The fungus Verticillium nonalfalfae is a pathogenic species from the genus Verticillium sensu stricto, which infects hop plants and causes a disease known as Verticillium wilt. Since current control measures with fungicides have proven ineffective, we aimed to test an innovative control method called spray-induced gene silencing (SIGS) on two isolates of V. nonalfalfae, T2 and Rec. The idea is based on the mechanism of RNA interference, which regulates gene expression at the transcriptional or post-transcriptional level. By applying double-stranded RNA to the plant surface, one or more important fungal genes are silenced, thereby leading to the suppression of pathogen’s growth or even death of the pathogen. In our study, we applied a specific amount of target dsRNA onto fungal mycelia under in vitro conditions, incubated the culture plates, and at defined time points post-treatment, monitored the effect of dsRNA on mycelial growth compared to untreated controls. At each time point, we also extracted total RNA from the mycelia and used RT-qPCR to assess the expression levels of the target genes. Our results showed that the T2 isolate exhibited greater potential for gene silencing with this method, as indicated by stronger growth inhibition and a higher reduction in gene expression compared to the Rec isolate. Our findings also suggest that for effective control, continuous application of dsRNA will be necessary, ideally with encapsulation into nanoparticles to protect it from environmental degradation.
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