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Evolution of karyotypes and sex determination in the turtle family Geoemydidae
Clemente, Lorenzo ; Rovatsos, Michail (advisor) ; Montiel Jimenez, Eugenia Elisabet (referee) ; Castiglia, Riccardo (referee)
(IN ENGLISH) The majority of studied turtles show temperature-dependent sex determination, but genotypic sex determination (i.e. presence of sex chromosomes) was identified sporadically. This thesis aims to investigate and expand our knowledge on the evolution of the karyotype and the sex determination in turtles, particularly focusing on the family Geoemydidae, a group of turtles with previously documented variability in sex determination systems. The presence of sex chromosomes was explored by a combination of conventional and molecular cytogenetic techniques for the analysis of karyotypes, distribution of constitutive heterochromatin (C-banding) and repetitive elements and comparative genome hybridization (FISH, CGH). In total, 49 species of turtles from nine different families were cytogenetically examined in this study. In the family Geoemydidae, a remarkable similarity in karyotypes was identified, consisting of 2n=52 chromosomes (which is suggested to be the ancestral diploid number for all turtles) and a similar topology of rDNA loci and telomeric repeats. Sharma et al. (1975) previously reported ZZ/ZW sex chromosomes in Pangshura smithii. However, in the analysis presented in this thesis, it is suggested a possible misidentification of these sex chromosomes due to erroneous pairing of...
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RNA directed DNA methylation in Arabidopsis thaliana
Motylová, Šárka ; Fischer, Lukáš (advisor) ; Moravec, Tomáš (referee)
The differential transcriptional activity of the genome is provided by epigenetic modifications, which include DNA methylation, alteration of histone N-terminal amino acids and changes in histone variants. RNA interference is a regulatory process, in which transcriptional or post-transcriptional silencing of exogenous or endogenous sequences is mediated by the action of small RNAs derived from these sequences. The 24-nucleotide siRNAs, forming a fraction of small RNAs, direct de novo DNA methylation and participate in the maintenance of DNA methylation (RNA-directed DNA methylation; RdDM), which facilitates transcriptional silencing of heterochromatin and transposable elements representing a large part of plant genomes. The presence of two RNA polymerases involved in this pathway is characteristic for flowering plants, which were discovered for the first time in the genome of Arabidopsis thaliana, which has also become the main plant model for the study of RdDM. Polymerase IV transcribes siRNA precursors; siRNAs are subsequently associated with AGO4 proteins and guide methylation enzymes to the target sequences via complementarity with polymerase V transcripts.
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Karyotype analysis of selected representatives of two pedipalpid orders, Amblypygi and Uropygi
Sember, Alexandr ; Král, Jiří (advisor) ; Ráb, Petr (referee)
Karyotype analysis of selected species from arachnid orders Amblypygi and Uropygi Whip spiders (Amblypygi) and whip scorpions (Uropygi) represent relict arachnid orders which has been found already at Upper Carboniferous strata. Although cytogenetic data from amblypygids and uropygids might be important to reconstruct karyotype evolution of arachnids, cytogenetics of these orders is almost unknown. Presented study is aimed in analysis of karyotype and meiosis in 16 species of Amblypygi and 4 species of Uropygi. Both groups are characterized by considerable range of diploid chromosome numbers (2n = 24 - 86 in Amblypygi and 36 - 66 in Uropygi). Analysed species does not exhibit morfologically differentiated sex chromosomes. Differentiation of sex chromosomes on molecular level was revealed in amblypygid Paraphrynus mexicanus by comparative genome hybridization. Obtained data indicate XY/XX sex chromosome system in this species. Comparison of karyotype data indicates reduction of chromosome numbers during evolution of both orders. In Amblypygi, this reduction was accompanied by increase of number of biarmed chromosomes. This trend is not apparent in Uropygi. Karyotypes of most analysed amblypygids and uropygids are also characterized by low amount of heterochromatin. Most studied species exhibit two pairs...
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Evolution of sex chromosomes and karyotypes in boas and pythons
Charvát, Tomáš ; Rovatsos, Michail (advisor) ; Majtánová, Zuzana (referee)
- ABSTRACT - Snakes (Serpentes) are a group of squamate reptiles (Squamata) that represents more than one third of the total reptile species diversity. Snake karyotype is generally conserved with the most common chromosome number of 36 (16 macro- and 20 microchromosomes) in diploid state. It is believed that this karyotype was also present in the common ancestor of all snakes. The majority of snake species belong to the group Caenophidia and share homologous ZW sex chromosomes. Snakes from the groups "Scolecophidia" and "Henophidia" have mostly poorly differentiated, homomorphic sex chromosomes, which made them impossible to distinguish from the autosomes in the past. These snakes were for many years assumed to have ZW sex chromosomes as well. However, recent studies demonstrated not only ZW but also two non- homologous XY sex chromosome systems in non-caenophidian snakes and thus the sex determination systems in snakes are much more variable than previously thought. In this thesis, eight species of henophidian snakes (representatives from the genera Eryx, Cylidrophis, Python and Tropidophis) and one caenophidian species (Ophiophagus hannah) were examined using conventional and molecular cytogenetic methods. However, sex chromosomes were not detected in the henophidian species, only in Ophiophagus hannah,...
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Evolution of sex chromosomes in reptiles
Mazzoleni, Sofia ; Rovatsos, Michail (advisor) ; Zrzavá, Magda (referee) ; Liehr, Thomas (referee)
- ABSTRACT - Among vertebrates, reptiles represent the ideal group for the study of sex determination. Reptiles include lineages with environmental sex determination (ESD) as seen in crocodiles and tuatara, lineages with genotypic sex determination (GSD), like e.g. iguanas, chameleons, skinks, lacertid lizards and birds, and few groups which possess variability in sex determination mechanisms, i.e. geckos, dragon lizards and turtles. This thesis is focused on the evolution of sex chromosomes and sex determination in turtles. The majority of turtle species exhibit ESD, which is considered the ancestral sex determination system of this group, while GSD either as male or female heterogamety evolved independently at least five times. We investigated the presence of sex chromosomes in representative species of turtles by cytogenetic analyses. The analyses included the reconstruction of karyotypes, distribution of constitutive heterochromatin (C-banding, methylation analysis) and repetitive elements (fluorescence in situ hybridization) and comparative genome hybridization (CGH), which often characterize the degenerated Y or W and can be helpful in the identification of "cryptic" sex chromosomes. We described XX/XY sex chromosomes in seven previously unstudied Australasian chelids (Pleurodira) from the genera...
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Evolution of sex chromosomes and karyotypes in snakes
Augstenová, Barbora ; Kratochvíl, Lukáš (advisor) ; Majtánová, Zuzana (referee)
Snakes (Serpentes) are a group of squamate reptiles (Squamata) including more than 3600 species. The vast majority of snakes are from the group Caenophidia, which includes approximately 90% of all extant snake species and represents the most studied lineage. Squamate reptiles are variable in sex determination and genome organisation, however, caenophidian snakes are quite stable in these respects. The typical - and probably ancestral - snake karyotype is composed of 36 chromosomes with 16 macro- and 20 microchromosomes. In all snake species, genotypic sex determination (GSD) is expected and it was assumed until recently that all snakes possessed ZZ/ZW sex chromosomes. The main reason for this is that most of the studied snakes were from the group Caenophidia and in the rest of the snake lineages it was believed that their sex chromosomes were homologous. In fact, the sex chromosomes of non-caenophidian snakes have not yet been identified - with the single exception of Acrantophis dumerili. Nevertheless, a recent study showed that there was an independent evolution of XX/XY sex chromosomes in pythons and boas. Sex chromosomes of these snakes are homomorphic and so far they have not been detected by classical cytogenetic methods. In this context, the aim of this study is to explore whether it is...
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Heterochromatin variants of the human karyotype
Michalová, Michaela ; Šípek, Antonín (advisor) ; Forman, Martin (referee)
Chromosomes are cell structures consist of chromatin, out of which one kind is a constitutive heterochromatin, which contains non-coding sequences only and is transcriptionally inactive. Heterochromatin blocks consist of highly repetitive sequences of satellite DNAs, which allows the parts to be variable. The largest areas of heterochromatin can be found at long arms of chromosomes 1, 9, 16 and Y, but heterochromatin areas also affect the variability of the short arms of acrocentric chromosomes 13 - 15, 21 and 22. This bachelor thesis based on a number of scientific essays summarizes findings about heterochromatic variants, their occurrence, frequency, possibilities of examining as well as their clinical significance in today's world. It mainly highlights their possible connection with reproductive failures and contradictory results of individual observations. Contemporary results show the importance of new laboratory methods (molecularly cytogenetic), which can be used in specifying and more detailed sorting of findings, which were previously tagged as harmless variants according to less accurate banding methods. The summarization implies that emphasis should be put on the study of clinical meaning of heterochromatic variations. These can influence the progress of meiosis and thus trigger...
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Cytogenetics of selected groups of ray-finned fishes (Actinopterygii): Evolutionary-ecological questions associated with the dynamics of repetitive sequences and the occurrence of polyploidy
Sember, Alexandr ; Ráb, Petr (advisor) ; Marec, František (referee) ; Král, Jiří (referee)
Ray-finned fishes (Actinopterygii) exhibit the greatest biodiversity among vertebrates. The vast majority of extant actinopterygian fish species belong to clade Teleostei - a lineage whose significant evolutionary success might have resulted from a teleost specific whole- genome duplication (TSGD) that occurred at the onset of this group, subsequent to its divergence from the rest of actinopterygian lineages. Despite the growing body of sequenced fish genomes and analyses of their transcriptomes, the largest contribution to understanding fish genomes comes from analyses of DNA content and from cytogenetics. Genomes of ray-finned fishes and especially those of Teleostei exhibit vast diversity and rapid dynamics of repetitive DNA sequences whose variability is reflected in a wide range of fish genome sizes and in the dynamics behind karyotype differentiation. Therefore, ray-finned fishes offer a unique opportunity to study genome variability as a driving force underlying morphological and ecological diversification, evolution and adaptation. Particularly, the mapping of repetitive DNA sequences by means of fluorescence in situ hybridization (FISH) has proven to be a very useful and informative approach during the last two decades and contributed greatly to our understanding of the fish genome...
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RNA directed DNA methylation in Arabidopsis thaliana
Motylová, Šárka ; Fischer, Lukáš (advisor) ; Moravec, Tomáš (referee)
The differential transcriptional activity of the genome is provided by epigenetic modifications, which include DNA methylation, alteration of histone N-terminal amino acids and changes in histone variants. RNA interference is a regulatory process, in which transcriptional or post-transcriptional silencing of exogenous or endogenous sequences is mediated by the action of small RNAs derived from these sequences. The 24-nucleotide siRNAs, forming a fraction of small RNAs, direct de novo DNA methylation and participate in the maintenance of DNA methylation (RNA-directed DNA methylation; RdDM), which facilitates transcriptional silencing of heterochromatin and transposable elements representing a large part of plant genomes. The presence of two RNA polymerases involved in this pathway is characteristic for flowering plants, which were discovered for the first time in the genome of Arabidopsis thaliana, which has also become the main plant model for the study of RdDM. Polymerase IV transcribes siRNA precursors; siRNAs are subsequently associated with AGO4 proteins and guide methylation enzymes to the target sequences via complementarity with polymerase V transcripts.
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