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The subcellular localization of two isopentenyl diphosphate isomerases in rice suggests a role for the endoplasmic reticulum in isoprenoid biosynthesis

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Abstract

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Both OsIPPI1 and OsIPPI2 enzymes are found in the endoplasmic reticulum, providing novel important insights into the role of this compartment in the synthesis of MVA pathway isoprenoids.

Abstract

Isoprenoids are synthesized from the precursor’s isopentenyl diphosphate (IPP) and dimethylallyl diphosphosphate (DMAPP), which are interconverted by the enzyme isopentenyl diphosphate isomerase (IPPI). Many plants express multiple isoforms of IPPI, the only enzyme shared by the mevalonate (MVA) and non-mevalonate (MEP) pathways, but little is known about their specific roles. Rice (Oryza sativa) has two IPPI isoforms (OsIPPI1 and OsIPPI2). We, therefore, carried out a comprehensive comparison of IPPI gene expression, protein localization, and isoprenoid biosynthesis in this species. We found that OsIPPI1 mRNA was more abundant than OsIPPI2 mRNA in all tissues, and its expression in de-etiolated leaves mirrored the accumulation of phytosterols, suggesting a key role in the synthesis of MVA pathway isoprenoids. We investigated the subcellular localization of both isoforms by constitutively expressing them as fusions with synthetic green fluorescent protein. Both proteins localized to the endoplasmic reticulum (ER) as well as peroxisomes and mitochondria, whereas only OsIPPI2 was detected in plastids, due to an N-terminal transit peptide which is not present in OsIPPI1. Despite the plastidial location of OsIPPI2, the expression of OsIPPI2 mRNA did not mirror the accumulation of chlorophylls or carotenoids, indicating that OsIPPI2 may be a redundant component of the MEP pathway. The detection of both OsIPPI isoforms in the ER indicates that DMAPP can be synthesized de novo in this compartment. Our work shows that the ER plays an as yet unknown role in the synthesis of MVA-derived isoprenoids, with important implications for the metabolic engineering of isoprenoid biosynthesis in higher plants.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (31870278); the Spanish Ministry of Economy and Competitiveness (MINECO), Spain (RTI2018-097613-B-I00; PGC2018-097655-B-I00); and in part by the European Union Framework Program DISCO (613513) “from DISCOvery to products: a next-generation pipeline for the sustainable generation of high-value plant products”, the European Cooperation in Science and Technology project EUROCAROTEN (OC-2015-1-19780), Generalitat de Catalunya Grant 2017 SGR 828 to the Agricultural Biotechnology and Bioeconomy Unit (ABBU), and the International Science and Technology Cooperation Project 20190201013JC (from Jilin Provincial Science and Technology Department, China).

Funding

CZ conceived and designed the research. XJ, CB, LG, VM, MD, XN, YS, LS, TC, PDF, and CZ conducted the experiments. XJ, CB, LG, VM, MD, XN, YS, LS, TC, PDF, PC, and CZ analyzed the data. XJ, PC, and CZ wrote the manuscript. All authors read and approved the manuscript.

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Authors and Affiliations

  1. Department of Plant Production and Forestry Science, University of Lleida-Agrotecnio Center, Av. Alcalde Rovira Roure, 191, 25198, Lleida, Spain

    Xin Jin, Can Baysal, Vicente Medina, Teresa Capell, Paul Christou & Changfu Zhu

  2. School of Life Sciences, Changchun Normal University, Changchun, 130032, China

    Lihong Gao, Xiuzhen Ni, Yanmin Sheng & Changfu Zhu

  3. School of Life Sciences, Northeast Normal University, Changchun, 130024, China

    Lianxuan Shi

  4. School of Biological Sciences, Royal Holloway University of London, Egham Hill, Egham, Surrey, TW20 0EX, UK

    Margit Drapal & Paul D. Fraser

  5. ICREA, Catalan Institute for Research and Advanced Studies, Passeig Lluís Companys 23, 08010, Barcelona, Spain

    Paul Christou

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  1. Xin Jin
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Correspondence to Changfu Zhu.

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Jin, X., Baysal, C., Gao, L. et al. The subcellular localization of two isopentenyl diphosphate isomerases in rice suggests a role for the endoplasmic reticulum in isoprenoid biosynthesis. Plant Cell Rep 39, 119–133 (2020). https://doi.org/10.1007/s00299-019-02479-x

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