High-Pressure-Sprayed Double Stranded RNA Does Not Induce RNA Interference of a Reporter Gene
Open Access
- 16 December 2020
- journal article
- research article
- Published by Frontiers Media SA in Frontiers in Plant Science
- Vol. 11, 534391
- https://doi.org/10.3389/fpls.2020.534391
Abstract
In plants, RNA interference (RNAi) is an effective defense mechanism against pathogens and pests. RNAi mainly involves the micro RNA and the small interfering RNA (siRNA) pathways. The latter pathway is generally based on the processing of long double stranded RNAs (dsRNA) into siRNAs by DICER-LIKE endonucleases (DCLs). SiRNAs are loaded onto ARGONAUTE proteins to constitute the RNA-induced silencing complex (RISC). Natural dsRNAs derive from transcription of inverted repeats or of specific RNA molecules that are transcribed by RNA-directed RNA polymerase 6 (RDR6). Moreover, replication of infecting viruses/viroids results in the production of dsRNA intermediates that can serve as substrates for DCLs. The high effectiveness of RNAi both locally and systemically implicated that plants could become resistant to pathogens, including viruses, through artificial activation of RNAi by topical exogenous application of dsRNA. The most preferable procedure to exploit RNAi would be to simply spray naked dsRNAs onto mature plants that are specific for the attacking pathogens serving as a substitute for pesticides applications. However, the plant cell wall is a difficult barrier to overcome and only few reports claim that topical application of naked dsRNA triggers RNAi in plants. Using a transgenic Nicotiana benthamiana line, we found that high-pressure-sprayed naked dsRNA did not induce silencing of a green fluorescence protein (GFP) reporter gene. Small RNA sequencing (sRNA-seq) of the samples from dsRNA sprayed leaves revealed that the dsRNA was, if at all, not efficiently processed into siRNAs indicating that the dsRNA was insufficiently taken up by plant cells.Keywords
This publication has 42 references indexed in Scilit:
- Expression of Small RNA in Aphis gossypii and Its Potential Role in the Resistance Interaction with MelonPLOS ONE, 2012
- RNA Silencing and Plant Viral DiseasesMolecular Plant-Microbe Interactions®, 2012
- Spatial and functional relationships among Pol V-associated loci, Pol IV-dependent siRNAs, and cytosine methylation in the Arabidopsis epigenomeGenes & Development, 2012
- Roles of Plant Small RNAs in Biotic Stress ResponsesAnnual Review of Plant Biology, 2009
- Biochemical Evidence for Translational Repression by Arabidopsis MicroRNAsPlant Cell, 2009
- RNA interference‐inducing hairpin RNAs in plants act through the viral defence pathwayEMBO Reports, 2006
- The complex interplay between plant viruses and host RNA-silencing pathwaysCurrent Opinion in Plant Biology, 2005
- RNA silencing in plantsNature, 2004
- Cationic oligopeptide‐mediated delivery of dsRNA for post‐transcriptional gene silencing in plant cellsFEBS Letters, 2004
- Gene Expression Omnibus: NCBI gene expression and hybridization array data repositoryNucleic Acids Research, 2002