Proteomic analysis of Colletotrichum kahawae-resistant and susceptible coffee fruit pericarps
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Coffee berry disease (CBD) is caused by the fungus Colletotrichum kahawae and is restricted to the African continent, where it generates losses of up to 80 % of coffee production. Weather conditions in certain growing areas at high altitudes in Colombia appear to be very favourable for the development of this disease. Certain genotypes of Coffee arabica are resistant to this pathogen, such as the Timor Hybrid and some Ethiopian accessions. It is important to identify the proteins in these coffee genotypes that are associated with resistance to this fungus. Therefore, we compared the proteomes of two genotypes that are resistant to different isolates of C. kahawae with the proteome of the susceptible coffee genotype Caturra. We optimized the methodology applied for the extraction, cleaning and purification of proteins from the green fruit pericarp at 150 to 170 days after flowering. Through two-dimensional differential gel electrophoresis, proteomic map images were obtained for the resistant and susceptible genotypes. Fifty-two protein spots that were significantly different between the resistant and susceptible genotypes were detected. These protein spots were isolated and sequenced via mass spectrometry. The sequence analysis identified 14 proteins in the Timor Hybrid and 14 in CCC1147 that were associated with resistance and pathogen defence.
KeywordsColletotrichum kahawae CBD Proteome Differential gel electrophoresis Mass spectrometry Coffee berry disease
This research was part of the project “Application of genomic developments for the sustainability of the Colombian coffee crop” under agreement No. 2011–102 between the Ministry of Agriculture and Rural Development of Colombia and the National Federation of Coffee Growers of Colombia (FNC No. 217 of 2011). The authors thank Dr. Ricardo Acuña for collaboration in the development of this research.
- Ernst, K., Kumar, A., Kriseleit, D., Kloos, D. U., Phillips, M. S., & Ganal, M. W. (2002). The broad-spectrum potato cyst nematode resistance gene (Hero) from tomato is the only member of a large gene family of NBS-LRR genes with an unusual amino acid repeat in the LRR region. The Plant Journal, 31(2), 127–136.PubMedCrossRefGoogle Scholar
- Gabriëls, S. H. E. J., Takken, F. L. W., Vossen, J. H., de Jong, C. F., Liu, Q., Turk, S. C. H. J., et al. (2006). cDNA-AFLP combined with functional analysis reveals novel genes involved in the hypersensitive response. Molecular Plant-Microbe Interactions, 19(6), 567–576.PubMedCrossRefGoogle Scholar
- Kvitko, B. H., Park, D. H., Velásquez, A. C., Wei, C. H., Russell, A. B., Martin, G. B., et al. (2009). Deletions in the repertoire of Pseudomonas syringae pv. tomato DC3000 type III secretion effector genes reveal functional overlap among effectors. PloS Pathogens, 5(4), 1–16.CrossRefGoogle Scholar
- Salgado-Guimaraes, B. K. (2007). Proteômica diferencial em clones de Coffea canephora sob condições de déficit hídrico. Minas Gerais: Universidad Federal de Vinosa. Programa de Posgraduación en Fisiología Vegetal. 59 páginas. Trabajo de grado: Magíster Scientae em Fisiología Vegetal.Google Scholar
- Salmeron, J. M., Oldroyd, G. E. D., Rommens, C. M. T., Scofield, S. R., Kim, H. S., Lavelle, D. T., et al. (1996). Tomato Prf is a member of the leucine-rich repeat class of plant disease resistance genes an lies embedded within the Pto kinase gene cluster. Cell, 86(1), 123–133.PubMedCrossRefGoogle Scholar
- Wilkins, M. R., Sanchez, J. C., Gooley, A. A., Appel, R. D., Humphery-Smith, I., Hochstrasser, D. F., et al. (1995). Progress with proteome projects: why all proteins expressed by a genome should be identified and how to do it. Biotechnology and Genetic Engineering Reviews, 13, 19–50.CrossRefGoogle Scholar
- Yang, T., Bar-Peled, L., Gebhart, L., Lee, S. G., & Bar-Peled, M. (2009). Identification of galacturonic acid-1-phosphate kinase, a new member of the GHMP kinase superfamily in plants, and comparison with galactose-1-phosphate kinase. Journal of Biological Chemistry, 284(32), 21526–21535.PubMedCrossRefGoogle Scholar