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Identification of the genus Armillaria by specific amplification of an rDNA‐ITS fragment and evaluation of genetic variation within A. ostoyae by rDNA‐RFLP and RAPD analysis
Author(s) -
Schulze S.,
Bahnweg G.,
Möller E. M.,
Sandermann H.
Publication year - 1997
Publication title -
european journal of forest pathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.535
H-Index - 49
eISSN - 1439-0329
pISSN - 0300-1237
DOI - 10.1111/j.1439-0329.1997.tb00865.x
Subject(s) - biology , restriction fragment length polymorphism , rapd , upgma , genetics , armillaria , intraspecific competition , genetic variation , ribosomal dna , phylogenetic tree , spacer dna , genetic marker , genetic diversity , polymerase chain reaction , botany , population , gene , zoology , demography , sociology
Summary Genetic variation among Armillaria ostoyae isolates was studied by rDNA‐restriction fragment length polymorphism (RFLP) and random amplified polymorphic DNA (RAPD) analysis. A total of 20 A. ostoyae isolates, mainly obtained from Picea spp. of different geographical origins, were examined. Southern hybridization of whole‐cell DNAs digested with Ava II and probed with biotin‐labelled cloned rDNA from Saccharomyces carlsbergensis allowed the differentiation of five RFLP groups. UPGMA cluster analysis of RAPD profiles (138 scorable bands) generated by 10 decamer primers (OPA 01‐OPA 10) grouped the isolates in subclusters at similarity levels between 40% and 96%, indicating high intraspecific genetic variability. Some isolates of different geographical origins subgrouped together, suggesting that similar mutational events have occurred independently and that genetic exchange and recombination occurs among the DNAs in natural populations. The potential role of historical and current spread of spruce plants on the genetic variation of A. ostoyae isolates in Europe is discussed. Using the primer pair ARM‐1 and ARM‐2, an Armillaria‐specific ITS‐DNA fragment of about 660 bp was obtained. No intraspecific RFLP of this amplicon could be revealed, indicating low genetic variability of this region. The established informative RFLP and RAPD markers and also the Armillaria‐specific ITS‐DNA fragment may be powerful tools for further epidemiological, phylogenetic and host‐pathogen interaction studies with A. ostoyae .