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1.
Increasingly arisen environmental constraints may contribute to heritable phenotypic variation including methylation changes,which can help the animals with development,growth and survival.In this study,we assessed the DNA methylation levels in three tissues(gonad,kidney and gill) of half smooth tongue sole under the salinity stress.The methylation-sensitive amplification polymorphism(MSAP) technique was applied to illustrate the regulation of epigenetic mechanism in environmental stimuli.Fish were subjected to 15 salinity treatment for 7 and 60 days,respectively.A total of 11259 fragments were amplified with 8 pairs of selective primers.The levels of methylated DNA in different tissues of females and males without salinity stress were analyzed,which were 32.76% and 47.32% in gonad;38.13% and 37.69% in kidney;37.58% and 34.96% in gill,respectively.In addition,the significant difference was observed in gonad between females and males,indicating that discrepant regulation in gonadal development and differentiation may involve sex-related genes.Further analysis showed that total and hemi-methylation were significantly decreased under 15 salinity for 7 days,probably resulting in up-regulating salt-tolerance genes expression to adjust salt changing.With the adjustment for 60 days,total and hemi-methylation prominently went back to its normal levels to obtain equilibrium.Particularly,full methylation levels were steady along with salinity stress to maintain the stability of gene expression.Additionally,the data showed that gonads in females and gills in males were superior in adaptability.As a result,DNA methylation regulates tissue-specific epiloci,and may respond to salinity stress by regulating gene expression to maintain animal survival and activity.  相似文献   

2.
Increasingly arisen environmental constraints may contribute to heritable phenotypic variation including methylation changes, which can help the animals with development, growth and survival. In this study, we assessed the DNA methylation levels in three tissues (gonad, kidney and gill) of half smooth tongue sole under the salinity stress. The methylation-sensitive amplification polymorphism (MSAP) technique was applied to illustrate the regulation of epigenetic mechanism in environmental stimuli. Fish were subjected to 15 salinity treatment for 7 and 60 days, respectively. A total of 11259 fragments were amplified with 8 pairs of selective primers. The levels of methylated DNA in different tissues of females and males without salinity stress were analyzed, which were 32.76% and 47.32% in gonad; 38.13% and 37.69% in kidney; 37.58% and 34.96% in gill, respectively. In addition, the significant difference was observed in gonad between females and males, indicating that discrepant regulation in gonadal development and differentiation may involve sex-related genes. Further analysis showed that total and hemi-methylation were significantly decreased under 15 salinity for 7 days, probably resulting in up-regulating salt-tolerance genes expression to adjust salt changing. With the adjustment for 60 days, total and hemi-methylation prominently went back to its normal levels to obtain equilibrium. Particularly, full methylation levels were steady along with salinity stress to maintain the stability of gene expression. Additionally, the data showed that gonads in females and gills in males were superior in adaptability. As a result, DNA methylation regulates tissue- specific epiloci, and may respond to salinity stress by regulating gene expression to maintain animal survival and activity.  相似文献   

3.
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.  相似文献   

4.
In this study, the methylation-sensitive amplification polymorphism(MSAP) was used to compare the genomic DNA methylation level of muscle, gill and hepatopancreas of Portunus trituberculatus subjected to salinity 12 for 30 days to illustrate the epigenetic mechanism of osmoregulation. Thirty primers were used to analyze the difference of methylation level of different tissues. The results showed that the baseline methylation level of muscle, hepatopancreas and gill was 47.31%, 22.94% and 17.69%, respectively. After exposed to low salinity stress, the methylation epiloci changed in the three tissues. Both demethylation and methylation processes occurred under low salinity stress. The methylation ratio decreased in muscle and gill but increased in hepatopancreas. These results indicated that DNA methylation is tissue-specific when P. trituberculatus responds to low salinity.  相似文献   

5.
Kong  Ning  Liu  Xiao  Li  Junyuan  Mu  Wendan  Lian  Jianwu  Xue  Yanjie  Li  Qi 《中国海洋湖沼学报》2017,35(5):1248-1258
Temperature and salinity are two of the most potent abiotic factors infl uencing marine mollusks.In this study,we investigated the individual and combined ef fects of temperature and salinity on the survival and growth of juvenile Pacifi c abalone,Haliotis discus hannai Ino,and also examined the DNA methylation alteration that may underpin the phenotypic variation of abalone exposed to dif ferent rearing conditions.The single-factor data showed that the suitable ranges of temperature and salinity were 16–28°C at a constant salinity of 32,and 24–40 at a constant temperature of 20°C,respectively.The two-factor data indicated that both survival and growth were signifi cantly af fected by temperature,salinity and their interaction.The optimal temperature-salinity combination for juveniles was 23–25°C and 30–36.To explore environment-induced DNA methylation alteration,the methylation-sensitive amplifi ed polymorphism(MSAP)technique was used to analyze the genomic methylation profi les of abalone reared in optimal and adverse conditions.Neither temperature nor salinity induced evident changes in the global methylation level,but 67 and 63 dif ferentially methylated loci were identifi ed in temperature and salinity treatments,respectively.The between-group eigen analysis also showed that both temperature and salinity could induce epigenetic dif ferentiation in H.discus hannai Ino.The results of our study provide optimal rearing conditions for juvenile H.discus hannai Ino,and represent the fi rst step toward revealing the epigenetic regulatory mechanism of abalone in response to thermal and salt stresses.  相似文献   

6.
Galectins,a family of ?-galactoside-binding proteins,participate in both innate immunity and adaptive immunity.This study identified one novel galectin-related protein from half-smooth tongue sole Cynoglossus semilaevis,which was designated as Cs GRP.The full-length c DNA of its encoding gene was 785 bp in length with a 528 bp open reading frame encoding a putative protein of 176 amino acids.The deduced Cs GRP contained a putative 131-aa galactoside-binding lectin domain and 3 critical residues responsible for carbohydrate binding(R~(93),W~(109) and R~(114)).Genomic structural analysis revealed that Cs GRP consisted of five exons and four introns.Cs GRP showed 68% similarity with Poecilia latipinna GRP and 67% similarity with Stegastes partitus GRP.Cs GRP showed the highest expression level in liver,although its expression was detected in all tested tissues.When challenged with Vibrio harveyi,the expression of Cs GRP was significantly down-regulated in liver(P 0.05).In addition,we found that in spleen and kidney of C.semilaevis,the Cp G island of Cs GRP showed significantly higher(P 0.05) methylation level in disease-resistant family of C.semilaevis(DR-Cs) than in disease-susceptible ones(DS-Cs).Our results suggested that Cs GRP may play important roles in the immune response of C.semilaevis.Moreover,DNA methylation patterns provided valuable data for understanding the relationship between epigenetic regulation and immunity,which would assist the animal genetic research and improve the animal breeding in the future.  相似文献   

7.
The sea cucumber, Apostichopus japonicus, undergoes aestivation to improve survival during periods of high-temperature. During aestivation, the metabolic rate is depressed to reduce the consumption of reserved energy. We evaluated the role of epigenetic modification on global gene silencing during metabolic rate depression in the sea cucumber. We compared the expression of epigenetic modifiers in active and aestivating sea cucumbers. The expression of three genes involved in DNA methylation and chromatin remodeling (DNA (cytosine-5)-methyltransferase 1, Methyl-CpG-binding domain protein 2), and Chromodomain-helicase-DNA-binding protein 5) was significantly higher during aestivation (Days 20 and 40). Similarly, we observed an increase in the expression of genes involved in histone acetylation (Histone deacetylase 3) and Histone-binding protein RBBP4) during the early (Days 5 and 10) and late phases (Days 20 and 40) of aestivation. There was no change in the expression of KAT2B, a histone acetyltransferase. However, the expression of histone methylation associated modifiers (Histone-arginine methyltransferase CARMER and Histone-lysine N-methyltransferase MLL5) was significantly higher after 5 d in the aestivating group. The results suggest that the expression of epigenetic modifiers involved in DNA methylation, chromatin remodeling, histone acetylation, and histone methylation is upregulated during aestivation. We hypothesize that these changes regulate global gene silencing during aestivation in A. japonicus.  相似文献   

8.
Half-smooth tongue sole (Cynoglossus semilaevis) is a promising species for aquaculture in China.The wild population of C. semilaevis is under threat from environmental factors. Microsatellite markers are very suitable for assessing genetic diversity. Four microsatellite-enriched libraries of half smooth tongue sole (Cynoglossus semilaevis) were constructed,from which 57 polymorphic microsatellites were isolated and characterized.The polymorphism of these microsatellites was assessed by genotyping in 30 ind...  相似文献   

9.
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10.
Jiao  Shuang  Tan  Xungang  You  Feng  Zhang  Shujing  Pang  Qiuxiang 《中国海洋湖沼学报》2023,41(1):280-289

Insulin-like growth factors (IGFs) are key regulators of development and growth. Here, we characterized the igf2 gene from olive flounder (Paralichthys olivaceus) and determined its temporal and spatial expression. We set up an in-vitro protein expression system in eukaryotic human embryonic kidney (HEK293T) cells and explored its effects on cell proliferation. The flounder igf2 cDNA contained a 648-bp open reading frame (ORF) encoding a protein of 215 amino acids (aa), which spanned the complete signal peptide (47 aa), mature peptide (70 aa), and E domain (98 aa). In adult flounder, igf2 mRNA was detected in all selected tissues. In early development, igf2 mRNA was detected throughout development from unfertilized eggs to hatching-stage embryos. In-situ hybridization analysis indicated that igf2 mRNA was specially expressed in the brain region, floor plate, hypochord, otic vesicle, and pectoral fin during embryogenesis. Western blotting analysis indicated that the soluble recombinant flounder IGF2 protein was successfully produced through eukaryotic expression in HEK293T cells. In addition, the recombinant IGF2 protein significantly promoted the proliferation of human cervical carcinoma (HeLa) and HEK293T cells. These results provide new information about the structural and functional conservation, expression patterns, and biological activity of the igf2 in teleosts.

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11.
以勒氏笛鲷(Lutjanus russellii)基因组DNA为模板,采用同源克隆的方法,获得2 887 bp的肌肉生长抑制素(myostatin,MSTN)基因组序列,该MSTN序列具有3个外显子和2个内含子,包括101 bp的5′-UTR、385bp的外显子1、354 bp的内含子1、370 bp的外显子2、761 bp的内含子2、381 bp的外显子3和1 932 bp的3′-UTR。整个开放阅读框编码了378个氨基酸,前面的22个氨基酸为信号肽,具有9个保守的半胱氨基酸及一个RVRR的蛋白酶解加工位点。氨基酸序列分析发现,该基因编码的蛋白质与其他鱼类的I型同源性较高,与其他鱼类的2型MSTN同源性较低,且与鲈形目的同源性最高,与鲤形目的同源性较低,与人、鼠和鸡的同源性最低。采用邻接法(Neighbor-Joining)构建的MSTN的系统发育树表明,勒氏笛鲷MSTN与鲈形目鱼类的狼鲈属亲缘关系较近,且与鱼类的MSTN-1聚为1支。这表明该基因属于Ⅰ型MSTN基因。  相似文献   

12.
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13.
In order to depict the distribution of diatom fossils in surface sediments and to establish a reliable reference data for further paleoenvironmental study in the Changjiang(Yangtze) River estuary and its adjacent waters, the diatom fossils from 34 surface sediment samples and their relationship with environmental variables were analyzed by principal component analysis and redundancy correspondence analysis. The diversity and abundance of diatom fossils were analyzed. Some annual average parameters of the overlying water(salinity, temperature, turbidity, dissolved oxygen, depth, dissolved inorganic nitrogen,dissolved inorganic phosphate and dissolved inorganic silicate) were measured at each sampling site. A total of 113 diatom taxa and one silicoflagellate species were identified in the investigation area. Diatom fossils were better preserved in fine sediments. The absolute abundance of diatom fossils did not significantly dif fer between inshore and of fshore areas, the species diversity decreased from inshore to of fshore. This may be because high nutrients and low salinity promoted the growth of more brackish species in coastal waters. The diatom taxa were divided into three groups, on the basis of their response and indication to environmental changes. For example, Actinocyclus ehrenbergii and Cyclotella stylorum were dominant in coastal waters(Group 1 and Group 3) with high nutrients and low salinity; the relative abundances of Paralia sulcata and Podosira stelliger were significantly higher in of fshore sites(Group 2, average 39.5%),which were characterized by high salinity and deep water. Four environmental variables(salinity, dissolved inorganic nitrogen, temperature and water depth) explained the composition and distribution of diatom taxa independently( P 0.05), this finding can be applied in further paleoenvironmental reconstruction research in this area.  相似文献   

14.
As a specific gene of fish,cytochrome P450c17-Ⅱ(CYP17-Ⅱ) gene plays a key role in the growth,development and reproduction level of fish.In this study,the single-stranded conformational polymorphism(SSCP) technique was used to characterize polymorphisms within the coding region of CYP17-Ⅱ gene in a population of 75 male Japanese flounder(Paralichthys olivaceus).Three single nucleotide polymorphisms(SNPs) were identified in CYP17-Ⅱ gene of Japanese flounder.They were c.G594A(p.G188R),c.G939A and c.G1502A(p.G490D).SNP1(c.G594A),located in exon 4 of CYP17-Ⅱ gene,was significantly associated with gonadosomatic index(GSI).Individuals with genotype GG of SNP1 had significantly lower GSI(P < 0.05) than those with geno-type AA or AG.SNP2(c.G939A) located at the CpG island of CYP17-Ⅱ gene.The mutation changed the methylation of exon 6.Indi-viduals with genotype AA of SNP2 had significantly lower serum testosterone(T) level and hepatosomatic index(HSI) compared to those with genotype GG.The results suggested that SNP2 could influence the reproductive endocrine of male Japanese flounder.How-ever,the SNP3(c.G1502A) located in exon 9 did not affect the four measured reproductive traits.This study showed that CYP17-Ⅱ gene could be a potentially useful candidate gene for the research of genetic breeding and physiological aspects of Japanese flounder.  相似文献   

15.
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16.
Gracilariopsis lemaneiformis is an economically important agarophyte, which contains high quality gel and shows a high growth rate. Wild population of G. lemaneiformis displayed resident divergence, though with a low genetic diversity as was revealed by amplified fragment length polymorphism (AFLP) and simple sequence repeat (SSR) analyses. In addition, different strains of G. lemaneiformis are diverse in morphology. The highly inconsistence between genetic background and physiological characteristics recommends strongly to the regulation at epigenetic level. In this study, the DNA methylation change in G. lemaneiformis among different generation branches and under different temperature stresses was assessed using methylation sensitive amplified polymorphism (MSAP) technique. It was shown that DNA methylation level among different generation branches was diverse. The full and total methylated DNA level was the lowest in the second generation branch and the highest in the third generation. The total methylation level was 61.11%, 60.88% and 64.12% at 15°C, 22°C and 26°C, respectively. Compared with the control group (22°C), the fully methylated and totally methylated ratios were increased in both experiment groups (15°C and 26°C). All of the cytosine methylation/demethylation transform (CMDT) was further analyzed. High temperature treatment could induce more CMDT than low temperature treatment did.  相似文献   

17.
A complementary DNA (cDNA) library was constructed from half-smooth tongue sole spleen. A long-read expressed sequence tag (EST) database was generated, containing 3100 cDNA clones, of which 220 clones were fully sequenced. A total of 1060 non-redundant simple sequence repeats (SSRs) were obtained from the cDNA library. An average of 5 kb sequence generates 1 SSR in the half-smooth tongue sole spleen cDNA library. The proportion of the SSR unit size was different in the cDNA library. The monomeric repeats (51.4%) are the most abundant class of SSR in the dataset. The dimeric, trimeric, tetrameric and hexameric repeats are represented in decreasing proportions of 27.2%, 16.0%, 2.8% and 1.9%, respectively. The frequency of pentameric repeats was observed the least (only 0.7%). Most of the monomeric and dimeric repeats are distributed in 3′ and 5′ un-translation region. If translation regions are considered merely, trimeric repeats are the highest, accounting for 57% of the total microsatellites.  相似文献   

18.
A complementary DNA (cDNA) library was constructed from half-smooth tongue sole spleen. A long-read expressed sequence tag (EST) database was generated, containing 3100 cDNA clones, of which 220 clones were fully sequenced. A total of 1060 non-redundant simple sequence repeats (SSRs) were obtained from the cDNA library. An average of 5 kb sequence generates 1 SSR in the half-smooth tongue sole spleen cDNA library. The proportion of the SSR unit size was different in the cDNA library. The monomeric repeats (51.4%) are the most abundant class of SSR in the dataset. The dimeric, trimeric, tetrameric and hexameric re- peats are represented in decreasing proportions of 27.2%, 16.0%, 2.8% and 1.9%, respectively. The frequency of pentameric repeats was observed the least (only 0.7%). Most of the monomeric and dimeric repeats are distributed in 3' and 5' un-translation region. If translation regions are considered merely, trimeric repeats are the highest, accounting for 57% of the total microsatellites.  相似文献   

19.
The Chinese tongue sole(Cynoglossus semilaevis) is widely cultured in the coastal region of East Asia and has excellent economic value. However, the high albino rate of the breeding population has caused a significant loss to the aquaculture industry. To study the molecular mechanism of albinism, the present study used an albino Chinese tongue sole family to construct three simple sequence repeat(SSR) linkage groups, and draft a preliminary linkage map related to albinism. After albinism-related loci mapping, 18 albinism-related loci were detected under two models(containing 2407 genes) compared to the Chinese tongue sole genome. One of these loci, the tyrosinase related protein(tyrp2), which has been reported previously as an important gene regulating both eumelanin and phaeomelanin levels, was indicated to be the possible cause of albinism. Thirty-five Gene Ontology(GO) terms and 14 Kyoto Encyclopedia of Genes and Genome pathways were annotated via bioinformatic analyses. One GO term with protein tyrosine kinase activity, which contained 10 genes, was previously suggested to affect fish albinism. These results establish a foundation for further in-depth study of albinism in Chinese tongue sole.  相似文献   

20.
Studies were conducted to determine the cause of the acute mortality of half-smooth tongue sole Cynoglossus semilaevis Günther juveniles in a fish farm in Jimo, Shandong Province, China, in June 2006. Gross signs of the diseased tongue sole included several petechiae and ecchymoses on the body and fin necrosis and hemorrhagic lesion at the base of the fin. Bacteria were isolated from kidney, liver and hemorrhagic lesions of the diseased tongue sole. Among14 strains, SJ060621 was proved to be highly virulent to juvenile tongue sole with LD50 value of 1.0×105 colony forming units (CFU)mL-1, while the remaining 13 were avirulent. Among the 16 antibiotics tested, SJ060621 was sensitive to gentamicin and nitrofurantoin. It was identified as Listonella anguillarum with conventional plate and tube tests in combination with API 20E analysis. 16S rRNA gene and partial HSP60 gene sequenceing analysis revealed that the strain was highly homologous with L. anguillarum. Examination of the infected musculature by electron microscopy indicated numerous bacteria and lots of macrophages containing phagocytosed bacteria. Histopathological investigations revealed severe necrotic degenerative changes in the infected organs. Indirect immunofluorescence assay (IFA) was employed to detect the location of occurrence of bacteria, and bacteria were found in aggregations in the inflammatory areas in musculature.  相似文献   

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