First Report of Gummy Stem Blight Caused by Didymella bryoniae on Watermelon and Confirmation of the Disease on Pumpkin in Tanzania

Abstract

Foliar, stem, and fruit lesions were observed on watermelon (Citrullus lanatus (Thunb.) Matsum. & Nakai) and pumpkin (Cucurbita maxima Duchesne) in two separate research fields in the district of Arusha, Tanzania during the warm, rainy season from February to April 2010. Similar symptoms were observed in commercial watermelon fields and intercropped pumpkin fields in Same and Moshi districts with as much as 100% fruit loss in watermelon. Disease symptoms on watermelon were dark brown, V-shaped leaf lesions. On pumpkin, V-shaped leaf lesions were light brown. On both hosts, stems showed water-soaked lesions after rain, which dried up and cracked. On pumpkin, a gummy, amber exudate was seen after rain on stem and fruit lesions. Flowers and fruits of both hosts developed black rot spots and aborted. Isolation of the causal agent on potato dextrose agar (PDA) from leaf and stem pieces of watermelon and pumpkin plants in Arusha showed white-to-olivaceous green mycelium. Pycnidia formed on one-quarter-strength PDA and produced hyaline, oblong conidia mainly with two guttules, nonseptate, 5 to 11 × 3 to 5 μm. Pathogenicity was tested with three isolates from watermelon and one from pumpkin on four 1-month-old plants per watermelon cvs. Sugar Baby and Charleston Grey and pumpkin cv. Small Sugar per isolate. The test was repeated on the watermelon cultivars. One site on the main stem and two leaves per plant were misted, pricked with a scalpel, inoculated with 3-day-old mycelial plugs (5 × 5 mm), and kept humid at 20 to 30°C in cellophane bags for 3 days. All plants developed leaf and/or stem lesions. Detached, misted leaves were also laid on 2% water agar and inoculated as above. Water-soaked lesions developed around inoculation sites and microscopy of infected tissue revealed pycnidia with conidia as described above. All isolates infected both hosts. A set of control plants and detached leaves, mock inoculated with agar plugs, remained symptomless. The fungus was reisolated from infected leaves and stems of both hosts. On the basis of the morphological characteristics, the fungus was identified as Didymella bryoniae (Auersw.) Rehm (anamorph Phoma cucurbitacearum (Fr.:Fr.) Sacc.) (1,3) and this was confirmed by amplification of species-specific PCR products. The isolates from both hosts were cultured in liquid medium, and DNA was extracted using a DNeasy Plant Mini Kit (Qiagen, Valencia, CA). PCR and multiplex PCR involving D. bryoniae-unique primer sequences D6 and D7S, in combination with primer UNLO28S22, produced the expected band sizes (2). To our knowledge, this is the first report of gummy stem blight and black fruit rot of watermelon caused by D. bryoniae in Tanzania, which confirms a previous report of leaf spot on pumpkin (4), and the first report of black fruit rot on pumpkin. The disease was previously an unidentified problem in watermelon and the severe outbreak was associated with favorable weather conditions. References: (1) A. P. Keinath et al. Phytopathology 85:364, 1995. (2) C. A. Koch and R. S. Utkhede. Can. J. Plant Pathol. 26:291, 2004. (3) E. Punithalingam and P. Holliday. No. 332 in: Descriptions of Pathogenic Fungi and Bacteria. CMI, Kew, Surrey, UK, 1972. (4) E. A. Riley. Mycol. Pap. 75:1, 1960.