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Nachweis der GattungArmillaria (Fr.:Fr.) Staude undHeterobasidion annosum (Fr.) Bref. in Fichte (Picea abies [L.] Karst.) und Erfassung der klonalen Ausbreitung vonA. ostoyae-Genotypen unter Verwendung molekularer Methoden

Identification of the genusArmillaria (Fr.:Fr.) Staude andHeterobasidion annosum (Fr.) Bref. in Norway spruce (Picea abies [L.] Karst.) and determination of clonal distribution ofA. ostoyae-genotypes by molecular methods

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Zusammenfassung

Methoden zur Entwicklung molekularer Marker mit Hilfe der Polymerase-Kettenreaktion (PCR) werden beispielhaft an den beiden forstlichen PathogenenArmillaria spp. undH. annosum erläutert. Unter Verwendung der aus „internal transcribed spacer“ (ITS)-DNA-Sequenzen abgeleiteten Primerpaare ARM-1/ARM-2 bzw. HET-7/HET-8 konntenArmillaria spp.- bzw.H. annosum-spezifische DNA-Abschnitte mittels PCR amplifiziert werden. Diese PCR-Diagnosemethode erlaubt einen schnellen und effizienten Frühnachweis dieser beiden forstlich relevanten Wurzel- und Stockfäuleerreger in verschiedenen Substraten bzw. Pflanzengeweben. Die genetische Variabilität von 20A. ostoyae-Isolaten aus verschiedenen geographischen Herkunftsgebieten wurde untersucht. Die UPGMA-Clusteranalyse der unter Verwendung von 10 Decamer-Zufallsprimern (OPA 01-10) erhaltenen „random amplified polymorphic DNAs“ (RAPDs)-Muster gruppierte die Isolate in Untergruppen mit 40–96% Ähnlichkeit, was auf eine hohe intraspezifische genetische Variation hindeutet. Die potentielle Rolle der historischen und gegenwärtigen Ausbreitung von Fichtenpflanzen für die genetische Variation vonA. ostoyae in Europa wird diskutiert. Die aufgefundenen polymorphen DNA-Marker wurden für die Erfassung der Populationsstruktur bzw. -dynamik sowie der räumlichen Ausbreitung vonA. ostoyae-„genets“ in ausgewählten Befallsgebieten genutzt. Erste Untersuchungen haben unterschiedliche Ausbreitungsstrategien vonA. ostoyae in Abhängigkeit von Klima-, Standort- und Immissionsfaktoren aufgezeigt.

Summary

Methods based on the polymerase chain reaction (PCR) for generating molecular markers are illustrated by two important forest pathogens,Armillaria spp. andH. annosum. Using the primer pairs ARM-1/ARM-2 and HET-7/HET-8, derived from sequences of the “internal transcribed spacer” (ITS) regions of the rDNA repeat,Armillaria spp.- andH. annosum-specific DNA fragments were amplified by PCR. This PCR-based detection method allows a rapid and definite diagnosis of both important root and butt rot pathogens in different substrates or plant tissues, especially in early stages. Genetic variability among 20A. ostoyae-isolates from different geographical origins was studied. UPGMA cluster analysis of random amplified polymorphic DNAs (RAPDs) profiles generated by 10 decamer random primers (OPA 01-10) grouped the isolates in subclusters at similarity levels between 40% and 96%, indicating high intraspecific genetic variation. The potential role of historical and current spread of spruce plants on the genetic variation ofA. ostoyae in Europe is discussed. The established polymorphic DNA markers were used to determine the population structure, dynamics and spatial distribution ofA. ostoyae-“genets” in areas colonized by the fungus. First investigations revealed different distribution strategies ofA. ostoyae, which may be mediated by climatic factors, location (e.g., soil characters), and pollutants.

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Authors and Affiliations

  1. Institut für Forstbotanik und Forstzoologie, TU Dresden, PF 10, D-01735, Tharandt

    S. Schulze

  2. GSF-Forschungszentrum für Umwelt und Gesundheit, Institut für Biochemische Pflanzenpathologie, PF 1129, D-85758, Oberschleißheim

    G. Bahnweg

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  1. S. Schulze
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  2. G. Bahnweg
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Schulze, S., Bahnweg, G. Nachweis der GattungArmillaria (Fr.:Fr.) Staude undHeterobasidion annosum (Fr.) Bref. in Fichte (Picea abies [L.] Karst.) und Erfassung der klonalen Ausbreitung vonA. ostoyae-Genotypen unter Verwendung molekularer Methoden. Forstw Cbl 117, 98–114 (1998). https://doi.org/10.1007/BF02832963

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