Artificial nanovesicles for dsRNA delivery in spray‐induced gene silencing for crop protection

• Published in: Plant biotechnology journal Vol. 21; no. 4; pp. 854 - 865
• Main Authors: Qiao, Lulu, Niño‐Sánchez, Jonatan, Hamby, Rachael, Capriotti, Luca, Chen, Angela, Mezzetti, Bruno, Jin, Hailing
• Format: Journal Article
• Language: English
• Published: England John Wiley & Sons, Inc 01.04.2023; John Wiley and Sons Inc
• Subjects: Agricultural production; Biosynthesis; Clean technology; Crop diseases; Crop Protection - methods; Disease control; Double-stranded RNA; Encapsulation; Environmental conditions; Food security; Fungicides; Gene Silencing; Genes; Genomic instability; Leaves; Lipids; Nanoparticles; Nuclease; Pathogens; Pesticides; Plant diseases; Plant protection; Proteins; Ribonucleic acid; RNA; RNA Interference; RNA, Double-Stranded - genetics; RNA-mediated interference; small RNA; spray‐induced gene silencing; Tomatoes; Topical application; uptake efficiency; Virulence; RNA interference; nanoparticles; uptake efficiency; double-stranded RNA; small RNA; spray-induced gene silencing
• ISSN: 1467-7644; 1467-7652
• DOI: 10.1111/pbi.14001

Summary Spray‐induced gene silencing (SIGS) is an innovative and eco‐friendly technology where topical application of pathogen gene‐targeting RNAs to plant material can enable disease control. SIGS applications remain limited because of the instability of RNA, which can be rapidly degraded when exposed to various environmental conditions. Inspired by the natural mechanism of cross‐kingdom RNAi through extracellular vesicle trafficking, we describe herein the use of artificial nanovesicles (AVs) for RNA encapsulation and control against the fungal pathogen, Botrytis cinerea. AVs were synthesized using three different cationic lipid formulations, DOTAP + PEG, DOTAP and DODMA, and examined for their ability to protect and deliver double stranded RNA (dsRNA). All three formulations enabled dsRNA delivery and uptake by B. cinerea. Further, encapsulating dsRNA in AVs provided strong protection from nuclease degradation and from removal by leaf washing. This improved stability led to prolonged RNAi‐mediated protection against B. cinerea both on pre‐ and post‐harvest plant material using AVs. Specifically, the AVs extended the protection duration conferred by dsRNA to 10 days on tomato and grape fruits and to 21 days on grape leaves. The results of this work demonstrate how AVs can be used as a new nanocarrier to overcome RNA instability in SIGS for crop protection.