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Postoperative care is a critical step of pearl culture that ultimately determines culture success. To determine the effect of dietary vitamin D3 (VD3) levels on immunity and antioxidant capacity of pearl oyster Pinctada fucata martensii during postoperative care and explore the mechanisms behind this phenomenon, five isonitrogenous and isolipidic experimental diets were formulated by adding different levels of dietary VD3 (0, 500, 1000, 3000, and 10000 IU/kg), and the diets were fed to five experimental groups (EG1, EG2, EG3, EG4, and EG5) in turn and cultured indoors. The control group (CG) was cultured in the natural sea. Pearl oysters that were 1.5 years old were subjected to nucleus insertion. After culturing for 30 days, EG3 exhibited significantly higher survival rates than those in CG and EG5 (P 0.05). Moreover, EG3 exhibited the highest activities of alkaline phosphatase, acid phosphatase, catalase, superoxide dismutase, and lysozyme. However, EG5 achieved the highest activities of glutathione peroxidase. Metabolomics-based profiling of pearl oysters fed with high levels of dietary VD3 (EG5) and optimum levels of dietary VD3 (EG3) revealed 76 significantly differential metabolites (SDMs) (VIP 1 and P 0.05). Pathway analysis indicated that SDMs were involved in 21 pathways. Furthermore, integrated key metabolic pathway analysis suggested that pearl oysters in EG5 regulated the pentose phosphate pathway, glutathione metabolism, sphingolipid metabolism, and arachidonic acid metabolism in response to stress generated from excessive VD3. These findings had significant implications on strengthening the future development and application of VD3 in aquaculture of pearl oyster P. f. martensii.

Body color is an important economic and ecological trait in aquatic animals, which influence their economic values and determine the animal ability to survive in the marine environment. Red-colored Plectropomus leopardus is in high demand owing to its bright color and potential value for artificial breeding. High-throughput transcriptome sequencing and liquid chromatography tandem-mass spectrometry (LC-MS/MS) approaches were used to identify transcript and metabolic differences between black-colored and red-colored P. leopardus. Compared with the black-colored group, 218 and 181 differentially expressed genes were up- and down- regulated, respectively, in the red-colored group. 425 and 56 significantly different metabolites were identified in LC-MS positive and LC-MS negative ion models, respectively, between two colored groups. Based on the integrative analysis, the red-colored group exhibited greater carotenoid uptake, transport, and accumulation activity potential than those in the black-colored group and may consume more arachidonic acid for body color formation. The black-colored group showed greater melanin synthesis activity compared with the red-colored group. These results substantially improve the understanding of molecular and metabolic mechanisms underlying body color formation in P. leopardus and the potential of body color for the environment adaptation.

Background Marine bivalves undergo complex development processes, such as shell morphology conversion and changes of anatomy and life habits. In this study, the transcriptomes of pearl oyster Pinctada fucata martensii and Pacific oyster Crassostrea gigas at different development stages were analyzed to determine the key molecular events related to shell formation, settlement and metamorphosis. Result According to the shell matrix proteome, biomineralization-related genes exhibited a consensus expression model with the critical stages of shell formation. Differential expression analysis of P. f. martensii, revealed the negative regulation and feedback of extracellular matrixs as well as growth factor pathways involved in shell formation of larvae, similar to that in C. gigas. Furthermore, neuroendocrine pathways in hormone receptors, neurotransmitters and neuropeptide receptors were involved in shell formation, settlement and metamorphosis. Conclusion Our research demonstrated the main clusters of regulation elements related to shell formation, settlement and metamorphosis. The regulation of shell formation and metamorphosis could be coupled forming the neuroendocrine-biomineralization crosstalk in metamorphosis. These findings could provide new insights into the regulation in bivalve development. Electronic supplementary material The online version of this article (10.1186/s12864-019-5505-8) contains supplementary material, which is available to authorized users.

Glycosaminoglycan (GAG), an important component of proteoglycan (PG), was biosynthesized with the initiation by peptide O-xylosyltransferase. O-xylosyltransferase activity was presented as a marker for increased PG synthesis. In the present study, a novel O-xylosyltransferase (OXT) gene was identified from Pinctada fucata martensii (PmOXT). The PmOXT-deduced protein sequence carried a typical water-soluble carbohydrate domain, branch domain, and xylosyltransferase domain. Homologous analysis of PmOXT presented the conserved DXD motif and catalytic structure characteristics. Phylogenetic tree analysis showed the traditional taxonomy and PmOXT clustered with Crassostrea gigas. PmOXT was expressed in all the detected tissues and developmental stages. PmOXT had a significantly higher expression level in the shell formation associated tissues and developmental stages. PmOXT expressed significantly decreased in the central zone of the mantle and marginal zone of the mantle after RNA interference; additionally, OXT activity and GAG content in the extrapallial fluid were significantly reduced compared with the control. Furthermore, the crystal tablets of prismatic layer displayed obvious holes and disordered crystals with obviously rough surface and irregular crystal tablets were observed in the nacre after RNAi. Results suggested that PmOXT affected the shell formation by influencing the formation of GAGs in the process of addition to the core proteins.

Keratan sulfate possesses considerable amounts of negatively charged sulfonic acid groups and participates in biomineralization. In the present study, we investigated characteristics and functions of a CHST1 gene identified from the pearl oyster Pinctada fucata martensii (PmCHST1b) which participated in the synthesis of keratan sulfate. PmCHST1b amino acid sequence carried a typical sulfotransferase-3 domain (sulfotransfer-3 domain) and belonged to membrane-associated sulfotransferases. Homologous analysis of CHST1 from different species showed the conserved motif (5' PSB motif and 3' PB motif) which interacted with 3'-phosphoadenosine-5'-phosphosulfate (PAPS). Structure analysis of sulfotransferase domain indicted that PmCHST1b showed the conserved catalytic structure character and the relationships presented in the phylogenetic tree conformed to that of traditional taxonomy. Expression pattern of PmCHST1b in different tissues and development stages showed that PmCHST1b widely expressed in all the detected tissues and development stages and showed the highest expression level in the central zone of mantle (MC). PmCHST1b expressed highly in the trochophore, D-stage larvae and spat which corresponded to prodissoconch and dissoconch shell formation, respectively. RNA interference (RNAi) successfully inhibited expression level of PmCHST1b in MC (P

Understanding of nitrogen stable isotope variation in maize hybrids might help obtaining information on nitrogen absorption and distribution in different maize hybrids. In this study, we examined the nitrogen isotopic composition of different parts of maize hybrids under a laboratory culture experiment. The results showed that the δ15N values of different parts of the maize hybrid and its parents were ordered as follows: δ15Nstem>δ15Nleaf>δ15Nroot. The variation pattern of δ15N between the roots and leaves(Δδ15Nroot-leaf) of the maize hybrid was the same as that of δ15N between the roots and stems (Δδ15Nroot-stem). Therefore, the order of Δδ15Nroot-leaf as well as Δδ15Nroot-stem was as follows: Δδ15Nroot-leaf of the maize hybrid>Δδ15Nroot-leaf of the female parent (T4)>Δδ15Nroot-leaf of the male parent (803) and Δδ15Nroot-stem of the maize hybrid>Δδ15Nroot-stem of the female parent (T4)>Δδ15Nroot-stem of the male parent (803). This order is consistent with heterosis, indicating that differences in δ15N reflect the phenomenon of heterosis. The present study provides data in support of using the isotope technique to determine nitrogen distributions inside a plant and guide crossbreeding.

The protein inhibitor of activated STAT (PIAS) functions in diverse aspects, including immune response, cell apoptosis, cell differentiation, and proliferation. In the present study, thePIASin the pearl oysterPinctada fucata martensiiwas characterized. The sequence features of PmPIAS were similar to that of other PIAS sequences with PIAS typical domains, including SAP, Pro-Ile-Asn-Ile-Thr (PINIT), RLD domain, AD, and S/T-rich region. Homologous analysis showed that PmPIAS protein sequence showed the conserved primary structure compared with other species. Ribbon representation of PIAS protein sequences also showed a conserved structure among species, and the PINIT domain and RLD domain showed the conserved structure compared with the sequence ofHomo sapiens. The expression pattern ofPmPIASin different tissues showed significant high expression in the gonad.PmPIASalso exhibited a significantly higher expression in the 1 and 2 days after cold tolerance stress (17°C) and showed its potential in the cold tolerance. The SNP analysis of the exon region ofPmPIASobtained 18 SNPs, and among them, 11 SNPs showed significance among different genotypes and alleles between cold tolerance selection line and base stock, which showed their potential in the breeding for cold tolerance traits.

Fibroblast growth factor 18 (FGF18) plays an important functional role in skeletal growth and development. The FGF18 gene was characterized in pearl oyster Pinctada fucata martensii (PmFGF18) with the full-length sequence containing an open reading frame of 714 bp encoding 237 amino acids. The domain analysis of PmFGF18 showed a distinctive FGF domain, with a high similarity to FGF18 protein sequences from Crassostrea gigas (43.35%) and C. virginica (37.43%). PmFGF18 expression was revealed in all analyzed tissues with a significantly higher expression level in the fast-growing group than the slow-growing group. The analysis of PmFGF18 polymorphism demonstrated 33 SNPs (single nucleotide polymorphisms) in the CDS and promoter region of PmFGF18 sequence. Association analysis revealed 19 SNPs (2 SNPs from CDS and 17 SNPs from the promoter region) associating significantly with growth traits. Among the associated SNPs, one SNP g.50918198 A &gt, C was verified in the other breeding line. Therefore, PmFGF18 can be utilized as a candidate gene for growth, and its related SNPs could be used in selective breeding of P. f. martensii for the improvement of growth traits.

MicroRNAs (miRNAs) are endogenous small non-coding RNAs that regulate gene expression at the post-transcriptional level and play important roles in many biological processes, including controlling body color differentiation in many different animals. However, the mechanism by which specific miRNAs regulate body color formation in leopard coral groupers has remained largely elusive. In the present study, we sequenced the miRNAs of red and black skin samples from leopard coral grouper (Plectropomus leopardus) with the Illumina HiSeq X_Ten platform to explore the function of miRNAs in the body color formation. A total of 15.21 M and 13.69 M high quality reads were obtained from the black and red skin samples, respectively. Within these sequences, 626 known and 354 novel miRNAs were identified. When comparing across the two groups, we found 60 differentially expressed miRNAs, with red-colored group having 44 up-regulated miRNAs including miR-215, miR-2188, miR-194, miR-122, novel-m0118, and 16 down-regulated miRNAs including miR-187, novel-m0189, novel-m0145, compared with black-colored group (p 1). Integrated analysis of differentially expressed miRNAs and mRNAs identified 846 miRNA-mRNA pairs with opposite expression patterns. Go terms and KEGG pathway enrichment analyses showed that these putative target mRNAs were enriched for functions involving in endocrine systems, immune response, and lipid metabolism. The present study shows the potential role of miRNAs in the body color formation of fish and provides a molecular foundation to explain the role of epigenetics in fish body color selections.

Cyclin dependent kinase-7 (Cdk-7) is a protein kinase associated with regulating the cell cycle, cell differentiation and proliferation, apoptosis and inflammatory response. This study characterized the full cDNA sequence of Cdk-7 in Pinctada fucata martensii (PmCdk-7). A full length sequence of 1473bp with an open reading frame (ORF) of 915bp and encodes a 304aa, 5′-UTR of 58bp and a 3′-UTR of 500bp was obtained. The construed amino acid sequence of PmCdk-7 comprised of a Serine/Threonine protein kinases, catalytic (S_TKc) domain with a protein kinases ATP-binding region signature (14-38aa) and the serine/Threonine protein kinases active-site signature (129-141aa) within the domain. Tissue distribution analysis revealed a high relative mRNA expression of PmCdk-7 within haemocytes. Following the insertion operation (grafting), the relative expression levels of PmCdk-7 in the haemocyte was expressed differentially among the studied groups; the black shell colored selected line (BS) and the control group (CG). High expression was recorded between 12 h and 5d with a peak at 3d suggesting a heightened level of DNA replication and inflammatory response during the pearl-sac formation and this expression was higher in BS than CS showcasing, the heightened immune capacity of BS to grafting operation. Immune stimulation experiment with bacterial endotoxin and a viral mimic revealed PmCdk-7 response to pathogenic stress. The results from our study showed that PmCdk-7 performs a vital function during the cell cycle by aiding DNA replication and also aid response to inflammations generated due to the incision from the grafting operation and long exposure to immune-stimulants (pathogens).

The C1q-domain-containing (C1qDC) proteins are a family of proteins characterized by a globular C1q (gC1q) domain in their C-terminus which hold the potential function in the shell formation as shell matrix proteins. In this study, a C1qDC protein was identified and characterized in pearl oyster (Pinctada fucata martensii) (PmC1qDC) to explore its function in nacre formation. The PmC1qDC-deduced protein sequence carried a typical globular C1q (gC1q) domain that possessed the typical 10-stranded β-sandwich fold with a jelly-roll topology common to all C1qDC family members and shared high homology with other gC1q domains. Homologous analysis of PmC1qDC presented it contained conserved secondary structure and Phe135, Phe155, Tyr166, Phe173, Tyr181, Phe183, and Phe256 amino acid residues. Expression pattern analysis showed that PmC1qDC expressed in all the detected tissues and exhibited a significantly higher expression level in nacre formation-associated tissues. After the shell notching, the expression level of PmC1qDC showed significantly up-regulation after 12 h in the central zone of mantle (MC). PmC1qDC expression significantly decreased in the MC after RNA interference (RNAi). Furthermore, disordered crystals with evident rough surface and irregular crystal tablets were observed in the nacre after RNAi. Results suggested that PmC1qDC affects the shell nacre formation, which is significant to improve the pearl production of pearl oyster.

In this study, we developed microsatellite markers, analyzed their distribution and frequency and verified polymorphism based on the transcriptome data of pearl oyster Pinctada maxima. Totally, 58,121 potential EST-SSRs from 145,877 sequences were identified. We randomly selected 100 of these primer pairs for further validation in the stock, 76 primer pairs were successfully amplified and 25 primer pairs were found to be polymorphic. The number of alleles at each locus ranged from 2 to 5, with an average allele number of 2.96. The observed heterozygosity ranged from 0.185 to 0.931 with an average of 0.656. The expected heterozygosity ranged from 0.337 to 0.754 with an average of 0.552. The polymorphic information content ranged from 0.263 to 0.690 with an average of 0.462. These markers will provide an effective tool for genetic diversity, genetic map construction and molecular-assisted breeding in the species of P. maxima. Keywords: Pinctada maxima, SSR markers, Polymorphism, Genetic diversity

Similar to other marine bivalves, Pinctada maxima shows unsynchronized growth, but the underlying metabolic mechanisms have not been studied. In this study, gold-lipped pearl oyster P. maxima from cultured stocks were selected to produce progeny stock. At 180 days, the stock was sorted by size, and fast-growing individuals and slow-growing individuals were separately sampled. Then, a metabolomic approach based on gas chromatography–mass spectroscopy was applied to assess the metabolite changes between the fast-growing and slow-growing groups of P. maxima and to understand the mechanism of their unsynchronized growth. In the metabolomics assay, among the 896 peaks isolated, 111 metabolites were revealed a spectral similarity value of >700 by using mass spectrum matching, and 48 were considered as significantly different metabolites (SDMs; VIP > 1 and P

Natural disasters and environmental pollution are the main problems for traditional offshore culture. Pearl oyster culture through industrial farming can prevent these difficulties, but food availability is limited. In this study, we formulated two diets, P3 and P5, with yeast powder and corn gluten, respectively, as major protein sources. Two experimental groups (EG1 and EG2) were separately fed with P3 and P5 diets indoors. After 45 days of feeding, GC–TOF/MS-based metabolomics of the hepatopancreas was performed to assess the effect of different formulated diets on pearl oyster (Pinctada fucata martensii) combined with growth performance. EG1 displayed significantly faster absolute growth rate and higher survival rate than EG2. In the metabolomic assay, 125 metabolites were identified by mass spectrum matching with a spectral similarity value (SV) > 700, and 35 were considered significantly different metabolites (SDMs) (VIP > 1 and P 0.1). The impact values of phenylalanine, tyrosine, and tryptophan biosynthesis, ascorbate and aldarate metabolism, phenylalanine metabolism, inositol phosphate metabolism, tyrosine metabolism, sulfur metabolism, and cysteine and methionine metabolism were 0.500, 0.500, 0.308, 0.219, 0.207, 0.167, and 0.161, respectively. Further integrated key metabolic pathway analysis showed that EG2 exhibited lower capability for inositol phosphate metabolism, ascorbate and aldarate metabolism, phenylalanine metabolism, and tyrosine metabolism. This study suggested that yeast powder is a better protein source of formulated diet for pearl oysters than corn gluten.

Background: Molluscs can accumulate carotenoids in their body tissues by predominantly feeding on aquatic plant sources. Carotenoid transport and absorption are determined by the regulation of various proteins such as Scavenger receptor class B(SR-BI). We report the identification and characterisation of pearl oyster Pinctada fuctada martensii SR-BI (PmSR-BI). The correlation between total carotenoid content (TCC) and gene expression was also estimated. Results: The full-length cDNA of PmSR-BI was 1828 bp, including an open-reading frame encoding of 1518 bp with a pI value of 5.83. PmSR-BI protein contains a hydrophobic CD36 domain and four centrally clustered cysteine residues for the arrangement of disulphide bridges. The deduced amino acid sequence had an identity of 30% to 60% with the SR-B of other organisms. Reverse transcription polymerase chain reaction analysis showed that mRNA transcripts were expressed in multiple tissues of adult pearl oyster. A higher expression of PmSR-BI gene was observed in the hepatopancreas than in the adductor muscle, gill and mantle. The TCC and gene expression of PmSR-BI were significantly correlated (P

In this study, we formulated five diets, namely, P1, P2, P3, P4 and P5, with Chlorella sp. powder, Spirulina platensis powder, yeast powder, soybean meal and corn gluten, respectively, as major protein sources. A feeding experiment was designed to evaluate the effects of formulated diets on the growth performance, immunity and antioxidant and biomineralization capacity of juvenile pearl oyster (Pinctada fucata martensii). In the experiments, the five groups were separately fed with P1, P2, P3, P4 and P5 diets. After 45 days of feeding, pearl oysters fed on P1, P2, P3 and P4 diets showed significantly higher absolute growth rate and protease and amylase activities than those fed on P5 diet (P < 0.05). Moreover, pearl oysters fed on P1, P2, P3 and P4 diets exhibited significantly higher activities of alkaline phosphatase (AKP), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) (P < 0.05). Significantly higher expression levels of SOD, GPx, CAT, heat shock protein (HSP) 70, HSP90, nacrein, pif177 and pearlin mRNA were observed in pearl oysters fed on P1, P2, P3 and P4 diets relative to those fed on P5 (P < 0.05). Results suggested the suitability of Chlorella sp. powder, S. platensis powder, yeast powder and soybean meal as protein sources for development of formulated diets for pearl oyster P. f. martensii.

Background : C4ST-1 catalyses the transfer of sulfate groups in the sulfonation of chondroitin during chondroitin sulfate synthesis. Chondroitin sulfate consists of numerous copies of negatively charged sulfonic acid groups that participate in the nucleation process of biomineralization. In the present study, we obtained two CHST11 genes ( PmCHST11a and PmCHST11b ) which encoded the C4ST-1 and explored the functions of these genes in the synthesis of chondroitin sulfate and in the formation of the nacreous layer of shells. Results: Both PmCHST11a and PmCHST11b had a sulfotransferase-2 domain, a signal peptide and a transmembrane domain. These properties indicated that these genes localise in the Golgi apparatus. Real-time PCR revealed that both PmCHST11a and PmCHST11b were highly expressed in the central zone of the mantle tissue. Inhibiting PmCHST11a and PmCHST11b via RNA interference significantly decreased the expression levels of these genes in the central zone of the mantle tissue and the concentration of chondroitin sulfate in extrapallial fluid. Moreover, shell nacre crystallised irregularly with a rough surface after RNA interference. Conclusions: This study indicated that PmCHST11a and PmCHST11b are involved in the nacre formation of P. fucata martensii through participating in the synthesis of chondroitin sulfate. Normal 0 false false false EN-US ZH-CN X-NONE /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Tabla normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-parent:""; mso-padding-alt:0cm 5.4pt 0cm 5.4pt; mso-para-margin:0cm; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:10.5pt; mso-bidi-font-size:11.0pt; font-family:"Calibri",sans-serif; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin; mso-bidi-font-family:"Times New Roman"; mso-bidi-theme-font:minor-bidi; mso-font-kerning:1.0pt; mso-fareast-language:ZH-CN;}

The pearl oyster, Pinctada fucata martensii, produces high-quality pearls. During pearl production, excess immune and inflammatory response after transplantation will lead to nucleus rejection, pearl sac formation failure, and death of the host pearl oyster. The hemocyte transcriptome and fatty acid (FA) contents in the adductor muscle before and after transplantation were analyzed to investigate the response of pearl oyster P. f. martensii to allograft-induced stress from lipid metabolism. The hemocyte transcriptome analysis detected 193 lipid metabolism-related genes, such as the elongation of very long-chain FA protein 5, acyl-CoA 6-desaturase, cytochrome P450, phospholipase A2, glycerol-3-phosphate O-acyltransferase, and prostaglandin-H2 d-isomerase. Pathway enrichment analyses revealed that these genes were mainly involved in the "biosynthesis of unsaturated FAs," "FA biosynthesis," "ARA metabolism," and "glycerolipid metabolism." An analysis of FA contents in the adductor muscle indicated no significant difference in the contents of lauric acid, myristic acid, pentadecanoic acid, palmitic acid, palmitoleic acid, heptadecanoic acid, stearic acid, oleic acid, vaccenic acid, linoleic acid, arachidic acid, α-linolenic acid, eicosadienoic acid, docosadienoic acid, and 11,14,17-eicosatrienoic acid. However, ARA, DHA, and EPA in the adductor muscle after transplantation were significantly greater than those processed without grafting surgery. These results suggest that pearl oysters require more polyunsaturated FAs (PUFAs) to regulate their inflammatory and immune response after transplantation. However, their ability to biosynthesize unsaturated FAs is limited. Given these results, the addition of PUFA-containing diets or selection of a line with strong ability to biosynthesize unsaturated FAs may be valuable for pearl oyster recovery after transplantation.

Larval development, especially settlement and metamorphosis, has a vital influence on commercial culture during the life cycle of marine invertebrates. Sipunculus nudus, a peanut worm, is an important economic aquaculture species. However, information about the mechanism of its larval settlement and metamorphosis is still limited. Transcriptomic analysis was performed to determine the DEGs between the pelagospheric larva and creeping larva of S. nudus. A total of 96,455 unique unigenes were obtained and 46,355 were annotated. Transcriptome analysis revealed 19,352 DEGs, including 9,923 that were upregulated and 9,429 downregulated after settlement and metamorphosis. KEGG pathway enrichment analyses revealed that these DEGs were mainly involved in the Notch signaling pathway, TNF signaling pathway, protein digestion and absorption, thyroid hormone signaling pathway, and ECM–receptor interaction, and so on. Furthermore, the changes in genes related to cytoskeleton and cell adhesion indicated that protein degradation and apoptosis during metamorphosis mediate the loss of larval cilia and ECM remodeling functions in S. nudus metamorphosis. Changes in genes related to signal transduction and nervous system suggested the cessation of larval swimming, which is involved in settlement and metamorphosis. Changes in genes related to ingestion and digestion indicated that dietary shift occurred with metamorphic transition. Changes in some genes related to stress response and immunity indicated that conditioning induced the immune response of peanut worm. The results improve understanding of the physiological traits controlling S. nudus metamorphosis and provide a solid basis for further study.

Odd-skipped related 1 (OSR1) from Pinctada fucata martensii (PmOSR1) was cloned in this study. PmOSR1 contained an open reading frame measuring 951 bp that encoded a polypeptide of 316 residues. Domain analysis also revealed that PmOSR1 had a typical ZnF_C2H2 domain, indicating high identity with OSR1 protein sequence from Crassostrea gigas (55.25%) and Mizuhopecten yessoensis (65.48%). The expression pattern analysis of PmOSR1 revealed its wide expression in the tissues and developmental stages. PmOSR1 also presented a higher expression in the fast-growing group and hybrid families than in the slow-growing group and inbred families. The polymorphism analysis of PmOSR1 revealed 45 single nucleotide polymorphisms (SNPs) in the genomic sequence, and association analysis presented four SNPs significantly associated with growth traits (namely, shell length, width, height, and shell weight; P

Chitin participates in shell formation as the main component of an organic framework. Chitin-binding protein contains domains that can bind to chitin specifically. In this study, a novel chitin-binding protein from Pinctada fucata martensii (PmCBP) with poly (chitin-binding domain) was cloned, which contains a 5′-untranslated region (UTR) of 114 bp and 3′UTR of 116 bp, and encodes a putative protein of 2044 amino acids. The predicted PmCBP protein was structurally typical of the CBP family with 20 ChtBD2 domains. Phylogenetic and linear relation analyses showed that the ChtBD2 domain has a highly conserved structure among the three species of P. f. martensii, Crassostrea gigas, and Mizuhopecten yessoensis. qRT-PCR and in-situ hybridization analysis revealed that PmCBP was most abundant in the mantle pallium whose expression level was significantly correlated with the growth traits. After RNAi, PmCBP expression was significantly inhibited in the mantle pallium (P

Similar to other marine bivalves, Pinctada fucata martensii presents unsynchronized growth, which is one of the problems farmers currently face. However, the underlying mechanisms have not been studied. In the present study, pearl oyster P. f. martensii from cultured stocks were selected to produce a progeny stock. At 180 days, the stock was sorted by size, and fast- and slow-growing individuals were separately sampled. Then, metabolomic and transcriptomic approaches were applied to assess the metabolic and transcript changes between the fast- and slow-growing P. f. martensii groups and understand the mechanism underlying their unsynchronized growth. In the metabolomics assay, 30 metabolites were considered significantly different metabolites (SDMs) between the fast- and slow-growing groups and pathway analysis indicated that these SDMs were involved in 20 pathways, including glutathione metabolism; sulfur metabolism; valine, leucine, and isoleucine biosynthesis; and tryptophan metabolism. The transcriptome analysis of different growth groups showed 168 differentially expressed genes (DEGs) and pathway enrichment analysis indicated that DEGs were involved in extracellular matrix-receptor interaction, pentose phosphate pathway, aromatic compound degradation. Integrated transcriptome and metabolome analyses showed that fast-growing individuals exhibited higher biomineralization activity than the slow-growing group, which consumed more energy than the fast-growing group in response to environmental stress. Fast-growing group also exhibited higher digestion, anabolic ability, and osmotic regulation ability than the slow-growing group. This study is the first work involving the integrated metabolomic and transcriptomic analyses to identify the key pathways to understand the molecular and metabolic mechanisms underlying unsynchronized bivalve growth.

Land-based culturing can avoid the effects of environmental pollution and natural disasters, thus ensuring food safety for shellfish. However, food availability, in this case, is limited. To achieve the optimum balance of dietary carbohydrates and proteins and explore the mechanisms behind the phenomenon, we formulated five isoenergetic and isolipidic diets (C30P40, C35P35, C40P30, C45P25, and C50P20) with different levels of carbohydrates (C) and proteins (P). There were five experimental groups (C30P40, C35P35, C40P30, C45P25, and C50P20) and two control groups (CG1 and CG2). CG1 was fed with mixed powders of yeast and Chlorella sp., and CG2 was cultured in natural sea. After 60-day feeding, the highest rates of survival and absolute growth appeared in C45P25. C45P25 exhibited significantly higher activities of amylase, protease, alkaline phosphatase, acid phosphatase, superoxide dismutase, catalase, glutathione peroxidase, and phenoloxidase and significantly lower malondialdehyde content than C30P40, C35P35, C40P30, C50P20, and CG1. No significant differences were observed between C45P25 and CG2. Furthermore, the total antioxidant capacity of the pearl oysters in C45P25 was significantly higher than that in C30P40, C35P35, C40P30, and C50P20. On the basis of these results, the optimal balance of proteins and carbohydrates for pearl oysters was the C45P25 diet. Metabolomics-based profiling of the pearl oysters fed with high-carbohydrate/low-protein diet (C45P25) and low-carbohydrate/high-protein diet (C30P40) revealed 80 significantly different metabolites (VIP 1 and P 0.05). Furthermore, integrated key metabolic pathway analysis showed that C45P25 regulated starch and sucrose metabolism, alanine, aspartate and glutamate metabolism and glycine, serine and threonine metabolism to meet the energy demand and increase the glucogenic amino acid, thereby promoting protein synthesis and reducing fatty acid β-oxidation in comparison with C30P40. This finding helps elucidate the underlying mechanisms leading to the high-carbohydrate/low-protein diet characteristic of the optimal dietary carbohydrate and protein levels of P. f. martensii.

Natural disasters and environmental pollution are the main problems in traditional offshore cultivation. While culturing pearl oysters through industrial farming can avoid these problems, food availability in this case is limited. This study compares the metabolomics responses of pearl oysters, Pinctada fucata martensii, fed a formulated diet indoors with those of oysters cultured with natural diet outdoors by using a gas chromatography time-of-flight mass spectrometry (GC-TOF/MS)-based metabolomics approach. The animals were divided into two groups as follows: the experimental group (EG) was fed a formulated diet indoors and the control group (CG) was cultured with natural diet outdoors. After 45 days of feeding, the survival rate of EG was significantly higher than that of CG. The absolute growth rate (AGR) of the total weight of EG did not significantly differ from that of CG, but the AGRs of the shell length, shell height, and shell width of CG were significantly higher than those of EG. EG showed significantly higher amylase activities than CG, and the hexokinase and glucose-6-phosphate isomerase concentrations of the former were significantly lower than those of the latter. Metabolomics revealed 125 metabolites via mass spectrum matching with a spectral similarity value > 700 in the hepatopancreas, and 48 metabolites were considered to be significantly different between groups (VIP > 1 and P < 0.05). Pathway analysis results indicated that these significantly different metabolites were involved in 34 pathways. Further integrated key metabolic pathway analysis showed that, compared with CG, EG had lower capabilities for cysteine and methionine metabolism, sulfur metabolism, and starch and sucrose metabolism. This study demonstrated that the formulated diet could be an excellent substitute for natural diet; however, its nutrients were insufficient. Effective strategies should be developed to enhance the utilization of formulated diets.

Marine pearl production is directly influenced by the growth speed of Pinctada fucata martensii. However, the slow growth rate of this organism remains the main challenge in aquaculture production. Epidermal growth factor receptor (EGFR), an important receptor of tyrosine kinases in animals, plays versatile functions in development, growth and tissue regeneration. In this study, we described the characteristic and function of an EGFR gene identified from P. f. martensii (PmEGFR). PmEGFR possesses a typical EGFR structure and is expressed in all studied tissues, with the highest expression level in adductor muscle. PmEGFR expression level is significantly higher in the fast-growing group than that in the slow-growing one. Correlation analysis represents that shell height and shell weight show positive correlation with PmEGFR expression (p p PmEGFR is a valuable functional gene associated with growth traits. PmEGFR expression level is positively correlative with growth traits of P. f. martensii, which indicates PmEGFR is a valuable functional gene associated with growth traits.

Nacre, the iridescent material found in pearls and shells of molluscs, is formed through an extraordinary process of matrix-assisted biomineralization. Despite recent advances, many aspects of the biomineralization process and its evolutionary origin remain unknown. The pearl oyster Pinctada fucata martensii is a well-known master of biomineralization, but the molecular mechanisms that underlie its production of shells and pearls are not fully understood. We sequenced the highly polymorphic genome of the pearl oyster and conducted multi-omic and biochemical studies to probe nacre formation. We identified a large set of novel proteins participating in matrix-framework formation, many in expanded families, including components similar to that found in vertebrate bones such as collagen-related VWA-containing proteins, chondroitin sulfotransferases, and regulatory elements. Considering that there are only collagen-based matrices in vertebrate bones and chitin-based matrices in most invertebrate skeletons, the presence of both chitin and elements of collagen-based matrices in nacre suggests that elements of chitin- and collagen-based matrices have deep roots and might be part of an ancient biomineralizing matrix. Our results expand the current shell matrix-framework model and provide new insights into the evolution of diverse biomineralization systems.

Highlights • This study obtains the full length of Pmtgfbr1 of the pearl oyster P. fucata martensii. • Pmtgfbr1 possesses the conserved domain of Tgfbr1. • Pmtgfbr1 holds negatively effect on the growth of P. fucata martensii., Pinctada fucata martensii is cultured for pearl production. Growth improvement has received considerable research interest. Transforming growth factor β type Ⅰ receptor (TβR-I), which is involved in signals transmission of transforming growth factor beta (TGF-β), participates in cell proliferation and growth. In this study, we characterized a Tgfbr1 gene which encoded TβR-I from P. fucata martensii (Pmtgfbr1). Pmtgfbr1 cDNA contains an open reading frame of 1569 bp and encodes a polypeptide of 522 amino acids (aa). Pmtgfbr1 possesses a typical TβR-I structure (extracellular receptor ligand domain, transmembrane domain, and cytoplasmic tyrosine kinase catalytic domain). Pmtgfbr1 is expressed in all the studied tissues and exhibited the highest expression level in the adductor muscle. Moreover, Pmtgfbr1 exhibited the lower expression level in the larger group (L) than that in the smaller group (S) and is negatively correlated with growth traits (P