Molecular Pathogens, 2025, Vol.16, No.6, 285-293 http://microbescipublisher.com/index.php/mp 285 Research Insight Open Access Restoration of Cotton Disease Resistance by CRISPR-Mediated Disruption of Key Genes in Drug-Resistant Pathogens Shujuan Wang, Jiong Fu Hainan Provincial Key Laboratory of Crop Molecular Breeding, Sanya, 572025, Hainan, China Corresponding author: jiong.fu@hitar.org Molecular Pathogens, 2025, Vol.16, No.6 doi: 10.5376/mp.2025.16.0029 Received: 17 Oct., 2025 Accepted: 27 Nov., 2025 Published: 16 Dec., 2025 Copyright © 2025 Wang and Fu, 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 S.J., and Fu J., 2025, Restoration of cotton disease resistance by CRISPR-mediated disruption of key genes in drug-resistant pathogens, Molecular Pathogens, 16(6): 285-293 (doi: 10.5376/mp.2025.16.0029) Abstract The widespread emergence of drug-resistant pathogenic bacteria poses a severe challenge to the sustainable production of cotton. The effectiveness of traditional chemical control measures is gradually weakening, and it is urgent to develop new and precise control strategies. In this study, CRISPR/Cas genome editing technology was employed to target and interfere with the key functional genes in the main drug-resistant pathogenic bacteria of cotton (such as Verticillium dahliae and Fusarium oxysporum). By identifying the key genes related to pathogenicity and drug resistance, Experiments have confirmed that CRISPR interference can significantly weaken the pathogenic ability of pathogenic bacteria, and the recovery effect of this strategy on the disease resistance of cotton has been evaluated. The results show that CRISPR interference treatment can activate the host immune response and improve field performance. Through the case analysis of VdSOD1 and FOW1 gene knockout, this study further explored the effectiveness of pathogen gene editing in reducing pathogenicity and indirectly restoring the disease resistance of cotton. This research provides a theoretical basis and application path for pathogen-targeted green prevention and control, and is expected to promote its development in precision agriculture. Keywords CRISPR/Cas system; Cotton disease resistance; Drug-resistant pathogenic bacteria; Interference of target genes; Green prevention and control strategy 1 Introduction Whether cotton is grown well or not, apart from the variety and management, disease problems are often the key to determining the yield and quality. In major production areas, pathogens such as viruses, fungi and bacteria have never been less troublesome for cotton. Sometimes, once the disease breaks out, not only is the yield reduced severely, but the quality of the fibers also deteriorates. The losses of farmers can be imagined. For instance, the cotton leaf roll disease (CLCuD) transmitted by bedlice has always been a headache. Also, fungal pathogens like the Trichoderma lucidum have put considerable pressure on the production areas (Khan et al., 2020; Mubarik et al., 2021; Zhu et al., 2022). In addition to the direct reduction in production, these diseases have also increased the cost of field management, reduced the market value of products, and affected the entire cotton industry chain (Bukhari et al., 2021). Traditional prevention and control measures, which relied on chemical agents and conventional breeding, are becoming less and less effective at present. One of the reasons is that pathogens mutate so rapidly that some can even acquire drug resistance rapidly through horizontal gene transfer (Moniruzzaman et al., 2020). The drugs or disease-resistant varieties that might be effective at first will soon be "cracked". Sometimes, before a harvest is over, new drug-resistant strains have already emerged. This situation of "chasing each other" makes the prevention and control work more difficult and also reminds us that we can no longer rely solely on the old methods. New ideas are needed now more than ever for persistent resistance strategies that can truly "outpace" pathogen changes (Ahmad et al., 2020). This study explored the use of CRISPR-mediated genome editing technology to destroy the key genes of drug-resistant cotton pathogens, identify the key pathogen genes causing drug resistance, target these genes with CRISPR/Cas9 technology, and evaluate their impact on the disease resistance of cotton. Through precise genome
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