Acetylcholinesterase point mutations in European strains of Tetranychus urticae (Acari: Tetranychidae) resistant to organophosphates

BACKROUND: In Tetranychus urticae Koch, acetylcholinesterase insensitivity is often involved in organophosphate (OP) and carbamate (CARB) resistance. By combining toxicological, biochemical and molecular data from three reference laboratory and three OP selected strains (OP strains), the AChE1 mutations associated with resistance in T. urticae were characterised. RESULTS: The resistance ratios of the OP strains varied from 9 to 43 for pirimiphos‐methyl, from 78 to 586 for chlorpyrifos, from 8 to 333 for methomyl and from 137 to 4164 for dimethoate. The insecticide concentration needed to inhibit 50% of the AChE1 activity was, in the OP strains, at least 2.7, 55, 58 and 31 times higher for the OP pirimiphos‐methyl, chlorpyrifos oxon, paraoxon and omethoate respectively, and 87 times higher for the CARB carbaryl. By comparing the AChE1 sequence, four amino acid substitutions were detected in the OP strains: (1) F331W (Torpedo numbering) in all the three OP strains; (2) T280A found in the three OP strains but not in all clones; (3) G328A, found in two OP strains; (4) A201S found in only one OP strain. CONCLUSIONS: Four AChE1 mutations were found in resistant strains of T. urticae, and three of them, F331W, G328A and A201S, are possibly involved in resistance to OP and CARB insecticides. Among them, F331W is probably the most important and the most common in T. urticae. It can be easily detected by the diagnostic PCR‐RLFP assay developed in this study. Copyright © 2009 Society of Chemical Industry     

地椒提取物杀螨活性研究

从地椒中提取挥发油及乙醇提取物,研究并比较了地椒乙醇提取物和挥发油对棉红蜘蛛的触杀活性。结果显示地椒乙醇提取物对棉红蜘蛛显示出很强的触杀作用,其毒力回归方程为Y=4.2967+1.1792x,LD50为3.95 mg/mL;地椒挥发油对棉红蜘蛛也具有显著的触杀活性,触杀作用的毒力回归方程为Y=4.6691+1.4427,LD50为1.70 mg/mL,低于乙醇提取物LD50,显示其杀螨活性较乙醇提取物更高。此外,杀卵活性试验也显示地椒挥发油的杀卵作用要高于地椒乙醇提取物,其挥发油和乙醇提取物的杀卵LC50分别为2.241和5.714 mg/mL。     

18g／L阿维菌素乳油防治棉花红蜘蛛试验

进行田间试验以明确18g/L阿维菌素乳油防治棉花红蜘蛛的效果。结果表明：18g,L阿维菌素乳油（8．1～10．8）g／hm^2能够很好的控制棉花红蜘蛛,且防治效果高于对照,对作物安全。     

截形叶螨取食对马铃薯酶活性的影响

为明确截形叶螨(Tetranychus truncatus)取食刺激对不同耐旱型马铃薯(Solanum tuberosum)品种酶活性的影响,选择2个马铃薯品种（‘荷兰15号’,早熟,干旱敏感型品种;‘陇薯10号’,晚熟,耐旱型品种）,在人工接螨后,测定7天内马铃薯叶片5种防御酶(CAT、POD、PPO、PAL、LOX)活性的变化。CAT、POD、PPO、PAL、LOX等5种酶活性在受螨害刺激后均表现出不同程度的升高,在时序上都表现为先升高后下降的趋势,且在3天后显著高于CK（未接螨的健康植株）,5种酶均参与了植物机体对害螨的防御反应,以促进受害植株体内自由氧的消减,通过木质素、花青素、生物碱等苯丙素类化合物、氢过氧化物等含量的提高来增强机体防御能力;在螨害7天后,5种酶活性都显著下降,且显著低于CK,说明叶螨在长时间危害后可能导致植株机体紊乱,植株不能很好地持续维持高效的对螨害的防御响应。2个马铃薯品种在叶螨为害后同一时间内,从CAT、PPO和PAL活性变化角度分析,干旱敏感型马铃薯品种表现出较高的抗螨性;被叶螨取食为害后耐旱型马铃薯品种POD活性较高,但干旱敏感型马铃薯品种POD活性增长幅度更大;2个品种在LOX活性上无显著差异。总体分析可得,干旱敏感型马铃薯比耐旱型品种有更高的抗螨性。研究结果可为马铃薯叶螨的科学防治和品种选育提供理论依据。     

朱砂叶螨乙酰胆碱酯酶的表达纯化及其酶活测定条件优化

乙酰胆碱酯酶(Acetylcholinesterase,AChE)是生物体内保证神经信号正常传递的关键酶,是有机磷和氨基甲酸酯类农药的作用靶标。现研究拟采用发酵罐培养含pET-30a/ace的表达菌株E.coli BL21(DE3),大量表达纯化螨AChE,利用单因素试验和正交试验优化AChE活性的测定条件。结果表明:发酵培养的重组菌株大量表达约为68kD的朱砂叶螨AChE蛋白。底物浓度、温度、pH和酶浓度等各测定因素对朱砂叶螨乙酰胆碱酯酶活性存在显著影响。确定朱砂叶螨乙酰胆碱酯酶活性测定的最佳反应条件为:底物浓度1mmol/L,pH值7,温度35℃,酶浓度0.5mg/mL。通过体外发酵培养纯化获得了大量的朱砂叶螨AChE重组蛋白,确定了螨AChE活性测定的最佳条件,为进一步研究其酶学性质以及AChE抑制剂的体外筛选奠定基础。     

Olfactory response of the ladybird beetle <Emphasis Type="Italic">Stethorus gilvifrons</Emphasis> to two preys and herbivore-induced plant volatiles

The spider mites Tetranychus urticae Koch and Panonychus ulmi (Koch) (Tetranychidae) cause severe economic losses to vegetable farms and deciduous fruit orchards in Turkey. One of their predators, the ladybird beetle Stethorus gilvifrons (Muls.) (Col., Coccinellidae), aggregates on mite-infested patches of plants. The present study assessed whether there is a role for herbivore-induced plant volatiles (HIPVs) and/or odors emitted directly from these two mite species in the aggregative response of ladybird beetles. The olfactory responses of the predator females to volatiles from T. urticae- and/or P. ulmi-infested sweet pepper (four cultivars, viz. ‘Demre’, ‘Yalova Carliston’, ‘Kandil Dolma’ and ‘Yag Biberi’), kidney bean (cv. ‘Barbunya’) and apple (M9 rootstock) were investigated using a two-choice olfactometer. Our results showed that HIPVs emitted from both T. urticae- and P. ulmi-infested plants significantly attracted S. gilvifrons adults for all plants except the sweet pepper cv. Yag Biberi. In addition, it was found that volatiles from apple plants infested by T. urticae and, especially, P. ulmi are more attractive for S. gilvifrons females than those emitted by other infested plants. The results also suggest that the odors of T. urticae adults and their products might influence the attraction of S. gilvifrons females.     

假蒟提取物对皮氏叶螨的生物活性测定

通过索氏抽提和乙醇浸提法提取假蒟(Rper.saormentosum Roxb.)杀虫活性成分,即获得乙醇浸提物、乙醇索氏提取物和石油醚索氏提取物,研究其对皮氏叶螨(Tetranychus pieroei McGregor)各龄虫的触杀及对成螨驱避、产卵抑制等方面的生物活性.结果表明:乙醇浸提物和石油醚索氏提取物对皮氏叶螨卵的毒杀效果均较好,用20mg/g的浓度处理后,杀卵效果均在95%以上:对若螨的生物活性以乙醇索氏提取物最好,当浓度为15mg/g时,校正死亡率达95.19%,明显强于石油醚索氏提取物;乙醇浸提物、乙醇索氏提取物和石油醚索氏提取物对成螨的LC50分别为3.2141,6.093 1,3.518 8 mg/g,LC95分别为8.5750,18.700 7,9.311 4 mg/g;用质量分数0.4%乙醇浸提物和石油醚索氏提取液处理的皮氏叶螨成螨寿命明显短于乙醇索氏提取物和对照,产卵抑制指数以石油醚索氏提取物最高,为0.1465,乙醇索氏提取物最低,为0.0825;3种提取物对皮氏叶螨均有良好的驱避作用.     

The discovery of HNPC‐A3066: a novel strobilurin acaricide

BACKGROUND: Although more than ten strobilurin analogues have been commercialized since 1996 as fungicides, only one was available as an acaricide as of 2003. To search for novel strobilurin analogues with unique biological activities, a synthetic screening programme was carried out. RESULTS: Syntheses of compounds analogous to the commercialized fungicide metominostrobin and the acaricide fluacrypyrim led to the discovery of a lead compound, (E)‐2‐{2‐[[3,5‐bis(trifluoromethyl)phenoxy]methyl]phenyl}‐2‐(methoxyimino)‐N‐methylacetamide (3b), that showed moderate acaricidal activity against Tetranychus urticae Koch. Compound 3b has a 3,5‐(CF₃)₂‐phenoxymethyl group instead of the unsubstituted phenoxy substituent in metominostrobin. Optimization of compound 3b was achieved by introducing an oxime ether bridge along with an alkylthio(alkyl) branch in place of the oxymethylene chain between two aromatic moieties, as well as by replacing the methoxyiminoacetamide group with a methoxyacrylate structure, leading to (E)‐ methyl 2‐{2‐[[[(Z)[1‐(3,5‐bis(trifluoromethyl)phenyl)‐2‐methylthioethylidene]amino]oxy] methyl]phenyl}‐3‐methoxyacrylate (6c) and (E)‐ methyl 2‐{2‐[[[(Z)[1‐(3,5‐bis(trifluoromethyl)phenyl)‐1‐methylthiomethylidene]amino]oxy]methyl]phenyl}‐3‐methoxyacrylate (9a, HNPC‐A3066). CONCLUSION: The above two compounds (6c, 9a) were shown to exhibit potent acaricidal and fungicidal activity. Compound 9a (HNPC‐A3066) also exhibits larvicidal and ovicidal activities against various acarids. The acaricidal potency is comparable with those of commercial acaricides such as fluacrypyrim, tebufenpyrad and chlorfenapyr. Copyright © 2008 Society of Chemical Industry     

土耳其斯坦叶螨危害对棉花叶片叶绿素、可溶性蛋白质、可溶性糖影响的研究

棉花受土耳其斯坦叶螨危害0～8d内,叶绿素、可溶性糖、蛋白质含量随危害程度的加深均有不同程度的变化.在试验期内,两处理叶片叶绿素a、叶绿素b及叶绿素总量均随危害时间的延长而下降,不同危害时间之间下降的幅度差异显著(P<0.05),另外,相同危害时间下各处理叶片叶绿素a、叶绿素b及叶绿素总量下降的幅度也随螨口密度的增大而增加,不同螨口密度处理叶片之间的叶绿素含量差异显著(P<0.05).在试验期内,受害叶片可溶性糖及蛋白质含量随危害时间的延长变化不稳定.     

释放中华微刺盲蝽防治茄子害虫的研究

采用3．00、1．50和0．75头／m^2 3种释放密度,探讨了释放中华微刺盲蝽对大棚内茄子上节瓜蓟马、烟粉虱和朱砂叶螨等3种害虫的控制作用,结果表明中华微刺盲蝽在释放区能形成种群;在0．75头／m^2的释放密度下,该盲蝽已能很好地控制茄子上的3种害虫,释放后20～30d控制效果达100％．