Complementary DNA sequencing (cDNA): an eff ective approach for assessing the diversity and distribution of marine benthic ciliates along hydrographic gradients

The Yellow Sea Cold Water Mass(YSCWM)is a distinct hydrographic phenomenon of the Yellow Sea,and the distribution pattern of meio-and macrobenthos diff ers inside and outside of the YSCWM.However,such a pattern has never been observed in the microbenthic ciliate communities.Therefore,we hypothesized that benthic ciliates followed a similar distribution pattern as meio-and macrobenthos,but this pattern has not been uncovered by morphological methods.We evaluated the diversity and distribution of benthic ciliates at fi ve stations along hydrographic gradients across the YSCWM and adjacent shallow water by using morphology and DNA and complementary DNA(cDNA)high-throughput sequencing of the V4 region of 18S rRNA gene.Results showed that the diversity of benthic ciliates detected by DNA(303 OTUs),and the cDNA(611 OTUs)sequencing was much higher than that detected by the morphological method(79 species).Morphological method detected roughly diff erent ciliate communities inside and outside of the YSCWM,but without statistical signifi cance.No clear pattern was obtained by DNA sequencing.In contrast,cDNA sequencing revealed a distinct distribution pattern of benthic ciliate communities like meioand macrobenthos,which coincided well with the results of the environmental parameter analysis.More than half of the total sequences detected by DNA sequencing belonged to planktonic ciliates,most(if not all)of which were recovered from historic DNA originating through the sedimentation of pelagic forms because none of them were observed morphologically.The irrelevant historic DNA greatly infl uenced the recovery of rare species and thus limited the understanding of the benthic ciliate diversity and distribution.Our research indicates that the methods used have signifi cant eff ects on the investigation of benthic ciliate communities and highlights that cDNA sequencing has great advantages in estimating the diversity and distribution of benthic ciliates,as well as the potential for benthic environmental assessments.

Full T-cell activation and function in teleosts require collaboration of first and co-stimulatory signals

Mammalian T-cell responses require synergism between the first signal and co-stimulatory signal.However,whether and how dual signaling regulates the T-cell response in early vertebrates remains unknown.In the present study,we discovered that the Nile tilapia(Oreochromis niloticus)encodes key components of the LAT signalosome,namely,LAT,ITK,GRB2,VAV1,SLP-76,GADS,and PLC-γ1.These components are evolutionarily conserved,and CD3εmAb-induced T-cell activation markedly increased their expression.Additionally,at least ITK,GRB2,and VAV1 were found to interact with LAT for signalosome formation.Downstream of the first signal,the NF-κB,MAPK/ERK,and PI3K-AKT pathways were activated upon CD3εmAb stimulation.Furthermore,treatment of lymphocytes with CD28 mAbs triggered the AKT-mTORC1 pathway downstream of the co-stimulatory signal.Combined CD3εand CD28 mAb stimulation enhanced ERK1/2 and S6 phosphorylation and elevated NFAT1,c-Fos,IL-2,CD122,and CD44 expression,thereby signifying T-cell activation.Moreover,rather than relying on the first or co-stimulatory signal alone,both signals were required for T-cell proliferation.Full T-cell activation was accompanied by marked apoptosis and cytotoxic responses.These findings suggest that tilapia relies on dual signaling to maintain an optimal T-cell response,providing a novel perspective for understanding the evolution of the adaptive immune system.

Anti-infective immune functions of type Ⅳ interferon in grass carp(Ctenopharyngodon idella):A novel antibacterial and antiviral interferon in lower vertebrates

Type Ⅳ interferon(IFN-υ)is a recently discovered cytokine crucial for host defense against viral infections.However,the role and mechanisms of IFN-υin bacterial infections remain unexplored.This study investigated the antibacterial and antiviral functions and mechanisms of grass carp(Ctenopharyngodon idella)IFN-υ(CiIFN-υ)both in vivo and in vitro.The CiIFN-υgene was first identified and characterized in grass carp.Subsequently,the immune expression of CiIFN-υsignificantly increased following bacterial challenge,indicating its response to bacterial infections.The eukaryotic recombinant expression plasmid of CiIFN-υwas then constructed and transfected into fathead minnow(FHM)cells.Supernatants were collected and incubated with four bacterial strains,followed by plate spreading and colony counting.Results indicated that CiIFN-υexhibited more potent antibacterial activity against gram-negative bacteria compared to gram-positive bacteria and aggregated gram-negative bacteria but not gram-positive bacteria.In vivo experiments further confirmed the antibacterial function,showing high survival rates,low tissue edema and damage,reduced tissue bacterial load,and elevated proinflammatory response at the early stages of bacterial infection.In addition,the antiviral function of CiIFN-υwas confirmed through in vitro and in vivo experiments,including crystal violet staining,survival rates,tissue viral burden,and reverse transcription-quantitative real-time polymerase chain reaction(RT-qPCR).This study highlights the antibacterial function and preliminary mechanism of IFN-υ,demonstrating that IFN-υpossesses dual functions against bacterial and viral infections.

Combining genome-wide association study and transcriptome analysis to identify molecular markers and genetic basis of population-asynchronous ovarian development in Coilia nasus

Coilia nasus,a migratory fish species found in the middle and lower reaches of the Yangtze River and along offshore areas of China,possesses considerable aquacultural and economic potential.However,the species faces challenges due to significant variation in the gonadal development rate among females,resulting in inconsistent ovarian maturation times at the population level,an extended reproductive period,and limitations on fish growth rate due to ovarian prematurity.In the present study,we combined genome-wide association study(GWAS)and comparative transcriptome analysis to investigate the potential single nucleotide polymorphisms(SNPs)and candidate genes associated with population-asynchronous ovarian development in C.nasus.Genotyping of the female population based on whole-genome resequencing yielded 2120695 high-quality SNPs,39 of which were suggestively associated with ovarian development.Of note,a significant SNP peak on LG21 containing 30 suggestively associated SNPs was identified,with cpne5a determined as the causal gene of the peak.Therefore,single-marker and haplotype association analyses were performed on cpne5a,revealing four genetic markers(P<0.05)and seven haplotypes(r2>0.9)significantly associated with the phenotype.Comparative transcriptome analysis of precociously and normally maturing individuals screened out 29 and 426 overlapping differentially expressed genes in the brain and ovary,respectively,between individuals of different body sizes.Integrating the GWAS and transcriptome analysis results,this study identified genes and pathways related to hypothalamic-pituitary-gonadal axis hormone secretion,extracellular matrix,angiogenesis,and gap junctions involved in population-asynchronous ovarian development.The insights gained from this study provide a basis for a deeper understanding of the molecular mechanisms underlying ovarian development in fish and may facilitate the genetic breeding of C.nasus strains exhibiting population-synchronous ovarian development in the future.

Genetic analysis of selected Sargassum fusiforme (Harvey) Setchell (Sargassaceae, Phaeophyta) strains with RAPD and ISSR markers

Since the 1980s,Sargassum fusiforme has been cultivated in Zhejiang,South China,and nowadays it becomes one of the important commercial seaweeds in China.With traditions of eating habits in the East Asian countries,this brown alga is used as food,because it contained functional oligo/polysaccharides and chemical components,and was regarded playing roles in antioxidant activities and regulating immunology.Through over 15 years’selection,breeding and cultivation,we obtained three strains with good traits and testified their characters during the production,which included the cultivars with high yield and other two good characters,either all the selected strains were applied in the Sargassum production.To avoid confusion during the selection and nursery,it was preferred to establish one fingerprint for distinguishing the Sargassum cultivars from different strains.Random amplified polymorphic DNA(RAPD)and inter-simple sequence repeat(ISSR)methods were adopted to analyze the genetic diversities of the selected S.fusiforme strains.With that,one fingerprint with RAPD markers was constructed,and one sequence characterized amplifi ed region(SCAR)marker to S.fusiforme was obtained.It is indicated that the applied fingerprint could be valid in S.fusiforme genetic and germplasm justification,and will be positive to molecular marker assistance in its selection and cultivation.

Estimation of genetic parameters for upper thermal tolerance and growth-related traits in turbot Scophthalmus maximus using the Bayesian method based on Gibbs sampling

In order to carry out the genetic improvement of turbot upper thermal tolerance, it is necessary to estimate the genetic parameters of UTT（upper thermal tolerance） and growth-related traits. The objective of this study was to estimate genetic parameters for BW（body weight） and UTT in a two-generational turbot（Scophthalmus maximus L.） pedigree derived from four imported turbot stocks（England, France, Denmark and Norway）. A total of 42 families including 20 families from G1 generation and 22 families from G2 generation were used to test upper thermal tolerance（40–50 animals per family） in this study and the body weight of individuals were measured. The heritability of BW and UTT and the correlation between these two traits were estimated based on an individual animal model using Bayesian method based on two types of animal models with and without maternal effects.These results showed that the heritabilities for BW and UTT and phenotypic and genetic correlations between the two traits estimated from model without maternal effects were 0.239±0.141, 0.111±0.080, 0.075±0.026 and–0.019±0.011, respectively. The corresponding values from model with maternal effects were 0.203±0.115,0.055±0.026, 0.047±0.034 and –0.024±0.028, respectively. The maternal effects of BW and UTT were 0.050±0.017 and 0.013±0.004, respectively. The maternal effects had a certain influence on the genetic evaluation of the two traits. The findings of this paper provided the necessary background to determine the best selection strategy to be adopted in the genetic improvement program.

Characterization and differential expression of three GnRH forms during reproductive development in cultured turbot Schophthalmus maximus

Turbots( Schophthalmus maximus), one of the most important economic marine flatfish species, fail to undergo final spawning and spermiation naturally under artificial farming conditions. In vertebrates, reproduction is regulated by the brain-pituitary-gonadal axis(BPG-axis), and gonadotropin releasing hormone(GnRH) is one of its key components. Therefore, to better understand the physiology of reproduction in the turbot, three of the genes encoding GnRH subtypes— sbGnRH, c GnRH-II and sGnRH —were cloned and sequenced by isolating the cDNA sequences. The localizations and patterns of expression of their mRNAs were also evaluated during seasonal gonadal development. All three mRNAs were expressed abundantly in the brain; sbGnRH and sGnRH mRNAs were also detected in the gonads and pituitary gland, and sbGnRH expression was much higher than that of sGnRH, indicating the critical role of sbGnRH in regulating the BPG-axis. Moreover, the brain expression patterns of sbGnRH and sGnRH mRNAs showed an increased trend during gonadal development, peaking in mature stages. This indicated the direct regulation of gonadal development by the GnRH system. In addition, c GnRH-II mRNA expression showed no significant variations, suggesting that c GnRH-II is not critically involved in the control of reproduction. Further, the mRNA abundances of the three GnRH forms in the breeding season were significantly higher than those in immature and post-breeding stages in all analyzed brain areas. Therefore, we propose that sbGnRH is the most important hormone for the regulation of reproduction in turbot via the BPG-axis. These results will help in better understanding the reproductive endocrine mechanisms of turbots and lay the groundwork for additional studies aimed at comparing the reproductive physiology of wild individuals with those raised under artificial conditions.

Morphology and molecular phylogeny of Pleurosira nanjiensis sp. nov., a new marine benthic diatom from the Nanji Islands, China

A new marine benthic diatom, Pleurosira nanjiensis sp. nov., is described from the rocky intertidal zone of the Xiaochaiyu Island of the Nanji Islands in China. Its morphology was examined with light and scanning electron microscopy. Molecular phylogeny was reconstructed based on SSU rRNA and rbcL gene sequences. Pleurosira nanjiensis differs from congeners in possession of a combination of morphological features including the domed valve with broadly lanceolate, elliptical or circular valve outline, two elevated marginal ocelli, two （rarely three） rimportulae, and radiate striae.

Estimation of genetic parameters for upper thermal tolerances and growth-related traits in turbot Scophthalmus maximus

An abnormally high temperature produces a stress response in turbot causing large economic losses in the turbot aquaculture industry of China. A genetic improvement of the upper thermal tolerance (UTT) of turbot could allow cultured fi sh to adapt. A genetic evaluation of UTT is required for determining the practicability of including this trait into a breeding program. In this study, data were recorded from a temperature tolerance test conducted on 3 200 individual turbots from 32 full-sib groups. A cross-sectional linear model and a cross-sectional threshold probit model were used to analyze the test-period survival and a cross-sectional threshold logit model was used to analyze the test-day survival. In addition, phenotypic and genetic correlations between body weight and survival data were estimated. The estimated heritability values obtained from the cross-sectional linear model (CSL), the cross-sectional threshold (probit) model (THRp), and the cross-sectional threshold (logit) model (THRl) were 0.247 9±0.108 3, 0.288 3±0.161 2, and 0.106 9±0.045 2, respectively. The correlation coeffi cients among the full-sib family estimated breeding values (EBVs) obtained from the three models were greater than 0.998 6 and all models produced an almost identical family ranking. The accuracies of selection obtained with the CSL, THRp, and THRl model were 0.773 8, 0.775 4, and 0.784 4, respectively, the greatest from the THRl model. The genetic correlations between body weight and survival data EBVs from the CSL, THRp, and THRl models were 0.020 1,-6.201 1×10^-4 , and -3.115 4×10^-4 , respectively, and the phenotypic correlations between the two traits were -0.837 1 and -0.667 1, respectively. The findings of this study provide background information to determine the best strategy of selection for the genetic improvement of UTT in turbot.

Molecular characterization and expression profiles of four transformer-2 isoforms in the Chinese mitten crab Eriocheir sinensis

The transformer-2(tra-2) gene plays a key role in the regulatory hierarchy of sexual differentiation in somatic tissues and in the germline of Drosophila melanogaster.In this study,sequences and expression profiles of tra-2 in the Chinese mitten crab Eriocheir sinensis were characterized.Four tra-2 isoforms,designated as Estra-2a,Estra-2b,Estra-2c,and Estra-2d,were isolated.They all contained an RNA-recognition motif(RRM) and a linker region,which shared high similarity with other reported tra-2s.Sequence analysis revealed that Estra-2a,Estra-2b and Estra-2c are encoded by the same genomic locus and are generated by alternative splicing of the pre-mRNA.Compared with the other three isoforms,Estra-2d lacks the RS2 domain.Quantitative real-time PCR showed that all four isoforms were highly expressed in the fertilized egg,and in the 2-4 cell and blastula stages compared with larval stages(P<0.01),suggesting their maternal origin in early embryonic developmental stages.Notably,Estra-2a was highly expressed in male somatic tissues,while Estra-2c was significantly highly expressed in the ovary.These results suggest that Estra-2c is involved in sexual differentiation of the Chinese mitten crab.Our findings provide basic information for further functional studies of the tra-2 gene/protein in this species.

First record of Ulva californica in the mainland of China:a single alien parthenogenetic population in discontinuous distribution

Molecular investigations have raised concerns about the ecological risks of green tides caused by alien Ulva species in new habitats.The green tide-forming species U.californica Wille was generally considered to be native to North America,but new records have been widely reported in Europe,Asia,and Oceania in recent decades,indicating a strong dispersal capacity of the species.In this study,the first record of U.californica on the coastline of China's Mainland was reported,following a combined identification with multi-molecular markers and morphological characterization.It was shown that this species has a discontinuous distribution pattern along the coast of China's Mainland,with northern populations in the Yellow Sea and southern populations in the East China Sea and South China Sea.According to results of examination for life cycles and identification with mating type(MT)genetic markers,it was indicated that all U.californica samples were male gametophytes,and reproduced themselves through parthenogenesis solely.Combined with the fact that southern and northern populations are highly genetically identical,here we believed that U.californica was a recent alien species to China's Mainland with a rapid local spread.This finding provided evidences that the ability to reproduce in a variety of ways may play an important role in the spread of Ulva species,as well as essential basic data for marine risk management of green tides in China.In addition,according to the phylogeographic analysis,the possible geographical origin and global dispersal routes of U.californica were also proposed.

Transcriptome profiles of embryos before and after cleavage in Eriocheir sinensis:identification of developmental genes at the earliest stages

In crab, embryogenesis is a complicated developmental program marked by a series of critical events. RNA-Sequencing technology offers developmental biologists a way to identify many more developmental genes than ever before. Here, we present a comprehensive analysis of the transcriptomes of Eriocheir sinensis oosperms （Os） and embryos at the 2-4, cell stage （Cs）, which are separated by a cleavage event. Atotal of 18 923 unigenes were identified, and 403 genes matched with gene ontology （GO） terms related to developmental processes. In total, 432 differentially expressed genes （DEGs） were detected between the two stages. Nine DEGs were specifically expressed at only one stage. These DEGs may be relevant to stage-specific molecular events during development. A number of DEGs related to ＇hedgehog signaling pathway＇, ＇Wnt signaling pathway＇ ＇germplasm＇, ＇nervous system＇, ＇sensory perception＇ and ＇segment polarity＇ were identified as being up-regulated at the Cs stage. The results suggest that these embryonic developmental events begin before the early cleavage event in crabs, and that many of the genes expressed in the two transeriptomes might be maternal genes. Our study provides ample information for further research on the molecular mechanisms underlying crab development.

Near-infrared spectroscopy method for rapid proximate quantitative analysis of nutrient composition in Pacific oyster Crassostrea gigas

Glycogen,amino acids,fatty acids,and other nutrient components affect the flavor and nutritional quality of oysters.Methods based on near-infrared reflectance spectroscopy(NIRS)were developed to rapidly and proximately determine the nutrient content of the Pacific oyster Crassostreagigas.Samples of C.gigas from 19 costal sites were freeze-dried,ground,and scanned for spectral data collection using a Fourier transform NIR spectrometer(Thermo Fisher Scientific).NIRS models of glycogen and other nutrients were established using partial least squares,multiplication scattering correction first-order derivation,and Norris smoothing.The R_(C) values of the glycogen,fatty acids,amino acids,and taurine NIRS models were 0.9678,0.9312,0.9132,and 0.8928,respectively,and the residual prediction deviation(RPD)values of these components were 3.15,2.16,3.11,and 1.59,respectively,indicating a high correlation between the predicted and observed values,and that the models can be used in practice.The models were used to evaluate the nutrient compositions of 1278 oyster samples.Glycogen content was found to be positively correlated with fatty acids and negatively correlated with amino acids.The glycogen,amino acid,and taurine levels of C.gigas cultured in the subtidal and intertidal zones were also significantly different.This study suggests that C.gigas NIRS models can be a cost-effective alternative to traditional methods for the rapid and proximate analysis of various slaughter traits and may also contribute to future genetic and breeding-related studies in Pacific oysters.

Dynamic genetic analysis for body weight and main length ratio in turbot Scophthalmus maximus

The objective of this study was to estimate genetic parameters of body width(BW)to body length(BL)ratio(BW/BL)and of body weight traits(BWT)in turbot,and to elucidate the genetic mechanism of the two traits during ontogeny by dynamic genetic analysis.From 3 to 27 months,BW,BL and BWT of each communally stocked fish were measured every 3 months.The BW/BL ratio was measured at different sampling ages.A twotrait animal model was used for genetic evaluation of traits.The results showed that the heritability values of BW/BL ratio ranged from 0.2168 to 0.3148,corresponding to moderate heritability.The BWT heritability values ranged from 0.2702 to 0.3479 corresponding to moderate heritability.The heritability of BW/BL ratio was lower than that of BWT,except at 3 months of age.Genetic correlation between BW/BL ratio and BWT decreased throughout the measurement period.Genetic correlations were higher than the phenotypic correlations.The current results for estimating genetic parameters demonstrate that the BW/BL ratio could be used as a phenotypic marker of fast-growing turbot,and the BW/BL ratio and BWT could be improved simultaneously through selective breeding.

Eyes of diff ering colors in Alvinocaris longirostris from deep-sea chemosynthetic ecosystems: genetic and molecular evidence of its formation mechanism

Coloration is an important phenotypic trait for multiple adaptive functions.It is interesting to fi nd white-eye(AW)and orange-eye(AO)phenotypes in the shrimp Alvinocaris longirostris inhabiting the deep-sea cold seep and hydrothermal vent areas of the northwestern Pacifi c.By comparative transcriptome analyses,1491 diff erentially expressed genes(DEGs)were identified between AW and AO.Among them,many DEGs were associated with immunity,antioxidation,and detoxifi cation.Two signifi cant enzyme encoding genes,xanthine dehydrogenase,and tryptophan oxidase involved in pigment biosynthesis pathways were up-regulated in AW and AO,respectively,which might be related to the diff erences of white and orange eye phenotypes.Moreover,single nucleotide polymorphism(SNP)calling detected that genotypes of 28 SNP distributing in 14 unigenes were completely diff erent between AW and AO.Particularly,there were three and two non-synonymous mutations in immune genes crustin Pm5 and antimicrobial peptide,respectively.Results indicate that the diff erence in eye color is probably resulted from immune response to variable micro-environmental stressors encountered in the dispersal process of the shrimps,such as symbiotic microbes,pathogens,and toxic substances,and might be genetically fi xed at last.The suggested pathway preliminarily explained the formation mechanism of diff erent eye phenotypes in Alvinocaridid shrimps,providing a basis for further study on adaptive evolution of eyes in deep-sea chemosynthetic faunas.

Detection of two pathogenic marine ciliates Ancistrum haliotis and A. crassum(Ciliophora: Scuticociliatia) by fluorescence in situ hybridization

The scuticociliatid ciliates Ancistrum haliotis and A.crassum are parasites that may cause high mortality in the cultured abalone Haliotis spp.and the bivalve Ruditapes philippinarum.Traditional identification with silver staining methods is hampered by their morphological similarities to closely related species and the complicated procedures of morphological analysis.We designed two SSU rRNA-targeted oligonucleotide probes labeled with a fluorochrome,and optimized the fluorescence in situ hybridization(FISH)protocols for identification of A.halioti and A.crassum,respectively.The assays resulted in a clear identification by strong fluorescence signals from the oligonucleotide probes.The method can be used for quick and accurate quantitative analysis of A.haliotis and A.crassum infections on host molluscs.

Cryo-ET bridges the gap between cell biology and structural biophysics

Cryo-electron tomography（cryo-ET） is a cutting-edge technology providing three-dimensional in situ ultra-structural information of macromolecular machineries, organelles, and eukaryotic cells in their native environment at an unprecedented level of detail. Cryo-ET enables the direct observation of dynamic macromolecular architectures of bio-samples in their naturally occurring physiological state, without any harmful artifacts introduced by heavy metal staining, dehydration, and chemical fixation, which occur in traditional transmission electron microscopy. Over decades, cryo-ET has been providing insights into numerous aspects of cellular biology by revealing the pristinely preserved ultra-structures of different cellular components comprising the crowded and complex environment of the cell, thus, bridging the gap between cellular biology and structural biophysics. In this paper, we review the fundamentals of this technique, its recent advances in optics, detection devices, and computational algorithms. The enhancement of our understanding of structural cellular biology by combining these improvements, when integrated with other methods, such as cryo-focused ion beam milling,correlative light and electron microscopy, is discussed via a few examples from research groups worldwide. We also believe that cryo-ET applications in cell biology continue to provide fundamental insights into the field, revolutionizing structural biology itself.

Comparative chloroplast genomes of Ulva prolifera and U.linza(Ulvophyceae)provide genetic resources for the development of interspecific markers

The green seaweeds Ulva linza and U.prolifera are closely related species.They usually co-occur widely and have important ecological significance as primary producers thriving in the intertidal zone.In the Yellow Sea,a genetically unique floating ecotype of U.prolifera even bloomed to cause serious green tides.However,there is still a lack of appropriate molecular markers to distinguish these two species,partially due to limited evaluations on the intraspecific variations in U.prolifera among dif ferent ecotypes.Since organelle genomes could provide rich genetic resources for phylogenetic analysis and development of genetic markers,in this study,the chloroplast genome from one attached population of U.prolifera was completely sequenced,and comparative genomic analyses were performed with other existing chloroplast genomes from U.linza and the floating ecotype of U.prolifera.The results showed that in spite of the high level of collinearity among three genomes,there were plenty of genetic variations especially within the non-coding regions,including introns and gene spacer regions.A strategy was proposed that only those signals of variation,which were identical between two ecotypes of U.prolifera but divergent between U.linza and U.prolifera,were selected to develop the interspecific markers for U.linza and U.prolifera.Two candidate markers,psa B and pet B,were shown to be able to distinguish these two closely related species and were applicable to more attached populations of U.prolifera from a wide range of geographical sources.In addition to the interspecific marker,this study would also provide resources for the development of intraspecific markers for U.prolifera.These markers might contribute to the surveys for Ulva species composition and green tide monitoring especially in the Yellow Sea region.

Two new marine benthic diatom species of Hippodonta(Bacillariophyceae)from the coast of China:H.nanjiensis sp.nov.and H.qingdaoensis sp.nov.

The diversity and distribution of Hippodonta species on the coast of China are largely unknown due to limited investigations.In this paper,we report the detailed morphology and ultrastructure of two new species:H.nanjiensis sp.nov.,collected from intertidal sandy sediments of Nanji Islands,the East China Sea and H.qingdaoensis sp.nov.,collected from intertidal sandy sediments of the No.1 Bathing Beach in Qingdao City,the Yellow Sea.Both species occur mainly in the middle part of the intertidal zone.H.nanjiensis possesses a unique large round central area.H.qingdaoensis differs from congeners in having a unique stria pattern,wherein the areolae in the middle transapical striae are less developed or absent on the primary side of the valve,and two areolae per stria are apparent under light microscopy(LM).This study expands our knowledge of Hippodonta diversity and distribution on the coast of China.

Low genetic diversity in the endangered marine alga Silvetia siliquosa(Ochrophyta:Fucaceae)and the implication to conservation

Although signifi cant research eff orts have been targeted toward conservation and management of endangered terrestrial fl ora and fauna,attempts have been limited to conserve threatened seaweeds.Silvetia siliquosa is an ecologically and commercially vital brown alga that is uniquely distributed in the Yellow-Bohai Sea and along the southwest coast of Korea.A massive decline in its distribution range and biomass from the mid-1990s onward indicates that this species has become endangered.In the present study,we used nuclear internal transcribed spacer and concatenated mitochondrial cytochrome oxidase I subunit+intergenic spacer to estimate the genetic diversity,population connectivity,and degree of genetic diff erentiation of S.siliquosa in China and Korea.The molecular results exhibited strikingly low levels of haplotype/ribotype and nucleotide diversity in S.siliquosa populations,with only three mitochondrial haplotypes and nuclear ribotypes detected in 136 and 143 specimens,respectively.The analysis of molecular variance revealed 85%-95%of genetic variance among populations.Population diff erentiation coeffi cient(F_(ST))and gene fl ow(N_(m))suggested that two populations(JIN and GWA)along the southern coast of Korea are highly divergent from the others,with weak genetic exchange.No signifi cant genetic diff erentiation was observed among populations either in China or along the geographically proximate west coast of Korea.Thus,four independent management units were designated for sustainable management:the LII and RUS populations in China,the YEO and CHA populations along the west coast of Korea,and each of the GWA and JIN populations along the south coast of Korea.We suggest that artifi cial cultivation and transplantation of S.siliquosa are the eff ective approaches for restoration and conservation.