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。     

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。     

氰戊菊酯残留对菜蛾绒茧蜂成虫的致死和亚致死效应

室内测定了氰戊菊酯残留对菜蛾绒茧蜂成虫的致死和亚致死效应。结果发现,氰戊菊酯对菜蛾绒茧蜂成虫的毒力比对小菜蛾2龄幼虫的高10倍以上,因此可以肯定田间施用氰戊菊酯后,可以直接(喷雾)或间接(残留)杀死部分菜蛾绒茧蜂成虫。小菜蛾2龄幼虫LC_(50)剂量的氰戊菊酯残留显著降低了雌蜂的产卵量和供试小菜蛾幼虫的被寄生率,在氰戊菊酯残留存在的情况下,雌蜂的死亡率高于雄蜂,蜂子代存活率及性比显著低于对照。氰戊菊酯残留对菜蛾绒茧蜂的搜索行为也有显著的影响,雌蜂在叶片上停留的时间及用于搜索的时间均高于对照,表明残留对菜蛾绒茧蜂雌蜂的搜寻有刺激作用,但产卵寄生的发生次数与对照间无显著差异。文中对产生上述结果的原因进行了讨论。     

温度对几种害虫和寄生蜂乙酰胆碱酯酶对杀虫剂敏感性的影响

田间菜蛾绒茧蜂Cotesia plutellae、颈双缘姬蜂Diadromus collaris和菜蚜茧蜂Diaeretiella rapae对敌敌畏的敏感度及AChE的Ki值远高于其寄主害虫小菜蛾Plutella xylostella和菜缢管蚜Lipaphis erysimi。与对照(25℃)相比,高反应温度(37℃)可显著增高敌敌畏对所测试的5种昆虫AChE的抑制率,而低反应温度(5℃)对5种昆虫AChE的抑制率影响不显著。研究结果还表明,高温对3种寄生蜂AChE的影响明显高于对2种寄主害虫AChE的影响。     

寄主抗药性对菜蛾绒茧蜂寄生能力的影响

考察了在使用氰戊菊酯的条件下 ,小菜蛾抗药性对菜蛾绒茧蜂寄生能力的影响。在杀虫剂作用下 ,用抗性寄主饲养的菜蛾绒茧蜂RRC品系对抗性寄主RRP品系幼虫的寄生率达89% ,寄主幼虫死亡率仅 5% ,比RRC对小菜蛾敏感品系SSP幼虫寄生时、或用SSP饲养的菜蛾绒茧蜂SSC品系对SSP寄生时的寄生率显著要高 ,寄主死亡率显著要低。表明杀虫剂存在时 ,寄主抗药性使其具较高存活率而给寄生蜂提供更多的寄主 ,有利于寄生蜂的繁衍。方差分析结果显示 ,小菜蛾品系及蛾×蜂互作对供试小菜蛾幼虫的被寄生率及死亡率均有显著影响 ,表明小菜蛾抗性改变了其对菜蛾绒茧蜂的适合性     

菜田常用杀虫剂对小菜蛾及菜蛾绒茧蜂的选择毒杀作用

在室内测定了９种菜田常用杀虫剂对小菜蛾幼虫及其主要天敌菜蛾绒茧蜂成虫和茧的毒性。结果表明,应用厂家推荐田间防治小菜蛾的使用浓度,对小菜蛾的杀虫效果锐劲特、Ｂｔ、虫螨光可达１００％,生物复合病毒杀虫剂、宝路和抑太保可达９５％以上,杀虫双达８０％,而氰戊菊酯和万灵对小菜蛾的杀虫效果较差,供试幼虫最终化蛹率分别达７０．０％和７８．３％。让菜蛾绒茧蜂成虫接触厂家推荐浓度的药膜,接触杀虫双２小时后的死亡率即达８０％以上;１２小时后,接触锐劲特的死亡率即达１００％;２４小时后,接触万灵和杀虫双的累计死亡率达９０％以上,接触宝路和氰戊菊酯的约５０％,而接触抑太保、Ｂｔ、虫螨光和生物复合病毒杀虫剂的累计死亡率均在２０％以下,和对照无显著差异。作者认为在菜蛾绒茧蜂等天敌较为丰富的季节,虫螨光、Ｂｔ、生物复合病毒杀虫剂、宝路和抑太保应属防治小菜蛾优先选用的杀虫剂,锐劲特则主要应在小菜蛾大发生、而天敌又很少时应用。     

多杀菌素对菜蛾绒茧蜂的致死和亚致死效应

以对多杀菌素高抗的小菜蛾纯合子品系作寄主饲养菜蛾绒茧蜂,用多杀菌素田间推荐使用浓度(25 mg/L)分别处理处于卵期、幼虫期和蛹期的绒茧蜂,其中卵期和幼虫期通过处理寄主幼虫间接处理,并测定该药剂对成虫的毒力,系统观察该杀虫剂对绒茧蜂各虫期的致死和亚致死作用。与对照相比,绒茧蜂卵期或幼虫期其寄主受药后,可导致蜂幼虫死于寄主体内或啮出寄主的过程中,使结茧化蛹率下降43% ~56%,成虫羽化率下降19% ~33%,成蜂雌性比下降11% ~25%,而成虫大小、寿命和寄生力未受显著影响;在蛹期受药使成虫羽化率下降10%,成蜂寿命缩短,但对成虫寄生力无显著影响;对成虫24 h的LC99为4.31 mg/L。结果表明,多杀菌素可进入寄主作用于其体内的蜂卵和幼虫,或直接作用于蜂蛹和成虫,从而对绒茧蜂各个虫态产生显著的致死和亚致死效应,尤以对成虫高毒。     

十字花科蔬菜—小菜蛾—寄生性天敌三营养级系统研究：不同条件下小菜蛾…

本文根据在深圳、广州、东莞三地对不同蔬菜、不同农事措施、杀虫剂不同施用水平菜地小菜蛾自然种群幼虫被寄生率调查结果，分析十字花科蔬菜－小菜蛾－菜蛾绒茧蜂三种群系统中，蔬菜品种、农事操作、杀虫剂对菜蛾绒茧蜂寄生行为的影响。     

引起小菜蛾盘绒茧蜂雌雄差异反应的气味筛选及行为学检测

小菜蛾盘绒茧蜂Cotesia vestalis是小菜蛾Plutella xylostella幼虫期重要的单性内寄生蜂,在小菜蛾的生物防治上具有广阔的应用前景。寄主植物受到小菜蛾为害后释放的挥发物的种类和释放量会变化,这在小菜蛾盘绒茧蜂搜寻小菜蛾过程中也起到了非常重要的作用。然而,这些寄主植物挥发物对小菜蛾盘绒茧蜂的吸引作用是否存在雌雄差异并不明确。本研究选取了32种十字花科蔬菜受小菜蛾为害后发生变化的气味以及小菜蛾幼虫粪便气味和小菜蛾的性信息素,通过EAG测定了小菜蛾盘绒茧蜂对各种气味物质不同浓度(100、10、1μg/μL)的反应。结果显示,小菜蛾盘绒茧蜂对(E)-4,8-二甲基-1,3,7-壬三烯(DMNT)、苯乙腈、(R)-(+)-柠檬烯、正庚醛4种气味分别在100、100、1μg/μL和1μg/μL的浓度下存在显著雌雄差异。进一步通过"Y"形嗅觉仪对这4种气味进行行为学检验,其中DMNT、(R)-(+)-柠檬烯和正庚醛分别在浓度为10、10μg/μL和0.1μg/μL时可以引起显著的雌雄行为差异。     

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。     

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.     

Insecticide susceptibility of surviving Cotesia plutellae (Hym: Braconidae) and Diaeretiella rapae (M'Intosh) (Hym: Aphidiidae) as affected by sublethal insecticide dosages on host insects

The effects of sublethal dosages of insecticides applied to Plutella xylostella L. (Lepidoptera: Yponomeutidae) and Lipaphis erysimi Kaltenbach (Homoptera: Aphidiidae) on the insecticide susceptibility of the surviving endoparasitoids, Cotesia plutellae Kurdjumov (Hymenoptera: Braconidae) and Diaeretiella rapae (M'Intosh) (Hymenoptera: Aphidiidae), were studied in Shangjie, Minhou, China. The susceptibility to methamidophos and the sensitivity of acetylcholinesterase (AChE) to methamidophos and dichlorvos in the adults of host insects were substantially lower than those in the two parasitoids. The host insects were treated with sublethal dosages of methamidophos in P. xylostella and of methamidophos and avermectin in L. erysimi. The cocoon formation in the two parasitoids decreased significantly, from 35.0% (control) to 13.0% (with methamidophos treatment) for C. plutellae; from 20.6% (control) to 9.0% (with methamidophos treatment) and from 24.3% (control) to 16.7% (with avermectin treatment) for D. rapae. The susceptibility to methamidophos of the resultant emerging adults of the two parasitoids was found to be significantly lower than that of the control when the parasitoids were left in contact with the same dosages of methamidophos. The average AChE activity inhibition by methamidophos and dichlorvos in 34-60 adults of the two parasitoids that emerged from the treatments (15.1% and 31.8% respectively for C. plutellae, and 21.1% and 26.9% for D. rapae) was also significantly lower than those of the controls (55.4% and 48.3% respectively for C. plutellae, and 42.9% and 51.7% for D. rapae). The bimolecular rate constant (k(i)) values of AChE to methamidophos and dichlorvos in the adults of parasitoids without the insecticide treatment were 1.78 and 1.56 times as high as those that emerged from the host insects treated with methamidophos for C. plutellae, and 1.91 and 1.66 times as high as those in the case of D. rapae. It is suggested that there is a difference in AChE sensitivity to insecticides between the resultant emerging parasitoids with and without insecticide pretreatment. Furthermore, the introduction of the insecticides to the host insects could be an important factor in the insecticide resistance development of the endoparasitoids. The natural selectivity would favour the parasitoids that had developed an insensitivity to the insecticide(s).     

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。     

Evaluation of selective toxicity of five pesticides against Plutella xylostella (Lep: Plutellidae) and their side-effects against Cotesia plutellae (Hym: Braconidae) and Oomyzus sokolowskii (Hym: Eulophidae)

The toxicities of five pesticides commonly used in vegetable fields to the larvae of the diamondback moth, Plutella xylostella (L) and its two major parasitoids, Cotesia plutellae (Kurdjumov) and Oomyzus sokolowskii (Kurdjumov), were evaluated in the laboratory using several bioassays. When tested at the rates recommended for field application by the manufacturer, fipronil, avermectin and chlorfluazuron produced over 95% mortality on third-instar larvae of P xylostella, but fenvalerate and methomyl did not produce any substantial mortality to such larvae, and over 70% of P xylostella larvae could still pupate normally after feeding on cabbage leaf treated with the two pesticides. In a residual contact assay with adult parasitoids, fipronil and methomyl caused over 93% mortality within 24 h; fenvalerate killed approximately 55% of the adults within 24 h; avermectin and chlorfluazuron showed differential toxicity to the two parasitoids. Avermectin was extremely harmful to O sokolowskii but slightly toxic to C plutellae, while chlorfluazuron was more toxic to C plutellae than to O sokolowskii. In the residue assay with cocoons of C plutellae and pupae of O sokolowskii developed within P xylostella pupae, treatments with pesticides did not reduce adult emergence. However, the mortality of C plutellae adults emerged from the fipronil-treated cocoons increased greatly within 24 h post-emergence; adults from the fenvalerate-treated cocoons parasitized less host larvae. Treatments with fenvalerate and methomyl significantly increased the mortality of O sokolowskii adults within 24 h post-emergence. Recommendations of choosing pesticides for the control of P xylostella are given.     

杀虫药剂混用对棉铃虫乙酰胆碱酯酶和羧酸酯酶的联合抑制作用研究

选用辛硫磷、对硫磷、敌敌畏和氧乐果4种有机磷类杀虫剂以及灭多威和甲萘威两种氨基甲酸酯类杀虫剂分别以1∶1、1∶3和3∶1的比例混用,以棉铃虫Helicoverpa armigera为试虫,分别测得单剂和混剂对其乙酰胆碱酯酶(AChE)和羧酸酯酶(CarE)的抑制中浓度(I50),以联合抑制系数作为增效作用的参考指标进行了比较。研究发现,两种药剂不同配比的混剂对同种酯酶的联合抑制作用也往往不同。对AChE和CarE同时具有抑制作用的只有其中的对硫磷＋甲萘威(1∶3)、辛硫磷+对硫磷(1∶3)和灭多威+氧乐果(1∶3),而甲萘威+敌敌畏(1∶3)、甲萘威＋氧乐果(1∶1)、辛硫磷+甲萘威(1∶1)、对硫磷+氧乐果(1∶1)、对硫磷+氧乐果(1∶3)、敌敌畏+氧乐果(1∶3)、灭多威+对硫磷(1∶1) 7种混剂对AChE和CarE都具有拮抗作用。     

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.