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【目的】探索波吉卵囊藻胞外滤液对铜绿微囊藻的抑制机理。【方法】采用Illumina平台,对使用BG11培养基(对照组)和波吉卵囊藻胞外滤液(实验组)培养的铜绿微囊藻(Microcystis aeruginosa)进行转录组测序,筛选差异表达基因并进行通路富集分析,并对部分差异表达基因进行qRT-PCR验证。【结果】与对照组相比,波吉卵囊藻胞外滤液处理后的铜绿微囊藻组中筛选1483个表达差异显著的基因,其中上调表达基因788个,下调表达基因695个。GO功能分析将差异基因划分为35个子类别,包括催化活性、结合、代谢过程、细胞过程、膜、膜部分等。KEGG通路分析中,所有差异基因聚集在108个通路,其中84个差异基因注释在代谢功能类别中,而氧化磷酸化通路富集最显著。氧化磷酸化通路中,相关基因表达以上调为主(26个DEGs,20个上调,6个下调)。qRT-PCR验证结果与转录组测序结果的表达变化趋势一致。【结论】波吉卵囊藻滤液引起铜绿微囊藻氧化磷酸化途径上基因表达上调,ATP合成效率提高。  相似文献   

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The aquaculture of sea cucumber Apostichopus japonicus (Echinodermata, Holothuroidea) has grown rapidly during recent years and has become an important sector of the marine industry in Northern China. However, with the rapid growth of the industry and the use of non-standard culture techniques, epidemic diseases of A. japonicus now pose increasing problems to the industry. To screen the genes with stress response to bacterial infection in sea cucumber at a genome wide level, we constructed a cDNA library from A. japonicus Selenka (Aspidochirotida: Stichopodidae) after infecting them with Vibrio sp. for 48 h. Total RNA was extracted from the intestine, mesentery and coelomocyte of infected sea cucumber using Trizol and mRNA was isolated by Oligotex mRNA Kits. The ligated cDNAs were transformed into DH5α, and a library of 3.24×105 clones (3.24×105 cfu mL-1) was obtained with the sizes of inserted fragments ranging from 0.8 to 2.5 kb. Sequencing the cDNA clones resulted in a total of 1106 ESTs that passed the quality control. BlastX and BlastN searches have identified 168 (31.5%) ESTs sharing significant homology with known sequences in NCBI protein or nucleotide databases. Among a panel of 25 putative immunity-related genes, serum lectin isoform, complement component 3, complement component 3-like genes were further studied by real-time PCR and they all increased more than 5 fold in response to Vibrio sp. challenge. Our library provides a valuable molecular tool for future study of invertebrate immunity against bacterial infection and our gene expression data indicates the importance of the immune system in the evolution and development of sea cucumber.  相似文献   

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We developed a species-specific PCR method to identify species among dehydrated products of 10 sea cucumber species. Ten reverse species-specific primers designed from the 16S rRNA gene, in combination with one forward universal primer, generated PCR fragments of ca. 270 bp length for each species. The specificity of the PCR assay was tested with DNA of samples of 21 sea cucumber species. Amplification was observed in specific species only. The species-specific PCR method we developed was successfully applied to authenticate species of commercial products of dehydrated sea cucumber, and was proven to be a useful, rapid, and low-cost technique to identify the origin of the sea cucumber product.  相似文献   

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The exceptionally large individual growth variation has been previously recognized in several sea cucumber cohorts. However, there is a lack of information regarding the mechanism of such individual differences. In this study, the sea cucumber Apostichopus japonicus(Selenka)was reared individually in order to eliminate possible effects of social interaction, stocking density, etc. The results showed that there were substantial differences in growth among the sea cucumber individuals during the 100-day experiment. The special growth rate of the sea cucumber individuals differed by up to three folds(from 0. 40% to 1.01%), and the coefficient of variation in body weight increased from 12. 04% to 40. 51%. The final wet body weight, food intake and food conversion efficiency for each sea cucumber were generally positively correlated with their initial wet body weight(P<0. 05). Energy budget of the animals showed that the food energy spent on respiration was much greater(about four folds)but energy deposited for growth was much less for(initially)smaller than for larger A. Japonicus. The present result implies that there are obvious genetic differences among the sea cucumber individuals, largely accounting for the individual growth variation of the cohort sea cucumber. These results will provide some basic data for promoting selective breeding and farming of the sea cucumber.  相似文献   

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Marine microalga Nannochloropsis oceanica LAMB0001 were domesticated(-73 0 generations,-two days each)to adapt freshwater BGll medium.A number of freshwater medium adapted colonyderived strains were obtained.The strains were verified phylogenetically to be N.oceanica LAMB0001 based on the 18 S ribosomal RNA gene.Freshwater-medium adapted strain(FA1)grew faster in the BG11 medium prepared with freshwater than wild-type N.oceanica grew in f/2 medium prepared with seawater.We assumed that(1)the expression patterns of the genes that expressed differentially between FA1 and the wild-type N.oceanica exposing to the BG11 medium(WT-F)have been reprogrammed;(2)the physiological processes in which these genes involved have been modified;and(3)a Gene Ontology(GO)term or a KEGG pathway enriched by DEGs between FA1 and WT-F has been up-or down-regulated if it was enriched simultaneously by up-or down-regulated DEGs between FA1 and WT-F,respectively.Under these assumptions,we found that FA1 reprogrammed the expression patterns of a set of genes that involved in cell adhesion,membrane and membrane integrity,material transportation,cell movement,and cellular signaling network.These changes in cellular functions and metabolic pathways indicate that the microalga modified its gene expression pattern in a wide function range and at a high regulation rank in order to adapt to the freshwater medium.It is feasible to domesticate marine microalgae to a freshwater habitat,which may aid to modify their cultivation performances.  相似文献   

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Skin ulceration syndrome(SUS) is the main diseases affected the development of sea cucumber(Apostichopus japonicus) culture industries.To better observe the changes in the sea cucumber A.japonicus with SUS and understand the pathogenesis of the disease,activities of superoxide dismutase(SOD),catalase(CAT),and level of malondialdehyde(MDA) in coelomic fluid were detected using the Assay Kit and metabolite s in the body wall were assessed using ultra-performance liquid chromatography and quadrupoletime of flight mass spectrometry.The re sults indicated that level of MDA was increased during SUS compared with healthy individuals(P0.01),but activities of SOD and CAT were reduced(P0.05).In metabolomics analysis,metabolites,such as adenosine,choline,betaine aldehyde,palmitic acid,and taurine,were found to be upregulated and 2-oxoadipic acid,anthranilic acid(vitamin L1),thioetheramide-PC,cholesterol-3-sulfate,and pentadecanoic acid were downregulated(VIP1 and P0.1).Pathway enrichment analysis indicated most enrichment of KEGG pathways were mainly related to energy metabolism,immunity,and osmoregulation such as ABC transporters,glycine,serine and threonine metabolism,tryptophan metabolism and neuroactive ligand-receptor interaction.Our study reflected the difference in enzyme activity and metabolites between A.japonicus with SUS and those without,which will provide reference data for investigating SUS.  相似文献   

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Monthly changes in sedimentation and sediment properties were studied for three different culture treatments: sea cucumber monoculture (Mc), sea cucumber and scallop polyculture (Ps-c) and scallop monoculture (Ms). Results indicated that the survival rate of sea cucumber was significantly higher in Ps-c cultures than in Mc cultures. Sea cucumber yield was 69.6% higher in Ps-c culture than in Mc culture. No significant differences in body weight and scallop shell length were found between Ps-c and Ms cultures. The mean sedimentation rate of total particulate matter (TPM) was 72.2 g/(m2 d) in Ps-c cultures, with a maximum of 119.7 g/(m2 d), which was markedly higher than that of Mc (mean value). Sedimentation rates of organic matter (OM), total organic carbon (TOC), total nitrogen (TN) and total phosphorus (TP) in Ps-c cultures were also significantly higher than those in Mc cultures. TOC and TN contents of sediment increased rapidly in the first 5 months in Ms cultures and remained at a high level. TOC and TN contents in Mc and Ps-c cultures decreased during sea cucumber feeding seasons and increased during sea cucumber dormancy periods (summer and winter). The study demonstrates that co-culture of sea cucumber and scallop in earthen ponds is an alternative way to alleviate nutrient loads and improve water quality in coastal aquaculture systems. Moreover, it provides the additional benefit of an increased sea cucumber yield.  相似文献   

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The growth, metabolism and physiological response of the sea cucumber, Apostichopus japonicus, were investigated during periods of inactivity. The body weight, oxygen consumption rate (OCR), activities of acidic phosphatase (ACP), alkaline phosphatase (AKP), catalase (CAT) and superoxide dismutase (SOD), and content of heat shock protein 70 (Hsp70) in the body wall and coelomic fluid of A. japonicus were measured during starvation, experimental aestivation and aestivation. The results showed that the body weight of sea cucumber in the three treatments decreased significantly during the experimental period (P < 0.05). The OCR of sea cucumber reduced in starvation and experimental aestivation treatments, but increased gradually in natural aestivation treatment. The activities of ACP and AKP of sea cucumber decreased gradually in all treatments, whereas those of SOD and CAT as well as Hsp70 content decreased in the starvation and experimental aestivation treatments and increased in natural aestivation treatment. The sea cucumber entered a state of aestivation at 24℃. To some extent, the animals in experimental aestivation were different from those in natural aestivation in metabolism and physiological response. These findings suggested that the aestivation mechanism of A. japonicus is complex and may not be attributed to the elevated temperature only.  相似文献   

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This study investigated the bacterial diversity of gut content of sea cucumber(Apostichopus japonicus) and its habitat surface sediment in a bottom enhancement area using PCR-based denaturing gradient gel electrophoresis(DGGE) technique. Bacterial diversity evaluation showed that the value of the Shannon-Wiener index of gut content in different intestinal segments of A. japonicus varied between 2.88 and 3.00, lower than that of the surrounding sediment(3.23). Phylogenetic analysis showed that bacterial phylotypes in gut content and the surrounding sediment of A. japonicus were closely related to Proteobacteria includingγ-, α-, δ-and ε-proteobacteria, Bacteroidetes, Firmicute, and Actinobacteria, of which γ-proteobacteria were predominant. These results suggested that the sea cucumber A. japonicus was capable of feeding selectively, and PCR-DGGE was applicable for characterizing the bacterial community composition in gut content and the surrounding sediment of sea cucumber. Further investigation targeting longer 16S rDNA gene fragments and/or functional genes was recommended for obtaining more information of the diversity and function of bacterial community in the gut content of sea cucumber.  相似文献   

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A marine bacterium, Pseudoalteromonas sp. BC228 was supplemented to feed in a feeding experiment aiming to determine its ability of enhancing the digestive enzyme activity and immune response of juvenile Apostichopus japonicus. Sea cucumber individuals were fed with the diets containing 0(control), 105, 107 and 109 CFU g-1 diet of BC228 for 45 days. Results showed that intestinal trypsin and lipase activities were significantly enhanced by 107 and 109 CFU g-1 diet of BC228 in comparison with control(P 0.01). The phagocytic activity in the coelomocytes of sea cucumber fed the diet supplemented with 107 CFU g-1 diet of BC228 was significantly higher than that of those fed control diet(P 0.05). In addition, 105 and 107 CFU g-1 diet of BC228 significantly enhanced lysozyme and phenoloxidase activities in the coelomic fluid of sea cucumber, respectively, in comparison with other diets(P 0.01). Sea cucumbers, 10 each diet, were challenged with Vibrio splendidus NB13 after 45 days of feeding. It was found that the cumulative incidence and mortality of sea cucumber fed with BC228 containing diets were lower than those of animals fed control diet. Our findings evidenced that BC228 supplemented in diets improved the digestive enzyme activity of juvenile sea cucumber, stimulated its immune response and enhanced its resistance to the infection of V. splendidus.  相似文献   

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Crossbreeding is an effective approach to manage the genetic decline in aquaculture.One-way hybrids of Crassostrea sikamea(♀)and Crassostrea gigas(♂)have advantages in growth traits and adaptation to high temperature.Here,we used high-throughput sequencing to analyze the molecular processes in the hybrids under and after thermal stress.The hybrids were cultured in the seawater with an increasing temperature from 25℃to 40℃during 10 hours,which is regarded as the thermal stress stage.Then the temperature decreased from 40℃to 25℃within 2 h,which is regarded as the recovery stage.In this study,1293 significant diffe-rentially expressed genes(DEGs)were obtained under thermal stress,of which 576 were upregulated and 717 were downregulated,and 740 significant differentially expressed genes(DEGs)were obtained in the recovery stage,of which the number of upregulated and downregulated genes was 409 and 331,respectively.The antigen processing and presentation,NOD-like,and NF-kappa B path-ways were significantly enriched during the thermal stress stage.The MAPK and PPAR signaling pathways were significantly enrich-ed during the recovery stage.The HSP70,HSP90,and CANX genes were strongly and rapidly upregulated in the control/thermal stress groups but were slightly less upregulated in the thermal stress/recovery group.These results indicate that the innate immune system or nonspecific immunity was deployed to protect interior tissues from thermal stress.In addition,85%of the mutual DEGs were involved in bidirectional regulation(up/down or down/up)when the oysters were removed from the thermal stress to recover.This study provides preliminary insight into the molecular response of C.sikamea(♀)and C.gigas(♂)hybrids to thermal stress and provides a basis for future studies on temperature-adaptation and the possible expansion of hybrid breeding.  相似文献   

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