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Variations in rDNA ITS of Czech Armillaria species determined by PCR and HPLC

Published online by Cambridge University Press:  01 October 2004

Jan LOCHMAN ,
Omar SERÝ ,
Libor JANKOVSKÝ  and
Vladimir MIKES
Jan LOCHMAN
Affiliation:
Department of Biochemistry, Faculty of Science, Masaryk University, Kotlarska 2, 61137 Brno, Czech Republic. E-mail: mikes@chemi.muni.cz
Omar SERÝ
Affiliation:
Department of Comparative Animal Physiology and General Zoology, Faculty of Science, Masaryk University, Brno, Czech Republic.
Libor JANKOVSKÝ
Affiliation:
Department of Forest Protection and Hunting, Faculty of Forestry, Mendel Agronomy and Forestry University, Zemedelska 1, Brno, Czech Republic.
Vladimir MIKES
Affiliation:
Department of Biochemistry, Faculty of Science, Masaryk University, Kotlarska 2, 61137 Brno, Czech Republic. E-mail: mikes@chemi.muni.cz
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Abstract

We analysed 40 isolates of species Armillaria. borealis, A. cepistipes, A. gallica, A. mellea, A. ostoyae and A. tabescens, mostly collected in the Czech Republic, by PCR-RFLP of the ITS rRNA genes using the restriction endonucleases AluI, HinfI and MboI. Restriction fragments were analysed by ion-exchange high performance liquid chromatography which proved to be more useful informative, and less time-consuming than classical electrophoresis on agarose gel. The HPLC method enabled detection of some heterozygous strains. HinfI discriminated between all six species. Ten isolates were sequenced to confirm changes in restriction sites found by restriction analysis. Cluster analysis based on the restrictions patterns of restriction endonucleases AluI and HinfI divided the analysed species into three groups. The first and the most distant group contained all A. mellea isolates, the second group was formed by A. tabescens and the third group contained species A. borealis, A. cepistipes, A. gallica and A. ostoyae. The A. tabescens group was very homogenous regardless of the origin of isolates (Czech Republic, France and Finland).

Type
Research Article
Information
Mycological Research , Volume 108 , Issue 10 , October 2004 , pp. 1153 - 1161
Copyright
© The British Mycological Society 2004

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