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Sin Título De Diapositiva SISTEMÀTICA MOLECULAR, FILOGEOGRAFIA I GENÈTICA DE LA CONSERVACIÓ DE MUSTÈLIDS I DE MACACS Josep Marmi i Plana Tesi Doctoral 2004 Departament de Ciències Experimentals i de la Salut UNIVERSITAT POMPEU FABRA Sistemàtica molecular, filogeografia i genètica de la conservació de mustèlids i de macacs Memòria presentada per Josep Mª Marmi Plana per optar al grau de Doctor per la Universitat Pompeu Fabra. Aquest treball ha estat realitzat sota la direcció del Dr. Xavier Domingo i Roura al Departament de Ciències Experimentals i de la Salut de la Universitat Pompeu Fabra. Dr. Xavier Domingo i Roura Josep Mª Marmi i Plana Director de Tesi Barcelona, març de 2004 Dipòsit legal: B.43715-2004 ISBN: 84-688-8523-1 Als meus pares per haver-me ajudat tant durant tots aquests anys Als meus germans A la Queralt A GRAÏMENTS En aquest apartat vull agrair l’ajuda que m’han ofert tot un seguit de persones des que em vaig començar a introduir en el món de la genètica de poblacions i de l’evolució molecular. Sobretot vull agrair als Drs Alfredo Ruíz i Xavier Domingo que em donessin una oportunitat per iniciar-me productivament en aquestes especialitats. Sota la supervisió del primer vaig cursar - juntament amb el col·lega Oscar Ramírez i dins de la llicenciatura de Biologia - l’assignatura de Pràctiques en Empreses i Institucions Públiques amb l’objectiu de realitzar un treball recopilatori d’estudis de diversitat genètica en fauna dels Països Catalans. Amb el segon, després de col·laborar- hi durant una mica més d’un any (entre el setembre de 1998 i el desembre de 1999), vaig iniciar la tesi doctoral que ara presento. Al setembre de 1998 vaig començar a treballar amb en Xavier, en el Centre de Recerca Ecològica i Aplicacions Forestals (CREAF) de la Universitat Autònoma de Barcelona (UAB). En aquesta institució, en Guerau Fernández i la Begonya Jaén van ser els primers que em van ajudar a aprendre les tècniques de biologia molecular que havia d’utilitzar per fer els experiments i en teoria estava supervisat per la Rogelia Lorente, una estudiant de màster mexicana molt trempada però que sempre desapareixia quan més la necessitava (Murphy??), doncs aquell any en Xavier començava a treballar a la Universitat Pompeu Fabra . Al febrer de 1999 en Xavier em va proposar incorporar-me a la Unitat de Biologia Evolutiva de la Universitat Pompeu Fabra. Els companys de la Unitat de Biologia Evolutiva m’han ajudat a millorar les tècniques apreses i me n’han ensenyat de noves amb molta paciència, dedicació i amabilitat. Ells són l’Aida Andrés, la Stephanie Plaza, en David Comas, l’Elena Bosch, l’Eva Mateu. Dins d’aquesta unitat vull agrair especialment l’ajuda oferida per la Mònica Vallès en els treballs d’extracció d’ADN de les mostres; pel Jordi Clarimón que m’ha explicat tots els secrets de les electroforesis en gels de poliacrilamida; pel Bernal Morera per haver pescat algunes de les seqüències de GenBank que he utilitzat; per l’Anna Pérez-Lezaun, per les vegades que m’ha ajudat a seqüenciar (també per les vegades que m’ha donat suport moral després dels terribles atacs d’ira de la Rosa Martínez) juntament amb en Roger Anglada (el deixeble de l’Anna en el Servei de Seqüenciació i d’Anàlisi de Fragments). Sóc conscient que dec uns quants pernils “jabugos” als cervells de la casa (que fan la seva vida a la “sala de pensar”). En Francesc Calafell, l’Arcadi Navarro i l’Oscar Lao han tingut una paciència digna d’admirar amb mi i m’han ofert una ajuda impagable en les anàlisis de dades. També vull agrair en aquestes línies l’amistat que m’han ofert la Marta Soldevila, que ja es va convertir en una molt bona amiga meva quan estudiàvem a la UAB, i en Carles Lalueza-Fox, del qual admiro la seva saviesa i els seus treballs. En aquest darrer any s’han incorporat noves persones a la Unitat, la Lourdes Sampietro, l’Anna Ramírez, en Tomàs Marquès, l’Anna González, la Michelle Gardner, l’Andrés Moreno, la Gemma Berniell, l’Anna Ferrer, la Cristina Santa i en Ferran Casals, entre d’altres, que també s’han convertit en uns bons companys. Les següents línies van dedicades a aquells que es van afegir posteriorment al projecte d’en Xavier després d’en Guerau, la Begonya, la Rogelia, la Rachel Whiteley i jo mateix. Començant una altra vegada pel Guerau (que es va reincorporar temporalment però ara a la UPF) i l’Anna Belém Ibarz amb els que vaig coincidir durant els primers anys a la UPF i que ara han seguit camins diferents. L’Anna actualment és a Oxford amb la intenció de començar un doctorat i en Guerau ja el té encarrilat a l’Hospital de Can Ruti. A ells dos els desitjo un munt d’èxits al llarg dels seus respectius camins. El següent en incorporar-se va ser en Joan Francesc López, un dels grans amics que he fet en aquesta casa i una de les persones que més m’ha ajudat. L’altra és l’Aïnhoa Ferrando a la qual també li dec molt, sobretot després d’ajudar-me amb les delicades extraccions dels pèls de les brotxes d’afaitar. He de reconèixer que després de la incorporació d’ells dos la qualitat del treball del grup va millorar notablement. L’última onada de fitxatges va arribar amb l’aprovació del Projecte Europeu ”INPRIMAT”. Arrel d’això es van incorporar en el nostre grupet, a partir del novembre de 2002, l’Olga Andrés i en Francesc Bisbal. Tots ells també han demostrat ser uns bons companys de treball i el que és més important, uns bons amics. Cal dir aquí que l’Olga ha fet la feixuga feina de llegir-se aquesta tesi per corregir les faltes d’ortografia i gramàtica que a vegades se m’escapen. Al mateix temps que l’Olga, ens va arribar l’ajuda celestial del Bruno Guallar (el tito), com a gestor de projectes. A part d’encarregar-se de resoldre els “marrons de grup” que suposen la paperassa i la sol·licitud de projectes, m’ha ofert una valuosa ajuda en moltes ocasions. És com la nostra padrina i ell m’ha ajudat, per exemple, a fer notables progressos en el disseny i composició de seminaris amb el “Power Point”. Almenys ara les meves presentacions no són tan horteres com ho eren al principi. En Bruno també ha tingut la santa paciència d’ajudar-me en els múltiples i variats problemes de format que he patit a l’etapa final de la tesi que teniu a les vostres mans. El setembre del 2003 s’ha incorporat al nostre grup en Thomas Kellermann, un estudiant alemany molt trempat, i més recentment, el mes de novembre, la Montserrat Bosch ha començat a treballar amb nosaltres com a “postdoc”. Ella m’ha donat consells molt útils per acabar el meu recorregut pre-doctoral. També agraeixo a l’Amarant Martínez, estudiant de la UPF a punt de llicenciar, l’ajuda oferta en el treball de la filogeografia del teixó i a un dels grans amics que he fet a fora del nostre grup, en Luís Magano li agraeixo el seu companyerisme i li desitjo molta sort en la seva nova feina. Per acabar, també tinc en compte l’ajuda desinteressada que m’ha ofert l’amic i biòleg Pere Aymerich, el qual, a part de deixar- me alguns llibres ben interessants, m’ha ensenyat, en diverses tertúlies de cafè, alguns punts de vista sobre el treball de camp que sovint els moleculars desconeixem. Vull dir també, que la realització d’aquesta tesi no hagués estat possible sense el finançament de les següents institucions - Ministerio de Educación y Cultura (Ref. PB98-1064); Institut d’Estudis Catalans; European Commission (Ref. INPRIMAT QLRI-CT-2002-01325); Departament d’Universitats, Recerca i Societat de la Informació, Generalitat de Catalunya (Ref. 2001SGR 00285); People’s Trust for Endangered Species, United Kingdom; Parfumerie Douglas Netherland B.V.; Fundació Territori i Paisatge, Caixa de Catalunya - la beca pre-doctoral que he gaudit durant aquests quatre anys concedida pel Departament d’Universitats, Recerca i Societat de la Informació, Generalitat de Catalunya (Ref. 2000FI-00698) – i tot un seguit de col·laboradors que ens han proporcionat mostres i que estan citats en els diferents capítols. Per acabar, també vull donar les gràcies a l’Eulàlia Garcia-Franquesa, conservadora de vertebrats del Museu de Zoologia de Barcelona, per haver acudit als assatjos de la defensa pública d’aquesta tesi i ajudar-me a millorar la meva presentació. ÍN DEX PRESENTACIÓ 11 INTRODUCCIÓ 15 1. La classificació dels éssers vivents i la conservació de la biodiversitat 1.1 Cap a on va la taxonomia actual? 16 1.2 Caràcters morfològics i caràcters moleculars 19 1.3 El paper de la variació intraespecífica 23 1.4 La sistemàtica molecular i la conservació de la fauna 27 2. Els marcadors genètics 2.1 Genoma mitocondrial i genoma nuclear 33 2.2 El gen citocrom b 35 2.3 Les regions flanquejants de regions repetitives nuclears 36 2.4 La regió control de l’ADN mitocondrial 38 3. Els mustèlids 3.1 Distribució, descripció i història evolutiva 42 3.2 Taxonomia confusa 47 3.3 Estatus i conservació a nivell mundial 49 4. El teixó euroasiàtic 4.1 Distribució, descripció i història evolutiva 50 4.2 Importants diferencies morfològiques 51 4.3 Estatus i conservació 53 5. Els macacs 5.1 Distribució, descripció, taxonomia i història evolutiva 54 5.2 Història evolutiva del grup “Fascicularis” 56 5.3 Els genomes mitocondrial i nuclear discrepen sobre l’evolució de les espècies de macacs 58 5.4 Els macacs rhesus i japonès 60 OBJECTIUS 63 RESULTATS 67 Capítol I: Phylogeny, Evolutionary History and Taxonomy of the Mustelidae Based on Sequences of the cyt b Gene and a Complex Repetitive Flanking Region 69 Capítol II: Badgers Are Divided into Four Clearly Separated Taxonomic Units Across Eurasia 101 Capítol III: Inquiring into the Origin of Badger Hair Used in the Manufacturing of Shaving Brushes 125 Capítol IV: Radiation and Phylogeography in the Japanese Macaque, Macaca fuscata 137 DISCUSSIÓ GENERAL 149 1.
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