Institutional Repository of South China Sea Institute of Oceanology, CAS
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Elevated CO2 delays the early development of scleractinian coral Acropora gemmifera
No full textThe effects of elevated CO2 on the early life stages of coral were investigated by culturing the pelagic larvae and new recruits of Acropora gemmifera at three concentrations of CO2 (corresponding to pH = 8.1, 7.8 and 7.5, respectively). Acidified seawater resulted in fewer A. gemmifera larvae settling, and led to the production of smaller new recruits by slowing the development of the skeleton. The delayed development of new recruits due to elevated CO2 was consistent with the downregulation of calcification related genes. Several genes related to HCO3- and Ca2+ transporters were downregulated by elevated CO2, with solute carriers (SLC) (membrane transport proteins) possibly playing an important role. The downregulation of these membrane transport proteins might suppress the transport of calcium, bicarbonate and organic matter, resulting in the delayed development of A. gemmifera
A new L-type lectin (LvLTLC1) from the shrimp Litopenaeus vannamei facilitates the clearance of Vibrio harveyi
No full textAccording to structures and functions of lectins found in shrimp, they are classified into seven types, namely, L-type, C-type, P-type, M-type, galectins, fibrinogen-like domain lectins, and calnexin/calreticulin. Until now, the researches of shrimp lectins are mainly focused on C-type lectins. In this study, we identified a new L-type lectin, designated as LvLTLC1, from the shrimp Litopenaeus vannamei. The cDNA of LvLTLC1 is 1184 bp with an open reading frame of 990 bp encoding a protein of 329 amino acids. The LvLTLC1 protein contained a putative signal peptide, an L-type lectin-like domain, and a transmembrane helix region. Phylogenetic analysis showed that LvLTLC1 belonged to VIP36-like family. LvLTLC1 was expressed in all examined tissues but had higher expression level in gills and hepatopancreas than other tissues. LvLTLC1 expression was up-regulated after immune challenge by Vibrio harveyi and lipopolysaccharide. The recombinant LvLTLC1 agglutinated Gram-positive (Staphylococcus aureus) and Gram-negative bacteria (V. harveyi, V. parahaemolyticus, V. alginolyticus, V. cholerae, V. vulnificus, Pseudomonas aeruginosa, P. fluorescens) in a calcium-independent manner. Recombinant LvLTLC1 exerted the ability of enhancing the clearance of V. harveyi injected in shrimp. Our results indicated that LvLTLC1 functions in anti-pathogen innate immunity of shrimp
Community Composition and Transcriptional Activity of Ammonia-Oxidizing Prokaryotes of Seagrass Thalassia hemprichii in Coral Reef Ecosystems
No full textSeagrasses in coral reef ecosystems play important ecological roles by enhancing coral reef resilience under ocean acidification. However, seagrass primary productivity is typically constrained by limited nitrogen availability. Ammonia oxidation is an important process conducted by ammonia-oxidizing archaea (AOA) and bacteria (AOB), yet little information is available concerning the community structure and potential activity of seagrass AOA and AOB. Therefore, this study investigated the variations in the abundance, diversity and transcriptional activity of AOA and AOB at the DNA and transcript level from four sample types: the leaf, root, rhizosphere sediment and bulk sediment of seagrass Thalassia hemprichii in three coral reef ecosystems. DNA and complementary DNA (cDNA) were used to prepare clone libraries and DNA and cDNA quantitative PCR (qPCR) assays, targeting the ammonia monooxygenase-subunit (amoA) genes as biomarkers. Our results indicated that the closest relatives of the obtained archaeal and bacterial amoA gene sequences recovered from DNA and cDNA libraries mainly originated from the marine environment. Moreover, all the obtained AOB sequences belong to the Nitrosomonadales cluster. Nearly all the AOA communities exhibited higher diversity than the AOB communities at the DNA level, but the qPCR data demonstrated that the abundances of AOB communities were higher than that of AOA communities based on both DNA and RNA transcripts. Collectively, most of the samples shared greater community composition similarity with samples from the same location rather than sample type. Furthermore, the abundance of archaeal amoA gene in rhizosphere sediments showed significant relationships with the ammonium concentration of sediments and the nitrogen content of plant tissue (leaf and root) at the DNA level (P < 0.05). Conversely, no such relationships were found for the AOB communities. This work provides new insight into the nitrogen cycle, particularly nitrification of seagrass meadows in coral reef ecosystems
The Vibrio alginolyticus T3SS effectors, Val1686 and Val1680, induce cell rounding, apoptosis and lysis of fish epithelial cells
No full textVibrio alginolyticus is a Gram-negative bacterium that is an opportunistic pathogen of both marine animals and people. Its pathogenesis likely involves type III secretion system (T3SS) mediated induction of rapid apoptosis, cell rounding and osmotic lysis of infected eukaryotic cells. Herein, we report that effector proteins, Val1686 and Val1680 from V. alginolyticus, were responsible for T3SS-mediated death of fish cells. Val1686 is a Fic-domain containing protein that not only contributed to cell rounding by inhibiting Rho guanosine triphosphatases (GTPases), but was requisite for the induction of apoptosis because the deletion mutant (Delta val1686) was severely weakened in its ability to induce cell rounding and apoptosis in fish cells. In addition, Val1686 alone was sufficient to induce cell rounding and apoptosis as evidenced by the transfection of Val1686 into fish cells. Importantly, the Fic-domain essential for cell rounding activity was equally important to activation of apoptosis of fish cells, indicating that apoptosis is a downstream event of Val1686-dependent GTPase inhibition. V. alginolyticus infection likely activates JNK and ERK pathways with sequential activation of caspases (caspase-8/-10, -9 and -3) and subsequent apoptosis. Val1680 contributed to T3SS-dependent lysis of fish cells in V. alginolyticus, but did not induce autophagy as has been reported for its homologue (VopQ) in V. parahaemolyticus. Together, Val1686 and Val1680 work together to induce apoptosis, cell rounding and cell lysis of V. alginolyticus-infected fish cells. These findings provide new insights into the mechanism of cell death caused by T3SS of V. alginolyticus
Xanthones and Quinolones Derivatives Produced by the Deep-Sea-Derived Fungus Penicillium sp SCSIO Ind16F01
No full textChemical investigation of the fungus Penicillium sp. SCSIO Ind16F01 derived from deep-sea sediment sample afforded a new xanthone, 3,8-dihydroxy-2-methyl-9-oxoxanthene-4-carboxylic acid methyl ester (1) and a new chromone, coniochaetone J (2), together with three known xanthones, 8-hydroxy-6-methyl-9-oxo-9H-xanthene-1-carboxylic acid methyl ester (3), 7,8-dihydroxy-6-methyl-9-oxo-9H-xanthene-1-carboxylic acid methyl ester (4), 1,6,8-trihydroxy-3-(hydroxymethyl)anthraquinone (5), three known chromones, coniochaetone B (6), citrinolactones B (7), epiremisporine B (8), and four reported rare class of N-methyl quinolone lactams: quinolactacins B (9), C1 (10), and C2 (11), and quinolonimide (12). The structures of new compounds were determined by analysis of the NMR and MS spectroscopic data. Those isolated compounds were evaluated for their antiviral (EV71 and H3N2) and cytotoxic activities
Leptin Stimulates Prolactin mRNA Expression in the Goldfish Pituitary through a Combination of the PI3K/Akt/mTOR, MKK3/6/p(38)MAPK and MEK1/2/ERK1/2 Signalling Pathways
No full textLeptin actions at the pituitary level have been extensively investigated in mammalian species, but remain insufficiently characterized in lower vertebrates, especially in teleost fish. Prolactin (PRL) is a pituitary hormone of central importance to osmoregulation in fish. Using goldfish as a model, we examined the global and brain-pituitary distribution of a leptin receptor (lepR) and examined the relationship between expression of lepR and major pituitary hormones in different pituitary regions. The effects of recombinant goldfish leptin-AI and leptin-AII on PRL mRNA expression in the pituitary were further analysed, and the mechanisms underlying signal transduction for leptin-induced PRL expression were determined by pharmacological approaches. Our results showed that goldfish lepR is abundantly expressed in the brain-pituitary regions, with highly overlapping PRL transcripts within the pituitary. Recombinant goldfish leptin-AI and leptin-AII proteins could stimulate PRL mRNA expression in dose-and time-dependent manners in the goldfish pituitary, by both intraperitoneal injection and primary cell incubation approaches. Moreover, the PI3K/Akt/mTOR, MKK3/6/p(38)MAPK, and MEK1/2/ERK1/2-but not JAK2/STAT 1, 3 and 5 cascades-were involved in leptin-induced PRL mRNA expression in the goldfish pituitary
Fish DDX3X exerts antiviral function against grouper nervous necrosis virus infection
No full textHuman DEAD box ATP-dependent RNA helicase DDX3X has been demonstrated to exert crucial functions in carcinogenesis and antiviral immune response. However, to our knowledge, few information focused on the functions of fish DDX3X. In this study, we cloned and characterized a DDX3X homolog from orange spotted grouper (Epinephelus coioides) (EcDDX3X). EcDDX3X encoded a 733-amino acid protein which shared 97% and 76% identity to spiny damselfish (Acanthochromis polyacanthus) and human (Homo sapiens), respectively. Amino acid alignment analysis showed that EcDDX3X contained conserved DExDc and Helic C domains. The transcription levels of EcDDX3X were significantly increased in poly I:C transfected cells and red-spotted grouper nervous necrosis virus (RGNNV) infected cells. Under fluorescence microscopy, the green fluorescence was observed evenly in the cytoplasm in EcDDX3X transfected cells. The ectopic expression of EcDDX3X significantly inhibited the replication of RGNNV, evidenced by the decreased numbers of the vacuoles evoked by RGNNV infection, and the reduced transcription levels of RGNNV coat protein (CP) and RNA-dependent RNA polymerase (RdRp) genes. In contrast, the replication of Singapore grouper iridovirus (SGIV) in grouper spleen (GS) cells was not significantly affected by EcDDX3X overexpression. Further studies showed that overexpression of EcDDX3X in vitro significantly increased the expression levels of several interferon associated cytokines or effectors. Moreover, the regulatory effect of EcDDX3X on interferon immune response was dependent on its N terminal region, but not the DExDc and Helic C domain. In addition, we also found that overexpression of EcDDX3X significantly increased the interferon promoter activity, and the activation of interferon immune response was regulated by both IRF3 and IRF7. Together, our results firstly showed that fish DDX3X exerted crucial roles in antiviral immunity against RNA virus infection via upregulating interferon antiviral responses. (C) 2017 Elsevier Ltd. All rights reserved
Expression and functional characterization of TRIF in orange-spotted grouper (Epinephelus coioides)
No full textAntiviral immune responses are triggered by the innate immune recognition of viral infection. Toll/interleukin-1 receptor (TIR) domain containing adapter inducing interferon-beta (TRIF) is an adapter in responding to activation of Toll-like receptors, which provides early clearance of viral pathogens. Our study focuses on the functional characterization of grouper TRIF (EcTRIF) based on the comparison of its sequence and functional evolution from grouper fish to mammals. The results show that the open reading frame of EcTRIF encoded a protein of 580 amino acids. Real-time PCR analysis indicates that EcTRIF was constitutively expressed in all the analyzed tissues in healthy grouper. EcTRIF was significantly induced in spleen post-LPS and poly (I:C) stimulation. Fluorescence microscopy shows that EcTRIF is colocalized with a Golgi apparatus marker, implying its unique subcellular localization in the Golgi apparatus. Luciferase reporter assays confirmed that EcTRIF was able to activate the IFN and NF-kappa B promoter. Overexpression of EcTRIF in grouper brain cells inhibited the replication of red-spotted grouper nervous necrosis virus (RGNNV). These results indicate that EcTRIF plays an important role in modulating antiviral innate immune responses. Our results have applications in functional studies on TRIF in teleost fish and immune evolution. (C) 2017 Elsevier Ltd. All rights reserved
Vibrio alginolyticus 16S-23S intergenic spacer region analysis, and PCR assay for identification of coral pathogenic strain XSBZ03
No full textPorites andrewsi white syndrome (PAWS), caused by Vibrio alginolyticus strains XSBZ03 and XSBZ14, poses a serious threat to corals in the South China Sea. To obtain a specific target against which to develop a rapid PCR detection method for the coral pathogenic strain XSBZ03, the 16S-23S rRNA gene intergenic spacer (IGS) region of 4 strains of V. alginolyticus, including the XSBZ03 and XSBZ14 strains, was amplified, sequenced and analyzed. Six types of IGS were found: IGS(0), IGS(G), IGS(IA), IGS(AG), IGS(GLV), and IGS(GLAV). IGS(0), IGS(G), IGS(IA), IGS(AG) and IGS(GLV) appeared to be the most prevalent forms in the 4 strains and the percentage identity range within each type was 91.4-100%, 89.3-98.5%, 83.0-99.8%, 91.5-95.6%, and 88.7-99.3%, respectively. IGS(GLAV) was found only in the HN08155 strain, a causative agent of fish disease. IGS(GLAV), IGS(GLV) and IGS(AG) are reported here for the first time in V. alginolyticus. An IGS sequence specific to the XSBZ03 strain was identified following alignment of the homologous IGSs, and used to design strain-specific primers for its rapid identification by PCR. The results from PCR analysis suggest that the method is a rapid, practical, and reliable tool for the identification of the XSBZ03 strain in samples of isolated bacteria, as well as seawater and coral samples spiked with the bacterial strain. This is the first report of a rapid diagnostic assay for a causative agent of PAWS, based on PCR detection of a coral pathogen at the strain level. After applying this assay in coral transplantation, the survival rates of transplanted corals were significantly increased. This diagnostic assay should aid with both the elucidation of the cause of the disease, and transplantation of PAWS-free P. andrewsi in the South China Sea
Nitrogenous Compounds Produced by the Deep Sea Derived Fungus Leptosphaeria sp SCSIO 41005
No full textFour nitrogenous compounds, including two new compounds methyl-4-((1-hydroxy-3-methylpentan-2-yl)amino)-3-methyl-4-oxobutanoate (1), 4-((1-acetoxy-3-methylpentan-2-yl)amino)-3-methyl-4-oxobutanoic acid (2), and two amino acid derivatives (3, 4), were isolated from the deep sea derived fungus Leptosphaeria sp. SCSIO 41005, together with a cyclohexanone derivative (5) as new natural compound and other three known compounds (6 8). Their structures were elucidated by means of comprehensive 1D, 2D NMR and HR-ESI-MS spectroscopic methods. All the compounds were evaluated for their cytotoxic and antiviral activities