hydrochloride （TC） solutions at concentrations ranging from 300 to 500 mg/L, and alizarin complexone （ALC） solutions at concentrations ranging from 200 to 400 mg/L in filtered sea water （salinity of 30） for 24 h, respectively. Otoliths （sagittae, asteriscus）, scales, fin rays （dorsal, pectoral, ventral, anal, and caudal fin rays）, and fin spines （dorsal, ventral, and anal fin spines） were sampled and used to detect fluorescent marks after a 60-day growth experiment. With the exception of 300 mg/L TC, acceptable marks were produced in the otoliths and fin spines by all concentrations of TC and ALC. In particular, we observed clearly visible marks in the sagittae, asteriscus, and fin spines under normal light at concentrations of200~00 mg/L, 250-400 mg/L, and 250-400 mg/L ALC, respectively. Scales and fin rays had acceptable marks at much higher concentrations （_〉350 mg/L TC, 〉250 mg/L ALC for scales and _〉350 mg/L TC, 〉300 mg/L ALC for fin rays）. The best mark quality （i.e., acceptable marks were observed in all sampled structures after immersion marking） were obtained following immersion in TC at between 350-500 rag/L, and ALC between 300-400 mg/L. In addition, there was no significant difference in survival and growth of TC and ALC marked fish compared to their controls up to 60 days post-marking （P〉0.05）.
基金Supported by the Central Public-interest Scientific Institution Basal Research Fundthe CAFS+4 种基金the Fund of Key Laboratory of Sustainable Development of Marine Fisheries,Ministry of Agriculture,China(No.2018HY-XKQ01-04)the Marine S&T Fund of Shandong Province for Pilot National Laboratory for Marine Science and Technology(Qingdao)(No.2018SDKJ0501-2)the Central Public-interest Scientific Institution Basal Research Fund,YSFRI,CAFS,China(No.20603022018021)the China Agriculture Research System(No.CARS-47)the Key Project of Research and Development Plan of China(No.2017YFE0104400)
文摘The structure of intestinal microbiota of black rockfish Sebastes schlegelii in five early development stages were determined in high throughput sequencing with Illumina MiSeq PE300 system.The relationship between intestinal microbial community and the environmental(including culture water and feed)microbiota and the abundance variation trends of core microbiota were investigated,based on which the source of some core microbiota was analyzed in this study.The results show that Proteobacteria and Firmicutes are the most dominant phyla in guts.At the genus level,there are obvious differences between the artificial breeding fish and wild adults in the intestinal microflora structure.The compositions of dominant genera are similar,although the structure of intestinal microbiota gradually changes with the growth of larvae and juveniles.The core microbiota including Bacillus,Acinetobacter,Pseudomonas,Lactobacillus,Lactococcus,Glaciecola,Vibrio,Pseudoalteromonas,Acidovorax,and Aliivibrio were determined in the analysis of dominant and shared species.Compared with the water,the effect of feed microbiota on the structure of the gut microbial community is more obvious.Moreover,the trends of Bacillus,Acinetobacter,Pseudomonas,Lactobacillus,Lactococcus,and Glaciecola were opposite to Vibrio and Pseudoalteromonas in the gut.The correlation analysis suggested that Acidovorax,Glaciecola,Pseudomonas,Lactobacillus,and Acinetobacter might transited from mainly the parents and/or came from the fertilization process.The relative results may provide a theoretical reference for selecting the native probiotics,and supply the basic data for artificially regulating the intestinal microbiota with probiotic during early developmental stage of black rockfish.
基金supported by the National Natural Science Foundation of China (No. 31372511)
文摘Evolutionary evidence suggests that Sox3, a member of the high-mobility-group(HMG) family of transcription factors, is an ancestral precursor of Sry and is involved in sex determination similar to Sry. However, there is limited information regarding the SOX3 gene of the black rockfish(Sebastes schlegeli). In this study, we first isolated SOX3 gene from the gonads of S. schlegeli by homology cloning. The full-length of S. schlegeli SOX3(SsSOX3) c DNA was 1386 bp, comprising a 906-bp open reading frame, which encodes a peptide showing 93.6% and 93.9% homology with the Sox3 proteins of Epinephelus coioides and Oryzias latipe, respectively. Comparison of the cDNA sequence of the Ss SOX3 gene with the corresponding genomic DNA fragment revealed that the SsSOX3 gene consists of a single exon. Phylogenetic analysis demonstrated the evolutionary relationship of Ss SOX3 with other known SOXB1 genes in fish and tetrapods. The promoter region contains binding sites of several transcriptional factors that might participate in the regulation of Ss SOX3 expression. Quantitative real-time PCR analysis indicated that SsSOX3 was expressed in all the investigated larval developmental stages from 1 to 35 days after birth and the level of expression gradually decreased as the development proceeded. SsSOX3 exhibited sexually dimorphic expression in adult gonads, with high expression in the ovary but low expression in the testis. In situ hybridization revealed that SsSOX3 was strongly expressed in oocytes and follicular cells of ovaries but slightly expressed in germ cells of testicular tissues. Therefore, this study suggests that Ss SOX3 plays an important role in oogenesis and ovary differentiation in S. schlegeli.
基金supported by the National High-Tech Research and Development Program (2012AA10A402)the National Natural Science Foundation of China(31172385)
文摘Sex related FTZ-F1 is a transcriptional factor regulating the expression of fushi tarazu (a member of the orphan nuclear receptors) gene. In this study, FTZ-F1 gene (FTZ-F1) was isolated from the testis of black rockfish (Sebastes schlegeli) by homology cloning. The full-length cDNA of S. schlegeli FTZ-F1 (ssFTZ-F1) contained a 232bp 5′UTR, a 1449bp ORF encoding FTZ-F1 (482 amino acid residules in length) with an estimated molecular weight of 5.4kD and a 105bp 3′UTR. Sequence, tissue distribution and phylogenic analysis showed that ssFTZ-F1 belonged to FTZ group, holding highly conserved regions including Ⅰ, Ⅱ and Ⅲ FTZ-F1 boxes and an AF-2 hexamer. Relatively high expression was observed at different larva stages. In juveniles (105 days old), the transcript of ssFTZ-F1 can be detected in all tissues and the abuncance of the gene transcript in testis, ovary, spleen and brain was higher than that in other tissues. In mature fish, the abundance of gene transcript was higher in testis, ovary, spleen and brain than that in liver (trace amount), and the gene was not transcribed in other tissues. The highest abundance of gene transcript was always observed in gonads of both juvenile and mature fish. In addition, the abundance of gene transcript in male tissues were higher than that in female tissue counterparts (P<0.05).
基金Supported by the National Natural Science Foundation of China(NSFC)-Shandong Joint Fund for Marine Science Research Centers(No.U1406403)the National Key Technology Research and Development Program of China(No.2011BAD13B02)+1 种基金the National Marine Public Welfare Research Project(No.201205023)the Key Laboratory of Marine Ecology and Environmental Science and Engineering,State Oceanic Administration(No.MESE-2013-05)
文摘The assessment of population structure and abundance of fish assemblages associated with artificial reefs(ARs) is an important aspect of AR management.In the present study,we used a DiveOperated Stereo Video(stereo-DOV) technique to assess the population structure and abundance of Sebastes schlegeli associated with two metallic,and three wooden,vessel reefs in Haizhou Bay during 2012 and2013.The study used video systems to obtain length measurements and estimates of abundance.The size composition of S.schlegeli differed among reefs and individuals around vessel reefs were all adults,with total lengths(TL) of >20 cm.Juvenile fish were encountered by divers in a rocky area near the island away from the vessel reefs.The largest individual S.schlegeli(with the highest TL) among five reefs were found around a metallic vessel reef in both 2012 and 2013.TL of S.schlegeli from all reefs increased by an average of 3.2 cm(P<0.05) from 2012 to 2013,with an estimated mean weight increase of 250.4 g(P<0.05).The video survey also indicated a decrease in the biomass of schools near two metallic vessels between the years.Stereo-video technology was found to be suitable for rockfish surveys around the reefs.
基金supported by the National High-Tech Research and Development Program (Grant Nos.2012AA10A408 and 2012AA10A402)
文摘Ovoviviparous black rockfish(Sebastes schlegeli) is an important marine fish species for aquaculture and fisheries in China. Genetic information of this species is scarce because of the lack of microsatellite markers. In this study, a large number of microsatellite markers of black rockfish were isolated by constructing microsatellite-enriched libraries. Female- and male-specific genetic linkage maps were constructed using 435 microsatellite markers genotyped in a full-sib family of the fish species. The female linkage map contained 140 microsatellite markers, in which 23 linkage groups had a total genetic length of 1334.1 c M and average inter-marker space of 13.3 c M. The male linkage map contained 156 microsatellite markers, in which 25 linkage groups had a total genetic length of 1359.6 c M and average inter-marker distance of 12.4 c M. The genome coverage of the female and male linkage maps was 68.6% and 69.3%, respectively. The female-to-male ratio of the recombination rate was approximately 1.07:1 in adjacent microsatellite markers. This paper presents the first genetic linkage map of microsatellites in black rockfish. The collection of polymorphic markers and sex-specific linkage maps of black rockfish could be useful for further investigations on parental assignment, population genetics, quantitative trait loci mapping, and marker-assisted selection in related breeding programs.
