Discrimination of some African Armillaria species by isozyme electrophoretic analysis

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Abstract
Ten Armillaria isolates, collected from various host plants and widespread geographical origins in tropical Africa, were cultivated on orange fragments in the presence of water and ran different culture media in order to optimize enzyme and mycelial cord production. Seven enzymes involved in the primary metabolism of nitrogen and carbon (glutamate dehydrogenases, aspartate aminotransferase, malate dehydrogenase, glucose n-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase and isocitrate dehydrogenase) were extracted from the mycelial cords and analyzed by polyacry lamide gel electrophoresis. Cluster analysis based on calculated similarity values derived from isozyme banding patterns separated the isolates into five groups. Two isolates considered as belonging to A. mellea ssp. africana (an African species closely related to European A. mellea) were present in a clearly Separated cluster when compared to the other isolate groups. Two Kenyan isolates, belonging to an as yet unnamed biological species, which were characterized by the production of few slow growing mycelial cords, were also found in a separate cluster with slightly greater similarity coefficients to the other isolates. The six other isolates, referred to as isolates of A. heimii (a highly variable species with different sexual systems) fell into three sub-clusters of variable homology. The two homothallic heimii isolates from Tanzania and Malawi, which were very closely related, displaying 100% isozyme similarity, exhibited a higher degree of similarity with the two other homothallic heimii isolates from Zimbabwe and Congo, than with the two heterothallic unifactorial heimii isolates from Cameroon and Gabon. The value of isozymes in the classification of African Armillaria spp. is discussed.