Dokument

Zusammenfassung:
Ustilago maydis, der Erreger des Maisbeulenbrandes, ist für die Etablierung einer kompatiblen Interaktion mit seiner Wirtspflanze Zea mays auf die Sekretion zahlreicher Effektorproteine angewiesen. Zu solchen Effektoren gehören sekretierte LysM-Proteine phytopathogener Pilze. LysM-Proteine können essentielle Virulenzfaktoren sein, indem sie Chito-Oligosaccharide, welche durch pflanzliche Chitinasen aus der pilzlichen Zellwand freigesetzt wurden, abfangen und dadurch eine PAMP-vermittelte Resistenz der Pflanze unterdrücken. Im Genom von U. maydis wurde ein für ein LysM-Protein kodierendes Gen (um11464) identifiziert und näher untersucht. Die Deletion von um11464 führte zu einem hypervirulenten Phänotyp in Pflanze. Eine detaillierte phänotypische Charakterisierung eines um11464 Deletionsstammes ergab, dass dieser eine veränderte Morphologie in Flüssigkultur sowie eine erhöhte Sensitivität gegenüber Zellwandstressoren aufwies. Ferner wirkte sich die Deletion von um11464 positiv auf die Filament- und Appressorienbildung aus. Die Penetrationseffizienz des Stammes SG200∆um11464 war hingegen im Vergleich zu SG200 unverändert. Sekretionsanalysen zeigten, dass das Signalpeptid von Um11464 funktionell war. Um11464 konnte jedoch zu keinem Zeitpunkt in Kulturüberständen nachgewiesen werden. Durch konfokale Laserscanning-Mikroskopie und Immunmarkierungen wurde gezeigt, dass das Protein in der pilzlichen Zellwand lokalisiert. Es wurde eine C-terminale Verankerung über nicht-kovalente Verknüpfungen nachgewiesen. Eine Interaktion mit Chitin oder anderen pilzlichen Zellwandbestandteilen konnte durch Polysaccharid-Bindeversuche mit rekombinantem Um11464 nicht bestätigt werden. Der positive Effekt der um11464 Deletion auf die Differenzierungsprozesse vor der Penetration sowie die Virulenz von U. maydis könnte andeuten, dass Um11464 eine veränderte Pflanzenreaktion hervorruft. Um dies zu untersuchen, wurden vergleichende Mais-Transkriptomanalysen durchgeführt. Diese zeigten, dass eine Vielzahl abwehrinduzierter Pflanzengene, darunter einige Transkriptionsfaktoren, spezifisch nach Infektion mit der um11464 Deletionsmutante herunterreguliert wurden, was eine aktive Unterdrückung der pflanzlichen Abwehr bedeuten könnte.

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