Molecular differentiation and detection of ginseng-adapted isolates of the root rot fungus Cylindrocarpon destructans

The soilborne fungus Cylindrocarpon destructans (teleomorph: Neonectria radicicola) causes root rot in a wide range of plant hosts; the disease is of particular concern in ginseng production, and in conifer and fruit tree nurseries. beta-Tubulin gene and rRNA gene internal transcribed spacer (ITS) sequence data and pathogenicity assays were used to characterize isolates of C. destructans from ginseng and other hosts. The results of these studies demonstrated a high amount of sequence divergence among strains identified as C. destructans or N. radicicola, suggesting the existence of several phylogenetic species in this complex. Accordingly. we propose that the two varieties of N. radicicola be raised to species status. Certain highly aggressive ginseng isolates from Ontario, Korea, and Japan have identical ITS and beta-tubulin sequences, and form a monophyletic clade (designated "clade a"); these strains are identified as C. destructans f. sp. panacis. Other ginseng strains clustered in monophyletic groups with strains from angiosperm and conifers. A subtractive hybridization method was used to isolate genomic DNA sequences with diagnostic potential from the aggressive C. destructans Ontario ginseng isolate 1640. One of these sequences was similar to the rRNA gene intergenic spacer from a Fusarium oxysporum isolate from Pinus ponderosa, and hybridized to DNA from E oxysporum and all C. destructans isolates tested. Primers were designed that could be used to amplify this sequence specifically from the highly aggressive, ginseng-adapted C. destructans isolates from Ontario and Korea and other members of clade a.