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Systematic approaches to C-lignin engineering in Medicago truncatula

BACKGROUND: C-lignin is a homopolymer of caffeyl alcohol present in the seed coats of a variety of plant species including vanilla orchid, various cacti, and the ornamental plant Cleome hassleriana. Because of its unique chemical and physical properties, there is considerable interest in engineering...

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Autores principales: Ha, Chan Man, Escamilla-Trevino, Luis, Zhuo, Chunliu, Pu, Yunqiao, Bryant, Nathan, Ragauskas, Arthur J., Xiao, Xirong, Li, Ying, Chen, Fang, Dixon, Richard A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10262568/
https://www.ncbi.nlm.nih.gov/pubmed/37308891
http://dx.doi.org/10.1186/s13068-023-02339-7
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author Ha, Chan Man
Escamilla-Trevino, Luis
Zhuo, Chunliu
Pu, Yunqiao
Bryant, Nathan
Ragauskas, Arthur J.
Xiao, Xirong
Li, Ying
Chen, Fang
Dixon, Richard A.
author_facet Ha, Chan Man
Escamilla-Trevino, Luis
Zhuo, Chunliu
Pu, Yunqiao
Bryant, Nathan
Ragauskas, Arthur J.
Xiao, Xirong
Li, Ying
Chen, Fang
Dixon, Richard A.
author_sort Ha, Chan Man
collection PubMed
description BACKGROUND: C-lignin is a homopolymer of caffeyl alcohol present in the seed coats of a variety of plant species including vanilla orchid, various cacti, and the ornamental plant Cleome hassleriana. Because of its unique chemical and physical properties, there is considerable interest in engineering C-lignin into the cell walls of bioenergy crops as a high-value co-product of bioprocessing. We have used information from a transcriptomic analysis of developing C. hassleriana seed coats to suggest strategies for engineering C-lignin in a heterologous system, using hairy roots of the model legume Medicago truncatula. RESULTS: We systematically tested strategies for C-lignin engineering using a combination of gene overexpression and RNAi-mediated knockdown in the caffeic acid/5-hydroxy coniferaldehyde 3/5-O-methyltransferase (comt) mutant background, monitoring the outcomes by analysis of lignin composition and profiling of monolignol pathway metabolites. In all cases, C-lignin accumulation required strong down-regulation of caffeoyl CoA 3-O-methyltransferase (CCoAOMT) paired with loss of function of COMT. Overexpression of the Selaginella moellendorffii ferulate 5-hydroxylase (SmF5H) gene in comt mutant hairy roots resulted in lines that unexpectedly accumulated high levels of S-lignin. CONCLUSION: C-Lignin accumulation of up to 15% of total lignin in lines with the greatest reduction in CCoAOMT expression required the strong down-regulation of both COMT and CCoAOMT, but did not require expression of a heterologous laccase, cinnamyl alcohol dehydrogenase (CAD) or cinnamoyl CoA reductase (CCR) with preference for 3,4-dihydroxy-substituted substrates in M. truncatula hairy roots. Cell wall fractionation studies suggested that the engineered C-units are not present in a heteropolymer with the bulk of the G-lignin. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13068-023-02339-7.
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spelling pubmed-102625682023-06-15 Systematic approaches to C-lignin engineering in Medicago truncatula Ha, Chan Man Escamilla-Trevino, Luis Zhuo, Chunliu Pu, Yunqiao Bryant, Nathan Ragauskas, Arthur J. Xiao, Xirong Li, Ying Chen, Fang Dixon, Richard A. Biotechnol Biofuels Bioprod Research BACKGROUND: C-lignin is a homopolymer of caffeyl alcohol present in the seed coats of a variety of plant species including vanilla orchid, various cacti, and the ornamental plant Cleome hassleriana. Because of its unique chemical and physical properties, there is considerable interest in engineering C-lignin into the cell walls of bioenergy crops as a high-value co-product of bioprocessing. We have used information from a transcriptomic analysis of developing C. hassleriana seed coats to suggest strategies for engineering C-lignin in a heterologous system, using hairy roots of the model legume Medicago truncatula. RESULTS: We systematically tested strategies for C-lignin engineering using a combination of gene overexpression and RNAi-mediated knockdown in the caffeic acid/5-hydroxy coniferaldehyde 3/5-O-methyltransferase (comt) mutant background, monitoring the outcomes by analysis of lignin composition and profiling of monolignol pathway metabolites. In all cases, C-lignin accumulation required strong down-regulation of caffeoyl CoA 3-O-methyltransferase (CCoAOMT) paired with loss of function of COMT. Overexpression of the Selaginella moellendorffii ferulate 5-hydroxylase (SmF5H) gene in comt mutant hairy roots resulted in lines that unexpectedly accumulated high levels of S-lignin. CONCLUSION: C-Lignin accumulation of up to 15% of total lignin in lines with the greatest reduction in CCoAOMT expression required the strong down-regulation of both COMT and CCoAOMT, but did not require expression of a heterologous laccase, cinnamyl alcohol dehydrogenase (CAD) or cinnamoyl CoA reductase (CCR) with preference for 3,4-dihydroxy-substituted substrates in M. truncatula hairy roots. Cell wall fractionation studies suggested that the engineered C-units are not present in a heteropolymer with the bulk of the G-lignin. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13068-023-02339-7. BioMed Central 2023-06-12 /pmc/articles/PMC10262568/ /pubmed/37308891 http://dx.doi.org/10.1186/s13068-023-02339-7 Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research
Ha, Chan Man
Escamilla-Trevino, Luis
Zhuo, Chunliu
Pu, Yunqiao
Bryant, Nathan
Ragauskas, Arthur J.
Xiao, Xirong
Li, Ying
Chen, Fang
Dixon, Richard A.
Systematic approaches to C-lignin engineering in Medicago truncatula
title Systematic approaches to C-lignin engineering in Medicago truncatula
title_full Systematic approaches to C-lignin engineering in Medicago truncatula
title_fullStr Systematic approaches to C-lignin engineering in Medicago truncatula
title_full_unstemmed Systematic approaches to C-lignin engineering in Medicago truncatula
title_short Systematic approaches to C-lignin engineering in Medicago truncatula
title_sort systematic approaches to c-lignin engineering in medicago truncatula
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10262568/
https://www.ncbi.nlm.nih.gov/pubmed/37308891
http://dx.doi.org/10.1186/s13068-023-02339-7
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