International Journal of Horticulture, 2024, Vol.14, No.3, 142-155 http://hortherbpublisher.com/index.php/ijh 142 Research Advances Open Access Developing Citrus Germplasm Resistant to Asian Citrus Psyllid Using CRISPR/Cas9 Gene Editing Technology: Recent Advances and Challenges XiWang1, Yiwei Li 1, Fuping Liu1, Wenbin Dong2, Liyu Liang1, Dongkui Chen1, Hongli Li 1, Huihong Liao1 1 Horticultural Research Institute, Guangxi Academy of Agricultural Sciences, Nanning, 530007, Guangxi, China 2 Agriculture Resource and Environment Research Institute, Guangxi Academy of Agricultural Sciences, Nanning, 530007, Guangxi, China Corresponding author: liaohuihong2001@163.com International Journal of Horticulture, 2024, Vol.14, No.3 doi: 10.5376/ijh.2024.14.0016 Received: 02 May, 2024 Accepted: 03 Jun., 2024 Published: 15 Jun., 2024 Copyright © 2024 Wang et al., This is an open access article published under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Preferred citation for this article: Wang X., Li Y.W., Liu F.P., Dong W.B., Liang L.Y., Chen D.K., Li H.L., and Liao H.H., 2024, Developing citrus germplasm resistant to Asian citrus psyllid using CRISPR/Cas9 gene editing technology: recent advances and challenges, International Journal of Horticulture, 14(3): 142-155 (doi: 10.5376/ijh.2024.14.0016) Abstract This study aims to explore recent advances and challenges in developing citrus germplasm resistant to the Asian citrus psyllid (ACP) using CRISPR/Cas9 gene editing technology. The focus is on identifying key genetic targets, evaluating the effectiveness of CRISPR/Cas9-mediated edits, and discussing the implications for sustainable citrus production. Several case studies demonstrate the potential of CRISPR/Cas9 to enhance resistance without compromising yield and fruit quality. Advances in CRISPR/Cas9 techniques, such as base and prime editing, have improved the precision and efficiency of gene editing in citrus. Additionally, field trials have validated the effectiveness of these edited plants in real-world conditions. The findings underscore the significant potential of CRISPR/Cas9 technology in developing ACP-resistant citrus germplasm. However, technical challenges, off-target effects, genetic stability, and regulatory and public acceptance issues remain. Continued research, interdisciplinary collaboration, and clear regulatory frameworks are essential to fully realize the benefits of CRISPR/Cas9 in citrus breeding. These efforts are crucial for ensuring the long-term sustainability and resilience of the citrus industry. Keywords Citrus germplasm; Asian citrus psyllid (ACP); CRISPR/Cas9; Huanglongbing (HLB); Gene editing; Plant resistance; Sustainable agriculture The Asian citrus psyllid (ACP), scientifically known as Diaphorina citri, is a small sap-sucking insect native to Asia but has now spread to many citrus-growing regions worldwide, including the Americas (Chen et al., 2021; Carlson et al., 2022). It poses a significant threat to citrus crops globally. ACP is the primary vector for the bacteriumCandidatus Liberibacter asiaticus, which causes Huanglongbing (HLB), also known as citrus greening disease (Hall et al., 2013). This disease has caused widespread devastation in citrus groves worldwide, with infected trees showing symptoms such as yellowing leaves, deformed and bitter-tasting fruit, premature fruit drop, and ultimately tree death, leading to substantial economic losses (El-Shesheny et al., 2013; Zhang et al., 2020). The rapid spread of ACP and the persistence of HLB make conventional pest management strategies, such as chemical control and biological agents, less effective. Therefore, the citrus industry urgently needs sustainable and long-term solutions to combat this pest and its associated disease. CRISPR/Cas9 gene editing technology has emerged as a revolutionary tool in genetic research and plant breeding (Ahmad et al., 2020). Derived from a bacterial immune system, CRISPR/Cas9 enables precise modification of DNA sequences within organisms. This technology offers several advantages over traditional breeding and genetic modification methods, including high specificity, efficiency, and the ability to introduce targeted changes without leaving foreign DNA in the host genome. In plant breeding, CRISPR/Cas9 holds immense potential for developing crops with enhanced traits such as disease resistance, improved yield, and stress tolerance (Chaverra-Rodriguez et al., 2023). Given its precision and versatility, CRISPR/Cas9 presents a promising approach to addressing the challenges posed by ACP and HLB in citrus crops (Carlson et al., 2022; Chaverra-Rodriguez et al., 2023).
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