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Title: Identification and overexpression of a knotted1-like transcription factor in switchgrass ( Panicum virgatum L.) for lignocellulosic feedstock improvement

Abstract

High biomass production and wide adaptation has made switchgrass ( Panicum virgatum L.) an important candidate lignocellulosic bioenergy crop. One major limitation of this and other lignocellulosic feedstocks is the recalcitrance of complex carbohydrates to hydrolysis for conversion to biofuels. Lignin is the major contributor to recalcitrance as it limits the accessibility of cell wall carbohydrates to enzymatic breakdown into fermentable sugars. Therefore, genetic manipulation of the lignin biosynthesis pathway is one strategy to reduce recalcitrance. Here, we identified a switchgrass Knotted1 transcription factor, PvKN1, with the aim of genetically engineering switchgrass for reduced biomass recalcitrance for biofuel production. Gene expression of the endogenous PvKN1 gene was observed to be highest in young inflorescences and stems. Ectopic overexpression of PvKN1 in switchgrass altered growth, especially in early developmental stages. Transgenic lines had reduced expression of most lignin biosynthetic genes accompanied by a reduction in lignin content suggesting the involvement of PvKN1 in the broad regulation of the lignin biosynthesis pathway. Moreover, the reduced expression of the Gibberellin 20-oxidase (GA20ox) gene in tandem with the increased expression of Gibberellin 2-oxidase (GA2ox) genes in transgenic PvKN1 lines suggest that PvKN1 may exert regulatory effects via modulation of GA signaling. Furthermore, overexpression ofmore » PvKN1 altered the expression of cellulose and hemicellulose biosynthetic genes and increased sugar release efficiency in transgenic lines. Our findings demonstrated that switchgrass PvKN1 is a putative ortholog of maize KN1 that is linked to plant lignification and cell wall and development traits as a major regulatory gene. Therefore, targeted overexpression of PvKN1 in bioenergy feedstocks may provide one feasible strategy for reducing biomass recalcitrance and simultaneously improving plant growth characteristics.« less

Authors:
 [1];  [1];  [2];  [3];  [3];  [3];  [3];  [2]
  1. Univ. of Tennessee, Knoxville, TN (United States); Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
  2. Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); Samuel Roberts Noble Foundation, Ardmore, OK (United States)
  3. Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States); National Renewable Energy Lab. (NREL), Golden, CO (United States)
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)
OSTI Identifier:
1254112
Report Number(s):
NREL/JA-2700-66232
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 Issue: 7; Related Information: Frontiers in Plant Science; Journal ID: ISSN 1664-462X
Publisher:
Frontiers Research Foundation
Country of Publication:
United States
Language:
English
Subject:
09 BIOMASS FUELS; 59 BASIC BIOLOGICAL SCIENCES; gene regulation; lignin; gibberellin; cell walls; cellulose; hemicellulose; sugar release; biofuel

Citation Formats

Wuddineh, Wegi A., Mazarei, Mitra, Zhang, Ji -Yi, Turner, Geoffrey B., Sykes, Robert W., Decker, Stephen R., Davis, Mark F., Udvardi, Michael K., and C. Neal Stewart, Jr. Identification and overexpression of a knotted1-like transcription factor in switchgrass (Panicum virgatum L.) for lignocellulosic feedstock improvement. United States: N. p., 2016. Web. doi:10.3389/fpls.2016.00520.
Wuddineh, Wegi A., Mazarei, Mitra, Zhang, Ji -Yi, Turner, Geoffrey B., Sykes, Robert W., Decker, Stephen R., Davis, Mark F., Udvardi, Michael K., & C. Neal Stewart, Jr. Identification and overexpression of a knotted1-like transcription factor in switchgrass (Panicum virgatum L.) for lignocellulosic feedstock improvement. United States. doi:10.3389/fpls.2016.00520.
Wuddineh, Wegi A., Mazarei, Mitra, Zhang, Ji -Yi, Turner, Geoffrey B., Sykes, Robert W., Decker, Stephen R., Davis, Mark F., Udvardi, Michael K., and C. Neal Stewart, Jr. Thu . "Identification and overexpression of a knotted1-like transcription factor in switchgrass (Panicum virgatum L.) for lignocellulosic feedstock improvement". United States. doi:10.3389/fpls.2016.00520. https://www.osti.gov/servlets/purl/1254112.
@article{osti_1254112,
title = {Identification and overexpression of a knotted1-like transcription factor in switchgrass (Panicum virgatum L.) for lignocellulosic feedstock improvement},
author = {Wuddineh, Wegi A. and Mazarei, Mitra and Zhang, Ji -Yi and Turner, Geoffrey B. and Sykes, Robert W. and Decker, Stephen R. and Davis, Mark F. and Udvardi, Michael K. and C. Neal Stewart, Jr.},
abstractNote = {High biomass production and wide adaptation has made switchgrass (Panicum virgatum L.) an important candidate lignocellulosic bioenergy crop. One major limitation of this and other lignocellulosic feedstocks is the recalcitrance of complex carbohydrates to hydrolysis for conversion to biofuels. Lignin is the major contributor to recalcitrance as it limits the accessibility of cell wall carbohydrates to enzymatic breakdown into fermentable sugars. Therefore, genetic manipulation of the lignin biosynthesis pathway is one strategy to reduce recalcitrance. Here, we identified a switchgrass Knotted1 transcription factor, PvKN1, with the aim of genetically engineering switchgrass for reduced biomass recalcitrance for biofuel production. Gene expression of the endogenous PvKN1 gene was observed to be highest in young inflorescences and stems. Ectopic overexpression of PvKN1 in switchgrass altered growth, especially in early developmental stages. Transgenic lines had reduced expression of most lignin biosynthetic genes accompanied by a reduction in lignin content suggesting the involvement of PvKN1 in the broad regulation of the lignin biosynthesis pathway. Moreover, the reduced expression of the Gibberellin 20-oxidase (GA20ox) gene in tandem with the increased expression of Gibberellin 2-oxidase (GA2ox) genes in transgenic PvKN1 lines suggest that PvKN1 may exert regulatory effects via modulation of GA signaling. Furthermore, overexpression of PvKN1 altered the expression of cellulose and hemicellulose biosynthetic genes and increased sugar release efficiency in transgenic lines. Our findings demonstrated that switchgrass PvKN1 is a putative ortholog of maize KN1 that is linked to plant lignification and cell wall and development traits as a major regulatory gene. Therefore, targeted overexpression of PvKN1 in bioenergy feedstocks may provide one feasible strategy for reducing biomass recalcitrance and simultaneously improving plant growth characteristics.},
doi = {10.3389/fpls.2016.00520},
journal = {Frontiers in Plant Science},
issn = {1664-462X},
number = 7,
volume = 7,
place = {United States},
year = {2016},
month = {4}
}

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Works referenced in this record:

Coordinated Activation of Cellulose and Repression of Lignin Biosynthesis Pathways in Rice
journal, December 2010

  • Ambavaram, Madana M. R.; Krishnan, Arjun; Trijatmiko, Kurniawan R.
  • Plant Physiology, Vol. 155, Issue 2
  • DOI: 10.1104/pp.110.168641

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

The Populus homeobox gene ARBORKNOX2 regulates cell differentiation during secondary growth: KNOX gene regulation of secondary growth
journal, December 2009


The Making of a Compound Leaf: Genetic Manipulation of Leaf Architecture in Tomato
journal, March 1996


The Gibberellin Pathway Mediates KNOTTED1-Type Homeobox Function in Plants with Different Body Plans
journal, September 2002


A Comprehensive Classification and Evolutionary Analysis of Plant Homeobox Genes
journal, September 2009

  • Mukherjee, K.; Brocchieri, L.; Burglin, T. R.
  • Molecular Biology and Evolution, Vol. 26, Issue 12
  • DOI: 10.1093/molbev/msp201

Overexpression of Rice OSH Genes Induces Ectopic Shoots on Leaf Sheaths of Transgenic Rice Plants
journal, April 2000

  • Sentoku, Naoki; Sato, Yutaka; Matsuoka, Makoto
  • Developmental Biology, Vol. 220, Issue 2
  • DOI: 10.1006/dbio.2000.9624

A rice homeotic gene, OSH1 , causes unusual phenotypes in transgenic tobacco
journal, November 1993


Functional Analysis of the Conserved Domains of a Rice KNOX Homeodomain Protein, OSH15
journal, September 2001


The developmental gene Knotted-1 is a member of a maize homeobox gene family
journal, March 1991

  • Vollbrecht, Erik; Veit, Bruce; Sinha, Neelima
  • Nature, Vol. 350, Issue 6315
  • DOI: 10.1038/350241a0

Transcriptional Activation of Secondary Wall Biosynthesis by Rice and Maize NAC and MYB Transcription Factors
journal, September 2011

  • Zhong, R.; Lee, C.; McCarthy, R. L.
  • Plant and Cell Physiology, Vol. 52, Issue 10
  • DOI: 10.1093/pcp/pcr123

Analysis of the Competence to Respond to KNOTTED1 Activity in Arabidopsis Leaves Using a Steroid Induction System
journal, March 2003


ZmMYB31 directly represses maize lignin genes and redirects the phenylpropanoid metabolic flux: ZmMYB31 controls phenylpropanoids
journal, October 2010


GSDS: a gene structure display server
journal, January 2007


Sequence analysis and expression patterns divide the maize knotted1-like homeobox genes into two classes.
journal, December 1994


Silencing of 4-coumarate:coenzyme A ligase in switchgrass leads to reduced lignin content and improved fermentable sugar yields for biofuel production
journal, July 2011


The Path Forward for Biofuels and Biomaterials
journal, January 2006

  • Ragauskas, Arthur J.; Williams, Charlotte K.; Davison, Brian H.
  • Science, Vol. 311, Issue 5760, p. 484-489
  • DOI: 10.1126/science.1114736

Downregulation of Cinnamyl Alcohol Dehydrogenase (CAD) Leads to Improved Saccharification Efficiency in Switchgrass
journal, January 2011


Unraveling the KNOTTED1 regulatory network in maize meristems
journal, August 2012

  • Bolduc, N.; Yilmaz, A.; Mejia-Guerra, M. K.
  • Genes & Development, Vol. 26, Issue 15
  • DOI: 10.1101/gad.193433.112

KNOX genes: versatile regulators of plant development and diversity
journal, September 2010


Knotted1-like genes are strongly expressed in differentiated cell types in sunflower
journal, February 2003


Switchgrass (Panicum virgatum) possesses a divergent family of cinnamoyl CoA reductases with distinct biochemical properties
journal, September 2009


An Improved Tissue Culture System for Embryogenic Callus Production and Plant Regeneration in Switchgrass (Panicum virgatum L.)
journal, October 2009


How biotech can transform biofuels
journal, February 2008

  • Lynd, Lee R.; Laser, Mark S.; Bransby, David
  • Nature Biotechnology, Vol. 26, Issue 2, p. 169-172
  • DOI: 10.1038/nbt0208-169

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

Net energy of cellulosic ethanol from switchgrass
journal, January 2008

  • Schmer, M. R.; Vogel, K. P.; Mitchell, R. B.
  • Proceedings of the National Academy of Sciences, Vol. 105, Issue 2
  • DOI: 10.1073/pnas.0704767105

Phylogeny.fr: robust phylogenetic analysis for the non-specialist
journal, May 2008

  • Dereeper, A.; Guignon, V.; Blanc, G.
  • Nucleic Acids Research, Vol. 36, Issue Web Server
  • DOI: 10.1093/nar/gkn180

Unequal Redundancy in Maize knotted1 homeobox Genes
journal, November 2013


A Genomics Approach to Deciphering Lignin Biosynthesis in Switchgrass
journal, November 2013


A protocol for Agrobacterium-mediated transformation in rice
journal, December 2006

  • Nishimura, Asuka; Aichi, Ikuko; Matsuoka, Makoto
  • Nature Protocols, Vol. 1, Issue 6
  • DOI: 10.1038/nprot.2006.469

Transcriptional Regulation of Grass Secondary Cell Wall Biosynthesis: Playing Catch-Up with Arabidopsis thaliana
journal, January 2012


Over-expression of the AtGA2ox8 gene decreases the biomass accumulation and lignification in rapeseed (Brassica napus L.)
journal, July 2010

  • Zhao, Xiao-ying; Zhu, Deng-feng; Zhou, Bo
  • Journal of Zhejiang University SCIENCE B, Vol. 11, Issue 7
  • DOI: 10.1631/jzus.B1000161

Stress-Induced Cytokinin Synthesis Increases Drought Tolerance through the Coordinated Regulation of Carbon and Nitrogen Assimilation in Rice
journal, October 2013

  • Reguera, M.; Peleg, Z.; Abdel-Tawab, Y. M.
  • PLANT PHYSIOLOGY, Vol. 163, Issue 4
  • DOI: 10.1104/pp.113.227702

Plants to power: bioenergy to fuel the future
journal, August 2008


The NAC transcription factor OsSWN1 regulates secondary cell wall development in Oryza sativa
journal, January 2015


The Class II KNOX gene KNAT7 negatively regulates secondary wall formation in Arabidopsis and is functionally conserved in Populus
journal, January 2012


Complexity of the transcriptional network controlling secondary wall biosynthesis
journal, December 2014


THE ROLE OF KNOX GENES IN PLANT DEVELOPMENT
journal, November 2004


Functional characterization of the switchgrass (Panicum virgatum) R2R3-MYB transcription factor PvMYB4 for improvement of lignocellulosic feedstocks
journal, October 2011


Genetic manipulation of lignin reduces recalcitrance and improves ethanol production from switchgrass
journal, February 2011

  • Fu, Chunxiang; Mielenz, Jonathan R.; Xiao, Xirong
  • Proceedings of the National Academy of Sciences, Vol. 108, Issue 9, p. 3803-3808
  • DOI: 10.1073/pnas.1100310108

The Populus homeobox gene ARBORKNOX1 reveals overlapping mechanisms regulating the shoot apical meristem and the vascular cambium
journal, August 2006

  • Groover, Andrew T.; Mansfield, Shawn D.; DiFazio, Stephen P.
  • Plant Molecular Biology, Vol. 61, Issue 6
  • DOI: 10.1007/s11103-006-0059-y

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


High throughput Agrobacterium-mediated switchgrass transformation
journal, March 2011


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


A Revised Medium for Rapid Growth and Bio Assays with Tobacco Tissue Cultures
journal, July 1962


Genome-Wide Study of KNOX Regulatory Network Reveals Brassinosteroid Catabolic Genes Important for Shoot Meristem Function in Rice
journal, September 2014


KNAT1 induces lobed leaves with ectopic meristems when overexpressed in Arabidopsis.
journal, August 1996


Biolistic transformation of elite genotypes of switchgrass (Panicum virgatum L.)
journal, November 2013


Brassinosteroids can regulate cellulose biosynthesis by controlling the expression of CESA genes in Arabidopsis
journal, May 2011

  • Xie, Liqiong; Yang, Cangjing; Wang, Xuelu
  • Journal of Experimental Botany, Vol. 62, Issue 13
  • DOI: 10.1093/jxb/err164

A conceptual framework for maize leaf development
journal, September 1992


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


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

The Maize Transcription Factor KNOTTED1 Directly Regulates the Gibberellin Catabolism Gene ga2ox1
journal, June 2009


KNOX Action in Arabidopsis Is Mediated by Coordinate Regulation of Cytokinin and Gibberellin Activities
journal, September 2005


Expression of a rice homeobox gene causes altered morphology of transgenic plants.
journal, September 1993


Arabidopsis KNOXI Proteins Activate Cytokinin Biosynthesis
journal, September 2005


Gnarley1 Is a Dominant Mutation in the knox4 Homeobox Gene Affecting Cell Shape and Identity
journal, July 1999

  • Foster, Toshi; Yamaguchi, Judy; Wong, Bryan C.
  • The Plant Cell, Vol. 11, Issue 7
  • DOI: 10.2307/3870746

Peach [Prunus persica (L.) Batsch] KNOPE1, a class 1 KNOX orthologue to Arabidopsis BREVIPEDICELLUS/KNAT1, is misexpressed during hyperplasia of leaf curl disease
journal, February 2008

  • Testone, Giulio; Bruno, Leonardo; Condello, Emiliano
  • Journal of Experimental Botany, Vol. 59, Issue 2
  • DOI: 10.1093/jxb/erm317

MUSCLE: multiple sequence alignment with high accuracy and high throughput
journal, March 2004

  • Edgar, R. C.
  • Nucleic Acids Research, Vol. 32, Issue 5, p. 1792-1797
  • DOI: 10.1093/nar/gkh340

KNOX1 genes regulate lignin deposition and composition in monocots and dicots
journal, January 2013

  • Townsley, Brad T.; Sinha, Neelima R.; Kang, Julie
  • Frontiers in Plant Science, Vol. 4
  • DOI: 10.3389/fpls.2013.00121

Influence of antibiotic cefotaxime on somatic embryogenesis and plant regeneration in indica rice
journal, October 2006

  • Grewal, Deepinder; Gill, Raman; Gosal, Satbir Singh
  • Biotechnology Journal, Vol. 1, Issue 10
  • DOI: 10.1002/biot.200600139

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

Impact of Altered Gibberellin Metabolism on Biomass Accumulation, Lignin Biosynthesis, and Photosynthesis in Transgenic Tobacco Plants
journal, April 2004

  • Biemelt, Sophia; Tschiersch, Henning; Sonnewald, Uwe
  • Plant Physiology, Vol. 135, Issue 1
  • DOI: 10.1104/pp.103.036988

Rice Snl6, a Cinnamoyl-CoA Reductase-Like Gene Family Member, Is Required for NH1-Mediated Immunity to Xanthomonas oryzae pv. oryzae
journal, September 2010

  • Bart, Rebecca S.; Chern, Mawsheng; Vega-Sánchez, Miguel E.
  • PLoS Genetics, Vol. 6, Issue 9, Article No. e1001123
  • DOI: 10.1371/journal.pgen.1001123

Down-regulation of the caffeic acid O-methyltransferase gene in switchgrass reveals a novel monolignol analog
journal, January 2012

  • Tschaplinski, Timothy J.; Standaert, Robert F.; Engle, Nancy L.
  • Biotechnology for Biofuels, Vol. 5, Issue 1
  • DOI: 10.1186/1754-6834-5-71

MYB58 and MYB63 Are Transcriptional Activators of the Lignin Biosynthetic Pathway during Secondary Cell Wall Formation in Arabidopsis
journal, January 2009


    Works referencing / citing this record:

    Suberin and hemicellulose in sugarcane cell wall architecture and crop digestibility: A biotechnological perspective
    journal, February 2019

    • Figueiredo, Raquel; Araújo, Pedro; Llerena, Juan Pablo P.
    • Food and Energy Security, Vol. 8, Issue 3
    • DOI: 10.1002/fes3.163

    Genetic engineering of grass cell wall polysaccharides for biorefining
    journal, June 2017

    • Bhatia, Rakesh; Gallagher, Joe A.; Gomez, Leonardo D.
    • Plant Biotechnology Journal, Vol. 15, Issue 9
    • DOI: 10.1111/pbi.12764

    Overexpression of a rice BAHD acyltransferase gene in switchgrass (Panicum virgatum L.) enhances saccharification
    journal, September 2018


    Current Models for Transcriptional Regulation of Secondary Cell Wall Biosynthesis in Grasses
    journal, April 2018