Molecular Plant Breeding 2024, Vol.15, No.5, 209-219 http://genbreedpublisher.com/index.php/mpb 217 modified citrus varieties (Conti et al., 2021). Ultimately, the goal is to achieve a sustainable and resilient citrus industry that can thrive in the face of emerging pest and disease threats. 9 Concluding Remarks The application of multi-gene stacking strategies in citrus pest resistance breeding has shown significant promise in enhancing disease resistance and improving agronomic traits. Multi-gene stacking, or transgene stacking, involves the introduction of multiple genes into a plant to confer resistance to various pests and diseases. This approach has been more effective than single-gene technology in providing durable resistance and improving crop yield. In citrus, genetic engineering techniques such as CRISPR/Cas9 have been successfully employed to develop disease-resistant varieties, particularly against bacterial canker and huanglongbing. The use of new plant breeding techniques (NPBTs) has also been highlighted as a crucial method for overcoming traditional breeding limitations and achieving improved resistance to biotic and abiotic stresses. For researchers, the findings underscore the importance of continuing to explore and refine multi-gene stacking techniques. The development of efficient transgene stacking systems and the optimization of protocols for plant regeneration are critical areas for future research. Breeders can leverage these advanced genetic engineering tools to develop citrus varieties with enhanced resistance to multiple pests and diseases, thereby reducing the reliance on chemical pesticides and improving crop yield and quality. Policymakers should consider the regulatory frameworks surrounding the use of genetically modified organisms (GMOs) and NPBTs. Clear guidelines and support for the adoption of these technologies can facilitate their integration into agricultural practices, ultimately contributing to sustainable agriculture and food security. Further research is needed to address the challenges associated with multi-gene stacking, such as the potential for evolved resistance in pests and the need for marker-free systems. Interdisciplinary collaboration between geneticists, plant pathologists, agronomists, and policymakers is essential to advance the development and implementation of these technologies. By working together, these stakeholders can ensure that the benefits of multi-gene stacking strategies are fully realized, leading to more resilient and productive citrus crops. Additionally, exploring the integration of conventional breeding with genetic engineering approaches can provide a comprehensive strategy for achieving durable resistance in citrus. Acknowledgments The authors would like to thank Dr. X. Fang, General Director of the Hainan Institute of Tropical Agricultural Resources, for reading the manuscript and providing valuable suggestions for revision. Additionally, thanks are extended to the two anonymous peer reviewers for their rigorous review and constructive comments on the paper. Funding This research was supported by the Nanning Major Science and Technology Project (20232080), the Guangxi Key Research and Development Program (Guike AB18294015), and the Basic Scientific Research Project of Guangxi Academy of Agricultural Sciences (2015YM03). Conflict of Interest Disclosure The authors affirm that this research was conducted without any commercial or financial relationships that could be construed as a potential conflict of interest. References Ainley W., Sastry-Dent L., Welter M., Murray M., Zeitler B., Amora R., Corbin D., Miles R., Arnold N., Strange T., Simpson M., Cao Z., Carroll C., Pawelczak K., Blue R., West K., Rowland L., Perkins D., Samuel P., Dewes C., Shen L., Sriram S., Evans S., Rebar E., Zhang L., Gregory P., Urnov F., Webb S., and Petolino J., 2013, Trait stacking via targeted genome editing, Plant Biotechnology Journal, 11(9): 1126-1134. https://doi.org/10.1111/pbi.12107 PMid:23953646
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