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Detection of Heterobasidion annosum s. l. [(Fr.) Bref.] in Norway Spruce by Polymerase Chain Reaction
Authors:G Bahnweg  E M Möller  S Anegg  C Langebartels  O Wienhaus  H Sandermann jr  
Institution:Authors' addresses: GSF –National Research Center for Environment and Health, Institute of Biochemical Plant Pathology, Ingolstädter Landstr. 1, D-85764 Neuherberg, Germany;;University of Hohenheim, Institute of Plant Breeding, Seed Science, and Population Genetics, D-70593 Stuttgart, Germany;;Technical University of Dresden, Institute of Plant Chemistry and Wood Chemistry, D-01737 Tharandt, Germany
Abstract:Internal transcribed spacer (ITS) sequences of the rDNA repeat unit of Heterobasidion annosum were used to design specific primers for the detection and quantification of this important forest pathogen by polymerase chain reaction (PCR). Specificity of detection was cross‐checked against a variety of other fungi (saprophytes, root pathogens, mycorrhizal fungi) which may occur in the same environment. As little as 1 pg fungal DNA (equiv. to 10–40 genomes) could be detected in 200 ng spruce root DNA (from 1 mg fresh spruce root). The Heterobasidion‐specific primers allowed simultaneous detection of Armillaria spp. in multiplex PCR. The method was successfully applied to increment cores of Norway spruce from the forest region Tharandter Wald (Saxonia, Germany), Oberbärenburg (East Ore Mountains, Saxonia) and Oberschleissheim (north of Munich, Bavaria).
Keywords:Heterobasidion                        Armillaria            root and butt rot  rDNA  ITS  specific primers  field application
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