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Microtus Agrestis): INSIGHTS from GENETIC, KARYOTYPIC and MORPHOMETRIC DATA

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Microtus Agrestis): INSIGHTS from GENETIC, KARYOTYPIC and MORPHOMETRIC DATA CRYPTIC SPECIATION IN THE FIELD VOLE (Microtus agrestis): INSIGHTS FROM GENETIC, KARYOTYPIC AND MORPHOMETRIC DATA Joana Maria Jorge Pereira de Castro Paupério Doutoramento em Biologia Departamento de Biologia 2012 Orientador Paulo Célio Alves, Professor Auxiliar, Faculdade de Ciências da Universidade do Porto Coorientador Jeremy Byron Searle, Professor, University of York Ao Adriano e às minhas três princesas, Nota prévia: Na elaboração desta dissertação, e nos termos do número 2 do Artigo 4º do Regulamento Geral dos Terceiros Ciclos de Estudos da Universidade do Porto e do Artigo 31º do D.L. 74/2006, de 24 de Março, com a nova redação introduzida pelo D.L. 230/2009, de 14 de Setembro, foi efetuado o aproveitamento total de um conjunto coerente de trabalhos de investigação já publicados ou submetidos para publicação em revistas internacionais indexadas e com arbitragem científica, os quais integram alguns dos capítulos da presente tese. Tendo em conta que os referidos trabalhos foram realizados com a colaboração de outros autores, o candidato esclarece que, em todos eles, participou ativamente na sua conceção, na obtenção, análise e discussão de resultados, bem como na elaboração da sua forma publicada. This research was supported by Programa Operacional Potencial Humano – Quadro de Referência Estratégico National funds from the European Social fund and the Portuguese Ministério de Educação e Ciência through a PhD fellowship (Fundação para a Ciência e a Tecnologia, FCT - SFRH/BD/27369/2006). This work was developed in This thesis should be cited as: Paupério J (2012) Cryptic speciation in the field vole (Microtus agrestis): insights from genetic, karyotypic and morphometric data. PhD Thesis, University of Porto, Porto, Portugal. ACKNOWLEDGEMENTS Lots of people contributed to the content of this thesis. Either by giving samples, suggestions or support, or by making me think about things or just by being present... To all I would like to say: Thank you! I would like to thank my two supervisors, Paulo and Jeremy, without them this thesis would not have been accomplished. Paulo, já me aturas há bastante tempo… há mais de uma década!!! E, na verdade és totalmente responsável pelo tema desta tese. Foste tu que me puseste o ’bichinho do agrestis atrás da orelha’... mostraste-me os primeiros trabalhos realizados pela Maarit com esta espécie… Obrigada por isso! Obrigada também por me teres acompanhado todos estes anos, por todas as tuas sugestões, ensinamentos, pelo teu companheirismo, apoio e amizade e também pelo entusiasmo que sempre demonstraste com este trabalho e que sempre me deu grande ânimo. Jeremy, it has been a pleasure to know you and work with you! Thanks a lot for being so welcoming and for sharing with me all your knowledge and experience not only in the lab but also in our field trips. You were my good luck charm for finding voles…The voles seemed to throw themselves into the pool at CIBIO whenever you were in town! I enjoyed a lot my time in York, specially our nice suppers with Sofia’s chocolate mousse and our memorable conversations about pudding... ups, sorry, I meant to say the continuous hard work! Thank you for being such a good supervisor! I would also like to thank the institutions that made this work possible. To FCT, for the attribution of my PhD fellowship. To CIBIO and Professor Nuno Ferrand for the opportunity of developing this study in this excellent research centre. And to the University of York, for welcoming me during my stays. I also want to express my appreciation to the institutions that gave me the appropriate permits for trapping and collecting tissue samples from field voles both in Portugal and northern Spain: Instituto da Conservação da Natureza e da Biodiversidade, Consejería de Medio Ambiente do Gobierno del Principado de Asturias, Consellería de Medio Ambiente y Desenvolvemento Sostible de Galicia, Servició Territorial de Medio Ambiente de la Comunidad Autónoma de Castilla Léon, Parque Nacional Picos da Europa, Dirección General de Biodiversidad de la Comunidad Autónoma de Cantabria, Departamento de Agricultura de la Deputación Foral de Bizkaia, Departamento de Medio Ambiente de la Diputación Foral de Álava, Dirección General de Medio Natural da Comunidad Autónoma de Cataluña. The results presented in this thesis wouldn’t have been accomplished without the contribution of a great number of people and institutions. First of all to Maarit Jaarola, thank you for your availability in answering my doubts and sending me some of your samples. It was a pleasure to have you as a co-author of the first paper of the thesis! To Alejandro Centeno-Cuadros, Alfonso Ceña, Antoni Arrizabalaga and the Museum of Natural Science of Granollers, António Mira and the PERSIST project team, Augusto de Castro, Beatriz e Vasco Alves, Carlos Fonseca, Carlos Nores, Cristianne Bastos-Silveira, Enrique Castiéns, Fernando Jubete, Guilherme Dias, Hélia Gonçalves, Jacinto Róman, José António Godoy, José Manuel Grosso-Silva, Maria José Lopez-Fuster and the University of Barcelona, the Portuguese National Tissue Bank/National Institute for Nature and Biodiversity Conservation (BTVS/ICNB), Rocio Rosa-García, Sara Afonso, Vanessa Mata, Verónica Gomes and Zbyszek Boratynski, thanks a lot for providing samples for this study! I am also grateful to Maria da Luz Mathias and the University of Lisbon, for lending some field materials. And, as not only people may contribute to a PhD thesis, I would also like to express my gratitude in a special way to Rafi (Paulo’s dog), Tomás and Manuchau (Pedro’s cats) and Fran (Guilherme’s cat). To the York gang, Mabel, Sofia, Ellie, Rodrigo, Sophie, Natalia, Jerry, Dan, Frida, Bruce, Mike, Nancy and Ollie thank you guys for being so welcoming! Despite the rainy weather, I really enjoyed the time I spend in York! Jerry, thank you for teaching me the ways around the lab and for getting me started in the molecular analysis. I was very happy that we could collaborate in part of the work developed in the thesis and hopefully we will continue to work together in the next projects. Natalia, the other mum in the lab, with you I shared my family homesickness and I learned a lot, especially how to be efficient in the lab! Thank you for that! Rodrigo, I really enjoyed our field trip in northern Spain. We’ve seen very nice places and trapped lots of voles, and some shrews… Thank you for all our conservations and discussions, I learned a lot with that. Mabelita, cariño… how nice was your company… I couldn’t have done it without you! Thank you for all the moments we spent at the lab and in the city… thanks a lot for keeping me company. Thanks for teaching me all about the karyotypes and working with me on the paper, it turned out to be a very promising work! I really enjoyed your visit to Porto and hope you can come and visit us soon! Sofia, a minha companheira portuguesa nas terras britânicas! Que boas as nossas conversas, os nossos jantares e passeios na cidade… Obrigado por me ajudares a superar as saudades de casa e da minha família! Com a tua companhia tudo foi mais fácil! Obrigado por tudo!!!! Aos meus colegas do CIBIO, em especial ao gabinete do GEPE mas não só, um muito obrigado. Sónia, Peter spider man, Xico, Francisco, Raquel, Cândida, Ana, Fátima, João, Verónica, Tiago meus colegas de gabinete ou no lab, em um ou outro momento muito obrigado pelas conversas, dicas e pela companhia. Tarroso, obrigada pelas eternas ajudas no arcgis! À Ana Silva, Alexandra, Joana, Rómulo, agradeço a ajuda no trabalho de campo e na análise das egagrópilas. Ao Zef e ao Miguel Carneiro, muito obrigado pelas conversas e discussões sobre species trees e evolução dos Microtus. Zef, a tua participação nessa fase do trabalho foi essencial. À Sandra e à Sara muito obrigado por estarem sempre disponíveis e fazerem sempre todos os possíveis para tornarem o nosso trabalho mais fácil! Aos colegas do lab da FCUP (Tiago, Sandra, Pedro e Zé Manel) obrigadinha por me terem acompanhado nestes últimos momentos de escrita. Tiago, obrigada pelo apoio, por me responderes às minhas dúvidas, mesmo as mais idiotas, pelos teus treinos de relaxamento… Às meninas do CTM (e meninos) muito obrigado por me terem acolhido quando precisei de ir analisar os meus crânios. Diana, obrigado por tudo o que me ensinaste, a tua experiência foi fundamental para o sucesso deste trabalho! À Hélia, por toda a ajuda que me deste no campo, pela amizade e por todas as amostras que me conseguiste, um muito obrigado. És a minha colaboradora nº 1. Ao Ricardo Pita e ao António Mira (Universidade de Évora) por todo o apoio que sempre me deram. António, obrigada pela disponibilidade e pela maneira como sempre me apoiaste neste trabalho. Ricardo, obrigada por me ensinares a capturar os Microtus…as dicas que me deste foram essenciais para o sucesso das minhas saídas de campo. À Soraia, tem sido um prazer trabalhar contigo! O teu companheirismo, amizade e ajuda foram fundamentais para este trabalho! Obrigado! Catas, companheira! Demos os nossos primeiros passos na ciência juntas e apesar de termos seguido caminhos um pouco diferentes estiveste sempre presente. Muito obrigado pela tua amizade, pelos momentos de hiperventilação conjunta e por estares sempre disponível mesmo nos momentos menos bons. A todos os meus amigos, que não vou nomear (eles sabem quem são) muito obrigado por serem quem são e por estarem presentes! Obrigado, Mariana Sampaio, pelo desenho maravilhoso que fizeste para a minha capa. E finalmente, aos meus pais, por todo o apoio que sempre me deram! Sem eles não estaria aqui hoje. A toda a minha família pela amizade e ajuda que sempre nos deram.
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