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Wang Z.,Guangdong Ocean University | Wang Z.,Guangdong Provincial Key Laboratory of Aquaculture in South China Sea for Aquatic Economic Animals | Wang B.,Guangdong Ocean University | Wang B.,Guangdong Provincial Key Laboratory of Pathogenic Biology and Epidemiology for Aquatic Economic Animals | And 9 more authors.
Gene | Year: 2015

The pearl oyster Pinctada fucata is cultured widely for production of marine pearls in China, while mass mortalities, likely related to pathogenic infections, have occurred frequently in juvenile, mother and operated oysters. To address this issue, understanding host defense mechanisms of P. fucata against pathogenic challenge is extremely important. In the present study, a comparative analysis of hemocyte transcriptomes of P. fucata before and after Vibrio alginolyticus infection was conducted using the Illumina/Hiseq-2000 RNA-Seq technology. A total of 56,345,139 clean reads were generated and then assembled into 74,007 unigenes with an average length of 680. bp and an N50 of 1197. bp. Unigenes were annotated by comparing against non-redundant protein sequence (nr), non-redundant nucleotide (nt), Swiss-Prot, Pfam, Gene Ontology database (GO), Clusters of Orthologous Groups (COG) and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases, and 29,615 unigenes (40.01%) were annotated in at least one database. There were 636 genes (518 up-regulated and 118 down-regulated) that were significantly differentially expressed after bacterial challenge, and among which 369 were associated with 122 pathways, including classical immune-related pathways, such as 'MAPK signaling pathway', 'Chemokine signaling pathway', 'Apoptosis' and 'Wnt signaling pathway'. These findings provide information on the pearl oyster innate immunity and may contribute to developing strategies for management of diseases and long-term sustainability of P. fucata culture. © 2015 Elsevier B.V.


Wang Z.,Guangdong Ocean University | Wang Z.,Guangdong Provincial Key Laboratory of Aquaculture in South China Sea for Aquatic Economic Animals | Wang Z.,Guangdong Provincial Key Laboratory of Pathogenic Biology Epidemiology for Aquatic Economic Animals | Jian J.,Guangdong Ocean University | And 7 more authors.
Marine Genomics | Year: 2011

The cDNA of a tandem-repeat galectin from the pearl oyster Pinctada fucata (designated PfGal) was cloned by expressed sequence tag (EST) and rapid amplification of cDNA ends (RACE) techniques. The full-length cDNA of PfGal was 1386. bp, consisting of a 5' untranslated region (UTR) of 26. bp, a 3' UTR of 313. bp, and an open reading frame (ORF) of 1047. bp encoding a polypeptide of 348 amino acids with a predicted molecular weight of 38.09. kDa and theoretical isoelectric point of 8.49. Similar to other tandem-repeat galectins, PfGal contained two tandem carbohydrate recognition domains (CRDs), with typical conserved motifs which were important for carbohydrate recognition, and it appeared to possess neither a signal peptide nor a transmembrane domain. Fluorescent quantitative real-time PCR analyses indicated that PfGal mRNA was highly expressed in hemocytes, digestive gland and mantle, and its expression was increased in all studied tissues after Vibrio alginolyticus challenge. The temporal expression of PfGal mRNA in hemocytes challenged by V. alginolyticus was clearly time-dependent and reached the maximum level at 6. h post-challenge, and then recovered to the original level. These results collectively indicated that PfGal may be involved in the immune response against bacterial infection and clearance of bacterial pathogens in P. fucata. © 2011 Elsevier B.V.


Wang Z.,Guangdong Ocean University | Wang Z.,Guangdong Provincial Key Laboratory of Aquaculture in South China Sea for Aquatic Economic Animals | Wang B.,Guangdong Ocean University | Wang B.,Guangdong Provincial Key Laboratory of Pathogenic Biology and Epidemiology for Aquatic Economic Animals | And 7 more authors.
Fish and Shellfish Immunology | Year: 2016

Alpha-2 macroglobulin (α2M) is a ubiquitous protease inhibitor and considered to be an evolutionarily conserved constituent of innate host defence system. Here, an α2M gene (designated as Pfα2M) was obtained from the pearl oyster Pinctada fucata by RT-PCR, PCR walking and rapid amplification of cDNA ends (RACE). The Pfα2M cDNA consists of 6394 bp with an open reading frame (ORF) of 5745 bp encoding a protein of 1914 amino acids with a 19 residues signal peptide. Pfα2M sequence contains three putative functional domains, including a bait region, a thiol ester domain and a receptor-binding domain. Phylogenetic analysis revealed that Pfα2M is closely related to the α2Ms from other molluscs. Pfα2M was expressed in all tested tissues including digestive gland, gill, adductor muscle, mantle and foot, while the highest expression was found in hemocytes. Following challenge with Vibrio alginolyticus, Pfα2M expression in hemocytes was significantly up-regulated at 2 h and then returned to the original level at 48 h. Knockdown of Pfα2M by RNA interference significantly reduced the phagocytosis of V. alginolyticus by hemocytes in vivo, and similar results were obtained upon chemical inactivation of the reactive thioester bond in Pfα2M by methylamine treatment. Taken together, it is suggested that Pfα2M is an immune-relevant molecule and involved in phagocytosis of V. alginolyticus by P. fucata hemocytes, and the function of Pfα2M in phagocytosis is dependent on the active thioester bond. © 2016 Elsevier Ltd

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