A gene-editing/complementation strategy for tissue-specific lignin reduction while preserving biomass yield
Abstract Background Lignification of secondary cell walls is a major factor conferring recalcitrance of lignocellulosic biomass to deconstruction for fuels and chemicals. Genetic modification can reduce lignin content and enhance saccharification efficiency, but usually at the cost of moderate-to-se...
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| Format: | Article |
| Language: | English |
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BMC
2021-09-01
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| Series: | Biotechnology for Biofuels |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13068-021-02026-5 |
| _version_ | 1818263893708898304 |
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| author | Hasi Yu Chang Liu Richard A. Dixon |
| author_facet | Hasi Yu Chang Liu Richard A. Dixon |
| author_sort | Hasi Yu |
| collection | DOAJ |
| description | Abstract Background Lignification of secondary cell walls is a major factor conferring recalcitrance of lignocellulosic biomass to deconstruction for fuels and chemicals. Genetic modification can reduce lignin content and enhance saccharification efficiency, but usually at the cost of moderate-to-severe growth penalties. We have developed a method, using a single DNA construct that uses CRISPR–Cas9 gene editing to knock-out expression of an endogenous gene of lignin monomer biosynthesis while at the same time expressing a modified version of the gene’s open reading frame that escapes cutting by the Cas9 system and complements the introduced mutation in a tissue-specific manner. Results Expressing the complementing open reading frame in vessels allows for the regeneration of Arabidopsis plants with reduced lignin, wild-type biomass yield, and up to fourfold enhancement of cell wall sugar yield per plant. The above phenotypes are seen in both homozygous and bi-allelic heterozygous T1 lines, and are stable over at least four generations. Conclusions The method provides a rapid approach for generating reduced lignin trees or crops with one single transformation event, and, paired with a range of tissue-specific promoters, provides a general strategy for optimizing loss-of-function traits that are associated with growth penalties. This method should be applicable to any plant species in which transformation and gene editing are feasible and validated vessel-specific promoters are available. |
| first_indexed | 2024-12-12T19:26:15Z |
| format | Article |
| id | doaj.art-771349e0dbc346a2b6ef8dbafc560b80 |
| institution | Directory Open Access Journal |
| issn | 1754-6834 |
| language | English |
| last_indexed | 2024-12-12T19:26:15Z |
| publishDate | 2021-09-01 |
| publisher | BMC |
| record_format | Article |
| series | Biotechnology for Biofuels |
| spelling | doaj.art-771349e0dbc346a2b6ef8dbafc560b802022-12-22T00:14:30ZengBMCBiotechnology for Biofuels1754-68342021-09-0114111210.1186/s13068-021-02026-5A gene-editing/complementation strategy for tissue-specific lignin reduction while preserving biomass yieldHasi Yu0Chang Liu1Richard A. Dixon2BioDiscovery Institute and Department of Biological Sciences, University of North TexasBioDiscovery Institute and Department of Biological Sciences, University of North TexasBioDiscovery Institute and Department of Biological Sciences, University of North TexasAbstract Background Lignification of secondary cell walls is a major factor conferring recalcitrance of lignocellulosic biomass to deconstruction for fuels and chemicals. Genetic modification can reduce lignin content and enhance saccharification efficiency, but usually at the cost of moderate-to-severe growth penalties. We have developed a method, using a single DNA construct that uses CRISPR–Cas9 gene editing to knock-out expression of an endogenous gene of lignin monomer biosynthesis while at the same time expressing a modified version of the gene’s open reading frame that escapes cutting by the Cas9 system and complements the introduced mutation in a tissue-specific manner. Results Expressing the complementing open reading frame in vessels allows for the regeneration of Arabidopsis plants with reduced lignin, wild-type biomass yield, and up to fourfold enhancement of cell wall sugar yield per plant. The above phenotypes are seen in both homozygous and bi-allelic heterozygous T1 lines, and are stable over at least four generations. Conclusions The method provides a rapid approach for generating reduced lignin trees or crops with one single transformation event, and, paired with a range of tissue-specific promoters, provides a general strategy for optimizing loss-of-function traits that are associated with growth penalties. This method should be applicable to any plant species in which transformation and gene editing are feasible and validated vessel-specific promoters are available.https://doi.org/10.1186/s13068-021-02026-5SaccharificationLignin modificationGene editingCRISPRCinnamoyl CoA reductase (CCR1) |
| spellingShingle | Hasi Yu Chang Liu Richard A. Dixon A gene-editing/complementation strategy for tissue-specific lignin reduction while preserving biomass yield Biotechnology for Biofuels Saccharification Lignin modification Gene editing CRISPR Cinnamoyl CoA reductase (CCR1) |
| title | A gene-editing/complementation strategy for tissue-specific lignin reduction while preserving biomass yield |
| title_full | A gene-editing/complementation strategy for tissue-specific lignin reduction while preserving biomass yield |
| title_fullStr | A gene-editing/complementation strategy for tissue-specific lignin reduction while preserving biomass yield |
| title_full_unstemmed | A gene-editing/complementation strategy for tissue-specific lignin reduction while preserving biomass yield |
| title_short | A gene-editing/complementation strategy for tissue-specific lignin reduction while preserving biomass yield |
| title_sort | gene editing complementation strategy for tissue specific lignin reduction while preserving biomass yield |
| topic | Saccharification Lignin modification Gene editing CRISPR Cinnamoyl CoA reductase (CCR1) |
| url | https://doi.org/10.1186/s13068-021-02026-5 |
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