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Title: Functional Analysis of Cellulose Synthase CesA4 and CesA6 Genes in Switchgrass ( Panicum virgatum) by Overexpression and RNAi-Mediated Gene Silencing

Abstract

Switchgrass (Panicum virgatum L.) is a leading lignocellulosic bioenergy feedstock. Cellulose is a major component of the plant cell walls and the primary substrate for saccharification. Accessibility of cellulose to enzymatic breakdown into fermentable sugars is limited by the presence of lignin in the plant cell wall. In this study, putatively novel switchgrass secondary cell wall cellulose synthase PvCesA4 and primary cell wall PvCesA6 genes were identified and their functional role in cellulose synthesis and cell wall composition was examined by overexpression and knockdown of the individual genes in switchgrass. The endogenous expression of PvCesA4 and PvCesA6 genes varied among including roots, leaves, stem, and reproductive tissues. Increasing or decreasing PvCesA4 and PvCesA6 expression to extreme levels in the transgenic lines resulted in decreased biomass production. PvCesA6-overexpressing lines had reduced lignin content and syringyl/guaiacyl lignin monomer ratio accompanied by increased sugar release efficiency, suggesting an impact of PvCesA6 expression levels on lignin biosynthesis. Cellulose content and cellulose crystallinity were decreased, while xylan content was increased in PvCesA4 and PvCesA6 overexpression or knockdown lines. The increase in xylan content suggests that the amount of non-cellulosic cell wall polysaccharide was modified in these plants. Taken together, the results show that the manipulationmore » of the cellulose synthase genes alters the cell wall composition and availability of cellulose as a bioprocessing substrate.« less

Authors:
 [1];  [1]; ORCiD logo [2];  [3];  [4];  [5]; ORCiD logo [2];  [1];  [6];  [7];  [7]; ORCiD logo [7];  [6];  [6];  [3];  [8];  [4];  [1]
  1. Univ. of Tennessee, Knoxville, TN (United States). Department of Plant Sciences; Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). BioEnergy Science Center
  2. Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). BioEnergy Science Center and Biosciences Division, Joint Institute for Biological Science
  3. Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). BioEnergy Science Center; Univ. of Georgia, Athens, GA (United States). Complex Carbohydrate Research Center
  4. Univ. of Tennessee, Knoxville, TN (United States). Center for Renewable Carbon
  5. Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). BioEnergy Science Center; Univ. of Tennessee, Knoxville, TN (United States). Department of Chemical and Biomolecular Engineering
  6. Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). BioEnergy Science Center; Noble Research Institute, Ardmore, OK (United States)
  7. National Renewable Energy Lab. (NREL), Golden, CO (United States)
  8. Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). BioEnergy Science Center and Biosciences Division, Joint Institute for Biological Science; Univ. of Tennessee, Knoxville, TN (United States). Department of Chemical and Biomolecular Engineering
Publication Date:
Research Org.:
National Renewable Energy Lab. (NREL), Golden, CO (United States); Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
OSTI Identifier:
1468520
Alternate Identifier(s):
OSTI ID: 1468056
Report Number(s):
[NREL/JA-2700-72304]
[Journal ID: ISSN 1664-462X]
Grant/Contract Number:  
[AC36-08GO28308; AC05-00OR22725]
Resource Type:
Accepted Manuscript
Journal Name:
Frontiers in Plant Science
Additional Journal Information:
[ Journal Volume: 9]; Journal ID: ISSN 1664-462X
Publisher:
Frontiers Research Foundation
Country of Publication:
United States
Language:
English
Subject:
09 BIOMASS FUELS; 59 BASIC BIOLOGICAL SCIENCES; biofuel; cellulose synthase; lignocellulosic; overexpression; PvCesA4; PvCesA6; RNAi-gene silencing; switchgrass

Citation Formats

Mazarei, Mitra, Baxter, Holly L., Li, Mi, Biswal, Ajaya K., Kim, Keonhee, Meng, Xianzhi, Pu, Yunqiao, Wuddineh, Wegi A., Zhang, Ji-Yi, Turner, Geoffrey B., Sykes, Robert W., Davis, Mark F., Udvardi, Michael K., Wang, Zeng-Yu, Mohnen, Debra, Ragauskas, Arthur J., Labbé, Nicole, and Stewart, C. Neal. Functional Analysis of Cellulose Synthase CesA4 and CesA6 Genes in Switchgrass (Panicum virgatum) by Overexpression and RNAi-Mediated Gene Silencing. United States: N. p., 2018. Web. doi:10.3389/fpls.2018.01114.
Mazarei, Mitra, Baxter, Holly L., Li, Mi, Biswal, Ajaya K., Kim, Keonhee, Meng, Xianzhi, Pu, Yunqiao, Wuddineh, Wegi A., Zhang, Ji-Yi, Turner, Geoffrey B., Sykes, Robert W., Davis, Mark F., Udvardi, Michael K., Wang, Zeng-Yu, Mohnen, Debra, Ragauskas, Arthur J., Labbé, Nicole, & Stewart, C. Neal. Functional Analysis of Cellulose Synthase CesA4 and CesA6 Genes in Switchgrass (Panicum virgatum) by Overexpression and RNAi-Mediated Gene Silencing. United States. doi:10.3389/fpls.2018.01114.
Mazarei, Mitra, Baxter, Holly L., Li, Mi, Biswal, Ajaya K., Kim, Keonhee, Meng, Xianzhi, Pu, Yunqiao, Wuddineh, Wegi A., Zhang, Ji-Yi, Turner, Geoffrey B., Sykes, Robert W., Davis, Mark F., Udvardi, Michael K., Wang, Zeng-Yu, Mohnen, Debra, Ragauskas, Arthur J., Labbé, Nicole, and Stewart, C. Neal. Fri . "Functional Analysis of Cellulose Synthase CesA4 and CesA6 Genes in Switchgrass (Panicum virgatum) by Overexpression and RNAi-Mediated Gene Silencing". United States. doi:10.3389/fpls.2018.01114. https://www.osti.gov/servlets/purl/1468520.
@article{osti_1468520,
title = {Functional Analysis of Cellulose Synthase CesA4 and CesA6 Genes in Switchgrass (Panicum virgatum) by Overexpression and RNAi-Mediated Gene Silencing},
author = {Mazarei, Mitra and Baxter, Holly L. and Li, Mi and Biswal, Ajaya K. and Kim, Keonhee and Meng, Xianzhi and Pu, Yunqiao and Wuddineh, Wegi A. and Zhang, Ji-Yi and Turner, Geoffrey B. and Sykes, Robert W. and Davis, Mark F. and Udvardi, Michael K. and Wang, Zeng-Yu and Mohnen, Debra and Ragauskas, Arthur J. and Labbé, Nicole and Stewart, C. Neal},
abstractNote = {Switchgrass (Panicum virgatum L.) is a leading lignocellulosic bioenergy feedstock. Cellulose is a major component of the plant cell walls and the primary substrate for saccharification. Accessibility of cellulose to enzymatic breakdown into fermentable sugars is limited by the presence of lignin in the plant cell wall. In this study, putatively novel switchgrass secondary cell wall cellulose synthase PvCesA4 and primary cell wall PvCesA6 genes were identified and their functional role in cellulose synthesis and cell wall composition was examined by overexpression and knockdown of the individual genes in switchgrass. The endogenous expression of PvCesA4 and PvCesA6 genes varied among including roots, leaves, stem, and reproductive tissues. Increasing or decreasing PvCesA4 and PvCesA6 expression to extreme levels in the transgenic lines resulted in decreased biomass production. PvCesA6-overexpressing lines had reduced lignin content and syringyl/guaiacyl lignin monomer ratio accompanied by increased sugar release efficiency, suggesting an impact of PvCesA6 expression levels on lignin biosynthesis. Cellulose content and cellulose crystallinity were decreased, while xylan content was increased in PvCesA4 and PvCesA6 overexpression or knockdown lines. The increase in xylan content suggests that the amount of non-cellulosic cell wall polysaccharide was modified in these plants. Taken together, the results show that the manipulation of the cellulose synthase genes alters the cell wall composition and availability of cellulose as a bioprocessing substrate.},
doi = {10.3389/fpls.2018.01114},
journal = {Frontiers in Plant Science},
number = ,
volume = [9],
place = {United States},
year = {2018},
month = {8}
}

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Works referenced in this record:

Functional analyses of cellulose synthase genes in flax ( Linum usitatissimum ) by virus-induced gene silencing
journal, February 2015

  • Chantreau, Maxime; Chabbert, Brigitte; Billiard, Sylvain
  • Plant Biotechnology Journal, Vol. 13, Issue 9
  • DOI: 10.1111/pbi.12350

Two-year field analysis of reduced recalcitrance transgenic switchgrass
journal, April 2014

  • Baxter, Holly L.; Mazarei, Mitra; Labbe, Nicole
  • Plant Biotechnology Journal, Vol. 12, Issue 7
  • DOI: 10.1111/pbi.12195

Lignin bioengineering
journal, April 2014


Sugar release and growth of biofuel crops are improved by downregulation of pectin biosynthesis
journal, February 2018

  • Biswal, Ajaya K.; Atmodjo, Melani A.; Li, Mi
  • Nature Biotechnology, Vol. 36, Issue 3
  • DOI: 10.1038/nbt.4067

Hemicelluloses
journal, June 2010


Cellulose microfibril crystallinity is reduced by mutating C-terminal transmembrane region residues CESA1A903V and CESA3T942I of cellulose synthase
journal, February 2012

  • Harris, D. M.; Corbin, K.; Wang, T.
  • Proceedings of the National Academy of Sciences, Vol. 109, Issue 11
  • DOI: 10.1073/pnas.1200352109

Genomics of cellulose biosynthesis in poplars: Research review
journal, August 2004


Lignocellulose conversion to biofuels: current challenges, global perspectives
journal, June 2009


Powerful regulatory systems and post-transcriptional gene silencing resist increases in cellulose content in cell walls of barley
journal, January 2015


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

Genetic loci simultaneously controlling lignin monomers and biomass digestibility of rice straw
journal, February 2018


Cell Expansion-Mediated Organ Growth Is Affected by Mutations in Three EXIGUA Genes
journal, May 2012


Growth of the plant cell wall
journal, November 2005

  • Cosgrove, Daniel J.
  • Nature Reviews Molecular Cell Biology, Vol. 6, Issue 11, p. 850-861
  • DOI: 10.1038/nrm1746

Interactions among three distinct CesA proteins essential for cellulose synthesis
journal, January 2003

  • Taylor, N. G.; Howells, R. M.; Huttly, A. K.
  • Proceedings of the National Academy of Sciences, Vol. 100, Issue 3
  • DOI: 10.1073/pnas.0337628100

Cellulose-Microtubule Uncoupling Proteins Prevent Lateral Displacement of Microtubules during Cellulose Synthesis in Arabidopsis
journal, August 2016


BdCESA7, BdCESA8, and BdPMT Utility Promoter Constructs for Targeted Expression to Secondary Cell-Wall-Forming Cells of Grasses
journal, February 2016

  • Petrik, Deborah L.; Cass, Cynthia L.; Padmakshan, Dharshana
  • Frontiers in Plant Science, Vol. 7
  • DOI: 10.3389/fpls.2016.00055

Hemicelluloses for fuel ethanol: A review
journal, July 2010


Expression of a Mutant Form of Cellulose Synthase AtCesA7 Causes Dominant Negative Effect on Cellulose Biosynthesis
journal, June 2003

  • Zhong, Ruiqin; Morrison, W. Herbert; Freshour, Glenn D.
  • Plant Physiology, Vol. 132, Issue 2
  • DOI: 10.1104/pp.102.019331

ORIGINAL RESEARCH: Lignocellulose recalcitrance screening by integrated high-throughput hydrothermal pretreatment and enzymatic saccharification
journal, April 2010

  • Selig, Michael J.; Tucker, Melvin P.; Sykes, Robert W.
  • Industrial Biotechnology, Vol. 6, Issue 2
  • DOI: 10.1089/ind.2010.0009

An unusual xylan in Arabidopsis primary cell walls is synthesised by GUX3, IRX9L, IRX10L and IRX14
journal, June 2015

  • Mortimer, Jenny C.; Faria-Blanc, Nuno; Yu, Xiaolan
  • The Plant Journal, Vol. 83, Issue 3
  • DOI: 10.1111/tpj.12898

Biomass Recalcitrance: Engineering Plants and Enzymes for Biofuels Production
journal, February 2007

  • Himmel, M. E.; Ding, S.-Y.; Johnson, D. K.
  • Science, Vol. 315, Issue 5813, p. 804-807
  • DOI: 10.1126/science.1137016

Cell wall mutants
journal, January 2000

  • Fagard, Mathilde; Höfte, Herman; Vernhettes, Samantha
  • Plant Physiology and Biochemistry, Vol. 38, Issue 1-2
  • DOI: 10.1016/S0981-9428(00)00171-6

Systems and synthetic biology approaches to alter plant cell walls and reduce biomass recalcitrance
journal, November 2014

  • Kalluri, Udaya C.; Yin, Hengfu; Yang, Xiaohan
  • Plant Biotechnology Journal, Vol. 12, Issue 9
  • DOI: 10.1111/pbi.12283

An update on the nomenclature for the cellulose synthase genes in Populus
journal, May 2009


Perturbation of Wood Cellulose Synthesis Causes Pleiotropic Effects in Transgenic Aspen
journal, March 2011

  • Joshi, Chandrashekhar P.; Thammannagowda, Shivegowda; Fujino, Takeshi
  • Molecular Plant, Vol. 4, Issue 2
  • DOI: 10.1093/mp/ssq081

Plant cellulose synthesis: CESA proteins crossing kingdoms
journal, April 2015


Cellulose Synthesis in Higher Plants
journal, November 2006


Three Distinct Rice Cellulose Synthase Catalytic Subunit Genes Required for Cellulose Synthesis in the Secondary Wall
journal, August 2003

  • Tanaka, Katsuyuki; Murata, Kazumasa; Yamazaki, Muneo
  • Plant Physiology, Vol. 133, Issue 1
  • DOI: 10.1104/pp.103.022442

Higher plants contain homologs of the bacterial celA genes encoding the catalytic subunit of cellulose synthase.
journal, October 1996

  • Pear, J. R.; Kawagoe, Y.; Schreckengost, W. E.
  • Proceedings of the National Academy of Sciences, Vol. 93, Issue 22
  • DOI: 10.1073/pnas.93.22.12637

Genetic evidence for three unique components in primary cell-wall cellulose synthase complexes in Arabidopsis
journal, September 2007

  • Persson, S.; Paredez, A.; Carroll, A.
  • Proceedings of the National Academy of Sciences, Vol. 104, Issue 39
  • DOI: 10.1073/pnas.0706592104

Development of an integrated transcript sequence database and a gene expression atlas for gene discovery and analysis in switchgrass ( Panicum virgatum L.)
journal, February 2013

  • Zhang, Ji-Yi; Lee, Yi-Ching; Torres-Jerez, Ivone
  • The Plant Journal, Vol. 74, Issue 1
  • DOI: 10.1111/tpj.12104

Toward a Systems Approach to Understanding Plant Cell Walls
journal, December 2004


Expression profiling and integrative analysis of the CESA/CSL superfamily in rice
journal, January 2010


Study of traits and recalcitrance reduction of field-grown COMT down-regulated switchgrass
journal, January 2017


Three AtCesA6-like members enhance biomass production by distinctively promoting cell growth in Arabidopsis
journal, October 2017

  • Hu, Huizhen; Zhang, Ran; Feng, Shengqiu
  • Plant Biotechnology Journal, Vol. 16, Issue 5
  • DOI: 10.1111/pbi.12842

Virus-Induced Silencing of a Plant Cellulose Synthase Gene
journal, May 2000

  • Burton, Rachel A.; Gibeaut, David M.; Bacic, Antony
  • The Plant Cell, Vol. 12, Issue 5
  • DOI: 10.1105/tpc.12.5.691

Cell-wall structure and anisotropy in procuste, a cellulose synthase mutant of Arabidopsis thaliana
journal, January 2006


Cellulose synthesis in maize: isolation and expression analysis of the cellulose synthase (CesA) gene family
journal, September 2004


Perturbation of Brachypodium distachyon CELLULOSE SYNTHASE A4 or 7 results in abnormal cell walls
journal, January 2013

  • Handakumbura, Pubudu P.; Matos, Dominick A.; Osmont, Karen S.
  • BMC Plant Biology, Vol. 13, Issue 1
  • DOI: 10.1186/1471-2229-13-131

Plant cell wall reconstruction toward improved lignocellulosic production and processability
journal, February 2010


A missense mutation in the transmembrane domain of CESA9 affects cell wall biosynthesis and plant growth in rice
journal, November 2012


Developmental Control of Lignification in Stems of Lowland Switchgrass Variety Alamo and the Effects on Saccharification Efficiency
journal, October 2009


Cellulose Synthases and Synthesis in Arabidopsis
journal, March 2011

  • Endler, Anne; Persson, Staffan
  • Molecular Plant, Vol. 4, Issue 2
  • DOI: 10.1093/mp/ssq079

Four alleles of AtCESA3 form an allelic series with respect to root phenotype in Arabidopsis thaliana
journal, March 2012


Disruption of Secondary Wall Cellulose Biosynthesis Alters Cadmium Translocation and Tolerance in Rice Plants
journal, May 2013

  • Song, Xue-Qin; Liu, Li-Feng; Jiang, Yi-Jun
  • Molecular Plant, Vol. 6, Issue 3
  • DOI: 10.1093/mp/sst025

Genetic Evidence That Cellulose Synthase Activity Influences Microtubule Cortical Array Organization
journal, June 2008

  • Paredez, Alexander R.; Persson, Staffan; Ehrhardt, David W.
  • Plant Physiology, Vol. 147, Issue 4
  • DOI: 10.1104/pp.108.120196

Describing and Quantifying Growth Stages of Perennial Forage Grasses
journal, January 1991


Characterization of cellulose structure of Populus plants modified in candidate cellulose biosynthesis genes
journal, November 2016


PROCUSTE1 Encodes a Cellulose Synthase Required for Normal Cell Elongation Specifically in Roots and Dark-Grown Hypocotyls of Arabidopsis
journal, December 2000

  • Fagard, Mathilde; Desnos, Thierry; Desprez, Thierry
  • The Plant Cell, Vol. 12, Issue 12
  • DOI: 10.1105/tpc.12.12.2409

Organization of cellulose synthase complexes involved in primary cell wall synthesis in Arabidopsis thaliana
journal, September 2007

  • Desprez, T.; Juraniec, M.; Crowell, E. F.
  • Proceedings of the National Academy of Sciences, Vol. 104, Issue 39
  • DOI: 10.1073/pnas.0706569104

Standardization of Switchgrass Sample Collection for Cell Wall and Biomass Trait Analysis
journal, January 2013


Genetic modification of plant cell walls to enhance biomass yield and biofuel production in bioenergy crops
journal, September 2016


Dynamic changes in transcriptome and cell wall composition underlying brassinosteroid-mediated lignification of switchgrass suspension cells
journal, November 2017


The Arabidopsis Mutant cev1 Links Cell Wall Signaling to Jasmonate and Ethylene Responses
journal, July 2002

  • Ellis, Christine; Karafyllidis, Ioannis; Wasternack, Claus
  • The Plant Cell, Vol. 14, Issue 7
  • DOI: 10.1105/tpc.002022

Rice Brittle culm 6 encodes a dominant-negative form of CesA protein that perturbs cellulose synthesis in secondary cell walls
journal, January 2011

  • Kotake, Toshihisa; Aohara, Tsutomu; Hirano, Ko
  • Journal of Experimental Botany, Vol. 62, Issue 6
  • DOI: 10.1093/jxb/erq395

Cellulose and callose synthesis and organization in focus, what's new?
journal, December 2016


The genome sequence of black cottonwood (Populus trichocarpa) reveals 18 conserved cellulose synthase (CesA) genes
journal, June 2005


Agrobacterium-Mediated Transformation of Switchgrass and Inheritance of the Transgenes
journal, October 2009


Field Evaluation of Transgenic Switchgrass Plants Overexpressing PvMYB4 for Reduced Biomass Recalcitrance
journal, January 2015

  • Baxter, Holly L.; Poovaiah, Charleson R.; Yee, Kelsey L.
  • BioEnergy Research, Vol. 8, Issue 3
  • DOI: 10.1007/s12155-014-9570-1

The CesA Gene Family of Barley. Quantitative Analysis of Transcripts Reveals Two Groups of Co-Expressed Genes
journal, December 2003

  • Burton, Rachel A.; Shirley, Neil J.; King, Brendon J.
  • Plant Physiology, Vol. 134, Issue 1
  • DOI: 10.1104/pp.103.032904

Monolignol Ferulate Transferase Introduces Chemically Labile Linkages into the Lignin Backbone
journal, April 2014


Disruption of Mediator rescues the stunted growth of a lignin-deficient Arabidopsis mutant
journal, March 2014

  • Bonawitz, Nicholas D.; Kim, Jeong Im; Tobimatsu, Yuki
  • Nature, Vol. 509, Issue 7500, p. 376-380
  • DOI: 10.1038/nature13084

Cellulose factories: advancing bioenergy production from forest trees
journal, November 2011


Engineering of a high-throughput screening system to identify cellulosic biomass, pretreatments, and enzyme formulations that enhance sugar release
journal, February 2010

  • Studer, Michael H.; DeMartini, Jaclyn D.; Brethauer, Simone
  • Biotechnology and Bioengineering, Vol. 105, Issue 2, p. 231-238
  • DOI: 10.1002/bit.22527

MEGA6: Molecular Evolutionary Genetics Analysis Version 6.0
journal, October 2013

  • Tamura, Koichiro; Stecher, Glen; Peterson, Daniel
  • Molecular Biology and Evolution, Vol. 30, Issue 12
  • DOI: 10.1093/molbev/mst197

Plant cell wall engineering: applications in biofuel production and improved human health
journal, April 2014