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Plant Molecular Biology

, Volume 65, Issue 1–2, pp 177–187 | Cite as

Elicitor induced activation of the methylerythritol phosphate pathway toward phytoalexins biosynthesis in rice

  • Atsushi Okada
  • Takafumi Shimizu
  • Kazunori OkadaEmail author
  • Tomohisa Kuzuyama
  • Jinichiro Koga
  • Naoto Shibuya
  • Hideaki Nojiri
  • Hisakazu Yamane
Article

Abstract

Diterpenoid phytoalexins such as momilactones and phytocassanes are produced via geranylgeranyl diphosphate in suspension-cultured rice cells after treatment with a chitin elicitor. We have previously shown that the production of diterpene hydrocarbons leading to phytoalexins and the expression of related biosynthetic genes are activated in suspension-cultured rice cells upon elicitor treatment. To better understand the elicitor-induced activation of phytoalexin biosynthesis, we conducted microarray analysis using suspension-cultured rice cells collected at various times after treatment with chitin elicitor. Hierarchical cluster analysis revealed two types of early-induced expression (EIE-1, EIE-2) nodes and a late-induced expression (LIE) node that includes genes involved in phytoalexins biosynthesis. The LIE node contains genes that may be responsible for the methylerythritol phosphate (MEP) pathway, a plastidic biosynthetic pathway for isopentenyl diphosphate, an early precursor of phytoalexins. The elicitor-induced expression of these putative MEP pathway genes was confirmed by quantitative reverse-transcription PCR. 1-Deoxy-d-xylulose 5-phosphate synthase (DXS), 1-deoxy-d-xylulose 5-phosphate reductoisomerase (DXR), and 4-(cytidine 5′-diphospho)-2-C-methyl-d-erythritol synthase (CMS), which catalyze the first three committed steps in the MEP pathway, were further shown to have enzymatic activities that complement the growth of E. coli mutants disrupted in the corresponding genes. Application of ketoclomazone and fosmidomycin, inhibitors of DXS and DXR, respectively, repressed the accumulation of diterpene-type phytoalexins in suspension cells treated with chitin elicitor. These results suggest that activation of the MEP pathway is required to supply sufficient terpenoid precursors for the production of phytoalexins in infected rice plants.

Keywords

Disease resistance Diterpenoid phytoalexins Elicitor MEP pathway Oryza sativa Rice 

Notes

Acknowledgements

We thank Dr Yoshiaki Nagamura and Ms Ritsuko Motoyama of the Rice Genome Resource Center for technical support with the microarray analysis, and also for providing the rice full-length cDNA clone that was developed in the Rice Genome Project of the National Institute of Agrobiological Sciences, Japan, and Prof Tadao Asami in The University of Tokyo for distribution of 5-ketoclomazone. This work was supported by the Program for Promotion of Basic Research Activities for Innovative Biosciences (PROBRAIN).

Supplementary material

11103_2007_9207_MOESM1_ESM.xls (195 kb)
List of genes categorized in the EIE-1 node (XLS 195 KB)
11103_2007_9207_MOESM2_ESM.xls (102 kb)
List of genes categorized in the EIE-2 node (XLS 103 KB)
11103_2007_9207_MOESM3_ESM.xls (80 kb)
List of genes categorized in the LIE node (XLS 80 KB)
11103_2007_9207_MOESM4_ESM.doc (26 kb)
List of microarray data of the MVA pathway gene homologs in rice (DOC 27 KB)
11103_2007_9207_MOESM5_ESM.doc (24 kb)
List of microarray data of the DXS homologs in rice (DOC 25 KB)

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Copyright information

© Springer Science+Business Media B.V. 2007

Authors and Affiliations

  • Atsushi Okada
    • 1
  • Takafumi Shimizu
    • 1
  • Kazunori Okada
    • 1
    Email author
  • Tomohisa Kuzuyama
    • 1
  • Jinichiro Koga
    • 2
  • Naoto Shibuya
    • 3
  • Hideaki Nojiri
    • 1
  • Hisakazu Yamane
    • 1
  1. 1.Biotechnology Research CenterThe University of TokyoTokyoJapan
  2. 2.Food and Health R&D LaboratoriesMeiji Seika Kaisha, Ltd.Sakado-shiJapan
  3. 3.Department of Life Sciences, Faculty of AgricultureMeiji UniversityKawasakiJapan

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