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      Improving RNAi efficiency for pest control in crop species

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          Abstract

          The application of RNAi promotes the development of novel approaches toward plant protection in a sustainable way. Genetically modified crops expressing dsRNA have been developed as commercial products with great potential in insect pest management. Alternatively, some nontransformative approaches, including foliar spray, irrigation and trunk injection, are favorable in actual utilization. In this review, we summarize the recent progress and successful cases of RNAi-based pest management strategy, explore essential implications and possibilities to improve RNAi efficiency by delivery of dsRNA through transformative and nontransformative approaches, and highlight the remaining challenges and important issues related to the application of this technology.

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          Control of coleopteran insect pests through RNA interference.

          James Baum, Thierry Bogaert, William Jefferson Clinton … (2007)
          Commercial biotechnology solutions for controlling lepidopteran and coleopteran insect pests on crops depend on the expression of Bacillus thuringiensis insecticidal proteins, most of which permeabilize the membranes of gut epithelial cells of susceptible insects. However, insect control strategies involving a different mode of action would be valuable for managing the emergence of insect resistance. Toward this end, we demonstrate that ingestion of double-stranded (ds)RNAs supplied in an artificial diet triggers RNA interference in several coleopteran species, most notably the western corn rootworm (WCR) Diabrotica virgifera virgifera LeConte. This may result in larval stunting and mortality. Transgenic corn plants engineered to express WCR dsRNAs show a significant reduction in WCR feeding damage in a growth chamber assay, suggesting that the RNAi pathway can be exploited to control insect pests via in planta expression of a dsRNA.
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            Silencing a cotton bollworm P450 monooxygenase gene by plant-mediated RNAi impairs larval tolerance of gossypol.

            Ying-Bo Mao, Wen-Juan Cai, Jia-Wei Wang … (2007)
            We identify a cytochrome P450 gene (CYP6AE14) from cotton bollworm (Helicoverpa armigera), which permits this herbivore to tolerate otherwise inhibitory concentrations of the cotton metabolite, gossypol. CYP6AE14 is highly expressed in the midgut and its expression correlates with larval growth when gossypol is included in the diet. When larvae are fed plant material expressing double-stranded RNA (dsRNA) specific to CYP6AE14, levels of this transcript in the midgut decrease and larval growth is retarded. Both effects are more dramatic in the presence of gossypol. As a glutathione-S-transferase gene (GST1) is silenced in GST1 dsRNA-expressing plants, feeding insects plant material expressing dsRNA may be a general strategy to trigger RNA interference and could find applications in entomological research and field control of insect pests.
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              Passenger-strand cleavage facilitates assembly of siRNA into Ago2-containing RNAi enzyme complexes.

              Christian Matranga, Yukihide Tomari, Chanseok Shin … (2005)
              In the Drosophila and mammalian RNA interference pathways, siRNAs direct the protein Argonaute2 (Ago2) to cleave corresponding mRNA targets, silencing their expression. Ago2 is the catalytic component of the RNAi enzyme complex, RISC. For each siRNA duplex, only one strand, the guide, is assembled into the active RISC; the other strand, the passenger, is destroyed. An ATP-dependent helicase has been proposed first to separate the two siRNA strands, then the resulting single-stranded guide is thought to bind Ago2. Here, we show that Ago2 instead directly receives the double-stranded siRNA from the RISC assembly machinery. Ago2 then cleaves the siRNA passenger strand, thereby liberating the single-stranded guide. For siRNAs, virtually all RISC is assembled through this cleavage-assisted mechanism. In contrast, passenger-strand cleavage is not important for the incorporation of miRNAs that derive from mismatched duplexes.
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                Author and article information

                Journal
                Journal ID (nlm-ta): Biotechniques
                Journal ID (iso-abbrev): BioTechniques
                Journal ID (publisher-id): BTN
                Title: Biotechniques
                Publisher: Future Science Ltd (London, UK )
                ISSN (Print): 0736-6205
                ISSN (Electronic): 1940-9818
                Publication date (Electronic, pub): 23 March 2020
                Publication date (Print, pub): May 2020
                Publication date PMC-release: 23 March 2020
                Volume: 68
                Issue: 5
                Pages: 283-290
                Affiliations
                [1 ]Department of Entomology, MOA Key Lab of Pest Monitoring & Green Management, College of Plant Protection, China Agricultural University, Beijing, People’s Republic China
                [2 ]National Agricultural Technology Extension & Service Center, Beijing, People’s Republic China
                Author notes
                [* ]Author for correspondence: shenjie@ 123456cau.edu.cn
                Article
                DOI: 10.2144/btn-2019-0171
                PMC ID: 7252490
                PubMed ID: 32202134
                SO-VID: e9375199-8ebd-4a5b-af96-1ef2c03d0941
                Copyright © © 2020 Shuo Yan
                License:

                This work is licensed under the Attribution-NonCommercial-NoDerivatives 4.0 Unported License

                History
                Date received : 28 December 2019
                Date accepted : 19 February 2020
                Date: 23 March 2020
                Page count
                Pages: 8
                Categories
                Subject: Review

                Keywords: dsrna delivery,foliar spray,nanoparticle,nontransformative rnai product,rnai-based pest management,rnai efficiency,transformative rnai product,transplastomic crop
                Data availability:
                Keywords: dsrna delivery, foliar spray, nanoparticle, nontransformative rnai product, rnai-based pest management, rnai efficiency, transformative rnai product, transplastomic crop

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