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Mykologische Und Holzanatomische Untersuchungen Zur Massaria-Krankheit an Platanen

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Mykologische Und Holzanatomische Untersuchungen Zur Massaria-Krankheit an Platanen Mykologische und holzanatomische Untersuchungen zur Massaria-Krankheit an Platanen Inauguraldissertation zur Erlangung der Doktorwürde der Fakultät für Umwelt und Natürliche Ressourcen der Albert-Ludwigs-Universität Freiburg im Breisgau vorgelegt von Timo Börker Freiburg im Breisgau Februar 2019 Dekanin: Prof. Dr. D. Kleinschmit Referent: Prof. Dr. S. Fink Korreferent: Prof. Dr. F. Schwarze Zweiter Betreuer: PD Dr. B. Metzler Datum der Disputation: 06.06.2019 DANKSAGUNG Danksagung Zuerst möchte ich mich bei Herrn Prof. Dr. S. Fink für die Vergabe des Themas bedanken. Insbesondere für die Möglichkeit, die mir geboten wurde, dieses spannende Thema selbstständig mit dem nötigen Freiraum zu bearbeiten. Herrn Prof. Dr. F. Schwarze gilt mein Dank für die freundliche Übernahme des Korreferats und die damit verbundene Arbeit. Einen sehr großen Dank gehört Herrn PD Dr. B. Metzler für die Zweitbetreuung sowie Anregungen und Aufmunterungen mykologischer Fragestellungen während der gesamten Zeit. Besonders danke ich Herrn Prof. Dr. R. Kehr, der bereits während meiner Studienzeit mein Interesse für die Gehölzpathologie weckte. Seine Tipps und Anregungen waren eine große Bereicherung bei der Bearbeitung des Themas. Ganz herzlich möchte ich mich bei Herrn Dr. J. Grüner bedanken. Seine Ratschläge waren eine große Hilfe dieses Thema zu bearbeiten. Als Ansprechpartner stand er mir jederzeit zur Verfügung. Vielen, vielen Dank Jörg! Dem gesamten Team der Forstbotanik danke ich für die freundliche Unterstützung der Arbeiten. Bei Frau Dr. M. Alabed Alkader möchte ich mich für die wertvollen Tipps bei der Vorgehensweise mikrobiologischer Arbeiten bedanken. Frau S. Röske danke ich vor allem für die damit verbundenen Arbeiten der PCR, Frau N. Streit und ganz besonders Frau S. Diener gilt mein Dank für die Unterstützung der Labor- und Feldarbeiten sowie die Anfertigung diverser histologischer Präparate. Herrn R. Schönle gilt mein Dank vor allem bei der Hilfestellung photometrischer Messungen. Ein großer Dank gehört dem Team des Forstbotanischen Gartens, besonders Herrn K. Merz für die Anlage der Feldarbeiten, Herrn R. Dietrich für die Aufbereitung der Holzproben und Herrn W. Jäger für die Wasserversorgung der Versuchsbäume. Bei Frau L. Gschwend möchte ich mich ganz besonders für die nette, angenehme Zeit mit hilfreichen Ratschlägen bedanken. Des Weiteren danke ich meinen Kollegen /Kolleginnen (M. Göttelmann, L. Blanquer & K. Drozella) für das tolle Arbeitsklima in dieser Zeit. Herrn U. Uhlich (Professur für Forstliche Biomaterialien) gilt mein Dank für die Möglichkeit der Druckfestigkeitsprüfung. Dem Garten und Tiefbauamt der Stadt Freiburg sowie den Baumpartnern-Breisgau danke ich für das Probematerial diverser Äste. Abschließend möchte ich mich ganz herzlich bei meiner gesamten Familie und meinen Freunden bedanken, die mich jederzeit unterstützt haben und mir während der gesamten Promotion Rückhalt gaben. Ohne diese Unterstützung wäre dies alles nicht möglich gewesen. Vielen, vielen Dank für Alles. INHALTSVERZEICHNIS Inhaltsverzeichnis 1. Einleitung ............................................................................................................... 1 1.1 Die Gattung Platanus .................................................................................. 2 1.1.1 Platanen im urbanen Raum ..................................................................... 3 1.2 Stand des Wissens zur „Massaria-Krankheit“ ........................................... 4 1.2.1 Krankheitsverlauf und Symptome der „Massaria-Krankheit“ .............. 5 1.2.2 Holzanatomie von Platanen .................................................................... 7 1.2.3 Fäuletypen holzzersetzender Pilze .......................................................... 8 1.3 Zielsetzung der Arbeit ................................................................................. 9 2. Material und Methoden ........................................................................................ 11 2.1 Pilzmaterial .................................................................................................. 11 2.1.1 Isolierung und Kultivierung von Splanchnonema platani ...................... 11 2.1.2 Pilzidentifikation ..................................................................................... 12 2.2 Nährmedien .................................................................................................. 15 2.2.1 Nährmedien mit Holz- bzw. Rindenzusatz ............................................. 17 2.3 Wachstumsversuche ..................................................................................... 18 2.3.1 Temperaturoptimum ............................................................................... 18 2.3.2 Substratansprüche ................................................................................... 18 2.4 Holzabbau .................................................................................................... 19 2.4.1 Herstellung der Prüfkörper ..................................................................... 19 2.4.2 Verwendete Pilzkulturen ......................................................................... 20 2.4.3 Vorgehensweise zur Anzucht der Pilze und Beimpfung der Versuchsgefäße ....................................................................................... 21 2.4.4 Druckfestigkeitsprüfung an pilz-infizierten Platanenhölzern ................ 23 2.5 Herstellung histologischer Präparate ........................................................... 24 2.5.1 Färbungen ............................................................................................... 25 2.6 Nachweis qualitativer Enzymaktivität ........................................................ 26 2.6.1 Dualkultur-Test zum antagonistischen Potential von Trichoderma sp. .. 27 2.7 Holzanatomische Strukturen an Platanenästen ............................................ 29 2.8 Infektionsversuch an Bäumen bei unterschiedlicher Wasserversorgung .... 30 2.8.1 Versuchsfläche im Forstbotanischen Garten .......................................... 30 2.8.2 Versuchsaufbau ....................................................................................... 30 2.8.3 Herstellung des Inokulums ..................................................................... 35 2.8.4 Isolation von Mikroorganismen aus Rindengewebe der Platanenäste............................................................................................. 36 2.8.5 Wetterdaten ............................................................................................. 37 2.9 Statistische Auswertung ............................................................................... 37 3. Ergebnisse .............................................................................................................. 38 3.1 Molekulargenetische Bestimmung ............................................................... 38 3.2 Kultivierung .................................................................................................. 40 3.3 Wachstumsversuche ..................................................................................... 41 3.3.1 Temperaturoptimum ............................................................................... 41 INHALTSVERZEICHNIS 3.3.2 Substratansprüche ................................................................................... 45 3.4 Holzabbau ................................................................................................... 50 3.4.1 Holzfeuchte der Platanenhölzer ............................................................... 55 3.4.2 Beziehung zwischen Gewichtsverlust und Holzfeuchte.......................... 58 3.4.3 Druckfestigkeitsmessungen an pilz-infizierten Hölzern ......................... 60 3.4.4 Durchlichtmikroskopische Untersuchungen über das Besiedlungs- und Holzzersetzungsmuster in vitro ............................................................... 63 3.4.5 Fluoreszenzmikroskopische Untersuchungen der Zellwände ................. 72 3.5 Ergebnisse der qualitativen Enzymaktivität ................................................ 74 3.5.1 Dualkultur-Test zur Ermittlung des antagonistischen Potentials von Trichoderma sp. ....................................................................................... 75 3.6 Zugholz in Platanenastgewebe .................................................................... 80 3.6.1 Makroskopische Auswertung .................................................................. 80 3.6.2 Mikroskopische Auswertung ................................................................... 80 3.7 Ergebnisse der Infektionsversuche (in vivo) ................................................ 83 3.7.1 Wetterdaten.............................................................................................. 83 3.7.2 Zuwachsmessungen an den Versuchsbäumen ......................................... 84 3.7.3 Infektionsversuch I Erreger auf intaktem Rindenperiderm ..................... 86 3.7.4 Astbasis als natürliche Barriere ............................................................... 93 3.7.5 Infektionsversuch II Erreger auf geschädigtem Rindenperiderm ............ 95 3.7.6 Isolation aus Rindengewebe von Platanenästen ...................................... 100 3.7.7 Gemeinsames Auftreten von S. platani und Phomopsis/ Diaporthe spp. im Rindengewebe ................................................................................... 104 3.8 Holz-Beimpfung mit Pilzen aus Platanenrindengewebe in vitro ................
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