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A simple method for detection of Czech bolting garlics (Allium sativum L.) using microsatellite analysis: methodology for practice
Ovesná, Jaroslava ; Mitrová, Katarína ; Kučera, Ladislav
The aim of the methodology was to verify and compile a set of microsatellite markers for the purpose of verification to be used for the undisputed identification of selected types of Czech bolting garlics. The object of submitted methodology is to simplify procedure that allow to determine Czech bolting garlic using microsatellite analysis (SSR- Single Sequence Repeats), which is applicable in testing laboratories. The described method allows to identify the variety and breeding material of bolting garlic of Czech origin using DNA markers (microsatellite alleles) that are present only for this group of varieties and breeding lines. By the amplification of the genome containing the microsatellite locus by polymerase chain reaction (PCR) using specific, fluorescently labeled primers, and subsequent analysis of the lengths of PCR products. Methodology newly brings a unique set of diagnostic SSR markers, describe the evaluation and interpretation of the results.
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Genetic diversity and evolutionary history of Czech endemic taxa from the genus Dianthus
Vítová, Jana
This MSc. thesis deals with karyological, genetic and phenotypic variation of selected taxa from the genus Dianthus L. in the Czech Republic and adjacent countries. The evolutionary history of the genus has been shaped by several microevolutionary processes, including interspecific hybridization, genome duplication, and edaphic speciation. These processes led to the origin of a number of phenotypically similar taxa, which are often restricted to a narrow geographic area. One subendemic and three endemic taxa occur in the Czech Republic. These (sub)species were used as model groups to gain insight into microevolutionary processes in small populations and the postglacial development of the genus in Central Europe. The thesis consists of three parts, each addressing different evolutionary phenomenon: Dianthus arenarius subsp. bohemicus is a critically endangered endemic psammophyte currently known from a single population in Central Bohemia. The site is also inhabited by widespread D. carthusianorum. Interspecific hybridization has been suspected on the basis of morphological characters, but this has never been confirmed by any other technique. I exploited differences in the number of chromosomes between both species and, with the aid of DAPI flow cytometry, estimated relative DNA contents of most...
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Phenotypic variability and evolutionary relationships among populations of Primula elatior along an altitudinal gradient
Konečná, Veronika ; Kolář, Filip (advisor) ; Mandák, Bohumil (referee)
Primula elatior represents a promising model for addressing the role of phenotypic plasticity vs. genetic basis of traits in populations differentiated along altitudinal gradients. Phenotypic plasticity is mainly exhibited by the extensive variability of the leaves. Genetic structure of P. elatior populations has been poorly studied so far, the first insights suggested potentially large intraspecific genetic variation. Therefore, the first aim of this study was to reveal the genetic structure as well as morphological and cytological variation of P. elatior populations in the central Europe and compare those to another two intraspecific taxa from the southeastern Europe. Further, I selected three target mountain regions to address the role of altitudinal differentiation for structuring genetic diversity. Finally, I evaluated the status of the putative endemic species Primula elatior subsp. corcontica from the Krkonoše Mts. Microsatellite analyses, multivariate morphometrics and flow cytometry were employed to evaluate variation of P. elatior populations. I identified six major genetic groups by genotyping 12 nuclear microsatellite loci in the study area of central and southeastern Europe with the clear separation of subp. intricata. Focused on the central Europe, three well-defined genetic groups...
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Hybridization of crab apple with domestic apple trees
Portl, Jiří ; Fér, Tomáš (advisor) ; Prančl, Jan (referee)
This bachelor thesis summarizes known information about hybridization (anthropohybridization) crab apple with orchard apple. Next it follows up mainly genetic and morphometric methods that haven't been used in the hybridization studies yet. Crab apple (Malus sylvestris) is the only one wild species in Malus genus in Europe. However in the last few decades these populations are endangered by orchard apple (Malus ×domestica), that originates from central Asia. It often outspreads into the free nature and thereby closer to wild apple plants, that leads to easier hybridization of these two woods. This process leads up to decreasing number of genetically pure crab apple individuals and also to slow hybrid plants expansion. This trend, which was called anthropohybridization, can be observed also in the Prunus genus, where there hybridizes wild species European dwarf cherry Prunus fruticosa with tart cherry Prunus cerasus. Recent studies though refers to following genetic aspects of Malus domestica, namely formation by triple hybridisation. Several locations of crab apple were observed and analysed in Europe in the last few years for study of anthropohybridization degree with Malus ×domestica. There were observed hybrid plants in common with genetically pure Malus sylvestris. Key words: crab apple,...
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Ecological consequences of polyploidization in populations of the wetland plant Butomus umbellatus
Rydlo, Jan ; Fér, Tomáš (advisor) ; Trávníček, Pavel (referee)
This thesis aimed on comparison of environmental requirements and morphological and biological traits of diploid and triploid cytotypes of wetland plant flowering rush (Butomus umbellatus), occurring in Eastern Slovak Lowland in unique mixed populations, and to develop primers for variable microsatellite loci suitable for further genetic studies. In total, 72 populations of Butomus umbellatus were found. The analysis of relative genome size by the flow cytometry (FCM) technique revealed 34 exclusively triploid cytotype populations, 8 exclusively diploid populations and 30 mixed populations of Butomus umbellatus. In mixed populations, the numbers of diploid and triploid individuals are quite equivalent and the plants create combined compact communities. Analyses of environmental characteristics of sites with specific cytotypes (PCA, RDA, ANOVA) revealed no difference in site requirements of diploid and triploid Butomus umbellatus plants. Morphometric analyses (PCA, CDA) of traits of 36 diploid and 54 triploid plants confirmed differing morphology of both cytotypes occurring in the nature. Subsequent replanting of these plants into pots and cultivation further increased these differences. The ratio of width and length of internal perianthium is the most reliable parameter to distinguish diploid and...
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Spreading of praying mantis (Mantis religiosa) in Europe
Vitáček, Jakub ; Janšta, Petr (advisor) ; Gvoždík, Václav (referee)
Climate change is one of the most important factor determining species ranges. In Europe there is now evidence for northward areal expansion in many Mediterranean insects including the praying mantis (Mantis religiosa). This species is the only representative of the order Mantodea inhabiting central Europe. The northern edge of the species distribution currently reaches latitude 53ř North. Although, the praying mantis is well known insect there is not enough evidence about its phylogeography. In this work three mitochondrial genes (COI, COII, Cyt b) were selected for phylogenetic study. Results indicate three statistically supported distinct lineages in Europe: Eastern European, Central European and Western European. Presumably these lineages are consistent with isolation during the last glacial and re-colonization from glacial refugia. Reduced haplotype diversity on the northern edge suggests currently established populations at the northern distribution border. To validate mtDNA results it was also considered four microsatellite loci. Due to different type of inheritance mtDNA and nuclear DNA it is possible to compare two independent genetic datasets. Microsatellite analysis confirmed results obtained on mitochondrial data. Three major genetic clusters were found: east, west and central. Spatial...
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Molecular typization of isolates from Arthroderma benhamiae complex, a zoonotic agent of epidemic dermatophytosis in Europe
Čmoková, Adéla ; Hubka, Vít (advisor) ; Hrabák, Jaroslav (referee)
Arthroderma benhamiae has been almost unknown among clinical mycologists but it is a cause of around 40 % of dermatophytic infections according to current studies in the Czech Republic. The species is primarily transmitted to humans from guinea pigs and other rodents. The epidemiological situation is similar in other countries in Central and Western Europe. The reason of significant increase in the incidence of infections has not been identified yet, and no sufficiently informative molecular markers have been developed for typification of the species that could help to resolve the cause of this problem. For the purposes of this study, isolates (n=268) from cases of human and animal dermatophytoses from the Czech Republic, Germany, Belgium, Switzerland, Italy, Japan and USA were selected. Ten variable microsatellite markers were developed and sequence analysis of two genetic loci (ITS rDNA and gpdh gene) were performed to reveal intraspecific variation. Phenotype was also studied at the level of micro- and macromorphology of the strains and growth parameters at several temperatures and on several cultivation media. Mating type idiomorph of each isolate was determined and mating experiments were performed by crossing pairs of genetically related as well as genetically distant strains. Sequence...
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Systematics and phylogeography of diploid lineages of Arabidopsis arenosa complex
Hyklová, Lucie ; Záveská, Eliška (advisor) ; Krak, Karol (referee)
The Arabidopsis arenosa species complex is closely related to the model species Arabidopsis thaliana. Distribution range of the diploid lineage covers most parts of Central and South-eastern Europe with a remarkable diversity of ecological niches from sea level to alpine peaks. There is still little known about genetic relationships among the diploid populations of A. arenosa. The evolutionary history of the diploid cytotype across its entire range was explored by using 14 nuclear microsatellite loci. Five lineages occupying biogeographically distinct regions were identified in Arabidopsis arenosa complex. These lineages were labelled as Western Carpathians, Eastern Carpathians, Pannonian Lowland, Dinaric Alps and Baltic Shore (according to geographical regions where these lineages occur). Powered by TCPDF (www.tcpdf.org)
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Internal structure of the Balkan refugium using Erinaceus roumanicus as a model organism
Eliášová, Kristýna ; Černá Bolfíková, Barbora (advisor) ; Zemanová, Barbora (referee)
The aim of this work was to describe the spatial distribution of genetic variability of the northern white-breasted hedgehog (E. roumanicus), mainly within the Balkan Peninsula and Central Europe, as these areas are crucial for understanding the effects of Pleistocene climate oscillations on the genetic architecture of the species. Based on this spatial distribution, hypotheses about the possible structuring of the Balkan refugia were formulated. A total of 260 individuals and 9 microsatellite loci were used for the analysis. Using the approach of landscape and population genetics several possible isolated subpopulations within the area surveyed were identified. Unique status was proved for a population originating from Crete, probably influenced by mechanisms of island evolution. Differences from the rest of the range were ascertained also in population from the Czech Republic, located in a secondary contact zone with the E. europeaus. The role of interspecific interactions and possible introgression should be considered in this case. In the area of Romania and the area south of the Balkan Peninsula the highest genetic distances between individuals were identified, probably associated with the occurrence of geographical barriers and the possible presence of glacial subrefugia. Other identified...
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On the Origin of Chenopodium album by Means of Hybridization
Kondrysová, Eva ; Mandák, Bohumil (advisor) ; Mráz, Patrik (referee)
Diploma thesis deals with genetic variability and evolutionary relationships among selected species of the group Chenopodium album and is especially focused on the origin of hexaploid species C. album in the Czech Republic. The main aim of the study was detection of putative recent emergence of hexaploid C. album s. str. in mixed populations of diploid (C. ficifolium and C. suecicum) and tetraploid (C. strictum and C. striatiforme) species. To assess phylogenetical relationships I performed an analysis of nuclear microsatellite loci, that are suitable for detection parental lines within polyploid species. As long as microsatellite primers have been developed for studied group, therefore they had to be cross-amplified from closely related species C. quinoa. Three PCR multiplexes were assembled by cross-amplification of microsatellite primers. These PCR multiplexes were tested on representative sample sets to estimate genetic variability of individual microsatellite loci. A total of 911 individuals were analyzed from five localities from the Czech Republic. The results of analyses revealed (1) high interspecific differentiation, (2) gene flow among species of the same ploidy levels, (3) presence of hybrids among diploid species of C. ficifolium and C. suecium as well as among tetraploid species C. strictum...
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