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The salinity gradient separating marine and freshwater environments is a major ecological divide, and the mechanisms by which most organisms adapt to new salinity environments are poorly understood. Diatoms are a lineage of ancestrally marine microalgae that have repeatedly colonized and diversified in freshwaters. Cyclotella cryptica is a euryhaline diatom that naturally tolerates a broad range of salinities, thus providing a powerful system for understanding the genomic mechanisms for mitigating and acclimating to low salinity. To understand how diatoms mitigate acute hypoosmotic stress, we abruptly shifted C. cryptica from seawater to freshwater and performed transcriptional profiling during the first 10 hours. Freshwater shock dramatically remodeled the transcriptome, with ~50% of the genome differentially expressed in at least one time point. The peak response occurred within 1 hour, with strong repression of genes involved in cell growth and osmolyte production, and strong induction of specific stress defense genes. Transcripts largely returned to baseline levels within 4–10 hours, with growth resuming shortly thereafter, suggesting that gene expression dynamics may be useful for predicting acclimation. Moreover, comparison to a transcriptomics study of C. cryptica following months-long acclimation to freshwater revealed little overlap between the genes and processes differentially expressed in cells exposed to acute stress versus fully acclimated conditions. Altogether, this study highlights the power of time-resolved transcriptomics to reveal fundamental insights into how cells dynamically respond to an acute environmental shift and provides new insights into how diatoms mitigate natural salinity fluctuations and have successfully diversified across freshwater habitats worldwide.

Despite the obstacles facing marine colonists, most lineages of aquatic organisms have colonized and diversified in freshwaters repeatedly. These transitions can trigger rapid morphological or physiological change and, on longer timescales, lead to increased rates of speciation and extinction. Diatoms are a lineage of ancestrally marine microalgae that have diversified throughout freshwater habitats worldwide. We generated a phylogenomic dataset of genomes and transcriptomes for 59 diatom taxa to resolve freshwater transitions in one lineage, the Thalassiosirales. Although most parts of the species tree were consistently resolved with strong support, we had difficulties resolving a Paleocene radiation, which affected the placement of one freshwater lineage. This and other parts of the tree were characterized by high levels of gene tree discordance caused by incomplete lineage sorting and low phylogenetic signal. Despite differences in species trees inferred from concatenation versus summary methods and codons versus amino acids, traditional methods of ancestral state reconstruction supported six transitions into freshwaters, two of which led to subsequent species diversification. Evidence from gene trees, protein alignments, and diatom life history together suggest that habitat transitions were largely the product of homoplasy rather than hemiplasy, a condition where transitions occur on branches in gene trees not shared with the species tree. Nevertheless, we identified a small set of putatively hemiplasious genes, many of which have been associated with shifts to low salinity, indicating that hemiplasy played a small but potentially important role in freshwater adaptation. Accounting for differences in evolutionary outcomes, in which some taxa became locked into freshwaters while others were able to return to the ocean or become salinity generalists, might help further distinguish different sources of adaptive mutation in freshwater diatoms.

Many diatom groups are known for widespread (pseudo)cryptic species diversity and Pinnularia gibba group is one of them. Recently, Kollar et al. (2019) delimited species within the group by means of a polyphasic approach, providing an evidence for the existence of fifteen species. In order to further guide the systematic revision of the group, the present study focuses on the evolution and morphology of the P. gibba group. Using representatives of the fifteen species we used fossils to constrain and calibrate a multi-gene species-level phylogeny. Although many species are morphologically highly similar, significant differences in cell size were detected in different sections of the tree, suggestive of ongoing morphological differentiation. We further used the phylogenetic analysis to assess marker resolution for DNA barcoding, showing that the proposed diatom barcode markers rbcL and V4 SSU rDNA can distinguish closely related (pseudo)cryptic species of the group.

The salinity gradient separating marine and freshwater environments represents a major ecological divide for microbiota, yet the mechanisms by which marine microbes have adapted to and ultimately diversified in freshwater environments are poorly understood. Here, we take advantage of a natural evolutionary experiment: the colonization of the brackish Baltic Sea by the ancestrally marine diatom Skeletonema marinoi. To understand how diatoms respond to low salinity, we characterized transcriptomic responses of S. marinoi grown in a common garden. Our experiment included eight genotypes from source populations spanning the Baltic Sea salinity cline. Changes in gene expression revealed a shared response to salinity across genotypes, where low salinities induced profound changes in cellular metabolism, including upregulation of carbon fixation and storage compound biosynthesis, and increased nutrient demand and oxidative stress. Nevertheless, the genotype effect overshadowed the salinity effect, as genotypes differed significantly in their response, both in the magnitude and direction of gene expression. Intraspecific differences included regulation of transcription and translation, nitrogen metabolism, cell signaling, and aerobic respiration. The high degree of intraspecific variation in gene expression observed here highlights an important but often overlooked source of biological variation associated with how diatoms respond and adapt to environmental change.

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.

Terrestrial biota in the Antarctic are more globally distinct and highly structured biogeographically than previously believed, but information on biogeographic patterns and endemism in freshwater communities is largely lacking. We studied biogeographic patterns of Antarctic freshwater diatoms based on the analysis of species occurrences in a dataset of 439 lakes spread across the Antarctic realm. Highly distinct diatom floras, both in terms of composition and richness, characterize Continental Antarctica, Maritime Antarctica and the sub-Antarctic islands, with marked biogeographic provincialism in each region. A total of 44% of all species is estimated to be endemic to the Antarctic, and most of them are confined to a single biogeographic region. The level of endemism significantly increases with increasing latitude and geographic isolation. Our results have implications for conservation planning, and suggest that successful dispersal of freshwater diatoms to and within the Antarctic is limited, fostering the evolution of highly endemic diatom floras.

The 11th Symposium on Polar Science/Ordinary sessions: [OB] Polar Biology Wed. 2 Dec.

The increasing urbanization process is hypothesized to drastically alter (semi-)natural environments with a concomitant major decline in species abundance and diversity. Yet, studies on this effect of urbanization, and the spatial scale at which it acts, are at present inconclusive due to the large heterogeneity in taxonomic groups and spatial scales at which this relationship has been investigated among studies. Comprehensive studies analysing this relationship across multiple animal groups and at multiple spatial scales are rare, hampering the assessment of how biodiversity generally responds to urbanization. We studied aquatic (cladocerans), limno-terrestrial (bdelloid rotifers) and terrestrial (butterflies, ground beetles, ground- and web spiders, macro-moths, orthopterans and snails) invertebrate groups using a hierarchical spatial design, wherein three local-scale (200 m x 200 m) urbanization levels were repeatedly sampled across three landscape-scale (3 km x 3 km) urbanization levels. We tested for local and landscape urbanization effects on abundance and species richness of each group, whereby total richness was partitioned into the average richness of local communities and the richness due to variation among local communities. Abundances of the terrestrial active dispersers declined in response to local urbanization, with reductions up to 85% for butterflies, while passive dispersers did not show any clear trend. Species richness also declined with increasing levels of urbanization, but responses were highly heterogeneous among the different groups with respect to the richness component and the spatial scale at which urbanization impacts richness. Depending on the group, species richness declined due to biotic homogenization and/or local species loss. This resulted in an overall decrease in total richness across groups in urban areas. These results provide strong support to the general negative impact of urbanization on abundance and species richness within habitat patches and highlight the importance of considering multiple spatial scales and taxa to assess the impacts of urbanization on biodiversity. Belgian Science Policy Office, Grant/Award Number: P07/4; Conselho Nacional de Desenvolvimento Cientifico e Tecnologico, Grant/Award Number: 45968/2012-1 Piano, E (reprint author), Univ Turin, Dept Life Sci & Syst Biol, Turin, Italy. elena.piano@unito.it

Recent morphology-based investigations of freshwater Arctic diatoms suggest that many species remain to be discovered, reflecting a unique polar flora. During a survey of the freshwater diatom flora of northern Billefjorden, including the Petuniabukta fjord region (Spitsbergen), several morphodemes belonging to the Achnanthidium minutissimum species complex were recorded. Molecular phylogenies based on rbcL, 28S and 18S sequences, including single cells from Canada and strains from Marion Island (sub-Antarctica) and GenBank revealed the presence of 12 distinct A. minutissimum complex lineages, of which three contained strains from Spitsbergen. One Arctic lineage is described as a new species. Achnanthidium digitatum sp. nov. is morphologically characterized by narrow, linear to only slightly lanceolate valves and usually two areolae per stria. The two remaining Arctic lineages are in need of a more complete morphological and molecular comparison with other representatives of the A. minutissimum complex to clarify their taxonomic identity. It is argued that implementation of molecular data in the taxonomy of Achnanthidium will be essential to solve the taxonomic problems associated with this group, eventually resulting in a better understanding of the biogeography and niche differentiation of different species belonging to the A. minutissimum complex. Ideally, this should be based on more variable genes than the currently widely used 18S, which does not have a species level resolution in the A. minutissimum complex.

A recent detailed survey of the Maritime Antarctic diatom flora using a fine-grained taxonomy resulted in the description of many new species of Pinnularia in general and the section Distantes, including the P. borealis species complex, in particular. Moreover, DNA-based studies of P. borealis revealed that many more species need to be described within this complex. During a survey of the freshwater littoral diatom flora of James Ross Island (Ulu Peninsula) and Vega Island in Maritime Antarctica, a previously unknown chain-forming species in the P. borealis species complex of section Distantes was cultured from three different localities. Molecular phylogenies based on the nuclear-encoded D1-D3 large-subunit ribosomal DNA and plastid rbcL genes revealed that all cultures belong to a distinct highly supported lineage within the P. borealis species complex. Pinnularia catenaborealis sp. nov. is characterised by the presence of small spines located on a raised, thin silica ridge that almost entirely surrounds the valve face near the valve face/mantle junction, and the presence of small silica plates near the apices. In culture, P. catenaborealis formed chains of several tens of cells and in oxidised natural material, chains up to seven frustules were observed. Pinnularia catenaborealis is described from the littoral zone of freshwater Black Lake (Ulu Peninsula, James Ross Island) and has also been observed on nearby Vega Island. Although P. borealis is generally regarded as a (semi-) terrestrial diatom complex mainly occurring in (moist) soils and mosses, P. catenaborealis was found in freshwater habitats with an alkaline pH and low conductivity.

Although polar regions are characterised by extreme environment conditions with overall low temperatures and often pronounced diurnal to seasonal temperature fluctuations, diatoms (Bacillariophyceae) thrive in a wide range of nonmarine polar habitats, suggesting that they are well adapted to these harsh conditions. In this study, tolerance to freezing of vegetative and resting cells of 17 polar and nine temperate diatom strains, belonging to different morphospecies and originating from both freshwater and terrestrial habitats, was experimentally tested. Most strains were newly isolated from sites in Maritime Antarctica (James Ross Island and Vega Island), the High Arctic (Spitsbergen) and Europe. Cultures were exposed to five freezing treatments differing in temperature (-4, -20, -40 and -180 degrees C), freezing rate (gradual, abrupt), time (1 h, 12 h) and thawing rate (slow, fast). Results indicated that diatoms were sensitive to experimental freezing. Freezing temperatures had a significant effect on strain survival: all strains survived -4 degrees C; most survived -20 degrees C; five survived -40 and four of these (all belonging to the Pinnularia borealis complex) survived freezing in liquid nitrogen (-180 degrees C). The cooling and thawing rate had a significant impact on survival: abrupt cooling and slow thawing resulted in much lower survival rates than gradual cooling and fast thawing. Resting cells showed better growth than vegetative cells after freezing treatment but only in the -4 degrees C treatment. Surprisingly, no striking differences in growth recovery were observed between polar and temperate strains.

Despite the frequent application of diatoms in palaeoecological research in the High Arctic, our knowledge concerning the diversity, ecology, community associations, community dynamics and survival strategies of High Arctic freshwater littoral diatom floras is still rather limited. In the present study, the diversity, ecological characteristics and community structures of the freshwater littoral diatom flora from ponds and lakes of Petuniabukta (Spitsbergen, Svalbard Archipelago) were studied. In total, 87 samples from the littoral zone of 53 freshwater ponds and lakes were examined using light and scanning electron microscopy. A highly diverse diatom flora dominated by Achnanthidium and Nitzschia was observed. In total, 310 diatom taxa belonging to 59 genera were found, of which about one-third could not be identified beyond the genus level. Based on species composition, four main diatom communities were distinguished. Differences in environmental characteristics, such as habitat type, habitat diversity, habitat stability, streams, water pH and influence of the fjord, biota and human activities, appeared to be important factors in determining the observed diatom communities. The high number of unidentified taxa in this study indicates that a profound revision of the Arctic diatom flora is highly desired, not only to improve our fundamental knowledge concerning the diversity, ecology, community associations and biogeography of Arctic diatoms but also to aid applied (palaeo)environmental sciences.

Diatoms are one of the most abundant and arguably the most species-rich group of protists. Diatom species delimitation has often been based exclusively on the recognition of morphological discontinuities without investigation of other lines of evidence. Even though DNA sequences and reproductive experiments have revealed several examples of (pseudo)cryptic diversity, our understanding of diatom species boundaries and diversity remains limited. The cosmopolitan pennate raphid diatom genus Pinnularia represents one of the most taxon-rich diatom genera. In this study, we focused on the delimitation of species in one of the major clades of the genus, the Pinnularia subgibba group, based on 105 strains from a worldwide origin. We compared genetic distances between the sequences of seven molecular markers and selected the most variable pair, the mitochondrial cox1 and nuclear encoded LSU rDNA, to formulate a primary hypothesis on the species limits using three single-locus automated species delimitation methods. We compared the DNA-based primary hypotheses with morphology and with other available lines of evidence. The results indicate that our data set comprised 15 species of the P. subgibba group. The vast majority of these taxa have an uncertain taxonomic identity, suggesting that several may be unknown to science and/or members of (pseudo)cryptic species complexes within the P. subgibba group.

Incomplete reproductive isolation between genetically distinct taxa provides an interesting opportunity for speciation and adaptation studies. This phenomenon is well-described in macro-organisms, but less experimental evidence is available for unicellular eukaryotes. Here, we document the sympatric occurrence of genetically differentiated populations of the pennate model diatom Seminavis robusta in coastal subtidal biofilm communities and show widespread potential for gene flow between them. Based on sequence variation in the plastid-encoded rbcL gene, three distinct clades were identified. Morphological variation between the clades reflected their phylogenetic relationships, with subtle differences in valve morphology in the most distant clade compared to the other two clades, which were indistinguishable. Using a large number of experimental crosses we showed that, although reproductive output was significantly lower compared to the majority of within-clade crosses, approximately 34.5% of the inter-clade crosses resulted in viable and fertile progeny. While the nature of the incomplete reproductive isolation remains unknown, its occurrence in natural diatom populations represents an additional mechanism contributing to population genetic structuring and adaptation and can spur further research into the mechanisms of species divergence and the maintenance of species identity in the presence of gene flow.

Diatoms constitute the most diverse group of microalgae and have long been recognised for their large biotechnological potential. In the wake of growing research interest in new model species and development of commercial applications, there is a pressing need for long-term preservation of diatom strains. While cryopreservation using dimethylsulfoxide (DMSO) as a cryoprotective agent is the preferred method for long-term strain preservation, many diatom species cannot be successfully cryopreserved using DMSO. Therefore, in this study, we studied cryopreservation success in six different diatom species, representing the major morphological and ecological diatom groups, using a range of DMSO concentrations and Plant Vitrification Solution 2 (PVS2) as an alternative cryoprotectant to DMSO. In addition, we tested whether suppressing bacterial growth by antibiotics accelerates the post-thaw recovery process. Our results show that the effects of cryoprotectant choice, its concentration and the addition of antibiotics are highly species specific. In addition, we showed that PVS2 and antibiotics are useful agents to optimize cryopreservation of algae that cannot survive the traditional cryopreservation protocol using DMSO. We conclude that a species-specific approach will remain necessary to develop protocols for diatom cryopreservation and to increase their representation in public culture collections.

Cryoconite holes are small, extreme habitats, widespread in the ablation zones of gla-ciers worldwide. They can provide a suitable environment for microorganisms including bacteria, cyanobacteria, algae, fungi, and invertebrates. Diatoms have been previously recovered from cryoconite holes of Greenland and of Svalbard, and recent findings from Antarctica suggest that cryoconite holes may harbor a unique diatom flora distinct from other aquatic habitats nearby. In the present study, we characterize the diatom communi-ties of Nordenskiöld glacier cryoconite holes in Billefjorden (Svalbard, Spitsbergen), and multivariate approaches were used to compare them with three freshwater localities in the immediate vicinity to investigate possible sources of the species pool. We found cryoconite holes to have similar or greater average genus-richness than adjacent lake/ ponds habitats, even though lower numbers of valves were recovered. Overall, cryoconite hole diatom communities differed significantly from those observed in lakes, suggesting that other sources actively contribute to these communities than nearby lakes alone. This further suggests that (i) diatoms present in cryoconite might not exclusively originate from aquatic habitats, but also from (semi-) terrestrial ones; and (ii) that a much wider area than the immediate surroundings should be considered as a possible source for cryoconite diatom flora.

During a survey conducted on the freshwater diatom flora of small shallow pools on the Ecology Glacier forefield (King George Island, Maritime Antarctic Region), an unknown spine-bearing chain-forming Pinnularia species, belonging to the Pinnularia borealis species complex, was found. Although it closely resembles the recently described Pinnularia catenaborealis from James Ross Island and Vega Island (Antarctic Peninsula), a unique set of morphological characteristics revealed in both light and scanning electron microscopy clearly discriminates the specimens of King George Island as a new species. Pinnularia subcatenaborealis Kochman-Kędziora, Pinseel & Van de Vijver sp. nov. can be distinguished from P. catenaborealis by an overall smaller valve size, the presence of irregularly formed silica outgrowths on the mantle and small, irregular plates located near the apices. The new taxon is so far only recorded from a small pool with circumneutral pH and very low conductivity.

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., It is generally thought many microbes, owing to their ubiquity and dispersal capability, lack biogeographic structuring and clear speciation patterns compared to macroorganisms. However, Pinseel et al. demonstrate multiple cycles of colonization and diversification in Pinnularia borealis, a rare biosphere soil diatom.

Diatoms constitute an important and diverse component of terrestrial protist communities but remain poorly studied, especially in the Antarctic realm. Here, we investigated the diversity and community structure of the terrestrial diatom flora from the Ulu Peninsula, James Ross Island (Maritime Antarctic Region) using a morphology-based dataset and physico-chemical measurements. A total of 97 taxa belonging to 27 genera was identified in 59 samples from terrestrial environments, including soils and rock walls. The flora was dominated by the genera Hantzschia, Luticola, and Humidophila. Eight distinct diatom assemblages could be distinguished and were mainly structured by differences in environmental characteristics such as vegetation coverage, moisture, conductivity, pH, and nutrient concentrations. In general, James Ross Island harboured a unique diatom flora as evidenced by very low similarity values with other (sub-)Antarctic localities. Only 16% of the taxa have a typical cosmopolitan distribution, whereas 70% showed a restricted Antarctic distribution, supporting previous indications of high species-level endemism in environments characterized by harsh abiotic conditions. In addition, several of the cosmopolitan species uncovered in this study might harbour substantial levels of hidden diversity, including endemic taxa, as previously revealed for the Pinnularia borealis species complex on James Ross Island. Taken together, the present study improves our knowledge and understanding of the diversity, ecology, and community structure of the terrestrial diatom flora of Ulu Peninsula and highlights that soils and wet rock walls represent important terrestrial habitats in this transitional zone between Maritime and Continental Antarctica.

During a survey of freshwater diatoms from lakes in the region of Petuniabukta on Spitsbergen (Svalbard Archipelago) a new Gomphonema species, G. svalbardense sp. nov., has been recorded. The new taxon was previously cited in the literature as G. angustatum var. undulatum but this identification proved to be erroneous. Detailed morphology description of G. svalbardense based on light and scanning electron microscopy is presented in this paper and the morphological features of the taxon have been compared with similar species. Gomphonema svalbardense is characterized by its typical linear, almost naviculoid outline with undulating margins, with clearly inflated central part, asymmetric central area, lateral raphe with simple straight proximal endings and the weakly radiate striae. History, ecology and biogeography of the species, mainly based on literature data, have been included.

During a survey of the freshwater littoral diatom flora from lakes and ponds in the region of Petuniabukta on Spitsbergen (Svalbard Archipelago, High Arctic region), a new Achnanthidium species, A . petuniabuktianum sp. nov., has been recorded. Achnanthidium petuniabuktianum is a new representative of the A . pyrenaicum group as evidenced by its curved distal raphe fissures. The present paper describes in detail the morphology of A . petuniabuktianum based on light and scanning electron microscopy. The new taxon is characterized in having linear valves with parallel margins and broadly rounded, never protracted apices and a characteristic—in light microscopy hardly discernible—striation pattern consisting of very short striae composed of one or two small, always slit-like areolae. Based on current results, Achnanthidium petuniabuktianum is a benthic taxon occurring in epilithic and epiphytic habitats in the littoral zones of freshwater lakes and ponds.