Single-cell genomics of a bloom-forming phytoplankton species reveals population genetic structure across continents

The study of microbial diversity over time and space is fundamental to the understanding of their ecology and evolution. The underlying processes driving these patterns are not fully resolved but can be studied using population genetic approaches. Here we investigated the population genetic structure of Gonyostomum semen, a bloom-forming phytoplankton species, across two continents. The species appears to be expanding in Europe, whereas similar trends are not observed in the United States. Our aim was to investigate if populations of G. semen in Europe and in the United States are genetically differentiated, if there is population genetic structure within the continents, and what the potential drivers of differentiation are. To this end, we used a novel method based on Single Amplified Genomes (SAGs) combined with RADseq (SAG-RAD), that allows de novo genotyping of natural single cell isolates without the need for culturing. We amplified over 900 single-cell genomes from 25 lake populations across Europe and the United States and identified two distinct population clusters, one in Europe and another in the United States. Low genetic diversity in European populations supports the hypothesized recent expansion of G. semen on this continent. Geographic population structure within each continent was associated with differences in environmental variables which may have led to ecological divergence of population clusters. Overall, our results show that SAG-RAD can be used to analyze microalgal population structure and differentiation based on single-cell isolates from natural, uncultured samples

Increased consumer fitness following transfer of toxin tolerance to offspring via maternal effects

Adaptations and, counteradaptations are common in coevolving predatorprey systems, but little is known of the role of maternal transfer of adaptive traits in mediating species interactions. Here, we focused on tolerance against cyanobacterial toxins and asked whether this tolerance was an induced defense developed during Daphnia's lifetime, whether it was a trait that is constantly expressed, and whether such tolerance to the toxin can be transferred to the next generation through maternal effects. These questions were addressed by feeding a single clone of Daphnia magna a diet with and without algal toxin and recording changes in fitness (as intrinsic rate of population increase). Analysis of F1, F2, and F3 generations revealed that the increased tolerance to toxic Microcystis was an inducible defense developed during an individual's lifetime, and that this trait could be transferred from mother to offspring. This maternal effect was expressed in several fitness parameters, including shorter time to maturity and first reproduction, and higher numbers of offspring compared to inexperienced individuals. In some circumstances, such maternal effects may increase population production by up to 40% and may help to stabilize material and energy transfer to higher trophic levels

Priority effects in a planktonic bloom-forming marine diatom

Priority effects occur when a species or genotype with earlier arrival has an advantage such that its relative abundance in the community or population is increased compared with later-arriving species. Few studies have dealt with this concept in the context of within-species competition. Skeletonema marinoi is a marine diatom that shows a high degree of genetic differentiation between populations over small geographical distances. To test whether historical events such as priority effects may have been important in inducing these patterns of population differentiation, we performed microcosm experiments with successive inoculation of different S. marinoi strains. Our results show that even in the absence of a numerical advantage, significant priority effects were evident. We propose that priority effects may be an important mechanism in initiating population genetic differentiation

On the control of HAB species using low biosurfactant concentrations

Biosurfactants have been suggested as a method to control harmful algal blooms (HABs), but warrant further and more in-depth investigation. Here we have investigated the algicidal effect of a biosurfactant produced by the bacterium Pseudomonas aeruginosa on five diverse marine and freshwater HAB species that have not been tested previously. These include Alexandrium minutum (Dinophycaee), Karenia brevis (Dinophyceae), Pseudonitzschia sp. (Bacillariophyceae), in marine ecosystems, and Gonyostomum semen (Raphidophyceae) and Microcystis aeruginosa (Cyanophyecae) in freshwater. We examined not only lethal but also sub-lethal effects of the biosurfactant. In addition, the effect of the biosurfactant on Daphnia was tested. Our conclusions were that very low biosurfactant concentrations (5 μg mL−1) decreased both the photosynthesis efficiency and the cell viability and that higher concentrations (50 μg mL−1) had lethal effects in four of the five HAB species tested. The low concentrations employed in this study and the diversity of HAB genera tested suggest that biosurfactants may be used to either control initial algal blooms without causing negative side effect to the ecosystem, or to provoke lethal effects when necessary.Postprin

Genetic diversity within and genetic differentiation between blooms of a microalgal species

The field of genetic diversity in protists, particularly phytoplankton, is under expansion. However, little is known regarding variation in genetic diversity within populations over time. The aim of our study was to investigate intrapopulation genetic diversity and genetic differentiation in the freshwater bloom-forming microalga Gonyostomum semen (Raphidophyceae). The study covered a 2-year period including all phases of the bloom. Amplified fragment length polymorphism (AFLP) was used to determine the genetic structure and diversity of the population. Our results showed a significant differentiation between samples collected during the two blooms from consecutive years. Also, an increase of gene diversity and a loss of differentiation among sampling dates were observed over time within a single bloom. The latter observations may reflect the continuous germination of cysts from the sediment. The life cycle characteristics of G. semen, particularly reproduction and recruitment, most likely explain a high proportion of the observed variation. This study highlights the importance of the life cycle for the intraspecific genetic diversity of microbial species, which alternates between sexual and asexual reproduction.Postprin

Grazing resistance allows bloom formation and may explain invasion success of Gonyostomum semen

The nuisance alga Gonyostomum semen (Raphidophyceae) has expanded in the Nordic countries during the last decades and can dominate lake phytoplankton communities almost completely. A possible explanation to its dominance could be limited grazing by zooplankton. We investigated the potential grazing pressure on G. semen using an experimental approach supported by field data. We determined the grazing rate by cladocerans, calanoid copepods, and Chaoborus larvae to determine which were able to feed on G. semen. Only the large cladoceran Daphnia magna was able to feed successfully on G. semen. The large cell size of G. semen was likely a limiting factor for the filtering apparatus of smaller cladocerans. The copepod Eudiaptomus gracilis did not graze on G. semen, although the mechanism behind this selective feeding is still unknown. In addition to the experimental study, we quantified the zooplankton and phytoplankton communities in 40 lakes to determine the composition and abundance of the zooplankton communities co-occurring with G. semen, suggesting that large cladoceran species were not present in lakes where G. semen occurred. Hence, the growth of G. semen is not significantly controlled by grazing in natural systems, which likely facilitates bloom formation and invasion success of G. semen

Seasonal genotype dynamics of a marine dinoflagellate : Pelagic populations are homogeneous and as diverse as benthic seed banks

Genetic diversity is the basis for evolutionary adaptation and selection under changing environmental conditions. Phytoplankton populations are genotypically diverse, can become genetically differentiated within small spatiotemporal scales and many species form resting stages. Resting stage accumulations in sediments (seed banks) are expected to serve as reservoirs for genetic information, but so far their role in maintaining phytoplankton diversity and in evolution has remained unclear. In this study we used the toxic dinoflagellate Alexandrium ostenfeldii (Dinophyceae) as a model organism to investigate if (i) the benthic seed bank is more diverse than the pelagic population and (ii) the pelagic population is seasonally differentiated. Resting stages (benthic) and plankton (pelagic) samples were collected at a coastal bloom site in the Baltic Sea, followed by cell isolation and genotyping using microsatellite markers (MS) and restriction site associated DNA sequencing (RAD). High clonal diversity (98%-100%) combined with intermediate to low gene diversity (0.58-0.03, depending on the marker) was found. Surprisingly, the benthic and pelagic fractions of the population were equally diverse, and the pelagic fraction was temporally homogeneous, despite seasonal fluctuation of environmental selection pressures. The results of this study suggest that continuous benthic-pelagic coupling, combined with frequent sexual reproduction, as indicated by persistent linkage equilibrium, prevent the dominance of single clonal lineages in a dynamic environment. Both processes harmonize the pelagic with the benthic population and thus prevent seasonal population differentiation. At the same time, frequent sexual reproduction and benthic-pelagic coupling maintain high clonal diversity in both habitats.Peer reviewe

Experimental investigation of taxon-specific response of alkaline phosphatase activity in natural freshwater phytoplankton

It is widely accepted that alkaline phosphatase activity (APA) is an efficient indicator of phosphate limitation in freshwater phytoplankton communities. In this study, we investigated whether the response in APA to phosphate limitation differs among the taxa in a mixed phytoplankton assemblage. We used the new enzyme-labeled fluorescence (ELF) technique, which allows microscopic detection of phosphate limitation in individual cells of multiple species. The most prominent findings of this study were that alkaline phosphatase (AP) was induced in many, but not all taxa and that different taxa, as well as different cells within a single taxon, experienced different degrees of phosphate stress under the same environmental conditions. Our approach was to manipulate the limiting nutrient in a natural freshwater phytoplankton community by incubating lake water in the laboratory. We induced nitrogen (N) or phosphate limitation through additions of inorganic nutrients. Both the ELF assay and bulk APA indicated that the lake phytoplankton were not phosphate limited at the start of the experiment. During the experiment, several chlorophyte taxa (e.g., Eudorina and an unidentified solitary spiny coccoid) were driven to phosphate limitation when inorganic N was added, as evidenced by a higher percentage of ELF-labeled cells relative to controls, whereas other chlorophyte taxa such as Actinastrum and Dicryosphaerium were not phosphate stressed under these conditions. In the phosphate-limited treatments, little or no ELF labeling was observed in any cyanobacterial taxa. Furthermore, all taxa observed after the ELF labeling procedure (>10-mum fraction) were labeled with ELF at least on one occasion, demonstrating the wide applicability of the ELF method. By using ELF labeling in tandem with bulk APA, the resolution and analysis of phosphate limitation was increased, allowing the identification of specific phosphate-stressed taxa

Temporal and spatial variability of dissolved organic and inorganic phosphorus, and metrics of phosphorus bioavailability in an upwelling-dominated coastal system

Author Posting. © American Geophysical Union, 2005. This article is posted here by permission of American Geophysical Union for personal use, not for redistribution. The definitive version was published in Journal of Geophysical Research 110 (2005): C10S13, doi:10.1029/2004JC002837.High-frequency temporal and spatial shifts in the various dissolved P pools (total, inorganic, and organic) are linked to upwelling/relaxation events and to phytoplankton bloom dynamics in the upwelling-dominated Oregon coastal system. The presence and regulation of alkaline phosphatase activity (APA) is apparent in the bulk phytoplankton population and in studies of cell-specific APA using Enzyme Labeled Fluorescence (ELF®). Spatial and temporal variability are also evident in phytoplankton community composition and in APA. The spatial pattern of dissolved phosphorus and APA variability can be explained by bottom-controlled patterns of upwelling, and flushing times of different regions within the study area. The presence of APA in eukaryotic taxa indicates that dissolved organic phosphorus (DOP) may contribute to phytoplankton P nutrition in this system, highlighting the need for a more complete understanding of P cycling and bioavailability in the coastal ocean.KCR acknowledges WHOI for rapid-response funding that made possible participation on this first COAST cruise, and NSF-OCE grant 0119134 for support of subsequent work on these and other COAST samples

Towards an ecological understanding of dinoflagellate cyst functions

The life cycle of many dinoflagellates includes at least one nonflagellated benthic stage (cyst). In the literature, the different types of dinoflagellate cysts are mainly defined based on morphological (number and type of layers in the cell wall) and functional (long- or short-term endurance) differences. These characteristics were initially thought to clearly distinguish pellicle (thin-walled) cysts from resting (double-walled) dinoflagellate cysts. The former were considered short-term (temporal) and the latter long-term (resting) cysts. However, during the last two decades further knowledge has highlighted the great intricacy of dinoflagellate life histories, the ecological significance of cyst stages, and the need to clarify the functional and morphological complexities of the different cyst types. Here we review and, when necessary, redefine the concepts of resting and pellicle cysts, examining both their structural and their functional characteristics in the context of the life cycle strategies of several dinoflagellate species.Versión del editor