Climate change has the potential to change the distribution of pests globally and their resistance to pesticides, thereby threatening global food security in the 21st century. However, predicting where these changes occur and how they will influence current pest control efforts is a challenge. Using experimentally parameterised and field-tested models, we show that climate change over the past 50 years increased the overwintering range of a global agricultural insect pest, the diamondback moth (Plutella xylostella), by ~2.4 million km2 worldwide. Our analysis of global data sets revealed that pesticide resistance levels are linked to the species’ overwintering range: mean pesticide resistance was 158 times higher in overwintering sites compared to sites with only seasonal occurrence. By facilitating local persistence all year round, climate change can promote and expand pesticide resistance of this destructive species globally. These ecological and evolutionary changes would severely impede effectiveness of current pest control efforts and potentially cause large economic losses.

RNA interference (RNAi) has become a widely used reverse genetic tool to study gene function in eukaryotic organisms and is being developed as a technology for insect pest management. The efficiency of RNAi varies among organisms. Insects from different orders also display differential efficiency of RNAi, ranging from highly efficient (coleopterans) to very low efficient (lepidopterans). We investigated the reasons for varying RNAi efficiency between lepidopteran and coleopteran cell lines and also between the Colorado potato beetle, Leptinotarsa decemlineata and tobacco budworm, Heliothis virescens. The dsRNA either injected or fed was degraded faster in H. virescens than in L. decemlineata. Both lepidopteran and coleopteran cell lines and tissues efficiently took up the dsRNA. Interestingly, the dsRNA administered to coleopteran cell lines and tissues was taken up and processed to siRNA whereas the dsRNA was taken up by lepidopteran cell lines and tissues but no siRNA was detected in the total RNA isolated from these cell lines and tissues. The data included in this paper showed that the degradation and intracellular transport of dsRNA are the major factors responsible for reduced RNAi efficiency in lepidopteran insects.

Insect herbivores use phytochemicals as signals to induce expression of their phytochemical-detoxifying cytochrome P450 monooxygenases (P450s). The regulatory cascades that transduce phytochemical signals to enhanced expression of P450s are the focus of this review. At least seven signaling pathways, including RTK/MAPK, GPCR/CREB, GPCR/NFκB, ROS/CncC/Keap1, AhR/ARNT, cytosol NR, and nucleus-located NR, may be involved in phytochemical induction of P450s. Constitutive overexpression, overphosphorylation, and/or activation of one or more effectors in the corresponding pathway are common causes of P450 overexpression that lead to phytochemical or insecticide resistance. Future research should pay more attentions to the starting point of each pathway, the number of pathways and their cross talk for a given phytochemical, and the pathways for downregulation of P450s.Copyright © 2020 Elsevier Inc. All rights reserved.

In this editorial for the Special Issue on ‘RNAi in insect pest control’, three important applications of RNA interference (RNAi) in insects are briefly discussed and linked to the different studies published in this Special Issue. The discovery of the RNAi mechanism revolutionized entomological research, as it presented researchers with a tool to knock down genes, which is easily applicable in a wide range of insect species. Furthermore, RNAi also provides crop protection with a novel and promising pest control mode-of-action. The sequence-dependent nature allows RNAi-based control strategies to be highly species selective and the active molecule, a natural biological molecule known as double-stranded RNA (dsRNA), has a short environmental persistence. However, more research is needed to investigate different cellular and physiological barriers, such as cellular uptake and dsRNA degradation in the digestive system in insects, in order to provide efficient control methods against a wide range of insect pest species. Finally, the RNAi pathway is an important part of the innate antiviral immune defence of insects, and could even lead to applications targeting viruses in beneficial insects such as honeybees in the future.

The fall armyworm Spodoptera frugiperda (Lepidoptera: Noctuidae) is among the invasive insect pests that damages maize and sorghum, the high-priority crops in newly colonized agro-ecologies, including African contexts. Owing to the increasing infestation of the pest and the limitations of current conventional methods for its management, there is a call for discovering advanced pest management approaches. RNA interference (RNAi) is an emerging molecular tool showing flexible potential for the management of S. frugiperda. We conducted a search of the recent application of RNAi literature using Google Scholar and Mendeley to find advanced papers on S. frugiperda management using RNAi molecular tools that led to growth inhibition, developmental aberrations, reduced fecundity, and mortality, mainly by disruption of normal biological processes of the pest. Although efforts have been made to accelerate the utility of RNAi, many factors limit the efficiency of RNAi to achieve successful control over S. frugiperda. Owing to RNAi’s potential bioactivity and economic and ecological acceptability, continued research efforts should focus on improving its broad applicability, including field conditions. Screening and identification of key target genes should be a priority task to achieve effective and sustainable management of this insect via RNAi. In addition, a clear understanding of the present status of RNAi utilization in S. frugiperda management is of paramount importance to improve its efficiency. Therefore, in this review, we highlight the biology of S. frugiperda and the RNAi mechanism as a foundation for the molecular management of the pest. Then, we discuss the current knowledge of the RNAi approach in S. frugiperda management and the factors affecting the efficiency of RNAi application. Finally, the prospects for RNAi-based insect pest management are highlighted for future research to achieve effective management of S. frugiperda.

Nanoparticle-based delivery systems have emerged as powerful tools in the field of pest management, offering precise and effective means of delivering double-stranded RNA (dsRNA), a potent agent for pest control through RNA interference (RNAi). This comprehensive review aims to evaluate and compare various types of nanoparticles for their suitability in dsRNA delivery for pest management applications. The review begins by examining the unique properties and advantages of different nanoparticle materials, including clay, chitosan, liposomes, carbon, gold and silica. Each material's ability to protect dsRNA from degradation and its potential for targeted delivery to pests are assessed. Furthermore, this review delves into the surface modification strategies employed to enhance dsRNA delivery efficiency. Functionalization with oligonucleotides, lipids, polymers, proteins and peptides is discussed in detail, highlighting their role in improving stability, cellular uptake, and specificity of dsRNA delivery.This review also provides valuable guidance on choosing the most suitable nanoparticle-based system for delivering dsRNA effectively and sustainably in pest management. Moreover, it identifies existing knowledge gaps and proposes potential research directions aimed at enhancing pest control strategies through the utilization of nanoparticles and dsRNA.© 2024. The Author(s), under exclusive licence to Springer Nature B.V.

每年重大病虫害的流行和暴发都会对农业产生重大的影响，进而造成严重的经济损失，随着化学试剂的施用其抗药性也逐渐增强，因此探索新的绿色、安全、高效的害虫防治技术具有十分重要的意义。RNAi作为一种研究基因功能的工具，在开发新的害虫防治策略方面显示出巨大的潜力，但RNAi的效率受限于核酸酶的降解作用，为了改善dsRNA的有效递送，研发了纳米载导技术且被广泛应用于害虫防治。同时，基于工程菌高效合成靶向昆虫dsRNA的技术为田间推广提供了可行性及技术支撑。本文主要对RNAi技术及纳米载导RNAi技术在害虫防治中的研究和应用进行了总结和归纳。

The progress of short interfering RNA (siRNA) technologies has unlocked the development of novel alternatives for the treatment of a myriad of diseases, including viral infections, autoimmune disorders, or cancer. Nevertheless, the clinical use of these therapies faces significant challenges, mainly overcoming the charged and large nature of these molecules to effectively enter the cell. In this work, we developed a cationic polymer nanoparticle system that is able to load siRNA due to electrostatic interactions. The pH-responsiveness and membrane-disrupting ability of these carriers make them suitable intracellular delivery vehicles. In the work presented herein we synthesized, characterized, and evaluated the properties of nanoparticles based on 2-diethylaminoethyl methacrylate and tert-butyl methacrylate copolymers. A disulfide crosslinker was incorporated in the nanogels to enable the degradation of the nanoparticles in reductive environments, showing no significant changes on their physicochemical properties. The capability of the developed nanogels to be internalized, deliver siRNA, and induce gene knockdown were demonstrated using a human epithelial colorectal adenocarcinoma cell line. Overall, these findings suggest that this platform exhibits desirable characteristics as a potential siRNA-delivery platform.Copyright © 2019 Elsevier B.V. All rights reserved.

Long non-coding RNAs (lncRNAs) are transcripts without protein-coding potential that contain more than 200 nucleotides that play important roles in plant survival in response to different stresses. They interact with molecules such as DNA, RNA, and protein, and play roles in the regulation of chromatin remodeling, RNA metabolism, and protein modification activities. These lncRNAs regulate the expression of their downstream targets through epigenetic changes, at the level of transcription and post-transcription. Emerging information from computational biology and functional characterization of some of them has revealed their diverse mechanisms of action and possible roles in biological processes such as flowering time, reproductive organ development, as well as biotic and abiotic stress responses. In this review, we have mainly focused on the role of lncRNAs in biotic stress response due to the limited availability of knowledge in this domain. We have discussed the available molecular mechanisms of certain known lncRNAs against specific pathogens. Further, considering that fungal, viral, and bacterial diseases are major factors in the global food crisis, we have highlighted the importance of lncRNAs against pathogen responses and the progress in plant research to develop a better understanding of their functions and molecular mechanisms.

RNA interference (RNAi) technology is a promising approach used in pest control. The efficiency of RNAi varies considerably among different insect species, and growing evidence suggests that degradation of double-stranded RNA (dsRNA) prior to uptake is an important factor that limits RNAi efficiency in insects. Our recent work on fall webworm (Hyphantria cunea), an important invasive pest in China, showed a relatively low silencing efficiency of RNAi through dsRNA injection, which is considered the most feasible dsRNA delivery method for inducing RNAi, and the factors involved in the mechanism remain unknown. Herein, we first detected the dsRNA-degrading activity in the hemolymph and gut content of H. cunea in ex vivo assays and observed rapid degradation of dsRNA, especially in the hemolymph, which was complete within only 10 min. To determine whether dsRNA degradation could contribute to the low effectiveness of RNAi in H. cunea, four dsRNA nuclease (dsRNase) genes, HcdsRNase1, HcdsRNase2, HcdsRNase3, and HcdsRNase4, were identified by homology searching against the H. cunea transcriptome database, and their transcript levels were subsequently investigated in different tissues, developmental stages, and after dsRNA injection. Our results show that HcdsRNases are highly expressed mainly in gut tissues and hemolymph, and the expression of HcdsRNase3 and HcdsRNase4 were significantly upregulated by dsGFP induction. RNAi-of-RNAi studies, using HcCht5 as a reporter gene, demonstrated that silencing HcdsRNase3 and HcdsRNase4 significantly increases RNAi efficacy via dsHcCht5 injection, and co-silencing these two HcdsRNase genes results in a more significant improvement in efficacy. These results confirm that the RNAi efficacy in H. cunea through dsRNA injection is certainly impaired by dsRNase activity, and that blocking HcdsRNases could potentially improve RNAi, providing a reference for related studies on insects where RNAi has low efficiency.

\n Since the discovery of RNA interference (RNAi) in the nematode worm\n Caenorhabditis elegans\n in 1998 by Fire and Mello et al., strides have been made in exploiting RNAi for therapeutic applications and more recently for highly selective insect pest control. Although triggering mRNA degradation in insects through RNAi offers significant opportunities in crop protection, the application of environmental naked dsRNA is often ineffective in eliciting a RNAi response that results in pest lethality. There are many possible reasons for the failed or weak induction of RNAi, with predominant causes being the degradation of dsRNA in the formulated pesticide, in the field or in the insect once ingested, poor cuticular and oral uptake of the nucleic acid and sometimes the lack of an innate strong systemic RNAi response. Therefore, in the last 10 years significant research effort has focused on developing methods for the protection and delivery of environmental dsRNA to enable RNAi-induced insect control. This review focuses on the design and synthesis of vectors (vehicles that are capable of carrying and protecting dsRNA) that successfully enhance mRNA degradation via the RNAi machinery. The majority of solutions exploit the ability of charged polymers, both synthetic and natural, to complex with dsRNA, but alternative nanocarriers such as clay nanosheets and liposomal vesicles have also been developed. The various challenges of dsRNA delivery and the obstacles in the development of well-designed nanoparticles that act to protect the nucleic acid are highlighted. In addition, future research directions for improving the efficacy of RNA-mediated crop protection are anticipated with inspiration taken from polymeric architectures constructed for RNA-based therapeutic applications.\n

Nanoparticles have been widely applied as gene carrier for improving RNA interference (RNAi) efficiency in medical and agricultural fields. However, the mechanism and delivery process of nanoparticle-mediated RNAi is not directly visualized and elucidated. Here we synthesized a star polymer (SPc) consisted of a hydrophilic shell with positively-charged tertiary amine in the side chain, which was taken as an example to investigate the mechanism in gene delivery. The SPc could assemble with dsRNA spontaneously through electrostatic force, hydrogen bond and van der Waals force. Interestingly, the SPc could protect dsRNA from degradation by RNase A and insect hemolymph, thus remarkably increasing the stability of dsRNA. Meanwhile, the SPc could efficiently promote the cellular uptake and endosomal escape for intracellular spreading of dsRNA. Transcriptome analysis revealed that the SPc could up-regulate some key genes such as Chc, AP2S1 and Arf1 for activating clathrin-mediated endocytosis. Furthermore, the suppression of endocytosis hindered the cellular uptake of SPc-delivered dsRNA in vitro, and the subsequent RNAi effect was also disappeared in vivo. To our knowledge, our study is the first direct visualization of the detailed cellular delivery process and mechanism of nanocarrier-mediated gene delivery. Above mechanism supports the application of nanocarrier-based RNAi in gene therapy and pest management.© 2022. The Author(s).

害虫对农作物的危害严重威胁全球粮食安全，为满足日益增长的全球粮食需求，迫切需要安全有效的害虫绿色防控技术。RNAi技术（RNA interference）又叫RNA干扰技术，是一种转录后调控基因沉默的分子生物学技术，其原理是基于由19-25对核苷酸组成的小分子双链RNA与目标靶基因mRNA结合并引发mRNA降解，从而导致靶基因沉默。目前RNAi技术已被广泛应用于农作物害虫治理，在针对马铃薯靶标害虫方面，主要研究集中在防控鞘翅目、半翅目、鳞翅目害虫。2023年12月22 日，世界上首款RNAi生物农药正式批准商业化，用于防控抗药性日益严重、国际公认的马铃薯重要毁灭性检疫害虫马铃薯甲虫，是世界上首款被允许在农作物上商业使用的可喷洒RNA生物农药，对马铃薯害虫的绿色防控具有划时代的里程碑意义。基于RNAi技术的产品被用于农业害虫防控的同时，仍需考虑其抗药性、脱靶效应和对环境安全的潜在风险。本文以RNAi技术在马铃薯害虫防控应用的可行性、RNAi技术在马铃薯害虫防控中的应用以及潜在的风险等方面进行了综述，以期阐述RNAi技术在马铃薯害虫防控中的应用现状与前景，为RNAi技术纳入防控马铃薯害虫综合治理体系提供理论参考。

RNA干扰（RNAi）是生物体内源基因发生转录后特异性降解的一种生理现象,广泛存在于生物体内。RNAi主要由小干扰RNA诱发阻碍目的基因的翻译或转录,造成目标信使RNA沉默。RNAi具有高效、特异性强等优点,被广泛应用于昆虫基因功能研究,并显示出了开发新型病虫害管理策略的巨大潜力。主要阐述了RNAi的沉默机制,双链RNA转入昆虫体内的几种方式,以及RNAi技术在不同目昆虫中研究的最新进展。最后,对RNAi技术存在的不足之处进行了简单总结,还对RNAi技术在害虫防治中的应用进行了展望,以期为该技术广泛应用于农业害虫防治提供理论支持。

RNA interference is a phenomenon in which the introduction of double-stranded RNA (dsRNA) into cells triggers the degradation of the complementary messenger RNA in a sequence-specific manner. Suppressing expression of vital genes could lead to insect death, therefore this technology has been considered as a potential strategy for insect pest control. There are three main routes of dsRNA administration into insects: (i) injections to the hemolymph, (ii) topical, and (iii) feeding. In this review, we focus on dsRNA administration through feeding. We summarize novel strategies that have been developed to improve the efficacy of this method, such as the use of nano-based formulations, engineered microorganisms, and transgenic plants. We also expose the hurdles that have to be overcome in order to use this technique as a reliable pest management method. © 2019 Society of Chemical Industry.© 2019 Society of Chemical Industry.

RNA interference (RNAi) has proven to be a very promising prospect for insect pest control. However, low RNAi efficacy limits further development of this biotechnology for use on lepidopteran insects, including the rice striped stem borer (SSB) (Chilo suppressalis), one of the major destructive rice pests. In this work, the application of various nanoparticles (NPs) by which double-stranded RNA (dsRNA) could be encapsulated was evaluated as an alternative delivery strategy to potentially increase the bioactivity of dsRNA. Three NPs, chitosan, carbon quantum dot (CQD), and lipofectamine2000, complexed with dsRNA (to target the glyceraldehyde-3-phosphate dehydrogenase gene (G3PDH)) were tested to examine their use in controlling SSB. Relative mRNA expressions were quantified using qPCR to evaluate knockdown efficiency of NP-dsRNA treated larvae, and the correlated dsRNA-mediated SSB larval mortality was tested. Thereafter, the content dynamics of hemolymph dsRNA after ingesting different NP-dsRNA were monitored in vivo; the hemolymph dsRNA content was in ratios of 5.67, 9.43, and 1 with chitosan, CQD, and lipofectamine2000 induced samples, respectively. The results demonstrated that all three tested NPs led to efficient feeding delivery by improving both dsRNA stability and cellular uptake equally. Furthermore, there was a strong correlation (r= 0.9854) between the hemolymph dsRNA contents and the average RNAi depletions in the non-gut tissues of SSB. Overall, our results strongly suggest that due to its strong endosomal escaping ability, CQD was the most efficient carrier for inducing systemic RNAi, and thereby causing effective gene silencing and mortality in SSB.Copyright © 2019 Elsevier Inc. All rights reserved.

Polyamidoamine (PAMAM) dendrimers are emerging as intriguing nanovectors for nucleic acid delivery because of their unique well-defined architecture and high binding capacity, which have been broadly applied in DNA- and RNA-based therapeutics. The low-cost and high-efficiency of PAMAM dendrimers relative to traditional liposomal transfection reagents also promote their application in gene function analysis. In this study, we first investigated the potential use of a PAMAM system in the silkworm model insect. We determined the binding property of G5-PAMAM using dsRNA and DNA in vitro, and substantially achieved the delivery of dsRNA and DNA from culture medium to both silkworm BmN and BmE cells, thus leading to efficient knockdown and expression of target genes. Under treatments with different concentrations of G5-PAMAM, we evaluated its cellular cytotoxicity on silkworm cells, and the results show that G5-PAMAM had no obvious toxicity to cells. The presence of serum in the culture medium did not affect the delivery performance of DNA and dsRNA by G5-PAMAM, revealing its convenient use for various purposes. In conclusion, our data demonstrate that the PAMAM system provides a promising strategy for delivering dsRNA and DNA in cultured silkworm cells and promote its further application in individuals.

Chickpea pod borer,, displays resistance to chemical insecticides and transgenics. The potential nontransformative RNAi approach of specific gene silencing by mRNA breakdown through exogenous double-stranded (dsRNA) delivery to faces problems of degradation by nucleases and insect gut pH. We demonstrate that chitosan nanoparticles (CNPs) effectively mediate specific dsRNA delivery against () and ( target genes. Ionotropically synthesized cationic CNPs (100 nm size, +32 mV charge) loaded dsRNA efficiently and protected it effectively from degradation by nucleases and insect gut pH. Tagging CNPs with Calcofluor fluorescence illustrated its efficient uptake in columnar insect gut cells. The potential of CNPs-mediated dsRNA delivery was elucidated with effective silencing of green fluorescent protein transformed Sf9 cells. Furthermore, CNPs-dsRNA complexes were stable for 5 d on leaf surfaces, and their ingestion with leaf effectively silenced and genes to suppress related enzyme activities and caused 100% insect mortality. Further, in planta bioassay with CNPs-dsRNA spray confirmed the RNAi induced insect mortality. Moreover, CNPs-dsRNA fed nontarget insects and were unaffected, and no toxicity was observed for CNPs in cell line studies. Remarkably, only two low dose (0.028 g/ha) topical CNPs-ache-dsRNA sprays on chickpea displayed reduced pod damage with high yields on par with chemical control in the field, which was followed by CNPs-jhamt-dsRNA nanoformulation. These studies can pave the way for the development of topical application of CNPs-dsRNA spray as a safe, specific, innovative insecticide for sustainable crop protection.

RNAi technology, known as a revolutionary technology in the history of pesticides, has been identified as a very promising novel approach for crop protection, which is of great significance for achieving the sustainable agricultural development of the United Nations Food and Agriculture Organization. Although many studies have shown that RNA biopesticides have strong application prospects, its stability seriously restricts the commercial use. As the core component of RNAi, double-stranded RNA (dsRNA) is unstable in its natural form. Therefore, how to ensure the stability of dsRNA is one of the most significant challenges in realizing the commercial use of RNA biopesticides. Nanomaterials such as cationic polymers and lipofectamine can improve the stability of dsRNA in the environment, which has been proved. This paper reviews the recent research progress of nanomaterials that can be used to improve the environmental stability of dsRNA, and discusses the advantages and limitations of different nanomaterials combined with dsRNA, which provides reference for the selection of dsRNA nanoformulations.