抗吡虫啉棉蚜对其他新烟碱类药剂的交互抗性及相关酶的活性变化

为明确抗吡虫啉棉蚜Aphis gossypii对其他新烟碱药剂交互抗性及相关酶活性的变化, 本研究以室内筛选的棉蚜吡虫啉抗性种群、山东夏津县田间多抗种群和敏感种群为材料,采用生测法测定了这3种不同抗性种群棉蚜对呋虫胺和烯啶虫胺的交互抗性,以及3种解毒酶抑制剂对吡虫啉等药剂的增效作用,采用生化分析法测定了这3个种群棉蚜解毒酶及乙酰胆碱酯酶的活性和药剂的抑制作用。结果表明：吡虫啉抗性种群、夏津田间多抗种群的棉蚜对呋虫胺均无交互抗性,但对烯啶虫胺的交互抗性分别达5.28和4.89倍。呋虫胺对抗吡虫啉棉蚜的羧酸酯酶、乙酰胆碱酯酶及谷胱甘肽-S-转移酶都有显著的抑制作用;烯啶虫胺对抗吡虫啉棉蚜的羧酸酯酶、乙酰胆碱酯酶及谷胱甘肽-S-转移酶的抑制作用较小。羧酸酯酶抑制剂TPP和多功能氧化酶抑制剂PBO对吡虫啉和烯啶虫胺都有明显的增效作用,但对呋虫胺的增效作用不明显; 谷胱甘肽-S-转移酶抑制剂DEM对3种药剂均没有明显的增效作用。呋虫胺和烯啶虫胺都可以抑制抗吡虫啉棉蚜的解毒酶及乙酰胆碱酯酶,其中以呋虫胺的抑制效果较为显著。结果显示呋虫胺在抗吡虫啉棉蚜治理中的应用价值较大,其结构可为今后新烟碱类药剂的开发提供参考。     

棉蚜啶虫脒抗性种群交互抗性和增效剂增效作用的研究

【目的】明确棉蚜Aphis gossypii Glover啶虫脒抗性品系与其它杀虫剂的交互抗性现状以及增效剂的增效作用,为延缓和治理棉蚜对啶虫脒的抗性提供依据。【方法】采用单头反选育和群体汰选的方式,获得了棉蚜啶虫脒敏感和抗性品系;采用叶片药膜法测定了13种杀虫剂对啶虫脒的交互抗性以及增效剂对啶虫脒的增效作用。【结果】经过室内棉蚜敏感和抗性品系的筛选,获得了相对抗性倍数为82.33倍的棉蚜啶虫脒抗性品系。棉蚜啶虫脒抗性品系的交互抗性谱的研究表明,交互抗性倍数小于5的药剂为:吡蚜酮,甲基阿维菌素;交互抗性倍数在5~10倍的药剂为:噻虫嗪,联苯菊酯,毒死蜱,马拉硫磷,丙溴磷,辛硫磷;交互抗性倍数在10~15倍的药剂为:硫丹,阿维菌素,高效氯氰菊酯,三唑磷,氧化乐果;交互抗性倍数大于1 5倍的药剂为:吡虫啉。增效剂实验表明,TPP和PBO在啶虫脒敏感品系中增效作用不明显,但在抗性品系中增效作用显著。在啶虫脒抗性品系中的增效比为1.77、1.61,在啶虫脒敏感品系中的增效比为1.02、1.03。DEM在啶虫脒抗性、敏感品系中的增效作用均不明显,增效比为1.04、1.02。TPP和PBO对啶虫脒有很好的增效作用。以室内棉蚜敏感品系(LC_(50)为0.180 mg/L)为基础,对新疆各主要棉区的棉蚜种群进行了啶虫脒药剂的抗性调查,结果表明新疆各主要棉区棉蚜对啶虫脒的相对抗性倍数为6.1~22.0倍。【结论】由此说明新疆主要棉区棉蚜对啶虫脒具有一定的抗性风险,生产中可以利用无交互抗性的吡蚜酮和甲基阿维菌素来治理抗性棉蚜种群。     

我国棉花主产区棉蚜对吡虫啉的抗性监测及抗性机理

【目的】由于生长周期短、繁殖率高,棉蚜Aphis gossypii容易对杀虫剂产生抗药性。本研究旨在明确我国棉花主产区棉蚜对吡虫啉的抗性水平及抗性机理。【方法】采用浸叶法测定了北京海淀,河北廊坊和邯郸,山东德州,河南许昌,以及新疆奎屯和阿克苏地区棉蚜对吡虫啉的抗性水平;测定了不同种群棉蚜3种解毒酶（多功能氧化酶、羧酸酯酶、谷胱甘肽S-转移酶）及乙酰胆碱酯酶的活性;并对靶标基因烟碱型乙酰胆碱受体（nAChR）β1亚基基因进行了突变检测。【结果】北京海淀、河南许昌和河北邯郸的棉蚜对吡虫啉敏感;河北廊坊、新疆阿克苏、山东德州及新疆奎屯地区的棉蚜对吡虫啉的抗性倍数(resistance ratio, RR)分别为22.6, 26.3,53.5和61.1倍,为中等水平抗性。酶活力对比研究发现,阿克苏和奎屯地区的棉蚜多功能氧化酶的比活力分别是敏感种群(北京种群)的1.7和1.8倍,羧酸酯酶的比活力分别是敏感种群的1.6和1.7倍,谷胱甘肽S-转移酶的比活力均是敏感种群的1.5倍,但是乙酰胆碱酯酶比活力在棉蚜种群间差异不显著。靶标基因突变检测表明,河北廊坊、新疆阿克苏、山东德州及新疆奎屯棉蚜种群nAChR β1亚基均存在与吡虫啉抗性相关的精氨酸到苏氨酸（R81T）突变。【结论】结果提示,多功能氧化酶、羧酸酯酶和谷胱甘肽S-转移酶活力升高以及nAChR β1亚基R81T突变与棉蚜对吡虫啉的抗性形成相关。     

西花蓟马抗噻虫胺种群对杀虫剂的交互抗性及机制

噻虫胺是具有内吸和触杀等多种作用方式的新烟碱类杀虫剂,常用于防治入侵害虫西花蓟马。为明确抗性风险,本文研究了西花蓟马抗噻虫胺种群对多种杀虫剂的交互抗性及其机制。经过45代筛选,西花蓟马对噻虫胺产生了高水平抗性(56.8倍)。生物测定结果表明: 西花蓟马高抗噻虫胺种群与噻虫嗪、吡虫啉、毒死蜱、三氟氯氰菊酯、甲维盐存在中等水平交互抗性(18.6>RR₅₀>11.3),对辛硫磷及灭多威具有低水平交互抗性,与溴虫腈和多杀菌素不存在交互抗性。胡椒基丁醚(PBO)与磷酸三苯酯(TPP)对杀灭西花蓟马抗噻虫胺种群(CL)、云南田间种群(YN)和敏感种群(S)均有显著增效作用。西花蓟马抗噻虫胺种群细胞色素P₄₅₀含量(3.6倍)、细胞色素b₅含量(2.9倍)及O-脱甲基酶活性(4.9倍)和羧酸酯酶活性(2.5倍)均显著高于敏感种群,表明多功能氧化酶及羧酸酯酶活性增强是西花蓟马对噻虫胺产生抗性的重要机制。     

西花蓟马对烯啶虫胺、噻虫胺和噻虫嗪的抗性风险和抗性稳定性

为明确使用新烟碱类杀虫剂烯啶虫胺、噻虫胺和噻虫嗪防治入侵害虫西花蓟马的抗性风险及抗性稳定性,本研究采用芸豆浸药法对西花蓟马敏感种群初羽化雌成虫进行连续筛选获得抗性种群,根据抗性现实遗传力计算公式分析西花蓟马对上述3种杀虫剂的抗性风险,预测其抗性发展速度,并测定抗性稳定性。结果表明: 经过30代抗性筛选,西花蓟马对烯啶虫胺、噻虫胺和噻虫嗪均达到高水平抗性(44.7、45.5和32.7倍)。西花蓟马对噻虫胺、烯啶虫胺和噻虫嗪的抗性发展速度依次降低,抗性现实遗传力分别为0.1503、0.1336和0.1258。对抗性种群在无选择压力下继续饲养10代,西花蓟马对烯啶虫胺、噻虫胺和噻虫嗪的抗性水平均出现一定程度的下降,但均未能恢复到敏感性水平。抗性选育后,西花蓟马若虫与成虫对杀虫剂的敏感性差异显著缩小,西花蓟马敏感种群及抗性种群若虫对上述3种杀虫剂的敏感性显著高于成虫。西花蓟马对烯啶虫胺、噻虫胺和噻虫嗪均存在高抗风险,噻虫嗪的抗性上升速度较慢且抗性稳定性最低。因此,在西花蓟马若虫期使用噻虫嗪有利于西花蓟马防治。     

贵州稻区褐飞虱种群对六种杀虫剂的抗性动态

【目的】为明确贵州褐飞虱Nilaparvata lugens种群对常用杀虫剂的抗性水平。【方法】在室内采用浸渍法测定了2013-2015年采自贵州黄平、桐梓和开阳3地的褐飞虱种群对6种杀虫剂(吡虫啉、噻虫嗪、异丙威、噻虫胺、啶虫脒和醚菊酯)的抗性。【结果】贵州褐飞虱种群对不同的杀虫剂存在着抗性差异。与敏感品系相比,2013-2015年间3地田间褐飞虱种群对吡虫啉、噻虫嗪、异丙威、噻虫胺的抗性均达到中等水平抗性,抗性倍数(resistance ratio,RR)分别为21.88~95.38,10.91~69.36,13.00~57.23和23.11~39.54倍;而对啶虫脒和醚菊酯仍处于敏感阶段,RR分别为0.47~0.75和0.41~0.85倍。【结论】褐飞虱对吡虫啉和噻虫嗪的抗性较高,可能与近年来广泛地大量使用有关。本研究的抗性动态监测结果对贵州稻区褐飞虱的杀虫剂种类调整及施药策略等具有重要指导作用。     

抗吡虫啉棉蚜种群对吡蚜酮等药剂的交互抗性及施药对其生物学特性的影响

为了明确吡蚜酮对抗吡虫啉棉蚜Aphis gossypii种群的防治效果,提出抗吡虫啉棉蚜的治理策略,利用抗吡虫啉棉蚜种群(RF₂₇)、敏感种群(SS)和夏津田间种群(XJ),分别采用浸渍法、微量点滴法、生化测定法和系统观察法研究了棉蚜无翅成蚜对吡蚜酮等药剂的交互抗性及施药对其生物学特性的影响。结果表明: 吡蚜酮对RF₂₇,XJ和SS的LD₅₀分别为1.213×10^-5,8.506×10^-5和5.140×10^-5 μg/头,RF₂₇对吡蚜酮表现出明显的负交互抗性现象。RF₂₇对啶虫脒、烯啶虫胺和噻嗪酮分别产生2.35,2.98和1.71倍的抗性。RF₂₇的羧酸酯酶和谷胱甘肽-S-转移酶的比活力较SS分别高2.73和1.57倍,说明羧酸酯酶和谷胱甘肽-S-转移酶比活力的提高是引起棉蚜对吡虫啉产生抗性的重要原因之一。吡蚜酮分别处理RF₂₇,XJ和SS,其羧酸酯酶和谷胱甘肽-S-转移酶均受到显著抑制。吡蚜酮以低剂量处理RF₂₇成蚜,对其生长发育有显著的不利影响,表现为若蚜存活率降低(64.60%),净生殖率降低（10.39）,内禀增长率和周限增长率显著降低(分别为0.21和1.23),世代历期延长（10.87 d）,相对适合度较小（仅为0.70）。这些结果表明吡蚜酮在棉蚜防治中具有很高的应用价值。     

棉蚜对吡虫啉的抗性选育和现实遗传力分析

【目的】为了评估棉蚜Aphis gossypii Glover对吡虫啉的抗性风险,在室内进行了棉蚜对吡虫啉(imidacloprid)的抗性选育和抗性现实遗传力分析。【方法】采用单头反选育法和群体汰选法,分别得到了棉蚜对吡虫啉敏感品系(LC₅₀为0.176 mg/L)和抗性品系(LC₅₀为14.657 mg/L)。采用阈性状分析方法,获得棉蚜对吡虫啉的抗性现实遗传力（h²）。【结果】相对于田间原始种群(LC₅₀为0.346 mg/L),吡虫啉敏感棉蚜品系对吡虫啉的LC50减少了2倍;获得的吡虫啉抗性棉蚜品系,经过40代的选育,得到抗性倍数为室内敏感品系的83.27倍的抗性品系。棉蚜对吡虫啉的抗性现实遗传力(h2)为0.1478。进一步预测其抗性发展速度,基于80%~90%的选择压力,预计抗性增长100倍时,吡虫啉可使用30.2~38.1代。【结论】这些研究说明棉蚜对吡虫啉存在抗性风险。     

六种新烟碱类杀虫剂对凹唇壁蜂的毒性及风险评估

【目的】为评价新烟碱类杀虫剂对凹唇壁蜂Osmia excavata的毒性和风险性。【方法】分别采用饲喂法和接触法测定了啶虫脒、噻虫嗪、氟啶虫胺腈、吡虫啉、噻虫胺、呋虫胺6种新烟碱类杀虫剂对凹唇壁蜂雌成蜂的24 h和48 h的急性经口和接触毒性。并依据农药对蜜蜂生态风险的危害熵值(hazard quotient,HQ)评估其对凹唇壁蜂的风险性。【结果】经口毒性测定中,氟啶虫胺腈对凹唇壁蜂雌蜂毒性最高,噻虫嗪毒性最低;接触毒性测定中,噻虫胺对凹唇壁蜂雌蜂毒性最高,氟啶虫胺腈毒性最低。啶虫脒、噻虫嗪、吡虫啉对凹唇壁蜂表现为中等风险;噻虫胺经对凹唇壁蜂表现为高风险。氟啶虫胺腈和呋虫胺经接触途径表现为中等风险,而经口途径表现为高风险。【结论】本研究所选的6种新烟碱类杀虫剂对凹唇壁蜂雌成蜂的毒性均为高毒。在果园中应尽量避免使用噻虫胺、氟啶虫胺腈及呋虫胺,在植物花期谨慎使用啶虫脒、噻虫嗪、吡虫啉等新烟碱类杀虫剂。     

褐飞虱对噻虫嗪和烯啶虫胺的抗性风险评估

采用稻茎浸渍法,测定了广州市本地褐飞虱种群对噻虫嗪和烯啶虫胺的室内毒力,评估褐飞虱对其的抗性风险。结果表明：广州本地褐飞虱种群对噻虫嗪和烯啶虫胺的LC50分别为02857mg/L和05022 mg/L,分别是敏感品系LC50的267倍和106倍,仍属敏感水平。室内抗性筛选结果表明：经过30代的连续筛选后,噻虫嗪的抗性上升82980倍,达到极高抗性水平,烯啶虫胺的抗性上升3170倍,达到中等抗性水平,表明褐飞虱对噻虫嗪和烯啶虫胺存在抗性风险的可能。根据试验结果,对褐飞虱噻虫嗪和烯啶虫胺抗性的预防治理提出了应用策略。     

Resistance of aphis gossypii (Homoptera: Aphididae) to fenvalerate and imidacloprid and activities of detoxification enzymes on cotton and cucumber

Resistance of two strains of cotton aphid, Aphis gossypii Glover, to fenvalerate and imidacloprid were determined on cotton (Gossypium hirsutum L.) and cucumber (Cucumis sativa L.) after resistance selection of one strain to fenvalerate for 16 consecutive generations, and of a second strain to imidacloprid for 12 consecutive generations on cotton in greenhouses. Dose-response and activities of detoxication enzymes of the fenvalerate-resistant strain (R-fenvalerate), the imidacloprid-resistant strain (R-imidacloprid), and a susceptible strain (S) were determined. After 16 consecutive generations of selection, resistance of A. gossypii to fenvalerate increased >29,000-fold and to imidacloprid 8.1-fold. On cucumber. resistance of the R-fenvalerate strain to fenvalerate increased 700-fold and to imidacloprid 3.6-fold. However, the most significant finding in this study was that the R-imidacloprid strain exhibited cross-resistance to fenvalerate, with a resistance ratio of 108.9-fold on cotton and 3:3.5-fold on cucumber, whereas the R-fenvalerate strain did not show significant cross-resistance to imidacloprid on either plant species. Both resistant strains of A. gossypii were more resistant to fenvalerate on cotton than on cucumber, whereas their susceptibility to imidacloprid on otton and cucumber were not significantly different. The response of the S strain to fenvalerate and imidacloprid were similar on cotton and Cucumber. Activities of acetylcholinesterase (AChE) and alpha-naphthylacetate (alpha-NA) esterases of A. gossypii were significantly different among the three strains, with the R-fenvalerate strains having the highest, followed by the R-imidacloprid strain, and the S strain the lowest. The activities of the AChE and alpha-NA esterases for all three strains were also significantly higher on cotton than on cucumber. The resistance mechanism and resistance management strategies for the R-fenvalerate and R-imidacloprid strains of A. gossypii to fenvalerate and imidacloprid on cotton and cucumber are discussed.     

Assessment of cross-resistance potential to neonicotinoid insecticides in Bemisia tabaci (Hemiptera: Aleyrodidae)

Laboratory bioassays were carried out with four neonicotinoid insecticides on multiple strains of Bemisia tabaci (Gennadius) to evaluate resistance and cross-resistance patterns. Three imidacloprid-resistant strains and field populations from three different locations in the southwestern USA were compared in systemic uptake bioassays with acetamiprid, dinotefuran, imidacloprid and thiamethoxam. An imidacloprid-resistant strain (IM-R) with 120-fold resistance originally collected from Imperial Valley, California, did not show cross-resistance to acetamiprid, dinotefuran or thiamethoxam. The Guatemala-resistant strain (GU-R) that was also highly resistant to imidacloprid (RR=109-fold) showed low levels of cross-resistance when bioassayed with acetamiprid and thiamethoxam. However, dinotefuran was more toxic than either imidacloprid or thiamethoxam to both IM-R and GU-R strains as indicated by low LC50s. By contrast, a Q-biotype Spanish-resistant strain (SQ-R) of B. tabaci highly resistant to imidacloprid demonstrated high cross-resistance to the two related neonicotinoids. Field populations from Imperial Valley (California), Maricopa and Yuma (Arizona), showed variable susceptibility to imidacloprid (LC50s ranging from 3.39 to 115 microg ml(-1)) but did not exhibit cross-resistance to the three neonicotinoids suggesting that all three compounds would be effective in managing whiteflies. Yuma populations were the most susceptible to imidacloprid. Dinotefuran was the most toxic of the four neonicotinoids against field populations. Although differences in binding at the target site and metabolic pathways may influence the variability in cross-resistance patterns among whitefly populations, comparison of whitefly responses from various geographic regions to the four neonicotinoids indicates the importance of ecological and operational factors on development of cross-resistance to the neonicotinoids.     

华北棉区主要害虫抗药性监测与治理技术示范

【目的】华北地区转基因Bt棉大面积种植后,刺吸式口器害虫已成为棉花主要害虫。本研究旨在监测明确棉花主要害虫对田间常用杀虫剂抗性水平变化,以指导田间合理用药。【方法】2013-2015年分别采用叶片药膜法和点滴法系统监测了河北省邱县、山东省滨州市、河南省西华县棉铃虫Helicoverpa armigera、棉蚜Aphis gossypii、绿盲蝽Apolygus lucorum对常用杀虫药剂的抗性动态变化情况。【结果】棉铃虫对辛硫磷、高效氯氟氰菊酯抗性呈上升趋势,抗性倍数都在20倍以上,对甲氨基阿维菌素苯甲酸盐从敏感状态转为中等水平抗性,抗性倍数在10倍以上。棉蚜对杀虫剂抗性水平都比较高,特别是对氧化乐果、高效氯氰菊酯、吡虫啉都已产生了高水平抗性,抗性倍数都在100倍以上。绿盲蝽对吡虫啉从敏感状态转为中等水平抗性,抗性倍数在10倍以上,对马拉硫磷、灭多威等药剂抗性倍数还维持在10倍以下,对高效氯氟氰菊酯只监测到滨州种群产生了中等水平的抗性,抗性倍数达到了95倍。在明确棉花害虫抗药性水平的基础上,对山东滨州棉花害虫实施了以轮换用药为主的抗性治理示范,示范区比农户自防对照区减少3次用药,增加棉花产量7.53%,节本增收109.16元,取得了较好的示范效果和经济效益。【结论】当前华北棉区主要害虫抗性水平上升,急需开展以轮换用药为主的抗性治理措施。     

棉蚜抗吡虫啉品系和敏感品系主要解毒酶活性比较

通过生物测定和生物化学方法比较了棉蚜 Aphis gossypii 对吡虫啉抗性（约为7倍）和敏感品系几种主要解毒酶的活性。结果表明：氧化胡椒基丁醚对两个品系均无明显增效作用。抗性品系中羧酸酯酶和谷胱甘肽S-转移酶的比活力均明显高于敏感品系,抗性品系中羧酸酯酶的K_m值也显著高于敏感品系,说明抗性品系羧酸酯酶与底物的亲和力明显高于敏感品系。上述结果证明羧酸酯酶和谷胱甘肽S-转移酶活力增强在棉蚜对吡虫啉的抗性中起重要作用。     

八种新烟碱类杀虫剂对地熊蜂工蜂的毒性及风险评估

【目的】评价新烟碱类杀虫剂对地熊蜂Bombus terrestris工蜂的毒性和生态风险性,为温室施用新烟碱类杀虫剂提供科学依据。【方法】分别采用饲喂法和接触法测定了噻虫嗪、噻虫胺、啶虫脒、吡虫啉、烯啶虫胺、呋虫胺、噻虫啉和氟吡呋喃酮8种新烟碱类杀虫剂对地熊蜂成年工蜂的急性经口和急性接触毒性。同时评估了8种新烟碱类杀虫剂对地熊蜂工蜂的生态风险性。【结果】8种杀虫剂经饲喂法测定,噻虫胺对地熊蜂成年工蜂的毒性最高,24 h和48 h的LD 50值分别为0.0433和0.0330μg a.i./蜂;氟吡呋喃酮毒性最低,24 h和48 h的LD 50值分别为72.4119和67.9079μg a.i./蜂。接触法测定的毒性与饲喂法测得的结果一致,噻虫胺的毒性最高,24 h和48 h的LD 50值分别为0.0220和0.0192μg a.i./蜂;氟吡呋喃酮的毒性最低,24 h和48 h的LD 50值分别为141.7641和130.3062μg a.i./蜂。生态风险评估表明,啶虫脒、噻虫啉和氟吡呋喃酮对地熊蜂成年工蜂的经口毒性和接触毒性均表现为低风险,吡虫啉、烯啶虫胺和呋虫胺的毒性表现为中等风险。噻虫嗪和噻虫胺对地熊蜂成年工蜂的经口毒性表现为中等风险,而接触毒性则表现为高风险。【结论】检测的8种新烟碱类杀虫剂中啶虫脒、噻虫啉与氟吡呋喃酮对地熊蜂成年工蜂的毒性为低毒,而噻虫嗪、噻虫胺、吡虫啉、烯啶虫胺、呋虫胺这5种杀虫剂均为高毒。在设施蔬菜花期使用地熊蜂授粉时,建议禁用这5种中、高风险的新烟碱类杀虫剂,以避免对熊蜂授粉的危害,而另3种低风险药剂可根据田间试验情况合理施用。