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Untersuchungen Zur Vermehrung Und Mykorrhiza Von Eleutherococcus Maxim

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Untersuchungen Zur Vermehrung Und Mykorrhiza Von Eleutherococcus Maxim Untersuchungen zur Vermehrung und Mykorrhiza von Eleutherococcus Maxim. (Araliaceae) Dissertation zur Erlangung des Doktorgrades der Naturwissenschaften (Dr. rer. nat.) dem Fachbereich Biologie der Philipps-Universität Marburg vorgelegt von Matthias Döring aus Meißen (Freistaat Sachsen) Marburg/Lahn 2007 Vom Fachbereich Biologie der Philipps-Universität Marburg als Dissertation am 07.02.2007 angenommen. Erstgutachter: Prof. Dr. Hans Christian Weber Zweitgutachter: Prof. Dr. Roland Brandl Tag der mündlichen Prüfung am 08.02.2007. ii Meinen lieben Eltern gewidmet iii INHALTSVERZEICHNIS 1. Einleitung.......................................................................................................................1 1.1. Name.................................................................................................................................1 1.2. Verbreitung und Artenschutz............................................................................................2 1.3. Morphologie und Anatomie .............................................................................................4 1.4. Nutzen...............................................................................................................................5 1.5. Kultivierung und Vermehrung .......................................................................................10 1.6. Mykorrhiza .....................................................................................................................12 1.7. Zielsetzung .....................................................................................................................26 2. Material und Methoden ............................................................................................27 2.1. Untersuchtes Pflanzenmaterial.......................................................................................27 2.2. Entnahme, Fixierung und Lagerung der Wurzelproben .................................................27 2.3. Anatomische Untersuchungsmethoden ..........................................................................28 2.3.1. Lichtmikroskopie............................................................................................................28 2.3.2. Trypanblaufärbung .........................................................................................................28 2.3.3. Herstellung von Semidünnschnitten in Technovit..........................................................28 2.3.4. Herstellung von Paraffinschnitten..................................................................................29 2.3.5. Nachweis verschiedener Inhaltsstoffe ............................................................................30 2.3.6. Nachweis der Zellkerne..................................................................................................30 2.3.7. Nachweis von aktiv bewegenden Mikroorganismen......................................................30 2.3.8. Rasterelektronmikroskopie (REM)................................................................................30 2.4. Kultivierung von Laubblättern und Sprossstecklingen ..................................................30 2.4.1. Bewurzelung von Laubblättern und Fiederblättchen......................................................30 2.4.2. Behandlungen mit Rotlicht.............................................................................................31 2.5. Etablierung einer Pflanzengewebekultur von Eleutherococcus sieboldianus (Makino) Koidz. .............................................................................................................31 2.6. Isolation und Kultivierung von Mikroorganismen .........................................................32 2.7. Topfkultivierung und Inokulation mit Glomus intraradices in vivo ..............................33 2.7.1. Inokulumgewinnung.......................................................................................................33 2.7.2. Inokulationsexperiment mit Eleutherococcus sieboldianus...........................................33 2.7.3. Inokulationsexperiment mit Eleutherococcus gracilistylus ...........................................33 2.8. Kultur von Gigaspora margarita und Inokulation in vitro ............................................34 2.8.1 Inokulumgewinnung.......................................................................................................34 2.8.2. Desinfektion ...................................................................................................................34 2.8.3. In vitro Kulturmedien für Gigaspora margarita............................................................34 2.8.4. Inokulation von in vitro vermehrten Eleutherococcus sieboldianus mit Gigaspora margarita ........................................................................................................................34 2.9. Wachstumsparameter......................................................................................................35 2.10. Statistik...........................................................................................................................35 2.11. Molekularbiologische Bestimmung der Glomeromyzeten an Wurzeln aus dem Frei- land .................................................................................................................................35 3. Ergebnisse....................................................................................................................38 3.1. Morphologische Merkmale von verschiedenen Eleutherococcus-Arten........................38 3.2. Bewurzelungsexperimente mit Laubblättern von Eleutherococcus gracilistylus ..........53 3.3. Bewurzelungsexperimente mit Laubblättern von verschiedenen Eleutherococcus- Arten ...............................................................................................................................57 3.4. Ausbildung von Adventivsprossen bei verschiedenen Eleutherococcus-Arten.............64 iv 3.5. Stecklingsvermehrung von Sprossachsen verschiedener Eleutherococcus-Arten .........67 3.6. Pflanzengewebekultur von Eleutherococcus sieboldianus.............................................71 3.6.1. Somatische Embryoidogenese von Eleutherococcus sieboldianus................................71 3.6.2. Deformationen des Pflanzenmaterials in der Pflanzengewebekultur.............................80 3.6.3. Mikroorganismen in der Pflanzengewebekultur.............................................................83 3.7. Wurzelanatomie bei Eleutherococcus ............................................................................90 3.8. Mykorrhiza bei Wurzeln aus dem Freiland und Adventivwurzeln von Blattrisslingen von Eleutherococcus gracilistylus..................................................................................94 3.9. Molekularbiologische Untersuchungen der mit Eleutherococcus-Arten assoziierten Glomeromyzeten.............................................................................................................98 3.10. Experimente mit Sporen von Gigaspora margarita Becker & Hall ............................100 3.11. Experiment zum Gravitropismus bei Gigaspora margarita ........................................107 3.12. Inokulation von mikrovermehrten Pflanzen der Art Eleutherococcus sieboldianus mit Gigaspora margarita..............................................................................................109 3.13. Inokulationsexperiment mit Glomus intraradices bei mikrovermehrten Pflanzen von Eleutherococcus sieboldianus ...............................................................................111 4. Diskussion .................................................................................................................117 4.1. Morphologische Beschreibungen der Sträucher...........................................................117 4.2. Anatomische Beschreibungen der Wurzeln .................................................................117 4.3. Bewurzelung von Laubblättern und Stecklingsvermehrung verschiedener Eleutherococcus-Arten .................................................................................................117 4.4. Somatische Embryoidogenese von Eleutherococcus sieboldianus..............................120 4.5. Deformationen von Wurzelhaaren und Pflanzen und Mikroorganismen assoziiert mit Pflanzen von Eleutherococcus sieboldianus aus Pflanzengewebekultur ...............121 4.6. Mykorrhiza von Eleutherococcus.................................................................................125 4.7. Molekularbiologische Charakterisierung der Glomeromyzeten an Wurzeln von verschiedenen Eleutherococcus-Arten .........................................................................131 4.8. In vitro Experimente mit Gigaspora margarita ...........................................................134 4.9. Inokulationsexperiment mit Eleutherococcus sieboldianus und Glomus intraradices 137 5. Anhang .......................................................................................................................141 5.1. Anhang A - Drehbuch ..................................................................................................141 5.2. Anhang B – Inokulationsexperiment von Eleutherococcus sieboldianus mit Glomus intraradices...................................................................................................................151 6. Zusammenfassung ...................................................................................................153
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