Sensitivity and cross-resistance patterns to demethylation-inhibiting fungicides in California populations of Alternaria alternata pathogenic on pistachio

Alternaria late blight (ALB) caused by Alternaria spp. is an annual production concern for pistachio growers in California. In this study, isolates of Alternaria alternata collected from California pistachio orchards between 1998 and 2003 (population A, n = 46) and in 2010 (population B, n = 38) prior to the registration and use of sterol demethylation-inhibiting (DMI) fungicides for ALB control, were tested for their sensitivity to difenoconazole, propiconazole, and tebuconazole using an in vitro mycelial growth assay. The 50% effective dose (EC₅₀) values for isolates in population A to difenoconazole, propiconazole, and tebuconazole, respectively, ranged from 0.02 to 0.82 μg/mL (mean = 0.17), 0.25–5.73 μg/mL (mean = 1.29), and 0.02–2.76 μg/mL (mean = 0.96), and from 0.02 to 0.93 μg/mL (mean = 0.21), 0.13–5.36 μg/mL (mean = 1.25), and 0.17–5.47 μg/mL (mean = 1.03) for those in population B. Among 65 isolates collected in 2012 from orchards with very limited exposure to difenoconazole, propiconazole or metconazole (population C, n = 65), the EC₅₀ values for tebuconazole ranged from 0.02 to 2.99 μg/mL (mean = 0.51), while EC₅₀ values for difenoconazole of 55 of these isolates varied from 0.01 to 0.44 μg/mL (mean = 0.06). The DMI-exposed population C was found to be on average two to four times less sensitive compared with the populations A and B. Pearson correlation analyses of EC₅₀ values for the three fungicides showed significant positive correlations between the sensitivities of tebuconazole and propiconazole (P < 0.01, r = 0.32), tebuconazole and difenoconazole (P < 0.01, r = 0.26), and difenoconazole and propiconazole (P < 0.01, r = 0.40). Results indicate a clear difference in fungicide intrinsic activity, with difenoconazole being more active than the two other DMIs. Although wide variations in sensitivities for tebuconazole and propiconazole were observed in the non- and DMI-exposed Alternaria populations, efficacy of DMI products has been excellent against ALB. The data collected here will be crucial for the rational use of DMIs in fungicide spray programs for ALB management and serve as a reference to detect any shifts in A. alternata sensitivity to these fungicides in subsequent years as they become more frequently used in California pistachios.