Assessment of Mycosphaerella graminicola resistance to azoxystrobin

Azoxystrobin resistance levels of twenty two strains sampled from ten French locations and two reference isolates (IPO323 and IPO94269) of the wheat pathogen Mycosphaerella graminicola were investigated in vitro. French strains assayed were selected from twenty two genetic groups determined from three hundred sixty three strains previously characterised using microsatellites, actine and beta-tubuline markers. For the first time, the evaluation was carried out using four distinct methods: spotting on PDA medium, spore germination on PDA medium and using microtitre plates with and without Alamar blue, a growth indicator. From dose-response curve, half maximal inhibitory concentration (IC50) was determined for each strain. The results obtained using microtitre plates with the addition of Alamar blue displayed high standard deviations from the growth averages observed. Therefore, we suggest that this method is inadequate to assess M. graminicolo resistance to fungicides. However, a good correlation was observed between the rankings of strains according to their IC50 values with the three other methods used. The two reference isolates, as expected, were inhibited by low azoxystrobin concentrations. On the other hand, the IC50 values obtained showed presence of a threshold between sensitive and resistant strains that corroborates the disruptive resistance of M. graminicola against strobilurin fungicides. In addition, the strains showing resistance were those sampled mainly from northern France, where a high frequency of strobilurin resistant isolates among M. graminicola populations was reported by several studies.