Differential Expression of Defense/Stress-Related Marker Proteins in Leaves of a Unique Rice Blast Lesion Mimic Mutant (blm)

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• 作者：Young-Ho Jung ; Randeep Rakwal ; Ganesh Kumar Agrawal ; Junko Shibato ; Jung-A Kim ; Mi Ok Lee ; Pil-Kyu Choi ; Seung-Hee Jung ; So Hee Kim ; Hee-Jong Koh ; Masami Yonekura ; Hitoshi Iwahashi ; Nam-Soo Jwa
• 刊名：Journal of Proteome Research
• 出版年：2006
• 出版时间：October 2006
• 年：2006
• 卷：5
• 期：10
• 页码：2586 - 2598
• 全文大小：817K
• 年卷期：v.5,no.10(October 2006)
• ISSN：1535-3907

文摘

We analyzed a unique rice (Oryza sativa L.) blast lesion mimic (blm) mutant for differentially expressedproteins 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 andwild-type, WT) before (stage 1, S1) and after (stage 2, S2) lesion formation. 1-DGE immunoblottingrevealed potent increase in the expression of a key pathogenesis-related (PR) marker biosyntheticenzyme, naringenin 7-O-methyltransferase, involved in rice phytoalexin sakuranetin biosynthesis, andthree oxidative-stress-related marker proteins, catalase, ascorbate peroxidase (APX), and superoxidedismutase (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 detected50 Coomassie brilliant blue-stained protein spots differentially expressed in blm. A total of 23 and 44protein spots was excised for analysis by N-terminal amino acid sequencing and nano-electrosprayionization liquid chromatography mass spectrometry, respectively; 26 nonredundant proteins wereidentified. The pathogenesis-related class 5 and 10 proteins, including a new OsPR10d protein, weresignificantly induced in blm. The OsPR5 protein spot was stained with Pro-Q Diamond phosphoproteingel stain suggesting OsPR5 to be a putative phosphoprotein. Surprisingly, protein spot 20, a leafOsPR10b, showed identity to a rice root-specific PR-10 (RSOsPR10). To resolve this discrepancy, wechecked its expression in leaves of blm and WT (S1 and S2), respectively, using gene-specific primersand reverse transcriptase-polymerase chain reaction; RSOsPR10 mRNA was found to express in theleaves.