小菜蛾对杀虫剂抗性的研究：Ⅱ.小菜蛾幼虫的酯酶分析

本文研究了酯酶在小菜蛾抗药性中的作用。离体酯酶活力测定表明，抗性品系幼虫的羧酸酯酶活力是敏感品系的１．４倍，故羧酸酯酶在抗性品系抗药性的作用不大，酸性和碱性磷酸酯酶在抗性和敏感品系的幼虫中差异很小，幼虫的酸性和碱性磷酸酯酶电泳酶谱没有差异。幼虫酯酶同工酶电泳分析显示Ｅｓｔ－１位点的Ｃ基因的频率随着抗性水平的提高而增加，基因型的多态性提高。     

小菜蛾对杀虫药剂抗性的研究 Ⅱ．小菜蛾幼虫的酯酶分析

本文研究了酯酶在小菜蛾Ｐｌｕｔｅｌａｘｙｌｏｓｔｅｌａ抗药性中的作用。离体酯酶活力测定表明，抗性品系（ＧＺ）幼虫的羧酸酯酶活力是敏感品系（ＳＳ）的１．４倍，故羧酸酯酶在抗性品系（ＧＺ）抗药性的作用不大；酸性和碱性磷酸酯酶在抗性和敏感品系的幼虫中差异很小，幼虫的酸性和碱性磷酸酯酶电泳酶谱没有差异。幼虫酯酶同工酶电泳分析显示Ｅｓｔ－１位点的Ｃ基因的频率随着抗性水平的提高而增加，基因型的多态性也提高。     

北京及河北省北部瓜-棉蚜对拟除虫菊酯抗药性的研究初报

1983—1986年就北京及河北省廊坊、香河、涿鹿等地区的瓜-棉蚜(Aphis gossypii Glov.)对拟除虫菊酯杀虫药剂的抗药性作了系统监测。试验结果表明,北京、廊坊及香河等地区的瓜-棉蚜种群对氰戊菊酯及溴氰菊酯已产生了一定程度的抗性。以LC50及LC95进行抗性比较,对氰戊菊酯分别为519和6416倍;对溴氰菊酯为401和14805倍。蚜体中水解α-乙酸萘酯酶(以下简称酯酶)的活性与其对氰戊菊酯及溴氰菊酯的抗性程度密切相关,相关系数均大于0.9,均在显著水平以上。因此、酯酶是瓜-棉蚜抗拟除虫菊酯的主要抗性机制之一。     

不同寄主植物棉蚜酯酶对杀虫剂不敏感性的频率分布

通过生物测定和生化分析研究了花椒棉蚜(HA)和棉花棉蚜(MA)种群对杀虫剂的抗性及其α-乙酸萘酯(α-NA)酯酶和乙酰胆碱酯酶(AChE)对杀虫剂的不敏感性。结果表明,MA种群对甲基对硫磷、辛硫磷和灭多威的耐药性均高于HA种群;MA种群中具有高α-NA酯酶活力的个体比HA种群多,甲基对硫磷对HA和MA种群α-NA酯酶的抑制率达30％时的个体累计频率分别为37％和65％;抑制率大于50％时的累计频率分别为18％和1％;灭多威对HA种群棉蚜AChE的抑制作用高于MA种群。说明MA种群对有机磷的抗性可能与α-NA酯酶活力的增高和对杀虫剂的不敏感性有关,对灭多威的抗性与AChE敏感度的降低有关。滤纸酯酶反应法和微量滴度酶标板法均能快速、有效地测定棉蚜个体酯酶的频率分布,准确地反映两种群棉蚜酯酶活力的差异。     

介绍一种昆虫抗药性测定的新方法

一般认为昆虫抗药性是与昆虫体内抗性代谢的解毒酶活性增高和乙酰胆咸酯酶(AchE)敏感性降低有关。在进行AchE有关的抗性检测时,经典的方法是“微滴板比色法”,这种方法较为敏感迅速,但需特殊仪器设备,为了适应抗性监测和抗性机理的研究需要,这里介绍一种由美国加州大学D.Dary教授最近报道的斑点法,该实验步聚如下:1 试验昆虫取羽化3天的成虫,家蝇可取成蝇头部,逐个放在滴定瓷板的空穴里。     

聚合酶链反应（PCR）技术在蚊虫抗性基因研究中的应用

应用ＰＣＲ技术对广泛部分地区的致倦库蚊ＯＰ抗性基因进行测定，实验证明广州地区自然种群和实验室株有相当数量致倦库蚊有扩增的抗性酯酶Ｂ１基因存在。对实验室株及四个地区自然群体的抗性基因进行扩增，发现实验室株阳性率比自然群体为高，四个地区自然群体抗性基因频率分别为２２．５％、２０％、１５％、和７．５％。另外发现雄性库蚊出现的扩增阳性率比雌蚊高。所设计的两对引物均能成功的扩增致倦库蚊抗性基因酯酶Ｂ１，但后     

朱砂叶螨抗药性与羧酸酯酶活性的关系

羧酸酯酶是昆虫体内普遍存在的一种水解酶。昆虫对多数有机磷、氨基甲酸酯及有些菊酯类杀虫剂产生抗性羧酸酯酶活性有关[1～2]。桃蚜(Muscapersicae)对有机磷的抗性与其体内所含酯酶活力呈正相关,E4酯酶的量随着抗性程度的增加而呈几何级数上升[3]。棉蚜(Aphisgossypii)对对硫磷和对氧磷产生抗性后,α-乙酸萘酯酶活力分别增加15.25倍和60.30倍[4]。本文研究了朱砂叶螨(Tetranychuscinnabarinus)对氧化乐果(有机磷)、三氯杀螨醇(有机氯)、双甲脒(有机氮)及哒螨灵(杂环类)的抗性与羧酸酯酶的关系。1　材料与方法1.1　供试叶螨在人工气象室内…     

不同抗性水平的家强在不同用药方式下抗性机制的演变

增效作用和生化研究结果表明,对溴氰菊酯具有中等抗性和高等抗性水平的家蝇,在不同用药方式作用下,羧酸酯酶、多功能氧化酶的酶活力都有不同程度的提高,谷胱甘肽-S-转移酶和乙酰胆碱酯酶的活力变化不大;在轮换用药和混合用药方式下,乙酰胆碱酯酶与底物亲和力降低。     

不同抗性水平的家绳在不同用药方式下抗性机制的演变

增效作用和生化研究结果表明 ,对溴氰菊酯具有中等抗性和高等抗性水平的家蝇 ,在不同用药方式作用下 ,羧酸酯酶、多功能氧化酶的酶活力都有不同程度的提高 ,谷胱甘肽 - S-转移酶和乙酰胆碱酯酶的活力变化不大 ;在轮换用药和混合用药方式下 ,乙酰胆碱酯酶与底物亲和力降低。     

二斑叶螨对梅岭霉素和溴虫腈的抗性选育及其解毒酶活力变化

在温室内模拟田间药剂的选择压力,用梅岭霉素和溴虫腈对二斑叶螨Tetranychus urticae以浸渍法逐代处理,选育其抗性种群,评价其抗性风险性.结果表明,选育至12代,对梅岭霉素抗性增长到9.2倍,对溴虫腈抗性增长到10.0倍.酶抑制剂和离体酶活性的测定结果表明,二斑叶螨对梅岭霉素的抗性可能与羧酸酯酶、多功能氧化酶解毒作用的增强和药剂对害螨体壁穿透性降低有关;而药剂对害螨体壁穿透性降低和谷胱甘肽-S-转移酶及酯酶代谢作用的增强可能是二斑叶螨对溴虫腈产生抗性的主要原因.     

Polymerase chain reaction-based monitoring techniques for the detection of insecticide resistance-associated point mutations and their potential applications

The design of three PCR-based monitoring techniques for the genotyping of cyclodiene-resistant insects are described: (1) PCR followed by a diagnostic restriction enzyme digest or PCR/REN, (2) PCR amplification of specific alleles or PASA and (3) single-stranded conformational polymorphism analysis of PCR-amplified DNA or SSCP. The relative disadvantages and potential applications of each of these techniques are discussed and compared to the use of insecticide bioassays. It is concluded that, although such techniques will probably never replace bioassays for routine monitoring, they can more readily address several fundamental questions relating to the evolution and spread of specific resistance alleles in insect populations.     

功能基因组技术在昆虫抗药性研究中的应用展望

目前化学防治仍是植物保护的重要手段,化学杀虫剂通过参与昆虫自然选择,诱导其产生可遗传的基因突变,从而导致昆虫抗药性的增加,近年来随着杀虫剂的广泛使用,抗药性已成为植物保护领域亟需解决的重大问题。利用分子生物学手段对由基因变异引起的昆虫抗药性进行解析已成为可能,在现今大多数昆虫抗药性研究中,功能基因组技术已被广泛用于候选基因调控抗药性机制的研究。本文介绍了双元基因表达技术——GAL4/UAS技术、基因干扰技术——RNAi技术和基因编辑技术——CRISPR/Cas9技术这3种有助于阐明昆虫抗药性分子机制的功能基因组技术,综述了这3种功能基因组技术在近几年昆虫抗药性研究中的应用实例,讨论了其目前的发展状况及其优势和局限性,以期在完善的功能基因组技术支持下为昆虫抗药性研究取得更多突破性进展。     

Western flower thrips resistance to insecticides: detection, mechanisms and management strategies

Insecticide resistance continues to be one of the most important issues facing agricultural production. The challenges in insecticide resistance and its management are exemplified by the situation with the western flower thrips Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae). This highly invasive pest has a great propensity for developing insecticide resistance because of its biological attributes, and cases of resistance to most classes of insecticides used for its management have been detected. To combat insecticide resistance in the western flower thrips, several insecticide resistance management (IRM) programs have been developed around the world, and these are discussed. Successful programs rely on non-insecticidal tactics, such as biological and cultural controls and host plant resistance, to reduce population pressures, rotations among insecticides of different mode of action classes to conserve insecticide efficacy, resistance monitoring, sampling to determine the need for insecticide applications and education to assure proper implementation. More judicious insecticide use is possible with the development of well-founded economic thresholds for more cropping systems. While growers will continue to rely on insecticides as part of western-flower-thrips- and thrips-transmitted virus management, more effective management of these pests will be achieved by considering their management in the context of overall integrated pest management, with IRM being a key component of those comprehensive programs.     

浙江省水稻二化螟抗药性现状与治理对策

二化螟是浙江省水稻的主要害虫。由于连续使用杀虫双超过20a，三唑磷10a防治二化螟，致使二化螟对杀虫单(杀虫双)普遍产生抗药性。1999年测定抗性增加67～587倍，达到高抗和极高抗水平；二化螟对三唑磷在温州和台州等局部地区也达到了68～162倍的高抗水平。为治理二化螟抗药性，必须加强对害虫抗药性的监测，在高抗药性地区停止使用杀虫单(杀虫双)和三唑磷防治二化螟；在低抗药性和敏感地区也应减少使用次数和用量，达到延缓抗药性产生的目的。     

Insecticide resistance patterns among colorado potato beetle,leptinotarsa decemlineata (Say) (Coleoptera: chrysomelidae), populations in north carolina

A residual bioassay was used to determine insecticide susceptibility in populations of Colarado potato beetle, Leptinotarsa decemlineata (Say), in eastern North Carolina. Mean bioassay mortalities were subjected to correlation analysis to assess patterns of resistance among tested insecticides. Within insecticide classes, cross-resistance patterns were observed between carbaryl and carbofuran (carbamates), fenvalerate and permethrin (pyrethroids), and among azinphosmethyl, methamidophos, and phosmet (organophosphates). Significant associations between insecticides from different insecticide classes were concluded to be due to multiple resistance. Knowledge of multiple and cross-resistance patterns are discussed in relation to the bioassay as a tool for monitoring insecticide resistance.     

性信息素诱捕器用作田间棉红铃虫种群抗药性的监测

本文提供一种用性信息素粘胶诱捕器收集田间棉红铃虫[Pectinophora gossypiella(Saun-ders)]种群的取样技术,只需简单的恒温设备,就能快速、简便、正确地监测田间红铃虫种群对氰戊菊酯的抗性。经浙江省4个棉区的监测,证明此方法能明显地反映出各地棉红铃虫种群的抗性程度。在用药水平较高的慈溪,红铃虫对氰戊菊酯的抗性已高达26—28倍;在同一棉区,红铃虫越冬代和第二代雄蛾对氰戊菊酯的抗性水平相似,而第一代较敏感。此方法可用于红铃虫抗性的普查,为红铃虫抗药性的综合治理提供依据。     

Field-simulator study of insecticide resistance conferred by esterase-, MACE- and kdr-based mechanisms in the peach-potato aphid,Myzus persicae (Sulzer)

Insecticide sprays were applied to Myzus persicae (Sulzer) populations carrying various combinations of three insecticide resistance mechanisms (esterase-based metabolic resistance and two target site mechanisms, known as MACE and kdr), supported on host plants growing in field simulator cages. The study showed that MACE confers extreme resistance to pirimicarb and triazamate (carbamate insecticides) but not to deltamethrin + heptenophos (16 + 1) (Decisquick) or dimethoate (an organophosphorus insecticide). Resistance to dimethoate depends solely on levels of esterase-based resistance, while resistance to Decisquick depends on kdr and esterase. None of the four insecticides is effective against aphids carrying MACE combined with extreme esterase-based resistance. This knowledge, in association with current monitoring of the mechanisms, will play an important role in making decisions on insecticide use against M persicae in the UK. © 1999 Society of Chemical Industry     

Detection of cyclodiene insecticide resistance-associated mutations by single-stranded conformational polymorphism analysis

Cyclodiene insecticide resistance is associated with replacements of a single amino acid within the putative lining of a δ-aminobutyric acid (GABA)-gated chloride ion channel gene Resistance to dieldrin (Rdl). Only two resistance-associated amino acid replacements have been identified; alanine to serine in Drosophila melanogaster, D. simulans, Aedes aegypti, and Tribolium castaneum and alanine to glycine as a second allele in D. simulans. Here we report that single stranded conformational polymorphism (SSCP) analysis of genomic DNA, amplified by the polymerase chain reaction (PCR) for exon 7 of the Rdl gene, can be used to genotype strains or individuals of all of these insects. This technique also appears simultaneously to distinguish between D. melanogaster and D. simulans, sibling species only reliably identifiable by examination of male genitalia. The relative advantages of this genotyping technique against other PCR-mediated techniques in monitoring for insecticide resistance are discussed.     

The Red Queen in a potato field: integrated pest management versus chemical dependency in Colorado potato beetle control

Originally designed to reconcile insecticide applications with biological control, the concept of integrated pest management (IPM) developed into the systems‐based judicious and coordinated use of multiple control techniques aimed at reducing pest damage to economically tolerable levels. Chemical control, with scheduled treatments, was the starting point for most management systems in the 1950s. Although chemical control is philosophically compatible with IPM practices as a whole, reduction in pesticide use has been historically one of the main goals of IPM practitioners. In the absence of IPM, excessive reliance on pesticides has led to repeated control failures due to the evolution of resistance by pest populations. This creates the need for constant replacement of failed chemicals with new compounds, known as the ‘insecticide treadmill’. In evolutionary biology, a similar phenomenon is known as the Red Queen principle – continuing change is needed for a population to persevere because its competitors undergo constant evolutionary adaptation. The Colorado potato beetle, Leptinotarsa decemlineata (Say), is an insect defoliator of potatoes that is notorious for its ability to develop insecticide resistance. In the present article, a review is given of four case studies from across the United States to demonstrate the importance of using IPM for sustainable management of a highly adaptable insect pest. Excessive reliance on often indiscriminate insecticide applications and inadequate use of alternative control methods, such as crop rotation, appear to expedite evolution of insecticide resistance in its populations. Resistance to IPM would involve synchronized adaptations to multiple unfavorable factors, requiring statistically unlikely genetic changes. Therefore, integrating different techniques is likely to reduce the need for constant replacement of failed chemicals with new ones. © 2014 Society of Chemical Industry     

草地贪夜蛾抗药性现状及化学防治策略

本文就草地贪夜蛾抗药性研究的历史、现状以及如何进行化学防治进行了比较系统的分析。针对国内外草地贪夜蛾发生的现状,从抗药性程度及交互抗性、种群遗传、抗药性机制以及化学防治的关键技术等方面进行了讨论。提出了化学防治要尽量做到药剂品种、时间和空间的配合;掌握好药剂防治的两个窗口期,一是害虫本身敏感的窗口期即从孵化到3龄初,二是孵化后到钻蛀前;分阶段选择适宜药剂类型用于化学防治。除了考虑杀虫剂作用机制类别外,作用方式也要考虑。卵高峰期施用具有触杀活性的药剂配合具有杀卵活性的药剂添加具有渗透功能的助剂,孵化高峰期施用触杀药剂配合胃毒药剂,后期大龄幼虫可以考虑胃毒药剂为主的化学防治策略。