暗黑鳃金龟对性信息素的触角电生理及行为反应

为了分离鉴定暗黑鳃金龟的性信息素成分并对其功能进行验证,本研究对比摸索了3种触角的处理方法,并进行同一浓度（30 ng·μL^-1）二元混合物、不同配比的触角EAG测试。结果包括: 建立了一种鳃叶状触角的触角电位（EAG）测定方法,即将暗黑鳃金龟Holotrichia parallela触角的各鳃叶用针分离开后进行测定,这种方法测得的触角电位反应值较高。雄虫触角对L-异亮氨酸甲酯和(R)-(-)-芳樟醇为7∶1的二元混合物的反应值最高,和暗黑鳃金龟雌虫性信息素25个腺体提取液的EAG反应相当;对同一配比（7∶1）不同剂量刺激液的EAG测试表明,雄虫对浓度为30 ng·μL^-1的二元混合物刺激液的反应值最高。嗅觉反应结果进一步证实,试虫对L-异亮氨酸甲酯和(R)-(-)-芳樟醇7∶1的选择最高,选择系数达93.3%。研究结果为利用性信息素防治暗黑鳃金龟技术的开发提供了基础。     

Investigation of PAS and CNBH domain interactions in hERG channels and effects of long-QT syndrome-causing mutations with surface plasmon resonance

Human ether-á-go-go-related gene (hERG) channels are key regulators of cardiac repolarization, neuronal excitability, and tumorigenesis. hERG channels contain N-terminal Per-Arnt-Sim (PAS) and C-terminal cyclic nucleotide-binding homology (CNBH) domains with many long-QT syndrome (LQTS)-causing mutations located at the interface between these domains. Despite the importance of PAS/CNBH domain interactions, little is known about their affinity. Here, we used the surface plasmon resonance (SPR) technique to investigate interactions between isolated PAS and CNBH domains and the effects of LQTS-causing mutations R20G, N33T, and E58D, located at the PAS/CNBH domain interface, on these interactions. We determined that the affinity of the PAS/CNBH domain interactions was ∼1.4 μM. R20G and E58D mutations had little effect on the domain interaction affinity, while N33T abolished the domain interactions. Interestingly, mutations in the intrinsic ligand, a conserved stretch of amino acids occupying the beta-roll cavity in the CNBH domain, had little effect on the affinity of PAS/CNBH domain interactions. Additionally, we determined that the isolated PAS domains formed oligomers with an interaction affinity of ∼1.6 μM. Coexpression of the isolated PAS domains with the full-length hERG channels or addition of the purified PAS protein inhibited hERG currents. These PAS/PAS interactions can have important implications for hERG function in normal and pathological conditions associated with increased surface density of channels or interaction with other PAS-domain-containing proteins. Taken together, our study provides the first account of the binding affinities for wild-type and mutant hERG PAS and CNBH domains and highlights the potential functional significance of PAS/PAS domain interactions.     

豆野螟成虫触角对植物挥发性和性信息素化合物的触角电位反应

以顺3-己烯乙酸酯为标准化合物,研究了豆野螟雌、雄成虫触角对不同浓度的植物挥发性和性信息素化合物组分的触角电位（EAG）反应.结果表明：豆野螟雌、雄成虫触角对植物挥发性气味具有强烈的EAG反应,但在大部分浓度下,雌、雄虫的反应没有显著差异.在测试的9种植物挥发性化合物中,雌、雄虫对高浓度反-2-己烯醛刺激的相对EAG反应的最大值分别达到250%和260%,且雌、雄虫触角对这种化合物的EAG反应的形状也有很大差异;但对豇豆花气味中的柏木烯和柏木脑没有明显的EAG反应.雌、雄蛾对性信息素都有明显反应,且雌、雄虫的反应存在显著差异,雄虫触角对性信息素化合物主要组分（反10,反12）-十六碳二烯醛的EAG反应的最大值达到250%,远超过雌虫反应的最大值(53%).雌、雄成虫这种神经感觉反应的差异反映了它们在感器结构、功能和行为上的差异.     

与中红侧沟茧蜂生境与寄主定位相关的玉米及棉铃虫幼虫体表挥发性成分的提取与鉴定

【目的】鉴定玉米Zea mays L.和棉铃虫Helicoverpa armigera (Hübner)幼虫体表挥发性成分中对中红侧沟茧蜂 Microplitis mediator (Haliday)具有生境与寄主定位作用的信息化合物,从化学生态学的角度研究玉米-棉铃虫-中红侧沟茧蜂三重营养关系,解释中红侧沟茧蜂寻找寄主的过程中的信息识别机制,为害虫的综合防治的“推-拉”方法提供一定的理论基础。【方法】利用触角电位仪(EAG)、触角电位联用仪(GC-EAD)、气质联用仪(GC-MS)及“Y”型嗅觉仪确定玉米和棉铃虫幼虫体表提取物的信息化合物。在室内利用玉米以及棉铃虫幼虫体表挥发物标准品化合物以及模拟混合物,使用“Y”型嗅觉仪进行中红侧沟茧蜂成虫行为反应试验。【结果】玉米挥发物中有11种化学成分,棉铃虫幼虫体表挥发物中有6种化学成分对中红侧沟茧蜂的触角具有电生理活性,其中4种成分在两种挥发物中都存在。室内行为反应试验发现：与正己烷对照相比,玉米的模拟组分对雌、雄蜂均表现出显著（P<0.05）的诱引作用;棉铃虫1龄幼虫体表模拟组分对雌蜂具有极显著的诱引作用（P<0.01）,对雄蜂具有显著的诱引作用（P<0.05）;棉铃虫2龄幼虫体表模拟组分对雌蜂具有显著的诱引作用（P<0.05）。【结论】本研究证明了玉米以及棉铃虫幼虫体表挥发物中分别存在11种（庚醛、2-己醇、1-己醇、1-辛烯-3-醇、壬醛、葵醛、苯甲醛、反式-2-壬烯-1-醇、己酸、苯基乙醇、月桂醇）和6种（2-己醇、己酸乙酯、1-己醇、壬醛、辛酸乙酯、癸醛）中红侧沟茧蜂生境及寄主定位的化学信息物质。     

Identification and Knockdown of the Olfactory Receptor (OrCo) in Gypsy Moth,Lymantria dispar

The gypsy moth, Lymantria dispar, is an important economic pest that causes large-scale damage to forests worldwide. Because of its important role in initiating and controlling insect behavior, olfaction—and olfaction-based pest management—has drawn increasing attention from entomologists. In this study, we identified the gene that encodes the olfactory receptor co-receptor (OrCo). Through amino acid sequence alignment, we found that LdisOrCo shares high identity with other OrCo proteins from different insect orders. Next, we performed RNA-interference (RNAi) to assess the role of OrCo in olfaction. Electroantennographic assays showed that after RNAi, the average value of males'' response to sex pheromones was 0.636 mV, significantly lower than that of the positive control (average = 1.472 mV). Females showed no response to sex pheromones before or after RNAi. Finally, quantitative PCR showed a strong decrease in the expression of OrCo after RNAi, by ~74% in males and by 23% in females relative to the positive controls. These results indicate that OrCo is not only critical to odor recognition, but it may also represent a new target for development of semiochemicals that can influence insect behavior.     

Isolation of sex pheromone mimic of the American cockroach by monitoring with male/female ratio in electroantennogram

The electroantennogram (EAG) technique was applied as a monitoring tool to isolate germacrene-D, a sex pheromone mimic of the American cockroach. Large values of the male/female ratio, and its index derived from EAG responses of adult males and females to a standard, indicated significant amounts of germacrene-D in plant fractions. Monitoring with behavioural assays and gas chromatographic analysis during the isolation confirmed that the EAG technique using the male/female index was a good indicator in the isolation of sexually active components.     

Electrophysiological and behavioral responses of Asian and European honeybees to pear flower volatiles

Honeybee pollination behavior is influenced by flower volatiles, which honeybees sense via olfactory receptors. Honeybees are only weakly attracted to pear flowers. To investigate the potential reasons, we extracted and determined the floral volatile compounds from three pear cultivars (Su, Ya, and Xuehua) using headspace solid-phase micro-extraction (HS-SPME) and gas chromatography-mass spectrometry (GC–MS). The effects of pear flower volatiles on the Asian honeybee (Apis cerana cerana Fabricius) and the European honeybee (Apis mellifera ligustica Spinola) were determined by electroantennogram (EAG) assays and behavioral tests in a three-arm olfactometer. Among the 76 flower volatiles detected with GC–MS, 21 were found in all three pear cultivars, accounting for approximately 70% of the total volatile content. 3-Methyl-1-butanol and (+)-limonene volatiles had the highest relative content. Five compounds elicited strong EAG responses in both bee species: 2-methylbutyraldehyde, 1-nonanal, 6-methyl-5-hepten-2-one, 3-methyl-1-butanol, and (+)-limonene. Neither bee species showed positive taxis to these volatiles. In behavioral tests, A. mellifera ligustica showed a low preference for 6-methyl-5-hepten-2-one (20%, 400 µg/µL) and 2-phenethyl alcohol (16.7%, 400 µg/µL). Apis cerana cerana showed a low preference for 6-methyl-5-hepten-2-one (6.7%, 400 µg/µL) and 1-nonanal (10%, 400 µg/µL), whereas its preferences for 3-methyl-1-butanol (43.3%, 400 µg/µL) and α-farnesene (40%, 400 µg/µL) were similar to that for the control. Therefore, a lack of attractive volatile compounds could explain why honeybees are only weakly attracted to pear flowers. Therefore, to achieve acceptable pollination in pear orchards, we suggest using flower-scent sugar syrup feeding and a saturation pollination strategy.     

Potassium channels as tumour markers

An increasing number of ion channels are being found to be causally involved in diseases, giving rise to the new field of "channelopathies". Cancer is no exception, and several ion channels have been linked to tumour progression. Among them is the potassium channel EAG (Ether-a-go-go). Over 75% of tumours have been tested positive using a monoclonal antibody specific for EAG, while inhibition of this channel decreased the proliferation of EAG expressing cells. The inhibition of EAG is accomplished using RNA interference, functional anti-EAG1 antibodies, or (unspecific) EAG channel blockers. Fluorescently labelled recombinant Fab fragments recognizing EAG allow the distribution of EAG to be visualized in an in vivo mouse tumour model.