Molecular Pathogens, 2025, Vol.16, No.6, 285-293 http://microbescipublisher.com/index.php/mp 291 8 Conclusion and Future Perspectives CRISPR/Cas9 technology has gained rapid popularity in plant disease resistance research in recent years, but its emergence was not sudden. In the past, people have been looking for a way to precisely and cost-effectively modify plant genes. CRISPR precisely meets these demands. For pathogens such as viruses, fungi and bacteria, the resistance of many plants has significantly improved after they have modified their genes with it. The principle is actually not complicated. It involves knocking out the virulence genes of the pathogen or the susceptibility genes of the plant itself, making it impossible for the disease to strike. Moreover, this technology can modify traits without the use of exogenous DNA and without involving genetic modification, which is particularly crucial for promoting green agriculture. However, things are not that simple. No matter how useful CRISPR is, there are still many "bottlenecks". For instance, the problem of off-target has not been completely resolved. Sometimes you think only one site has been moved, but in the end, the genes adjacent to it have also been affected. Moreover, the interaction between plants and pathogens is inherently complex, and the current understanding is far from sufficient, let alone how to efficiently apply these editing tools to various crops. Promotion is also not an easy task. There are many ambiguous areas in the regulatory framework, and public attitudes are inconsistent. The lack of clear supervision and reliable biosafety data has added many uncertainties to the implementation of this technology. What should I do next? Improving specificity is definitely the key point. After all, we can no longer let the editors be distracted. The delivery efficiency also needs to be improved; otherwise, no matter how good the laboratory effect is, it won't be able to be fully utilized in the fields. The development of multi-target editing holds great promise, especially for some crop environments where multiple pathogens coexist. For CRISPR to truly take root in agriculture, it may still need to be combined with precision agriculture. For instance, data can be used to monitor the dynamics of diseases, and then precise editing strategies can be designed. Only in this way can we deal with the constantly mutating new pathogens. Of course, policy coordination cannot be absent - clearer legal provisions are needed, and more people need to truly understand the ins and outs of this technology. Only when trust is established among technology, regulation and the public can CRISPR crops truly move to the fields and become part of daily life. Acknowledgements Thank you to all members of Hainan Biotechnology Research Institute for their help and support during this research. Conflict of Interest Disclosure The authors confirm that the study was conducted without any commercial or financial relationships and could be interpreted as a potential conflict of interest. References Ahmad S., Wei X., Sheng Z., Hu P., and Tang S., 2020, CRISPR/Cas9 for development of disease resistance in plants: recent progress limitations and future prospects, Briefings in Functional Genomics, 19(1): 26-39. https://doi.org/10.1093/bfgp/elz041 Ahmed A., Khan A., Negm M., Iqbal R., Azhar M., Khan S., and Rana I., 2024, Enhancing cotton resilience to challenging climates through genetic modifications, Journal of Cotton Research, 7(1): 196-206. https://doi.org/10.1186/s42397-024-00171-4 Allemailem K., 2024, Recent advances in understanding the molecular mechanisms of multidrug resistance and novel approaches of CRISPR/Cas9-based genome-editing to combat this health emergency, International Journal of Nanomedicine, 19: 1125-1143. https://doi.org/10.2147/ijn.s453566 Billah M., Li F., and Yang Z., 2021, Regulatory network of cotton genes in response to salt drought and wilt diseases (Verticillium and Fusarium): progress and perspective, Frontiers in Plant Science, 12: 759245. https://doi.org/10.3389/fpls.2021.759245 Binyameen B., Khan Z., Khan S., Ahmad A., Munawar N., Mubarik M., Riaz H., Ali Z., Khan A., Qusmani A., Abd-Elsalam K., and Qari S., 2021, Using multiplexed CRISPR/Cas9 for suppression of cotton leaf curl virus, International Journal of Molecular Sciences, 22(22): 12543. https://doi.org/10.3390/ijms222212543
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