show Abstracthide AbstractWe studied five isolates putatively from a clonal lineage of Puccinia hordei, the pathogen causing barley leaf rust. We de novo assembled a high quality genome for this pathogen using PacBio SMRT sequencing. A RNA-Seq of the pathogen during infection stages was performed and was used to guide gene annotation. Then, Illumina re-sequencing was performed for all isolates in order to call genotypes in relation to their phenotypes of virulence.