Selection of BcTRE1 as an effective RNAi target for dsRNA-Based control of Botrytis cinerea

RNA interference (RNAi) is emerging as a promising approach for plant disease management. In this study, we screened five virulence-related genes of Botrytis cinerea to identify the most effective target for controlling B. cinerea through RNAi pathway. The double-stranded RNA (dsRNA) was synthesized in Escherichia coli and evaluated for their efficacy in suppressing spore germination, mycelial growth, and virulence in infected plants. Our results demonstrated that BcTRE1-targeting dsRNA (BcTRE1-dsRNA) exhibited the highest inhibitory activity, significantly reducing fungal growth and lesion formation compared to the other tested dsRNAs. Gene expression analysis confirmed that BcTRE1-dsRNA effectively silenced BcTRE1 expression within 7 days post-treatment, aligning with the transient protection window observed for naked dsRNA applications. To extend this protective period, we incorporated layered double hydroxide (LDH) nanocarriers for dsRNA delivery, which successfully prolonged the inhibitory effect, reducing lesion formation even at 11 days post-treatment. These findings identify BcTRE1 as a key RNAi target and highlight the potential of dsRNA-LDH formulations for sustainable fungal disease management in crops.