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Although barley (<jats:italic>Hordeum vulgare<\/jats:italic>) <jats:italic>mlo<\/jats:italic> mutants have been used in agriculture for more than 40\u2009years, understanding of the molecular principles underlying this type of disease resistance remains fragmentary. Forward genetic screens in barley have revealed mutations in two <jats:italic>Required for mlo resistance<\/jats:italic> (<jats:italic>Ror<\/jats:italic>) genes that partially impair immunity conferred by <jats:italic>mlo<\/jats:italic> mutants. While <jats:italic>Ror2<\/jats:italic> encodes a soluble <jats:italic>N<\/jats:italic>\u2010ethylmaleimide\u2010sensitive factor\u2010attached protein receptor (SNARE), the identity of <jats:italic>Ror1<\/jats:italic>, located at the pericentromeric region of barley chromosome 1H, remained elusive. We report the identification of <jats:italic>Ror1<\/jats:italic> based on combined barley genomic sequence information and transcriptomic data from <jats:italic>ror1<\/jats:italic> mutant plants. <jats:italic>Ror1<\/jats:italic> encodes the barley class XI myosin Myo11A (HORVU.MOREX.r3.1HG0046420). Single amino acid substitutions of this myosin, deduced from non\u2010functional <jats:italic>ror1<\/jats:italic> mutant alleles, map to the nucleotide\u2010binding region and the interface between the relay\u2010helix and the converter domain of the motor protein. Ror1 myosin accumulates transiently in the course of powdery mildew infection. Functional fluorophore\u2010labeled Ror1 variants associate with mobile intracellular compartments that partially colocalize with peroxisomes. Single\u2010cell expression of the Ror1 tail region causes a dominant\u2010negative effect that phenocopies <jats:italic>ror1<\/jats:italic> loss\u2010of\u2010function mutants. We define a myosin motor for the establishment of <jats:italic>mlo<\/jats:italic>\u2010mediated resistance, suggesting that motor protein\u2010driven intracellular transport processes are critical for extracellular immunity, possibly through the targeted transfer of antifungal and\/or cell wall cargoes to pathogen contact sites.<\/jats:p>","DOI":"10.1111\/tpj.15930","type":"journal-article","created":{"date-parts":[[2022,8,2]],"date-time":"2022-08-02T19:34:53Z","timestamp":1659468893000},"page":"84-103","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":12,"title":["Barley <i>Ror1<\/i> encodes a class <scp>XI<\/scp> myosin required for <i>mlo<\/i>\u2010based broad\u2010spectrum resistance to the fungal powdery mildew 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