Cyst‐Theca Relationship and Phylogenetic Position of Impagidinium caspienense Incubated from Caspian Sea Surface Sediments: Relation to Gonyaulax baltica and Evidence for Heterospory within Gonyaulacoid Dinoflagellates

We investigate the cyst‐theca relationship of Impagidinium caspienense. Through an incubation experiment, we succeeded in examining the motile stage. Additional molecular analysis of single‐cyst PCR (LSU and SSU rDNA) reveal that the cyst is related to the species Gonyaulax baltica Ellegaard et al. ( 2002 ). The ability of this species to belong to two types of cyst‐based genera (spiniferate and impagidinioid) suggests that environmental (particularly salinity) and not genetic factors explain the formation of both morphotypes by G. baltica, which provides evidence for heterospory in this species. The affiliation to G. baltica demonstrates that I. caspienense is not endemic to the Caspian Sea. The phylogenetic position of several other gonyaulacoid species is also documented: Impagidinium pallidum, Ataxiodinium choane, Pyxidinopsis psilata, Spiniferites belerius, and Spiniferites ramosus. The LSU and SSU rDNA based phylogenies suggest that the genera Impagidinium and Spiniferites are not monophyletic, and that P. psilata and A. choane are close to Gonyaulax verior and Gonyaulax polygramma, respectively. In addition, this study accentuates the importance of cyst morphology in the classification of the Gonyaulacales.     

Relationship between the dinoflagellate cyst Spiniferites pachydermus and Gonyaulax ellegaardiae sp. nov. from Izmir Bay,Turkey, and molecular characterization

Here, we established the cyst‐motile stage relation‐ship for Spiniferites pachydermus through incubation of cysts with a characteristically microreticulate/perforate surface isolated from Izmir Bay in the eastern Aegean Sea of the eastern Mediterranean. The morphology of the motile stage was similar to Gonyaulax spinifera but had a different size, overhang, displacement and reticulations. Based on the distinct morphology of the cyst and morphological differences in motile cells, we assigned S. pachydermus from Izmir Bay to the new species Gonyaulax ellegaardiae. We elucidate the phylogenetic relationship of G. ellegaardiae through large and small subunit ribosomal DNA and show that it forms a clade with other species that belong to the G. spinifera complex.     

PHYLOGENETIC POSITION OF CRUSTOMASTIX STIGMATICA SP. NOV. AND DOLICHOMASTIX TENUILEPIS IN RELATION TO THE MAMIELLALES (PRASINOPHYCEAE,CHLOROPHYTA)1

A new marine microalga from the Mediterranean Sea, Crustomastix stigmatica Zingone, is investigated by means of LM, SEM, TEM, and pigment and molecular analyses (nuclear‐encoded small subunit [SSU] rDNA and plastid‐encoded rbcL). Pigment and molecular information is also provided for the related species Dolichomastix tenuilepis Throndsen et Zingone. Crustomastix stigmatica has a bean‐shaped cell body 3–5 μm long and 1.5–2.8 μm wide, with two flagella four to five times the body length. The single chloroplast is pale yellow‐green, cup‐shaped, and lacks a pyrenoid. A small bright yellow stigma is located in the mid‐dorsal part of the cell under the chloroplast membrane. An additional accumulation of osmiophilic globules is at times seen in a chloroplast lobe. Cells lack flat scales, whereas three different types of hair‐like scales are present on the flagella. The main pigments of C. stigmatica are those typical of Mamiellales, though siphonein/siphonaxanthin replaces prasinoxanthin and uriolide is absent. The pigment pool of D. tenuilepis is more similar to that of Micromonas pusilla (Butcher) Manton et Parke and of other Mamiellales. The nuclear SSU rDNA phylogeny shows that the inclusion of C. stigmatica and D. tenuilepis in the Mamiellales retains monophyly for the order. The two species form a distinct clade, which is sister to a clade including all the other Mamiellales. Results of rbcL analyses failed to provide phylogenetic information at both the order and species level. No unique morphological or pigment characteristics circumscribe the mamiellalean clade as a whole nor its two daughter clades.     

SEXUALITY AND CYST FORMATION IN THE DINOFLAGELLATE GONYAULAX TAMARENSIS: CYST YIELD IN BATCH CULTURES1

Encystment of the toxic dinoflagellate Gonyaulax tamarensis Lebour (var. excavata) was monitored in batch cultures exposed to a variety of nutritional and environmental treatments. Limitation by nitrogen (as ammonium or nitrate) or phosphorus (as phosphate) resulted in cyst formation. When the initial concentration of limiting nutrient was varied, total cyst yield (mL^?1) was directly proportional to the cell yield at all but the highest nutrient concentrations (where encystment was minimal). Encystment efficiency was relatively constant (0.1–0.2 cysts · cell^?1) over a 5-fold range of cell densities, indicating that 20 to 40% of the vegetative populations successfully encysted. Cyst formation was negligible in nutrient-replete medium, even with a significant reduction in growth rate due to non-optimal light, temperature, or to high batch culture cell densities. Low light levels did decrease cyst yield once encystment was initiated by nutrient limitation, but this was probably linked to smaller motile cell yield and not to a specific inhibition of encystment. In contrast, encystment was more sensitive to temperature than was growth rate: optimal cyst production occurred over a relatively narrow temperature range and no cysts were formed at [Page missing]     

ENCYSTMENT OF THE DINOFLAGELLATE GYRODINIUM UNCATENUM: TEMPERATURE AND NUTRIENT EFFECTS1

Sexual reproduction and encystment of the marine dinoflagellate Gyrodinium uncatenum Hulburt were induced in nitrogen and phosphorus-limited batch cultures. Sexuality did not occur under nutrient-replete conditions even when growth rate was reduced by non-optimal temperatures. Growth was optimal over a broader temperature range than encystment and virtually no cysts were produced at some low and high temperatures where growth occurred. Most cells initiated sexuality as intracellular pools of each limiting nutrient reached minimum or subsistence levels as much as four days after extracellular nutrients were exhausted. High nitrogen cell quotas during the phosphorus experiment indicate that sexuality was induced by a shortage of phosphorus and not by an indirect effect on nitrogen uptake. Total cyst yield corresponded to successful encystment of 9–13% of the motile populations, yet 60–85% of the plateau-phase motile cells were planozygotes (swimming zygotes formed from fusing gametes). Batch culture studies monitoring total cyst yield may thus seriously underestimate the extent of sexuality. More importantly, the number of cysts produced in a dinoflagellate population may be significantly reduced by environmental factors acting on the cells after sexual induction and fusion.     

TWO NEW AUSTRALIAN SPECIES OF PREDAEA (NEMASTOMATACEAE,RHODOPHYTA) WITH TAXONOMIC RECOMMENDATIONS FOR AN EMENDED NEMASTOMATALES AND EXPANDED HALYMENIALES1

Two new species of the red algal genus Predaea are described from Australia. The first, Predaea aurora Kraft et G.W. Saunders, sp. nov., is unusual in its cool‐temperate distribution and in a number of anatomical attributes, most notably the rhizoid‐like habit of the nutritive filaments associated with the auxiliary cells. The second species, Predaea tumescens Kraft et G.W. Saunders, sp. nov., inhabits a coral reef habitat more typical of the genus but nonetheless differs in a number of anatomical details from other reported species. Nuclear small subunit rDNA sequences have confirmed the affinity of P. aurora to other species currently included in this genus. Molecular analyses further indicate that Predaea belongs in the Nemastomataceae and that the Nemastomataceae and Schizymeniaceae are sister taxa in an independent clade of “lineage 4” florideophyte algae. As such, a proposal is made to resurrect the Nemastomatales Kylin emend. G.W. Saunders et Kraft to accommodate these two families. Within the Schizymeniaceae, the Australian‐endemic Platoma australicum and Platoma foliosum were only distantly related to the other included Platoma, Schizymenia, and Titanophora spp. We therefore propose Wetherbeella G.W. Saunders et Kraft, gen. nov., to accommodate these two species. An additional outcome of our molecular analyses is that the genus Tsengia is not a member of the Nemastomataceae (where it is currently placed) but rather forms an independent lineage in the Halymeniales that we now designate as the Tsengiaceae G.W. Saunders et Kraft, fam. nov. Finally, the South African Nemastoma lanceolatum J. Agardh is transferred to Tsengia.     

LIFE CYLE AND SEXUALITY OF THE FRESHWATER RAPHIDOPHYTE GONYOSTOMUM SEMEN (RAPHIDOPHYCEAE)1

Previously unknown aspects in the life cycle of the freshwater flagellate Gonyostomum semen (Ehrenb.) (Raphidophyceae) are described here. This species forms intense blooms in many northern temperate lakes, and has increased in abundance and frequency in northern Europe during the past decades. The proposed life cycle is based on observations of life cycle stages and transitions in cultures. Viable stages of the life cycle were individually isolated and monitored by time‐lapse photography. The most common processes undertaken by the isolated cells were: division, fusion followed by division, asexual cyst formation, and sexual cyst formation. Motile cells divided by two different processes. One lasted between 6 and 24 h and formed two cells with vegetative cell size and with or without the same shape. The second division process lasted between 10 and 20 min and formed two identical cells, half the size of the mother cell. Planozygotes formed by the fusion of hologametes subsequently underwent division into two cells. Asexual cyst‐like stages were spherical, devoid of a thick wall and red spot, and germinated in 24–48 h. Heterogamete pairs were isogamous, and formed an angle of 0–90° between each other. Planozygote and sexual cyst formation were identified within strains established from one vegetative cell. The identity of these strains, which was studied by an amplified fragment length polymorphism analysis, was correlated with the viability of the planozygote. Resting cyst germination was described using cysts collected in the field. The size and morphology of these cysts were comparable with those formed sexually in culture. The excystment rate was higher at 24°C than at 19 or 16°C, although the cell liberated during germination (germling) was only viable at 16°C. The placement of G. semen within the Raphidophyceae family was confirmed by sequence analysis of a segment of the 18S ribosomal DNA.     

EVIDENCE FOR ASEXUAL RESTING CYSTS IN THE LIFE CYCLE OF THE MARINE PERIDINOID DINOFLAGELLATE,SCRIPPSIELLA HANGOEI1

Scrippsiella hangoei (Schiller) Larsen is a peridinoid dinoflagellate that grows during winter and spring in the Baltic Sea. In culture this species formed round, smooth cysts when strains were mixed, indicating heterothallic sexuality and hypnozygote production. However, cysts of the same morphology were also formed in clonal strains exposed to slightly elevated temperature. To better understand the role of cysts in the life cycle of S. hangoei, cyst formation and dormancy were examined in culture experiments and the cellular DNA content of flagellate cells and cysts was compared in clonal and mixed strains using flow cytometry. S. hangoei exhibited a high rate of cyst formation in culture. Cysts produced in both clonal and mixed strain cultures were thick‐walled and underwent a dormancy period of 4 months before germinating. The S. hangoei flagellate cell population DNA distributions consisted of 1C, intermediate, and 2C DNA, indicative of respective eukaryotic cell cycle phases G1, S, and G2M. The majority (>95%) of cysts had a measured DNA content equivalent to the lower 1C DNA value, indicating a haploid nuclear phase and an asexual mode of cyst formation. A small percentage (<5%) of cysts produced in the mixed strain culture had 2C DNA, and thus could have been diploid zygotes. These findings represent the first measurements of dinoflagellate resting cyst DNA content, and provide the first quantitative evidence for dinoflagellate asexual resting cysts. Asexual resting cysts may be a more common feature of dinoflagellate life cycles than previously thought.     

GROWTH AND TOXICITY OF PRYMNESIUM PARVUM (HAPTOPHYTA) AS A FUNCTION OF SALINITY,LIGHT, AND TEMPERATURE1

The growth rate, stationary cell concentration, and toxicity of Prymnesium parvum N. Carter were measured using a strain isolated from Texas inland waters. We used a multifactor experimental approach with multiple regression analysis to determine the importance of environmental factors, including temperature, light, and salinity to these algal measurements. Exponential growth rate was unimodal in relation to temperature, salinity, and irradiance, with an estimated maximal growth of 0.94 d^?1 occurring at 27°C, 22 practical salinity units (psu), and 275 μmol photons·m^?2·s^?1. Stationary cell concentrations also had unimodal responses to temperature and salinity but increased with irradiance. Maximal cell concentrations were estimated to occur at 26°C and 22 psu. Both maximum growth rate and highest stationary cell concentrations were measured at levels of each factor resembling warm, estuarine conditions that differ from the conditions under which blooms occur in inland waters in the southwestern United States. Acute toxicity to fish was highest at the lowest salinity and temperature levels, conditions not optimal for exponential growth but similar to those under which blooms occur in inland waters. Our results imply that summer blooms could occur in inland waters of the southwestern United States. Generally, they have not, suggesting that factors other than those investigated in this research influence bloom dynamics.     

INTERACTIVE EFFECTS OF SALINITY AND TEMPERATURE ON PLANOZYGOTE AND CYST FORMATION OF ALEXANDRIUM MINUTUM (DINOPHYCEAE) IN CULTURE1

The factors regulating dinoflagellate life‐cycle transitions are poorly understood. However, their identification is essential to unravel the causes promoting the outbreaks of harmful algal blooms (HABs) because these blooms are often associated with the formation and germination of sexual cysts. Nevertheless, there is a lack of knowledge on the factors regulating planozygote‐cyst transitions in dinoflagellates due to the difficulties of differentiating planozygotes from vegetative stages. In the present study, two different approaches were used to clarify the relevance of environmental factors on planozygote and cyst formation of the toxic dinoflagellate Alexandrium minutum Halim. First, the effects of changes in initial phosphate (P) and nitrate (N) concentrations in the medium on the percentage of planozygotes formed were examined using flow cytometry. Second, two factorial designs were used to determine how salinity (S), temperature (T), and the density of the initial cell inoculum (I) affect planozygote and resting‐cyst formation. These experiments led to the following conclusions: 1. Low P/N ratios seem to induce gamete expression because the percentage of planozygotes recorded in the absence of added phosphate (‐P) was significantly higher than that obtained in the absence of added nitrogen (‐N), or when the concentrations of both nitrogen and phosphate were 20 times lower (N/20 + P/20). 2. Salinity (S) and temperature (T) strongly affected both planozygote and cyst formation, as sexuality in the population increased significantly as salinity decreased and temperatures increased. S, T combinations that resulted in no significant cyst formation were, however, favorable for vegetative growth, ruling out the possibility of negative effects on cell physiology. 3. The initial cell density is thought to be important for sexual cyst formation by determining the chances of gamete contact. However, the inoculum concentrations tested did not explain either planozygote formation or the appearance of resting cysts.     

THE SEXUAL LIFE CYCLES OF PFIESTERIA PISCICIDA AND CRYPTOPERIDINIOPSOIDS (DINOPHYCEAE)1

Sexual life cycle events in Pfiesteria piscicida and cryptoperidiniopsoid heterotrophic dinoflagellates were determined by following the development of isolated gamete pairs in single‐drop microcultures with cryptophyte prey. Under these conditions, the observed sequence of zygote formation, development, and postzygotic divisions was similar in these dinoflagellates. Fusion of motile gamete pairs each produced a rapidly swimming uninucleate planozygote with two longitudinal flagella. Planozygotes enlarged as they fed repeatedly on cryptophytes. In <12 h in most cases, each planozygote formed a transparent‐walled nonmotile cell (cyst) with a single nucleus. Zygotic cysts did not exhibit dormancy under these conditions. In each taxon, dramatic swirling chromosome movements (nuclear cyclosis) were found in zygote nuclei before division. In P. piscicida, nuclear cyclosis occurred in the zygotic cyst or apparently earlier in the planozygote. In the cryptoperidiniopsoids, nuclear cyclosis occurred inthe zygotic cyst. After nuclear cyclosis, a single cell division occurred in P. piscicida and cryptoperidiniopsoid zygotic cysts, producing two offspring that emerged as biflagellated cells. These two flagellated cells typically swam for hours and fed on cryptophytes before encysting. A single cell division in these cysts produced two biflagellated offspring that also fed before encysting for further reproduction. This sequence of zygote development and postzygotic divisions typically was completed within 24 h and was confirmed in examples from different isolates of each taxon. Some aspects of the P. piscicida sexual life cycle determined here differed from previous reports.     

EFFECT OF SALINITY ON PSEUDO‐NITZSCHIA SPECIES (BACILLARIOPHYCEAE) GROWTH AND DISTRIBUTION1

Salinity varies widely in coastal areas that often have a high abundance of Pseudo‐nitzschia H. Peragallo. Pseudo‐nitzschia is abundant in Louisiana waters, and high cellular domoic acid has been observed in natural samples but no human illness has been reported. To assess the threat of amnesic shellfish poisoning (ASP), we examined the effect of salinity on Pseudo‐nitzschia occurrence in the field and growth in the laboratory with special emphasis on the salinity range where oysters are harvested (10–20 psu). In Louisiana coastal waters, Pseudo‐nitzschia spp. occurred over a salinity range of 1 to >35 psu, but they occurred more frequently at higher rather than lower salinities. Seven species were identified, including toxigenic species occurring at low salinities. In culture studies, seven clones of three species grew over a salinity range of 15 to 40 psu, some grew at salinities down to 6.25 psu, and most grew at salinities up to 45 psu. Tolerance of low salinities decreased from Pseudo‐nitzschia delicatissima (Cleve) Heiden to P. multiseries (Hasle) Hasle to P. pseudodelicatissima (Hasle) Hasle emend. Lundholm, Hasle et Moestrup. In conclusion, although Pseudo‐nitzschia was more prevalent in the field and grew better in the laboratory at higher salinities, it grew and has been observed at low salinities. Therefore, the probability of ASP from consumption of oysters harvested from the low salinity estuaries of the northern Gulf of Mexico is low but not zero; animal mortality events from toxin vectors other than oysters at higher salinity on the shelf are more likely.     

Life history stages of Pyramimonas tychotreta (Prasinophyceae, Chlorophyta), a marine flagellate from the Ross Sea, Antarctica

During a summer cruise to the Ross Sea (Antarctica) areas of snow‐covered sea ice were red‐coloured due to high concentrations of the recently described Pyramimonas tychotreta Daugbjerg. Light microscopy of living material revealed that the population was comprised of quadriflagellate motile cells and thick‐walled cysts. The red colour was due to large numbers of secondary carotenoid‐containing granules, positioned in the periphery of motile cells and cysts. Mature cysts also contained numerous starch grains and lipid droplets. Cells from a red‐coloured field sample turned green overnight as the secondary carotenoids disappeared when cells were placed in low light conditions. The sample then exhibited the typical grass‐green colour of motile cells observed in water samples from the area. Under reduced light motile cells showed strong positive phototaxis. The encystment process involved the asexual transformation of quadriflagellate cells into cysts. A single type of square cyst scale, with perforated floors and walls, replaced the body scales of motile cells. A marked extension, often ending in a hook was at each corner of the cyst scales. Germinating cysts produced four motile cells. Electron microscopy showed the cyst wall to be tri‐layered, with a thin, electron‐dense inner layer, a thick middle layer and a thin outer layer. Sea ice samples with dense populations of motile cells and cyst stages also contained elongate uniflagel‐late cells. These cells were covered with box scales, foot‐print scales, an underlayer of pentagonal scales, limuloid scales and flagellar hair scales identical to those present on the quadriflagellate stage. We tentatively suggest that the uniflagellate stage represents a gamete and its presence implies the occurrence of sexual reproduction. Although, fusion of gametes was not observed, a biflagellate cell with a larger volume was seen which may have been a zygote. How this stage fits into of the life history remains to be explained.     

RESPONSE TO SALINITY AND HEAT STRESS IN TWO HALOTOLERANT CHLOROPHYTE ALGAE1

Two new isolates of halotolerant chlorophyte algae from the Salt Plains National Wildlife Refuge in Oklahoma, USA, tentatively identified as Dunaliella sp. Teodoresco and Nannochloris sp. Naumann, were characterized with respect to interaction between growth salinity and short‐term heat tolerance. Cells were cultured at 23–25° C over a wide range of salinity. In both species, salinity alone had little effect on maximum photochemical yield (measured by pulse modulated fluorescence) and integrity of the light harvesting system (77 K fluorescence emission spectra). In contrast, Nannochloris exhibited decreasing growth rate (μ), light‐saturated photosynthetic capacity (P^cell_max), respiration (R_d), light‐harvesting efficiency (α^cell), and chl content with increasing salinity. Cultures were heated for 2 h near their upper temperature limits (41.5° C for Dunaliella and 45° C for Nannochloris grown at 50 psu). Dunaliella was progressively more heat‐tolerant with increasing salinity. Photochemical yield of cells at 100 and 50 psu was inhibited by about 15% and 40%, respectively, and largely recovered within 30 min after return to 23° C. Thermal inhibition of photochemical yield in Nannochloris was about 45% at both 50 and 100 psu, but recovery was slower at 100 psu. At 20 psu, both species were almost 90% inhibited by high temperature and required more than a day to recover. In both species, 2 h of heating increased the PSI:PSII fluorescence emission ratio (714:690 nm) at all salinities. This ratio largely recovered within 24 h in Dunaliella at 50 and 100 psu and partially recovered in Nannochloris at 100 psu, but cells of both species heated at 20 psu were chlorotic the next day.     

POTENTIAL IMPORTANCE OF BENTHIC CYSTS OF GONYAULAX TAMARENSIS AND G. EXCAVATA IN INITIATING TOXIC DINOFLAGELLATE BLOOMS1,2,3

Thick-walled, nonmotile cysts (termed hypnocysts) of two dinoflagellates were isolated from estuarine sediments in Cape Cod, Massachusetts, and germinated to produce their respective motile, thecate stages. Hypnocysts from Orleans district were identified as Gonyaulax excuvata (Braarud) Balech sensu Loeblich & Loeblich. Visually identical hypnocysts from Falmouth district were provisionally identified as Gonyaulax tamarensis Lebour. Both species were toxic. A geographic survey in September detected hypnocysts in only the sediments of locations where toxic blooms developed the preceding and following Spring. Laboratory incubation (16 C) of hypnocysts from sediment samples stored in the dark (5 C) for 6 mo initiated excystment by the temperature increase, with no appreciable effect from light regime, nutrient, or chelator concentrations. Motility of excysted germlings was optimum in highly chelated medium and in the presence of light. We conclude that hypnocysts of both tasa are important in seeding recurrent annual blooms, synchronizing early bloom development with vernal warning of seawater and increasing the geographic range of the species. We suggest that many red tides in New England and eastern Canadian waters are initiated through the displacement of motile estuarine populations into nearshore area by tidal advection and surface runoff, although the potential existence and importance of offshore cyst reservoirs cannot be discounted. Evidence is presented that hypnocysts are probable sexual zygotes whereas the thin-walled cysts readily formed in laboratory cultures (pellicle cyst) are asexual. Pellicle cysts are of limited durability, do not overwinter in nature, and therefore do not play a significant role in initiating toxic blooms.     

Process length variation in cysts of a dinoflagellate,Lingulodinium machaerophorum,in surface sediments: Investigating its potential as salinity proxy

A biometrical analysis of the dinoflagellate cyst Lingulodinium machaerophorum [Deflandre, G., Cookson, I.C., 1955. Fossil microplankton from Australia late Mesozoic and Tertiary sediments. Australian journal of Marine and Freshwater Research 6: 242–313.] Wall, 1967 in 144 globally distributed surface sediment samples revealed that the average process length is related to summer salinity and temperature at a water depth of 30 m by the equation (salinity/temperature) = (0.078?average process length + 0.534) with R² = 0.69. This relationship can be used to reconstruct palaeosalinities, albeit with caution. The particular ecological window can be associated with known distributions of the corresponding motile stage Lingulodinium polyedrum (Stein) Dodge, 1989. Confocal laser microscopy showed that the average process length is positively related to the average distance between process bases (R² = 0.78), and negatively related to the number of processes (R² = 0.65). These results document the existence of two end members in cyst formation: one with many short, densely distributed processes and one with a few, long, widely spaced processes, which can be respectively related to low and high salinity/temperature ratios. Obstruction during formation of the cysts causes anomalous distributions of the processes. From a biological perspective, processes function to facilitate sinking of the cysts through clustering.     

THE CALCAREOUS RESTING CYST OF PENTAPHARSODINIUM TYRRHENICUM COMB. NOV. (DINOPHYCEAE)1

While investigating dinoflagellate cyst assemblages in surface sediments of the Gulfs of Naples and Salerno (Mediterranean Sea), we found a new calcareous resting cyst. This cyst has a round to oval body surrounded by a thick mineral layer, which gives it the shape of a Napoleon hat, with a flat, oval face bearing the archeopyle and a convex keel on the opposite side. The cyst shape is variable in both natural samples and clonal cultures. The organic membrane underlying the calcareous covering is resistant to acetolysis, thus demonstrating the presence of sporopolleninlike material. The cyst germinated into a motile stage having the same morphological features and thecal plate pattern as Peridinium tyrrhenicum Balech. We believe the validity of the genus Pentapharsodinium Indelicato & Loeblich should be accepted. Based on the comparative examination of the species we collected and of a similar species, Pentapharsodinium trachodium Indelicato & Loeblich, we propose Pentapharsodinium tyrrhenicum as a new combination for Peridinium tyrrhenicum. The genus Pentapharsodinium also includes P. dalei Indelicato & Loeblich (= Peridinium faeroense Dale), which produces spiny, organic-walled cysts. The presence of species forming calcareous cysts and species producing noncalcareous cysts in the same genus raises questions about maintaining the family Calciodinellaceae. This family should only include calcareous cyst-forming peridinioids, in order to maintain a unified system of classification of fossil and recent dinoflagellates.     

ROLE OF ENCYSTMENT AND EXCYSTMENT OF PERIDINIUM BIPES F. OCCULATUM (DINOPHYCEAE) IN FRESH WATER RED TIDES IN LAKE KIZAKI,JAPAN1

The encystment flux of Peridinium bipes f. occulatum (Dinophyceae) was investigated with sediment traps from 1968 to 1990 in Lake Kazki. Cysts of P. bipes were formed throughout the blooms, Encystment flux of P. bipes in the pelagic zone was usually lower than those at shallow sites, and the density of P. bipes cysts in lake sediment was higher in the shallow region than in the pelagic zone. However, in the shallower region, The concentration of P. bipes cysts varied widely, possibly due to high rates of encystment and excystment. Peridinium bipes encystment occurred between 15° and 25° C in the laboratory, with very little cyst formation below 10°C. Though cyst formation was observed in continous darkness, the rate increased with irradiance. Under continuous darkness, no excystment was observed at any temperature from 5° to 25° C. Eighty-one percent of the cysts illuminated at 105 μE m^?2 s^?1 excysted after 13 days incubation at 15° C, and lower irradiances decreased germination success. Results from laboratory experiments suggest that light is a critical factor in the germination of P. bipes cysts. Bottom depth thus can have a significant effect on germination because cysts only can excyst from depths where light is sufficient. The shallow region of the lake is thus very important as a seed bed for P. bipes during early spring. Cyst deposited in deeper waters may not ever germinate unless they are resuspended and transported to shallow areas where light reaches the bottom.     

The effect of salinity on the growth,toxin production,and morphology of <Emphasis Type="BoldItalic">Nodularia spumigena</Emphasis> isolated from the Gulf of Gdańsk,southern Baltic Sea

Culture experiments on the toxic Nodularia spumigena strain NSGG-1 isolated from the Gulf of Gdańsk showed a significant effect of salinity on growth and nodularin production. Growth of the NSGG-1 strain, was optimal between 7 and 18 psu, lower at 3 and 24 psu and was significantly inhibited at the extreme salinities of 0 and 35 psu. Nodularin (NOD) content of N. spumigena, estimated by the NOD/Chla ratios, correlated positively with salinity and increased from 0 to 35 psu. The NOD/Chla ratio on day 10 of growth was high, and, reached the maximum at day 30. A sudden increase in salinity from 7 to 18 and 35 psu resulted in plasmolysis of Nodularia cells. Salinity was also observed to have other effects on NSGG-1; the filaments were longest at 7 psu, while an increased number of akinetes were formed at 35 psu. The number of heterocytes was markedly reduced at the extreme salinities. This latter finding might explain why Nodularia blooms do not occur outside a certain salinity range in nitrogen-deficient waters.