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Evidence for Methylerythritol Pathway (MEP) Contributions to Zerumbone Biosynthesis as Revealed by Expression Analysis of Regulatory Genes and Metabolic Inhibitors Studies

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Abstract

Sesquiterpenoid Zerumbone, the principal secondary metabolite in Zingiber zerumbet Smith, has been identified as the putative molecule conferring resistance against soft-rot causative, Pythium myriotylum. Metabolic precursors for sesquiterpenoid biosynthesis namely, isopentenyl diphosphate (IPP) and DMAPP, are generated either from cytosolic mevalonic acid (MVA) and/or the plastidal methylerythritol phosphate (MEP) pathway. Evaluation of expression pattern of regulatory genes of MEP and MVA pathway following P. myriotylum infection revealed that while transcripts of MVA regulatory gene, 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGR), did not show significant changes, biphasic expression pattern was observed for MEP pathway regulatory genes, viz., 1-deoxy-D-xylulose-5-phosphate reductase (DXR) and 1-deoxy-D-xylulose-5-phosphate synthase (DXS) as well for ZzTPS (terpene synthase) and Farnesyl phosphate synthase (FPS) compared with uninfected control. Contribution of the two pathways in zerumbone biosynthesis was evaluated in axenic tissue culture raised Z. zerumbet plantlets using MVA and MEP pathway specific inhibitors, mevinolin (MEV) and fosmidomycin (FOS) respectively. Results generated by regulatory gene and metabolite analysis are informative with respect to the role of plastidial IPP pool generated via MEP pathway in zerumbone biosynthesis and the possible redirection of flux in the event of metabolic perturbations mediated by chemical inhibitors.

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Acknowledgments

KD acknowledges the research fellowship received from Ministry of Human Resource and Development (MHRD), Government of India. Authors are grateful to NITC CUK for the research facilities extended.

Funding

The study was financially supported by the Department of Science and Technology-Science and Engineering Research Board (DST-SERB) for research grant (no. EMR/2016/002229).

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Central University of Kerala (CUK), Kasaragod, 671 320, India

    Aswati Ravindranathan Nair

  2. School of Biotechnology, National Institute of Technology Calicut, Calicut, 673 601, India

    Keerthi Divakaran

  3. Department of Genomic Science, CUK, Kasaragod, India

    Padmesh Pillai

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  1. Aswati Ravindranathan Nair
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  2. Keerthi Divakaran
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  3. Padmesh Pillai
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Contributions

RAN conceived and designed the experiments. RAN and KD performed the experiment. RAN, KD, and PP analyzed the results and wrote the manuscript. All authors have read and approved the final manuscript.

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Correspondence to Aswati Ravindranathan Nair.

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Key Message

• Biphasic expression observed for methylerythritol (MEP) pathway regulatory genes in Zingiber zerumbet following Pythium myriotylum infection.

• Metabolite analysis of in vitro plantlets treated with mevinolin (MEV) and fosmidomycin (FOS) reveals 54-fold decrease in zerumbone following FOS treatment.

• Identified plastidial IPP flux contributing more towards zerumbone biosynthesis.

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Nair, A.R., Divakaran, K. & Pillai, P. Evidence for Methylerythritol Pathway (MEP) Contributions to Zerumbone Biosynthesis as Revealed by Expression Analysis of Regulatory Genes and Metabolic Inhibitors Studies. Plant Mol Biol Rep 38, 370–379 (2020). https://doi.org/10.1007/s11105-020-01202-5

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