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Flow cytometry in insects biosystematics
Javůrková, Jaroslava ; Šípek, Petr (advisor) ; Sadílek, David (referee)
The size of the genome, in other words the content of DNA in the gametic cells of organisms, is expressed using the C-value indicating the number of nuclear base pairs in units of Mpb or the mass of DNA in units of pg. A simple method used, among other things, to determine the size of the genome is flow cytometry, which in recent years has been very popular, especially among botanists. The study of insects is lagging behind in regard to the number of described species of insect numbering more than one million in comparison to the number of species whose genome size is known (only a few hundred of them). This diploma thesis therefore aims to expand the dataset on the size of the genomes of representatives of the Polyneoptera group, to compare the data found within individual orders and to help understand the links of genome size to the evolution and ecology of insects. This work provides genome size information for eighty species of insects of the Polyneoptera group. The results showed the largest insect genome measured so far - the locust Dictyophorus griseus (149.33 pg), whose genome size exceeds the previously published results by several times. Key words: flow cytometry, genome size, c-value, Polyneoptera
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Genome size variability and its relationship towards selected biological characteristics of arachnids
Šilhavá, Veronika ; Opatová, Věra (advisor) ; Urfus, Tomáš (referee)
Genome size, also called C-value, describes the amount of DNA contained in haploid set of chromosomes in organism. Knowledge of this value can be useful for example as indicator of technical difficulty and financial cost of projects using genome seqencing, or it can be used for taxonomic purposes as well. Genome size encompasses immense variability across the animal kingdom. We also encounter variability among arachnids, even between species within the same genus. However, the relationship between genome size and biological characteristics of arachnids has not been examined so far. In this thesis, I assembled genome size data for all arachnid orders alongside with a variety of their biological characteristics. These traits were subsequently mapped onto the phylogenetics trees. In certain arachnid orders, the genome size could be related to the body size, life span and potentially also to ecology (e.g. web type, retreat). Unfortunatelly, genome size data are available only for a small portion of Arachnida diversity, which complicates the interpretation.
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Genome size variation in microalgae and its evolutionary consequences
Čertnerová, Dora
Eukaryotic organisms exhibit tremendous variability in genome size with no apparent connection to their biological complexity. Although this variation is known to correlate with numerous phenotypic traits, its evolutionary consequences remain widely unknown. This particularly applies to microalgae, where the genome size estimation is often methodologically challenging. Yet, microalgae represent a promising model group to study genome size evolution owing to their lower body complexity, short generation time and large population sizes, the latter two allowing them to quickly respond to environmental challenges. The main aim of this thesis was to enhance our understanding of genome size variation in microalgae and its evolutionary consequences. To do so, together with my co-authors, I summarized the flow cytometry (FCM) protocols used for microalgae and microorganisms possessing small genomes and addressed their limitations resulting mainly from insufficient amounts of biomass, difficulties with nuclei extraction and prominent background noise due to presence of various pigments and secondary metabolites. Further, I provided best practice recommendations that include, among others, analysing young cultures, avoiding long-term cultivation, and testing different isolation buffers and nuclei isolation...
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Genome size variation in microalgae and its evolutionary consequences
Čertnerová, Dora ; Škaloud, Pavel (advisor) ; Boenigk, Jens (referee) ; Bureš, Petr (referee)
Eukaryotic organisms exhibit tremendous variability in genome size with no apparent connection to their biological complexity. Although this variation is known to correlate with numerous phenotypic traits, its evolutionary consequences remain widely unknown. This particularly applies to microalgae, where the genome size estimation is often methodologically challenging. Yet, microalgae represent a promising model group to study genome size evolution owing to their lower body complexity, short generation time and large population sizes, the latter two allowing them to quickly respond to environmental challenges. The main aim of this thesis was to enhance our understanding of genome size variation in microalgae and its evolutionary consequences. To do so, together with my co-authors, I summarized the flow cytometry (FCM) protocols used for microalgae and microorganisms possessing small genomes and addressed their limitations resulting mainly from insufficient amounts of biomass, difficulties with nuclei extraction and prominent background noise due to presence of various pigments and secondary metabolites. Further, I provided best practice recommendations that include, among others, analysing young cultures, avoiding long-term cultivation, and testing different isolation buffers and nuclei isolation...
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Comparative methods for studying adaptive traits of fungal symbionts
Veselská, Tereza ; Kolařík, Miroslav (advisor) ; Voigt, Kerstin (referee) ; Dvořák, Miloň (referee)
The kingdom Fungi encompasses an estimated 2.2 to 6.2 million species that occupy diverse environments, including aquatic, extremely dry, and hot or frosty habitats all over the world. To cope with adverse environmental conditions, fungi have developed numerous adaptations and life strategies, including symbiosis with other organisms, ranging from close, reciprocally beneficial (mutualistic) associations to severe pathogenic infestations. These interactions have an enormous impact on ecosystem functioning, with implications for agriculture and human health. For this reason, understanding the mechanisms enabling the successful development of fungal interactions is necessary for their efficient management. Recent advances in different 'omics' approaches have enabled us to compare species responses to the environment in a more complex way than before and to gain deeper insights into the adaptive mechanisms underlying specific life strategies. My thesis is divided into four main sections. In the first section, I sum up findings about adaptations of fungal symbionts of plants and animals. Then, I introduce two fungal genera, Geosmithia and Pseudogymnoascus, to which I applied comparative methods for tracking adaptive traits. The ecological diversity of the genus Geosmithia allows to trace adaptive...
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An assessment of interspecific hybridization between Central-European taxa from the genus Nymphaea: insights from flow cytometry and multivariate morphometrics
Kabátová, Klára ; Suda, Jan (advisor) ; Rybka, Vlastimil (referee)
Although the genus Nymphaea (waterlily) includes only two native species (N. alba and N. candida) in Central Europe, it poses a great challenge to taxonomy and biosystematics. The determination of both species is hampered by their phenotypic similarities, and species boundaries can be further blurred by interspecific hybridization. In addition, ornamental cultivars of different parentage often escape from cultivation and make the situation even more complex. To get insight into the caryological and phenotypic variability of czech waterlilies, the DNA flow cytometry and both distance-based and geometric morphometrics were used. Collections showed two different groups of fluorescence intensities, corresponding to N. alba and N. candida, respectively. In addition, intermediate values of nuclear DNA amount were found in some plants from South Bohemia, indicating their hybrid origin. Surprisingly, ornamental cultivars possessed the smallest genome sizes. The amount of nuclear DNA therefore seems to be a promising species-specific marker that enables not only native species but also cultivars to be distinguished. Cytometrically-proven individuals have been subjected to multivariate morphometrics, and high differenciation was discovered especially between native species. More complicated seems the distinction...
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Flow cytometry and its use for study of insects
Stuchlíková, Magdalena ; Janšta, Petr (advisor) ; Sadílek, David (referee)
Flow cytometry is a modern technique in research, playing a significant role in biomedicine and botanics. Despite its benefits (speed, simplicity, low costs), flow cytometry is currently not used in the study of insects on a large scale. This thesis gives an overview as to how flow cytometry is used in research on insects and summarises the results of such study. This pertains to genome size and its connections to other phenomena. Other focal points of research, such as ploidy and base pair ratios, are addressed to a lesser extent. Key words: flow cytometry, use in study of insects, genome size, ploidy, invertebrates
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