Zusammenfassung:
Ustilago maydis, der Erreger des Maisbeulenbrandes, benötigt zur Etablierung einer biotrophen Interaktion mit seiner Wirtspflanze Mais eine Vielzahl sekretierter Effektoren. Die Deletion des für den Effektor Pep1 kodierenden Gens um01987 führt zum vollständigen Pathogenitätsverlust. Die Pflanze reagiert auf Penetrationsversuche der Δpep1-Mutante mit der Induktion verschiedener Abwehrreaktionen, was sich z.B. durch die Bildung nekrotischer Läsionen sowie reaktiver Sauerstoffspezies an den Penetrationsstellen zeigt. Pep1 ist zwischen verschiedenen Brandpilzarten konserviert und besteht aus einem N-terminalen Signalpeptid, einem konservierten Kernbereich sowie einem weniger konservierten Glycin- und Serin-reichen C-Terminus. In der vorliegenden Arbeit wurde das heterolog in E. coli exprimierte Pep1 aufgereinigt. Durch biochemische und molekularbiologische Methoden konnten zwei pflanzliche Bindungspartner von Pep1 identifiziert. So konnte mittels Aktivitäts-basiertem Protein-Profiling gezeigt werden, dass Pep1 spezifisch apoplastische Papain-ähnliche Cysteinproteasen inhibiert. Diese Proteasen spielen offenbar eine wichtige Rolle in der Abwehrinduktion in der Maispflanze.
Des Weiteren bindet und inaktiviert Pep1 pflanzliche Klasse-III-Häm-Peroxidasen. In Kooperation mit Christoph Hemetsberger konnte die Hemmung der Horseradish-Peroxidase (HRP) durch Pep1 in vitro gezeigt. Außerdem konnte die physische Interaktion von Pep1 mit der HRP bewiesen werden. Während der Δpep1-Infektion wird eine Mais Peroxidase, die sekretierte Peroxidase-12, transkriptionell induziert. Durch Virus-induziertes-Gen-silencing der Peroxidase-12 konnte eine partielle Restauration des Δpep1-Phänotyps auf POX12si-Pflanzen bewirkt werden. So führte das silencing der Peroxidase-12 zu einer Unterdrückung pflanzlicher Abwehrreaktionen, wie z.B. der Papillenbildung, was eine erfolgreiche Penetration der Δpep1-Mutante ermöglichte.
Zusammenfassend zeigen diese Ergebnisse, dass Pep1 Enzyme der basalen Pflanzenabwehr inhibiert und somit die angeborene Resistenz der Wirtspflanze gegen U. maydis unterdrückt.
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