Six main mitochondrial DNA(mt DNA) lineages have been described in minnow(Zacco platypus) samples obtained from northern, western and southern China. Perdices et al.(2004) predicted that further sampling of other trib...Six main mitochondrial DNA(mt DNA) lineages have been described in minnow(Zacco platypus) samples obtained from northern, western and southern China. Perdices et al.(2004) predicted that further sampling of other tributaries might discover more lineages of this species. In this study, we collected 26 Zacco platypus individuals in the Huangshan area of eastern China and determined the cytochrome b(cytb) sequence variations. Combined with reported data in Gen Bank, we identified ten matrilines(Zacco A-J) in a total of 169 samples, with relatively high molecular divergence found among them. The Huangshan population had the greatest genetic variation among all sampled regions and hosted six of the ten matrilines. Our results highlight the significance of the Huangshan area for the conservation of Zacco platypus.展开更多
Piwi-interacting RNA(piR NA) plays an important role in the gonadal development and maintenance of Teleostei. In this study, pi RNA libraries derived from the adult gonads of Japanese flounder(Paralichthys olivaceus) ...Piwi-interacting RNA(piR NA) plays an important role in the gonadal development and maintenance of Teleostei. In this study, pi RNA libraries derived from the adult gonads of Japanese flounder(Paralichthys olivaceus) were generated using next-generation sequencing technology. Using zebrafish piR NAs as a reference, 5 865 unique candidate pi RNAs were identified; 289 candidate piR NA clusters(PRCs) were generated from the above pi RNAs. Among the isolated candidate PRCs, a total of 38 ovary-specific, 45 ovary-bias, 24 testis-specific, and 131 testis-bias PRCs were found. The relative expression levels of seven PRCs were validated through quantitative reverse transcription-polymerase chain reaction. The results of this study will help facilitate exploration of the development and maintenance of the phenotypic sex mechanism in P. olivaceus.展开更多
Rotifer community structure was investigated in Yangcheng Lake in 2008. Dominant species and species diversity indices were determined and QB/T was used to assess water quality. The annual average density and biomass ...Rotifer community structure was investigated in Yangcheng Lake in 2008. Dominant species and species diversity indices were determined and QB/T was used to assess water quality. The annual average density and biomass of the rotifers in Yangcheng Lake was 2 894 ± 1 006 ind./L and 12.47 ± 10.28 mg/L, respectively. The highest densities were observed in the western portion of the lake in March, but the highest biomass occurred in inflowing creeks in September. Within a year of monitoring in Yangcheng Lake, 93 species were identified and the dominant species were found to be Polyarthra trigla, Brachionus angularis, Keratella cochlearis, Keratella valga, Brachionus calyciflorus, and Filinia major. Of the species recorded, 75 were pollution indicator species. Density and biomass exhibited significant positive correlations with water temperature (R = 0.209, P = 0.003; R = 0.446, P = 0.000), but the peak density showed two lags in response to chl a. According to the Jaccard similarity index (SJ), the greatest similarity among dominant species occurred between creeks and the eastern part of the lake. The annual average Shannon-Wiener diversity index H', Margalef richness index D and Pielou evenness index J were 1.96 ± 0.34, 1.61 ± 0.50 and 0.77 ± 0.10, respectively. In all four areas of Yangcheng Lake, β- mesosaprobic species comprised the largest share of pollution indicator species. These data suggest that Yangcheng Lake is mesosaprobic.展开更多
The single and synergistic effects of dietary arabinoxylan-oligosaccharide(AXOS)and inulin at different doses(2,4,and 8 mg/g diet)on survival,growth performance,gut microbiota,and immune response in Pacific white shri...The single and synergistic effects of dietary arabinoxylan-oligosaccharide(AXOS)and inulin at different doses(2,4,and 8 mg/g diet)on survival,growth performance,gut microbiota,and immune response in Pacific white shrimp Litopenaeus vannamei were assessed.Singular application of either inulin or AXOS at doses of 4 mg/g diet showed the most stimulatory effects on the growth rate and gene expression levels of chitinase,cathepsin L,chymotrypsin,and extracellular signal-regulated kinase(ERK)in each single prebiotic feeding trial.Compared with single prebiotic treatments,simultaneous application of AXOS and inulin at 4 mg/g diet enhanced remarkably the growth parameters of shrimp and the expression of related genes(chitinase,cathepsin L,chymotrypsin,ERK,myeloid differentiation factor 88,and phenoloxidase)after 8-week feeding(P<0.05).Additionally,gut microbiota analysis indicated the dietary supplementation with combined prebiotics increased significantly the bacterial community richness and relative abundance of Bacillus Pseudomonas Bacteriovorax,and Lactobacillus,and reduced the abundance of Vibrio Rhodococcus,and Photobacterium in the digestive tract of L vannamei.Compared with the single prebiotic treatment and the control,combined prebiotics supplementation boosted notably the survival rate and expression levels of immune-related genes in shrimp infected with Vibrio alginolyticus or white spot syndrome virus.Therefore,simultaneous application of AXOS and inulin shall have a great potential of dietary supplement in the culture of L.vannamei.展开更多
Stable isotopes are increasingly used to investigate seasonal migrations of aquatic organisms. This study employed stable isotopes(δ^(13) and δ15N) for Coilia nasus from the lower Yangtze River and the adjacent East...Stable isotopes are increasingly used to investigate seasonal migrations of aquatic organisms. This study employed stable isotopes(δ^(13) and δ15N) for Coilia nasus from the lower Yangtze River and the adjacent East China Sea to distinguish different ecotypic groups, ascertain trophic nutrition positions, and reflect environmental influences on C. nasus. δ^(13)C signatures of C. nasus sampled from Zhoushan(ZS), Chongming(CM), and Jingjiang(JJ) waters were significantly higher than those from the Poyang Lake(PYL)(P < 0.05). By contrast, δ^(15)Nsignatures of C. nasus in ZS, CM, and JJ groups were significantly lower than those in PYL group(P < 0.05). Basing on δ^(13) and δ^(15)Nsignatures, we could distinguish anadromous(ZS, CM, and JJ) and non-anadromous(PYL) groups. The trophic level(TL) of anadromous C. nasus ranged from 2.90 to 3.04, whereas that of non-anadromous C. nasus was 4.38. C. nasus occupied the middle and top nutrition positions in the marine and Poyang Lake food webs, respectively. C. nasus in Poyang Lake were significantly more enriched in δ^(15)N but depleted in δ^(13), suggesting that anthropogenic nutrient inputs and terrigenous organic carbon are important to the Poyang Lake food web. This study is the first to apply δ^(15)Nand δ^(13) to population assignment studies of C. nasus in the Yangtze River and its affiliated waters. Analysis of stable isotopes(δ^(15)Nand δ^(13)) is shown to be a useful tool for discriminating anadromous and non-anadromous C. nasus.展开更多
It is suggested that Δ6 fatty acid desaturase(FAD) plays a critical role in the biosynthesis of polyunsaturated fatty acids in plants and microalgae. But why does it adapt to the changed environments such as nitrogen...It is suggested that Δ6 fatty acid desaturase(FAD) plays a critical role in the biosynthesis of polyunsaturated fatty acids in plants and microalgae. But why does it adapt to the changed environments such as nitrogen starvation is seldom understood. One Δ6 FAD gene( MiD6 fad) from an arachidonic acidrich microalga M yrmecia incisa Reisigl(Chlorophyta) was first heterologously expressed in S accharomyces cerevisiae for the identification of function. The fatty acid profile of transgenic yeast detected by gas chromatography-mass spectrometry illustrated that the enzyme MiD6 FAD could convert linoleic and ?-linolenic acids to γ-linolenic and stearidonic acids, respectively, demonstrating that M iD6 fad encoded a Δ6 FAD. A 1 965-bp fragment of the cloned 2 347-bp 5′-upstream region of M iD6 fad was next subcloned and fused upstream with green fluorescent protein(GFP) gene to replace the GAL1 promoter of the vector pYES2. The generated construct was transformed into S. cerevisiae for function determination. Confocal microscopic images of the transformed line illustrated that this inserted fragment could drive GFP expression, which was further verified by fluorescence intensity quantification and Western blot analysis using antiGFP antibody. The conversion efficiency(approximately 2%-3%) of MiD6 FAD was much lower than the reported ? 3 FAD and Δ6 elongase in this microalga, suggesting that MiD6 FAD catalysed the possible ratelimiting step for ArA biosynthesis. The presence of several putative c is-acting regulatory elements in this identified promoter sheds new light on the regulation mechanism research of Δ6 FAD transcription for the ArA production in M. incisa in changing environmental factors.展开更多
Invasive species generally possess unique characteristics that allow them to survive the invasion process in order to establish and spread in new habitats. Successful invaders must resist both physical and physiologic...Invasive species generally possess unique characteristics that allow them to survive the invasion process in order to establish and spread in new habitats. Successful invaders must resist both physical and physiological stresses associated with the changing environment. A common littoral barnacle, Chthamalus challengeri Hoek, 1883(Crustacea, Cirripedia), which is native to Japan, South Korea and northern China, has become established in the high-littoral zone adjacent to Yangshan Port, Shanghai, China. A comparison of the morphology of Chthamlus species from Zhoushan archipelago with previous description indicates the occurrence of C. challengeri. The new immigrant becomes a dominant species in certain high-intertidal habitats of the adjacent area to of Yangshan Port. C. challengeri was found in part of sampling sites in Zhoushan in 2010; however, it dispersed to all the eleven sampling sites in 2012. Densities of C.challengeri had increased over 10 times in the last 2 years, with the highest mean value reaching 39533 ± 6243 ind. m-2 in the new habitat. The specific ratios of both operculum area(Sa) to base area(SA) and average height of parietal plates(H) to length of base(L) revealed that C. challengeri displays morphological changes to resist stronger currents in the new habitats for invasion.展开更多
It has been reported that there was a linkage of 5S rRNA gene to 18S-5.8S-25S rRNA gene in a few of species in Ochrophyta.In regard to the usual two positions of linked 5S rDNA to the 3′end of 25S rDNA,two pairs of p...It has been reported that there was a linkage of 5S rRNA gene to 18S-5.8S-25S rRNA gene in a few of species in Ochrophyta.In regard to the usual two positions of linked 5S rDNA to the 3′end of 25S rDNA,two pairs of primers were designed for amplification to verify this linkage of two genes in a kelp cultivar of Saccharina japonica,one of species in Ochrophyta.This result supplemented the previous report that 5S rDNA was unlinked to 25S rDNA in this kelp.In order to simultaneously visualize this unlinkage of two genes,dual-color fluorescence in situ hybridization(FISH)technique was applied to the cytogenetics of S.japonica.Dual-color FISH images showed that two and four hybridization signals were present in the kelp gametophyte and sporophyte,respectively,metaphase nuclei hybridized simultaneously with the labeled probes of 18S rDNA and 5S rDNA.Both haploid and diploid karyotypes in decreasing length of chromosomes showed that 18S-5.8S-25S rDNA was localized at the interstitial region of Chromosome 23,whereas 5S rDNA resided at the sub-telomeric region of Chromosome 27.These karyotypes suggested that the kelp nuclear genome had only one locus of each rRNA gene,and their loci on different chromosomes indicated the physical unlinkage of 5S rDNA to 18S-5.8S-25S rDNA in this kelp.Therefore,dual-color FISH seems to be a powerful technique for the discrimination and pairing of chromosomes featured in both small size and nearly identical shape in S.japonica.展开更多
Interleukin-21(IL-21)is produced mainly by activated CD4^(+)T cells and promotes the proliferation,survival and differentiation of immune cells.This study characterised a high affinity binding receptor of IL-21(IL-21 ...Interleukin-21(IL-21)is produced mainly by activated CD4^(+)T cells and promotes the proliferation,survival and differentiation of immune cells.This study characterised a high affinity binding receptor of IL-21(IL-21 Rα)from grass carp(Ctenopharyngodon idella,Ci)and analyzed the expression of CiIL-21 and CiIL-21 Rα.It has been shown that they were modulated by PAMPs and during infection with bacterial and viral pathogens.Recombinant CiIL-21 protein produced in bacteria was active in inducing the expression of genes involved in Th responses.A monoclonal CiIL-21 antibody was generated against the rCiIL-21 and showed good reactive specificity with the rCiIL-21 proteins expressed in E.coli cells,and HEK293 T cells and the native protein from PHA-stimulated kidney leukocytes.In addition,the IL-21-producing cells detected by the CiIL-21 monoclonal antibody were found to be increased to in the kidney,spleen and hindgut after infection with Flavobacterium columnare.Our data indicate that IL-21 is involved in the regulation of immune response to bacterial and viral infection.展开更多
Miiuy croaker,Miichthys miiuy is an ecologically important teleost species which is widely distributed in southeast coast of China.In this study,we present a chromosomal-scale genome assembly of the miiuy croaker whic...Miiuy croaker,Miichthys miiuy is an ecologically important teleost species which is widely distributed in southeast coast of China.In this study,we present a chromosomal-scale genome assembly of the miiuy croaker which is an important Sciaenidae fish and economical species.We adopted Oxford Nanopore and Hi-C sequencing techniques to achieve an assembly with high accuracy and completeness.The investigation of genome characteristic and functional features may provide insights into the study of phylogenetic diversity of miiuy croaker.This study can also be beneficial to improve molecular assisted breeding techniques.Moreover,it can be a great resource to better conduct further study of other sciaenids.展开更多
CD4 T helper cells are an important group of cells in the immune system of vertebrates and express CD4 receptor on the cell surface.In mammals,the CD4 receptor is encoded by a single copy gene,whilst in fish,two copie...CD4 T helper cells are an important group of cells in the immune system of vertebrates and express CD4 receptor on the cell surface.In mammals,the CD4 receptor is encoded by a single copy gene,whilst in fish,two copies of cd4 genes,namely cd4-1 and cd4-2,are found.In this study,the ectodomains of grass carp(Ctenopharyngodon idella,Ci)CD4-1 and CD4-2 were expressed in the E.coli cells and used to generate monoclonal antibodies in mice.Western blotting,confocal microscopy and flow cytometry were performed to characterize the monoclonal antibodies.It has been shown that the CiCD4-1 and CiCD4-2 monoclonal antibodies had good specificity to react with the recombinant ectodomains of CiCD4-1 and CiCD4-2 expressed in the CHO-S cells and the native CD4 molecules of grass carp.The CiCD4-1 monoclonal antibody did not recognize CiCD4-2 and verse versa.In addition,the CD4-1 and CD4-2 monoclonal antibodies specifically recognized the CD4-1 and CD4-2 receptors expressed in the HEK293 cells and native molecules of fish cells.Further,the percentages of lymphocytes in immune tissues of healthy fish were analyzed by flow cytometry.It was found that 17.6%of lymphocytes were CD4-1^(+)cells and 22.5%were CD4-2^(+)in the head kidney.In the spleen,13.1%of lymphocytes were CD4-1^(+)and 18.6%were CD4-2^(+)while 7.3%of blood lymphocytes were CD4-1^(+)cells and 8.8%were CD4-2^(+)cells.The availability of the CD4-1 and CD4-2 monoclonal antibodies provides antibody-based tools for further elucidation of the functions of CD4 T cells in grass carp.展开更多
Cyanobacterial blooms in eutrophic freshwater systems are a worldwide problem, creating adverse effects for many aquatic organisms by producing toxic microcystins and deteriorating water quality. In this study, microc...Cyanobacterial blooms in eutrophic freshwater systems are a worldwide problem, creating adverse effects for many aquatic organisms by producing toxic microcystins and deteriorating water quality. In this study, microcystins (MCs) in Microcystis aeruginosa, and Daphnia magna exposed to M. aeruginosa, were analyzed by HPLC-MS, and the effects of M. aeruginosa on D. magna were investigated. When D. magna was exposed to M. aeruginosa for more than 2 h, Microcystin-LR (MC-LR) was detected. When exposed to 1.5 × 106, 3 × 106, 0.75 × 107, and 1.5 × 107 cell/mL of M. aeruginosa for 96 h, average survival of D. magna for treatments were 23.33%, 33.33%, 13.33%, 16.67%, respectively, which were significantly lower than the average 100% survival in the control group (P < 0.05). The adverse effects of M. aeruginosa on body length, time for the first brood, brood numbers, gross fecundity, lifespan, and population growth of D. magna were density-dependent. These results suggest that the occurrence of M. aeruginosa blooms could strongly inhibit the population growth of D. magna through depression of survival, individual growth and gross fecundity. In the most serious situations, M. aeruginosa blooms could undermine the food web by eliminating filter-feeding zooplankton, which would destroy the ecological balance of aquaculture water bodies.展开更多
The nucleotide sequence of the mitochondrial genome of the solecurtidae Bivalvia mollusca Sinonovacula constricta(GenBank accession number EU880278) has been determined and is reported here.We determined the complete ...The nucleotide sequence of the mitochondrial genome of the solecurtidae Bivalvia mollusca Sinonovacula constricta(GenBank accession number EU880278) has been determined and is reported here.We determined the complete mitochondrial genome sequence using long-PCR and Shot Gun Sequencing.Contained within the 17 225 base pairs(bp) are the two ribosomal RNA genes and 12 protein coding genes typical of metazoan mitochondrial genomes.The S.constricta mitochondrial DNA(mtDNA) did not contain a gene for atp8,similar to the mtDNA of Crassostrea virginica,Crassostrea giga and Mytilus edulis.The S.constricta mtDNA is 67.0% A+T(A 25.9%,C 10.5%,G 22.5%,and T 41.1%).This value is higher than that for many invertebrate mitochondrial genomes.Only 19 putative tRNA genes are present in S.constricta and 27 noncoding regions,of which two are large in size.The trnE and trnW genes as well as a second trnS were absent in S.constricta.The gene arrangement of S.constricta is different from the other Bivalvia genomes.展开更多
Pituitary adenylate cyclase activating polypeptide(PACAP)and growth hormone-releasing hormone(GHRH)play important roles in the GH/IGF growth axis in fishes.To determine whether epigenetic change is involved in the reg...Pituitary adenylate cyclase activating polypeptide(PACAP)and growth hormone-releasing hormone(GHRH)play important roles in the GH/IGF growth axis in fishes.To determine whether epigenetic change is involved in the regulation of pacap and ghrh responses to low salinity stress in Cynoglossus semilaevis,the correlation between growth traits,DNA methylation status and gene expression level in low salinity(15,S15)and optimal salinity(30,S30)at day 7(D7)and day 60(D60)were analyzed.Results showed that exposure to low salinity for 60 days attenuated C.semilaevis growth rate.Under low salinity,DNA methylation level of pacap promoter increased in females and decreased in males at day 7,but inverted at day 60.Additionally,pacap expression was up-regulated in both males and females.The pacap promoter methylation level was higher and expression level was lower in female than in male.The results suggest that pacap promoter methylation level is negatively correlated to mRNA level and positively correlated to body weight,while gene expression level is negatively related with body weight.With the decrease of salinity,DNA methylation level of ghrh promoter and exon1,as well as its gene expression displayed minor changes.Overall,pacap gene seems to play an important role in fish growth,contributing to female growth superiority,while ghrh gene seems not pertinent under salinity stress.The results indicate that low salinity potentially affects fish growth through regulating DNA methylation in pacap promoter.This study expands the understanding of the molecular mechanism of how salinity modulates fish growth from the epigenetic perspective.展开更多
The population of tri-spine horseshoe crabs(HSCs)Tachypleus tridentatus has sharply decreased in recent decades.Domestication and manual releasing are now used to recover it.In order to accelerate the growth of T.trid...The population of tri-spine horseshoe crabs(HSCs)Tachypleus tridentatus has sharply decreased in recent decades.Domestication and manual releasing are now used to recover it.In order to accelerate the growth of T.tridentatus in indoor environment,in this study,gut microbiota of 2nd instar T.tridentatus was supposed to be improved by putting probiotics in water environment and thus promoted the health and growth of the host.Using 16S rRNA sequencing of gut samples,we examined the intestinal microbiome of juvenile HSCs and studied the effect of probiotics.Results showed that there was no significant difference between groups fed with high and low concentrations(1×10^(7),1×10^(12)CFU mL^(−1))of Bacillus subtilis and high concentration(1×10^(12)CFU mL^(−1))of Lactobacillus,and no significant difference compared with the control group,but the diversity of Lactobacillus group with low concentration(1×10^(7)CFU mL^(−1))was significantly higher than those of other groups.Microbial-mediated functions predicted by PICRUSt showed that‘Cellular Processes’,‘Metabolism’and‘Organismal Systems’pathways significantly increased in groups of Bacillus subtilis and Lactobacillus group with low concentration.The results showed that two types of probiotics with different concentrations have other effects on the structure,richness and function of intestinal microbes.By searching the literature and conducting pre-experiments,we forecasted the short time of breeding under the probiotic environment may not have much effect.The present study for the first time investigated the impact of probiotics on gut microbiota of juvenile HSCs and found them may benefit for the growth and health of HSCs.展开更多
A cell line,termed ZFIN,was established from the caudal fin of zebrafish and was shown to be susceptible to spring viremia of carp virus(SVCV).The ZFIN cells are epithelial like cells and have a moderate plasmid trans...A cell line,termed ZFIN,was established from the caudal fin of zebrafish and was shown to be susceptible to spring viremia of carp virus(SVCV).The ZFIN cells are epithelial like cells and have a moderate plasmid transfection efficiency of 13.9%.Using an RNA-seq approach,differentially expressed genes(DEGs)regulated by SVCV were identified.Infection of SVCV gave rise to 3931 DEGs and up-regulated DEGs were mostly enriched into the biological regulation and cellular processes,among which pathways for the type I interferon signaling and the response to exogenous dsRNA were the top two GO terms.Several KEGG signaling pathways including TLR signaling pathway,RLR receptor signaling pathway,cytosolic DNA-sensing pathway,NLR signaling pathway,cytokine-cytokine receptor interaction and ferroptosis were significantly enriched.Antiviral genes including ifnφ1,isg15 and mx were significantly up-regulated.In addition,key DEGs involved in autophagy were identified.The results indicate that the ZFIN cell line provides a useful in vitro tool for study on the gene functions and cellular responses to viral infection in fish.展开更多
Lysine-specific demethylase 4A(KDM4A)catalyzes demethylation of histone lysine residues,which regulates chromatin state and transcription.In drosophila and mice,KDM4A plays an important role in multiple biological pro...Lysine-specific demethylase 4A(KDM4A)catalyzes demethylation of histone lysine residues,which regulates chromatin state and transcription.In drosophila and mice,KDM4A plays an important role in multiple biological processes including development,aging,metabolism,and immunity,however the functions of KDM4A in fish are still unclear.There are two copies of the kdm4a gene in zebrafish,namely kdm4aa and kdm4ab,kdm4aa was edited using CRISPR/Cas9 technology in the present study,then homozygous kdm4aa mutants(kdm4aa^(-/-))were obtained,and loss of kdm4aa was confirmed by sequencing and increased H3K9me3.Whole-mount in situ hybridization showed that kdm4aa is widely expressed during the embryonic development of zebrafish.Compared with WT zebrafish,kdm4aa^(-/-)zebrafish showed no significant difference in gamete formation and fertilization,but the survival rate of kdm4aa^(-/-)embryos dramatically reduced to 21%at 26 hpf.Further observation showed that about 80%of survived kdm4aa^(-/-)zebrafish experienced disruption in stripe formation,and 10%of survived kdm4aa^(-/-)zebrafish underwent vertebral malformation.Alizarin red S staining demonstrated the abnormal spinal development in kdm4aa^(-/-)zebrafish.These results indicated that kdm4aa is required for normal embryonic development of zebrafish,loss of kdm4aa function leads to decreased survival during the early stages of zebrafish development and morphological variation in adult zebrafish.展开更多
Noncoding RNA(ncRNA)is an important regulatory factor that plays a major role in innate immunity.However,most studies on ncRNA have focused on mammals,resulting in a knowledge gap on nc RNA in lower vertebrates such a...Noncoding RNA(ncRNA)is an important regulatory factor that plays a major role in innate immunity.However,most studies on ncRNA have focused on mammals,resulting in a knowledge gap on nc RNA in lower vertebrates such as teleost fish.In this study,we identified a new long noncoding RNA(lncRNA),termed TAK1-related lnc RNA(TARL),which can play a positive role in the antibacterial immunity of Miichthys miiuy to Vibrio anguillarum and V.harveyi.We also found a novel micro RNA miR-2188-3p that could target TAK1 and inhibit the host antibacterial response and promote bacterial escape.We further found that the antibacterial effect inhibited by miR-2188-3p could be reversed with TARL.Moreover,V.anguillarum and V.harveyi are the two most susceptible Gram-negative pathogens of aquaculture fish,and the economic losses caused by these two bacteria are immeasurable every year.This study is the first to report on the ability of lnc RNA to prevent the escape of V.anguillarum and V.harveyi in fish through the competing endogenous RNA(ceRNA)mechanism.Our results not only elucidate the ce RNA mechanism of the lncRNA in antibacterial immune responses but also provide new insights into the impact of lnc RNA on host immunity and bacterial escape.展开更多
Activin A receptor,type II(Acvr2)is a member of the transforming growth factor beta receptor family and can function as a negative regulator of skeletal muscle mass.Acvr2 plays an important role in regulating muscle d...Activin A receptor,type II(Acvr2)is a member of the transforming growth factor beta receptor family and can function as a negative regulator of skeletal muscle mass.Acvr2 plays an important role in regulating muscle development that can inhibit skeletal muscle growth in mice.However,there is very little research reported on the function of acvr2 in muscle development of teleost.In this study,we analyzed the effect of acvr2aa and acvr2ba on muscle development in zebrafish.Growth rates of WT and acvr2a^(-/-b-/-)were measured from juvenile stage to adult stage.In addition,effects of acvr2 on skeletal muscle were tested in histological,protein and molecular levels.As a result,acvr2a^(-/-b-/-)exhibited a wider body trunk than WT and showed a significant increase in body weight and width from two months old.Histological analysis of skeletal muscle indicated that the size of muscle fiber in acvr2a^(-/-b-/-)(female:1809±123μm^(2);male:2261±130μm^(2))was larger than that in WT(709.8±49μm^(2);815±53μm^(2)).In addition,western blot of fast MyLc protein showed the protein synthesis of acvr2a^(-/-b-/-)are increased.Besides,Histological analysis of heart showed the ventricle area is aslo increased in acvr2a^(-/-b-/-).Our results demonstrated acvr2 attends the development of muscle fiber and will cause muscle hypertrophy when they were knocked out in zebrafish.In conclusion,acvr2 in zebrafish can control the development of muscle fibers during posthatch growth.展开更多
As an important seedling source,monospores closely associate with yields in nori farming.However,the molecular mechanism underlying differences in monospore production for different strains remains unknown.Comparative...As an important seedling source,monospores closely associate with yields in nori farming.However,the molecular mechanism underlying differences in monospore production for different strains remains unknown.Comparative transcriptome analysis was performed to examine gene expression differences between the spore abundant wild-type strain(WT)and spore deficient mutant(Y1)of Pyropia chauhanii.The WT strain that produces monospores in abundance exhibited more differentially expressed genes(DEGs)in both number and higher fold-changes than the Y1 strain incapable of producing monospores,indicating that the specific regulation of genes is involved in monospore production.Three lists of DEGs were obtained between the two strains using intersection and displayed in Venn diagram:one expressed only in WT strain,another expressed only in Y1 strain,and the third shared in both strains.DEGs annotated as homologous genes of Arabidopsis thaliana in these 3 lists were curated for online functional enrichment analysis on Metascape website.Gene regulatory networks of WT were functionally enriched in the processing,proteolysis,and transport of proteins,especially within the small GTPase protein family,which might be account for the monospore production ability,whereas Y1 were functionally enriched in the metabolism of essential substance and utilization of indispensable energy,which might be account for the rapid growth of blades.We found the differentially enriched gene regulatory networks between strains might be the intrinsic mechanisms of the different monospore production traits.These findings provide novel insights into the genes and regulatory networks associated with monospore production abilities,which are essential for developing accurate breeding technologies for optimal release of monospores and increase of total nori production.展开更多
基金Foundation items： This research was funded by the National Natural Science Foundation of China （NSFC 30870290, 31071891 and 31471968）ACKNOWLEDGEMENTS We extend our thanks to Dr. Robert W. MURPHY for his valuable comments on this manuscript. We thank Jin-Min CHEN （Yunnan University）, Bao-Lin ZHANG （Kunming Institute of Zoology, CAS） and Li-Fang PENG （Nanjing Forestry University） for their help in data processing. We further thank Dian- Cheng YANG （Nanjing Forestry University） and Jun-Sheng CUI （Anhui Agricultural University） for sampling.
文摘Six main mitochondrial DNA(mt DNA) lineages have been described in minnow(Zacco platypus) samples obtained from northern, western and southern China. Perdices et al.(2004) predicted that further sampling of other tributaries might discover more lineages of this species. In this study, we collected 26 Zacco platypus individuals in the Huangshan area of eastern China and determined the cytochrome b(cytb) sequence variations. Combined with reported data in Gen Bank, we identified ten matrilines(Zacco A-J) in a total of 169 samples, with relatively high molecular divergence found among them. The Huangshan population had the greatest genetic variation among all sampled regions and hosted six of the ten matrilines. Our results highlight the significance of the Huangshan area for the conservation of Zacco platypus.
基金supported by grants from the China Agriculture Research System(CARS-49)National Natural Science Foundation of China(31372520)Shanghai Collaborate Innovation Center for Aquatic Animal Genetics and Breeding(ZF1206)
文摘Piwi-interacting RNA(piR NA) plays an important role in the gonadal development and maintenance of Teleostei. In this study, pi RNA libraries derived from the adult gonads of Japanese flounder(Paralichthys olivaceus) were generated using next-generation sequencing technology. Using zebrafish piR NAs as a reference, 5 865 unique candidate pi RNAs were identified; 289 candidate piR NA clusters(PRCs) were generated from the above pi RNAs. Among the isolated candidate PRCs, a total of 38 ovary-specific, 45 ovary-bias, 24 testis-specific, and 131 testis-bias PRCs were found. The relative expression levels of seven PRCs were validated through quantitative reverse transcription-polymerase chain reaction. The results of this study will help facilitate exploration of the development and maintenance of the phenotypic sex mechanism in P. olivaceus.
基金Supported by the Scientific Special Fund of Commonweal Industry (Agriculture) of Finance Ministry (No. ayhyzx07-045)the Key Discipline Projects of Shanghai (No. S30701)the Doctor Startup Projects of Shanghai Ocean University
文摘Rotifer community structure was investigated in Yangcheng Lake in 2008. Dominant species and species diversity indices were determined and QB/T was used to assess water quality. The annual average density and biomass of the rotifers in Yangcheng Lake was 2 894 ± 1 006 ind./L and 12.47 ± 10.28 mg/L, respectively. The highest densities were observed in the western portion of the lake in March, but the highest biomass occurred in inflowing creeks in September. Within a year of monitoring in Yangcheng Lake, 93 species were identified and the dominant species were found to be Polyarthra trigla, Brachionus angularis, Keratella cochlearis, Keratella valga, Brachionus calyciflorus, and Filinia major. Of the species recorded, 75 were pollution indicator species. Density and biomass exhibited significant positive correlations with water temperature (R = 0.209, P = 0.003; R = 0.446, P = 0.000), but the peak density showed two lags in response to chl a. According to the Jaccard similarity index (SJ), the greatest similarity among dominant species occurred between creeks and the eastern part of the lake. The annual average Shannon-Wiener diversity index H', Margalef richness index D and Pielou evenness index J were 1.96 ± 0.34, 1.61 ± 0.50 and 0.77 ± 0.10, respectively. In all four areas of Yangcheng Lake, β- mesosaprobic species comprised the largest share of pollution indicator species. These data suggest that Yangcheng Lake is mesosaprobic.
基金Supported by the Shanghai Agriculture Applied Technology Development Program,China(Nos.G2014070111,201902080010F01111)。
文摘The single and synergistic effects of dietary arabinoxylan-oligosaccharide(AXOS)and inulin at different doses(2,4,and 8 mg/g diet)on survival,growth performance,gut microbiota,and immune response in Pacific white shrimp Litopenaeus vannamei were assessed.Singular application of either inulin or AXOS at doses of 4 mg/g diet showed the most stimulatory effects on the growth rate and gene expression levels of chitinase,cathepsin L,chymotrypsin,and extracellular signal-regulated kinase(ERK)in each single prebiotic feeding trial.Compared with single prebiotic treatments,simultaneous application of AXOS and inulin at 4 mg/g diet enhanced remarkably the growth parameters of shrimp and the expression of related genes(chitinase,cathepsin L,chymotrypsin,ERK,myeloid differentiation factor 88,and phenoloxidase)after 8-week feeding(P<0.05).Additionally,gut microbiota analysis indicated the dietary supplementation with combined prebiotics increased significantly the bacterial community richness and relative abundance of Bacillus Pseudomonas Bacteriovorax,and Lactobacillus,and reduced the abundance of Vibrio Rhodococcus,and Photobacterium in the digestive tract of L vannamei.Compared with the single prebiotic treatment and the control,combined prebiotics supplementation boosted notably the survival rate and expression levels of immune-related genes in shrimp infected with Vibrio alginolyticus or white spot syndrome virus.Therefore,simultaneous application of AXOS and inulin shall have a great potential of dietary supplement in the culture of L.vannamei.
基金financially supported by the Special Fund for Agro-scientific Research in the Public-Interest(Grant No.201203065)the National Natural Science Foundation of China(Nos.31172407+1 种基金1472280)the Specialized Research Fund for the Doctoral Program of Higher Education(No.20123104110006)
文摘Stable isotopes are increasingly used to investigate seasonal migrations of aquatic organisms. This study employed stable isotopes(δ^(13) and δ15N) for Coilia nasus from the lower Yangtze River and the adjacent East China Sea to distinguish different ecotypic groups, ascertain trophic nutrition positions, and reflect environmental influences on C. nasus. δ^(13)C signatures of C. nasus sampled from Zhoushan(ZS), Chongming(CM), and Jingjiang(JJ) waters were significantly higher than those from the Poyang Lake(PYL)(P < 0.05). By contrast, δ^(15)Nsignatures of C. nasus in ZS, CM, and JJ groups were significantly lower than those in PYL group(P < 0.05). Basing on δ^(13) and δ^(15)Nsignatures, we could distinguish anadromous(ZS, CM, and JJ) and non-anadromous(PYL) groups. The trophic level(TL) of anadromous C. nasus ranged from 2.90 to 3.04, whereas that of non-anadromous C. nasus was 4.38. C. nasus occupied the middle and top nutrition positions in the marine and Poyang Lake food webs, respectively. C. nasus in Poyang Lake were significantly more enriched in δ^(15)N but depleted in δ^(13), suggesting that anthropogenic nutrient inputs and terrigenous organic carbon are important to the Poyang Lake food web. This study is the first to apply δ^(15)Nand δ^(13) to population assignment studies of C. nasus in the Yangtze River and its affiliated waters. Analysis of stable isotopes(δ^(15)Nand δ^(13)) is shown to be a useful tool for discriminating anadromous and non-anadromous C. nasus.
基金Supported by the National Natural Science Foundation of China(No.31172389)the Special Project of Marine Renewable Energy from the State Oceanic Administration(No.SHME2011SW02)the Shanghai Universities Peak Discipline Project of Aquaculture
文摘It is suggested that Δ6 fatty acid desaturase(FAD) plays a critical role in the biosynthesis of polyunsaturated fatty acids in plants and microalgae. But why does it adapt to the changed environments such as nitrogen starvation is seldom understood. One Δ6 FAD gene( MiD6 fad) from an arachidonic acidrich microalga M yrmecia incisa Reisigl(Chlorophyta) was first heterologously expressed in S accharomyces cerevisiae for the identification of function. The fatty acid profile of transgenic yeast detected by gas chromatography-mass spectrometry illustrated that the enzyme MiD6 FAD could convert linoleic and ?-linolenic acids to γ-linolenic and stearidonic acids, respectively, demonstrating that M iD6 fad encoded a Δ6 FAD. A 1 965-bp fragment of the cloned 2 347-bp 5′-upstream region of M iD6 fad was next subcloned and fused upstream with green fluorescent protein(GFP) gene to replace the GAL1 promoter of the vector pYES2. The generated construct was transformed into S. cerevisiae for function determination. Confocal microscopic images of the transformed line illustrated that this inserted fragment could drive GFP expression, which was further verified by fluorescence intensity quantification and Western blot analysis using antiGFP antibody. The conversion efficiency(approximately 2%-3%) of MiD6 FAD was much lower than the reported ? 3 FAD and Δ6 elongase in this microalga, suggesting that MiD6 FAD catalysed the possible ratelimiting step for ArA biosynthesis. The presence of several putative c is-acting regulatory elements in this identified promoter sheds new light on the regulation mechanism research of Δ6 FAD transcription for the ArA production in M. incisa in changing environmental factors.
基金supported by China’s National Special Research Fund for Non-Profit Marine Sector (No. 2013418027)Marine Biology Program of Shanghai Leading Academic Disipline (No. J50701)+1 种基金Marine Special Fund of Shanghai Committee of Science and Technology (No. 11dz1205000)a Doctoral Research Fund from Shanghai Ocean University (No. A2-0302-14-300084)
文摘Invasive species generally possess unique characteristics that allow them to survive the invasion process in order to establish and spread in new habitats. Successful invaders must resist both physical and physiological stresses associated with the changing environment. A common littoral barnacle, Chthamalus challengeri Hoek, 1883(Crustacea, Cirripedia), which is native to Japan, South Korea and northern China, has become established in the high-littoral zone adjacent to Yangshan Port, Shanghai, China. A comparison of the morphology of Chthamlus species from Zhoushan archipelago with previous description indicates the occurrence of C. challengeri. The new immigrant becomes a dominant species in certain high-intertidal habitats of the adjacent area to of Yangshan Port. C. challengeri was found in part of sampling sites in Zhoushan in 2010; however, it dispersed to all the eleven sampling sites in 2012. Densities of C.challengeri had increased over 10 times in the last 2 years, with the highest mean value reaching 39533 ± 6243 ind. m-2 in the new habitat. The specific ratios of both operculum area(Sa) to base area(SA) and average height of parietal plates(H) to length of base(L) revealed that C. challengeri displays morphological changes to resist stronger currents in the new habitats for invasion.
基金Supported by the National Natural Science Foundation of China(Nos.41376136,31201992)the Double First-Class Discipline of Fisheries Science in China。
文摘It has been reported that there was a linkage of 5S rRNA gene to 18S-5.8S-25S rRNA gene in a few of species in Ochrophyta.In regard to the usual two positions of linked 5S rDNA to the 3′end of 25S rDNA,two pairs of primers were designed for amplification to verify this linkage of two genes in a kelp cultivar of Saccharina japonica,one of species in Ochrophyta.This result supplemented the previous report that 5S rDNA was unlinked to 25S rDNA in this kelp.In order to simultaneously visualize this unlinkage of two genes,dual-color fluorescence in situ hybridization(FISH)technique was applied to the cytogenetics of S.japonica.Dual-color FISH images showed that two and four hybridization signals were present in the kelp gametophyte and sporophyte,respectively,metaphase nuclei hybridized simultaneously with the labeled probes of 18S rDNA and 5S rDNA.Both haploid and diploid karyotypes in decreasing length of chromosomes showed that 18S-5.8S-25S rDNA was localized at the interstitial region of Chromosome 23,whereas 5S rDNA resided at the sub-telomeric region of Chromosome 27.These karyotypes suggested that the kelp nuclear genome had only one locus of each rRNA gene,and their loci on different chromosomes indicated the physical unlinkage of 5S rDNA to 18S-5.8S-25S rDNA in this kelp.Therefore,dual-color FISH seems to be a powerful technique for the discrimination and pairing of chromosomes featured in both small size and nearly identical shape in S.japonica.
基金funded by the National Natural Science Foundation of China(Grant number:32030112)National Key R&D Program of China(Grant number:2018YFD0900302).
文摘Interleukin-21(IL-21)is produced mainly by activated CD4^(+)T cells and promotes the proliferation,survival and differentiation of immune cells.This study characterised a high affinity binding receptor of IL-21(IL-21 Rα)from grass carp(Ctenopharyngodon idella,Ci)and analyzed the expression of CiIL-21 and CiIL-21 Rα.It has been shown that they were modulated by PAMPs and during infection with bacterial and viral pathogens.Recombinant CiIL-21 protein produced in bacteria was active in inducing the expression of genes involved in Th responses.A monoclonal CiIL-21 antibody was generated against the rCiIL-21 and showed good reactive specificity with the rCiIL-21 proteins expressed in E.coli cells,and HEK293 T cells and the native protein from PHA-stimulated kidney leukocytes.In addition,the IL-21-producing cells detected by the CiIL-21 monoclonal antibody were found to be increased to in the kidney,spleen and hindgut after infection with Flavobacterium columnare.Our data indicate that IL-21 is involved in the regulation of immune response to bacterial and viral infection.
基金supported by the National Key Research and Development Project(2018YFD0900301)the National Natural Science Foundation of China(31802325).
文摘Miiuy croaker,Miichthys miiuy is an ecologically important teleost species which is widely distributed in southeast coast of China.In this study,we present a chromosomal-scale genome assembly of the miiuy croaker which is an important Sciaenidae fish and economical species.We adopted Oxford Nanopore and Hi-C sequencing techniques to achieve an assembly with high accuracy and completeness.The investigation of genome characteristic and functional features may provide insights into the study of phylogenetic diversity of miiuy croaker.This study can also be beneficial to improve molecular assisted breeding techniques.Moreover,it can be a great resource to better conduct further study of other sciaenids.
基金funded by the National Natural Science Foundation of China(Grant number:32030112)National Key R&D Program of China(Grant number:2018YFD0900302).
文摘CD4 T helper cells are an important group of cells in the immune system of vertebrates and express CD4 receptor on the cell surface.In mammals,the CD4 receptor is encoded by a single copy gene,whilst in fish,two copies of cd4 genes,namely cd4-1 and cd4-2,are found.In this study,the ectodomains of grass carp(Ctenopharyngodon idella,Ci)CD4-1 and CD4-2 were expressed in the E.coli cells and used to generate monoclonal antibodies in mice.Western blotting,confocal microscopy and flow cytometry were performed to characterize the monoclonal antibodies.It has been shown that the CiCD4-1 and CiCD4-2 monoclonal antibodies had good specificity to react with the recombinant ectodomains of CiCD4-1 and CiCD4-2 expressed in the CHO-S cells and the native CD4 molecules of grass carp.The CiCD4-1 monoclonal antibody did not recognize CiCD4-2 and verse versa.In addition,the CD4-1 and CD4-2 monoclonal antibodies specifically recognized the CD4-1 and CD4-2 receptors expressed in the HEK293 cells and native molecules of fish cells.Further,the percentages of lymphocytes in immune tissues of healthy fish were analyzed by flow cytometry.It was found that 17.6%of lymphocytes were CD4-1^(+)cells and 22.5%were CD4-2^(+)in the head kidney.In the spleen,13.1%of lymphocytes were CD4-1^(+)and 18.6%were CD4-2^(+)while 7.3%of blood lymphocytes were CD4-1^(+)cells and 8.8%were CD4-2^(+)cells.The availability of the CD4-1 and CD4-2 monoclonal antibodies provides antibody-based tools for further elucidation of the functions of CD4 T cells in grass carp.
基金Supported by the Aquaculture and Fisheries Collaborative Research Support Program of USAID (No. 1366)the Shanghai Rising-Star Program (No. 08QA1405900)+1 种基金the Innovation Program of Shanghai Municipal Education Commission (No. 09YZ277)the Shanghai Leading Academic Discipline Project (No. Y1101)
文摘Cyanobacterial blooms in eutrophic freshwater systems are a worldwide problem, creating adverse effects for many aquatic organisms by producing toxic microcystins and deteriorating water quality. In this study, microcystins (MCs) in Microcystis aeruginosa, and Daphnia magna exposed to M. aeruginosa, were analyzed by HPLC-MS, and the effects of M. aeruginosa on D. magna were investigated. When D. magna was exposed to M. aeruginosa for more than 2 h, Microcystin-LR (MC-LR) was detected. When exposed to 1.5 × 106, 3 × 106, 0.75 × 107, and 1.5 × 107 cell/mL of M. aeruginosa for 96 h, average survival of D. magna for treatments were 23.33%, 33.33%, 13.33%, 16.67%, respectively, which were significantly lower than the average 100% survival in the control group (P < 0.05). The adverse effects of M. aeruginosa on body length, time for the first brood, brood numbers, gross fecundity, lifespan, and population growth of D. magna were density-dependent. These results suggest that the occurrence of M. aeruginosa blooms could strongly inhibit the population growth of D. magna through depression of survival, individual growth and gross fecundity. In the most serious situations, M. aeruginosa blooms could undermine the food web by eliminating filter-feeding zooplankton, which would destroy the ecological balance of aquaculture water bodies.
基金"863" Hi-tech Research and Development Program of China under contract No. 2006 AA10 A410Shanghai Leading Academic Discipline Project under contract No. Y1101Dr. Start-up Fund under contract No. A-2400-09-0144
文摘The nucleotide sequence of the mitochondrial genome of the solecurtidae Bivalvia mollusca Sinonovacula constricta(GenBank accession number EU880278) has been determined and is reported here.We determined the complete mitochondrial genome sequence using long-PCR and Shot Gun Sequencing.Contained within the 17 225 base pairs(bp) are the two ribosomal RNA genes and 12 protein coding genes typical of metazoan mitochondrial genomes.The S.constricta mitochondrial DNA(mtDNA) did not contain a gene for atp8,similar to the mtDNA of Crassostrea virginica,Crassostrea giga and Mytilus edulis.The S.constricta mtDNA is 67.0% A+T(A 25.9%,C 10.5%,G 22.5%,and T 41.1%).This value is higher than that for many invertebrate mitochondrial genomes.Only 19 putative tRNA genes are present in S.constricta and 27 noncoding regions,of which two are large in size.The trnE and trnW genes as well as a second trnS were absent in S.constricta.The gene arrangement of S.constricta is different from the other Bivalvia genomes.
基金supported by the China Postdoctoral Science Fundation(No.2019M651472)the National Natural Science Foundation of China(No.31672642)the Key Laboratory of Mariculture of Ministry of Education,Ocean University of China(No.KLM2018009)。
文摘Pituitary adenylate cyclase activating polypeptide(PACAP)and growth hormone-releasing hormone(GHRH)play important roles in the GH/IGF growth axis in fishes.To determine whether epigenetic change is involved in the regulation of pacap and ghrh responses to low salinity stress in Cynoglossus semilaevis,the correlation between growth traits,DNA methylation status and gene expression level in low salinity(15,S15)and optimal salinity(30,S30)at day 7(D7)and day 60(D60)were analyzed.Results showed that exposure to low salinity for 60 days attenuated C.semilaevis growth rate.Under low salinity,DNA methylation level of pacap promoter increased in females and decreased in males at day 7,but inverted at day 60.Additionally,pacap expression was up-regulated in both males and females.The pacap promoter methylation level was higher and expression level was lower in female than in male.The results suggest that pacap promoter methylation level is negatively correlated to mRNA level and positively correlated to body weight,while gene expression level is negatively related with body weight.With the decrease of salinity,DNA methylation level of ghrh promoter and exon1,as well as its gene expression displayed minor changes.Overall,pacap gene seems to play an important role in fish growth,contributing to female growth superiority,while ghrh gene seems not pertinent under salinity stress.The results indicate that low salinity potentially affects fish growth through regulating DNA methylation in pacap promoter.This study expands the understanding of the molecular mechanism of how salinity modulates fish growth from the epigenetic perspective.
基金the National Natural Science Foundation of China(No.31872587)the CAS Key Laboratory of Coastal Environmental Processes and Ecological Remediationthe YICCAS Grant(No.2020 KFJJ11)。
文摘The population of tri-spine horseshoe crabs(HSCs)Tachypleus tridentatus has sharply decreased in recent decades.Domestication and manual releasing are now used to recover it.In order to accelerate the growth of T.tridentatus in indoor environment,in this study,gut microbiota of 2nd instar T.tridentatus was supposed to be improved by putting probiotics in water environment and thus promoted the health and growth of the host.Using 16S rRNA sequencing of gut samples,we examined the intestinal microbiome of juvenile HSCs and studied the effect of probiotics.Results showed that there was no significant difference between groups fed with high and low concentrations(1×10^(7),1×10^(12)CFU mL^(−1))of Bacillus subtilis and high concentration(1×10^(12)CFU mL^(−1))of Lactobacillus,and no significant difference compared with the control group,but the diversity of Lactobacillus group with low concentration(1×10^(7)CFU mL^(−1))was significantly higher than those of other groups.Microbial-mediated functions predicted by PICRUSt showed that‘Cellular Processes’,‘Metabolism’and‘Organismal Systems’pathways significantly increased in groups of Bacillus subtilis and Lactobacillus group with low concentration.The results showed that two types of probiotics with different concentrations have other effects on the structure,richness and function of intestinal microbes.By searching the literature and conducting pre-experiments,we forecasted the short time of breeding under the probiotic environment may not have much effect.The present study for the first time investigated the impact of probiotics on gut microbiota of juvenile HSCs and found them may benefit for the growth and health of HSCs.
基金funded by the National Natural Science Foundation of China(Grant No.32030112 and U21A20268).
文摘A cell line,termed ZFIN,was established from the caudal fin of zebrafish and was shown to be susceptible to spring viremia of carp virus(SVCV).The ZFIN cells are epithelial like cells and have a moderate plasmid transfection efficiency of 13.9%.Using an RNA-seq approach,differentially expressed genes(DEGs)regulated by SVCV were identified.Infection of SVCV gave rise to 3931 DEGs and up-regulated DEGs were mostly enriched into the biological regulation and cellular processes,among which pathways for the type I interferon signaling and the response to exogenous dsRNA were the top two GO terms.Several KEGG signaling pathways including TLR signaling pathway,RLR receptor signaling pathway,cytosolic DNA-sensing pathway,NLR signaling pathway,cytokine-cytokine receptor interaction and ferroptosis were significantly enriched.Antiviral genes including ifnφ1,isg15 and mx were significantly up-regulated.In addition,key DEGs involved in autophagy were identified.The results indicate that the ZFIN cell line provides a useful in vitro tool for study on the gene functions and cellular responses to viral infection in fish.
基金from Shanghai Agriculture Applied Technology Development Program,China(Grant No.G20190102)National Natural Science Foundation of China(Grant No.81770165 to BH)Shanghai Municipal Education Commission(Project for Gaofeng Discipline of Fishery).
文摘Lysine-specific demethylase 4A(KDM4A)catalyzes demethylation of histone lysine residues,which regulates chromatin state and transcription.In drosophila and mice,KDM4A plays an important role in multiple biological processes including development,aging,metabolism,and immunity,however the functions of KDM4A in fish are still unclear.There are two copies of the kdm4a gene in zebrafish,namely kdm4aa and kdm4ab,kdm4aa was edited using CRISPR/Cas9 technology in the present study,then homozygous kdm4aa mutants(kdm4aa^(-/-))were obtained,and loss of kdm4aa was confirmed by sequencing and increased H3K9me3.Whole-mount in situ hybridization showed that kdm4aa is widely expressed during the embryonic development of zebrafish.Compared with WT zebrafish,kdm4aa^(-/-)zebrafish showed no significant difference in gamete formation and fertilization,but the survival rate of kdm4aa^(-/-)embryos dramatically reduced to 21%at 26 hpf.Further observation showed that about 80%of survived kdm4aa^(-/-)zebrafish experienced disruption in stripe formation,and 10%of survived kdm4aa^(-/-)zebrafish underwent vertebral malformation.Alizarin red S staining demonstrated the abnormal spinal development in kdm4aa^(-/-)zebrafish.These results indicated that kdm4aa is required for normal embryonic development of zebrafish,loss of kdm4aa function leads to decreased survival during the early stages of zebrafish development and morphological variation in adult zebrafish.
基金supported by the National Key Research and Development Project (2018YFD0900503)National Natural Science Foundation of China (31802325,31822057)。
文摘Noncoding RNA(ncRNA)is an important regulatory factor that plays a major role in innate immunity.However,most studies on ncRNA have focused on mammals,resulting in a knowledge gap on nc RNA in lower vertebrates such as teleost fish.In this study,we identified a new long noncoding RNA(lncRNA),termed TAK1-related lnc RNA(TARL),which can play a positive role in the antibacterial immunity of Miichthys miiuy to Vibrio anguillarum and V.harveyi.We also found a novel micro RNA miR-2188-3p that could target TAK1 and inhibit the host antibacterial response and promote bacterial escape.We further found that the antibacterial effect inhibited by miR-2188-3p could be reversed with TARL.Moreover,V.anguillarum and V.harveyi are the two most susceptible Gram-negative pathogens of aquaculture fish,and the economic losses caused by these two bacteria are immeasurable every year.This study is the first to report on the ability of lnc RNA to prevent the escape of V.anguillarum and V.harveyi in fish through the competing endogenous RNA(ceRNA)mechanism.Our results not only elucidate the ce RNA mechanism of the lncRNA in antibacterial immune responses but also provide new insights into the impact of lnc RNA on host immunity and bacterial escape.
基金the National Natural Science Foundation of China[31872546].
文摘Activin A receptor,type II(Acvr2)is a member of the transforming growth factor beta receptor family and can function as a negative regulator of skeletal muscle mass.Acvr2 plays an important role in regulating muscle development that can inhibit skeletal muscle growth in mice.However,there is very little research reported on the function of acvr2 in muscle development of teleost.In this study,we analyzed the effect of acvr2aa and acvr2ba on muscle development in zebrafish.Growth rates of WT and acvr2a^(-/-b-/-)were measured from juvenile stage to adult stage.In addition,effects of acvr2 on skeletal muscle were tested in histological,protein and molecular levels.As a result,acvr2a^(-/-b-/-)exhibited a wider body trunk than WT and showed a significant increase in body weight and width from two months old.Histological analysis of skeletal muscle indicated that the size of muscle fiber in acvr2a^(-/-b-/-)(female:1809±123μm^(2);male:2261±130μm^(2))was larger than that in WT(709.8±49μm^(2);815±53μm^(2)).In addition,western blot of fast MyLc protein showed the protein synthesis of acvr2a^(-/-b-/-)are increased.Besides,Histological analysis of heart showed the ventricle area is aslo increased in acvr2a^(-/-b-/-).Our results demonstrated acvr2 attends the development of muscle fiber and will cause muscle hypertrophy when they were knocked out in zebrafish.In conclusion,acvr2 in zebrafish can control the development of muscle fibers during posthatch growth.
基金supported by the National Key Research and Development Program of China(2018YFD0900606)National Natural Science Foundation of China(31072208)+2 种基金Major Science and Technology Specific Program of Zhejiang Province,China(2016C02055-6)Science and Technology Planning Project of Jiangsu Province,China(BE2018335)Open Program of Key Laboratory of Cultivation and High-value of Marine Organisms in Fujian Province,China(2017fjscq02).
文摘As an important seedling source,monospores closely associate with yields in nori farming.However,the molecular mechanism underlying differences in monospore production for different strains remains unknown.Comparative transcriptome analysis was performed to examine gene expression differences between the spore abundant wild-type strain(WT)and spore deficient mutant(Y1)of Pyropia chauhanii.The WT strain that produces monospores in abundance exhibited more differentially expressed genes(DEGs)in both number and higher fold-changes than the Y1 strain incapable of producing monospores,indicating that the specific regulation of genes is involved in monospore production.Three lists of DEGs were obtained between the two strains using intersection and displayed in Venn diagram:one expressed only in WT strain,another expressed only in Y1 strain,and the third shared in both strains.DEGs annotated as homologous genes of Arabidopsis thaliana in these 3 lists were curated for online functional enrichment analysis on Metascape website.Gene regulatory networks of WT were functionally enriched in the processing,proteolysis,and transport of proteins,especially within the small GTPase protein family,which might be account for the monospore production ability,whereas Y1 were functionally enriched in the metabolism of essential substance and utilization of indispensable energy,which might be account for the rapid growth of blades.We found the differentially enriched gene regulatory networks between strains might be the intrinsic mechanisms of the different monospore production traits.These findings provide novel insights into the genes and regulatory networks associated with monospore production abilities,which are essential for developing accurate breeding technologies for optimal release of monospores and increase of total nori production.
