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 »
- Authors:
-
[2];
[2];
[7];
- Univ. of Tennessee, Knoxville, TN (United States). Department of Plant Sciences; Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). BioEnergy Science Center
- Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). BioEnergy Science Center and Biosciences Division, Joint Institute for Biological Science
- Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). BioEnergy Science Center; Univ. of Georgia, Athens, GA (United States). Complex Carbohydrate Research Center
- Univ. of Tennessee, Knoxville, TN (United States). Center for Renewable Carbon
- 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
- Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). BioEnergy Science Center; Noble Research Institute, Ardmore, OK (United States)
- National Renewable Energy Lab. (NREL), Golden, CO (United States)
- 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 Laboratory (NREL), Golden, CO (United States); Oak Ridge National Laboratory (ORNL), Oak Ridge, TN (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Biological and Environmental Research (BER)
- 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. https://doi.org/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. https://doi.org/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 = {Fri Aug 03 00:00:00 EDT 2018},
month = {Fri Aug 03 00:00:00 EDT 2018}
}
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