title: Increased resistance to crown rust disease in transgenic Italian ryegrass (Lolium multiflorum Lam.) expressing the rice chitinase gene.
abstract: We introduced the rice chitinase (Cht-2; RCC2) gene into calli of Italian ryegrass (Lolium multiflorum Lam.), with a hygromycin phosphotransferase (HPT) gene as a selectable marker, by particle bombardment. Hygromycin-resistant calli were selected and transferred to regeneration medium for shoot formation. Polymerase chain reaction (PCR) analysis revealed regenerants containing the HPT gene. The RCC2 gene was detected in 65.5% of those regenerants. Southern hybridization detected both HPT and RCC2 genes and indicated that the transgenic plants were independently transformed. Expression of the RCC2 gene in the transgenic plants was confirmed by Northern hybridization, reverse transcription-PCR and Western blotting. Bioassay of detached leaves indicated increased resistance to crown rust (Puccinia coronata) in transgenic plants, which exhibited higher chitinase activity than a nontransgenic plant.
authors.affiliation: Forage Crop Research Institute, Japan Grassland Farming and Forage Seed Association, 388-5 Higashiakada, Nishinasuno, Tochigi, 329-2742, Japan. takahashi@jfsass.or.jp
authors.forename: Wataru
authors.initials: W
authors.lastname: Takahashi
authors.forename: Masahiro
authors.initials: M
authors.lastname: Fujimori
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Miura
authors.forename: Toshinori
authors.initials: T
authors.lastname: Komatsu
authors.forename: Yoko
authors.initials: Y
authors.lastname: Nishizawa
authors.forename: Tadaaki
authors.initials: T
authors.lastname: Hibi
authors.forename: Tadashi
authors.initials: T
authors.lastname: Takamizo
mesh_terms: D000383:Agriculture / Q000379:methods; D019470:Biolistics; D002688:Chitinases / Q000235:genetics*; D018506:Gene Expression Regulation, Plant / Q000235:genetics; D018014:Gene Transfer Techniques; D005819:Genetic Markers / Q000235:genetics; D007113:Immunity, Innate / Q000235:genetics*; D008129:Lolium / Q000201:enzymology / Q000235:genetics* / Q000382:microbiology; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics; D010935:Plant Diseases / Q000235:genetics*; D010940:Plant Proteins / Q000235:genetics*; D030821:Plants, Genetically Modified / Q000201:enzymology / Q000235:genetics* / Q000382:microbiology; D012038:Regeneration / Q000235:genetics; D014170:Transformation, Genetic / Q000235:genetics; D019076:Transgenes / Q000235:genetics
chemical_list: D005819:Genetic Markers; D010940:Plant Proteins; C092515:RCg2 protein, Oryza sativa; D017853:Phosphotransferases (Alcohol Group Acceptor); C063242:hygromycin-B kinase; D002688:Chitinases