Karin Rengefors

Allelopathy in phytoplankton - biochemical, ecological and evolutionary aspects

Abstract It is considered self-evident that chemical interactions are a component of competition in terrestrial systems, but they are largely unknown in aquatic systems. In this review, we propose that chemical interactions, specifically allelopathy, are an important part of phytoplankton competition. Allelopathy, as defined here, applies only to the inhibitory effects of secondary metabolites produced by one species on the growth or physiological function of another phytoplankton species. A number of approaches are used to study allelopathy, but there is no standard methodology available. One of the methods used is cross-culturing, in which the cell-free filtrate of a donor alga is added to the medium of the target species. Another is to study the effect of cell extracts of unknown constituents, isolated exudates or purified allelochemicals on the growth of other algal species. There is a clear lack of controlled field experiments because few allelochemicals have been identified. Molecular methods will be important in future to study the expression and regulation of allelochemicals. Most of the identified allelochemicals have been described for cyanobacteria but some known toxins marine dinoflagellates and freshwater cyanobacteria also have an allelochemical effect. The mode of action of allelochemicals spans a wide range. The most common effect is to cause cell lysis, blistering, or growth inhibition. The factors that affect allelochemical production have not been studied much, although nutrient limitation, pH, and temperature appear to have an effect. The evolutionary aspects of allelopathy remain largely unknown, but we hypothesize that the producers of allelochemicals should gain a competitive advantage over other phytoplankton. Finally, we discuss the possibility of using allelochemicals to combat harmful algal blooms (HABs). Allelopathic agents are used for biological control in agriculture, e.g. green manures to control soil diseases in Australia, but they have not yet been applied in the context of HABs. We suggest that phytoplankton allelochemicals have the potential for management of HABs in localized areas.

The Marine Microbial Eukaryote Transcriptome Sequencing Project (MMETSP): Illuminating the Functional Diversity of Eukaryotic Life in the Oceans through Transcriptome Sequencing.

Current sampling of genomic sequence data from eukaryotes is relatively poor, biased, and inadequate to address important questions about their biology, evolution, and ecology; this Community Page describes a resource of 700 transcriptomes from marine microbial eukaryotes to help understand their role in the worlds oceans.

Infrequent marine-freshwater transitions in the microbial world

Until recently, the evolutionary relationships between marine and freshwater microbes were unclear, but the use of molecular phylogenies is beginning to shed light on this subject. An increasing amount of studies are showing that marine and freshwater microbes (including viruses) are usually not closely related, often grouping into distinct marine and freshwater phylogenetic clusters, similar to what has been reported before for macroorganisms. These studies indicate that marine-freshwater transitions have been infrequent events during the diversification of microbes and that most of these transitions occurred a long time ago in evolutionary terms. Here we discuss the significance of recent studies addressing this question and consider possible avenues for future research.

Biogeography of bacterial communities exposed to progressive long-term environmental change

The response of microbial communities to long-term environmental change is poorly understood. Here, we study bacterioplankton communities in a unique system of coastal Antarctic lakes that were exposed to progressive long-term environmental change, using 454 pyrosequencing of the 16S rDNA gene (V3–V4 regions). At the time of formation, most of the studied lakes harbored marine-coastal microbial communities, as they were connected to the sea. During the past 20 000 years, most lakes isolated from the sea, and subsequently they experienced a gradual, but strong, salinity change that eventually developed into a gradient ranging from freshwater (salinity 0) to hypersaline (salinity 100). Our results indicated that present bacterioplankton community composition was strongly correlated with salinity and weakly correlated with geographical distance between lakes. A few abundant taxa were shared between some lakes and coastal marine communities. Nevertheless, lakes contained a large number of taxa that were not detected in the adjacent sea. Abundant and rare taxa within saline communities presented similar biogeography, suggesting that these groups have comparable environmental sensitivity. Habitat specialists and generalists were detected among abundant and rare taxa, with specialists being relatively more abundant at the extremes of the salinity gradient. Altogether, progressive long-term salinity change appears to have promoted the diversification of bacterioplankton communities by modifying the composition of ancestral communities and by allowing the establishment of new taxa.

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

Adaptations and counteradaptations are common in coevolving predator- prey 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 Daphnias 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 magnaa 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 individuals 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.

Algal cyst dormancy: a temporal escape from herbivory

Many phytoplankton species form resting cysts and remain dormant for part of the year. The subsequent excystment is regulated by the external environment and internal maturation processes. Here we assessed the excystment of the dinoflagellates Ceratium hirundinella and Peridinium aciculiferum in relation to herbivores and temperature in laboratory and field studies. C. hirundinella, which has a grazer–resistant morphology, forms summer blooms, whereas P. aciculiferum, which is vulnerable to grazers, grows underneath the ice during winter. In our study, herbivore abundance, and thereby grazing pressure, was low during periods when water temperatures were low, and the abundance of P. aciculiferum was high. In the laboratory experiment, excystment of C. hirundinella occurred at high temperatures irrespective of whether zooplankton exudate was added or not, whereas at intermediate temperatures, excystment was lower if zooplankton exudate was added. Germination of P. aciculiferum cysts was lower in the presence of exudate from a zooplankton culture than in controls at all temperatures. Our studies suggest that dinoflagellates use the presence of zooplankton in addition to temperature as a cue to determine when to excyst. Consequently, not only abiotic factors, but also the composition of the food web, may determine succession and composition of phytoplankton communities.

Phenotypically different microalgal morphospecies with identical ribosomal DNA: a case of rapid adaptive evolution?

The agents driving the divergence and speciation of free-living microbial populations are still largely unknown. We investigated the dinoflagellate morphospecies Scrippsiella hangoei and Peridinium aciculiferum, which abound in the Baltic Sea and in northern temperate lakes, respectively. Electron microscopy analyses showed significant interspecific differences in the external cellular morphology, but a similar plate pattern in the characteristic dinoflagellate armor. Experimentally, S. hangoei grew in a wide range of salinities (0–30), whereas P. aciculiferum only grew in low salinities (0–3). Despite these phenotypic differences and the habitat segregation, molecular analyses showed identical ribosomal DNA sequences (ITS1, ITS2, 5.8S, SSU, and partial LSU) for both morphospecies. Yet, a strong interspecific genetic isolation was indicated by amplified fragment length polymorphism (FST = 0.76) and cytochrome b (cob) sequence divergence (∼1.90%). Phylogenetic reconstructions based on ribosomal (SSU, LSU) and mitochondrial (cob) DNA indicated a recent marine ancestor for P. aciculiferum. In conclusion, we suggest that the lacustrine P. aciculiferum and the marine-brackish S. hangoei diverged very recently, after a marine–freshwater transition that exposed the ancestral populations to different selective pressures. This hypothetical scenario agrees with mounting data indicating a significant role of natural selection in the divergence of free-living microbes, despite their virtually unrestricted dispersal capabilities. Finally, our results indicate that identical ITS rDNA sequences do not necessarily imply the same microbial species, as commonly assumed.

ENVIRONMENTAL AND ENDOGENOUS REGULATION OF CYST GERMINATION IN TWO FRESHWATER DINOFLAGELLATES

The role of excystment in relation to seasonal succession was investigated in two freshwater dinoflagellates, Ceratium hirundinella (O.F. Müller) Dujardin and Peridinium aciculiferum (Lemmermann). Field studies and laboratory experiments were performed to determine which factors regulate the timing of cyst germination. Environmental factors (temperature, light, nutrients, and anoxia) and endogenous factors (maturation period and biological clock) were investigated. Our main results indicate that temperature and internal maturation period determine when germination can occur. C. hirundinella had a maturation period of 4.5 months and germinated in the laboratory and in the field at temperatures above 6° C. P. aciculiferum had a maturation period of 2.5 months and germinated in the laboratory and in the field at temperatures below 7° C. In addition, our results indicated that both species were regulated by a biological clock. Furthermore, anoxia prevented the germination of C. hirundinella, contrary to results in earlier studies. To conclude, we could explain the appearance in plankton of the two dinoflagellate species through two main factors regulating excystment, that is, temperature and maturation period.

Life cyle and sexuality of the freshwater raphidophyte Gonyostomum semen (Raphidophyceae)

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.

Genetic Diversity Patterns in Five Protist Species Occurring in Lakes

Little is known about the extent of the genetic diversity and its structuring patterns in protist species living in lakes. Here, we have investigated the genetic diversity patterns within five dinoflagellate species (Peridinium aciculiferum, Peridinium cinctum, Peridiniopsis borgei, Polarella glacialis, Scrippsiella aff. hangoei) that are present in lakes and sometimes, in marine habitats located in polar and temperate regions. A total of 68 clonal strains were investigated using Amplified Fragment Length Polymorphism (AFLP), a sensitive genetic fingerprinting technique. All used strains within each species had identical ITS nuclear ribosomal DNA sequences, a characteristic that indicates that they likely belong to the same species. We found a wide variability in the genetic diversity among species (between 20% and 90% of polymorphic loci; Neis gene diversity between 0.08 and 0.37). In some cases, our analyses suggested the presence of different genetically homogeneous subgroups (genetic populations) within the same water body. Thus, it appears that different genetic populations can coexist within the same lake despite the likely occurrence of recombination that tends to homogenize the gene pool. Overall, our results indicated that a large number of dinoflagellate genotypes are present in lake populations, instead of a few dominating ones. In addition, our study shows that protists with identical ITS sequences can harbor considerable amounts of genetic diversity.