Plant Gene and Traits 2024, Vol.15, No.4, 195-206 http://genbreedpublisher.com/index.php/pgt 200 resistance against leaf-chewing insects. Mutations in this gene resulted in increased flavonoid biosynthesis, which is associated with insect resistance (Zhang et al., 2022). Similarly, genes involved in the plant's defense mechanisms against fungal and bacterial pathogens have been modified to improve resistance (Borrelli et al., 2018; Erdoğan et al., 2023). 7.3 Case studies of successful genetic engineering efforts for resistance traits in legumes A typical case study highlights the successful application of genetic engineering and CRISPR-Cas9 in developing pest and disease-resistant legumes. Using CRISPR-Cas9, researchers introduced targeted mutations in the GmUGT gene, resulting in soybean varieties with enhanced resistance to pests like Helicoverpa armigera and Spodoptera litura. This modification did not affect the overall plant phenotype, demonstrating the precision and effectiveness of CRISPR-Cas9 in pest resistance breeding (Figure 2) (Zhang et al., 2022). This case study underscore the transformative impact of CRISPR-Cas9 technology in resistance breeding, offering new avenues for developing robust legume varieties capable of withstanding biotic stresses. Figure 2 The CRISPR/Cas9-mediated mutagenesis of GmUGTenhanced resistance to H. armigera and S. litura in soybean (Adopted from Zhang et al., 2022) Image caption: (A) The phenotype of detached leaves of ko-3, ko-5, and WT plants attacked by H. armigera for 3 days (bar = 2 cm). (B) Percentage of leaf area loss in ko-3, ko-5, and WT plants attacked by H. armigera for 3 days (n = 50 biological repeats). (C) The phenotype of H. armigera larvae fed detached leaves of ko-3, ko-5, and WT plants for 7 days (bar = 0.2 cm). (D) The average weight of H. armigera larvae that were fed detached leaves of ko-3, ko-5, and WT plants for 7 days (n = 50 larvae). (E) The phenotype of detached leaves of ko-3, ko-5, and WT that was attacked by S. litura for 3 days (bar = 2 cm). (F) Percentage of leaf area loss in ko-3, ko-5, and WT plants attacked by S. litura for 3 days (n = 50 biological repeats). (G) The phenotype of S. litura larvae that were fed detached leaves of ko-3, ko-5, and WT plants for 7 days (bar = 0.2 cm). (H) The average weight of S. litura larvae that were fed detached leaves of ko-3, ko-5, and WT for 7 days (n = 50 larvae). Data shown are means and standard deviations. Statistically significant differences are marked with asterisks (**p< 0.01; Student’s t-test) (Adopted from Zhang et al., 2022) 8 Case Study: Molecular Breeding for Resistance to Fusarium Wilt in Chickpea 8.1 Overview of Fusarium wilt and its impact on chickpea production Fusarium wilt, caused by the soil-borne fungus Fusarium oxysporum f. sp. Ciceris (FOC), is a major disease affecting chickpea (Cicer arietinum L.) production globally. This disease can lead to significant yield losses, sometimes up to 100% under severe conditions (Jha et al., 2020; Choudhary et al., 2022). Fusarium wilt is particularly problematic because it affects the vascular system of the plant, leading to wilting and eventual plant death. The disease is exacerbated by certain environmental conditions and is a persistent problem in many chickpea-growing regions (Mannur et al., 2018; Choudhary et al., 2022).
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