Identification and pathogenicity assessment of Colletotrichum isolates causing bitter rot of apple fruit in Belgium
Worldwide Colletotrichum spp. have been identified as a problem in the apple production. This is the first study executed and confirming the presence of Colletotrichum spp. causing the postharvest disease bitter rot on apple fruits in Belgium. The identification, genetic diversity of Colletotrichum isolates (present in Belgian apple orchards) their morphological traits and pathogenicity on two apple cultivars (cvs. Pinova and Nicoter) with a different level of susceptibility were studied. Based on sequence analysis of six different gene regions beta-tubuline (TUB2), histone H3 (HIS3), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), chitin synthase 1 gene (CHS-1), actin (ACT) and the Internal Transcriber Spacer (ITS) gene region, six different Colletotrichum spp., belonging to either the C. acutatum or C. gloeosporioides complexes, were isolated from twenty-one apple cultivars in three Belgian orchards: C. fioriniae, probably C. kahawae, C. salicis, C. rhombiforme, C. acutatum and C. godetiae. Colletotrichum godetiae was found to be the most present and pathogenic species in Belgian orchards. The species C. rhombiforme was found and identified on apple fruit for the first time. Reliable morphological discrimination between species, based on features such as in vitro growth rate, colony colour and spore measurements, is not possible. As such, molecular identification appears to outperform morphological analysis and was in this study the most ideal tool for identifying unknown isolates of Colletotrichum species. Inoculation assays on two apple cultivars revealed a significant difference in pathogenicity among isolates and among Colletotrichum species. The pathogenicity tests also showed that isolates coming from another host species, e.g. strawberry, are also pathogenic on apple fruits. Cultivar Pinova appeared to be more susceptible to bitter rot than cv. Nicoter. Given the difficulties with managing Colletotrichum infections, additional knowledge on the pathogen and the plant-pathogen interaction is essential for effective disease control.
KeywordsPostharvest fungal disease Apple bitter rot Molecular multilocus phylogeny Inoculation assays
The authors thank the Fund for Scientific Research (FWO) Flanders for providing funding for this research (grant number 1S44116N). Thanks to Dalphy Harteveld for her scientific insight concerning this paper.
This study was funded by FWO (research foundation Flanders) Grant number: 1S44116N.
Compliance with ethical statement
Conflict of interest
The authors declare that they have no conflict of interest.
Research involving human participants and/or animals
Not applicable to this study, did not work with humans or animals.
Not applicable to this study, did not work with humans.
- Agrios, G. N. (2005a). Anthracnose diseases caused by ascomycetes and Deureromycetes. In G. N. Agrios (Ed.), Plant pathology (pp. 483–501). Burlington: Elsevier Academic Press.Google Scholar
- Angay, O., Fleischmann, F., Recht, S., Herrmann, S., Matyssek, R., Oßwald, W., Buscot, F., & Grams, T. E. E. (2014). Sweets for the foe – Effects of non-structural carbohydrates on the susceptibility of Quercus robur against Phytophthora quercina. New Phytologist, 203, 1282–1290.CrossRefGoogle Scholar
- Baroncelli, R., Zapparata, A., Sarrocco, S., Sukno, S. A., Lane, C.R., Thon, M.R., Vannacci, G., Holub, E., Sreenivasaprasad, S. (2015). Molecular Diversity of anthracnose pathogen populations associated with UK strawberry production suggests multiple introductions of three different Colletotrichum species. PLoS One Available on: doi: https://doi.org/10.1371/journal.pone.0129140, [13/08/2015].
- Boratyn, G. M., Camacho, C., Cooper, P. S., Coulouris, G., Fong, A., Ma, N., Maden, T. L., Matten, W. T., McGinnis, S. D., Merezhuk, Y., Raytselis, Y., Sayers, E. W., Tao, T., Ye, J., & Zaretskaya, I. (2013). BLAST: A more efficient report with usability improvements. Nucleic Acids Research, 41, 29–33.CrossRefGoogle Scholar
- Børve, J., & Stensvand, A. (2015). Colletotrichum acutatum on apple in Norway. IOBC-WPRS Bulletin, 110, 151–157.Google Scholar
- Cai, L., Hyde, K. D., Taylor, P. W. J., Weir, B. S., Waller, J. M., Abang, M. M., Zhang, J. Z., Yang, Y. L., Phoulivong, S., Liu, Z. Y., Prihastuti, H., Shivas, R. G., McKenzie, E. H. C., & Johnston, P. R. (2009). A polyphasic approach for studying Colletotricum. Fungal Diversity, 39, 183–204.Google Scholar
- Corda, A. C. I. (1831). Die Pilze Deutschlands In: Sturm J (ed.) Deutschlands Flora in Abbildungen nach der Natur mit Beschreibungen, 3. Abtheilung, 12, 1–144.Google Scholar
- Crous, P. W., Groenewald, J. Z., Risede, J. M., & Hywel-Jones, N. L. (2004). Calonectria species and their Cylindrocladium anamorphs: Species with sphaeropedunculate vesicles. Studies in Mycology, 50, 415–430.Google Scholar
- DeLong, J. M., Prange, R. K., & Harrison, P. A. (1999). Using the Streif index as a final harvest window for controlled-atmosphere storage of apples. Horticultural Science, 13, 1251–1255.Google Scholar
- Everett, K. R. (2014). Anthracnose and stem-end rots of tropical and subtropical fruit- new names for old foes. In D. Prusky & M. L. Gullino (Eds.), Post-harvest pathology. Plant pathology in the twenty-first century. Contributions to the 10th international congress, ICPP 2013 (Vol. 7, pp. 55–70). Switzerland: Springer International Publishing.Google Scholar
- Glass, N. L., & Donaldson, G. C. (1995). Development of primer sets designed for the use with the PCR to amplify conserved genes from filamentous ascomycetes. Applied and Environmental Microbiology, 61, 1323–1330.Google Scholar
- Ismail, A. M., Cirvilleri, G., Yaseen, T., Epifani, F., Perrone, G., & Polizzi, G. (2015). Characterisation of Colletotrichum species causing anthracnose disease of mango in Italy. Journal of Plant Pathology, 97, 167–171.Google Scholar
- Leyronas, C., Duffaud, M., & Nicot, P. C. (2012). Compared efficiency of the isolation methods for Botrytis cinerea. Mycology, 3, 221–225.Google Scholar
- Martin-Felix, Y., Groenewald, J. Z., Cai, L., Chen, Q., Marincowitz, S., Barnes, I., Benschn, K., Braun, U., Camporesi, E., Damm, U., de Beer, Z. W., Dissanayake, A., Edwards, J., Giraldo, A., Hernandez-Restrepo, M., Hyde, K. D., Jayawardena, R. S., Lombard, L., & Crous, P. W. (2017). Genera of phytopathogenic fungi: GOPHY 1. Studies in Mycology, 86, 99–216.CrossRefGoogle Scholar
- Nour, V., Trandafir, I., & Ionica, M. A. (2010). Compositional characteristics of fruits of several apple (Malus domestica Borkh.) cultivars. Notulae Botanicae Horti Agrobotanici Cluj-Napoca., 38, 228–233.Google Scholar
- Prihastuti, H., Cai, L., Chen, H., McKenzie, E. H. C., & Hyde, K. D. (2009). Characterization of Colletotrichum species associated with coffee berries in northern Thailand. Fungal Diversity, 39, 89–109.Google Scholar
- SAS Institute Inc. (2013). Using JMP 11. Cary, NC: SAS Institute Inc.Google Scholar
- Sutton, B. C. (1980). The coelomycetes. Fungi imperfecti with pycnidia, acervuli and stromata. CABI, Kew.Google Scholar
- Valero, M., Garcıa-Martınez, S., Giner, M., Alonso, A., & Ruiz, J. (2010). Benomyl sensitivity assays and species-specific PCR reactions highlight association of two Colletotrichum gloeosporioides types and C. acutatum with rumple disease on Primofiori lemons. European Journal of Plant Pathology, 127, 399–405.CrossRefGoogle Scholar
- von Arx, J. A. (1957). Die Arten der Gattung Colletotrichum Corde. Phytopathologische Zeitschrift, 29, 413–468.Google Scholar