title: Barley ROP binding kinase1 is involved in microtubule organization and in basal penetration resistance to the barley powdery mildew fungus.
abstract: Certain plant receptor-like cytoplasmic kinases were reported to interact with small monomeric G-proteins of the RHO of plant (ROP; also called RAC) family in planta and to be activated by this interaction in vitro. We identified a barley (Hordeum vulgare) partial cDNA of a ROP binding protein kinase (HvRBK1) in yeast (Saccharomyces cerevisiae) two-hybrid screenings with barley HvROP bait proteins. Protein interaction of the constitutively activated (CA) barley HvROPs CA HvRACB and CA HvRAC1 with full-length HvRBK1 was verified in yeast and in planta. Green fluorescent protein-tagged HvRBK1 appears in the cytoplasm and nucleoplasm, but CA HvRACB or CA HvRAC1 can recruit green fluorescent protein-HvRBK1 to the cell periphery. Barley HvRBK1 is an active kinase in vitro, and activity is enhanced by CA HvRACB or GTP-loaded HvRAC1. Hence, HvRBK1 might act downstream of active HvROPs. Transient-induced gene silencing of barley HvRBK1 supported penetration by the parasitic fungus Blumeria graminis f. sp. hordei, suggesting a function of the protein in basal disease resistance. Transient knockdown of HvRBK1 also influenced the stability of cortical microtubules in barley epidermal cells. Hence, HvRBK1 might function in basal resistance to powdery mildew by influencing microtubule organization.
authors.affiliation: Lehrstuhl für Phytopathologie, Technische Universität München, D-85350 Freising-Weihenstephan, Germany.
authors.forename: Christina
authors.initials: C
authors.lastname: Huesmann
authors.forename: Tina
authors.initials: T
authors.lastname: Reiner
authors.forename: Caroline
authors.initials: C
authors.lastname: Hoefle
authors.forename: Jutta
authors.initials: J
authors.lastname: Preuss
authors.forename: Manuela E
authors.initials: ME
authors.lastname: Jurca
authors.forename: Mónika
authors.initials: M
authors.lastname: Domoki
authors.forename: Attila
authors.initials: A
authors.lastname: Fehér
authors.forename: Ralph
authors.initials: R
authors.lastname: Hückelhoven
mesh_terms: D000595:Amino Acid Sequence; D001203:Ascomycota / Q000472:pathogenicity*; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D060467:Disease Resistance; D031541:Fluorescence Resonance Energy Transfer; D055785:Gene Knockdown Techniques; D020868:Gene Silencing; D017343:Genes, Plant; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D001467:Hordeum / Q000201:enzymology / Q000235:genetics / Q000382:microbiology*; D008870:Microtubules / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D010935:Plant Diseases / Q000276:immunology / Q000382:microbiology; D019441:Plant Epidermis / Q000235:genetics / Q000378:metabolism; D057865:Plant Immunity; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism*; D025941:Protein Interaction Mapping; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism
chemical_list: D018076:DNA, Complementary; D010940:Plant Proteins; D049452:Green Fluorescent Proteins; D017346:Protein Serine-Threonine Kinases