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 MW 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.
KeywordsFungi Proteomic Two-dimensional electrophoresis Virulence
This study was funded by the Ministry of Higher Education, Science and Technology; contracts no. L4-7179, P4-0077 and S4-486-116/1000-05-310050. The authors thank Dr. Rajcevic Uros from NorLux Neuro-Oncology/Crp Sante, Luxembourg, for help in interpreting MS/MS data. We are grateful to Dr. Jesus Jorrin from the Department of Biochemistry and Molecular Biology, University of Cordoba, Cordoba, Spain, for critical reading of the manuscript and for thoughtful suggestions.
- Aisif, A. R., Oellerich, M., Amstrong, V. W., Riemenschneider, B., Monod, M., & Reichard, U. (2006). Proteome of conidial surface associated proteins of Aspergillus fumigatus reflecting potential vaccine candidates and allergens. Journal of Proteome Research, 5, 954–962. doi: 10.1021/pr0504586.CrossRefGoogle Scholar
- Beckman, C. H. (1987). The nature of wilt disease in plants. St Paul, MN: The American Phytopathological Society.Google Scholar
- Engelhard, A. W. (1957) Host index of Verticillium albo-atrum Reinke and Berth, (including Verticillium dahliae Kleb.). In: Supplement to Plant Disease Reporter No. 244, pp. 23-49.Google Scholar
- Fernandez-Acero, F. J., Jorge, I., Calvo, E., Vallejo, I., Carbu, M., Camafeita, E., et al. (2007). Proteomic analysis of phytopathogenic fungus Botrytis cinerea as a potential tool for identifying pathogenicity factors, therapeutic targets and for basic research. Archives of Microbiology, 187, 207–215. doi: 10.1007/s00203-006-0188-3.PubMedCrossRefGoogle Scholar
- Harris, R. V. (1927). A wilt disease of hops. In: East Malling Research Station Annual Report for 1925, Supplement II. pp. 92–93.Google Scholar
- Jamnik, P., Radisek, S., Javornik, B., & Raspor, P. (2006). 2-D Separation of Verticillium albo-atrum proteins. Acta Agriculturae Slovenica, 87, 455–460.Google Scholar
- Jorge, I., Navarro, R. M., Lenz, D., Ariza, D., Porras, C., & Jorrin, J. (2005). The Holm Oak leaf proteome: Analytical and biological variability in the protein expression level assessed by 2-DE and protein identification tandem mass spectrometry de novo sequencing and sequence similarity search. Proteomics, 5, 222–234. doi: 10.1002/pmic.200400893.PubMedCrossRefGoogle Scholar
- Sewell, G. W. F., & Wilson, J. F. (1974). Hop wilt, soil temperature and nitrogen. In: East Malling Research Station Annual Report for1973. pp. 203–204.Google Scholar
- Ueda, T., Kikuchi, A., Ohga, N., Yamamoto, J., & Takai, Y. (1990). Purification and characterization from bovine brain cytosol of a novel regulatory protein inhibiting the dissociation of GDP from and the subsequent binding of GTP to RhoB p20, a RAS p21-like GTP-binding protein. Journal of Biological Chemistry, 265, 9373–9380.PubMedGoogle Scholar
- Wilhelm, S. (1955). Longevity of the Verticillium wilt fungus in the laboratory and in the field. Phytopathology, 45, 180–181.Google Scholar