Variations in rDNA ITS of Czech Armillaria species determined by PCR and HPLC.
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).
Lochman
Jan
J
Department of Biochemistry, Faculty of Science, Masaryk University, Kotlarska 2, 61137 Brno, Czech Republic.
Serý
Omar
O
Jankovský
Libor
L
Mikes
Vladimir
V
D000363:Agaricales / Q000235:genetics*; D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D018153:Czech Republic; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics*; D021903:DNA, Ribosomal Spacer / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D025282:Mycelium / Q000235:genetics; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D012335:RNA, Ribosomal / Q000737:chemistry / Q000235:genetics; D016415:Sequence Alignment
D004271:DNA, Fungal; D021903:DNA, Ribosomal Spacer; D012335:RNA, Ribosomal