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In the present study, the expression of some immune-related genes was examined as indicator to understand the development of immune defense system during the ontogenesis of scallop Chlamys farreri. The mRNA transcripts of pattern-recognition receptors (PRRs) including CfPGRP-S1, CfLGBP, CfLec-1 and CfLec-3 were observed at a low level or even undetected at early developmental stages from eggs to blastula, and then began to increase overwhelmingly in trochophore. For the genes of immune effector including CfLYZ, CfLBP/BPI, CfSOD and CfCAT, their mRNA transcripts were higher expressed in embryos, and increased significantly in D-hinged or early veliger larvae. The whole-mount immunofluorescence assay revealed two immunoreactive spots of CfPGRP-S1 were first observed in the mid-ventral region of prototroch in trochophore, and the immunopositive fluorescence of CfLGBP, CfLec-1 and CfLec-3 appeared at the same spots in early D-hinged larvae. Most of the PRRs were located in velum, mouth, esophagus and stomach region in early and mid-veliger larvae, and especially the strong immunopositive fluorescence of CfLec-3 was observed in velum. The immunoreactive areas of CfLYZ, CfLBP/BPI, CfSOD and CfCAT were observed in trochophore and early D-hinged larvae. After D-hinged larvae, they distributed in different tissues from the edge of velum, mouth, esophagus to the region around digestive gland. After bacterial challenge, the mRNA expression of CfLGBP, CfLec-1 and CfLec-3 did not change significantly in trochophore, while a down-regulation of CfPGRP-S1 was observed at 6 h (P < 0.05). The expression of CfPGRP-S1 and CfLGBP decreased or increased inversely in D-hinged and late veliger larvae respectively, whereas CfLec-1 and CfLec-3 increased significantly during 6-24 h after bacterial challenge in the two stages (P < 0.05). In contrast, the expressions of immune effectors in trochophore and late veliger larvae were significant up-regulated at 6 h, 12 h or 24 h after bacterial challenge (P < 0.05). However, in late D-hinged larvae, CfLYZ and CfSOD expressions were significantly down-regulated at 6 h, while CfLBP/BPI expression was up-regulated at 6 h and 24 h post challenge (P < 0.05). These results indicated that the immune defense system of scallop might appear firstly in the mid-ventral region of prototroch in trochophore, and developed maturely after late D-hinged larvae. The developing immune system in the D-hinged and late veliger larvae could respond to the immune stimulation in different manner. Copyright © 2013 Elsevier Ltd. All rights reserved.

Hatchery-reared larvae of the Pacific oyster (Crassostrea gigas) often suffer from massive mortality induced by Ostreid herpesvirus 1 (OsHV-1) infection, indicating the importance of better understanding of oyster immune defense systems. The accuracy of measurements of gene expression levels based on quantitative real-time PCR assays relies on the use of housekeeping genes as internal controls; however, few studies have focused on the selection of such internal controls. In this study, we conducted a comprehensive investigation of internal control genes during oyster development in virus-infected and uninfected samples. Transcriptome data for 38 developmental stages were downloaded and the gene expression patterns were classified into 30 clusters. A total of 317 orthologs of classical housekeeping genes in the oyster genome were annotated. After combining the expression profiles and oyster housekeeping gene dataset, 14 candidate internal controls were selected for further investigation: Elongation factor-1α (EF-1α), 18S rRNA (18S), 28S rRNA (28S), Glyceraldehyde 3-phosphate dehydrogenase (GAPDH), β-actin (ACT), Ribosomal protein L7 (RL7), Ribosomal protein L27 (RL27), Ribosomal protein L36 (RL36), Ribosomal protein S18 (RS18), Heterogeneous nuclear ribonucleoprotein A2/B1 (RO21), Eukaryotic translation elongation factor 2 (EF2), Ubiquitin-conjugating enzyme E2D2 (UBCD1), S-phase kinase-associated protein 1 (SKP1) and Heterogeneous nuclear ribonucleoprotein Q (HNRPQ). RNA was extracted from oyster larvae infected with OsHV-1 (group A; GA), and OsHV-1 free larvae (group B; GB). The expression levels of the 14 candidate internal controls were studied in GA and GB larvae by real-time PCR. Their expression stabilities were further analyzed using the GeNorm program. RL7 and RS18 were the most stable genes in both OsHV-1 infected (GA) and uninfected (GB) larvae. These results suggest that RL7 and RS18 could be used as internal controls for studying gene expression in normal growing oyster larvae and in OsHV-1 infected larvae. These high quality internal controls will be a valuable resource in future studies of oyster larval mortality. Copyright © 2012 Elsevier Ltd. All rights reserved.

Wang B.-G.,CAS Qingdao Institute of Oceanology | Gloer J.B.,University of Iowa | Ji N.-Y.,CAS Yantai Institute of Coastal Zone Research | Zhao J.-C.,CAS Qingdao Institute of Oceanology
Chemical Reviews | Year: 2013

The article highlights the diversity of halogenated organic molecules produced by marine red algal species in the family Rhodomelaceae. Although no review on the many halogenated molecules derived from marine red algae of the family Rhodomelaceae appeared in the literature up to now, a number of excellent reviews on various aspects of naturally occurring halogenated molecules were published. An excellent review dealing with the structures, biogenetic considerations, and biological activities of both halogenated and nonhalogenated polyethers from red algae and sponges appeared in 2000. Because they possess a variety of novel structures and often display potent biological activities, halogenated organic molecules have attracted considerable attention as challenging targets for partial and total synthesis. A number of elegant synthetic strategies and methodologies were developed and employed for the synthesis of such compounds.

Liu J.Y.,CAS Qingdao Institute of Oceanology
PLoS ONE | Year: 2013

China's seas cover nearly 5 million square kilometers extending from the tropical to the temperate climate zones and bordering on 32,000 km of coastline, including islands. Comprehensive systematic study of the marine biodiversity within this region began in the early 1950s with the establishment of the Qingdao Marine Biological Laboratory of the Chinese Academy of Sciences. Since that time scientists have carried out intensive multidisciplinary research on marine life in the China seas and have recorded 22,629 species belonging to 46 phyla. The marine flora and fauna of the China seas are characterized by high biodiversity, including tropical and subtropical elements of the Indo-West Pacific warm-water fauna in the South and East China seas, and temperate elements of North Pacific temperate fauna mainly in the Yellow Sea. The southern South China Sea fauna is characterized by typical tropical elements paralleled with the Philippine-New Guinea-Indonesia Coral triangle typical tropical faunal center. This paper summarizes advances in studies of marine biodiversity in China's seas and discusses current research mainly on characteristics and changes in marine biodiversity, including the monitoring, assessment, and conservation of endangered species and particularly the strengthening of effective management. Studies of (1) a tidal flat in a semi-enclosed embayment, (2) the impact of global climate change on a cold-water ecosystem, (3) coral reefs of Hainan Island and Xisha-Nansha atolls, (4) mangrove forests of the South China Sea, (5) a threatened seagrass field, and (6) an example of stock enhancement practices of the Chinese shrimp fishery are briefly introduced. Besides the overexploitation of living resources (more than 12.4 million tons yielded in 2007), the major threat to the biodiversity of the China seas is environmental deterioration (pollution, coastal construction), particularly in the brackish waters of estuarine environments, which are characterized by high productivity and represent spawning and nursery areas for several economically important species. In the long term, climate change is also a major threat. Finally, challenges in marine biodiversity studies are briefly discussed along with suggestions to strengthen the field. Since 2004, China has participated in the Census of Marine Life, through which advances in the study of zooplankton and zoobenthos biodiversity were finally summarized. © 2013 J. Y. Liu.

Wang L.,CAS Qingdao Institute of Oceanology
Developmental and comparative immunology | Year: 2013

The economical and phylogenic importance of mollusc has led an increasing number of investigations giving emphasis to immune defense mechanism. This review discusses the advances in immunological study of mollusc in China, with special reference to dominant aquaculture species over the past decades. As an invertebrate group, molluscs lack adaptive immunity and consequently they have evolved sophisticated strategies of innate immunity for defense against pathogens. This review aims to present the various immunologically significant pattern recognition receptors (PRRs), such as Toll-like receptors (TLRs), lectins, lipopolysaccharide and β-1, 3-glucan binding protein (LGBP), scavenger receptors (SRs) employed by mollucans. This work also highlights immune proteolytic cascade, TLR signaling pathway and an extensive repertoire of immune effectors including antimicrobial peptide, lysozyme, antioxidant enzyme and heat shock protein. Further, the review presents the preliminary progress made on the catecholaminergic neuroendocrine system in scallop and its immunomodulation function to throw light into neuroendocrine-immune regulatory network in lower invertebrates. Copyright © 2012 Elsevier Ltd. All rights reserved.

Li F.,CAS Qingdao Institute of Oceanology
Developmental and comparative immunology | Year: 2013

The annual production of shrimp culture in mainland of China has been over one million tons for several years. The major cultivated penaeidae species are Litopenaeus vannamei, Fenneropenaeus chinensis, Penaeus monodon and Marsupenaeus japonicus. Due to the importance of shrimp aquaculture in China, researchers have paid more attention to the molecular mechanism of shrimp disease occurrence and tried to develop an efficient control strategy for disease. This paper summarizes the research progress related to innate immunity of penaeid shrimp made in the last decade in Mainland China. Several pattern recognition receptors, such as lectin, toll, lipopolysaccharide and β-1,3-glucan binding protein (LGBP) and tetraspanin were identified. The major signal transduction pathways, including Toll pathway, IMD pathway, which might be involved in the immune response of shrimp, were focused on and most of the components in Toll pathway were identified. Also, cellular immune responses such as phagocytosis and apoptosis were regarded playing very important roles in anti-WSSV infection to shrimp. The molecules involved in the maintenance of the immune homeostasis of shrimp and the progress on molecular structure and pathogenic mechanism of WSSV were summarized. Therefore, the brief outline about the immune system of shrimp is drawn based on the recent data which will help us to understand the immune responses of shrimp to different pathogens. Copyright © 2012 Elsevier Ltd. All rights reserved.

Wang L.,CAS Qingdao Institute of Oceanology
Fish & shellfish immunology | Year: 2013

C-type lectins are a large family of Ca2+-dependent carbohydrate binding proteins which play crucial roles to recognize and eliminate pathogens in innate immunity. In the present study, a novel C-type lectin was identified from Eriocheir sinensis (designated as EsCTL). The full-length cDNA of EsCTL was of 789 bp with an open reading frame of 468 bp encoding a polypeptide of 156 amino acids with a signal sequence and single carbohydrate-recognition domain (CRD). The potential tertiary structure of the CRD adopted a typical double-loop structure with Ca2+-binding site 2 in the long loop region and two conserved disulfide bridges at the bases of the loops. An EPQ motif to determine carbohydrate binding specificity was identified in the CRD of EsCTL. The mRNA transcripts of EsCTL were mainly detected in hepatopancreas and its relative expression level in hemocytes was significantly up-regulated after the challenges of Vibrio anguillarum (P < 0.05) and Pichia pastoris (P < 0.05). The recombinant EsCTL protein (rEsCTL) could bind different PAMPs, including LPS, PGN, β-glucan, and polyI:C; and also bind various microorganisms including three Gram-positive bacteria, three Gram-negative bacteria and two yeasts. Moreover, rEsCTL could significantly enhance the in vitro encapsulation of crab hemocytes. All these results suggested that EsCTL functioned as an important PRR involved in immune defense against invading pathogen in crab. Copyright © 2012 Elsevier Ltd. All rights reserved.

The scavenger receptor cysteine-rich (SRCR) proteins are secreted or membrane-bound receptors with one or multiple SRCR domains. Members of the SRCR superfamily are known to have diverse functions that include pathogen recognition and immunoregulation. In teleost, although protein sequences with SRCR structure have been identified in some species, very little functional investigation has been carried out. In this study, we identified and characterized a teleost SRCR protein from red drum Sciaenops ocellatus. The protein was named S. ocellatus SRCR1 (SoSRCRP1). SoSRCRP1 is 410-residue in length and was predicted to be a transmembrane protein, with the extracellular region containing a collagen triple helix repeat and a SRCR domain. The SRCR domain has six conserved cysteines, of which, C338 and C399, C351 and C409, and C379 and C389 were predicted to form three disulfide bonds. SoSRCRP1 expression was detected mainly in immune-relevant tissues and upregulated by bacterial and viral infection. In head kidney leukocytes, bacterial infection stimulated the expression of SoSRCRP1, and the expressed SoSRCRP1 was localized on cell surface. Recombinant SoSRCRP1 (rSoSRCRP1) corresponding to the SRCR domain was purified from Escherichia coli and found to be able to bind Gram-negative and Gram-positive bacteria. To examine the structure-function relationship of SoSRCRP1, the mutant proteins SoSRCRP1M1, SoSRCRP1M2, SoSRCRP1M3, and SoSRCRP1M4 were created, which bear C351S and C409S, C338S, C379S, and R325A mutations respectively. Compared to rSoSRCRP1, all mutants were significantly reduced in the ability of bacterial interaction, with the highest reduction observed with SoSRCRP1M4. Taken together, these results indicate that SoSRCRP1 is a cell surface-localized SRCR protein that binds bacterial ligands in a manner that depends on the conserved structural features of the SRCR domain. Copyright © 2012 Elsevier Ltd. All rights reserved.

BACKGROUND: MicroRNAs (miRNAs) regulate gene expression by binding to mRNA transcripts in various biological processes. In mammals and birds, miRNAs are known to play vital parts in both host immune defense and viral infection. However, in lower vertebrates such as teleost, systematic investigations on host and viral miRNAs are lacking.RESULTS: In this study, we applied high-throughput sequencing technology to identify and analyze both host and viral miRNAs in Japanese flounder (Paralichthys olivaceus), an economically important teleost fish farmed widely in the world, infected with megalocytivirus at a timescale of 14 days divided into five different time points. The results showed that a total of 381 host miRNAs and 9 viral miRNAs were identified, the latter being all novel miRNAs that have no homologues in the currently available databases. Of the host miRNAs, 251 have been reported previously in flounder and other species, and 130 were discovered for the first time. The expression levels of 121 host miRNAs were significantly altered at 2 d to 14 d post-viral infection (pi), and these miRNAs were therefore classified as differentially expressed host miRNAs. The expression levels of all 9 viral miRNAs increased from 0 d pi to 10 d pi and then dropped from 10 d pi to 14 d pi. For the 121 differentially expressed host miRNAs and the 9 viral miRNAs, 243 and 48 putative target genes, respectively, were predicted in flounder. GO and KEGG enrichment analysis revealed that the putative target genes of both host and viral miRNAs were grouped mainly into the categories of immune response, signal transduction, and apoptotic process.CONCLUSIONS: The results of our study provide the first evidences that indicate existence in teleost fish (i) infection-responsive host and viral miRNAs that exhibit dynamic changes in expression profiles during the course of viral infection, and (ii) potential involvement of miRNAs in host-viral interaction.

Wang F.,CAS Qingdao Institute of Oceanology
Geophysical Research Letters | Year: 2010

A global wavenumber-3 dipole SST mode is showed to exist in the Southern Hemisphere subtropical climate variability in austral summer. A positive (negative) phase of the mode is characterized by cool (warm) SST anomalies in the east and warm (cool) SST anomalies in the southwest of the south Indian, Pacific, and Atlantic Oceans, respectively. This coherent dipole structure is largely a response of ocean mixed layer to the atmospheric forcing characterized by migration and modulation of the subtropical high-pressures, in which the latent heat flux play a leading role through wind-induced evaporation, although ocean dynamics may also be crucial in forming SST anomalies attached to the continents. Exploratory analyses suggest that this mode is strongly damped by the negative heat flux feedback, with a persistence time about three months and no spectral peak at interannual to decadal time scales. As the subtropical dipole mode is linearly independent of ENSO and SAM, whether it represents an additional source of climate predictability should be further studied. Copyright © 2010 by the American Geophysical Union.

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