Abstract
There are two main limitations for sprayable RNA pesticide development: delivery efficiency and synthetic cost of double-stranded RNA (dsRNA). We previously constructed a nanocarrier-based transdermal dsRNA delivery system and a novel bacteria-based hairpin RNA (hpRNA) expression system to solve these challenges. Herein, as a subsequent exploration of RNA pesticide (sprayable ds/hpRNA for pest control), we performed a greenhouse application of bacteria-expressed and nanocarrier-delivered RNA pesticide on green peach aphid. The nanoparticle SPc could combine and deliver dsRNA across the aphid cuticle and V-type proton ATPase subunits d (ATP-d) and G (ATP-G) were selected as the potential RNA interference (RNAi) targets. Our plasmid-Escherichia coli system simultaneously expressing ATP-d and ATP-G hairpin RNAs (hpRNAs) was constructed for mass production of hpRNA. The expressed hpRNA was mixed with SPc and detergent to form RNA formulation, which showed a certain insecticidal activity through the spray application in the greenhouse. Total control efficacy of our RNA pesticide could reach 61% on 3 d and maintained at 50% until the sixth day. To our knowledge, our study is the first attempt to apply the bacteria-expressed and nanocarrier-delivered RNA pesticides for pest control in the greenhouse trial, which is beneficial for promoting the development of RNA pesticides.
Highlights
RNA interference (RNAi) was first identified in the nematode Caenorhabditis elegans as a response to double-stranded RNA, resulting in the sequence-specific suppression of gene expression (Fire et al, 1998)
A gel retardation test was first performed to determine the optimal mass ratio for the combination of double-stranded RNA (dsRNA) with star polycation (SPc), and the results showed that the mass ratio of 1:1 was recommended to be used in the following study (Fig. 2A)
The fluorescent dsRNA/SPc/detergent droplet smoothly spread around the aphid integument, and high-intensity fluorescence was detected at 6 h post topical application
Summary
RNA interference (RNAi) was first identified in the nematode Caenorhabditis elegans as a response to double-stranded RNA (dsRNA), resulting in the sequence-specific suppression of gene expression (Fire et al, 1998). This technology offers new opportunities for the development of sustainable integrated pest management plans (Price and Gatehouse, 2008; Whangbo and Hunter, 2008; Bolognesi et al, 2012; Zotti and Smagghe, 2015). RNAi via sprayable or soaking approaches has been recognized as a potential strategy for pest control, and thereafter, it has been well studied in a number of insect species (Avila et al, 2018; Petek et al, 2020). Instability of dsRNA, insufficient dsRNA internalization, deficient RNAi machinery, impaired systemic spreading of the RNAi signal, and refractory target genes have been proposed as factors constraining RNAi efficiency (Wang et al, 2016; Singh et al, 2017; Christiaens et al, 2018; Cooper et al, 2019)
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