Posts Tagged 'molecular biology'

Ocean acidification alters the burrowing behaviour, Ca2+/Mg2+-ATPase activity, metabolism, and gene expression of a bivalve species, Sinonovacula constricta

Although the impacts of ocean acidification on fertilization, embryonic development, calcification, immune response, and behaviour have been well studied in a variety of marine organisms, the physiological and molecular mechanism manifesting acidification stress on behavioural response remains poorly understood. Therefore, the impacts of future ocean acidification scenarios (pH at 7.8, 7.6, and 7.4) on the burrowing behaviour, Ca2+/Mg2+-ATPase activity, metabolism, and expression of energy-producing-related genes of the razor clam Sinonovacula constricta were investigated in the present study. The results showed that elevated CO2 partial pressure ( pCO2) (pH at 7.6 and 7.4) led to a significant reduction in the digging depth of the razor clam. In addition, exposure to pCO2-acidified seawater depressed the metabolism and activity of Ca2+/Mg2+-ATPase, which may partially contribute to the reduced digging depth detected. Furthermore, the expression of energy-producing-related genes was generally induced by exposure to acidified seawater and could be accounted for by an increased energy demand under acidification stress. The results obtained suggest ocean acidification may exert a behavioural impact through altering physiological condition in the razor clam.

Continue reading ‘Ocean acidification alters the burrowing behaviour, Ca2+/Mg2+-ATPase activity, metabolism, and gene expression of a bivalve species, Sinonovacula constricta’

Genome-wide identification, characterization and expression analyses of TLRs in Yesso scallop (Patinopecten yessoensis) provide insight into the disparity of responses to acidifying exposure in bivalves

  Highlights

  • Eighteen TLR superfamily members were identified in the P. yessoensis genome.
  • Phylogenetic analysis confirmed duplication and expansion of TLR genes in mollusk.
  • The 18 PyTLRs showed different immune response patterns to acidifying exposure.
  • Adaptive recruitment of tandem duplication of TLR genes have been arisen to the immune stress.

Abstract

Toll-like receptors (TLRs) play a crucial role in innate immunity by recognizing specific pathogen-associated molecular patterns, including lipoproteins, lipopeptides, lipopolysaccharide, flagellin, dsRNA, ssRNA and CpG DNA motifs. Although significant effects of TLRs on immunity have been reported in most vertebrates and some invertebrates, the complete TLR superfamily has not been systematically characterized in scallops. In this study, 18 TLR genes were identified from Yesso scallop (Patinopecten yessoensis) using whole-genome scanning. Phylogenetic and protein structural analyses were performed to determine the identities and evolutionary relationships of the 18 genes. Extensive expansion of TLR genes from the Yesso scallop genome indicated gene duplication events. In addition, expression profiling of PyTLRs was performed at different acidifying exposure levels (pH = 6.50, 7.50) with different challenge durations (3, 6, 12 and 24 h) via in silico analysis using transcriptome and genome databases. Our results confirmed the inducible expression patterns of PyTLRs under acidifying exposure, and the responses to immune stress may have arisen through adaptive recruitment of tandem duplications of TLR genes. Collectively, this study provides novel insight into PyTLRs as well as the specific role and response of TLR signaling pathways in host immune responses against acidifying exposure in bivalves.

 

Continue reading ‘Genome-wide identification, characterization and expression analyses of TLRs in Yesso scallop (Patinopecten yessoensis) provide insight into the disparity of responses to acidifying exposure in bivalves’

The development of contemporary European sea bass larvae (Dicentrarchus labrax) is not affected by projected ocean acidification scenarios

Ocean acidification is a recognized consequence of anthropogenic carbon dioxide (CO2) emission in the atmosphere. Despite its threat to marine ecosystems, little is presently known about the capacity for fish to respond efficiently to this acidification. In adult fish, acid–base regulatory capacities are believed to be relatively competent to respond to hypercapnic conditions. However, fish in early life stage could be particularly sensitive to environmental factors as organs and important physiological functions become progressively operational during this period. In this study, the response of European sea bass (Dicentrarchus labrax) larvae reared under three ocean acidification scenarios, i.e., control (present condition, P CO 2   PCO2  = 590 µatm, pH total = 7.9), low acidification (intermediate IPCC scenario, P CO 2   PCO2  = 980 µatm, pH total = 7.7), and high acidification (most severe IPCC scenario, P CO 2   PCO2  = 1520 µatm, pH total = 7.5) were compared across multiple levels of biological organizations. From 2 to 45 days-post-hatching, the chronic exposure to the different scenarios had limited influence on the survival and growth of the larvae (in the low acidification condition only) and had no apparent effect on the digestive developmental processes. The high acidification condition induced both faster mineralization and reduction in skeletal deformities. Global (microarray) and targeted (qPCR) analysis of transcript levels in whole larvae did not reveal any significant changes in gene expression across tested acidification conditions. Overall, this study suggests that contemporary sea bass larvae are already capable of coping with projected acidification conditions without having to mobilize specific defense mechanisms.

Continue reading ‘The development of contemporary European sea bass larvae (Dicentrarchus labrax) is not affected by projected ocean acidification scenarios’

Genome-wide transcriptional reprogramming in the seagrass Cymodocea nodosa under experimental ocean acidification

Here we report the first use of massive scale RNA-Sequencing to explore seagrass response to CO2-driven ocean acidification (OA). Large-scale gene expression changes in the seagrass Cymodocea nodosa occurred at CO2 levels projected by the end of the century. C. nodosatranscriptome was obtained using Illumina RNA-Seq technology and de novo assembly, and differential gene expression was explored in plants exposed to short-term high CO2 / low pH conditions. At high pCO2, there was a significant increased expression of transcripts associated to photosynthesis, including light reaction functions and CO2 fixation, and also to respiratory pathways, specifically for enzymes involved in glycolysis, in the tricarboxylic acid cycle and in the energy metabolism of the mitochondrial electron transport. The up-regulation of respiratory metabolism is probably supported by the increased availability of photosynthates and increased energy demand for biosynthesis and stress-related processes under elevated CO2 and low pH. The up-regulation of several chaperones resembling heat stress-induced changes in gene expression, highlighted the positive role these proteins play in tolerance to intracellular acid stress in seagrasses. OA further modifies C. nodosa secondary metabolism inducing the transcription of enzymes related to carbon-based-secondary compounds biosynthesis, in particular the synthesis of polyphenols and isoprenoid compounds that have a variety of biological functions including plant defense. By demonstrating which physiological processes are most sensitive to OA, this research provide a major advance in the understanding of seagrass metabolism in the context of altered seawater chemistry from global climate change.

Continue reading ‘Genome-wide transcriptional reprogramming in the seagrass Cymodocea nodosa under experimental ocean acidification’

Impacts of a reduction in seawater pH mimicking ocean acidification on the structure and diversity of mycoplankton communities

Increases in atmospheric carbon dioxide (CO2) change ocean chemistry, as dissolved CO2 leads to a reduction in the seawater pH. Many marine taxa have been shown to be affected by ocean acidification; however, information on marine fungi is lacking. We analyzed the effect of pH on mycoplankton communities. The pH of microcosms was adjusted to a value mimicking the predicted ocean acidification in the near future. Fungal communities were analyzed using a double-marker gene approach, allowing a more detailed analysis of their response using 454 pyrosequencing. Mycoplankton communities in microcosms with in situ and adjusted water pH values differed significantly in terms of structure and diversity. The differences were mainly abundance shifts among the dominant taxa, rather than the exclusion of fungal groups. A sensitivity to lower pH values was reported for several groups across the fungal kingdom and was not phylogenetically conserved. Some of the fungal species that dominated the communities of microcosms with a lower pH were known pathogenic fungi. With the increasing awareness of the significant role fungi play in marine systems, including performing a diverse range of symbiotic activities, our results highlight the importance of including fungi in further research projects studying and modeling biotic responses to the predicted ocean acidification.

Continue reading ‘Impacts of a reduction in seawater pH mimicking ocean acidification on the structure and diversity of mycoplankton communities’

A shell-formation related carbonic anhydrase in Crassostrea gigas modulates intracellular calcium against CO2 exposure: implication for impacts of ocean acidification on mollusk calcification

Ocean acidification (OA) could decrease the shells and skeletons formation of mollusk by reducing the availability of carbonate ions at calcification sites. Carbonic anhydrases (CAs) convert CO2 to HCO3− and play important roles in biomineralization process from invertebrate to vertebrate. In the present study, a CA (designated as CgCA) was identified and characterized in Pacific oyster C. gigas. The cDNA of CgCA was of 927 bp encoding a predicted polypeptide of 308 amino acids with a signal peptide and a CA catalytic function domain. The mRNA transcripts of CgCA were constitutively expressed in all tested tissues with the highest levels in mantle and hemocytes. During the early development period, the mRNA transcripts of CgCA could be detected in all the stages with the highest level in D-veliger larvae. Elevated CO2 increased the mRNA transcripts of CgCA in muscle, mantle, hepatopancreas, gill and hemocytes significantly (p < 0.05) and induced the translocation of CgCA in hemocytes and mantle. Moreover, elevated CO2 also caused the decrease of intracellular Ca2+ in hemocytes (p < 0.05). The inhibition of CA by acetazolamide and suppression of CgCA gene via RNA interference could increase the intracellular Ca2+ in hemocytes (p < 0.05). Besides, the decrease of intracellular Ca2+ content caused by Ca2+ reagent ionomycin could affect localization of CgCA in mantle tissue. The results indicated CgCA played essential roles in calcification and elevated CO2 accelerated the mutual modulation between calcium and CgCA, implying reduced calcification rate and dissolved shells under OA.

Continue reading ‘A shell-formation related carbonic anhydrase in Crassostrea gigas modulates intracellular calcium against CO2 exposure: implication for impacts of ocean acidification on mollusk calcification’

Influencing mechanism of ocean acidification on byssus performance in the pearl oyster Pinctada fucata

The byssus is an important adhesive structure by which bivalves robustly adhere to underwater substrates. It is susceptible to carbon dioxide-driven ocean acidification (OA). Previous investigations have documented significant adverse effects of OA on the performance of byssal threads, but the mechanisms remain largely unknown. In this study, multiple approaches were employed to reveal the underlying mechanisms for the effects of OA on byssus production and mechanical properties in the pearl oyster Pinctada fucata. The results showed that OA altered the abundance and secondary structure of byssal proteins and affected the contents of metal ions in distal threads, which together reduced the byssus diameter and amplified byssus nanocavity, causing reductions in mechanical properties (strength and extensibility). Expression analysis of key foot protein genes further confirmed changes in byssal protein abundance. Moreover, comparative transcriptome analysis revealed enrichment of ion transportation- and apoptosis-related categories, up-regulation of apoptosis-related pathways and down-regulation of the “extracellular matrix-receptor interaction” pathway, which may influence foot locomotion physiology, leading to a decrease in byssus production. This study provides mechanistic insight into the effects of OA on pearl oyster byssus, which should broaden our overall understanding of the impacts of OA on marine ecosystem.

Continue reading ‘Influencing mechanism of ocean acidification on byssus performance in the pearl oyster Pinctada fucata’


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