金龟子绿僵菌中性海藻糖酶基因的克隆及其表达特性分析

根据中性海藻糖酶NTL基因的同源序列设计引物,PCR扩增出杀蝗专一菌株———金龟子绿僵菌CQMa102NTL基因片段,利用5′_RACE和3′_RACE扩增出NTLcDNA的5′和3′端序列,经拼接得到CQMa102NTL基因cDNA全长。根据其全长cDNA序列,设计引物PCR扩增出CQMa102NTL的完整基因。为了解该基因的上游调控信息,采用PanhandlePolymeraseChainReactionAmplification方法扩增其上游序列。序列分析表明,CQMa102NTL全长DNA3484bp,cDNA全长2385bp,编码737个氨基酸的蛋白,推测蛋白分子量为83.1kD;含有3个内含子,包含一个依赖于cAMP的磷酸化作用位点(RRGS)和一个钙附着位点(DTDGNMQITIED);上游序列含有一个压力反应元件(CCCCT);与金龟子绿僵菌广谱性菌株ME1NTL的核苷酸序列和氨基酸序列分别具有93%和99%同源性,由此确定该序列为金龟子绿僵菌中性海藻糖酶基因序列。Southern杂交表明,NTL基因在CQMa102基因组中为单拷贝。Northern杂交表明,NTL基因转录出约2.5kb的mRNA单带,在液体培养条件下,对数生长前期表达水平最高,对数生长后期降到最低,进入稳定生长期后表达水平又有所提高。金龟子绿僵菌CQMa102中性海藻糖酶基因DNA全长和cDNA全长登录GenBank,登录号分别为:AY557613,AY557612。     

Chemo-discriminatory neurones in the sub-oesophageal ganglion of Locusta migratoria

The sub-oesophageal ganglion of Locusta migratoria was searched for neurones responsive to stimulation of the maxillary palps using intracellular recording techniques. Two plant stimuli were used: wheat, a host plant and cabbage, an unacceptable non-host plant. The stimuli were presented to the palp as both intact leaf tissue and as droplets of aqueous solutions of plant extracts. All the sampled neurones that responded to stimulation of the palp also responded to simple mechanical stimulation. However, 25% of the neurones exhibited consistent differences in response to the two plants when presented as both leaf tissue and droplets, strongly suggesting that they also received a chemosensory input. These differential responses most commonly took the form of differences in the duration of cell activity and/or variation in the latency of the onset of response. The receptive fields of differentially responding neurones were confined to the maxillary palp, or at most to the maxillary and labial palps.     

Purification of Choline Acetyltransferase from the Locust Schistocerca gregaria and Production of Serum Antibodies to This Enzyme

Choline acetyltransferase (ChAT; EC 2.3.1.6) was purified from the heads of Schistocerca gregaria to a final specific activity of 1.61 mumol acetylcholine (ACh) formed min-1 mg-1 protein. The molecular mass of the enzyme as determined by gel filtration is 66,800 daltons. The final enzyme preparation showed one major band at 65,000 daltons on sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis, which corresponds with the native molecular mass of the enzyme, a band at 56,000 daltons, and two bands at 40,500 and 38,000 daltons. Antibodies raised against ChAT in rabbit react only with the active band on native gel after Western blotting. They strongly react with the 65,000-dalton polypeptide band on Western blots of SDS gel separation of pure preparation of enzyme and with both the 65,000- and 56,000-dalton bands after SDS gel separation of crude extract.     

Mechanical characterization of the key portions in locust semi-lunar processes under different strain rates

The excellent rapid jumping and kicking of locusts are largely attributed to the power amplification mechanisms due to the semi-lunar processes (SLP) at their distal metathoracic femurs, especially dorsal-core (i.e., portion II) and ventral-core parts (i.e., portion III). The physiological range of strain rates at the two portions of locust SLP is quite broad in the periods of energy storage and release (approximately three orders). However, it still remains elusive how the mechanical properties of the two SLP portions change with the strain rate. We identified the elastic moduli and material compositions of SLP portions II and III by using nanoindentation and confocal laser scanning microscope. Apparent and creep-corrected reduced elastic moduli were calculated to represent the total energy absorption and storage, respectively. The results revealed that both portions II and III exhibit strain rate-sensitive elastic moduli, regardless of water content. The efficiency of elastic energy storage is only 51–70% in the case of low strain rate. This work can deepen our understanding in the energy storage and release mechanisms in locust locomotion and further provide guidelines for biomimetic design of power amplification apparatus in jumping robots.     

In search for non-steroidogenic functions of the prothoracic glands of the desert locust, Schistocerca gregaria: a peptidomic and proteomic approach

The only well established function of the prothoracic glands (PGs) of insects is the production of ecdysteroids. In gregarious locusts, like in most insect species, the PGs degenerate soon after the adult molt. In this way they resemble the thymus of mammals, a gland with an important role in the build up of the immune system in young animals. In adult solitarious locusts the PGs persist much longer, however without producing substantial amounts of ecdysteroids. In the literature the existence of a well developed rough endoplasmic reticulum and Golgi complex system has been repeatedly reported, suggesting an active role in peptide or/and protein synthesis and release. The nature of the secreted products remained unknown. Our pepdidomic analysis of an acidified methanolic extract of PGs of last instar gregarious nymphs did not yield any indication for the presence of known locust or other peptides. The peptide release assay was also negative. For our proteomic analysis, we developed an EST-based identification strategy. We successfully identified 50 protein spots on a two dimensional map. In addition to typical protein synthesis-related proteins, a number of proteins with a role in detoxification processes were found, suggesting some role of the PGs in the defense system.     

Behavioural phase change in the Australian plague locust, Chortoicetes terminifera, is triggered by tactile stimulation of the antennae

Density-dependent phase polyphenism is a defining characteristic of the paraphyletic group of acridid grasshoppers known as locusts. The cues and mechanisms associated with crowding that induce behavioural gregarization are best understood in the desert locust, Schistocerca gregaria, and involve a combination of sensory inputs from the head (visual and olfactory) and mechanostimulation of the hind legs, acting via a transient increase in serotonin in the thoracic ganglia. Since behavioural gregarization has apparently arisen independently multiple times within the Acrididae, the important question arises as to whether the same mechanisms have been recruited each time. Here we explored the roles of visual, olfactory and tactile stimulation in the induction of behavioural gregarization in the Australian plague locust, Chortoicetes terminifera. We show that the primary gregarizing input is tactile stimulation of the antennae, with no evidence for an effect of visual and olfactory stimulation or tactile stimulation of the hind legs. Our results show that convergent behavioural responses to crowding have evolved employing different sites of sensory input in the Australian plague locust and the desert locust.     

Windborne displacements of Desert Locusts from Africa to the Caribbean and South America

The Desert Locust is a major pest of agriculture in Africa, the Middle East and South-West Asia and swarms are known to make downwind flights over hundreds and thousands of kilometres between seasonal breeding areas. At the end of summer in 1988, swarms of locusts were moving north and south along the western margins of North Africa and in October and November, swarms crossed the Atlantic Ocean and invaded the Caribbean and neighbouring parts of South America for the first recorded time. Because of the extent of the migration and the evolutionary significance of linkages between Old and New World species of locusts, the weather associated with the migrations was studied and trajectory analysis was used to identify the source areas and estimate the flight times. Locusts were moving offshore from western North Africa throughout the autumn and on three occasions migrated west of 40° W with easterly Trade winds. Two trans-Atlantic crossings coincided with the passage of easterly waves. Over 100 trajectories were constructed at 950 and 850 hPa and within the time limit used ( 144-h), 28% successfully linked source and receptor areas. Minimum trajectory duration was 93-h, which is one-and-a-half times longer than the previously longest flight duration, derived for a similar migration to the British Isles in 1954. Upwind trajectories from the arrival areas, identified sources between 27 and 6° N in Africa, with most end-points located in Mauritania, Senegal, Gambia and Guinea-Bissau. Interspersed with the Atlantic crossings were a northward movement of locusts and an incursion of Saharan dust into Europe within the circulations of frontal depressions. While offshore migrations from northern Africa are common in autumn, the immigrants in the Caribbean and South America were probably at the extreme limits of flight endurance for the species. The results tend to confirm earlier hypotheses that New World species of locusts may have evolved from ancestral migrants from Africa.