Plant pathogenic fungi secrete effector proteins and secondary metabolites to cause disease.Additionally, some produce small RNAs (sRNAs) that silence transcripts of host immunity genes through RNA interference. The fungus Sclerotinia sclerotiorum infects over 600 plant species, but little is known about its molecular interactions with its hosts. In particular, the role of sRNAs in S. sclerotiorum pathogenicity has not been determined. By sequencing sRNAs in vitro and during infection of two host species, we found that S. sclerotiorum produces at least 374 highly abundant sRNAs. These sRNAs mostly originated from polymorphic repeat-rich genomic regions. Predicted gene targets of these sRNAs, from 10 different host species, were enriched for immunity-related functional domains. Predicted A. thaliana gene targets of S. sclerotiorum sRNAs were significantly more down-regulated during infection than other genes. A. thaliana gene targets were also more likely to contain single nucleotide polymorphisms (SNPs) associated with quantitative disease resistance. In conclusion, sRNAs produced by S. sclerotiorum are likely capable of silencing immunity components in multiple hosts. Prediction of fungal sRNA targets in host plant genomes can be combined with other global approaches, such as genome wide association studies and transcriptomics, to assist identification of plant genes involved in disease resistance.

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