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Development and Significance of Dicarboximide Resistance in Sclerotinia minor Isolates from Commercial Lettuce Fields in California

Three growth stages of each of 20 Sclerotinia minor isolates were tested for resistance to iprodione. Sclerotia and both vegetative and mature mycelium of each isolate were plated on potato-dextrose agar (PDA) amended with 0, 1, 5, 10, 25, and 100 µg of the fungicide per ml, and radial growth was measured. All wild-type field isolates were initially sensitive and did not grow in the presence of iprodione. However, fungicide resistance arose readily in vitro. All 20 isolates produced measurable growth (≥2 mm) on iprodione at 5 µg/ml after 2 weeks when started from mature mycelium, and 18 of 20 isolates produced measurable growth after 4 weeks when started from vegetative mycelium. In general, fungicide-resistant growth arose more frequently and mean colony diameters were significantly greater (P ≤ 0.05) with mature mycelium than with vegetative mycelium at all times and concentrations. In sclerotial germination tests, at least 1% of sclerotia germinated in 18 of 20 isolates after 2 weeks on iprodione at 5 µg/ml, and in 19 of 20 isolates after 5 weeks on 100 µg/ml. Of growth produced on 79 plates containing iprodione, 73 remained viable on PDA after 5 months, and 71 retained resistance to 5 µg/ml. Seventy of these also exhibited cross-resistance to vinclozolin, another dicarboximide fungicide. Pathogenicity of five fungicide-resistant strains was tested in greenhouse, microplot, and field experiments, with and without iprodione. Two months after in vitro production, one of the five resistant strains was avirulent, but disease incidence for the other four ranged from 40 to 75%, compared with 40% for the wild-type isolates. However, the virulence of the fungicide-resistant strains declined over time. Ten months after their production, two of the isolates were avirulent and disease incidence for the others ranged from 3 to 17%, compared with 40 to 90% for the wild-type isolates. The strains that remained virulent 5 and 7 months after in vitro production were not significantly controlled by iprodione sprayed according to labeled rates, although disease was significantly controlled by the fungicide in treatments inoculated with wild-type field isolates (P > 0.05). In experiments in commercial fields to determine the efficacy of fungicide sprays on the wild-type S. minor population, there was no evidence that fungicide resistance was the cause of lack of lettuce drop control observed in many coastal California fields. Application of fungicides at a less than optimal time may account for some incidents of control failure.