二化螟对双酰胺类杀虫剂抗性的分子机制研究进展

新型双酰胺类杀虫剂已广泛用于保障水稻生产,而二化螟作为危害水稻生产的钻蛀性害虫,已经对该类杀虫剂产生了抗性,明确该类杀虫剂抗性分子机制,可为二化螟抗性快速检测和绿色防控提供技术支撑。本文在总结二化螟对双酰胺类杀虫剂抗性现状的基础上,重点综述了近年来有关其抗性分子机制研究的进展,主要包括解毒酶和转运蛋白基因过表达介导的代谢抗性,以及鱼尼丁受体基因突变介导的靶标抗性;指出了该研究领域在抗性分子检测、抗性新基因鉴定、抗性基因调控网络和多重抗性机制等方面存在的问题,并展望了其发展方向,认为：利用高通量测序技术检测害虫种群抗药性;利用多组学技术鉴定新抗性基因及调控网络,以探明多重抗性机制;将反向遗传学工具放射性配基结合及电生理技术深入验证抗性基因功能;需开发靶向抗性基因的dsRNA转基因作物、纳米农药及选择性新型化学杀虫剂,以达到杀虫剂减施增效的目的。     

棉蚜抗药性现状及治理策略

棉蚜Aphis gossypii Glover是农业生产上最重要的害虫之一。化学杀虫剂一直以来都是棉蚜综合防治体系中的重要组成部分, 但化学杀虫剂的不合理使用导致棉蚜对多种杀虫剂均产生了高水平抗性。现有研究表明, 靶标位点突变、解毒酶基因的过表达以及某些肠道共生菌丰度的变化是导致棉蚜对杀虫剂产生抗性的主要机制。针对棉蚜抗性发展现状及其抗药性机制, 制定科学合理的抗性治理策略, 是充分发挥化学防治的优势、实现棉蚜可持续治理的关键。本文主要从棉蚜的抗药性现状、抗性机制和抗性治理策略3个方面对近10多年的主要进展进行了综述, 旨在为棉蚜抗药性长效治理和科学施策提供理论依据。     

害虫对Bt杀虫蛋白抗性相关受体蛋白互作网络分析

为对Bt杀虫蛋白与化学杀虫剂的协同使用提供理论指导，通过蛋白互作分析方法，以黑腹果蝇Drosophila melanogaster为模型，对化学杀虫剂与Bt杀虫剂作用受体以及抗性相关蛋白的互作网络进行研究。结果表明，Bt杀虫蛋白的27个受体在黑腹果蝇中共存在50个互作蛋白，其中有39个蛋白只与Bt杀虫蛋白抗性相关，11个蛋白还与化学杀虫剂抗性相关，这2类杀虫剂抗性互作网络交集较小，并且重叠蛋白的变异引起交互抗性的概率不大。表明Bt杀虫蛋白与化学杀虫剂产生交互抗性的概率较小，可以将Bt杀虫剂与化学杀虫剂协同使用，提高害虫防治效果，并有效克服或延缓害虫抗性产生。     

Genetically engineering better fungal biopesticides

Microbial insect pathogens offer an alternative means of pest control with the potential to wean us off our heavy reliance on chemical pesticides. Insect pathogenic fungi play an important natural role in controlling disease vectors and agricultural pests. Most commercial products employ Ascomycetes in the genera Metarhizium and Beauveria. However, their utilization has been limited by inconsistent field results as a consequence of sensitivity to abiotic stresses and naturally low virulence. Other naturally occurring biocontrol agents also face these hurdles to successful application, but the availability of complete genomes and recombinant DNA technologies have facilitated design of multiple fungal pathogens with enhanced virulence and stress resistance. Many natural and synthetic genes have been inserted into entomopathogen genomes. Some of the biggest gains in virulence have been obtained using genes encoding neurotoxic peptides, peptides that manipulate host physiology and proteases and chitinases that degrade the insect cuticle. Prokaryotes, particularly extremophiles, are useful sources of genes for improving entomopathogen resistance to ultraviolet (UV) radiation. These biological insecticides are environmentally friendly and cost‐effective insect pest control options. © 2017 Society of Chemical Industry     

昆虫对拟除虫菊酯类杀虫剂的代谢抗性机制研究进展

随着拟除虫菊酯类杀虫剂在卫生和农业害虫防治中的广泛应用,昆虫对此类杀虫剂产生抗性的报道越来越多。目前已明确昆虫对拟除虫菊酯类杀虫剂的抗性机制包括表皮穿透率下降、靶标抗性以及代谢抗性,其中代谢抗性机制较为普遍,而且其与昆虫对多种杀虫剂的交互抗性关系密切。目前,随着基因组、转录组以及蛋白质组学等新技术的发展及应用,昆虫对拟除虫菊酯类杀虫剂的代谢抗性机制研究也取得了很多新进展。昆虫体内细胞色素P450酶（P450s）、羧酸酯酶（CarE）及谷胱甘肽S-转移酶（GSTs）等重要解毒酶系的改变均与昆虫对拟除虫菊酯类杀虫剂的代谢抗性有关,其中这3类解毒酶的活性及相关基因表达量的变化是昆虫对此类杀虫剂产生代谢抗性的主要原因。明确昆虫对拟除虫菊酯类杀虫剂的代谢抗性机制,对合理使用此类杀虫剂及延缓抗药性的产生均具有重要意义。本文在总结拟除虫菊酯类杀虫剂代谢路径及相关生物酶研究概况的基础上,综述了近年来有关昆虫对此类杀虫剂代谢抗性机制研究的主要进展。     

Review: Resistance as a concomitant of modern crop protection

This paper reviews the impact of resistance to fungicides and insecticides/acaricides on the way crop protection is practised. It is now clear that resistance can develop to virtually any crop-protection product, in any pest, fungal pathogen or even weed. As a limiting factor in crop protection, it is a fact of life. A positive side-effect is the precision with which products are used today, with increasing implementation of Integrated Pest Management (IPM) programmes. This is a vital step towards sustainability. This paper describes: past experiences; current status of resistance; how resistance management influences current crop protection practices; regulatory aspects; and the outlook for the future. It concludes that EU regulations on resistance management must be simple and workable. Chemicals will continue to have a central role in optimising yields from the world's crops, as new tools, including biotechnology, become available for crop protection and resistance management. The crop-protection industry's innovations and product stewardship programmes will contribute to sustainable agriculture. This will provide continued benefits to users, the environment and society. ©1997 SCI     

Resistance and cross-resistance to the spinosyns - A review and analysis

The spinosyns were introduced in 1997 with the launch of spinosad. Since then, there have been several cases of resistance to spinosad in the field populations of insect pest species that have resulted in reduced efficacy. There have also been a number of studies where spinosad-resistant insect strains were created and characterized in the laboratory. Likewise many studies examining resistance to a variety of other classes of insecticides have included spinosad in the evaluation of their cross-resistance spectrum. Understanding mechanisms of resistance and cross-resistance can provide the basis for developing insecticide resistance management programs, as well as defining the most appropriate tools to address potential resistance issues. This review provides an overview and analysis of resistance and cross-resistance to the spinosyns (spinosad and spinetoram). Although there are more than 30 examples of resistance to the spinosyns, only half of these are related to selection in the field. The majority of these field selected examples occur in either the diamondback moth or western flower thrips. There have also been over 85 studies investigating cross-resistance to the spinosyns. However, in half of these studies spinosad showed no cross-resistance, and in another third of the total studies spinosad cross-resistance was minimal. Therefore, while resistance and cross-resistance to the spinosyns does occur, it is appears to be more limited in impact than might be implied from the large body of literature on the subject.     

The Molecular Biology of the Interactions Between Trichoderma spp., Phytopathogenic Fungi, and Plants

ABSTRACT Trichoderma-based biofungicides are a reality in agriculture, with more than 50 formulations today available as registered products worldwide. Several strategies have been applied to identify the main genes and compounds involved in this complex, three-way cross-talk between the fungal antagonist, the plant, and microbial pathogens. Proteome and genome analysis have greatly enhanced our ability to conduct holistic and genome-based functional studies. We have identified and determined the role of a variety of novel genes and gene-products, including ABC transporters, enzymes and other proteins that produce or act as novel elicitors of induced resistance, proteins responsible for a gene-for-gene avirulent interaction between Trichoderma spp. and plants, mycoparasitism-related inducers, plant proteins specifically induced by Trichoderma, etc. We have transgenically demonstrated the ability of Trichoderma spp. to transfer heterologous proteins into plant during root colonization, and have used green fluorescent protein and other markers to study the interaction in vivo and in situ between Trichoderma spp. and the fungal pathogen or the plant.     

真菌防治蝗虫研究进展

蝗虫是世界性重大成灾害虫,时常在草原、沙漠区或农作区暴发为害.当前的非洲沙漠蝗灾害再次提醒人们,蝗灾依然是粮食生产和生态安全的严重威胁.在寻求绿色可持续防蝗的新策略新技术研究中,昆虫病原真菌显现出种类多、毒力强、易规模化生产及环境友好等明显优势而倍受重视.近30年,真菌防蝗研究和应用进步加快,国际、国内都有真菌防蝗产品...     

Biochemistry of insecticide resistance

After briefly summarizing various resistance mechanisms and their effects on certain types of insecticides, attention is focused on gaps in the understanding of resistance. Most resistance mechanisms consist either of a change in sensitivity of the site of action or an increased capacity for detoxication. In both cases such changes could theoretically be due to quantitative changes of an enzyme (more sites of action or more detoxication enzyme) or to qualitative changes (less-sensitive site of action or more efficient detoxication enzyme). Examples have now been found for each of these possible causes of resistance to insecticides, except for an increased amount of sites of action. Whereas examples of the latter mechanism are known for cell lines resistant to anti-cancer chemicals, so far they have not been found in resistance to insecticides. The impact of the different mechanisms on development and stability of resistance is discussed.     

解毒酶和转运蛋白介导的害虫抗药性分子机制研究进展

害虫抗药性是导致杀虫剂防效降低的一个重要因素,而抗性机制的阐明是害虫抗药性综合治理的基础。研究表明,代谢能力增强是害虫抗药性产生的重要原因,害虫对杀虫剂等外源物质的代谢需要细胞色素P450酶系(P450s)、羧酸酯酶(CarEs)、谷胱甘肽S-转移酶(GSTs)、UDP-葡萄糖醛酸转移酶(UGTs)和ATP结合盒转运蛋白(简称ABC转运蛋白)等解毒酶和转运蛋白的参与。结合近年来对害虫抗药性分子机制的研究进展,本文综述了上述解毒酶和转运蛋白参与杀虫剂抗药性的分子机制,并对害虫抗药性治理的新方法进行了展望。     

昆虫对双酰胺类杀虫剂抗性机制研究进展

双酰胺类杀虫剂是以昆虫鱼尼丁受体为作用靶标的新型杀虫剂,由于其作用机制独特,对多种鳞翅目害虫具有良好的防治效果而得到广泛应用。但已经有多种害虫的田间种群对该类药剂产生了抗性,甚至导致田间防治失败。本文在综述昆虫对双酰胺类杀虫剂抗性现状的基础上,重点总结了抗性机制方面的最新研究进展,并对今后的研究方向进行了展望,以期为进一步深入研究双酰胺类杀虫剂的抗性机制提供借鉴。     

灰飞虱对杀虫剂的抗性分子机制研究进展

灰飞虱是中国长江流域和黄淮地区重要的农业害虫,由于杀虫剂的广泛与大量使用,已导致其对多种杀虫剂产生了抗性。深入研究其抗药性分子机制,可为灰飞虱抗性的快速检测和治理提供重要理论基础。文章总结了灰飞虱对毒死蜱、吡虫啉、溴氰菊酯、噻嗪酮、氟虫腈和乙虫腈等杀虫剂的抗性分子机制研究进展,主要包括抗性相关解毒酶和转运蛋白基因的筛选与功能验证,以及靶标位点突变等重要研究成果,指出该研究领域当前存在的问题主要有抗性基因的功能验证及调控路径、抗性新基因的鉴定及交互抗性和多重抗性机制不明确等,并展望了其未来发展方向,认为:可利用CRISPR/Cas9基因编辑技术验证抗性基因功能;可将转录组测序结合生物信息学手段用于鉴定新抗性基因及抗性调控基因,以探明交互抗性和多重抗性机制;应深入至蛋白组学水平探讨抗性机制;需开发配套的高效田间施药技术,以达到杀虫剂减施增效的目的。     

Strategies for the prevention of development of pesticide resistance in the UK—lessons for and from the use of herbicides,fungicides and insecticides

Since the 1950s, pesticide resistance has been identified in many species. This paper considers the role of resistance action groups and notes that they were all formed in response to resistance problems occurring. Data now exist on the strategies which are most effective and the paper aims to bring together information from the fields of weeds, pests and diseases. Pesticide mixtures, sequences or rotations have been demonstrated as having a clear role in resistance management strategies. Resistance management would be improved if there was agreement on uniform test methodology and interpretation of results. The industry must work together to agree what constitutes an anti-resistance strategy, whether this is for prevention or cure, and to ensure that this is then included within regulatory frameworks. Future developments such as patch treatment, biotechnology and biocontrol are discussed. It is concluded that, to date, there has been little discussion between specialists in the field of resistance to herbicides, fungicides or insecticides and it is clear there are significant advantages to be had from more interaction. ©1997 SCI     

Purification and characterisation of proteins secreted by the entomopathogenic fungus Metarhizium anisopliae with insecticidal activity against adults of the Mediterranean fruit fly,Ceratitis capitata (Diptera: Tephritidae)

BACKGROUND: The control of the Mediterranean fruit fly (medfly) Ceratitis capitata (Wied) is usually performed with protein bait sprays incorporating chemical insecticides that may have adverse effects on humans, non‐target organisms and the environment. In recent years, scientists have sought more environmentally friendly insecticides for medfly control, such as plant‐ and microorganism‐derived compounds. Among these compounds, entomopathogenic fungi are an unexplored source of natural insecticides. RESULTS: The crude soluble protein extract (CSPE) of the entomopathogenic fungus Metarhizium anisopliae (Mestch.) (strain EAMa 01/58‐Su) shows chronic insecticidal activity when administered per os. Mortality in flies exhibits a dose response. The CSPE produces an antifeedant effect in adult flies, a result probably due to a progressive deterioration of the fly midgut after ingestion of the extract. Protease and temperature treatments show that insecticidal activity against C. capitata is due to proteinaceous compounds that are highly thermostable. Four monomeric proteins from this crude extract have been purified by liquid chromatography and gel electroelution. Although all four monomers seem to be involved in the insecticidal activity of the CSPE, the 15 kDa and the 11 kDa proteins appear to be mainly responsible for the observed insecticidal effect. CONCLUSIONS: Four new fungal proteins with insecticidal activity have been purified and identified. These proteins might be combined with insect baits for C. capitata biocontrol. Copyright © 2009 Society of Chemical Industry     

Can selective peptides be combined efficiently with agrochemicals? A new approach to insect control

Sodium channels have been a major target for the development of insecticides such as synthetic pyrethroids. However, insecticides currently available induce resistance and present limited selectivity to insect pests. Molecular and biochemical studies, as well as binding experiments using radiolabelled neurotoxins, have shown that sodium channels expressed in various insect orders must be structurally and pharmacologically different. At least three groups of peptide neurotoxins derived from scorpion venom are highly active on insects and very weakly or practically inactive on mammals. It is proposed that various insecticides are examined for possible cooperative interactions with the peptide toxins highly active on insects, and pairs of ligands are identified that will increase the selectivity not only between mammals and insects but also between different pest and non-pest insects. This is feasible on the basis of the differential allosteric modulations observed between LqhαIT, an α-toxin highly active on insects, and brevetoxin on locust versus cockroach and rat brain sodium channels. Moreover, combination of LqhαIT with the pyrethroid deltamethrin increased the binding of [¹²⁵I] LqhαIT by more than 1.8-fold, and the combined presence of brevetoxin further increased the binding. Such allosteric modulation may provide a new approach to increase the selective activity of pesticides on target organisms by simultaneous application of allosterically interacting drugs, designed on the basis of the selective peptide toxins.     

Culex pipiens pallens: identification of genes differentially expressed in deltamethrin-resistant and -susceptible strains

Insecticide-resistance is a major obstacle to controlling insect vectors of microorganisms that cause human diseases. Identification of genes associated with resistance to insecticides has been a valuable tool for understanding mechanisms underlying resistance to commonly used insecticides such as deltamethrin. To identify such genes, we used suppression subtractive hybridization to obtain 809 differentially expressed clones in deltamethrin resistant versus susceptible laboratory strains of Culex pipiens pallens. Using cDNA microarrays and reverse Northern blots, a subset of 16 clones was confirmed to have greater than 3-fold difference in expression levels. Within this subset, we identified 2 clones uniquely expressed in the deltamethrin-resistant strain, eight clones exhibiting higher expression in the resistant strain and six in the susceptible strain. Of these 16 clones, 13 clones have sequence homology to known genes, such as ribosomal RNA, ribosome proteins, trypsin, and chymotrypsin-like proteins. Our data suggests resistance to deltamethrin may be a polygenic phenotype.     

细菌生物被膜在生物防治中的作用

细菌生物被膜（bacterial biofilm,BBF）是在固—液和气—液表面上生长并封闭在胞外多糖基质中的细胞群,能调控细菌对环境胁迫的适应能力,提高细菌对紫外线（UV）胁迫等环境的抗逆性,是细菌的一种保护性生长方式。生防菌剂（biological control agents,BCAs）因其能以环境友好的方式长期控制病害虫等优点,一直是生物农药领域研究的热点。本文简介了细菌生物被膜及其应用,以苏云金芽胞杆菌（Bacillus thuringiensis,Bt）和枯草芽胞杆菌（Bacillus subtilis,Bs）为核心综述了BBF对生防活性的影响、生防细菌生物被膜形成、解离的表达、调控机制以及生物被膜调控基因的鉴定方法,并指出未来BBF在生防应用方面主要的研究方向,旨在为解决BCAs田间持效期较短等问题提供新的思路。     

Knockdown resistance to DDT and pyrethroids: from target-site mutations to molecular modelling

Naturally derived insecticides such as pyrethrum and man-made insecticides such as DDT and the synthetic pyrethroids act on the voltage-gated sodium channel proteins found in insect nerve-cell membranes. The correct functioning of these channels is essential for the normal transmission of nerve impulses, and this process is disrupted by binding of the insecticides, leading to paralysis and eventual death. Some insect pest populations have evolved modifications of the sodium channel protein that inhibit the binding of the insecticide and result in the insect developing resistance. This perspective outlines the current understanding of the molecular processes underlying target-site resistance to these insecticides (termed kdr and super-kdr), and how this knowledge may in future contribute to the design of novel insecticidal compounds.     

我国部分地区鲜食辣椒及其加工品中辣椒轻斑驳病毒的调查

本研究利用RT-PCR技术对我国部分地区鲜食辣椒和辣椒加工品中的辣椒轻斑驳病毒Pepper mild mottle virus(PMMoV)进行了调查。其中,鲜食辣椒样品(249份)中PMMoV检出率为75.5%;147份辣椒加工品中有97份检测到PMMoV,且所采集的每一类别的辣椒制品中都有阳性样品。调查结果表明:PMMoV不仅广泛分布于我国辣椒种植区,而且在多种辣椒加工品中的检出率也较高。该研究进一步明确了PMMoV的来源及可能的传播途径,为该病毒的防控提供了理论依据。