First Report of Thiabendazole-Resistant Isolates of Fusarium sambucinum Infecting Stored Potatoes in Nova Scotia, Canada.

Abstract

Fusarium dry rot is a significant postharvest disease of potato (Solanum tuberosum L.) and is often controlled by applying thiabendazole to tubers prior to storage. However, thiabendazole-resistant isolates of Fusarium spp. have been reported from Europe (2), the United States (1), and Canada (1,4). To address concerns, samples of potato tubers showing symptoms of dry rot caused by Fusariumspp. were collected from three storage bays in a commercial storage facility in Nova Scotia, Canada, in February 2001. All tubers had been treated with thiabendazole after harvest and prior to storage. Tubers were cut longitudinally, and small tissue samples (10 × 5 × 3 mm) were taken from the margins of internal necrotic regions with a sterile scalpel, surface-sterilized in 0.6% sodium hypochlorite for 15 s, rinsed twice in sterile distilled water (SDW), and blotted dry on sterile filter paper. Tissue pieces were plated on 0.5-strength potato dextrose agar (PDA) amended with tetracycline (0.05 g/liter) and streptomycin sulfate (0.1 g/liter). Petri dishes were incubated in the dark at 22°C for 4 to 7 days. After incubation, hyphal tips from the margins of actively growing isolates were removed with a sterile probe and plated on 0.5-strength PDA to generate pure cultures. Of 35 potato tubers examined, 10 (29%) yielded Fusarium isolates for further study. All 10 isolates were identified as F. sambucinum Fuckel according to Nelson et al. (3). Agar plugs (5 mm diameter) taken from the margins of 7- to 10-day-old cultures of F. sambucinum isolates were transferred to petri dishes containing 0.5-strength PDA amended with thiabendazole at 0, 1, 5, 10, 20, 50, or 100 mg/liter. Thiabendazole was prepared as a stock solution in SDW and added to molten agar after autoclaving. Cultures were grown in the dark for 7 days at 22°C, after which mycelial growth diameter was measured using digital calipers. Two measurements, along orthogonal diameters, were taken from each of three replicate plates for a total of six measurements per thiabendazole concentration. Means were calculated, and the diameter of the inoculation plug was subtracted from each mean. Calculated EC50 values (thiabendazole concentration inhibiting pathogen growth by 50%) were obtained by regression of the log of the chemical concentration against the corresponding probit of percent fungal inhibition. All isolates of F. sambucinum were resistant to thiabendazole, with EC50 values ranging from 7 to 82 mg/liter. Six isolates had EC50 values between 40 and 82 mg/liter. Control isolates of F. sambucinum, F. avenaceum, F. solani, and F. oxysporum were sensitive to thiabendazole, with EC50 values of <1 mg/liter. Although isolates of F. sambucinum resistant to thiabendazole have been recovered from eastern Canada (1,4), this is the first report of thiabendazole resistance in F. sambucinum isolates from tubers in commercial storage in the Annapolis Valley of Nova Scotia, Canada, a production region that concentrates on growing processing potatoes for the potato chip industry and is several hundred kilometers from other potato-growing regions of Prince Edward Island and New Brunswick.