二斑叶螨对阿维菌素抗性的CAPS标记

为了实现二斑叶螨Tetranychus urticae Koch对阿维菌素抗药性的快速检测,本研究建立了一种叶螨对阿维菌素抗药性的快速分子检测技术,即酶切扩增多态性序列(cleaved amplified polymorphic sequences,CAPS)标记技术,检测二斑叶螨谷氨酸门控氯离子通道(Glu Cl3)基因片段上G326E的突变频率,并采用琼脂浸叶法测定田间5个种群对阿维菌素的抗药性。结果表明,北京密云、顺义、昌平、浙江宁波和海南吉阳5个种群对阿维菌素的抗性倍数分别达1 155.63、1 317.10、844.19、314.95和1 799.69倍,均为极高抗性水平;CAPS检测发现我国二斑叶螨抗性基因存在G326E位点突变,但在供试的5个不同种群中突变频率变异很大,其中北京昌平种群G326E突变频率高达90.00%,密云和顺义种群均为26.67%,浙江宁波种群为5.00%,而海南吉阳种群不存在该位点的突变。表明该CAPS标记还需与其它抗性机制或检测技术结合来实现叶螨对阿维菌素抗性的早期检测和预警。     

蒜和葱上二斑叶螨的发生为害及其抗药性监测

二斑叶螨寄主植物范围极广, 但其对百合科蔬菜造成危害的研究报道极少?本研究采集了来自北京海淀百合科蔬菜蒜和葱上的2个二斑叶螨种群, 生物测定结果显示, 2个种群对阿维菌素呈现出极高水平抗性, 对哒螨灵为中等以上抗性(LC50>5 000 mg/L), 对新型药剂联苯肼酯?虫螨腈和腈吡螨酯也表现出中等到高水平抗性?抗性基因突变频率检测发现, 与阿维菌素抗性相关基因G314D和G326E位点以及和哒螨灵抗性相关基因H92R位点的突变频率均高达100%, 为纯合抗性种群?因此, 本研究所测试的2个二斑叶螨种群对阿维菌素和哒螨灵单剂的敏感性极低, 需慎重选用这2种药剂, 而3种新型杀螨剂联苯肼酯?虫螨腈和腈吡螨酯在田间需轮换使用?     

几种杀螨剂对二斑叶螨和朱砂叶螨的毒力测定

作者选用三氯杀螨醇、克螨特和螨克2种杀螨剂对朱砂叶螨(儋州、岳阳和射阳种群)和二斑叶螨作了毒力测定,明确3种杀螨剂对二斑叶螨的LC₅₀明显高于对朱砂叶螨,差异显著,已不能用于二斑叶螨的防治。螨克也已不适用于岳阳地区朱砂叶螨的防治。三氯杀螨醇和克螨特对朱砂叶螨儋州、岳阳和射阳种群的LC₅₀分别为237.79和804.64ppm、129.07和830.98ppm、112.03和732.32ppm,尚可在一定范围内用于对上述种群的防治,但应加强监测,注意与其他杀螨剂轮用,以延缓其使用期限。     

抗梅岭霉素和阿维菌素二斑叶螨种群生命力和繁殖力的研究

采用营养液培养的棉苗饲养二斑叶螨Tetranychus urticae Koch,用梅岭霉素、阿维菌素对其逐代进行抗性选育,至第9代,测定了两个抗性种群的抗性水平、生命力和繁殖力。结果表明:梅岭霉素抗性种群的抗性达7.9倍,其雌成螨寿命、产卵量、卵的孵化率和后代若螨成活率分别比对照增加54.08%、39.93%、6.57%和-15.44%。阿维菌素抗性种群的抗性达5.8倍,其雌成螨寿命、产卵量、卵的孵化率和后代若螨成活率分别比对照增加15.88%、10.87%、5.86%和-9.32%。由此发现梅岭霉素和阿维菌素对二斑叶螨的生长发育和繁殖有一定有利作用,抗性风险性较大,应引起重视。     

加州新小绥螨对朱砂叶螨的控制作用

为明确加州新小绥螨Neoseiulus californicus（McGregor）国内种群对朱砂叶螨Tetranychus cinnabarinus（Boisduval）的控制潜力,应用功能反应、数值反应及实验种群生命表参数评价了该捕食螨对朱砂叶螨的控制能力。加州新小绥螨对朱砂叶螨的功能反应能很好地拟合Holling Ⅱ方程。在25℃,加州新小绥螨雌成螨对朱砂叶螨卵、幼螨、若螨和成螨的功能反应参数a/T_h值分别为42.4195、81.6275、54.3044和17.9399,对朱砂叶螨幼螨、若螨和卵的控制力高于成螨。在19~31℃,加州新小绥螨对朱砂叶螨雌成螨日均捕食量和对猎物的控制能力a/T_h随温度升高而增大,在31℃时达最大值,分别为10.80头和29.5364。在28℃时,加州新小绥螨的净增殖率和内禀增长率分别为26.1522和0.2213。表明加州新小绥螨国内种群表现出重要的生物防治潜能。     

二斑叶螨对螺螨酯抗药性及对18种杀螨剂交互抗性

采自甘肃兰州兴隆山公园的二斑叶螨(Tetranychus urticae Koch),用雌雄单系培养敏感品系(S),用螺螨酯处理二斑叶螨种群培养抗性品系(SP R),用室内生测法对二斑叶螨S和SP R品系进行室内毒力测定。结果表明,二斑叶螨对螺螨酯抗性发展初期较慢,中期稳定,后期较快,选育至26代抗性指数(RI)达58.83。SP R对甲氰菊酯、氯氰菊酯有一定的交互抗性,RI分别为11.54和10.03;对苯丁锡、四螨嗪、苦皮藤生物碱、阿维菌素、氯氟氰菊酯、哒螨&#8226;四螨嗪、哒&#8226;水胺硫磷、三唑锡、三氯杀螨醇、哒螨灵、氧化乐果无交互抗性(1＜RI<5.00);对浏阳霉素、毒死蜱、噻螨酮、柴油、哒螨灵、唑螨酯可能存在负交互抗性(RI<1)。     

15种常用杀虫剂对二斑叶螨的室内药效对比试验

本文采用FAO推荐的玻片浸渍法,选用15种常用杀虫剂的最大登记剂量对二斑叶螨敏感型种群和抗性种群进行了药效对比试验研究。结果表明:施药后观察,哒螨·矿物油、阿维·哒螨灵、阿维菌素、阿维·灭幼脲、阿维·毒死蜱、啶虫脒对敏感型二斑叶螨的速效性防效较好;而炔螨特、桉油精和苦参碱对抗性二斑叶螨的防效较强。     

我国部分地区田间小菜蛾种群对8种常用杀虫剂的抗性检测

采用浸叶法测定了来自5个地区的田间小菜蛾种群对8种杀虫剂的抗性水平.结果 表明,5个地区小菜蛾种群对氯虫苯甲酰胺均产生了中等水平以上的抗性,江苏无锡和广东增城种群的抗性达1 428.16倍和6 642.12倍,为极高抗性水平;对丁醚脲均处于敏感或敏感性降低水平.5个地区的小菜蛾种群对甲氨基阿维菌素、虫螨腈、S-茚虫威、...     

二斑叶螨对甲氰菊酯和螺螨酯的抗性选育及增效剂的增效作用

［目的］ 探索二斑叶螨对甲氰菊酯和螺螨酯种群的抗性机理和抗性治理途径。 ［方法］ 采用室内生物测定法,以采自兰州兴隆山自然保护区的二斑叶螨为敏感种群（S）,在室内用甲氰菊酯和螺螨酯分别对二斑叶螨进行抗性选育;用增效醚(PBO)、顺丁烯二酸二乙酯(DEM)、磷酸三甲苯酯(TPP)、有机硅、噻酮进行增效作用研究。［结果］二斑叶螨经室内甲氰菊酯和螺螨酯抗性选育45代和30代后,抗性倍数分别达到314.50倍和77.92倍。TPP、PBO、DEM 3种增效剂对二斑叶螨抗甲氰菊酯种群的抗敏增效比分别为12.15、7.78和3.09,推测其抗性机理涉及的主要解毒酶是羧酸酯酶和多功能氧化酶; DEM对二斑叶螨抗螺螨酯种群的抗敏增效比大于TPP和PBO,分别为4.87、3.67和1.91,二斑叶螨对螺螨酯产生抗性机理可能与谷胱甘肽转移酶和羧酸酯酶活性增强有关。有机硅和噻酮对二斑叶螨抗甲氰菊酯和螺螨酯种群的抗敏增效指数比分别为1.38、1.42和1.18、0.92,说明二斑叶螨的抗性与表皮通透性改变关系不密切。 ［结论］ 上述结果可以为二斑叶螨的抗性治理提供依据。     

温度对阿维菌素(Avermectin)杀螨活性的影响

研究了不同恒温条件下阿维菌素(Avermectin)对朱砂叶螨[Tetranychus cinnabarinus(Boisduval)]和二斑叶螨(T.urticae Koch)的致死作用,拟合了阿维菌素对两种叶螨的毒力与温度之间的关系。结果表明:阿维菌素分别在16、21、26、31℃和36℃的恒温条件下,对两种叶螨的毒力与温度呈正相关关系,即温度越高毒力越强;朱砂叶螨对阿维菌素的敏感性高于二斑叶螨。     

Chlorfenapyr resistance in the spider mite Tetranychus urticae: stability, cross-resistance and monitoring of resistance

The two-spotted spider mite, Tetranychus urticae Koch, is a key pest of many agricultural crops. Studies of stability of resistance, cross-resistance relationships and monitoring of chlorfenapyr resistance were carried out with T. urticae to provide basic information necessary to define effective acaricide resistance management strategies for this pest. Chlorfenapyr resistance was shown to be stable in the absence of selection pressure under laboratory conditions. The activities of seven different acaricides against chlorfenapyr-resistant and -susceptible strains of T. urticae were evaluated. The results indicated possible positive cross-resistance between chlorfenapyr and the acaricides abamectin, propargite and etoxazole. No cross-resistance was detected for the acaricides milbemectin, fenpyroximate and diafenthiuron. A possible negatively correlated cross-resistance was observed between chlorfenapyr and spiromesifen. The evaluation of 21 T. urticae populations from several crops in the States of São Paulo, Mato Grosso, Goiás, and Bahia, in Brazil, indicated that the susceptibility of mites to chlorfenapyr was variable, with percentages of resistant mites ranging from 0.0 to 86.5%. The highest resistance frequencies were observed in ornamental plants in the State of São Paulo. Some populations from cotton and papaya also presented high frequencies of chlorfenapyr resistance. This is the first report on chlorfenapyr resistance in T. urticae on cotton and papaya in Brazil. Strategies for the management of acaricide resistance are discussed.     

北京地区板栗主要病虫害种类初步调查

2006-2007年,在北京周边的怀柔、密云、昌平、平谷和延庆5个区县共36个调查点对板栗病虫害种类及危害状况进行了实地调查,初步查清了北京地区板栗生产中主要发生的病虫害种类,在此基础上提出板栗疫病、针叶小爪螨和桃蛀螟是当前板栗生产中病虫害防治的主要对象。     

北京、山东和湖南地区烟粉虱抗药性及CYP4v2和CYP6CX1 mRNA水平表达量分析

于2011年采集北京、山东和湖南三地的烟粉虱,进行B、Q隐种鉴定,并测定4种杀虫剂的抗药性,同时通过荧光定量PCR分析CYP4v2和CYP6CX1两个基因的mRNA水平的表达量。结果表明,北京、湖南和长沙烟粉虱均为Q隐种。抗药性监测表明,北京和湖南种群对阿维菌素敏感,山东种群抗性水平较低,而对烟碱类药剂噻虫嗪出现不同程度的抗药性,其中湖南地区烟粉虱对噻虫嗪的抗药性达到49.08倍的高抗水平,北京和山东地区也达到中抗水平。另外,这3个地区的种群对毒死蜱和联苯菊酯抗性水平都较低。通过qRT-PCR分析三地的CYP4v2和CYP6CX1基因表达量,发现相对于敏感种群CYP4v2基因在北京、山东和湖南3个地理种群中分别过量表达3.85倍、19.57倍和10.78倍,而CYP6CX1基因在北京种群中过量表达20.55倍。结果提示田间烟粉虱的细胞色素P450基因CYP4v2和CYP6CX1过量表达可能会是烟粉虱抗药性的形成机制之一。     

Acetylcholinesterase point mutations putatively associated with monocrotophos resistance in the two-spotted spider mite

Molecular mechanisms of monocrotophos resistance in the two-spotted spider mite (TSSM), Tetranychus urticae Koch, were investigated. A monocrotophos-resistant strain (AD) showed ca. 3568- and 47.6-fold resistance compared to a susceptible strain (UD) and a moderately resistant strain (PyriF), respectively. No significant differences in detoxification enzyme activities, except for the cytochrome P450 monooxygenase activity, were found among the three strains. The sensitivity of acetylcholinesterase (AChE) to monocrotophos, however, was 90.6- and 41.9-fold less in AD strain compared to the UD and PyriF strains, respectively, indicating that AChE insensitivity mechanism plays a major role in monocrotophos resistance. When AChE gene (Tuace) sequences were compared, three point mutations (G228S, A391T and F439W) were identified in Tuace from the AD strain that likely contribute to the AChE insensitivity as predicted by structure analysis. Frequencies of the three mutations in field populations were predicted by quantitative sequencing (QS). Correlation analysis between the mutation frequency and actual resistance levels (LC₅₀) of nine field populations suggested that the G228S mutation plays a more crucial role in resistance (r² = 0.712) compared to the F439W mutation (r² = 0.419). When correlated together, however, the correlation coefficient was substantially enhanced (r² = 0.865), indicating that both the F439W and G228S mutations may work synergistically. The A391T mutation was homogeneously present in all field populations examined, suggesting that it may confer a basal level of resistance.     

Purification and partial characterization of a glutathione S-transferase from the two-spotted spider mite, Tetranychus urticae

Glutathione S-transferases (GSTs) are known to catalyze conjugations by facilitating the nucleophilic attack of the sulfhydryl group of endogenous reduced glutathione on electrophilic centers of a vast range of xenobiotic compounds, including insecticides and acaricides. Elevated levels of GSTs in the two-spotted spider mite, Tetranychus urticae Koch, have recently been associated with resistance to acaricides such as abamectin [Pestic. Biochem. Physiol. 72 (2002) 111]. GSTs from acaricide susceptible and resistant strains of T. urticae were purified by glutathione-agarose affinity chromatography and characterized by their Michaelis-Menten kinetics towards artificial substrates, i.e., 1-chloro-2,4-dinitrobenzene and monochlorobimane. The inhibitory potential of azocyclotin, dicumarol, and plumbagin was low (IC₅₀ values > 100 μM), whereas ethacrynic acid was much more effective, exhibiting an IC₅₀ value of 4.5 μM. GST activity is highest in 2-4-day-old female adults and dropped considerably with progressing age. Furthermore, molecular characteristics were determined for the first time of a GST from T. urticae, such as molecular weight (SDS-PAGE) and N-terminal amino acid sequencing (Edman degradation). Glutathione-agarose affinity purified GST from T. urticae strain WI has a molecular weight of 22.1 kDa. N-terminal amino acid sequencing revealed a homogeneity of ≈50% to insect GSTs closely related to insect class I GSTs (similar to mammalian Delta class GSTs).     

不同种群小菜蛾对4种杀虫药剂的抗性及抗性基因频率

用 FAO推荐的点滴法测定了日本、台湾、广州、东莞等地小菜蛾 (Plutella xy-lostella L .)的抗药性。测定结果表明 :日本、广州的小菜蛾对阿维菌素显示出抗药性 ,其抗性比率分别是 4 .85倍和 10 .2 7倍。杀虫药剂抗性基因型频率测定显示 ,广州、广东东莞和北京种群的最高 ,基因型频率分别为 4 1.2 5%、4 1.2 5%和 58.2 3%。研究验证了快速测定杀虫药剂抗性基因型频率的方法 ,并提出了根据不同种群小菜蛾抗性基因型频率所应采取的综合治理策略。     

First molecular genotyping of voltage gated sodium channel alleles in Culex quinquefasciatus populations in Malaysia

A nationwide investigation was performed to detect the presence of 1014 mutation(s) in voltage gated sodium channel (kdr) gene of Culex quinquefasciatus from 14 residential areas across 13 states and a federal territory in Malaysia. Molecular genotyping of kdr mutation was performed via a modified three tubes allele-specific-polymerase chain reaction (AS-PCR) and direct sequencing of kdr gene. Based on the results of AS-PCR, homozygous susceptible (SS) genotype was found in nine out of 14 populations with 38 individuals from a total sample size of 140. Heterozygous (RS) genotype was most predominant (99 individuals) and distributed across all study sites. Homozygous resistance (RR) genotype was detected in Perak (one individual) and Selangor (two individuals). The resistance kdr allele frequencies ranged from 0.1 to 0.55, with the highest being detected in Cx. quinquefasciatus population from Selangor. This study has documented the first field-evolved instance of 1014F mutation in Malaysian mosquitoes and the findings of this study could be utilized in the implementation of strategic measures in vector control programs in Malaysia.     

Frequency detection of pyrethroid resistance allele in Anopheles sinensis populations by real-time PCR amplification of specific allele (rtPASA)

To investigate the level of pyrethroid resistance in Anopheles sinensis Wiedemann 1828 (Diptera: Culicidae), a major malaria vector in Korea, we cloned and sequenced the IIS4-6 transmembrane segments of the sodium channel gene that encompass the most widely known kdr mutation sites. Sequence analysis revealed the presence of the major Leu-Phe mutation and a minor Leu-Cys mutation at the same position in permethrin-resistant field populations of An. sinensis. To establish a routine method for monitoring resistance, we developed a simple and accurate real-time PCR amplification of specific allele (rtPASA) protocol for the estimation of resistance allele frequencies on a population basis. The kdr allele frequency of a field population predicted by the rtPASA method (60.8%) agreed well with that determined by individual genotyping (61.7%), demonstrating the reliability and accuracy of rtPASA in predicting resistance allele frequency. Using the rtPASA method, the kdr allele frequencies in several field populations of An. sinensis were determined to range from 25.0 to 96.6%, suggestive of widespread pyrethroid resistance in Korea.