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Untersuchungen Zur Regulation Der Polyphenolbiosynthese in Der Erdbeerfrucht (Fragaria  Ananassa) Mittels Metabolite Profiling

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Untersuchungen Zur Regulation Der Polyphenolbiosynthese in Der Erdbeerfrucht (Fragaria  Ananassa) Mittels Metabolite Profiling TECHNISCHE UNIVERSITÄT MÜNCHEN Fachgebiet Biotechnologie der Naturstoffe Untersuchungen zur Regulation der Polyphenolbiosynthese in der Erdbeerfrucht (Fragaria ananassa) mittels Metabolite Profiling Ludwig F. M. Ring Vollständiger Abdruck der von der Fakultät Wissenschaftszentrum Weihenstephan für Ernährung, Landnutzung und Umwelt der Technischen Universität München zur Erlangung des akademischen Grades eines Doktors der Naturwissenschaften genehmigten Dissertation. Vorsitzende: Univ.-Prof. Dr. B. Poppenberger Prüfer der Dissertation: 1. Univ.-Prof. Dr. W. Schwab 2. Univ.-Prof. Dr. Th. Hofmann 3. Univ.-Prof. Dr. D. R. Treutter Die Dissertation wurde am 17.06.2013 bei der Technischen Universität München eingereicht und durch die Fakultät Wissenschaftszentrum Weihenstephan für Ernährung, Landnutzung und Umwelt am 22.10.2013 angenommen. „… and all the pieces matter“ Lester Freamon, 2002 meiner Familie Danksagung I Danksagung Meinem Doktorvater Prof. Dr. Wilfried Schwab gilt mein besonderer Dank für die Überlassung des Themas und die Möglichkeit an seinem Fachgebiet zu promovieren. Außerdem danke ich ihm für seine immerwährende Unterstützung und seinen ausstrahlenden Optimismus. Bei Prof. Dr. Brigitte Poppenberger, Prof. Dr. Thomas Hofmann und Prof. Dr. Dieter Treutter bedanke ich mich für die Mitarbeit in der Prüfungskommission. Allen Kooperationspartnern des FraGenomics-Projekts, insbesondere Prof. Dr. Juan Muñoz- Blanco, Dr. Beatrice Denoyes-Rothan und Dr. Amparo Monfort, möchte ich für die gute Zusammenarbeit und die fruchtbaren Diskussionen bei den Projekttreffen danken. Prof. Dr. Victoriano Valpuesta danke ich sehr für die Möglichkeit meine Arbeiten zur Proteinanalytik am Department für Molekularbiologie und Biochemie der Universität Málaga durchführen zu können. Seinem gesamten Arbeitskreis danke ich für die herzliche Aufnahme! Gracias a todos los miembros del grupo! Además, les agradesco a Dra. Irene Aragüez-Rey y Paqui Martínez sus ayudas con los geles 2D, y gracias a Ana Casañal y Dra. Cristina Muñoz por el apoyo local estupendo! Bei Dr. Thomas Hoffmann und Dr. Anja Preuß möchte ich mich sehr für die gewissenhafte Durchsicht meines Manuskripts und die wertvollen Denkanstöße bedanken. Dr. Stephanie Hücherig und Dr. Su-Ying Yeh danke ich für die weiterführenden Arbeiten an FaPRX27. Dipl.-LMChem. Ingolf Krause (†) verdanke ich einen Großteil meines LC-MS-Wissens, weshalb ich ihm für die zahlreichen „Lehrstunden“ zur LC-MS danken will. Allen aktuellen und ehemaligen Mitarbeitern des BiNa-Fachgebiets gilt mein herzlicher Dank für die stetige Hilfsbereitschaft und das angenehme Arbeitsklima. Im Speziellen danke ich Quirin Sinz und Christopher Fuchs für die hervorragende Atmosphäre im gemeinsamen Büro. Bei Corina Sutter und Christine John bedanke ich mich für ihre äußerst engagierte Mitarbeit im Rahmen von Masterarbeit und Forschungspraktikum. Der größte Dank gebührt meinen Eltern, da sie mir diesen Schritt und vieles mehr überhaupt erst möglich machten. Auf ihre bedingungslose Unterstützung kann ich immer vertrauen. Genauso danke ich Hanna für den starken Rückhalt, die Motivationskraft und ihre unendliche Geduld. II Publikationen Publikationen Die Arbeiten zur vorliegenden Dissertation wurden im Zeitraum Mai 2009 bis Mai 2012 in der Arbeitsgruppe Biotechnologie der Naturstoffe (früher Biomolekulare Lebensmitteltechnologie) am Wissenschaftszentrum Weihenstephan der Technischen Universität München in Freising durchgeführt. Die Dissertation entstand im Rahmen des transnationalen PLANT-KBBE- Verbundprojekts „FraGenomics: Genetical genomics for improving strawberry fruit nutritional quality“, gefördert durch das Bundesministerium für Bildung und Forschung (BMBF). Auszüge aus dieser Dissertation wurden bereits zur Veröffentlichung eingereicht bzw. veröffentlicht. Originalartikel: Ring L, Yeh S-Y, Hücherig S, Hoffmann T, Blanco-Portales R, Fouche M, Villatoro C, Denoyes B, Monfort A, Muñoz-Blanco J, Gershenson J, Schwab W (2013) Metabolic interaction between anthocyanin and lignin biosynthesis is associated with a peroxidase in strawberry fruit. Plant Physiology 163(1): 43-60. Fischer T C, Mirbeth B, Rentsch J, Sutter C, Ring L, Flachowsky H, Habegger R, Hoffmann T, Hanke M-V, Schwab W (2013) Premature and ectopic anthocyanin formation by silencing of anthocyanidin reductase in strawberry (Fragaria ananassa). New Phytologist doi: 10.1111/nph.12528. Cumplido-Laso G, Medina-Puche L, Amil-Ruiz F, Moreno-Suarez B, Ring L, Rodriguez- Franco A, Caballero JL, Schwab W, Muñoz-Blanco J, Blanco-Protales R (2013) FaMYB10 plays a major role in the regulation of the flavonoid/phenylpropanoid metabolism during the ripening of Fragaria ananassa fruits. Journal of Experimental Botany doi: 10.1093/jxb/ert377. Cumplido-Laso G, Medina-Puche L, Moyano E, Hoffmann T, Sinz Q, Ring L, Studart- Witkowski C, Caballero JL, Schwab W, Munoz-Blanco J, Blanco-Portales R (2012) The fruit ripening-related gene FaAAT2 encodes an acyl transferase involved in strawberry aroma biogenesis. Journal of Experimental Botany 63(11): 4275-4290. Habegger R, Ring L, Schwab W (2012) Erdbeeren mit gesteigertem Gesundheitswert: Potenzial für Züchtung. Spargel & Erdbeerprofi 14(5):74. Schwab W, Preuß A, Ring L (2010) Gesunde Früchtchen. GenomXpress 2:25-57. Publikationen III Vorträge: Schwab W, Ring L, Preuß A (2010) FraGenomics: Genetical Genomics for Improving Strawberry Fruit Nutritional Quality (Stand 2010). GABI Statusseminar 2010, Potsdam. Schwab W, Ring L, Preuß A (2011) FraGenomics: Genetical Genomics for Improving Strawberry Fruit Nutritional Quality (Stand 2011). GABI Statusseminar 2011, Potsdam. Fischer T, Ring L, Hoffmann T, Sutter C, Mirbeth B, Hanke V, Flachowsky H, Schwab W (2011) Silencing of anthocyanidin reductase in strawberry (Fragaria ananassa). Botaniker Tagung, Berlin. Ring L, Schwab W (2012) Metabolite Profiling combined with transcriptome analyses reveal novel candidate genes involved in polyphenol biosynthesis. PLANT-KBBE final seminar, Barcelona. Posterpräsentationen: Ring L, Preuß A, Schwab W (2010) FraGenomics: Genetical Genomics for Improving Strawberry Fruit Nutritional Quality (Stand 2010). GABI Statusseminar 2010, Potsdam. Schwab W, Ring L, Preuß A (2011) FraGenomics: Genetical Genomics for Improving Strawberry Fruit Nutritional Quality (Stand Anfang 2011). GABI Statusseminar 2011, Potsdam. Ring L, Schwab W (2011) FraGenomics: Genetical Genomics for Improving Strawberry Fruit Nutritional Quality (Stand Ende 2011). Max-Rubner Conference on Food Metabolomics, Karlsruhe. Fouche M, Vignes M, Mangin B, Petit A, Chartier P, Schwab W, Ring L, Monfort A, Villatoro C, Rothan C, Denoyes B (2012) Role of a MYB factor in strawberry fruit color. 6th Rosaceous Genomics Conference, Trient. Schwab W, Ring L, Hücherig S, Yeh SY, Preuß A (2012) Metabolite profiling and transcriptome analyses reveal novel candidate genes. PLANT2030 GABI Statusseminar 2012, Potsdam. Ring L, Muñoz-Blanco J, Schwab W (2012) PLANT-KBBE Fragenomics: Metabolite profiling of strawberry fruit. PLANT2030 GABI Statusseminar 2012, Potsdam. IV Inhaltsverzeichnis Inhaltsverzeichnis Abkürzungen .................................................................................................. IX Strukturformeln ............................................................................................ XIV Zusammenfassung ..................................................................................... XVII Summary ........................................................................................................ XX 1. Einleitung ............................................................................................. 1 1.1. Erdbeere ............................................................................................................... 1 1.1.1. Geschichte ....................................................................................................................................... 1 1.1.2. Morphologie .................................................................................................................................... 2 1.1.3. Reifung ............................................................................................................................................. 3 1.1.4. Produktion und Konsum .............................................................................................................. 4 1.1.5. Inhaltsstoffe .................................................................................................................................... 5 1.2. Polyphenole ......................................................................................................... 7 1.2.1. Biosynthese und Funktionen in planta ........................................................................................ 7 1.2.2. Ernährungsphysiologische Aspekte .......................................................................................... 10 1.2.3. Ligninbiosynthese ........................................................................................................................ 11 1.3. Konzepte der Metabolit-Analytik ........................................................................ 12 1.4. Aufgabenstellung ................................................................................................ 14 2. Material und Methoden ....................................................................... 16 2.1. Material ..............................................................................................................
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