TY - THES A1 - Montulet, Orianne T1 - Functional characterization of putative interactors of the Cellulose Synthase Complex Y1 - 2024 ER - TY - THES A1 - Apodiakou, Anastasia T1 - Analysis of the regulation of SDI genes, unravelling the role of the SLIM1 transcription factor, and the SNRK3.15 kinase in Arabidopsis under sulfur deprivation Y1 - 2024 ER - TY - THES A1 - Seerangan, Kumar T1 - Actin-based regulation of cell and tissue scale morphologenesis in develpoping leaves Y1 - 2023 ER - TY - THES A1 - Hagemann, Justus T1 - On the molecular evolution of sengis (Macroscelidea) N2 - This thesis focuses on the molecular evolution of Macroscelidea, commonly referred to as sengis. Sengis are a mammalian order belonging to the Afrotherians, one of the four major clades of placental mammals. Sengis currently consist of twenty extant species, all of which are endemic to the African continent. They can be separated in two families, the soft-furred sengis (Macroscelididae) and the giant sengis (Rhynchocyonidae). While giant sengis can be exclusively found in forest habitats, the different soft-furred sengi species dwell in a broad range of habitats, from tropical rain-forests to rocky deserts. Our knowledge on the evolutionary history of sengis is largely incomplete. The high level of superficial morphological resemblance among different sengi species (especially the soft-furred sengis) has for example led to misinterpretations of phylogenetic relationships, based on morphological characters. With the rise of DNA based taxonomic inferences, multiple new genera were defined and new species described. Yet, no full taxon molecular phylogeny exists, hampering the answering of basic taxonomic questions. This lack of knowledge can be to some extent attributed to the limited availability of fresh-tissue samples for DNA extraction. The broad African distribution, partly in political unstable regions and low population densities complicate contemporary sampling approaches. Furthermore, the DNA information available usually covers only short stretches of the mitochondrial genome and thus a single genetic locus with limited informational content. Developments in DNA extraction and library protocols nowadays offer the opportunity to access DNA from museum specimens, collected over the past centuries and stored in natural history museums throughout the world. Thus, the difficulties in fresh-sample acquisition for molecular biological studies can be overcome by the application of museomics, the research field which emerged from those laboratory developments. This thesis uses fresh-tissue samples as well as a vast collection museum specimens to investigate multiple aspects about the macroscelidean evolutionary history. Chapter 4 of this thesis focuses on the phylogenetic relationships of all currently known sengi species. By accessing DNA information from museum specimens in combination of fresh tissue samples and publicly available genetic resources it produces the first full taxon molecular phylogeny of sengis. It confirms the monophyly of the genus Elephantulus and discovers multiple deeply divergent lineages within different species, highlighting the need for species specific approaches. The study furthermore focuses on the evolutionary time frame of sengis by evaluating the impact of commonly varied parameters on tree dating. The results of the study show, that the mitochondrial information used in previous studies to temporal calibrate the Macroscelidean phylogeny led to an overestimation of node ages within sengis. Especially soft-furred sengis are thus much younger than previously assumed. The refined knowledge of nodes ages within sengis offer the opportunity to link e.g. speciation events to environmental changes. Chapter 5 focuses on the genus Petrodromus with its single representative Petrodromus tetradactylus. It again exploits the opportunities of museomics and gathers a comprehensive, multi-locus genetic dataset of P. tetradactylus individuals, distributed across most the known range of this species. It reveals multiple deeply divergent lineages within Petrodromus, whereby some could possibly be associated to previously described sub-species, at least one was formerly unknown. It underscores the necessity for a revision of the genus Petrodromus through the integration of both molecular and morphological evidence. The study, furthermore identifies changing forest distributions through climatic oscillations as main factor shaping the genetic structure of Petrodromus. Chapter 6 uses fresh tissue samples to extent the genomic resources of sengis by thirteen new nuclear genomes, of which two were de-novo assembled. An extensive dataset of more than 8000 protein coding one-to-one orthologs allows to further refine and confirm the temporal time frame of sengi evolution found in Chapter 4. This study moreover investigates the role of gene-flow and incomplete lineage sorting (ILS) in sengi evolution. In addition it identifies clade specific genes of possible outstanding evolutionary importance and links them to potential phenotypic traits affected. A closer investigation of olfactory receptor proteins reveals clade specific differences. A comparison of the demographic past of sengis to other small African mammals does not reveal a sengi specific pattern. N2 - Diese Dissertation untersucht die molekulare Evolution von Macroscelidea, auch als Sengis oder Rüsselspringer bezeichnet. Sengis sind eine Ordnung der Afrotheria, einer der vier Hauptkladen der plazentalen Säugetiere. Aktuell gibt es zwanzig beschriebene Sengiarten, die alle ausschließlich auf dem afrikanischen Kontinent vorkommen. Sengis können in zwei Familien unterteilt werden: die Elephantenspitzmäuse zusammen mit den Rüsselratten bilden die Macroscelididae und die Rüsselhündchen die Rhynchocyonidae. Während Rhynchocyonidae ausschließlich in Waldhabitaten zu finden sind, bewohnen verschiedene Macroscelididaearten ein breites Spektrum von Lebensräumen, von tropischen Regenwäldern bis zu felsigen Wüsten. Unser Wissen über die evolutionäre Geschichte der Sengis ist äußerst unvollständig. Der hohe Grad an morphologischer Ähnlichkeit zwischen verschiedenen Sengiarten (insbesondere innerhalb der Macroscelididae) hat beispielsweise zu Fehlinterpretationen phylogenetischer Beziehungen auf der Grundlage morphologischer Merkmale geführt. Mit dem Aufkommen DNA-basierter taxonomischer Forschung wurden mehrere neue Gattungen definiert und neue Arten beschrieben. Dennoch existiert derzeit keine vollständige molekulare Phylogenie, was die Beantwortung grundlegender taxonomischer Fragen und tiefergehende evolutionsbiologische Analysen erschwert. Dieser Mangel an Wissen kann zum Teil auf die begrenzte Verfügbarkeit von frischen Gewebeproben für die DNA-Extraktion zurückgeführt werden. Die weite Verbreitung in Afrika, teilweise in politisch instabilen Regionen und geringe Populationssdichten von Sengis erschweren das Sammeln von frischem Probenmaterial, was für die Extraktion von DNA genutzt werden kann. Darüber hinaus deckt die bis jetzt verfügbare DNA-Information über Sengis häufig nur kurze Abschnitte des mitochondrialen Genoms ab und damit einen einzelnen genetischen Lokus mit begrenztem Informationsgehalt. Fortentwicklungen von DNA-Extraktions-Protokollen und Library-Protokollen bieten heutzutage die Möglichkeit, auf DNA von Museumsexemplaren zuzugreifen, die über die letzten Jahrhunderte gesammelt und in Naturkundemuseen weltweit aufbewahrt werden. Somit können die Schwierigkeiten bei der Beschaffung von Frischproben für molekularbiologische Studien überwunden werden. Diese Dissertation verwendet sowohl Frischgewebeproben als auch eine umfangreiche Sammlung von Museumssproben, um verschiedene Aspekte der evolutionären Geschichte der Sengis molekularbiologisch zu untersuchen. Kapitel 4 dieser Dissertation konzentriert sich auf die phylogenetischen Beziehungen aller derzeit bekannten Sengiarten. Durch das Generieren von DNA-Information aus Museumsexemplaren in Kombination mit Frischgewebeproben und öffentlich verfügbaren genetischen Ressourcen wird die erste vollständige molekulare Phylogenie aller Rüsselspringer erzeugt. Die Studie bestätigt die Monophylie der Gattung Elephantulus und entdeckt mehrere tief divergente Linien innerhalb verschiedener Arten, was die Notwendigkeit speziesbezogener Ansätze verdeutlicht. Die Studie konzentriert sich außerdem auf den Zeitrahmen der Sengi-Evolution, indem sie die Auswirkungen häufig variierter Parameter auf die Datierung von Stammbäumen untersucht. Die Ergebnisse zeigen, dass die mitochondriale Information, die in früheren Studien zur zeitlichen Kalibrierung der Macroscelidean-Phylogenie verwendet wurde, zu einer Überschätzung des Alters von Arttrennungen innerhalb der Rüsselspringer geführt hat. Insbesondere die Macroscelididae sind daher viel jünger als zuvor angenommen. Das präzisere Wissen über das evolutionäre Alter von Rüsselspringern bietet die Möglichkeit, beispielsweise Artaufspaltungen mit Umweltveränderungen zu verknüpfen. Kapitel 5 konzentriert sich auf die Gattung Petrodromus mit ihrem einzigen Vertreter Petrodromus tetradactylus. Es nutzt erneut die Museomics und sammelt einen umfassenden, genetischen Datensatz von P. tetradactylus-Individuen, die über den größten Teil des bekannten Verbreitungsgebiets dieser Art verteilt sind. Es zeigt mehrere tief divergente Linien innerhalb von Petrodromus auf, wobei einige mit zuvor beschriebenen Unterarten in Verbindung gebracht werden könnten, mindestens eine aber zuvor unbekannt war. Die Ergebnisse verdeutlichen die Notwendigkeit einer taxonomischen Überarbeitung der Gattung Petrodromus durch das Zusammenführen sowohl molekularer als auch morphologischer Indizien. Die Studie identifizier außerdem sich ändernde Waldverteilungen durch klimatische Schwankungen als Hauptfaktor, der die genetische Struktur von Petrodromus formt. Kapitel 6 verwendet Frischgewebeproben, um die genomischen Ressourcen der Rüsselspringer durch dreizehn neue nukleare Genome zu erweitern, von denen zwei de-novo assembliert wurden. Ein umfangreicher Datensatz von mehr als 8000 protein-kodierenden 1:1-Orthologen ermöglicht es, den zeitlichen Rahmen der Rüsselspringerevolution, der in Kapitel 4 gefunden wurde, weiter zu verfeinern und zu bestätigen. Diese Studie untersucht außerdem die Rolle von Genfluss auf die Evolution der Rüsselspringer. Darüber hinaus identifiziert sie für bestimmte Kladen spezifische Gene von möglicherweise herausragender evolutionärer Bedeutung und verknüpft diese mit potenziell betroffenen phänotypischen Merkmalen. Eine genauere Untersuchung von Geruchsrezeptorproteinen zeigt kladespezifische Unterschiede auf. KW - sengis KW - evolution KW - molecular dating KW - biogeography KW - comparative genomics KW - Biogeographie KW - vergleichende Genomik KW - Evolution KW - molekulare Datierung KW - Sengis Y1 - 2024 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:kobv:517-opus4-641975 ER - TY - JOUR A1 - Córdoba, Sandra Correa A1 - Tong, Hao A1 - Burgos, Asdrubal A1 - Zhu, Feng A1 - Alseekh, Saleh A1 - Fernie, Alisdair R. A1 - Nikoloski, Zoran T1 - Identification of gene function based on models capturing natural variability of Arabidopsis thaliana lipid metabolism JF - Nature Communications N2 - The use of automated tools to reconstruct lipid metabolic pathways is not warranted in plants. Here, the authors construct Plant Lipid Module for Arabidopsis rosette using constraint-based modeling, demonstrate its integration in other plant metabolic models, and use it to dissect the genetic architecture of lipid metabolism. Lipids play fundamental roles in regulating agronomically important traits. Advances in plant lipid metabolism have until recently largely been based on reductionist approaches, although modulation of its components can have system-wide effects. However, existing models of plant lipid metabolism provide lumped representations, hindering detailed study of component modulation. Here, we present the Plant Lipid Module (PLM) which provides a mechanistic description of lipid metabolism in the Arabidopsis thaliana rosette. We demonstrate that the PLM can be readily integrated in models of A. thaliana Col-0 metabolism, yielding accurate predictions (83%) of single lethal knock-outs and 75% concordance between measured transcript and predicted flux changes under extended darkness. Genome-wide associations with fluxes obtained by integrating the PLM in diel condition- and accession-specific models identify up to 65 candidate genes modulating A. thaliana lipid metabolism. Using mutant lines, we validate up to 40% of the candidates, paving the way for identification of metabolic gene function based on models capturing natural variability in metabolism. KW - Biochemical networks KW - Biochemical reaction networks KW - Genetic models KW - Plant molecular biology Y1 - 2023 U6 - https://doi.org/10.1038/s41467-023-40644-9 SN - 2041-1723 VL - 14 IS - 1 PB - Springer Nature CY - London ER - TY - JOUR A1 - Cheng, Feng A1 - Dennis, Alice B. A1 - Osuoha, Josephine Ijeoma A1 - Canitz, Julia A1 - Kirschbaum, Frank A1 - Tiedemann, Ralph T1 - A new genome assembly of an African weakly electric fish (Campylomormyrus compressirostris, Mormyridae) indicates rapid gene family evolution in Osteoglossomorpha JF - BMC genomics N2 - Background Teleost fishes comprise more than half of the vertebrate species. Within teleosts, most phylogenies consider the split between Osteoglossomorpha and Euteleosteomorpha/Otomorpha as basal, preceded only by the derivation of the most primitive group of teleosts, the Elopomorpha. While Osteoglossomorpha are generally species poor, the taxon contains the African weakly electric fish (Mormyroidei), which have radiated into numerous species. Within the mormyrids, the genus Campylomormyrus is mostly endemic to the Congo Basin. Campylomormyrus serves as a model to understand mechanisms of adaptive radiation and ecological speciation, especially with regard to its highly diverse species-specific electric organ discharges (EOD). Currently, there are few well-annotated genomes available for electric fish in general and mormyrids in particular. Our study aims at producing a high-quality genome assembly and to use this to examine genome evolution in relation to other teleosts. This will facilitate further understanding of the evolution of the osteoglossomorpha fish in general and of electric fish in particular. Results A high-quality weakly electric fish (C. compressirostris) genome was produced from a single individual with a genome size of 862 Mb, consisting of 1,497 contigs with an N50 of 1,399 kb and a GC-content of 43.69%. Gene predictions identified 34,492 protein-coding genes, which is a higher number than in the two other available Osteoglossomorpha genomes of Paramormyrops kingsleyae and Scleropages formosus. A Computational Analysis of gene Family Evolution (CAFE5) comparing 33 teleost fish genomes suggests an overall faster gene family turnover rate in Osteoglossomorpha than in Otomorpha and Euteleosteomorpha. Moreover, the ratios of expanded/contracted gene family numbers in Osteoglossomorpha are significantly higher than in the other two taxa, except for species that had undergone an additional genome duplication (Cyprinus carpio and Oncorhynchus mykiss). As potassium channel proteins are hypothesized to play a key role in EOD diversity among species, we put a special focus on them, and manually curated 16 Kv1 genes. We identified a tandem duplication in the KCNA7a gene in the genome of C. compressirostris. Conclusions We present the fourth genome of an electric fish and the third well-annotated genome for Osteoglossomorpha, enabling us to compare gene family evolution among major teleost lineages. Osteoglossomorpha appear to exhibit rapid gene family evolution, with more gene family expansions than contractions. The curated Kv1 gene family showed seven gene clusters, which is more than in other analyzed fish genomes outside Osteoglossomorpha. The KCNA7a, encoding for a potassium channel central for EOD production and modulation, is tandemly duplicated which may related to the diverse EOD observed among Campylomormyrus species. KW - Campylomormyrus KW - Pacbio sequencing KW - Gene family KW - Osteoglossomorpha KW - Kv1 Y1 - 2023 U6 - https://doi.org/10.1186/s12864-023-09196-6 SN - 1471-2164 VL - 24 IS - 1 PB - BMC CY - London ER - TY - JOUR A1 - Peter, Lena A1 - Wendering, Désirée Jacqueline A1 - Schlickeiser, Stephan A1 - Hoffmann, Henrike A1 - Noster, Rebecca A1 - Wagner, Dimitrios Laurin A1 - Zarrinrad, Ghazaleh A1 - Münch, Sandra A1 - Picht, Samira A1 - Schulenberg, Sarah A1 - Moradian, Hanieh A1 - Mashreghi, Mir-Farzin A1 - Klein, Oliver A1 - Gossen, Manfred A1 - Roch, Toralf A1 - Babel, Nina A1 - Reinke, Petra A1 - Volk, Hans-Dieter A1 - Amini, Leila A1 - Schmueck-Henneresse, Michael T1 - Tacrolimus-resistant SARS-CoV-2-specific T cell products to prevent and treat severe COVID-19 in immunosuppressed patients JF - Molecular therapy methods and clinical development N2 - Solid organ transplant (SOT) recipients receive therapeutic immunosuppression that compromises their immune response to infections and vaccines. For this reason, SOT patients have a high risk of developing severe coronavirus disease 2019 (COVID-19) and an increased risk of death from severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection. Moreover, the efficiency of immunotherapies and vaccines is reduced due to the constant immunosuppression in this patient group. Here, we propose adoptive transfer of SARS-CoV-2-specific T cells made resistant to a common immunosuppressant, tacrolimus, for optimized performance in the immunosuppressed patient. Using a ribonucleoprotein approach of CRISPR-Cas9 technology, we have generated tacrolimus-resistant SARS-CoV-2-specific T cell products from convalescent donors and demonstrate their specificity and function through characterizations at the single-cell level, including flow cytometry, single-cell RNA (scRNA) Cellular Indexing of Transcriptomes and Epitopes (CITE), and T cell receptor (TCR) sequencing analyses. Based on the promising results, we aim for clinical validation of this approach in transplant recipients. Additionally, we propose a combinatory approach with tacrolimus, to prevent an overshooting immune response manifested as bystander T cell activation in the setting of severe COVID-19 immunopathology, and tacrolimus-resistant SARS-CoV-2-specific T cell products, allowing for efficient clearance of viral infection. Our strategy has the potential to prevent severe COVID-19 courses in SOT or autoimmunity settings and to prevent immunopathology while providing viral clearance in severe non-transplant COVID-19 cases. Y1 - 2022 U6 - https://doi.org/10.1016/j.omtm.2022.02.012 SN - 2329-0501 VL - 25 SP - 52 EP - 73 PB - Cell Press CY - Cambridge ER - TY - JOUR A1 - Tomowski, Maxi A1 - Lozada-Gobilard, Sissi Donna A1 - Jeltsch, Florian A1 - Tiedemann, Ralph T1 - Recruitment and migration patterns reveal a key role for seed banks in the meta-population dynamics of an aquatic plant JF - Scientific reports N2 - Progressive habitat fragmentation threatens plant species with narrow habitat requirements. While local environmental conditions define population growth rates and recruitment success at the patch level, dispersal is critical for population viability at the landscape scale. Identifying the dynamics of plant meta-populations is often confounded by the uncertainty about soil-stored population compartments. We combined a landscape-scale assessment of an amphibious plant's population structure with measurements of dispersal complexity in time to track dispersal and putative shifts in functional connectivity. Using 13 microsatellite markers, we analyzed the genetic structure of extant Oenanthe aquatica populations and their soil seed banks in a kettle hole system to uncover hidden connectivity among populations in time and space. Considerable spatial genetic structure and isolation-by-distance suggest limited gene flow between sites. Spatial isolation and patch size showed minor effects on genetic diversity. Genetic similarity found among extant populations and their seed banks suggests increased local recruitment, despite some evidence of migration and recent colonization. Results indicate stepping-stone dispersal across adjacent populations. Among permanent and ephemeral demes the resulting meta-population demography could be determined by source-sink dynamics. Overall, these spatiotemporal connectivity patterns support mainland-island dynamics in our system, highlighting the importance of persistent seed banks as enduring sources of genetic diversity. Y1 - 2023 U6 - https://doi.org/10.1038/s41598-023-37974-5 SN - 2045-2322 VL - 13 IS - 1 PB - Springer Nature CY - London ER - TY - JOUR A1 - Arend, Marius A1 - Zimmer, David A1 - Xu, Rudan A1 - Sommer, Frederik A1 - Mühlhaus, Timo A1 - Nikoloski, Zoran T1 - Proteomics and constraint-based modelling reveal enzyme kinetic properties of Chlamydomonas reinhardtii on a genome scale JF - Nature Communications N2 - Metabolic engineering of microalgae offers a promising solution for sustainable biofuel production, and rational design of engineering strategies can be improved by employing metabolic models that integrate enzyme turnover numbers. However, the coverage of turnover numbers for Chlamydomonas reinhardtii, a model eukaryotic microalga accessible to metabolic engineering, is 17-fold smaller compared to the heterotrophic cell factory Saccharomyces cerevisiae. Here we generate quantitative protein abundance data of Chlamydomonas covering 2337 to 3708 proteins in various growth conditions to estimate in vivo maximum apparent turnover numbers. Using constrained-based modeling we provide proxies for in vivo turnover numbers of 568 reactions, representing a 10-fold increase over the in vitro data for Chlamydomonas. Integration of the in vivo estimates instead of in vitro values in a metabolic model of Chlamydomonas improved the accuracy of enzyme usage predictions. Our results help in extending the knowledge on uncharacterized enzymes and improve biotechnological applications of Chlamydomonas. KW - Computational models KW - Enzymes KW - Proteomics Y1 - 2023 U6 - https://doi.org/10.1038/s41467-023-40498-1 SN - 2041-1723 VL - 14 IS - 1 PB - Springer Nature CY - London ER - TY - JOUR A1 - Ferreira, Clara Mendes A1 - Dammhahn, Melanie A1 - Eccard, Jana T1 - So many choices, so little time BT - food preference and movement vary with the landscape of fear JF - Ecology and evolution N2 - Spatial and temporal variation in perceived predation risk is an important determinant of movement and foraging activity of animals. Foraging in this landscape of fear, individuals need to decide where and when to move, and what resources to choose. Foraging theory predicts the outcome of these decisions based on energetic trade-offs, but complex interactions between perceived predation risk and preferences of foragers for certain functional traits of their resources are rarely considered. Here, we studied the interactive effects of perceived predation risk on food trait preferences and foraging behavior in bank voles (Myodes glareolus) in experimental landscapes. Individuals (n = 19) were subjected for periods of 24 h to two extreme, risk-uniform landscapes (either risky or safe), containing 25 discrete food patches, filled with seeds of four plant species in even amounts. Seeds varied in functional traits: size, nutrients, and shape. We evaluated whether and how risk modifies forager preference for functional traits. We also investigated whether perceived risk and distance from shelter affected giving-up density (GUD), time in patches, and number of patch visits. In safe landscapes, individuals increased time spent in patches, lowered GUD and visited distant patches more often compared to risky landscapes. Individuals preferred bigger seeds independent of risk, but in the safe treatment they preferred fat-rich over carb-rich seeds. Thus, higher densities of resource levels remained in risky landscapes, while in safe landscapes resource density was lower and less diverse due to selective foraging. Our results suggest that the interaction of perceived risk and dietary preference adds an additional layer to the cascading effects of a landscape of fear which affects biodiversity at resource level. KW - foraging behavior KW - functional traits KW - giving-up density KW - myodes glareolus KW - perceived predation risk KW - seed ecology Y1 - 2023 U6 - https://doi.org/10.1002/ece3.10330 SN - 2045-7758 VL - 13 IS - 7 PB - Wiley CY - Hoboken ER -