Proteomic analysis of differentially expressed proteins induced by rice blast fungus and elicitor in suspension‐cultured rice cells

Abstract

We used two-dimensional electrophoresis (2-DE) and other proteomic approaches to identify proteins expressed in suspension-cultured rice cells in response to the rice blast fungus, Magnaporthe grisea. Proteins were extracted from suspension-cultured cells at 24 and 48 h after rice blast fungus inoculation or treatment with elicitor or other signal molecules such as jasmonic acid (JA), salicylic acid, and H(2)O(2). The proteins were then polyethylene glycol fractionated before separation by 2-DE. Fourteen protein spots were induced or increased by the treatments, which we analyzed by N-terminal or internal amino acid sequencing. Twelve proteins from six different genes were identified. Rice pathogen-related protein class 10 (OsPR-10), isoflavone reductase like protein, beta-glucosidase, and putative receptor-like protein kinase were among those induced by rice blast fungus; these have not previously been reported in suspension-cultured rice cells. Six isoforms of probenazole-inducible protein (PBZ1) and two isoforms of salt-induced protein (SalT) that responded to blast fungus, elicitor, and JA were also resolved on a 2-DE gel and identified by proteome analysis. The expression level of these induced proteins both in suspension-cultured cells and in leaves of whole plants was analyzed by Western blot. PBZ1, OsPR-10, and SalT proteins from incompatible reactions were induced earlier and to a greater extent than those in compatible reactions. Proteome analysis can thus distinguish differences in the timing and amount of protein expression induced by pathogens and other signal molecules in incompatible and compatible interactions.

Exogenous Application of Jasmonic Acid Induces Volatile Emissions in Rice and Enhances Parasitism of Nilaparvata lugens Eggs by theParasitoid Anagrus nilaparvatae

“…The fact that this difference was only apparent 1 hr after release suggests that JA-induced rice volatiles were slightly attractive to female N. lugens, whereas JA-induced nonvolatile chemicals had no particular effect on host preference. The latter is somewhat surprising as JA elicitation increases the production of proteinase inhibitors, phytoalexins, and PRs in rice plants (Rakwal and Komatsu, 2000;Rakwal et al, 2001;Kim et al, 2003). N. lugens female adults are attracted to limonene (Lou et al, unpublished data).…”

“…In rice, the world's most important food crop, the role of JA signaling has been mainly studied for direct defenses. Exogenous application of JA on rice plants elicits the productions of proteinase inhibitors, phytoalexins, PRs, and salt-induced proteins (Tamogamia et al, 1997;Rakwal and Komatsu, 2000;Rakwal et al, 2001;Kim et al, 2003), and it may increase the emission of volatiles (Obara et al, 2002).…”

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“…Additionally, five proteins (D1, D6-D8 and D10) appeared only on the control gel. In a previous report, proteomic analysis of rice suspension culture cells treated with rice blast fungus 25 also detected a 31.1 kDa (pI 4.98) protein, which was identified as putative receptor-like kinase. The same author also found an increase in the amount of 33.8 kDa proteins with pI of 4.73 and 4.79, which was identified as salt induced protein, SalT.…”

Gene Expression Profiling in Rice Infected with Rice Blast Fungus using SAGE

“…In general, both beta-glucanases and peroxidases have been known as enzymes that mediate the defense system against abiotic stresses such as ozone, H 2 O 2 and ultraviolet radiation (Cheong et al, 2000;Dabir et al, 2007;Kim et al, 2008b;Thalmair et al, 1996). The tags for PR-10 ( Kim et al, 2003;McGee et al, 2001;Tanaka et al, 2003) were detected a total of three times, suggesting that the protein seems to make a significant contribution as a biotic stress defense system. Recently, it was reported that the JIOsPR10 protein which is one of the PR-10 family possesses RNase activity that is dependent on sensitive to DTT, suggesting the importance of the disulfide bonding between cysteine residues (Kim et al, 2008a).…”