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Elucidation of gibberellin biosynthesis in bacteria reveals convergent evolution

Nature Chemical Biology volume 13, pages 69–74 (2017)Cite this article

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Abstract

Gibberellins (GAs) are crucial phytohormones involved in many aspects of plant growth and development, including plant–microbe interactions, which has led to GA production by plant-associated fungi and bacteria as well. While the GA biosynthetic pathways in plants and fungi have been elucidated and found to have arisen independently through convergent evolution, little has been uncovered about GA biosynthesis in bacteria. Some nitrogen-fixing, symbiotic, legume-associated rhizobia, including Bradyrhizobium japonicum—the symbiont of soybean—and Sinorhizobium fredii—a broad-host-nodulating species—contain a putative GA biosynthetic operon, or gene cluster. Through functional characterization of five unknown genes, we demonstrate that this operon encodes the enzymes necessary to produce GA9, thereby elucidating bacterial GA biosynthesis. The distinct nature of these enzymes indicates that bacteria have independently evolved a third biosynthetic pathway for GA production. Furthermore, our results also reveal a central biochemical logic that is followed in all three convergently evolved GA biosynthetic pathways.

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Figure 1: GA biosynthesis in rhizobia.
Figure 2: Summary of GA-biosynthetic-pathway reactions detected in incubations of B. japonicum gene-deletion bacteroids with GA precursors.
Figure 3: CYP117 is an ent-kaurene oxidase (KO).
Figure 4: CYP114 is an ent-kaurenoic acid oxidase (KAO) and requires FdGA for full functionality.
Figure 5: SDRGA oxidizes GA12-aldehyde (6) to produce GA12 (7).
Figure 6: CYP112 is a GA 20-oxidase.

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Acknowledgements

This work was supported by grants from the NIH (GM109773 to R.J.P.) and the Iowa Soybean Association (grant to R.J.P.); a Discovery Grant and an Engage Grant, both from Natural Sciences and Research Engineering Council of Canada (NSERC; grants to T.C.C.); an Ontario Graduate Scholarship (A.M.); the 20:20 Wheat Integrated Strategic Programme at Rothamsted Research, funded by the Biotechnology and Biological Sciences Research Council of the United Kingdom (support to P.H.); and the Fondo Nacional de Desarrollo Cientifico y Tecnologico (grant 1150797 to M.C.R.). Authentic ent-7α-hydroxykaurenoic acid (5), GA15 (8), and GA24 (9)-methyl ester were provided by P. Hedden. ent-[17-14C1]Kaurenoic acid, [17-14C1]GA12, [17-14C1]GA15, [17-14C1]GA24, and [17-14C1]GA9 were obtained from L. Mander (Australian National University, Canberra, Australia).

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Authors and Affiliations

  1. Roy J. Carver Department of Biochemistry, Biophysics, and Molecular Biology, Iowa State University, Ames, Iowa, USA

    Ryan S Nett, Xuan Lu, Raimund Nagel & Reuben J Peters

  2. Departamento de Química, Laboratorio de Bioorgánica, Facultad de Ciencias, Universidad de Chile, Santiago, Chile

    Mariana Montanares & Maria Cecilia Rojas

  3. Department of Biology, University of Waterloo, Waterloo, Ontario, Canada

    Ariana Marcassa & Trevor C Charles

  4. Rothamsted Research, Harpenden, Hertfordshire, UK

    Peter Hedden

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  1. Ryan S Nett
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Contributions

R.S.N. performed the heterologous expression assays, created many of the knockout bacterial strains, and wrote the manuscript. X.L. aided in cloning related to the heterologous expression assays and R.N. provided insightful data analysis. M.M. and M.C.R. performed the bacteroid incubation assays and HPLC analysis, and helped write the manuscript. A.M. and T.C.C. created several of the knockout bacterial strains. P.H. analyzed extracts from the bacteroid incubation assays by GC–MS and helped write the manuscript. R.J.P. conceived the project, aided in data analysis, and helped write the manuscript.

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Correspondence to Reuben J Peters.

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Nett, R., Montanares, M., Marcassa, A. et al. Elucidation of gibberellin biosynthesis in bacteria reveals convergent evolution. Nat Chem Biol 13, 69–74 (2017). https://doi.org/10.1038/nchembio.2232

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