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Rice cytochrome P450 MAX1 homologs catalyze distinct steps in strigolactone biosynthesis

Nature Chemical Biology volume 10, pages 1028–1033 (2014)Cite this article

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Abstract

Strigolactones (SLs) are a class of phytohormones and rhizosphere signaling compounds with high structural diversity. Three enzymes, carotenoid isomerase DWARF27 and carotenoid cleavage dioxygenases CCD7 and CCD8, were previously shown to convert all-trans-β-carotene to carlactone (CL), the SL precursor. However, how CL is metabolized to SLs has remained elusive. Here, by reconstituting the SL biosynthetic pathway in Nicotiana benthamiana, we show that a rice homolog of Arabidopsis MORE AXILLARY GROWTH 1 (MAX1), encodes a cytochrome P450 CYP711 subfamily member that acts as a CL oxidase to stereoselectively convert CL into ent-2′-epi-5-deoxystrigol (B-C lactone ring formation), the presumed precursor of rice SLs. A protein encoded by a second rice MAX1 homolog then catalyzes the conversion of ent-2′-epi-5-deoxystrigol to orobanchol. We therefore report that two members of CYP711 enzymes can catalyze two distinct steps in SL biosynthesis, identifying the first enzymes involved in B-C ring closure and a subsequent structural diversification step of SLs.

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Figure 1: Current knowledge of the SL biosynthesis pathway.
Figure 2: Production of CL in N. benthamiana and its conversion to ent-2′-epi-5DS by OsMAX1s.
Figure 3: Production of orobanchol in N. benthamiana and yeast microsomes expressing Os1400.
Figure 4: Predicted docking of CL and ent-2′-epi-5DS in OsMAX1s.
Figure 5: Rice SL biosynthesis from (Z)-(R)-CL.

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Acknowledgements

We thank Y. Wang from the Institute of Genetics and Developmental Biology at the Chinese Academy of Science for the p35s:OsD27:PJTK13 plasmid and K. Yoneyama (Weed Science Center, Utsunomiya University, Utsunomiya, Japan) and T. Asami (Department of Applied Biological Chemistry, The University of Tokyo, Japan) for supplying SL standards. We thank J. Beekwilder and K. Cankar (Plant Research International, Wageningen, the Netherlands) for technical advice on the yeast assays and B. Ramakers (Nijmegen University) for technical support with the CD spectra measurement of CL. We thank A. Reeder from the Centre for Microscopy, Characterisation and Analysis (University of Western Australia (UWA)) and M. Clarke from the Centre for Metabolomics (UWA) for technical assistance and instrument access. We acknowledge funding by the Netherlands Organization for Scientific Research (VICI grant 865.06.002 and equipment grant 834.08.001 to H.J.B.), the Australian Research Council (LP0882775 for A.S. and FT110100304 for G.R.F.) and the UK Biotechnology and Biological Sciences Research Council (for J.H. and O.L.). Research reported in this publication was supported by the King Abdullah University of Science and Technology and was cofinanced by the Centre for BioSystems Genomics, which is part of the Netherlands Genomics Initiative/Netherlands Organization for Scientific Research.

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  1. Jo Hepworth

    Present address: Present address: John Innes Centre, Norwich Research Park, Norwich, UK.,

Authors and Affiliations

  1. Laboratory of Plant Physiology, Wageningen University, Wageningen, the Netherlands

    Yanxia Zhang, Tatsiana Charnikhova, Francel Verstappen, Sander van der Krol, Carolien Ruyter-Spira & Harro J Bouwmeester

  2. Applied Bioinformatics, Plant Research International, Wageningen, the Netherlands

    Aalt D J van Dijk

  3. School of Chemistry and Biochemistry, The University of Western Australia, Crawley, Australia

    Adrian Scaffidi, Gavin R Flematti & Steven M Smith

  4. Faculty of Biology, University of Freiburg, Freiburg, Germany

    Manuel Hofmann & Salim Al-Babili

  5. Department of Biology, University of York, Heslington, York, UK

    Jo Hepworth & Ottoline Leyser

  6. Sainsbury Laboratory, University of Cambridge, Cambridge, UK

    Ottoline Leyser

  7. Radboud University Nijmegen, Institute for Molecules and Materials, Cluster of Organic Chemistry, Nijmegen, the Netherlands

    Binne Zwanenburg

  8. BESE Division, Center for Desert Agriculture, King Abdullah University of Science and Technology (KAUST), Thuwal, Kingdom of Saudi Arabia

    Salim Al-Babili

  9. Bioscience, Plant Research International, Wageningen, the Netherlands

    Carolien Ruyter-Spira

  10. Centre for Biosystems Genomics, Wageningen, the Netherlands

    Harro J Bouwmeester

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Contributions

Y.Z., C.R.-S. and H.J.B. designed the research. Y.Z. performed the gene cloning, N. benthamiana agro-infiltration, yeast expression assay and part of the MRM-LC/MS analysis experiments; A.S. synthesized the standards for two CL isomers and four 5DS stereoisomers and did CD spectra analysis for all of the compounds; G.R.F. performed chirality analysis for all the samples and compounds; M.H. prepared the CL derived from the Escherichia coli expression assays; T.C. and F.V. developed the MRM-LC/MS analysis protocols and performed MRM-LC/MS analysis, and J.H. cloned rice chromosome 6 MAX1 Os5100. B.Z. synthesized the orobanchol standards. A.D.J.v.D. conducted the protein modeling and docking. Y.Z., A.D.J.v.D., A.S., G.R.F., S.v.d.K., O.L., S.M.S., B.Z., S.A.-B., C.R.-S. and H.J.B. were involved in data interpretation and writing of the manuscript. C.R.-S. and H.J.B. supervised the project. A.D.J.v.D., A.S., G.R.F. and M.H. contributed equally to this work. C.R.-S. and H.J.B. contributed equally to this work.

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Correspondence to Harro J Bouwmeester.

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Zhang, Y., van Dijk, A., Scaffidi, A. et al. Rice cytochrome P450 MAX1 homologs catalyze distinct steps in strigolactone biosynthesis. Nat Chem Biol 10, 1028–1033 (2014). https://doi.org/10.1038/nchembio.1660

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