Role of jasmonate in the rice (Oryza sativa L.) self-defense mechanism using proteome analysis

Electrophoresis. 2000 Jul;21(12):2492-500. doi: 10.1002/1522-2683(20000701)21:12<2492::AID-ELPS2492>3.0.CO;2-2.

Abstract

Exogenously applied jasmonic acid (JA) was used to study changes in protein patterns in rice (Oryza sativa L.) seedling tissues, to classify these changes, and to assign a role for these changes, in order to define the role of JA in the rice self-defense mechanism. High resolution two-dimensional polyacrylamide gel electrophoretic analysis revealed induction of new proteins in both leaf and stem tissues after JA treatment, with the major protein spots further analyzed through N-terminal and internal amino acid sequencing, purification, antibody production, and immunoblot analysis. JA treatment results in necrosis in these tissues, which is accompanied by drastic reductions in ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) subunits, and was confirmed using immunoblotting. Induction of novel proteins was found particularly in the stem tissues, including a new basic 28 kDa Bowman-Birk proteinase inhibitor protein (BBPIN; jasmonate-induced stem protein, JISP 6), and acidic 17 kDa pathogenesis-related class 1 protein (PR-1, JISP 9). This induction of proteins was blocked by a protein synthesis inhibitor cycloheximide (CHX) indicating de novo protein synthesis. Kinetin (KIN), a cytokinin and free radical scavenger reversed RuBisCO decreases, but not induction of proteins. Immunoblot analysis using antibodies generated against these purified proteins revealed a tissue-specific expression pattern and time-dependent induction after JA treatment. Our results indicate that jasmonate affects defense-related gene expression in rice seedlings, as evidenced by de novo synthesis of novel proteins with potential roles in plant defense.

MeSH terms

  • Amino Acids / analysis
  • Cyclopentanes / metabolism*
  • Histone-Lysine N-Methyltransferase / biosynthesis
  • Immunoblotting / methods
  • Oryza / metabolism*
  • Oxylipins
  • Plant Growth Regulators / metabolism*
  • Plant Leaves / chemistry
  • Plant Proteins / biosynthesis
  • Plant Proteins / metabolism*
  • Plant Stems / chemistry
  • Proteome / metabolism*
  • Seeds / chemistry
  • Sequence Analysis, Protein
  • Trypsin Inhibitor, Bowman-Birk Soybean / biosynthesis
  • Trypsin Inhibitors / biosynthesis

Substances

  • Amino Acids
  • Cyclopentanes
  • Oxylipins
  • Plant Growth Regulators
  • Plant Proteins
  • Proteome
  • Trypsin Inhibitor, Bowman-Birk Soybean
  • Trypsin Inhibitors
  • pathogenesis-related proteins, plant
  • jasmonic acid
  • ribulose-1,5-bisphosphate carboxylase-oxygenase large subunit epsilonN-methyltransferase
  • Histone-Lysine N-Methyltransferase