<p>  </p> <p>Tar spot is a new foliar disease of corn in the United States. Tar spot was first detected in 2015 and is now among the most important corn diseases in the Midwest. Tar spot is caused by the obligate biotrophic fungus, <em>Phyllachora maydis</em> Maubl, from the genus, <em>Phyllachora </em>which consist of over 1,200 species of host-specific fungi. Due to the recent emergence, studies relating to <em>P. maydis</em> population dynamics in the U.S. are limited. How much genetic diversity, variation, and level of gene flow are occurring within and among these populations? Knowledge of the population dynamics is imperative for understanding the pathogen’s biology, ecology, epidemiology, and management. Currently, no corn hybrid is fully resistant to tar spot. Foliar fungicides are currently the most effective option for disease management, but best practices for fungicide management remain unknown. Better information is needed on fungicide efficacy and fungicide application timing to reduce tar spot severity, protect yield, and increase profitability for Indiana corn growers. </p> <p>This research dissertation presents four chapters to answers those questions and bridge the gaps between the knowns and unknows of this novel corn-<em>Phyllachora maydis</em> pathosystem. <strong>Chapter 1</strong> presents a literature review on tar spot of corn, its economic impact, the causal pathogen, its host, lifecycle, distribution, and known management strategies as a resource for understanding the pathosystem in the U.S. <strong>Chapter 2</strong> examines the genetic population structure, diversity, geneflow and mode of reproduction in Midwest U.S. by employing microsatellite (SSR) markers. <strong>Chapter 3</strong> presents results from multi-year, multi-location, small-plot field trials on the net return of foliar fungicides and fungicide timing on tar spot management in Indiana. Lastly, <strong>Chapter 4</strong> concludes by evaluating of an integrated management strategy for tar spot by examining the integration of tillage, corn hybrids and fungicide application in reducing tar spot severity while protecting yields.</p> <p>Results provided in this research dissertation will be used to guide future studies and provide stakeholders such as researchers, corn growers, extension personnel in academia and industry with valuable information needed to guide effective disease management decisions. </p>

Genealogy reconstruction is widely used in biology when relationships among entities are studied. Phylogenies, or evolutionary trees, show the differences between species. They are of profound importance because they help to obtain better understandings of evolutionary processes. Pedigrees, or family trees, on the other hand visualize the relatedness between individuals in a population. The reconstruction of pedigrees and the inference of parentage in general is now a cornerstone in molecular ecology. Applications include the direct infer- ence of gene flow, estimation of the effective population size and parameters describing the population’s mating behaviour such as rates of inbreeding. In the first part of this thesis, we construct genealogies of various types of cancer. Histopatho- logical classification of human tumors relies in part on the degree of differentiation of the tumor sample. To date, there is no objective systematic method to categorize tumor subtypes by maturation. We introduce a novel algorithm to rank tumor subtypes according to the dis- similarity of their gene expression from that of stem cells and fully differentiated tissue, and thereby construct a phylogenetic tree of cancer. We validate our methodology with expression data of leukemia and liposarcoma subtypes and then apply it to a broader group of sarcomas and of breast cancer subtypes. This ranking of tumor subtypes resulting from the application of our methodology allows the identification of genes correlated with differentiation and may help to identify novel therapeutic targets. Our algorithm represents the first phylogeny-based tool to analyze the differentiation status of human tumors. In contrast to asexually reproducing cancer cell populations, pedigrees of sexually reproduc- ing populations cannot be represented by phylogenetic trees. Pedigrees are directed acyclic graphs (DAGs) and therefore resemble more phylogenetic networks where reticulate events are indicated by vertices with two incoming arcs. We present a software package for pedigree reconstruction in natural populations using co-dominant genomic markers such as microsatel- lites and single nucleotide polymorphism (SNPs) in the second part of the thesis. If available, the algorithm makes use of prior information such as known relationships (sub-pedigrees) or the age and sex of individuals. Statistical confidence is estimated by Markov chain Monte Carlo (MCMC) sampling. The accuracy of the algorithm is demonstrated for simulated data as well as an empirical data set with known pedigree. The parentage inference is robust even in the presence of genotyping errors. We further demonstrate the accuracy of the algorithm on simulated clonal populations. We show that the joint estimation of parameters of inter- est such as the rate of self-fertilization or clonality is possible with high accuracy even with marker panels of moderate power. Classical methods can only assign a very limited number of statistically significant parentages in this case and would therefore fail. The method is implemented in a fast and easy to use open source software that scales to large datasets with many thousand individuals.:Abstract v Acknowledgments vii 1 Introduction 1 2 Cancer Phylogenies 7 2.1 Introduction..................................... 7 2.2 Background..................................... 9 2.2.1 PhylogeneticTrees............................. 9 2.2.2 Microarrays................................. 10 2.3 Methods....................................... 11 2.3.1 Datasetcompilation ............................ 11 2.3.2 Statistical Methods and Analysis..................... 13 2.3.3 Comparison of our methodology to other methods . . . . . . . . . . . 15 2.4 Results........................................ 16 2.4.1 Phylogenetic tree reconstruction method. . . . . . . . . . . . . . . . . 16 2.4.2 Comparison of tree reconstruction methods to other algorithms . . . . 28 2.4.3 Systematic analysis of methods and parameters . . . . . . . . . . . . . 30 2.5 Discussion...................................... 32 3 Wild Pedigrees 35 3.1 Introduction..................................... 35 3.2 The molecular ecologist’s tools of the trade ................... 36 3.2.1 3.2.2 3.2.3 3.2.1 Sibship inference and parental reconstruction . . . . . . . . . . . . . . 37 3.2.2 Parentage and paternity inference .................... 39 3.2.3 Multigenerational pedigree reconstruction . . . . . . . . . . . . . . . . 40 3.3 Background..................................... 40 3.3.1 Pedigrees .................................. 40 3.3.2 Genotypes.................................. 41 3.3.3 Mendelian segregation probability .................... 41 3.3.4 LOD Scores................................. 43 3.3.5 Genotyping Errors ............................. 43 3.3.6 IBD coefficients............................... 45 3.3.7 Bayesian MCMC.............................. 46 3.4 Methods....................................... 47 3.4.1 Likelihood Model.............................. 47 3.4.2 Efficient Likelihood Calculation...................... 49 3.4.3 Maximum Likelihood Pedigree ...................... 51 3.4.4 Full siblings................................. 52 3.4.5 Algorithm.................................. 53 3.4.6 Missing Values ............................... 56 3.4.7 Allelefrequencies.............................. 58 3.4.8 Rates of Self-fertilization.......................... 60 3.4.9 Rates of Clonality ............................. 60 3.5 Results........................................ 61 3.5.1 Real Microsatellite Data.......................... 61 3.5.2 Simulated Human Population....................... 62 3.5.3 SimulatedClonalPlantPopulation.................... 64 3.6 Discussion...................................... 71 4 Conclusions 77 A FRANz 79 A.1 Availability ..................................... 79 A.2 Input files...................................... 79 A.2.1 Maininputfile ............................... 79 A.2.2 Knownrelationships ............................ 80 A.2.3 Allele frequencies.............................. 81 A.2.4 Sampling locations............................. 82 A.3 Output files..................................... 83 A.4 Web 2.0 Interface.................................. 86 List of Figures 87 List of Tables 88 List Abbreviations 90 Bibliography 92 Curriculum Vitae I

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Cette thèse est consacrée à la reconnaissance individuelle et au choix du partenaire chez la mouette tridactyle (Rissa tridactyla), espèce génétiquement monogame et fidèle à son partenaire d'une saison sur l'autre. Mes expériences démontrent l'existence d'une reconnaissance vocale entre les partenaires d'un couple, ainsi qu'une reconnaissance des parents par les poussins. Des observations empiriques suggèrent que les parents utilisent également le cri pour reconnaître leurs poussins. Ceci pourrait relever d'une stratégie adaptative: les poussins reconnus par leurs parents retournent plus vite au nid lors de leur premier envol, ce qui pourrait diminuer l'énergie dépensée dans des vols stressants ou des batailles avec d'autre résidents. Les analyses ont aussi montré que les distances génétiques entre membre des couples sont plus grandes qu'attendu par chance. Ceci augmente la probabilité d'hétérozygotie des poussins, et donc le succès de reproduction. En effet, le taux d'éclosion augmente avec la distance génétique entre les parents, et les poussins hétérozygotes grandissent plus vite et survivent mieux jusqu'à 25 jours. En outre, les couples formés d'individus génétiquement proches copulent moins souvent, ce qui confirmerait l'influence sur les comportements sexuels de la distance génétique intra couple. Ces résultats posent la question de la perception de la similarité génétique par les partenaires potentiels. Mes résultats préliminaires suggèrent que celle-ci ne semble pas impliquer la voix. En effet, la différence entre les cris ne semble pas corrélée à la distance génétique. Des paramètres olfactifs ou visuels pourraient donc être impliqués. Ces résultats suggèrent que le choix du partenaire pourrait donc être très important chez cette espèce génétiquement monogame.

O tucumã-do-Amazonas (Astrocaryum aculeatum), da família Arecaceae, é uma espécie com ocorrência na Amazônia Ocidental e Central brasileira. Seu fruto é muito apreciado na região amazônica que consome a polpa na forma fresca. Sua produção é feita pelos agricultores extrativistas. Não existem informações dos parâmetros de diversidade e estrutura genética das populações naturais desta espécie, fundamental para o estabelecimento de estratégias adequadas no uso deste recurso genético, e para aumentar a eficiência da domesticação e seleção para um programa de melhoramento genético. Não é conhecido também como este extrativismo pode estar influenciando a diversidade entre estas populações ao nível dos genitores e progênies, o fluxo gênico e a estrutura genética espacial. Assim, o objetivo desta pesquisa foi caracterizar a estrutura e a diversidade genética de populações de A. aculeatum no Estado do Amazonas, utilizando marcadores microssatélites. Como objetivos específicos o estudo visou desenvolver iniciadores de microssatélites para A. aculeatum; avaliar a diversidade e a estrutura genética; determinar o fluxo gênico, bem como o grau de parentesco, a estrutura genética espacial (EGE) e o coeficiente de coancestria numa população. Os iniciadores de microssatélites foram isolados a partir de uma biblioteca genômica enriquecida e caracterizados usando 40 amostras provenientes de duas populações selvagens. Foram identificados 14 iniciadores de microssatélites, mostrando um alto polimorfismo nas populações avaliadas. Para estimar a diversidade e estrutura genética foram utilizadas amostras de material vegetal de 218 plantas distribuídas em 15 populações, localizadas em 14 municípios do estado de Amazonas. Foram identificados 101 alelos nos iniciadores, com média de 10,1 alelos. As médias das heterozigosidades observadas (HO) foram superiores às esperadas (HE) ao nível de população (HO=0,639, HE=0,557) e locos (HO=0,639; HE=0,594). Os baixos valores de FST (0,07) mostraram uma moderada estrutura populacional e a análise Bayesiana indicou um agrupamento mais adequado de k=4, o que foi confirmado pelas análises de PCoA. Para realizar o fluxo gênico, EGE e coeficiente de coancestria foram utilizadas 244 amostras de material vegetal, distribuídas entre 112 possíveis genitores, 12 matrizes e 120 progênies numa população. Foi detectada EGE significativa para os adultos até a distância de 50 m, indicando dispersão de sementes a curtas distâncias. A análise de paternidade detectou 9,2% de imigração de pólen. Não foram detectadas autofecundações e a distância média de polinização dentro da população foi de 81 m. Os índices de diversidade genética confirmaram a estratégia reprodutiva por alogamia da espécie, com níveis elevados de heterozigotos nas populações, e a estrutura genética sugere que a espécie teria iniciado o processo de formação das subpopulações há pouco tempo, tendo sido influenciada antes e após o início do desmatamento. O fluxo gênico detectado indica que a entrada de alelos na população através da imigração de pólen está contribuindo para manutenção da diversidade genética da população. Com relação à conservação da espécie, esta deve ser realizada ao nível das bacias hidrográficas, direcionando políticas de manejo para as bacias hidrográficas da Amazônia. Entretanto, dentro da população é importante a renovação com novas progênies selecionadas a partir de um espaçamento mínimo de 50 m entre plantas matrizes para diminuir as probabilidades de coletar sementes de plantas parentes. / The tucum palm (Astrocaryum aculeatum), family Arecaceae, is a species occurring in Western and Central Brazilian Amazon. Its fruit is widely appreciated in the Amazon region that consumes the pulp in fresh form. Its production is made by extractive farmers. There is no information on the genetic diversity and structure of natural populations of this species, fundamental to establishing adequate strategies in the use of this genetic resource, increase the efficiency of domestication and selection for a breeding program. It is also unknown the way this extrativism may be influencing to diversity of these populations at the level of the parents and offspring, as well as gene flow and spatial genetic structure. Therefore, the objective of this research was to characterize the structure and genetic diversity of populations of A. aculeatum in Amazonas State, using microsatellite markers. As specific objectives, the study aimed to develop microsatellite primers for A. aculeatum; evaluate the genetic diversity and structure; determine the gene flow, the degree of relationship or kinship, the spatial genetic structure (SGS) and the coefficient of coancestry in one population. The loci were isolated from a microsatellite-enriched genomic library and were characterized using 40 samples coming from two wild populations. Fourteen microsatellite primers were identified, showing a high polymorphism in the populations evaluated. To estimate the genetic diversity and structure we used samples of plant material from 218 plants distributed in 15 populations located in 14 municipalities of the state of Amazonas. A total of 101 alleles were identified in the primers, with an average of 10.1 alleles. The averages of the observed heterozygosities (HO) were higher than the expected heterozygosities (HE) at the population (HO=0.639; HE=0.557) and loci (HO=0.639; HE=0.594) levels. The low values of FST (0.07) showed a moderate population structure and the Bayesian analysis indicated as the most suitable a cluster of k=4, confirmed by the groups formed in the PCoA. To conduct gene flow, SGS and the coefficient of coancestry, 244 samples of plant material were used, distributed among 112 possible progenitors, 12 matrices and 120 progenies in a population. A significant SGS was detected for adult trees up to the distance of 50 m, indicating seed dispersal over short distances. Paternity analysis indicated 9.2% of pollen immigration. Selfing was not detected and the average distance of pollination within the population was 81 m. The genetic diversity indices confirmed the outcrossing reproductive strategy of the species with high levels of heterozygotes in the populations, and the genetic structure suggests that the species would have started the process of formation of subpopulations a short time ago, being influenced before and after the start of deforestation. The gene flow detected indicates that the entry of alleles in the population through pollen immigration is contributing to the maintenance of genetic diversity of the population. With respect to the conservation of the species, it should be performed at the level of the river basins, directing management policies for the river basins of the Amazon. However, within the population it is important to renew with new progenies selected from a minimum spacing of 50 m between plant matrices to decrease the probability of collecting seeds from plant relatives.

Leishmania infantum ist für einen beträchtlichen Teil der viszeralen Leishmaniosen weltweit verantwortlich. Die allgemein anerkannte Methode zu Klassifizierung wie auch zur Identifizierung von Leishmanien ist die Isoenzymanalyse. Speziell für L. infantum reicht das Auflösungsvermögen dieser Methodik allerdings nicht aus, da mehr als 80 % der analysierten Stämme dem prädominanten Zymodem MON-1 angehören. In dieser Arbeit wurden Primerpaare für die Amplifikation von 15 unabhängigen Mikrosatelliten entwickelt. Das verwendete Protokoll zur Erstellung einer angereicherten genetischen Bibliothek ist leicht für andere Leishmanienspezies adaptierbar. Die Marker wurden an 13 Stämmen des L. donovani-Komplexes getestet mit Schwerpunkt auf L. infantum MON-1. Die berechneten Dendrogramme entsprachen weitestgehend den Ergebnissen früherer, auf genotypischen Methoden basierenden phylogenetischen Studien. In Abhängigkeit von Zymodem und Spezies wurden unterschiedlich hohe Anteile heterogener Allele beobachtet. Das ist der erste Satz von unabhängigen Mikrosatellitenmarkern entwickelt speziell für L. infantum, der groß genug für phylogenetische Anwendungen ist. Für jedem untersuchten Stamm wurde ein eigener Multilocus-Genotyp beobachtet, was diese Methode zu einem wichtigen Werkzeug für epidemiologische und populationsgenetische Untersuchungen macht. / Leishmania infantum is responsible for a large proportion of visceral leishmaniasis all over the world. The universally accepted standard method for characterizing and identification of Leishmania is the isoenzyme analysis. Especially for L. infantum, this procedure lacks of discriminative power because the predominant zymodeme MON-1 is represented by more than 80 % of the analysed isolates. In this study, PCR assays amplifying 15 independent microsatellites were developed using a method to create highly enriched libraries that can easily be adapted for other species of Leishmania. The panel of markers was tested for 13 strains of the L. donovani complex with main emphasis on L. infantum MON-1. The calculated dendrograms corresponded to the results of former phylogenetic investigations based on genotypic methods. Depending of zymodeme and species, different degrees of allelic heterozygosity were observed. This is the first set of independent microsatellite markers especially developed for L. infantum, which is large enough for phylogenetic applications. An unique multi locus genotype is produced for each analysed isolate what makes this approach to a powerful tool for epidemiological and population genetic investigations.