First Report of Gummy Stem Blight Caused by Didymella bryoniae on Grafted Watermelon in Tunisia

During the summer of 2006, severe losses were observed in grafted watermelons in the Testour Region in northern Tunisia. Disease symptoms included stem cankers and necrosis and rot of the grafting area that extended a few centimeters along watermelon vines with the production of a brown gummy exudation. Lesions were not observed on leaves or nongrafted plants. Affected plants wilted and eventually died. The presence of small pseudothecia as black specks was observed embedded in the cankers. Isolations from the stems and crown of symptomatic plants onto potato dextrose agar (PDA) supplemented with 0.5 mg/ml of streptomycin sulfate consistently yielded cultures of a fungal agent. These isolates were transferred to PDA and V8 juice agar and incubated at 23°C for 1 month with a 12-h photoperiod. On PDA, they produced numerous pycnidia with hyaline, cylindrical, one-septate conidia, with mean dimensions of 6.7 × 2.5 μm. On V8 juice agar, they produced sparse ostiolate pseudothecia with bitunicate asci and hyaline, oval, one-septate ascospores, with mean dimensions of 13.7 × 5.1 μm. On the basis of these characters, the isolates were identified as Didymella bryoniae (anamorph Phoma cucurbitacearum) (1,2). To further confirm this identification, the complete internal transcribed spacer regions 1 and 2, including the 5.8S ribosomal DNA, of isolates Di-3 and Di-4 were sequenced (GenBank Accession Nos. EF107641 and EF 107642). These sequences were identical to sequences in GenBank from isolates of D. bryoniae (Accession Nos. AF297228 and AF495850). Pathogenicity tests were conducted on watermelon seedlings cv. Giza and Cucurbita hybrid rootstock seedlings cv. Strong Toza using two isolates, Di-3 and Di-4. Seedlings were inoculated at the two- to three-leaf stage. A 5-mm diameter agar disc, cut from the margin of an 8-day-old culture growing on PDA, was inserted in a basal stem wound made with a sterile scalpel at 2 cm above ground level and sealed with Parafilm. Controls were inoculated with sterile PDA discs. There were 10 replicates for each isolate and host with an equal number of uninoculated plants. Seedlings were maintained in a greenhouse at 23 to 25°C. Within 10 to 15 days after inoculation, symptoms developed as water-soaked lesions followed by necrosis and finally wilting. The fungus was reisolated from the stems of all inoculated plants, completing Koch's postulates. To our knowledge, this is the first report of D. bryoniae in Tunisia. References: (1) A. P. Keinath et al. Phytopathology 85:364, 1995. (2) E. Punithalingam and P. Holliday. No. 332 in: Descriptions of Pathogenic Fungi and Bacteria. CMI, Kew, Surrey, UK, 1972.