First Report of Monosporascus cannonballus on Watermelon in Brazil.

In 2008 and 2009, vine decline symptoms were observed in three watermelon (Citrullus lanatus (Thunb.) Matsum. & Nakai) fields located in the municipalities of Mossoró (Rio Grande do Norte State) and Quixeré (Ceará State) in northeastern Brazil. Symptoms included yellowing of crown leaves just prior to harvest and collapse of many of the vines. Mean maximum daily temperatures for the first and second half of the season were 28.6 and 25.1°C, respectively. Affected plants exhibited necrotic root systems and lacked most of the secondary and tertiary feeder roots. Numerous perithecia on the roots contained asci and ascospores characteristics of Monosporascus cannonballus Pollack & Uecker (1,2). Small pieces of primary and secondary roots were surface disinfected and plated onto potato dextrose agar (PDA) medium with 0.5 g liter-1 of streptomycin sulfate and incubated for 7 days at 25°C in the dark. Hyphal tips from all colonies were transferred to PDA and further incubated for 30 to 40 days at 25°C in the dark for subsequent growth and sporulation. Isolations consistently yielded colonies of white mycelium, which became dark grayish after 10 to 15 days, and perithecia with one-spored asci. The internal transcribed spacer regions of ribosomal DNA of isolates 18-5 and 19-1 were sequenced (GenBank Accession Nos. GQ891544 and GQ891545). These sequences were identical to sequences of M. cannonballus (GenBank Accession Nos. AM167936 and AM167937). Pathogenicity of these two isolates was confirmed on watermelon cv. Crimson Sweet in a greenhouse maintained at 25 to 30°C. Inoculum was produced in a sand-oat hulls (Avena sativa) medium (0.5 liter of sand, 46 g of ground oat hulls, and 37.5 ml of distilled water) and incubated at 25°C for 1 month. CFU were quantified by serial dilution using 1% hydroxyethyl cellulose. A sterilized mixture of equal portions (vol/vol) of sand and peat moss was used to fill 17-cm-diameter plastic pots and inoculum was added to produce an inoculum concentration of 20 CFU g-1. Five watermelon seeds planted in each pot were later thinned to one seedling per pot. There were five replicated pots for each treatment with an equal number of noninfested pots. Plants were evaluated for disease 45 days after sowing. All isolates of M. cannonballus were highly aggressive and caused severe root necrosis compared with the noninoculated controls. M. cannonballus was reisolated from symptomatic plants, confirming Koch's postulates. In 2004, M. cannonballus was reported in the same Brazilian cucurbit-growing areas causing root rot and vine decline of muskmelon (Cucumis melo L.) (3), but to our knowledge, this is the first report of M. cannonballus on watermelon in Brazil. References: (1) R. D. Martyn and M. E. Miller. Plant Dis. 80:716, 1996. (2) F. G. Pollack and F. A. Uecker. Mycologia 66:346, 1974. (3) R. Sales Jr. et al. Plant Dis. 88:84, 2004.