Abstract
Hairpin RNAs have been used to confer resistance to viruses in plants through RNA silencing. However, it has not been demonstrated that RNA silencing was effective against inoculation by aphids of non-persistently transmitted viruses, the major route of plant virus spread in nature. As a proof-of-principle strategy, we made use of Agrobacterium tumefaciens to transiently express a hairpin RNA homologous to Potato virus Y (PVY) in plant tissues. A complete and specific interference with aphid transmission of PVY was achieved by inducers of RNA silencing, as demonstrated by specific siRNAs accumulation in agroinfiltrated tissues. To our knowledge, this is the first report of successful interference with non-persistent transmission of a plant virus using RNA interference.
Highlights
One of the most efficient mechanisms by which plants protect themselves from viruses is the specific RNAdependent silencing pathway termed post-transcriptional gene silencing (PTGS)
The RNA silencing machinery recognizes several features of viral infections involving the formation of double-stranded RNA and initiates a response that degrades viral RNA and eventually enables the plant to recover from virus infection [1]
Several studies have demonstrated that inverted repeat constructs encoding self-complementary RNAs can effectively induce RNA silencing and lead to high resistance frequencies in transgenic plants [2,3]
Summary
One of the most efficient mechanisms by which plants protect themselves from viruses is the specific RNAdependent silencing pathway termed post-transcriptional gene silencing (PTGS). We and others have previously shown that exogenously supplied dsRNA, or vectors expressing it, derived from viral sequences can interfere with virus infection in non-transgenic plants [57]. The invading virus containing sequences homologous to the dsRNA is recognized and degraded by the plant's defence mechanism This non-transgenic, RNAi-based approach could form the basis for the development of a new biotechnological tool aimed at protecting crops against virus diseases [9]. We previously showed that transient expression of a hairpin RNA could block multiplication and spread of a tobamovirus delivered by mechanical inoculation in non-transgenic plants [12]. We investigated the silencing potential of inverted repeat sequences designed to generate a hairpin RNA homologous to Potato virus Y (PVY) for its ability to interfere with the non-persistent transmission of this virus by aphids. Dot blot analysis of total soluble proteins extracted from systemic leaves at 14 dpi
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