A miniaturized assay to quantify effects of chemicals or physical stimuli upon locust activity

Solitary and gregarious locusts differ in many traits, such as body color, morphometrics and behavior. With respect to behavior, solitary animals shun each other, while gregarious animals seek each other＇s company, hence their crowding behavior. General activity, depending on the temperature, occurs throughout the day but is much lower in solitary locusts. Solitary locusts occasionally fly by night while gregarious locusts fly regularly during daytime （swarming）. In search of new assays to identify substances that control or modify aspects of （phase） behavior, we designed a simple activity assay, meant to complement existing behavioral measurement tools. The general activity is reflected in the number of wall hits, that is, the number of contacts between the locust and the vertical walls of a small arena. Using this single parameter we were able to quantify differences in total activity of both nymphs and adults of isolation-reared （solitary）, regrouped- and crowdreared （gregarious） locusts under different conditions. Furthermore, we demonstrate that there are inter- and intra-phase dependent differences in activities of 5th instar nymphs afar injections of the three different adipokinetic hormones.     

飞蝗变型及体色多型的内分泌控制机理

飞蝗具变型现象,散居型和群居型在形态特征、生理机能、行为及体色等方面存在明显差异。保幼激素已被证实能诱导飞蝗散居型绿色的出现。近年,从飞蝗心侧体成功地分离了一种神经肽——黑化诱导激素([His^7]-corazonin),并用白化型进行检测,证实了其对体色黑化作用的活性。不同时间对飞蝗若虫注射不同剂量的[His^7]-corazonin,能诱导出现除绿色以外的散居型体色,如浅褐色、褐色、赤褐色、黑色等;也能诱导出现群居型的体色,即黑色配以橘黄色的底色。而且,对散居型若虫注射[His^7]-corazonin能诱导其形态向群居型转换。这些研究证实[His^7]-corazonin对飞蝗的变型有着重要的控制作用,但又不是唯一的。     

东亚飞蝗行为和形态型变的判定指标

通过田间、室外罩笼、室内行为测试等一系列实验,研究了东亚飞蝗群居型和散居型之间的行为和形态差异。确立了东亚飞蝗不同生态型个体的形态和行为指标．结果表明,雌雄散居型蝗蝻每分钟的跳跃次数均在1．4以下,转向次数分别在1．3和1．4以下;雌雄群居型蝗蝻每分钟的跳跃次数均在1．6以上,转向次数分别在1．6和1．5以上．群居型蝗虫的跳跃次数、转向次数显著高于散居型蝗虫。所以跳跃次数、转向次数可作为东亚飞蝗行为型变判定指标．在同型不同性别的蝗虫之间行为型变指标没有显著差异．F／C值可作为4龄以上东亚飞蝗的形态型变判定指标。而E／F值可作为东亚飞蝗成虫的形态型变判定指标．两型的F／C值都随龄期的增长而增加,且同龄期雄虫F／C均大于雌虫F／C．F／C、E／F值在不同型态和同型不同性别间均存在极显著差异．因此,确定两型形态型变标准时应将雌、雄虫分开,即雌性和雄性散居型第4、5龄及成虫的F／C值分别大于2．5、2．8、3．3和2．6、2．9、3．5;雌性和雄性群居型第4、5龄及成虫的F／C值分别小于2．5、2．7、3．1和2．5、2．8、3．3．成虫的E／F值也可以作为成蝗形态型变的判断指标．     

Detection of DNA strand breakage in a marine amphipod by agarose gel electrophoresis: exposure to X-rays and copper

This article describes the leading steps to develop an assay of DNA damage for the marine amphipod Gammarus locusta, using agarose gel electrophoresis (AGE). To test the sensitivity and feasibility of the AGE technique, X-ray assays were performed with naked DNA and with live amphipods. These positive controls demonstrated the effectiveness of the AGE technique to not only discriminate distinct levels of DNA strand breakage in a dose-dependent manner, but also to identify and quantify the type of strand breakage induced. It was also shown that it is possible to detect DNA damage using whole-body DNA extracts from amphipods. To explore the potential of this technique for use in ecotoxicological studies with amphipods, a 96-h waterborne-copper toxicity test was performed. Copper-induced DNA strand breakage was first observed after 24 h of exposure, and was recorded again at 96 h, at a copper concentration of 20 μg l -1 . The absence of strand breakage after 48 h of exposure is discussed in the light of the underlying mechanisms of copper toxicity and DNA repair. These studies demonstrated the feasibility of including DNA damage as a biomarker in ecotoxicological studies with amphipods. Information gained from the use of this biomarker would help with the interpretation of chronic toxicity tests and would contribute to our understanding of the impact of genotoxic insult in marine invertebrates, particularly crustaceans.     

氟虫脲对东亚飞蝗和中华稻蝗几丁质合成酶基因表达的影响

为了探讨氟虫脲可能的作用靶标及毒性机制, 本研究以重要农业害虫东亚飞蝗Locusta migratoria manilensis (Meyen)和中华稻蝗Oxya chinensis (Thunberg)为材料, 采用简并引物扩增中华稻蝗几丁质合成酶1基因（OcCHS1）的部分cDNA序列; 以氟虫脲浸渍法处理2龄中期中华稻蝗及1, 2和3龄东亚飞蝗若虫为处理组, 丙酮处理为对照组, 使用RT-PCR和实时荧光定量PCR方法分析氟虫脲对蝗虫几丁质合成酶基因mRNA表达的影响。结果获得的OcCHS1部分cDNA序列, 其长度为312 bp, 编码104个氨基酸, GenBank登录号为HM214491, 与东亚飞蝗几丁质合成酶1基因（LmCHS1）在氨基酸水平上相似度达95％。RT-PCR结果显示, 处理组几丁质合成酶1扩增带均强于对照组。实时荧光定量PCR结果表明: 与对照组相比, 处理组中华稻蝗2龄中期若虫OcCHS1 mRNA表达提高了1.02倍, 东亚飞蝗1, 2, 3龄若虫LmCHS1 mRNA表达分别提高了34%, 82%和89%, 差异显著（P<0.05）。分析基因表达提高的原因是几丁质合成受阻后基因表达水平的一种代偿性增加, 由此推测几丁质合成酶可能是氟虫脲作用的靶标之一。     

基于光投影数字化传感系统的飞蝗表面三维结构

本文基于结构光投影数字化传感系统构建东亚飞蝗Locusta migratoria manilensis表面的三维结构。首先得到其外形尺寸和肢体各部分拓扑结构,然后从被调制的条纹结构光中恢复出物体表面高度变化的相位信息,再通过相位和高度的对应关系重建出物体表面三维特征。此结果将为农药研制和施用设计提供科学依据,对促进蝗虫的防治以及虚拟昆虫研究起到积极作用。     

Genetic divergence among geographical populations of the migratory locust in China

The migratory locust, Locusta migratoria L., which is distributed widely in the East Hemisphere, has long been regarded as the most important agricultural pest and a model insect. Its distribution ranges from thetropic and the subtropical zone, to the temperate zone, and to the cold-temperate zone. Because of the wide geographical distribution and adaptations, many geo- graphical populations of the migratory locust dis- played apparent variation in morphology, life history, physiology, and ot…     

Selective actions of Lynx proteins on different nicotinic acetylcholine receptors in the locust,Locusta migratoria manilensis

Nicotinic acetylcholine receptors (nAChRs) are major neurotransmitter receptors and targets of neonicotinoid insecticides in the insect nervous system. The full function of nAChRs is often dependent on associated proteins, such as chaperones, regulators and modulators. Here, three Lynx (Ly‐6/neurotoxin) proteins, Loc‐lynx1, Loc‐lynx2 and Loc‐lynx3, were identified in the locust, Locusta migratoria manilensis. Co‐expression with Lynx resulted in a dramatic increase in agonist‐evoked macroscopic currents on nAChRs Locα1/β2 and Locα2/β2 in Xenopus oocytes, but no changes in agonist sensitivity. Loc‐lynx1 and Loc‐lynx3 only modulated nAChRs Locα1/β2 while Loc‐lynx2 modulated Locα2/β2 specifically. Meanwhile, Loc‐lynx1 induced a more significant increase in currents evoked by imidacloprid and epibatidine than Loc‐lynx3, and the effects of Loc‐lynx1 on imidacloprid and epibatidine were significantly higher than those on acetylcholine. Among three lynx proteins, only Loc‐lynx1 significantly increased [³H]epibatidine binding on Locα1/β2. The results indicated that Loc‐lynx1 had different modulation patterns in nAChRs compared to Loc‐lynx2 and Loc‐lynx3. Taken together, these findings indicated that three Lynx proteins were nAChR modulators and had selective activities in different nAChRs. Lynx proteins might display their selectivities from three aspects: nAChR subtypes, various agonists and different modulation patterns.

     

Ageing adipokinetic cells in Locusta migratoria: an ultrastructural morphometric study

Summary A morphometric study was made of the ultrastructure of adipokinetic cells in resting adults of Locusta migratoria at 3, 23, and 43 days after imaginal ecdysis. The nucleus, rough endoplasmic reticulum, and Golgi apparatus enlarge with age, which indicates that the synthesis and packaging of secretory substances increases during ageing. The size of the storage compartment, consisting of secretory and ergastoplasmic granules, does not increase earlier than 23–43 days after imaginal ecdysis. The lysosomal compartment markedly enlarges between 3 and 23 days; later on, the growth of this compartment, especially of autophagosomes, is less prominent. This suggests that lysosomal destruction initially compensates for the production of new secretory granules, assuming that exocytosis of secretory granules by adipokinetic cells is insignificant in resting locusts. Afterwards, lysosomal destruction may no longer be sufficient to prevent over-production of secretory granules, as is suggested by the increase in the number of these granules between 23 and 43 days. This coincides with the appearance of a considerable number of large ergastoplasmic granules, which represent a spatially more efficient form of storage of secretory material than the much smaller secretory granules. The increase with age in the amount of secretory products indicates that the biosynthetic activity of the adipokinetic cells is not (finely) tuned to their releasing activity.     

The endocrine control of phase transition: some new aspects

Abstract. The present article summarizes some recent findings relating to the underlying mechanism of phase transition in locusts, from the nonswarming solitarious phase to the swarming gregarious phase. These phases differ in many traits, such as colouration, morphometrics and behaviour. The most comprehensive theory at present to explain the switch from the nonswarming to the swarming form is that the locusts are brought together by the heterogeneity of the environment. They gather at preferential structures and food plants and physical contact then stimulates individuals to gregarize. Phase change can also be transferred across generations by maternal pheromones. The endocrine regulation of phase polymorphism is still not fully understood. The role of ecdysteroids has been studied, so far with no final conclusion. It is remarkable that the prothoracic glands persist longer in isolated-reared adults, which implies that these glands continue to play a role, although they no longer release important amounts of ecdysteroids. Juvenile Hormone, without any doubt, induces certain solitarious characteristics, such as green colouration, but is not the primary causal factor. A real breakthrough was the discovery of [His⁷]-corazonin, made possible by using a novel assay system, the Okinawa albino mutant of Locusta migratoria , which was known to be deficient in this hormone. This peptide, which is produced in the brain and is most likely released via the corpora cardiaca, promotes the gregarious black pigmentation. It also plays a role in morphometrical phase change as well as in behavioural alterations. Corazonin is apparently quite an important peptide not only in locusts, but also in insects in general.