The vast majority of flowering plants rely on pollinators for outcrossed reproduction. Unfortunately, many species of pollinators are in decline. Pollinators face a number of challenges, including shifting land use, climate change and pesticides. In addition, pollinator habitat may be altered or eliminated through urbanization. Fragmented urban landscapes may lengthen the distances among foraging patches, limit resources in a given patch, and reduce the availability of pollinator nesting sites. Here, I examine the effects of urbanization on pollination services by addressing the following questions: (1) Does bee species richness and/or abundance change along an urbanization gradient?; (2) Does the pollen limitation of a focal species vary along the same gradient?; (3) Do plants with a mixed mating system produce more selfed seeds in more urban environments? Using the percentage of impervious land cover in the space immediately surrounding the site as a proxy for site urbanization, 15 study sites were set up across the city of Ottawa to span a range from minimal percent impervious surface (mostly green space) to mostly impervious surface (little green space) at a range of spatial scales. At each site I set up an array of 20 potted Impatiens capensis plants as well as six pan traps in order to collect data on pollen limitation, seed production, and the number of selfed progeny, in addition to information about local pollinator species richness and abundance. Plants in the arrays were randomly assigned to either a hand or open pollination treatment in order to assess the level of pollen limitation. Surprisingly, pollinator species richness and abundance were not correlated with urbanization. Pollen limitation declined with urbanization, however, so did overall seed set, making it difficult to detangle the effects of resource limitation and hand pollination treatments. The number of selfed seeds produced by a plant was also significantly correlated with site urbanization. Further investigation is required to better understand the dynamics of pollination services in urban environments. I recommend that future studies explore how the presence of individual bee species in rural and urban habitats could be influencing pollen limitation and selfing in I. capensis.

Pollen contamination is a severe problem in production breeding programs since it reduces the expected gain. In an attempt to solve this problem Skogforsk created an isolation experiment in the advanced Scots pine (Pinus sylvestris) seed orchard Västerhus in Västerbotten, Sweden. This experiment involves covering blocks of trees with a tent during the pollination period so that they only can mate with each other inside. To evaluate the effects of this tent treatment one tree from inside a tent with supplementary mass pollination (SMP) and one tree from the open control were chosen for this study. 48 seeds from each tree were sampled and genotypes at 9 microsatellite (SSR) loci. The likelihood and exclusion methods for paternity assignment were used to establish the fathers to these seeds. The results showed 0% contamination inside the tent and 4-8% outside in the control. The number of fathers contributed to the fertilization of the 48 seeds was 9 inside and 15 outside. The selfing rate was unexpectedly high, 10% inside the tent and 19% outside. The mating system inside the tent need to be further evaluated to fully understand what other effect the treatment has on the future progeny.

Yes / Introduction The transition from cross-fertilisation (outcrossing) to self-fertilisation (selfing) frequently coincides with changes towards a floral morphology that optimises self-pollination, the selfing syndrome. Population genetic studies have reported the existence of both outcrossing and selfing populations in Arabis alpina (Brassicaceae), which is an emerging model species for studying the molecular basis of perenniality and local adaptation. It is unknown whether its selfing populations have evolved a selfing syndrome. Methods Using macro-photography, microscopy and automated cell counting, we compared floral syndromes (size, herkogamy, pollen and ovule numbers) between three outcrossing populations from the Apuan Alps and three selfing populations from the Western and Central Alps (Maritime Alps and Dolomites). In addition, we genotyped the plants for 12 microsatellite loci to confirm previous measures of diversity and inbreeding coefficients based on allozymes, and performed Bayesian clustering. Results and Discussion Plants from the three selfing populations had markedly smaller flowers, less herkogamy and lower pollen production than plants from the three outcrossing populations, whereas pistil length and ovule number have remained constant. Compared to allozymes, microsatellite variation was higher, but revealed similar patterns of low diversity and high Fis in selfing populations. Bayesian clustering revealed two clusters. The first cluster contained the three outcrossing populations from the Apuan Alps, the second contained the three selfing populations from the Maritime Alps and Dolomites. Conclusion We conclude that in comparison to three outcrossing populations, three populations with high selfing rates are characterised by a flower morphology that is closer to the selfing syndrome. The presence of outcrossing and selfing floral syndromes within a single species will facilitate unravelling the genetic basis of the selfing syndrome, and addressing which selective forces drive its evolution. / This work was supported by the University of Konstanz (Excellence Initiative Independent Research Startup Grant to MS, http://www.exzellenz.uni-konstanz.de/en/); the Swiss National Science Foundation (grant numbers CRSI33_127155 and Sinergia AVE 31003A_140917, http://www.snf.ch/en/Pages/default.aspx); the University of Zurich (University Research Priority Program Evolution in Action, http://www.uzh.ch/research/priorityprograms/university_en.html); and the Human Frontiers Science Program (Young Investigator Award to KKS, http://www.hfsp.org/). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Anadenanthera é um gênero botânico pertencente à família Mimosaceae e endêmico da América Latina e Caribe. Compreende duas espécies arbóreas tropicais: Anadenanthera colubrina (Vell.) Brenan. (angico, angico vermelho, angico branco, curupay) e Anadenanthera peregrina (L.) Speg. (angico, angico preto, angico de casca, angico do cerrado, yopo ou cohoba). As duas espécies são de ocorrência frequente na paisagem da região de Ribeirão Preto, apresentando-se em aglomerados quase monoespecíficos popularmente conhecidos como angicais. Visando contribuir para futuras medidas conservacionistas, o objetivo deste trabalho foi investigar a diversidade genética, o sistema de reprodução, a estrutura genética espacial e o fluxo gênico contemporâneo de A. colubrina e A. peregrina em angicais da Região de Ribeirão Preto SP usando como ferramenta de análise um conjunto de marcadores moleculares microssatélites (SSR). Para isso, foram construídas duas bibliotecas enriquecidas para microssatélites usando a espécie A. colubrina que resultaram em 20 marcadores SSR testados para a espécie e subsequentemente transferidos para A. peregrina. Desses 20 marcadores, 14 foram polimórficos em cada uma das espécies. Através dessa ferramenta molecular, foi possível realizar os estudos de diversidade genética, endogamia e distribuição genética espacial em A. colubrina e A. peregrina na região de Ribeirão Preto, que acusaram de uma maneira geral, muitas semelhanças entre as duas espécies, bem como entre os angicais de uma mesma espécie. A diferença mais marcante encontrada entre elas foi com relação a estrutura genética espacial, pois todos os angicais de A. colubrina apresentaram forte estruturação, enquanto que os de A. peregrina demonstraram ter uma dispersão aleatória dos indivíduos. O sistema reprodutivo e o fluxo de pólen nas duas espécies foi acessado usando sete marcadores moleculares microssatélites. Para essas análises foram genotipados indivíduos juvenis e adultos (totalizando 352 de A. colubrina e 355 de A. peregrina) presentes nos angicais Acol/PB, Aper/SP255 e Aper/Faz. Através das análises constatou-se que ambas as espécies tem sistema de acasalamento misto, embora A. colubrina tenha apresentado uma proporção maior de autofecundação (tm Acol = 0,619; tm Aper= 0,905). Também foram encontrados elevados índices de cruzamento entre parentes (tm-ts Acol = 0,159; tm-ts Aper = 0,216) e parentesco (coancestria), o que resultou num baixo tamanho efetivo populacional para ambas as espécies. As estimativas das taxas de cruzamentos multilocos individuais apresentaram grande variação nas duas espécies, mostrando a flexibilidade do sistema reprodutivo no gênero Anadenanthera. O número efetivo de doadores de pólen foi muito baixo para um mesmo fruto (1,10 em A. colubrina e 1,24 em A. peregrina) e mais alto entre frutos de uma mesma árvore (2,61 em A. colubrina e 3,35 em A. peregrina), usando a estimativa indireta de correlação de paternidade. Análises de paternidade revelaram distâncias de dispersão de pólen em duas escalas para ambas as espécies. Dessa forma, ocorreram muitos cruzamentos locais, entre árvores próximas no mesmo angical, mas também foram encontradas grandes distâncias de dispersão de pólen. A média da distância de dispersão em A. colubrina foi de 299,88 m e de 214,369 m em A. peregrina. Alto fluxo de pólen oriundo de árvores externas aos angicais de ambas as espécies foi detectado, indicando que os grupos não são isolados reprodutivamente. Por outro lado, o fluxo gênico crítico foi também muito elevado nas estimativas, provavelmente devido ao baixo poder de exclusão que os locos apresentaram dentro dos angicias de ambas as espécies. / Anadenanthera is a genus of Mimosaceae that is endemic to Latin America and the West Indies and comprises two tropical tree species: Anadenanthera colubrina (Vell.) Brenan. (popularly known as angico, angico vermelho, angico branco or curupay) and Anadenanthera peregrina (L.) Speg. (angico, angico preto, angico de casca, angico do cerrado, yopo or cohoba). Both species are commonly found in the Ribeirão Preto region, usually as nearly monospecific agglomerates known as angicais. To aid future conservationist measures, this work investigated the genetic diversity, gene flow, spatial genetic structure and contemporary mating system of A. colubrina and A. peregrina in the angicais of Ribeirão Preto Region SP by analyzing a sample of simple sequence repeat markers (SSR). Two microsatellites libraries were created from A. colubrina, providing 20 SSR markers that were tested for that species and later applied to A. peregrina. Fourteen out of the 20 markers were polymorphic between the species, allowing an examination of the genetic diversity, endogamy and spatial genetic structure in A. colubrina and A. peregrina in the Ribeirão Preto region, which revealed several similarities between the two species, as well as among the angicais of a single species. The most remarkable difference between the species was related to the spatial genetic structure, as all angicais of A. colubrina presented strong structuration, whereas those of A. peregrina exhibited an aleatory dispersion of individuals. The mating system and pollen flow in both species were analyzed through seven SSR. Adults and juveniles from the angicais Acol/PB, Aper/SP255 and Aper/Faz were genotyped for those analyses (352 specimens of A. colubrina and 355 of A. peregrina), revealing that both species undergo a mixed mating system, although A. colubrina presented a higher percentage of self-mating (tm Acol = 0.619; tm Aper= 0.905). High indices of mating among relatives (tm-ts Acol = 0.159; tm-ts Aper = 0.216) and coancestry were also found, resulting in a low effective population size for both species. A wide range in the estimate of the mutilocus breeding rate was found for both species, reflecting the plasticity of the mating system in the genus Anadenanthera. The effective number of pollen donors was very low for a single fruit (1.10 in A. colubrina and 1.24 in A. peregrina) and higher between fruits from the same tree (2.61 in A. colubrina and 3.35 in A. peregrina), using an indirect estimate of the paternity correlation. Paternity analyses revealed the distance of pollen dispersion on two different scales: many local outcrossings (between close trees from the same angical) in addition to long-distance pollen dispersion. The average dispersion distance was 299.88 m in A. colubrina and 214.369 m in A. peregrina. A high pollen flux from trees outside the angicais of both species was observed, indicating a lack of reproductive isolation. However, the gene flow was also very high, likely due to the low power of exclusion presented by loci from both species inside the angicais.

Je m’intéresse à l’écologie évolutive et en particulier à l’évolution des systèmes de reproduction, à l’adaptation et aux interactions de ces deux processus. En effet, les modèles théoriques ainsi qu’un certain nombre d’observations en populations naturelles soulignent que le système de reproduction devrait avoir un effet majeur sur les processus évolutifs et démographiques en populations naturelles. Lors de ma thèse, j’aborde ces thèmes sous divers angles à partir d’une Brassicacée tolérante et hyper-accumulatrice aux métaux lourds, Noccaea caerulescens, et à partir de modèles théoriques. Dans un premier temps, je m’intéresse à la variation du système de reproduction en populations naturelles. Comment s’organisent les flux de gènes entre populations ou sous-populations, et entre individus d’une même population ? Je me suis particulièrement intéressée à l’influence de la pollution des sols sur le taux d’autofécondation chez N. caerulescens, aux flux de gènes entre populations vivant dans les mines et populations vivant sur des sols non contaminés, et à la structure à l’intérieur des populations. Afin de mieux comprendre les facteurs influençant le système de reproduction, j’ai ensuite testé l’effet de la densité en plantes sur le taux d’autofécondations en populations naturelles. Dans un second temps, je teste les interactions existant entre adaptation à des milieux potentiellement très toxiques et système de reproduction. Plus spécifiquement, je teste si la dépression de consanguinité dépend du niveau de stress que subissent les populations, en interaction avec l’histoire des pressions de sélection qu’on subies les populations par le passé. Enfin, à l’aide de simulations, j’étudie comment la variation interannuelle du système de reproduction peut influencer la probabilité d’adaptation de populations faisant face à un changement environnemental. / During my PhD, I focused on the evolution of mating system, adaptation and the mutual influences of both processes. I have been studying different aspects of this interaction using a hypertolerant, hyperaccumulator plant species, Noccaea caerulescens. This species grows on former mines and non-contaminated soils in Europe and in particularly in the Cévennes, and it is an excellent model to study the interaction of local adaptation in a heterogeneous environment and mating system. Firstly, I finely characterized N. caerulescens mating system in natural population, and see how metal pollution affects the variation of mating system in the Cévennes region. I showed that contrary to a couple of classical results (Antonovics 1968), in this system, metallicolous populations have lower self-fertilization rates than nonmetallicolous populations (article submitted). I then tested our best potential factor potentially explaining the variation of mating system in natural populations: plant density. In two different measures, with two different methods, density seems to have no or only a weak effect on self-fertilization rates in Noccaea caerulescens (article in prep). In a second project, I test the interaction between inbreeding depression, stress and the history of adaptation to a given environment using Noccaea caerulescens. Inbreeding depression is known to vary with environment and, sometimes, stress. Both experimental data (Long et al 2013) and theoretical models (Ronce et al 2009) stress the importance of the effect of the history of selection and adaptation in populations on the magnitude of inbreeding depression. Since we have populations of Noccaea caerulescens that are adapted to different levels of pollution, since different levels of pollution impose differential stress on the two ecotypes (strong polution is not good for nonmetallicolous plants) and since the species is self-compatible, this seems like an excellent system to test predictions on the interaction of inbreeding depression and mating system.

Although the process by which fertilization occurs in bryozoans is well described, the ability to self-fertilize and the subsequent ecological consequences are poorly understood. Culturing experiments were conducted examining the effects of selfing on offspring survival and reproduction in the simultaneous hermaphrodite Bugula stolonifera collected from Eel Pond, Woods Hole, MA. Results from these experiments document significant decreases in survival and fecundity of selfed offspring, compared to outcrossed controls, suggesting that these animals are not routinely self-fertilizing in Eel Pond. How these arborescent colonies minimize selfing remains unclear, but it is hypothesized that conspecific aggregations could serve to minimize the chances that a colony utilizes its own sperm for fertilization. The genetic composition of these aggregations was investigated using a newly developed microsatellite library. As larvae routinely metamorphose on conspecific colonies, the possibility that larvae select or avoid their maternal colony was also investigated. Analyses of genetic structure document homogeneity throughout these aggregations on extremely small spatial scales, suggesting high amounts of larval dispersal within aggregations. When combined with results from parentage-exclusion and kinship analyses, these results indicate that a colony's nearest neighbors are not composed of siblings, potentially minimizing inbreeding. Molecular analyses were then used to determine if the high larval dispersal within aggregations resulted in high mixing between aggregations. Sites within Eel Pond separated by 100-300 m were routinely sampled from 2009 to 2011, and analyses were conducted to investigate potential inter- and intra-annual genotypic differentiation within and between aggregations. Results document that although low levels of mixing could result in increased homogeneity between some aggregations, barriers to genetic exchange prevent mixing between most sites. Further, inter-annual comparisons within sites document that significant differentiation can occur between reproductive seasons. Hence, any potential homogeneity achieved between sites during one reproductive season will likely be lost by the beginning of the next reproductive season. Additionally, while sampling in Eel Pond in 2010, I document the first occurrence from the western Atlantic Ocean of another aggregating arborescent bryozoan, Tricellaria inopinata. The growth and reproductive biology of these animals was monitored throughout 2011; results suggest that this introduction is likely to persist.

Der tropische Regenwald im südwestlichen Amazonasgebiet ist durch den zunehmenden Bevölkerungsdruck, die wirtschaftliche Ausbeutung, die Abholzung und das Abbrennen großer Flächen für die Viehwirtschaft bedroht. Allgemein zeichnen sich tropische Regenwälder durch eine hohe Diversität von Baumarten und daraus folgenden niedrigen Populationsdichten und entsprechend hohen Distanzen zwischen den Artgenossen aus. Die räumliche Verteilung der Individuen hat grundlegende Konsequenzen auf die Bewegung der Bestäuber und den intraspezifischen Genfluss, was somit die sexuelle Fortpflanzung beeinflusst. Daher ist das Wissen über den Genfluss grundlegend für das Verständnis von Reproduktionserfolgen und dem Management tropischer Baumarten. In dieser Studie untersuchte ich unterschiedliche Aspekte der sexuellen Fortpflanzung von wildem und kultiviertem Kakao (Theobroma cacao) und zwei weiteren Theobroma-Arten in Bolivien. Folgende Hypothesen wurden angenommen: 1) Wilde und kultivierte Pflanzen von T. cacao in Bolivien sind genetisch unterschiedlich; 2) Die Verteilungsdistanz der Pollen wilder Kakaopflanzen ist größer als die der kultivierten Pflanzen; 3) Es existiert ein gewisser Genfluss zwischen den wilden und den kultivierten Pflanzen von T. cacao; 4) Die Ansammlung der Bestäuber unterscheidet sich zwischen den Populationen wilder und kultivierter Pflanzen von T. cacao, sowohl in Bezug auf die taxonomische Zusammensetzung als auch in der Abundanz der Blütenbesucher; 5) Die kleinräumige genetische Struktur der Theobroma-Arten ist aufgrund der Abhängigkeit von Tieren bezüglich Bestäubung und Samenverbreitung gemäßigt. Zu diesem Zweck wurden im Tiefland von Bolivien drei verbreitete Arten der Gattung Theobroma ausgewählt: T. cacao L. (wild und kultiviert), T. speciosum Willd. ex Spreng und T. subincanum Mart. Diese Arten sind stammblütige Bäume des Unterwuchses. Sie haben zahlreiche kleine Blüten und schwere Früchte mit essbaren Samen, welche von Fruchtfleisch umgeben sind. Um den Pollenfluss des Kakaos zu untersuchen, schätzte ich den Grad der Selbstbestäubung und die Distanz der Pollenverbreitung sowie den Genfluss zwischen wildem und kultiviertem Kakao. Ich untersuchte zudem, ob sich die wilden von den kultivierten Kakaobäumen genetisch unterscheiden. Um die Kakaobestäuber zu identifizieren, dokumentierte ich die Zusammensetzung der Blütenbesucher auf wildem und kultiviertem Kakao. Weiterhin ermittelte ich die genetische Diversität und die kleinräumige genetische Struktur (SGS) der drei Theobroma-Arten, um die Distanzen der genetischen Verbreitung aus den Mustern der SGS zu schätzen. Die Ergebnisse zeigen, dass wilder und kultivierter Kakao sich genetisch unterscheiden, woraus geschlussfolgert wird,  dass die „wilden“ Populationen des Kakaos im Tiefland Boliviens tatsächlich wilde Formen oder zumindest sehr alte Sorten darstellen. Wilde Populationen zeigten eine geringere genetische Diversität als kultivierter Kakao, was möglicherweise durch die unterschiedliche Herkunft der kultivierten Sorten (z.B. Costa Rica, Trinidad y Tobago) begründet ist. Im Unterschied zu früheren Studien konnte ich Hinweise für die Selbstbestäubung in wildem und kultiviertem Kakao finden, auch wenn die Fremdbestäubung klar dominiert. Selbstbestäubung war häufiger in kultiviertem als im wilden Kakao festzustellen. Die gesamtdurchschnittliche Distanz der Pollenverbreitung betrug 867 m. Die Distanz der Pollenverbreitung des wilden Kakaos lag in einem Bereich von 10 und 3007 m, während die der Pollen des kultivierten Kakaos über Distanzen von 13 bis 2360 m verbreitet wurden. Die Bestäubungsdistanzen des Kakaos waren höher als die der typischen Unterwuchsbaumarten, wenn man die geringen Größen der bestäubenden Mücken berücksichtigt. Der relativ hohe Pollenaustausch von kultiviertem zu wildem Kakao (20%) gefährdet die genetische Identität der wilden Populationen. Weiterhin wurde eine große Auswahl an Blütenbesuchern dokumentiert und erhebliche Unterschiede zwischen den Besuchern des wilden und des kultivierten Kakaos festgestellt. Die Ergebnisse lassen vermuten, dass die Mücken als Haupt- oder gar alleinige Bestäuber in unserem Untersuchungsgebiet nicht in Betracht kommen, da sie zu selten vorkommen. Potenzielle zusätzliche Bestäuber sind die kleine Diptera (z.B. Chloropidae und Phoridae) und Hymenoptera (z.B. Eulophidae und Platygastridae). Die wilden Verwandten des T. cacao, T. speciosum und T. subincanum zeigten eine relativ hohe genetische Diversität. Die Arten von Theobroma hatten eine niedrige, aber signifikant kleinräumige genetische Struktur. T. speciosum zeigte die höchste Distanz der Pollenverbreitung, was auf eine effiziente Samen- und Pollenverbreitung hindeutet, wohingegen T. subincanum die strukturiertere Art mit der geringsten Distanz der Genverbreitung war.  Abschließend sollten bei verbessertem Management der Kakao-Populationen folgende Punkte beachtet werden: Die hohe Distanz der Pollenverbreitung bei Kakao und der relativ hohe Pollenaustausch von kultiviertem zu wildem Kakao zeigt die Notwendigkeit des Schutzes weitläufiger Naturwälder. Um die genetische Identität des wilden Kakaos in Bolivien zu schützen und zusätzlich den genetischen Austausch zwischen den wilden Populationen zu unterstützen, sollte die genetische Variabilität lebensfähiger Populationen zu erhalten werden. Die kleinräumige genetische Struktur in Theobroma macht die Bedeutung des Schutzes der samenverbreitenden Tiere deutlich, um die genetischen Ressourcen dieser tropischen Baumgattung zu erhalten.

<p>The evolution of selfing from outcrossing is one of the most frequent mating system transitions in angiosperms. Plants that are highly selfing typically exhibit a suite of morphological traits termed a "selfing syndrome," including reduced corollas and reproductive structures, loss of corolla pigmentation, little anther-stigma separation, and a low pollen/ovule ratio. The overall consensus among scientist is that the morphological changes that accompany the transition to selfing are adaptive and thus a product of natural selection. Few attempts, however, have been made to determine whether traits of the selfing syndrome are truly an operation of natural selection or if genetic drift could be the acting force. My dissertation examines the roles that natural selection and genetic drift played in the evolution of the selfing syndrome in Ipomoea lacunosa. With the use of field observations, crossing data, and molecular analyses, I show that I. lacunosa has evolved increased selfing ability, decreased anther-stigma distance and smaller, white flowers, compared to its closest relative I. cordatotriloba. Furthermore, using a standard QST - FST comparison, I evaluated the relative importance of selection and drift in the evolution of the selfing syndrome in I. lacunosa. I also identified the genetic basis of flower color divergence between I. lacunosa (white) and I. cordatotriloba (purple) and examined patterns of variation to determine if selection or genetic drift caused the divergence. Analyses revealed that the traits of I. lacunosa characteristic of the selfing syndrome have evolved as a product of natural selection, not genetic drift.</p> / Dissertation

Anadenanthera é um gênero botânico pertencente à família Mimosaceae e endêmico da América Latina e Caribe. Compreende duas espécies arbóreas tropicais: Anadenanthera colubrina (Vell.) Brenan. (angico, angico vermelho, angico branco, curupay) e Anadenanthera peregrina (L.) Speg. (angico, angico preto, angico de casca, angico do cerrado, yopo ou cohoba). As duas espécies são de ocorrência frequente na paisagem da região de Ribeirão Preto, apresentando-se em aglomerados quase monoespecíficos popularmente conhecidos como angicais. Visando contribuir para futuras medidas conservacionistas, o objetivo deste trabalho foi investigar a diversidade genética, o sistema de reprodução, a estrutura genética espacial e o fluxo gênico contemporâneo de A. colubrina e A. peregrina em angicais da Região de Ribeirão Preto SP usando como ferramenta de análise um conjunto de marcadores moleculares microssatélites (SSR). Para isso, foram construídas duas bibliotecas enriquecidas para microssatélites usando a espécie A. colubrina que resultaram em 20 marcadores SSR testados para a espécie e subsequentemente transferidos para A. peregrina. Desses 20 marcadores, 14 foram polimórficos em cada uma das espécies. Através dessa ferramenta molecular, foi possível realizar os estudos de diversidade genética, endogamia e distribuição genética espacial em A. colubrina e A. peregrina na região de Ribeirão Preto, que acusaram de uma maneira geral, muitas semelhanças entre as duas espécies, bem como entre os angicais de uma mesma espécie. A diferença mais marcante encontrada entre elas foi com relação a estrutura genética espacial, pois todos os angicais de A. colubrina apresentaram forte estruturação, enquanto que os de A. peregrina demonstraram ter uma dispersão aleatória dos indivíduos. O sistema reprodutivo e o fluxo de pólen nas duas espécies foi acessado usando sete marcadores moleculares microssatélites. Para essas análises foram genotipados indivíduos juvenis e adultos (totalizando 352 de A. colubrina e 355 de A. peregrina) presentes nos angicais Acol/PB, Aper/SP255 e Aper/Faz. Através das análises constatou-se que ambas as espécies tem sistema de acasalamento misto, embora A. colubrina tenha apresentado uma proporção maior de autofecundação (tm Acol = 0,619; tm Aper= 0,905). Também foram encontrados elevados índices de cruzamento entre parentes (tm-ts Acol = 0,159; tm-ts Aper = 0,216) e parentesco (coancestria), o que resultou num baixo tamanho efetivo populacional para ambas as espécies. As estimativas das taxas de cruzamentos multilocos individuais apresentaram grande variação nas duas espécies, mostrando a flexibilidade do sistema reprodutivo no gênero Anadenanthera. O número efetivo de doadores de pólen foi muito baixo para um mesmo fruto (1,10 em A. colubrina e 1,24 em A. peregrina) e mais alto entre frutos de uma mesma árvore (2,61 em A. colubrina e 3,35 em A. peregrina), usando a estimativa indireta de correlação de paternidade. Análises de paternidade revelaram distâncias de dispersão de pólen em duas escalas para ambas as espécies. Dessa forma, ocorreram muitos cruzamentos locais, entre árvores próximas no mesmo angical, mas também foram encontradas grandes distâncias de dispersão de pólen. A média da distância de dispersão em A. colubrina foi de 299,88 m e de 214,369 m em A. peregrina. Alto fluxo de pólen oriundo de árvores externas aos angicais de ambas as espécies foi detectado, indicando que os grupos não são isolados reprodutivamente. Por outro lado, o fluxo gênico crítico foi também muito elevado nas estimativas, provavelmente devido ao baixo poder de exclusão que os locos apresentaram dentro dos angicias de ambas as espécies. / Anadenanthera is a genus of Mimosaceae that is endemic to Latin America and the West Indies and comprises two tropical tree species: Anadenanthera colubrina (Vell.) Brenan. (popularly known as angico, angico vermelho, angico branco or curupay) and Anadenanthera peregrina (L.) Speg. (angico, angico preto, angico de casca, angico do cerrado, yopo or cohoba). Both species are commonly found in the Ribeirão Preto region, usually as nearly monospecific agglomerates known as angicais. To aid future conservationist measures, this work investigated the genetic diversity, gene flow, spatial genetic structure and contemporary mating system of A. colubrina and A. peregrina in the angicais of Ribeirão Preto Region SP by analyzing a sample of simple sequence repeat markers (SSR). Two microsatellites libraries were created from A. colubrina, providing 20 SSR markers that were tested for that species and later applied to A. peregrina. Fourteen out of the 20 markers were polymorphic between the species, allowing an examination of the genetic diversity, endogamy and spatial genetic structure in A. colubrina and A. peregrina in the Ribeirão Preto region, which revealed several similarities between the two species, as well as among the angicais of a single species. The most remarkable difference between the species was related to the spatial genetic structure, as all angicais of A. colubrina presented strong structuration, whereas those of A. peregrina exhibited an aleatory dispersion of individuals. The mating system and pollen flow in both species were analyzed through seven SSR. Adults and juveniles from the angicais Acol/PB, Aper/SP255 and Aper/Faz were genotyped for those analyses (352 specimens of A. colubrina and 355 of A. peregrina), revealing that both species undergo a mixed mating system, although A. colubrina presented a higher percentage of self-mating (tm Acol = 0.619; tm Aper= 0.905). High indices of mating among relatives (tm-ts Acol = 0.159; tm-ts Aper = 0.216) and coancestry were also found, resulting in a low effective population size for both species. A wide range in the estimate of the mutilocus breeding rate was found for both species, reflecting the plasticity of the mating system in the genus Anadenanthera. The effective number of pollen donors was very low for a single fruit (1.10 in A. colubrina and 1.24 in A. peregrina) and higher between fruits from the same tree (2.61 in A. colubrina and 3.35 in A. peregrina), using an indirect estimate of the paternity correlation. Paternity analyses revealed the distance of pollen dispersion on two different scales: many local outcrossings (between close trees from the same angical) in addition to long-distance pollen dispersion. The average dispersion distance was 299.88 m in A. colubrina and 214.369 m in A. peregrina. A high pollen flux from trees outside the angicais of both species was observed, indicating a lack of reproductive isolation. However, the gene flow was also very high, likely due to the low power of exclusion presented by loci from both species inside the angicais.

Habitat fragmentation is one of the main threats to semi-natural grasslands all over Europe, causing reductions in both numbers of populations and numbers of individuals withinpopulations. Small isolated populations are particularly vulnerable to fluctuations that reducespopulation size and could lead to loss of genetic variation due to inbreeding depression, orfixation of harmful alleles due to genetic drift. The aim of this study was to investigate if thecritically endangered eyebright Euphrasia stricta var. suecica experiences pollen limitation,inbreeding depression and heterosis or outbreeding depression. Low pollen limitation waspredicted as well as low inbreeding depression due to high selfing rate. Outbreedingdepression or heterosis was expected based on previous studies indicating strong geneticdifferentiation among populations. This study was conducted in two traditionally managedmeadows in the central part of Gotland, Sweden, using supplemental hand-pollinations andcontrolled crosses. Individuals were collected when the flowering period was over and seedswere counted and weighed. There were no signs of either pollen limitation, inbreedingdepression, heterosis or outbreeding depression in any of the two wooded hay meadows. Thedegree of autonomous selfing was high, indicating that E. stricta var. suecica is not dependenton pollinators for seed production. A significant difference between the meadows was foundin proportion of viable seeds. As there were no signs of local adaptation, individuals could betranslocated between the two studied populations or seed sowings could be used to increasepopulation size. However, before such conservation efforts are undertaken, it is important toconduct further studies that investigate several populations including more life-stages.