Annual ryegrass (Lolium multiflorum Lam.) can be divided into two cytotypes: diploid (2n = 2x = 14) or tetraploid (2n = 4x = 28). Transforming annual ryegrass from diploid to tetraploid often results in increased seedling vigor, more robust growth and, thus, supposedly greater yield. There has been conflicting research that suggests no difference in yield between diploid and tetraploid cultivars, but livestock producers’ question whether benefits exist to utilizing tetraploid cultivars for grazing cattle. This study sought to determine if annual ryegrass cytotype affected cattle average daily gain. Although there were no differences in average daily gain between cytotypes, there were differences among cultivars. In 2017, Jumbo (4x) produced the greatest weight gain, while in 2018, Tam-90 (2x) generated the greatest gain. This study suggests that there are no advantages attributed to average daily gain by utilizing tetraploid, as opposed to diploid cultivars.

Polyploidization (whole-genome duplication) is widely considered one of the most important evolutionary forces driving the diversification of flowering plants. Polyploids tend to originate recurrently and many plant species retain individuals of two or more different ploidy levels in certain parts of their distributional range of even within their populations. The main aim of this thesis was to address the understudied aspects of polyploid speciation by employing new, convenient methods and/or studying plant model systems with unique features. Difference in monoploid genome size of Tripleurospermum inodorum (Asteraceae) cytotypes provided a unique opportunity for addressing the rate of spontaneous polyploidization in natural populations by enabling the easy distinction of neopolyploid mutants from long-established polyploids in routine flow-cytometric analyses. Repeated ploidy screening in mixed-ploidy populations of annual T. inodorum have been, to our knowledge, the very first attempt to document temporal changes in cytotype composition in situ. In spite of considerable between- year oscillations in cytotype frequencies, both diploids and tetraploids usually persisted locally for several consecutive years. The common incidence of such ploidy mixtures along with a partial fertility of triploid...

[Abstract] This thesis is aimed at better understanding of cytotype co-existence in mixed- ploidy populations with an emphasis on a microevolutionary processes behind it. Our past knowledge was based on a few thoroughly investigated model taxa like Chamerion angustifolium and Heuchera grossulariifolia, but some generalizations seem to be premature in the light of new findings. A detailed research of other taxa included in the thesis showed that polyploid complexes can vary dramatically in their ability to cope with the co-existence of cytotypes in mixed-ploidy popu- lations. Whereas mixed-ploidy populations are virtually lacking in some species (an example being Vicia cracca, Paper III.), ploidy-heterogeneous populations are very common in others, maintained by free mating interactions and the absence of reproductive isolation among cytotypes (e.g. Pilosella echioides, Paper II.). The strenght and cumulative effect of various breeding barriers (both pre- or post- zygotic) govern the position of a particular multi-ploidy complex between these two extremes and co-determine the type of cytotype co-existence in its mixed- ploidy populations. Despite the fact that the number of studies revealing cytotype co-existence has been increasing rapidly, evolutionary background and consequences of such co-...

Mixed-ploidy populations, consisting of multiple cytotypes, are an optimal system for studying genome doubling consequences in plants. The role of frequency dependent selection, known as minority cytotype exclusion principle, is very important in them, but there are many factors limiting this selection. In my thesis, I evaluate the changes in cytotype frequencies and pattern in permanent plots in natural mixed-ploidy populations of three plant species - Butomus umbellatus, Knautia serpentinicola and Tripleurospermum inodorum and I also experimentally analyze partial aspects of frequency dependent selection towards minority cytotype in artificially established mixed-ploidy arrays of two plant species - Arabidopsis arenosa and Tripleurospermum inodorum. Varied changes in frequencies of minority cytotype ware revealed in permanent plots. Usually there was decrease, but in two plots there was increase in minority cytotype frequency and in one case it became dominant. Observed changes depend on initial frequencies of minority cytotype in populations, biological properties of species and particular environment of permanent plot. Relative indexes describing the frequency and the strength of disturbances and also the amount of soil nutrients was used to compare the environment of permanent plots across...

In the present study, I dealt with morphological, cytological, and ecological research on a rare Central-European species Spergularia echinosperma and its comparison with a similar weedy species S. rubra. Existence of two cytotypes of S. echinosperma significantly differing in their morphology was revealed, as well as distinct morphological differences between the two species were found. Moreover, the analyses revealed one possibly hybridogenous population. In addition, both the species and the cytotypes were also proven to display different germination behavior, which I correlate with their individual ecological adaptations.

Genome duplication plays a significant role in plant evolution. Formation of new polyploids is generally considered to be rare. Nevertheless, under natural conditions mixed-ploidy populations occur in relatively large numbers. Only the observations in the cytotype contact zone can identify all the factors affecting the stability or instability of the population. Number of research focusing on study of cytotype coexistence in natural mixed-ploidy populations is still low. As a model system for the study of mechanisms governing cytotype coexistence was chosen the Tripleurospermum inodorum. Research focused on natural mixed-ploidy populations and also on planted mixed-ploidy populations. Permanent plots were located in south, west, northwest and north Bohemia. Field observations were supplemented by cultivation experiments carried out in the greenhouse. Several phenomena were discovered at the level of whole populations. Cytotype distribution in the plot was random. The spatial structure of natural populations was quickly changing even within a single season. Even between single plot evaluations the cytotype ratios varied. Three percent of all plants were triploid hybrids. Most often detected cytotype in soil seed bank was diploid cytotype. The study of population dynamics shows, that tetraploids are...

The presence of triploid individuals, whether hybrids arising from crosses of diploid and tetraploid plants, or spontaneously generated triploid hybrids diploid populations, is increasingly important in plant populations. However, the possible evolutionary role of the triploid hybrids is closely connected to the frequency of their occurrence, to their relative fitness (compared to parental cytotype) and fertility. As a model plant I choosed Tripleurospermum inodorum, in which a relatively frequent occurrence of triploid hybrids was documented in mixed populations of tetraploids and diploids. This annual plant achieves realy quick its reproductive maturity and is easily cultivated, making it an ideal model for cultivation and pollinating experiments. Compared to parental cytotypes, the triploid hybrids of T. inodorum take approximately the same (mostly mediated) fitness values. In vitro conditions were cultivated to measure of germination rates. Germination rate of triploid seeds was comparable to diploid parental cytotype. In the subsequent comparative cultivation, nearly 150 plant individuals (2x, 3x, 4x, and aneuploid cytotype) in the greenhouse showed that the triploids had intermedial values of their parental cytotypes or close to one of the parents. Thus, the vitality of the triploid plants is...

The strength and direction of interactions between organisms vary spatially across the landscape. Traditionally, the focus has been on how trait variation affects the interactions between species. However, differences in abiotic and biotic environmental factors may also alter the distribution, phenology and behavior of the interacting species. To be able to understand why an interaction varies across the landscape, the effects of trait variation has to be separated from the effects of the environmental context. In this thesis, I try to separate the effects of context and trait differences on plant resistance against herbivory, through experimental and observational studies conducted with two cytotypes of the perennial herb Cardamine pratensis and its main herbivore, Anthocharis cardamines. The results show that differences in plant resistance against oviposition under controlled conditions were mainly mediated by flower size; larger flowers were more attractive to the female butterfly. However, among-populations differences in oviposition under natural conditions were not related to the resistance observed under controlled conditions, or to ploidy type, flowering phenology or plant size. Within populations under natural conditions the oviposition patterns by A. cardamines was affected by the plant traits plant size and flowering phenology. The result of this thesis shows that among-population differences in intensity of plant-herbivore interactions were caused by differences in environmental context rather than by herbivore preferences for any phenotypic plant traits, while host plant selection within population was based on plant traits. This suggests that biotic and biotic context can have important effects on the intensity of plant-herbivore interactions. Although genetic traits influenced the outcome of the interaction within populations, it was the environmental context of the populations that determined largely if the interaction took place or not. / <p>At the time of the doctoral defence the following papers were unpublished and had  a status as follows: Paper 2: Manuscript in review in Plos One; Paper 3: Manuscript in review in <em></em>Ecological Entomology; Paper 4: Manuscript</p>

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No. of bitstreams: 0 Previous issue date: 2017-03-03 / Lippia alba (Verbenaceae), é uma espécie herbácea tropical com grande plasticidade fenotípica e genômica, amplamente utilizada na medicina popular. Recentemente, a espécie foi descrita como um novo complexo autopoliploide contendo cinco números cromossômicos (2x=30, 2x=30+8, 3x=45, 4x=60 e 6x=90), e esforços têm sido feitos a fim de entender sua origem e evolução. No presente trabalho, foram descritos perfis cariotípicos mais detalhados da espécie, por meio de mapeamento cromossômico utilizando sondas espécie-específicas e análises do comportamento meiótico e de viabilidade polínica. A partir do sequenciamento genômico de baixa cobertura (IIlumina MiSeq), foram desenvolvidos novos marcadores citogenéticos (denominados CL66 e CL98) os quais foram utilizados para o mapeamento cromossômico em acessos representando os cinco citótipos do complexo. Para a análise meiótica, seis estágios da divisão (metáfase I; anáfase I + telófase I; metáfase II; anáfase II + telófase II) foram quantificados, e aproximadamente, 100 células foram avaliadas para cada estágio. Os mesmos acessos foram avaliados quanto à viabilidade polínica (1.000 grãos de pólen foram quantificados para cada indivíduo). Os resultados da Hibridização Fluorescente in situ (FISH) revelaram que ambas as repetições satélite estão localizadas na porção terminal dos cromossomos. Em geral, a repetição CL98 mostrou um padrão uniforme nos diferentes acessos. Foram observados dois, três, quatro e seis cromossomos marcados em diploides, triploides, tetraploides e hexaploide, respectivamente, revelando que o número de cromossomos marcados variou proporcionalmente, de acordo com o nível de ploidia do acesso. Por outro lado, a repetição CL66 apresentou-se polimórfica. Variações foram observadas entre os acessos, principalmente, entre os indivíduos diploides. Com relação às análises meióticas, alto percentual de irregularidade foi observado nos citótipos poliploides. Entretanto, alguns acessos 2x também mostraram consideráveis erros durante a microsporogênese. Entre as irregularidades encontradas, destacam-se: pareamento cromossômico anormal; segregação cromossômica desigual; cromossomos perdidos; tríades e políades. Os resultados da viabilidade polínica corroboraram os dados da meiose. A partir do conjunto de dados obtidos foi possível concluir que 1) a metodologia para o desenvolvimento de marcadores cromossômicos específicos para L. alba mostrou-se eficiente; 2) as repetições satélite exibiram diferentes comportamentos (estável e dinâmico) no genoma de L. alba; 3) a ocorrência de microsporogênese irregular em diploides, associada à viabilidade polínica, sugerem que os acessos 2x sejam elementos importantes na formação do complexo poliploide e 4) a ampla variação cariotípica observada na espécie pode ser consequência de múltiplos e independentes eventos de duplicação genômica, aliado a rearranjos cromossômicos. Possivelmente, L. alba encontra-se em processo de estabilização do seu cariótipo tornando a espécie, um importante modelo para estudos de poliploides naturais nos trópicos. / Lippia alba (Verbenaceae) is a tropical aromatic shrub with extensive phenotypical and genomic plasticity widely used in traditional medicine. Recently, the species was described as a new natural autopolyploid complex with five distinct chromosome numbers (2x=30, 2x=30+8, 3x=45, 4x=60 and 6x=90). Strides have been done in order to understand the cytotypes origin and species evolution. In this study, a detailed karyotype of L. alba using Fluorescence in situ Hybridization (FISH) with species-specific probes was described. We also report the meiosis behavior and pollen viability in sixty accessions. Using massive parallel sequencing (IIlumina MiSeq platform) new cytogenetic landmarks (CL66 and CL98) were chosen for probing all cytotypes described for the species. For meiotic analysis, the percentage of abnormalities was quantified, evaluating around 100 cells in six stages (metaphase I; anaphase I + telophase I; metaphase II; anaphase II + telophase II). Around 1,000 pollen per accession were used to estimate pollen viability. FISH results revealed that both satDNA arrays are located preferentially on terminal sites of the chromosomes. In general, the CL98 repeat showed a uniform pattern in different accessions. We observed 2, 3, 4, and 6 marked chromosomes respectively in diploid, triploid, tetraploid and hexaploid accessions revealing that the number of depicted chromosomes varied proportionally according to the ploidy level. On the other hand, the CL66 repeat was polymorphic. Great variations were observed among the accessions mainly within the diploids. In general, the meiotic analysis revealed higher index of abnormalities in polyploid cytotypes. However, some 2x accessions also showed considerable irregularities during the microsporogenesis. Desynapsis, unequal segregation, lost chromosomes, triads and polyads were the most common irregularities observed. Pollen viability analysis corroborated the meiosis data. It was possible to conclude that 1) the development of specific landmarks for L. alba was efficient; 2) the karyotypic profiles of both satDNA revealed different behavior; 3) microsporogenesis analysis and pollen viability of 2x accessions suggest that diploids are the key point for the origin of the polyploid complex and 4) independent and multiples events of genome duplication associated to chromosome rearrangements may have generated great karyotypic variation in the species. L. alba karyotype is possibly under stabilization process making the species an important model to study natural polyploids in the tropics.

Polyploidization belong to principal evolutionary mechanisms in plants. Although it is rather rare in the family Apiaceae, there are several interesting polyploid complexes here. The present study deals with Libanotis pyrenaica, species three previously reported ploidy levels, namely diploid (2n = 2x = 22), tetraploid (2n = 4 x = 44) and most likely very rare triploid (2n = 3x = 33). The present study aims at geographical pattern of ploidal diversity in central Europe, genetic variation and selected biological features of this species. Flow cytometru, analysis of two chloroplast DNA markers (trnF-trnL and cp018-cp020), morphometrics and in vitro germination experiments were used. The tetraploid cytotype occurs throughout the studied area; in the Czech Republic it was found in the České středohoří Mts., southern Bohemia and Moravia, in Slovakia it especially in the Carpathian mountains in western and central parts of the country and in the Pieniny Mts. in the northeast. Tetraploid cytotype occurs mainly in central-east Europe; in the Czech Republic it is common in the lowlands along the river of Labe and locally in Moravia; in Slovakia it was found along the river of Váh, in the Slovenské rudohorie Mts. and in the Belanské Tatry Mts. A mixed-ploidy population (2x, 4x) was discovered in southern...