Antifungal modes of action of metalaxyl, cyprofuram, benalaxyl and oxadixyl in phenylamide-sensitive and phenylamide-resistant strains of Phytophthora megasperma f. sp. medicaginis and Phytophthora infestans

Abstract

The activity of the phenylamide fungicides metalaxyl, cyprofuram, benalaxyl and oxadixyl against phenylamide-sensitive and phenylamide-resistant strains of Phytophthora megasperma f. sp. medicaginis and Phytophthora infestans has been compared in several tests. Metalaxyl showed the highest activity amongst the four fungicides against mycelial growth of sensitive strains on agar media. Benalaxyl and cyprofuram showed the highest activity against resistant strains. RNA synthesis of sensitive strains, measured as [ 3H]uridine incorporation, was inhibited by about 80% ( P. megasperma) and by about 40% ( P. infestans) by all phenylamides at concentrations of 1 μg/ml. At higher concentrations inhibition remained at this level, indicating that only part of the RNA synthesis was inhibited. RNA synthesis of resistant strains was completely insensitive to metalaxyl and oxadixyl at concentrations as high as 200 μg/ml. Cyprofuram, however, at concentrations above 1 μg/ml ( P. megasperma) and 10 μg/ml ( P. infestans) was inhibitory and at 200 μg/ml this compound almost completely inhibited the phenylamide-sensitive part of the RNA synthesis of both strains. In contrast to the other phenylamides, benalaxyl affected [ 3H]uridine uptake into mycelium at concentrations higher than 10 μg/ml, and at 100 μg/ml inhibition of uptake was almost total with both sensitive and resistant strains of both Phytophthora species. Inhibition of [ 3H]uridine uptake obviously is a feature of a second mechanism of action of benalaxyl. Endogenous RNA polymerase activity of isolated nuclei from sensitive strains of P. megasperma was inhibited at phenylamide concentrations as low as 0·1 μg/ml whereas that from resistant strains was not affected at concentrations as high as 100 μg/ml. Similarly, endogenous nuclear RNA polymerase activity of sensitive P. infestans strains appeared to be more sensitive to the phenylamides than that of resistant P. infestans strains. This suggests that resistance that developed in field strains of P. infestans has a basis similar to that of mutagen-induced resistance in laboratory strains of P. megasperma. Tests of the fungicidal activity of the phenylamides determined in a lucerne seedling assay ( P. megasperma) and a detached potato leaf assay ( P. infestans) clearly differentiated between sensitive and resistant strains. Resistance levels to cyprofuram and benalaxyl were as high as those to metalaxyl and oxadixyl, indicating that the additional effect of a second mechanism of action of benalaxyl and the property of cyprofuram to inhibit RNA synthesis of resistant strains at higher concentrations are of limited practical importance for counteracting phenylamide resistance.