HomePlant DiseaseVol. 103, No. 7First Report of Anthracnose Causing Both Crown and Fruit Rot of Strawberry by Colletotrichum siamense in North Carolina PreviousNext DISEASE NOTESFirst Report of Anthracnose Causing Both Crown and Fruit Rot of Strawberry by Colletotrichum siamense in North CarolinaT. B. Adhikari, J. G. Chacon, G. E. Fernandez, and F. J. LouwsT. B. Adhikari†Corresponding author: T. B. Adhikari; E-mail Address: [email protected]http://orcid.org/0000-0001-7118-6875Department of Entomology and Plant Pathology, North Carolina State University, Raleigh, NC 27695Search for more papers by this author, J. G. ChaconDepartment of Horticultural Science, North Carolina State University, Raleigh, NC 27695Search for more papers by this author, G. E. FernandezDepartment of Horticultural Science, North Carolina State University, Raleigh, NC 27695Search for more papers by this author, and F. J. LouwsDepartment of Entomology and Plant Pathology, North Carolina State University, Raleigh, NC 27695Department of Horticultural Science, North Carolina State University, Raleigh, NC 27695Search for more papers by this authorAffiliationsAuthors and Affiliations T. B. Adhikari1 † J. G. Chacon2 G. E. Fernandez2 F. J. Louws1 2 1Department of Entomology and Plant Pathology, North Carolina State University, Raleigh, NC 27695 2Department of Horticultural Science, North Carolina State University, Raleigh, NC 27695 Published Online:10 May 2019https://doi.org/10.1094/PDIS-02-19-0314-PDNAboutSectionsSupplemental ToolsAdd to favoritesDownload CitationsTrack Citations ShareShare onFacebookTwitterLinked InRedditEmailWechat Anthracnose fruit rot and anthracnose crown rot of strawberries (Fragaria × ananassa Duch.), caused by Colletotrichum spp., have been mainly associated with the C. acutatum complex (Baroncelli et al. 2015; Damm et al. 2012) and the C. gloeosporioides complex (Weir et al. 2012), respectively. In September 2017, typical symptoms of anthracnose were observed on strawberry plants in a propagation greenhouse in North Carolina, U.S.A. Symptoms that appeared were crown rotting, wilting, and stunting. Small pieces (2 mm2) of necrotic tissue were cut with a sterile scalpel from the crown lesion. Approximately 15% of the plants showed such typical symptoms. The diseased segment was surface sterilized with 70% ethanol for 30 s and placed on acidified potato dextrose agar (APDA). Hyphal tips from colonies emerging from the edge of the tissue were transferred onto fresh APDA Petri plates and incubated at 25°C. Morphological characteristics after 7 days of incubation showed light-gray to whitish aerial mycelium. Conidia were hyaline, aseptate, fusiform with obtuse ends, cylindrical, 13.2 to 17.5 µm long, and 4.9 to 7.5 µm wide. These characteristics of conidia were matched with some species belonging to the C. gloeosporioides complex including C. siamense (Weir et al. 2012). For accurate identification, genomic DNA of isolate 28244 was extracted and amplified with partial sequences of actin (ACT), chitin synthase (CHS), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), ribosomal internal transcribed spacer (ITS), and manganese-superoxide dismutase (SOD2) primer sets (Weir et al. 2012). Sequencing of the purified polymerase chain reaction products was performed at the Genomic Sciences Laboratory, North Carolina State University, Raleigh, NC (White et al. 1990). Sequences of the amplified regions of isolate 28244 were deposited to GenBank (accession nos. MH722310 to MH722314). Additionally, sequences of 22 Colletotrichum species within the C. gloeosporioides complex (Weir et al. 2012) were retrieved from GenBank. Jukes–Cantor neighbor-joining trees inferred from the concatenated sequences (ACT, CHS, GAPDH, ITS, and SOD2) placed isolate 28244 from North Carolina within the same cluster of C. siamense including isolate BRSP09 from Bangladesh (Gupta et al. 2018). These sequences were compared with the GenBank database using BLASTn and revealed a high percentage of similarity (>99%). To confirm pathogenicity of the fungus, Koch’s postulates were performed by three methods. Three-month-old strawberry plants of cultivar ‘Camarosa’ were either spray inoculated on leaves with a conidial suspension (106 conidia/ml) or injected (10 μl) into the crown of each plant. Ten plants inoculated with distilled water served as controls. Plants were covered with plastic bags for 48 h and placed in the greenhouse at 28°C. Leaf and crown rot symptoms were monitored weekly after inoculation. No symptoms or spores developed on leaves, which was also confirmed by a paraquat assay after the first 3 weeks. However, dark brown necrotic lesions were observed on the crowns 4 weeks after inoculation. The fungus was reisolated from the inoculated crown tissues. In another experiment, 10 immature (half-green and half-yellow) strawberry fruit (cv. ‘Chandler’) were surface sterilized with 70% ethanol and then 0.5% sodium hypochlorite for 30 s and rinsed with sterile distilled water three times. Fruit was injected with 10 μl of conidial suspension (106 conidia/ml) and placed in a plastic crisper container. Sterile distilled water–inoculated fruits served as controls. Fruits in the containers were incubated at 25°C in the dark. All experiments were conducted twice. Fruit rot symptoms were observed on fruits 3 days after inoculation, and the control fruits remained asymptomatic. Within the C. gloeosporioides species complex, C. aenigma, C. fructicola, C. siamense, and C. theobromicola were previously reported on strawberries (Baroncelli et al. 2015; Weir et al. 2012). More recently, C. siamense has been reported to cause crown rot of strawberry in Bangladesh (Gupta et al. 2018) and fruit rot on strawberry in Brazil (Capobiango et al. 2016). To our best knowledge, this is the first report of anthracnose causing both crown and fruit rot of strawberry by C. siamense, one of the species within the C. gloeosporioides complex, in North Carolina. This disease can seriously affect strawberry fruit quality and yield and is capable of producing resistance to Fungicide Resistance Action Committee code 11 fungicides (Hu et al. 2015); thus, effective disease management strategies should be investigated and implemented.The author(s) declare no conflict of interest.References:Baroncelli, R. et al. 2015. PLoS One 10:e0129140. https://doi.org/10.1371/journal.pone.0129140 Crossref, ISI, Google ScholarCapobiango, N. P. et al. 2016. Plant Dis. 100:859. https://doi.org/10.1094/PDIS-10-15-1121-PDN Link, ISI, Google ScholarDamm, U. et al. 2012. Stud. Mycol. 73:37. https://doi.org/10.3114/sim0010 Crossref, ISI, Google ScholarGupta, D. R. et al. 2018. Plant Dis. 103:580. https://doi.org/10.1094/PDIS-08-18-1461-PDN Link, ISI, Google ScholarHu, et al. 2015. Plant Dis. 99:806. https://doi.org/10.1094/PDIS-10-14-1077-RE Link, ISI, Google ScholarWeir, B. S., et al. 2012. Stud. Mycol. 73:115. https://doi.org/10.3114/sim0011 Crossref, ISI, Google ScholarWhite, T. J., et al. 1990. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA. Crossref, Google ScholarThe author(s) declare no conflict of interest.DetailsFiguresLiterature CitedRelated Vol. 103, No. 7 July 2019SubscribeISSN:0191-2917e-ISSN:1943-7692 DownloadCaptionApple cultivar Joya Cripps Red lesions caused by Colletotrichum fructicola (Nodet et al.). Photo credit: P. Nodet. Symptoms of Lotus powdery mildew caused by Erysiphe takamatsui (Zhou et al.). Photo credit: C. Liang. 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