Enhancing tolerance to Phytophthora spp. in eggplant through DMR6–1 CRISPR/Cas9 knockout

Agricultural production is affected by the worsening effects of climate change with severe yield losses caused by rising temperatures, water scarcity and consequent modifications in the interactions between crops, pests, and pathogens. The availability of stress-tolerant plants will thus be a key po...

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Main Authors: Martina Ferrero, Danila Valentino, Anna Maria Milani, Cinzia Comino, Sergio Lanteri, Alberto Acquadro, Andrea Moglia
Format: Article
Language:English
Published: Elsevier 2024-12-01
Series:Plant Stress
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Online Access:http://www.sciencedirect.com/science/article/pii/S2667064X24003440
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author Martina Ferrero
Danila Valentino
Anna Maria Milani
Cinzia Comino
Sergio Lanteri
Alberto Acquadro
Andrea Moglia
author_facet Martina Ferrero
Danila Valentino
Anna Maria Milani
Cinzia Comino
Sergio Lanteri
Alberto Acquadro
Andrea Moglia
author_sort Martina Ferrero
collection DOAJ
description Agricultural production is affected by the worsening effects of climate change with severe yield losses caused by rising temperatures, water scarcity and consequent modifications in the interactions between crops, pests, and pathogens. The availability of stress-tolerant plants will thus be a key point to guarantee the world food security in the next future. To this purpose, a significant contribution might be provided by the New Genomic Techniques (NGTs), such as CRISPR/Cas9, which allow to insert targeted modifications in the plants’ genomes offering new opportunities for crop improvement. Susceptibility genes encode proteins that pathogens can take advantage of during the colonization process, and their disabling confers a broad-spectrum and long-lasting pathogen tolerance to the plant. Among the S-genes, Downy Mildew Resistance 6 (DMR6) encodes an enzyme involved in Salicylic Acid (SA) degradation, and its inactivation in other Solanaceae species has proven to increase SA levels and confer tolerance to a broad spectrum of pathogens. We identified two orthologs of this gene in eggplant's genome, namely SmDMR6–1 and SmDMR6–2. In the ‘Black Beauty’ cultivar, only SmDMR6–1 expression significantly increased upon infection by the two oomycetes Phytophthora infestans and Phytophthora capsici, suggesting its involvement in the regulation of plant responses to biotic stresses. Here we report, for the first time in eggplant, the knockout of SmDMR6–1 gene through CRISPR/Cas9 technology. The regenerated T0 plants were screened by Sanger sequencing and one was selected and self-pollinated to generate T1 and then T2 plants. The mutant lines were subjected to pathogen assays which highlighted an increased tolerance to infection by P. infestans and P. capsici, if compared to non-edited plants.
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language English
publishDate 2024-12-01
publisher Elsevier
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series Plant Stress
spelling doaj-art-85d88e2234c245509d05dc04ce920f4f2024-12-19T11:02:14ZengElsevierPlant Stress2667-064X2024-12-0114100691Enhancing tolerance to Phytophthora spp. in eggplant through DMR6–1 CRISPR/Cas9 knockoutMartina Ferrero0Danila Valentino1Anna Maria Milani2Cinzia Comino3Sergio Lanteri4Alberto Acquadro5Andrea Moglia6DISAFA, Department of Agricultural, Forest and Food Sciences, University of Turin, 10095, Grugliasco, TO, ItalyDISAFA, Department of Agricultural, Forest and Food Sciences, University of Turin, 10095, Grugliasco, TO, ItalyDISAFA, Department of Agricultural, Forest and Food Sciences, University of Turin, 10095, Grugliasco, TO, ItalyDISAFA, Department of Agricultural, Forest and Food Sciences, University of Turin, 10095, Grugliasco, TO, ItalyDISAFA, Department of Agricultural, Forest and Food Sciences, University of Turin, 10095, Grugliasco, TO, ItalyDISAFA, Department of Agricultural, Forest and Food Sciences, University of Turin, 10095, Grugliasco, TO, ItalyCorresponding author.; DISAFA, Department of Agricultural, Forest and Food Sciences, University of Turin, 10095, Grugliasco, TO, ItalyAgricultural production is affected by the worsening effects of climate change with severe yield losses caused by rising temperatures, water scarcity and consequent modifications in the interactions between crops, pests, and pathogens. The availability of stress-tolerant plants will thus be a key point to guarantee the world food security in the next future. To this purpose, a significant contribution might be provided by the New Genomic Techniques (NGTs), such as CRISPR/Cas9, which allow to insert targeted modifications in the plants’ genomes offering new opportunities for crop improvement. Susceptibility genes encode proteins that pathogens can take advantage of during the colonization process, and their disabling confers a broad-spectrum and long-lasting pathogen tolerance to the plant. Among the S-genes, Downy Mildew Resistance 6 (DMR6) encodes an enzyme involved in Salicylic Acid (SA) degradation, and its inactivation in other Solanaceae species has proven to increase SA levels and confer tolerance to a broad spectrum of pathogens. We identified two orthologs of this gene in eggplant's genome, namely SmDMR6–1 and SmDMR6–2. In the ‘Black Beauty’ cultivar, only SmDMR6–1 expression significantly increased upon infection by the two oomycetes Phytophthora infestans and Phytophthora capsici, suggesting its involvement in the regulation of plant responses to biotic stresses. Here we report, for the first time in eggplant, the knockout of SmDMR6–1 gene through CRISPR/Cas9 technology. The regenerated T0 plants were screened by Sanger sequencing and one was selected and self-pollinated to generate T1 and then T2 plants. The mutant lines were subjected to pathogen assays which highlighted an increased tolerance to infection by P. infestans and P. capsici, if compared to non-edited plants.http://www.sciencedirect.com/science/article/pii/S2667064X24003440Susceptibility genesPhytophthoraCRISPR/Cas9pathogen toleranceSolanum melongena L
spellingShingle Martina Ferrero
Danila Valentino
Anna Maria Milani
Cinzia Comino
Sergio Lanteri
Alberto Acquadro
Andrea Moglia
Enhancing tolerance to Phytophthora spp. in eggplant through DMR6–1 CRISPR/Cas9 knockout
Plant Stress
Susceptibility genes
Phytophthora
CRISPR/Cas9
pathogen tolerance
Solanum melongena L
title Enhancing tolerance to Phytophthora spp. in eggplant through DMR6–1 CRISPR/Cas9 knockout
title_full Enhancing tolerance to Phytophthora spp. in eggplant through DMR6–1 CRISPR/Cas9 knockout
title_fullStr Enhancing tolerance to Phytophthora spp. in eggplant through DMR6–1 CRISPR/Cas9 knockout
title_full_unstemmed Enhancing tolerance to Phytophthora spp. in eggplant through DMR6–1 CRISPR/Cas9 knockout
title_short Enhancing tolerance to Phytophthora spp. in eggplant through DMR6–1 CRISPR/Cas9 knockout
title_sort enhancing tolerance to phytophthora spp in eggplant through dmr6 1 crispr cas9 knockout
topic Susceptibility genes
Phytophthora
CRISPR/Cas9
pathogen tolerance
Solanum melongena L
url http://www.sciencedirect.com/science/article/pii/S2667064X24003440
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