Downregulation of GAUT12 in Populus deltoides by RNA silencing results in reduced recalcitrance, increased growth and reduced xylan and pectin in a woody biofuel feedstock. Biotechnology For Biofuels
Abstract
Background The inherent recalcitrance of woody bioenergy feedstocks is a major challenge for their use as a source of second-generation biofuel. Secondary cell walls that constitute the majority of hardwood biomass are rich in cellulose, xylan, and lignin. The interactions among these polymers prevent facile accessibility and deconstruction by enzymes and chemicals. Plant biomass that can with minimal pretreatment be degraded into sugars is required to produce renewable biofuels in a cost-effective manner. Results GAUT12/IRX8 is a putative glycosyltransferase proposed to be involved in secondary cell wall glucuronoxylan and/or pectin biosynthesis based on concomitant reductions of both xylan and the pectin homogalacturonan (HG) in Arabidopsis irx8 mutants. Two GAUT12 homologs exist in Populus trichocarpa, PtGAUT12.1 and PtGAUT12.2. Knockdown expression of both genes simultaneously has been shown to reduce xylan content in Populus wood. We tested the proposition that RNA interference (RNAi) downregulation of GAUT12.1 alone would lead to increased sugar release in Populus wood, that is, reduced recalcitrance, based on the hypothesis that GAUT12 synthesizes a wall structure required for deposition of xylan and that cell walls with less xylan and/or modified cell wall architecture would have reduced recalcitrance. Using an RNAi approach, we generated 11 Populus deltoides transgenic linesmore »
- Authors:
-
- Univ. of Georgia, Athens, GA (United States); Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). BioEnergy Science Center (BESC)
- Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). BioEnergy Science Center (BESC)
- Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). BioEnergy Science Center (BESC); ArborGen Inc., Ridgeville, SC (United States)
- Univ. of Georgia, Athens, GA (United States)
- Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). BioEnergy Science Center (BESC); National Renewable Energy Lab. (NREL), Golden, CO (United States)
- Publication Date:
- Research Org.:
- Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States); National Renewable Energy Laboratory (NREL), Golden, CO (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Biological and Environmental Research (BER)
- OSTI Identifier:
- 1265706
- Alternate Identifier(s):
- OSTI ID: 1220619
- Report Number(s):
- NREL/JA-2700-62639
Journal ID: ISSN 1754-6834; KP1601050; ERKP695
- Grant/Contract Number:
- AC05-00OR22725; PS02-06ER64304; FG02-93ER20097
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Biotechnology for Biofuels
- Additional Journal Information:
- Journal Volume: 8; Journal Issue: 1; Journal ID: ISSN 1754-6834
- Publisher:
- BioMed Central
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 09 BIOMASS FUELS; 59 BASIC BIOLOGICAL SCIENCES; biofuels; growth; pectin; Populus; saccharification; secondary cell wall; xylan; wood development
Citation Formats
Biswal, Ajaya K., Hao, Zhangying, Pattathil, Sivakumar, Yang, Xiaohan, Winkeler, Kim, Collins, Cassandra, Mohanty, Sushree S., Richardson, Elizabeth A., Gelineo-Albersheim, Ivana, Hunt, Kimberly, Ryno, David, Sykes, Robert W., Turner, Geoffrey B., Ziebell, Angela, Gjersing, Erica, Lukowitz, Wolfgang, Davis, Mark F., Decker, Stephen R., Hahn, Michael G., and Mohnen, Debra A. Downregulation of GAUT12 in Populus deltoides by RNA silencing results in reduced recalcitrance, increased growth and reduced xylan and pectin in a woody biofuel feedstock. Biotechnology For Biofuels. United States: N. p., 2015.
Web. doi:10.1186/s13068-015-0218-y.
Biswal, Ajaya K., Hao, Zhangying, Pattathil, Sivakumar, Yang, Xiaohan, Winkeler, Kim, Collins, Cassandra, Mohanty, Sushree S., Richardson, Elizabeth A., Gelineo-Albersheim, Ivana, Hunt, Kimberly, Ryno, David, Sykes, Robert W., Turner, Geoffrey B., Ziebell, Angela, Gjersing, Erica, Lukowitz, Wolfgang, Davis, Mark F., Decker, Stephen R., Hahn, Michael G., & Mohnen, Debra A. Downregulation of GAUT12 in Populus deltoides by RNA silencing results in reduced recalcitrance, increased growth and reduced xylan and pectin in a woody biofuel feedstock. Biotechnology For Biofuels. United States. https://doi.org/10.1186/s13068-015-0218-y
Biswal, Ajaya K., Hao, Zhangying, Pattathil, Sivakumar, Yang, Xiaohan, Winkeler, Kim, Collins, Cassandra, Mohanty, Sushree S., Richardson, Elizabeth A., Gelineo-Albersheim, Ivana, Hunt, Kimberly, Ryno, David, Sykes, Robert W., Turner, Geoffrey B., Ziebell, Angela, Gjersing, Erica, Lukowitz, Wolfgang, Davis, Mark F., Decker, Stephen R., Hahn, Michael G., and Mohnen, Debra A. Thu .
"Downregulation of GAUT12 in Populus deltoides by RNA silencing results in reduced recalcitrance, increased growth and reduced xylan and pectin in a woody biofuel feedstock. Biotechnology For Biofuels". United States. https://doi.org/10.1186/s13068-015-0218-y. https://www.osti.gov/servlets/purl/1265706.
@article{osti_1265706,
title = {Downregulation of GAUT12 in Populus deltoides by RNA silencing results in reduced recalcitrance, increased growth and reduced xylan and pectin in a woody biofuel feedstock. Biotechnology For Biofuels},
author = {Biswal, Ajaya K. and Hao, Zhangying and Pattathil, Sivakumar and Yang, Xiaohan and Winkeler, Kim and Collins, Cassandra and Mohanty, Sushree S. and Richardson, Elizabeth A. and Gelineo-Albersheim, Ivana and Hunt, Kimberly and Ryno, David and Sykes, Robert W. and Turner, Geoffrey B. and Ziebell, Angela and Gjersing, Erica and Lukowitz, Wolfgang and Davis, Mark F. and Decker, Stephen R. and Hahn, Michael G. and Mohnen, Debra A.},
abstractNote = {Background The inherent recalcitrance of woody bioenergy feedstocks is a major challenge for their use as a source of second-generation biofuel. Secondary cell walls that constitute the majority of hardwood biomass are rich in cellulose, xylan, and lignin. The interactions among these polymers prevent facile accessibility and deconstruction by enzymes and chemicals. Plant biomass that can with minimal pretreatment be degraded into sugars is required to produce renewable biofuels in a cost-effective manner. Results GAUT12/IRX8 is a putative glycosyltransferase proposed to be involved in secondary cell wall glucuronoxylan and/or pectin biosynthesis based on concomitant reductions of both xylan and the pectin homogalacturonan (HG) in Arabidopsis irx8 mutants. Two GAUT12 homologs exist in Populus trichocarpa, PtGAUT12.1 and PtGAUT12.2. Knockdown expression of both genes simultaneously has been shown to reduce xylan content in Populus wood. We tested the proposition that RNA interference (RNAi) downregulation of GAUT12.1 alone would lead to increased sugar release in Populus wood, that is, reduced recalcitrance, based on the hypothesis that GAUT12 synthesizes a wall structure required for deposition of xylan and that cell walls with less xylan and/or modified cell wall architecture would have reduced recalcitrance. Using an RNAi approach, we generated 11 Populus deltoides transgenic lines with 50 to 67% reduced PdGAUT12.1 transcript expression compared to wild type (WT) and vector controls. Ten of the eleven RNAi lines yielded 4 to 8% greater glucose release upon enzymatic saccharification than the controls. The PdGAUT12.1 knockdown (PdGAUT12.1-KD) lines also displayed 12 to 52% and 12 to 44% increased plant height and radial stem diameter, respectively, compared to the controls. Knockdown of PdGAUT12.1 resulted in a 25 to 47% reduction in galacturonic acid and 17 to 30% reduction in xylose without affecting total lignin content, revealing that in Populus wood as in Arabidopsis, GAUT12 affects both pectin and xylan formation. Analyses of the sugars present in sequential cell wall extracts revealed a reduction of glucuronoxylan and pectic HG and rhamnogalacturonan in extracts from PdGAUT12.1-KD lines. Conclusions The results show that downregulation of GAUT12.1 leads to a reduction in a population of xylan and pectin during wood formation and to reduced recalcitrance, more easily extractable cell walls, and increased growth in Populus.},
doi = {10.1186/s13068-015-0218-y},
journal = {Biotechnology for Biofuels},
number = 1,
volume = 8,
place = {United States},
year = {Thu Mar 12 00:00:00 EDT 2015},
month = {Thu Mar 12 00:00:00 EDT 2015}
}
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Figures / Tables:
Figure 1: Phylogenetic tree of GAUTs in Arabidopsis thaliana and Populus trichocarpa. Phylogenetic tree showing the relationship between amino acid sequences of the GAUT Protein Family of Arabidopsis thaliana (TAIR10) and Populus trichocarpa (Phytozome 8.0). The tree was constructed by the Neighbor-Joining method using MEGA5. POPTR_0001s44250 is named GAUT12.1 inmore »
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