Double-Stranded RNA-Mediated Interference with Plant Virus Infection

Abstract

Double-stranded RNA (dsRNA) has been shown to play a key role as an inducer of different interference phenomena occurring in both the plant and animal kingdoms. Here, we show that dsRNA derived from viral sequences can interfere with virus infection in a sequence-specific manner by directly delivering dsRNA to leaf cells either by mechanical inoculation or via an Agrobacterium-mediated transient-expression assay. We have successfully interfered with the infection of plants by three viruses belonging to the tobamovirus, potyvirus, and alfamovirus groups, demonstrating the reliability of the approach. We suggest that the effect mediated by dsRNA in plant virus infection resembles the analogous phenomenon of RNA interference observed in animals. The interference observed is sequence specific, is dose dependent, and is triggered by dsRNA but not singlestranded RNA. Our results support the view that a dsRNA intermediate in virus replication acts as efficient initiator of posttranscriptional gene silencing (PTGS) in natural virus infections, triggering the initiation step of PTGS that targets viral RNA for degradation.

Short interfering RNA-mediated interference of gene expression and viral DNA accumulation in cultured plant cells

“…Recently it was shown that siRNAs of 21 nt in size, an intermediate of the RNA-interference pathway, are effective in suppression of gene expression in animal systems including mammals. Gene silencing induced by long dsRNA constructs has been demonstrated in transgenic plants showing resistance to virus infection (9): transient interference with endogenous gene expression (28), with virus infection in plants (29), and with reporter gene expression in plant cells (30,31). The siRNA technology offers animal virologists a new way to control viruses because the use of long dsRNA triggers the IFN response, which leads to apoptosis in most mammalian cells (32).…”

Phyllotreta striolata (Coleoptera: Chrysomelidae): Arginine kinase cloning and RNAi-based pest control

“…Previous studies indicate that dsRNA molecules are highly stable in vitro and can persist on the leaves for several days (Tenllado & Diaz-Ruiz, 2001). Thus RNAi technology could be deployed in the field.…”

“…RNAi has become a potent tool in many biological research fields ranging from identifying gene function to inhibiting viral infection (Tenllado & Diaz-Ruiz, 2001;Tenllado et al, 2003a). Insect genes can be downregulated by microinjection of dsRNA (Bettencourt et al, 2002;Tomoyasu & Denell, 2004;Ghanima et al, 2007) or uptake of exogenous dsRNA from an artificial diet (Turner et al, 2006).…”

Single Nucleotide Modification of VP2 Gene of Highly Virulent Infectious Bursal Disease Virus for Effective Expression Strategy in Tobacco

“…In productive viral infections, most viruses can counteract the RNA silencing defense, in part by expressing proteins that interfere with the RNA-silencing machinery [43,44]. Silencing of the transgene would target viral RNAs containing enough sequence similarity to the transgene for degradation, therefore inhibiting virus infection [45,46]. The expressed VP2 protein was further analyzed by Western blot analyses for the confirmation of protein expression.…”