Triticeae Genomics and Genetics, 2024, Vol.15, No.2, 77-87 http://cropscipublisher.com/index.php/lgg 84 Fiture 3 CRISPR/Cas9-based editing of an endogenous ubiquitin gene in wheat microspores (Adopted from Bhowmik et al., 2018) Image caption: (A) Schematic diagram of the pEEE005-NLSCas9/gUbi1 and pEEE006-NLSCas9/gUbi1/GFP constructs used for mutagenesis of in wheat microspores; (B) PCR/restriction enzyme assay to detect mutations in TaUbiL1induced by Cas9/ gUbi1; (C) Deletions detected at the cleavage site in sequenced products of all three homoeologs of TaUbiL1 from wheat microspores transfected with pEEE005-NLSCas9/gUbi1 or pEEE006-NLSCas9/gUbi1/GFP constructs; (D) Sanger sequencing electropherograms showing mutations inTaUbiL1homoeologues from the A, B and D genomes of wheat (Adopted from Bhowmik et al., 2018) 7.2 Impact on food production and quality The reduction of gluten proteins in wheat through breeding strategies has significant implications for food production and quality. Gluten proteins are crucial for the viscoelastic properties of dough, which affect the texture and quality of bread and other wheat-based products. While reducing gliadins can lower the allergenic potential of wheat, it is essential to balance this with the need to maintain or improve the functional properties of wheat flour. Studies have shown that cultivated barley, which has lower immunogenic epitopes and gluten content, could serve as an alternative cereal with favorable properties for food production (Ozuna and Barro, 2018). Furthermore, the diverse allelic pool of wild wheat relatives, such as T. urartu, offers opportunities to enhance wheat quality traits, including flour quality and carotenoid concentrations, which can contribute to the development of high-quality, allergen-free wheat products (Talini et al., 2019). 7.3 Regulatory and safety considerations The development and commercialization of allergen-free wheat varieties must adhere to stringent regulatory and safety standards to ensure consumer safety. Regulatory agencies require comprehensive testing to confirm the absence or significant reduction of immunogenic epitopes in new wheat varieties. This includes molecular characterization, immunological assays, and clinical trials to assess the safety and efficacy of the allergen-free wheat. The variability in the number of CD epitopes among different Triticeae species and genomes highlights the need for precise and thorough evaluation of each new variety (Ozuna and Barro, 2018). Additionally, regulatory frameworks must address labeling requirements to inform consumers about the allergen-free status of wheat products, ensuring transparency and trust in the marketplace.
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