Expression of chemosensory proteins in hairs on wings of Locusta migratoria (Orthoptera: Acrididae)

The hairs on the wings of Locusta migratoria were observed and mapped using light microscopy, as well as by scanning and transmission electron microscopy. Based on their ultrastructure, we can distinguish four main types of hairs on the wings of adult L. migratoria , viz, short, medium and long hairs, and sensilla chaetica. The long hairs are located only on the ventral surface of the hindwing, whereas the other three types are present both on the dorsal and ventral surfaces of forewing and hindwing in both sexes. Medium hairs and sensilla chaetica are significantly more abundant on the dorsal surface of forewings in both females and males, than on the ventral surface, whereas the opposite was observed for short hairs (P < 0.01). No significant difference between males and females was observed in the density of any type of hairs (P > 0.1). Several dendritic branches, enveloped by a dendrite sheath, are situated in the lymph cavity of sensilla chaetica. Instead, no dendritic structure was observed in the cavity of the other three types of hairs. Immunocytochemical localization of chemosensory proteins (CSPs) was performed on ultrathin sections of hairs on wings. The antiserum against chemosensory proteins from L. migratoria (LmigCSP-II) strongly labelled sensilla chaetica, with gold granules only found in the outer sensillum lymph. In addition, the epidermal cell membrane of the wing was stained by the antiserum against LmigCSP-II. The other three types of hairs were never labelled. The results indicate that the wings might involve in contact chemoreception process.     

The chemosensilla on tarsi of Locusta migratoria (Orthoptera: Acrididae): Distribution,ultrastructure, expression of chemosensory proteins

The chemosensilla on the tarsi of Locusta migratoria were mapped using light microscopy, as well as scanning and transmission electron microscopy. Only chemosensilla chaetica were found on the tarsi. On the basis of their ultrastructure, these can be grouped into three main subtypes: short, long, and sunken sensilla chaetica. Short sensilla chaetica can be further divided into two groups containing 6 or 7 neurons. Long sensilla chaetica are mainly located on the lateral surface of the tarsi. Short sensilla chaetica were mainly found on the dorsal surface of the tarsi. Sunken sensilla chaetica were only found on the ventral surface, such as the pulvilli and arolium. Immunocytochemical localization of chemosensory protein (CSP) was performed on ultrathin sections of chemosensilla on tarsi. The antiserum against LmigCSP‐II intensively labeled all three types of sensilla chaetica. Gold granules were concentrated in the outer sensillum lymph surrounding the dendrite sheath, while the inner sensillum lymph containing dendrite branches was never labeled. Massive labeling with the anti‐LmigCSP‐II was also found in cuticle of the pulvilli on the ventral surface of tarsi. J. Morphol. 2009. © 2009 Wiley‐Liss, Inc.     

Chemosensory proteins of Locusta migratoria (Orthoptera: Acrididae)

We describe a family of proteins abundantly expressed in the chemoreception sensory organs, the antennae and legs, of the desert locust, Locusta migratoria (Orthoptera, Acrididae). Using polymerase chain reaction-based approaches and homology screenings, "OS-D"-like proteins were identified in L. migratoria. The different sub-types (LmigOS-Ds) are very similar to each other and share about 50-70% identity with OS-Ds from Drosophila melanogaster and Periplaneta americana. A similar degree of identity was also observed with moth OS-Ds. Northern blot analysis revealed a strong expression of the LmigOS-Ds in the antennae and legs, suggesting their involvement in chemosensory processes. Despite the lack of direct evidence for their role in chemosensation, LmigOS-Ds and their homologs seem to constitute a large protein family, characterized by a striking abundance and diversity among insect chemosensory organs.     

东亚飞蝗胚胎体节的形成过程

体节形成是昆虫胚胎发育过程中的关键问题.东亚飞蝗Locusta migratoria manilensis(Meyen)是一种重要的农业害虫,其体节形成的时序过程尚无详细报道.本研究采用免疫组化和品红染色方法研究了室内人工饲养东亚飞蝗的体节形成过程.结果表明:完成受精后,细胞核开始分裂并向卵表面迁移.细胞核到达卵表面的...     

短时高温暴露下东亚飞蝗雌虫血淋巴蛋白表达分析及质谱鉴定

为研究短时高温对东亚飞蝗Locusta migratoria manilensis Meyen血淋巴蛋白的影响,采用Bradford法、SDSPAGE电泳和质谱等方法,对东亚飞蝗雌虫血淋巴样品进行检测。结果表明:短时高温对血淋巴蛋白含量有显著影响(P0.01),36℃-42℃范围内,随温度升高,血淋巴蛋白浓度亦升高,其中39℃、42℃处理组与对照组差异显著(P0.01);短时高温对血淋巴蛋白种类存在一定影响,对照组雌虫血淋巴中存在11种蛋白,高温处理后,4种蛋白含量逐渐增加,6种蛋白含量没有明显变化,1种蛋白消失;经质谱检测,鉴定了5种蛋白,分别为载脂蛋白前体、酚氧化酶原、2个储存蛋白和19 kDa血淋巴蛋白,另外6条蛋白未被鉴定。推测载脂蛋白前体、酚氧化酶原、储存蛋白在东亚飞蝗应对高温胁迫过程中发挥重要作用。     

Contact chemoreceptors among wind-sensitive head hairs of Locusta migratoria L. (Orthoptera: Acrididae)

The peg sensillum, a type of sensillum intermingled with the long hair sensilla in the hair fields on the head of Locusta migratoria (Orthoptera: Acrididae), was studied by light, transmission and scanning electron microscopy. The peg sensilla have the features typical of contact chemoreceptors; each peg is innervated by 5 sense cells; and 4 of the dendrites, enclosed within a “dendritic canal”, pass through the central lumen of the peg to the distal part, below the apical pore. The 5th dendrite ends in a tubular body at the caudal side of the peg articulation. Each distal segment of the 5 dendrites has a ciliary structure (9 ×2+0) at the transition to the short proximal segment, which in each case turns to the side to merge with the soma of the bipolar sense cell. Four sheath cells are associated with the group of sense cells and they are surrounded by a 5th, special epidermis cell. The innermost (thecogen) sheath cell (No. 1) encloses the receptor-lymph space 1 and forms the cuticular sheath; sheath cells 2 and 4 form the boundary of the large volume of receptorlymph space 2. The number of sheath cells is discussed with reference to other insect sensilla and in a phylogenetic context.     

Occurrence and genetics of black‐eyed migratory locusts,Locusta migratoria (Orthoptera: Acrididae)

Black‐eyed Locusta migratoria appeared in albino locusts as a result of crossing between a short‐winged strain originating from Tsushima Island, Japan, and an albino strain originating from Okinawa Island. The black eye trait was recessive to the white eye trait because the crosses between black‐ and white‐eyed albino locusts produced only individuals with white eyes in the F₁ generation. In the F₂ generation, black‐ and white‐eyed individuals appeared in a ratio of 1:3, indicating that the black eye trait was controlled by a simple Mendelian unit. The black eye trait showed no genetic association with other traits including wing morph, adult body dimensions and classical morphometric ratios such as hind femur length / head width and forewing length / hind femur length.     

Evidence for widespread genomic methylation in the migratory locust, Locusta migratoria (Orthoptera: Acrididae)

The importance of DNA methylation in mammalian and plant systems is well established. In recent years there has been renewed interest in DNA methylation in insects. Accumulating evidence, both from mammals and insects, points towards an emerging role for DNA methylation in the regulation of phenotypic plasticity. The migratory locust (Locusta migratoria) is a model organism for the study of phenotypic plasticity. Despite this, there is little information available about the degree to which the genome is methylated in this species and genes encoding methylation machinery have not been previously identified. We therefore undertook an initial investigation to establish the presence of a functional DNA methylation system in L. migratoria. We found that the migratory locust possesses genes that putatively encode methylation machinery (DNA methyltransferases and a methyl-binding domain protein) and exhibits genomic methylation, some of which appears to be localised to repetitive regions of the genome. We have also identified a distinct group of genes within the L. migratoria genome that appear to have been historically methylated and show some possible functional differentiation. These results will facilitate more detailed research into the functional significance of DNA methylation in locusts.     

The functions of the maxillary palps of Acrididae (Orthoptera)

The functions of the maxillary palps of acridids have been investigated in experiments with Locusta migratoria migratorioides (R. & F.), Schistocerca gregaria (Forsk.) and Chorthippus parallelus (Zett.). The palp makes small irregular flicking movements and more extensive rapid vibrations described as palpation. Both activities involve the same basic movements of the palp segments. Palpation is essentially related to feeding and in normally feeding insects the palps play an important role in food selection, but as starvation is prolonged they become unimportant. Selection depends on appropriate chemical stimulation from the cuticle on the surface of the leaf. Palpation on this leads to head lowering and to biting if appropriate tactile stimuli are then received.The control of biting is discussed. The frequent but very brief, contacts of the sensilla on the tips of the palps with the substratum during palpation may enable them to relay information to the central nervous system more or less continuously without adapting.

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Zusammenfassung Die Funktionen der Maxillartaster von Locusta migratoria migratorioides (R. & F.), Schistocerca gregaria (Forskål) und Chortippus parallelus (Zett.) wurden unter Anwendung verschiedener Verfahren untersucht. Die Tasterbewegungen wurden kinematographisch analysiert und als Schlagen und Tasten klassifiziert; das letztere tritt hauptsächlich kurz vor und nach, aber ebenso gut auch während der Nahrungsaufnahme auf. Die Sensillen an den Tasterenden berühren beim Tasten häufig das Substrat.Vorausgesetzt, daß die Insekten nicht gehungert hatten, wurde Bellis allein nach Betasten abgelehnt; nach längerem Hungern erfolgte jedoch die Ablehnung erst nach Beißen. Weitere Versuche, bei denen Extrakte der Kutikula von Poa oder Bellis verwendet wurden, zeigten, daß Annahme oder Ablehnung von der chemischen Natur des Substrats abhängen, welches von den Tastern berührt wird. Betasten eines geeigneten Substrats führte nicht unmittelbar zum Beißen, ruft aber eine Verhaltensweise hervor, die ihrerseits das Beißen fördert.Daraus wird geschlossen, daß die von den Tastern wahrgenommenen chemischen Reize für die Nahrungswahl der Acrididen wichtig sind, wenn die Insekten normal fressen. Nach Hunger ruft mechanische Reizung der Taster oder anderer Mundteile Beißen auch beim Fehlen geeigneter chemischer Reize hervor. Die häufigen kurzen Kontakte der Palpen mit dem Substrat während des Tastens erlauben offenbar den Sensillen, einen beständigen Strom von Informationen an das Zentralnervensystem zu liefern, ohne selbst angepaßt (ermüdet) zu werden.

     

Response of Locusta migratoria tibetensis Chen (Orthoptera: Acrididae) to climate warming over the Tibetan plateau

As a region with one of the most important terrestrial ecosystems, the Tibetan plateau is both sensitive to and vulnerable to climate warming. Locusta migratoria tibetensis Chen, an endemic species on the Tibetan plateau, is likely to be affected by climatic warming. In our studies, accumulated degree‐days (ADD) of L. Migratoria T.C. were calculated based on data from 90 weather stations over the Tibetan plateau from 1961 to 2005. Trend lines show that across weather stations, ADD increased at a rate of 1.17 DD/a during climate warming. The majority of weather stations (82.2%) showing increases in ADD were located towards the west of the Tibetan plateau. At higher elevations, the increase in the ratio of annual ADD to the station mean ADD was higher relative to those at lower elevations. A linear regression model between ADD and geographical position was established to create raster maps of ADD in ArcGIS 9.2. The area of potential locust distribution (APD) was estimated to be 42 420 km², mostly distributed along major rivers on the Tibetan plateau. In warmer years, the APD increased sharply over study periods. A new area of potential distribution would appear in the north Tibetan plateau if the climate warming continued. In the south‐east Tibetan plateau, the locust would expand its range northwards or westwards along the river valleys, and the locust APD would also rise in elevation.     

The annual cycle of Saharan populations of Locusta migratoria cinerascens (Orthoptera: Acrididae: Oedipodinae) in Algeria

Until recently, the Algerian populations of Locusta migratoria cinerascens were considered as sedentary and solitarious. In the Central Sahara and bordering areas, the extension of pivot-irrigated areas since 1980, has resulted in the creation of biotopes favourable to the reproduction of this acridid, particularly in the cereal crops of the Adrar and Ouargla regions. From analysis of the archives of the National Institute of plant Protection, and our personal work carried out from 1995 to 2004, we can provide new information on the life cycle of L. m. cinerascens. We show that, in the cereal crops of the regions studied, there are three generations of this acridid per annum: the first one developing on corn in the spring, until the harvest; the second a spring-summer generation in market gardens in the vicinity of wheat stubble, or in early-growth Sorghum crops, and a third, summer-autumn generation, that develops on summer cereal crops and probably over-winters as diapausing adults until the following February.     

Development of the suboesophageal body during embryogenesis without diapause in Locusta migratoria (linnaeus) (Orthoptera: Acrididae)

The development and the ultrastructural changes of the suboesophageal body were studied during embryogenesis of Locusta migratoria (Orthoptera : Acrididae). The suboesophageal body develops from the mandibular coelomic cavities. It differentiates early, before the completion of germ band segmentation (stage IIIc), the other mesodermal cells remaining undifferentiated. The cells of the suboesophageal body rapidly develop a structure similar to that of nephrocytes. They consist of a peripheral transfer zone and a perinuclear zone, the site of synthesis and storage. Material absorbed by endocytosis is taken up by α-vacuoles, then stored in β-vacuoles. Golgi vesicles, tubules and vesicle complexes may be involved in the secretory activity of the cells. The activity of the suboesophageal body is maximal until stage VI, after katatrepsis, after which degeneration begins. Very few cells remain at eclosion and they are completely degenerated. The suboesophageal body may be involved in the regulation of embryonic haemolymph composition, and it develops according to its function. The suboesophageal body differentiates early and is thus functional when the haemolymph first forms in the subgerminative space. It degenerates after the differentiation of the pericardial cells and the fat body, which regulate haemolymph composition.     

On the pronymphal stage in the migratory locust (Locusta migratoria, Orthoptera, Acrididae)

The structure of the integument, somatic and visceral muscles, midgut, and Malpighian tubules were investigated at the late stages of the embryonic and early postembryonic development of the migratory locust, Locusta migratoria, to assess the organization of its pronymphal stage. In its morphogenetic features, the vermiform locust larva sometimes called the pronymph corresponds to the first nymphal instar covered with the second embryonic cuticle which has not been shed. Since the first-instar locust nymphs before and after the shedding of this embryonic cuticle differ significantly in many morphological characters, two consecutive phases of this nymphal instar can be distinguished: the first phase existing from the moment of development of the third embryonic cuticle to the shedding of the second one; the second phase existing from the shedding of the second embryonic cuticle to the formation of the cuticle of the second nymphal instar. Since the pronymphal stage should precede the nymph stage, it may be concluded that the pronymph of the locust is fully embryonized and covered with the second embryonic cuticle, which is also typical of other insects with hemimetabolous development (Konopová and Zrzavý, 2005). Therefore, it would be erroneous to refer to the vermiform first-instar nymph as the pronymph, because the two stages are separated by molting and formation of a new cuticle.     

Selection of Antonospora locustae (Protozoa: Microsporidae) with higher virulence against Locusta migratoria manilensis (Orthoptera:Acrididae)

Antonospora locustae is a microsporidian parasite of grasshopper insects that is used as a biological control agent. We report on laboratory selection of isolates from different regions with increased virulence. Bioassays were conducted against third instar nymphs of Locusta migratoria manilensis. AL2008L01 was originally imported from the USA in 1986, AL2008M01 was isolated from Melanoplus differentialis in USA and AL2008F01 was isolated from infected Fruhstorferiola tonkinensi collected in Guangdong, China. The results showed that all three isolates can infect the locust and that pathogenicity increased gradually with increased dose. The LD₅₀ values of the original isolates at the highest dose (5×10⁶ spores/nymph) were 19, 23 and 22 days and LD₅₀ values were 3.2×10⁵, 3.4×10⁶ and 0.7×10⁶ spores/g, respectively. After selecting for three generations, the virulence of all isolates increased significantly. LT_50s were reduced to 17, 20 and 21 days at the highest dose (5×10⁶ spores/nymph) and LD_50s were reduced to 1.4×10⁵, 2.5×10⁵ and 1.7×10⁵ spores/g.     

Effect of Paranosema (Nosema) locustae (Microsporidia) on morphological phase transformation of Locusta migratoria manilensis (Orthoptera: Acrididae)

Field and laboratory studies demonstrated that Paranosema (Nosema) locustae had significant effects on the morphological phase transformation of Locusta migratoria manilensis (Meyen 1835). In the field, spraying P. locustae on gregarious locusts caused a substantial population reduction by 16 days after treatment, with most of the surviving locusts being phase solitaria. However, the effects of P. locustae on locust phase transformation began before direct mortality had caused a substantial reduction in locust density: locust numbers were still high at day 10, but locusts had already transformed to phase transiens. Laboratory assays showed that while a low dose of P. locustae had no effect on phase transformation, at a higher dose of 1×10⁵ spores/mL, locusts had F/C ratios that were significantly (P<0.05) more solitaria than untreated locusts, with locusts having ratios that were either phase solitaria or on the solitaria side of phase transiens. In a second laboratory experiment that analysed the effects of locust density on phase transformation by P. locustae, there was no obvious effect of density on female locusts 10 days later as all were solitaria at all locust densities. At day 16, female locusts were transiens at higher densities, but were solitaria at 4/cage. With males there were lesser effects. These results provide new evidence for P. locustae having sub-lethal effects on locust phase transformation at a wide range of locust densities.     

飞蝗可溶型海藻糖酶基因的序列分析及mRNA表达特性

海藻糖酶是海藻糖代谢过程中的一个关键酶, 在昆虫发育和能量调节中具有重要作用, 为进一步探讨海藻糖酶基因的功能, 本文分析了飞蝗Locusta migratoria一可溶型海藻糖酶基因的氨基酸序列并对其mRNA表达特性进行了研究。结果表明： 该海藻糖酶（TRE, GenBank登录号： FJ795020）不含有跨膜结构。系统进化树分析结果显示, 该酶与大豆蚜Aphis glycines、 豌豆蚜Acyrthosiphon pisum、 褐飞虱Nilaparvata lugens和灰飞虱Laodelphax striatella可溶型海藻糖酶具有较近的亲缘关系, 因此我们将该酶的基因命名为LmTre-1。对该基因在不同组织和发育时期表达量的荧光定量 PCR 分析表明： LmTre-1在卵发育前期、 中期的表达量都很低, 卵发育后期表达量显著提高; LmTre-1在5龄若虫和成虫被检测的组织部位中均有表达, 在体壁中的表达量最高, 其次是在脂肪体、 肌肉、 气管、 精巢及卵巢中; 5龄飞蝗刚蜕皮后LmTre-1在体壁中的表达量较高, 随着生长发育其表达量逐渐降低; LmTre-1在成虫发育期体壁中稳定高表达。LmTre-1的mRNA表达特性与几丁质合成酶1基因非常相似, 据此推测该基因可能与体壁几丁质的合成相关。本研究为深入探讨该基因的生理功能提供了重要的基础数据, 并为以海藻糖酶为杀虫靶标的农药筛选奠定实验基础。     

Differences in numbers of sensilla on the antennae of solitarious and gregarious Locusta migratoria L. (Orthoptera: Acrididae)

It has been assumed that differences in the behaviour of solitarious and gregarious locusts arise from differences in the central nervous processing of information, and that no essential differences occur in the sensory systems. This investigation reveals that solitarious adults and fifth instar nymphs of Locusta migratoria (Orthoptera: Acrididae) have more olfactory sensilla on the antennae than comparable gregarious phase insects. Behavioural differences could thus arise as a result of different peripheral inputs. It is suggested that social facilitation, resulting from the gregarious habit, permits a decrease in individual sensitivity with an associated reduction in the numbers of olfactory sensilla. The numbers of contact chemoreceptors are proportional to the lengths of the antennal annuli and do not differ greatly between the phases.     

Effect of Paranosema locustae (Microsporidia) on the behavioural phases of Locusta migratoria (Orthoptera: Acrididae) in the laboratory

The ability of parasites to modify the behaviour of their hosts is a wide spread phenomenon, but the effects of microsporidian parasites on locust behaviour remain unexplored. Here the frequencies of directional changes (ND) and jumping (NJ) per minute of gregarious locusts infected with 2000 spores of the microsporidian parasite Paranosema locustae were significantly different from those of untreated locusts 10 and 16 days after infection, being similar to values for solitary nymphs. In contrast, the behaviour of locusts inoculated with the lower doses of 200 spores/locust was sometimes like that of solitary nymphs. At other times, behaviour was intermediate between solitary and gregarious, i.e. transitional. The rearing density did not affect the turning and jumping behaviour of infected locusts, and their behaviours were similar to those of solitary locusts at 10–16 days after infection. Our study demonstrates that infection with P. locustae may lead gregarious locusts to change some of their behaviour to that typical of solitary locusts.