Alexandrium ostenfeldii is present in a wide variety of environments in coastal areas worldwide a... more Alexandrium ostenfeldii is present in a wide variety of environments in coastal areas worldwide and is the only dinoflagellate known species that produces paralytic shellfish poisoning (PSP) toxins and two types of cyclic imines, spirolides (SPXs) and gymnodimines (GYMs). The increasing frequency of A. ostenfeldii blooms in the Baltic Sea has been attributed to the warming water in this region. To learn more about the optimal environmental conditions favoring the proliferation of A. ostenfeldii and its complex toxicity, the effects of temperature and salinity on the kinetics of both the growth and the net toxin production of this species were examined using a factorial design and a response-surface analysis (RSA). The results showed that the growth of Baltic A. ostenfeldii occurs over a wide range of temperatures and salinities (12.5–25.5°C and 5–21, respectively), with optimal growth conditions achieved at a temperature of 25.5°C and a salinity of 11.2. Together with the finding that a salinity > 21 was the only growth-limiting factor detected for this strain, this study provides important insights into the autecology and population distribution of this species in the Baltic Sea. The presence of PSP toxins, including gonyautoxin (GTX)-3, GTX-2, and saxitoxin (STX), and GYMs (GYM-A and GYM-B/-C analogues) was detected under all temperature and salinity conditions tested and in the majority of the cases was concomitant with both the exponential growth and stationary phases of the dinoflagellate’s growth cycle. Toxin concentrations were maximal at temperatures and salinities of 20.9°C and 17 for the GYM-A analogue and > 19°C and 15 for PSP toxins, respectively. The ecological implications of the optimal conditions for growth and toxin production of A. ostenfeldii in the Baltic Sea are discussed.
Dinoflagellates are haploid eukaryotic microalgae in which rapid proliferation causes dense bloom... more Dinoflagellates are haploid eukaryotic microalgae in which rapid proliferation causes dense blooms, with harmful health and economic effects to humans. The proliferation mode is mainly asexual, as the sexual cycle is believed to be rare and restricted to stressful environmental conditions. However, sexuality is key to explaining the recurrence of many dinoflagellate blooms because in many species the fate of the planktonic zygotes (planozygotes) is the formation of resistant cysts in the seabed (encystment). Nevertheless, recent research has shown that individually isolated planozygotes in the lab can enter other routes besides encystment, a behavior of which the relevance has not been explored at the population level. In this study, using imaging flow cytometry, cell sorting, and Fluorescence In Situ Hybridization (FISH), we followed DNA content and nuclear changes in a population of the toxic dinoflagellate Alexandrium minutum that was induced to encystment. Our results first show that planozygotes behave like a population with an “encystment-independent” division cycle, which is light-controlled and follows the same Light:Dark (L:D) pattern as the cycle governing the haploid mitosis. Resting cyst formation was the fate of just a small fraction of the planozygotes formed and was restricted to a period of strongly limited nutrient conditions. The diploid-haploid turnover between L:D cycles was consistent with two-step meiosis. However, the diel and morphological division pattern of the planozygote division also suggests mitosis, which would imply that this species is not haplontic, as previously considered, but biphasic, because individuals could undergo mitotic divisions in both the sexual (diploid) and the asexual (haploid) phases. We also report incomplete genome duplication processes. Our work calls for a reconsideration of the dogma of rare sex in dinoflagellates.
Parasites exert important top-down control of their host populations. The host-parasite system fo... more Parasites exert important top-down control of their host populations. The host-parasite system formed by Alexandrium minutum (Dinophyceae) and Parvilucifera sinerae (Perkinsozoa) offers an opportunity to advance our knowledge of parasitism in planktonic communities. In this study, DNA extracted from 73 clonal strains of P. sinerae, from 10 different locations along the Atlantic and Mediterranean coasts, was used to genetically characterize this parasitoid at the species level. All strains showed identical sequences of the small and large subunits and internal transcribed spacer of the ribosomal RNA, as well as of the β-tubulin genes. However, the phenotypical characterization showed variability in terms of host invasion, zoospore success, maturation time, half-maximal infection, and infection rate. This characterization grouped the strains within 3 phenotypic types distinguished by virulence traits. A particular virulence pattern could not be ascribed to host-cell bloom appearance or to the location or year of parasite-strain isolation; rather, some parasitoid strains from the same bloom significantly differed in their virulence traits. Identical markers such as ITS and β-tubulin genes of P. sinerae strains from different geographic areas and from different years precludes their use in assessing intra-specific diversity and could indicate a recent dispersion of this species.
Abstract
Background: Marine microbial protists, in particular, dinoflagellates, produce polyketi... more Abstract
Background: Marine microbial protists, in particular, dinoflagellates, produce polyketide toxins with ecosystem-wide and human health impacts. Species of Gambierdiscus produce the polyether ladder compounds ciguatoxins and maitotoxins,
which can lead to ciguatera fish poisoning, a serious human illness associated with reef fish consumption. Genes
associated with the biosynthesis of polyether ladder compounds are yet to be elucidated, however, stable isotope feeding studies of such compounds consistently support their polyketide origin indicating that polyketide synthases are
involved in their biosynthesis.
Results: Here, we report the toxicity, genome size, gene content and transcriptome of Gambierdiscus australes and G.
belizeanus. G. australes produced maitotoxin-1 and maitotoxin-3, while G. belizeanus produced maitotoxin-3, for which cell extracts were toxic to mice by IP injection (LD50 = 3.8 mg kg-1). The gene catalogues comprised 83,353 and 84,870
unique contigs, with genome sizes of 32.5 ± 3.7 Gbp and 35 ± 0.88 Gbp, respectively, and are amongst the most
comprehensive yet reported from a dinoflagellate. We found three hundred and six genes involved in polyketide
biosynthesis, including one hundred and ninty-two ketoacyl synthase transcripts, which formed five unique
phylogenetic clusters.
Conclusions: Two clusters were unique to these maitotoxin-producing dinoflagellate species, suggesting that they may
be associated with maitotoxin biosynthesis. This work represents a significant step forward in our understanding of the genetic basis of polyketide production in dinoflagellates, in particular, species responsible for ciguatera fish poisoning.
Dinoflagellates are microscopic aquatic eukaryotes with huge genomes and an unusual cell regulati... more Dinoflagellates are microscopic aquatic eukaryotes with huge genomes and an unusual cell regulation. For example, most genes are present in numerous copies and all copies seem to be obligatorily transcribed. The consequence of the gene copy number for final protein synthesis is however not clear. One such gene is sxtA, the starting gene of paralytic shellfish toxin (PST) synthesis. PSTs are small neurotoxic compounds that can accumulate in the food chain and cause serious poisoning incidences when ingested. They are produced by dinoflagellates of the genera Alexandrium, Gymnodium and Pyrodinium.
Here we investigated if the genomic copy number of sxtA4 is related to PST content in Alexandrium minutum cells. SxtA4 is the 4th domain of the sxtA gene and its presence is essential for PST synthesis in dinoflagellates. We used PST and genome size measurements as well as quantitative PCR to analyze sxtA4 copy number and toxin content in 15 A. minutum strains. Our results show a strong positive correlation between the sxtA4 copy number and the total amount of PST produced in actively growing A. minutum cells. This correlation was independent of the toxin profile produced, as long as the strain contained the genomic domains sxtA1 and sxtA4.
Elucidation of the cell cycle of dinoflagellates is essential to understand the processes leading... more Elucidation of the cell cycle of dinoflagellates is essential to understand the processes leading to their massive proliferations, known as harmful algal blooms. In this study, we used imaging flow cytometry (IFC) to monitor the changes in DNA content and nuclear and cell morphology that occur during clonal growth of the toxic species Alexandrium minutum Halim. Our results indicate that the population was in S phase (C→2C DNA content) during the light period, whereas haploid cells with a C DNA content peaked only during a short interval of the dark period. The timing of the phases, identified based on the nuclear morphology and cytoplasmic-to-nuclear (CNR) ratio of the cells, suggests that the length of the G2/M phase is regulated by nutrient levels whereas the beginning of S phase is clock controlled. In addition we found that up to 7% of individual cells achieved a DNA content higher than 2C, indicative of either zygote formation and replication (homothallism), or of double-haploid cells able to divide (polyploid forms). Cells belonging to different cell cycle phases (G1-S-G2/M) could be readily discriminated based on nuclear size. Our study provides evidence of cell-cycle plasticity during clonal growth and unambiguously characterizes the cell-cycle phases of this dinoflagellate species.
Cell abundances and distributions of Alexandrium catenella resting cysts in recent sediments were... more Cell abundances and distributions of Alexandrium catenella resting cysts in recent sediments were studied along time at two locations in the Chilean Inland Sea exposed to different oceanographic conditions: Low Bay, which is much more open to the ocean than the more interior and protected Ovalada Island. The bloom began in interior areas but maximum cyst concentrations were recorded in locations more open to the ocean, at the end of the Moraleda channel. Our results showed a time lapse of around 3 months from the bloom peak (planktonic population) until the number of resting cysts in the sediments reached a maximum. Three months later, less than 10% of the A. catenella cysts remained in the sediments. Maximum cyst numbers in the water column occurred one month after the planktonic peak, when no cells were present. The dinoflagellate assemblage at both study sites was dominated by heterotrophic cysts, except during the A. catenella bloom. CCA analyses of species composition and environmental factors indicated that the frequency of A. catenella blooms was associated with low temperatures, but not with salinity, chlorophyll a concentration, and predator presence (measured as clam biomass). However, resting cyst distribution was only related to cell abundance and location. The occurrence of A. catenella cysts was also associated with that of cysts from the toxic species Protoceratium reticulatum. By shedding light on the ecological requirements of A. catenella blooms, our observations support the relevance of encystment as a mechanism of bloom termination and show a very fast depletion of cysts from the sediments (<3 months), which suggest a small role for resting cyst deposits in the recurrence of A. catenella blooms in this area.
The dynamics and consequences of host–parasite coevolution depend on the nature of host genotype-... more The dynamics and consequences of host–parasite coevolution depend on the nature of host genotype-by-parasite genotype interactions (G*G) for host and parasite fitness. G*G with crossing reaction norms can yield cyclic dynamics of allele frequencies (“Red Queen” dynamics) while G*G where the
variance among host genotypes differs between parasite genotypes results in selective sweeps (“arms race” dynamics). Here, we investigate the relative potential for arms race and Red Queen coevolution in a protist host–parasite system, the dinoflagellate Alexandrium minutum and its parasite
Parvilucifera sinerae. We challenged nine different clones of
A. minutum with 10 clones of P. sinerae in a fully factorial design and measured infection success and host and parasite fitness. Each host genotype was successfully infected by four to ten of the parasite genotypes. There were strong G*Gs for infection success, as well as both host and parasite fitness. About three quarters of the G*G variance components for host and parasite fitness were due to crossing reaction norms. There were no general costs of resistance or infectivity. We conclude that there is high potential for Red Queen dynamics in this host–parasite system.
Dinoflagellates belonging to the genus Gambierdiscus are the causative agent of ciguatera fish po... more Dinoflagellates belonging to the genus Gambierdiscus are the causative agent of ciguatera fish poisoning (CFP). This syndrome, which is widespread in tropical and subtropical regions, has recently been reported also in temperate latitudes. Taxonomic studies of Gambierdiscus have yet to completely couple the morphological features of member species with their genetics. In this study, the cellular and nuclear morphology of a single strain of one species of Gambierdiscus was determined in cells grown under different culture conditions. The results showed a wide-ranging variability of cell sizes, together with a clear relationship between cell size and nuclear morphology. Thus, small cells were associated with round to oval or slightly U-shaped nuclei and large cells with obviously U-shaped nuclei. Most cells exhibited the typical anterio-posteriorly compressed lenticular, shape of Gambierdiscus, with the exception of a few small globular-shaped specimens. In all cells, regardless of their size, the arrangement of the thecal plates was typical of lenticular Gambierdiscus. Dividing cells were consistently the largest. In these cells, nuclear morphology, karyokinesis, and cytokinesis were characterized. Cells underwent division only during the dark period, thus demonstrating their spontaneous synchronized division. Cellular forms related to the sexual cycle were also present in the cultures and included gamete pairs and putative meiotic planozygotes. The effect of the culture medium was studied by means of principal component analyses, which showed a positive correlation between the medium used and nuclear size and shape but not cell size.
Alexandrium ostenfeldii is present in a wide variety of environments in coastal areas worldwide a... more Alexandrium ostenfeldii is present in a wide variety of environments in coastal areas worldwide and is the only dinoflagellate known species that produces paralytic shellfish poisoning (PSP) toxins and two types of cyclic imines, spirolides (SPXs) and gymnodimines (GYMs). The increasing frequency of A. ostenfeldii blooms in the Baltic Sea has been attributed to the warming water in this region. To learn more about the optimal environmental conditions favoring the proliferation of A. ostenfeldii and its complex toxicity, the effects of temperature and salinity on the kinetics of both the growth and the net toxin production of this species were examined using a factorial design and a response-surface analysis (RSA). The results showed that the growth of Baltic A. ostenfeldii occurs over a wide range of temperatures and salinities (12.5–25.5°C and 5–21, respectively), with optimal growth conditions achieved at a temperature of 25.5°C and a salinity of 11.2. Together with the finding that a salinity > 21 was the only growth-limiting factor detected for this strain, this study provides important insights into the autecology and population distribution of this species in the Baltic Sea. The presence of PSP toxins, including gonyautoxin (GTX)-3, GTX-2, and saxitoxin (STX), and GYMs (GYM-A and GYM-B/-C analogues) was detected under all temperature and salinity conditions tested and in the majority of the cases was concomitant with both the exponential growth and stationary phases of the dinoflagellate’s growth cycle. Toxin concentrations were maximal at temperatures and salinities of 20.9°C and 17 for the GYM-A analogue and > 19°C and 15 for PSP toxins, respectively. The ecological implications of the optimal conditions for growth and toxin production of A. ostenfeldii in the Baltic Sea are discussed.
Dinoflagellates are haploid eukaryotic microalgae in which rapid proliferation causes dense bloom... more Dinoflagellates are haploid eukaryotic microalgae in which rapid proliferation causes dense blooms, with harmful health and economic effects to humans. The proliferation mode is mainly asexual, as the sexual cycle is believed to be rare and restricted to stressful environmental conditions. However, sexuality is key to explaining the recurrence of many dinoflagellate blooms because in many species the fate of the planktonic zygotes (planozygotes) is the formation of resistant cysts in the seabed (encystment). Nevertheless, recent research has shown that individually isolated planozygotes in the lab can enter other routes besides encystment, a behavior of which the relevance has not been explored at the population level. In this study, using imaging flow cytometry, cell sorting, and Fluorescence In Situ Hybridization (FISH), we followed DNA content and nuclear changes in a population of the toxic dinoflagellate Alexandrium minutum that was induced to encystment. Our results first show that planozygotes behave like a population with an “encystment-independent” division cycle, which is light-controlled and follows the same Light:Dark (L:D) pattern as the cycle governing the haploid mitosis. Resting cyst formation was the fate of just a small fraction of the planozygotes formed and was restricted to a period of strongly limited nutrient conditions. The diploid-haploid turnover between L:D cycles was consistent with two-step meiosis. However, the diel and morphological division pattern of the planozygote division also suggests mitosis, which would imply that this species is not haplontic, as previously considered, but biphasic, because individuals could undergo mitotic divisions in both the sexual (diploid) and the asexual (haploid) phases. We also report incomplete genome duplication processes. Our work calls for a reconsideration of the dogma of rare sex in dinoflagellates.
Parasites exert important top-down control of their host populations. The host-parasite system fo... more Parasites exert important top-down control of their host populations. The host-parasite system formed by Alexandrium minutum (Dinophyceae) and Parvilucifera sinerae (Perkinsozoa) offers an opportunity to advance our knowledge of parasitism in planktonic communities. In this study, DNA extracted from 73 clonal strains of P. sinerae, from 10 different locations along the Atlantic and Mediterranean coasts, was used to genetically characterize this parasitoid at the species level. All strains showed identical sequences of the small and large subunits and internal transcribed spacer of the ribosomal RNA, as well as of the β-tubulin genes. However, the phenotypical characterization showed variability in terms of host invasion, zoospore success, maturation time, half-maximal infection, and infection rate. This characterization grouped the strains within 3 phenotypic types distinguished by virulence traits. A particular virulence pattern could not be ascribed to host-cell bloom appearance or to the location or year of parasite-strain isolation; rather, some parasitoid strains from the same bloom significantly differed in their virulence traits. Identical markers such as ITS and β-tubulin genes of P. sinerae strains from different geographic areas and from different years precludes their use in assessing intra-specific diversity and could indicate a recent dispersion of this species.
Abstract
Background: Marine microbial protists, in particular, dinoflagellates, produce polyketi... more Abstract
Background: Marine microbial protists, in particular, dinoflagellates, produce polyketide toxins with ecosystem-wide and human health impacts. Species of Gambierdiscus produce the polyether ladder compounds ciguatoxins and maitotoxins,
which can lead to ciguatera fish poisoning, a serious human illness associated with reef fish consumption. Genes
associated with the biosynthesis of polyether ladder compounds are yet to be elucidated, however, stable isotope feeding studies of such compounds consistently support their polyketide origin indicating that polyketide synthases are
involved in their biosynthesis.
Results: Here, we report the toxicity, genome size, gene content and transcriptome of Gambierdiscus australes and G.
belizeanus. G. australes produced maitotoxin-1 and maitotoxin-3, while G. belizeanus produced maitotoxin-3, for which cell extracts were toxic to mice by IP injection (LD50 = 3.8 mg kg-1). The gene catalogues comprised 83,353 and 84,870
unique contigs, with genome sizes of 32.5 ± 3.7 Gbp and 35 ± 0.88 Gbp, respectively, and are amongst the most
comprehensive yet reported from a dinoflagellate. We found three hundred and six genes involved in polyketide
biosynthesis, including one hundred and ninty-two ketoacyl synthase transcripts, which formed five unique
phylogenetic clusters.
Conclusions: Two clusters were unique to these maitotoxin-producing dinoflagellate species, suggesting that they may
be associated with maitotoxin biosynthesis. This work represents a significant step forward in our understanding of the genetic basis of polyketide production in dinoflagellates, in particular, species responsible for ciguatera fish poisoning.
Dinoflagellates are microscopic aquatic eukaryotes with huge genomes and an unusual cell regulati... more Dinoflagellates are microscopic aquatic eukaryotes with huge genomes and an unusual cell regulation. For example, most genes are present in numerous copies and all copies seem to be obligatorily transcribed. The consequence of the gene copy number for final protein synthesis is however not clear. One such gene is sxtA, the starting gene of paralytic shellfish toxin (PST) synthesis. PSTs are small neurotoxic compounds that can accumulate in the food chain and cause serious poisoning incidences when ingested. They are produced by dinoflagellates of the genera Alexandrium, Gymnodium and Pyrodinium.
Here we investigated if the genomic copy number of sxtA4 is related to PST content in Alexandrium minutum cells. SxtA4 is the 4th domain of the sxtA gene and its presence is essential for PST synthesis in dinoflagellates. We used PST and genome size measurements as well as quantitative PCR to analyze sxtA4 copy number and toxin content in 15 A. minutum strains. Our results show a strong positive correlation between the sxtA4 copy number and the total amount of PST produced in actively growing A. minutum cells. This correlation was independent of the toxin profile produced, as long as the strain contained the genomic domains sxtA1 and sxtA4.
Elucidation of the cell cycle of dinoflagellates is essential to understand the processes leading... more Elucidation of the cell cycle of dinoflagellates is essential to understand the processes leading to their massive proliferations, known as harmful algal blooms. In this study, we used imaging flow cytometry (IFC) to monitor the changes in DNA content and nuclear and cell morphology that occur during clonal growth of the toxic species Alexandrium minutum Halim. Our results indicate that the population was in S phase (C→2C DNA content) during the light period, whereas haploid cells with a C DNA content peaked only during a short interval of the dark period. The timing of the phases, identified based on the nuclear morphology and cytoplasmic-to-nuclear (CNR) ratio of the cells, suggests that the length of the G2/M phase is regulated by nutrient levels whereas the beginning of S phase is clock controlled. In addition we found that up to 7% of individual cells achieved a DNA content higher than 2C, indicative of either zygote formation and replication (homothallism), or of double-haploid cells able to divide (polyploid forms). Cells belonging to different cell cycle phases (G1-S-G2/M) could be readily discriminated based on nuclear size. Our study provides evidence of cell-cycle plasticity during clonal growth and unambiguously characterizes the cell-cycle phases of this dinoflagellate species.
Cell abundances and distributions of Alexandrium catenella resting cysts in recent sediments were... more Cell abundances and distributions of Alexandrium catenella resting cysts in recent sediments were studied along time at two locations in the Chilean Inland Sea exposed to different oceanographic conditions: Low Bay, which is much more open to the ocean than the more interior and protected Ovalada Island. The bloom began in interior areas but maximum cyst concentrations were recorded in locations more open to the ocean, at the end of the Moraleda channel. Our results showed a time lapse of around 3 months from the bloom peak (planktonic population) until the number of resting cysts in the sediments reached a maximum. Three months later, less than 10% of the A. catenella cysts remained in the sediments. Maximum cyst numbers in the water column occurred one month after the planktonic peak, when no cells were present. The dinoflagellate assemblage at both study sites was dominated by heterotrophic cysts, except during the A. catenella bloom. CCA analyses of species composition and environmental factors indicated that the frequency of A. catenella blooms was associated with low temperatures, but not with salinity, chlorophyll a concentration, and predator presence (measured as clam biomass). However, resting cyst distribution was only related to cell abundance and location. The occurrence of A. catenella cysts was also associated with that of cysts from the toxic species Protoceratium reticulatum. By shedding light on the ecological requirements of A. catenella blooms, our observations support the relevance of encystment as a mechanism of bloom termination and show a very fast depletion of cysts from the sediments (<3 months), which suggest a small role for resting cyst deposits in the recurrence of A. catenella blooms in this area.
The dynamics and consequences of host–parasite coevolution depend on the nature of host genotype-... more The dynamics and consequences of host–parasite coevolution depend on the nature of host genotype-by-parasite genotype interactions (G*G) for host and parasite fitness. G*G with crossing reaction norms can yield cyclic dynamics of allele frequencies (“Red Queen” dynamics) while G*G where the
variance among host genotypes differs between parasite genotypes results in selective sweeps (“arms race” dynamics). Here, we investigate the relative potential for arms race and Red Queen coevolution in a protist host–parasite system, the dinoflagellate Alexandrium minutum and its parasite
Parvilucifera sinerae. We challenged nine different clones of
A. minutum with 10 clones of P. sinerae in a fully factorial design and measured infection success and host and parasite fitness. Each host genotype was successfully infected by four to ten of the parasite genotypes. There were strong G*Gs for infection success, as well as both host and parasite fitness. About three quarters of the G*G variance components for host and parasite fitness were due to crossing reaction norms. There were no general costs of resistance or infectivity. We conclude that there is high potential for Red Queen dynamics in this host–parasite system.
Dinoflagellates belonging to the genus Gambierdiscus are the causative agent of ciguatera fish po... more Dinoflagellates belonging to the genus Gambierdiscus are the causative agent of ciguatera fish poisoning (CFP). This syndrome, which is widespread in tropical and subtropical regions, has recently been reported also in temperate latitudes. Taxonomic studies of Gambierdiscus have yet to completely couple the morphological features of member species with their genetics. In this study, the cellular and nuclear morphology of a single strain of one species of Gambierdiscus was determined in cells grown under different culture conditions. The results showed a wide-ranging variability of cell sizes, together with a clear relationship between cell size and nuclear morphology. Thus, small cells were associated with round to oval or slightly U-shaped nuclei and large cells with obviously U-shaped nuclei. Most cells exhibited the typical anterio-posteriorly compressed lenticular, shape of Gambierdiscus, with the exception of a few small globular-shaped specimens. In all cells, regardless of their size, the arrangement of the thecal plates was typical of lenticular Gambierdiscus. Dividing cells were consistently the largest. In these cells, nuclear morphology, karyokinesis, and cytokinesis were characterized. Cells underwent division only during the dark period, thus demonstrating their spontaneous synchronized division. Cellular forms related to the sexual cycle were also present in the cultures and included gamete pairs and putative meiotic planozygotes. The effect of the culture medium was studied by means of principal component analyses, which showed a positive correlation between the medium used and nuclear size and shape but not cell size.
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Papers by Rosa Isabel Figueroa
mainly asexual, as the sexual cycle is believed to be rare and restricted to stressful environmental conditions. However, sexuality is key to explaining the recurrence of many dinoflagellate blooms because in many species the fate of the planktonic zygotes (planozygotes)
is the formation of resistant cysts in the seabed (encystment). Nevertheless, recent research has shown that individually isolated planozygotes in the lab can enter other routes
besides encystment, a behavior of which the relevance has not been explored at the population level. In this study, using imaging flow cytometry, cell sorting, and Fluorescence In
Situ Hybridization (FISH), we followed DNA content and nuclear changes in a population of the toxic dinoflagellate Alexandrium minutum that was induced to encystment. Our results
first show that planozygotes behave like a population with an “encystment-independent” division cycle, which is light-controlled and follows the same Light:Dark (L:D) pattern as the
cycle governing the haploid mitosis. Resting cyst formation was the fate of just a small fraction
of the planozygotes formed and was restricted to a period of strongly limited nutrient conditions. The diploid-haploid turnover between L:D cycles was consistent with two-step
meiosis. However, the diel and morphological division pattern of the planozygote division also suggests mitosis, which would imply that this species is not haplontic, as previously considered, but biphasic, because individuals could undergo mitotic divisions in both the sexual (diploid) and the asexual (haploid) phases. We also report incomplete genome duplication processes. Our work calls for a reconsideration of the dogma of rare sex in dinoflagellates.
Background: Marine microbial protists, in particular, dinoflagellates, produce polyketide toxins with ecosystem-wide and human health impacts. Species of Gambierdiscus produce the polyether ladder compounds ciguatoxins and maitotoxins,
which can lead to ciguatera fish poisoning, a serious human illness associated with reef fish consumption. Genes
associated with the biosynthesis of polyether ladder compounds are yet to be elucidated, however, stable isotope feeding studies of such compounds consistently support their polyketide origin indicating that polyketide synthases are
involved in their biosynthesis.
Results: Here, we report the toxicity, genome size, gene content and transcriptome of Gambierdiscus australes and G.
belizeanus. G. australes produced maitotoxin-1 and maitotoxin-3, while G. belizeanus produced maitotoxin-3, for which cell extracts were toxic to mice by IP injection (LD50 = 3.8 mg kg-1). The gene catalogues comprised 83,353 and 84,870
unique contigs, with genome sizes of 32.5 ± 3.7 Gbp and 35 ± 0.88 Gbp, respectively, and are amongst the most
comprehensive yet reported from a dinoflagellate. We found three hundred and six genes involved in polyketide
biosynthesis, including one hundred and ninty-two ketoacyl synthase transcripts, which formed five unique
phylogenetic clusters.
Conclusions: Two clusters were unique to these maitotoxin-producing dinoflagellate species, suggesting that they may
be associated with maitotoxin biosynthesis. This work represents a significant step forward in our understanding of the genetic basis of polyketide production in dinoflagellates, in particular, species responsible for ciguatera fish poisoning.
Here we investigated if the genomic copy number of sxtA4 is related to PST content in Alexandrium minutum cells. SxtA4 is the 4th domain of the sxtA gene and its presence is essential for PST synthesis in dinoflagellates. We used PST and genome size measurements as well as quantitative PCR to analyze sxtA4 copy number and toxin content in 15 A. minutum strains. Our results show a strong positive correlation between the sxtA4 copy number and the total amount of PST produced in actively growing A. minutum cells. This correlation was independent of the toxin profile produced, as long as the strain contained the genomic domains sxtA1 and sxtA4.
variance among host genotypes differs between parasite genotypes results in selective sweeps (“arms race” dynamics). Here, we investigate the relative potential for arms race and Red Queen coevolution in a protist host–parasite system, the dinoflagellate Alexandrium minutum and its parasite
Parvilucifera sinerae. We challenged nine different clones of
A. minutum with 10 clones of P. sinerae in a fully factorial design and measured infection success and host and parasite fitness. Each host genotype was successfully infected by four to ten of the parasite genotypes. There were strong G*Gs for infection success, as well as both host and parasite fitness. About three quarters of the G*G variance components for host and parasite fitness were due to crossing reaction norms. There were no general costs of resistance or infectivity. We conclude that there is high potential for Red Queen dynamics in this host–parasite system.
mainly asexual, as the sexual cycle is believed to be rare and restricted to stressful environmental conditions. However, sexuality is key to explaining the recurrence of many dinoflagellate blooms because in many species the fate of the planktonic zygotes (planozygotes)
is the formation of resistant cysts in the seabed (encystment). Nevertheless, recent research has shown that individually isolated planozygotes in the lab can enter other routes
besides encystment, a behavior of which the relevance has not been explored at the population level. In this study, using imaging flow cytometry, cell sorting, and Fluorescence In
Situ Hybridization (FISH), we followed DNA content and nuclear changes in a population of the toxic dinoflagellate Alexandrium minutum that was induced to encystment. Our results
first show that planozygotes behave like a population with an “encystment-independent” division cycle, which is light-controlled and follows the same Light:Dark (L:D) pattern as the
cycle governing the haploid mitosis. Resting cyst formation was the fate of just a small fraction
of the planozygotes formed and was restricted to a period of strongly limited nutrient conditions. The diploid-haploid turnover between L:D cycles was consistent with two-step
meiosis. However, the diel and morphological division pattern of the planozygote division also suggests mitosis, which would imply that this species is not haplontic, as previously considered, but biphasic, because individuals could undergo mitotic divisions in both the sexual (diploid) and the asexual (haploid) phases. We also report incomplete genome duplication processes. Our work calls for a reconsideration of the dogma of rare sex in dinoflagellates.
Background: Marine microbial protists, in particular, dinoflagellates, produce polyketide toxins with ecosystem-wide and human health impacts. Species of Gambierdiscus produce the polyether ladder compounds ciguatoxins and maitotoxins,
which can lead to ciguatera fish poisoning, a serious human illness associated with reef fish consumption. Genes
associated with the biosynthesis of polyether ladder compounds are yet to be elucidated, however, stable isotope feeding studies of such compounds consistently support their polyketide origin indicating that polyketide synthases are
involved in their biosynthesis.
Results: Here, we report the toxicity, genome size, gene content and transcriptome of Gambierdiscus australes and G.
belizeanus. G. australes produced maitotoxin-1 and maitotoxin-3, while G. belizeanus produced maitotoxin-3, for which cell extracts were toxic to mice by IP injection (LD50 = 3.8 mg kg-1). The gene catalogues comprised 83,353 and 84,870
unique contigs, with genome sizes of 32.5 ± 3.7 Gbp and 35 ± 0.88 Gbp, respectively, and are amongst the most
comprehensive yet reported from a dinoflagellate. We found three hundred and six genes involved in polyketide
biosynthesis, including one hundred and ninty-two ketoacyl synthase transcripts, which formed five unique
phylogenetic clusters.
Conclusions: Two clusters were unique to these maitotoxin-producing dinoflagellate species, suggesting that they may
be associated with maitotoxin biosynthesis. This work represents a significant step forward in our understanding of the genetic basis of polyketide production in dinoflagellates, in particular, species responsible for ciguatera fish poisoning.
Here we investigated if the genomic copy number of sxtA4 is related to PST content in Alexandrium minutum cells. SxtA4 is the 4th domain of the sxtA gene and its presence is essential for PST synthesis in dinoflagellates. We used PST and genome size measurements as well as quantitative PCR to analyze sxtA4 copy number and toxin content in 15 A. minutum strains. Our results show a strong positive correlation between the sxtA4 copy number and the total amount of PST produced in actively growing A. minutum cells. This correlation was independent of the toxin profile produced, as long as the strain contained the genomic domains sxtA1 and sxtA4.
variance among host genotypes differs between parasite genotypes results in selective sweeps (“arms race” dynamics). Here, we investigate the relative potential for arms race and Red Queen coevolution in a protist host–parasite system, the dinoflagellate Alexandrium minutum and its parasite
Parvilucifera sinerae. We challenged nine different clones of
A. minutum with 10 clones of P. sinerae in a fully factorial design and measured infection success and host and parasite fitness. Each host genotype was successfully infected by four to ten of the parasite genotypes. There were strong G*Gs for infection success, as well as both host and parasite fitness. About three quarters of the G*G variance components for host and parasite fitness were due to crossing reaction norms. There were no general costs of resistance or infectivity. We conclude that there is high potential for Red Queen dynamics in this host–parasite system.