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Artificial nanovesicles for dsRNA delivery in spray-induced gene silencing for crop protection |
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| Authors: | Lulu Qiao Jonatan Niño-Sánchez Rachael Hamby Luca Capriotti Angela Chen Bruno Mezzetti Hailing Jin |
| Affiliation: | 1. Department of Microbiology & Plant Pathology, Center for Plant Cell Biology, Institute for Integrative Genome Biology, University of California, Los Angeles, CA, USA
State Key Laboratory of Plant Physiology and Biochemistry, College of Life Sciences, Zhejiang University, Hangzhou, China These authors contributed equally to this work.;2. Department of Microbiology & Plant Pathology, Center for Plant Cell Biology, Institute for Integrative Genome Biology, University of California, Los Angeles, CA, USA Department of Plant Production and Forest Resources, University of Valladolid, Palencia, Spain Sustainable Forest Management Research Institute (iuFOR), University of Valladolid, Palencia, Spain These authors contributed equally to this work.;3. Department of Microbiology & Plant Pathology, Center for Plant Cell Biology, Institute for Integrative Genome Biology, University of California, Los Angeles, CA, USA These authors contributed equally to this work.;4. Department of Agricultural, Food and Environmental Sciences, Marche Polytechnic University, Ancona, Italy;5. Department of Microbiology & Plant Pathology, Center for Plant Cell Biology, Institute for Integrative Genome Biology, University of California, Los Angeles, CA, USA |
| Abstract: | 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. |
| Keywords: | spray-induced gene silencing small RNA RNA interference double-stranded RNA uptake efficiency nanoparticles |
