斜纹夜蛾对茚虫威的抗性机制

以斜纹夜蛾对茚虫威相对敏感种群和抗性倍数为15.63倍的抗性选育种群为材料,通过解毒酶活性测定与钠离子通道基因片段的克隆、测序,研究了斜纹夜蛾对茚虫威的抗性机制.结果表明,与相对敏感种群相比,抗性种群酯酶活性提高了2.27倍,但抗性和敏感种群谷胱甘肽S-转移酶和多功能氧化酶O-脱甲基活性无显著差异;3龄幼虫酯酶活性随着抗性指数的上升而逐渐提高;从相对敏感和抗性斜纹夜蛾种群3龄幼虫基因组DNA中扩增出位于钠离子通道IIS5-IIS6的341 bp DNA片段;与相对敏感种群相比,茚虫威抗性种群钠离子通道基因没有发生突变.     

不同食料对甜菜夜蛾药剂敏感性及解毒酶活性的影响

甜菜夜蛾取食不同食料作物以后,对灭多威、辛硫磷和溴氰菊酯的敏感性均有不同程度的变化。解毒酶活性测定表明,不同食料对甜菜夜蛾3龄幼虫的多功能氧化酶(MFO)的脱甲基作用和羧酸酯酶(CarE)的活性有一定的诱导作用,和人工饲料相比,具有显著性差异。而羧酸酯酶动力学研究表明,几种食料作物对甜菜夜蛾羧酸酯酶底物亲和力(Km)的影响差异不显著。     

转Cry1Ac/1Ab基因棉花对棉铃虫的控制效果及对甜菜夜蛾和斜纹夜蛾生长发育的影响

为评估转Cry1Ac/1Ab基因棉花对棉铃虫Helicoverpa armigera的抗性及对非靶标害虫甜菜夜蛾Spodoptera exigua和斜纹夜蛾S.litura生长发育的影响,采用室内生测法测定其对棉铃虫的抗性及对甜菜夜蛾和斜纹夜蛾不同龄期幼虫存活率、营养代谢及中肠酶活性的影响。结果表明,转Cry1Ac/1Ab基因棉花对棉铃虫第2代幼虫的抗性程度最高,幼虫校正死亡率达91.33%,但对甜菜夜蛾和斜纹夜蛾的抗性程度较低,幼虫校正死亡率分别为15.33%和13.33%。甜菜夜蛾和斜纹夜蛾各龄期幼虫取食转Cry1Ac/1Ab基因棉花后,其存活率与取食常规棉花对照无显著差异;甜菜夜蛾对转Cry1Ac/1Ab基因棉花叶片的相对取食量、近似消化率分别为16.68和93.12%,均高于取食常规棉花对照的10.72和92.00%,但差异不显著,而斜纹夜蛾取食转Cry1Ac/1Ab基因棉花后的各项营养指标均低于取食常规棉花对照,差异也不显著。取食转Cry1Ac/1Ab基因棉花后,甜菜夜蛾的过氧化物酶活性为0.02 U/mg prot,显著低于取食常规棉花的0.05 U/mg prot;斜纹夜蛾的酸性磷酸酶活性为0.15 U/mg prot,高于取食常规棉花的0.10 U/mg prot,但差异不显著,其它中肠酶活性均低于对照,亦无显著差异。     

小菜蛾对杀虫双的抗性遗传研究

利用室内选育的敏感品系和抗杀虫双品系为亲本,采用剂量对数—死亡机率值回归线(LD-P线)分析法,研究了小菜蛾对杀虫双的抗性遗传方式。结果表明,小菜蛾对杀虫双的抗性为多基因、常染色体遗传,正、反交F_1的显性度(D)值分别为0.39、0.28,即其主效基因为不完全显性。小菜蛾对杀虫双的抗性现实遗传力较低,h~2=0.052,产生抗性的速率较慢,室内选育119代,抗性仅达122.8倍。抗杀虫双品系和遗传杂交后代(F_1、F_2、BC)对拟除虫菊酯类、氨基甲酸酯类、有机磷类的代表杀虫剂溴氰菊酯、灭多威、敌敌畏等的交互抗性测定结果表明,它们对3种杀虫剂无交互抗性;亲本和杂交后代的多功能氧化酶环氧化活性与杀虫双的抗性水平呈正相关性;乙酰胆碱酯酶活性要比敏感品系低;羧酸酯酶活性与敏感品系无明显差异。     

取食不同寄主植物的甜菜夜蛾对药剂敏感性的变化

初步探讨了取食不同寄主植物的甜菜夜蛾对药剂敏感性及其体内酶活性的变化。结果表明：取食不同寄主植物的甜菜夜蛾对灭多威、溴氰菊酯的敏感性变化较大，最大LD50之比分别达17．3和44．9倍，而对辛硫磷的敏感性变化不大，最大LD50之比为1．9倍；取食不同寄主植物的甜菜夜蛾体内谷胱甘肽-S-转移酶的活性差异不显著，而羧酸酯酶活性、多功能氧化酶脱甲基作用差异显著，其中羧酸酯酶的活性变化与对灭多威敏感性的变化相一致。因此，羧酸酯酶可能是寄主植物影响甜菜夜蛾对灭多威敏感性变化的因素之一。     

多功能氧化酶系与甜菜夜蛾对氯氟氰菊酯抗药性的关系

比较了甜菜夜蛾Spodoptera exigua (Hübner)抗氯氟氰菊酯品系和敏感品系5龄幼虫中肠、脂肪体及体壁微粒体细胞色素P450的含量,结果表明,抗性和敏感品系不同组织细胞色素P450的含量均为中肠＞脂肪体＞体壁,抗性品系中肠、脂肪体及体壁细胞色素P450的含量分别是敏感品系的1.78、1.54及1.37倍。中肠微粒体甲氧试卤灵-O-脱甲基酶、乙氧试卤灵-O-脱乙基酶、乙氧香豆素-O-脱乙基酶、芳香基羟基化酶及艾氏剂环氧化酶的活性测定结果表明,抗性品系中肠5种酶的活性分别比敏感品系的酶活性提高1.33、1.73、1.40、1.51及1.30倍,说明甜菜夜蛾对氯氟氰菊酯的抗药性与微粒体多功能氧化酶活性的提高密切相关。     

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。     

取食不同寄主植物的斜纹夜蛾对化学杀虫剂敏感性的变化

取食不同寄主植物的斜纹夜蛾３龄幼虫对溴氰菊酯（ｄｅｌｔａｍｅｔｈｒｉｎ）、氯氰菊酯（ｃｙｐｅｒｍｅｔｈｒｉｎ）和敌百虫（ｔｒｉｃｈｌｏｒｆｏｎ）的敏感性发生了变化，其中以取食甘薯的斜纹夜蛾３龄幼虫最为敏感，其次是木薯、甘蓝、萝卜、蓖麻和菜心；取食白菜的最不敏感。     

人工饲料对黄带犀猎蝽生长发育、繁殖及捕食效能的影响

为评价人工饲料饲养对广泛分布于我国南方的黄带犀猎蝽Sycanus bifidus的生长发育、繁殖和控害能力的影响,在室内条件下测定取食不同胶囊人工饲料的黄带犀猎蝽若虫生长发育和繁殖的相关生物学参数,并比较取食斜纹夜蛾Spodoptera litura胶囊人工饲料和活虫饲料的黄带犀猎蝽3~5龄若虫对斜纹夜蛾3龄幼虫的捕食功能反应和搜寻效应。结果表明,以斜纹夜蛾胶囊人工饲料饲喂的黄带犀猎蝽若虫可完成完整世代并继续繁殖,其成虫获得率为15.0%,若虫总发育历期为107.6 d,雌、雄成虫体重分别为113.8 mg和84.7 mg,产卵前期为31.3 d,单雌产卵量为117.3粒,卵期为24.5 d,卵孵化率为90.8%;而取食胶囊人工饲料内容物为黄粉虫Tenebrio molitor匀浆液和混合匀浆液的黄带犀猎蝽若虫不能完成完整世代。取食斜纹夜蛾胶囊人工饲料和活虫饲料的黄带犀猎蝽3~5龄若虫的捕食功能反应均符合Holling II型和Holling III型功能反应模型,使用斜纹夜蛾胶囊人工饲料饲养的黄带犀猎蝽对猎物的捕食效果和搜寻效应未受到明显影响,表明该人工饲料可用于黄带犀猎蝽的规模化扩繁。     

斜纹夜蛾取食为害对大豆农艺性状和土壤酶活性的影响

为研究斜纹夜蛾取食为害对大豆农艺性状和土壤酶活性的影响, 随机选取了5个大豆品种, ‘中黄13’‘中黄37’‘中黄39’‘石豆7号’和‘菏豆15’, 测试不同虫口密度的斜纹夜蛾取食为害后大豆的产量和蛋白质?脂肪含量及土壤酶活性?结果表明:大豆品种?斜纹夜蛾虫口密度以及两者间的互作对大豆的单株结荚数?单株荚粒数?单株产量?蛋白质含量?脂肪含量以及土壤脲酶和磷酸酶活性均有显著影响, 但对根际土壤酸碱度没有显著影响?随着虫口密度的增加, 大豆的结荚数?荚粒数?单株产量?蛋白质含量?脂肪含量减少, 土壤的脲酶活性和磷酸酶活性降低?综上所述, 斜纹夜蛾取食为害对大豆的产量和营养物质含量产生了不利影响, 对土壤酶活性产生了消极作用?     

Resistance to insecticides in winter- and summer-forms of pear psylla,Psylla pyricola

Summer-form pear psylla, Psylla pyricola Foerster, from sprayed pear were resistant to endosulfan (2·4-fold), methiocarb (2·5-fold), ethylan (5·8-fold), azinphos-methyl (7·7-fold), and fenvalerate (40·1-fold). Esterase (3·8-fold), glutathione transferase (1·8-fold), and cytochrome P-450 monooxygenase (1·6-fold) detoxification enzyme activity was higher in resistant than in susceptible summer forms. Synergism by piperonyl butoxide and S,S,S-tributylphosphorotrithioate (DEF) was added evidence for cytochrome P-450 monooxygenases and esterases as resistance mechanisms. Reduced penetration may also have contributed to resistance, as indicated by a 1·6-fold slower penetration of azinphos-methyl in resistant than susceptible summer-forms. Similar differences in insecticide toxicity and esterase and glutathione transferase activities were observed between winter-forms of resistant and susceptible pear psylla. Winter-forms of P. pyricola were up to three times more tolerant to insecticides than summer-forms. Higher cytochrome P-450 monooxygenase activity (1·7-fold) and slower azinphosmethyl penetration (2·1-fold) in winter-forms may have contributed to their greater insecticide tolerance; however, sequestration may also have been involved.     

The role of cytochrome P450 monooxygenase in the different responses to fenoxaprop-P-ethyl in annual bluegrass (Poa annua L.) and short awned foxtail (Alopecurus aequalis Sobol.)

Herbicide resistance or tolerance in weeds mediated by cytochrome P450 monooxygenase is a considerable problem. However, cytochrome P450 mediated resistance or tolerance in weeds was less studied. Thus, in this work, the role of the cytochrome P450 monooxygenase in the different responses of Poa annua and Alopecurus aequalis to fenoxaprop-P-ethyl was studied. We found that the effect of fenoxaprop-P-ethyl could be synergized by piperonyl butoxide (PBO) in P. annua, but not by malathion. After being treated with fenoxaprop-P-ethyl (containing mefenpyr-diethyl), the contents of cytochrome P450 and cytochrome b₅ in P. annua increased significantly compared to plants treated with mefenpyr-diethyl only or untreated plants. However, the increase was less in A. aequalis, which was susceptible to fenoxaprop-P-ethyl. The activities of ρ-nitroanisole O-demethylase (PNOD), ethoxyresorufin O-deethylase (EROD), ethoxycoumarin oxidase (ECOD) and NADPH-dependent cytochrome P450 reductase mediated by cytochrome P450 monooxygenase increased in P. annua after treatment with fenoxaprop-P-ethyl, especially the activities of ECOD and cytochrome P450 reductase. Besides this, cytochrome P450 monooxygenase activity toward fenoxaprop-P-ethyl in P. annua increased significantly compared to untreated or treated with mefenpyr-diethyl plants and treated or untreated A. aequalis. Cytochrome P450 monooxygenase may play an important role in the different responses to fenoxaprop-P-ethyl in P. annua and A. aequalis.     

大豆品种对斜纹夜蛾的抗性机制初探

在肉室人工接虫条件下研究大豆３个高抗品种吴江青豆３、通山薄皮黄豆甲、赶泰－２－和３个高感品种皖８２－１７８、山东大豆及Ｍｏｒｓｏｙ对斜纹夜蛾的抗性表现。结果表明,抗感材料间以叶面积损失率为指标的抗虫性差异极为,而对斜纹夜蛾产卵排趋性差异不显著。     

Co-expression of a NADPH:P450 reductase enhances CYP71A10-dependent phenylurea metabolism in tobacco

A soybean cytochrome P450 monooxygenase, designated CYP71A10, catalyzes the oxidative N-demethylation or ring methyl hydroxylation of a variety of phenylurea herbicides. The ectopic expression of CYP71A10 in tobacco was previously shown to be an effective means of enhancing whole plant tolerance to the compounds linuron and chlortoluron. Because P450 enzymes require ancillary proteins to catalyze the transfer of electrons from NADPH to the functional heme group of the P450, it is possible that the endogenous levels of these companion proteins may be insufficient to support the optimal activation of a highly expressed recombinant P450. In the present report, we have generated transgenic tobacco that simultaneously express CYP71A10 and a soybean P450 reductase. Transformed plants that express both CYP71A10 and the P450 reductase demonstrated 20-23% higher metabolic activity against phenylurea herbicides than control plants expressing CYP71A10 alone. These results suggest that herbicide tolerance strategies based on the expression of P450 genes may require concomitant expression of a cognate electron transport partner to fully exploit the herbicide metabolic capacity of the P450.     

棉铃虫雌成虫对16种植物的产卵偏好性及幼虫取食后的生存表现

为研究棉铃虫成虫寄主偏好性和幼虫取食后生存表现之间的关系,明确两者在决定寄主过程中的作用,我们测定了交配后的棉铃虫雌成虫对16种植物(棉花、玉米、大豆、花生、番茄、辣椒、茄子、烟草、黄瓜、胡萝卜、西芹、杨树、月季、菠菜、包菜、大葱)的产卵偏好性和幼虫取食这些植物嫩叶后的存活和发育情况。棉铃虫雌成虫选择在烟草、茄子、棉花、胡萝卜、番茄、辣椒、玉米、西芹、大豆和花生等10种植物上产卵,但只有取食西芹、烟草、番茄、大豆、棉花和胡萝卜等6种植物叶片的初孵幼虫能够完成幼虫阶段的生长发育、化蛹并羽化,且取食植物嫩叶的幼虫的存活率、发育历期、蛹期和蛹重均差于取食人工饲料的幼虫。以上结果表明,棉铃虫雌成虫产卵寄主与其幼虫取食寄主之间存在并非一一对应的关系,幼虫的生存表现限定了棉铃虫的寄主范围。     

Mechanisms for multiple resistances in field populations of common cutworm, Spodoptera litura (Fabricius) in China

The mechanisms for multiple resistances had been studied with two field resistant strains and the selected susceptible and resistant strains of Spodoptera litura (Fabricius). Bioassay revealed that the two field strains were both with high resistance to pyrethroids (RR: 63-530), low to medium resistance to organophosphates and carbamates, AChE targeted insecticides (RR: 5.7-26), and no resistance to fipronil (RR: 2.0-2.2). Selection with deltamethrin in laboratory could obviously enhance the resistance of this pest to both pyrethroids and AChE targeted insecticides. Synergism test, enzyme analysis and target comparison proved that the pyrethroid resistance in this pest associated only with the enhanced activity of cytochrome P450 monooxygenase (MFO) and esterase. However the resistance to the AChE targeted insecticides depended on the target insensitivity and also the enhanced activity of MFO and esterase. Thus, the cross-resistance between pyrethroids and the AChE targeted insecticides was thought to be resulted from the enhanced activity of MFO and esterase.     

抗阿维菌素小菜蛾的细胞色素P450酶系研究

通过对不同发育时期敏感和抗阿维菌素小菜蛾品系细胞色素P450含量的测定,以及使用不同模式底物对P450单加氧酶活性的比较研究发现:除成虫期外,不同发育时期抗性品系小菜蛾中P450和细胞色素b5的含量都高于敏感品系;抗性品系还原型辅酶Ⅱ(NADPH)-细胞色素P450还原酶活性是敏感品系的1.97倍;同时发现抗性品系中甲氧试卤灵-O-脱甲基酶(MROD)、乙氧试卤灵-O-脱乙基酶(EROD)、乙氧基香豆素-O-脱乙基酶(ECOD)以及对硝基苯甲醚-O-脱甲基酶(PNOD)的活性均明显高于敏感品系,分别为敏感品系的9.41、4.15、1.67和2.94倍。研究结果表明,细胞色素P450含量和单加氧酶活性的增高是小菜蛾对阿维菌素产生抗性的一个重要机制。     

Delayed cuticular penetration and enhanced metabolism of deltamethrin in pyrethroid-resistant strains of Helicoverpa armigera from China and Pakistan

The penetration and metabolism of [14C]deltamethrin was studied in susceptible and resistant Chinese and Pakistani strains of Helicoverpa armigera (Hübner), which were resistant to deltamethrin by 330- and 670-fold, respectively. The penetration of deltamethrin into resistant individuals was significantly slower than into susceptible individuals over a 24-h period. The time taken for 50% penetration of the applied deltamethrin was 1 h for the susceptible strain and 6 h for both of the resistant strains. The internal radioactivity was reduced by the larvae of resistant strains much faster than by the susceptible larvae. After 48 h, 40% of the penetrated deltamethrin was still inside the larvae of the susceptible strain, in comparison with 1.5-5% in the Pakistani strain and 8-14% in the Chinese strain. Both of the resistant strains produced methanol-soluble and water-soluble metabolites, but the susceptible strain produced methanol-soluble metabolites only. By 12, 24 and 48 h after dosing, the amount of methanol-soluble metabolites excreted by the resistant strains was almost double that of the susceptible strain. Both of the resistant strains also excreted 5-7% of the penetrated dose as a water-soluble metabolite after 48 h. In comparison with the Chinese strain, the Pakistani strain exhibited slower penetration, lower internal content and faster excretion of deltamethrin, which correlated with the higher resistance of the Pakistani strain. These findings show that the resistant Pakistani and Chinese strains of H. armigera possess mechanisms of reduced cuticular penetration and enhanced metabolism of deltamethrin and perhaps other pyrethroids.     

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.