Morphology, Molecular Identity, and Pathogenicity of Verticillium dahliae and V. longisporum Associated with Internally Discolored Horseradish Roots.

Abstract

During 2008 to 2009, 255 isolates of Verticillium were obtained from internally discolored horseradish roots collected from California, Illinois, and Ontario. Twenty representative isolates were selected according to morphological features and geographic origin for further characterization. Based on the conidial size, the isolates were divided into two groups: Verticillium dahliae (4.4 ± 1.23 μm) and V. longisporum (7.8 ± 1.76 μm). Genetic diversity of the isolates was determined by sequence analysis of the internal transcribed spacer (ITS) and two mitochondrial genes (cytochrome oxidase subunit III [cox3] and NADH dehydrogenase subunit I [nad1]). Based on ITS analysis, Verticillium isolates were divided into two clades: V. dahliae and V. longisporum. However five isolates of V. longisporum (identified based on conidial size) were clustered with a V. dahliae clade, whereas the other five isolates formed a distinct V. longisporum clade. Combined analysis of the mitochondrial genes cox3 and nad1 showed that the two genetic clades of V. longisporum in ITS region analysis corresponded to the previously reported V. longisporum lineage A1/D3 and A1/D2. Pathogenicity tests revealed that all tested Verticillium isolates caused internal discoloration of horseradish roots, and there were no significant differences in either incidence or severity of root discoloration among the genetic groups.