Comparison of mycelial proteomes of two Verticillium albo-atrum pathotypes from hop

Verticillium wilt diseases caused by Verticillium spp. are known in many important crops and can seriously threaten their production. We studied Verticillium albo-atrum by comparative analysis of the proteome of four hop isolates, classified by the severity of wilt symptoms as mild and lethal pathotypes, from two geographic origins. A two-dimensional electrophoresis reference map of mycelium proteins was first established, resolving up to 650 protein spots on Coomassie-stained gels in a range of pH 4-7 and M-W 14 - 116 kDa. The average coefficient of variance for the 268 matched protein spots was 16% and 15%, respectively, for technical and biological variability. Principal component analysis (PCA) discriminated the geographic origin of the isolates and between the two pathotypes and showed a closer relationship among English isolates than Slovene ones. The two-dimensional electrophoresis patterns of one mild (PG1) with one lethal pathotype (PG2) from Slovenia and one mild (M) with one lethal pathotype (PV1) from England were compared. A total of 27 and 30 spots were found differentially expressed between the pathotypes, which were analysed by tandem mass spectrometry. Fifty-three proteins were identified, of which 17 matched proteins with annotated functions. The lethal pathotypes showed increased expression of peroxiredoxine and ascorbate peroxidase, a higher level of cytoskeleton components and regulators, and a higher rate of protein synthesis and energy metabolism. These results reveal differences in the expression level of the identified proteins between the two pathotypes and are discussed in relation to virulence.