Fundação de Amparo a Pesquisa do Estado de Minas Gerais / The of more than an embryo per seed, known as polyembryony, and the asexual reproduction via seed, known as apomixis, are usually associated. The records and importance of these events have increased in the last decades both for the Angiosperms as a whole and for specific biomes, as the Cerrado, where sexual reproduction is predominant. Despite these records, information on the ecology of polyembryonic seeds is still lacking. In this scenario, we aimed to confirm polyembryony as an indicator of apomixes in woody species of Cerrado and, based on studies of embryo resource allocation, seed germination, emergency and seedling growth, try to define the advantages and disadvantages of polyembryony for the ecology of these species. Out of a 132 species sample, 30.30% presented polyembryony, although only 6.82% did have the percentage of polyembryonic seeds above 5% and were more readily associated with apomixes. Among the Melastomataceae, a characteristic Neotropical family very well represented in the Cerrado, 33.96% of the species presented polyembryony, which predominated in the tribe Miconieae. Both in the Cerrado as a whole and in the Melastomataceae in particular, the polyembryony was associated with apomixes, but in the later, the apomixes was less often associated to higher frequencies of polyembryony. More specific studies with species of Eriotheca (Malvaceae-Bombacoideae) showed mosaics of monoembryonic and polyembryonic populations in E. gracilipes and E. pubescens, with ecological differences between either types of population. Polyembryony did not affected markedly seed germinability and seedling emergence, but resulted in less homogeneous processes than the one found in monoembryonic seeds. Resource allocation studies showed that both embryo and seedling mass decreased with the number of embryos or seedling per seed. These differences reduced the survival ability of seedlings emerging from polyembryonic seeds but, on the other hand, increased the survival chances of at least one of the seedlings from these seeds (seed individual survival). In Handroanthus chrysotrichus (Bignoniaceae), some of the putative advantages of polyembryony, as allee effect and bet hedging could be experimentally evaluated. Polyembryony was clearly correlated with the occurrence of apomixis and the results presented here showed that both processes can be important factors for the persistence and distribution of plant species in the Cerrado Biome. / A ocorrência de mais de um embrião por semente, conhecida como poliembrionia, e a reprodução assexuada via semente, conhecida como apomixia, são processos comumente associados. Os registros e a importância destes eventos vêm aumentando nas últimas décadas, tanto entre as Angiospermas como um todo quanto em biomas como o Cerrado, reconhecido como um bioma onde a reprodução sexuada é predominante. Apesar destes registros, existem ainda poucas informações sobre a ecologia das sementes poliembriônicas. Nesse cenário, objetivou-se confirmar a poliembrionia como indicador de apomixia em espécies arbóreas e arbustivas de Cerrado e, a partir de estudos alocação de recursos entre embriões, de germinação de sementes, emergência e crescimento de plântulas, tentar definir quais são as vantagens e desvantagens da poliembrionia para a ecologia das espécies. De um total de 132 espécies amostradas, 30,30% apresentaram poliembrionia, embora somente 6,82% apresentem porcentagens de sementes poliembriônicas acima de 5% e sejam mais diretamente associadas a presença de apomixia. Entre as melastomatáceas, uma família característica dos Neotrópicos e muito bem representada no Cerrado, 33,96% das espécies apresentam poliembrionia, com predominância em espécies da tribo Miconieae. Nos dois grupos amostrados, a poliembrionia foi associada à espécies apomíticas, poliplóides e com amplo padrão de distribuição, mas entre as Melastomataceae, a apomixia nem sempre estava associada a freqüências altas de poliembrionia. Estudos mais específicos com espécies de Eriotheca (Malvaceae-Bombacoideae) mostraram a ocorrência de mosaicos de populações monoembriônicas e poliembriônicas em E. gracilipes e E. pubescens, com diferenças na ecologia dos diferentes tipos de populações. A poliembrionia afetou pouco a germinabilidade e emergência das sementes, mas produziu um processo menos homogêneo que aquele das sementes monoembriônicas. Os estudos de alocação mostraram que a massa dos embriões diminuiu com o número de embriões por semente, fator que se repetiu com a massa individual de cada plântula. Estas diferenças de alocação reduziram a capacidade de sobrevivência das plântulas de sementes poliembriônicas, mas de uma maneira geral, aumentaram as possibilidades de sobrevivência de alguma das plântulas destas sementes. Em Handroanthus chrysotrichus (Bignoniaceae) algumas das vantagens da poliembrionia como a ocorrência de allee effect e bet hedging puderam ser avaliadas experimentalmente. A poliembrionia foi fortemente correlacionada com a presença de apomixia e os resultados mostram que a presença de poliembrionia e apomixia podem ser fatores importantes para a persistência e distribuição de espécies no Bioma Cerrado. / Doutor em Ecologia e Conservação de Recursos Naturais

Environmental conditions can influence epigenetic variability of clonal organisms. These environmentally induced changes have potential to persist in following generations. In theory, this transgenerational memory could allow clonal (and especially apomictic) plants to partly compensate reduced genetic variability and could enhance adaptative ability of apomictic populations. According to Muller's ratchet model, asexual organisms amass deleterious mutations, which could lead to their extinction. Possible transgenerational memory could partly compensate for this. It could also account for longtime survivorship of asexual organisms in environment. This thesis deals with transgenerational memory of triploid apomictic populations of Hieracium alpinum, with completely missing sexuality and thus genetically uniform offspring. Transgenerational effect in Hieracium alpinum was studied on clonal lineages originated from 5 populations (Norway, Austria, Bosnia and Herzegovina and Slovakia). Seeds were planted in cultivation experiment I in 3 treatments (control, added nutrients, added salicylic acid) and vegetative and generative traits were measured. Plants responded to nutrient addition but not to salicylic acid. Treatment with nutrient addition increased biomass, number of leaves and SLA (specific leaf...

The mode of reproduction can greatly influence the demography and the evolutionary success of the taxon. In the case of autonomous asexual formation seeds are apomictic taxa fully independent of pollinators and compatible partners. For sexual taxa with strict autoincompatibility it is the opposite, i.e. sexual taxa need pollinators and compatible partners for birth of offspring. Therefore, in marginal population and for more extreme situation with lower pollinating activity will have apomictic taxa a higher level of reproductive assurance than sexual taxa vascular plants. This hypothesis was tested in natural populations of apomictic and sexual taxa. In the diploma thesis we therefore investigate the method mode of reproduction and reproductive assurance of 52 taxa of the genus Hieracium s. str. (family Asteraceae) in Europe. Of these, 12 were diploid sexually diploid taxa and 42 polyploid apomictic reproductive taxa. From these taxa we harvested seeds from fully developed capitulum and we determined the potential (total number of seeds in the capitulum) and the realized (the percentage of well-developed seeds at the capitulum). The ploidy of the offspring (the embryos and the seedling) and method origins of seeds we examined using flow cytometry. The results show that the plants of diploid species...

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Apomixis is an asexual mechanism of reproduction through seed formation with clonal embryos. The high rate of polyembryonic seeds is usually an indicative for the occurrence of sporophytic apomixis and in Bignoniaceae the expression of sporophytic apomixis seems to be related to polyploidy. Handroanthus ochraceus and H. chrysotrichus are two tree species of Bignoniaceae that area morphologically related, which usually leads to identification problems. Handroanthus ochraceus has self-incompatible non-apomictic and monoembryonic populations and self-fertile sporophytic apomictic and polyembryonic populations. H. chrysotrichus has only self-fertile, sporophytic apomictic and polyembryonic populations recognized until now. To understand the genetic and phenotypic consequences of this reproductive mosaic in these populations it were conducted molecular and morphological analysis in the present study. The aims of molecular analyzes were to verify the genetic variability of H. ochraceus populations, compare the genetic diversity parameters investigating the propose of a possible relationship between the reproductive systems and genetic diversity in this species and still understand the possible relationships between apomictic and non-apomictic populations. The ten primers resulted in 104 fragments and the genetic diversity parameters showed a high mean for the populations (P = 66.35%, I = 0.341 and He = 0.227). In this study no clones were detected. The AMOVA analysis showed the higher variation within populations (61%), like allogamous populations. Cluster analysis and Bayesian analysis of genetic assignment determined two distinct groups, one consisting of two non-apomictic populations, Pires do Rio and Biribiri, and other gathering the apomictic populations and the non-apomictic from Uberlândia. The strong relationship between the self-incompatible population of Uberlândia and the apomictic populations indicates separate origins for non-apomictic populations of H. ochraceus. This relationship between self-incompatible population of Uberlândia and self-fertile populations suggest that these non-apomictic individuals may have the same genetic constitution of a possible ancestral population of self-fertile populations. The morphological analysis aimed to find differences between monoembryonic and polyembryonic populations of H. and morphological traits to separate the H. chrysotrichus and H. ochraceus. With this purpose non-apomictic monoembryonic populations and apomictic polyembryonic populations of H. ochraceus and an apomictic polyembryonic population of H. chrysotrichus were analyzed. Qualitative and quantitative morphological characteristics of leaf, flower and fruit were evaluated through univariate and multivariate analyzes. The pollen grain area, the length of peduncles, the width of stigma and the number of extrafloral nectaries in the calyx showed higher values for polyembryonic populations of H. ochraceus so they could be an evidence of polyploidy in H. ochraceus, since these larger measures may be associated with the cell volume increase usually entailed by polyploidy. Several morphological characters separated H. ochraceus from H. chrysotrichus, which can support the identification of individuals and the distinction between the two species. Therefore, the morphometric analysis determined that morphological features can make a distinction between mono and polyembryonic populations of H. ochraceus and the association of this methodology with reproductive biology analysis can be a complement for the determination of ploidies and reproductive system in both species. / A apomixia é um mecanismo de reprodução assexuada através da formação de sementes com embriões clonais. A elevada taxa de sementes poliembriônicas é um indicativo da ocorrência da apomixia esporofítica e, em Bignoniaceae a expressão da apomixia esporofítica parece estar vinculada à poliploidia. Handroanthus ochraceus e H. chrysotrichus são duas espécies arbóreas de Bignoniaceae morfologicamente semelhantes, o que normalmente ocasiona problemas de identificação. Handroanthus ochraceus apresenta populações autoincompatíveis, não apomíticas e monoembriônicas, e populações autoférteis, apomíticas esporofíticas e poliembriônicas. Já H. chrysotrichus apresenta apenas populações autoférteis, apomíticas esporofíticas e poliembriônicas conhecidas. Para compreender as consequências genéticas e fenotípicas deste mosaico reprodutivo foram realizadas análises moleculares e morfológicas. As análises moleculares tiveram por objetivos determinar a variabilidade genética de populações de H. ochraceus, comparar os parâmetros de diversidade genética, averiguar a possível relação entre os sistemas reprodutivos e a diversidade genética na espécie e ainda compreender as possíveis relações entre populações apomíticas e não apomíticas. Os dez primers ISSR utilizados resultaram em 104 bandas e os parâmetros de diversidade genética apresentaram uma média elevada para as populações (P = 66,35%, I = 0,341 e He = 0,227). Não foram detectados clones nas amostras populacionais. O cálculo da AMOVA demonstrou alta variação genética dentro das populações (61%), semelhante a populações alógamas. A análise de agrupamento e a análise Bayesiana de atribuição genética determinaram a formação de dois grupos distintos, um constituído por duas populações não apomíticas, Pires do Rio e Biribiri, e o outro reunindo as populações apomíticas e a não apomítica de Uberlândia. A forte relação entre a população autoincompatível de Uberlândia e as apomíticas indica origens distintas para as populações não apomíticas de H. ochraceus. Esta relação entre a população autoincompatível de Uberlândia e as populações autoférteis permite supor que os indivíduos não apomíticos dessa população apresentem a mesma constituição genética de uma possível população ancestral das populações autoférteis. As análises morfológicas tiveram por objetivo buscar diferenças entre populações mono e poliembriônicas de H. ochraceus e determinar características morfológicas que auxiliam na separação de H. ochraceus e H. chrysotrichus, para isso populações reconhecidamente monoembriônicas não apomíticas e poliembriônicas apomíticas de H. ochraceus e uma população poliembriônica apomítica de H. chrysotrichus foram analisadas. Características qualitativas e quantitativas de folha, flor e fruto foram avaliadas utilizando análises estatísticas univariadas e multivariadas. As características área do grão de pólen, comprimento do pedicelo, largura do estigma e número de nectários extraflorais no cálice apresentaram maiores valores para populações poliembriônicas de H. ochraceus e poderiam ser uma evidencia de poliploidia em H. ochraceus, pois, podem estar associadas ao aumento do volume celular geralmente acarretado pela poliploidia. Diferentes características morfológicas separaram as populações de H. ochraceus da população de H. chrysotrichus, as quais podem auxiliar na identificação de indivíduos e na distinção das espécies. Assim as análises morfométricas determinaram que características morfológicas de alguma forma podem distinguir populações mono e poliembriônicas de H. ochraceus, e em conjunto com análises de biologia reprodutiva podem ser uma metodologia auxiliar na determinação da ploidia e sistema reprodutivo. / Mestre em Genética e Bioquímica

Les systèmes reproducteurs déterminent comment le matériel génétique est transmis d’une génération à la suivante […]. Les espèces qui combinent de la reproduction sexuée et asexuée/clonale sont très répandues [… mais] l’effet de leur système reproducteur sur leur évolution reste énigmatique et discuté.L’objectif de cette thèse est de modéliser la dynamique des fréquences génotypiques d’une population avec une combinaison de reproduction sexuée et/ou clonale dans des cycles de vie principalement diploïdes [. … Un] modèle du type chaine de Markov avec temps et états discrets sert de base mathématique pour décrire [leurs] changements […] au cours du temps.Les résultats montrent que la reproduction partiellement asexuée peut en effet modifier la dynamique de la diversité génomique par rapport à une reproduction strictement sexuée ou strictement asexuée. […] L’histoire démographique a un rôle important pour les organismes partiellement clonaux et doit être prise en compte dans toute analyse […].Cette thèse fait des recommandations pour la collecte des données et une hypothèse de base pour l’interprétation des données de génétique/génomique […]. Ces résultats ont des retombées dans plusieurs domaines, allant de la recherche fondamentale […] à des applications en agriculture […], pêche […] et protection de la nature […]. / Reproductive systems determine how genetic material is passed from one generation to the next, making them an important factor for evolution. Organisms that combine sexual and asexual/clonal reproduction are very widespread [… yet] the effects of their reproductive system on their evolution are still controversial and poorly understood.The aim of this thesis was to model the dynamics of genotype frequencies under combined sexual/clonal reproduction in dominantly diploid life cycles [. … A] state and time discrete Markov chain model served as the mathematical basis to describe [their] changes […] through time.The results demonstrate that partial clonality may indeed change the dynamics of genomic diversity compared to either exclusively sexual or exclusively clonal populations. […] Time has a crucial role in partially clonal populations and needs to be taken into account in any analysis of their genomic diversity.This thesis provides recommendations for data collection and a null hypothesis for the interpretation of population genetic/genomic data […]. Moreover, it includes new methods for the analysis of genotype-based population genetic Markov chain models. These results have a high potential relevance in several areas, ranging from basic research […] to applications in agriculture […], fisheries […] and nature conservation […].

This thesis is a biosystematic study focusing on the taxonomy and variability of selected taxa of the genus Sorbus, one of the most diverse and taxonomically complicated plant groups in Europe. Classical and modern biosystematic methods - comparative study, chromosome counting, analysis of nuclear microsatellite markers, flow cytometry, and traditional multivariate and outline morphometric analyses were used to assess the morphological, karyological and genetic variability of the genus. The final synthesis of these approaches led to the description of several new taxa (species, hybrids and a subgenus) and the correction of several taxonomic misinterpretations. To reveal ongoing evolutionary processes responsible for the generation of the observed variability, the reproductive modes of 42 Sorbus taxa were examined using flow-cytometric seed and pollen screens. Apart from revealing major trends, the study estimates the frequency of rare events and provides several novel conclusions that are relevant both specifically to Sorbus/Rosaceae and to apomixis in general.