Assessing drought stress response in low-gliadin wheat developed via RNAi and CRISPR/Cas

The gluten proteins are responsible for the visco-elastic properties of flour, but they also trigger the immune response in celiac disease patients. For that reason, low-gliadin RNA interference (RNAi) and CRISPR bread and durum wheat lines were generated in previous works. This study evaluated thei...

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Main Authors: Latifa Chaouachi, María H. Guzmán-López, Chahine Karmous, Francisco Barro, Miriam Marín-Sanz
Format: Article
Language:English
Published: Elsevier 2025-06-01
Series:Plant Stress
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Online Access:http://www.sciencedirect.com/science/article/pii/S2667064X25001745
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author Latifa Chaouachi
María H. Guzmán-López
Chahine Karmous
Francisco Barro
Miriam Marín-Sanz
author_facet Latifa Chaouachi
María H. Guzmán-López
Chahine Karmous
Francisco Barro
Miriam Marín-Sanz
author_sort Latifa Chaouachi
collection DOAJ
description The gluten proteins are responsible for the visco-elastic properties of flour, but they also trigger the immune response in celiac disease patients. For that reason, low-gliadin RNA interference (RNAi) and CRISPR bread and durum wheat lines were generated in previous works. This study evaluated their drought stress response during the post-anthesis stage through gene expression and protein analyses. A drought-sensible durum wheat variety and the drought-tolerant landrace Aoujia were compared to the RNAi and CRISPR/Cas lines under the same conditions. The severe water stress treatment consisted of 25% field capacity (FC) applied three days after anthesis (DAA), while the 100% FC was set as control conditions. The expression levels of genes encoding enzymes (CAT and GPX), an enzyme involved in proline biosynthesis (P5CR), and galactinol synthase gene (GolS1) were assessed in response to drought stress. All the lines showed an increase of GPX, GolS1 and P5CR expression under drought. Particularly, Aouija and the D793 RNAi line had the greatest CAT, P5CR, and GolS1 expression compared to the others. Notably, some low-gliadin lines showed an overexpression of drought-related genes even under control conditions, possibly due to pleiotropic effects on other genes. The low-gliadin lines exhibited responses comparable to, or better than, their wild relatives, indicating an unaltered or improved stress response. Regarding the prolamins accumulation, RNAi and CRISPR lines retained low-gliadin content under severe drought, contrary to the wild-type lines, which increased their gliadin content drastically. This indicated the strong stability of the low-gliadin nature of these lines.
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spelling doaj-art-089b6e849f554192b5d940773b7fb73d2025-08-20T03:19:53ZengElsevierPlant Stress2667-064X2025-06-011610090610.1016/j.stress.2025.100906Assessing drought stress response in low-gliadin wheat developed via RNAi and CRISPR/CasLatifa Chaouachi0María H. Guzmán-López1Chahine Karmous2Francisco Barro3Miriam Marín-Sanz4Laboratory of Genetics and Cereal Breeding (LR14 AGR01), National Institute of Agronomy of Tunisia, Carthage University, 1082 Tunis, TunisiaDepartment of Plant Breeding, Institute for Sustainable Agriculture-Spanish National Research Council (IAS-CSIC), 14004 Córdoba, SpainLaboratory of Genetics and Cereal Breeding (LR14 AGR01), National Institute of Agronomy of Tunisia, Carthage University, 1082 Tunis, TunisiaDepartment of Plant Breeding, Institute for Sustainable Agriculture-Spanish National Research Council (IAS-CSIC), 14004 Córdoba, SpainDepartment of Plant Breeding, Institute for Sustainable Agriculture-Spanish National Research Council (IAS-CSIC), 14004 Córdoba, Spain; Corresponding author.The gluten proteins are responsible for the visco-elastic properties of flour, but they also trigger the immune response in celiac disease patients. For that reason, low-gliadin RNA interference (RNAi) and CRISPR bread and durum wheat lines were generated in previous works. This study evaluated their drought stress response during the post-anthesis stage through gene expression and protein analyses. A drought-sensible durum wheat variety and the drought-tolerant landrace Aoujia were compared to the RNAi and CRISPR/Cas lines under the same conditions. The severe water stress treatment consisted of 25% field capacity (FC) applied three days after anthesis (DAA), while the 100% FC was set as control conditions. The expression levels of genes encoding enzymes (CAT and GPX), an enzyme involved in proline biosynthesis (P5CR), and galactinol synthase gene (GolS1) were assessed in response to drought stress. All the lines showed an increase of GPX, GolS1 and P5CR expression under drought. Particularly, Aouija and the D793 RNAi line had the greatest CAT, P5CR, and GolS1 expression compared to the others. Notably, some low-gliadin lines showed an overexpression of drought-related genes even under control conditions, possibly due to pleiotropic effects on other genes. The low-gliadin lines exhibited responses comparable to, or better than, their wild relatives, indicating an unaltered or improved stress response. Regarding the prolamins accumulation, RNAi and CRISPR lines retained low-gliadin content under severe drought, contrary to the wild-type lines, which increased their gliadin content drastically. This indicated the strong stability of the low-gliadin nature of these lines.http://www.sciencedirect.com/science/article/pii/S2667064X25001745DroughtRNAiCRISPR/CasWheatLow-gliadin
spellingShingle Latifa Chaouachi
María H. Guzmán-López
Chahine Karmous
Francisco Barro
Miriam Marín-Sanz
Assessing drought stress response in low-gliadin wheat developed via RNAi and CRISPR/Cas
Plant Stress
Drought
RNAi
CRISPR/Cas
Wheat
Low-gliadin
title Assessing drought stress response in low-gliadin wheat developed via RNAi and CRISPR/Cas
title_full Assessing drought stress response in low-gliadin wheat developed via RNAi and CRISPR/Cas
title_fullStr Assessing drought stress response in low-gliadin wheat developed via RNAi and CRISPR/Cas
title_full_unstemmed Assessing drought stress response in low-gliadin wheat developed via RNAi and CRISPR/Cas
title_short Assessing drought stress response in low-gliadin wheat developed via RNAi and CRISPR/Cas
title_sort assessing drought stress response in low gliadin wheat developed via rnai and crispr cas
topic Drought
RNAi
CRISPR/Cas
Wheat
Low-gliadin
url http://www.sciencedirect.com/science/article/pii/S2667064X25001745
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