First Report of the Prevalence of Benzimidazole-Resistant Isolates in a Population of Cylindrocladium pauciramosum in Italy.

Abstract

Cylindrocladium pauciramosum C.L. Schoch & Crous (teleomorph Calonectria pauciramosa C.L. Schoch & Crous), described as a member of the Cylindrocladium candelabrum Viégas complex (4), was recently reported from Europe (3). In southern Italy, the fungus has caused extensive losses, and chemical control measures are necessary, especially in ornamental nurseries. Several researchers have found benzimidazole fungicides to be effective for control of different species of Cylindrocladium, however, in fungicide trials conducted on myrtle plants infected by C. pauciramosum, benomyl was ineffective (2). Another study showed that mycelial growth of six isolates was completely inhibited by carbendazim at a concentration of 1 μg a.i./ml whereas, concentrations of 10, 100, and 500 μg a.i./ml did not completely inhibit growth of four isolates (1). To examine benzimidazole resistance in C. pauciramosum, 200 single-conidia isolates were tested. These were collected during 1996 and 1997 from several symptomatic hosts in different nurseries located in Sicily and Calabria and identified through morphological characteristics as well as mating-type studies with tester strains. Sensitivity to benomyl was determined by plating mycelial plugs on potato dextrose agar (PDA) amended with benomyl at 1, 10, 100, and 500 μg a.i./ml. For 20 benomyl-resistant isolates, fungal growth was also determined at the same concentrations on carbendazim-amended PDA. Sensitivity was expressed as the minimum inhibitory concentration (MIC) (the lowest fungicide concentration that completely prevented fungal growth). Isolates that did not grow on benzimidazole-amended PDA were classed as sensitive. Isolates were considered resistant to benzimidazole if MIC values were greater than 1 μg a.i./ml. Of the 200 isolates tested, 58% were resistant to benomyl. The benomyl-resistant isolates tested for carbendazim sensitivity were cross-resistant to carbendazim. Most resistant isolates grew in the presence of benomyl at 500 μg a.i./ml. On agar culture, the isolates were either the fast-growing or slow-growing type. The slow-growing phenotype appears to be related to the higher level of resistance (500 μg a.i./ml). On the basis of these data, the use of benzimidazole for the control of this pathogen should be seriously questioned. To our knowledge, this is the first report of the prevalence of benzimidazole resistance in a population of C. pauciramosum.