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Role of bifunctional ammonia-lyase in grass cell wall biosynthesis

Nature Plants volume 2, Article number: 16050 (2016) Cite this article

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Abstract

L-Phenylalanine ammonia-lyase (PAL) is the first enzyme in the biosynthesis of phenylpropanoid-derived plant compounds such as flavonoids, coumarins and the cell wall polymer lignin. The cell walls of grasses possess higher proportions of syringyl (S)-rich lignins and high levels of esterified coumaric acid compared with those of dicotyledonous plants, and PAL from grasses can also possess tyrosine ammonia-lyase (TAL) activity, the reason for which has remained unclear. Using phylogenetic, transcriptomic and in vitro biochemical analyses, we identified a single homotetrameric bifunctional ammonia-lyase (PTAL) among eight BdPAL enzymes in the model grass species Brachypodium distachyon. 13C isotope labelling experiments along with BdPTAL1-downregulation in transgenic plants showed that the TAL activity of BdPTAL1 can provide nearly half of the total lignin deposited in Brachypodium, with a preference for S-lignin and wall-bound coumarate biosynthesis, indicating that PTAL function is linked to the characteristic features of grass cell walls. Furthermore, isotope dilution experiments suggest that the pathways to lignin from L-phenylalanine and L-tyrosine are distinct beyond the formation of 4-coumarate, supporting the organization of lignin synthesis enzymes in one or more metabolons.

Land colonization by plants in the Devonian was a key step towards the development of terrestrial ecosystems. Ancestral land plants had to face stresses such as ultraviolet irradiation, desiccation and attack by soil microbial communities, which drove key adaptations, including the emergence of specialized metabolic pathways1,2. L-Phenylalanine (L-Phe) and L-tyrosine (L-Tyr) are aromatic amino acids used for the synthesis of proteins, and in plants they also serve as precursors of numerous natural products with diverse biological functions such as signal molecules, cell wall structural components, phytoalexins, pigments and ultraviolet protectants3. Both these aromatic amino acids are derived from the shikimate pathway, to which more than 30% of photosynthetically fixed carbon is directed in vascular plants toward the synthesis of the lignin biopolymer via the phenylpropanopid pathway4.

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Figure 1: Phylogeny, gene expression and enzymatic activity of Brachypodium BdPALs.
Figure 2: Purification of the native PTAL enzyme from Brachypodium stem tissue.
Figure 3: Labelling of Brachypodium seedlings with 13C9-L-Phe and 13C9-L-Tyr.
Figure 4: Phenotype and lignin characteristics of Brachypodium PTAL1 RNAi lines.
Figure 5: Model for the early steps of lignin synthesis including both PAL and TAL pathways in monocot plants.

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Acknowledgements

We thank D. Huhman and the Noble Foundation Analytical Chemistry Facility for the analysis of soluble phenolic compounds, Dr X. Wang for assistance with protein purification, and Dr L. Gallego-Giraldo for critical reading of the manuscript. This work was supported by a Barrie Foundation Fellowship (to J.B.), The University of North Texas and the BioEnergy Science Center (Oak Ridge National Laboratory), a US Department of Energy (DOE) Bioenergy Research Center supported by the Office of Biological and Environmental Research in the DOE Office of Science.

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Authors and Affiliations

  1. BioDiscovery Institute, University of North Texas, Denton, 76203, Texas, USA

    Jaime Barros, Juan C. Serrani-Yarce, Fang Chen, David Baxter, Barney J. Venables & Richard A. Dixon

  2. Department of Biological Sciences, University of North Texas, Denton, 76203, Texas, USA

    Jaime Barros, Juan C. Serrani-Yarce, Fang Chen & Richard A. Dixon

  3. BioEnergy Science Center (BESC), Oak Ridge National Laboratory, Oak Ridge, 37831, Tennessee, USA

    Fang Chen & Richard A. Dixon

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  1. Jaime Barros
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Contributions

R.A.D. and J.B. conceived the study; J.B., J.S.-Y., F.C. and R.A.D. designed the experiments; J.B, J.S.-Y. and D.B. performed experimental work; J.B., J.S.-Y., D.B., B.V., F.C. and R.A.D. discussed and interpreted results; J.B. and R.A.D. wrote the paper.

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Correspondence to Richard A. Dixon.

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Barros, J., Serrani-Yarce, J., Chen, F. et al. Role of bifunctional ammonia-lyase in grass cell wall biosynthesis. Nature Plants 2, 16050 (2016). https://doi.org/10.1038/nplants.2016.50

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