Differential expression of defense/stress-related marker proteins in leaves of a unique rice blast lesion mimic mutant (blm).
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.
Jung
Young-Ho
YH
Department of Molecular Biology, College of Natural Science, Sejong University, Seoul 143-747, Korea.
Rakwal
Randeep
R
Agrawal
Ganesh Kumar
GK
Shibato
Junko
J
Kim
Jung-A
JA
Lee
Mi Ok
MO
Choi
Pil-Kyu
PK
Jung
Seung-Hee
SH
Kim
So Hee
SH
Koh
Hee-Jong
HJ
Yonekura
Masami
M
Iwahashi
Hitoshi
H
Jwa
Nam-Soo
NS
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
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