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Proteome Response to Arogenate Dehydratase Changes in Arabidopsis and Poplar


EMSL Project ID
48042

Abstract

Arogenate dehydratases (ADTs) are a class of isoenzymes involved in the production of phenylalanine (Phe). In addition to its involvement in protein biosynthesis, Phe is the starting metabolite in the phenylpropanoid pathway in plant systems. Designing plant biomass with specific control of phenylpropanoid metabolite production is of great interest right now, as there are a plethora of high value biofuel/biochemicals (e.g., styrenes, aroma/flavor compounds, 2-phenylethanol) that can be derived via these pathways. Lignin too is synthesized in great quantities by the phenylpropanoid pathway, and by designing biomass producing plants to limit or control lignin deposition, one could more effectively extract the cell wall sugar polymers used for the production of bioethanol. Controlling lignin deposition in plant biomass as well as designing plants to produce biochemicals/fuels of interest fits with the mission of EMSL and the DOE to develop sustainable sources of both biofuels and biochemical in order to aid the US in reducing its dependence on foreign and domestic petroleum supplies. We have successfully constructed numerous ADT knockout (KO) mutants (in Arabidopsis) and RNAi silencing mutants (in Poplar), as well as ADT overexpression lines in both organisms. Initial studies of these ADT mutants (GUS staining, phenotypic evaluation, lignin staining coupled with microscopy, monolignol measurements) have shown that alterations in the ADT composition in both plants produce profound temporal and spatial effects especially in regards to lignin deposition. The proteome analysis proposed here will complement our multi-omic approach to modeling the role of the individual ADTs. Through the study of all proposed knockout and overexpression lines, we will have a better understanding of how to control carbon flux into or away from the phenylpropanoid pathway and what if any positive or negative impacts, the compositional ADT changes have on the other pathways, and on metabolite production. By integrating all information captured from our multiple omics datasets we will be able to identify not only protein responses but also potential post-transcriptional regulation points, as these pathway points of interest are often identifiable when proteome and transcript data diverge from expected positive correlations in abundance. Taken together, these data will help us form a more accurate systems biology guided model to aid in future strategies to create designer biomass plants capable of producing multiple biofuels/biochemical.

Project Details

Project type
Exploratory Research
Start Date
2013-12-02
End Date
2014-09-30
Status
Closed

Team

Principal Investigator

Norman Lewis
Institution
Washington State University

Team Members

Kim Hixson
Institution
Pacific Northwest National Laboratory

Related Publications

Callum J. Bell, Joseph Brown, Therese R. Clauss, Laurence B. Davin, Kim K. Hixson, Norman G. Lewis, Mary S. Lipton, Joaquim V. Marques, Jason E. McDermott, Matthew E. Monroe, Ronald J. Moore, Ljiljana Paša-Tolić, Karl K. Weitz, Jason P. Wendler. 2021. "New Insights Into Lignification via Network and Multi-Omics Analyses of Arogenate Dehydratase Knock-Out Mutants in Arabidopsis thaliana." Frontiers in Plant Science 12 10.3389/fpls.2021.664250
Hohner R., J.V. Marques, T. Ito, Y. Amakura, A.D. Budgeon, K.K. Weitz, and K.K. Hixson, et al. 2018. "Reduced Arogenate Dehydratase Expression: Ramifications for Photosynthesis and Metabolism." Plant Physiology 177, no. 1:115-131. PNNL-SA-131412. doi:10.1104/pp.17.01766