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Methodenentwicklung Zur Herstellung Cisgener Apfelpflanzen

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Methodenentwicklung Zur Herstellung Cisgener Apfelpflanzen Juliane Würdig Institut für Züchtungsforschung an Obst Methodenentwicklung zur Herstellung cisgener Apfelpfl anzen Dissertationen aus dem Julius Kühn-Institut Julius Kühn-Institut Bundesforschungsinstitut für Kulturpfl anzen Kontakt/Contact: Juliane Schiffler Julius Kühn-Institut (JKI) Bundesforschungsinstitut für Kulturpflanzen Institut für Züchtungsforschung an Obst Pillnitzer Platz 3a 01326 Dresden Die Schriftenreihe ,,Dissertationen aus dem Julius Kühn-Institut" veröffentlicht Doktorarbeiten, die in enger Zusammenarbeit mit Universitäten an Instituten des Julius Kühn-Instituts entstanden sind. The publication series „Dissertationen aus dem Julius Kühn-Institut" publishes doctoral dissertations originating from research doctorates and completed at the Julius Kühn-Institut (JKI) either in close collaboration with universities or as an outstanding independent work in the JKI research fields. Der Vertrieb dieser Monographien erfolgt über den Buchhandel (Nachweis im Verzeichnis lieferbarer Bücher - VLB) und OPEN ACCESS im Internetangebot www.julius-kuehn.de Bereich Veröffentlichungen. The monographs are distributed through the book trade (listed in German Books in Print - VLB) and OPEN ACCESS through the JKI website www.julius-kuehn.de (see Publications). Wir unterstützen den offenen Zugang zu wissenschaftlichem Wissen. Die Dissertationen aus dem Julius Kühn-lnstitut erscheinen daher OPEN ACCESS. Alle Ausgaben stehen kostenfrei im Internet zur Verfügung: http://www.julius-kuehn.de Bereich Veröffentlichungen. We advocate open access to scientific knowledge. Dissertations from the Julius Kühn-Institut are therefore published open access. All issues are available free of charge under http://www.julius-kuehn.de (see Publications). Bibliografische Information der Deutschen Nationalbibliothek Die Deutsche Nationalbibliothek verzeichnet diese Publikation In der Deutschen Nationalbibliografie: detaillierte bibliografische Daten sind im Internet über http://dnb.d-nb.de abrufbar. Bibliographic information published by the Deutsche Nationalbibliothek (German National Library) The Deutsche Nationalbibliothek lists this publication in the Deutsche Nationalbibliografie; detailed bibliographic data are available in the Internet at http://dnb.dnb.de. ISBN 978-3-95547-048-7 DOI 10.5073/dissjki.2017.006 Herausgeber l Editor Julius Kühn-Institut, Bundesforschungsinstitut für Kulturpflanzen, Quedlinburg, Deutschland Julius Kühn-Institut, Federal Research Centre for Cultivated Plants, Quedlinburg, Germany Dieses Werk ist lizenziert unter einer Creative Commons – Namensnennung – Weitergabe unter gleichen Bedingungen – 4.0 Lizenz. This work is licensed under a Creative Commons – Attribution – ShareAlike – 4.0 license. Methodenentwicklung zur Herstellung cisgener Apfelpflanzen DISSERTATION zur Erlangung des akademischen Grades Doctor rerum naturalium (Dr. rer. nat.) vorgelegt der Fakultät Mathematik und Naturwissenschaften der Technischen Universität Dresden von Diplom-Biologin Juliane Würdig geboren am 03. April 1984 in Berlin Die Dissertation wurde in der Zeit von März / 2011 bis März / 2016 am Julius Kühn-Institut (JKI), im Institut für Züchtungsforschung an Obst (ZO) in Dresden-Pillnitz angefertigt. Eingereicht am 13. April 2016 Angenommen am 08. Februar 2017 1. Gutachter (schriftliche Beurteilung): Frau Prof. Dr. Jutta Ludwig-Müller 2. Gutachter (schriftliche Beurteilung): Frau Prof. Dr. Magda-Viola Hanke Tag der Disputation: 11. April 2017 Promotionskommission: Herr Prof. Dr. Michael Rother (Vorsitzender) Frau Prof. Dr. Jutta Ludwig-Müller Frau Prof. Dr. Magda-Viola Hanke Herr Prof. Dr. Michael Göttfert „Der Weg ist das Ziel.“ (Konfuzius) Versicherung Versicherung Hiermit versichere ich, dass ich die vorliegende Arbeit ohne unzulässige Hilfe Dritter und ohne Benutzung anderer als der angegebenen Hilfsmittel angefertigt habe. Die aus fremden Quellen direkt oder indirekt übernommenen Gedanken sind als solche kenntlich gemacht. Die Arbeit wurde bisher weder im Inland noch im Ausland in gleicher oder ähnlicher Form einer anderen Prüfungsbehörde vorgelegt. Datum, Unterschrift 5 Inhaltsverzeichnis Inhaltsverzeichnis VERSICHERUNG .................................................................................................. 5 INHALTSVERZEICHNIS ....................................................................................... 6 ZUSAMMENFASSUNG ...................................................................................... 11 ABSTRACT ......................................................................................................... 13 PUBLIZIERTE ERGEBNISSE AUS DER VORLIEGENDEN DISSERTATION .. 15 ABKÜRZUNGSVERZEICHNIS ........................................................................... 16 1 EINLEITUNG ................................................................................................ 19 1.1 DER APFEL ............................................................................................. 19 1.1.1 Systematik und Taxonomie ......................................................................... 19 1.1.2 Herkunft und Bedeutung als Kultursorte ..................................................... 19 1.2 DER APFELSCHORF (VENTURIA INAEQUALIS) ............................................. 20 1.2.1 Probleme im Apfelanbau............................................................................. 20 1.2.2 Systematik und Taxonomie ......................................................................... 21 1.2.3 Lebenszyklus .............................................................................................. 21 1.2.4 Symptome .................................................................................................. 22 1.3 NATÜRLICHE APFELSCHORFRESISTENZEN VON MALUS .............................. 22 1.3.1 Überblick .................................................................................................... 22 1.3.2 Der Schorfresistenzlocus Rvi6 .................................................................... 23 1.3.3 Der Schorfresistenzlocus Rvi15 .................................................................. 25 1.4 DAS PATHOSYSTEM V. INAEQUALIS-MALUS ............................................... 26 1.4.1 Gen-für-Gen Beziehung .............................................................................. 26 1.4.2 V. inaequalis Rassen .................................................................................. 26 1.4.3 Zusammenbruch der Rvi6 Schorfresistenz ................................................. 26 1.5 DIE APFELZÜCHTUNG .............................................................................. 27 1.5.1 Überblick .................................................................................................... 27 1.5.2 Die Züchtung schorfresistenter Sorten ........................................................ 28 1.5.3 Gegenwärtige Ziele ..................................................................................... 28 1.5.4 Gentechnik in der Apfelzüchtung ................................................................ 29 1.6 DAS KONZEPT DER CISGENETIK ............................................................... 29 1.6.1 Einleitung .................................................................................................... 29 1.6.2 Definition und Umsetzung beim Apfel ......................................................... 30 1.6.3 Clean Vector Technologie-Vektoren in der Apfelzüchtung .......................... 31 6 Inhaltsverzeichnis 1.6.3.1 Das Dexamethason induzierbare R/Rs Rekombinasesystem .......................... 32 1.6.3.2 Das hitzeinduzierbare Flp/FRT Rekombinasesystem ...................................... 33 1.7 MYB10 (R6) ALS ALTERNATIVE ZUM REKOMBINASESYSTEM FÜR DEN CISGENETISCHEN ANSATZ BEIM APFEL ................................................................ 34 1.7.1 Einleitung .................................................................................................... 34 1.7.2 MYB Transkriptionsfaktoren in Pflanzen ..................................................... 35 1.7.3 Anthocyane in Pflanzen .............................................................................. 36 1.7.4 Regulation der Anthocyanbiosynthese in Pflanzen ..................................... 36 1.7.4.1 Transkriptionelle Steuerung der Anthocyanbiosynthese im Apfel .................... 36 1.7.5 Anthocyananreicherung beim Apfel ............................................................ 37 1.7.5.1 Typ 1 für Rotfleischigkeit beim Apfel ................................................................ 37 1.7.5.2 Typ 2 für Rotfleischigkeit beim Apfel ................................................................ 38 1.8 ZIELSTELLUNG ........................................................................................ 39 2 MATERIAL UND METHODEN ..................................................................... 41 2.1 MATERIAL ............................................................................................... 41 2.1.1 Puffer und Lösungen .................................................................................. 41 2.1.2 Medien ........................................................................................................ 42 2.1.3 Verwendete Programme ............................................................................. 43 2.1.4 Verwendete binäre Vektoren ...................................................................... 43 2.1.4.1 pB-Npt-Hsp-Flp-Gus ........................................................................................
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