title: Differential expression of defense/stress-related marker proteins in leaves of a unique rice blast lesion mimic mutant (blm).
abstract: We analyzed a unique rice (Oryza sativa L.) blast lesion mimic (blm) mutant for differentially expressed proteins in leaves of one- and two-week-old seedlings manifesting the lesion mimic phenotype. Gel-based one- and two-dimensional electrophoresis (1- and 2-DGE) was performed using leaves (blm and wild-type, WT) before (stage 1, S1) and after (stage 2, S2) lesion formation. 1-DGE immunoblotting revealed potent increase in the expression of a key pathogenesis-related (PR) marker biosynthetic enzyme, naringenin 7-O-methyltransferase, involved in rice phytoalexin sakuranetin biosynthesis, and three oxidative-stress-related marker proteins, catalase, ascorbate peroxidase (APX), and superoxide dismutase (SOD) in leaves of the blm mutant. 2-D gel immunoblotting analysis with anti-APX and anti-SOD antibodies revealed newly appearing cross-reacting protein spots in blm. 2-DGE analysis detected 50 Coomassie brilliant blue-stained protein spots differentially expressed in blm. A total of 23 and 44 protein spots was excised for analysis by N-terminal amino acid sequencing and nano-electrospray ionization liquid chromatography mass spectrometry, respectively; 26 nonredundant proteins were identified. The pathogenesis-related class 5 and 10 proteins, including a new OsPR10d protein, were significantly induced in blm. The OsPR5 protein spot was stained with Pro-Q Diamond phosphoprotein gel stain suggesting OsPR5 to be a putative phosphoprotein. Surprisingly, protein spot 20, a leaf OsPR10b, showed identity to a rice root-specific PR-10 (RSOsPR10). To resolve this discrepancy, we checked its expression in leaves of blm and WT (S1 and S2), respectively, using gene-specific primers and reverse transcriptase-polymerase chain reaction; RSOsPR10 mRNA was found to express in the leaves.
authors.affiliation: Department of Molecular Biology, College of Natural Science, Sejong University, Seoul 143-747, Korea.
authors.forename: Young-Ho
authors.initials: YH
authors.lastname: Jung
authors.forename: Randeep
authors.initials: R
authors.lastname: Rakwal
authors.forename: Ganesh Kumar
authors.initials: GK
authors.lastname: Agrawal
authors.forename: Junko
authors.initials: J
authors.lastname: Shibato
authors.forename: Jung-A
authors.initials: JA
authors.lastname: Kim
authors.forename: Mi Ok
authors.initials: MO
authors.lastname: Lee
authors.forename: Pil-Kyu
authors.initials: PK
authors.lastname: Choi
authors.forename: Seung-Hee
authors.initials: SH
authors.lastname: Jung
authors.forename: So Hee
authors.initials: SH
authors.lastname: Kim
authors.forename: Hee-Jong
authors.initials: HJ
authors.lastname: Koh
authors.forename: Masami
authors.initials: M
authors.lastname: Yonekura
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Iwahashi
authors.forename: Nam-Soo
authors.initials: NS
authors.lastname: Jwa
mesh_terms: D000595:Amino Acid Sequence; D060387:Ascorbate Peroxidases; D015415:Biomarkers / Q000032:analysis; D002374:Catalase / Q000032:analysis / Q000235:genetics; D015180:Electrophoresis, Gel, Two-Dimensional; D004734:Energy Metabolism; D005419:Flavonoids / Q000378:metabolism; D006360:Heat-Shock Proteins / Q000032:analysis* / Q000235:genetics; D008780:Methyltransferases / Q000032:analysis; D008969:Molecular Sequence Data; D009154:Mutation; D012275:Oryza / Q000235:genetics / Q000378:metabolism*; D018384:Oxidative Stress / Q000235:genetics; D010544:Peroxidases / Q000032:analysis / Q000235:genetics; D010750:Phosphoproteins / Q000032:analysis / Q000235:genetics; D010788:Photosynthesis; D010935:Plant Diseases / Q000235:genetics; D018515:Plant Leaves / Q000235:genetics / Q000378:metabolism*; D010940:Plant Proteins / Q000032:analysis* / Q000235:genetics; D020543:Proteome / Q000032:analysis* / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D012717:Sesquiterpenes; D013482:Superoxide Dismutase / Q000032:analysis / Q000235:genetics; D013729:Terpenes / Q000378:metabolism; D000094162:Phytoalexins
chemical_list: D015415:Biomarkers; D005419:Flavonoids; D006360:Heat-Shock Proteins; D010750:Phosphoproteins; D010940:Plant Proteins; D020543:Proteome; D012333:RNA, Messenger; D012717:Sesquiterpenes; D013729:Terpenes; C099724:sakuranetin; D010544:Peroxidases; D060387:Ascorbate Peroxidases; D002374:Catalase; D013482:Superoxide Dismutase; D008780:Methyltransferases; C099725:naringenin 7-O-methyltransferase; D000094162:Phytoalexins