N'-硝基缩氨基胍类化合物对蚜虫的室内毒力测定及相关酶活性的影响

Su50[N'-硝基-1-异丙基-2-(4-硝基苯亚甲基)氨基胍]和Su56(N'-硝基-1-烯丙基-2-正丙基氨基胍)是利用活性亚结构拼接原理合成的具有新烟碱类和缩氨基脲类杀虫剂共同结构特征的硝基缩氨基胍类化合物。为了解Su50和Su56对蚜虫的生物活性及作用机理,采用叶片浸渍法测定了其对多种蚜虫的杀虫活性,用棉叶涂抹药液后接入棉蚜Aphis gossypii Glover测定了其对棉蚜的拒食活性,并采用生化方法测定了不同亚致死浓度(LC_(10)、LC_(25))Su56对棉蚜羧酸酯酶(CarE)、乙酰胆碱酯酶(AChE)、谷胱甘肽S-转移酶(GSTs)和多功能氧化酶(MFO)的酶活性的影响。结果表明:Su50和Su56对棉蚜A.gossypii/桃蚜Myzus persicae Sulzer,苹果黄蚜Aphis citricola van der Goot和梅大尾蚜Hyalopterus amygdali Blanchard均具有一定的杀虫活性,但毒力差异不显著,其中Su50对棉蚜、桃蚜、梅大尾蚜和苹果黄蚜的LC_(50)值分别为13.0、24.2、62.6和14.3mg/L,Su56的LC_(50)值则分别为30.3、30.2、101和3.23 mg/L;同时Su50和Su56对供试棉蚜均具有一定的拒食活性。生化试验结果表明,Su56在LC_(10)和LC_(25)浓度下对棉蚜的CarE、GSTs和MFO酶活性无显著影响,但在LC_(10)浓度下对棉蚜的ACNE酶活性却有显著抑制作用。     

寄主植物对二斑叶螨解毒酶活性的诱导作用

[目的]明确不同蔬菜寄主对二斑叶螨解毒酶活性的诱导影响,为叶螨的寄主适应性及叶螨对寄主的抗性机制研究提供基础.[方法]将饲养在菜豆上的二斑叶螨分别转移到黄瓜、甘蓝和茄子上饲养3代后,测定包括菜豆在内的4种不同寄主植物对叶螨体内解毒酶系活性的诱导影响.[结果]取食不同寄主植物的二斑叶螨体内解毒酶系活性差异显著.从菜豆寄主转养到黄瓜、甘蓝和茄子上后,后3个寄主上叶螨种群体内谷胱甘肽S-转移酶(GST)活性显著下降,茄子上种群活性最低,菜豆种群是茄子种群的2.75倍;而后3个寄主种群的乙酰胆碱酯酶(AChE)比活力显著上升,茄子种群最高,是菜豆种群的6.03倍.4种供试寄主植物的二斑叶螨种群中,在黄瓜寄主上羧酸酯酶(CarE)和多功能氧化酶(MFO)活性显著高于其他3种寄主植物,CarE活性的高低顺序为黄瓜＞菜豆和甘蓝＞茄子,而MFO活性为黄瓜＞茄子和菜豆＞甘蓝.[结论]取食不同寄主植物可诱导二斑叶螨体内的解毒酶系活性发生不同程度的变异,这可能是二斑叶螨对寄主具有广泛适应性的生理基础之一.     

不同抗性玉米自交系对朱砂叶螨体内解毒酶活性的影响差异

为了明确不同抗性玉米自交系对朱砂叶螨(Tetranychus cinnabarinus)体内解毒酶活性的影响,测定了在8个玉米自交系上繁殖了3代的朱砂叶螨体内乙酰胆碱酯酶(AChE)、多功能氧化酶(MFO)、谷胱甘肽硫转移酶(GST)和羧酸酯酶(CarE)的活性。结果表明,8个不同抗虫性玉米自交系上的朱砂叶螨体内4种解毒酶活性存在一定差异。朱砂叶螨体内AChE和MFO以取食高抗玉米自交系HL6044时为最高,分别为(143.5±5.52)nmol/min·mg、(47.09±2.11)pmol/30min·mg;CarE比活力为(62.00±1.89)ΔOD/min·mg,显著高于取食其余7个自交系(P0.05);而GST比活力以取食中抗性玉米自交系HL8028时最高,为(42.64±1.53)ΔOD/min·mg,其次是高抗自交系HL6044,2者均显著高于其余6个自交系(P0.05)。     

苹果绵蚜田间种群的抗性监测

为了掌握果园重要害虫苹果绵蚜Eriosoma lanigerum的抗药性水平及动态变化,于2012–2015年间利用浸叶法监测了中国新疆察布查尔、山东济南、陕西乾县、河北昌黎和江苏丰县苹果绵蚜对常用药剂吡虫啉、毒死蜱和高效氯氟氰菊酯的敏感性,同时测定了不同苹果绵蚜种群的酯酶和谷胱甘肽S-转移酶活力。以2012年察布查尔苹果绵蚜为相对敏感品系,结果发现:所有种群在不同年度均对吡虫啉产生了低到高水平抗性（最大抗性倍数 >2000倍）;对于高效氯氟氰菊酯,昌黎、乾县及察布查尔2014种群均为敏感,济南、丰县和察布查尔2015种群均产生了中到高水平抗性;对于毒死蜱,乾县种群保持敏感,察布查尔、昌黎、丰县和济南种群产生了低到高水平抗性。酶活力测定结果显示:谷胱甘肽S-转移酶活力与苹果绵蚜对吡虫啉和高效氯氟氰菊酯的抗性水平呈正相关性,而酯酶活力与苹果绵蚜抗药性水平无明显相关性。本研究表明,新疆察布查尔、山东济南、陕西乾县、河北昌黎和江苏丰县的苹果绵蚜对吡虫啉、毒死蜱和高效氯氟氰菊酯的抗性水平均呈逐年提高趋势,其中对吡虫啉的抗性程度最严重,防治中应注意合理轮换用药。     

6%吡·丁硫微乳剂防治苹果蚜虫田间药效试验

在山西省危害苹果树的蚜虫主要是绣线菊蚜(Aphis citricola)又名苹果蚜、苹果黄蚜,此虫以成虫和若虫群集危害苹果幼芽、幼枝顶端及幼叶背面,严重时被害叶的叶尖向背面横卷或向下弯曲,导致新梢生长受阻,削弱树势。为了寻求有效的防治苹果蚜虫的药剂,2003-2004年在山西太原进行了6%吡·丁硫微乳剂防治苹果蚜虫田间药效试验,收到了良好的防治效果。1材料与方法1.1供试农药6%吡·丁硫微乳剂,青岛碱业股份有限公司双收农药分公司产品;20%丁硫克百威乳油(对照农药),湖南海利化工股份有限公司产品(2003年)或山东盛邦鲁南农药有限公司产品(2004年);…     

甲氨基阿维菌素苯甲酸盐亚致死剂量对不同敏感性棉铃虫解毒酶活性的影响

室内用甲维盐对棉铃虫选育20代,获得敏感性降低2.32倍的汰选种群(RP20)。分别采用甲维盐对同源对照种群(CP)和RP20的亚致死浓度LC25对应处理两种群的3龄初幼虫48 h,测定试虫在3~6龄不同龄期内羧酸酯酶(CarE)和谷胱甘肽S 转移酶(GST)活性,分析酶活性变化动态。结果表明,(1)两种群(CP和RP20)均随龄期的增长,CarE比活力显著增大,而GST比活力先降低后趋于稳定。(2)CK处理下,RP20与CP相比,CarE比活力在3~6龄均增大且4~6龄差异显著,而GST比活力变化较小,仅3龄和6龄显著增大。(3)与CK处理相比,甲维盐LC25处理,两种群各龄期CarE比活力均增大,且CP试虫在4龄、6龄GST比活力显著增大,在3龄、5龄无显著差异,而RP20的3龄幼虫体内GST比活力被显著抑制,在4~6龄GST比活力无明显变化。初步判断,棉铃虫对甲维盐敏感性降低与CarE比活力增大有关;甲维盐亚致死剂量对棉铃虫CarE活性具有一定的诱导作用。     

太原市不同地区苹果黄蚜抗药性水平监测

本文用FAO推荐的微量点滴法对太原市两地区的苹果黄蚜的抗药性变化进行了监测,结果发现在2010—2013的四年中,不同地区苹果黄蚜对不同药剂的敏感性变化不同。其中清徐地区苹果黄蚜对毒死蜱、乙酰甲胺磷、甲基阿维菌素、氧化乐果、马拉硫磷的敏感性有降低的趋势,其中对氧化乐果的抗性增长最大,为原来的138.94倍,对吡虫啉、啶虫脒、高效氯氰菊酯的敏感性有升高的趋势,其中对啶虫脒的敏感性升高最明显,敏感性升高为原来的27.68倍;阳曲地区黄蚜对吡虫啉、乙酰甲胺磷、甲基阿维菌素、氧化乐果、马拉硫磷的敏感性有降低的趋势,其中对马拉硫磷的抗性增长最快,为原来的151.02倍,对毒死蜱、啶虫脒、高效氯氰菊酯的敏感性有升高的趋势,其中对高效氯氰菊酯的敏感性升高最明显,敏感性升高为原来的1.735 2倍;清徐地区黄蚜对各种药剂的敏感性增长顺序为:氧化乐果乙酰甲胺磷毒死蜱马拉硫磷甲基阿维菌素吡虫啉高效氯氰菊酯啶虫脒,阳曲地区黄蚜对各种药剂的敏感性增长顺序为:马拉硫磷乙酰甲胺磷氧化乐果甲基阿维菌素吡虫啉毒死蜱啶虫脒高效氯氰菊酯。     

土耳其斯坦叶螨对杀螨剂的抗性选育及解毒酶活力变化

为探索土耳其斯坦叶螨的抗药性及其生化机理,在室内对敏感系土耳其斯坦叶螨分别用螺螨酯、甲氰菊酯和阿维菌素逐代处理,选育出抗性种群。结果表明,选育至15代,土耳其斯坦叶螨对螺螨酯、甲氰菊酯和阿维菌素的抗性指数分别达到268.63、37.98和112.68倍。分别测定敏感品系(SS)、抗螺螨酯(RS)、抗甲氰菊酯(RF)、抗阿维菌素(RA)品系的解毒酶活性显示,3种不同抗性品系相对SS品系的羧酸酯酶(CarE)、谷胱甘肽S-转移酶(GSTs)和多功能氧化酶(MFO)的比活力均有不同程度的提高,差异均达到显著水平(P0.05)。其中,RF品系的MFO比活力上升最快,是SS品系的12.7倍;RA品系的MFO比活力次之,是SS品系的5.76倍;RS品系的3种解毒酶比活力均增长较慢,其中CarE比活力上升最慢,是SS品系的1.31倍。由此表明,CarE、GSTs、MFO的活性增大可促进土耳其斯坦叶螨对3种杀虫剂的抗性形成;螺螨酯的抗性增强可能与CarE关系甚微;MFO活性的增加可能与甲氰菊酯抗性升高密切相关;GSTs、MFO的活性升高可能是土耳其斯坦叶螨对阿维菌素产生抗性的主要原因。     

联苯肼酯亚致死质量浓度对二斑叶螨Tetranychus urticae解毒酶系的影响

为明确联苯肼酯对二斑叶螨Tetranychus urticae体内解毒酶系的影响,采用叶片浸渍法测定联苯肼酯对二斑叶螨的亚致死质量浓度LC₁₀和LC₃₀,用这2种浓度分别处理二斑叶螨,并测定60 h内二斑叶螨体内谷胱甘肽S-转移酶（glutathione S-transferase,GST）、羧酸酯酶（carboxylesterase,CarE）和多功能氧化酶（multifunctional oxidase,MFO）的比活力,以及米氏常数K_m及最大反应速率v_max的变化。结果显示,经LC₁₀和LC₃₀剂量处理二斑叶螨后,其体内CarE、GST和MFO的活性在6~60 h均受到明显诱导。其中,GST和MFO的比活力在12 h最大,分别为对照的1.25倍、1.60倍和1.63倍、1.84倍,CarE的比活力在48 h最大,为对照的1.27倍和1.37倍;与对照组相比,CarE的K_m显著减小,v_max显著升高,而GST和MFO的K_m显著增大,v_max显著减小。表明3种酶均参与了二斑叶螨对联苯肼酯的解毒代谢,其中CarE与底物的亲和力最大,反应速率最快,其对联苯肼酯的解毒代谢起主导作用。     

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。     

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.     

Variability in resistance-related enzyme activities in field populations of the tarnished plant bug, Lygus lineolaris

Widespread use of Bt crops for control of lepidopterous pests has reduced insecticide use and provided the tarnished plant bug the opportunity to become a serious pest on mid-South cotton. Organophosphate insecticides have predominantly been used against plant bugs in recent years due to the reduced efficacy of other insecticides. In this study, a biochemical approach was developed to survey enzymatic levels associated with organophosphate resistance levels in field populations of the tarnished plant bug. Forty-three populations were collected from the delta areas of Arkansas, Louisiana, and Mississippi. Three esterase substrates and one substrate each of glutathione S-transferase (GST) and acetylcholinesterase (AChE) were used to determine corresponding detoxification enzyme activities in different populations. Compared to a laboratory susceptible colony, increases up to 5.29-fold for esterase, 1.96-fold for GST, and 1.97-fold for AChE activities were detected in the field populations. In addition to the survey of enzyme activities among the populations, we also examined the susceptibility of major detoxification enzymes to several inhibitors which could be used in formulations to synergize insecticide toxicity against the target pests. As much as 52-76% of esterase, 72-98% of GST, and 93% of AChE activities were inhibited in vitro. Revealing variable esterase and GST activities among field populations may lead to a better understanding of resistance mechanisms in the tarnished plant bug. This study also reports effective suppression of detoxification enzymes which may be useful in future insecticide resistance management program for the tarnished plant bug and other Heteropteran pests on Bt crops.     

Assessment of insecticide resistance in eggs and neonate larvae of Cydia pomonella (Lepidoptera: Tortricidae)

Spanish Cydia pomonella (L.) field populations have developed resistance to several insecticide groups. Diagnostic concentrations were established as the LC₉₀ calculated on a susceptible strain (S_Spain) for five and seven insecticides and tested on eggs and neonate larvae field populations, respectively. The three most relevant enzymatic detoxification systems (mixed-function oxidases (MFO), glutathione S-tranferases (GST) and esterases (EST)) were studied for neonate larvae.In eggs, 96% of the field populations showed a significantly lower efficacy when compared with the susceptible strain (S_Spain) and the most effective insecticides were fenoxycarb and thiacloprid. In neonate larvae, a significant loss of susceptibility to the insecticides was detected. Flufenoxuron, azinphos-methyl and phosmet showed the lowest efficacy, while lambda-cyhalothrin, alpha-cypermethrin and chlorpyrifos-ethyl showed the highest. Biochemical assays showed that the most important enzymatic system involved in insecticide detoxification was MFO, with highest enzymatic activity ratios (5.1-16.6 for neonates from nine field populations). An enhanced GST and EST activities was detected in one field population, with enzymatic activity ratios of threefold and fivefold for GST and EST, respectively, when compared with the susceptible strain. The insecticide bioassays showed that the LC₉₀ used were effective as diagnostic concentrations. Measures of MFO activity alongside bioassays with insecticide diagnostic concentrations could be used as tools for monitoring insecticide resistance in neonate larvae of C. pomonella.     

重庆潼南朱砂叶螨田间抗性监测及其抗性机制分析

本研究通过生物测定检测了重庆潼南田间朱砂叶螨种群对5种杀虫杀螨剂的抗药性,并测定了主要代谢酶活性及抗性相关基因的表达量。抗性监测结果显示,朱砂叶螨对联苯肼酯和哒螨灵表现为中等水平抗性,对阿维菌素、丁氟螨酯和甲氰菊酯为低水平抗性。代谢酶活性检测结果显示:田间种群的P450和GST活性明显提高,且多个相关基因的表达也显著上调,而CarE的活性没有显著变化。综上所述,重庆潼南田间朱砂叶螨种群已对联苯肼酯、哒螨灵、阿维菌素、丁氟螨酯和甲氰菊酯产生了不同程度的抗药性,P450和GST活性提高和有关基因过表达是抗性产生的主要原因。     

Insecticide toxicity and synergism by enzyme inhibitors in 18 species of pest insect and natural enemies in crucifer vegetable crops

The toxicities of three enzyme inhibitors and their synergistic effects on four insecticides were studied by using the dry film method on field populations of 18 species of insects collected in Jianxin and Shanjie, China, from 2003 to 2005. Meanwhile, the inhibitory effects of these enzyme inhibitors on the activities of acetylcholinesterases (AChE), carboxyesterases (CarE) and glutathione-S-transferases (GST), in vivo, were also studied. In general, triphenyl phosphate (TPP) and diethyl maleate (DEM) showed low toxicities to six herbivorous pest insects, four ladybirds and eight parasitoids. Piperonyl butoxide (PB) exhibited low toxicities to the herbivorous pest insects and ladybirds, but high toxicities to the eight parasitoids. The tolerance to the insecticides in 11 pest insects and natural enemies was mainly associated with the tolerance to PB. PB showed the highest synergism on methamidophos, fenvalerate, fipronil and avermectin in nine species of pest insects and natural enemies. In general, TPP and DEM showed significant synergisms to these four insecticides in four parasitoid species. However, in contrast to their effects on the parasitoids, the synergistic effects of TPP and DEM on the four insecticides by TPP and DEM against four pest insects and one ladybird varied depending on the insect species and enzyme inhibitor. Activity of AChE, CarE or GST could be strongly inhibited, in vivo, by PB, TPP or DEM, depending on the insect species and enzyme inhibitors. From the results obtained in this study, mixed-function oxidase (MFO) was thought to play the most critical role in insect tolerances to the tested insecticides in the field. Low competition existed in the evolution of insecticide resistance in the field populations of parasitoids, as compared with herbivorous pest insects and ladybirds. Possible causes of the high synergistic effects of PB on the four classes of insecticides, based on multiattack on the activity of CarE, GST or AChE in the insect species, are also discussed.     

增效剂对菜蚜茧蜂杀虫剂敏感性的影响

以田间菜蚜茧蜂为试虫,采用药膜法研究了增效剂胡椒基丁醚(PB)、磷酸三苯酯(TPP)和马来酸二乙酯(DEM)对6种杀虫剂的增效作用。结果表明,三种增效剂对6种杀虫剂均有显著增效作用,其大小依次为PB>DEM>TPP。PB对甲胺磷、阿维菌素、氟虫腈、氰戊菊酯、氯氰菊酯和吡虫啉的增效比达5.0～9.6倍。TPP和DEM对甲胺磷、DEM对氟虫腈的增效比达2.6～3.0倍,但TPP和DEM对阿维菌素、氰戊菊酯和氯氰菊酯、TPP对氟虫腈的增效比均在1.9倍以下。PB、TPP和DEM对吡虫啉的增效比分别高达9.6、6.8和8.2倍。体内抑制试验结果显示,PB、TPP和DEM对菜蚜茧蜂AChE活性无明显抑制作用,而PB和TPP对羧酸酯酶(CarE)、DEM对谷胱甘肽S转移酶(CST)活性有显著抑制作用。由此认为,菜蚜茧蜂对所用的6种杀虫剂的耐药性与多功能氧化酶(MFO)、CarE和GST的解毒作用有关。     

杀虫增效剂--酶抑制剂对蝶蛹金小蜂和颈双缘姬蜂的影响

采用药膜法测定了3种酶抑制剂对蝶蛹金小蜂和颈双缘姬蜂杀虫剂敏感性的影响。结果表明，胡椒基丁醚、磷酸三苯酯和马来酸二乙酯显著增大了蝶蛹金小蜂和颈双缘姬蜂对甲胺磷、氰戊菊酯、氯氰菊酯、氟虫腈和阿维菌素的敏感性，胡椒基丁醚的作用显著高于后两者，而后两者之间较为接近。2种寄生蜂对氰戊菊酯和氯氰菊酯的击倒作用具明显的恢复能力，而胡椒基丁醚、磷酸三苯酯和马来酸二乙酯可显著抑制这种击倒恢复能力。上述结果表明，2种寄生蜂对所测试杀虫剂的耐药性与多功能氧化酶、羧酸酯酶和谷胱甘肽-S-转移酶的解毒作用有关，其中多功能氧化酶的作用可能是最重要的。     

槲皮素对棉铃虫体内一些解毒酶系和靶标酶的诱导作用

用培养基混药法研究了槲皮素对棉铃虫羧酸酯酶、谷胱甘肽转移酶的诱导作用和对乙酰胆碱酯酶敏感度的影响。槲皮素诱导种群对甲基对硫磷的敏感度明显降低 ,对灭多威和溴氰菊酯的敏感度没有明显变化。诱导种群羧酸酯酶的活性提高 2～ 3倍 ,谷胱甘肽转移酶活性提高近 15倍 ,而乙酰胆碱酯酶对对氧磷的敏感度却提高了 3.5倍。说明槲皮素诱导种群对甲基对硫磷的敏感度降低是由于解毒酶系和乙酰胆碱酯酶综合作用的结果。