Effects of agricultural fungicides on microorganisms associated with floral nectar: susceptibility assays and field experiments

Pesticides have become an inseparable element of agricultural intensification. While the direct impact of pesticides on non-target organisms, such as pollinators, has recently received much attention, less consideration has been given to the microorganisms that are associated with them. Specialist yeasts and bacteria are known to commonly inhabit floral nectar and change its chemical characteristics in numerous ways, possibly influencing pollinator attraction. In this study, we investigated the in vitro susceptibility of nectar yeasts Metschnikowia gruessi, Metschnikowia reukaufii, and Candida bombi to six widely used agricultural fungicides (prothioconazole, tebuconazole, azoxystrobin, fenamidone, boscalid, and fluopyram). Next, a commercial antifungal mixture containing tebuconazole and trifloxystrobin was applied to natural populations of the plant Linaria vulgaris and the occurrence, abundance, and diversity of nectar-inhabiting yeasts and bacteria was compared between treated and untreated plants. The results showed that prothioconazole and tebuconazole were highly toxic to nectar yeasts, inhibiting their growth at concentrations varying between 0.06 and 0.5 mg/L. Azoxystrobin, fenamidone, boscalid, and fluopyram on the other hand exhibited considerably lower toxicity, inhibiting yeast growth at concentrations between 1 and 32 mg/L or in many cases not inhibiting microbial growth at all. The application of the antifungal mixture in natural plant populations resulted in a significant decrease in the occurrence and abundance of yeasts in individual flowers, but this did not translate into noticeable changes in bacterial incidence and abundance. Yeast and bacterial species richness and distribution did not also differ between treated and untreated plants. We conclude that the application of fungicides may have negative effects on the abundance of nectar yeasts in floral nectar. The consequences of these effects on plant pollination processes in agricultural systems warrant further investigation.