棉铃虫抗辛硫磷品系的代谢抗性机理

通过重复回交和药剂选择,将棉铃虫Phoxim-R抗性品系对辛硫磷的抗性导入到BK77敏感品系中,得到棉铃虫BK77-R抗性品系,BK77-R和BK77为一对近等基因系。BK77-R抗性品系对辛硫磷的抗性达155倍,对溴氰菊酯有高水平交互抗性(抗性倍数248倍),对灭多威和硫丹有中等水平交互抗性, 分别为31倍和11倍,对丙溴磷有低水平交互抗性(4倍)。在BK77-R抗性品系中,脱叶磷(DEF,酯酶抑制剂)对辛硫磷、灭多威和硫丹具有增效作用,增效倍数分别为7倍、2倍 和1.9倍;增效醚(PBO,氧化酶抑制剂)对溴氰菊酯、灭多威和辛硫磷的增效倍数分别为21倍、2.2倍和1.7倍。与BK77敏感品系相比,BK77-R抗性品系的酯酶和多功能氧化酶活性均显著提高,而谷胱甘肽S-转移酶活性没有明显变化。上述结果表明,酯酶解毒代谢在棉铃虫BK77-R品系对辛硫磷的抗性中起重要作用,酯酶和多功能氧化酶解毒作用增强是该抗性品系对不同类型药剂产生交互抗性的重要原因。     

西花蓟马抗甲氨基阿维菌素苯甲酸盐种群的交互抗性与生化抗性机制

为了解西花蓟马Frankliniella occidentalis(Pergande)对甲氨基阿维菌素苯甲酸盐的抗性风险,采用生物和生化测定方法研究了西花蓟马甲维盐抗性种群与其它杀虫剂的交互抗性和生化抗性机制。西花蓟马甲维盐抗性种群对阿维菌素有高水平交互抗性,抗性倍数为31.656,对啶虫脒有中等水平交互抗性,为12.182,对吡虫啉、溴虫腈、氯氟氰菊酯、毒死蜱和灭多威有低水平交互抗性,为5.517~8.568,而对多杀菌素无明显交互抗性。增效剂胡椒基丁醚(PBO)、马来酸二乙酯(DEM)、三丁基三硫磷酸酯(DEF)和磷酸三苯酯(TPP)对甲维盐抗性种群和田间种群均有显著增效作用。甲维盐抗性种群多功能氧化酶细胞色素P₄₅₀和b₅含量、O-脱甲基酶、谷胱甘肽S-转移酶和羧酸酯酶活性均显著提高,分别为敏感种群的3.89、3.61、5.32、4.42和1.30倍,表明多功能氧化酶、谷胱甘肽S-转移酶和羧酸酯酶等解毒代谢酶活性的提高是西花蓟马对甲维盐产生抗性的重要机制。     

小菜蛾对阿维菌素的抗性机制及交互抗性研究

用叶片药膜法研究了阿维菌素抗性小菜蛾 Plutella xylostella （L.）品系 对常用药剂的交互抗性谱以及增效醚（PB）和磷酸三苯酯（TPP）的增效作用。小菜蛾对阿 维菌素与高效氯氰菊酯、溴氰菊酯、氰戊菊酯和联苯菊酯等菊酯类药剂间具有比较低的交互 抗性，对后者抗性为3～20倍，对阿维菌素的抗性为575.6倍；对氟虫脲和氟啶脲没有交互抗 性。PB和TPP对阿维菌素分别增效8.2和5.5倍，说明小菜蛾对阿维菌素的抗性可能与多功能 氧化酶（MFO）和羧酸酯酶有关。通过差光谱技术测定了阿维菌素抗性和敏感小菜蛾细胞色 素P450的含量，抗性品系是敏感品系的1.38倍。     

褐飞虱对氟啶虫胺腈的抗性监测与生化抗性机制

为明确我国褐飞虱田间种群对氟啶虫胺腈的抗性现状及生化抗性机制,2017年-2019年采用稻茎浸渍法测定了采集自7省共13个褐飞虱田间种群对氟啶虫胺腈的抗性,并研究了氟啶虫胺腈抗性种群与其他杀虫剂的交互抗性以及增效剂对氟啶虫胺腈的增效效果。结果表明：近3年来褐飞虱对氟啶虫胺腈产生了中等水平抗性(RR=10.3～30.9)。氟啶虫胺腈抗性品系对呋虫胺、噻虫嗪和烯啶虫胺分别产生了9.1倍、7.9倍和4.1倍的低水平交互抗性,与噻嗪酮、毒死蜱、吡蚜酮、三氟苯嘧啶和吡虫啉不存在交互抗性。增效剂PBO对氟啶虫胺腈抗性品系和浙江龙游19(Longyou-19)田间种群分别具有4.2倍和3.8倍的明显增效作用。综上,褐飞虱田间种群已对氟啶虫胺腈产生中等水平抗性。多功能氧化酶参与了褐飞虱对氟啶虫胺腈的代谢抗性。     

小菜蛾对阿维菌素B_1抗药性选育及交互抗性

用阿维菌素B_1(abamectin)对小菜蛾敏感种群在室内进行抗性品系选育。经过25代连续汰选,获得抗性种群Laba-R,与选育前比较,抗性提高100倍。Laba-R种群在不接触任何药剂条件下饲养20代,抗性逐渐下降,很难恢复到选育前的敏感状态。抗性汰选前后分别测定10种药剂的剂量-死亡率毒力回归线,发现Laba-R抗性种群对乙酰甲胺磷、锐劲特、灭多威、敌敌畏不存在交互抗性;对溴氰菊酯、氯氰菊酯、杀虫双、巴丹和Bt的敏感性略有下降,但无明显交互抗性。活体增效剂试验表明,增效醚(PBO)和磷酸三苯酯(TPP)对阿维菌素B_1均有明显的增效作用,其中PBO的增效活性尤为显著,它能使对阿维菌素B_1产生100多倍抗药性的小菜蛾完全恢复其敏感性。说明多功能氧化酶解毒代谢增强可能是小菜蛾对阿维菌素B_1产生抗性的主导因素之一。     

小菜蛾对阿维菌素B1抗药性选育及交互抗性

用阿维菌素B1（abamectin)对小菜蛾敏感种群在室内进行抗性品系选育。经过25代连续汰选，获得抗性种群Laba-R，与选育前比较，抗性提高100倍，La-ba-R种群在不接触任何药剂条件下饲养20代，抗性逐渐下降，很难恢复到选育前的敏感状态。抗性汰选前后分别测定10种药剂的剂量-死亡率毒力回归线，发现Laba-R抗性种群对乙酰甲胺磷、锐劲特、灭多威、敌敌畏不存在交互抗性；对溴氰菊酯、氯氰菊酯、杀虫双、巴丹和Bt的第三性略有下降，但无明显交互抗性。活性增效剂试验表明，增效醚（PBO）和磷酸三苯酯（TPP）对阿维菌素B1均有明显的增效作用，其中PBO的增效活性尤为显著，它能使对阿维菌素B1产生100多倍抗药性的小菜蛾完全恢复其敏感性，说明多功能氧化酶解毒代谢增强可能是小菜蛾对阿维菌素B1产生抗性的主导因素之一。     

棉蚜对吡虫啉抗性的初步研究

用吡虫啉对棉蚜进行室内抗性筛选,用药处理25次后抗性是筛选前的20.03倍;2007年对田间棉蚜进行抗性调查,发现不同地区种群对吡虫啉的抗性差异显著,江苏南京种群最为敏感,河南安阳、山东泰安和北京地区棉蚜与之相比,抗性分别为2.21、7.63和9.53倍;抗、感品系解毒酶活力分析发现,抗性品系的谷胱甘肽S-转移酶活性增加很少(比活力1.12倍),但酯酶活力显著高于敏感品系(比活力1.71倍);增效试验结果表明,顺丁烯二酸二乙酯(DEM)在抗、感品系中对吡虫啉均没有明显的增效作用,而磷酸三苯酯(TPP)和增效醚(PBO)虽然在敏感品系中对吡虫啉的增效作用较小(SR 1.24和1.29),但在抗性品系中的增效作用显著增高(SR 2.13和1.74);此外还发现,吡虫啉处理可提高棉蚜群体的酯酶活力。由此认为,棉蚜至少具有对吡虫啉产生中等水平抗性的风险,其抗性可能是由于棉蚜的酯酶和P450单加氧酶的解毒能力提高所致。     

小菜蛾对阿维菌素的抗性选育及交互抗性研究

用阿维菌素对小菜蛾进行室内抗性汰选,选择压为约杀死种群70％的剂量。连续施药7～8次即表现抗性趋势,汰选至11代,获得抗性指数为80.71的抗性种群(ABM-R)。ABM-R种群对所测试的有机磷杀虫剂乙酰甲胺磷、有机氯类的硫丹、沙蚕毒素类的杀螟丹、氨基甲酸酯类的灭多威、微生物杀虫剂Bt、磺胺脲类衍生物丁醚脲、芳基取代的吡咯杂环化合物虫螨腈和昆虫生长调节剂定虫隆、虫酰肼无交互抗性(R/S为0.73～1.19),而对苯基吡唑类氟虫腈的敏感性却有所上升(R/S为0.22)。增效剂试验显示,PB和唧对ABM-S种群无增效作用,而对ABM-R种群增效作用显著,增效比分别为24.57和13.61,表明多功能氧化酶和羧酸酯酶在抗性机制中可能起重要作用。     

云南省小菜蛾田间种群对氯虫苯甲酰胺的抗药性变化趋势

为掌握云南省不同菜区小菜蛾种群对氯虫苯甲酰胺的抗药性水平及年度变化规律,于2009—2013年间连续5年,采用浸叶法监测了滇中通海县、滇西弥渡县和滇西南临沧市小菜蛾田间种群对氯虫苯甲酰胺的敏感性;同时,对通海小菜蛾田间种群的F1代,测定了磷酸三苯酯(TPP)、增效醚(PBO)和顺丁烯二酸二乙酯(DEM)3种增效剂对氯虫苯甲酰胺的增效作用。结果表明:自2009年以来,通海小菜蛾种群的抗药性水平持续上升,抗性倍数从2009年的2.30倍,逐渐发展到2012年的47.79倍和2013年的43.05倍,达到中等抗性水平;弥渡小菜蛾种群的抗药性水平除了2010年达12.35倍外,其余均处于低水平抗性;临沧市小菜蛾种群的抗药性水平在1.01~4.60倍之间,处于低水平抗性。3种增效剂均对氯虫苯甲酰胺无显著增效作用。     

武汉地区小菜蛾对溴氰菊酯的抗性回复及交互抗性

与小菜蛾敏感品系相比,武汉市张家湾小菜蛾田间种群的F₁、F₄和F₇,代对溴氰菊酯的抗性指数分别为362.79、30.78和10.91倍,抗性回复显著。用浸叶法测定Bt、抑太保和杀虫双制剂对两个小菜蛾种群的毒力,证明小菜蛾对这3类药剂与溴氰菊酯间无交互抗性。研究结果可为小菜蛾抗药性治理方案的制订提供依据。     

低剂量乙基多杀菌素对小菜蛾解毒酶的影响

为探讨低剂量乙基多杀菌素对小菜蛾Plutella xylostella（L.）解毒酶的影响,采用叶片浸渍法,测定了乙基多杀菌素和多杀菌素对小菜蛾敏感种群的毒力,并比较了低剂量（LC25和LC50）处理6、12、24、48和72 h时小菜蛾体内羧酸酯酶（CarE）、谷胱甘肽S-转移酶（GST）和多功能氧化酶系（MFOs）活性的变化动态。结果表明:乙基多杀菌素对小菜蛾的杀虫活性优于多杀菌素,处理48 h后其LC25和LC50浓度分别为0.018和0.048 mg/L,经此低剂量浓度处理后,小菜蛾CarE活性波动较大,6~24 h,处理组CarE活性高于对照组,且均呈先升后降趋势,24~72 h,处理组CarE活性均低于对照组,并且具有一定的时间效应;对GST具有明显的诱导作用,GST活性均高于对照组;对MFOs具有明显的抑制作用,除在48 h时相差不大外,其他时间MFOs活性均显著低于对照组。结果表明,GST可能参与了乙基多杀菌素在小菜蛾体内的代谢。     

Examination of stability of resistance and cross-resistance patterns to acylurea insect growth regulators in field populations of the diamondback moth,Plutella xylostella,from Malaysia

In teflubenzuron and teflubenzuron/chlorfluazuron laboratory-selected sub-populations of a Cameron Highlands (1988) strain of Plutella xylostella L., resistance to teflubenzuron reached much greater levels than chlorfluazuron but declined at a correspondingly greater rate to give similar resistance levels after 10–13 generations. There was little evidence of cross-resistance to flufenoxuron and none to hexaflumuron in these populations. Reselection of a Cameron Highlands population with teflubenzuron after 22 generations resulted in a significant (P < 0.05) increase in resistance after three generations. In two more recently collected strains from lowland Malaysia, Sawi (1991) and Serdang (1992) high levels of resistance to teflubenzuron were found, but only the former showed any apparent resistance to chlorfluazuron. There was also evidence of resistance to diflubenzuron in the Sawi strain (Serdang not examined). Resistance to teflubenzuron appeared to be unstable in unselected sub-populations of both lowland strains. Selection of the Sawi strain with teflubenzuron did not effect cross-resistance to chlorfluazuron, while attempted selection with chlorfluazuron had no effect on resistance to either compound. Selection of the Sawi strain with diflubenzuron increased resistance to teflubenzuron but had no effect on resistance to chlorfluazuron. In contrast to the Sawi strain, selection of the Serdang strain with teflubenzuron resulted in cross-resistance to chlorfluazuron. Synergist experiments with piperonyl butoxide (PB) and S,S,S-tributyl phosphorotrithioate (DEF) suggested that resistance to teflubenzuron in the lowland strains may involve both microsomal monooxygenases and esterases. The ovicidal activity of teflubenzuron, chlorfluazuron, hexaflumuron and flufenoxuron was consistently less than their activity against second-instar larvae, hexaflumuron, followed by teflubenzuron, showing the greatest relative ovicidal activity. Hexaflumuron was markedly more active than teflubenzuron against eggs from acylurea-selected Cameron Highlands populations and there was some evidence that resistance to teflubenzuron may be expressed at a lower level in eggs than in larvae in one such strain.     

烟粉虱对螺虫乙酯的抗性监测及交互抗性测定

为明确螺虫乙酯在湖北地区的抗性水平以及与其存在潜在交互抗性风险的常用药剂类型,采用着卵叶片浸渍法,于2013年对湖北省13个烟粉虱MEAM1和MED隐种地理种群对螺虫乙酯的抗药性进行了监测,并以室内敏感品系SUD-S为参照,对经螺虫乙酯连续17代抗性筛选的WH-2种群进行了交互抗性分析。结果表明:湖北省13个烟粉虱地理种群对螺虫乙酯均表现出极低的抗性,LC50介于0.015~0.081 mg/L之间;其中MED隐种WH-2种群对螺虫乙酯的抗性最高,LC50为0.081 mg/L。在室内用螺虫乙酯对MED隐种WH-2种群连续17代抗性筛选后,其对螺虫乙酯的抗性倍数上升约7倍。经螺虫乙酯筛选的WH-2种群对吡虫啉、乙酰甲胺磷、灭多威、氟氯氰菊酯、氰戊菊酯和氟啶虫胺腈均产生了低水平的交互抗性,抗性倍数分别为8.699、7.165、5.317、6.681、2.958、5.662倍,而对毒死蜱和阿维菌素无交互抗性。表明烟粉虱在湖北省部分地区对螺虫乙酯存在低水平抗性,且螺虫乙酯与部分常用杀虫剂存在交互抗性风险。     

Metabolic resistance mechanisms to phosalone in the common pistachio psyllid, Agonoscena pistaciae (Hem.: Psyllidae)

The common pistachio psyllid, Agonoscena pistaciae, is the most damaging pest of pistachio in Iran, and is generally controlled by insecticides belonging to various classes especially, phosalone. The toxicity of phosalone in nine populations of the pest was assayed using the residual contact vial and insect-dip methods. The bioassay results showed significant discrepancy in susceptibility to phosalone among the populations. Resistance ratio of the populations to the susceptible population ranged from 3.3 to 11.3. The synergistic effects of TPP, PBO and DEM were evaluated on the susceptible and the most resistant population to determine the involvement of esterases, mixed function oxidases and glutathione S-transferases in resistance mechanisms, respectively. The level of resistance to phosalone in the resistant population was suppressed by TPP, PBO and DEM, suggesting that the resistance to phosalone is mainly caused by esterase detoxification. Biochemical enzyme assays revealed that esterase, glutathione S-transferase and cytochrome P450 monooxygenase activities in the resistant population was higher than that in the susceptible. Glutathione-S-transferases play a minor role in the resistance of the pest to phosalone.     

Synergism of teflubenzuron and chlorfluazuron in an acylurea-resistant field strain of Plutella xylostella L. (lepidoptera: Yponomeutidae)

Topical application of the synergists piperonyl butoxide (PB) and S,S,S-tributyl phosphorotrithioate (DEF) to second-instar larvae of a standard laboratory strain (FS) and an unselected Malaysian field strain (CH) of the diamondback moth Plutella xylostella had no significant effect on the toxicity of the acylurea insecticides, chlorfluazuron and teflubenzuron, in a subsequent leafdip bioassay. In contrast, pre-treatment with PB or DEF in acylurea-selected subpopulations of the CH strain with varying levels of cross-resistance to chlorfluazuron and teflubenzuron significantly increased (up to 34-fold and 28-fold, respectively) the toxicity of both compounds, suggesting that microsomal monooxygenases and esterases may be involved in resistance. The addition of a mineral oil, ‘Sunspray 6E’, to topically-applied chlorfluazuron consistently reduced its LD₅₀ value, and the effect of the oil appeared to be greatest on the most resistant population of P. xylostella. However, the effects of the oil were not significant (P > 0·05) and further studies are necessary to determine whether a penetration factor is present in the CH strain.     

Biochemical mechanisms conferring cross-resistance between tebufenozide and abamectin in Plutella xylostella

Experiments have been carried out to confirm the cross-resistance between abamectin and tebufenozide in Plutella xylostella and demonstrate its mechanism. The results showed that the resistant strain of P. xylostella selected by tebufenozide (RF 99.38) really showed high cross-resistance to abamectin (RF 29.25). When this strain was subjected to resistance decaying treatment, breeding without contacting any insecticides, and abamectin resistance selection for 20 generations, the former resulted in decrease of its resistance to both tebufenozide and abamectin to about one third of the original (RF 35.03 and 11.67, respectively), and the later enhanced its resistance to abamectin dramatically (RF 303.77), but not to tebufenozide(RF 50.04). PBO showed high synergism to abamectin (SR 2.11-12.23), and the synergism ratio positively related to the resistance level among different strains. Enzyme analysis also proved that the activity of cytochrome P450 monooxygenase (MFO) was notable enhanced in the strains resistant to both tebufenozide and abamectin (1.71- to 3.01-fold). Based on discussion, it was concluded that tebufenozide selection could resulted in significant cross-resistance of P. xylostella to abamectin. The major mechanism for the cross-resistance should be the enhancement of MFO activity. For resistance management, tebufenozide and abamectin would not recommend for rotational use.     

河南省麦田荠菜对苯磺隆的抗性及其交互抗性

为明确河南省荠菜Capsella bursa-pastoris种群对苯磺隆的抗性水平及其可能存在的抗性机理,应用整株法测定了采自驻马店及南阳等6个荠菜发生严重市的10个荠菜种群对苯磺隆的抗性,扩增和比对了荠菜苯磺隆抗性种群及敏感种群之间靶标酶乙酰乳酸合成酶基因ALS的差异,并使用单剂量法测定了以上种群对双氟磺草胺、啶磺草胺及氟唑磺隆等ALS抑制剂类除草剂的交互抗性。结果表明,驻马店市的汝南县冯湾村(ZMD-1)及平舆县五里路村(ZMD-3)荠菜种群对苯磺隆的抗性倍数分别为3.1和2.5,表现出低水平抗性;驻马店市汝南县赖楼村(ZMD-2)和周口市川汇区文庄村(ZK-1)荠菜种群对苯磺隆的抗性倍数分别为21.7和57.8,表现出高水平抗性;南阳市唐河县上屯村(NY-2)荠菜种群对苯磺隆的抗性倍数为116.5,表现出极高水平抗性,其它种群对苯磺隆仍然较敏感。NY-2、ZMD-2和ZK-1种群的ALS基因第197位氨基酸由脯氨酸(CCT)分别突变为丝氨酸(TCT)、丙氨酸(GCT)和亮氨酸(CTT),其它种群中均未发现有突变产生;这3个种群在氟唑磺隆推荐剂量处理下,死亡率仅为18.9%、23.3%和11.1%,说明已对氟唑磺隆产生了较高水平的交互抗性,其中NY-2种群对双氟磺草胺和啶磺草胺产生了低水平交互抗性,推荐剂量下死亡率分别为82.2%和83.1%。表明ALS基因突变很可能是导致荠菜种群对苯磺隆等ALS抑制剂类除草剂产生抗性的重要原因。     

Cross-resistance and biochemical mechanisms of abamectin resistance in the western flower thrips, Frankliniella occidentalis

Abamectin resistance was selected in the western flower thrips [Frankliniella occidentalis (Pergande)] under the laboratory conditions, and cross-resistance patterns and possible resistance mechanisms in the abamectin-resistant strain (ABA-R) were investigated. Compared with the susceptible strain (ABA-S), the ABA-R strain displayed 45.5-fold resistance to abamectin after 15 selection cycles during 18 generations. Rapid reversion of abamectin resistance was observed in the ABA-R strain in the absence of the insecticide selection pressure. Moderate and low levels of cross-resistance to chlorpyrifos (RR 11.4) and lambda-cyhalothrin (3.98) were observed in the ABA-R strain, but no significant cross-resistance was found to spinosad (2.00), acetamiprid (1.47) and chlorfenapyr (0.26). Our studies also showed that the esterase inhibitor S,S,S-tributyl phosphorotrithioate (DEF) and glutathione S-transferase inhibitor diethyl maleate (DEM) were not able to synergize the toxicity of abamectin, whereas the oxidase inhibitor piperonyl butoxide (PBO) conferred a significant synergism on abamectin in the ABA-R strain (SR 3.00). Biochemical analysis showed that cytochrome P450 monooxygenase activity of the ABA-R strain was 6.66-fold higher than that of the ABA-S strain. It appears that enhanced oxidative metabolism mediated by cytochrome P450 monooxygenases was a major mechanism for abamectin resistance in the western flower thrips.     

四种P450基因调控小菜蛾对氯虫苯甲酰胺的抗性

为研究CYP6家族P450基因在小菜蛾Plutella xylostella代谢氯虫苯甲酰胺中的作用,利用浸叶法测定不同小菜蛾种群3龄幼虫对氯虫苯甲酰的抗性水平,采用实时荧光定量PCR(quantitative realtime PCR,q RT-PCR)方法分析CYP1v3、CYP1v4、CYP6B6和CYP6f这4种P450基因在小菜蛾不同抗性种群体内的表达差异及杀虫剂的短期诱导效应,并通过RNA干扰技术沉默4种P450基因后分析小菜蛾对氯虫苯甲酰胺的敏感性。结果显示,4种P450基因在中等抗性水平小菜蛾种群体内高表达;氯虫苯甲酰胺处理可诱导4种基因显著上调表达。分别沉默4种P450基因后,处理组小菜蛾的P450酶活力显著下降37.60%～54.82%,且处理组小菜蛾的死亡率显著高于对照组;同时沉默4种基因处理组的小菜蛾P450酶活力显著低于其他各处理组。表明4种P450基因可能同时参于小菜蛾对氯虫苯甲酰胺的代谢。