Molecular Characterization and Antibacterial Activity of a G-Type Lysozyme in Yellow Drum(Nibea albiflora)

Lysozyme(EC3.2.1.17)plays an important role in the immune response;as a nonspecific immune factor,it can resist causative agents.Lysozyme can be divided into c-type and g-type in fish.In a previous study,through genome-wide association analysis,the g-type lysozyme gene,which is named NaLyg in yellow drum(Nibea albiflora),was found to be a key candidate gene for disease resistance in response to Vibrio harveyi infection.The cDNA of NaLyg was 1025 bp,including four exons and three introns,and its open reading frame(ORF)had a full-length of 582 bp,encoding 193 amino acids.NaLyg was found to be conserved during evolution through bioinformatic analyses.The NaLyg protein possessed a sugar binding domain and three catalytic sites,including Glu71,Asp84 and Asp101.Quantitative qRT-PCR results confirmed that NaLyg gene mRNA was visibly increased after V.harveyi infection.The NaLyg protein purified by prokaryotic expression killed some gram-negative bacterial pathogens by inducing cell wall destruction,including V.harveyi,Aeromonas hydrophila and Edwardsiella tarda.Moreover,the NaLyg protein killed two gram-positive bacteria,Bacillus subtilis and Staphylococcus aureus.Taken together,the experimental results suggested that the NaLyg protein of N.albiflora played an important role in fighting bacterial infections.

Identification,Characterization,and Probiotic Potentials of Lactobacillus pentosus SF-1

In recent years probiotics have been considered as a potential substitution of antibiotics to control pathogens and treat infectious diseases in aquaculture.In the present study a strain of Lactobacillus pentosus,named as L.pentosus SF-1,was isolated from waters in aquaculture.The species identification of this strain was conducted by 16S rRNA sequence,and the physiological and biochemical characteristics of this strain were assessed.Furthermore,the virulence,antibiotic sensitivity,cell surface characteristics and acid/base-resistance of L.pentosus SF-1 were determined to evaluate the probiotic potentials of this strain.Specifically,L.pentosus SF-1 is sensitive to most common antibiotics,and no hemolysin was generated from it,indicating the safety of this strain to hosts.In addition,L.pentosus SF-1 was able to tolerate the artificial gastric juice at pH 3 for 4 h and the artificial intestinal fluid at pH 6.8 or 8.0 for 6 h.Moreover,the analysis of self-aggregation and the adhesion of L.pentosus SF-1 to organic solvents suggested a high potential of L.pentosus SF-1 to inhabit the hosts,which was confirmed by testing the colonization of L.pentosus SF-1 in germ-free zebrafish.Interestingly,L.pentosus SF-1 displayed a high bactericidal activity against several bacterial pathogens.Consistently,the incubation of L.pentosus SF-1 significantly promoted the expression of antimicrobial components in zebrafish,contributing to the protection of the fish from E.tarda infection in vivo.Taken together,the probiotic strain L.pentosus SF-1 could be applied as anti-infection reagent in aquaculture.

Transcriptome Analysis of Heterosis in Survival in the Hybrid Progenies of‘Haida No.1’and Orange-Shelled Lines of the Pacific Oyster Crassostrea gigas

Heterosis has been exploited to enhance the yield and adaptability in various shellfish species;however,the molecular basis of it remains unclear.The Pacific oyster Crassostrea gigas is one of the most economically important aquaculture species,and its productive traits can be improved by hybridization.Here,an intraspecific cross between orange shell(O,10th generation)and‘Haida No.1’(H,13th generation)of C.gigas was performed to assess the heterosis of survival trait.Survival rates of hybrid family(OH)and inbred families(HH and OO)were compared at larval stage,and eyed-pediveliger larvae of three families were subjected to transcriptome analysis.The analysis results of best-parent heterosis and mid-parent heterosis showed that the hybrid family exhi-bited a high heterosis in survival relative to the parental families.The OH-M(OH vs.OO)and OH-P(OH vs.HH)had 425 and 512 dif-ferentially expressed genes(DEGs),respectively.Functional enrichment analysis of these DEGs revealed that the significantly enrich-ed genes function in virion binding,C-type lectin receptor signaling pathway,cellular defense response and other immune-related pro-cesses,which involves perlucin-like protein,CD209 antigen-like protein,ZNFX1,caspase-3 and acan genes.These differentially ex-pressed genes in OH-M and OH-P,together with the immune-related processes mentioned above may play an important role in the larval survival of C.gigas.In addition,three genes(CYP450,fucolectin and perlucin-like)are associated with the orange shell and low survival of maternal oyster OO.These findings provide support for the application of hybrid with superior survival and will facilitate the understanding of heterosis formation in the Pacific oyster.

Assessment of the Characteristics of Demersal Fish Communities Using Species-and Trait-Based Approaches in the Coastal Habitats Along Rongcheng Bay,Shandong Peninsula,China

Biodiversity declines have motivated many studies on the relationship between species diversity and ecosystem functioning.In this study,we described the spatial-temporal characteristics of demersal fish communities along a coastal habitat in Rongcheng Bay,Shandong Peninsula,China with both species-based and biological trait-based approaches.The field survey was carried out monthly using traps from April to October of 2018,and divided into three seasons(spring:April and May;summer:June,July and August;autumn:September,October and November).The study area included five distinct habitats:seagrass bed,natural rocky reef,bare sand,artificial reef together with natural rocky reef,and artificial reef together with bare sand.We analyzed the fish communities with three taxonomic diversity indices,including Shannon-Wiener,Simpson,and Pielou Evenness,as well as four functional diversity indices,including FRic,FEve,FDiv,and FDis,based on 7 functional groups which are categorized into 27 traits.The results showed that there were no significant differences in taxonomic diversity indices among different habitats in the three seasons.However,significant differences were found in the functional richness of fish communities among different habitats in three seasons.Seagrass bed represented the highest functional richness in spring and autumn.This study demonstrates that seagrass bed is very important in enhancing the functional diversity of fish communities in a complex habitat.The study also indicates that the combination of taxonomic diversity and functional diversity will provide a more detailed description of the characteristics of fish communities.

Dietary Supplementation of β-Carotene Reveals miRNAs Involved in the Regulation of Carotenoid Metabolism in Crassostrea gigas

Carotenoids play crucial physiological roles in animals.A comprehensive investigation into the mechanism of carotenoid metabolism in oysters will establish a theoretical foundation for further development of its carotenoid-rich traits.However,the information on the function of miRNA in β-carotene metabolism in oysters is limited.To elucidate the mechanisms underlying miRNA regulation of carotenoid metabolism in oysters,we compared the expressions of miRNA in digestive gland tissues of Pacific oyster(Crassostrea gigas)fed with aβ-carotene supplemented diet and a normal diet,respectively.A total of 690 candidate miRNAs in the Pacific oyster digestive gland tissues were identified,including 590 known miRNAs and 111 unknown miRNAs.Three differentially expressed miRNAs were obtained in the carotenoid-fed and normal groups,associated to 137 differentially expressed target genes.Moreover,the GO enrichment analysis revealed that the differentially expressed target genes were mainly involved in transmembrane transport activity.KEGG enrichment showed that the differentially expressed target genes were involved in ABC transport.Analysis of the mRNA-miRNA network revealed that novel0025 played a central role in carotenoid metabolism,and it was negatively correlated with the expression of 46 mRNAs.In addition,down-regulated expression of novel0025 upregulated the expression of the lipoprotein gene LOC105342186,suggesting a potential regulatory role in carotenoid metabolism.Our results provide useful information for elucidating the miRNA regulation mechanism during carotenoids metabolism in the Pacific oyster.

Non-Targeted Metabolomics Reveals the Metabolic Alterations in Response to Artificial Selective Breeding in the Fast-Growing Strains of Pacific Oyster

Pacific oyster(Crassostrea gigas)is one of the most important mollusks cultured all around the world.Selective breeding programs of Pacific oysters in China is initiated since 2006 and developed the genetically improved strain with fast-growing trait.However,little is known about the metabolic signatures of the fast-growing trait.In the present study,the non-targeted metabolomics was performed to analyze the metabolic signatures of adductor muscle tissue in one-year old Pacific oysters from fast-growing strain and the wild population.A total of 7767 and 10174 valid peaks were extracted and quantified in ESI^(+)and ESI^(−)modes,resulting in 399 and 381 annotated metabolites,respectively.PCA and OPLS-DA revealed that considerable separation among samples from fastgrowing strain and wild population,suggesting the differences in metabolic signatures.Meanwhile,81 significantly different metabolites(SDMs)were identified in the comparisons between fast-growing strain and wild population,based on the strict thresholds.It was found that there were highly correlation and conserved coordination among these SDMs.KEGG enrichment analysis indicated that the SDMs were tightly related to pantothenate and CoA biosynthesis,steroid hormone biosynthesis,riboflavin metabolism,and arginine and proline metabolism.Of them,the CoA biosynthesis and metabolism,affected by pantetheine and pantothenic acid,might be important for the growth of Pacific oysters under artificial selective breeding.The study provides the comprehensive views of metabolic signatures in response to artificially selective breeding,and is helpful to better understand the molecular mechanism of fastgrowing traits in Pacific oysters.

An Improved Chromosome-Level Genome Assembly and Annotation of Belted Beard Grunt(Hapalogenys analis)

Hapalogenys analis(order Lobotiformes)is an economically and ecologically significant fish species.It is a typical sedentary rocky reef fish and is primarily found in the northern Pacific Ocean.Here,we used Hi-C and PacBio sequencing technique to assemble a high-quality,chromosome-level genome for this species.The 539 Mb genome had a contig N50 with a size of 3.43 Mb,while 755 contigs clustered into 24 chromosomal groups with an anchoring rate of 99.02%.Of the total genomic sequence,132.74Mb(24.39%)were annotated as repeat elements.A total of 21360 protein-coding genes were identified,of which 20787 genes(97.32%)were successfully annotated to public databases.The BUSCO evaluation indicated that 96.90%of the total orthologous genes were matched.The phylogenetic tree representing H.analis and 14 other bony fish species indicated that the H.analis genome contained 364 expanded gene families related to olfactory receptor activity,compared with the common ancestor of H.analis and Sciaenidae.Comparative genomic analysis further identified 3584 contracted gene families.Branch-site modeling identified 277 genes experiencing positive selection,which may facilitate the adaptation to rocky reef environments.The genome reported here is helpful for ecological and evolutionary studies of H.analis.

Molecular phylogenetic relationships based on mitochondrial genomes of novel deep-sea corals(Octocorallia:Alcyonacea):Insights into slow evolution and adaptation to extreme deep-sea environments

A total of 10 specimens of Alcyonacea corals were collected at depths ranging from 905 m to 1633 m by the manned submersible Shenhai Yongshi during two cruises in the South China Sea(SCS).Based on mitochondrial genomic characteristics,morphological examination,and sclerite scanning electron microscopy,the samples were categorized into four suborders(Calcaxonia,Holaxonia,Scleraxonia,and Stolonifera),and identified as 9 possible new cold-water coral species.Assessments of GC-skew dissimilarity,phylogenetic distance,and average nucleotide identity(ANI)revealed a slow evolutionary rate for the octocoral mitochondrial sequences.The nonsynonymous(Ka)to synonymous(Ks)substitution ratio(Ka/Ks)suggested that the 14 protein-coding genes(PCGs)were under purifying selection,likely due to specific deep-sea environmental pressures.Correlation analysis of the median Ka/Ks values of five gene families and environmental factors indicated that the genes encoding cytochrome b(cyt b)and DNA mismatch repair protein(mutS)may be influenced by environmental factors in the context of deep-sea species formation.This study highlights the slow evolutionary pace and adaptive mechanisms of deep-sea corals.

Impacts of Integrated Multi-Trophic Aquaculture on Phytoplankton in Sanggou Bay

Integrated multi-trophic aquaculture(IMTA)has been considered as an ecofriendly culture system providing a potential solution to environmental risks caused by intensive monoculture system.However,the impact of IMTA on phytoplankton remains unclear.In this study,the spatial and temporal variations of phytoplankton in Sanggou Bay were investigated seasonally based on 21 sampling sites covering three cultivation zones(bivalve zone,IMTA zone,and kelp zone)and one control zone(without aquatic cultivation).In total,128 phytoplankton species,with diatoms and dinoflagellates as the dominant groups,were obtained across the whole year,and the mean Shannon diversity index(H')and species richness(SR)were determined as 1.39 and 9.39,respectively.The maximum chlorophyll a(Chl-a)(6.32μg L^(-1))and plankton diversity(H'of 1.97)occurred in summer and autumn,respectively.Compared to other zones,the bivalve zone displayed significantly higher Chl-a and lower H'in majority of time.Pairwise PERMANOVA analysis indicated that the phytoplankton assemblage in the bivalve zone was significantly different with the control and kelp zones,while the IMTA zone maintained close to other three zones.Based on generalized additive models,temperature,NO_(2)^(-)-N,N/P ratio,SiO_(3)^(2-)-Si,and salinity were determined as the key factors underlying Chl-a and phytoplankton diversity.Addi-tionally,the results of redundancy analysis further indicated that the phytoplankton assemblage in the bivalve zone is positively re-lated with nutrients such as NO_(3)^(-)-N and NH_(4)^(+)-N as well as water depth,while the phytoplankton assemblages in the kelp,control,and IMTA zones are associated with NO_(2)^(-)-N,SiO_(3)^(2-)-Si,and salinity.Taken all observations into consideration together,it can be inferred that IMTA can effectively reduce Chl-a level compared to bivalve monoculture by reducing the nutrients.However,the SR,H’,and species composition of phytoplankton are primarily determined by local environment factors such as temperature,water depth,salinity and SiO_(3)^(2-)-Si.

Pigment Distribution and Secretion in the Mantle of the Pacific Oyster(Crassostrea gigas)

The color of Mollusca shells is one of the most important attributes to consumers.At the cellular level,black color is mainly from the melanin produced by melanocytes.The melanosome is a specialized membrane-bound organelle that is involved in melanin synthesis,storage,and transportation.How the complex pigmentation process in the Crassostrea gigas is established remains an open question.The objectives of this studies are to examine the morphological characteristics of melanosomes or melanin of mantle pigmentation in the Pacific oyster,thereby investigating its contribution to shell color.The results show that pigmented granules of the mantles vary among the three lobes,and the melanosomes at different stages are enriched in distinct cargo molecules,which indicate the remarkable difference between the marginal mantle and central mantle.Examination of mantle histology reveals that the mantle margin of the oyster is characterized by three different folds,including the outer secretory,middle sensory,and inner muscular fold.Ferrous ion chelating assays against the tyrosine hydroxylase indicate that a large amount of melanin is localized in the inner surface of the middle fold.Transmission electron microscopy analyses show that the mantle edge is composed of tall columnar and cuboidal epidermal cells and some pigmented melanocytes intersperse among these cells.The numbers of melanosomes among the three lobes are different.In the inner fold and the middle fold of the mantle,some single dispersion,or aggregation of melanosomes with different degrees of melanization are found in the outer surface.Numerous melanosomes are distributed in the epithelium of the outer fold of the mantle,and mainly are at the apical microvillar surface near the lumen.However,melanosomes are occasionally observed in the central mantle,and they are relatively less.This work provides new insights into the process of melanin deposit in the mantle and shell pigmentation in C.gigas.

Genome-wide characterization of mapk gene family in black rockfish Sebastes schlegelii and their expression patterns against Edwardsiella piscicida infection

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.

Early life history traits of chub mackerel Scomber japonicus in the Oyashio water revealed by otolith microstructure

Information on survival and growth during the early life stage is essential to understand the mechanism of interannual variations in fish recruitment.Chub mackerel Scomber japonicus is a commercially important pelagic fish widely distributed in the northwestern Pacific.Its catch showed large fluctuations with changes in distribution and migration under climate change and strong fishing.We determined the hatch dates and growth rates of young-of-the-year of chub mackerel through otolith microstructure using samples collected in the Oyashio water in autumn 2018.Results show that the ages of young chub mackerel ranged between 120 and 180 d,and the estimated hatch date lasted from midJanuary to late May with a peak from mid-March to mid-April.Average otolith daily increment width during the early life stages(from hatching to 25 d)showed an increasing trend.Chub mackerel grows slowly in the first 10 d,and then grows faster during the 10thto 25thd.Three groups with dissimilar growth histories and migration routes were identified using unsupervised random forest clustering analysis,but all eventually converge on the same nursery ground.The faster growth of young-of-the-year chub mackerel leads to better recruitment due to the hypothesis of growth-dependent mortality.Most chub mackerels hatched in March and April,the spawning period is longer and earlier,which could lead to strong year classes.These findings on population composition and life history traits of young-of-the-year of chub mackerel provide valuable information on its recruitment processes during the period of stock recovery.

Evaluation of Reference Genes for Quantitative Real-Time PCR Analysis in Manila Clam Ruditapes philippinarum Under Hypoxic Stress

Quantitative real-time PCR(qRT-PCR)has been widely used for gene expression analysis,and selection of reference genes is a key point to obtain accurate results.To find out optimal reference genes for qRT-PCR in Manila clam Ruditapes philippinarum in response to hypoxia,different tissues were used and compared to evaluate the stability of candidate reference genes under low oxygen stress(DO 0.5mgL^(−1) and DO 2.0mgL^(−1))and normal condition(DO 7.5mgL^(−1)).Seven candidate reference genes were selected to evaluate the stability of their expression levels.The reference genes were evaluated by Delta Ct,BestKeeper,NormFinder and geNorm,and then screened by RefFinder calculation.Under hypoxic stress of 0.5mgL^(−1),the most suitable reference gene for gill and hepatopancreas was RPL31,and the optimal reference genes for axe foot and adductor muscle were TUB and HIS,respectively.For hypoxic stress of 2.0mgL^(−1),the most stable reference gene for gill and hepatopancreas was RPL31,and the optimal reference genes for axe foot and adductor muscle were RPS23 and EF1A,respectively.At the normal condition,HIS and EF1A were identified as the optimal internal reference genes in gill and hepatopancreas respectively,and GFRP2 was the best internal reference gene for axe foot and adductor muscle.The present findings will provide important basis for the selection of reference genes for qRT-PCR analysis of gene expression level in bivalves under hypoxic stress,which might be helpful for the analysis of other molluscs too.

Contraction of Heat Shock Protein 70 Genes Uncovers Heat Adaptability of Ostrea denselamellosa

The milin oyster Ostrea denselamellosa is a live-bearing species with a sharp decline in the natural population.Unlike other oysters,O.denselamellosa lives in the subtidal zone and its adaptability to heat,salinity,etc.is different from most other oys-ters.Heat shock proteins 70(HSP70)are a family of conserved ubiquitously expressed heat shock proteins which are produced in re-sponse to stressful conditions,especially heat.In this study,we identified Hsp70 genes through bioinformatic analysis in five species of oyster.Among them,O.denselamellosa holds the fewest number of Hsp70 genes,which may be one of the reasons why O.den-selamellosa cannot tolerate high temperatures.The conserved motifs and gene structures of Hsp70B2 sub-family and other types of Hsp70 in O.denselamellosa were different from that of Hspa12 sub-family,which may be due to performing necessary multiple phy-siological functions.Transcription profile analysis for Hsp70 genes of O.denselamellosa indicated that gills play an important role in responding to multiple external challenges.In addition,synteny analysis of Hsp70 genes among O.denselamellosa,O.edulis and Cras-sostrea ariakensis showed that Hsp70 genes in genus of Ostrea genome might have evolved from a common ancestor with genus of Crassostrea.In short,our results lay the foundation for further investigation of the evolution of O.denselamellosa Hsp70 genes and heat adaptability.

Effects of Dietary Vitamin D_(3)on Growth Performance,Immune Status,and Calcium and Phosphorus Metabolism in Litopenaeus vannamei Under Different Salinity Conditions

Vitamin D_(3)(VD_(3)) plays a vital role in various physiological processes in addition to its role in regulating the homeostasis of calcium and phosphorus.The present study was conducted to investigate the effects of dietary VD_(3) on growth performance,immune status and the metabolism of calcium and phosphorus in Litopenaeus vannamei under the optimal salinity(25)and high salinity(35)conditions.Shrimp(2.1 g±0.1 g)were fed with experimental diets containing 0,1500,6000 and 12000IUkg^(−1)VD_(3) for 30 days under two salinity conditions respectively.The results showed that the growth performance and intestinal health were reduced,while oxidative stress was induced in the shrimp cultured at salinity 35 compared to those in the shrimp cultured at salinity 25.Interestingly,dietary supplementation of 1500 IU kg^(−1)VD_(3) at salinity 35 promoted the growth performance,improved the intestinal health,and enhanced the antioxidant capacity and non-specific immunity in shrimp.In addition,the content of inorganic phosphorus in shrimp serum was affected by dietary VD_(3) independent of salinity.In contrast,the expressions of calcium transport-related genes,including calmodulin and Ca2+-ATPase,were regulated by salinity,instead of dietary VD_(3).In conclusion,an appropriate supplementation of dietary VD_(3) under high-salt condition can promote the growth,alleviate intestinal inflammation,enhance antioxidant capacity and immunity,and contribute to phosphorus metabolism in L.vannamei.

Productive Traits and Triploid Rate Stability in Triploidy-Induced ‘Haida No. 2’ Strain of the Pacific Oyster, Crassostrea gigas

In order to evaluate the effects of triploidy induction on a selected strain‘Haida No.2’of the Pacific oyster Crassostrea gigas,which is characterized with golden shell color and high growth rate,the growth,survival rate and stability of triploid rate were analyzed at different development stages in the present study.Three different conditions inhibiting the release of polar body Ⅱ or polar body Ⅰ were tested:(A)Cytochalasin-B(CB),0.5mg L^(−1) at 10min post-insemination for 15 min;(B)CB,0.5mg L^(−1)at 15 min postinsemination for 20 min;and(C)CB,0.7mg L^(−1),at 15 min post-insemination for 20 min.The triploidy induction treatments significantly reduced the D-larvae and survival rates at the larvae stage but not at the juvenile and adult stages.Triploid rate dramatically decreased at the larval stage and did not significantly change at the juvenile and adult stages.Regarding the stability of the triploid rate,there was a significant difference between the three treatment groups.Larvae from the treatment A and control groups exhibited higher growth rates in shell height than those from the other two treatment groups at day 27.Triploid juveniles and adults from the treatment A group exhibited a higher wet weight than diploids from the control group and triploids from the other treatment groups.Triploidy induction did not affect the shell color of the progeny.The results obtained in the study demonstrate that triploidy induction has the potential to be used to increase the production of C.gigas variety‘Haida No.2’without modifying its golden shell color.

Characterization of Caspase Gene Family Members in Spotted Sea Bass(Lateolabrax maculatus)and Their Expression Profiles in Response to Vibrio harveyi Infection

The caspase gene family is a crucial gene cluster that regulates apoptosis which contribute to programmed cell death,cell proliferation and differentiation,and several immune responses.In our study,a complete set of 12 caspase genes were identified in spotted sea bass Lateolabrax maculatus.These genes were divided into three subfamilies:2 inflammatory caspases(casp-1 and casp-14-like),5 apoptosis initiators(casp-2,casp-8a,casp-8b,casp-9,and casp-10),and 5 apoptosis executioners(casp-3a,casp-3b,casp-3-like,casp-6,and casp-7).Their phylogenetic relationships,synteny and gene structures were systematically analyzed.Furthermore,the relative expression profiles of the caspase family members in the liver,intestine,head kidney,and spleen were measured by q PCR after infection with Vibrio harveyi.The results showed that the overall mRNA levels of the caspase genes were dramatically increased after V.harveyi infection,and the expression patterns varied among genes and tissues.More caspase genes underwent pronounced expression changes in the head kidney and spleen than in the liver or intestine,mainly after 48 h of the challenge.Specifically,casp-3a,casp-3b,casp-3-like,casp-6,casp-7,casp-8a,casp-8b,casp-10,and casp-14-like in the head kidney,and casp-3-like,casp-6,casp-7,and casp-14-like in the spleen,were the most responsive caspase genes which may contribute significantly to immune regulation in spotted sea bass.Additionally,the apoptosis level in head kidney and spleen after infection were examined using the Caspase assay.Our study provides a systemic overview of the caspase gene family in spotted sea bass after V.harveyi infection and lays a foundation for further deciphering the biological roles of these caspase genes.

Discordant patterns of genetic variation between mitochondrial and microsatellite markers in Acanthogobius ommaturus across the coastal areas of China

Acanthogobius ommaturus, which belongs to the family Gobiidae, is a euryhaline and demersal fish that is widely distributed in the coastal areas, harbors, and estuaries of China, D. P. R. Korea and Japan. In this study, the genetic diversity and genetic structure of five geographical populations of A. ommaturus was assessed using the mitochondrial hypervariable region gene and microsatellite markers. The results of the two genetic markers indicated that the A. ommaturus populations had a high level of genetic diversity. The mitochondrial marker detected weak genetic differentiation among populations, and the Neighbor-Joining tree showed that there was no obvious pedigree branches and geographic structure as well. However, population of Zhoushan showed significant genetic differentiation with other populations by microsatellite markers. The population of A.ommaturus has not experienced bottleneck effect recently. We speculated that the Pleistocene climate change and juvenile fish dispersal played an important role in the population differentiation of A. ommaturus.

Growth Performance of Complete Diallel Crosses Among New Varieties of Chinese Pearl Oyster Pinctada martensii

Pinctada martensii is an important shellfish for the production of sea pearls in China.To improve the growth performance of P.martensii,we successfully established three complete diallel crosses using three new varieties of P.martensii,which were named Haiyou No.1,Haixuan No.1,and Nanke No.1,to investigate the growth performance of each self-cross line and hybrid line in Beihai,Guangxi Zhuang Autonomous Region,China.Generally,high fertilization and hatching rates were observed in the experiment,suggesting that there was no sperm-egg recognition barrier among these three P.martensii varieties.The survival rate of the self-cross lines was relatively higher than that of hybrid lines,and♀‘Haixuan No.1’×♂‘Haiyou No.1’maintained a high survival rate during the grow-out stage.The shell length and width were affected by the genotypes as well as the interaction between the genotypes and environmental factors on day 400.The variances of the general combining abilities(GCA)and specific combining abilities(SCA)for shell length and width were the result of both additive genetic variance and nonadditive genetic variance.When Haixuan No.1 was the female parent and Nanke No.1 was the male parent,the GCA for shell length and width were both positive on day 400.The heterosis of Haixuan No.1 and Nanke No.1 in terms of shell length and width was positive.From the resulting combining ability and heterosis,♀‘Haixuan No.1’×♂‘Nanke No.1’was considered an ideal hatchery method to improve the growth performance of P.martensii.Our results therefore demonstrated that crossbreeding among these three new varieties can further improve the growth performance of P.martensii.

Molecular phylogenetics and population demographic history of Amphioctopus fangsiao,inferred from mitochondrial and microsatellite DNA markers

Amphioctopus fangsiao(Cephalopoda:Octopodidae)is an important commercial species in the coastal waters of China.In recent years,however,the resource of A.fangsiao have declined because of habitat destruction and overfishing.To analyze the genetic variations of A.fangsiao caused by the fluctuation of resources,the population genetic structure of nine sampling locations collected from the Bohai Sea to the South China Sea were investigated,using mtDNA COI fragments and microsatellite DNA.The results of F-statistics,AMOVA,STRUCTURE and PCA analyses showed three phylogeographic clades(Clades A,B and C),revealing limited genetic exchange between north and south populations.These clades diverged in 2.23(Clades A and B)and 3.67(Clades A,B and C)million years ago,during the dramatic environmental fluctuations,such as sea level and temperature changes,have exerted great influence on the survival distribution pattern of global organisms.Our results for low genetic connectivity among A.fangsiao populations provide insights into the development of management strategies,that is,to manage this species as separate management unit.