Einschätzen phylogenetischer Abgrenzungen von Taxa fademförmiger Xanthophyceae (Stramenopiles) mithilfe von DNA Sequenzanalysen und Morphologie

This thesis focuses on assessing taxonomic boundaries among the filamentous members of “yellow-green algae”, i.e. the class Xanthophyceae (Stramenopiles), which are widespread microalgae in freshwater or terrestrial habitats. The scarcity of morphological characters as well as their considerable plasticity requires the employment of molecular marker sequences to define monophyletic genera and species as well as for the unambiguous identification of new isolates. The genetic diversity and species boundaries within genera of Tribonemataceae, i.e Xanthonema Silva, Bumilleria Borzì, Bumilleriopsis Printz, Pseudobumilleriopsis Deason & Bold and Tribonema Derbès & Solier, were assessed using the highly variable psbA/rbcL spacer region as well as the more conserved full rbcL gene sequences. A method and a reference data base for the unambiguous identification of species of Heterococcus Chodat which uses DNA sequence variation in markers from plastid and nuclear genomes were established. In all studied cases the molecular-based species boundaries were more conservative than those defined by morphology. Within a single species almost identical genotypes were repeatedly recovered from strains of different geographic origins. However, the genotypes recovered from Antarctic strains were distinct from those of temperate regions.Die vorliegende Arbeit hat zum Ziel taxonomische Abgrenzungen bei fadenförmigen „Gelbgrünalgen“, d.h. Vertretern der Klasse Xanthophyceae (Stramenopiles), festzustellen. Fadenförmige Xanthophyceae sind weitverbreitete Mikroalgen im Süßwasser und in terrestrischen Lebensräumen. Da diese Algen arm an morphologischen Merkmalen sind und diese zudem eine beträchtliche Plastizität aufweisen, ist es notwendig, molekulare Markersequenzen einzusetzen, um monophyletische Gattungen und Arten zu definieren und neue Isolate auf Artniveau eindeutig bestimmen zu können. Die genetische Diversität und die Abgrenzungen von Gattungen der Tribonemataceae, d.h. Xanthonema Silva, Bumilleria Borzì, Bumilleriopsis Printz, Pseudobumilleriopsis Deason & Bold und Tribonema Derbès & Solier, wurden anhand hochvariabler Sequencen der psbA/rbcL Spacer-Region als auch konservierterer vollständigr rbcL Gensequenzen erfasst. Zur eindeutigen Identifizierung von Arten der Gattung Heterococcus Chodat wurden eine neue Methode und Referenzdaten erarbeitet, die auf Sequenzvariationen in Markermolekülen sowohl des Plastiden- als auch des Kerngenoms beruhen. In allen untersuchten Fällen waren die molekular-phylogenetisch begründeten Artabgrenzungen konservativer als die aufgrund morphologischer Merkmale. Innerhalb derselben Art wurden wiederholt nahezu identische Genotypen, die aber aus verschiedenen geographischen Ursprüngen stammten, aufgedeckt. Jedoch waren die Genotypen der Stämme, die aus der Antarktis isoliert wurden, verschieden von denen aus anderen temperaten geographischen Regionen

Congruence of chloroplast- and nuclear-encoded DNA sequence variations used to assess species boundaries in the soil microalga Heterococcus (Stramenopiles, Xanthophyceae).

BackgroundHeterococcus is a microalgal genus of Xanthophyceae (Stramenopiles) that is common and widespread in soils, especially from cold regions. Species are characterized by extensively branched filaments produced when grown on agarized culture medium. Despite the large number of species described exclusively using light microscopic morphology, the assessment of species diversity is hampered by extensive morphological plasticity.ResultsTwo independent types of molecular data, the chloroplast-encoded psbA/rbcL spacer complemented by rbcL gene and the internal transcribed spacer 2 of the nuclear rDNA cistron (ITS2), congruently recovered a robust phylogenetic structure. With ITS2 considerable sequence and secondary structure divergence existed among the eight species, but a combined sequence and secondary structure phylogenetic analysis confined to helix II of ITS2 corroborated relationships as inferred from the rbcL gene phylogeny. Intra-genomic divergence of ITS2 sequences was revealed in many strains. The 'monophyletic species concept', appropriate for microalgae without known sexual reproduction, revealed eight different species. Species boundaries established using the molecular-based monophyletic species concept were more conservative than the traditional morphological species concept. Within a species, almost identical chloroplast marker sequences (genotypes) were repeatedly recovered from strains of different origins. At least two species had widespread geographical distributions; however, within a given species, genotypes recovered from Antarctic strains were distinct from those in temperate habitats. Furthermore, the sequence diversity may correspond to adaptation to different types of habitats or climates.ConclusionsWe established a method and a reference data base for the unambiguous identification of species of the common soil microalgal genus Heterococcus which uses DNA sequence variation in markers from plastid and nuclear genomes. The molecular data were more reliable and more conservative than morphological data

Invading grass-like alga transforms rippled sand bars into bumpy muddy flats: arrival of a game changer in the Wadden Sea

In the wake of biological globalization, translocated species of high bio-engineering capacity increasingly change bottom topography of sedimentary coasts. A Vaucheria-taxon (Xanthophyceae) of unknown origin is spreading at the transition between intertidal and subtidal zones, while resident Vaucheria-species are confined to the upper shore in the Wadden Sea (European Atlantic). Near the island of Sylt, dense turfs of green filaments rapidly expanded over an area of 180 ha within 3 years. The unicellular filaments reach about 5 cm out of and 5 cm into the sediment. Felted rhizoids provide firm anchorage. Dry phytomass (up to 208 g m-2) was similar to that of intertidal seagrass beds. Residual filaments overwinter in the sediment and give rise to renewed growth in late spring. In addition, oospores germinate. Fine particles are trapped by the turf during summer, generating laminated cohesive mud. Muddy hummocks arise up to 20 cm above ambient sand flats, alternating with troughs but gradually merge into coherent and pertinacious plateaus of mud. This shift in bottom topography and sediment composition may potentially change the mud balance of tidal basins, and the capacity of tidal flats in catching up with accelerating sea-level rise

Unrecognized diversity and distribution of soil algae from Maritime Antarctica (Fildes Peninsula, King George Island)

Introduction: Eukaryotic algae in the top few centimeters of fellfield soils of ice-free Maritime Antarctica have many important effects on their habitat, such as being significant drivers of organic matter input into the soils and reducing the impact of wind erosion by soil aggregate formation. To better understand the diversity and distribution of Antarctic terrestrial algae, we performed a pilot study on the surface soils of Meseta, an ice-free plateau mountain crest of Fildes Peninsula, King George Island, being hardly influenced by the marine realm and anthropogenic disturbances. It is openly exposed to microbial colonization from outside Antarctica and connected to the much harsher and dryer ice-free zones of the continental Antarctic. A temperate reference site under mild land use, SchF, was included to further test for the Meseta algae distribution in a contrasting environment. Methods: We employed a paired-end metabarcoding analysis based on amplicons of the highly variable nuclear-encoded ITS2 rDNA region, complemented by a clone library approach. It targeted the four algal classes, Chlorophyceae, Trebouxiophyceae, Ulvophyceae, and Xanthophyceae, representing key groups of cold-adapted soil algae. Results: A surprisingly high diversity of 830 algal OTUs was revealed, assigned to 58 genera in the four targeted algal classes. Members of the green algal class Trebouxiophyceae predominated in the soil algae communities. The major part of the algal biodiversity, 86.1% of all algal OTUs, could not be identified at the species level due to insufficient representation in reference sequence databases. The classes Ulvophyceae and Xanthophyceae exhibited the most unknown species diversity. About 9% of the Meseta algae species diversity was shared with that of the temperate reference site in Germany. Discussion: In the small portion of algal OTUs for which their distribution could be assessed, the entire ITS2 sequence identity with references shows that the soil algae likely have a wide distribution beyond the Polar regions. They probably originated from soil algae propagule banks in far southern regions, transported by aeolian transport over long distances. The dynamics and severity of environmental conditions at the soil surface, determined by high wind currents, and the soil algae’s high adaptability to harsh environmental conditions may account for the high similarity of soil algal communities between the northern and southern parts of the Meseta

Development and investigation of the nanosensor-based apparatus to assess the psycho-emotional state of a person

Psycho-emotional state is one of the factors effecting human health. Its evaluation allows revealing hidden psychological trauma which can be reason of chronic stress, depression or psychosomatic disorders. Modern techniques of objective psycho-emotional state assessment involve a device which detects electrophysiological parameters of human body connected with emotional reaction and psychological condition. The present study covers development and testing of psycho-emotional state assessment device. The developed implement uses three methods of electrophysiological activity evaluation: electrocardiography, electroencephalography and galvanic skin response detection. The device represents hardware-software complex consisting of nanosensors, measuring unit, lead wires and laptop. Filters are excluded from the measuring circuit due to metrological parameters and noise immunity of implemented nanosensors. This solution minimizes signal distortion and allows measuring signals of 0.3 [mu]V and higher in a wide frequency range (0-10000 Hz) with minimal data loss. In addition, results of preliminary medical studies aimed to find correspondence between different psycho-emotional states and electrophysiological parameters are described. Impact of filters on electrophysiological studies was studied. According to the results conventional filters significantly distort EEG channel information. Further research will be directed to the creation of complete base of electrophysiological parameters related to a particular emotion

Unrecognized diversity and distribution of soil algae from Maritime Antarctica (Fildes Peninsula, King George Island)

IntroductionEukaryotic algae in the top few centimeters of fellfield soils of ice-free Maritime Antarctica have many important effects on their habitat, such as being significant drivers of organic matter input into the soils and reducing the impact of wind erosion by soil aggregate formation. To better understand the diversity and distribution of Antarctic terrestrial algae, we performed a pilot study on the surface soils of Meseta, an ice-free plateau mountain crest of Fildes Peninsula, King George Island, being hardly influenced by the marine realm and anthropogenic disturbances. It is openly exposed to microbial colonization from outside Antarctica and connected to the much harsher and dryer ice-free zones of the continental Antarctic. A temperate reference site under mild land use, SchF, was included to further test for the Meseta algae distribution in a contrasting environment.MethodsWe employed a paired-end metabarcoding analysis based on amplicons of the highly variable nuclear-encoded ITS2 rDNA region, complemented by a clone library approach. It targeted the four algal classes, Chlorophyceae, Trebouxiophyceae, Ulvophyceae, and Xanthophyceae, representing key groups of cold-adapted soil algae.ResultsA surprisingly high diversity of 830 algal OTUs was revealed, assigned to 58 genera in the four targeted algal classes. Members of the green algal class Trebouxiophyceae predominated in the soil algae communities. The major part of the algal biodiversity, 86.1% of all algal OTUs, could not be identified at the species level due to insufficient representation in reference sequence databases. The classes Ulvophyceae and Xanthophyceae exhibited the most unknown species diversity. About 9% of the Meseta algae species diversity was shared with that of the temperate reference site in Germany.DiscussionIn the small portion of algal OTUs for which their distribution could be assessed, the entire ITS2 sequence identity with references shows that the soil algae likely have a wide distribution beyond the Polar regions. They probably originated from soil algae propagule banks in far southern regions, transported by aeolian transport over long distances. The dynamics and severity of environmental conditions at the soil surface, determined by high wind currents, and the soil algae’s high adaptability to harsh environmental conditions may account for the high similarity of soil algal communities between the northern and southern parts of the Meseta

Repositories for Taxonomic Data: Where We Are and What is Missing

AbstractNatural history collections are leading successful large-scale projects of specimen digitization (images, metadata, DNA barcodes), thereby transforming taxonomy into a big data science. Yet, little effort has been directed towards safeguarding and subsequently mobilizing the considerable amount of original data generated during the process of naming 15,000–20,000 species every year. From the perspective of alpha-taxonomists, we provide a review of the properties and diversity of taxonomic data, assess their volume and use, and establish criteria for optimizing data repositories. We surveyed 4113 alpha-taxonomic studies in representative journals for 2002, 2010, and 2018, and found an increasing yet comparatively limited use of molecular data in species diagnosis and description. In 2018, of the 2661 papers published in specialized taxonomic journals, molecular data were widely used in mycology (94%), regularly in vertebrates (53%), but rarely in botany (15%) and entomology (10%). Images play an important role in taxonomic research on all taxa, with photographs used in &amp;gt;80% and drawings in 58% of the surveyed papers. The use of omics (high-throughput) approaches or 3D documentation is still rare. Improved archiving strategies for metabarcoding consensus reads, genome and transcriptome assemblies, and chemical and metabolomic data could help to mobilize the wealth of high-throughput data for alpha-taxonomy. Because long-term—ideally perpetual—data storage is of particular importance for taxonomy, energy footprint reduction via less storage-demanding formats is a priority if their information content suffices for the purpose of taxonomic studies. Whereas taxonomic assignments are quasifacts for most biological disciplines, they remain hypotheses pertaining to evolutionary relatedness of individuals for alpha-taxonomy. For this reason, an improved reuse of taxonomic data, including machine-learning-based species identification and delimitation pipelines, requires a cyberspecimen approach—linking data via unique specimen identifiers, and thereby making them findable, accessible, interoperable, and reusable for taxonomic research. This poses both qualitative challenges to adapt the existing infrastructure of data centers to a specimen-centered concept and quantitative challenges to host and connect an estimated $\le$2 million images produced per year by alpha-taxonomic studies, plus many millions of images from digitization campaigns. Of the 30,000–40,000 taxonomists globally, many are thought to be nonprofessionals, and capturing the data for online storage and reuse therefore requires low-complexity submission workflows and cost-free repository use. Expert taxonomists are the main stakeholders able to identify and formalize the needs of the discipline; their expertise is needed to implement the envisioned virtual collections of cyberspecimens. [Big data; cyberspecimen; new species; omics; repositories; specimen identifier; taxonomy; taxonomic data.]</jats:p

Species distinctions among closely related strains of Eustigmatophyceae (Stramenopiles) emphasizing ITS2 sequence-structure data: Eustigmatos and Vischeria

here is an increasing interest in the Eustigmatophyceae, a class of stramenopile microalgae, because they offer a variety of high-value health-beneficial compounds, e.g. polyunsaturated fatty acids (PUFAs), while concomitantly producing high biomass. Clarification of the taxonomy of these organisms at the species level is important in order to achieve reproducible results and constant yields of valuable compounds in their exploitation. Here the distinction of the, so far exclusively, morphologically defined species of the genera Eustigmatos and Vischeria was tested. Distinctions inferred from almost full 18S and ITS2 rRNA as well as plastid-encoded rbcL gene sequences were evaluated following a morphological investigation. The ITS2 secondary-structure-based phylogenies separated independent lineages (species) with long internal branches. This recommends ITS2 as a promising marker for a DNA metabarcoding approach (culture-independent biodiversity assessment). In contrast, the 18S V4 region which is commonly used in metabarcoding was almost invariant, whereas the almost full length sequences distinguished eight groups/types of strains. Monophyly of the species was supported by shared ITS2 secondary structure features, making them distinct from other eustigmatophyte lineages in concordance with phylogenetic analyses. No groups of strains were congruently supported by all three markers. Consequently, the previous distinction of two genera on the basis of morphology cannot be retained and the species should be accommodated in a single genus, Vischeria. Taxonomic changes among the species with the definition of epitypes, on the basis of cryopreserved strains, are recommended. Two findings point to a more complex evolutionary history of the species. The rbcL and nuclear markers resulted in disparate groupings of strains. In three species divergent intragenomic ITS2 paralogues were revealed. Therefore, a still broader taxon sampling, in conjunction with a deep sequencing approach, is needed for a more comprehensive understanding of the complex evolution of eustigmatophyte species

Congruence of chloroplast- and nuclear-encoded DNA sequence variations used to assess species boundaries in the soil microalga Heterococcus (Stramenopiles, Xanthophyceae).

BackgroundHeterococcus is a microalgal genus of Xanthophyceae (Stramenopiles) that is common and widespread in soils, especially from cold regions. Species are characterized by extensively branched filaments produced when grown on agarized culture medium. Despite the large number of species described exclusively using light microscopic morphology, the assessment of species diversity is hampered by extensive morphological plasticity.ResultsTwo independent types of molecular data, the chloroplast-encoded psbA/rbcL spacer complemented by rbcL gene and the internal transcribed spacer 2 of the nuclear rDNA cistron (ITS2), congruently recovered a robust phylogenetic structure. With ITS2 considerable sequence and secondary structure divergence existed among the eight species, but a combined sequence and secondary structure phylogenetic analysis confined to helix II of ITS2 corroborated relationships as inferred from the rbcL gene phylogeny. Intra-genomic divergence of ITS2 sequences was revealed in many strains. The 'monophyletic species concept', appropriate for microalgae without known sexual reproduction, revealed eight different species. Species boundaries established using the molecular-based monophyletic species concept were more conservative than the traditional morphological species concept. Within a species, almost identical chloroplast marker sequences (genotypes) were repeatedly recovered from strains of different origins. At least two species had widespread geographical distributions; however, within a given species, genotypes recovered from Antarctic strains were distinct from those in temperate habitats. Furthermore, the sequence diversity may correspond to adaptation to different types of habitats or climates.ConclusionsWe established a method and a reference data base for the unambiguous identification of species of the common soil microalgal genus Heterococcus which uses DNA sequence variation in markers from plastid and nuclear genomes. The molecular data were more reliable and more conservative than morphological data

Biotechnological Screening of Microalgal and Cyanobacterial Strains for Biogas Production and Antibacterial and Antifungal Effects

Microalgae and cyanobacteria represent a valuable natural resource for the generation of a large variety of chemical substances that are of interest for medical research, can be used as additives in cosmetics and food production, or as an energy source in biogas plants. The variety of potential agents and the use of microalgae and cyanobacteria biomass for the production of these substances are little investigated and not exploited for the market. Due to the enormous biodiversity of microalgae and cyanobacteria, they hold great promise for novel products. In this study, we investigated a large number of microalgal and cyanobacterial strains from the Culture Collection of Algae at Göttingen University (SAG) with regard to their biomass and biogas production, as well antibacterial and antifungal effects. Our results demonstrated that microalgae and cyanobacteria are able to generate a large number of economically-interesting substances in different quantities dependent on strain type. The distribution and quantity of some of these components were found to reflect phylogenetic relationships at the level of classes. In addition, between closely related species and even among multiple isolates of the same species, the productivity may be rather variable