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Modulation of auxin formation by the cytosolic phenylalanine biosynthetic pathway

Nature Chemical Biology volume 16, pages 850–856 (2020)Cite this article

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Abstract

In plants, phenylalanine biosynthesis occurs via two compartmentally separated pathways. Overexpression of petunia chorismate mutase 2 (PhCM2), which catalyzes the committed step of the cytosolic pathway, increased flux in cytosolic phenylalanine biosynthesis, but paradoxically decreased the overall levels of phenylalanine and phenylalanine-derived volatiles. Concomitantly, the levels of auxins, including indole-3-acetic acid and its precursor indole-3-pyruvic acid, were elevated. Biochemical and genetic analyses revealed the existence of metabolic crosstalk between the cytosolic phenylalanine biosynthesis and tryptophan-dependent auxin biosynthesis mediated by an aminotransferase that uses a cytosolic phenylalanine biosynthetic pathway intermediate, phenylpyruvate, as an amino acceptor for auxin formation.

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Fig. 1: Metabolic effects of PhCM2 overexpression in petunia flowers.
Fig. 2: Effect of application of different exogenous Trp-AT amino acceptors on auxin production.
Fig. 3: Metabolic effects of AtVAS1 and PhCM2 overexpression in petunia flowers.
Fig. 4: Proposed metabolic cross-talk between cytosolic phenylalanine biosynthetic pathway and auxin formation.

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Data availability

All figures except Fig. 4 have associated raw data available from the corresponding author upon reasonable request. Plant material generated in this study is available from the corresponding author upon request. The PhTrp-AT sequence reported in this paper has been deposited in GenBank database under accession number MN233649.

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Acknowledgements

This work was supported by grant MCB-1519083 from the US National Science Foundation to J.A.M. and N.D., by the USDA National Institute of Food and Agriculture Hatch project 177845 to N.D., and by the USDA National Institute of Food and Agriculture postdoctoral grant 2016-67012-24699 to J.H.L. The authors thank A. Withrow (Michigan State University) for assisting with transmission electron microscopy and B. Dilkes for discussion of Arabidopsis experiments with gravitropism.

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Author notes

  1. Yichun Qian

    Present address: Center for Genomics and Systems Biology, Department of Biology, New York University, New York, NY, USA

  2. Longyun Guo

    Present address: WuXi Biologics, Wuxi, China

  3. Alekzander S. Garcia

    Present address: Department of Chemistry, University of Illinois at Urbana-Champaign, Urbana, IL, USA

  4. These authors contributed equally: Joseph H. Lynch, Yichun Qian.

Authors and Affiliations

  1. Department of Biochemistry, Purdue University, West Lafayette, IN, USA

    Joseph H. Lynch, Longyun Guo, Itay Maoz, Xing-Qi Huang, Alekzander S. Garcia, John A. Morgan & Natalia Dudareva

  2. Department of Horticulture and Landscape Architecture, Purdue University, West Lafayette, IN, USA

    Yichun Qian & Natalia Dudareva

  3. Jack H Skirball Center for Chemical Biology and Proteomics, Salk Institute for Biological Studies, La Jolla, CA, USA

    Gordon Louie, Marianne E. Bowman & Joseph P. Noel

  4. Hughes Medical Institute, Salk Institute for Biological Studies, La Jolla, CA, USA

    Joseph P. Noel

  5. Davidson School of Chemical Engineering, Purdue University, West Lafayette, IN, USA

    John A. Morgan

  6. Purdue Center for Plant Biology, Purdue University, West Lafayette, IN, USA

    Natalia Dudareva

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Contributions

J.H.L., Y.Q., and N.D. designed research; Y.Q. generated PhCM2- and AtVAS-overexpression transgenic plants, and performed their metabolic profiling, analysis of chorismate mutase activity, and stable isotope labeling experiments. J.H.L. performed starch and auxin profiling of petunia transgenic plants and Arabidopsis cm2 mutants, feeding experiments with IAA and Trp-AT amino acceptors, and biochemical characterization of PhTrp-AT, and prepared samples for transmission electron microscopy. A.G. performed starch and auxin profiling in petunia petals. L.G. performed metabolic flux modeling. I.M. analyzed PhTrp-AT expression. X.-Q.H. analyzed PhTrp-At subcellular localization. G.L. and M.E.B. contributed to biochemical characterization of aminotransferase. Y.Q., J.H.L., L.G., I.M., X-Q.H., G.L., M.E.B., J.P.N., J.A.M., and N.D. analyzed data. J.H.L. and N.D. wrote the paper. All authors read and edited the manuscript.

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Correspondence to Natalia Dudareva.

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Lynch, J.H., Qian, Y., Guo, L. et al. Modulation of auxin formation by the cytosolic phenylalanine biosynthetic pathway. Nat Chem Biol 16, 850–856 (2020). https://doi.org/10.1038/s41589-020-0519-8

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