Abstract
RNA interference (RNAi)-based pesticides are pest control agents that use RNAi mechanisms as the basis of their action. They are regarded as environmentally friendly and are a promising alternative to conventional chemical pesticides. The effective substance in RNAi-based pesticides is double-stranded RNA (dsRNA) designed to match the nucleotide sequence of a target essential gene of the pest of concern. When taken up by the pest, this exerts an RNAi effect and inhibits some vital biochemical/biological process in the pest. dsRNA products are also expected to be applied for the control of viral diseases in aquaculture by RNAi, especially in shrimp farming. A critical issue in the practical application of RNAi agents is that production of the dsRNA must be low-cost. Here, we review recent methods for microbial production of dsRNAs using representative microorganisms (Escherichia coli, Pseudomonas syringae, Corynebacterium glutamicum, Chlamydomonas reinhardtii, and others) as host strains. The characteristics of each dsRNA production system are discussed.
Highlights
As the global population and living standards grow, production of abundant, highquality, safe food is increasingly important
Aalto et al [55] employed the egfp gene as a target double-stranded RNA (dsRNA) example for genetic knockdown experiments, allowing the egfp gene flanked by 50 -packaging and 30 -replication signals to be expressed by T7 RNA polymerase on plasmid pPS9, and introduced these into the host microbial cells. egfp-ssRNAs were converted to dsRNAs inside empty polymerase complex (PC), and the production of capsids containing egfp-dsRNAs (1.6 mg dsRNA/g wet cells) was achieved in P. syringae
The antiviral protective effect of using killed bacteria containing the target dsRNA was comparable to that of using dsRNA extracted from the producing strain. These findings suggest that the sterilization of dsRNA-producing microbes by alcohol treatment and the resultant encapsulation of dsRNA may be effective for the formulation of RNA interference (RNAi)-based pesticides in a variety of dsRNA-producing microbes
Summary
As the global population and living standards grow, production of abundant, highquality, safe food is increasingly important. In aquaculture, especially shrimp farming, concerns have been raised about the ecological impact of the use of large amounts of chemical antibiotics and synthetic antiviral agents that can induce the emergence of drug-resistant pathogens, and the development of more natural and environmentally friendly disease-control agents is coveted. A second advantage is that the RNA, the effective factor of the insecticide, degrades reasonably quickly in the environment because of its inherent chemical instability and the ubiquitous presence of nucleases in the bioecological community [14,15] This feature of RNAi-based pesticides is fundamentally consistent with protection of ecosystems. Recent research cases of dsRNA-production using various yeasts, lactic acid bacteria, and symbiotic microbes have been reported for applications in pest control
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