害虫抗药性适合度与内分泌调控研究进展

本文就害虫抗药性适合度与内分泌调控研究进展作了概述。害虫抗药性品系与敏感品系之间在形态、生理反应和生物学特性等方面存在差异 ,通常被认为是抗性基因适应性代价的表现 ,并可能由抗性基因引起的抗性害虫体内生理代谢平衡发生变化产生的。目前就抗性代价的形成机制存在 2种假说 :资源限制假说和干扰代谢平衡假说。害虫抗性和敏感品系的生物适合度差异与其体内激素水平及调控能力变化有关。激素对抗药性相关酶系表达调控的可能机制可分为三类 ,即直接调控、间接调控和共同调控     

温度对朱砂叶螨二种抗药性品系发育和繁殖的影响

朱砂叶螨敏感品系(S)和抗药性三氯杀螨醇品系(Rd)、抗双甲脒品系(Ra)的各螨态历期在一定温度范围内均随温度的增加而缩短,发育速率对温度敏感性的高低依次为Rd>Ra>S。在相同温度条件下,2个抗药性品系均比敏感品系发育更快。3个朱砂叶螨品系的平均每雌总产卵量与温度的关系,可用二次抛物线方程拟合。在相同温度条件下,2个抗药性品系的生殖力均低于敏感品系,这表明抗药性朱砂叶螨具有生殖不利性。     

桔小实蝇抗多杀霉素的生物适合度代价

桔小实蝇Bactrocera dorsalis ( Hendel) 是中国南方地区果蔬重要害虫之一,严重威胁农业生产。多杀霉素是防治桔小实蝇的重要药剂之一,近年来由于在农业生产中大量使用,田间防治效果已有所下降,已出现了抗药性现象。为明确桔小实蝇对多杀霉素的抗药性发展规律,本研究比较了桔小实蝇敏感品系和抗性品系的发育、生殖和存活差异。结果显示：抗多杀霉素桔小实蝇品系具有死亡率增加、部分虫态发育历期延长和繁殖力下降等适合度代价。抗性品系的内禀增长率由0.19下降至0.15,相对适合度降至0.62,表明桔小实蝇对多杀霉素存在较低的抗性风险。     

杀虫双和杀螟丹选育对小菜蛾抗药性的形成及其抗性机制

用杀虫双和杀螟丹在实验室以点滴法处理小菜蛾Plutella xylostella L.四龄幼虫,以连续继代药剂淘汰选育其抗药性。至35代,药剂汰选的小菜蛾对杀虫双和杀螟丹的抗药性较选育前正常品系分别提高了51倍和25倍。其抗药性的形成发展均呈S形,可认为已成为抗性品系。以有机磷、氨基甲酸酯、拟除虫菊酯及有机氮等11种杀虫剂测试抗杀虫双小菜蛾品系和抗杀螟丹小菜蛾品系对常用药剂的敏感度结果表明：对杀虫双、杀螟丹和杀虫环之间有较严重的正交互抗性;对敌敌艮、马拉硫磷和杀螟松有轻微交互抗性产生;对溴氰菊酯、氯氰菊酯、氯菊酯和灭多威、久效威等药剂更加敏感,呈负交互抗性。用聚丙烯酰胺凝胶电泳(PAGE)法测定表明,抗药性产生与特异性酯酶的形成有一定关系。用比色法和酸度法测定,抗性品系的乙酰胆碱酯酶(AchE)活性降低,羧酸酯酶(CarE)活性无差异。加增效剂Pb和SV₁：于四龄幼虫表皮,对抗杀虫双小菜蛾晶系分别有6.28及1.45倍的增效作用;对抗杀螟丹小菜蛾品系分别有4.85及1.39倍的增效作用,可见多功能氧化酶(MFO)为小菜蛾抗杀虫双和抗杀螟丹的重要因子。     

二点叶螨对甲氰菊酯、氧乐果和四螨嗪抗药性的选育、衰退和恢复

以兰州吐鲁沟公园的二点叶螨Tetranychusurticae为敏感品系 ,分别用氧乐果、甲氰菊酯、四螨嗪喷雾处理 15次 ,其抗性水平分别为 38 5、 479 8和 6 7 3倍。将抗性品系分别与敏感品系进行杂交和回交的结果表明 :抗氧乐果品系的显性系数DRS (R♀×S♂ )为 0 470 0、DSR (S♀×R♂ )为 0 4749;抗甲氰菊酯品系的DRS (R♀×S♂ )为 0 5 15 5 ,DSR 为0 5 2 37;抗四螨嗪品系的DRS为 0 3134 ,DSR 为 0 2 46 6。表明二点叶螨对这 3种药剂的抗性均是由单个不完全显性基因所控制。在连续 10个月不接触药剂的情况下 ,3个抗性品系的抗药性都有下降 ,抗氧乐果品系的抗性下降最快 ,只有敏感品系的抗性倍数的 3 6倍 ;抗甲氰菊酯种群的抗性下降较慢 ,为敏感品系的 95 9倍。再经甲氰菊酯、氧乐果、四螨嗪分别连续15次喷雾处理后 ,3个抗性种群的抗性水平又再度回升 ,抗甲氰菊酯品系回升较快 ,抗性为敏感品系的 5 2 3 5倍 ,抗四螨嗪品系次之 ,抗氧乐果的品系抗性恢复最慢。     

桔小实蝇抗性个体流动对抗性个体频率的影响

为了解害虫个体的迁移对害虫群体抗药性发展的影响, 本研究利用实验室内建立的桔小实蝇Bactrocera dorsalis对敌百虫和高效氯氰菊酯两种抗性品系以及相对敏感品系, 设计5%~25%个体的迁移比例, 研究桔小实蝇抗、 感个体流动对原始种群抗性个体频率的影响。结果表明: 桔小实蝇抗性个体迁入敏感种群, 使得敏感种群抗性个体频率增加, 在抗性个体迁移率为25%时, 影响敏感种群敌百虫和高效氯氰菊酯抗性个体频率的变化值分别为20.04%和41.75%; 同样随着敏感个体迁入比例的增加, 抗性种群中抗性个体频率降低程度越大, 在敏感个体迁移率为25%时, 抗性种群敌百虫和高效氯氰菊酯抗性个体频率的变化值分别为56.20%和25.88%。利用抗性个体频率变化值与相应迁移率的比值来表示迁移的相对效率, 在抗性个体迁移率为5%时, 影响敏感种群抗性个体频率变化的相对效率最高; 在敏感个体迁移率分别为5%和10%时, 影响敌百虫抗性种群和高效氯氰菊酯抗性种群抗性个体频率变化的相对效率最高。以抗、 感个体迁移引起的抗性个体频率变化值进行趋势拟合, 发现抗敌百虫与抗高效氯氰菊酯桔小实蝇品系不同迁移比例下的抗性个体频率变化趋势分别符合密度模型(density model)和房屋模型(housing model), 相关系数分别是0.9696和0.9647。研究结果表明通过合理地设计抗、 感迁移比例能有效地延缓桔小实蝇抗性水平的上升, 达到抗性治理的目的。     

果树害虫抗药性及对策

<正> 多年来,农药在害虫防治中起了巨大作用。然而,害虫抗药性乃是化学防治中的一个突出问题。关于害虫抗药性的研究,FAO专门成立了害虫抗药性专家工作组,其主要任务之一就是负责制定农业害虫抗药性监测和测定的标准方法”。就果树害虫而言,1970年发表了桃蚜Myzus persicae (Sulzer)成虫的抗药性测定方     

白纹伊蚊抗高效氯氰菊酯和抗敌百虫品系的生物学特性及种群参数

白纹伊蚊Aedes albopictus Skuse抗高效氯氰菊酯和抗敌百虫品系的某些种群参数及生物学特性与敏感品系的比较有显著差异,两个抗药性品系之间差异也较明显。抗高效氯氰菊酯品系(Rc)的繁殖力强、平均世代历期短,雌性个体比率高。抗敌百虫品系(Rt)有适合度缺陷、平均世代历期长,雌性个体比率低。Rc和Rt的孵化率均低于敏感品系。成虫死亡率由高到低依次为Rt、Rc、S。综合分析得知,高效氯氰菊酯刺激白纹伊蚊生殖、种群增长快。抗敌百虫白纹伊蚊明显地表现出生殖不利性,种群加倍较慢。     

淡色库蚊生物学的实验研究

<正> 淡色库蚊Culex pipiens pallens Coquillett在我国不少地区已形成不同程度的抗药性。为了解它的正常(敏感性)品系和抗药性品系幼虫发育所需要的时间、雌雄性别的比例、成虫在不同温度时的寿命以及其产卵等情况,我们进行了此项实验研究。     

桔小实蝇抗性品系的微卫星DNA分析

用微卫星标记对桔小实蝇 Bactrocera dorsalis(Hendel) 敏感品系和3个抗性品系（敌百虫抗性品系、高效氯氰菊酯抗性品系和阿维菌素抗性品系）进行了基因组筛查。结果表明,抗性品系显示出微卫星DNA多态性,不同的抗性品系已经产生了不同的遗传变异。该结果有利于指导控制桔小实蝇抗药性产生以及抗药性的田间监控,同时在抗药性产生的遗传学机制研究方面有一定的意义。     

Biochemical characterization and purification of esterases from three strains of German cockroach,Blattella germanica (Dictyoptera: Blattellidae)

Three strains of German cockroach, Blattella germanica (L.) showed varying levels of resistance to chlorpyrifos, methyl parathion, propoxur, bendiocarb, and cypermethrin. The general esterase activity was at least twofold higher than susceptible strain. The subcellular distribution studies revealed that the majority of the esterase activity is present in the 100,000g cytosolic fraction. Only a small portion of the activity was membrane bound. Using non-denaturing gel electrophoresis, ten isozymes were identified in German cockroaches. These isozymes were isolated individually from the gels and analyzed for differences in activity. The isozymes E5, E6, and E7 of resistant strains had significantly higher specific activities when compared with the susceptible strain. The purification process using various column chromatography and preparative gel electrophoresis resulted in 9–11% of total esterase recovery. About double the amount of E6 was recovered from the resistant strains when compared with the susceptible strain. Kinetic analyses of E6 did not indicate differences in K_m and V_max values between the resistant and susceptible strains. Also, inhibition of esterase activity by paraoxon, chlorpyrifos, and propoxur did not suggest any structural differences in esterase E6 between strains. The results suggest that the increased production of E6 esterase contributes to insecticide resistance in German cockroaches. The role of E6 may be sequestration of toxic molecules rather than hydrolysis. © 1996 Wiley-Liss, Inc.     

Electrophoretic analysis of esterase isozymes in organophosphate-resistant mosquitoes (Culex pipiens)

Esterase isozymes from individual mosquitoes, Culex pipiens, were analyzed by thin layer agar gel electrophoresis. Changes in esterase isozyme patterns during the developmental stages were studied. In most strains, only one band was found in eggs. During larval development many bands were gradually formed, and the most anodal bands were lost upon pupation. These larva-specific esterases may be controlled by three or more co-dominant alleles. In the adult, many bands were present that were probably controlled by at least seven loci. Est-2 may be controlled by co-dominant alleles; a recessive silent allele was also found.Esterase isozyme patterns of organophosphorus insecticide-resistant and susceptible strains were compared. Resistant strains had very active esterase bands but different patterns were found in malathion and temephos-resistant strains. The strong bands presumably hydrolyse the insecticides to which the strains are resistant, or their PO analogues. Their substrate specificity was studied with various esters.     

Pirimiphos-methyl resistance in two stored product mites, Acarus siro and Acarus farris, as detected by impregnated paper bioassay and esterase activity assays

The response to pirimiphos-methyl, in one strain of Acarus farris and two strains of Acarus siro, was assessed using an impregnated filter paper bioassay and by the selection of adults following exposure to pirimiphos-methyl. It was concluded that one of the strains of A. siro was resistant to pirimiphos-methyl and that a major resistance mechanism was involved. The second strain of A. siro gave a response similar to that of a laboratory strain unexposed to organophosphates and was considered to be susceptible. The A. farris strain responded to selection at the ED50 but not at the ED99, and it was concluded that a minor resistance mechanism is present in this strain. Assays of esterase activity were used to attempt to identify the biochemical mechanisms involved in the resistance detected by the bioassays. The A. farris and susceptible A. siro strains showed similar levels of esterase activity but the esterase activity of the resistant A. siro strain was significantly greater. An increase in esterase activity followed selection of both the A. farris strain and the resistant A. siro strain. An acetylcholinesterase assay showed no significant difference between the susceptible and pirimiphos-methyl selected strains of A. siro. The results suggest that esterases are involved in the resistance to pirimiphos-methyl found in A. siro and A. farris but that in A. siro, at least, other mechanisms may also be present.     

不同种群棉铃虫三龄幼虫酯酶活性频率分布与抗药性的关系

选用室内饲养的棉铃虫Helicoverpa armigera偃师和湖北2个敏感品系、对辛硫磷高抗的PCP20品系、对氰戊菊酯高抗的YG45品系及1999或2000年采自山东阳谷、河北邯郸和河南安阳的田间高抗种群,江苏徐州、湖北武汉的田间中等抗性水平种群和新疆沙湾的田间敏感种群,采用酶标板酶动力学法测定了各品系（种群）的3龄幼虫个体酯酶活性频率分布和平均酯酶活性。结果表明,偃师敏感品系、湖北敏感品系和新疆沙湾田间敏感种群的酯酶活性个体频率分布相似,三个品系（种群）的平均酯酶活性相近,分别为991、1 138、1 055 mOD·min^-1·larva^-1。室内选育的PCP20抗性品系、YG45抗性品系及山东阳谷、河北邯郸 、河南安阳田间高抗种群的高酯酶活性（活性在1 800 mOD·min^-1·larva^-1以上）个体频率明显高于三个敏感品系（种群）,平均酯酶活性在1 510～2 482 mOD·min^-1·larva^-1之间。江苏徐州、湖北武汉的田间中抗水平种群高酯酶活性个体频率及平均酯酶活性都介于敏感和高抗品系（种群）之间,平均酯酶活性为1 258～1.404 mOD·min^-1·larva^-1。棉铃虫各品系（种群）平均酯酶活性与对拟除虫菊酯类杀虫剂抗性个体频率的相关性要比对有机磷类的高,相关系数分别为0.82和0.42。分析各品系（种群）高酯酶活性个体频率与棉铃虫对拟除虫菊酯类、有机磷类杀虫剂抗性个体频率的相关性,得到相似的结果。考虑到酯酶并不是棉铃虫对拟除虫菊酯抗性的主要机理,建议酯酶活性可作为棉铃虫抗药性生化检测的一个参考指标。本文还讨论了酯酶与棉铃虫对拟除虫菊酯类杀虫剂及有机磷类杀虫剂抗性的关系。     

The effects of growth conditions on the esterases of Paramecium aurelia

The esterase isozymes of Paramecium aurelia grown in the presence and absence of bacteria were examined by starch gel electrophoresis. Where present, bacteria make a positive contribution to the esterase zymograms. Different bacteria contribute different esterases. Paramecia growing in axenic medium possess a different esterase which is not produced by the same parmecia growing in the presence of bacteria. This axenic esterase disappears when bacteria are added back to the medium.Supported by a research grant from the Science Research Council.     

杂交水稻及三系在发育过程中的酯酶同工酶比较研究

本文报道了利用聚丙烯酰胺凝胶电泳技术,测定杂交水稻及三系亲本共50个组合的萌动胚、芽,不同发育时期的叶片、根、雄蕊等12个组织或器官的酯酶同工酶的结果。根据所测结果,可把杂交水稻的酯酶同工酶酶谱分为5种类型:互补型、偏父型、偏母型、同型和“杂种”酶谱。强优势组合以互补型酶谱居多,弱优势组合都是同型酶谱,“杂种”酶谱仅见于V优64的幼穗分化期叶片和V优63、汕优63的三叶期叶片中。不同器官的互补酶谱都可作为预测杂种比势,鉴定杂交稻种子的纯度和真实性以及选配新杂交组合的一个手段或依据,但以对萌动胚或幼芽的测定更有实践意义。     

Antioxidative enzymes and isozymes analysis of taro genotypes and their implications in <Emphasis Type="Italic">Phytophthora</Emphasis> blight disease resistance

Assessment of the differential expression of antioxidative enzymes and their isozymes, was done in 30 day-old ex vitro raised plants of three highly resistant (DP-25, Jhankri and Duradim) and one highly susceptible (N-118) genotypes of taro [Colocasia esculenta (L.) Schott]. Antioxidative enzymes were assayed in the ex vitro plants, 7 days after inoculation with the spores (15,000 spores ml⁻¹ water) of Phytophthora colocasiae Raciborski to induce taro leaf blight disease. Uninoculated ex vitro plants in each genotype were used as control. The activity of superoxide dismutase (SOD) and guaiacol peroxidase (GPX) increased under induced blight condition when compared with control. Increase in antioxidative enzymes was more (67–92%) in the resistant genotypes than that (21–29%) of the susceptible genotype. The zymograms of SOD and GPX in the resistant genotypes, with pathogenic infection, showed increased activity for anodal isoform of SOD and increased expression and/or induction of either POX 1 or POX 2 isoforms of GPX. In susceptible genotype, expression of the above isoforms was faint for SOD and nearly absent for GPX under both blight free and induced blight conditions. Induction and/or increased activity of particular isoform of SOD and GPX against infection of Phytophthora colocasiae in the resistant genotypes studied led to the apparent conclusion of linkage of isozyme expression with blight resistance in taro. This might be an important criterion in breeding of taro for Phytophthora leaf blight resistance.     

Plasticity in biochemical characteristics of cotton aphid,Aphis gossypii Glover in response to different host plants and neonicotinoid pesticides

In the present investigation an effort was made to realise the role of feeding host plants on some enzymes’ activities. The results showed that the enzymes’ activities were changed in the aphids feeding on different host plants which assist in detoxification of their host metabolites. It is important when the aphids are exposed to insecticides. The results indicated that the measured enzyme activity has significant changes depending on the host plant. It is shown in this study that there are no significant differences between different host plants on esterase activity (p = 0.446); however, there is a significant difference between GSH activity (p = 0.047) but this relationship is not significant on MFO activity (p = 0.417). Among three strains of Ag-PP, Ag-MO and Ag-FA, strain Ag-PP was the most resistant strain against neonicotinoids, and the resistance mechanism was related to metabolic increase in carboxyl esterase activity. The results showed that strain of Ag-MO was the most susceptible strain against neonicotinoids. The result of this investigation also showed that the general esterases might play an important role in conferring or contributing to neonicotinoid resistance in the cotton aphids.     

Decapitation impacting effect of topically applied chlorpyrifos on acetylcholinesterase and general esterases in susceptible and resistant German cockroaches (Dictyoptera: Blattellidae)

The effect of topically applied chlorpyrifos on acetylcholinesterase and other esterases in heads and decapitated bodies of CSMA and Crawford German cockroaches was examined with spectrophotometric enzyme assay and native polyacrylamide gel electrophoresis. The toxicity of chlorpyrifos was greatly reduced in decapitated CSMA male cockroaches with LD50 value 17.1-fold higher than that of normal CSMA cockroaches. Acetylcholinesterase activity from heads was significantly higher in the Crawford compared with the CSMA strain and did not change until 24 h after chlorpyrifos in vivo treatment in both strains. The p-nitrophenyl butyrate (NPB) esterase activities from both heads and decapitated bodies of the resistant Crawford strain were significantly greater than the susceptible CSMA strain. The p-NPB esterase activity was significantly inhibited by chlorpyrifos in vivo treatment, and total p-NPB esterase activity was significantly reduced in decapitated bodies compared with heads of both strains. Native polyacrylamide gel electrophoresis (PAGE) analysis of extracts solubilized with Triton X-100 from heads and decapitated bodies revealed five major esterase bands and an acetylcholinesterase (AChE) band with a high capability of hydrolyzing alpha-naphthyl butyrate and acetylthiocholine, respectively. In the heads of susceptible CSMA male cockroaches, the activity of mobile isozymes d1 and d2 was completely inhibited at 24 h after chlorpyrifos application, and isozyme e was partially inhibited. In contrast, isozymes c1 and c2 from the decapitated bodies of CSMA cockroaches were mostly affected at 24 h after the topical application of chlorpyrifos. The activities of acetylcholinesterase and esterase isozymes a and b from the decapitated body remained uninhibited in both strains. Inhibition of isozymes d1 and d2 seems to be more important in chlorpyrifos intoxication than acetylcholinesterase.