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...

A lot of the research has been made in the field of plant polyploidy since the discovery of this phenomenon. However, the more we know, the more questions arise. Overall, the most insistent questions remain: How did the polyploids arise and become established? Is it advantageous to be polyploid? We chose a perennial herb Vicia cracca L. to study the causes and consequences of polyploidisation on microevolution of a diploid-tetraploid complex in central European conditions. First, evidence from both flow cytometry and molecular markers (allozymes, DNA sequences, microsatellites) confirmed a hypothesised autopolyploid origin of tetraploids. Based on allozymes, we proved that tetraploids are genetically richer than diploid ancestors. However, we showed that the conclusions depend on statistics used for genetic variation measurements. Nevertheless, tetraploids of V. cracca suffered lesser reduction in seed production after artificial selfing than diploids. We thus infer that they profit from multiple allele dosage, which can mask deleterious alleles. We also corroborated an existence of a central European contact zone of diploids and tetraploids and discovered two other contact zones in south-western and south- eastern Europe. The central European contact zone with several mixed-ploidy populations served us as...

A lot of the research has been made in the field of plant polyploidy since the discovery of this phenomenon. However, the more we know, the more questions arise. Overall, the most insistent questions remain: How did the polyploids arise and become established? Is it advantageous to be polyploid? We chose a perennial herb Vicia cracca L. to study the causes and consequences of polyploidisation on microevolution of a diploid-tetraploid complex in central European conditions. First, evidence from both flow cytometry and molecular markers (allozymes, DNA sequences, microsatellites) confirmed a hypothesised autopolyploid origin of tetraploids. Based on allozymes, we proved that tetraploids are genetically richer than diploid ancestors. However, we showed that the conclusions depend on statistics used for genetic variation measurements. Nevertheless, tetraploids of V. cracca suffered lesser reduction in seed production after artificial selfing than diploids. We thus infer that they profit from multiple allele dosage, which can mask deleterious alleles. We also corroborated an existence of a central European contact zone of diploids and tetraploids and discovered two other contact zones in south-western and south- eastern Europe. The central European contact zone with several mixed-ploidy populations served us as...

Gliomas are brain tumors arising from neuroglia. In most cases astrocytic or oligodendroglial component is the main element of the tumor. Non-random chromosomal abberations are found in tumor cells as was revealed previously. The aim of this study was a fluorescence in-situ hybridisation analysis (FISH) of tissue samples obtained during neurosurgical procedures, determine the frequence of selected chromosomal abberations, further correlation with morphological and clinical data and statistical analysis of the results. During six years 264 tissue samples were gained in which FISH with defined probes was performed. The acquired results were compared with histological analysis and selected clinical data (age, Karnofsky score, extent of resection, overall survival). The whole series was divided into 7 groups by tumor type for further statistical analysis. In every group median and mean survival time was calculated, Kaplan-Meier analysis was focused on influence of selected parameters to overall survival. In some categories Cox regression model was created to achieve a hazard ratio of selected parameters. In WHO Grade II and III tumors the risk of malignant progression and tumor upgrading is significantly higher in comparison with samples where specific abberations were not found (EGFR amplification, CDKN2A and...

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.

Gliomas are brain tumors arising from neuroglia. In most cases astrocytic or oligodendroglial component is the main element of the tumor. Non-random chromosomal abberations are found in tumor cells as was revealed previously. The aim of this study was a fluorescence in-situ hybridisation analysis (FISH) of tissue samples obtained during neurosurgical procedures, determine the frequence of selected chromosomal abberations, further correlation with morphological and clinical data and statistical analysis of the results. During six years 264 tissue samples were gained in which FISH with defined probes was performed. The acquired results were compared with histological analysis and selected clinical data (age, Karnofsky score, extent of resection, overall survival). The whole series was divided into 7 groups by tumor type for further statistical analysis. In every group median and mean survival time was calculated, Kaplan-Meier analysis was focused on influence of selected parameters to overall survival. In some categories Cox regression model was created to achieve a hazard ratio of selected parameters. In WHO Grade II and III tumors the risk of malignant progression and tumor upgrading is significantly higher in comparison with samples where specific abberations were not found (EGFR amplification, CDKN2A and...

Raps wurde als Fallstudie für eine Kulturpflanze ausgewählt, die sich außerhalb des Anbaus spontan etablieren kann und so als Quelle für Neophyten dient. In einer regionalen Studie im Osnabrücker Land wurde basierend auf Kartierungen (2004-2008) und kernkodierten Mikrosatellitenmarkern die Herkunft, Persistenz und genetische Variation verwilderter Rapspopulationen untersucht. Die wichtigsten Quellen für die erhöhte genetische Variation in verwilderten Rapspopulationen waren die mehrfache Einschleppung verschiedener Sorten und Hybridisierungen zwischen diesen. Raps tritt im Osnabrücker Land mit einer Reihe nah verwandter Arten, darunter Brassica rapa und Raphanus raphanistrum, sympatrisch in verwilderten Populationen auf. Hybride zwischen Raps und diploiden Rübsen konnten nachgewiesen werden. Tetraploider Rübsen wurde als Kreuzungspartner von Raps erstmals in dieser Studie untersucht. In Kreuzungsversuchen und in einem Freilandversuch konnten interspezifische Hybridisierungen zwischen tetraploiden Rübsen und Raps nachgewiesen werden. Unter natürlichen Bestäubungsbedingungen im Freilandversuch mit tetraploiden Rübsen-Mutterpflanzen wurde eine Hybridisierungsrate von 16,2% abgeschätzt. Die Untersuchungen zeigen, dass verwilderte Rapspopulationen durchaus ein Potenzial zu evolutiven Entwicklungen besitzen. Verwilderte Rapspopulationen können als Trittsteine für intra- und interspezifischen Genfluss dienen und dadurch beim Anbau transgener Rapssorten die Koexistenz verschiedener Anbauformen erschweren. Sie können die Etablierung von Transgenen außerhalb des Anbaus ermöglichen, die Transgenausbreitung fördern und Introgression in nah verwandte Arten vermitteln. Die Wahrscheinlichkeit von Hybridisierungen zwischen Raps und Rübsen (diploid und tetraploid) könnte verringert werden, wenn vermieden würde, diese kreuzkompatiblen Arten gemeinsam in Saatgutmischungen zu verwenden.

Wachstumsfaktor bedingte Aktivierung des PI3K/Akt/mTOR Signalweg wirkt positiv auf Vermehrung und Überleben. Konstitutive Aktivierung des Signalweges in NHL ist jedoch an Tumorprogression und Therapieresistenz beteiligt. Am Zelllinienmodell wurden zwei mögliche Therapiestrategien der PI3K/Akt/mTOR Inhibition erprobt, PI3K Inhibition mit BKM120 und horizontale Kombination von Zytostatika mit PI3K/Akt/mTOR Inhibitoren Erstens, BKM120 hat Antitumoraktivität in NHL und induziert Zelltod. Auf molekularer Ebene führt BKM120 vermittelte Dephosphorylierung von CDK1 an Y15 zur Aktivierung des M-Phase Komplex CDK1/Zyklin B und Eintritt in die Mitose. Parallel erlaubt die Degradation von Zyklin A und Hochregulation von Zyklin B Progression bis zur Metaphase, hemmt jedoch die Transition in die Anaphase. Anhaltender Metaphasearrest bewirkt programmierten Zelltod über den intrinsischen Signalweg der Apoptose durch Hochregulation der BH3-onlys Puma und Hrk, Aktivierung von Bax/Bak und proteolytische Spaltung von Caspase 9. Verlust von Bax/Bak oder Caspase Inhibition schützt vor BKM120 vermitteltem Zelltod. Bax/Bak defiziente Zellen, welche zusätzlich p53 Mutationen aufweisen, werden polyploid. Die Polyploidie ist ATM-MEK1/2 abhängig und kann mit Caffeine oder U0126blockiert werden. Zur Vermeidung von Polyploidie bedingter Tumorprogression, sollte BKM120 nur in Verbindung mit MAPK/ATM Inhibitoren verwendet werden. Zweitens. Horizontale Kombination PI3K/Akt/mTOR Inhibitoren mit cytotoxischen Substanzen schützt vor Apoptose. Der Schutzeffekt tritt auschließlich bei niedrigen Konzentration auf und ist unabhängig von der Art des Inhibitors bzw. Ebene der Inhibition. Das Onkogen und NFkB Target Pim-2 ist möglicherweise am Schutzmechanismus beteiligt. Durch die PI3K/Akt/mTOR vermittelte Pim-2 Regulation ergibt sich eine neue Rückkopplungsschleife. Im Fazit erschwert die Komplexizität des PI3K/Akt/mTOR Signalweges die Etablierung von Therapien. / Growth factor mediated activation of the PI3K/Akt/mTOR pathway positively regulates proliferation and survival. Constitutive activation in NHL, however, is correlated with tumor progression and therapeutic resistance. Therefore, two possible strategies were tested in a cell line model system, Inhibition of PI3K with BKM120 and PI3K/Akt/mTOR Inhibition in addition to cytostatic drug administration. First, it is demonstrated that the pan PI3K inhibitor BKM120 has antitumor activity in NHL and induces cell death. On molecular level, BKM120 mediated dephosphorylation of CDK1 on Y15 causes activation of the M-phase complex CDK1/Cyclin B and entry into mitosis. In parallel, degradation of Cyclin A and Upregulation of Cyclin B enables progression into metaphase but inhibits transition into anaphase. Prolonged metaphase arrest induces programmed cell death via the intrinsic apoptosis pathway by upregulation of the BH3-onlys Puma and Hrk, activation of Bax/Bak and proteolytic cleavage of caspase-9. Loss of Bax/Bak or caspase inhibition protects from BKM120 induced apoptosis. Bax/Bak deficient cells with additional p53 mutation become polyploid. This polyploidy is ATM-MEK1/2 dependent and can be blocked with Caffeine or U0126. To prevent polyploidy related tumor progression, BKM120 should administered only in combination with ATM or MEK inhibitors. Second, combination of PI3K/Akt/mTOR inhibitors with cytotoxic agents protects from apoptosis. The protective effect is only detectable with low PI3K/Akt/mTOR inhibitor concentrations and independent of inhibitor type or cascade level. The oncogene and NFkB target is possibly involved in apoptosis protection and inhibition of NFkB neutralizes the protective effect. PI3K/Akt/mTOR mediated Pim-2 regulation reveals a new feedback loop within the pathway. In conclusion, the complexity of the PI3K/Akt/mTOR pathway impedes therapeutic targeting.

The thesis aims at clarifying the taxonomic difficulties in the Tortula muralis complex. For the first time, the group was studied by means of morphometric analysis, ploidy level assessment by flow cytometry, and molecular analysis based on sequencing of ITS region of nuclear ribosomal DNA. Morphometric and cytometric studies found only two well distinguished groups within the complex. These groups corresponded to the markedly variable species T. muralis and to the rather uniform species T. lingulata. Variability in ITS sequences suggested extensive gene flow among some of traditional morphologically defined taxa of the complex. Multiple polytopic autopolyploid origin of polyploids was revealed in some taxa. Changes in taxonomic conception of T. muralis complex were proposed in order to reflect structure of morphological, karyological, and DNA variability in the group. Natural hybridization is probably an important evolutionary mechanism that generated morphological diversity and taxonomic complexity in the mosses of T. muralis complex.

Myriophyllum L. (watermilfoil) belongs to one of the most invasive genus of the Northern Hemisphere. The three most agressive species, one of which is native to Europe (M. spicatum), are widespread throughout the majority of United states and two of them (M. aquaticum and M. heterophyllum) are becoming invasive in several Europaean countries as well. Therefore, Europaean populations of both native and invasive species, represents ideal place where to study processes which account for watermilfoils' invasivness. In this study, flow cytometry, morphometric analyses and germination experiments were employed to evaluate cytotype and morphological variability of Myriophyllum along with its potential to hybridize and propagate generatively. Five ploidy levels were detected in Europe, however no ploidy variability has been found within any species except for M. sibiricum (hexaploids and nonaploids) and one population of M. aquaticum (hexaploids and oktoploids). Flow cytometry proved to be useful tool for purposes of distinguishing species of different ploidy level and their potential hybrids. Nevertheless, the key species - M. sibiricum and M. spicatum - among which probably even more aggressive hybrids were recently identified, have similar hexaploid level. Thus flow cytometry cannot facilitate...