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Title: Downregulation of p-coumaroyl quinate/shikimate 3'-hydroxylase (C3'H) or cinnamate-4-hydrolylase (C4H) in Eucalyptus urophylla x Eucalyptus grandis leads to increased extractability

Abstract

Lignin reduction through breeding and genetic modification has the potential to reduce costs in biomass processing in pulp and paper, forage, and lignocellulosic ethanol industries. Here, we present detailed characterization of the extractability and lignin structure of Eucalyptus urophylla x Eucalyptus grandis RNAi downregulated in p-coumaroyl quinate/shikimate 3'-hydroxylase (C3'H) or cinnamate-4-hydroxylase (C4H). Both the C3'H and C4H downregulated lines were found to have significantly higher extractability when exposed to NaOH base extraction, indicating altered cell wall construction. The molecular weight of isolated lignin was measured and lignin structure was determined by HSQC NMR-based lignin subunit analysis for control and the C3'H and C4H downregulated lines. The slight reductions in average molecular weights of the lignin isolated from the transgenic lines (C3'H = 7000, C4H = 6500, control = 7300) does not appear to explain the difference in extractability. The HSQC NMR-based lignin subunit analysis showed increases in H lignin content for the C3'H but only slight differences in the lignin subunit structure of the C3'H and C4H downregulated lines when compared to the control. The greatest difference between the C3'H and C4H downregulated lines is the total lignin content; therefore, it appears that overall lowered lignin content contributes greatly tomore » reduced recalcitrance and increased extractability of cell wall biopolymers. Furthermore, studies will be conducted to determine how the reduction in lignin content creates a less rigid cell wall that is more prone to extraction and sugar release.« less

Authors:
 [1];  [1];  [2];  [1];  [1];  [1];  [1]
  1. National Renewable Energy Lab. (NREL), Golden, CO (United States)
  2. ArborGen Inc., Ridgeville, SC (United States)
Publication Date:
Research Org.:
National Renewable Energy Lab. (NREL), Golden, CO (United States)
Sponsoring Org.:
USDOE Office of Science (SC)
OSTI Identifier:
1503054
Alternate Identifier(s):
OSTI ID: 1253985
Report Number(s):
NREL/JA-5100-65768
Journal ID: ISSN 1939-1234
Grant/Contract Number:  
AC36-08GO28308; BioEnergy Scence Center
Resource Type:
Published Article
Journal Name:
BioEnergy Research
Additional Journal Information:
Journal Volume: 9; Journal Issue: 2; Related Information: BioEnergy Research; Journal ID: ISSN 1939-1234
Publisher:
Springer
Country of Publication:
United States
Language:
English
Subject:
37 INORGANIC, ORGANIC, PHYSICAL, AND ANAYLYTICAL CHEMISTRY; nuclear magnetic resonance (NMR); plant; cell wall; transgenic; spectroscopy; eucalyptus; C3'H; C4H; lignin

Citation Formats

Ziebell, Angela, Gjersing, Erica, Hinchee, Maud, Katahira, Rui, Sykes, Robert W., Johnson, David K., and Davis, Mark F. Downregulation of p-coumaroyl quinate/shikimate 3'-hydroxylase (C3'H) or cinnamate-4-hydrolylase (C4H) in Eucalyptus urophylla x Eucalyptus grandis leads to increased extractability. United States: N. p., 2016. Web. doi:10.1007/s12155-016-9713-7.
Ziebell, Angela, Gjersing, Erica, Hinchee, Maud, Katahira, Rui, Sykes, Robert W., Johnson, David K., & Davis, Mark F. Downregulation of p-coumaroyl quinate/shikimate 3'-hydroxylase (C3'H) or cinnamate-4-hydrolylase (C4H) in Eucalyptus urophylla x Eucalyptus grandis leads to increased extractability. United States. doi:10.1007/s12155-016-9713-7.
Ziebell, Angela, Gjersing, Erica, Hinchee, Maud, Katahira, Rui, Sykes, Robert W., Johnson, David K., and Davis, Mark F. Wed . "Downregulation of p-coumaroyl quinate/shikimate 3'-hydroxylase (C3'H) or cinnamate-4-hydrolylase (C4H) in Eucalyptus urophylla x Eucalyptus grandis leads to increased extractability". United States. doi:10.1007/s12155-016-9713-7.
@article{osti_1503054,
title = {Downregulation of p-coumaroyl quinate/shikimate 3'-hydroxylase (C3'H) or cinnamate-4-hydrolylase (C4H) in Eucalyptus urophylla x Eucalyptus grandis leads to increased extractability},
author = {Ziebell, Angela and Gjersing, Erica and Hinchee, Maud and Katahira, Rui and Sykes, Robert W. and Johnson, David K. and Davis, Mark F.},
abstractNote = {Lignin reduction through breeding and genetic modification has the potential to reduce costs in biomass processing in pulp and paper, forage, and lignocellulosic ethanol industries. Here, we present detailed characterization of the extractability and lignin structure of Eucalyptus urophylla x Eucalyptus grandis RNAi downregulated in p-coumaroyl quinate/shikimate 3'-hydroxylase (C3'H) or cinnamate-4-hydroxylase (C4H). Both the C3'H and C4H downregulated lines were found to have significantly higher extractability when exposed to NaOH base extraction, indicating altered cell wall construction. The molecular weight of isolated lignin was measured and lignin structure was determined by HSQC NMR-based lignin subunit analysis for control and the C3'H and C4H downregulated lines. The slight reductions in average molecular weights of the lignin isolated from the transgenic lines (C3'H = 7000, C4H = 6500, control = 7300) does not appear to explain the difference in extractability. The HSQC NMR-based lignin subunit analysis showed increases in H lignin content for the C3'H but only slight differences in the lignin subunit structure of the C3'H and C4H downregulated lines when compared to the control. The greatest difference between the C3'H and C4H downregulated lines is the total lignin content; therefore, it appears that overall lowered lignin content contributes greatly to reduced recalcitrance and increased extractability of cell wall biopolymers. Furthermore, studies will be conducted to determine how the reduction in lignin content creates a less rigid cell wall that is more prone to extraction and sugar release.},
doi = {10.1007/s12155-016-9713-7},
journal = {BioEnergy Research},
number = 2,
volume = 9,
place = {United States},
year = {2016},
month = {1}
}

Journal Article:
Free Publicly Available Full Text
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DOI: 10.1007/s12155-016-9713-7

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Works referenced in this record:

Cell wall modifications triggered by the down-regulation of Coumarate 3-hydroxylase-1 in maize
journal, July 2015


A CP/MAS 13C-NMR study of cellulose structure on the surface of refined kraft pulp fibers
journal, August 2002


Radical Coupling Reactions in Lignin Synthesis: A Density Functional Theory Study
journal, April 2012

  • Sangha, Amandeep K.; Parks, Jerry M.; Standaert, Robert F.
  • The Journal of Physical Chemistry B, Vol. 116, Issue 16
  • DOI: 10.1021/jp2122449

Phenolic acid bridges between polysaccharides and lignin in wheat internodes
journal, January 1990


Structural Analysis of Wheat Straw Lignin by Quantitative 31 P and 2D NMR Spectroscopy. The Occurrence of Ester Bonds and α-O-4 Substructures
journal, April 1997

  • Crestini, Claudia; Argyropoulos, Dimitris S.
  • Journal of Agricultural and Food Chemistry, Vol. 45, Issue 4
  • DOI: 10.1021/jf960568k

RNAi-mediated suppression of p-coumaroyl-CoA 3'-hydroxylase in hybrid poplar impacts lignin deposition and soluble secondary metabolism
journal, March 2008

  • Coleman, H. D.; Park, J. -Y.; Nair, R.
  • Proceedings of the National Academy of Sciences, Vol. 105, Issue 11
  • DOI: 10.1073/pnas.0706537105

The Arabidopsis REF8 gene encodes the 3-hydroxylase of phenylpropanoid metabolism
journal, April 2002


Lignin modification improves fermentable sugar yields for biofuel production
journal, June 2007

  • Chen, Fang; Dixon, Richard A.
  • Nature Biotechnology, Vol. 25, Issue 7, p. 759-761
  • DOI: 10.1038/nbt1316

Changes in secondary metabolism and deposition of an unusual lignin in the ref8 mutant of Arabidopsis
journal, April 2002


NMR Characterization of C3H and HCT Down-Regulated Alfalfa Lignin
journal, October 2009


Comparative Studies on Cellulolytic Enzyme Lignin and Milled Wood Lignin of Sweetgum and Spruce
journal, January 1975


Whole plant cell wall characterization using solution-state 2D NMR
journal, August 2012


Exploring lignification in conifers by silencing hydroxycinnamoyl-CoA:shikimate hydroxycinnamoyltransferase in Pinus radiata
journal, July 2007

  • Wagner, A.; Ralph, J.; Akiyama, T.
  • Proceedings of the National Academy of Sciences, Vol. 104, Issue 28
  • DOI: 10.1073/pnas.0701428104

Designed for deconstruction - poplar trees altered in cell wall lignification improve the efficacy of bioethanol production
journal, January 2012


Cinnamic acid bridges between cell wall polymers in wheat and phalaris internodes
journal, April 1992


Lignin in straw of herbaceous crops
journal, November 2008


Enhanced characteristics of genetically modified switchgrass (Panicum virgatum L.) for high biofuel production
journal, January 2013

  • Shen, Hui; Poovaiah, Charleson R.; Ziebell, Angela
  • Biotechnology for Biofuels, Vol. 6, Issue 1
  • DOI: 10.1186/1754-6834-6-71

Recent Advances in Understanding Lignin Biosynthesis
journal, June 1998

  • Whetten, Ross W.; MacKay, John J.; Sederoff, Ronald R.
  • Annual Review of Plant Physiology and Plant Molecular Biology, Vol. 49, Issue 1
  • DOI: 10.1146/annurev.arplant.49.1.585

Effects of Coumarate 3-Hydroxylase Down-regulation on Lignin Structure
journal, January 2006

  • Ralph, John; Akiyama, Takuya; Kim, Hoon
  • Journal of Biological Chemistry, Vol. 281, Issue 13, p. 8843-8853
  • DOI: 10.1074/jbc.M511598200

Solid-State 13 C NMR Determination of the Syringyl/Guaiacyl Ratio in Hardwoods
journal, January 1987


Targeted down-regulation of cytochrome P450 enzymes for forage quality improvement in alfalfa (Medicago sativa L.)
journal, November 2005

  • Reddy, M. S. S.; Chen, F.; Shadle, G.
  • Proceedings of the National Academy of Sciences, Vol. 102, Issue 46
  • DOI: 10.1073/pnas.0505749102

Crystallinity of cellulose, as determined by CP/MAS NMR and XRD methods
journal, March 1987

  • Tee��r, R.; Serimaa, R.; Paakkarl, T.
  • Polymer Bulletin, Vol. 17, Issue 3
  • DOI: 10.1007/BF00285355

The Genetics of Lignin Biosynthesis: Connecting Genotype to Phenotype
journal, December 2010


Increase in 4-Coumaryl Alcohol Units during Lignification in Alfalfa ( Medicago sativa ) Alters the Extractability and Molecular Weight of Lignin
journal, October 2010

  • Ziebell, Angela; Gracom, Kristen; Katahira, Rui
  • Journal of Biological Chemistry, Vol. 285, Issue 50
  • DOI: 10.1074/jbc.M110.137315

Selective lignin downregulation leads to constitutive defense response expression in alfalfa (Medicago sativa L.)
journal, January 2011


Monolignol biosynthesis and genetic engineering of lignin in trees, a review
journal, July 2006


Pathway of p-Coumaric Acid Incorporation into Maize Lignin As Revealed by NMR
journal, October 1994

  • Ralph, John; Hatfield, Ronald D.; Quideau, Stephane
  • Journal of the American Chemical Society, Vol. 116, Issue 21, p. 9448-9456
  • DOI: 10.1021/ja00100a006

Solution-state 2D NMR of Ball-milled Plant Cell Wall Gels in DMSO-d 6
journal, March 2008


An Unusual Lignin from Kenaf
journal, January 1996

  • Ralph, John
  • Journal of Natural Products, Vol. 59, Issue 4, p. 341-342
  • DOI: 10.1021/np960143s

Rapid and simple determination of O-acetyl groups bound to plant cell walls by acid hydrolysis and 1H NMR measurement
journal, March 1994


Measuring the crystallinity index of cellulose by solid state 13C nuclear magnetic resonance
journal, June 2009