Bioscience Methods 2024, Vol.15, No.6, 315-326 http://bioscipublisher.com/index.php/bm 320 5.2 Research outcomes on yield improvement through the editing of regulatory genes Recent research has demonstrated the effectiveness of CRISPR/Cas9 in improving wheat yield by editing regulatory genes. For example, the targeted mutagenesis of the TaARE1 gene in wheat has led to increased nitrogen use efficiency and delayed senescence, resulting in higher grain yield (Zhang et al., 2021a). This study highlights the potential of CRISPR/Cas9 to enhance yield-related traits by manipulating key regulatory genes involved in nutrient utilization and plant development. Another significant outcome is the editing of promoter regions of CLEgenes, which are involved in the regulation of meristem size and, consequently, yield-related traits. By creating weak promoter alleles and null alleles of these genes, researchers have successfully increased grain yield in maize, providing a promising approach for similar improvements in wheat (Liu et al., 2021b). These findings underscore the potential of CRISPR/Cas9 to fine-tune gene expression and achieve desired agronomic traits. 5.3 Case studies on enhancing wheat grain quality (e.g., protein content, gluten strength) via editing CRISPR/Cas9 technology has also been employed to enhance wheat grain quality, focusing on traits such as protein content and gluten strength. In one study, researchers targeted four genes related to grain quality: pinb, waxy, ppo, and psy. These genes are involved in determining wheat grain hardness, starch quality, and dough color. The precise editing of these genes resulted in wheat varieties with improved grain quality attributes, demonstrating the versatility of CRISPR/Cas9 in modulating complex traits (Zhang et al., 2021). Another case study involved the use of CRISPR/Cas9 to improve the nutritional components of wheat. By targeting specific genes that influence protein content and gluten strength, researchers have been able to develop wheat varieties with enhanced nutritional profiles. This approach not only improves the quality of wheat but also addresses the growing demand for high-quality, nutritious food (Liu et al., 2021a; 2022). The ability to make precise modifications to the wheat genome opens up new possibilities for enhancing grain quality and meeting consumer preferences. 6 Improvement of Wheat Nutritional Content 6.1 The demand for improving wheat nutritional value (e.g., micronutrients, reduction of anti-nutritional factors) The global demand for wheat with enhanced nutritional value is driven by the need to address malnutrition and improve public health. Wheat is a staple food for a significant portion of the world's population, and its nutritional enhancement can have a profound impact on human health. Traditional breeding methods have been employed to improve wheat's nutritional profile, but these methods are often time-consuming and less precise. The advent of CRISPR/Cas9 technology offers a promising alternative, enabling precise modifications to enhance the nutritional content of wheat (Arora and Narula, 2017; Eş et al., 2019; Liu et al., 2021). Micronutrient deficiencies, such as those of iron, zinc, and vitamins, are prevalent in many parts of the world. Enhancing the micronutrient content of wheat can help alleviate these deficiencies. Additionally, reducing anti-nutritional factors, such as phytic acid, which inhibits the absorption of essential minerals, is crucial for improving the bioavailability of nutrients in wheat. CRISPR/Cas9 technology allows for targeted editing of genes involved in nutrient biosynthesis and anti-nutritional factor production, making it a powerful tool for nutritional improvement (Chen et al., 2019; Li et al., 2021; Wang et al., 2021). 6.2 Research examples of enhancing nutritional content through CRISPR/Cas9 Several studies have demonstrated the successful application of CRISPR/Cas9 in enhancing the nutritional content of wheat. For instance, researchers have used CRISPR/Cas9 to target and edit genes involved in the biosynthesis of essential nutrients. One notable example is the editing of the TaABCC6 gene, which has been shown to increase the bioavailability of zinc and iron in wheat grains by reducing the levels of phytic acid, an anti-nutritional factor (Cui et al., 2019; Wang et al., 2021a).
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