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中国沿海污损性海鞘生态特点及研究展望
海鞘是海洋污损生物群落的重要组成部分,其附着会给水产养殖业带来严重危害。在中国沿海,引发生物污损的海鞘共8科20属40种,其中渤海海域7种,黄海17种,东海23种和南海27种。北方海域以柄瘤海鞘(Styela clava)、乳突皮海鞘(Molgula manhattensis)和米氏小叶鞘(Diplosoma listerianum)为优势种,南方则以冠瘤海鞘(Styela canopus)、皱瘤海鞘(S.plicata)和大洋纵列海鞘(Symplegma oceania)为优势种。海鞘污损特点与设施所处地理位置、浸海时间、环境状况等因子密切有关。今后除了继续对沿岸代表性海域开展深入研究外,还需结合海洋经济发展和科学研究的需要,加强离岸设施(尤其深海环境)污损生物群落的研究,以期更好地掌握污损性海鞘的组成分布、种群特点及发展趋势
5科11种鱼类ITS1特征分析及其在系统分类研究中的适用性
为了探讨ITS1作为分子标记用于鱼类系统演化的适用性,实验选取鲈形目5科11种鱼类为研究对象,包括尖吻鲈科、射水鱼科、军曹鱼科、剑鱼科和鲹科。通过克隆和测序等技术共获得了348条ITS1序列,长度范围为442~661 bp;通过对所有序列的长度、变异位点数量、GC含量、核苷酸多样性及单倍型多样性指数等遗传特征比较分析发现,11种鱼类ITS1序列无论是在种内还是在种间,长度和序列都表现出较为明显的多态性。特别是在军曹鱼中,70条克隆的长度范围为648~661 bp,但有一条序列存在55 bp缺失,结合该序列的GC含量,二级结构和最小自由能,推断该序列为假基因。以鮣为外类群,基于核糖体ITS1序列构建的邻接树显示在物种种类水平上,不同个体的克隆都按种类聚支,ITS1可以用于该类群物种的区分;在属级水平上,ITS1将11属鱼类完全区分开,能够用于属级水平的区分;在科级水平上,虽然鲹科分为2支的分子结果和形态分类存在差异,但ITS1构建的系统关系与线粒体分子标记构建的系统进化树相似。研究表明,核糖体ITS1可以作为一种有效的分子标记用于研究鱼类的系统分类研究,并且不同的分类阶元其解析能力不同,这将为鱼类核糖体的研究提供科学依据
TBK1 from orange-spotted grouper exerts antiviral activity against fish viruses and regulates interferon response
TANK-binding kinase-1 (TBK1) has been well studied in mammals because of its importance in type I interferon induction in antiviral immunity. However, the roles of fish TBK1 in virus infection still remained largely uncertain. In the current study, a TBK1 homolog from orange-spotted grouper (Epinephelus coioides) (EcTBK1) was cloned and its roles in fish viral infections were investigated. Sequence analysis showed that EcTBK1 encoded a 723-amino acid peptide which shared 98% and 73% identity to large yellow croaker (Larimichthys crocea) and human (homo sapiens), respectively. Multiple sequence alignments indicated that EcTBK1 contained conserved domains, including N-terminal kinase domain (KD), the middle ubiquitin-like domain (ULD) and C-terminal coiled-coil (CC) domains. The tissue distribution profiles demonstrated that EcTBK1 gene was constitutively expressed in all examined tissues, with predominant expression in intestine. Temporal expression analysis in vitro showed that the expression levels of EcTBK1 were significantly up-regulated in response to both red-spotted grouper nervous necrosis virus (RGNNV) and Singapore grouper iridovirus (SGIV) infection, suggested that EcTBK1 might exert crucial roles in fish virus infection. Subcellular localization indicated that EcTBK1 expression was primarily in the cytoplasm in GS cells. The ectopic expression of EcTBK1 significantly inhibited both SGIV and RGNNV replication. Furthermore, EcTBK1 overexpression significantly increased the expression levels of interferon related cytokines and pro-inflammatory factors. In addition, the overexpression of EcTBK1 increased the IRF3- and IRF7-regulated interferon promoter ISRE and IFN activity, and the regulatory effect on interferon immune response were dependent on its kinase domain. Together, we speculated that grouper TBK1 exerted antiviral activity against iridovirus and nodavirus via regulating the interferon immune and inflammatory response
Establishment and characterization of a kidney cell line from kelp grouper Epinephelus moara
A novel cell line, Epinephelus moara kidney cell line (EMK), was established from kidneys of kelp grouper E. moara. Cells were cultured at 24 A degrees C in Leibovitz's L-15 medium (L15) supplemented with antibiotics, basic fibroblast growth factor (bFGF), foetal bovine serum (FBS) and 2-mercaptoethanol (2-ME). EMK cells, fibroblastic in morphology, proliferated to 100% confluency in 3-4 days and were subcultured for over 50 passages. The cells could grow from 18 to 30 A degrees C, with optimal growth at 24 A degrees C. Chromosome analysis indicated that the modal chromosome number was 48 in the cells at passage 42. Green fluorescent signals could be observed in EMK cells when the cells were transfected with pEGFP-N3 plasmid. Moreover, a significant cytopathic effect (CPE) was observed in the cells after infection with Singapore grouper iridovirus (SGIV) or nervous necrosis virus (NNV), and viral replication was confirmed by quantitative real-time PCR (qPCR). These results suggested the potential of the EMK cell line for studies of transgene and pathogenesis of SGIV and NNV
Isochromophilones A-F, Cytotoxic Chloroazaphilones from the Marine Mangrove Endophytic Fungus Diaporthe sp SCSIO 41011
Six new highly oxygenated chloroazaphilone derivatives, isochromophilones A-F (1-6), were obtained from the mangrove-derived fungus Diaporthe sp. SCSIO 41011, together with six known analogues (7-12). The structures of 1-6 including absolute configurations were determined by detailed NMR, MS spectroscopic analyses, and electronic circular dichroism spectra. Compounds 1 and 2 represent the first reported azaphilones lacking a carbonyl group at C-6. Compound 8 exhibited cytotoxic activities against three renal carcinoma cell lines, ACHN, OS-RC-2, and 786-O cells, with IC50 values ranging from 3.0 to 4.4 mu M, and 4 showed activity against 786-O cells with an ICso of 8.9 mu M. Further studies indicated that 4 induced apoptosis in 786-O cells in a dose- and time-dependent manner
Strand-specific RNA-Seq analysis provides first insight into transcriptome response of & IT;Vibrio alginolyticus & IT; to phage infection
RNA-Seq transcriptome analysis of bacteria during phage infection has been occasionally reported in last several years but little is known about how Vibrio bacteria respond to phage invasion on global transcriptome level. In the present study, transcriptome profiling of V. alginolyticus during a lytic phage infection was first investigated through strand-specific RNA-Seq analysis. The results revealed that 346 unigenes were up-regulated and 860 unigenes were down-regulated during the infection of phage Vp670. Notably, the unigenes involving in phosphorelay signal transduction system were largely down-regulated in infected V. alginolyticus cells. However, some unigenes in the GO terms (DNA binding and sequence-specific DNA binding transcription factor activity) were up-regulated including a csp gene coding cold shock protein. These transcriptome data contribute to a more comprehensive insight into cellular response of V. alginolyticus to phage infection and help us deeply understand the phage-host interplay on molecular levels. (C) 2017 Elsevier B.V. All rights reserved
Homogentisic acid gamma-lactone suppresses the virulence factors of Pseudomonas aeruginosa by quenching its quorum sensing signal molecules
Bacterial quorum sensing (QS) molecules are involved in the coordination of certain behaviors such as biofilm formation, virulence and antibiotic resistance. QS molecules (autoinducers) and their corresponding receptors have been recognized as important therapeutic targets for drug -resistant infections and biofilm-associated infections (BAI). This study assessed the multiple biological effects of homogentisic acid gamma-lactone (HgAL), a furanone derivative. The anti -QS and anti-biofilm effects of HgAL against PAO1 strain were evaluated using CLSM, SEM, HPLC and other biochemical methods The results showed that HgAL could effectively inhibit the production of pyocyanin and extracellular matrix, as well as reduce the adherence ability and biofilm formation of Pseudomonas aeruginosa. Inhibition of virulence is attributed to the suppressive effect of HgAL on biosynthesis of 3-oxo-C-12-HSL and C-4-HSL (two kinds of QS signaling molecules in P. aeruginosa). Our results support HgAL as a potential agent for prevention of BAI in the healthcare settings. (C) 2017 Chinese Chemical Society and Institute of Materia Medica, Chinese Academy of Medical Sciences. Published by Elsevier B.V. All rights reserved
Growth performance and biochemical composition of the oysters Crassostrea sikamea, Crassostrea angulata and their hybrids in southern China
The Kumamoto oyster (Crassostrea sikamea) and the Portuguese oyster (C.angulata) are important aquaculture species which naturally coexist along the southern coast of China. To understand the potential feasibility of hybridization between the two species, we conducted two-by-two factorial cross-experiments in Beihai (Guangxi province), and also compared the survival and growth of the hybrids to that of the two parental progenies during the grow-out period from July 2014 to July 2015. Genetic analysis confirmed that the hybrid spats were true hybrids. Additionally, the biochemical composition of the 1-year-old oyster progenies was determined. In July 2015, the mean shell height of the hybrids was 42.98 +/- 6.29mm, which was higher than that of the Kumamoto oyster progeny. The cumulative survival rate of the hybrids was 26.37 +/- 1.32%, which was higher than that of the progeny of the Portuguese oyster. Mean lipid content of the hybrids was 13.65 +/- 1.63% of dry weight, which showed obvious heterosis compared to those of the two parental progenies. Observation of gonads revealed that all hybrids were completely fertile. Furthermore, relative expression of the lipid homeostasis genes, SREBP (sterol regulatory element-binding proteins), PPAR (peroxisome proliferator-activated receptor) and INSIG (insulin-induced gene) were found to vary between parental progenies and the hybrids, thus providing a possible reason for difference in the lipid contents of these experimental groups. Overall, the hybrids were viable, rich in lipid and completely fertile and thus could serve as a promising aquaculture stock for oyster breeding in southern China
Nitrobenzoyl Sesquiterpenoids with Cytotoxic Activities from a Marine-Derived Aspergillus ochraceus Fungus
Nitrobenzoyl sesquiterpenoids are rare from natural sources. Two new nitrobenzoyl sesquiterpenoids, insulicolide B (1) and insulicolide C (3), and the new natural product 14-O-acetylinsulicolide A (2) were isolated from culture extracts of the marine-derived fungus Aspergillus ochraceus JcmalF17, together with three known nitrobenzoyl sesquiterpenoids (4-6) and a derivative sesquiterpenoid (7). The structures of the new compounds, including their absolute configurations, were determined by NMR. and MS spectroscopic data analyses and comparison between the calculated and experimental ECD spectra. The nitrobenzoyl sesquiterpenoids (1-6) were evaluated for their cytotoxicities against three renal carcinoma cell lines, ACHN, OS-RC-2, and 786-O cells, and compounds 2, 4, and 5 displayed activities with IC50 values of 0.89 to 8.2 mu M. Further studies indicated that 2 arrested the cell cycle at the G0/G1 phase at a concentration of 1 mu M and induced late apoptosis at a concentration of 2 mu M after a 72 h treatment of 786-O cells
Insights into the simultaneous removal of Cr6+ and Pb2+ by a novel sewage sludge-derived biochar immobilized nanoscale zero valent iron: Coexistence effect and mechanism
Cr6+ and Pb2+ are both highly toxic pollutants and commonly co-exist in some industrial effluents and contaminated waters. In this study, simultaneous removal of Cr6+ and Pb2+ by a novel sewage sludge-derived biochar immobilized nanoscale zero-valent iron (SSB-nZVI) was systematically investigated. It was well demonstrated that a porous structure was successfully formed on the SSB nZVI when the starch was used as an additive. A synergistic effect on the adsorption and reduction over the SSB-nZVI was achieved, resulting in nearly 90 and 82% of Cr6+ and Pb2+ removal within 30 min, respectively. Cr6+ was reduced prior to Pb2+. A low pH could accelerate the corrosion of nZVI as well as phosphate leaching. When Malachite green was added as a coexisting organic pollutant, its effective removal was found due to the formation of a Fenton-like system. The SSB-nZVI could be run consecutively three times with a relatively satisfactory performance. Most of Cr6+ was converted into Cr2O3 and Cr(OH)(3) on the SSB nZVI surface, whereas most of Pb2+ species existed as Pb(OH)(2) (or PbO). A possible reaction mechanism on the SSB-nZVI involved the adsorption, reduction and precipitation of both Cr6+ and Pb2+ over the particles. Present study sheds light on the insight of the fate and transport of Cr6+ and Pb2+ in aquatic environment, as well provides helpful guide for the remediation of coexistence of pollutants in real applications. (C) 2018 Elsevier B.V. All rights reserved