Phylotranscriptomics reveals the reticulate evolutionary history of a widespread diatom species complex.

In contrast to surveys based on a few genes that often provide limited taxonomic resolution, transcriptomes provide a wealth of genomic loci that can resolve relationships among taxonomically challenging lineages. Diatoms are a diverse group of aquatic microalgae that includes important bioindicator species and many such lineages. One example is Nitzschia palea, a widespread species complex with several morphologically defined taxonomic varieties, some of which are critical pollution indicators. Morphological differences among the varieties are subtle and phylogenetic studies based on a few genes fail to resolve their evolutionary relationships. We conducted morphometric and transcriptome analyses of 10 Nitzschia palea strains to resolve the relationships among strains and taxonomic varieties. Nitzschia palea was resolved into three clades, one of which corresponds to a group of strains with narrow linear-lanceolate valves. The other morphological group recovered in the shape outline analysis was not monophyletic and consisted of two clades. Gene-tree concordance analyses and phylogenetic network estimations revealed patterns of incomplete lineage sorting and gene flow between intraspecific lineages. We detected reticulated evolutionary patterns among lineages with different morphologies, resulting in a putative recent hybrid. Our study shows that phylogenomic analyses of unlinked nuclear loci, complemented with morphometrics, can resolve complex evolutionary histories of recently diverged species complexes.

On Sea Turtle-associated Craspedostauros (Bacillariophyta), with Description of Three Novel Species.

The current study focuses on four species from the primarily marine diatom genus Craspedostauros that were observed growing attached to numerous sea turtles and sea turtle-associated barnacles from Croatia and South Africa. Three of the examined taxa, C. danayanus sp. nov., C. legouvelloanus sp. nov., and C. macewanii sp. nov., are described based on morphological and, whenever possible, molecular characteristics. The new taxa exhibit characters not previously observed in other members of the genus, such as the presence of more than two rows of cribrate areolae on the girdle bands, shallow perforated septa, and a complete reduction of the stauros. The fourth species, C. alatus, itself recently described from museum sea turtle specimens, is reported for the first time from loggerhead sea turtles rescued in Europe. A 3-gene phylogenetic analysis including DNA sequence data for three sea turtle-associated Craspedostauros species and other marine and epizoic diatom taxa indicated that Craspedostauros is monophyletic and sister to Achnanthes. This study, being based on a large number of samples and animal specimens analyzed and using different preservation and processing methods, provides new insights into the ecology and biogeography of the genus and sheds light on the level of intimacy and permanency in the host-epibiont interaction within the epizoic Craspedostauros species.

Continent-wide risk assessment for the establishment of nonindigenous species in Antarctica.

Invasive alien species are among the primary causes of biodiversity change globally, with the risks thereof broadly understood for most regions of the world. They are similarly thought to be among the most significant conservation threats to Antarctica, especially as climate change proceeds in the region. However, no comprehensive, continent-wide evaluation of the risks to Antarctica posed by such species has been undertaken. Here we do so by sampling, identifying, and mapping the vascular plant propagules carried by all categories of visitors to Antarctica during the International Polar Year's first season (2007-2008) and assessing propagule establishment likelihood based on their identity and origins and on spatial variation in Antarctica's climate. For an evaluation of the situation in 2100, we use modeled climates based on the Intergovernmental Panel on Climate Change's Special Report on Emissions Scenarios Scenario A1B [Nakicenovic N, Swart R, eds (2000) Special Report on Emissions Scenarios: A Special Report of Working Group III of the Intergovernmental Panel on Climate Change (Cambridge University Press, Cambridge, UK)]. Visitors carrying seeds average 9.5 seeds per person, although as vectors, scientists carry greater propagule loads than tourists. Annual tourist numbers (∼33,054) are higher than those of scientists (∼7,085), thus tempering these differences in propagule load. Alien species establishment is currently most likely for the Western Antarctic Peninsula. Recent founder populations of several alien species in this area corroborate these findings. With climate change, risks will grow in the Antarctic Peninsula, Ross Sea, and East Antarctic coastal regions. Our evidence-based assessment demonstrates which parts of Antarctica are at growing risk from alien species that may become invasive and provides the means to mitigate this threat now and into the future as the continent's climate changes.

Gomphonemavancampianum sp. nov. (Gomphonemataceae, Bacillariophyceae), a new large Gomphonema species from Europe.

During a French biomonitoring survey of the lakes in the region Rhône-Méditerranée-Corse, a large, unknown Gomphonema taxon was observed in a lake in the vicinity of the City of Lyon (Département du Rhône, France), that could not be identified using the currently available literature. Detailed light and scanning electron microscopy investigations revealed the presence of two types of valves, one bearing a thick marginal crest and valves lacking the crest. Following comparison with similar, mostly tropical Gomphonema species, the unknown taxon is described as new: Gomphonemavancampianum sp. nov. Discriminating features of the new species include the peculiar valve shape resembling a Chinese spoon, the broad upper valve part with acuminate tip, the presence of shallow depressions in the axial area, distinctly punctate striae and the occasional presence of the marginal crest. The new species was observed in several samples collected in an oligo- to mesotrophic, calcium-carbonate rich lake with a high ecological quality.

Five new species in the genus Staurosirella (Bacillariophyta) from European freshwater habitats.

Several populations belonging to the genus Staurosirella have been observed in European rivers that were previously identified as Staurosirellapinnata. In light of the recent taxonomic revisions of the genus Staurosirella, the morphology of the unknown Staurosirella populations has been critically investigated using light and scanning electron microscopy. Following the comparison with previously described Staurosirella species, five taxa could not be identified using the currently available literature on the genus. These five taxa are described as new based on differences in valve outline; shape, size and structure of the apical pore fields; structure of the striae; and the presence, position and structure of the marginal spines. Two new species were described using historic collection material: Staurosirellabinodiformis sp. nov. and Svanheurckiana sp. nov. Two new species were observed in samples from rivers in Flanders: S.marginostriata sp. nov. and S.stoksiana sp. nov. whereas a fifth species was observed in rivers from Iceland: S.jonssoniana sp. nov. All new species are compared with similar Staurosirella species worldwide. Notes are added on their ecological preferences derived from both physicochemical data and the associated diatom flora.

The influence of penguin activity on soil diatom assemblages on King George Island, Antarctica with the description of a new Luticola species.

Background: Ice-free areas in the Antarctic region are strongly limited. The presence of marine mammals and birds in those areas influence soil properties and vegetation composition. Studies on the terrestrial diatom flora in soils influenced by sea birds in the Maritime Antarctic region are scarce. Methods: Samples were collected from two transects on the western shore of the Admiralty Bay region. Light and scanning electron microscopic observations and statistical analyses were conducted to consider the impact of penguin rookeries on soil diatom assemblages. Results: The disturbance associated with the presence of penguin rookeries clearly influences the soil diatom diversity. Assemblages from areas with the highest nutrient input were characterized by a much lower diversity with only few species dominating the flora. One of recorded taxa could not be assigned to any of the known species. Therefore, based on the combination of morphological features analyzed using light and scanning electron microscopes and comparison with similar taxa in the Antarctic region and worldwide, the species is described hereby as new to science-Luticola kaweckae sp.nov. The new species is characteristic for soil habitats with strong penguin influence.

Two new Halamphora (Bacillariophyta) species from the marine coasts off Livingston Island, Antarctica.

During a survey of the marine benthic diatom flora on the coasts off Livingston Island (South Shetland Islands, Maritime Antarctic Region), two Halamphora species that could not be identified based on the currently available literature, were observed. Detailed light and scanning electron microscopy observations and thorough comparison with similar taxa in the literature revealed that both taxa should be described as new species. The first taxon, Halamphorakenderoviana sp. nov., was most likely misidentified in past Antarctic studies, and included within the range of another taxon, Halamphoracoffeaeformis. Analysis of literature data showed that the second new taxon, Halamphoramoncheviana sp. nov., has been previously reported from the Antarctic Continent (but as an unidentified species). The new taxa are compared with similar Halamphora taxa worldwide. Data on their ecology and distribution are also provided.

Cultivating epizoic diatoms provides insights into the evolution and ecology of both epibionts and hosts.

Our understanding of the importance of microbiomes on large aquatic animals-such as whales, sea turtles and manatees-has advanced considerably in recent years. The latest observations indicate that epibiotic diatom communities constitute diverse, polyphyletic, and compositionally stable assemblages that include both putatively obligate epizoic and generalist species. Here, we outline a successful approach to culture putatively obligate epizoic diatoms without their hosts. That some taxa can be cultured independently from their epizoic habitat raises several questions about the nature of the interaction between these animals and their epibionts. This insight allows us to propose further applications and research avenues in this growing area of study. Analyzing the DNA sequences of these cultured strains, we found that several unique diatom taxa have evolved independently to occupy epibiotic habitats. We created a library of reference sequence data for use in metabarcoding surveys of sea turtle and manatee microbiomes that will further facilitate the use of environmental DNA for studying host specificity in epizoic diatoms and the utility of diatoms as indicators of host ecology and health. We encourage the interdisciplinary community working with marine megafauna to consider including diatom sampling and diatom analysis into their routine practices.

A time-calibrated multi-gene phylogeny of the diatom genus Pinnularia.

Pinnularia is an ecologically important and species-rich genus of freshwater diatoms (Bacillariophyceae) showing considerable variation in frustule morphology. Interspecific evolutionary relationships were inferred for 36 Pinnularia taxa using a five-locus dataset. A range of fossil taxa, including newly discovered Middle Eocene forms of Pinnularia, was used to calibrate a relaxed molecular clock analysis and investigate temporal aspects of the genus' diversification. The multi-gene approach resulted in a well-resolved phylogeny of three major clades and several subclades that were frequently, but not universally, delimited by valve morphology. The genus Caloneis was not recovered as monophyletic, confirming that, as currently delimited, this genus is not evolutionarily meaningful and should be merged with Pinnularia. The Pinnularia-Caloneis complex is estimated to have diverged between the Upper Cretaceous and the early Eocene, implying a ghost range of at least 10 million year (Ma) in the fossil record.

Diatom Genus Hyalosira (Rhabdonematales emend.) and Resolution of its Polyphyly in Grammatophoraceae and Rhabdonemataceae with a New Genus, Placosira, and Five New Hyalosira Species.

Hyalosira gene sequences are divided into two clades within different families. We examined authentic material of Hyalosira (isotype material of H. obtusangula, synonymous with H. delicatula) and voucher specimens of published sequences, and pooled our observations of Hyalosira-like taxa from benthic and epizoic habitats in several parts of the globe. The two molecular clades corresponded to two morphological groups, with Hyalosira obtusangula associated with Grammatophoraceae. We emend the description and provide lectotypification for Hyalosira and propose Placosira to encompass the taxa in the other clade, associated with Rhabdonemataceae. We propose that Hyalosira has uniseriate to triseriate striae, sometimes different on valve face and mantle. Copulae in most species had shallow septa, though in one they were moderately deep. All species had girdle bands bearing two rows of areolae separated by a midrib. We name five new species of Hyalosira. Morphologies of taxa in the Placosira clade were superficially similar to Hyalosira but differed in having areolae with ricae, a single row of areolae on the girdle bands, and tubular rimoportulae on the valve-face-mantle junction. Hyalosira hustedtiana Patrick should revert to its original position in Striatella until the appropriate genus can be determined. We emend Rhabdonematales to encompass Rhabdonemataceae, Grammatophoraceae and Tabellariaceae.

Extinction of austral diatoms in response to large-scale climate dynamics in Antarctica.

Despite evidence for microbial endemism, an understanding of the impact of geological and paleoclimate events on the evolution of regional protist communities remains elusive. Here, we provide insights into the biogeographical history of Antarctic freshwater diatoms, using lacustrine fossils from mid-Miocene and Quaternary Antarctica, and dovetail this dataset with a global inventory of modern freshwater diatom communities. We reveal the existence of a diverse mid-Miocene diatom flora bearing similarities with several former Gondwanan landmasses. Miocene cooling and Plio-Pleistocene glaciations triggered multiple extinction waves, resulting in the selective depauperation of this flora. Although extinction dominated, in situ speciation and new colonizations ultimately shaped the species-poor, yet highly adapted and largely endemic, modern Antarctic diatom flora. Our results provide a more holistic view on the scale of biodiversity turnover in Neogene and Pleistocene Antarctica than the fragmentary perspective offered by macrofossils and underscore the sensitivity of lacustrine microbiota to large-scale climate perturbations.

Geographical variation in the diatom communities associated with loggerhead sea turtles (Caretta caretta).

Epizoic diatoms form an important part of micro-epibiota of marine vertebrates such as whales and sea turtles. The present study explores and compares the diversity and biogeography of diatom communities growing on the skin and shell of loggerhead sea turtles (Caretta caretta) from four different localities: Adriatic Sea (Croatia), Ionian Sea (Greece), South Africa and Florida Bay (USA) using both light and scanning electron microscopy. We observed almost 400 diatom taxa belonging to more than 100 genera. Diatom communities from Greece and Croatia showed the highest similarity and were statistically different from those recorded from South Africa and Florida. Part of this variation could be attributed to differences in sampling techniques; however, we believe that geography had an important role. In general, contrary to several previous observations from sea turtles, the presumably exclusively epizoic diatoms contributed less than common benthic taxa to the total diatom flora, which might have been related to the loggerhead feeding behavior. Moreover, skin samples differed from carapace samples in having a distinct diatom composition with a higher proportion of the putative true epizoonts. Our results indicate that epizoic diatom communities differ according to loggerhead geographical location and substrate (skin vs. carapace). The relative abundances of common benthic diatoms and putative exclusive epizoic taxa may inform about sea turtle habitat use or behavior though detailed comparisons among different host species have yet to be performed.

Latitudinal Biogeographic Structuring in the Globally Distributed Moss Ceratodon purpureus.

Biogeographic patterns of globally widespread species are expected to reflect regional structure, as well as connectivity caused by occasional long-distance dispersal. We assessed the level and drivers of population structure, connectivity, and timescales of population isolation in one of the most widespread and ruderal plants in the world - the common moss Ceratodon purpureus. We applied phylogenetic, population genetic, and molecular dating analyses to a global (n = 147) sampling data set, using three chloroplast loci and one nuclear locus. The plastid data revealed several distinct and geographically structured lineages, with connectivity patterns associated with worldwide, latitudinal "bands." These imply that connectivity is strongly influenced by global atmospheric circulation patterns, with dispersal and establishment beyond these latitudinal bands less common. Biogeographic patterns were less clear within the nuclear marker, with gene duplication likely hindering the detection of these. Divergence time analyses indicated that the current matrilineal population structure in C. purpureus has developed over the past six million years, with lineages diverging during the late Miocene, Pliocene, and Quaternary. Several colonization events in the Antarctic were apparent, as well as one old and distinct Antarctic clade, possibly isolated on the continent since the Pliocene. As C. purpureus is considered a model organism, the matrilineal biogeographic structure identified here provides a useful framework for future genetic and developmental studies on bryophytes. Our general findings may also be relevant to understanding global environmental influences on the biogeography of other organisms with microscopic propagules (e.g., spores) dispersed by wind.

Global radiation in a rare biosphere soil diatom.

Soil micro-organisms drive the global carbon and nutrient cycles that underlie essential ecosystem functions. Yet, we are only beginning to grasp the drivers of terrestrial microbial diversity and biogeography, which presents a substantial barrier to understanding community dynamics and ecosystem functioning. This is especially true for soil protists, which despite their functional significance have received comparatively less interest than their bacterial counterparts. Here, we investigate the diversification of Pinnularia borealis, a rare biosphere soil diatom species complex, using a global sampling of >800 strains. We document unprecedented high levels of species-diversity, reflecting a global radiation since the Eocene/Oligocene global cooling. Our analyses suggest diversification was largely driven by colonization of novel geographic areas and subsequent evolution in isolation. These results illuminate our understanding of how protist diversity, biogeographical patterns, and members of the rare biosphere are generated, and suggest allopatric speciation to be a powerful mechanism for diversification of micro-organisms.

Extensive Cryptic Diversity in the Terrestrial Diatom Pinnularia borealis (Bacillariophyceae).

With the increasing application of molecular techniques for diatom species discovery and identification, it is important both from a taxonomic as well as an ecological and applied perspective, to understand in which groups morphological species delimitation is congruent with molecular approaches, or needs reconsideration. Moreover, such studies can improve our understanding of morphological trait evolution in this important group of microalgae. In this study, we used morphometric analysis on light microscopy (LM) micrographs in SHERPA, detailed scanning electron microscopy (SEM), and cytological observations in LM to examine 70 clones belonging to eight distinct molecular lineages of the cosmopolitan terrestrial diatom Pinnularia borealis. Due to high within-lineage variation, no conclusive morphological separation in LM nor SEM could be detected. Morphological stasis due to the "low-morphology" problem or stabilizing selection, as well as parallel/convergent evolution, phenotypic plasticity and structural inheritance are discussed as potential drivers for the observations. Altogether, P. borealis is truly cryptic, in contrast to the majority of other diatom species complexes which turned out to be pseudo-cryptic following detailed morphological analysis.

Hippos (Hippopotamus amphibius): The animal silicon pump.

While the importance of grasslands in terrestrial silicon (Si) cycling and fluxes to rivers is established, the influence of large grazers has not been considered. Here, we show that hippopotamuses are key actors in the savannah biogeochemical Si cycle. Through a detailed analysis of Si concentrations and stable isotope compositions in multiple ecosystem compartments of a savannah-river continuum, we constrain the processes influencing the Si flux. Hippos transport 0.4 metric tons of Si day-1 by foraging grass on land and directly egesting in the water. As such, they bypass complex retention processes in secondary soil Si pools. By balancing internal processes of dissolution and precipitation in the river sediment, we calculate that hippos affect up to 76% of the total Si flux. This can have a large impact on downstream lake ecosystems, where Si availability directly affects primary production in the diatom-dominated phytoplankton communities.

Responses of aquatic organisms to metal pollution in a lowland river in Flanders: a comparison of diatoms and macroinvertebrates.

The role of macroinvertebrates and diatoms as indicator for metal pollution was investigated by assessing both biota along a metal gradient in the Belgian river the Dommel. Macroinvertebrates and diatoms were sampled in summer and winter and physical-chemical characteristics of the water were measured at four different sample periods and related to sediment characteristics. Although metal concentrations, except cadmium, in the water nowhere exceeded water quality standards, high metal concentrations were measured in the sediment, indicating historical contamination of the Dommel. At the sites that were situated downstream of the pollution source, high levels of conductivity and chloride were measured in the water. Redundancy Analysis (RDA) indicated pH, phosphate and zinc as the significant environmental variables explaining each respectively 7.7%, 11.6% and 22.6% of the macroinvertebrate community composition. Two clusters could be separated, with Gammarus pulex, Leptocerus interruptus, Baetis rhodani and Cloeon dipterum associated with low zinc concentrations and Tubificidae, Asellus aquaticus, Erpobdella sp. and Chironomus thummi-plumosus associated with higher zinc concentrations. Ammonium (10.6%), conductivity (16.5%), chloride (11.4%) and zinc (5.9%) turned out to be significant variables explaining the diatom community structure. Based on physical-chemical differences and species composition, three different groups could be separated. With this Tabellaria flocculosa and Fragilaria capucina var. rumpens were associated with low metal concentrations, Gomphonema parvulum and Nitzschia palea with elevated concentrations and Eolimna minima and Sellaphora seminulum with high zinc concentrations. In conclusion, the diatom community best reflected the metal gradient. With regard to water quality indices, those based on macroinvertebrates best followed the metal pollution gradient and were most strongly correlated with physical-chemical variables of water and sediment. This study indicated that to assess the effect of metal pollution in lowland rivers, the combined use of macroinvertebrates and diatoms is more appropriate than the use of both biota separately.

Historical processes constrain patterns in global diatom diversity.

There is a long-standing belief that microbial organisms have unlimited dispersal capabilities, are therefore ubiquitous, and show weak or absent latitudinal diversity gradients. In contrast, using a global freshwater diatom data set, we show that latitudinal gradients in local and regional genus richness are present and highly asymmetric between both hemispheres. Patterns in regional richness are explained by the degree of isolation of lake districts, while the number of locally coexisting diatom genera is highly constrained by the size of the regional diatom pool, habitat availability, and the connectivity between habitats within lake districts. At regional to global scales, historical factors explain significantly more of the observed geographic patterns in genus richness than do contemporary environmental conditions. Together, these results stress the importance of dispersal and migration in structuring diatom communities at regional to global scales. Our results are consistent with predictions from the theory of island biogeography and metacommunity concepts and likely underlie the strong provinciality and endemism observed in the relatively isolated diatom floras in the Southern Hemisphere.

Molecular evidence for distinct Antarctic lineages in the cosmopolitan terrestrial diatoms Pinnularia borealis and Hantzschia amphioxys.

Recent morphology-based studies indicate that freshwater diatom floras in the Antarctic comprise a significant share of endemics among a majority of apparently cosmopolitan species. Given the widespread (pseudo)cryptic species diversity in diatoms, we assessed the molecular divergence and temperature-dependent growth characteristics between Antarctic and non-Antarctic strains for two presumed species with a cosmopolitan distribution, namely Pinnularia borealis and Hantzschia amphioxys. Molecular phylogenies based on the plastid gene rbcL and the nuclear 28S rDNA (D1-D3 region) revealed that both taxa consist of multiple lineages, each including a distinct Antarctic lineage. A molecular clock estimates the origin of P. borealis at 35.8 (30-47) million years (Ma) ago, making this the oldest known diatom species complex. The Antarctic P. borealis lineage is estimated to have diverged 7.8 (2-15) Ma ago, after the geographical and thermal isolation of the Antarctic continent. Despite not being psychrophilic, the Antarctic lineages of P. borealis and H. amphioxys have a lower optimal growth temperature and upper lethal temperature than most lineages from more temperate regions, indicating niche differentiation. Together, this suggests that many presumed cosmopolitan Antarctic diatom species are in fact species complexes, possibly containing Antarctic endemics with low temperature preferences.

Variable temperature effects of Open Top Chambers at polar and alpine sites explained by irradiance and snow depth.

Environmental manipulation studies are integral to determining biological consequences of climate warming. Open Top Chambers (OTCs) have been widely used to assess summer warming effects on terrestrial biota, with their effects during other seasons normally being given less attention even though chambers are often deployed year-round. In addition, their effects on temperature extremes and freeze-thaw events are poorly documented. To provide robust documentation of the microclimatic influences of OTCs throughout the year, we analysed temperature data from 20 studies distributed across polar and alpine regions. The effects of OTCs on mean temperature showed a large range (-0.9 to 2.1 °C) throughout the year, but did not differ significantly between studies. Increases in mean monthly and diurnal temperature were strongly related (R(2)  = 0.70) with irradiance, indicating that PAR can be used to predict the mean warming effect of OTCs. Deeper snow trapped in OTCs also induced higher temperatures at soil/vegetation level. OTC-induced changes in the frequency of freeze-thaw events included an increase in autumn and decreases in spring and summer. Frequency of high-temperature events in OTCs increased in spring, summer and autumn compared with non-manipulated control plots. Frequency of low-temperature events was reduced by deeper snow accumulation and higher mean temperatures. The strong interactions identified between aspects of ambient environmental conditions and effects of OTCs suggest that a detailed knowledge of snow depth, temperature and irradiance levels enables us to predict how OTCs will modify the microclimate at a particular site and season. Such predictive power allows a better mechanistic understanding of observed biotic response to experimental warming studies and for more informed design of future experiments. However, a need remains to quantify OTC effects on water availability and wind speed (affecting, for example, drying rates and water stress) in combination with microclimate measurements at organism level.