Genotypic diversity and differentiation among populations of two benthic freshwater diatoms as revealed by microsatellites.

Given their large population sizes and presumed high dispersal capacity, protists are expected to exhibit homogeneous population structure over large spatial scales. On the other hand, the fragmented and short-lived nature of the lentic freshwater habitats that many protists inhabit promotes strong population differentiation. We used microsatellites in two benthic freshwater diatoms, Eunotia bilunaris 'robust' and Sellaphora capitata, sampled from within a pond and connected ponds, through isolated ponds from the same region to western Europe to determine the spatial scale at which differentiation appears. Because periods of low genotypic diversity contribute to population differentiation, we also assessed genotypic diversity. While genotypic diversity was very high to maximal in most samples of both species, some had a markedly lower diversity, with up to half (Eunotia) and over 90% (Sellaphora) of the strains having the same multilocus genotype. Population differentiation showed an isolation-by-distance pattern with very low standardized FST values between samples from the same or connected ponds but high values between isolated ponds, even when situated in the same region. Partial rbcL sequences in Eunotia were consistent with this pattern as isolated ponds in the same region could differ widely in haplotype composition. Populations identified by Structure corresponded to the source ponds, confirming that 'pond' is the main factor structuring these populations. We conclude that freshwater benthic diatom populations are highly fragmented on a regional scale, reflecting either less dispersal than is often assumed or reduced establishment success of immigrants, so that dispersal does not translate into gene flow.

Sulphur levels in saliva as an estimation of sulphur status in cattle: a validation study.

Effective assessment of sulphur (S) status in cattle is important for optimal health, yet remains difficult. Rumen fluid S concentrations are preferred, but difficult to sample under practical conditions. This study aimed to evaluate salivary S concentration as estimator of S status in cattle. Saliva and rumen fluid samples were collected from dairy cows (n = 16) as well as samples of different feedstuffs offered to the animals. The N and S concentrations were determined using the Dumas technique. The average dietary N and S content were calculated as well as N:S ratio of saliva, rumen fluid and diet. Salivary S concentrations could not predict rumen fluid or dietary S concentrations (p > 0.05). The log transformed salivary N:S ratio (x) could predict the rumen fluid N:S ratio (y) with a linear equation of y = 9.83 (±4.59) x + 0.39 (±4.56) (r = 0.497, p = 0.05), but left too much residual variation to serve as reliable predictor. Further research should investigate this relationship in the search for an S status estimator.

Lack of phylogeographic structure in the freshwater cyanobacterium Microcystis aeruginosa suggests global dispersal.

BACKGROUND: Free-living microorganisms have long been assumed to have ubiquitous distributions with little biogeographic signature because they typically exhibit high dispersal potential and large population sizes. However, molecular data provide contrasting results and it is far from clear to what extent dispersal limitation determines geographic structuring of microbial populations. We aimed to determine biogeographical patterns of the bloom-forming freshwater cyanobacterium Microcystis aeruginosa. Being widely distributed on a global scale but patchily on a regional scale, this prokaryote is an ideal model organism to study microbial dispersal and biogeography. METHODOLOGY/PRINCIPAL FINDINGS: The phylogeography of M. aeruginosa was studied based on a dataset of 311 rDNA internal transcribed spacer (ITS) sequences sampled from six continents. Richness of ITS sequences was high (239 ITS types were detected). Genetic divergence among ITS types averaged 4% (maximum pairwise divergence was 13%). Preliminary analyses revealed nearly completely unresolved phylogenetic relationships and a lack of genetic structure among all sequences due to extensive homoplasy at multiple hypervariable sites. After correcting for this, still no clear phylogeographic structure was detected, and no pattern of isolation by distance was found on a global scale. Concomitantly, genetic differentiation among continents was marginal, whereas variation within continents was high and was mostly shared with all other continents. Similarly, no genetic structure across climate zones was detected. CONCLUSIONS/SIGNIFICANCE: The high overall diversity and wide global distribution of common ITS types in combination with the lack of phylogeographic structure suggest that intercontinental dispersal of M. aeruginosa ITS types is not rare, and that this species might have a truly cosmopolitan distribution.

Limits to gene flow in a cosmopolitan marine planktonic diatom.

The role of geographic isolation in marine microbial speciation is hotly debated because of the high dispersal potential and large population sizes of planktonic microorganisms and the apparent lack of strong dispersal barriers in the open sea. Here, we show that gene flow between distant populations of the globally distributed, bloom-forming diatom species Pseudo-nitzschia pungens (clade I) is limited and follows a strong isolation by distance pattern. Furthermore, phylogenetic analysis implies that under appropriate geographic and environmental circumstances, like the pronounced climatic changes in the Pleistocene, population structuring may lead to speciation and hence may play an important role in diversification of marine planktonic microorganisms. A better understanding of the factors that control population structuring is thus essential to reveal the role of allopatric speciation in marine microorganisms.

Strong effects of amoebae grazing on the biomass and genetic structure of a Microcystis bloom (Cyanobacteria).

Despite its importance for bloom toxicity, the factors determining the population structure of cyanobacterial blooms are poorly understood. Here, we report the results of a two-year field survey of the population dynamics of Microcystis blooms in a small hypertrophic urban pond. Microscopic enumeration of Microcystis and its predators and parasites was combined with pigment and microcystin analysis and denaturing gradient gel electrophoresis of the ITS rDNA region to assess population dynamics and structure. Two main Microcystis morpho- and ITS types were revealed, corresponding to M. aeruginosa and M. viridis. In both years, high population densities of naked amoebae grazing on Microcystis coincided with rapid decreases in Microcystis biomass. In one year, there was a shift from heavily infested M. aeruginosa to the less-infested M. viridis, allowing the bloom to rapidly recover. The preference of amoebae for M. aeruginosa was confirmed by grazing experiments, in which several amoeba strains were capable of grazing down a strain of M. aeruginosa, but not of M. viridis. Zooplankton and chytrid parasites appeared to be of minor importance for these strong and fast reductions in Microcystis biomass. These findings demonstrate a strong impact of small protozoan grazers on the biomass and genetic structure of Microcystis blooms.

Natural hybrids in the marine diatom Pseudo-nitzschia pungens (Bacillariophyceae): genetic and morphological evidence.

Hybridization between genetically distinguishable taxa provides opportunities for investigating speciation. While hybridization is a common phenomenon in various macro-organisms, natural hybridization among micro-eukaryotes is barely studied. Here we used a nuclear and a chloroplast molecular marker and morphology to demonstrate the presence of natural hybrids between two genetically and morphologically distinct varieties of the marine planktonic diatom Pseudo-nitzschia pungens (vars. pungens and cingulata) in a contact zone in the northeast Pacific. Cloning and sequencing of the rDNA internal transcribed spacer region revealed strains containing ribotypes from both varieties, indicating hybridization. Both varieties were found to also have different chloroplast-encoded rbcL sequences. Hybrid strains were either hetero- or homoplastidial, as demonstrated by denaturing gradient gel electrophoresis, which is in accordance with expectations based on the mode of chloroplast inheritance in Pseudo-nitzschia. While most hybrids are probably first generation, there are also indications for further hybridization. Morphologically, the hybrids resembled var. pungens for most characters rather than having an intermediate morphology. Further research should focus on the hybridization frequency, by assessing the spatial and temporal extent of the contact zone, and hybrid fitness, to determine the amount of gene flow between the two varieties and its evolutionary consequences.