挥发性植物源害虫引诱剂筛选与混配方法的新视角

成虫期是农业害虫世代发育链中容易实施人为干预的薄弱环节,利用植物源引诱剂诱杀成虫具有微量高效、兼容配套、环境友好、不易产生抗性等优点。此前已经筛选出了一些简单配方,但其害虫诱杀量占自然种群的比例以及诱杀选择性还未达到令人满意的水平。传统化学生态学技术针对植物源害虫引诱剂的筛选存在两个难题,一是如何将组织程度较高的植物挥发物谱合理地分解为若干子系统,然后对其功能进行有机的耦合;二是针对数量众多的电生理或行为活性成分,如何设计引诱剂配方并进行整体优化。在这篇综述中,作者指出了传统化学生态学某些技术环节的不足之处;综述了近年来国内外植物源害虫引诱剂筛选与应用的新思路以及人类调香术对于引诱剂研制的启示;给出了电生理活性成分的正交组合测定法,以及引诱剂大田配方设计的关键方法——配方均匀设计的原理和案例。     

桔小实蝇对几种寄主挥发物的触角电位反应

桔小实蝇寄主范围广,可危害芒果、柑橘、咖啡和红辣椒等250多种水果和蔬菜,现已成为中国、东南亚、印度次大陆和夏威夷群岛一带的危险性果蔬害虫。本文利用触角电位(electroantennogram,EAG)技术,测定了桔小实蝇雌雄成虫对8种寄主挥发物标准化合物(1,8桉叶素、α-蒎烯、β-石竹烯、柠檬醛、香草醛、乙醇、芳樟醇和月桂烯)的EAG反应。结果表明:上述标准化合物均能引起EAG反应,EAG反应值随着浓度增大(0.1μg/μL、1μg/μL、10μg/μL和100μg/μL)而显著不同。当柠檬醛和月桂烯浓度为100μg/μL时,桔小实蝇雌虫交配前、后的EAG反应值存在显著性差异。浓度为100μg/μL的1,8桉叶素对桔小实蝇交配后雌虫和雄虫的刺激反应存在显著性差异。桔小实蝇交配前雌虫和雄虫对浓度为100μg/μL乙醇的EAG反应值存在显著性差异。     

松阿扁叶蜂对不同树种挥发物的触角电位反应

用MS302生物信号记录分析系统,以顶端记录法测定松阿扁叶蜂成虫对不同树种挥发性物质的EAG反应。结果显示,该蜂成虫对其嗜好的油松、赤松、华山松挥发物的EAG反应敏感,雌成虫放电频率分别为45.2次/s、42.1次/s、42.3次/s,电位值依次为1 022 mV、1.139 mV、0.706 mV, 但对其非寄主树种的侧柏挥发物反应微弱,对空气无反应。雌成虫对油松挥发物的不同气流量17 mL/s、17 mL/s、85 mL/s的放电频率和电位值分别为24.8次/s、45.2次/s、46.6次/s和0.533 mV、1.022 mV、1.316 mV,表明EAG反应程度因挥发物气流量浓度不同而变化。触角电位与选择性行为的测试结果基本一致,表明松阿扁叶蜂在寄主选择过程中,触角的嗅觉功能起着重要作用。     

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...     

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。     

美洲大蠊对多种化合物的触角嗅觉电位反应

用多种化合物的气味制激美洲大蠊(Periplaneza americana)触角,诱导、记录触角电位(EAG)反应.试验了各类化合物154种,其中53%能诱发出明显的EAG反应.绝大多数EAG反应是负相慢波,波幅多数为0.5—2mV,时程1.5—3秒.苯等少数化合物能诱发正相EAG反应.剂量-反应曲线大致呈“S”形,还可见适应现象.萜类化合物和挥发性植物油类诱发EAG反应的作用最强,其次是酮、醇、酯、胺类化合物.化合物的含氧功能基团和空间构型与EAG反应的诱发有一定的关系.可能是同一种化合物对不同种的感受器都可能诱发EAG反应.一般性化合物诱发雌、雄、成、幼虫EAG反应的差异不大,但雌虫类提取物仅对雄性成虫触角诱发较强的EAG反应.一些已知的驱、诱、杀虫剂也能诱发EAG反应,但仅表现为对一般化合物的反应特征.     

烟蚜茧蜂对蚜虫信息素及烟草挥发物的触角电位反应

利用触角电位(EAG)技术测定了烟蚜茧蜂(Aphididius gifuensis)对蚜虫性信息素(荆芥醇和荆芥内酯)和报警信息素(反-β-法尼烯)以及烟草挥发性物质的电生理反应.结果表明:雌蜂和雄蜂对蚜虫性信息素、报警信息素、烟草挥发物均有反应,但雌雄蜂对这些化学信息物质的嗅觉敏感性存在差异,雌蜂对荆芥醇、荆芥内酯、顺-3-己烯-1-醇、反-2-己烯醛、水杨酸甲酯和里那醇的EAG反应都大于雄峰,证明雌雄蜂在利用寄主栖境中信息化学物质方面存在不同的策略,它们分别识别了不同的有关寄主的化学指纹图;烟蚜茧蜂对烟草气味组分具有不同的敏感性,对绿叶气味组分的EAG反应要强于对萜类化合物的反应,绿叶气味组分很有可能在其寻找桃蚜的过程中发挥远距离定向作用.     

苹毛丽金龟对植物挥发物成分的触角电位及行为反应

为筛选适宜的苹毛丽金龟Proagopertha lucidula (Faldermann)引诱剂化学物质, 对已报道的金龟子科两性引诱剂单一组分进行了触角电位（EAG）和Y型嗅觉仪中的嗅觉反应测定。结果表明: 在10 μg刺激剂量下测试的11 种植物挥发物中, 只有顺-3-己烯-1-醇、香叶醇、茴香脑和丁香酚有明确的EAG反应值, 其他7种化合物具有弱反应值。剂量反应曲线表明, 具有明确的EAG活性的4种化合物激发的反应强度与化合物的剂量呈正相关。在Y型嗅觉仪中, 顺-3-己烯-1-醇、香叶醇、茴香脑和丁香酚4种化合物在1, 10, 100 μg剂量下的行为反应百分率都显著高于对照溶剂正己烷, 且雌雄之间无显著差异。该研究结果可为开发苹毛丽金龟田间引诱剂奠定基础。     

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。     

飞蝗对两种禾本科植物挥发物的触角电位和行为反应

【目的】检测飞蝗对2种禾本科植物挥发物——癸醛和芳樟醇的触角电位及行为反应,为了解飞蝗的嗅觉机制提供依据。【方法】首先,使用触角电位仪检测飞蝗4龄若虫对癸醛和芳樟醇4个剂量的触角电位反应。其次,使用嗅觉行为仪检测正常条件下4龄飞蝗对癸醛和芳樟醇不同剂量的行为反应。最后,采用嗅觉行为仪检测饥饿和饱食状态下的虫体对癸醛和芳樟醇的行为反应。【结果】4龄飞蝗对癸醛和芳樟醇的EAG值随着剂量增加呈先升高后降低的趋势,均在500μg达到最高值,且在该剂量下,雌雄虫之间的EAG值存在显著差异。正常条件下,1 000μg芳樟醇对飞蝗雌雄虫均有显著的引诱作用,1 500μg癸醛和芳樟醇显著引诱雄虫,500μg癸醛显著引诱雌虫。经饥饿处理后,雌虫被500μg和1 000μg癸醛显著引诱,雄虫被250μg癸醛显著引诱,雌雄虫均被500μg的芳樟醇显著引诱。经过饱食处理后,250μg和500μg的芳樟醇显著引诱雌虫。芳樟醇对饥饿状态下飞蝗雄虫的引诱率均高于饱食状态,在500μg时,引诱率存在显著差异。【结论】飞蝗对这2种植物挥发物的触角电位反应存在剂量效应,雌雄虫触角电位在最佳剂量下存在显著差异。不同剂量的癸醛和芳樟醇可对飞蝗产生不同的引诱效果,饥饿状态下的虫体更易被芳樟醇所吸引。     

On the distribution of the common cabbage butterfly,Pieris rapae crucivora,in the field

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Contributions from the Department of Biology, Miyazaki University, No. 45.     

Pattern of spatial distribution of the common cabbage butterfly,Pieris rapae in a cabbage farm

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Contributions from the Entomological Laboratory, Kyoto University, No. 203.     

Purification and some properties of beta-N-acetyl-D-glucosaminidase from the cabbage butterfly (Pieris rapae)

A beta-N-acetyl-D-glucosaminidase (NAGase) from the cabbage butterfly (Pieris rapae) was purified. The purified enzyme was a single band on polyacrylamide gel electrophoresis and the specific activity was determined to be 8715 U/mg. The molecular weight of whole enzyme was determined to be 106 kDa by gel filtration, and the result of SDS-PAGE showed that the enzyme was a heterodimer, which contained two subunits with different mass of 59.5 and 57.2 kDa. The optimum pH and optimum temperature of the enzyme for the hydrolysis of p-nitrophenyl-N-acetyl-beta-D-glucosaminide (pNP-NAG) were investigated to be at pH 6.2 and at 42 degrees C, respectively, and the Michaelis-Menten constant (K(m)) was determined to be 0.285 mM at pH 6.2 and 37 degrees C. The stability of the enzyme was investigated and the results showed that the enzyme was stable at the pH range from 4.0 to 9.0 and at the temperature below 45 degrees C. The activation energy was 83.86 kJ/mol. The reaction of this enzyme with pNP-NAG was judged to be Ordered Bi-Bi mechanism according to the inhibitory behaviors of the products. The ionization constant, pK(e), of ionizing group at the active site of the enzyme was found to be 5.20 at 39.0 degrees C, and the standard dissociation enthalpy (DeltaH(o)) was determined to be 2.18 kcal/mol. These results showed that the ionizing group of the enzyme active center was the carboxyl group. The results of chemical modification also suggested that carboxyl group was essential to the enzyme activity. Moreover, Zn(2+), Hg(2+), Cu(2+) had strongly inhibitory effects on the enzyme activity.     

Process generating the ditribution pattern of eggs of the common cabbage butterfly,Pieris rapae crucivora

The process generating the negative binomial in the distribution pattern of eggs of the common cabbage butterfly, Pieris rapae crucivora, was investigated by releasing the female adults in a net house where cabbages were planted. The distribution of butterflies visited and laid an egg or more per plant followed thePoisson series under the uniform light condition, while that of eggs laid per visit conformed to the logarithmic distribution. From these results, it may be concluded that the negative binomial arises from compounding of thePoisson and the logarithmic distribution. The observed frequency of eggs found per plant fitted to the negative binomial with parameter thus computed theoretically. The change in the degree of aggregation with the increase of the parental density was considered in connection with the above results.     

Associative learning in the ovipositional behaviour of the cabbage butterfly, Pieris rapae

ABSTRACT. When tested with familiar and novel oviposition sites, gravid female Pieris rapae (L.) tended to land on sites of the same appearance as those they had experienced 24 h earlier. This acquired preference could be reversed by subsequent e xperience. An ovipositional preference, shown by groups of insects, paralleled the landing preferences of single insects. These influences of experience were shown with intact cabbage plants of different kinds, with discs cut from the leaves of different kinds of cabbage, and with discs of blotting paper of different shades of green and blue, wetted with sinigrin solution or water. A single landing by a gravid female on paper wetted with sinigrin solution induced a landing preference for the same colour of paper in a choice test made 24 h later. Gravid females thus associated a favourable chemical stimulus of oviposition with the appearance of the acceptable site. Such learning by trial and error would facilitate host-selection from a distance. In the laboratory it induced oviposition on 'neutral' sites of the same appearance as 'host' sites.     

Influence of parental density on the distribution pattern of eggs in the common cabbage butterfly,Pieris rapae crucivora

To investigate the relation between the distribution pattern of eggs and the parental density in the common cabbage butterfly, Pieris rapae crucivora, the countings of egg number per plant were made on both cabbage plants cultivated in the farm and planted in the net house in which the female butterflies were released at various densities. The frequency distribution of eggs fits well to the negative binomial excepting the cases where they agree withPoisson series, and the degree of aggregation expressed as the reciprocal of the parameter, 1/k, tends to decrease as the egg or parental density increases. At the same parental density, however, the distribution of eggs can be described by the negative binomial with a common parameter, k_c, regardless of the difference in the density of laid eggs. In the case where a single female butterfly lays eggs, the spatial pattern of egg distribution is always lean, while its frequencies conform toPoisson or the negative binomial series. This lean changes toward patchy with increasing the parental density. From these results, it is concluded that the degree of aggregation in the distribution of eggs decreases with the increase of the parental density.     

Avoidance of maladaptive,precocious copulation in the cabbage white butterfly,Pieris rapae crucivora

In the cabbage white butterfly, Pieris rapae crucivora, copulation hinders normal expansion and hardening of the wings of newly emerged females. The resulting permanent wing deformation makes it impossible for females to fly and therefore, to find an oviposition site and nectar sources. An attempt was made to clarify whether the newly emerged female butterfly avoids copulation. Observation of wing expansion and hardening reveals that the wings are fully expanded and hard by 20–30 min after emergence. In the field, presentation of females with soft wings to males shows that males will attempt to copulate with these females. However, newly emerged females prevent successful completion of copulation by assuming the mate refusal posture, and thereby avoid a potentially maladaptive copulation. The discussion focusses on the question as to why females and not males avoid early copulation.     

Constituents of host- and non-host plants deterring oviposition by the cabbage butterfly, Pieris rapae

Oviposition by Pieris rapae L. on cabbage was deterred by homogenized cabbage tissue sprayed onto intact plants. Ether extracts of cabbage also were deterrent, but water extracts were not. Hexane extracts of other host plants deterred oviposition and water extracts had little or no effect. Polar as well as non-polar extracts of non-host plants inhibited oviposition. Polar deterrents in the non-host crucifers, Erysimum cheiranthoides and Capsella bursa-pastoris may explain the avoidance of these plants by P. rapae. Chemical deterrents, as well as stimulants, apparently play a major role in the acceptance or rejection of plants as hosts by ovipositing female butterflies.

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Zusammenfassung Die Eiablage von Pieris rapae auf Kohl wurde gehemmt von einem Homogenisat aus Kohlblättern, das auf intakte Pflanzen gesprüht wurde. Ether-Extrakte der Kohlblätter wirkten im Gegensatz zu Wasserextrakten abschreckend. Ebenso zeigten Hexanextrakte anderer Wirtspflanzen eine hemmende Wirkung auf die Eiablage, während Wasserextrakte anderer Wirtspflanzen keine Wirkung aufwiesen. Dagegen aber verhinderten sowohl polare als auch unpolare Extrakte von Nicht-Wirtspflanzen die Eiablagen. Polare, abschreckende Substanzen in den Nicht-Wirtspflanzen Erysimum cheiranthoides und Capsella bursa-pastoris (Cruciferae) könnten Ursache dafür sein, dass P. rapae diese Pflanzen meidet. Chemische Substanzen mit sowohl abschreckender als auch stimulierender Wirkung spielen offensichtlich eine bedeutende Rolle für legebereite Schmetterlingsweibchen bei der Auswahl von Pflanzen als geeigneten Eiablageplatz.