Legume Genomics and Genetics 2024, Vol.15, No.4, 199-209 http://cropscipublisher.com/index.php/lgg 200 breeding methods. It enables precise and targeted modifications, reducing the time and effort required to develop improved crop varieties. Unlike conventional breeding, which relies on random mutagenesis or genetic recombination, CRISPR/Cas9 allows for the direct manipulation of specific genes associated with desirable traits, such as stress resilience and nutritional quality. Additionally, the technology has been successfully applied to various legume crops, including soybean, cowpea, and chickpea, demonstrating its potential to overcome the limitations of traditional breeding. However, regulatory and public acceptance challenges remain, which need to be addressed to fully realize the benefits of CRISPR/Cas9 in legume breeding. The study is to explore the potential of CRISPR/Cas9 genome editing in legume crops for functional genomics and breeding. By leveraging recent advances in genomic resources and gene-editing technologies, this study aims to address the current challenges in legume breeding and enhance the genetic gain related to yield, stress resilience, and nutritional quality. The study will also examine the regulatory landscape and public acceptance of CRISPR/Cas9 technology to ensure its successful implementation in legume crop improvement. 2 Functional Genomics in Legumes 2.1 Role of functional genomics in crop improvement Functional genomics plays a crucial role in understanding the specific roles of genes in legume crops. By utilizing advanced genome-editing technologies such as CRISPR/Cas9, researchers can precisely target and modify genes to study their functions. This approach has been successfully applied in model legumes like Medicago truncatula and crop legumes such as soybean, enabling the identification of genes responsible for various traits (Bhowmik et al., 2021). The simplicity and efficiency of CRISPR/Cas9 make it a standout choice for targeted genome editing, facilitating the investigation of gene functions and the development of valuable traits in legumes (Meng et al., 2016). Functional genomics also aids in the identification of key traits that are essential for crop improvement. Traits such as yield, stress resilience, and nutritional quality are of particular interest. Recent advancements in genomic resources for legumes have laid a solid foundation for the application of transformative breeding technologies, including CRISPR/Cas9, to enhance these traits (Bhowmik et al., 2021). For instance, the CRISPR/Cas9 system has been used to target genes involved in fatty acid composition in soybean, resulting in improved nutritional profiles (Do et al., 2019). 2.2 CRISPR/Cas9 as a tool for functional genomics CRISPR/Cas9 has revolutionized gene knockout studies in legumes by enabling precise and efficient gene disruption. This technology has been employed to create knockouts in various legume species, including soybean and cowpea, to study gene functions and improve traits (Ji et al., 2019; Bhowmik et al., 2021). For example, targeted mutagenesis using CRISPR/Cas9 in Medicago truncatula has facilitated the investigation of gene functions related to forage quality and yield (Meng et al., 2016). Beyond gene knockouts, CRISPR/Cas9 can also be used for gene activation and repression. This is achieved by fusing the Cas9 protein with transcriptional activators or repressors, allowing for the upregulation or downregulation of target genes. Such manipulations can help in understanding gene regulatory networks and their impact on important agronomic traits (Arora and Narula, 2017; Thomson et al., 2019). The ability to modulate gene expression using CRISPR/Cas9 provides a powerful tool for functional genomics studies in legumes. 2.3 Case study: functional genomics of drought tolerance in soybeans Drought tolerance is a critical trait for soybean cultivation, especially in regions prone to water scarcity. Functional genomics approaches, including transcriptomics and CRISPR/Cas9, have been employed to identify genes responsive to drought stress. These studies have revealed several candidate genes that play significant roles in drought tolerance (Figure 1) (Arora and Narula, 2017; Cai et al., 2020). Understanding the functions of these genes is essential for developing drought-resistant soybean varieties.
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