Le phylum des chaetognathes constitue une énigme zoologique très ancienne, puisque ses caractéristiques morphologiques et développementales contradictoires ne permettent pas de le classer aisément au sein des animaux bilatériens. Pour tenter de résoudre ce problème, nous avons séquencé une collection d’ESTs pour l’espèce Spadella cephaloptera et développé une approche phylogénomique en utilisant les protéines ribosomiques comme marqueurs et une stratégie originale de taxons composites. Nous avons trouvé que les chaetognathes occupaient une position de groupe frère de tous les autres protostomiens, une position originale qui échappe à la ‘nouvelle vision’ de la phylogénie des animaux et possède des implications pour l’orientation des caractères embryologiques au sein des métazoaires. Nous avons confirmé cette position phylogénétique en nous focalisant sur les gènes Hox des chaetognathes. En outre, nous avons tenté de résoudre l’organisation génomique des gènes Hox de chaetognathes et nous avons rassemblé des éléments qui suggèrent une distribution dispersée des gènes Hox dans le génome. Dans un second temps, une analyse exhaustive des données génomiques accumulées a mis en évidence de nouvelles caractéristiques génomiques inattendues chez les chaetognathes. Nous avons tout d’abord identifié une large duplication génomique caractérisée par un taux élevé de rétention des gènes dupliqués. Nous avons également détecté la présence de trans-splicing pour une fraction importante de transcrits, qui s’accompagne d’une transcription en opéron. Enfin, nous avons découvert un polymorphisme génomique de grande ampleur aux niveaux nucléotidiques et structuraux au sein de la population de référence de Sormiou, dont nous avons démontré qu’il n’était pas lié à un phénomène de spéciation cryptique. Nous avons réalisé une description détaillée de ces variations génétiques dans la région du gène Hox1, où elles sont induites par l’insertion polymorphe d’éléments génétiques mobiles. D’un autre côté, nous avons étudié la diversité génétique mitochondriale dans la population et nous avons identifié plusieurs lignées distinctes, séparées par une divergence moléculaire très élevée et par des remaniements structuraux. L’origine de ce polymorphisme et son impact sur la physiologie et la régulation génique constituent des pistes de recherche prometteuses. Globalement, ce travail a montré qu’au delà de son intérêt comme organisme modèle pour comprendre l’évolution des bilatériens, S. cephaloptera pourrait permettre d’étudier l’impact du polymorphisme génomique sur l’évolution. / The chaetognath phylum represents a longstanding zoological conundrum, as its contrasting morphological and developmental characters do not allow a convincing assignment among bilaterian lineages. To tackle this issue, we sequenced ESTs for Spadella cephaloptera and set up a phylogenomic approach based on a ribosomal protein dataset and an original composite taxon strategy. We found that chaetognaths are most likely branched as a protostome sister-group, which challenges ‘new view’ of animal phylogeny and provides insights into the evolution of developmental processes among metazoans. We brought further evidence for this phylogenetic position by focusing on chaetognath Hox genes. We attempted to resolve the genomic organization of the 13 chaetognath genes and we found clues of a dispersed organization. Then, a careful examination of the bulk of genomic data gathered has pointed out new unexpected unusual genomic features of chaetognaths. We identified an extensive gene duplication followed by a high retention of duplicated genes. We also determined that a large part of S. cephaloptera transcripts underwent trans-splicing associated with operonic transcription. Strikingly, we finally uncovered a tremendous genomic polymorphism at both nucleotide and structural levels within the reference population of Sormiou, and showed it is not caused by cryptic speciation. We provided a detailed account of genomic structural variations in the region surrounding Hox1 gene, which have been mediated by polymorphic insertion of mobile genetic elements. Alternatively, we explored the mitochondrial genetic diversity in the population and recovered several divergent mitochondrial lineages, split by phylum-level molecular divergence and structural rearrangments. The origin of this polymorphism as well as its impact on the physiology and genic regulation are challenging questions to think about. As a whole, this work stresses the interest of S. cephaloptera as a model organism, not only to study the evolution of bilaterian body plans, but also the impact of genomic polymorphism on organismal evolution.

No description available.

<p> The higher taxonomic groups within Prokaryotes are presently distinguished mainly on the basis of their branching in phylogenetic trees. In most cases, no molecular, biochemical or physiological characteristics are known that are uniquely shared by species from these groups. Comparative genomic analyses are leading to discovery of molecular characteristics that are specific for different groups of Bacteria and Archaea. These markers include conserved inserts and deletions in universal proteins and lineagespecific proteins, which provide novel means for identifying and circumscribing these groups of prokaryotes in clear molecular terms and for understanding their evolution. Because of their taxa specificities, further studies on these newly discovered molecular characteristics should lead to discovery of novel biochemical and physiological characteristics that are unique to different groups of microbes. The focus of my project was phylogenomic studies for two large prokaryotic group: Actinobacteria and Archaea. My goals were to a) identify molecular markers that are specific to Actinobacteria and Archaea at different taxonomic levels, which will help to understand the phylogenetic relationship within these two major groups; b) understand the functional significance of Actinobacteria-specific proteins. By comparative genomics approach, a number of conserved indels in various proteins (viz. Coxl, GluRS, CTPsyn, Gft, GlyRS, TrmD, Gyrase A, SahH and SHMT) have been identified that are specific for all Actinobacteria and additional indels were found to be unique to its major subgroups, such as Corynebacterineae, Bifidobacteriaceae, etc. In parallel, a large number of proteins were discovered to be restricted to Actinobacteria at different phylogenetic depths. These identified conserved indels and proteins for the first time provide useful markers for defining and circumscribing the Actinobacteria phylum or its subgroups in clear molecular terms. Similar comparative genomic studies have been carried out on Archaea and a vast number of proteins have been identified that are unique to Archaea or its various lineages. Lastly, I have performed functional studies on one of the Actinobacteria-specific proteins (ASPl). The structure of ASPl was determined and structural comparison indicates that the function of this protein might be novel since it does not match any known protein with or without known function. </p> / Thesis / Doctor of Philosophy (PhD)

La compréhension de l'évolution des génomes est un des enjeux clé de la biologie actuelle. Nous avons rédigé une revue littéraire consacrée à la contribution de la génomique dans la compréhension de la diversité, de l'évolution et des phénotypes d'un super-phylum bactérien, le super-phylum PVC (Planctomycetes, Verrucomicrobiae et Chlamydiae). Ces bactéries proviennent d'environnements variés et présentent des caractéristiques phénotypiques intéressantes. Les analyses génomiques ont révélé la grande diversité de ces espèces, mais ont aussi permis de reconstruire l'évolution de leurs génomes et d'expliquer l'apparition de certains phénotypes particuliers. Une partie de notre travail était consacré à l'étude de l'évolution et de l'impact de la présence d'un plan cellulaire particulier chez les bactéries PVC. Ce plan cellulaire est sujet a différentes interprétations et induirait la compartimentation des cellules en deux régions distinctes. Les résultats obtenus semblent indiquer que cette caractéristique n'induit pas une protection des génomes bactériens vis à vis des transferts de gènes horizontaux, comme on pourrait le supposer. En revanche les observations microscopiques réalisées sur deux espèces ont permis de mieux appréhender l'évolution de ce plan cellulaire. Nous avons, de plus, détecté une contribution de l'environnement concernant la sélection des gènes transférés. Il semblerait que les gènes transférés soient en effet sélectionnés selon leurs fonctions par les différents environnements.Nos travaux ont donc permis d'améliorer la compréhension des relations entre l'évolution, les phénotypes et l'environnement, en particulier chez les bactéries du super-phylum PVC. / The comprehension of genomes evolution is a key issue of modern biology.We wrote a review dedicated to the genomic contribution in comprehension of diversity, evolution and phenotypes, in a bacterial super-phylum named PVC (for Planctomycetes, Verrucomicrobiae and Chlamydiae). These bacteria are distributed in varied environments and present specific phenotypic characteristics. Genomic analyzes revealed the important diversity of these species and allow also to reconstruct the genomes evolution and, in some cases, to explain the presence of specific phenotypes. One part of our work was dedicated to the study of evolution and impact of one of this phenotype, the special cell plan detected in PVC bacteria. This original cell plan is subject to different interpretations and induces the compartmentalization of cells in two different regions, whom one containing the nucleoid. Our results indicate that this feature has probably no role in the protection of bacterial genomes against horizontal genes transfers, so, its function is still unknown. Microscopic observations of two species from PVC super-phylum permit to better understand the evolution of the special cell plan. The environment seems to contribute in the genomes evolution, by selection of genes transferred. Genes transferred are probably selected according to their functions by the different environments.Our works allowed to improve the knowledge about relations between evolution, genomes, phenotypes and environment, especially in bacteria from PVC super-phylum.

In the summer of 2022, a co-assembly of metagenome was created using the microbes found at Barres Woods in Harvard Forest. 14 samples were taken, and sample was split into the organic and mineral layer, which totals 28 Bulk MAGs. Within this Co-assembly, 4 different genomes were found which were designated with the phylum of FCPU426. Three of which were considered medium quality and one being assigned high quality. The novel phyla first appeared in NCBI and GTDB databases in June 2018. The name FCPU426 dates to 2010 and was named based on the 16s amplicon sequencing. The novel phylum is shown to have common ancestors with the phyla of Goldbacteria and Firestonebacteria. Goldbacteria was determined to have the ability to break down cellulose to further enhance carbon cycling. In this analysis of FCPU426, the taxonomy of the phyla will be further investigated. FCPU426 was then annotated to determine if any of the cellulase genes are shared between itself and Goldbacteria. Further annotations were done to spot any genes of interest that create any distinctions between any of its counterparts.

In den Firmicutes wird der Induktorausschluss (Katabolitrepression) durch das am Serin46 phosphorylierte HPr (PTS) vermittelt. Der genaue Mechanismus war jedoch unklar. Um diese Frage auf der Grundlage von isolierten Proteinen zu klären, wurde ein zum Escherichia coli Maltose-/Maltodextrin-ABC-Transporter homologes System aus Lactobacillus casei BL23 (MalE1-MalF1G1K12) als Modellsystem genutzt. Im Rahmen der Promotion wurde über isothermale Titrationskalorimetrie und Fluoreszenzspektroskopie gezeigt, dass das Bindeprotein MalE1 lineare und zyklische Maltodextrine, aber keine Maltose bindet. Experimentell ermittelte dreidimensionale Strukturen von MalE1 im Komplex mit diesen Zuckern belegten eine vergleichbar geschlossene Konformation und dienten zusätzlich als Grundlage, um die fehlende Maltosebindung zu erklären. Die Stimulierung der ATPaseaktivität des in Liposomen und Nanodiscs eingebauten Komplexes wurde jedoch hauptsächlich durch eine MalE1-Beladung mit linearen Maltodextrinen bewirkt. Eine bis zu 85 %ige Inhibierung der ATPaseaktivität durch P-Ser46-HPr belegte erstmals in vitro eine Interaktion von mehr als einem phosphorylierten Protein mit dem Transporter. Analog zum EIIAGlc-Inhibitor des homologen Systems aus E. coli wurden über Quervernetzungsexperimente und massenspektrometrische Analysen Interaktionen mit dem MalK1-Dimer als interagierende Komplexeinheit in der Nähe des Walker A-Motivs nachgewiesen. Über Fluoreszenzmessungen in Anwesenheit des ATP-Analogons TNP-ATP wurde eine unbeeinflusste ATP-Bindung und damit eine fehlende Blockade der γ-Phosphatbindestelle des Walker-A Motivs durch die Phosphorylgruppe von P-Ser46-HPr bestimmt. Die folgende Substitution verschiedener positiv geladener MalK1-Reste, die als potenzielle Interaktionsstellen für die Phosphorylgruppe fungieren könnten, identifizierte K63 in der Nähe des Walker A-Motivs als ersten möglichen Partner. Der genaue Mechanismus der Inhibierung bleibt jedoch unklar. / Catabolite repression is a global mechanism which controls the utilization of carbohydrates in bacteria. In Firmicutes HPr, a component of the phosphoenolpyruvate carbohydrate phosphotransferase system, prevents the uptake of less preferred sugars but only when it is phosphorylated at serine46. However the exact mechanism was unclear. To address this question the purified ATP-binding cassette transporter from Lactobacillus casei BL23 (MalE1-MalF1G1K12) was used as a model system, which is homologous to the Escherichia coli maltose/maltodextrin ABC importer. Isothermal titration calorimetry and fluorescence spectroscopy revealed that the binding protein MalE1 binds linear and cyclic maltodextrins but not maltose. Experimentally determined three-dimensional structures from MalE1 in complex with these sugars show a comparably closed conformation and served as a basis to explain the lack of maltose binding. The stimulation of the ATPase activity of the transporter incorporated in liposomes and nanodiscs however, was mainly caused by MalE1 loaded with linear maltodextrins. For the first time an inhibition of ATPase activity by P-Ser46-HPr up to 85 % and an interaction of more than one phosphorylated protein with the transporter was demonstrated. Analogous to the EIIAGlc inhibitor of the homologous system from E. coli, cross-linking experiments and mass spectrometric analyzes revealed interactions with the MalK1 dimer near the Walker A motif. Fluorescence measurements in the presence of the ATP analogue TNP-ATP, however, revealed an unaffected ATP binding and thus a lack of blockade of the γ-phosphate binding site (Walker A motif) by the phosphoryl group from P-Ser46-HPr. The following substitution of several positively charged MalK1 residues that could act as potential sites of interaction for the phosphoryl group, identified K63 near the Walker A motif as the first potential partner. The exact mechanism of inhibition, however, remains unclear.

Horizontal gene transfer (HGT) is probably the most important mechanism for functional novelty and adaption in bacteria. However, a robust understanding of the rates of HGT for most bacterial species and the influence of the ecological settings on the rates remain elusive. Four whole-genome comparative studies of free-living bacteria will be described that integrated physiological and ecological data with novel detection bioinformatic pipelines to elucidate the magnitude of HGT at three distinct levels of genetic relatedness: i) the species level, where overlapping ecological niche among co-occurring bacteria in the water column of the Baltic Sea has caused HGT to have been so rampant that it has served as the force of species cohesion; ii) the genus level, where HGT appeared to predominantly mobilize a limited number of genes with ecological/selective advantage (e.g., antibiotic resistance genes) among distinct pathogenic Campylobacter species and hence, did not lead to species convergence; and iii) the phylum level, where HGT was found to be, in general, less frequent than the genus level but, over evolutionary time, has assembled a large part of the metabolic functions of natural microbial communities, especially within organic matter rich, anaerobic habitats. In conclusion, this work advances the methods to link ecological relationships with HGT and suggests that HGT among very divergent organisms may have been more frequent than previously thought and led to successful adaptation.

This diploma thesis deals with the topic of phylum cnidaria as it is being taught in biology classes at primary schools and grammar schools. This work is divided into three parts. The first part deals the position of the topic of phylum cnidaria among curricular documents. It states its classification in the Czech Framework education programme (for Elementary Education) and some chosen School Education Programme. Furthermore, this part analyses the topic as it is written in biology class books, with the clause by the Ministry of Education, Youth and Sports of the Czech Republic. Representatives of phylum cnidaria that are mentioned in biology class books are characterised more thoroughly within the scope of the analysis. The second part of the thesis deals with the teaching methods and forms that are used in teaching biology the most frequently and the questionnaire research that tells us which forms and methods are being used the most by teachers to teach about phylum cnidaria and what specific activities they employ in class. The last part of the thesis consists of suggestions for practical activities for teaching the topic of phylum cnidarian inspired by the results of the survey and a description of educational excursion to the World of Jellyfish aquarium with 6th grade pupils. KEYWORDS phylum...

A abundância e diversidade dos microrganismos do solo podem ser influenciadas por grande número de fatores, associados, principalmente, ao tipo de solo, sua cobertura e uso. A microbiota do solo apresenta grande importância pelos processos desempenhados pelos seus organismos, essenciais para todos os ecossistemas terrestres. Apesar da grande complexidade associada a esses estudos, os fatores que influenciam as comunidades microbianas podem ser melhor elucidados pela pesquisa com grupos específicos. Nesse sentido, o filo bacteriano Verrucomicrobia, grupo ubíquo em solos, apresenta elevada abundância em diferentes ambientes, o que sugere sua grande importância ecológica. Mas, pela dificuldade de isolamento de seus organismos, ainda pouco se sabe a respeito da ecologia do filo. O presente trabalho foi desenvolvido com o objetivo de analisar os efeitos do uso do solo e da rizosfera de cana-de-açúcar nas comunidades de Bacteria e do filo Verrucomicrobia. Com essa finalidade, amostras de solo foram coletadas em áreas sob diferentes usos em uma usina sucroalcooleira na cidade de Piracicaba (SP). As amostras foram utilizadas em dois estudos distintos. No primeiro, foram analisadas as comunidades microbianas presentes nas amostras de solo obtidas na coleta das áreas de estudo e, no segundo, retiradas de um experimento controlado conduzido em estufa para analisar o efeito do uso do solo e, principalmente, da rizosfera nas mesmas. As comunidades foram avaliadas quanto a sua abundância, pela técnica de qPCR, e estrutura, pela técnica de T-RFLP. No primeiro estudo, a diversidade do filo também foi acessada pelo desenvolvimento de bibliotecas do seu gene 16S rRNA. Os resultados dos estudos indicam que a comunidade bacteriana e, principalmente, do filo foram afetadas pelo uso do solo e pelo manejo adotado em cada área, o que evidencia a importância de sistemas conservacionistas. A análise das bibliotecas demonstrou que o filo Verrucomicrobia apresentou maior diversidade na mata nativa do que nos canaviais. Adicionalmente, a incidência e a abundância das subdivisões do grupo foram alteradas de acordo com o uso do solo. As comunidades também foram influenciadas pela rizosfera em sua estrutura e abundância, resultados de grande interesse para o estudo do filo, pois poucos trabalhos analisaram o efeito rizosférico em seus organismos. Além disso, a sua elevada abundância encontrada no estudo, que tem sido comumente subestimada, ressalta a importância de trabalhos com foco específico em grupos de interesse / The abundance and diversity of soil microbial communities can be influenced by many factors, mainly associated with soil type, its coverage and use. The soil microbiota has great importance due to the processes performed by its organisms, essential for all terrestrial ecosystems. Despite the great complexity associated with these studies, the factors that affect the microbial communities can be better explained through the research of specific groups. In this sense, the bacterial phylum Verrucomicrobia, a ubiquitous soil group, presents high abundance in different environments, which suggests its great ecological importance. But due to the difficulty of isolating its organisms, little is known about the ecology of the phylum. The present work was developed with the objective of analyzing the effect of land use changes and sugarcane rhizosphere on the structure and abundance of Bacteria and Verrucomicrobia communities. For this purpose, soil samples were collected in areas under different land uses in a sugarcane mill in Piracicaba (SP). The samples were used in two separate studies. In the first, the microbial communities present in soil samples obtained in the sampling areas were analyzed and, in the second, taken from a controlled experiment conducted in a greenhouse to analyze the effect of land use and, especially, the rhizosphere on the communities. The communities were evaluated for their abundance, by the qPCR technique, and structure, by T-RFLP. In the first study, the phylum diversity was also accessed with the development of 16S rRNA gene libraries. The results from the studies indicate that bacterial and, especially, the phylum community were affected by land use and the management adopted in each area, which evidences the importance of conservationist systems. The analysis of the clone library demonstrated that the phylum Verrucomicrobia presented greater diversity in native vegetation than in the sugarcane fields. Additionally, the incidence and abundance of the group subdivisions have been changed in accordance with land use. The structure and abundance of the communities were also influenced by the rhizosphere, results of great interest to the reserach of the phylum, since few studies have analyzed the rhizosphere effect on its organisms. Furthermore, the high abundance of the phylum found in the study, which has been commonly underestimated, emphasizes the importance of research with specific focus on groups of interest

A abundância e diversidade dos microrganismos do solo podem ser influenciadas por grande número de fatores, associados, principalmente, ao tipo de solo, sua cobertura e uso. A microbiota do solo apresenta grande importância pelos processos desempenhados pelos seus organismos, essenciais para todos os ecossistemas terrestres. Apesar da grande complexidade associada a esses estudos, os fatores que influenciam as comunidades microbianas podem ser melhor elucidados pela pesquisa com grupos específicos. Nesse sentido, o filo bacteriano Verrucomicrobia, grupo ubíquo em solos, apresenta elevada abundância em diferentes ambientes, o que sugere sua grande importância ecológica. Mas, pela dificuldade de isolamento de seus organismos, ainda pouco se sabe a respeito da ecologia do filo. O presente trabalho foi desenvolvido com o objetivo de analisar os efeitos do uso do solo e da rizosfera de cana-de-açúcar nas comunidades de Bacteria e do filo Verrucomicrobia. Com essa finalidade, amostras de solo foram coletadas em áreas sob diferentes usos em uma usina sucroalcooleira na cidade de Piracicaba (SP). As amostras foram utilizadas em dois estudos distintos. No primeiro, foram analisadas as comunidades microbianas presentes nas amostras de solo obtidas na coleta das áreas de estudo e, no segundo, retiradas de um experimento controlado conduzido em estufa para analisar o efeito do uso do solo e, principalmente, da rizosfera nas mesmas. As comunidades foram avaliadas quanto a sua abundância, pela técnica de qPCR, e estrutura, pela técnica de T-RFLP. No primeiro estudo, a diversidade do filo também foi acessada pelo desenvolvimento de bibliotecas do seu gene 16S rRNA. Os resultados dos estudos indicam que a comunidade bacteriana e, principalmente, do filo foram afetadas pelo uso do solo e pelo manejo adotado em cada área, o que evidencia a importância de sistemas conservacionistas. A análise das bibliotecas demonstrou que o filo Verrucomicrobia apresentou maior diversidade na mata nativa do que nos canaviais. Adicionalmente, a incidência e a abundância das subdivisões do grupo foram alteradas de acordo com o uso do solo. As comunidades também foram influenciadas pela rizosfera em sua estrutura e abundância, resultados de grande interesse para o estudo do filo, pois poucos trabalhos analisaram o efeito rizosférico em seus organismos. Além disso, a sua elevada abundância encontrada no estudo, que tem sido comumente subestimada, ressalta a importância de trabalhos com foco específico em grupos de interesse / The abundance and diversity of soil microbial communities can be influenced by many factors, mainly associated with soil type, its coverage and use. The soil microbiota has great importance due to the processes performed by its organisms, essential for all terrestrial ecosystems. Despite the great complexity associated with these studies, the factors that affect the microbial communities can be better explained through the research of specific groups. In this sense, the bacterial phylum Verrucomicrobia, a ubiquitous soil group, presents high abundance in different environments, which suggests its great ecological importance. But due to the difficulty of isolating its organisms, little is known about the ecology of the phylum. The present work was developed with the objective of analyzing the effect of land use changes and sugarcane rhizosphere on the structure and abundance of Bacteria and Verrucomicrobia communities. For this purpose, soil samples were collected in areas under different land uses in a sugarcane mill in Piracicaba (SP). The samples were used in two separate studies. In the first, the microbial communities present in soil samples obtained in the sampling areas were analyzed and, in the second, taken from a controlled experiment conducted in a greenhouse to analyze the effect of land use and, especially, the rhizosphere on the communities. The communities were evaluated for their abundance, by the qPCR technique, and structure, by T-RFLP. In the first study, the phylum diversity was also accessed with the development of 16S rRNA gene libraries. The results from the studies indicate that bacterial and, especially, the phylum community were affected by land use and the management adopted in each area, which evidences the importance of conservationist systems. The analysis of the clone library demonstrated that the phylum Verrucomicrobia presented greater diversity in native vegetation than in the sugarcane fields. Additionally, the incidence and abundance of the group subdivisions have been changed in accordance with land use. The structure and abundance of the communities were also influenced by the rhizosphere, results of great interest to the reserach of the phylum, since few studies have analyzed the rhizosphere effect on its organisms. Furthermore, the high abundance of the phylum found in the study, which has been commonly underestimated, emphasizes the importance of research with specific focus on groups of interest