Molecular Plant Breeding 2024, Vol.15, No.5, 209-219 http://genbreedpublisher.com/index.php/mpb 209 Review Article Open Access Application of Multi-Gene Stacking Strategies in Citrus Pest Resistance Breeding: From Theory to Practice Yiwei Li 1, Wenbin Dong2, Liyu Liang1, Fuping Liu1, Hongli Li 1, Huihong Liao1 , XiWang1 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 emails: liaohuihong2001@163.com; wangxi3951@126.com Molecular Plant Breeding, 2024, Vol.15, No.5 doi: 10.5376/mpb.2024.15.0021 Received: 07 Aug., 2024 Accepted: 10 Sep., 2024 Published: 18 Sep., 2024 Copyright © 2024 Li 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: Li Y.W., Dong W.B., Liang L.Y., Liu F.P., Li H.L., Liao H.H., and Wang X., 2024, Application of multi-gene stacking strategies in citrus pest resistance breeding: from theory to practice, Molecular Plant Breeding, 15(5): 209-219 (doi: 10.5376/mpb.2024.15.0021) Abstract Techniques such as gene aggregation, marker-assisted selection (MAS), and CRISPR-based genome editing have shown promising results in improving citrus resistance to pests and diseases. Gene polymerization has been successfully used to develop long-lasting resistant lines that are resistant to a variety of biological stresses, the combination of early-maturing transgenic plants with MAS has accelerated the breeding process and greatly reduced the time required to develop resistant varieties, and the CRISPR/Cas system has also played an important role in breeding citrus varieties that are resistant to various pathogens. The combination of advanced genetic engineering techniques and traditional breeding methods has the potential to produce robust citrus varieties that can withstand the challenges of multiple pests. Continued research and optimization of these strategies are essential for sustainable citrus production and improved crop resilience. This study highlights the significant advances in genetic engineering and biotechnology methods in the development of insect resistant citrus varieties, and aims to explore the application of polygenic stacking strategies in citrus pest resistance breeding, focusing on the theoretical basis and practical implementation of these strategies to improve the resistance of citrus to various pests. Keywords Multi-gene stacking; Citrus pest resistance; Genetic engineering; Marker-assisted selection; CRISPR/Cas systems 1 Introduction Citrus cultivation is a cornerstone of the global agricultural economy, with citrus fruits being among the most widely produced and consumed fruits worldwide. Citrus species, including oranges, lemons, limes, and grapefruits, are cultivated in over 130 countries, predominantly in tropical and subtropical regions (Sun et al., 2019). The citrus industry not only provides essential nutrients to millions of people but also supports the livelihoods of countless farmers and workers involved in its production, processing, and distribution. The economic significance of citrus is underscored by its substantial contribution to the agricultural GDP of many countries, making it a vital sector for both local and international markets (Poles et al., 2020). Despite its economic importance, citrus cultivation faces significant challenges, particularly from pests and diseases. Pests such as the Asian citrus psyllid, which spreads Huanglongbing (HLB), and pathogens causing citrus canker and citrus tristeza virus (CTV), have severely impacted citrus yields and fruit quality (Sendín and Filippone, 2018; Endo et al., 2020). These biotic stresses not only reduce the productivity of citrus orchards but also increase production costs due to the need for pest management and disease control measures. Traditional breeding methods to develop pest-resistant citrus varieties are often hampered by the long juvenility period, high heterozygosity, and complex reproductive biology of citrus species (Rauf et al., 2013; Sendín and Filippone, 2018). In response to these challenges, multi-gene stacking strategies have emerged as a promising approach to enhance pest resistance in citrus. This technique involves the simultaneous introduction of multiple resistance genes into a single plant, thereby providing broad-spectrum and durable resistance against a range of pests and diseases (Dormatey et al., 2020; Li et al., 2020). Advances in genetic engineering and molecular breeding techniques, such as marker-assisted selection (MAS) and genome editing, have facilitated the development of multi-gene stacking systems. These systems have been successfully applied in other crops, such as rice, to confer resistance to multiple pests and diseases, demonstrating their potential applicability in citrus breeding (Sun et al., 2019; Li et al., 2020).
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