Scallop Patinopecten Yessoensis Research Articles

The involvement of CaMKKI in activating AMPKα in yesso scallop Patinopecten yessoensis under high temperature stress

Calcium/calmodulin dependent protein kinase kinase (CaMKK), a highly conserved protein kinase, is involved in the downstream processes of various biological activities by phosphorylating and activating 5′-AMP-activated protein kinase (AMPK) in response to the increase of cytosolic-free calcium (Ca2+). In the present study, a CaMKKI was identified from Yesso scallop Patinopecten yessoensis. Its mRNA was ubiquitously expressed in haemocytes and all tested tissues with the highest expression level in mantle. The expression level of PyCaMKKI mRNA in adductor muscle was significantly upregulated at 1, 3 and 6 h after high temperature treatment (25 °C), which was 3.43-fold (p < 0.05), 5.25-fold (p < 0.05), and 5.70-fold (p < 0.05) of that in blank group, respectively. At 3 h after high temperature treatment (25 °C), the protein level of PyAMPKα, as well as the phosphorylation level of PyAMPKα at Thr170 in adductor muscle, and the positive co-localized fluorescence signals of PyCaMKKI and PyAMPKα in haemocyte all increased significantly (p < 0.05) compared to blank group (18 °C). The pull-down assay showed that rPyCaMKKI and rPyAMPKα could bind each other in vitro. After PyCaMKKI was silenced by siRNA, the mRNA and protein levels of PyCaMKKI and PyAMPKα, and the phosphorylation level of PyAMPKα at Thr170 in adductor muscle were significantly down-regulated (p < 0.05) compared with the negative control group receiving an injection of siRNA-NC. These results collectively suggested that PyCaMKKI was involved in the activation of PyAMPKα in response to high temperature stress and would be helpful for understanding the function of PyCaMKKI-PyAMPKα pathway in maintaining energy homeostasis under high temperature stress in scallops.

The involvement of CaMKKI in activating AMPKα in Yesso scallop Patinopecten yessoensis under high temperature stress

Calcium/calmodulin dependent protein kinase kinase (CaMKK), a highly conserved protein kinase, is involved in the downstream processes of various biological activities by phosphorylating and activating 5'-AMP-activated protein kinase (AMPK) in response to the increase of cytosolic-free calcium (Ca2+). In the present study, a CaMKKI was identified from Yesso scallop Patinopecten yessoensis. Its mRNA was ubiquitously expressed in haemocytes and all tested tissues with the highest expression level in mantle. The expression level of PyCaMKKI mRNA in adductor muscle was significantly upregulated at 1, 3 and 6 h after high temperature treatment (25 °C), which was 3.43-fold (p < 0.05), 5.25-fold (p < 0.05), and 5.70-fold (p < 0.05) of that in blank group, respectively. At 3 h after high temperature treatment (25 °C), the protein level of PyAMPKα, as well as the phosphorylation level of PyAMPKα at Thr170 in adductor muscle, and the positive co-localized fluorescence signals of PyCaMKKI and PyAMPKα in haemocyte all increased significantly (p < 0.05) compared to blank group (18 °C). The pull-down assay showed that rPyCaMKKI and rPyAMPKα could bind each other in vitro. After PyCaMKKI was silenced by siRNA, the mRNA and protein levels of PyCaMKKI and PyAMPKα, and the phosphorylation level of PyAMPKα at Thr170 in adductor muscle were significantly down-regulated (p < 0.05) compared with the negative control group receiving an injection of siRNA-NC. These results collectively suggested that PyCaMKKI was involved in the activation of PyAMPKα in response to high temperature stress and would be helpful for understanding the function of PyCaMKKI-PyAMPKα pathway in maintaining energy homeostasis under high temperature stress in scallops.

Inhibition of TRPA1-like alleviated unfolded protein response and apoptosis by regulating cytoplasmic Ca2+ in Yesso scallop Patinopecten yessoensis under high temperature stress

Transient receptor potential ankyrin subtype 1 (TRPA1) is a nonselective cation channel protein typically forms ion channels that regulate intracellular calcium homeostasis, and can be induced by temperature and various chemicals. In the present study, the involvement of PyTRPA1-like in regulating unfolded protein response (UPR) and apoptosis in Yesso Scallop Patinopecten yessoensis under high temperature stress was investigated. The mRNA transcripts of PyTRPA1-like were detected in haemocytes and all the examined tissues with the highest expression level in mantle. After TRPA1 activator (allyl-isothiocyanate, AITC) and high temperature (25°C) treatment, the expression level of PyTRPA1-like mRNA and the Ca2+ content in haemocytes increased significantly (p &amp;lt; 0.05) at 3 h, and then recovered to the normal level at 12 h, and the expression level of PyGRP78, PyIRE1, PyATF6β, PyPERK and PyCaspase-3 mRNA in haemocytes, and Caspase-3 activity and apoptosis rate were also significantly upregulated (p &amp;lt; 0.05). After TRPA1 antagonist (HC-030031) and high temperature (25°C) treatment, the intracellular Ca2+ content, the transcripts of PyGRP78, PyIRE1 and PyCaspase-3 in haemocytes, as well as the Caspase-3 activity and apoptosis rate decreased significantly compared to the control group (p &amp;lt; 0.05), while the Ca2+ distribution in haemocytes showed no difference with that in control group. These results collectively suggest that PyTRPA1-like plays important roles in regulating UPR and apoptosis by mediating calcium influx under high temperature stress in scallop P. yessoensis.

Cryopreservation of Yesso scallop (Patinopecten yessoensis) sperm

The germplasm of farmed Yesso scallop Patinopecten yessoensis in China has deteriorated since its introduction more than 40 years ago. The main aim of this study is to develop a sperm cryopreservation technique to assist the newly established program to manage this issue in China. This study investigated the factors important to the development of a non-programmable sperm cryopreservation technique in this species, including cryoprotectant agent [CPA; dimethyl sulfoxide (DMSO), propylene glycol (PG), ethylene glycol (EG), glycine, sucrose, glucose and trehalose], equilibration time, rack height and thawing temperature. A low post-thaw sperm fertilization rate of 27.67 ± 2.52% was produced with the parameters optimized with the permeable CPAs (DMSO, PG and EG) only: 6% DMSO, 10 min equilibration time, 5 cm rack height and 30 °C thawing temperature. This rate was further improved to 44.00 ± 2.00% by adding 3% glucose into 6% DMSO. This addition had also improved the integrities of post-thaw sperm DNA, plasma membrane and acrosome, mitochondrial membrane potential and activities of some enzymes. Results from this study also showed that the post-thaw sperm morphology and ultrastructure were intensively compromised. In addition, the sperm agglutination found in the newly spawned sperm might be one of the phenomena resulted from the so-called germplasm deterioration, especially in the stock used in this study. The further exacerbation of agglutination after cryopreservation would also compromise the post-thaw sperm performance. The sperm cryopreservation technique established in this study would provide a better option to assist in managing the genetic diversity of farmed Yesso scallops in China.

Paralytic shellfish toxins producing dinoflagellates cause dysbacteriosis in scallop gut microbial biofilms

Filter-feeding bivalves could accumulate paralytic shellfish toxins (PSTs) produced by harmful dinoflagellates through diet. Despite that bivalves are resistant to these neurotoxins due to possessing PST-resistant sodium channel, exposure to PSTs-producing dinoflagellates impair bivalve survival. We hypothesized that ingesting PSTs-producing dinoflagellates may influence the gut microbiota, and then the health of bivalves. To test this idea, we compared the gut microbiota of the scallop Patinopecten yessoensis, after feeding with PST-producing or non-toxic dinoflagellates. Exposure to PSTs-producing dinoflagellates resulted in a decline of gut microbial diversity and a disturbance of community structure, accompanied by a significant increase in the abundance and richness of pathogenic bacteria, represented by Vibrio. Moreover, network analysis demonstrated extensive positive correlations between pathogenic bacteria abundances and PSTs concentrations in the digestive glands of the scallops. Furthermore, isolation of a dominant Vibrio strain and its genomic analysis revealed a variety of virulence factors, including the tolC outer membrane exporter, which were expressed in the gut microbiota. Finally, the infection experiment demonstrated scallop mortality caused by the isolated Vibrio strain; further, the pathogenicity of this Vibrio strain was attenuated by a mutation in the tolC gene. Together, these findings demonstrated that the PSTs may affect gut microbiota via direct and taxa-specific interactions with opportunistic pathogens, which proliferate after transition from seawater to the gut environment. The present study has revealed novel mechanisms towards deciphering the puzzles in environmental disturbances-caused death of an important aquaculture species.

PyLKB1 regulates glucose transport via activating PyAMPKα in Yesso Scallop Patinopecten yessoensis under high temperature stress

Liver kinase B1 (LKB1) is a classical serine/threonine protein kinase and plays an important role in maintaining energy homeostasis through phosphorylate AMP-activated protein kinase α subunit (AMPKα). In this study, a homologous molecule of LKB1 with a typical serine/threonine kinase domain and two nuclear localization sequences (NLSs) was identified in Yesso Scallop Patinopecten yessoensis (PyLKB1). The mRNA transcripts of PyLKB1 were found to be expressed in haemocytes and all the examined tissues, including gill, mantle, gonad, adductor muscle and hepatopancreas, with the highest expression level in hepatopancreas. PyLKB1 was mainly located in cytoplasm and nucleus of scallop haemocytes. At 3 h after high temperature stress treatment (25 °C), the mRNA transcripts of PyLKB1, PyAMPKα, and PyGLUT1 in hepatopancreas, the phosphorylation level of PyAMPKα at Thr170 in hepatopancreas, the positive fluorescence signals of PyLKB1 in haemocytes, glucose analogue 2-NBDG content in haemocytes, and glucose content in hepatopancreas, haemocytes and serum all increased significantly (p < 0.05) compared to blank group (15 °C). However, there was no significant difference at the protein level of PyLKB1 and PyAMPKα. After PyLKB1 was knockdown by siRNA, the mRNA expression level of PyGLUT1, and the glucose content in hepatopancreas and serum were significantly down-regulated (p < 0.05) compared with the negative control group receiving an injection of siRNA-NC. However, there were no significant difference in PyGLUT1 expression, glucose content and glucose analogue 2-NBDG content in haemocytes. These results collectively suggested that PyLKB1-PyAMPKα pathway was activated to promote glucose transport by regulating PyGLUT1 in response to high temperature stress. These results would be helpful for understanding the function of PyLKB1-PyAMPKα pathway in regulating glucose metabolism and maintaining energy homeostasis under high temperature stress in scallops.

Monovalent Salt and pH-Stimulated Gelation of Scallop (Patinopecten yessoensis) Male Gonad Hydrolysates/κ-Carrageenan.

The gelation of scallop Patinopecten yessoensis male gonad hydrolysates (SMGHs) and κ-carrageenan (KC) subjected to pH (2-8, 3-9) and NaCl/KCl stimuli-response was investigated. SMGHs/KC gels subjected to a NaCl response exhibited an increasing storage modulus G'from 2028.6 to 3418.4 Pa as the pH decreased from pH 8 to 2, with corresponding T23 fluctuating from 966.40 to 365.64 ms. For the KCl-treated group, SMGHs/KC gels showed an even greater G' from 4646.7 to 10996.5 Pa, with T23 fluctuating from 622.2 to 276.98 ms as the pH decreased from 9 to 3. The improved gel strength could be ascribed to the blueshift and redshift of hydroxyl groups and amide I peaks, enhanced enthalpy and peak temperature, and gathered characteristic diffraction peaks from SMGHs, KC, NaCl, and KCl. The CLSM and cryo-SEM images further reflected that SMGHs/KC gels showed more flocculation formation and denser and more homogeneous networks with smaller pore sizes in more acidic domains, especially when subjected to the KCl response. This research gives a theoretical and methodological understanding of the construction of salt- and pH-responsive SMGHs/KC hydrogels as novel functional soft biomaterials applied in food and biological fields.

Ontogenetic Changes during the Life Span of the Scallop Patinopecten yessoensis Determined Using a Geometric Morphometric Method

Ontogenetic Changes during the Life Span of the Scallop Patinopecten yessoensis Determined Using a Geometric Morphometric Method

Formation and microstructural characterization of scallop (Patinopecten yessoensis) male gonad hydrolysates/sodium alginate coacervations as a function of pH

Studying the noncovalent interactions between proteins and polysaccharides is quite important mainly due to the wide number of applications such as developing pH-responsive complexes. Scallop Patinopecten yessoensis male gonad hydrolysates‑sodium alginate (SMGHs-SA) was investigated as noncovalent complexes at pH from 1 to 10. The critical pH values pHC (around 6) and pHφ (around 4) were independent of the SMGHs-SA ratio, indicating the formation of soluble and insoluble complexes. The pH response of SMGHs-SA complexes was evaluated by investigating the rheological behavior, moisture distribution, functional group change and microstructure. Compared to the co-soluble and soluble complexes phases, the SMGHs-SA complexes had a higher storage modulus and viscosity as well as a lower relaxation time (T23) in the insoluble complexes phase (pHφ>3). Additionally, the amide I band and COO− stretching vibration peaks were redshifted and the amide A band vibration peaks were blueshifted by acidification. Electrostatic interactions and intermolecular/intramolecular hydrogen bonding led to SMGHs-SA agglomeration at pH 3, forming a uniform and dense gel network structure with strong gel strength and water-retention capacity. This study provides a theoretical and methodological basis for the design of novel pH-responsive complexes by studying SMGHs-SA complex coacervation.

The intestinal bacterial community over seasons and its relationship with physiological status of Yesso scallop Patinopecten yessoensis

Emerging evidence indicates that the intestinal bacterial communities associated with eukaryotes play critical roles in the physiological activities and health of their hosts. Yesso scallop Patinopecten yessoensis, one of the cold-water aquaculture species in the North Yellow Sea of China, has suffered from massive mortality in recent years. In the present study, P. yessoensis were collected from Zhangzi Island, Dalian from March 2021 to January 2022 to investigate the intestinal bacterial community and physiological indices. 16S rRNA gene sequencing data revealed that the diversity of intestinal bacteria changed significantly over seasons, with the highest Chao1 (237.42) and Shannon (6.13) indices detected in January and the lowest Chao1 (115.44) and Shannon (2.73) indices detected in July. Tenericutes, Proteobacteria and Firmicutes were dominant phyla in the intestinal bacteria of P. yessoensis, among which Firmicutes and Proteobacteria significantly enriched in August and January, respectively. Mycoplasma was the most abundant genus during the sampling period, which exhibited the highest abundance in October (75.26%) and lowest abundance in August (13.15%). The functional profiles of intestinal bacteria also exhibited seasonal variation, with the pathways related to pentose phosphate and deoxyribonucleotides biosynthesis enriched in August while the glycogen biosynthesis pathway enriched in October. Redundancy analysis showed that seawater pH, dissolved inorganic nitrogen and silicate were major environmental factors driving the temporal succession of scallop intestinal bacteria. Correlation clustering analysis suggested that the relative abundances of Endozoicomonas and Vibrio in the intestine were positively correlated with superoxide dismutase activity in hepatopancreas while negatively correlated with malondialdehyde content in hepatopancreas and glycogen content in adductor muscle. All the results revealed that the intestine harbored a lower bacterial diversity and a higher abundance of Vibrio in August, compared to January, which were closely related to the oxidative stress status of scallop in summer. These findings will advance our understanding of the relationship between seasonal alteration in the intestinal bacteria and the physiological status of scallops.

The involvement of AMP-activated protein kinase α in regulating glycolysis in Yesso scallop Patinopecten yessoensis under high temperature stress

AMP-activated protein kinase α subunit (AMPKα), the central regulatory molecule of energy metabolism, plays an important role in maintaining energy homeostasis and helping cells to resist the influence of various adverse factors. In the present study, an AMPKα was identified from Yesso scallop Patinopecten yessoensis (PyAMPKα). The open reading frame (ORF) of PyAMPKα was of 1599 bp encoding a putative polypeptide of 533 amino acid residues with a typical KD domain, a α-AID domain and a α-CTD domain. The deduced amino acid sequence of PyAMPKα shared 59.89–74.78% identities with AMPKαs from other species. The mRNA transcripts of PyAMPKα were found to be expressed in haemocytes and all the examined tissues, including gill, mantle, gonad, adductor muscle and hepatopancreas, with the highest expression level in adductor muscle. PyAMPKα was mainly located in cytoplasm of scallop haemocytes. At 3 h after high temperature stress treatment (25 °C), the mRNA transcripts of PyAMPKα, the phosphorylation level of PyAMPKα at Thr170 and the lactic acid (LD) content in adductor muscle all increased significantly, while the glycogen content decreased significantly. The activity of pyruvate kinase (PyPK) and the relative mRNA expression level of phosphofructokinase (PyPFK) were significantly up-regulated at 3 h after high temperature stress treatment (25 °C). Furthermore, the PyAMPKα activator AICAR could effectively upregulate the phosphorylation level of PyAMPKα, and increase activities of PyPFK and pyruvate kinase (PyPK). Meanwhile the glycogen content also declined under AICAR treatment. These results collectively suggested that PyAMPKα was involved in the high temperature stress response of scallops by enhancing glycolysis pathway of glycogen. These results would be helpful for understanding the functions of PyAMPKα in maintaining energy homeostasis under high temperature stress in scallops.

Study on the Regulation Mechanism of Quality Deterioration Due to Chilling Stress and Dry Exposure during Anhydrous Storage and Transportation of Yesso Scallop Patinopecten yessoensis.

In this paper, the quality change of Yesso scallop (Patinopecten yessoensis) in the process of anhydrous storage and transportation after cold acclimation and induced dormancy was studied, and the regulation mechanism of quality degradation during storage and transportation in the process of gradient chilling stress and drying exposure was further explored. The results show that, when transferred from hydrous to anhydrous states, the breathing pattern of the scallops changed from aerobic to anaerobic. Their gill filaments were altered and their apparent vitality constantly declined, which was reflected by the edge shrinkage of the pallium and the direct proportions of the edge reduction rate and the stimulus response period. After being in the anhydrous state for 4 d, the AEC value dropped to 67.59%. At this time, if they were placed under hydration again, the scallops resumed a good growth state. By proteomics analysis, it was revealed that cold acclimation and dry exposure mainly led to changes in biological functions and pathways, such as mitochondrial inner membrane and ATP hydrolysis activity. In addition, it can be seen from the functional annotation and enrichment analysis of the metabolite KEGG that cold acclimation promoted the purine metabolism of scallops, while dry exposure inhibited the metabolism of saturated fatty acids. In this study, the infrared sensing mode was used for the first time, too, in order to record the heart-rate changes of the scallops during circulation, which shows that non-destructive vitality monitoring of Lamellibranchia is feasible.

Effect of high temperature stress on glycogen metabolism in gills of Yesso scallop Patinopecten yessoensis.

Glycogen is the main energy storage material in mollusc, and the regulation of its metabolism is essential for the response against high temperature stress. In the present study, the alternation of lactic acid (LD) content, glycogen reserves, mRNA expression level of genes encoding glycogen metabolism enzymes and activities of glycogen metabolism enzymes in gills of Yesso scallop Patinopecten yessoensis after an acute high temperature treatment at 25°C were examined to understand the effect of high temperature on glycogen metabolism. The activity of T-ATPase in gills of scallops presented a gradual increase trend especially at 6h after the acute high temperature treatment (p<0.05). The glycogen reserves did not change significantly even there was a downward trend at 24h after the acute high temperature treatment (p>0.05). The mRNA transcripts of glycogen synthase (PyGCS) in gills of scallops decreased significantly at 1, 3, 6 and 12h (p<0.05), and recovered to normal level at 24h (p>0.05) after the acute high temperature treatment, while that of glycogen phosphorylase a (PyGPa) and phosphoenol pyruvate carboxy kinase (PyPEPCK) were both significantly down-regulated from 1h to 24h (p<0.05) after the acute high temperature treatment. The activity of PyGPa at 1, 12 and 24h and the content of LD at 3 and 24h in gills of scallops after the acute high temperature treatment both increased significantly (p<0.05). Furthermore, the mRNA transcripts of hexokinase (PyHK) and pyruvate kinase (PyPK) in gills of scallops increased significantly (p<0.05) after the acute high temperature treatment, and the response of PyHK was stronger. However, there was no significant difference on the activity of PyPK in gills of scallops between the experimental samples and the blank samples (p>0.05). In addition, the mRNA transcripts of citrate synthase (PyCS) in gills of scallops were significantly down-regulated at 6h and 12h (p<0.05), and finally returned to normal level at 24h (p>0.05) after the acute high temperature treatment. These results collectively indicated acute high temperature stress leaded the alternation of glycogen metabolism in the gills of Yesso scallop, glycogenesis, gluconeogenesis and TCA cycle were inhibited, and the glycolysis pathway of glycogen was enhanced to produce more energy for coping with environmental pressure.

Effects of high temperature stress on the intestinal histology and microbiota in Yesso scallop Patinopecten yessoensis

High temperature stress posed by global warming is considered as one of the greatest threats to marine ectotherms by altering their behavior and physiological functions. The intestine and its associated microbiota constitute the first defensive line for the animals against environmental stresses, but their responses to high temperature stress in mollusks are largely unknown. In the present study, the changes of intestinal histology and microbiota were investigated in Yesso scallop Patinopecten yessoensis, a cold-water bivalve species, after high temperature stress. The shrinkage of intestinal lumen, shortening of intestinal villi and increased goblet cells were observed in the intestines of scallops exposed to seawater temperatures of 20 °C (T20 group) and 23 °C (T23 group), compared to the control group (15 °C). High-throughput sequencing of 16S rRNA gene showed that the composition of intestinal microbiota rather than the alpha diversity indices changed significantly after high temperature stress. At the phylum level, the relative abundances of Proteobacteria and Firmicutes decreased progressively with increasing temperature, while that of Bacteroidetes increased by 1.18-fold in the T20 group and 0.95-fold in the T23 group. At the genus level, Tenacibaculum and Mycoplasma were significantly enriched after high temperature stress, and Mycoplasma exhibited highest abundance of 39.43% in the T23 group. Functional prediction revealed that the pathways related to amino acid biosynthesis were blocked after high temperature stress, while that of phospholipases showed the opposite trend. According to the results of network analysis, the network connectivity decreased with increasing temperature, while the percentages of negative correlations in the two high temperature groups were higher than that in the control group. Collectively, the intestinal histology and microbial community of P. yessoensis changed significantly after high temperature stress, which would hinder the nutrient absorption and promote the proliferation of pathogenic microorganisms in the intestine of scallops. Our results will provide novel insights into the occurrence mechanism of mass summer mortality in marine mollusks.

Molecular characterization of myosin heavy chain and its involvement in muscle growth and development in the Yesso scallop Patinopecten yessoensis

Myosin found in scallop adductor muscles is a regulatory protein that is responsible for muscle contraction. Myosin heavy chains (MyHCs) form the core component of myosin and play important roles in myofibril assembly and maintenance. In the present study, we obtained the full-length cDNA sequences encoding MyHC from the Yesso scallop Patinopecten yessoensis and investigated its expression patterns as well as regulatory functioning in muscle growth and development. Py-MyHC was 6054 bp in length and was comprised of a 5′-untranslated region (51 bp), a 3′-untranslated region (180 bp), and an open reading frame (5823 bp) encoding 1940 amino acids. Analysis of the MyHC sequences in bivalves revealed that they were closely homologous to each other, with identities ranging from 74.48% to 95.16%. qPCR analysis and immunohistochemistry staining revealed the existence of tissue-specific expression patterns for Py-MyHC with maximal expression occurring in the striated adductor muscles. Whole-mount in situ hybridization showed that during larval development, Py-MyHC mRNA was mainly expressed in the velum retractor muscles, and in the anterior and posterior adductor muscles. Furthermore, noticeable defects in myofibril structure were found in the striated adductor muscles following the application of RNA interference. Based on the above, it is considered that MyHCs play a crucial role in myofibril assembly and maintenance in both larvae and adults. These findings are considered to provide useful information with regard to the functional basis of muscle growth and muscle production with potential implications for actual aquaculture production.

Effect of water mass mixing on phytoplankton dynamics in the scallop culture areas off Okhotsk Sea: A microcosm experiment

The nearshore (Soya Warm Water (SWW)) and offshore (Intermediate Cold Water (ICW) and Okhostk Sea Low Salinity Water (OSLSW)) waters in the Okhotsk Sea differ substantially in their physico-chemical characteristics. Thus, the extent to which water mass mixing impacts phytoplankton dynamics in the Japanese scallop Patinopecten yessoensis culture areas needs to be assessed to properly manage scallop stocks. This study examined the phytoplankton dynamics in nearshore (SWW) and offshore (ICW and OSLSW) waters and phytoplankton response to water mass mixing (SWW + ICW, OSLSW + ICW) for 5 days in spring (2007) and summer (2007–2009) microcosm experiments. Increased chlorophyll a concentration in ICW was observed due to higher nutrients available for phytoplankton uptake than in OSLSW and SWW at the start of each microcosm experiment. In comparison to spring microcosm, mixing of SWW + ICW promoted higher chlorophyll a concentration and faster Si(OH)4 utilization than in OSLSW + ICW in summer. In addition, the size structure of chlorophyll a differed from micro-size (>10μm) in SWW + ICW to nano-size (<10μm) chlorophyll a in OSLSW + ICW. Suspended particulate organic matter (SPOM) δ13C ratios were positively correlated with chlorophyll a in both mixed microcosms whereas SPOM δ15N isotopic ratios were inversely related to nitrate concentration in SWW + ICW, except in spring. This suggests that stable isotopes can be used in predicting phytoplankton biomass and nutrient utilization. In comparison to microcosm experiments, nearshore water column structure suggests vertical water mixing in summer to autumn when phytoplankton biomass and micro-size fraction were at maximum. Taken together, the advance of nutrient-rich ICW to nutrient-poor nearshore areas in the Okhotsk Sea resulted in increased chlorophyll a biomass and dominance of micro-size phytoplankton in summer, potentially enhancing nutrient and food supply to bottom-cultured scallops as suggested by both microcosm experiments and field observations.

MicroRNA expression signature in the striated and smooth adductor muscles of Yesso scallop Patinopecten yessoensis

Increasing evidences point to the potential role of microRNAs (miRNAs) in muscle growth and development in animals. However, knowledge on the identity of miRNAs and their targets in molluscs remains largely unknown. Scallops have one large adductor muscle, composed of fast (striated) and slow (smooth) muscle types, which display great differences in muscle fibers, meat quality, cell types and molecular components. In the present study, we performed a comprehensive investigation of miRNA transcriptomes in fast and slow adductor muscles of Yesso scallop Patinopecten yessoensis. As a result, 47 differentially expressed miRNAs representing ten miRNA families were identified between the striated and smooth adductor muscles. The KEGG enrichment analysis of their target genes were mainly associated with amino acid metabolism, energy metabolism and glycan biosynthesis. The target genes of miR-133 and miR-71 were validated by the dual-luciferase reporter assays and miRNA antagomir treatment in vivo. The identification and functional validation of these different miRNAs in scallops will greatly help our understanding of miRNA regulatory mechanism that achieves the unique muscle phenotypes in scallops. The present findings provide the direct evidences for muscle-specific miRNAs involved in muscle growth and differentiation in molluscs.

Genome-Wide Identification and Expression Profiling of the COMMD Gene Family in Four Bivalve Molluscs

The COMMD (copper metabolism gene MuRR1 domain) gene family, highly conserved among multicellular eukaryotic organisms, plays important roles in a variety of biological processes, ranging from copper homeostasis, ionic transport, protein trafficking, NF-κB-mediated transcription, and cell proliferation. However, systematic identification, spatiotemporal expression, and stress-responsive patterns of COMMD genes remain obscure in molluscs. Here, we analyzed the characteristics of the COMMD gene family in four bivalve molluscs based on both genome and extensive transcriptomic resources. Firstly, we investigated the genomic signatures, functional domains, and phylogenetic relationships, and ten single-copy members were identified in Yesso scallop (Patinopecten yessoensis), Zhikong scallop (Chlamys farreri), Pacific oyster (Crassostrea gigas), and dwarf surf clam (Mulinia lateralis), respectively. Strong purifying selection was revealed for COMMD4. Higher expressions of most COMMDs were observed in the hepatopancreas, besides which a different tissue preference of COMMDs’ expression was found among four bivalves. Moreover, in the dwarf surf clam, the responses of COMMD members under stresses were found more sensitive in the hepatopancreas than in the gill, and MlCOMMD9 and MlCOMMD4 might be the good candidate stress indicator genes respectively for copper ion stress and V. Anguillarum infection. Our study would contribute to a better understanding for the evolution of the COMMD gene family and provide valuable information for their innate immune roles in bivalve molluscs.

Discovery of Nanos1 and Nanos2/3 as Germ Cell Markers During Scallop Gonadal Development.

Nanos are conserved genes involved in germline cell specification and differentiation. However, little is known about the role of different members of Nanos family in germ cell development in mollusks. In the present study, we conducted genome-wide identification of Nanos family in an economically important scallop Patinopecten yessoensis, and detected their expression in adult tissues and during early development. Two Nanos genes (PyNanos1, PyNanos2/3) were identified, both of which have the N-terminal NOT1-interacting motif and C-terminal (CCHC)2 zinc finger domain. Expression profiles showed that PyNanos1 and PyNanos2/3 were primarily expressed in the gonads, with PyNanos1 being localized in the oogonia, oocytes, and spermatogonia, while PyNanos2/3 being specifically in spermatogonia. The results suggest that PyNanos are germ cell specific and may play crucial roles in gametogenesis in the scallop. PyNanos1 is a maternal gene, which is distributed uniformly at early cleavage, and restricted to 2-3 cell clusters from blastulae to trochophore larvae, suggesting its potential role in the formation of PGCs. Zygotically expressed PyNanos2/3 displayed a similar signal with PyNanos1 in the trochophore larvae, suggesting it may also participate in the formation and/or maintenance of PGCs. This study will benefit germplasm exploitation and conservation in bivalves, and facilitate a better understanding of the evolution of Nanos family and the role of different Nanos in germ cell development in mollusks.

Genetic Evaluation of Growth and Survival-Related Traits in Yesso Scallop Patinopecten yessoensis in Sea-Based Culture System

Worldwide, the bivalve aquaculture industry has realized or recognized the potential gains from selective breeding programs using phenotypic and pedigree data. Yesso scallop Patinopecten yessoensis are among the most important commercial shellfish in China. A family-based breeding program to investigate the genetic variations for growth and survival-related traits at suspended and bottom environments of sea-based culture systems was reported in this study. We proposed and conducted a novel phenotyping technique to longitudinally evaluate the shell heights in the yesso scallop. At harvest after rearing for 20 months, the individual shell heights at 6, 10, 16, and 20 months were simultaneously obtained by the growth rings in the outer shells of the animals. Meanwhile, the body weight (BW) of the survived individuals at harvest was also recorded. Variance components and genetic parameters for growth and survival-related traits were estimated using an animal and threshold model, respectively. In the suspended environment, the heritability estimates for BW and shell heights at specific ages ranged from moderate to high (0.328–0.853). The estimated correlations between shell heights at contiguous ages were consistently high, ranging from 0.890 to 0.958 but decreased with increasing intervals between ages (0.496–0.828). The estimated correlations between shell heights at contiguous ages and BW at harvest were similar, ranging from 0.535 to 0.983. The heritability estimates for individual survival at harvest were at a low level of 0.128 by the probit-threshold model. While at the bottom environment, estimates of heritability for growth and survival-related traits were similar but slightly lower than those at the suspended environment. Furthermore, the genetic correlation for BW between the two environments was very small, which probably indicates genotype-by-environment interaction effects for growth in the yesso scallop. The study can provide prior information, which might develop a new idea for selection in this species. The results are discussed concerning selection work with yesso scallop, and solutions for accurate estimation of genetic parameters and increasing genetic gain are also outlined.