Botrytis fragariae, a New Species Causing Gray Mold on Strawberries, Shows High Frequencies of Specific and Efflux-Based Fungicide Resistance

Botrytis cinerea causes pre- and postharvest decay of many fruit and vegetable crops. A survey in German strawberry fields revealed Botrytis strains that differed from B. cinerea in diagnostic PCR markers and growth appearance. Phylogenetic analyses showed these strains to belong to an undescribed species in Botrytis clade 2, named Botrytis fragariae sp. nov. Isolates of B. fragariae were detected in strawberry fields throughout Germany, sometimes at similar frequencies as B. cinerea , and in the Southeastern United States. B. fragariae was isolated from overwintering strawberry tissue, but not from freshly infected fruit. B. fragariae invaded strawberry tissues with similar or lower efficiency than B. cinerea but showed poor colonization of inoculated non-host plant tissues. These data and its exclusive occurrence on strawberry indicate that B. fragariae is host-specific and has a different tissue preference than B. cinerea . Various fungicide resistance patterns were observed in B. fragariae populations. Many B. fragariae strains showed resistance to one or several chemical classes of fungicides, and an efflux-based multidrug resistance (MDR1) phenotype previously described for B. cinerea . Resistance-related mutations in B. fragariae were identical or similar to those of B. cinerea for carbendazim (E198A mutation in tubA ), azoxystrobin (G143A in cytB ), iprodione (G367A+V368F in bos1 ) and MDR1 (gain-of-function mutations in the transcription factor mrr1 gene, and overexpression of the drug efflux transporter gene atrB ). The widespread occurrence of B. fragariae indicates that this species is adapted to fungicide-treated strawberry fields and may be of local importance as a gray mold pathogen alongside B. cinerea . Importance Gray mold is the most important fruit rot on strawberries worldwide, and requires fungicide treatments for control. For a long time it was believed to be caused only by Botrytis cinerea , a ubiquitous pathogen with broad host range which quickly develops fungicide resistance. We report the discovery and description of a new species, named Botrytis fragariae , which is widely distributed in commercial strawberry fields in Germany and the Southeastern United States. It was observed on overwintering tissue but not on freshly infected fruit and seems host-specific based on its occurrence and on artificial infection tests. B. fragariae has also developed resistance to several fungicides which is caused by similar mutations as those known for B. cinerea , including an efflux-based multidrug resistance. Our data indicate that B. fragariae could be of practical importance as a strawberry pathogen in some regions where it is similarly abundant as B. cinerea .