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1.
BACKGROUND: RNA interference (RNAi) is commonly used in insect functional genomics studies and usually involves direct injection of double‐stranded RNA (dsRNA). Only a few studies have involved exposure to dsRNAs through feeding. For western corn rootworm (Diabrotica virgifera virgifera) larvae, ingestion of dsRNA designed from the housekeeping gene, vacuolar ATPase (vATPase) triggers RNAi causing growth inhibition and mortality; however, the effect of dsRNA feeding on adults has not been examined. In this research, WCR adults were fed with vATPase‐dsRNA‐treated artificial diet containing a cucurbitacin bait, which is a proven feeding stimulant for chrysomelid beetles of the subtribe Diabroticina to which rootworms belong. RESULTS: Real‐time PCR confirmed suppression of vATPase expression and western blot analysis indicated reduced signal of a protein that cross‐reacted with a vATPase polyclonal antiserum in WCR adults exposed to artificial diet treated with dsRNA and cucurbitacin bait. Continuous feeding on cucurbitacin and dsRNA‐treated artificial diet resulted in more than 95% adult mortality within 2 weeks while mortality in control treatments never exceeded 20%. CONCLUSIONS: This research clearly demonstrates the effect of RNAi on WCR adults that have been exposed to dsRNA by feeding and establishes a tool to screen dsRNAs of potential target genes in adults. This technique may serve as an alternative to target screening of larvae which are difficult to maintain on artificial diets. Copyright © 2011 Society of Chemical Industry 相似文献
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Xiu‐Dao Yu Zong‐Cai Liu Si‐Liang Huang Zhi‐Qin Chen Yong‐Wei Sun Peng‐Fei Duan You‐Zhi Ma Lan‐Qin Xia 《Pest management science》2016,72(6):1090-1098
Aphids (Aphididae) are major agricultural pests that cause significant yield losses of crop plants each year by inflicting damage both through the direct effects of feeding and by vectoring harmful plant viruses. Expression of double‐stranded RNA (dsRNA) directed against suitable insect target genes in transgenic plants has been shown to give protection against pests through plant‐mediated RNA interference (RNAi). Thus, as a potential alternative and effective strategy for insect pest management in agricultural practice, plant‐mediated RNAi for aphid control has received close attention in recent years. In this review, the mechanism of RNAi in insects and the so far explored effective RNAi target genes in aphids, their potential applications in the development of transgenic plants for aphid control and the major challenges in this regard are reviewed, and the future prospects of using plant‐mediated RNAi for aphid control are discussed. This review is intended to be a helpful insight into the generation of aphid‐resistant plants through plant‐mediated RNAi strategy. © 2016 Society of Chemical Industry 相似文献
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Julia Bally Elane Fishilevich Andrew J Bowling Heather E Pence Kenneth E Narva Peter M Waterhouse 《Pest management science》2018,74(8):1751-1758
RNA interference (RNAi) was discovered almost 20 years ago and has been exploited worldwide to silence genes in plants and animals. A decade later, it was found that transforming plants with an RNAi construct targeting an insect gene could protect the plant against feeding by that insect. Production of double‐stranded RNA (dsRNA) in a plant to affect the viability of a herbivorous animal is termed trans‐kingdom RNAi (TK‐RNAi). Since this pioneering work, there have been many further examples of successful TK‐RNAi, but also reports of failed attempts and unrepeatable experiments. Recently, three laboratories have shown that producing dsRNA in a plant's chloroplast, rather than in its cellular cytoplasm, is a very effective way of delivering TK‐RNAi. Our review examines this potentially game‐changing approach and compares it with other transgenic insect‐proofing schemes. © 2018 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry. 相似文献
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Wei-Wen Chen Hong Zhang Yong Chen Wen-Hui Zeng Zhi-Qiang Li 《Pest management science》2023,79(7):2299-2310
BACKGROUND
RNA interference (RNAi) technology is an environmentally friendly strategy for controlling insect pests. Lipopolysaccharide-binding protein (LBP) recognizes lipopolysaccharides, which are a major outer membrane constituent of Gram-negative bacteria. We propose that the LBP gene is a potential target for termite management; however, to date, no studies have examined this gene in termites.RESULTS
In this study, we cloned the LBP gene of Coptotermes formosanus (Cf) and found that the mortality rate of termite workers significantly increased, and the repellence of these workers to Gram-negative bacteria was suppressed after knockdown of CfLBP using double-stranded RNA (dsRNA) injection and feeding. Moreover, the mortality rate of termite workers fed with CfLBP dsRNA and three Gram-negative bacteria (provided separately) was over 50%, which was much higher than that of termites treated with either CfLBP dsRNA or Gram-negative bacteria. Finally, we found that CfLBP impacts the IMD pathway to regulate the immune response of C. formosanus to Gram-negative bacteria.CONCLUSION
CfLBP plays a important role in the immune defense of termites against Gram-negative bacteria. It can be used as an immunosuppressant for RNAi-based termite management and is an ideal target for termite control based on the combined use of RNAi and pathogenic bacteria. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry. 相似文献5.
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BACKGROUND: An efficient and convenient RNA interference (RNAi) technique involving double‐stranded RNA (dsRNA) ingestion is useful for gene function studies of non‐model insects. RESULTS: Three dsRNAs targeting different sites within a gene encoding vacuolar ATP synthase subunit E (V‐ATPase‐E, 21E01) were synthesised for RNAi in Nilaparvata lugens. dsRNA was found to be stable in 0.1 g mL?1 sucrose solution, but unstable in artificial fodder. Therefore, dsRNAs were orally delivered into N. lugens in 0.1 g mL?1 sucrose solution. RNAi was induced by all three of the dsRNAs at 0.05 µg µL?1 in N. lugens. Time dynamics analysis of gene silencing indicated that significant suppression of the target gene began as early as 2 days after ingestion of ds2‐21E01 and ds3‐21E01. However, significant repressive effects were recorded up to 10 days after exposure to ds1‐21E01. The maximum reduction in target gene mRNA was observed after 10 days of treatment, with suppression ratios induced by ds1‐21E01, ds2‐21E01 and ds3‐21E01 of 41, 55 and 48% respectively. CONCLUSION: An efficient and convenient RNAi technique involving dsRNA ingestion has been successfully developed for N. lugens. This will be a useful tool for further functional genomic investigation in this organism. Copyright © 2011 Society of Chemical Industry 相似文献
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Nitish Kunte Erin McGraw Sydney Bell David Held Luz‐Adriana Avila 《Pest management science》2020,76(1):26-41
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 相似文献
8.
Anastasia MW Cooper Kristopher Silver Jianzhen Zhang Yoonseong Park Kun Yan Zhu 《Pest management science》2019,75(1):18-28
RNA interference (RNAi) is an endogenous, sequence‐specific gene‐silencing mechanism elicited by small RNA molecules. RNAi is a powerful reverse genetic tool, and is currently being utilized for managing insects and viruses. Widespread implementation of RNAi‐based pest management strategies is currently hindered by inefficient and highly variable results when different insect species, strains, developmental stages, tissues, and genes are targeted. Mechanistic studies have shown that double‐stranded ribonucleases (dsRNases), endosomal entrapment, deficient function of the core machinery, and inadequate immune stimulation contribute to limited RNAi efficiency. However, a comprehensive understanding of the molecular mechanisms limiting RNAi efficiency remains elusive. Recent advances in dsRNA stability in physiological tissues, dsRNA internalization into cells, the composition and function of the core RNAi machinery, as well as small‐interfering RNA/double‐stranded RNA amplification and spreading mechanisms are reviewed to establish a global understanding of the obstacles impeding wider understanding of RNAi mechanisms in insects. © 2018 Society of Chemical Industry 相似文献
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RNA interference technology in crop protection against arthropod pests,pathogens and nematodes 
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下载免费PDF全文 Moises Zotti Ericmar Avila dos Santos Deise Cagliari Olivier Christiaens Clauvis Nji Tizi Taning Guy Smagghe 《Pest management science》2018,74(6):1239-1250
Scientists have made significant progress in understanding and unraveling several aspects of double‐stranded RNA (dsRNA)‐mediated gene silencing during the last two decades. Now that the RNA interference (RNAi) mechanism is well understood, it is time to consider how to apply the acquired knowledge to agriculture and crop protection. Some RNAi‐based products are already available for farmers and more are expected to reach the market soon. Tailor‐made dsRNA as an active ingredient for biopesticide formulations is considered a raw material that can be used for diverse purposes, from pest control and bee protection against viruses to pesticide resistance management. The RNAi mechanism works at the messenger RNA (mRNA) level, exploiting a sequence‐dependent mode of action, which makes it unique in potency and selectivity compared with conventional agrochemicals. Furthermore, the use of RNAi in crop protection can be achieved by employing plant‐incorporated protectants through plant transformation, but also by non‐transformative strategies such as the use of formulations of sprayable RNAs as direct control agents, resistance factor repressors or developmental disruptors. In this review, RNAi is presented in an agricultural context (discussing products that have been launched on the market or will soon be available), and we go beyond the classical presentation of successful examples of RNAi in pest‐insect control and comprehensively explore its potential for the control of plant pathogens, nematodes and mites, and to fight against diseases and parasites in beneficial insects. Moreover, we also discuss its use as a repressor for the management of pesticide‐resistant weeds and insects. Finally, this review reports on the advances in non‐transformative dsRNA delivery and the production costs of dsRNA, and discusses environmental considerations. © 2017 Society of Chemical Industry 相似文献
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为筛选防控豌豆蚜Acyrthosiphon pisum的关键RNA干扰(RNA interference,RNAi)靶标基因,通过构建精氨酸激酶(arginine kinase,AK)、超氧化物歧化酶(superoxide dismutase,SOD)和几丁质合成酶(chitin synthase,CHS)这3个与豌豆蚜生长发育相关基因的dsRNA,于室内检测ds AK、ds SOD和ds CHS处理后对其致死效果,并比较4种dsRNA递送方式对其致死效果的影响,筛选获得防治效果明显的dsRNA及其递送方式。结果显示,处理48～168 h后,不同基因dsRNA处理组豌豆蚜的死亡率比阴性对照组高2.19倍～4.39倍,其中ds AK处理组豌豆蚜的死亡率为27.7%～91.1%,显著高于ds SOD、ds CHS处理组及阴性对照组。使用ds AK通过人工饲喂法和叶片注射饲喂法处理豌豆蚜,48 h即可出现显著致死效果。ds AK通过人工饲喂方法处理72 h后,豌豆蚜AK基因表达量比空白对照组降低43.2%,同时可抑制豌豆蚜的生长发育,使其体长比阴性对照组减少了5.9%。表明ds AK对豌豆蚜... 相似文献
11.
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。 相似文献
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RNAi (RNA干扰) 是指由诱导分子siRNA (小干扰 RNA)、miRNA (微小RNA)或piRNA (P 转座子诱导互作 RNA) 特异性降解或者抑制同源mRNA,引起靶标基因沉默的现象。RNAi技术具有操作简便、特异性和选择性强等显著特点,是目前农业生命科学领域最有可能应用于病虫害防控的新技术之一。本文通过综述近年来RNAi在农业病虫害防控领域应用的最新研究成果,并对RNAi技术在新靶标基因筛选、高效dsRNA载体开发、与传统农药相结合以及拓宽应用范围等诸多方面的发展前景进行了展望,同时还针对RNAi干扰效率、稳定性、成本控制、抗性发展及抗性治理等方面所面临的挑战进行了深入探讨,提出了合理建议。基于RNAi技术的病虫害防控策略将继续焕发新的活力,为综合防控提供新理念。 相似文献
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Anil Kumar Atul Kakrana Anil Sirohi K. Subramaniam R. Srinivasan M. Z. Abdin Pradeep K. Jain 《Journal of General Plant Pathology》2017,83(2):91-97
Root-knot nematodes (RKNs) are one of the most important biotic factors limiting crop productivity in many crop plants. The major RKN control strategies include development of resistant cultivars, application of nematicides and crop rotation, but each has its own limitations. In recent years, RNA interference (RNAi) has become a powerful approach for developing nematode resistance. The two housekeeping genes, splicing factor and integrase, of Meloidogyne incognita were targeted for engineering nematode resistance using a host-delivered RNAi (HD-RNAi) approach. Splicing factor and integrase genes are essential for nematode development as they are involved in RNA metabolism. Stable homozygous transgenic Arabidopsis lines expressing dsRNA for both genes were generated. In RNAi lines of splicing factor gene, the number of galls, females and egg masses was reduced by 71.4, 74.5 and 86.6%, respectively, as compared with the empty vector controls. Similarly, in RNAi lines of the integrase gene, the number of galls, females and egg masses was reduced up to 59.5, 66.8 and 63.4%, respectively, compared with the empty vector controls. Expression analysis revealed a reduction in mRNA abundance of both targeted genes in female nematodes feeding on transgenic plants expressing dsRNA constructs. The silencing of housekeeping genes in the nematodes through HD-RNAi significantly reduced root-knot nematode infectivity and suggests that they will be useful in developing RKN resistance in crop plants. 相似文献
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草地贪夜蛾Spodoptera frugiperda是一种世界性重大农业害虫,在全球多个国家普遍发生,其幼虫可为害玉米、水稻等多种农作物。该虫于2019年初入侵我国,对我国农业生产构成了严重的威胁,防控形势严峻。为寻求一种草地贪夜蛾的绿色防控方法,本文对草地贪夜蛾潜在RNA干扰(RNA interference,RNAi)靶标致死基因、RNAi传统双链RNA(double-stranded RNA,dsRNA)递送技术的瓶颈以及纳米载体介导的RNAi技术应用进行概括,并对纳米载体介导的RNAi技术应用前景进行展望。 相似文献
15.
喷施诱导的基因沉默(spray-induced gene silencing, SIGS)技术是一种新型基因沉默技术, 其以病原菌生长发育和致病相关基因为靶标, 将体外合成的针对靶基因的dsRNA喷施于植物表面, 抑制靶基因的表达?在病原菌侵染寄主的过程中, 病原菌可直接从寄主植物表面摄取dsRNA, 也可由植物吸收dsRNA后, 直接以dsRNA的形式或将dsRNA剪切成sRNAs转运至病原菌体内诱导病原菌相关基因沉默, 从而抑制病菌的侵染和扩展?由于SIGS技术不需要转基因, 再加上其具有高效性?特异性和环境友好性, 故显示出巨大的应用潜力?本文综述了SIGS技术的最新研究进展, 总结了影响SIGS控制植物病害的因素及未来的研究方向, 展望了其应用前景? 相似文献
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RNAi as an emerging approach to control Fusarium head blight disease and mycotoxin contamination in cereals 下载免费PDF全文
下载免费PDF全文 Ana Karla Machado Neil A Brown Martin Urban Kostya Kanyuka Kim E Hammond‐Kosack 《Pest management science》2018,74(4):790-799
Fusarium graminearum is a major fungal pathogen of cereals worldwide, causing seedling, stem base and floral diseases, including Fusarium head blight (FHB). In addition to yield and quality losses, FHB contaminates cereal grain with mycotoxins, including deoxynivalenol, which are harmful to human, animal and ecosystem health. Currently, FHB control is only partially effective due to several intractable problems. RNA interference (RNAi) is a natural mechanism that regulates gene expression. RNAi has been exploited in the development of new genomic tools that allow the targeted silencing of genes of interest in many eukaryotes. Host‐induced gene silencing (HIGS) is a transgenic technology used to silence fungal genes in planta during attempted infection and thereby reduces disease levels. HIGS relies on the host plant's ability to produce mobile small interfering RNA molecules, generated from long double‐stranded RNA, which are complementary to targeted fungal genes. These molecules are transferred from the plant to invading fungi via an uncharacterised mechanism, to cause gene silencing. Here, we describe recent advances in RNAi‐mediated control of plant pathogenic fungi, highlighting the key advantages and disadvantages. We then discuss the developments and implications of combining HIGS with other methods of disease control. © 2017 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry. 相似文献
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RNAi (RNA interference) 是一种由dsRNA参与、对靶基因表达进行干扰或沉默的现象。由此发展起来的RNAi基因沉默技术已成为当今植物基因功能研究和遗传改良的一个重要手段。该技术已经在靶向病原物(真菌、细菌、病毒和线虫)基因沉默方面得到了广泛的应用,并且产生了一批抗病性增强的转基因植物。人工设计和合成的amiRNAs和ata siRNAs的成功研发加快了RNAi技术的应用。本文对RNAi基因沉默机制、RNAi技术研发进展及其在植物抗病性遗传改良中的应用进行综述,并对其应用策略进行探讨。 相似文献
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Gijs A Kleter 《Pest management science》2020,76(10):3333-3339
Genetically modified crops have been grown commercially for more than two decades. Some of these crops have been modified with genetic constructs that induce gene silencing through RNA interference (RNAi). The targets for this silencing action are genes, either specific endogenous ones of the host plant or those of particular pests or pathogens infesting these plants. Recently emerging new genetic tools enable precise DNA edits with the same silencing effect and have also increased our knowledge and insights into the mechanisms of RNAi. For the assessment of the safety of foodstuffs from crops modified with RNAi, internationally harmonized principles for risk assessment of foods derived from genetically modified crops can be followed. Special considerations may apply to the newly expressed silencing RNA molecules, such as their possible uptake by consumers and interference with expression of host genes, which, however, would need to overcome many barriers. Bioinformatics tools aid the prediction of possible interference by a given RNA molecule with the expression of genes with homologous sequences in the host crop and in other organisms, or possible off‐target edits in gene‐edited crops. © 2020 The Author. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry. 相似文献
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BACKGROUND: Dose–response relationships for antifeedant effects of H. lupulus extracts against larvae and adults of the Colorado potato beetle (CPB) were determined in laboratory conditions. The larval and adult beetles were fed on potato leaflets treated with H. lupulus extract ranging from 0.4 to 40 mg mL?1 in a no‐choice situation. Their feeding behavior was recorded, and larval growth and antifeedant indexes were calculated. RESULTS: H. lupulus treatments significantly affected larval growth rate, and at higher concentrations the larval weights were significantly reduced over the course of the assay. Adults of CPB were more sensitive to the extracts than the larvae, and, even at lower doses, adult beetles were arrested for longer periods than larvae. CONCLUSION: These results indicate that H. lupulus extracts may have potential for control of CPB, particularly in organic farms where conventional insecticides are not available. Copyright © 2012 Society of Chemical Industry 相似文献