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Molekulargenetische Untersuchungen Zur Variabilität Der Fichte (Picea Abies [L.] Karst.) in Deutschland

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Molekulargenetische Untersuchungen Zur Variabilität Der Fichte (Picea Abies [L.] Karst.) in Deutschland Vom Fachbereich VI (Geographie/Geowissenschaften) der Universität Trier zur Erlangung des akademischen Grades Doktor der Naturwissenschaften (Dr. rer. nat.) genehmigte Dissertation Molekulargenetische Untersuchungen zur Variabilität der Fichte (Picea abies [L.] Karst.) in Deutschland vorgelegt von Markus Quack Trier, 2004 Betreuer: Univ.-Prof. Dr. Dr. h.c. mult. Paul Müller Berichterstattende: Univ.-Prof. Dr. Dr. h.c. mult. Paul Müller, PD Dr. Martin Paulus Datum der Disputation: 28.04.2004 Danksagung Das Arbeiten mit sich ständig weiterentwickelnden modernen Verfahren ist spannend und motivierend, erfordert aber auch viel Ausdauer und den Mut zum richtigen Zeitpunkt einen Schlussstrich zu ziehen. Da mir diesbezüglich Anstöße von Freunden und Kollegen hilfreich waren, möchte ich allen danken, die zum Gelingen dieser Arbeit beigetragen haben. An erster Stelle geht mein Dank an Herrn Prof. Dr. Dr. h.c. mult. Paul Müller, dass er mir diese Arbeit ermöglichte und mir bei der Durchführung jeglichen Freiraum ließ. Den Herren Dr. V. Konnert (Nationalparkverwaltung Berchtesgaden), K. Freyer (Bayerische Staats- forstverwaltung), Dr. H. Rall (Nationalparkverwaltung Bayerischer Wald), A. Rommerskirchen (Nationalparkverwaltung Hochharz) möchte ich für Unterstützung bei den Recherchen zur Herkunft der Fichtenbestände von Berchtesgaden, Bayerischem Wald und Harz danken. Den Herren PD Dr. Martin Paulus und apl. Prof. Dr. Willy Werner danke ich für die Durchführung der Probenahmen in Belau und im Hunsrück. Im Rahmen dessen möchte ich mich auch bei allen Kollegen der Umweltprobenbank des Bundes für die Unterstützung bei den Probenahmen sowie für das gute Arbeitsklima und für den „Spaß an der Arbeit“ bedanken. Herrn Dr. Ralf Kautenburger danke ich für seine ständige Hilfsbereitschaft und für die Beantwortung zahlreicher labortechnischer und chemischer Fragen. Für die kritische Durchsicht des Manuskriptes möchte ich mich ganz besonders bei Herrn PD Dr. Roland Klein bedanken, dessen Diskussionsbereitschaft ich sicherlich hätte häufiger nutzen können. Ganz besonderer Dank gilt meiner Frau Judith und meinen Eltern für die Unterstützung während der vergangenen Jahre. Inhalt I Inhaltsverzeichnis 1 Problemstellung und Zielsetzung ........................................................................1 2 Stand der Forschung ............................................................................................5 2.1 Systematik, Einwanderungsgeschichte und Verbreitung .............................................................5 2.2 Standortfaktoren .........................................................................................................................10 2.3 Morphologische Variabilität der Fichte .......................................................................................11 2.4 Genetische Variabilität der Fichte...............................................................................................15 2.4.1 Isoenzymanalysen................................................................................................................15 2.4.2 DNA-gestützte Verfahren .....................................................................................................17 2.5 Geeignete Markertypen ..............................................................................................................19 2.5.1 Random amplified polymorphic DNA (RAPD) ......................................................................25 2.5.2 Inter simple sequence repeats (ISSR) .................................................................................27 2.5.3 Simple sequence repeats (SSR) ..........................................................................................28 2.5.4 Expressed sequence tags (EST)..........................................................................................30 3 Untersuchungsgebiete und Beschreibung der Untersuchungsflächen.........32 3.1 Untersuchungsgebiete innerhalb des natürlichen Verbreitungsgebietes...................................35 3.1.1 Nationalpark Berchtesgaden ................................................................................................35 3.1.2 Nationalpark Bayerischer Wald ............................................................................................36 3.1.3 Nationalpark Hochharz.........................................................................................................38 3.2 Untersuchungsgebiete außerhalb des natürlichen Verbreitungsgebietes .................................39 3.2.1 Saarländischer Verdichtungsraum (Warndt) ........................................................................39 3.2.2 Hunsrück...............................................................................................................................41 3.2.3 Naturpark Solling-Vogler ......................................................................................................41 3.2.4 Bornhöveder Seengebiet (Belauer See) ..............................................................................42 4 Material und Methoden .......................................................................................44 4.1 Material .......................................................................................................................................44 4.1.1 Probenmaterial .....................................................................................................................44 4.1.2 Puffer und Lösungen ............................................................................................................45 4.2 Methoden....................................................................................................................................45 4.2.1 Morphologische und biometrische Charakterisierung ..........................................................45 4.2.2 DNA-Isolierung .....................................................................................................................46 4.2.3 RAPD-Analyse......................................................................................................................46 4.2.4 ISSR-Analyse .......................................................................................................................49 4.2.5 SSR-Analyse ........................................................................................................................50 4.2.6 EST-Analyse.........................................................................................................................52 4.3 Statistische Auswertung .............................................................................................................54 4.3.1 Deskriptive Verfahren ...........................................................................................................55 4.3.1.1 Ähnlichkeitsmaß............................................................................................................55 4.3.1.2 Clusteranalyse ..............................................................................................................56 4.3.1.3 Faktorenanalyse ...........................................................................................................57 4.3.2 Populationsgenetische Analysen .........................................................................................57 4.3.2.1 Allelfrequenz .................................................................................................................57 4.3.2.2 Genetische Diversität....................................................................................................58 4.3.2.3 Populationsunterteilung ................................................................................................60 4.3.2.4 Genetische Distanz.......................................................................................................60 4.3.2.5 Genfluss........................................................................................................................61 4.3.2.6 Genetische Variation in Untergruppen .........................................................................61 5 Ergebnisse...........................................................................................................63 5.1 Morphologisch-biometrische Charakterisierung der untersuchten Individuen ...........................63 5.2 Populationsgenetische Analyse der Einzelbäume .....................................................................75 5.2.1 RAPD-PCR...........................................................................................................................75 5.2.1.1 Genetische Ähnlichkeit .................................................................................................77 5.2.1.2 Genetische Distanz und Genfluss ................................................................................82 5.2.1.3 Genetische Diversität....................................................................................................88 5.2.1.4 Zusammenfassung .......................................................................................................91 5.2.2 ISSR-PCR.............................................................................................................................92 5.2.2.1 Genetische Ähnlichkeit .................................................................................................93 5.2.2.2 Genetische Distanz und Genfluss ................................................................................97
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