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Evolution of mastigont in Excavata
Táborský, Petr ; Čepička, Ivan (advisor) ; Hampl, Vladimír (referee)
In recent years we have been discovering a huge diversity of eukaryotic organisms. One of the big super-groups is Excavata. Examination of the organisms from this group is fairly important as far as theoretical and practical reasons are concerned. A significant number of authors believe that there is the root of all eukaryotic organisms in this group. And what is more, a lot of studies prove the point. Within Excavata group we can find the organisms with the most primitive mitochondrial genome which is known. Several other representatives are an important human pathogenes. In the first part of the thesis I focus on the historical review of the creation of the super-group Excavata and I also include a part regarding the theory about Archezoa. The next section deals with the general description of mastigont (basal bodies and cytoskeleton that is associated with them). The last part presents mastigont descriptions of some derived representatives of Excavata. Some of these organisms may be free-living and some live inside other organisms.
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Molecular phylogenetics and geometric morphometrics of aerophytic green algae of Coccomyxa/Pseudococcomyxa s.l. complex
Nemjová, Katarína ; Neustupa, Jiří (advisor) ; Hampl, Vladimír (referee)
This thesis deals with molecular diversity and shape variation of the traditionally delimited unicellular coccoid genera Coccomyxa SCHMIDLE and Pseudococcomyxa KORŠIKOV (Trebouxiophyceae, Viridiplantae). The complex of these traditional genera was found monophyletic position within the class Trebouxiophyceae on the basis of 18S rDNA sequences. Subdivision of this lineage into individual clades was based on 18S rDNA, ITS1 and ITS2 sequences. The results did not confirm delimitation of traditional genera. However, they suggest ecological differentiation of individual clades. The results of phylogenetic analyses were further supported by reconstructions of ITS2 secondary structures and analyses of the compensatory base changes (CBC) among individual clades. The mutual relationships among clades remained, however, partially unclear. Geometric morphometric variation of cell shapes illustrated pattern that was mostly not correlated with molecular data. This indicates high degree of evolutionary plasticity and possible unreliability of these morphological features in taxonomy of the group. Powered by TCPDF (www.tcpdf.org)
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Type II secretion system in the mitochondria of Naegleria gruberi.
Krupičková, Alžběta ; Doležal, Pavel (advisor) ; Hampl, Vladimír (referee)
Several types of the transport (secretion) systems can be found in the membranes of gram-negative bacteria. These systems export proteins into the extracellular milieu of bacteria. One of them, type II secretion system (T2SS), exports the folded proteins through the outer bacterial membrane. The core subunits of T2SS were discovered in the genomes of several protists belonging to the Discoba and Malawimonada groups, including Naegleria gruberi and Malawimonas sp. str. 249. Our experiments suggest that these subunits are localized in the mitochondria of these protists. The discovery of these genes in eukaryotes is surprising. If the T2SS was active in these mitochondria, it would represent an evolutionary intermediate stage of the organelle, which can export and import the proteins. In experimental part of the thesis, we studied how the proteins of N. gruberi and M. sp. str. 249 interact. We demonstrated several interactions, which correspond to the function of bacterial T2SS. To this aim, we employed bacterial and yeast two-hybrid systems. Further, we explored the import of the core subunit of T2SS, pseudopilin, of N. gruberi into the inner mitochondrial membrane. And we also investigated the import of M. sp. str. 249 secretin into the outer mitochondrial membrane.
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Pathogenic trichomonads in cats and dogs
Vobořilová, Pavlína ; Tachezy, Jan (advisor) ; Hampl, Vladimír (referee)
Trichomonads are anaerobic flagellated protists that are either parasites or commensals. They frequently inhabit digestive, respiratory, and urogenital tracts of vertebrates, including domestic cats and dogs. In these hosts, four trichomonad species has been described: Tetratrichomonas canistomae and Tetratrichomonas felistomae that are commensals of the host oral cavity; Pentatrichomonas hominis, a commensal of intestinal tract that could be found in dogs and cats but also in other mammals including humans; and pathogenic Tritrichomonas foetus that causes, in addition to cattle infection, feline intestinal trichomonosis. Although, trichomonads in dogs and cats are probably of cosmopolitan distribution we have no information about their presence in Czech Republic. The first aim of this study was to distinguish types of trichomonads present in the oral cavity of dogs and cats and to get preliminary epidemiological data. The second aim was to demonstrate the presence of Tritrichomonas foetus in cats and dogs in the Czech Republic and to identify potential risk factors for the disease. Cultivation and nested PCR were used to determine the presence of trichomonads in dogs and cats. Sequencing and phylogenetic analysis based on ITS1-5.8rRNA-ITS2 gene sequence was used to identify species of isolated...
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Prokaryotic symbionts of free-living anaerobic protists
Poláková, Kateřina ; Čepička, Ivan (advisor) ; Hampl, Vladimír (referee)
Anaerobní prvoci jsou organismy obývající ředí bez přítomnosti kyslíku. Najdeme je anoxických habitatech jako jsou mořské a sladkovodní sedimenty, komunální skládky, nesmíme opomenout zástupce žijící v bachoru přežvýkavců, trávicím traktu švábů a dalších živočichů. Většinou mají anaerobní deriváty mitochondrií, často hydrogenosomy, organely produkující vodík. Metabolismus anaerobních prvoků je ve srovnání s aerobními organismy méně efektivní Časté interakce anaerobních ický my však mohou Symbiózy mezi anaerobními běžné a vznikly u zástupců mnoha anaerobních linií. žít buď endosymbioticky, uvnitř buňky hostitele, nebo ektosymbioticky, na povrchu hostitelské buňky. Dvě ekologicky významné skupiny prokaryot si dokázaly osvojit život symbiotickém vztahu volně žijícími anaerobními prvoky anogenní archebakterie, osídlující hlavně , využívají vodík nické sloučeniny k anu a síran redukující bakterie, žijící hlavně na povrchu buněk, využívají vodík a různé organické sloučeniny k redukci síranu na sulfan. Velmi málo se ví o bližším charakteru těchto vztahů druhové identitě a hostitelské specifitě prokaryotických symbiontů. Další výzkum je nutný pro pochopení fenoménu symbióz v anoxickém světě. Klíčová slova: anaerobní prvoci symbióza anogenní archebakterie síran redukující bakterie
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FISH method and its use in protistology
Frenclová, Martina ; Hampl, Vladimír (advisor) ; Zubáčová, Zuzana (referee)
Fluorescence in situ hybridization (FISH) is a technique that allows the localization and identification of specific sequences of nucleotides in DNA or RNA, which is subsequently visible under the microscope. FISH involves first denaturing the nucleic acids, either using high temperatures or by treatment with denaturing agents such as formamide.After subsequent induction of reasociation, the examined DNA or RNA pairs according to the complementarity rules with the short molecule called the probe, this process is called hybridization. Hybridization occurs in situ, that is within the examined specimen. Probes can be labeled either directly using fluorophores, or indirectly with a hapten, which is a substance having antigenic properties, which is subsequently detected using labeled antibodies or streptavidine. FISH has a large number of applications in molecular biology and medical science. In laboratory research in protistolgii FISH can be used for example to map the chromosomal genes to study the evolution of genome, analysis of nuclear organization or to confirmation of the origin of DNA sequence. Key words: FISH, fluorescence, probe, cell identification, environmental studies
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Evolution of nuclear and plastid genomes in euglenids
Hrdá, Štěpánka ; Hampl, Vladimír (advisor) ; Krajčovič, Juraj (referee) ; Škaloud, Pavel (referee)
Algae form a diverse group of simple photosynthetic eukaryotes of polyphyletic origin. Algae with a primary plastid (Archaeplastida) acquired it by ingesting cyanobacterium, a prokaryote; algae with a complex plastid acquired their plastid by ingesting another eukaryote with a primary or already complex plastid. Algae with a complex plastid are chimeras containing genes derived from the host genome, as well as genes derived from the genome of the endosymbiont, and also genetic material derived from genomes of their previous stable or transient endosymbionts. One of the groups with plastid derived from green algae are euglenophytes. This thesis deals with the genomes of three organisms that represent individual actors in the endosymbiotic process in euglenophytes. These are a heterotrophic host from the class Euglenida, a phototrophic endosymbiont from the class of green algae Prasinophyceae and the resulting phototrophic euglenid from the group Euglenophyceae. Knowledge of their genomes should illuminate the course of endosymbiotic gene transfer (EGT) in the formation of algae with a complex plastid. We annotated the plastid genome of a phototrophic euglenid Eutreptiella gymnastica and published it as the third plastome of Euglenophytes after the iconic and economically important Euglena gracilis...
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Iron-Sulfur cluster assembly in Monocercomonoides exilis
Vacek, Vojtěch ; Hampl, Vladimír (advisor) ; Balk, Janneke (referee) ; Tsaousis, Anastasios (referee)
In the search for the mitochondrion of oxymonads, DNA of Monocercomonoides exilis - an oxymonad isolated from the gut of Chinchilla, was isolated and its genome was sequenced. Sequencing resulted in a fairly complete genome which was extensively searched or genes for mitochondrion related proteins, but no reliable candidate for such gene was identified. Even genes for the ISC pathway, which is responsible for Fe-S cluster assembly and considered to be the only essential function of reduced mitochondrion-like organelles (MROs), were absent. Instead, we were able to detect the presence of a SUF pathway which functionally replaced the ISC pathway. Closer examination of the SUF pathway based on heterologous localisation revealed that this pathway localised in the cytosol. In silico analysis showed that SUF genes are highly conserved at the level of secondary and tertiary structure and most catalytic residues and motifs are present in their sequences. The functionality of these proteins was further indirectly confirmed by complementation experiments in Escherichia coli where SUF proteins of M. exilis were able to restore at least partially Fe-S cluster assembly of strains deficient in the SUF and ISC pathways. We also proved by bacterial adenylate cyclase two-hybrid system that SufB and SufC can form...
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Phylogenetic Position of Genus Polymastix and Its Prokaryotic Symbionts
Kubánková, Aneta ; Hampl, Vladimír (advisor) ; Kolísko, Martin (referee)
Polymastigidae is one of the five families of oxymonads (Metamonada, Preaxostyla). It includes small flagellates inhabiting the digestive tract of insects. Recently obtained molecular phylogenetic data have questioned the monophyly of this group. The type genus of the family Polymastigidae is Polymastix. The phylogenetic position of this genus has not been studied yet, although this information could significantly help to clarify the relationships within this group. This work provides the first sequence data of this genus, which we have obtained using a single- cell genome amplification and subsequent whole genome sequencing on Illumina HiSeq X Ten (Macrogene) from three individuals of P. melolonthae inhabiting the posterior hindgut of crane fly larvae. Phylogenetic analysis based on the gene for 18S rRNA, EF-1α and HSP90 yielded several roughly equally probable hypotheses about the position of this genus within Oxymonadida. The first of them places the genus Polymastix in a close proximity to the genus Streblomastix, according to the second hypothesis, Polymastix is sister to Termitimonas travisae. A characteristic feature of this protist is ectosymbiotic bacteria attached to its surface. These were necessarily amplified and sequenced together with their hosts, which allowed us to obtain sequence...
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