VvEPFL9-1 Knock-Out via CRISPR/Cas9 Reduces Stomatal Density in Grapevine
Epidermal Patterning Factor Like 9 (EPFL9), also known as STOMAGEN, is a cysteine-rich peptide that induces stomata formation in vascular plants, acting antagonistically to other epidermal patterning factors (EPF1, EPF2). In grapevine there are two EPFL9 genes, EPFL9-1 and EPFL9-2 sharing 82% identi...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-05-01
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| Series: | Frontiers in Plant Science |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fpls.2022.878001/full |
| _version_ | 1811235378343968768 |
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| author | Molly Clemens Molly Clemens Molly Clemens Michele Faralli Jorge Lagreze Luana Bontempo Stefano Piazza Claudio Varotto Mickael Malnoy Walter Oechel Walter Oechel Annapaola Rizzoli Lorenza Dalla Costa |
| author_facet | Molly Clemens Molly Clemens Molly Clemens Michele Faralli Jorge Lagreze Luana Bontempo Stefano Piazza Claudio Varotto Mickael Malnoy Walter Oechel Walter Oechel Annapaola Rizzoli Lorenza Dalla Costa |
| author_sort | Molly Clemens |
| collection | DOAJ |
| description | Epidermal Patterning Factor Like 9 (EPFL9), also known as STOMAGEN, is a cysteine-rich peptide that induces stomata formation in vascular plants, acting antagonistically to other epidermal patterning factors (EPF1, EPF2). In grapevine there are two EPFL9 genes, EPFL9-1 and EPFL9-2 sharing 82% identity at protein level in the mature functional C-terminal domain. In this study, CRISPR/Cas9 system was applied to functionally characterize VvEPFL9-1 in ‘Sugraone’, a highly transformable genotype. A set of plants, regenerated after gene transfer in embryogenic calli via Agrobacterium tumefaciens, were selected for evaluation. For many lines, the editing profile in the target site displayed a range of mutations mainly causing frameshift in the coding sequence or affecting the second cysteine residue. The analysis of stomata density revealed that in edited plants the number of stomata was significantly reduced compared to control, demonstrating for the first time the role of EPFL9 in a perennial fruit crop. Three edited lines were then assessed for growth, photosynthesis, stomatal conductance, and water use efficiency in experiments carried out at different environmental conditions. Intrinsic water-use efficiency was improved in edited lines compared to control, indicating possible advantages in reducing stomatal density under future environmental drier scenarios. Our results show the potential of manipulating stomatal density for optimizing grapevine adaptation under changing climate conditions. |
| first_indexed | 2024-04-12T11:50:54Z |
| format | Article |
| id | doaj.art-d53b311dca554723935bfa14db930a29 |
| institution | Directory Open Access Journal |
| issn | 1664-462X |
| language | English |
| last_indexed | 2024-04-12T11:50:54Z |
| publishDate | 2022-05-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Plant Science |
| spelling | doaj.art-d53b311dca554723935bfa14db930a292022-12-22T03:34:11ZengFrontiers Media S.A.Frontiers in Plant Science1664-462X2022-05-011310.3389/fpls.2022.878001878001VvEPFL9-1 Knock-Out via CRISPR/Cas9 Reduces Stomatal Density in GrapevineMolly Clemens0Molly Clemens1Molly Clemens2Michele Faralli3Jorge Lagreze4Luana Bontempo5Stefano Piazza6Claudio Varotto7Mickael Malnoy8Walter Oechel9Walter Oechel10Annapaola Rizzoli11Lorenza Dalla Costa12Research and Innovation Centre, Fondazione Edmund Mach, San Michele all’Adige, ItalyGlobal Change Research Group, San Diego State University, San Diego, CA, United StatesDepartment of Viticulture and Enology, University of California Davis, Davis, CA, United StatesResearch and Innovation Centre, Fondazione Edmund Mach, San Michele all’Adige, ItalyResearch and Innovation Centre, Fondazione Edmund Mach, San Michele all’Adige, ItalyResearch and Innovation Centre, Fondazione Edmund Mach, San Michele all’Adige, ItalyResearch and Innovation Centre, Fondazione Edmund Mach, San Michele all’Adige, ItalyResearch and Innovation Centre, Fondazione Edmund Mach, San Michele all’Adige, ItalyResearch and Innovation Centre, Fondazione Edmund Mach, San Michele all’Adige, ItalyGlobal Change Research Group, San Diego State University, San Diego, CA, United StatesDepartment of Geography, University of Exeter, Exeter, United KingdomResearch and Innovation Centre, Fondazione Edmund Mach, San Michele all’Adige, ItalyResearch and Innovation Centre, Fondazione Edmund Mach, San Michele all’Adige, ItalyEpidermal Patterning Factor Like 9 (EPFL9), also known as STOMAGEN, is a cysteine-rich peptide that induces stomata formation in vascular plants, acting antagonistically to other epidermal patterning factors (EPF1, EPF2). In grapevine there are two EPFL9 genes, EPFL9-1 and EPFL9-2 sharing 82% identity at protein level in the mature functional C-terminal domain. In this study, CRISPR/Cas9 system was applied to functionally characterize VvEPFL9-1 in ‘Sugraone’, a highly transformable genotype. A set of plants, regenerated after gene transfer in embryogenic calli via Agrobacterium tumefaciens, were selected for evaluation. For many lines, the editing profile in the target site displayed a range of mutations mainly causing frameshift in the coding sequence or affecting the second cysteine residue. The analysis of stomata density revealed that in edited plants the number of stomata was significantly reduced compared to control, demonstrating for the first time the role of EPFL9 in a perennial fruit crop. Three edited lines were then assessed for growth, photosynthesis, stomatal conductance, and water use efficiency in experiments carried out at different environmental conditions. Intrinsic water-use efficiency was improved in edited lines compared to control, indicating possible advantages in reducing stomatal density under future environmental drier scenarios. Our results show the potential of manipulating stomatal density for optimizing grapevine adaptation under changing climate conditions.https://www.frontiersin.org/articles/10.3389/fpls.2022.878001/fullVitis viniferastomatagenome editingclimate changewater-use efficiency |
| spellingShingle | Molly Clemens Molly Clemens Molly Clemens Michele Faralli Jorge Lagreze Luana Bontempo Stefano Piazza Claudio Varotto Mickael Malnoy Walter Oechel Walter Oechel Annapaola Rizzoli Lorenza Dalla Costa VvEPFL9-1 Knock-Out via CRISPR/Cas9 Reduces Stomatal Density in Grapevine Frontiers in Plant Science Vitis vinifera stomata genome editing climate change water-use efficiency |
| title | VvEPFL9-1 Knock-Out via CRISPR/Cas9 Reduces Stomatal Density in Grapevine |
| title_full | VvEPFL9-1 Knock-Out via CRISPR/Cas9 Reduces Stomatal Density in Grapevine |
| title_fullStr | VvEPFL9-1 Knock-Out via CRISPR/Cas9 Reduces Stomatal Density in Grapevine |
| title_full_unstemmed | VvEPFL9-1 Knock-Out via CRISPR/Cas9 Reduces Stomatal Density in Grapevine |
| title_short | VvEPFL9-1 Knock-Out via CRISPR/Cas9 Reduces Stomatal Density in Grapevine |
| title_sort | vvepfl9 1 knock out via crispr cas9 reduces stomatal density in grapevine |
| topic | Vitis vinifera stomata genome editing climate change water-use efficiency |
| url | https://www.frontiersin.org/articles/10.3389/fpls.2022.878001/full |
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