Physiological Conditions and dsRNA Application Approaches for Exogenously induced RNA Interference in Arabidopsis thaliana

• Published in: Plants (Basel) Vol. 10; no. 2; p. 264
• Main Authors: Kiselev, Konstantin V, Suprun, Andrey R, Aleynova, Olga A, Ogneva, Zlata V, Dubrovina, Alexandra S
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 30.01.2021; MDPI
• Subjects: Arabidopsis thaliana; Diurnal variations; Double-stranded RNA; exogenously induced RNA interference (exo-RNAi); external dsRNA; foliar application; Foliar applications; Gene expression; Gene silencing; Genes; Infiltration; Inoculation; Interference; Nanoparticles; Neomycin; Neomycin phosphotransferase; Pathogens; Peptides; Phosphotransferase; Phosphotransferase II; Physiology; plant gene silencing; Plant resistance; Ribonucleic acid; RNA; RNA-mediated interference; siRNA; Soil conditions; Soil moisture; Spraying; Time of use; transgene; Transgenes; Transgenic plants; Arabidopsis thaliana; exogenously induced RNA interference (exo-RNAi); transgene; external dsRNA; plant gene silencing; foliar application
• ISSN: 2223-7747; 2223-7747
• DOI: 10.3390/plants10020264

Recent studies have revealed that foliar application of double-stranded RNAs (dsRNAs) or small-interfering RNAs (siRNAs) encoding specific genes of plant pathogens triggered RNA interference (RNAi)-mediated silencing of the gene targets. However, a limited number of reports documented silencing of plant endogenes or transgenes after direct foliar RNA application. This study analyzed the importance of physiological conditions (plant age, time of day, soil moisture, high salinity, heat, and cold stresses) and different dsRNA application means (brush spreading, spraying, infiltration, inoculation, needle injection, and pipetting) for suppression of neomycin phosphotransferase II ( ) transgene in , as transgenes are more prone to silencing. We observed a higher suppression when dsRNA was applied at late day period, being most efficient at night, which revealed a diurnal variation in dsRNA treatment efficacy. Exogenous -dsRNA considerably reduced expression in 4-week-old plants and only limited it in 2- and 6-week-old plants. In addition, a more discernible downregulation was detected under low soil moisture conditions. Treatment of adaxial and abaxial leaf surfaces by brushes, spraying, and pipetting showed a higher suppression, while infiltration and inoculation were less efficient. Thus, appropriate plant age, late time of day, low soil moisture, and optimal dsRNA application modes are important for exogenously induced gene silencing.