Abstract
Anthracnose caused by Colletotrichum spp., is one of the most serious diseases affecting tamarillo (Solanum betaceum) production in the Ecuadorian highlands. The objective of this study was to characterise Colletotrichum isolates obtained from tamarillo to clarify its taxonomic and phylogenetic position. Based on phenotypic and morphologic characterisation, the isolates of this study were consistently grouped within the Colletotrichum acutatum complex. Multilocus molecular phylogenetic analysis based on combined sequences of actin (ACT), β-tubulin (TUB2), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) sequences using Bayesian probabilities, indicated that 16 of 20 isolates from Imbabura, Cotopaxi, Tungurahua, Bolivar, Chimborazo, Azuay and Loja provinces, belonged to C. tamarilloi. This study represents the first reported case of anthracnose of S. betaceum caused by C. tamarilloi in the Ecuadorian highlands. Interestingly, pathogenicity tests and multilocus molecular analysis revealed a new infraspecific and more aggressive species formed by four isolates obtained from the Pichincha province. These findings probably provided interesting information related to mutant isolates and a possible description of a new pathogenic species affecting tamarillo fruits.
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Afanador-Kafuri, L., Minz, D., Maymon, M., & Freeman, S. (2003). Characterization of Colletotrichum isolates from tamarillo, passiflora, and mango in Colombia and identification of a unique species from the genus. Phytopathology, 93, 579–587.
Bailey, J. A., O’Connell, R. J., Pring, R. J., & Nash, C. (1992). Infection strategies of Colletotrichum species. In In: Colletotrichum: biology, pathology and control. Wallingford, Gran Bretaña: CAB International.
Blank, R. H., Dance, H. M., Hampton, R. E., Olson, M. H., & Holland, P. T. (1987). Tamarillo (Cyphomandra betacea): effect of field-applied fungicides and post-harvest fungicide dips on storage rots of fruit. New Zealand Journal of Experimental Agriculture, 15, 191–198.
Brown, A. E., Sreenivasaprasad, S., & Timmer, L. W. (1996). Molecular characterization of slow-growing orange and key lime anthracnose strains of Colletotrichum from Citrus as C. acutatum. Phytopathology, 86, 523–527.
Buddie, A., Martínez-Culebras, P., Bridge, P., García, M., Querol, A., Cannon, P., & Monte, E. (1999). Molecular characterization of Colletotrichum strains derived from strawberry. Mycological Research, 103, 385–394.
Carbone, I., & Kohn, L. M. (1999). A method for designing primer sets for speciation studies in filamentous ascomycetes. Mycologia, 91, 553–556.
Castellanos, C., & Mosquera, G. (2011). Producción de micelio en medio líquido para extracción de ADN. Guías Prácticas de Laboratorio para el Manejo de Patógenos del Frijol. Costa Rica: CIAT.
Centro de Información e Inteligencia Comercial (CICO-CORPEI). (2009). Perfiles de producto, perfil de tomate de árbol. Ecuador-Ibarra, pp. 23.
Choi, Y. W., Hyde, K. D., & Ho, W. H. (1999). Single spore isolation of fungi. Fungal Diversity, 3, 29–38.
Chull, Y., Jung, Y., Woo, K., Jung, S., & Woo, S. (2008). Ultrastructures of Colletotrichum orbiculare in the leaves of cucumber plants expressing induced systemic resistance mediated by Glomus intraradices BEG110. Mycobiology, 36, 236–241.
Chung, W. H., Ishii, H., Nishimura, K., Fukaya, M., Yano, K., & Kajitani, Y. (2006). Fungicide sensitivity and phylogenetic relationship of anthracnose fungi isolated from various fruit crops in Japan. Plant Disease, 90, 506–512.
Crouch, J. A., Clarke, B. B., & Hillman, B. I. (2009). What is the value of ITS sequence data in Colletotrichum systematics and species diagnosis? A case study using the falcate-spored graminicolous Colletotrichum group. Mycologia, 101, 648–656.
Damm, U., Cannon, P. F., Woudenberg, J., & Crous, P. W. (2012). The Colletotrichum acutatum species complex. Studies in Mycology, 73, 37–113.
Du, M., Schardl, C. L., Nuckles, E. M., & Vaillancourt, L. J. (2005). Using mating-type gene sequences for improved phylogenetic resolution of Colletotrichum species complexes. Mycologia, 3, 641–658.
Falconí, C. E., Visser, R. G., & Van Heusden, A. W. (2013). Phenotypic, molecular, and pathological characterization of Colletotrichum acutatum associated with Andean lupine and tamarillo in the Ecuadorian Andes. Plant Disease, 97, 819–827.
Freeman, S., & Rodríguez, R. J. (1995). Differentiation of Colletotrichum species responsible for anthracnose of strawberry by arbitrarily primed PCR. Mycological Research, 99, 501–504.
Freeman, S., Katan, T., & Shabi, E. (1998). Characterization of Colletotrichum species responsible for anthracnose diseases of various fruits. Plant Disease, 82, 596–605.
Guerber, J., Liu, B., Correll, J., & Johnston, P. (2003). Characterization of diversity in Colletotrichum acutatum sensu lato by sequence analysis of two gene introns, mtDNA and intron RFLPs, and mating compatibility. Mycologia, 95, 872–895.
Gunnell, P. S., & Gubler, W. D. (1992). Taxonomy and morphology of Colletotrichum species pathogenic to strawberry. Mycologia, 84, 157–165.
Kearse, M., Moir, R., Wilson, A., Stones-Havas, S., Cheung, M., Sturrock, S., Buxton, S., Cooper, A., Markowitz, S., Duran, C., Thierer, T., Ashton, B., Mentjies, P., & Drummond, A. (2012). Geneious basic: an integrated and extendable desktop software platform for the organization and analysis of sequence data. Bioinformatics, 28, 1647–1649.
Kongtragoul, P., Nalumpang, S., Miyamoto, Y., Izumi, Y., & Akimitsu, K. (2011). Mutation at codon 198 of Tub2 gene for Carbendazim resistance in Colletotrichum gloeosporioides causing mango anthracnose in Thailand. Journal of Plant Protection Research, 57, 377–383.
León, J., Viteri, P., & Cevallos, G. (2004). Manual del Cultivo de Tomate de Árbol (Solanum betaceum Cav.). Ecuador-Quito, INIAP. Manual No. 61.
Michaelsen, A., Pinzari, F., Barbabietola, N., & Piñar, G. (2013). Monitoring the effects of different conservation treatments on paper-infecting fungi. International Biodeterioration & Biodegradation, 84, 333–341.
Nirenberg, H. I., Feiler, U., & Hagedorn, G. (2002). Description of Colletotrichum lupine comb.nov.in modern terms. Mycologia, 94, 307–320.
O’Donnell, K., & Cigelnik, E. (1997). Two divergent intragenomic rDNA ITS2 types within a monophyletic lineage of the fungus Fusarium are nonorthologous. Molecular Phylogenetics and Evolution, 7, 103–116.
Photita, W., Taylor, P. W., Ford, R., Hyde, K. D., & Lumyong, S. (2005). Morphological and molecular characterization of Colletotrichum species from herbaceous plants in Thailand. Fungal Diversity, 18, 117–133.
Prohens, J., & Nuez, F. (2001). The Tamarillo (Cyphomandra betacea): a review of a promising small fruit crop. Small Fruits Review, 1, 43–68.
Rambaut, A., & Drummond, A. (2010). TreeAnnotator version 1.6.1 [computer program] http://beast.bio.ed.ac.uk.
Ronquist, F., & Huelsenbeck, J. (2003). MrBayes 3: Bayesian phylogenetic inference under mixed models. Bioinformatics, 19, 1572–1574.
Santillán, F. (2001). Manual del cultivo sustentable de tomate de árbol (p. 18). Ecuador: Universidad de Cuenca.
Santos, S., Tozze, H., Corrêa, D., Quintão, G., & Massola, N. (2013). Etiology and pathogenicity of two different isolates of Colletotrichum spp. obtained from physic nut seeds. Journal of Seed Science, 35, 139–146.
Seifert, K. A. (2009). Progress towards DNA barcoding of fungi. Molecular Ecology Resources, 9, 83–89.
Shagai-Maroof, M., Soliman, K., Jorgensen, R., & Allard, R. (1984). Ribosomal DNA spacer-length polymorphism in barley: mendelian inheritance, chromosomal locations and population dynamics. Proceedings of the National Academy of Sciences, 81, 8014–8018.
Simmonds, J. H. (1965). A study of the species of Colletotrichum causing ripe fruit rots in Queensland. Queensland Journal of Agricultural and Animal Science, 22, 437–459.
Sreenivasaprasad, S., & Talhinhas, P. (2005). Genotypic and phenotypic diversity in Colletotrichum acutatum, a cosmopolitan pathogen causing anthracnose on a wide range of hosts. Molecular Plant Pathology, 6, 361–378.
Tamura, K., Dudley, J., Nei, M., & Kumar, S. (2013). MEGA6.0: molecular evolutionary genetics analysis (MEGA) software version 6.06. Molecular Biology and Evolution, 30, 2725–2729.
Than, P. P., Shivas, R. G., Jeewon, R., Pongsupasamit, S., Marney, T. S., Taylor, P. W., & Hyde, K. D. (2008). Epitypification and phylogeny of Colletotrichum acutatum J. H. Simmonds. Fungal Diversity, 28, 97–108.
Weir, B., Johnston, P., & Damm, U. (2012). The Colletotrichum gloeosporioides species complex. Studies in Mycology, 73, 115–180.
Yarden, O., & Katan, T. (1993). Mutations leading to substitutions at amino acid 198 and 200 of beta-tubulin that correlate with benomyl resistant phenotypes of field strains of Botrytis cinerea. Phytopathology, 83, 1478–1483.
Acknowledgments
This research received collaboration of the following institutions: AGROCALIDAD-Tumbaco and IZC (International Zoonosis Center) situated in the Central University of Ecuador. We thank Dr. Lydia Rivera for her valuable contributions (University of Puerto Rico-Mayagüez campus) reviewing this manuscript.
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Caicedo, J.D., Lalangui, K.P., Pozo, A.N. et al. Multilocus molecular identification and phylogenetic analysis of Colletotrichum tamarilloi as the causal agent of Tamarillo (Solanum betaceum) anthracnose in the Ecuadorian highlands. Eur J Plant Pathol 148, 983–996 (2017). https://doi.org/10.1007/s10658-017-1155-3
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DOI: https://doi.org/10.1007/s10658-017-1155-3