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1.
Alexandrium peruvianum (Balech et Mendiola) is a noxious phototrophic marine dinoflagellate. During the life cycle of this species, two kinds of cysts are produced: resting cysts, which are long-lasting and double-walled, and temporary cysts, which are short-lasting and thin-walled. In addition, short-lasting, but resting-like cysts can also be formed. Although it is crucial to identify sexual events in a dinoflagellate population, sexual and asexual cysts are morphologically very similar in this species. Therefore, we studied the complete life cycle and the nature of the cyst-like stages formed after individual isolation of specimens and crossing of clonal cultures established from germination of wild resting cysts. Asexual division in A. peruvianum takes place either in the motile stage by sharing of the theca (desmoschisis), or inside a vegetative cyst (temporary cyst), from which two, or at times four or six naked daughter cells can originate. The daughter cells completely synthesize new cell walls (eleutheroschisis). Sexuality was confirmed by the presence of fusing gamete pairs and longitudinally biflagellated planozygotes after out-crossing of compatible clonal strains. However, the clonal cultures had low levels of self-compatibility, since a flow cytometry analysis showed that synchronized self-crosses produced few zygotes (<5%). After isolation of individual cells, it was proved that the fate of the planozygotes depended on the nutritional status of the isolation media. Most of the planozygotes isolated to replete medium (L1) divided, whereas in medium lacking nitrates (L-N) or phosphates (L-P) they formed temporary, thin-walled cysts. Temporary cysts formed in L1 were always uninucleated and gave rise to one cell, while those formed in L-N or L-P produced 1–6 small cells. In addition, resting cysts were formed in culture, but never after individual planozygote isolation. Resting cysts were uninucleated and needed maturation time before entering dormancy. The resting cysts were considered sexual products, since longitudinally biflagellate germlings were liberated after germination in all cases studied. Mature resting cysts (52.3 ± 3.0 μm) had a dormancy period of 1–3 months, whereas temporary asexual cysts (32.5 ± 5.4 μm) germinated in less than 7 days.  相似文献   

2.
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.  相似文献   

3.
Between May 2000 and February 2001, a major bloom of the toxic dinoflagellate Gymnodinium catenatum (a causative organism of Paralytic Shellfish Poisoning, PSP) affected over 1500 km of coastline of New Zealand’s North Island. As this was the first record of this species in New Zealand, we aimed to resolve whether this represented a recent introduction/spreading event or perhaps an indigenous cryptic species stimulated by environmental/climatic change. To answer this question, we analysed for G. catenatum resting cysts in 210Pb dated sediment cores (18–34 cm long; sedimentation rates 0.34–0.69 cm per year) collected by SCUBA divers from Manukau Harbour, where the species was first detected, and from Hokianga Harbour, where the highest shellfish toxicity was recorded, while using Wellington Harbour as a well-monitored control site. The results of this study conclusively demonstrate that abundant G. catenatum has been in northern New Zealand at least since the early 1980s, increasing up to 1200 cysts/g around the year 2000, but with low cyst concentrations possibly present since at least 1937. In contrast, Wellington Harbour cores contained only very sparse G. catenatum cysts (8 cysts/g), present only to a depth of 7 cm (surface mixed layer depth), reflecting an apparent recent range expansion of this dinoflagellate in New Zealand, possibly stimulated by unusual climatic conditions associated with the 2000 La Nina event. The significant increases since the early 1980s also of Protoperidinium cysts at Hokianga Harbour and of Gonyaulax, Protoperidinium and Protoceratium cysts at Manukau Harbour suggest a broad scale environmental change has occurred in Northland, New Zealand.  相似文献   

4.
One proposed solution to the problem of ballast-mediated aquatic invasions involves chemically treating ballast water to kill key target organisms. Here, we examine the efficacy of three commercially available ballast water biocides using vegetative microalgae, dinoflagellate resting cysts and bacteria as test organisms. Chemicals tested were the ballast water biocides SeaKleen® and Peraclean® Ocean, and the chlorine dioxide biocide Vibrex®. Results demonstrate that the applicability of each of the three chemical biocides as a routine ballast water treatment is limited by factors such as cost, biological effectiveness and possible residual toxicity of the discharged ballast water (assessed on the basis of impact on motility of vegetative marine microalgae). Of the three biocides tested, Peraclean® Ocean appears to hold the most potential; however its effectiveness in shipboard trials is yet to be proven. Peraclean® Ocean was biodegradable within 2–6 weeks (initial concentration of 200 ppm), could effectively inactivate resting cysts of the marine dinoflagellates Gymnodinium catenatum, Alexandrium catenella and Protoceratium reticulatum at 400 ppm, could control bacterial growth of Escherichia coli, Staphylococcus aureus, Listeria innocua and Vibrio alginolyticus at 125–250 ppm, and could eliminate vegetative dinoflagellate cells at a concentration of 100 ppm. SeaKleen® eliminated vegetative microalgae at 2 ppm and could control resting cysts of the dinoflagellates G. catenatum and P. reticulatum at a concentration of 6 and 10 ppm, respectively, when exposed for a period of 2 weeks. SeaKleen® did not inactivate resting cysts of A. catenella at a concentration of 10 ppm and was found to degrade at a rate that could result in the discharge of residual toxic water into the marine environment. Together with the poor bactericidal properties of SeaKleen® (100–200 ppm required), this may limit the use of this biocide as a routine treatment option. Vibrex® is not a suitable ballast water treatment option due to the need for hydrochloric acid as an activator, however it was found to be the most effective against bacteria (complete inhibition at 15 ppm) indicating that onboard chlorine dioxide generators may provide an effective bacterial treatment option. The performance of these biocides was adversely influenced by a variety of factors including low water temperatures (6 °C compared to 17 °C), light versus dark conditions, and the presence of humus-rich seawater and ballast water sediments.  相似文献   

5.
Interactions with the bacterial community are increasingly considered to have a significant influence on marine phytoplankton populations. Here we used a simplified dinoflagellate‐bacterium experimental culture model to conclusively demonstrate that the toxic dinoflagellate Gymnodinium catenatum H. W. Graham requires growth‐stimulatory marine bacteria for postgermination survival and growth, from the point of resting cyst germination through to vegetative growth at bloom concentrations (103 cells · mL?1). Cysts of G. catenatum were germinated and grown in unibacterial coculture with antibiotic‐resistant or antibiotic‐sensitive Marinobacter sp. DG879 or Brachybacterium sp., and with mixtures of these two bacteria. Addition of antibiotics to cultures grown with antibiotic‐sensitive strains of bacteria resulted in death of the dinoflagellate culture, whereas cultures grown with antibiotic‐resistant bacteria survived antibiotic addition and continued to grow beyond the 21 d experiment. Removal of either bacterial type from mixed‐bacterial dinoflagellate cultures (using an antibiotic) resulted in cessation of dinoflagellate growth until bacterial concentration recovered to preaddition concentrations, suggesting that the bacterial growth factors are used for dinoflagellate growth or are labile. Examination of published reports of axenic dinoflagellate culture indicate that a requirement for bacteria is not universal among dinoflagellates, but rather that species may vary in their relative reliance on, and relationship with, the bacterial community. The experimental model approach described here solves a number of inherent and logical problems plaguing studies of algal‐bacterium interactions and provides a flexible and tractable tool that can be extended to examine bacterial interactions with other phytoplankton species.  相似文献   

6.
Cryothecomonas longipes Schnepf and Kühn sp. nov. is a colourless biflagellate organism, 9–14 μm long and 7–9 μm wide when not filled with food vacuoles. It was detected in the North Sea, feeding with pseudopodia on diatoms. It penetrates the host shell, while the main body of the flagellate remains outside the frustule. Cells are covered with a multilayered theca. The pseudopodium protrudes through a preformed slit in the theca. Each flagellum also emerges through a pit in which the theca forms a funnel of complex structure that girdles each flagellum. The anterior flagellum is 9–15 μm long and oriented forward; the ventral flagellum, posteriorly directed, is 20–24 μm long and bears fine hairs. The flagellar roots consist of microtubules that emerge at satellites around the basal bodies and run along the flagellar pits. In addition, the ventral flagellum is accompanied by a band of six microtubules. It is proximally attached to a small fibrillar band, which interconnects the basal bodies. Cryothecomonas longipes has two or three types of extrusomes which pierce the theca when discharged. Their mode of discharge is discussed. Microbody-like vesicles containing small tubules are closely associated with older digestion vacuoles. Cryothecomonas longipes is compared with other species of the genus and a diagnosis is given. Received: 4 March 1999 / Received in revised form: 28 July 1999 / Accepted: 30 August 1999  相似文献   

7.
8.
A small, freshwater dinoflagellate with an incomplete cingulum, identified as Esoptrodinium gemma Javornický (=Bernardinium bernardinense sensu auctt. non sensu Chodat), was maintained in mixed culture and examined using light and serial section TEM. Vegetative flagellate cells, large cells with two longitudinal flagella (planozygotes), and cysts were examined. The cells displayed a red eyespot near the base of the longitudinal flagellum, made of two or three layers of pigment globules not bounded by a membrane. Yellow‐green, band‐shaped chloroplasts, bounded by three membranes and containing lamella with three thylakoids, were present in both flagellate cells and cysts. Most cells had food vacuoles, containing phagotrophically ingested chlamydomonads or chlorelloid green algae; ingestion occurred through the ventral area, involving a thin pseudopod apparently driven by the peduncle. The pusule was tubular, with numerous diverticula in its distal portion, and opened into the longitudinal flagellar canal. Three roots were associated with each pair of flagellar bases, both in vegetative cells and in a planozygote. The longitudinal microtubular root bifurcated around the longitudinal basal body. The planozygote contained a single peduncle and associated structures, and a single transverse flagellar canal with the two converging transverse flagella. Using two ciliates as outgroup species, phylogenetic analyses based on maximum parsimony, neighbor‐joining and posterior probability (Bayesian analysis) supported a clade comprising Esoptrodinium, Tovellia, and Jadwigia.  相似文献   

9.
Data on the qualitative and quantitative composition of resting stages of planktonic microalgae in recent marine sediments of Peter the Great Bay (Sea of Japan) over the period 2000–2007 are presented. A total of sixty one morphological forms of resting stages represented by dinoflagellate and raphidophyte cysts and diatom spores and resting cells were recorded in the sediment samples. This study revealed cysts of the potentially toxic species Alexandrium tamarense, A. cf. minutum, Alexandrium sp., Gymnodinium catenatum (PSP toxin producers), and Protoceratium reticulatum (yessotoxin producer); resting cells of Pseudo-nitzschia sp. (potential producer of domoic acid); and cysts of bloom-forming species Cochlodinium cf. polykrikoides and Heterosigma cf. akashiwo.  相似文献   

10.
11.
We describe a new organic-walled resting cyst from surface sediments of Imari Bay in western Japan. The cysts are spherical, 23–29 pm in diameter, and their surface is covered with spinous to membranous ornaments that are 5–7 μm long and 1.5–2.2 μm wide. The ornaments vary from slender and bifurcate to membranous and multifurcate distal extremities. No archeopyle was observed. The cyst shape is variable in both natural samples and clonal cultures. Vegetative cells are small and ovoid, 17–25 μm long and 14–21 μm wide, and are yellow-brown in color. The epitheca is conical with a conspicuous apical horn, and the hypotheca is hemispherical. The cingular transitional plate has a needle-like spine at its anterior right corner. The plate formula is Po, X, 4″3a, 7″, 5c, 5s 5″and 2″. Although vegetative cells of the present species correspond to Ensiculifera, it is distinct from other species in producing no calcareous cysts. No species of Ensiculifera has been reported to produce cysts composed of only an organic wall. The present species is provisionally placed in the genus Ensiculifera as E. imariense sp. nov.  相似文献   

12.
Cysts of the freshwater dinoflagellate Peridinium are typically different from vegetative cells in shape and remain largely undescribed. Molecular discrimination of such cysts would be useful to a number of research disciplines. A reliable method for the amplification of ribosomal DNA (rDNA) from Peridinium at different life stages is described. This genotyping strategy relies on whole cell PCR using Peridinium-specific primers designed from available 18S rDNA sequences. Here, we demonstrate the effectiveness of Peridinium-specific PCR for the rapid molecular identification of Peridinium cells in various life stages such as vegetative, planozygote, hypnozygote and cyst.  相似文献   

13.
Gymnodinium catenatum Graham (Dinophyceae)in Europe: a growing problem?   总被引:1,自引:0,他引:1  
The microreticulate resting cyst of the potentially toxic, chain-forming,unarmoured neritic dinoflagellate Gymnodinium catenalum Graham1943. the planktonic stage of which is not known from NorthEuropean waters, is reported for the first time from recentGerman coastal sediments of the North Sea and Baltic Sea. Insandy mud sediments of the German Bight, a maximum of 8 5 livingcysts cm–3 were found. In Kiel Bight sediments G.catenalumwas found in maximum concentrations of 17.0 living cysts cm–3.In surface waters of the German Bight resuspended G catenatumcysts were observed at concentrations of up to 3.6 cysts l–1.Successful germination experiments conducted with natural seawatershow that the occurrence of a vegetative form of G.catenatumin northern Europe is very likely. The present study highlightsthat cyst surveys provide an important tool for the evaluationof areas with potential toxicity problems, as they may indicatethe presence of hitherto overlooked species in the water column.  相似文献   

14.
Summary Thalassomyces marsupii, an ellobiopsid parasite of the pelagic amphipod Parathemisto pacifica, produces biflagellate swarmers following simultaneous cleavage of the gonomere. One flagellum is directed posteriorly, the other circumferentially. The vegetative thallus is bounded by a complex pellicle and not a cell wall. Our observations suggest that Thalassomyces is a member of the achlorophyllous Dinophyceae.  相似文献   

15.
Primuline staining is widely used to visualize and enumerate dinoflagellate cysts in marine sediments. In staining cysts of Gymnodinium catenatum H. W. Graham, Scrippsiella trochoidea (F. Stein) A. R. Loebl., and cysts from estuarine sediments, we found their green fluorescence after primuline treatment to be seemingly no different from the green autofluorescence (GAF) inherent in vegetative cells and cysts of dinoflagellates fixed in formaldehyde. Although primuline subsequently proved to enhance green fluorescence of both species quantitatively, we nonetheless recommend taking advantage of dinoflagellates' GAF to detect and count their cysts in sediments. Doing so will reduce the time, chemical consumption, and possible loss of cells involved in the primuline‐staining procedure.  相似文献   

16.
Asexual and sexual life cycle events were studied in cultures of the toxic marine dinoflagellate Protoceratium reticulatum. Asexual division by desmoschisis was characterized morphologically and changes in DNA content were analyzed by flow cytometry. The results indicated that haploid cells with a C DNA content occurred only during the light period whereas a shift from a C to a 2C DNA content (indicative of S phase) took place only during darkness. The sexual life cycle was documented by examining the mating type as well as the morphology of the sexual stages and nuclei. Gamete fusion resulted in a planozygote with two longitudinal flagella, but longitudinally biflagellated cells arising from planozygote division were also observed, so one of the daughter cells retained two longitudinal flagella while the other daughter cell lacked them. Presumed planozygotes (identified by their longitudinally biflagellated form) followed two life-cycle routes: division and encystment (resting cyst formation). Both the division of longitudinally biflagellated cells and resting cyst formation are morphologically described herein. Resting cyst formation through sexual reproduction was observed in 6.1% of crosses and followed a complex heterothallic pattern. Clonal strains underwent sexuality (homothallism for planozygote formation and division) but without the production of resting cysts. Ornamental processes of resting cysts formed from the cyst wall under an outer balloon-shaped membrane and were fully developed in <1 h. Obligatory dormancy period was of ∼4 months. Excystment resulted in a large, rounded, pigmented, longitudinally biflagellated but motionless, thecate germling that divided by desmoschisis. Like the planozygote, the first division of the germling yielded one longitudinally biflagellated daughter cell and another without longitudinal flagella. The longitudinal biflagellation state of both sexual stages and of the first division products of these cells is discussed.  相似文献   

17.
The toxic dinoflagellate Karenia mikimotoi has been well-known for causing large-scale and dense harmful algal blooms (HABs) in coastal waters worldwide and serious economic loss in aquaculture and fisheries and other adverse effects on marine ecosystems. Whether K. mikimotoi forms resting cysts has been a puzzling issue regarding to the mechanisms of bloom initiation and geographic expansion of this species. We provide morphological and molecular confirmation of sexually produced thin-walled resting cysts by K. mikimotoi based on observations of laboratory cultures and their direct detection in marine sediments. Light and scanning electron microscopy evidences for sexual reproduction include attraction and pairing of gametes, gamete fusion, formation of planozygote and thin-walled cyst, and the documentation of the thin-walled cyst germination processes. Evidence for cysts in marine sediments was in three aspects: positive PCR detection of cysts using species-specific primers in the DNA extracted from whole sediments; fluorescence in situ hybridization detection of cysts using FISH probes; and single-cell PCR sequencing for cysts positively labeled with FISH probes. The existence of sexually produced, thin-walled resting cysts by K. mikimotoi provides a possible mechanism accounting for the initiation of annually recurring blooms at certain regions and global expansion of the species during the past decades.  相似文献   

18.
Studies considering the biology and ecology of the toxic bloom‐forming species, Alexandrium pseudogonyaulax, are rare. Our results highlight five features not described before in A. pseudogonyaulax life cycle: (i) A. pseudogonyaulax gametes showed two modes of conjugation, anisogamy and isogamy, (ii) sexual conjugation occurs either in the dark or in the light phase by engulfment or a fusion process, (iii) the presence of planozygote and newly formed cysts in monoclonal culture suggests homothallism, (iv) newly formed cysts have very dark vesicular content and are mostly unparatabulated when observed under light microscope and (v) natural resting cysts are able to give either a planomeiocyte or two vegetative cells. Cyst viability was enhanced after 5 months of cold storage (4°C), with excystment rate reaching 97% after 3 d of incubation. Excystment rate was highest (43%–79%) in Enriched Natural Sea Water diluted culture medium, whereas few germling cells were able to survive without the culture medium (0%–13%). Salinity‐irradiance experiments revealed that the highest cell concentrations occur at high irradiances for all the tested salinities. Vegetative growth rates generally increased with increasing irradiance, and were less dependent on salinity variations. The relatively low growth rate, low cell densities in the laboratory, and the notable capacity of producing cysts along growth phases of A. pseudogonyaulax could explain the occurrence of high resting cysts densities in the sediment of Bizerte lagoon and the relatively low abundances of vegetative cells in the water column.  相似文献   

19.
The composition and metabolic activity of cysts of the marine dinoflagellate Scrippsiella trochoidea (Stein) Loeblich were examined during dormancy, quiescence, and germination. On a per cell basis, newly formed cysts contained an order of magnitude more carbohydrate but significantly less protein and chlorophyll a than did exponentially growing vegetative cells. Loss of lipid and carbohydrate from cysts during the initial dormancy period reflected a respiration rate estimated to be 10% of the respiratory activity in vegetative cells. Among older, quiescent cysts the calculated respiration rate decreased further to approximately 1.5% of the vegetative rate and appeared to proceed largely at the expense of carbohydrate reserves. These estimated rates of respiration were in good agreement with direct measurements of cyst oxygen consumption. The transfer of quiescent cysts to conditions permissive for germination resulted in a rapid increase in respiration rate, as evidenced by carbohydrate loss and O2 consumption. The increased respiratory activity was followed by an increase in protein content and, later, by an increase in chlorophyll a content and photosynthetic capacity. Just prior to germination the P/R ratio became greater than 1, and the estimated chlorophyll-specific photosynthetic activity reached 75% of the rate in vegetative cells. Complete restoration of photosynthetic and respiratory capacity apparently was not achieved until after excystment. These data confirm the common assumption that dinoflagellate cysts represent true “resting” cells, containing extensive energy reserves and displaying greatly reduced metabolic activity.  相似文献   

20.
In this study, we examined the impact of environmental perturbation on the movement of the toxic bloom‐forming alga Heterosigma akashiwo (Hada) Hada ex Y. Hara et Chihara [syn. H. carterae (Hulburt) F.J.R. Taylor] between vegetative and resting cell phases of the life history. Resting state induction, in batch culture, was most effective when vegetative cells were subjected to low temperature (10° C) and darkness for extended time periods. Heterosigma cells in stasis had neither a cell wall nor scales but were surrounded by a calyx, most probably of polysaccharide composition. The resting cell was completely immobile, although both flagella remained attached. Heterosigma resting cells did not require a maturation period before successful activation to the vegetative state could occur. Cell division and motility were impacted sequentially during both the induction and activation phases of resting cell development. Our data show that Heterosigma had an obligate light requirement for resting cell activation. In replete medium, very low light fluences of 5 μmol photons·m ? 2·s ? 1 were as effective as 60 μmol photons·m ? 2·s ? 1 in the initiation of activation. Such sensitivity to extremely low light might give Heterosigma a competitive advantage for bloom formation in nature. Reduced nitrate levels significantly shortened the temporal transition of vegetative cells into the resting cell phase of the life history. Additionally, when resting cells induced in nitrate‐limited medium were activated under nitrate‐replete condition, the efficiency of the activation response was directly correlated to light availability. Both vegetative and resting cells maintained a haploid DNA complement. Rapid amplified polymorphic DNA (RAPD) analysis demonstrated variation in genetic identity among axenic Heterosigma strains. Strain identity influenced success in resting cell induction and survival in stasis. To date, no defined sexual cycle has been described. These observations are discussed in terms of population fitness. The data presented in this report provide a model algal system wherein the molecular events that govern long‐term stasis in an obligately autotrophic organism can now be assessed.  相似文献   

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