Kofoidinium, Spatulodinium and other kofoidiniaceans (Noctilucales, Dinophyceae) in the Pacific Ocean

Examples of rarely reported dinoflagellates of the family Kofoidiniaceae F.J.R. Taylor (Noctilucales) from the northwest, equatorial and southeast Pacific Ocean are described and illustrated. Kofoidinium was the most ubiquitous genus with a maximum abundance of 10 cells L(-1). Specimens of this genus were identified to four species: Kofoidinium sp. that showed a pointed extension that emerges from the antero-ventral region and K. velelloides, both of which had diameters that ranged from 40 to 200 microm; Kofoidinium pavillardii which showed a rounded epitheca and a larger size (approximately 300-700 microm in diameter); and another species, tentatively identified as K. splendens, that contained red circular inclusions. Further research is needed to clarify the characteristics that separate K. splendens from the other species. This study is the first to record the genus Spatulodinium in tropical waters and in the southern hemisphere. S. cf. pseudonoctiluca was found in the southeast Pacific Ocean, as well as other smaller specimens with a different shape or disposition of the tentacle that may belong to two other species. In the northwest and equatorial Pacific, specimens of Spatulodinium showed a green pigmentation that suggested the existence of the first species known in the order Noctilucales to contain its own chloroplasts. Immature stages of kofoidiniaceans, some containing symbiotic microalgae, are illustrated, as well as mature stages related to Pomatodinium and to unknown genera of kofoidiniaceans. Kofoidiniaceans are shown to be common and widely distributed in the Pacific, and are probably also frequent in other oceans, but are rarely recognised.     

The impact of the 2003 summer heat wave and the 2005 late cold wave on the phytoplankton in the north-eastern English Channel

The phytoplankton composition was investigated at two fixed stations in the north-eastern English Channel from November 1997 to December 2005. The warmest temperatures in European historical records were recorded in August 2003. This event was associated with an exceptional abundance peak of the dinoflagellates Akashiwo sanguinea (9600 cells L(-1)) and Ceratium fusus. The lowest February temperatures for the 1998-2005 period were recorded in 2005, coinciding with the absence, for the first time in recent decades, of the spring bloom of Phaeocystis globosa. The 'de-eutrophication', mainly the reduction of river nutrient loads, is progressively reducing the magnitude of the Phaeocystis blooms. Exceptionally in 2005, the colder temperatures increased water column mixing, favouring the dominance of tychoplanktonic diatoms until early March (pre-bloom period). The delay in spring stratification, lower light availability due to turbidity (resuspended sediment) and organic matter, and competition with tychoplanktonic diatoms contributed to retard the timing of the spring phytoplankton bloom and disadvantage the development of Phaeocystis. The summer 2003 European heat wave is expected to have had little influence on total annual primary production, because it occurred at mid-summer, the period of lowest annual phytoplankton abundance. However, the anomalous weather in the second half of winter 2005 did affect the annual primary production.     

Changes in bacterial populations during red tides caused by Mesodinium rubrum and Gymnodinium catenatum in North West Coast of Spain

Heterotrophic bacterial communities associated with four red tides caused by Mesodinium rubrum and Gymnodinium catenatum in two Galician Rias (North West Spain) were examined. Three of these were produced by the Mesodinium rubrum and the causative organism of a toxic bloom was Gymnodinium catenatum . In early stages of all the blooms, the diversity decreased but the total marine bacterial counts increased by one or two logs. Vibrio numbers were also incremented by two logs in two blooms of M. rubrum , while in the other bloom of this organism and in the red tide caused by G. catenatum a decrease in number of these bacteria was observed. A total of 116 bacterial strains were identified at the genus level and grouped into 12 phena. During the decomposition processes of two blooms of M. rubrum a zooplanktonic-type bacterial succession was observed ( Vibrio , pseudomonads and Moraxella-Acinetobacter ). On the other hand, during decomposition of the other red tide of M. rubrum and the bloom of G. catenatum , a typical phytoplanktonic-type succession occurred, as Pseudomonas and Moraxella groups became dominant for all the process. These results support the conflicting taxonomical position of M. rubrum . After the blooms, the changes in the community point towards the restablishment of the normal bacterial flora of the estuary (increase in diversity and decreases of bacterial numbers). Only the Vibrio strains, isolated from the non-toxic first and second red tides, displayed cytotoxic activities. A relationship among bacterial cytotoxicity and toxic effects of blooms cannot therefore be established.     

PARASITISM OF PHOTOSYNTHETIC DINOFLAGELLATES BY THREE STRAINS OF AMOEBOPHRYA (DINOPHYTA): PARASITE SURVIVAL,INFECTIVITY, GENERATION TIME,AND HOST SPECIFICITY1

Amoebophrya ceratii (Koeppen) Cachon is an obligate parasite of dinoflagellates and may represent a species complex. However, little is known about the biology and host range of different strains of Amoebophrya Cachon. Here, we determined parasite generation time and dinospore infectivity, survival, and ability to infect nonprimary hosts for strains of Amoebophrya from Akashiwo sanguinea (Hirasaka) G. Hansen et Moestrup, Gymnodinium instriatum (Freudenthal et Lee) Coats comb. nov., and Karlodinium micrum (Leadbeater et Dodge) J. Larsen. Akashiwo sanguinea was readily infected, with parasite prevalence reaching 100% in dinospore:host inoculations above a 10:1 ratio. Parasitism also approached 100% in G. instriatum, but only when inoculations exceeded a 40:1 ratio. Karlodinium micrum appeared partially resistant to infection, as parasite prevalence saturated at 92%. Parasite generation time differed markedly among Amoebophrya strains. Survival and infectivity of dinospores decreased over time, with strains from G. instriatum and A. sanguinea unable to initiate infections after 2 and 5 days, respectively. By contrast, dinospores from Amoebophrya parasitizing K. micrum remained infective for up to 11 days. Akashiwo sanguinea and G. instriatum were not infected when exposed to dinospores from nonprimary Amoebophrya strains. Karlodinium micrum, however, was attacked by dinospores of Amoebophrya from the other two host species, but infections failed to reach maturity. Observed differences in host–parasite biology support the hypothesis that Amoebophrya ceratii represents a complex of host‐specific species. Results also suggest that Amoebophrya strains have evolved somewhat divergent survival strategies that may encompass sexuality, heterotrophy during the “free‐living” dinospore stage, and dormancy.     

ULTRASTRUCTURE OF GYMNODINIUM AUREOLUM (DINOPHYCEAE): TOWARD A FURTHER REDEFINITION OF GYMNODINIUM SENSU STRICTO

Examination of the ultrastucture of the unarmored dinoflagellate Gymnodinium aureolum (Hulburt) G. Hansen (syn: Gyrodinium aureolum Hulburt) revealed the presence of nuclear chambers, which are specialized differentiations of the nuclear envelope, similar to those described in the type species of Gymnodinium , G. fuscum (Ehrenberg) Stein and certain other Gymnodinium species. The nuclear pores were restricted to these chambers. In the flagellar apparatus a nuclear fibrous connective linked the longitudinal microtubular root and the nucleus. This structure had so far been observed only in Gymnodinium spp. and in the heterotrophic species Actiniscus pentasterias (Ehrenberg) Ehrenberg, Nematodinium armatum (Dogiel) Kofoid et Swezy and Polykrikos kofoidii Chatton. Another unusual feature of G. aureolum was the presence of a striated fiber in the longitudinal flagellum, a feature previously only found in Ceratium furca (Ehrenberg) Claparède et Lachmann and C. tripos (O.F. Müller) Nitzsch. Gymnodinium aureolum also possessed a prominent ventral protrusion associated with the peduncle and containing electron opaque material. It is concluded that G. aureolum belongs to the Gymnodinium sensu stricto group. This may be a temporary classification, however, because G. aureolum and its allies differ from the type species G. fuscum by the presence of a transverse striated root, striated collars, trichocysts, and a peduncle.     

Reconstructing the history of an invasion: the toxic phytoplankton species <Emphasis Type="Italic">Gymnodinium catenatum</Emphasis> in the Northeast Atlantic

The phytoplankton species Gymnodinium catenatum is responsible for major worldwide losses in aquaculture due to shellfish toxicity. On the West coast of the Iberian Peninsula, toxic blooms have been reported since the mid-1970s. While the recent geographical spread of this species into Australasia has been attributed to human-mediated introduction, its origin in the Northeast Atlantic is still under debate. Gymnodinium catenatum forms a highly resistant resting stage (cyst) that can be preserved in coastal sediments, building-up an historical record of the species. Similar cyst types (termed microreticulate) are produced by other non-toxic Gymnodinium species that often co-occur with G. catenatum. We analysed the cyst record of microreticulate species in dated sediment cores from the West Iberian shelf covering the past ca. 150 years. Three distinct morphotypes were identified on the basis of cyst diameter and paracingulum reticulation. These were attributed to G. catenatum (35.6–53.3 μm), G. nolleri (23.1–36.4 μm), and G. microreticulatum (20.5–34.3 μm). Our results indicate that G. catenatum is new to the NE Atlantic, where it appeared by 1,889 ± 10, expanding northwards along the West Iberian coast. The earliest record is from the southernmost sample, while in the central Portuguese shelf the species appears in sediments dated to 1,933 ± 3, and in the North, off Oporto, in 1,951 ± 4. On the basis of the cyst record and toxic bloom reports, we reconstruct the invasive pathway of G. catenatum in the NE Atlantic. Although human-mediated introduction cannot be discarded, the available evidence points towards natural range expansion, possibly from NW Africa.     

Changes in bacterial populations during red tides caused by Mesodinium rubrum and Gymnodinium catenatum in North West Coast of Spain

Heterotrophic bacterial communities associated with four red tides caused by Mesodinium rubrum and Gymnodinium catenatum in two Galician Rias (North West Spain) were examined. Three of these were produced by the Mesodinium rubrum and the causative organism of a toxic bloom was Gymnodinium catenatum. In early stages of all the blooms, the diversity decreased but the total marine bacterial counts increased by one or two logs. Vibrio numbers were also incremented by two logs in two blooms of M. rubrum, while in the other bloom of this organism and in the red tide caused by G. catenatum a decrease in number of these bacteria was observed. A total of 116 bacterial strains were identified at the genus level and grouped into 12 phena. During the decomposition processes of two blooms of M. rubrum a zooplanktonic-type bacterial succession was observed (Vibrio, pseudomonads and Moraxella-Acinetobacter). On the other hand, during decomposition of the other red tide of M. rubrum and the bloom of G. catenatum, a typical phytoplanktonic-type succession occurred, as Pseudomonas and Moraxella groups became dominant for all the process. These results support the conflicting taxonomical position of M. rubrum. After the blooms, the changes in the community point towards the restablishment of the normal bacterial flora of the estuary (increase in diversity and decreases of bacterial numbers). Only the Vibrio strains, isolated from the non-toxic first and second red tides, displayed cytotoxic activities. A relationship among bacterial cytotoxicity and toxic effects of blooms cannot therefore be established.     

Gymnodinium catenatum Graham isolated from the Portuguese coast: Toxin content and genetic characterization

The bloom forming marine dinoflagellate Gymnodinium catenatum Graham has been linked to paralytic shellfish poisoning (PSP) outbreaks in humans. Along the Portuguese coast (NE Atlantic), G. catenatum shows a complex bloom pattern, raising questions about the origin and affinities of each bloom population. In this work, the variability within six cultured strains of G. catenatum isolated from Portuguese coastal waters (S coast, W coast and NW coast), between 1999 and 2011, was investigated. The strains were analyzed for toxin profiling and intra-specific genetic diversity. Regarding the toxin profile, differences recorded between strains could not be assigned to the time of isolation or geographical origin. The parameter that most influenced the toxin profile was the life-cycle stage that originated the culture: vegetative cell versus hypnozygote (resting cyst). At the genetic level, all strains showed similar sequences for the D1–D2 region of the large subunit (LSU) of the nuclear ribosomal DNA (rDNA) and shared complete identity with strains from Spain, Algeria, China and Australia. Conversely, we did not find a total identity match for the ITS-5.8S nuclear rDNA fragment. After sequence analysis, two guanine/adenine (R) single nucleotide polymorphisms (SNP 1 and 2) were detected for all strains, in the ITS1 region. This species has been reported to present very conservative LSU and ITS-5.8S rDNA regions, though with few SNP, including SNP1 of this study, already attributed to strains from certain locations. The SNP here described characterize G. catenatum populations from Portuguese waters and may represent valuable genetic markers for studies on the phylogeography of this species.     

GYMNODINIUM COROLLARIUM SP. NOV. (DINOPHYCEAE)—A NEW COLD‐WATER DINOFLAGELLATE RESPONSIBLE FOR CYST SEDIMENTATION EVENTS IN THE BALTIC SEA1

A naked dinoflagellate with a unique arrangement of chloroplasts in the center of the cell was isolated from the northern Baltic proper during a spring dinoflagellate bloom (March 2005). Morphological, ultrastructural, and molecular analyses revealed this dinoflagellate to be undescribed and belonging to the genus Gymnodinium F. Stein. Gymnodinium corollarium A. M. Sundström, Kremp et Daugbjerg sp. nov. possesses features typical of Gymnodinium sensu stricto, such as nuclear chambers and an apical groove running in a counterclockwise direction around the apex. Phylogenetic analyses based on partial nuclear‐encoded LSU rDNA sequences place the species in close proximity to G. aureolum, but significant genetic distance, together with distinct morphological features, such as the position of chloroplasts, clearly justifies separation from this species. Temperature and salinity experiments revealed a preference of G. corollarium for low salinities and temperatures, confirming it to be a cold‐water species well adapted to the brackish water conditions in the Baltic Sea. At nitrogen‐deplete conditions, G. corollarium cultures produced small, slightly oval cysts resembling a previously unidentified cyst type commonly found in sediment trap samples collected from the northern and central open Baltic Sea. Based on LSU rDNA comparison, these cysts were assigned to G. corollarium. The cysts have been observed in many parts of the Baltic Sea, indicating the ecologic versatility of the species and its importance for the Baltic ecosystem.     

Dynamics of morphological and anatomical changes in leaf tissues of an interspecific hybrid of oil palm during acquisition and development of somatic embryogenesis

Oil palm is an economically important plant species due to its high oil production per unit area. Large-scale clonal propagation of the species’s elite specimens is only possible through somatic embryogenesis, although methodology is partially still unknown and insufficiently understood. Current study characterizes in morphological and anatomical terms the acquisition and development stages of somatic embryogenesis of the oil palm’s immature leaves. The respective embryogenic stages were analyzed and characterized: immature leaves (initial explants); leaves with calli formation; leaves which failed to respond to calli formation; leaves with formation of root structures; primary calli; primary calli with differentiation of embryogenic calli; embryogenic calli; pro-embryogenic calli; calli with differentiated somatic embryos; somatic embryos at globular and torpedo stage; and mature fruit zygotic embryos. Cell masses emerged after approximately 60 days of cultivation through the proliferation of cells associated to initial explants´ vascular bundles. Consequently, the formation of two different types of calli was identified, namely, primary and embryogenic, respectively consisting partially and completely of meristematic cell clusters. After 420 days of cultivation, the propagules formed somatic embryos with no connection to source tissues, initially composed (globular stage) of a very characteristic ground meristem and protoderm. After 480 days of cultivation, as the cultures matured (torpedo stage), procambial strands, a structural characteristic also observed in mature zygotic embryos, were reported. The results provide an in-depth understanding of somatic embryogenesis of immature leaves of oil palm. Further, current analysis develops morphological markers at different stages of development obtained during the process.     

Balechina and the new genus Cucumeridinium gen. nov. (Dinophyceae), unarmored dinoflagellates with thick cell coverings

The genus Balechina (=subgenus Pachydinium) was established for heterotrophic gymnodinioid dinoflagellates with a thick cell covering. The type species, B. pachydermata (=Gymnodinium pachyderm‐atum), showed numerous fine longitudinal striae, whereas B. coerulea (=G. coeruleum) showed ~24 prominent longitudinal surface ridges or furrows and a distinctive blue pigmentation. We have investigated the morphology and molecular phylogeny of these taxa and the species Gymnodinium cucumis, G. lira and G. amphora from the western Mediterranean, Brazil and Japan. Sudden contractions at the cingulum level were seen in B. pachydermata, which also showed a high morphological variability which included morphotypes that have been described as Amphidinium vasculum, G. amphora, G. dogielii and G. gracile sensu Kofoid and Swezy. Molecular phylogeny based on small subunit rRNA gene sequences revealed that Balechina coerulea, G. cucumis and G. lira formed a clade distantly related to the clade of the type species, B. pachydermata, and G. amphora. We propose the new genus Cucumeridinium for the species with longitudinal ridges and a circular apical groove (Cucumeridinium coeruleum comb. nov., C. lira comb. nov. and C. cucumis comb. nov.), and Gymnodinium canus and G. costatum are considered synonyms of C. coeruleum. The genus Balechina remains for the species with a double‐layer cell covering, bossed surface with fine striae, and an elongated elliptical apical groove. At present, the genus is monotypic containing only B. pachydermata.     

果实种类及发育阶段对梨小食心虫幼虫钻蛀率和蛹重的影响

【目的】明确梨小食心虫Grapholitha molesta在寄主果实上的钻蛀率和蛹重与寄主种类和果实发育阶段的关系,为其田间预测预报和综合防控提供技术依据。【方法】在室内条件下测定了接卵后72 h梨小食心虫初孵幼虫在不同发育阶段的3个种共6个品种水果［苹果Malus pumila (嘎啦苹果), 桃Amygdalus persica (大久保毛桃、丰白毛桃和霞光油桃), 梨Pyrus bretschneideri (砀山酥梨和玉露香梨)］果实上的钻蛀率以及在不同果实状态的4个水果品种(大久保毛桃、丰白毛桃、砀山酥梨和玉露香梨)果实上72 h的钻蛀率;比较了接卵后24 h和72 h初孵幼虫在不同发育阶段的3个苹果品种(嘎啦苹果、富士苹果和金冠苹果)果实上的钻蛀率;并测定了不同发育阶段的金冠苹果、富士苹果、嘎啦苹果、砀山酥梨和丰白毛桃上的梨小食心虫蛹重。【结果】接卵后72 h梨小食心虫初孵幼虫在6个不同水果品种果实上的钻蛀率从高到低依次为：霞光油桃＞嘎啦苹果＞砀山酥梨＞玉露香梨＞大久保毛桃＞丰白毛桃;除霞光油桃外,其他5个水果品种果实的发育阶段显著影响初孵幼虫钻蛀率。对于嘎啦苹果,接卵后72 h初孵幼虫在其幼果上的钻蛀率最高(73.69%),在其着色期果实上钻蛀率最低(32.51%);在大久保毛桃、丰白毛桃、砀山酥梨和玉露香梨上,接卵后72 h初孵幼虫的钻蛀率均随果实生长发育而增加。初孵幼虫在3 个苹果品种着色期和成熟期果实上的钻蛀率随处理时间的增加而显著下降,而在其幼果上变化不显著。寄主种类及其发育阶段也显著影响梨小食心虫蛹重,成熟毛桃和梨饲养的雌、雄蛹重显著高于未成熟果实饲养的蛹重,着色期苹果不利于蛹重的增加。【结论】果实种类及发育阶段显著影响梨小食心虫初孵幼虫钻蛀率和蛹重,取食成熟期梨和桃的梨小食心虫初孵幼虫钻蛀率和蛹重显著高于取食未成熟果实的个体。     

Dynamics of spicule production in the marine sponge <Emphasis Type="Italic">Hymeniacidon perlevis</Emphasis> during in vitro cell culture and seasonal development in the field

To characterize the formation of silica spicules, the dynamics of spiculogenesis of an intertidal marine sponge Hymeniacidon perlevis (Montagu 1818) (Porifera: Demospongiae) were investigated by measuring the gene expression of silicatein (the enzyme responsible for spicule silicification) and the dimensional changes of spicules during the developmental process of individual sponges and in cell cultures of primmorphs of archaeocyte-dominant cell populations. The different developmental stages of spicules were documented by time-lapse microscopy and observed by transmission electron microscopy during a 1-month culture period. During its annual life cycle, H. perlevis has four different developmental stages: dormancy, resuscitation, bloom, and decline. Field-grown individual sponge samples at different stages were collected over 7 months (March to September 2005). The dimensions of the silica spicules from these samples were microscopically measured and statistically analyzed. This analysis and the material properties of the spicules allowed them to be classified into four groups representing the different developmental stages of spiculogenesis. Silicatein expression in the bloom stage was more than 100 times higher than that in the other stages and was correlated with the spicule developmental stage. The trend of spicule formation in field-grown sponges was consistent with the trend in cell culture. A new parameter, the maturation degree (MD) of spicules (defined as the ratio of actual to theoretical silica deposition of mature spicules), was introduced to quantify spicule development. Silica spiculogenesis during H. perlevis development was delineated by comparing MD and silicatein expression.     

棕囊藻北部湾株的18S rDNA分子鉴定

为研究北部湾棕囊藻(Phaeocystis)藻华的成因,采用PCR克隆了棕囊藻北部湾株核糖体18S r DNA序列。结果表明,棕囊藻北部湾株具有游动单细胞与群体结构两种形态;其18S r DNA序列和NCBI基因库中球形棕囊藻(Phaeocystis globosa)的同源性为99%~100%,在系统进化树上与不同海域来源的球形棕囊藻聚在一大分支上,且与球形棕囊藻间的遗传距离均小于其他种。首次从分子生物学上确定棕囊藻北部湾株为球形棕囊藻。     

Embryo development in vitro of cat oocytes cryopreserved at different maturation stages

The purpose of this study was to evaluate the ability of cat oocytes, at different stages of maturation, to survive after cryopreservation and to assess their subsequent development following IVM and IVF. In the initial toxicity trial, immature oocytes were exposed to different concentrations of DMSO and ethylene glycol (EG). Resumption of meiosis and metaphase II were evaluated after removal of the cryoprotectant and IVM. The highest rates of resumption of meiosis (51.4%) were achieved after exposure to 1.5 mol l(-1) of cryoprotectants, and no difference was observed with control oocytes. Metaphase II was obtained in 25.7% (P<0.01) and 22.9% (P<0.005) of oocytes exposed to 1.5 mol l(-1) of DMSO and ethylene glycol, although at lower rates than in control oocytes (54.4%). On the basis of this finding, 1.5 mol l(-1) of cryoprotectant was chosen for freezing cat oocytes at the germinal vesicle stage (immature) or at metaphase II stage (mature). Post-thaw viability was assessed by the evaluation of the embryo development in vitro. After fertilization, mature oocytes frozen in ethylene glycol cleaved in better proportions (38.7%) than immature oocytes (6.8%, P<0.001), and no differences were observed in the cleavage rate of oocytes frozen at different maturation stages with DMSO (immature 12.8%; mature 14.1%). Embryonic development beyond the 8-cell stage was obtained only when mature oocytes were frozen with ethylene glycol (11.3%). This study suggests that cryopreserved cat oocytes can be fertilized successfully and that their development in vitro is enhanced when mature oocytes are frozen with ethylene glycol. The stage of maturation may be a key element in improving cat oocyte cryopreservation.     

Litter Overlap in Gambusia hubbsi: Superfetation Revisited

The frequency of litter overlap, the simultaneous presence of two litters at different stages of development within a single female, varies among populations of Gambusia hubbsi in different habitats on Andros, Bahamas. In freshwater bluehole and shallow water populations, less than 2% of the females carried two litters, independent of the difference in developmental stage between the litters. In well field populations, 2% of the females carried two litters composed of immature and mature oocytes, whereas 10% carried developing oocytes and late stage embryos, and 16% carried mature oocytes (or older) and late stage embryos. Among females from populations that we introduced into unoccupied well fields 2% carried two litters composed of immature and mature oocytes, about 14% carried developing oocytes and late stage embryos, and less than 6% carried mature oocytes (or older) and late stage embryos. When two litters composed developing oocytes and late stage embryos were present, the two litters were of equal size, and the total number of offspring was 1.5 times larger than expected on the basis of female body length. When two litters composed mature oocytes (or older stages) and late stage embryos were present, the two litters were also of equal size, and the total number of offspring was as expected on the basis of female body length. Litter overlap was independent of female size. A comparison of the consequences of litter overlap for strictly lecithotrophic and strictly matrotrophic modes of reproduction with the patterns of litter overlap observed in G. hubbsi, suggests that litter overlap in G. hubbsi reduces the cost of reproduction, but does not increase the rate of offspring production.     

Vernal sedimentation trends in north Norwegian fjords: temporary anomaly in 234Th particulate fluxes related to Phaeocystis pouchetii proliferation

We report data of a naturally occurring radionuclide, ²³⁴Th, an in situ tracer, to investigate vertical export of biogenic matter during a vernal bloom of Phaeocystis pouchetii in the fjords of northern Norway. To optimise sampling of different stages of the bloom, three fjords with increasing oceanic influence (Balsfjord, Malangen fjord and Ullsfjord, respectively) were investigated in April 1997. Contrasting situations were encountered between the three fjords: the proliferation of P. pouchetii in Ullsfjord surface waters coincided with a drastic reduction of particulate ²³⁴Th fluxes in traps, although particulate organic carbon (POC) and dimethylsulphoniopropionate (DMSP) were exported and ²³⁴Th was available in surface waters. When large colonies make up a significant fraction of the vertical flux, as observed in Ullsfjord in April 1997, there may be a large and rapid change in the POC/²³⁴Th ratio, further complicating the use of ²³⁴Th as a tracer for POC export. The results suggest that the proliferation of Phaeocystis pouchetii during vernal bloom could temporary increase OC/²³⁴Th ratio of particles and delay the particulate export of ²³⁴Th, and probably of other particle-reactive species, from surface waters.     

Research Note: Molecular phylogenetic affiliations of Dissodinium pseudolunula, Pheopolykrikos hartmannii, Polykrikos cf. schwartzii and Polykrikos kofoidii to Gymnodinium sensu stricto species (Dinophyceae)

Partial large subunit ribosomal DNA (LSU rDNA) sequences of Dissodinium pseudolunula Swift ex Elbrächter et Drebes, Pheopolykrikos hartmannii (Zimmermann) Matsuoka et Fukuyo, Polykrikos cf. schwartzii Bütschli and Polykrikos kofoidii Chatton were analyzed to reveal their phylogenetic status. The four athecate dinoflagellate species fell within the clade of Gymnodinium sensu G. Hansen et Moestrup supported by high likelihood values without apparent phylogenetic relationships to neither Gymnodinium species nor each other. Their genetic affiliations to typical Gymnodinium species were morphologically supported by the loop-shaped apical grooves in Pheopolykrikos and Polykrikos species and the Gymnodinium -like motile stage at least once during pleomorphic life cycles in D. pseudolunula and Ph. hartmannii .     

MOLECULAR EVIDENCE IDENTIFIES BLOOM-FORMING PHAEOCYSTIS (PRYMNESIOPHYTA) FROM COASTAL WATERS OF SOUTHEAST CHINA AS PHAEOCYSTIS GLOBOSA

Chen, Y. Q., Zhou, H. & Qu, L. H. Key Laboratory of Gene Engineering of Education Ministry, Biotechnology Research Center, Zhongshan University, Guangzhou 510275, P. R. China Sequence data from the 18S small subunit ribosomal RNA gene and rDNA ITS regions have been used to identify the species of a Phaeocystis (Prymnesiophyta) that caused harmful algae blooms in the coastal waters of southeast China. This Phaeocystis has morphological and physiological features that differ from those previously described for either P. globosa Scherf- fel or P. pouchetii (Hariot)Lagerheim. However, the sequence comparison of the Phaeocystis 18S rDNA and rDNA ITS clearly showed that it was remarkably similar to several isolates of P. globosa. Thus, the species isolated from the southeast coast of China is identified as P. globosa rather than P. cf. pouchetii or another species. Our results also demonstrate that phenotypes of different members of the genus Phaeocystis are variable, apparently changing in response to environmental conditions. It is concluded that, on the basis of this phylogenetic analysis, the bloom forming southeast China coast species of Phaeocystis most likely originated from an endemic warm-water, rather than a foreign source.     

Molecular cloning and expression of TRα and TRβ in the protandrous cinnamon clownfish,Amphiprion melanopus during sex reversal

We divided the process of sex reversal from immature male to mature female in the protandrous cinnamon clownfish (Amphiprion melanopus) into six developmental stages as follows: I, immature male; II, mature male; III, male at 60 days after female removal; IV, male at 90 days after female removal; V, male at 120 days after female removal; and VI, mature female. Thyroid hormone receptors α (TRα) and β (TRβ) cDNAs were cloned from the ovary and mRNA expression levels were compared during the sex-reversal process. The nucleotide sequences of the TRα and TRβ cDNA were 1230 and 1188 bp in length with open reading frames encoding peptides of 409 and 395 amino acids, respectively. We observed that TRα mRNA and protein levels were high in all stages except the immature gonad, while TRβ mRNA levels were higher in the mature ovary than in any other gonadal stage. We injected gonadotropin-releasing hormone analogue to identify its effects on TRs mRNA in immature fish. The mRNA levels of TRs increased significantly. We therefore propose that TRs are related to testicular development as well as ovarian development in cinnamon clownfish. The present study also provides basic data on the role of TRs during sex reversal in fish.