Effects of Vegetation Restoration Age on Soil C:N:P Stoichiometry in Yellow River Delta Coastal Wetland of China

Vegetation restoration can alter carbon(C),nitrogen(N),and phosphorus(P)cycles in coastal wetlands affecting C:N:P stoichiometry.However,the effects of restoration age on soil C:N:P stoichiometry are unclear.In this study,we examined the re-sponses of soil C,N,and P contents and their stoichiometric ratios to vegetation restoration age,focusing on below-ground processes and their relationships to aboveground vegetation community characteristics.We conducted an analysis of temporal gradients based on the'space for time'method to synthesize the effects of restoration age on soil C:N:P stoichiometry in the Yellow River Delta wetland of China.The findings suggest that the combined effects of restoration age and soil depth create complex patterns of shifting soil C:N:P stoichiometry.Specifically,restoration age significantly increased all topsoil C:N:P stoichiometries,except for soil total phosphorus(TP)and the C:N ratio,and slightly affected subsoil C:N:P stoichiometry.The effects of restoration age on the soil C:N ratio was well constrained owing to the coupled relationship between soil organic carbon(SOC)and total nitrogen(TN)contents,while soil TP con-tent was closely related to changes in plant species diversity.Importantly,we found that the topsoil C:N:P stoichiometry was signific-antly affected by plant species diversity,whereas the subsoil C:N:P stoichiometry was more easily regulated by pH and electric con-ductivity(EC).Overall,this study shows that vegetation restoration age elevated SOC and N contents and alleviated N limitation,which is useful for further assessing soil C:N:P stoichiometry in coastal restoration wetlands.

Expression profiles of two small heat shock proteins and antioxidant enzyme activity in Mytilus galloprovincialis exposed to cadmium at environmentally relevant concentrations

Small heat shock proteins encompass a widespread but diverse class of proteins, which play key roles in protecting organisms from various stressors. In the present study, the full-length cDNAs of two small heat shock proteins （MgsHSP22 and MgsHSP24.1） were cloned from Mytilus galloprovincialis, which encoded peptides of 181 and 247 amino acids, respectively. Both MgsHSP22 and MgsHSP24.1 were detected in all tissues examined by real-time PCR, with the highest expression being observed in muscle and gonad tissues. The real-time PCR results revealed that Cd significantly inhibited MgsHSP22 expression at 24 h and MgsHSP24.1 at 24 and 48 h under 5 ug/L Cd2＋ exposure. MgsHSP24.1 expression was also significantly inhibited after 50 ug/L Cd2＋ exposure for 48 h. With regard to antioxidant enzymes, increased GPx and CAT activity were detected under Cd2＋ stress （5 and 50 ug/L）, while no significant difference in SOD activity was observed throughout the experiment. Overall, both MgsHsps and antioxidant enzymes revealed their potential as Cd stress biomarkers in M. galloprovincialis.

Nitrogen Biological Cycle Characteristics of Seepweed(Suaeda salsa) Wetland in Intertidal Zone of Huanghe(Yellow) River Estuary

From April 2008 to November 2009, the nitrogen （N） cycle of plant-soil system in seepweed （Suaeda salsa） wetland in the intertidal zone of the Huanghe （Yellow） River estuary was studied. Results showed that soil N had sig- nificant seasonal fluctuations and vertical distribution, and the net N mineralization rates in topsoil were significantly different in growing season （p 〈 0.01）. The N/P ratio （9.87 ±1.23） of S. salsa was less than 14, indicating that plant growth was limited by N. The N accumulated in S. salsa litter at all times during decomposition, which was ascribed to the N immobilization by microbes from the environment. Soil organic N was the main N stock of plant-soil system, accounting for 97.35% of the total N stock. The N absorption and utilization coefficients of S. salsa were very low （0.0145 and 0.3844, respectively）, while the N cycle coefficient was high （0.7108）. The results of the N turnovers among compartments of S. salsa wetland showed that the N uptake amount of aboveground part and root were 7.764 g/m2and 4.332 g/m2, respectively. The N translocation amounts from aboveground part to root and from root to soil were 3.881 g/m2 and 0.626 g/m2, respectively. The N translocation amount from aboveground living body to litter was 3.883 g/m2, the annual N return amount from litter to soil was more than 0.125（-） g/m2 （minus represented immobili- zation）, and the net N mineralization amount in topsoil （0-15 cm） in growing season was 1.190 g/m2. The assessment of N biological cycle status orS. salsa wetland indicated that N was a very important limiting factor and the ecosystem was situated in unstable and vulnerable status. The S. salsa was seemingly well adapted to the low-nutrient status and vulnerable habitat, and the N quantitative relationships determined in the compartment model might provide scientific base for us to reveal the special adaptive strategy orS. salsa to the vulnerable habitat in the following studies.

Anthropogenic impacts on hyperbenthos in the coastal waters of Sishili Bay,Yellow Sea

A survey on the hyperbenthic communities was carried out in the coastal waters of Sishili Bay in the Yellow Sea in July 2009,to investigate the impact of putative anthropogenic activities related to the presence of a sewage outfall,a harbor and an aquaculture site on the benthic ecosystem.An Agassiz net trawl was used to collect hyperbenthos at 10 sampling stations.Species,diversity,abundance,and biomass were analyzed against water sample data and historical data obtained from records from nearby Yantai City.Fifty-two species were identified in the region,of which Crustacea were the most abundant taxon,followed by Echinodermata.Dominant species included five crustaceans,two echinoderms,one mollusk and one fish species.The results of a BIOENV analysis show that the concentrations of NO3-N and Chl-a were slightly positively correlated with hyperbenthic community structure,while other factors were negatively correlated,including sediment grain size and the percentage of TN and TOC in sediment.Abundance- Biomass Comparison(ABC method) curves indicate that the hyperbenthos in Sishili Bay had been disturbed by putative sources of human activities.Eight out of 10 stations were classified as "moderately perturbed" to "perturbed".

The relationship between soft-bottom macrobenthic communities and environmental variables off Ningjin,eastern Shandong Peninsula

Macrobenthic infauna and associated environmental factors influencing the benthic community in the eastern coastal region of Shandong Peninsula were analyzed in four seasonal surveys from January 2007 to October 2007 （30 stations in winter,20 stations in other three seasons）,in order to understand the community structure and the factors unfluencing the benthic distribution.PRIMER 6.0 and SPSS 15.0 software packages were adopted to analyze the environmental and macrobenthic data.The results show that there were 260 macrobenthic species in total collected from the research region.The composition of species is： Polychaeta （94 species）,Crustacea （75）,Mollusca （56） and Echinoderm （12）,among which,only 23 species were common species in the cruises of every season.The dominant species varied from season to season;however,the polychaete species Paralacydonia paradoxa Fauvel and Echinoderm species Amphioplus japonicus （Matsumoto） were always present year-round.The abundance and biomass of the macrobenthos in the research region were variable from season to season.The results of CLUSTER and MDS analysis show that the similarities of macrobenthic structures between the stations were low;most of the similarities were at about 30% of similarity value,only two stations were up to 70%.In accordance with the similarity values of the macrobenthic structures,we divided the 20 stations into five groups by arbitrary similarity level of 30%.The ABC curve indicates that the marcofauna communities in the research region had not been disturbed massively,except two stations,SB1 and SB3.Ecologically,benthos were controlled by a combination of factors such as salinity,phytoplankton,zooplankton,SiO 3Si and temperature,and no single factor could be considered as an ecological master factor.

Reclamation-oriented spatiotemporal evolution of coastal wetland along Bohai Rim,China

Coastal wetlands are located in the ecotone of interaction between the land surface and sea,and anthropogenic activities extensively interfere with these wetlands through the reclamation of large tidal wetlands and destruction of the function of the ecosystems.In this study,we investigated the dynamic evolutionary characteristics of the Bohai Rim coastal area over the past 40 years using the Modified Normalized Difference Water Index,the fractal dimension,object-oriented classification,the land-use transfer trajectory,and regression analysis.Additionally,we quantified and monitored the evolution of reclamation and analyzed the correlation between reclamation and coastal wetlands based on 99 Landsat-2,-5,and-8 images(at 60 m and 30 m spatial resolution) over the period 1980–2019.The results are as follows.(1) The coastline of the Bohai Rim increased by 1 631.2 km from 1980 to2019 with a zigzag variation.The artificial coastline increased by 2 946.1 km,whereas the natural coastline decreased by 90%.(2) The area of man-made wetlands increased by 3 736.9 km~2,the area of construction land increased by 1 008.4 km~2,and the natural wetland area decreased by 66%.The decrease of tidal flats is the main contributor to the decrease of natural wetland area(takes account for 91.1%).Coastal areas are affected by intense human disturbance,which was taken place across a large area of tidal flats and caused the landscape to fragment and be more heterogeneous.The coastal zone development activities were primarily concentrated in the southern Laizhou Bay,the Yellow River Delta,the Bohai Bay,the northern Liaodong Bay,and the Pulandian Bay.The solidified shorelines and increase in sea level have resulted in intertidal wetlands decreasing and impaired wetland ecology.(3) There is a good agreement between reclamation and the size of the coastal wetlands.Both land reclamation and the reduction in coastal wetland areas are significantly related to the population size,fishery output value,and urbanization rate.In summary,human activities,such as the construction of aquaculture ponds and salt pans,industrialization,and urbanization,are the primary forces that influence the environmental changes in the coastal region.This study is beneficial for establishing and improving the systems for the rational development and utilization of natural resources,and provides theoretical references for restoring wetland ecology and managing future reclamation activities in other coastal zone-related areas.

Climatology of Winter Extratropical Cyclones over the Coastal Waters of China

The East China coast is one major cyclogenesis region of extratropical cyclones (ETCs) in boreal winter. In this study, the climatological characteristics of winter ETCs passing over the coastal water of China are analyzed by using clustering and composite analysis. Automated cyclone detection and tracking algorithm are used to identify the ETCs, which are further classified into subgroups according to their trajectory features by using the probabilistic clustering algorithm. Six distinct types of straight moving or recurving trajectories with different climatological characteristics are identified in the clustering analysis. Accordingly, the composite analyses also present six distinct synoptic patterns corresponding to cyclogenesis as well as the impact of ETCs on terrestrial precipitation. The spatio-temporal characteristics of winter ETCs’ origination, movement, and impact are revealed by the clustering and composite analyses.

Temporal and Spatial Distribution of DIN and DIP Concentrations and Source Apportionment Along the Bohai Sea of China During 2015-2022

The Bohai Sea is one of the most polluted sea areas in China.In this study,we used 2184 integrated concentrations of dissolved inorganic nitrogen(DIN)and dissolved inorganic phosphorus(DIP)in the Bohai Sea of China during spring(March,April,and May),summer(June,July,and August),and autumn(October and November)from 2015 to 2022 to explore the trends and sources of nutrients variations.From 2015 to 2022,DIN showed a downward trend until 2020 and then an upward trend,whereas DIP exhibited a stable trend with a slight decrease.The concentrations of DIN and DIP had similar seasonal pattern which was the highest in autumn(0.292±0.247 mg/L for DIN and 0.013±0.016 mg/L for DIP)but lower in spring(0.267±0.238 mg/L for DIN and 0.006±0.010 mg/L for DIP)and summer(0.263±0.324 mg/L for DIN and 0.008±0.010 mg/L for DIP).Sources of DIN and DIP apportioned by the positive matrix factorization(PMF)model were riverine input,sediment resuspension,sewage discharge,atmospheric deposition,and underground input.During 2015-2022,the largest contributor to DIN was sewage discharge(28.7%)and the largest contributor to DIP was sediment resuspension(44.6%).Seasonally,DIN in spring and autumn was dominated by sewage discharge(45.4%and 27.8%,re-spectively).Whereas in summer,it was dominated by riverine input(32.4%)and atmospheric deposition(29.7%).DIP was dominated by sediment resuspension during all three seasons(35.8%-52.5%).In addition,the increase in DIN concentrations in 2021 and 2022 were mainly due to the incremental input of river discharge and atmospheric deposition caused by increased precipitation during sum-mer and autumn.

Application of Stable Isotope Techniques in Studies of Carbon and Nitrogen Biogeochemical Cycles of Ecosystem

Stable isotope techniques have been proved useful as tools for studying the carbon (C) and nitrogen (N) biogeochemical cycles of ecosystem. This paper firstly introduced the basic principles and the distribution characteristics of stable isotope, then reviewed the recent advances and applications of stable isotope in the C and N biogeochemical cycles of ecosystem. By applying the 13 C natural abundance technique, ecologists are able to understand the photosynthetic path and CO 2 fixation of plants, the CO 2 exchange and C balance status of ecosystem, the composition, distribution and turnover of soil organic C and the sources of organic matter in food webs, while by using the 13 C labeled technique, the effects of elevated CO 2 on the C processes of ecosystem and the sources and fate of organic matter in ecosystem can be revealed in detail. Differently, by applying the 15 N natural abundance technique, ecologists are able to analyze the biological N 2 -fixation, the N sources of ecosystem, the N transformation processes of ecosystem and the N trophic status in food webs, while by using the 15 N labeled technique, the sources, transformation and fate of N in ecosystem and the effects of N input on the ecosystem can be investigated in depth. The applications of both C and N isotope natural abundance and labeled techniques, combined with the elemental, other isotope ( 34 S) and molecular biomarker information, will be more propitious to the investigation of C and N cycle mechanisms. Finally, this paper concluded the problems existed in current researches, and put forward the perspective of stable isotope techniques in the studies on C and N biogeochemical cycles of ecosystem in the future.

The influence of salinity on toxicological effects of arsenic in digestive gland of clam Ruditapes philippinarum using metabolomics

Ruditapes philippinarum,a clam that thrives in intertidal zones of various salinities,is a useful biomonitor to marine contaminants.We investigated the influence of dilution to 75% and 50% of normal seawater salinity(31.1) on the responses of the digestive gland of R.philippinarum to arsenic exposure(20 μg/L),using nuclear magnetic resonance(NMR)-based metabolomics.After acute arsenic exposure for 48 h,salinity-dependent differential metabolic responses were detected.In normal seawater,arsenic exposure increased the concentrations of branched-chain amino acids,and of threonine,proline,phosphocholine and adenosine,and it decreased the levels of alanine,hypotaurine,glucose,glycogen and ATP in the digestive glands.Differential changes in metabolic biomarkers observed at lower salinity(～23.3) included elevation of succinate,taurine and ATP,and depletion of branched-chain amino acids,threonine and glutamine.Unique effects of arsenic at the lowest salinity(～15.6) included down-regulation of glutamate,succinate and ADP,and up-regulation of phosphocholine.We conclude that salinity influences the metabolic responses of this clam to arsenic.

Temporal and spatial variations in the distribution of macroalgal communities along the Yantai coast, China

To explore the impact of environmental variables on macroalgal communities, their temporal and spatial distributions were examined along the Yantai coast, China between April 2010 and March 2011. Macroalgae sampling was conducted monthly at four sites along the coast: Jiahe River estuary, Zhifu Island, Fisherman's Wharf, and Yangma Island. The species composition and abundance, and their relationship with environmental variables were assessed. Along the Yantai coast, 35 macroalgae species were identifi ed, including 24 Rhodophyta spp., 7 Chlorophyta, and 4 Phaeophyta spp. Highest species numbers were recorded in the summer at all sampling sites, except in the Jiahe River estuary. Macroalgae biomass was the greatest in the summer. Year-round, the highest species number and dry biomass recorded at Fisherman's Wharf and Yangma Island was attributed to the substrate type. In summer, Ulva pertusa Kjellman was the dominant species identifi ed along the Yantai coast, which indicates a risk of macroalgae blooms. Our results show that seawater temperature and nutrients appear to signifi cantly affect the temporal and spatial patterns of macroalgal abundance along the Yantai coast. The effects of environmental variables on the macroalgae on the Yantai coast need further study.

Distribution of diatoms and silicoflagellates in surface sediments of the Yellow Sea and offshore from the Changjiang River, China

The spatial distribution of siliceous microfossils （diatoms and silicoflagellates） in the surface sediments was mapped at 113 sites in the Yellow Sea and sea areas adjacent to the Chang^iang （Yangtze） River, China. In total, 267 diatom taxa and two silicoflagellate species were identified from the sediments. The spatial variations in abundance and diversity were classified into three distinct geographic patterns using Q mode clustering： a south-north geographic pattern, a coastal-offshore pattern and a unique pattern in the Changjiang River mouth. The south-north geographic pattern was related to the spatial variations in sea temperature. Coscinodiscus oculatus, a warm-water species, indicated these variations by a gradual decrease in abundance from the south to the north. The coastal-offshore pattern was in response to the spatiaJ variations in salinity. Cyclotella stylorum, Actinocyclus ehrenbergii and Dictyocha messanensis, the dominant brackish species in coastal waters, significantly decreased at the isobaths of approximately 30 m, where the salinity was higher than 31. Paralia sulcata and Podosira stelliger indicated the impact of the Yellow Sea Warm Current in the central Yellow Sea. The unique pattern in the Changjiang River mouth showed the highest species diversity but lower abundance, apparently because： freshwater input can significantly increase the proportion of brackish species; nutrients can supply the growth ofphytoplankton; and high sedimentation rates can dilute the mierofossil abundance in the sediments. Our results show that an integration of environmental factors （e.g., nutrient levels, sedimentation rate, sea temperature, salinity and water depth） determined the spatial characteristics of the siliceous micro fossils in the surface sediments.

Glutathione S-transferase(GST) gene expression profiles in two marine bivalves exposed to BDE-47 and their potential molecular mechanisms

Glutathione S-transferases （GSTs） are phase II enzymes that facilitate the detoxification of xenobioties and play important roles in antioxidant defense. We investigated the expression patterns of seven Venerupis philippinarum GSTs （VpGSTs） and four Mytilus galloprovincialis GSTs （MgGSTs） following exposure to BDE-47. Differential expressions of the seven VpGSTs and four MgGSTs transcripts were observed, with differences between the hepatopancreas and gills. Among these GSTs, the sigma classes （VpGSTS1, VpGSTS2, VpGSTS3, MgGST1, and MgGST3） were highly expressed in response to BDE-47 exposure, demonstrating their potential as molecular biomarkers for environmental biomonitoring studies. We obtained the three-dimensional crystal structures of VpGSTs and MgGSTs by homologous modeling. A model to elucidate the binding interactions between the ligands and receptors was defined by molecular docking, Hydrophobic and n were the most often observed interactions between BDE-47 and the GSTs.

Effect of cadmium on the defense response of Pacific oyster Crassostrea gigas to Listonella anguillarum challenge

Heavy metal pollution can affect the immune capability of organisms. We evaluated the effect of cadmium （Cd） on the defense responses of the Pacific oyster Crassostrea gigas to Listonella anguillarum challenge. The activities of several important defensive enzymes, including superoxide dismutase （SOD）, glutathione peroxidase （GPx）, acid phosphatase （ACP）, Na＋, K＋-ATPase in gills and hepatopancreas, and phenoloxidase-like （POL） enzyme in hemolymph were assayed. In addition, the expression levels of several genes, including heat shock protein 90 （IrtSP9~））, metallothionein （MT）, and bactericidal/permeability increasing （BPI） protein were quantified by fluorescent quantitative PCR. The enzyme activities of SOD, ACP, POL, and GPx in hepatopancreas, and the expression of HSP90 were down-regulated, whereas GPx activity in the gill, Na＋, K＋-ATPase activities in both tissues, and MT expression was increased in Cd- exposed oysters post L. anguillarum challenge. However, BPI expression was not significantly altered by co-stress of L. anguillarum infection and cadmium exposure. Our results suggest that cadmium exposure alters the oysters＇ immune responses and energy metabolism following vibrio infection.

Runoff variation affected by precipitation change and human activity in Hailiutu River basin,China

For recent years,runoff generation and hydrological processes in Hailiutu River basin have been greatly changed by climate change and human activity,especially water and soil conservation construction.In this study,the trends in precipitation,evapotranspiration(ET)and river runoff as well as the effects of precipitation change and human activity on runoff variation have been studied.The results showed that during 1960-2000,annual precipitation and river runoff,monthly precipitation and ET in September and October as well as monthly runoff in all months showed a significant decrease.In addition,peak flow and base flow had a large decrease.Under the joint influence of precipitation change and human activity,the mean annual runoff decreased by 35 million m3 from the baseline period(1960-1985)to the change period(1986-2000),which accounted for 60.9%and 39.1%of the total runoff decrease,respectively.Precipitation change played a primary role in the decrease of annual runoff whereas human activity,particularly water and soil conservation construction,also had remarkable impacts on runoff variation.

Homologous cloning, characterization and expression of a new halophyte phytochelatin synthase gene in Suaeda salsa

The halophyte Suaeda salsa can grow in heavy metal-polluted areas along intertidal zones having high salinity.Since phytochelatins can effectively chelate heavy metals,it was hypothesized that S.salsa possessed a phytochelatin synthase(PCS) gene.In the present study,the cDNA of PCS was obtained from S.salsa(designated as SsPCS) using homologous cloning and the rapid amplification of cDNA ends(RACE).A sequence analysis revealed that SsPCS consisted of 1 916 bp nucleotides,encoding a polypeptide of 492 amino acids with one phytochelatin domain and one phytochelatin C domain.A similarity analysis suggested that SsPCS shared up to a 58.6%identity with other PCS proteins and clustered with PCS proteins from eudicots.There was a new kind of metal ion sensor motif in its C-terminal domain.The SsPCS transcript was more highly expressed in elongated and fibered roots and stems(P<0.05) than in leaves.Lead and mercury exposure significantly enhanced the mRNA expression of SsPCS(P<0.05).To the best of our knowledge,SsPCS is the second PCS gene cloned from a halophyte,and it might contain a different metal sensing capability than the first PCS from Thellungiella halophila.This study provided a new view of halophyte PCS genes in heavy metal tolerance.

Molecular cloning and differential expression patterns of sigma and omega glutathione S-transferases from Venerupis philippinarum to heavy metals and benzo[a]pyrene exposure

Glutathione S-transferases (GSTs) are a class of enzymes that facilitate the detoxification of xenobiotics, and also play important roles in antioxidant defense. We identified two glutathione S-transferase isoforms (VpGSTS, sigma GST; VpGSTO, omega GST) from Venerupis philippinarum by RACE approaches. The open reading frames of VpGSTS and VpGSTO were of 612 bp and 729 bp, encoding 203 and 242 amino acids with an estimated molecular mass of 22.88 and 27.94 kDa, respectively. The expression profiles of VpGSTS and VpGSTO responded to heavy metals and benzo[a]pyrene (B[a]P) exposure were investigated by quantitative real-time RT-PCR. The expression of VpGSTS and VpGSTO were both rapidly up-regulated, however, they showed differential expression patterns to different toxicants. Cd displayed stronger induction of VpGSTS expression with an approximately 12-fold increase than that of VpGSTO with a maximum 6.4-fold rise. Cu exposure resulted in similar expression patterns for both VpGSTS and VpGSTO. For B[a]P exposure, the maximum induction of VpGSTO was approximately two times higher than that of VpGSTS. Altogether, these findings implied the involvement of VpGSTS and VpGSTO in host antioxidant responses, and highlighted their potential as a biomarker to Cd and B[a]P exposure.

Source diagnostic and weathering indicators of oil spills utilizing bicyclic sesquiterpanes

Bicyclic Sesquiterpanes （BSs） are ubiquitous components of crude oils and ancient sediments. BSs in crude oils and diesel oil were identified and characterized, the effects of moderate weathering on BS distributions were discussed, and a methodology using diagnostic indices of BSs was developed for oil correlation and dif- ferentiation. The results showed that crude oils and oil products had different abundances and distributions of BSs and, consequently, resulted in different diagnostic ratios. The selected diagnostic ratio indices of BSs, such as BS4/BS5 （ratio of 4,4,8,9,9-pentamethyl-decahydronaphthalene to 8/3 （H）-drimane）, BS6/BS5 （ratio of 4,4,9,9,10-pentamethyl-decahydronaphthalene to 8/3 （H）-drimane）, BS8/BS9 （ratio of the second peak to the third peak of C16 sequiterpane）, and BS8/BSIO （ratio of C16 sequiterpane to 8fl （H）-homodrimane）, still maintained better stabilities （%RSD 〈 5%） after weathering for 30 d. The longer weathering process （150 d） had some effect on such ratios （5% 〈 %RSD 〈 10%）. The facts of the uniqueness, abundance in petroleum, and chemical stability of BSs enable them to be suitable as an effective diagnostic means for identifying spilled oil with moderate weathering, particularly for lighter refined product samples that are difficult to identify by current techniques.

Molecular cloning, characterization and expression profiles of thioredoxin 1 and thioredoxin 2 genes in Mytilus galloprovincialis

Thioredoxin （Trx） proteins are involved in many biological processes especially the regulation of cellular redox homeostasis. In this study, two Trx cDNAs were cloned from the mussel Mytilus galloprovincialis using rapid amplification of cDNA ends-polymerase chain reaction （RACE-PCR）. The two cDNAs were named MgTrxl and MgTrx2, respectively. The open reading frames of MgTrxl and MgTrx2 were 318 and 507 base pairs （bp） and they encoded proteins of 105 and 168 amino acids with estimated molecular masses of 11.45 and 18.93 kDa, respectively. Sequence analysis revealed that both proteins possessed the conserved active site dithiol motif Cys-Gly-Pro-Cys. In addition, MgTrx2 also possessed a putative mitochondrial targeting signal suggesting that it is located in the mitochondria. Quantitative real-time polymerase chain reaction （qPCR） revealed that both MgTrxl and MgTrx2 were constitutively expressed in all tissues examined. The MgTrxl transcript was most abundant in hemocytes and gills, whereas the MgTrx2 transcript was most abundant in gonad, hepatopancreas, gill and hemocytes. Following Vibrio anguillarum challenge, the expression of MgTrxl was up-regulated and reached its peak, at a value 10-fold the initial value, at 24 h. Subsequently, expression returned back to the original level. In contrast, the expression level of MgTrx2 was down-regulated following bacterial stimulation, with one fifth of the control level evident at 12 h post challenge. These results suggest that MgTrxl and MgTrx2 may play important roles in the response ofM. galloprovincialis to bacterial challenge.

Analyzing biases of nitrogen contents and δ^(15)N values arising from acidified marine sediments with different CaCO_3 concentrations

In some studies, the researchers pretreated and measured organic carbon, nitrogen and their isotopes（δ^（13）C and δ^（15）N） of marine sediment together, to save costs and resources of analysis. However, the procedure of acidification to remove inorganic carbon for analysing δ^（13）C can affect the values of nitrogen and δ^（15）N, and the biases vary a lot depending on the CaCO_3 contents of sediments. In this study, the biases of total nitrogen（TN）and δ^（15）N values arising from acidified sediments were compared between the CaCO_3-poor（1%–16%） and CaCO_3-rich（20%–40%） samples. TN and δ^（15）N values were altered during acid treatment（without centrifugation） that possibly led to N-containing compounds volatilization. For CaCO_3-poor samples, acidification led to a range of 0%–40% TN losses and 0‰–2‰ shift in δ^（15）N values; and 10%–60% TN losses and 1‰–14‰ shift in δ^（15）N values for CaCO_3-rich samples. The biases from most samples exceeded the precision of the instrument（0.002% for TN and 0.08‰ for δ^（15）N）, and high biases could mislead our judgment for the environmental implication of the data. Thus,avoiding co-analysis of organic carbon, nitrogen and their isotopes（δ^（13）C and δ^（15）N） in sediments, even for CaCO_3-poor marine sediments, is necessary.