Advances in the Development of Microbial Double-Stranded RNA Production Systems for Application of RNA Interference in Agricultural Pest Control

Guan, Ruobing; Chu, Dongdong; Han, Xinyi; Xing, Miao; Li, Haichao

doi:10.3389/fbioe.2021.753790

Cited by 30 publications

(19 citation statements)

References 69 publications

(80 reference statements)

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“…Using RNAi for pest management requires the introduction of dsRNA in vivo to knockdown genetic targets of interest, thereby activating the siRNA pathway. By silencing specific genes, the roles of multiple functional genes in the sensing system have been elucidated ( Huvenne and Smagghe, 2010 ; Zhao et al, 2011 ; Guan et al, 2021 ; Zhan et al, 2021 ). RNAi can also be used for pest control, as an effective alternative to chemical pesticides.…”

Section: Introductionmentioning

confidence: 99%

Silencing the gustatory receptor BtGR11 affects the sensing of sucrose in the whitefly Bemisia tabaci

Tian

et al. 2022

Front. Bioeng. Biotechnol.

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RNA interference (RNAi) is powerful biotechnology for studying the in vivo functions of key genes. Based on this property, RNAi can also be used for pest control as an effective alternative to chemical pesticides. The management of phloem-sucking pests is a tricky issue in current agricultural and forestry pest control. RNAi can silence key chemoreceptor genes of phloem-sucking pests; thereby regulating the behavior of these pests can be manipulated. So, it is considered to be a promising new type of ecological pest management strategy. In this study, we identified a candidate taste receptor gene, BtGR11, that controls the taste sensitivity to sucrose in the whitefly Bemisia tabaci, which is a serious invasive phloem-sucking pest worldwide. Functional analyses using the Xenopus oocyte expression system and the two-electrode voltage-clamp system revealed that the oocytes expressing BtGR11 responded to sucrose. Furthermore, we found that silencing BtGR11 by RNAi inhibited the function of sensing sucrose in the whitefly. This study reports a key chemoreceptor gene that can be used for the understanding of the gustatory sensing mechanisms of whitefly to deterrent.

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Section: Introductionmentioning

confidence: 99%

Silencing the gustatory receptor BtGR11 affects the sensing of sucrose in the whitefly Bemisia tabaci

Tian

et al. 2022

Front. Bioeng. Biotechnol.

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“…However, the engineered microbe may induce an undesirable host immune response. There is also a signi cant risk of environmental proliferation of the microbe (Guan et al, 2021) and the potential for interspecies transfer of plasmid-based expression elements that may lead to sustained dsRNA expression (Mendelsohn et al, 2020) that may impact non-target organisms. From this perspective, sprayable formulated ''naked'' bacterial-derived dsRNA is more desirable associated puri cation cost is a signi cant challenge.…”

Section: Discussionmentioning

confidence: 99%

A novel sustainable platform for scaled manufacturing of double-stranded RNA biopesticides

Nwokeoji

Chou³

et al. 2022

Preprint

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RNA interference (RNAi) represents one of the most conserved pathways evolved by eukaryotic cells for regulating gene expression. RNAi utilises non-translatable double-stranded RNA (dsRNA) molecules to sequester or degrade mRNA molecules gene. In RNAi, specifically-designed exogenous dsRNA delivered to the cell can silence a target gene, a phenomenon that has been exploited in many functional studies and explored in biopesticide applications. The search for safe and sustainable crop pest management options drives the need to offset the effect of inorganic pesticides on biodiversity. The prospect of replacing inorganic pesticides with dsRNA crop spray is gaining popularity, enhanced by its high-target specificity and low-environmental impact. However, for dsRNA to reach the pesticide market, it must be produced cost-effectively and sustainably. In this paper, we develop a high-yield expression media that generates up to 15-fold dsRNA yield compared to existing expression media utilising 1mM IPTG. We also optimise a low-cost purification method that generates high-quality and purified dsRNA. The developed method circumvents the need for hazardous chemical reagents often found in commercial kits or commercial nucleases to eliminate contaminating DNA or single-stranded RNA (ssRNA) species. We also demonstrate that the production platform is scalable, generating 6.29 mg dsRNA from 259 mg wet E. coli cell pellet. The results also provide structural insights into the heterogeneous dsRNA species within the microbial-derived dsRNA pool. Finally, we also show that the purified 'naked' dsRNA, without prior formulation, can induce insect toxicity under field conditions. This study provides a novel, complete, low-cost process dsRNA platform with potential for application in industrial dsRNA production.

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“…This novel, foliar-applied biopesticide represents a new mode-of-action class of pesticides. Bayer's (formerly Monsanto) BioDirect ™ also represents a SIGS-based bioproduct, targeting Varroa destructor mites for the protection of honeybees (Guan et al, 2021;De Schutter et al, 2022).…”

Section: Direct Deliverymentioning

confidence: 99%

Plant biomacromolecule delivery methods in the 21st century

et al. 2022

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The 21st century witnessed a boom in plant genomics and gene characterization studies through RNA interference and site-directed mutagenesis. Specifically, the last 15 years marked a rapid increase in discovering and implementing different genome editing techniques. Methods to deliver gene editing reagents have also attempted to keep pace with the discovery and implementation of gene editing tools in plants. As a result, various transient/stable, quick/lengthy, expensive (requiring specialized equipment)/inexpensive, and versatile/specific (species, developmental stage, or tissue) methods were developed. A brief account of these methods with emphasis on recent developments is provided in this review article. Additionally, the strengths and limitations of each method are listed to allow the reader to select the most appropriate method for their specific studies. Finally, a perspective for future developments and needs in this research area is presented.

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Advances in the Development of Microbial Double-Stranded RNA Production Systems for Application of RNA Interference in Agricultural Pest Control

Cited by 30 publications

References 69 publications

Silencing the gustatory receptor BtGR11 affects the sensing of sucrose in the whitefly Bemisia tabaci

Silencing the gustatory receptor BtGR11 affects the sensing of sucrose in the whitefly Bemisia tabaci

A novel sustainable platform for scaled manufacturing of double-stranded RNA biopesticides

Plant biomacromolecule delivery methods in the 21st century

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