Abstract
The paper describes genetic studies on house rot fungi and a rapid identification technique for them, viz. the true dry rot fungus Serpula lacrymans, the wild merulius S. himantioides, the cellar fungus Coniophora puteana and the pore house fungi Antrodia vaillantii and Tyromyces placenta. The method ARDRA-ITS uses the internal transcribed spacer (ITS) of the ribosomal DNA (rDNA) which is amplified by the polymerase chain reaction (PCR). Amplifications were directly performed from agar plate mycelium and even from fungal tissue taken from infected substrate in a building without preceding fungal pure cultures and DNA preparation. For fungal differentiation, the amplified rDNA-ITS region was digested with the restriction endonucleases Haelll and Taql and the obtained ITS fragments separated by agarose gel electrophoresis. The specific DNA-fragment fingerprint allowed the identification of an unknown fungal sample by comparison of its pattern with those of control fungi within one day from sampling to diagnosis.
Zusammenfassung
Beschrieben werden genetische Untersuchungen an Hausfäulepilzen, die zu einem Schnelltest zur Diagnose von Hausschwamm, Kellerschwamm und Porenhausschwämmen entwickelt wurden. Die Technik ARDRA-ITS basiert auf dem ‘internal transcribed spacer’ (ITS) der ribosomalen DNS, der mittels der Polymerase-Kettenreaktion vermehrt wurde. Zur Amplifizierung des ITS-Bereiches war Mycel ohne vorherige DNS-Reinigung direkt von Agarkulturen sowie sogar von Pilzmaterial aus einem Gebäude ohne vorherige Reinkultivierung geeignet. Der amplifizierte ITS wurde dann mit den Restriktionsenzymen Haelll und Taql geschnitten und die Fragmente in Agarosegeien elektrophoretisch aufgetrennt. Die spezifischen Fragmentgrößen ermöglichten als “Fingerabdruck“ die Diagnose einer unbekannten Pilzprobe durch Vergleich ihres Musters mit denjenigen von Kontrollpilzen. Der Schnelltest kann innerhalb eines Tages durchgeführt werden.
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Schmidt, O., Moreth, U. Genetic studies on house rot fungi and a rapid diagnosis. Holz als Roh- und Werkstoff 56, 421–425 (1998). https://doi.org/10.1007/s001070050345
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DOI: https://doi.org/10.1007/s001070050345