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Title: Engineering temporal accumulation of a low recalcitrance polysaccharide leads to increased C6 sugar content in plant cell walls

Abstract

Reduced cell wall recalcitrance and increased C6 monosaccharide content are desirable traits for future biofuel crops, as long as these biomass modifications do not significantly alter normal growth and development. Mixed-linkage glucan (MLG), a cell wall polysaccharide only present in grasses and related species among flowering plants, is comprised of glucose monomers linked by both β-1,3 and β-1,4 bonds. Previous data have shown that constitutive production of MLG in barley (Hordeum vulgare) severely compromises growth and development. Here, we used spatio-temporal strategies to engineer Arabidopsis thaliana plants to accumulate significant amounts of MLG in the cell wall by expressing the rice CslF6 MLG synthase using secondary cell wall and senescence-associated promoters. Results using secondary wall promoters were suboptimal. When the rice MLG synthase was expressed under the control of a senescence-associated promoter, we obtained up to four times more glucose in the matrix cell wall fraction and up to a 42% increase in saccharification compared to control lines. Importantly, these plants grew and developed normally. The induction of MLG deposition at senescence correlated with an increase of gluconic acid in cell wall extracts of transgenic plants in contrast to the other approaches presented in this study. MLG produced in Arabidopsismore » has an altered structure compared to the grass glucan, which likely affects its solubility, while its molecular size is unaffected. The induction of cell wall polysaccharide biosynthesis in senescing tissues offers a novel engineering alternative to enhance cell wall properties of lignocellulosic biofuel crops.« less

Authors:
ORCiD logo [1];  [1];  [1];  [1];  [1];  [1];  [1];  [2];  [3];  [1];  [4];  [5];  [1];  [6]
  1. Joint BioEnergy Institute and Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley CA USA
  2. Department of Plant and Environmental Sciences, University of Copenhagen, Frederiksberg C Denmark
  3. Joint BioEnergy Institute and Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley CA USA; Department of Plant and Environmental Sciences, University of Copenhagen, Frederiksberg C Denmark
  4. Joint BioEnergy Institute and Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley CA USA; Department of Chemical and Biomolecular Engineering, and Department of Bioengineering, University of California, Berkeley CA USA
  5. Joint BioEnergy Institute and Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley CA USA; Department of Plant and Microbial Biology, University of California, Berkeley CA USA
  6. Joint BioEnergy Institute and Physical Biosciences Division, Lawrence Berkeley National Laboratory, Berkeley CA USA; Department of Plant Pathology and the Genome Center, University of California, Davis CA USA
Publication Date:
Research Org.:
Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
OSTI Identifier:
1407289
DOE Contract Number:  
AC02-05CH11231
Resource Type:
Journal Article
Journal Name:
Plant Biotechnology Journal
Additional Journal Information:
Journal Volume: 13; Journal Issue: 7; Journal ID: ISSN 1467-7644
Publisher:
Society for Experimental Biology; Association of Applied Biology
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Vega-Sánchez, Miguel E., Loqué, Dominique, Lao, Jeemeng, Catena, Michela, Verhertbruggen, Yves, Herter, Thomas, Yang, Fan, Harholt, Jesper, Ebert, Berit, Baidoo, Edward E. K., Keasling, Jay D., Scheller, Henrik V., Heazlewood, Joshua L., and Ronald, Pamela C. Engineering temporal accumulation of a low recalcitrance polysaccharide leads to increased C6 sugar content in plant cell walls. United States: N. p., 2015. Web. doi:10.1111/pbi.12326.
Vega-Sánchez, Miguel E., Loqué, Dominique, Lao, Jeemeng, Catena, Michela, Verhertbruggen, Yves, Herter, Thomas, Yang, Fan, Harholt, Jesper, Ebert, Berit, Baidoo, Edward E. K., Keasling, Jay D., Scheller, Henrik V., Heazlewood, Joshua L., & Ronald, Pamela C. Engineering temporal accumulation of a low recalcitrance polysaccharide leads to increased C6 sugar content in plant cell walls. United States. doi:10.1111/pbi.12326.
Vega-Sánchez, Miguel E., Loqué, Dominique, Lao, Jeemeng, Catena, Michela, Verhertbruggen, Yves, Herter, Thomas, Yang, Fan, Harholt, Jesper, Ebert, Berit, Baidoo, Edward E. K., Keasling, Jay D., Scheller, Henrik V., Heazlewood, Joshua L., and Ronald, Pamela C. Wed . "Engineering temporal accumulation of a low recalcitrance polysaccharide leads to increased C6 sugar content in plant cell walls". United States. doi:10.1111/pbi.12326.
@article{osti_1407289,
title = {Engineering temporal accumulation of a low recalcitrance polysaccharide leads to increased C6 sugar content in plant cell walls},
author = {Vega-Sánchez, Miguel E. and Loqué, Dominique and Lao, Jeemeng and Catena, Michela and Verhertbruggen, Yves and Herter, Thomas and Yang, Fan and Harholt, Jesper and Ebert, Berit and Baidoo, Edward E. K. and Keasling, Jay D. and Scheller, Henrik V. and Heazlewood, Joshua L. and Ronald, Pamela C.},
abstractNote = {Reduced cell wall recalcitrance and increased C6 monosaccharide content are desirable traits for future biofuel crops, as long as these biomass modifications do not significantly alter normal growth and development. Mixed-linkage glucan (MLG), a cell wall polysaccharide only present in grasses and related species among flowering plants, is comprised of glucose monomers linked by both β-1,3 and β-1,4 bonds. Previous data have shown that constitutive production of MLG in barley (Hordeum vulgare) severely compromises growth and development. Here, we used spatio-temporal strategies to engineer Arabidopsis thaliana plants to accumulate significant amounts of MLG in the cell wall by expressing the rice CslF6 MLG synthase using secondary cell wall and senescence-associated promoters. Results using secondary wall promoters were suboptimal. When the rice MLG synthase was expressed under the control of a senescence-associated promoter, we obtained up to four times more glucose in the matrix cell wall fraction and up to a 42% increase in saccharification compared to control lines. Importantly, these plants grew and developed normally. The induction of MLG deposition at senescence correlated with an increase of gluconic acid in cell wall extracts of transgenic plants in contrast to the other approaches presented in this study. MLG produced in Arabidopsis has an altered structure compared to the grass glucan, which likely affects its solubility, while its molecular size is unaffected. The induction of cell wall polysaccharide biosynthesis in senescing tissues offers a novel engineering alternative to enhance cell wall properties of lignocellulosic biofuel crops.},
doi = {10.1111/pbi.12326},
journal = {Plant Biotechnology Journal},
issn = {1467-7644},
number = 7,
volume = 13,
place = {United States},
year = {2015},
month = {1}
}

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