Resistance to pyraclostrobin, boscalid and multiple resistance to Pristine® (pyraclostrobin plus boscalid) fungicide in Alternaria alternata causing alternaria late blight of pistachios in California

Pristine (R) (pyraclostrobin + boscalid) is a fungicide registered for the control of alternaria late blight in pistachio. A total of 95 isolates of Alternaria alternata collected from orchards with and without a prior history of Pristine (R) sprays were tested for their sensitivity towards pyraclostrobin, boscalid and Pristine (R) in conidial germination assays. The EC50 values for 35 isolates from orchards without Pristine (R) sprays ranged from 0.09 to 3.14 mu g mL(-1) and < 0.01 to 2.04 mu g mL(-1) for boscalid and Pristine (R), respectively. For pyraclostrobin, 27 isolates had EC50 < 0.01 mu g mL(-1) and six had low resistance (mean EC50 value = 4.71 mu g mL(-1)). Only one isolate was resistant to all three fungicides tested, with EC50 > 100 mu g mL(-1). Among 59 isolates from the orchard with a history of Pristine (R) sprays, 56 were resistant to pyraclostrobin; only two were sensitive (EC50 < 0.01 mu g mL(-1)) and one was weakly resistant (EC50 = 10 mu g mL(-1)). For the majority of these isolates EC50 values ranged from 0.06 to 4.22 mu g mL(-1) for boscalid and from 0.22 to 7.74 mu g mL(-1) for Pristine (R). However, seven isolates resistant to pyraclostrobin were also highly resistant to boscalid and Pristine (R) and remained pathogenic on pistachio treated with Pristine (R). Whereas strobilurin resistance is a common occurrence in Alternaria of pistachio, this is the first report of resistance to boscalid in field isolates of phytopathogenic fungi. No cross resistance between pyraclostrobin and boscalid was detected, suggesting that Pristine (R) resistance appears as a case of multiple resistance.