Molecular Pathogens, 2025, Vol.16, No.6, 285-293 http://microbescipublisher.com/index.php/mp 290 6.2 Functional validation of FOW1 gene disruption in Fusarium and its effect on virulence It's not only the Polyporus umbellatus that is a problem for cotton. Pathogens like Fusarium acuminatum also put considerable pressure on cotton. Some past genetic studies have mentioned that genes like FOW1 play a rather crucial role in Fusarium. If one wants to weaken the pathogenicity of this pathogen, knocking out FOW1 is a direct and effective approach. Experiments also verified this point. Strains with this gene knocked out exhibited a lower infectivity. Although it is a single gene, it involves multiple pathways such as hormone signaling and secondary metabolism (Billah et al., 2021). Once such genes are identified, the intervention of CRISPR becomes an important means of precise strikes. 6.3 CRISPR interference trials in field isolates from Xinjiang cotton regions and disease resistance data analysis The problem of yellow wilt in the cotton-growing areas of Xinjiang has a long history. Researchers have begun to use genomic and transcriptomic analyses to look for resistance clues from naturally infected strains in the field. For instance, they identified key QTLS in several different materials and screened out candidate genes significantly related to resistance (Zhang et al., 2025; Zhao et al., 2025). Some even indirectly enhance disease resistance by knocking down negative regulatory factors in cotton. These data themselves are of great value, but what is truly anticipated is the application of these findings to field CRISPR editing. Targeted intervention based on the actual pathogen population situation may enable the selection of disease-resistant varieties adapted to Xinjiang more quickly (Lei et al., 2025). 7 Prospects and Risk Assessment 7.1 Development of pathogen-targeted gene editing for green control strategies in cotton The use of gene editing techniques to deal with cotton diseases has actually been proposed for a long time. It is only in the past two years that the development of CRISPR/Cas9 has truly made it feasible. Unlike traditional transgenic methods, this approach can directly destroy the virulence factors of pathogenic bacteria without introducing exogenous DNA, or knock out the susceptibility genes (S genes) of cotton itself. The operation is relatively precise and more acceptable (Zaidi et al., 2018; Mubarik et al., 2021). For crops like cotton that are easily besieged by multiple pathogens, the effect of a single target is often insufficient. At this time, multiple editing techniques can come in handy. Once the target is expanded, the efficiency of preventing and controlling mixed infections will also be improved. This green prevention and control approach, which does not rely on pesticides and can sustain resistance, does indeed seem quite in line with the direction of sustainable agriculture (Cheng and Zhang, 2025). 7.2 Biosafety considerations in gene drive editing of microbial pathogens However, to be fair, not all seemingly "advanced" technologies can be promoted without any concerns. For instance, although gene drive technology has potential for manipulating pathogens, its risks cannot be ignored either. There are currently no complete answers to questions such as gene diffusion to non-target populations, off-target mutations, or causing some unexpected consequences in the ecosystem. Supervision must keep up. It cannot rely solely on scientists working in isolation. To enable the public to understand and accept the deployment of these technologies, it still depends on transparent communication mechanisms and rule-based evaluation systems (Pixley et al., 2019). Otherwise, even the most advanced technology may not go far due to trust issues. 7.3 Strategies for multi-pathogen interference and durable resistance in cotton To truly build lasting and effective resistance in cotton, merely editing one gene at a time is probably not enough. Pathogens are highly variable and often do not act alone. The multi-target ability of CRISPR is precisely suitable for this complex scenario. It can simultaneously modify multiple targets, such as several key S genes, or multiple pathogen-related sites, thereby reducing the possibility of resistance being "cracked" (Mubarik et al., 2021). Of course, this is not a once-and-for-all solution. It must be combined with conventional breeding methods, and dynamic monitoring of pathogenic flora is also indispensable. Only by forming linkage can this strategy be ensured to be effective in the long term in actual planting (Garcia-Ruiz et al., 2021).
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