Abstract
At the present time, the control of plant viral diseases is primarily achieved through the use of traditional pesticides. However, the tendency of RNA viruses to mutate and develop resistance to pesticides represents a significant challenge. The development of nanoscale strategies based on RNA interference (RNAi) represents a rapidly evolving approach to sustainable plant disease control. In this study, an siRNA was screened that effectively targets the 70-kDa heat shock protein (HSP70), which is closely associated with plant virus infection and replication. Additionally, a novel nanosystem for hierarchical delivery of siRNA was developed. This consisted of the cellular shuttle peptide (CPP)-modified tetrahedral DNA nanoparticles (TDN) as the core and polydopamine-hybridized mesoporous silica nanoparticles (PDA-MSN) as the shell. This was named TDN-siR099@CPP@PDA-MSNs. The results demonstrated that upon entering the leaf tissue through the stomata, TDN-siR099@CPP@PDA-MSNs encountered a weak acidic environment, prompting the PDA-MSN to release TDN-siR099@CPP. This resulted in the intercellular shuttling of TDN-siR099@CPP. The nanoscale dimensions of TDN-siR099@CPP facilitate facile displacement, while the intrinsic properties of CPP enhance the efficacy of long-distance siR099 intercellular shuttling. In this nanosystem, we also demonstrated that PDA-MSN retarded the susceptible degradation of siRNA in vitro and responded to the acidic release of the drug, which improve the utilization efficiency of siRNA in terms of both improving the delivery efficiency and delaying the degradation. It effectively down-regulated 56.2% of HSP70 protein and inhibited the infection of tobacco mosaic virus (TMV) and potato Y virus (PVY) by 70.2% and 41.0%, respectively. By skillfully constructing a framework for siRNA inhibition of target genes, this technology broadens the application of RNA silencing in plant disease management.
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