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

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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.

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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).

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Authors and Affiliations

  1. Biotechnology Research Center, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo, 113-8657, Japan

    Atsushi Okada, Takafumi Shimizu, Kazunori Okada, Tomohisa Kuzuyama, Hideaki Nojiri & Hisakazu Yamane

  2. Food and Health R&D Laboratories, Meiji Seika Kaisha, Ltd., 5-3-1, Chiyoda, Sakado-shi, Saitama, 350-0289, Japan

    Jinichiro Koga

  3. Department of Life Sciences, Faculty of Agriculture, Meiji University, 1-1-1 Higashi-mita, Tama-ku, Kawasaki, Kanagawa, 214-8571, Japan

    Naoto Shibuya

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  1. Atsushi Okada
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  2. Takafumi Shimizu
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  3. Kazunori Okada
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  4. Tomohisa Kuzuyama
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  5. Jinichiro Koga
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Correspondence to Kazunori Okada.

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Okada, A., Shimizu, T., Okada, K. et al. Elicitor induced activation of the methylerythritol phosphate pathway toward phytoalexins biosynthesis in rice. Plant Mol Biol 65, 177–187 (2007). https://doi.org/10.1007/s11103-007-9207-2

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