A Severe Outbreak of Charcoal Rot in Cantaloupe Melon Caused by Macrophomina phaseolina in Chile

A severe outbreak of charcoal rot was observed in cantaloupe melon (Cucumis melo L.) in the summer of 2011 to 2012 in Curacaví Valley, Chile. Prior to harvest, of 72 plants per cultivar, charcoal rot prevalence varied from 32% to 82% in cvs. Colima, Charantias, Navigator, Origami, Otero, and Samoa. Symptoms were wilting and leaf browning associated with water-soaked lesions at the base of the crown with amber to dark brown exudates. Lesions dried out progressively, turned tan, and cracked. Affected plants declined and died before harvest. Reddish fruit decay was observed. Symptomatic stem and root samples (n = 97) were collected, surface disinfected (96% ethanol, 30 s), plated on PDA acidified with 0.5 ml/liter of 92% lactic acid (APDA), and incubated at 20 ± 1°C. A white, fast-growing mycelium was obtained that turned gray to black after 7 days due to the presence of spherical to oblong black microsclerotia 136 ± 52 μm (n = 80) in diameter. On the basis of colony morphology and microsclerotia, 57 isolates (59%), obtained from 97 melon samples, were tentatively identified as Macrophomina phaseolina (Tassi) Goid. (2,3). The morphological identification of four isolates M1HB-B, M2CO-B, M3CH-R, and M4OT-B (GenBank Accession Nos. JX203630, JX203631, JX203632, and JX203633) was confirmed by sequencing of the internal transcribed spacer region (ITS1-5.8S-ITS2) of rDNA, using primers ITS4 and ITS5, with >99% similarity with the sequences of M. phaseolina (GenBank Accession No. HQ660592) (4). Pathogenicity tests were conducted with isolates M1HB-B, M2CO-B, M3CH-R, and M4OT-B on melon fruits cvs. Colima, Origami, Charantias, and Diva. Four mature melon fruits per cultivar per isolate were surface disinfected with 0.5% sodium hypochlorite for 2 min before inserting a mycelium plug (19 mm²) in a 6 mm diameter hole made with a sterile cork borer. An equal number of perforated fruits in which a sterile agar plug was inserted were left as non-inoculated controls. After 8 days of incubation at 20°C, inoculated fruits developed a spherical, reddish, soft necrotic lesion of 15 to 20 mm in diameter in the pulp. Non-inoculated fruits remained symptomless. The pathogenicity of the four isolates was also studied in 3-month-old melon plants (n = 4) cvs. Colima and Navigator. Plants were inoculated by inserting a mycelial plug (9 mm²) underneath the epidermis of the crown, 5 cm above the soil level. The inoculation site was immediately wrapped with Parafilm to avoid dehydration. An equal number of non-inoculated, but injured plants, treated with a sterile agar plug, were left as controls. After 21 days of incubation under greenhouse conditions (17 ± 5.5°C), all inoculated plants developed water-soaked to dry necrotic lesions, 20 to 70 mm long, yellow to tan in color. No symptoms were obtained in non-inoculated controls. M. phaseolina was reisolated in 84% and 100% of the inoculated plants and fruits, respectively. To our knowledge, this study is the first report of charcoal rot in cantaloupe melon in Chile, previously found on watermelon and melon group inodorus (1). Charcoal rot appears as an emerging disease that aggressively affects current cantaloupe melon cultivars in central Chile. References: (1) G. Apablaza. Cien. Inv. Agr. 20:101, 1993. (2) B. D. Bruton and E. V. Wann. Charcoal rot. Page 9 in: Compendium of Cucurbit Diseases. T. A. Zitter, D. L. Hopkins, and C. E. Thomas, eds. APS, St. Paul, MN, 1996. (3) S. Kaur et al. Crit. Rev. Microbiol. 38:136, 2012. (4) J. Q. Zhang et al. Plant Dis. 95:872, 2011.