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Diplomarbeit DIPLOMARBEIT Titel der Diplomarbeit Populationsgenetik des Chloroplastengenoms agamospermer und sexueller Potentilla-Arten (Rosaceae) angestrebter akademischer Grad Magistra der Pharmazie (Mag.pharm.) Verfasserin: Hagenhofer Judith Matrikel-Nummer: a0306986 Studienrichtung A449 Pharmazie (lt. Studien-blatt): Betreuer: Ao. Univ. Prof. Dr. Johannes Saukel Wien, im August 2011 Danksagung An dieser Stelle möchte ich mich herzlichst bei meinem Diplomarbeitsbetreuer Dr. Christoph Dobeš bedanken. Ohne seine Hilfe, Beratung, und sein fundiertes Wissen, wäre die Diplomarbeit in der Form nicht möglich gewesen. Mein Dank gilt auch Herrn ao. Univ.-Prof. Dr. Johannes Saukel für die Bereitstellung des Themas und für seine Unterstützung in statistischen Fragen. Auch möchte ich an dieser Stelle Dipl. Biol. Valerie Klatte-Asselmeyer und Mag. Susanne Scheffknecht danken, die mir in vielen Angelegenheiten beistanden und die zu einem sehr großen Teil für das angenehme Arbeitsklima verantwortlich waren. Ein großer Dank gilt Dr. Mag. Fatemeh Maghuly für die vielen Antworten auf meine phylogenetischen Fragen. Ein weiterer großer Dank gilt ao. Univ. Prof. Dr. phil. Margit Laimer Da Camara Machado. Danke Margit für alles. Einen ganz ganz großen Dank möchte ich auch Betti und Po aussprechen, die mir beide beim Erstellen meiner Graphiken sehr geholfen haben. Auch möchte ich Dr. Huber hier erwähnen für das Herstellen einer Internetverbindung, wenn dies mir nicht möglich war, aber trotzdem dringend gebraucht wurde. Dankeschön auch euch ,Stefan und Leni, fürs Anteilnehmen an meiner Diplomarbeit. Ein besonderer Dank gilt meiner Mutter, ohne die das Studium nicht möglich gewesen wäre. Und last but not least danke ich Caterina für die grenzenlose moralische Unterstützung. Inhaltsverzeichnis 1 Einleitung ............................................................................................................. 1 1.1 Die Gattung Potentilla ......................................................................................... 1 1.1.1 Taxonomie der Gattung Potentilla nach morphologischer Betrachtung (Wolf 1908) .......................................................................................................... 2 1.1.2 Taxonomie der Gattung Potentilla nach phylogenetischer Betrachtungsweise ............................................................................................ 10 1.2 Reproduktionsmechanismus ............................................................................. 12 1.2.1 Sexuelle Fortpflanzung ...................................................................................... 12 1.2.2 Asexuelle Fortpflanzung .................................................................................... 14 1.2.3 Hybridisierung ................................................................................................... 17 1.3 Muster und genetische Variation ...................................................................... 20 1.3.1 Entstehung genetischer Muster und deren Verteilung .................................... 20 1.3.2 Biogeographie ................................................................................................... 21 1.3.3 Phylogeographie ................................................................................................ 22 2 Zielsetzung der Diplomarbeit ............................................................................ 24 3 Material und Methodik ..................................................................................... 25 3.1 Pflanzenmaterial ................................................................................................ 25 3.2 Methoden .......................................................................................................... 27 3.2.1 DNA Extraktion der zu untersuchenden Pflanzen ............................................. 27 3.2.2 PCR ..................................................................................................................... 29 3.2.3 Sequenzierung trnC-ycf6 IGS ............................................................................. 30 3.2.4 Assembling und Alignment ............................................................................... 32 3.2.5 Phylogenetische Analyse ................................................................................... 33 3.2.6 Diversität ........................................................................................................... 34 4 Ergebnisse .......................................................................................................... 35 4.1 Vergleich unterschiedlicher Polymerasen ........................................................ 35 4.2 Sequenzierung des trnC-ycf6 IGS ....................................................................... 35 4.3 Sequenzvariabilität ............................................................................................. 36 4.4 Bestimmung der Haplotypen ............................................................................. 40 4.4.1 Längenvariationen der Haplotypen: .................................................................. 41 4.5 Plastid sharing .................................................................................................... 42 4.6 Phylogenetische Beziehungen ........................................................................... 47 4.6.1 Maximum-Parsimony Stammbaum ................................................................... 48 4.6.2 Maximum-Parsimony Stammbaum ................................................................... 50 4.6.2 Majority 50%- Stammbaum ............................................................................... 52 4.7 Gen- und Nukleotid-Diversität .......................................................................... 54 4.7.1 Haplotypendiversität der Arten und Populationen ........................................... 54 4.7.2 Nukleotid- Diversität .......................................................................................... 55 4.8 Geographische Verbreitung der Diversitäten ................................................... 56 4.8.1 Geographie der Gen-Diversitäten ...................................................................... 56 4.8.2 Geographische Verbreitung der Haplotypen..................................................... 60 5 Diskussion .......................................................................................................... 64 5.1 Polymerase–Effizienz ......................................................................................... 64 5.2 Plastid-sharing .................................................................................................... 65 5.3 Phylogenie .......................................................................................................... 66 5.4 Genetische Diversität ......................................................................................... 67 6 Zusammenfassung ............................................................................................. 69 7 Summary ............................................................................................................ 71 8 Tabellenverzeichnis ........................................................................................... 72 9 Abbildungsverzeichnis ....................................................................................... 73 10 Quellenverzeichnis ............................................................................................ 74 11 Anhang............................................................................................................... 81 11.1 Chemikalien und Reagenzien ............................................................................ 81 11.2 Elektrophorese .................................................................................................. 81 11.3 PCR ..................................................................................................................... 82 11.3.1 Mastermixzusammensetzung ........................................................................... 82 11.3.2 PCR-Programm .................................................................................................. 83 11.4 Sequenzierung .................................................................................................. 83 11.4.1 Mastermixzusammensetzung ........................................................................... 83 11.4.2 Programm.......................................................................................................... 84 11.5 Populationsdiversitäten trnC-ycf6 cp-DNA Haplotypen ................................... 85 11.6 Populationsdiversiäten der Haplotypen basiered auf dem .................................. Gesamtalignment .............................................................................................. 93 11.7 Mittelwerte der Gen- bzw Nukleotid- Diversität .............................................. 97 11.8 Liste der Haplotypen, deren Vorkommen sowie Häufigkeiten ............................ des Auftretens ................................................................................................... 99 11.9 Materialbestandsliste (DFG DO 792/1-1 von Dr. Christoph Dobeš) ............... 111 11.10 Materialbestandsliste ...................................................................................... 112 12 Lebenslauf ......................................................................................................
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