Cross-modal interaction between visual and olfactory learning in Apis cerana

The power of the small honeybee brain carrying out behavioral and cognitive tasks has been shown repeatedly to be highly impressive. The present study investigates, for the first time, the cross-modal interaction between visual and olfactory learning in Apis cerana. To explore the role and molecular mechanisms of cross-modal learning in A. cerana, the honeybees were trained and tested in a modified Y-maze with seven visual and five olfactory stimulus, where a robust visual threshold for black/white grating (period of 2.8°–3.8°) and relatively olfactory threshold (concentration of 50–25 %) was obtained. Meanwhile, the expression levels of five genes (AcCREB, Acdop1, Acdop2, Acdop3, Actyr1) related to learning and memory were analyzed under different training conditions by real-time RT-PCR. The experimental results indicate that A. cerana could exhibit cross-modal interactions between visual and olfactory learning by reducing the threshold level of the conditioning stimuli, and that these genes may play important roles in the learning process of honeybees.     

引入西方蜜蜂对中蜂的危害及生态影响

作者阐述自1896年中国引进西方蜜蜂Apis melliferaL.的优良品种如意大利蜂Apis mellifera ligusticaSpinola和喀尼阿兰蜂Apis mellifera Carnica Pollmann以来,使当地的东方蜜蜂Apis cerana F.受到严重危害,其分布区域缩小75%以上,种群数量减少80%以上。使山林植物授粉总量减少,导致植物多样性减少。文中提出: 建立自然保护区保存本地蜜蜂遗传特性,和采用基因转移等技术,培育具有西方蜜蜂优良生产性能的中蜂新品种,逐步恢复中蜂的种群数量。     

常用杀虫剂噻虫嗪及其4种混配制剂对中华蜜蜂的生存风险分析

噻虫嗪及其混配制剂是防治刺吸式口器害虫的常用药剂,但对中华蜜蜂的生存风险尚未明确。在实验室条件下通过模拟中华蜜蜂授粉期间农药暴露的3种方式(蜂体接触、取食接触、植株接触),评估了噻虫嗪及其4种混配制剂,在田间最高推荐剂量下对中华蜜蜂采集蜂的毒性。结果表明,无论哪种暴露方式,噻虫嗪及其混配制剂均显著影响中华蜜蜂采集蜂的生存,且噻虫嗪混配制剂毒性高于单剂。在直接喷洒蜂体处理中,25%噻虫嗪·异丙威可湿性粉剂表现出最高毒性,其致死中时间(LT_(50)值)为1.23 h;在摄入和接触植株残留处理中,40%氯虫·噻虫嗪水分散粒剂和25%噻虫嗪·异丙威可湿性粉剂均表现出最高毒性,LT_(50)值分别为2.09、6.75 h和2.15、6.77 h。在3种暴露方式下,25%噻虫嗪水分散粒剂的LT_(50)值分别为6.50、17.24和8.90 h,明显低于对照。因此建议蜜蜂授粉期间禁止施用噻虫嗪及其混配制剂,为更好地推进蜜蜂授粉与绿色防控技术的研究与示范提供安全保障。     

中华蜜蜂嗅觉受体AcerOrco的表达及定位分析

【目的】通过对中华蜜蜂Apis cerana cerana嗅觉受体AcerOrco的表达及蛋白定位分析,阐明AcerOrco的表达特性,以期为进一步探索其功能提供理论依据。【方法】分别通过qRT-PCR技术和Western blot技术对中华蜜蜂气味受体AcerOrco mRNA及蛋白在内勤蜂和采集蜂5个组织部位(触角、头、胸、腹和足)中的相对表达量进行分析,采用免疫组化技术对AcerOrco蛋白的表达进行定位分析。【结果】定量结果显示,AcerOrco转录本在内勤蜂和采集蜂各组织中均有表达,其中触角中的表达量最高;该基因在采集蜂各组织中的表达量普遍高于内勤蜂。AcerOrco受体蛋白在内勤蜂和采集蜂各组织中也均有表达,在触角和头部的表达量明显高于其他组织。定位结果显示,在内勤蜂触角中,AcerOrco主要在毛形感器中表达,板形感器中表达不明显;在采集蜂触角的板形感器和毛形感器中均有表达,但毛形感器中的表达更多一些。【结论】获得了AcerOrco mRNA及其蛋白在内勤蜂和采集蜂各组织中的表达特性,并将这一蛋白表达定位于工蜂触角的毛形感器和板形感器中。     

The origin and genetic diversity of the yellow-legged hornet,Vespa velutina introduced in Japan

Insectes Sociaux - Biological invasions have severe economic and ecological impacts. The yellow-legged hornet, Vespa velutina was originally distributed throughout most of the Oriental region, but...     

Distribution,spread, and impact of the invasive hornet Vespa velutina in South Korea

Hornets (Vespa spp) are top insect predators that can control pests, but their venomous stings and defensive behavior cause numerous human deaths throughout Asia. Hornets usually inhabit rural areas which reduces potential conflict with humans. In 2003, the invasive hornet, Vespa velutina, arrived in southern Korea (Yeongdo region) and became established. It is currently spreading northwards at a rate of 10–20 km per year. Despite originating in tropical/subtropical areas of Indo-China, its nesting biology and life cycle in South Korea are similar to those found throughout its native range, with mature colonies containing 1000–1200 adults. In 7 years, V. velutina has become the most abundant hornet species in Southern Korea by displacing native Vespa species such as V. simillima, which has a similar nesting biology. We also found a significant positive correlation between the abundance of V. velutina and the degree of urbanization, indicating that this invasive species was well adapted to urban environments. This was supported by our finding that 41% of emergency call-outs (119 Rescue Services) to deal with social wasps/hornet problems were due to V. velutina, which was twice as high as the number of calls about the next most abundant species. The rapid spread of V. velutina across southern Korea indicates that this species will continue to spread north-westward in the Korean peninsula and will become a major problem as more people and beekeepers come into contact with this aggressive invasive hornet.     

Spatial coincidence of cues in visual learning by the honeybee (Apis mellifera)

The discrimination of patterns was studied in a Y-choice chamber fitted with a transparent baffle in each arm, through which the bees had a choice of two targets via openings 5cm wide. The bees see the positive (rewarded) and the negative (unrewarded) targets from a fixed distance. The patterns were bars (subtending 22 degrees x5.4 degrees at the point of choice) presented in one-quarter of each target. The bars were moved to a different quarter of the target every 5min, to make the location of black useless as a cue. A coincident presentation is when the bar on the left target is on the same side of the target as the bar on the right target. The bees learn the orientation cue when the presentation is coincident but otherwise cannot learn it. This experiment shows that bees do not centre their attention on the individual bars, otherwise they would always discriminate the orientation. Centring the target as a whole precedes learning. Having learned with the bar on one side of the targets, bees do not recognize the same cue presented on the other side. A separate orientation cue can be learned on each side. A radial/tangential cue is preferred to a conflicting orientation cue.     

北京中华蜜蜂的保护与利用

阐述目前中华蜜蜂Apis cerana Fabricius的分布及其生态影响,并针对北京地区中蜂现状,提出从中蜂饲养技术培训、饲养繁育示范、多元开发利用、建立专业养殖乡及发挥中蜂优势、发展山区养蜂事业等5个方面入手,保护与利用中华蜜蜂,逐步恢复中蜂的数量。     

The parasitoid fly Exorista japonica uses visual and olfactory cues to locate herbivore‐infested plants

Some parasitoid flies exploit odors derived from plants as olfactory cues for locating the food plants of host insects, but the role of visual cues associated with plants remains largely unknown. The generalist tachinid Exorista japonica Townsend (Diptera: Tachinidae) is attracted to odors derived from maize plants [Zea mays L. (Poaceae)] infested by the larvae of Mythimna separata (Walker) (Lepidoptera: Noctuidae). In this study, we examined the effects of visual parameters on the olfactory attraction of female flies to host‐infested plants. A paper plant model of one of four colors (blue, green, yellow, or red) was placed in front of a host‐infested plant, which was hidden behind a mesh screen in a wind tunnel. The landing rate of females was significantly higher on the green plant model than on the other three models. When an achromatic plant model of one of four gray scales (white, light gray, dark gray, or black) was tested, the response rate of females was significantly higher towards the white model and decreased as the brightness of models decreased. Few female flies responded to the green plant model without odors of the host‐infested plants. When the four color plant models were placed together in a cage filled with odors of host‐infested plants, females remained significantly longer on the green model than on the other three models. These results showed that E. japonica females preferred the color green when odors of the host‐infested plants were present and suggest that E. japonica uses visual as well as olfactory cues to locate the host habitat.     

不同组配的蜜蜂信息素对中华蜜蜂生产及繁殖性能的影响

【目的】为了探究蜜蜂信息素对中华蜜蜂Apis cerana cerana的生产及繁殖性能的影响。【方法】以蜂蜡为载体,选择4种蜂王上颚腺信息素（9-ODA:9-HDA:HOB:HVA）与蜜蜂幼虫饥饿信息素（β-罗勒烯）按照一定比例组配了4种强蜂素挂片（T1-1组、T1-2组、T2-1组、T2-2组）,同时设置一个纯蜂蜡挂片作为空白对照组（CK组）。将2种组配强蜂素（T1-1、T1-2）分别置于特殊密闭容器瓶中,15、30和45 d后用捕集针抽取密闭容器瓶气体,利用气相-质谱联用系统测定2种组配强蜂素（T1-1、T1-2）中β-罗勒烯挥发含量。并系统研究了4种组配强蜂素对无王群急造王台以及对有王群的蜂蜜产量、群势和封盖子数影响。【结果】在15、30和45 d时,2种组配强蜂素中β-罗勒烯挥发量差异不显著（P> 0.05）,即2种组配强蜂素中β-罗勒烯都能稳定释放;与对照组相比,T1-2组封盖王台出现时间显著延迟（P<0.05）,推迟时间2.17d,但4个实验组间出现封盖王台出现时间差异不显著（P>0.05）;T1-1组蜂蜜产量和封盖子数显著高于对照组（P <0.05）,蜂蜜产量和封盖子数量分别30.18%和30.00%;T1-2组蜂群群势显著强于对照组（P <0.05）,提高群势25.15%;4个实验组间蜂蜜产量、封盖子数和蜂群群势都差异不显著（P> 0.05）。【结论】不同组配的蜜蜂信息素对中华蜜蜂生产及繁殖性能都有积极作用,为进一步在养蜂生产中推广蜜蜂信息素产品提供了理论支撑。     

中华蜜蜂Orco嗅觉受体基因的克隆、表达及亚细胞定位

【目的】克隆鉴定中华蜜蜂Apis cerana cerana的Orco嗅觉受体基因, 并对其在工蜂触角上进行免疫荧光定位。【方法】利用RT-PCR技术克隆中华蜜蜂Orco基因, 并对其编码的氨基酸序列进行生物信息学分析, 使用Real-time PCR技术鉴定其在中华蜜蜂不同发育时期及不同组织的表达谱; 利用免疫荧光定位技术在中华蜜蜂工蜂触角中对Orco进行亚细胞定位。【结果】获得中华蜜蜂Orco基因的全长cDNA序列, 命名为AcerOrco (GenBank登录号： JF968610.1), 其全长为1 434 bp, 编码477个氨基酸, 预测其含7个跨膜结构以及4个位于细胞膜外的亲水区。表达谱分析显示, AcerOrco在卵、 幼虫和蛹期呈低丰度表达, 1日龄及内勤蜂时期主要在触角和足中表达, 且在1日龄的触角中表达量最高; 采集蜂时期的触角、 头(去除触角)、 胸、 腹和翅中均有较高丰度的表达。亚细胞定位结果显示, AcerOrco不仅在采集蜂触角鞭节上大量表达（尤其在触角鞭节第1亚节中表达量较高）, 而且常成对出现, 并且发现AcerOrco可能主要在触角毛形感器的外部神经元(outer dendrite, OD)以及板形感器的树突神经元中表达。【结论】成功克隆了AcerOrco基因全长, 获得了其表达谱, 且将其定位于工蜂采集蜂的触角感器神经元上, 最终推测AcerOrco与中蜂嗅觉发育和触角感器功能密切相关。     

Organization and postembryonic development of glial cells in the visual system of the Chinese honeybee, Apis cerana cerana (Hymenoptera:Apidae )

神经胶质作为视觉系统的重要成分之一,对视觉系统的发育及功能起着重要的作用.本研究通过组织解剖观察、免疫组织化学等技术,对中华蜜蜂Apis cerana cerana幼虫和蛹的视觉系统中神经胶质的类型和发育过程进行了比较研究.研究表明:在中华蜜蜂视觉系统中,根据神经胶质的位置和形态主要分为表面神经胶质、皮层神经胶质和神经纤维网神经胶质3种类型;神经胶质主要来源于视柄和视叶中的神经胶质前体中心;神经胶质细胞数量的增加一方面来自于细胞的迁移,另一方面来自于神经胶质细胞自身的分裂增殖.本研究为昆虫神经胶质的发育以及功能研究提供理论基础.     

中华蜜蜂视觉系统中神经胶质的组成和胚后发育

神经胶质作为视觉系统的重要成分之一, 对视觉系统的发育及功能起着重要的作用。本研究通过组织解剖观察、 免疫组织化学等技术, 对中华蜜蜂Apis cerana cerana幼虫和蛹的视觉系统中神经胶质的类型和发育过程进行了比较研究。研究表明： 在中华蜜蜂视觉系统中, 根据神经胶质的位置和形态主要分为表面神经胶质、 皮层神经胶质和神经纤维网神经胶质3种类型; 神经胶质主要来源于视柄和视叶中的神经胶质前体中心; 神经胶质细胞数量的增加一方面来自于细胞的迁移, 另一方面来自于神经胶质细胞自身的分裂增殖。本研究为昆虫神经胶质的发育以及功能研究提供理论基础。     

The preferences of the honeybee (Apis mellifera) for different visual cues during the learning process

By working with very simple images, a number of different visual cues used by the honeybee have been described over the past decades. In most of the work, the bees had no control over the choice of the images, and it was not clear whether they learned the rewarded pattern or the difference between two images. Preferences were known to exist when untrained bees selected one pattern from a variety of them, but because the preferences of the bees were ignored, it was not possible to understand how natural images displaying several cues were detected. The preferences were also essential to make a computer model of the visual system. Therefore experiments were devised to show the order of preference for the known cues in the training situation. Freely flying bees were trained to discriminate between a rewarded target with one pattern on the left side and a different one on the right, versus a white or neutral target. This arrangement gave the bees a choice of what to learn. Tests showed that in some cases they learned two or three cues simultaneously; in other cases the bees learned one, or they preferred to avoid the unrewarded target. By testing with different combinations of patterns, it was possible to put the cues into an order of preference. Of the known cues, loosely or tightly attached to eye coordinates, a black or blue spot was the most preferred, followed by strong modulation caused by edges, the orientation of parallel bars, six equally spaced spokes, a clean white target, and then a square cross and a ring. A patch of blue colour was preferred to yellow.     

中华蜜蜂气味受体基因AcerOR58的序列与时空表达分析

[目的]通过分析中华蜜蜂Apis cerana cerana气味受体基因AcerOR58编码蛋白的理化性质、结构特征,明确AcerOR58时空表达特性,为该基因后续的功能研究奠定基础.[方法]利用多种生物信息学软件预测分析AcerOR58序列及其编码蛋白的结构特性,采用邻接法构建系统进化树.利用实时荧光定量PCR技术分析AcerOR58在不同发育阶段工蜂触角及采集蜂不同组织的表达差异.[结果]AcerOR58基因的开放阅读框(ORF)长1 230 bp,编码409个氨基酸,成熟蛋白分子量为47.147 ku,理论等电点8.46,无信号肽,含有6个跨膜结构且N端位于胞内,31个潜在的磷酸化位点,在第80-405位氨基酸之间存在一个昆虫气味受体家族7tm_6 superfamily保守结构域.AcerOR58与西方蜜蜂Apis mellifera的AmelOR58亲缘关系最近,核苷酸序列一致性高达96.67％,氨基酸序列一致性高达97.31％.AcerOR58在采集蜂(15-25日龄)阶段的表达量较高,且在触角中的表达量极显著高于其他组织(P＜0.01).[结论]AcerOR58具有昆虫气味受体的结构特征,该基因特异性高表达于中华蜜蜂采集蜂触角中,推测其功能与识别外界蜜粉源的花香气味物质有关.     

中华蜜蜂细胞色素CYP9E2基因克隆及其表达分析

【目的】克隆中华蜜蜂Apis cerana cerana细胞色素CYP9E2基因的完整编码区序列,分析CYP9E2基因在工蜂体内的表达特征,为研究该基因的生物学功能提供理论基础。【方法】以解剖获得的中华蜜蜂采集蜂中肠组织为材料,提取总RNA。利用RT-PCR技术克隆中华蜜蜂CYP9E2基因的编码区。采用多种生物信息学软件分析该基因的核苷酸和氨基酸序列,利用荧光定量PCR技术(quantitative real-time PCR)分析其在中华蜜蜂工蜂成虫期不同阶段(初生蜂、哺育蜂、守卫蜂以及采集蜂)头部和中肠组织中的相对表达量及在饲喂氟氯苯菊酯后工蜂中肠组织中的表达变化。【结果】克隆获得中华蜜蜂CYP9E2基因(命名为Ac CYP9E2)mRNA序列,长度为1 600 bp(Gen Bank登录号:KX394629),编码区长1 494 bp,编码497个氨基酸,其蛋白质分子量为57.026k D,等电点为8.32。系统发育树显示,中华蜜蜂Ac CYP9E2与西方蜜蜂Apis mellifera、小蜜蜂Apis florea CYP9E2基因聚成一支。对中华蜜蜂工蜂成虫期不同阶段头部和中肠组织Ac CYP9E2相对表达量测定发现,该基因在中华蜜蜂工蜂成虫期不同阶段的表达量存在一定差异,其中,采集蜂头部和中肠组织中Ac CYP9E2相对表达量均显著高于初生蜂、哺育蜂以及守卫蜂(P0.05),而且4个阶段工蜂中肠组织中的Ac CYP9E2相对表达量均显著高于其头部(P0.05)。饲喂氟氯苯菊酯后,工蜂中肠组织中Ac CYP9E2的相对表达量显著高于对照组(P0.05)。【结论】推测Ac CYP9E2可能参与了中华蜜蜂机体外源物质的代谢与解毒过程。     

中华蜜蜂卵表面微观结构及化学成分初步研究

为认识蜜蜂的工蜂监督机制,利用扫描电镜分别观察了中华蜜蜂Apis cerana cerana不同类型卵（蜂王产的受精卵、未受精卵以及工蜂产的未受精卵）的表面超微结构,同时用气 质联用技术测定了各类型卵新产卵（0 h）和5 h卵的表面化学信息素成分。结果表明：中华蜜蜂3种卵表面均覆有六边形的结构,结构内充满小突起,3种卵的大小及表面超微结构无显著差异。受精卵表面化学信息素种类比未受精卵更为丰富,C27∶2和C27∶1可能是卵带有的标记化学信息素。     

中华蜜蜂咽下神经节的结构和胚后发育

咽下神经节是昆虫腹神经索的第一个复合神经节, 主要调节口器附肢的活动。本研究通过形态解剖、 BrdU免疫组织化学等技术, 对中华蜜蜂Apis cerana cerana咽下神经节的组织结构和胚后发育过程进行了比较研究。结果表明： 中华蜜蜂的咽下神经节由上颚、 下颚和下唇3个神经节组成。在胚后发育过程中, 细胞增殖的活跃期主要集中在预蛹和蛹发育的第1天, 增殖活动一直持续到蛹发育的第4天结束。根据神经胶质细胞的位置和形态, 咽下神经节中的神经胶质可分为3种类型--表面神经胶质、 皮层神经胶质和神经纤维网神经胶质。本研究为蜜蜂神经系统的发育和功能研究提供了理论基础。