改进蝇类昆虫触角电位技术的尝试

本文通过对美洲斑潜蝇触角的扫描电镜观察及触角电位的测定 ,提出对蝇类昆虫触角电位测定技术的改进方法 :将触角芒切断 ,露出血淋巴 ,记录电极与触角芒的断面相接 ,参照电极通过头孔插入脑血腔形成回路。     

昆虫神经生物学研究技术:触角电位图记录

触角电位图记录是昆虫神经生物学中用于昆虫嗅觉、信息素生物测定研究的重要技术之一。该技术具有敏感性、选择性高及操作简单易行等优点。该文以荷兰Syntech公司的产品为例详细介绍了此技术原理、装置、操作及应用     

气相色谱与触角电位检测器联用技术及其应用

介绍了气相色谱与触角电位检测器联用技术的工作原理、优点及其在昆虫性信息素或其它信息物质鉴定方面的应用。根据作者应用此技术进行赤松毛虫和桑尺蛾性信息素研究的经验 ,重点介绍了气相色谱与触角电位联用技术在实践操作中应注意的问题     

棉铃虫对几种信息化合物的触角电位 (EAG)反应

通过棉铃虫成虫对 1 0种寄主植物挥发性物质、两种性信息素组分及两者的混合作用的触角电位反应 ( EAG) ,发现棉铃虫雌雄蛾对 1 0种挥发性物质的 EAG反应差异显著 ( P<0 .0 5 ) ,说明了 1 0种挥发物质对棉铃虫成虫的感应功能有所不同。 1 0种寄主植物挥发物质与性信息素主要组分混合后能引起雄蛾 EAG反应明显高于单独性信息素的反应 ,其中有 4种挥发性物质明显地增强棉铃虫对性信息素的反应 ( P<0 .0 5 ) ,即庚醛、1 -己醇、反 - 2 -己烯醇、顺 - 3-己醇 - 1 ,说明了以寄主植物挥发物质与昆虫性信息素混合作用来增强昆虫性信息素的应用效果。不同光温条件下饲养的雄蛾对寄主植物挥发性物质与性信息素相互作用的 EAG反应差异显著 ( P<0 .0 5 )。     

昆虫对宿主植物的嗅觉定向

在混虫辨别宿主植物所利用的各种信息中,植物的气味是一个很重要的因素昆虫利用触角上的嗅觉感受器来感觉植物气味。触角电位是昆虫的触角感受到植物气味后所产生的神经反应,而嗅觉编码则是嗅觉感受器将植物的气味信息传至神经中枢的脉冲系列。本文从行为生理学的角度介绍了昆虫利用植物气味寻找宿主植物的过程。     

黄秋葵挥发油对小菜蛾的触角电位反应及趋性研究

通过触角电位仪测定了小菜蛾Plutella xylostella(L.)对黄秋葵(Abelmoschus esculentus(L.)Moench Meth)挥发油的触角电位反应。结果表明,黄秋葵挥发油对小菜蛾成虫的触角电位反应强度与挥发油剂量成正比。四臂嗅觉仪实验进一步表明,黄秋葵挥发油对小菜蛾起到驱避作用。     

长足大竹象的触角超微结构和对竹笋挥发物的触角电位反应

长足大竹象Cyrtotrachelus buqueti Guerin-Meneville是当前竹林主要害虫, 专一地取食危害丛生竹笋梢。本研究应用电镜扫描、气质色谱联用(GC-MS)及触角电位(EAG)技术, 分析了长足大竹象成虫触角的超微结构、不同竹笋挥发物的成分及其与成虫的触角电位反应, 以期分析长足大竹象寡食竹笋的原因。结果表明长足大竹象成虫触角上感知竹笋挥发物的锥形感器数量最多、密度最大。气质色谱联用在竹笋挥发性物质中检测到28种物质, 醛类物质相对含量最高, 其次是醇类和酸类物质。EAG分析表明苯甲醛等13种竹笋挥发物的相对触角电位值显著高于对照, 以苯甲醛的EAG值最高。结果提示丛生竹笋梢高含量和特有的挥发物成分可能是吸引长足大竹象取食竹笋的重要物质, 长足大竹象对竹笋气味的识别很大可能是根据以苯甲醛为主要成分组成的、具有特定成分比例的化学指纹图谱。本研究为研制长足大竹象取食引诱剂提供了理论依据。     

华北大黑鳃金龟对桃树挥发物的电生理反应

【目的】华北大黑鳃金龟Holotrichia oblita是一种分布范围广,寄主种类多的农业害虫。本研究鉴定了桃树顶空挥发物中对华北大黑鳃金龟雌虫具有触角电生理活性的化合物,并探索桃树挥发物活性成分的剂量对华北大黑鳃金龟雌、雄虫触角电生理反应的影响。【方法】采用顶空吸附法收集桃树枝叶挥发物,用气相色谱 触角电位联用技术(GC-EAD)和气相色谱 质谱联用技术(GC-MS)鉴定有电生理活性的桃树挥发物成分,最后用触角电位(electroantennograph, EAG)技术测试这些挥发物在0.01,0.1, 1, 10和100 μg剂量下引起的华北大黑鳃金龟雌、雄成虫EAG反应。【结果】桃树挥发物中能引起华北大黑鳃金龟雌成虫触角电生理反应的成分有7种,包括Z-β-罗勒烯、顺-3-己烯乙酸酯、顺-3-己烯醇、壬醛、水杨酸甲酯以及2种未知化合物。所测试的对华北大黑鳃金龟雌成虫触角具有电生理活性的5种桃树挥发物成分在各剂量下都能引起雌、雄成虫明显的EAG反应。雌、雄虫对同种物质的EAG反应随着剂量的增加而有增大的趋势,并且雌虫的EAG反应显著高于雄虫。【结论】桃树挥发物中,罗勒烯、顺-3-己烯乙酸酯、顺-3-己烯醇、壬醛、水杨酸甲酯均能引起华北大黑鳃金龟雌、雄成虫的剂量依赖性的触角电生理反应,而且存在显著的性别差异。可利用这些化合物开展行为反应和田间诱集试验,进一步验证其对华北大黑鳃金龟的引诱活性。     

沙葱萤叶甲成虫触角感器超微结构及对沙葱挥发物的触角电位反应

【目的】明确沙葱萤叶甲Galeruca daurica成虫触角感器类型及对寄主植物挥发物的电生理反应,为进一步研究沙葱萤叶甲的化学感受机理奠定基础。【方法】使用扫描电子显微镜观察沙葱萤叶甲成虫触角上的感器类型;采用顶空动态吸附收集法采集寄主植物沙葱Allium mongolium的挥发物,利用气质联用仪测定沙葱主要挥发物组分,并利用触角电位技术(electroantennogram, EAG)测定沙葱萤叶甲成虫对这一寄主植物主要挥发物成分的电生理反应。【结果】沙葱萤叶甲成虫触角上分布的感器主要有5种类型,分别是毛形感器(sensilla trichodea, ST)、刺形感器(sensilla chaetica, SC)、锥形感器(sensilla basiconica, SB)、钟形感器(sensilla campaniformia, SCa)和B9hm氏鬃毛(B9hm bristles, BB)。沙葱挥发物主要由32种化合物组成,其中,含硫化合物占总量的49.3%。雌成虫对二烯丙基硫醚、二烯丙基二硫、顺-2-己烯-1-醇、2-己烯醛、苯甲酸甲酯和己醛6种化合物表现出较强的触角电位反...     

柑橘大实蝇信息素鉴定及EAG测试

[目的]分析和鉴定柑橘大实蝇Bactocera (Tetradacus) minax (Enderlein)信息素.[方法]采用顶空固相微萃取方法(HS-SPME)收集不同时间段柑橘大实蝇成虫挥发物,进一步结合气相色谱质谱联用技术(GC-MS)鉴定分析和触角电位技术(EAG)进行行为测试.[结果]N-(3-甲基丁基)乙酰胺为柑橘大实蝇成虫合成的一种挥发性化合物,且雌雄成虫都可以释放该化合物.触角电位技术测定不同日龄柑橘大实蝇成虫对1、10、100和1000 μg4种不同剂量N-(3-甲基丁基)乙酰胺的触角电位反应结果表明,柑橘大实蝇成虫对N-(3-甲基丁基)乙酰胺具有显著的触角电位反应,触角电位反应最适含量范围为大于100 μg,剂量以及剂量×性别会导致柑橘大实蝇显著的触角电位反应差异.[结论]N-(3-甲基丁基)乙酰胺可能为柑橘大实蝇信息素,这一结果可为柑橘大实蝇化学通讯研究和生物防治应用提供一定的理论基础.     

Electrophysiological and behavioral responses of a parasitic wasp to plant volatiles induced by two leaf miner species

In the present study, Y-tube olfactometric assays demonstrated that headspace volatile extracts collected from leaf miner-damaged, or artificially damaged, bean plants were more attractive to naive females of the parasitoid insect Opius dissitus than those collected from healthy plants. Headspace extracts from both Liriomyza huidobrensis and Liriomyza sativae second-instar larvae-damaged beans were analyzed by coupled gas chromatography-electroantennographic detection (GC-EAD). Of nine EAD-active volatiles identified, (3E)-4,8-dimethyl-1,3,7-nonatriene, (3Z)-hexenyl acetate, (syn)-2-methylpropanal oxime, and (syn)-2-methylbutanal oxime were the most abundant compounds that evoked significant electroantennogram (EAG) responses. Compounds (3Z)-hexen-1-ol, (anti)-2-methylbutanal oxime, linalool, beta-caryophyllene, and (3E,7E)-4,8,12-trimethyl-1,3,7,11-tridecatetraene also elicited clear EAG responses but were present in smaller amounts. Choice experiments in a Y-tube olfactometer indicated that synthetic versions of (3Z)-hexen-1-ol, 2-methylpropanal oxime, 2-methylbutanal oxime, 3-methylbutanal oxime, linalool, (E,E)-alpha-farnesene, and (3E,7E)-4,8,12-trimethyl-1,3,7,11-tridecatetraene were attractive individually, while (3Z)-hexenyl acetate and (3E)-4,8-dimethyl-1,3,7-nonatriene were unattractive at concentrations similar to those obtained from the headspace collection. Moreover, a blend of nine EAD-active volatiles was significantly more attractive relative to hexane (solvent). A mixture of oximes, tereponids, and green leaf volatiles may facilitate host location by female O. dissitus.     

Electrophysiological and behavioural responses of Scyphophorus acupunctatus (Col., Curculionidae) to Agave tequilana volatiles

Abstract:  The weevil Scyphophorus acupunctatus Gyllenhal (Col., Curculionidae) is the most important insect pest of cultivated agaves in Mexico. The objective of this study was to search for potential attractants for this weevil species from the freshly cut fleshy leaves of healthy Agave tequilana Weber var. Blue plants. Combined gas chromatography-electroantennography (GC-EAD) analysis of A. tequilana volatile extracts showed that seven peaks elicited antennal responses from males and females. Five of these peaks were identified by gas chromatography-mass spectrometry (GC-MS) as o -xylene, α -pinene, 3-carene, γ -terpinene and linalool. Electroantennogram (EAG) records of both sexes to 1-, 10- and 100- μ g-stimulus load of these five compounds showed that there was a dose effect on the EAG responses for linalool, 3-carene and α -pinene, but not for γ -terpinene and o -xylene. There were no sexual differences between male and female EAG responses to any of the compounds tested. Y-tube olfactometer bioassays of the compounds showed that males and females were attracted to α -pinene, 3-carene, γ -terpinene and/or linalool at doses of 1 and 10  μ g, while weevils were repelled by linalool at a dose of 100  μ g.     

Odor discrimination using insect electroantennogram responses from an insect antennal array

Insects have a highly developed olfactory sensory system, mainly based in their antennae, for the detection and discrimination of volatile compounds in the environment. Electroantennogram (EAG) response profiles of five different insect species, Drosophila melanogaster, Heliothis virescens, Helicoverpa zea, Ostrinia nubilalis and Microplitis croceipes, showed different, species-specific EAG response spectra to 20 volatile compounds tested. The EAG response profiles were then reconstructed for each compound across the five insect species. Most of the compounds could be distinguished by comparing the response spectra. We then used a four-antenna array, called a Quadro-probe EAG, to see if we could discriminate among odorants based on the relative EAG amplitudes evoked when the probe was placed in plumes in a wind tunnel and in a field. Stable EAG responses could be simultaneously and independently recorded with four different insect antennae mounted on the Quadro-probe, and different volatile compounds could be distinguished in real time by comparing relative EAG responses with a combination of differently tuned insect antennae. Regardless of insect species or EAG amplitudes, antennae on the Quadro-probe maintained their responsiveness with higher than 1 peak/s of time resolution.     

芒果不同部位挥发物及横线尾夜蛾对其触角电位反应

【背景】芒果横线尾夜蛾是严重危害芒果花序和嫩梢的重要害虫,其发生面积不断扩大,给我国南方芒果造成了巨大损失。【方法】通过气质联用仪（GC-MS）对芒果花、梢和成熟果实的挥发性物质及其相对含量进行分析;采用触角电生理方法,测定不同性别和交配状态的芒果横线尾夜蛾对12种芒果挥发物的触角电位反应。【结果】芒果3个部位的挥发性物质多为萜烯类化合物,主要物质（相对含量在O．06％以上）的数量分别为芒果花中22种、梢11种和成熟芒果13种。芒果花中含量较高的为d．水芹烯（43．64％）和异松油烯（29．33％）;芒果梢中α-古芸烯（23．63％）含量最高;成熟芒果中8-石竹烯含量最高,为29．47％。仅异松油烯和β-荜澄茄油烯在芒果3个部位中均被检测到,但其含量差异较大;异松油烯在花、梢、果实中的含量分别为29．33％、19．99％、0．59％,而β-荜澄茄油烯分别为3．51％、2．18％、6．05％。芒果横线尾夜蛾未交配雌、雄蛾和已交配雌、雄蛾对同一种化合物的触角电位相对值之间均无显著差异。4种成蛾对反-2-已烯醛的触角电位反应值与其他化合物相比均有显著差异。交配后雌、雄蛾的触角电位反应值均有所降低,但与交配前相比无显著差异。【结论与意义】筛选出的反-2-己烯醛可作为横线尾夜蛾信息素的增效物质,对性引诱剂的研制具有重要意义。本研究可为从化学信号角度阐明横线尾夜蛾寄主定向和雌、雄交配机理,以及其致害机理提供依据。     

Electroantennogram responses of Plutella xylostella (L.), to sex pheromone components and host plant volatile semiochemicals

The diamondback moth (DBM), Plutella xylostella (L.) (Lepidoptera: Plutellidae), is a notorious insect pest of cruciferous crops worldwide. Attract-and-kill strategies to manage the DBM based on insect pheromone and plant volatile semiochemicals have been explored and partially applied in the field. However, little is known about whether the attractant effects of insect pheromone and host plant volatile semiochemicals on insects are affected by insect age or volatile concentrations. Therefore, we examined the electroantennogram (EAG) responses of both DBM males and females varying in age and body size to a range of concentrations of seven host plant volatiles and two main female sex pheromone components. Our results showed that DBM age had no influence on EAG responses to trans-2-Hexenal, trans-2-Hexenol, Heptanal, cis-3-Hexen-1-ol and Z11-16:Ald and significantly influenced the EAG responses to Z11-16:Ac and three isothiocyanates. Age and sex had interaction effects on EAG responses of DBMs to some tested semiochemicals. DBM females had significantly stronger responses than males to all tested plant volatiles, and values were enhanced with increasing concentrations. Moreover, the degree of enhancement of EAG responses in females was larger than that in males for trans-2-Hexenal, trans-2-Hexenol, cis-3-Hexen-1-ol, 2-Phenylethyl isothiocyanate and Methyl isothiocyanate treatments. Body size seemed to have no influence on EAG responses. Our results might provide a theoretical basis for optimizing attract-and-kill strategies for insect pests.     

Chopper-stabilized gas chromatography-electroantennography: Part I. background, signal processing and example

A new method that can improve gas-chromatography-electroantennographic detection (GC-EAD) by orders of magnitude through a technique known as chopper stabilization combined with matched filtering in colored noise is presented. The EAD is a physiological recording from the antenna of an insect which can be used to find compounds in the GC effluent that the antenna is able to detect, having important applications for pest control and understanding of chemical communication in nature. The new method is demonstrated with whole-animal male Helicoverpa zea antennal preparations for detection of major pheromone component (cis-11-hexadecenal) and compared to results obtained using traditional EAD recording techniques. Results indicate that chopper stabilization under these circumstances can increase odorant detection performance by a factor of approximately 10(4) over traditional methods. The time course of the response of the antenna is also better resolved under chopped conditions. Although the degree of improvement is expected to vary with insect species, odor, and insect preparation, under most circumstances a more sensitive and robust GC-EAD instrument will result from the application of this technique.     

Electrophysiological responses of Atta sexdens rubropilosa workers to essential oils of eucalyptus and its chemical composition

The leaf-cutting ant Atta sexdens rubropilosa Forel, 1908 is the most harmful of the Eucalyptus pests, causing severe losses in wood production through defoliation. Various strategies have been tried and effort spent on the development of methods to control this pest, however no practical and environmentally acceptable one currently exists. In this work the chemical composition of the essential oil of seven Eucalyptus species was identified and the selectivity and sensitivity of antennal receptors of A. sexdens rubropilosa workers to the volatile compounds were determined using the electroantennographic technique (EAG and GC-EAD). Analysis by GC-EAD showed in E. cloesiana and E. maculata, respectively, seventeen and sixteen terpenes that elicited responses in ant workers' antennae, indicating the potential role of the essential oils as allelochemicals that determine the choice of the foraging material.     

苹褐带卷蛾成虫对主要寄主植物挥发物的触角电位反应和行为反应

【目的】苹褐带卷蛾Adoxophyes orana是危害苹果、桃、梨、枣等果树的一种重要昆虫。近年来在陕西苹果和红枣产区发生面积逐年扩大,危害日趋严重。本研究旨在明确寄主植物挥发物在苹褐带卷蛾成虫嗅觉通讯中的作用,为开发植物源引诱剂提供基础数据。【方法】利用触角电位(electroantennogram, EAG)仪测定苹褐带卷蛾2日龄未交配雌、雄成虫对51种寄主植物挥发性化合物的EAG反应;利用本实验室开发的小型蛾类昆虫嗅觉行为测试装置测定苹褐带卷蛾成虫对15种具有显著EAG反应的挥发性化合物的行为反应。【结果】EAG测试结果表明,苹褐带卷蛾成虫对不同挥发性化合物的EAG反应相对值有显著差异,雌、雄成虫对顺-3-己烯-1-醇、反-2-己烯-1-醇、1-己醇、1-庚醇、己醛、反-2-己烯醛、庚醛、辛醛、壬醛、乙酸丁酯、乙酸异戊酯和乙酸-顺-3-己烯酯的EAG反应较强。此外,雄成虫对3-甲基-1-丁醇、1-戊烯-3-醇、苯甲醛、丁酸丁酯、乙酸乙酯、三甲基巴豆酸乙酯、己酸乙酯、丙酸丁酯、异戊酸-顺-3-己烯酯、苯甲腈和柠檬腈的反应较强。在待测的51种挥发性化合物中,苹褐带卷蛾成虫对29种化...     

云斑天牛对10种植物挥发物的EAG和行为反应

【目的】为了探明对云斑天牛Batocera lineolata Chevrolat成虫具有行为导向的寄主植物挥发物对成虫行为反应的影响。【方法】采用触角电位(EAG)和"Y"型嗅觉仪技术,测定云斑天牛成虫(未交配雌虫、未交配雄虫、已交配雌虫)对10种寄主植物挥发物的电生理和行为反应。【结果】所选化合物在特定浓度下,均能引起云斑天牛成虫较为明显的EAG反应,浓度为100μg/μL时,云斑天牛成虫对所测试的植物挥发物的EAG反应最强,其中未交配雌、雄及已交配雌虫对(Z)-3-己烯-1-醇的EAG反应相对值均为最大,分别达2.35、2.31和2.22。行为反应生测中,(Z)-3-己烯-1-醇对未交配雌、雄虫可产生明显的引诱作用,芳樟醇则对两者具有明显的趋避作用,而已交配雌虫对所有化合物均无明显趋性。【结论】试验结果可为开发对其有效的林间引诱剂提供一定的理论依据。