Sequence and phylogenetic analyses of the chloroplast 16S rRNA, tufA, and rbcL genes from Bryopsis hypnoides

Using shotgun sequencing data, the complete sequences of chloroplast 16S rRNA and tufA genes were acquired from native specimens of Bryopsis hypnoides (Qingdao, China). There are two group I introns in the 16S rRNA gene, which is structurally similar to that of Caulerpa sertularioides (Bryopsidales, Chlorophyta). The chloroplast-encoded tufA gene sequence is 1 230 bp long, very AT-rich (61.5%), and is similar to previously published 16S rRNA sequences of bryopsidinean algae. Phylogenetic analyses based on chloroplast 16S rRNA and tufA gene sequence data support previous hypotheses that the Bryopsidineae, Halimedineae, and Ostreobidineae are three distinct lineages. These results also confirmed the exclusion of Avrainvillea from the family Udoteaceae. Phylogenetic analyses inferred that the genus Bryopsis as sister to Derbesia; however, this clade lacked robust nodal support. Moreover, the phylogenetic tree inferred from rbcL GenBank sequences, combined with the geographical distributions of Bryopsis species, identified a strongly supportive clade for three differently distributed Asian Bryopsis species. The preliminary results suggesting that these organisms are of distinct regional endemism.     

Reproductive cycle of Ophiopholis mirabilis (Echinodermata: Ophiuroidea) in Zhangzi Island area,northern Yellow Sea

Ophiopholis mirabilis is a common species with a high population density on the western coasts of the northern Pacific Ocean.The number of O.mirabilis has been increasing recently in the scallop aquaculture zone(the Zhangzi Island area,northern Yellow Sea) in China.To explore the mechanism of its population variation,the reproductive cycle of O.mirabilis was investigated in this area(39°04'N;122°51'E) from February 2017 through January 2018 and determined by the monthly gonad index(GI),histological examinations of the gonads and the oocyte size-frequency distribution.O.mirabilis had a clear annual reproductive cycle that was synchronous between males and females.Sea temperature and food availability played important roles in O.mirabilis reproduction.The GI value was less reliable for determining reproductive activity in O.mirabilis because the nutritive tissues within the gonads may be utilized to synthesize gametes,leading to a decrease in GI during maturation.The histological results also show that abundant nutritive phagocytes were present in the gonads of O.mirabilis,which,together with the germ cells,affected the weight of the gonads.In addition,the mature oocytes of O.mirabilis were relatively small(75-150 μm),indicating that the larval development was planktotrophic.This study provided insights into the reproductive patterns and biology of O.mirabilis and is an essential basis for the quantity control of this species in aquaculture areas.     

Species identification and phylogenetic analysis of genus Nassarius(Nassariidae)based on mitochondrial genes

Genus Nassarius contains many subgenera, such as Zeuxis, Telasco, Niotha, Varicinassa, Plicarcularia, Nassarius s. str. and Reticunassa. On the basis of morphological characteristics of the shell and radula and sequences of mitochondrial cytochrome oxidase subunit I (COI) and 16S rRNA genes, Nassarius specimens collected from the South China Sea were identified and phylogenetically analyzed. Although Nassarius sp. and Nassarius (Varicinassa) variciferus were morphologically different in their shells, few variations were found among their radular teeth and sequences of mtCOI and mt16S RNA genes. Therefore, Nassarius sp. should be classified as N. (Varicinassa) variciferus. Nassarius (Zeuxis) sp. has only a subtle difference from Nassarius (Zeuxis) algidus on the shell, but it shows obvious differences in radular teeth and DNA sequence, indicating that they are two distinct species. Sequence divergence of mtCOI and mt16S RNA genes within Nassarius species was much lower than that between species, suggesting that these two genes are suitable for Nassarius species identification. Phylogenetic analysis (neighbor-joining and maximum parsimony) based on mtCOI and mt16S rRNA genes revealed the presence of two groups in genus Nassarius and a closest relationship between subgenera Zeuxis and Telasco. Species of subgenus Plicarcularia did not form a single clade. The molecular phylogeny was not congruent with the previous morphological phylogeny. The subgeneric divisions of genus Nassarius appear to be uncertain and unreliable.     

Morphology and molecular identification of Ulva forming green tides in Qingdao, China

Green tides are caused by the proliferation of chlorophytes under suitable hydrographic conditions.These blooms lead to environmental degradation and negatively impact the waters and seagrass beds,as well as fishing and other recreational activities in the bay.A comprehensive ecological understanding of the bloom dynamics,including the origin and persistence,is needed to foster management decisions.The algae in the great majority of green tide blooms usually belong to two genera of Ulvophyceae,Ulva and Enteromorpha.Ulva has been observed more often in recent years.In China,green tides occurred for the first time in the middle area of the Yellow Sea in 2007,and a large-scale algae blooming broke out in the middle and southern areas of the Yellow Sea in late May 2008.We identified them as Ulva prolifera by comparative analysis of the rDNA internal transcribed spacer 1 (ITS1),5.8S and ITS2 sequences in combination with microscopic observation.Morphological differences were found between the free-floating algae and the attached thalli.Various reproduction patterns of the free-floating algae include sexual,asexual and vegetative propagations,which played important roles in the long-term green tide persistence in China.The ITS sequences of the blooming algae were identical to those of the samples from the Lianyungang sea area but were different from the attached samples from the Qingdao sea area.The results infer that the blooms are originated from other sea areas rather than from the local attached populations.     

Two new records of Pyramidellidae (Gastropoda, Heterobranchia) from China seas

This paper reports two species of the family Pyramidellidae (Gastropoda, Heterobranchia), Otopleura nitida (A. Adams, 1854) and Rugadentia manzakiana (Yokoyama, 1922). The former was from the South China Sea and the latter was from the Yellow Sea and East China Sea. The study was based on the collections deposited in the Marine Biological Museum, Chinese Academy of Sciences. This is for the first time that these species have been recorded in China seas. In the current contribution, we describe and illustrate these two species, and compare them with similar species.     

Fouling acorn barnacles in China—a review

We review the species composition,distribution, and seasonal variation of fouling acorn barnacles in Chinese waters-from Bohai Sea and Yellow Sea to East and South China Seas. Thirty-two species of acorn barnacles were found, of which, the dominant species are Amphibalanus amphitrite, A. reticulatus, A. variegates, Balanus trigonus, Fistulobalanus kondakovi, Megabalanus tintinnabulum, Striatobalanus amaryllis, and Eurapha withersi in the fouling communities. A. amphitrite is the dominant species in the coastal waters of Bohai Sea and Yellow Sea and A. reticulatus is dominant in the East and South China Seas. The settlement period of fouling acorn barnacles is usually in summer and autumn. From north to south with the decrease of latitude, their settlement period obviously extends, even to the whole year, and the species number also increases.0ther environmental factors, such as salinity and distance from shore, also play an important role in the distribution of fouling acorn barnacles.     

Taxonomic and genetic status of lancelet in Weihai coastal waters based on mitochondrial DNA sequence

Lancelets (subphylum Cephalochordata) are a transitional species between invertebrates and vertebrates. They are currently listed in the Second Order of Protected Animals in China. Lancelets were first documented in the waters around the city of Weihai (Shandong, China) in 2002. However, little is known about the phylogeny of this population. We analyzed the sequences of cytochrome b (Cyt b) and cytochrome oxidase c subunit I (CO I) genes from samples collected from coastal waters in the cities of Weihai and Qingdao (~150 km to the south). We analyzed 176 sequences, of which 150 were novel sequences and 26 were obtained from GenBank. Our results suggest that (1) lancelets in the two cities belong to the species Branchiostoma japonicus and have a high level of genetic diversity; (2) there is a high level of gene flow and low level of genetic differentiation between lancelets from the two cities; (3) demographic expansion occurred an estimated 1.1 million years (Ma) ago (mid Pleistocene) for lancelets in Weihai-Qingdao; and (4) the divergence between B. belcheri and B. japonicus was estimated at between 37.75 Ma (early Oligocene)-46.5 Ma (late Eocene).     

A new species and new recorded species of Lumbrineridae Schmarda, 1861 (Annelida: Polychaeta) from China

Based on material deposited in the Marine Biological Museum of the Chinese Academy of Sciences(Qingdao),a new species,Lumbrineris sinensis sp.nov.,as well as Augeneria albidentata(Ehlers,1908),which is recorded for the first time from coastal water of China,are reported in the present paper.The specimens examined were collected during 1958 to 1960 from the Yellow Sea,East China Sea and South China Sea at depths of 4 to 182 m.     

Oncholaimus zhangi sp. nov. (Oncholaimidae,Nematoda) from the intertidal zone of the East China Sea

A new species of free-living marine nematode is described from intertidal sediment of the East China Sea.The new species belongs to the Oncholaimidae family.Oncholaimus zhangi sp.nov.is characterized by males having a large ventral swelling situated just posterior to the middle of the tail,no precloacal papilla,and a row of 6–7 ventral genital setae.The tails of both males and females are similar,being conico-cylindrical in shape.The new species is distinguished from other closely related species by the presence of a prominent ventral swelling at the tail of males and a similar tail shape in both sexes.About 110valid species in this genus have been recorded to date.     

Genetic variation in two sea cucumber （Apostichopus japonicus） stocks revealed by ISSR markers

Sea cucumber Apostichopus japonicus samples were collected in Changdao, Penglai (PL), 27 individuals, and Lingshandao, Qingdao (QD), 30 individuals, in the Shandong Peninsula, China. Ten SSR primers were used to assess the genetic variation and relationship between and within the two stocks. Respectively, for each stock, the percentage of polymorphic bands was 85.2% and 86.9%; the gene diver- sity was 0.360 5 and 0.342 8; and the Shannon’s information index was 0.515 0 and 0.499 0. At species level, the percentage of polymorphic bands was 92.2%, the total gene diversity was 0.378 9 and the Shannon’s information index was 0.550 4. The coefficient of overall genetic differentiation and the ge- netic distances between the stocks were also calculated to be 0.073 0 and 0.079 6 using the POPGENE program. Results show that the genetic diversity of the two stocks is still large but the genetic distance between the two stocks is close. A dendrogram was constructed for the 57 individuals from the two stocks, showing that the genetic structure was unitary for PL stock but complex for QD stock.     

Seasonal variation in community structure and body length of dominant copepods around artificial reefs in Xiaoshi Island, China

This study aims to investigate the seasonal variations in copepod community structure and prosome length of dominant species from March 2009 to January 2010 around artificial reefs in Xiaoshi Island,Yellow Sea,Weihai,China.Samples were collected using two types of plankton net(Model I and Model II) for different-sized copepods.The number of taxon was calculated from the data of both the net types,while the copepod abundance was done using the samples from Model II only.Sixteen species of planktonic copepods,including 5 dominant species,were recorded.Results reveal that Oithona similis was the first dominant species from March to June,and was replaced by Paracalanus parvus in September;both dominated the copepod community in January.Acartia hongi was the second dominant species from March to September.Centropages abdominalis was the third dominant species from March to June,and was replaced by O.similis in September and Corycaeus affinis in January.C.affinis was the fourth dominant species in September.Population density of the dominant copepods was compared with that of other similar regions.We found that the dominant species were mostly small copepods(<1 mm) except for adult Centrapages abdominalis.Seasonal variation in prosome length of O.similis,C.abdominalis,and C.affinis,and their copepodites were studied for the first time in China.For P.parvus and A.hongi,seasonal trends in prosome length variation were similar with those in Jiaozhou Bay,Yellow Sea,Qingdao,China,in a similar temperate domain.The results are helpful for future calculation of copepod biomass and production,and for investigation of the relationship between copepods and fish resources.     

A Preliminary Phylogenetic Analysis ofLuidia （Paxillosida： Luidiidae） from Chinese Waters with Cytochrome Oxidase Subunit I （COI） Sequences

Luidia Forbes （Paxillosida： Luidiidae） are common soft bottom sea stars with 49 described species. Because of substan- tial morphological diversity, the taxonomy of the genus is complex and hasn＇t been resolved definitely. In order to resolve general taxonomic issues, and determine species boundaries and phylogenetic relationships within the genus Luidia, the sequences of tyro- chrome oxidase subunit I （COI） gene from 24 specimens of Luidia, belonging to eight taxa in Chinese waters, were studied. Three sequences of two species in genus Luidia from GenBank were used to analyze the phylogenetie relationships. The molecular phy- logeny exhibited three main clades, each with strong bootstrap support： Clade A including Luidia quinaria from the Sea of Japan; Clade B including seven nominal species （L. quinaria von Martens, L. yesoensis Goto, L. ehangi Liu, Liao and Li, L. orientalis Fisher, L. avicularia Fisher, L. longispina Sladen and L. hardwicki Gray） from Chinese waters; and Clade C including L. maculata Miiller ＆ Troschel from Chinese waters. Our molecular phylogeny results support the morphological Quinaria-Group and Alter- nata-Group assigned by D6derlein. Seven nominal species we sampled do not exhibit genetic distances that are large enough to rec- ognize them as separate species. Cryptic species may exist in ＇Luidia quinaria＇ from the Yellow Sea and the Sea of Japan. Meaning- ful morphological characters need further investigation in Luidia.     

Genetic diversity in two Japanese flounder populations from China seas inferred using microsatellite markers and COI sequences

Japanese flounder is one of the most important commercial species in China; however, information on the genetic background of natural populations in China seas is scarce. The lack of genetic data has hampered fishery management and aquaculture development programs for this species. In the present study, we have analyzed the genetic diversity in natural populations of Japanese flounder sampled from the Yellow Sea (Qingdao population, QD) and East China Sea (Zhoushan population, ZS) using 10 polymorphic microsatellite loci and cytochrome c oxidase subunit I (COI) sequencing data. A total of 68 different alleles were observed over 10 microsatellite loci. The total number of alleles per locus ranged from 2 to 9, and the number of genotypes per locus ranged from 3 to 45. The observed heterozygosity and expected heterozygosity in QD were 0.733 and 0.779, respectively, and in ZS the heterozygosity values were 0.708 and 0.783, respectively. Significant departures from Hardy-Weinberg equilibrium were observed in 7 of the 10 microsatellite loci in each of the two populations. The COI sequencing analysis revealed 25 polymorphic sites and 15 haplotypes in the two populations. The haplotype diversity and nucleotide diversity in the QD population were 0.746±0.072 8 and 0.003 34±0.001 03 respectively, and in ZS population the genetic diversity values were 0.712±0.047 0 and 0.003 18±0.000 49, respectively. The microsatellite data (F_st =0.048 7, P<0.001) and mitochondrial DNA data (F_st =0.128, P<0.001) both revealed significant genetic differentiation between the two populations. The information on the genetic variation and differentiation in Japanese flounder obtained in this study could be used to set up suitable guidelines for the management and conservation of this species, as well as for managing artificial selection programs. In future studies, more geographically diverse stocks should be used to obtain a deeper understanding of the population structure of Japanese flounder in the China seas and adjacent regions.     

Fauna and standing crop of freshwater mussels in Poyang Lake, China

Freshwater mussels are a major component of the zoobenthos and they therefore play an important role in freshwater ecosystems. The community composition and the standing crop of freshwater mussels in Poyang Lake, the largest freshwater lake in China and one of two lakes connected to the Changjiang (Yangtze) River, were investigated from February 2008 to May 2009. Mussels were sampled in hydrologically and geographically distinct sampling areas. A total of 42 species and 13 genera were identified, of which 32 species were endemic to China. Two species, Unio douglasiae and Lamprotula caveata, dominated all sampling areas. The number of genera and species, and the number of species within each genus, differed significantly among sampling areas. Freshwater mussels were very abundant, with an overall density of 0.28 ± 0.22 ind./m2 and biomass of 4.08 ± 3.96 g/m2. Significant variation observed among sampling areas and among genera was probably due to the variety of habitats among the areas sampled and to the adaptability of different freshwater mussels to environments. This study suggests that Poyang Lake and other freshwater lakes in the region are important sites of freshwater mussel biodiversity and conservation.     

A PRELIMINARY ANALYSIS ON THE CHARACTERISTICS OF THE KUROSHIO FRONTAL EDDY IN THE EAST CHINA SEA IN SPRING

The Kuroshi'o front eddy's surface and sectional isothermal distribution characteristics were analyzed on the basis of observation data obtained in April 13-16 of 1989 in the East China Sea. It was found from the similarity between these isothermal distributions with those in January and beginning of June for the years 1986-1990 that the Kuroshio front eddy often occurred from March to the beginning of June. The Kuroshio front eddy movement in the East China Sea in spring was along two routes: the Okinawa Trough route, and the continental shelf slope route. The two moving routes both in the surface layer and in the section are described, their causes are discussed, and differences are compared.