Genetic diversity and population structure of three dominant myctophid fishes (<Emphasis Type="Italic">Diaphus theta,Stenobrachius leucopsarus</Emphasis>, and <Emphasis Type="Italic">S. nannochir</Emphasis>) in the North Pacific Ocean

We compared the genetic diversity of three dominant myctophid fishes in the North Pacific Ocean that have different diel vertical migration patterns on the basis of the nucleotide sequences of the mitochondrial gene for cytochrome b. No genetic structure was detected for each of these three species. The genetic diversity progressively increased for Diaphus theta, a diel migrant species showing clear diel vertical migration; Stenobrachius leucopsarus, a semi-diel migrant, in which only part of the population migrates vertically; S. nannochir, a non-diel migrant. All three species were suggested to have experienced a recent, sudden population expansion. Interspecific differences in genetic diversity might be attributable to differences in the degree of population size reduction during the glacial periods; this degree in turn corresponds to the energy demand of the fishes.     

Vertical Distributions of Larvae of the Clam <Emphasis Type="Italic">Ruditapes philippinarum</Emphasis> and the Striped Horse Mussel <Emphasis Type="Italic">Musculista senhousia</Emphasis> in Eastern Ariake Bay,Southern Japan

We have examined the vertical distributions of planktonic bivalve larvae, particularly the clam Ruditapes philippinarum and the mussel Musculista senhousia which are common and abundant on tidal flats of eastern Ariake Bay, southern Japan. Submersible pumping gear was used to take samples at 2 stations every 2 hours during the spring tide over a whole tidal cycle and/or through daytime and nighttime. Water samples were pumped up from 3 to 5 depths from the surface to sea bottom. Regardless of tidal cycles, D-shaped larvae were concentrated near the surface, while umbo larvae were found at the surface to intermediate depths. On average, these larval densities were significantly higher at the station close to the shore than the offshore station, with no significant difference between daytime and nighttime and between flood and ebb tides at each station. The velocity and direction of water movement at both stations revealed remarkable difference between the surface and bottom waters. The larvae at the surface may quickly disperse and be transported elsewhere, while those in water close to the sea bottom may tend to be retained.     

利用线粒体DNA 控制区部分序列分析不同地理群体大泷六线鱼遗传多样性

利用PCR技本扩增了6个野生地理群体大泷六线鱼(Hexagrammos otakii)线粒体DNA控制区的部分序列,共获得352bp碱基序列,31尾个体定义了22种单倍型.中性检验Fu's Fs值为-0.14881(P＜0.01).分子变异分析(AMOVA)结果显示:遗传分化指数Fst=0.7398(P＜0.01),73.98％的变异来自群体间,26.02％的变异来自群体内.根据不同个体间的遗传距离构建了NJ和MP系统树,两种方法获得了相似的进化树.其中青岛群体与旅顺群体的一部分个体单独成支,其它群体聚为1支.研究结果表明,大泷六线鱼群体遗传多样性较为丰富,群体间发生了较大的遗传分化,证明线粒体DNA控制区基因可用于大泷六线鱼群体内及群体间遗传多样性的分析.     

Morphological differences between three groups of <Emphasis Type="Italic">Luciogobius guttatus</Emphasis> on the Korean Peninsula

Three ecological and geographical forms of Luciogobius guttatus from the western and eastern coasts of the Korean Peninsula as well as the coasts of, Jeju Island which is located to the south of the peninsula are described and compared based on morphological characteristics. The environments of these three coastal areas are different, and the morphometrics show a clear pattern of differentiation and reveal the discreteness of the western, eastern and Jeju groups. In addition, a discriminant analysis (DA), a principal component analysis (PCA) of morphometric characteristics, and a t-test for meristic characteristics all show that the three populations of L. guttatus differ from each other. Three divergent lineages of L. guttatus are described with the possibility of niche differentiation between the western, eastern and Jeju coastal habitats. The standard length of eastern populations was greater than that of western and Jeju populations, and those with morphometric characteristics had a longer body. In addition, eastern populations had a higher number of vertebrae. We speculate that the characteristics of eastern populations evolved through the difference in tide level and gravel density, whereas the difference between western and Jeju populations is due to geographical distance.     

基于线粒体控制区的中国东南沿海黄鳍棘鲷群体遗传多样性和遗传分化

本研究通过对线粒体控制区序列进行分析的技术手段,对中国宁德（ND）、厦门（XM）、漳浦（ZP）、南澳岛（NAD）、湛江（ZJ）、海口（HK）与北海（BH）的7个野生黄鳍棘鲷（Acanthopagrus latus）群体的遗传多样性和遗传分化进行比较分析。结果表明：中国东南沿海的野生黄鳍棘鲷群体呈现出中等以上的遗传多样性特征,48个黄鳍棘鲷样品的控制区序列长度为580 bp,单倍型数目为44个,总体的单倍型多样性指数（Hd）和核苷酸多样性指数（Pi）分别为0.996和0.017 30。7个野生黄鳍棘鲷群体间的遗传分化程度较低,各群体间的遗传距离为0.010 28~0.029 87。群体间的遗传分化系数（FST）为-0.068 41~0.545 86,北海群体和其他群体间存在一定程度的遗传分化。AMOVA分析显示群体内的遗传变异占比为83.34%,大部分遗传变异发生在组群内;但北海群体与其他6个群体分为两个组群后,组群间的遗传变异占比为42.66%,两个组群间存在较大程度的遗传变异。基于Kimrua距离法构建的系统进化树中显示北海群体聚为一个分支,另外6个群体聚为一支。基于TCS network构建的单倍型网络图中44个单倍型混杂在一起,未按照水系格局和地理距离进行分布,表明黄鳍棘鲷群体间未形成多个单系群。本研究可为中国东南沿海野生黄鳍棘鲷群体的种质资源利用和保护提供科学依据。     

Intercellular localization of nitrogenase in a non-heterocystous cyanobacterium (cyanophyte), <Emphasis Type="Italic">Trichodesmium</Emphasis> sp. NIBB1067

The nitrogenase activities of cyanobacteria of the genus Trichodesmium are strictly light-dependent, although they do not develop heterocysts. Recently, the development of heterocyst-equivalent cells (diazocytes) was suggested in Trichodesmium spp. However, no cells with a similar appearance to diazocytes could be found in Trichodesmium sp. NIBB1067. An immunocytochemical analysis of nitrogenase in individual cells was performed using polyclonal antibodies generated from DNA fragments of genes encoding the Fe-protein and the α-and the β-subunit of the MoFe-protein of nitrogenase of Trichodesmium sp. NIBB1067. Visualization of the antibody binding was carried out using a horseradish peroxidase-conjugated secondary antibody and a chromogenic substrate, 3,3′-diaminobenzidine, to avoid the masking effects caused by the bright autofluorescence emitted by Trichodesmium cells. Nitrogenase proteins were detected in almost all the cells (always higher than 95% of total cells) grown under nitrogen-limited conditions. These results indicate that Trichodesmium sp. NIBB1067 does not differentiate heterocyst-equivalent cells.     

Molecular phylogeny of the genus <Emphasis Type="Italic">Chondracanthus</Emphasis> (Rhodophyta), focusing on the resurrection of <Emphasis Type="Italic">C. okamurae</Emphasis> and the description of <Emphasis Type="Italic">C. cincinnus</Emphasis> sp. nov.

Determining the taxonomic status of the red algal genus Chondracanthus based on morphological characters is challenging due to the similarity and high degree of plasticity of the thallus. Since the taxonomic history of several Chondracanthus species remains unclear, we analyzed the plastid rbcL and mitochondrial COI genes of the specimens from Korea and Japan, in combination with morphological observations, to examine their phylogenetic relationships. Our results confirmed the distinction of C. okamurae, which is separated from C. intermedius, and identified a novel species, C. cincinnus sp. nov. Three species (C. okamurae, C. intermedius and C. cincinnus) formed a monophyletic clade with C. tenellus. C. okamurae is distinguished by linear, narrow, cylindrical to compressed, slightly recurved axes, and a high-intertidal to subtidal distribution. It was collected from Korea and Japan, while C. intermedius was identified from Japan only. A new species, Chondracanthus cincinnus sp. nov., is characterized by linear, compressed, strongly recurved axes, and a low-intertidal to subtidal distiribution. Based on the molecular phylogeny using rbcL and COI data, we herein resurrect C. okamurae as a distinct species and identify C. cincinnus as a new species.     

Crown foliage dynamics of mangrove <Emphasis Type="Italic">Kandelia obovata</Emphasis> in Manko Wetland,Okinawa Island,Japan

The crown foliage dynamics of Kandelia obovata (S., L.) Yong were investigated, including leaf recruitment, survival and leaf growth. Newly flushed leaves occurred successively throughout the year, with a maximum in July and a minimum in January. The highest leaf death was in August, whereas it was the lowest in January. The relative mortality rate of leaves seemed to depend on the season when new leaves flushed. The growth pattern of leaves varied among seasons. The maximum leaf area was significantly larger in winter than in autumn, but showed no significant difference between the winter and the other seasons. The half-expansion period and the intrinsic rate of increase were respectively longer and lower in winter than in the other seasons. Therefore, leaves flushed in summer grew faster in their initial stage and attained their maximum leaf area sooner than those flushed in winter. This most likely results from the difference in temperature between summer and winter. The crown leaf area was almost stable throughout the year, as if homeostatic control is likely to function. Stipule litterfall decreased significantly with increasing flower and propagule litterfalls, indicating that a high production of flowers and propagules results in low leaf recruitment. This may be because flower production and propagule growth impose a severe burden on K. obovata.     

Habitat suitability index of Chub mackerel (<Emphasis Type="Italic">Scomber japonicus</Emphasis>) from July to September in the East China Sea

The habitat quality of Chub mackerel (Scomber japonicus) in the East China Sea has been a subject of concern in the last 10 years due to large fluctuations in annual catches of this stock. For example, the Chinese light-purse seine fishery recorded 84000 tons in 1999 compared to 17000 tons in 2006. The fluctuations have been attributed to variability in habitat quality. The habitat suitability Index (HSI) has been widely used to describe fish habitat quality and in fishing ground forecasting. In this paper we use catch data and satellite derived environmental variables to determine habitat suitability indices for Chub mackerel during July to September in the East China Sea. More than 90% of the total catch was found to come from the areas with sea surface temperature of 28.0°–29.4°C, sea surface salinity of 33.6–34.2 psu, chlorophyll-a concentration of 0.15–0.50 mg/m³ and sea surface height anomaly of −0.1–1.1 m. Of the four conventional models of HSI, the Arithmetic Mean Model (AMM) was found to be most suitable according to Akaike Information Criterion analysis. Based on the estimation of AMM in 2004, the monthly HSIs in the waters of 123°–125°E and 27°30′–28°00′ N were more than 0.6 during July to September, which coincides with the catch distribution in the same time period. This implies that AMM can yield a reliable prediction of the Chub mackerel’s habitat in the East China Sea.     

Production of <Emphasis Type="Italic">Acartia steueri</Emphasis> (Copepoda: Calanoida) in Ilkwang Bay,Southeastern Coast of Korea

Production of the marine calanoid copepod Acartia steueri was measured from 2 October 1991 to 8 October 1992 at a station in Ilkwang Bay, on the southeastern coast of Korea. Phytoplankton standing stock ranged over 1.0 to 9.3 mg chl.a m⁻³, and annual primary productivity (by the C-14 method) at three stations was estimated at 200 gC m⁻² yr⁻¹. Acartia steueri (nauplii + copepodids + adults) were present in the plankton throughout the year, with seasonal variation in abundance. Biomass of A. steueri, excluding the NI stage, was 0.01–4.55 mgC m⁻³ (mean: 0.68 mgC m⁻³) with peaks in November, February, May and July-early August, and relatively low biomass in September– January. Instantaneous growth rates of the nauplius stages were higher than the copepodid stages. Annual production of A. steueri was 25.1 mgC m⁻³ yr⁻¹ (or 166 mgC m⁻² yr⁻¹), showing peaks in November, May and July–August with a small peak in February, and low production in December–April and September–October. There were no significant relationships between the daily production rate of A. steueri and temperature or chlorophyll a concentration, indicating that unknown other factors might be related to the variation of the production rate.     

Degradation of outer membrane proteins of <Emphasis Type="Italic">Synechococcus</Emphasis> sp. <Emphasis Type="Italic">in vitro</Emphasis> and <Emphasis Type="Italic">in situ</Emphasis>

To examine the possibility that outer membrane proteins (OMP) of Synechococcus sp. remain in seawater, we investigated the stability of OMPs in vitro and in situ. Some fractions prepared from Synechococcus sp. CSIRO-94 were treated with trypsin and proteinase K. Four tightly bound OMPs were separated from Synechococcus. We designated the two major OMPs of 52 kDa and 48 kDa as Omp52Sy and Omp48Sy, respectively. Degradation of the OMP in natural seawater was monitored in microcosms to which intact Synechococcus cells and outer membrane (OM) were added. Omp52Sy and Omp48Sy were the most stable against trypsin and proteinase K among the OMPs when they were embedded in the OM. However, in the microcosm experiment using intact cells, Omp52Sy and Omp48Sy were detected in the particulate fraction only during the first 4 days, after which they could not longer be detected. Omp52Sy and Omp48Sy were the most stable proteins among the Synechococcus OMPs in vitro, but they might be degraded in situ. This indicates that stability of Synechococcus porin differs depending on complex formation with other membrane molecules, which might cause different preservation of microbial membrane proteins in the dissolved protein pool in the ocean. This study suggests that Gram negative bacterial OM with thin peptidoglycan forms a lipid bilayer that proptects OMP, but Synechococcus OM with thick peptidoglycan cannot form a lipid bilayer. The incomplete bilayer might not be able to protect from protease attack in the natural environment.     

Detection of nitrogenase in individual cells of a natural population of <Emphasis Type="Italic">trichodesmium</Emphasis> using immunocytochemical methods for fluorescent cells

The intercellular distribution of nitrogenase was detected in a natural population of Trichodesmium spp. collected from Shitaba Bay, Uwajima, Ehime Prefecture, Japan. Polyclonal antibodies raised against the Fe-protein and the MoFe-protein (α-subunit) of nitrogenase were used as probes. Visualization of the antibody binding to the nitrogenase was performed using horseradish peroxidase-conjugated secondary antibody and the chromogenic substrate, 3,3′-diaminobenzidine tetrahydrochloride, to avoid the masking effects caused by the bright autofluorescence emitted by the Trichodesmium cells. Nitrogenase proteins were detected in 66 to 81% of the trichomes. More than 77% of the cells were immunostained in individual nitrogenase-containing trichome. The data show natural populations of Trichodesmium spp. do not developed heterocyst-equivalent cells.     

New polymorphic microsatellite markers derived from hemocyte cDNA library of Manila clam <Emphasis Type="Italic">Ruditapes philippinarum</Emphasis> challenged by the protozoan parasite <Emphasis Type="Italic">Perkinsus olseni</Emphasis>

Manila clam Ruditapes philippinarum is one of the most important benthic animals in the coastal north Pacific region, where clam populations have been mixed genetically through trade and aquaculture activities. Accordingly, identification of the genetically different clam populations has become one of the most important issues to manage interbreeding of the local and introduced clam populations. To identify genetically different populations of clam populations, we developed 11 expressed sequence tag (EST)-microsatellite loci (i.e., simple sequence repeat, SSR) from 1,128 clam hemocyte cDNA clones challenged by the protozoan parasite Perkinsus olseni. Genotype analysis using the markers developed in this study demonstrated that clams from a tidal flat on the west coast contained 6 to 19 alleles per locus, and a population from Jeju Island had 4 to 20 alleles per locus. The expected heterozygosity of the 2 clam populations ranged from 0.472 to 0.919 for clams from the west coast, and 0.494 to 0.919 for clams from Jeju Island, respectively. Among the 11 loci discovered in this study, 7 loci significantly deviated from the Hardy-Weinberg equilibrium after Bonferroni correction. The 5 loci developed in this study also successfully amplified the SSRs of R. variegatus, a clam species taxonomically very close to R. philippinarum, from Hong Kong and Jeju Island. We believe that the 11 novel polymorphic SSR developed in this study can be utilized successfully in Manila clam genetic diversity analysis, as well as in genetic discrimination of different clam populations.     

New records of three <Emphasis Type="Italic">Paraprionospio</Emphasis> species (Polychaeta: Spionidae) from Korean waters

The spionid polychaete Paraprionospio pinnata (Ehlers 1901) has been widely reported from Korean waters. We examined some specimens belonging to the genus Paraprionospio that had been collected from Korean waters, and identified them as Paraprionospio coora Wilson, 1990, Paraprionospio cordifolia Yokoyama, 2007 and Paraprionospio patiens Yokoyama, 2007, which are new to the fauna of Korea. The present study suggests misidentification of the specimens that have been previously reported as P. pinnata from Korean waters.     

<Emphasis Type="Italic">In situ</Emphasis> Enclosure Experiment Using a Benthic Chamber System to Assess the Effect of High Concentration of CO<Subscript>2</Subscript> on Deep-Sea Benthic Communities

In order to evaluate the environmental impact associated with sequestration of carbon dioxide in the deep sea, a free fall type field experimental device, the benthic chamber, was developed. In situ experiments to expose deep-sea communities to elevated concentrations of carbon dioxide (average of 20,000 ppm, 5,000 ppm and control) were carried out using this device 3 times, viz., in the winter of 2002 and in the spring and the summer of 2003, in the Kumano Trough at a depth of 2,000 m. In the long-term experiments (about two weeks in winter of 2002 and summer of 2003), the abundance of meiobenthos declined whereas that of bacteria increased under the condition of 20,000 ppm carbon dioxide compared with the control. Among meiofauna, the abundance of foraminifers at the same concentration of carbon dioxide became less than the control even in the short-term (3 days in spring of 2003) experiment, suggesting that organisms with a calcium carbonate exoskeleton are more sensitive to the raised concentration of carbon dioxide. The respiration rate of the benthic community exposed to 20,000 ppm was lower in the early stage of the experiment than in the latter half, whereas it was opposite under the condition of 5,000 ppm. The increase of biological activity in the 20,000 ppm exposure group is probably due to an increase of bacteria adapted to high carbon dioxide concentrations. The present results suggest that the influence of carbon dioxide on the deep-sea benthic ecosystem does not follow a simple, linear relationship with concentration.     

Encystment and excystment of <Emphasis Type="Italic">Gyrodinium instriatum</Emphasis> Freudenthal et Lee

In the present study, we have investigated the conditions influencing encystment and excystment in the dinoflagellate Gyrodinium instriatum under laboratory conditions. We incubated G. instriatum in modified whole SWM-3 culture medium and in versions of modified SWM-3 from which NO₃ ⁻, PO₄ ³⁻, NO₃ ⁻ + PO₄ ³⁻, or Si had been omitted and observed encystment. Percentage encystment was high in the media without N and without P, while the percentage encystment in the medium lacking both N and P was highest. Moreover, to investigate N or P concentration which induced the encystment, Gyrodinium instriatum was also incubated in media with different concentrations of inorganic N and P; the concentrations of NO₂ ⁻ + NO₃ ⁻ and PO₄ ³⁻ were measured over time. The precursors of cysts appeared within 2 or 3 days of a decrease in NO₂ ⁻ + NO₃ ⁻ or PO₄ ³⁻ concentration to values lower than 1 μM or 0.2 μM, respectively. When cysts produced in the laboratory were incubated, we observed excystment after 8–37 days, without a mandatory period of darkness or low temperature. We incubated cysts collected from nature at different temperatures or in the dark or light and observed excystments. Natural cysts excysted at temperatures from 10 to 30°C, in both light and dark, but excystment was delayed at low temperatures. These studies indicate that G. instriatum encysts in low N or P concentration and excysts over a wide temperature range, regardless of light conditions, after short dormancy periods.     

<Emphasis Type="Italic">Dracograllus trukensis</Emphasis> sp. nov. (Draconematidae: Nematoda) from a seagrass bed (<Emphasis Type="Italic">Zostera</Emphasis> spp.) in Chuuk Islands,Micronesia, Central Western Pacific Ocean

A new species of free-living marine draconematid nematode, Dracograllus trukensis sp. nov., is described based on the specimens collected from the sediments of a intertidal seagrass bed from Chuuk Islands, Micronesia. Dracograllus trukensis sp. nov. differs from other species of the genus by the combination of the following characteristics: the presence of numerous minute spiny ornamented body cuticular annules in both sexes, eight cephalic adhesion tubes inserted on the head capsule in both sexes, the presence of stiff posteriorly directed setae anterior to posterior adhesion tubes in both sexes, the shape (large, elongated, open loop-shaped in male and large, elongated, closed loop-shaped in female) and position (longer ventral arm extending to the first body annule in male) of amphideal fovea, shorter spicule length (34–42 μm), the presence of sexual dimorphism in shape and length of the non-annulated tail terminus, and number of posterior sublateral adhesion tubes (10 in male and 13–15 in female) and posterior subventral adhesion tubes (8–10 in male and 9–11 in female). A comparative table on the biogeographical and ecological characteristics of the species of Dracograllus is presented. This is the first taxonomic report on the genus Dracograllus from Chuuk Islands, Micronesia, central western Pacific Ocean.     

Morphological identification of <Emphasis Type="Italic">Alexandrium</Emphasis> species (Dinophyceae) from Jinhae-Masan Bay,Korea

Outbreaks of paralytic shellfish poisoning (PSP) and dense blooms caused by Alexandrium species in Jinhae-Masan Bay, Korea have been nearly annual events for many years. However, excluding some Alexandrium species responsible for PSP, there are no critical reports on the morphology of Alexandrium species in this bay. To identify the Alexandrium species based on detailed morphological features, vegetative cells collected water samples and established by the incubation of resting cysts isolated from sediment trap samples were analyzed. Four species of Alexandrium were identified: Alexandrium affine, A. fundyense, A. catenella, and A. insuetum. Morphological features of these species were basically consistent with those outlined in previous studies. However, the ventral pore and the connecting pore on the sulcal plate, which have been accepted as diagnostic characteristics for the identification of A. fundyense and A. catenella, need to be reevaluated, indicating that useful morphological features for identifying these two species should be recommended to avoid confusion in the classification of species in genus Alexandrium.     

Age and Growth of <Emphasis Type="Italic">Sillago aeolus</Emphasis> in Okinawa Island,Japan

In a study of the age and growth of Sillago aeolus, a total of 414 fish were collected from December 2000 to March 2002, around Okinawa Island. Ages of 403 specimens were determined by otoliths. Monthly changes in the percentage of otoliths with an opaque zone in the outer margin indicated that an opaque ring formed once a year, occurring between February and May. The rings can therefore be defined as annual rings. Ages of females were estimated to be 0+ to 4+, while those of males were 0+ to 2+. The von Bertalanffy growth curves were well fitted to age-standard length data of males and females, as follows: Male: L_t = 209.6 [1 − exp{−0.70 (t + 0.58)}]. Female: L_t = 297.7 [1 − exp{−0.42 (t + 0.61)}].     

Spawning characteristics of <Emphasis Type="Italic">Clupea pallasii</Emphasis> in the coastal waters off Gyeongnam,Korea, during spawning season

The purpose of this study is to research the characteristics of spawning grounds of Clupea pallasii found at 1 spawning ground located in the coast of Yeongun-ri, Tongyeong, Gyeongnam, and 3 stations in Jinhae Bay. Diving observation was performed from January to June, 2014 at the coast of Yeongun-ri, and from February to June, 2016 at Jinhae Bay after total 4 stations (A: Gusan-myeon, B: Haengamdong, C: Hacheong-myeon, D: Jam-do) were constituted. During observation period, 1 spawning ground was found in Tongyeong area in January. In Jinhae Bay spawning grounds were found at 3 stations (A, B, D) out of 4 stations in February. Regarding the surrounding environment of spawning ground found at Yeongunri, Tongyeong, various kinds of seaweeds were distributed up to 1–5 m depth, and many branch type red algae were distributed. In case of Jinhae Bay, seaweeds were widely distributed from shallow water to 5 m depth. In addition, the scope that eggs were attached was considerably wide compared to Tongyeong area, and they were found at all the 3 spots, so C. pallasii is thought to use Jinhae Bay for its spawning ground widely. Eggs were attached from the surface to 3–4 m, and like Tongyeong area, eggs were attached to mostly branch type of red algae. The results of this study suggest that the spawning season of the C. pallasii in coast waters off Gyeongnam is until mid-February.