褐飞虱对甲胺磷、扑虱灵的抗药性现状及发展趋势

本文报道采用点滴法和浸苗法监测江苏地区褐飞虱对甲胺磷、扑虱灵抗药性的现状及发展趋势。从１９９３年到１９９５年褐飞虱对甲胺磷的抗药性指数为４．９～１０．２，属耐药到低抗性水平；对扑虱灵的抗药性指数为２．４～３．１，耐药性明显增加。经连续筛选８代，褐飞虱对甲胺磷和扑虱灵的抗性分别增加５倍和６．５倍，显示这２种药剂大面积连续使用抗药性会继续增加，其中对扑虱灵的抗药性上升速度略快于对甲胺磷。     

水稻品种抗性对虫体酯酶的影响

采用苗期群体鉴定法对供试水稻品种进行了抗性鉴定,在此基础上测定了取食不同抗性水稻品种的褐稻虱(Nilaparvata lugens)羧酸醑酶、酸性及碱性磷酸酯酶活力的变化。结果表明,取食感虫品种的褐稻虱若虫及第2代若虫体内上述三种酶的活力明显高于取食抗虫品种的;取食中抗品种的褐稻虱若虫三种酶的活力介于取食感虫和抗虫品种的若虫之间。这一结果与褐稻虱在不同抗性水平上水稻品种的存活率和发育进度是一致的。表明抗虫品种对褐稻虱具有抗生机制,从而抑制了虫体的正常生长发育。取食不同水稻品种的褐稻虱的羧酸酯酶米氏常数(Km)值无明显差异,但最大反应速度(Vmax)值差异极显著。取食感虫品种的 Vmax 值明显高于取食抗虫品种的。这表明取食不同抗性水稻品种的褐稻虱羧酸酯酶性质相同,差异主要是由于酶量的不同。     

棉铃虫不同龄期幼虫羧酸酯酶、谷胱甘肽转移酶、乙酰胆碱酯酶研究

作者报道了棉铃虫抗性种群(HJ-R)和相对敏感种群(HD-S)幼虫4个发育阶段羧酸酯酶、谷胱甘肽转移酶、乙酰胆碱酯酶的特性。随虫龄的增长,两个种群单头幼虫羧酸酯酶、谷胱甘肽转移酶的酶活力急剧增长;HJ-R种群与HD-S种群谷胱甘肽转移酶酶活力比值逐渐增大。HJ-R与HD-S种群间乙酰胆碱酯酶对杀虫剂的敏感性没有显著差异,但两种群幼虫从4日龄发育到3龄时,乙酰胆碱酯酶发生突变,对杀虫剂的敏感性显著降低。     

褐飞虱对甲胺磷和噻嗪酮代谢抗性的生化分析

作者分析了褐飞虱对噻嗪酮、甲胺磷的代谢抗药性。室内生物测定显示褐飞虱对甲胺磷的抗性与虫体酯酶活力升高有关,而对噻嗪酮的抗性则主要是由于多功能氧化酶活力的提高。褐飞虱的个体酯酶活力测定表明,甲胺磷抗性群体中的酶活力中等以上的个体频率明显增加,达２６．０８％,比对照种群提高１４．６５％;噻嗪酮抗性群体显示相似趋势,但酶活力中等以上的个体频率增幅较小,为２０．７４％,比对照提高９．３１％。说明褐飞虱对这     

褐飞虱对甲胺磷的抗性遗传

室内抗性筛选表明，褐飞虱对甲胺磷抗性的发展呈不规则的“S”形，第5代前抗性上升很慢，第5～15代间迅速上升，第15代后上升再次变慢。利用选育的抗甲胺磷褐飞虱品系和敏感品系，研究了褐飞虱对甲胺磷的抗性遗传形式。结果显示，正交和反交子代的显性度D值分别为0．5130和0．4673，表明抗性主效基因是不完全显性遗传的。利用剂量对数-死亡机率值曲线分析法，初步判断抗性遗传不是单基因控制的；卡方(χ^2)测验进一步证实，抗性遗传可能是两个或两个以上的基因控制。抗性筛选全过程和F5～F13的现实遗传力(h^2)分别为0．2190和0．2457，说明褐飞虱对甲胺磷抗性产生的速率较快，出现高抗性的风险较大。     

桃蚜高效氯氰菊酯抗药性与乙酰胆碱酯酶的关系

于室内对桃蚜进行高效氯氰菊酯抗药性筛选,选育至10代后抗性倍数增长到49.9倍。生化分析表明,抗性品系乙酰胆碱酯酶(AChE)活性均显著高于敏感品系。比较两个品系乙酰胆碱酶活性个体频率分布发现,更多的桃蚜个体向酶活性高的区域分布。酶动力学测定结果显示,抗性桃蚜酯酶对底物的Vmax、Km显著大于敏感品系。     

桃蚜高效氯氰菊酯抗药性与乙胆酯酶的关系

于室内对桃蚜进行高效氯氰菊酯抗药性筛选,选育至10代后抗性倍数增长到49.9倍.生化分析表明,抗性品系乙酰胆碱酯酶(AChE)活性均显著高于敏感品系.比较两个品系乙酰胆碱酶活性个体频率分布发现,更多的桃蚜个体向酶活性高的区域分布.酶动力学测定结果显示,抗性桃蚜酯酶对底物的Vmax、Km显著大于敏感品系.     

嗜卷书虱和嗜虫书虱酯酶性质的比较研究

对嗜卷书虱Liposcelis bostrychophila和嗜虫书虱L. entomophila羧酸酯酶及乙酰胆碱酯酶(AChE)的生物化学性质进行了初步研究。结果表明:嗜虫书虱的羧酸酯酶活性明显高于嗜卷书虱,嗜卷书虱和嗜虫书虱羧酸酯酶对底物α-NA的Km值分别为0.665 7、0.285 7 mmol/L,Vmax值分别为1.672 2和2.463 1 mmol·(mg·30 min)-1。嗜卷书虱和嗜虫书虱乙酰胆碱酯酶的活性差异不显著,嗜虫书虱略高于嗜卷书虱。通过聚丙烯酰胺凝胶电泳,检测出8条嗜卷书虱、7条嗜虫书虱酯酶同工酶谱带,嗜卷书虱的酶谱分布比嗜虫书虱广。     

甲胺磷对不同抗性水平褐飞虱品系种群增长的影响

用点滴法测定了甲胺磷对3个褐飞虱品系的毒力,并用5个剂量(LD5、LD10、LD25、LD50和LD75)处理4龄若虫,存活若虫饲养至羽化后单对继续饲养,观察并记录3个品系的繁殖力。结果表明,甲胺磷具有刺激褐飞虱种群增长的作用,但对不同抗性水平品系的刺激作用有很大差异,对田间种群和抗性品系的刺激作用明显小于敏感品系,其中对抗性品系种群增长的刺激作用尤小。     

甲胺磷对褐飞虱酯酶时序变化的影响

褐飞虱经甲胺磷处理后 ,酯酶活性首先表现为受到抑制 ,然后上升 ,到达最大值后再下降并停留在一个相对对照较高的水平。在 1次处理的基础上 ,第 3天和第 7天再次处理 ,酯酶的变化进程与 1次处理相同 ,但最后的停留水平均高于 1次处理 ,分别为 1次处理的 1.35倍和 1.63倍左右。敏感性测定同样表明再次处理可以导致敏感性进一步下降 ,第 3天和第 7天再次处理 ,敏感性分别比 1次处理下降了0.35 1、0.696倍。     

田间菜蛾绒茧蜂对有机磷敏感性监测及毒理机制分析

对甲胺磷敏感性的田间监测结果显示,绒茧蜂存在着抗性演化,毒力生物测定结果与AChE的K_i值的监测结果呈明显的相关性,每年9月至次年2月期间AChE敏感性最低,8月期间敏感性最高。甲胺磷可显著地抑制绒茧蜂AChE、CarE和GSTs的活性。PB和TPP对AChE的活体抑制率极低,但PB可强烈抑制CarE的活性,而TPP仅在高浓度时对CarE有较显著的抑制作用,PB对甲胺磷有显著的增效作用,而TPP对甲胺磷无增效作用。AChE的K_m、V_(max)及K_i值研究结果表明,田间绒茧蜂对有机磷和氨基甲酸酯的抗性与AChE对杀虫剂的不敏感性有关。由此认为,绒茧蜂对有机磷的抗性主要与其最重要的靶标酶AChE的敏感性改变及多功能氧化酶有关。     

Susceptibilities to methamidophos and enzymatic characteristics in 18 species of pest insects and their natural enemies in crucifer vegetable crops

The susceptibilities to methamidophos as well as the kinetic and inhibitory parameter of acetylcholinesterases (AChE) and the activities of carboxyestsrases (CarE) and glutathione-S-transferases (GST) were studied in 18 species field populations of insects collected in Fuzhou, China during April and May 2000 and 2001. The insect species included five hymenopteran endoparasitoids, one hymenopteran exoparasitoid, one hymenopteran hyperparasitoid, one dipteran predator, four coleopteran predator ladybirds, six herbivorous pest insects of lepidoptera, diptera, homoptera, and coleoptera, respectively. There existed significant correlations between the susceptibility to methamidophos and the k_i values of AChE to methamidophos, dichlorvos, and carbofuran and between the k_i and V_max values of AChE among 18 species of insects. The six herbivorous pests and four ladybirds showed significantly low k_i and V_max values of AChE compared to the seven parasitoids and predator Epistrophe balteate. It was difficult to correlate the susceptibility to methamidophos or the k_i values with the K_m values of AChE, or with the activity of CarE and GST. The activities of CarE and GST varied depending on the different insect species. Significant synergisms of piperonyl butoxide (PB), triphenyl phosphate (TPP), and diethyl maleate (DEM) with methamidophos were observed in 14 pest insects and their natural enemies. Synergisms of PB were found to be the greatest. Reduced k_i values suggested that insensitive AChE might play a critical role in the tolerance to methamidophos in the 18 insect species. The detoxification enzymes, mixed-function oxidase (MFO), CarE, and GST, were believed to be involved in the tolerance to methamidophos. MFO might play the most important role, and CarE or GST might be important in the tolerance in some insect species. Different models of tolerance to methamidophos and enzymatic potential were existed in parasitoids, predators, and herbivores based on the different selection of insecticide pressure (either directly by exposing to the spray in the field, or indirectly by the insecticides penetrated into the body of host insects) as well as different ecological and biological habitats.     

Present status and countermeasures of insecticide resistance in agricultural pests in China

Of the 23 species of agricultural pest known to resist insecticides in China, 4 are cotton pests, 4 rice pests and 5 are pests of brassicae. In the green rice leafhopper, malathion resistance is caused by increased carboxylesterase (CarE) activity, which plays a more important role in the resistance to dimethoate than the mixed-function oxidases (mfos). The in-vitro and in-vivo results are in agreement with studies of synergism of malathion and dimethoate by TPP and EBP. These synergists delay the development of resistance, and EBP when added to malathion has limited the development of resistance to malathion in the green rice leafhopper. In the cotton aphid, resistance to organophosphates involves several factors: acetylcholinesterase (AChE) insensitivity, high CarE activity, slight (× 2) increase in glutathione S-transferases (GSH-ases), mfo activity as well as reduced penetration. In vitro, the I₅₀ of the insensitive AChE is × 14 that of S aphids, and anaphthyl-acetate CarE hydrolysing activity is 70 times greater in R than in S aphids. Insecticide mixtures, alternation or rotation can delay build-up of resistance; resistance to malathion and trichlorfon was delayed in Culex pipiens pallens when the two insecticides were used together. Used singly each insecticide selected for high resistance within 25 generations. Mosaic rotation of dimethoate and fenvalerate delayed the onset of insecticide resistance in Lipaphis erysimi pseudobrassicae.     

小菜蛾和绒茧蜂乙酰胆碱酯酶对甲胺磷和甲基异柳磷敏感性监测

对 ACh E的 K m、Vmax和 Ki值的研究结果表明 ,田间小菜蛾和绒茧蜂对甲胺磷和甲基异柳磷的抗性与 ACh E对杀虫剂的不敏感性有关。田间监测结果表明 ,每年 9月至次年 2月期间 ,由于田间杀虫剂选择压较高 ,小菜蛾 ACh E对有机磷较不敏感 ,而在每年的 6月至 8月期间 ,ACh E敏感性有所升高 ,达到全年的最高点。寄生于同一虫源的绒茧蜂 ACh E敏感性的变化也呈明显的相关性。绒茧蜂 ACh E的敏感性显著高于小菜蛾 ,尚未达到高度不敏感水平。当田间选择压极高时 ,可导致小菜蛾 ACh E呈高度不敏感 ,此时绒茧蜂 ACh E也表现出较高的不敏感性。     

不同防治田内不同发育期拟环纹豹蛛乙酰胆碱酯酶及羧酸酯酶的活性比较

利用紫外分光光度法研究了生物防治田和化学防治田内不同发育期拟环纹豹蛛Pardosa pseudoannulata乙酰胆碱酯酶(AChE)和羧酸酯酶(CarE)的活性特征。结果表明,化防田拟环纹豹蛛各发育时期AChE活性均较生防田中的低,其各发育期AChE的Km值也均明显高于生防田种群。从整个发育期来看,幼体的AChE活性最高,卵囊的最低,说明不同发育阶段AChE活性存在较大差异。而CarE的活性正相反,化防田种群卵囊、幼蛛、雌成蛛和雄成蛛的CarE活性均高于相应生防田种群,且幼蛛的CarE活性高于相应成蛛,生防田种群每一发育阶段CarE的Km值也均高于相应的化防田种群。表明化防田中的拟环纹豹蛛有可能对部分农药产生了一定程度的抗药性。     

Resistance mechanisms and associated mutations in acetylcholinesterase genes in Sitobion avenae (Fabricius)

Wheat aphid, Sitobion avenae (fabricius), is one of the most important wheat pests and has been reported to be resistant to commonly used insecticides in China. To determine the resistance mechanism, the resistant and susceptible strains were developed in laboratory and comparably studied. A bioassay revealed that the resistant strain showed high resistance to pirimicarb (RR: 161.8), moderate reistance to omethoate (32.5) and monocrotophos (33.5), and low resistance to deltamethrin (6.3) and thiodicarb (5.5). A biochemistry analysis showed that both strains had similar glutathione-S-transferase (GST) activity, but the resistant strain had 3.8-fold higher esterase activity, and its AChE was insensitive to this treatment. The I₅₀ increased by 25.8-, 10.7-, and 10.4-folds for pirimicarb, omethoate, and monocrotophos, respectively, demonstrating that GST had not been involved in the resistance of S. avenae. The enhanced esterase contributed to low level resistance to all the insecticides tested, whereas higher resistance to pirimicarb, omethoate, and monocrotophos mainly depended on AChE insensitivity. However, the AChE of the resistant strain was still sensitive to thiodicarb (1.7-fold). Thus, thiodicarb could be used as substitute for control of the resistant S. avenae in this case. Furthermore, the two different AChE genes cloned from different resistant and susceptible individuals were also compared. Two mutations, L436(336)S in Sa.Ace1 and W516(435)R in Sa.Ace2, were found consistently associated with the insensitivity of AChE. They were thought to be the possible resistance mutations, but further work is needed to confirm this hypothesis.     

辛硫磷对枸杞木虱的毒力及对其羧酸酯酶的抑制作用

分别采用浸渍法和药膜法测定了辛硫磷对枸杞木虱不同发育期若虫和成虫的毒力,同时测定了辛硫磷对不同龄期枸杞木虱羧酸酯酶（CarE）活性的抑制作用及酶的动力学常数。结果表明:辛硫磷对枸杞木虱的LC50值随若虫龄期的增大而升高,至成虫期再相对降低,其对5龄若虫的LC50值为0.995 mg/L,是对1龄若虫LC50值（0.047 mg/L）的21.2倍。辛硫磷对不同龄期枸杞木虱羧酸酯酶活性的抑制作用I50值及酶的Km和Vmax值也均随着试虫龄期的增大而升高,至成虫期则I50值稍有降低。研究表明,辛硫磷对枸杞木虱的毒力与对其羧酸酯酶活性的抑制作用间存在密切关系,利用辛硫磷防治枸杞木虱时,以选择龄期较低时用药为宜。     

小菜蛾对溴氰菊酯抗性选育及其机理

本研究用溴氰菊酯在室内以点滴法处理小菜蛾４龄幼虫，连续继代药剂淘汰选育其抗药性，至Ｆ６５代，抗药性提高到１１６３倍，已形成高抗生品系。其抗性的形成发展趋势为前期相对缓慢，中期较快，后期迅速增长。于１９９２、１９９３、１９９４年分别以氯氰菊酯、敌敌畏、杀虫双、灭虫剂有明显 正交互抗性，对其它非菊酯类杀虫虫没有产生交互抗性。用聚丙烯酰胺凝胶电泳地（ＰＡＧＥ）测定表明，小菜蛾对溴氰菊酯抗性的产生可能与非     

江苏省水稻二化螟的抗药性测定及对甲胺磷的抗性机制

采用3龄幼虫点滴法测定了江苏省水稻二化螟对杀虫单和甲胺磷的抗性水平，并就二化螟对甲胺磷产生抗性的机制进行了研究，结果表明，高淳和徐州二化螟种群对杀虫单已达高抗水平，其它地区也表现为中等抗生，在所测定的6个种群中，苏州种群对甲胺磷处于低抗水平，其它种群为中抗种群，澌知体增效作用表明，氧化胡椒基丁醚（PBO）对甲胺磷有明显的增效作用，磷酸三苯酯（TPP）和顺丁烯酸二乙酸（DEM）的增效作用不明显，对离体解毒酶的研究表明，R种群幼虫多功能氧化酶（MFO）的O-脱甲基作用的活力比S幼虫高，R幼虫乙酰胆碱酯酶（AChE)的活力与S幼虫无显著差异。以甲胺磷为抑制剂的R幼虫I50是S幼虫的2.52倍，二化螟对甲胺磷的抗性机理可能与MFO的O-脱甲基作用活力增矿山和AChE的敏感性降低有关。     

寄主植物对桃蚜羧酸酯酶和乙酰胆碱酯酶的诱导作用

在１９９５～１９９６年研究了寄主植物对桃蚜［Ｍｙｚｕｓｐｅｒｓｉｃａｅ（Ｓｕｌｚｅｒ）］羧酸酯酶（ＣａｒＥ）和乙酰胆碱酯酶（ＡＣｈＥ）的诱导作用。在试验的甘蓝、茄子和桃树３种寄主植物中，取食甘蓝的桃蚜种群ＣａｒＥ和ＡＣｈＥ活性最高，取食茄子和桃树的桃蚜种群ＣａｒＥ活性没有明显不同，而ＡＣｈＥ活性取食茄子的桃蚜种群明显高于取食桃树的种群。ＣａｒＥ与底物的亲和力是桃树＞茄子＞甘蓝，而ＡＣｈＥ与底物的亲和力则是甘蓝＞茄子＞桃树。ＡＣｈＥ与毒扁豆碱的双分子速率常数（Ｋｉ）值大小顺序为甘蓝＞桃树＞茄子