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Title: Downregulation of a UDP-Arabinomutase Gene in Switchgrass (Panicum virgatum L.) Results in Increased Cell Wall Lignin While Reducing Arabinose-Glycans

Abstract

Switchgrass (Panicum virgatum L.) is a C 4 perennial prairie grass and a dedicated feedstock for lignocellulosic biofuels. Saccharification and biofuel yields are inhibited by the plant cell wall's natural recalcitrance against enzymatic degradation. Plant hemicellulose polysaccharides such as arabinoxylans structurally support and cross-link other cell wall polymers. Grasses predominately have Type II cell walls that are abundant in arabinoxylan, which comprise nearly 25% of aboveground biomass. A primary component of arabinoxylan synthesis is uridine diphosphate (UDP) linked to arabinofuranose (Araf). A family of UDP-arabinopyranose mutase (UAM)/reversible glycosylated polypeptides catalyze the interconversion between UDP-arabinopyranose (UDP-Arap) and UDP-Araf. The expression of a switchgrass arabinoxylan biosynthesis pathway gene, PvUAM1, was decreased via RNAi to investigate its role in cell wall recalcitrance in the feedstock. PvUAM1 encodes a switchgrass homolog of UDP-arabinose mutase, which converts UDP-Arap to UDP-Araf. Southern blot analysis revealed each transgenic line contained between one to at least seven T-DNA insertions, resulting in some cases, a 95% reduction of native PvUAM1 transcript in stem internodes. Transgenic plants had increased pigmentation in vascular tissues at nodes, but were otherwise similar in morphology to the non-transgenic control. Cell wall-associated arabinose was decreased in leaves and stems by over 50%, but there wasmore » an increase in cellulose. In addition, there was a commensurate change in arabinose side chain extension. Cell wall lignin composition was altered with a concurrent increase in lignin content and transcript abundance of lignin biosynthetic genes in mature tillers. Enzymatic saccharification efficiency was unchanged in the transgenic plants relative to the control. Plants with attenuated PvUAM1 transcript had increased cellulose and lignin in cell walls. A decrease in cell wall-associated arabinose was expected, which was likely caused by fewer Araf residues in the arabinoxylan. The decrease in arabinoxylan may cause a compensation response to maintain cell wall integrity by increasing cellulose and lignin biosynthesis. In cases in which increased lignin is desired, e.g., feedstocks for carbon fiber production, downregulated UAM1 coupled with altered expression of other arabinoxylan biosynthesis genes might result in even higher production of lignin in biomass.« less

Authors:
 [1];  [2];  [1];  [3];  [4];  [4];  [4];  [1];  [1];  [1];  [4];  [3];  [2];  [2];  [5];  [1]
  1. Univ. of Tennessee, Knoxville, TN (United States). Dept. of Plant Sciences; Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). BioEnergy Science Center (BESC)
  2. Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). BioEnergy Science Center (BESC); Univ. of Georgia, Athens, GA (United States). Complex Carbohydrate Research Center
  3. Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). BioEnergy Science Center (BESC); Samuel Roberts Noble Foundation, Ardmore, OK (United States)
  4. Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). BioEnergy Science Center (BESC); National Renewable Energy Lab. (NREL), Golden, CO (United States)
  5. Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). BioEnergy Science Center (BESC); Univ. of Georgia, Athens, GA (United States). Complex Carbohydrate Research Center; Univ. of Georgia, Athens, GA (United States). Plant Biology
Publication Date:
Research Org.:
National Renewable Energy Lab. (NREL), Golden, CO (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
OSTI Identifier:
1334742
Report Number(s):
NREL/JA-2700-67522
Journal ID: ISSN 1664-462X
Grant/Contract Number:  
AC36-08GO28308
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
Frontiers in Plant Science
Additional Journal Information:
Journal Volume: 7; Journal ID: ISSN 1664-462X
Publisher:
Frontiers Research Foundation
Country of Publication:
United States
Language:
English
Subject:
09 BIOMASS FUELS; switchgrass; hemicellulosearabinoxylan; UDP-arabinopyranosemutase/reversibleglycosylated polypeptide; biofuel; recalcitrance

Citation Formats

Willis, Jonathan D., Smith, James A., Mazarei, Mitra, Zhang, Ji-Yi, Turner, Geoffrey B., Decker, Stephen R., Sykes, Robert W., Poovaiah, Charleson R., Baxter, Holly L., Mann, David G. J., Davis, Mark F., Udvardi, Michael K., Peña, Maria J., Backe, Jason, Bar-Peled, Maor, and Stewart, C. N. Downregulation of a UDP-Arabinomutase Gene in Switchgrass (Panicum virgatum L.) Results in Increased Cell Wall Lignin While Reducing Arabinose-Glycans. United States: N. p., 2016. Web. doi:10.3389/fpls.2016.01580.
Willis, Jonathan D., Smith, James A., Mazarei, Mitra, Zhang, Ji-Yi, Turner, Geoffrey B., Decker, Stephen R., Sykes, Robert W., Poovaiah, Charleson R., Baxter, Holly L., Mann, David G. J., Davis, Mark F., Udvardi, Michael K., Peña, Maria J., Backe, Jason, Bar-Peled, Maor, & Stewart, C. N. Downregulation of a UDP-Arabinomutase Gene in Switchgrass (Panicum virgatum L.) Results in Increased Cell Wall Lignin While Reducing Arabinose-Glycans. United States. doi:10.3389/fpls.2016.01580.
Willis, Jonathan D., Smith, James A., Mazarei, Mitra, Zhang, Ji-Yi, Turner, Geoffrey B., Decker, Stephen R., Sykes, Robert W., Poovaiah, Charleson R., Baxter, Holly L., Mann, David G. J., Davis, Mark F., Udvardi, Michael K., Peña, Maria J., Backe, Jason, Bar-Peled, Maor, and Stewart, C. N. Wed . "Downregulation of a UDP-Arabinomutase Gene in Switchgrass (Panicum virgatum L.) Results in Increased Cell Wall Lignin While Reducing Arabinose-Glycans". United States. doi:10.3389/fpls.2016.01580. https://www.osti.gov/servlets/purl/1334742.
@article{osti_1334742,
title = {Downregulation of a UDP-Arabinomutase Gene in Switchgrass (Panicum virgatum L.) Results in Increased Cell Wall Lignin While Reducing Arabinose-Glycans},
author = {Willis, Jonathan D. and Smith, James A. and Mazarei, Mitra and Zhang, Ji-Yi and Turner, Geoffrey B. and Decker, Stephen R. and Sykes, Robert W. and Poovaiah, Charleson R. and Baxter, Holly L. and Mann, David G. J. and Davis, Mark F. and Udvardi, Michael K. and Peña, Maria J. and Backe, Jason and Bar-Peled, Maor and Stewart, C. N.},
abstractNote = {Switchgrass (Panicum virgatum L.) is a C4 perennial prairie grass and a dedicated feedstock for lignocellulosic biofuels. Saccharification and biofuel yields are inhibited by the plant cell wall's natural recalcitrance against enzymatic degradation. Plant hemicellulose polysaccharides such as arabinoxylans structurally support and cross-link other cell wall polymers. Grasses predominately have Type II cell walls that are abundant in arabinoxylan, which comprise nearly 25% of aboveground biomass. A primary component of arabinoxylan synthesis is uridine diphosphate (UDP) linked to arabinofuranose (Araf). A family of UDP-arabinopyranose mutase (UAM)/reversible glycosylated polypeptides catalyze the interconversion between UDP-arabinopyranose (UDP-Arap) and UDP-Araf. The expression of a switchgrass arabinoxylan biosynthesis pathway gene, PvUAM1, was decreased via RNAi to investigate its role in cell wall recalcitrance in the feedstock. PvUAM1 encodes a switchgrass homolog of UDP-arabinose mutase, which converts UDP-Arap to UDP-Araf. Southern blot analysis revealed each transgenic line contained between one to at least seven T-DNA insertions, resulting in some cases, a 95% reduction of native PvUAM1 transcript in stem internodes. Transgenic plants had increased pigmentation in vascular tissues at nodes, but were otherwise similar in morphology to the non-transgenic control. Cell wall-associated arabinose was decreased in leaves and stems by over 50%, but there was an increase in cellulose. In addition, there was a commensurate change in arabinose side chain extension. Cell wall lignin composition was altered with a concurrent increase in lignin content and transcript abundance of lignin biosynthetic genes in mature tillers. Enzymatic saccharification efficiency was unchanged in the transgenic plants relative to the control. Plants with attenuated PvUAM1 transcript had increased cellulose and lignin in cell walls. A decrease in cell wall-associated arabinose was expected, which was likely caused by fewer Araf residues in the arabinoxylan. The decrease in arabinoxylan may cause a compensation response to maintain cell wall integrity by increasing cellulose and lignin biosynthesis. In cases in which increased lignin is desired, e.g., feedstocks for carbon fiber production, downregulated UAM1 coupled with altered expression of other arabinoxylan biosynthesis genes might result in even higher production of lignin in biomass.},
doi = {10.3389/fpls.2016.01580},
journal = {Frontiers in Plant Science},
issn = {1664-462X},
number = ,
volume = 7,
place = {United States},
year = {2016},
month = {10}
}

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