Involvement of ecdysone in the control of meiotic reinitiation in oocytes of Locusta migratoria (Insecta,orthoptera)

Following their biosynthesis in the follicle cells of vitellogenic ovaries, large amounts of ecdysteroids pass into the oocytes where they accumulate and persist during ovulation and egg-laying. The present paper shows that free ecdysone is unevenly distributed in the oocytes exhibiting the highest concentrations in the region of the posterior pole where the final sequences of nuclear maturation, including germinal vesicle breakdown (GVBD), occur. A correlative study indicates that the concentrations of free ecdysone in this region are particularly high (10 to 20 μM) during two periods of meiotic reinitiation observed in the oocytes: reinitiation I, leading from prophase I to metaphase I with GVBD; and reinitiation II, from metaphase I to the end of meiosis. In vitro incubations of oocytes in meiotic arrest (prophase I) in the presence of exogenous ecdysone demonstrate that complete reinitiation (including GVBD) can be triggered in a dose-dependent manner by this hormone.     

Localization of serotonin-like immunoreactivity in the eyestalk of the prawn Palaemon serratus (Crustacea, Decapoda, Natantia)

A serotonin-like substance in the organ of Bellonci in the eyestalks of embryos, larvae, and adults of the prawn Palaemon serratus was visualized by the use of two specific antisera against serotonin (5-hydroxytryptamine; 5-HT) in combination with peroxidase-antiperoxidase (PAP). The organ of Bellonci, characterized by compact onion bodies distally and degenerating onion bodies proximally, was the only site of the serotonin-like substance in adults, as well as during development in embryos and larvae. Variations in the content of the 5-HT analogue in the adult were detected during the molting cycle. There was more immunoreactivity in specimens fixed at night than in those fixed in daytime. Likewise, colchicine and nialamide injections enhanced the immunoreactivity of the serotonin-like substance. Extirpations of the medulla externa X organ (MEX), a neurosecretory cell group of the optic ganglion medulla externa, produced the same effect.     

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...     

气候变暖背景下新疆和中哈边境亚洲飞蝗适生区预测及变化

亚洲飞蝗Locusta migratoria migratoria是中国新疆与哈萨克斯坦边境区域重要害虫之一,具迁飞性且繁殖能力强,对农牧业经济造成巨大伤害。本文以亚洲飞蝗为研究对象,根据其发生区和相关生物学特性,利用CLIMEX 4.0.2和ArcGIS 10.2相结合,预测其在新疆和中哈边境区域亚洲飞蝗的潜在地理分布。亚洲飞蝗的适生区在新疆境内集中分布在北疆(42.69～48.29°N),包括伊犁、博尔塔拉中部、塔城西北部及南部、阿勒泰西部、昌吉和乌鲁木齐南部地区;哈萨克斯坦境内主要分布在北纬47.03～51.65°N区域,包括东哈萨克斯坦州北部、阿拉木图州东南部境内的湖泊及河流沿岸地区,这与亚洲飞蝗的现有分布点和历史发生记录吻合。未来气候变暖情景下,亚洲飞蝗在中哈边境新疆境内的适生区逐渐扩大,其中在阿勒泰、伊犁、昌吉及乌鲁木齐的高度适生区面积增加,哈密开始出现高度适生区;在哈萨克斯坦边境区域的适生区略有增加,其中东哈萨克斯坦州地区高度适生区面积减少明显。研究结果对区域内亚洲飞蝗的预测预报和综合防控具有重要意义。     

Optimization of parental RNAi conditions for hunchback gene in Locusta migratoria manilensis (Meyen)

An investigation on the optimization of parental RNA interference (RNAi) conditionsfor hunchback {hb) gene in Locusta migratoria manilensis (Meyen) was conducted.Double stranded RNA (dsRNA) corresponding to hb gene was injected into haemocoel offemale adults of L.migratoria manilensis.Embryos developed from the eggs laid by theinjected adults on the 7th day after eclosion showed observable effects of RNAi for hb.Thesilencing effect after delivery treatment of dsRNA for hb gene was maintained for moretha...     

白洋淀东亚飞蝗持续大发生浅析

进入 90年代以来 ,白洋淀蝗区东亚飞蝗Locustamigratoriamanilensis(Meyen)持续大发生 4次 ,是50年来十分罕见的。究其原因 ,主要受白洋淀水位变化的影响 ,其中 8月水位的高低直接影响翌年夏蝗发生程度 ,其次是生态环境和气候条件等因素的影响。     

Development of the tympanal organ in larvae of the migratory locust (Locusta migratoria)

Summary The tympanal organ of the migratory locust acquires its definitive form during larval development. All the receptor cells (90–100) are present in the 1st instar, whereas the differentiation of the tympanum and the cuticular structures it bears proceeds in steps from one instar to the next. The elevated process is the earliest such structure to appear (2nd instar); it is followed by the pyriform vesicle (3rd instar) and folded body (4th instar). The styliform body first appears in the imago. Although the typical arrangement of the receptor cells is already discernible in the 1st instar, some of the attachment sites change during development, the final configuration appearing only in the imago.Supported by the Deutsche Forschungsgemeinschaft (Ka 498/2)     

Characterization of histones and chromatin of the hepatopancreas in Palaemon serratus (Crustacea Natantia)]

The chromatin of shrimp hepatopancreas has been extracted from isolated nuclei and characterized. Nuclei were prepared in the presence of Cu++ and phenyl methyl sulfonyl fluoride in order to inhibit the nuclease and protease activities throughout the different purification steps. The purified nuclei are heterogenous in size and show a density of 1,367 g/ml determined on saccharose - glucose gradients. After washing in 0,14 M NaCl and then in 10(-2) M Tris-HCL, pH = 7,6, the nuclei were disrupted in water. The solubilized chromatin was precipitated in 0,15 M.NaCl. This chromatin is characterized by a high level of RNA (RNA/DNA = 0,38) and of non histone proteins (NHP/DNA = 0,6). The denaturation curve showed only one Tm at 69 degrees in 2.10(-4) M.EDTA. When the chromatin was extracted in the presence of staphylococcal nuclease, the Tm reached 80 degrees C. The kinetics of the digestion by the staphylococcal nuclease have been studied and show that 10 per cent of hydrolysis occurs within the first minute. The repeat length of DNA as determined with the polymers of higher order is 189 +/- 5 base pairs. The existence of nucleosomes was confirmed by electron microscopy. The superstructure of chromatin was not completely destroyed after solubilisation with a Potter. The histones were studied by gel electrophoresis after differential staining. The most important feature consists in the presence of two H1, two H2A and two H4. The acetylation levels of the histones were followed after injection of 14C-acetate in vivo. The subfraction H1, 0 was acetylated. Only one H3 was present and the two H2A fractions showed the same level of acetylation. H2B migrated faster than the H2A fractions like in Echinoderms. The two H4 fractions corresponded to two differently acetylated forms. Shrimp hepatopancreas histones were fractionated by molecular sieving on Biogel P 100 and characterized according to their electrophoretic properties as well as their amino-acid content. The amino-acid compositions of the different histone fractions were nearer to Echinoderm and Sipunculid histones, than Calf thymus homologue histones. All the fractions show a weaker basicity. The H3 fraction was the only one showing a lesser variability when compared to Calf thymus H3. The non histone proteins were extracted in 10(-2) M Tris-HCL, pH = 8 and 0.1 per cent SDS. A series of 50 proteins was detected. 80 per cent of the total amount of protein was localized in a molecular weight range comprised between 40 000 and 80 000 daltons. These proteins were compared to the histones and total proteins of sonicated chromatin solubilized by SDS in order to detect proteasic effects.     

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.     

Influence of soil moisture on egg cold hardiness in the migratory locust Locusta migratoria (Orthoptera: Acridiidae)

Abstract.  The present study investigates the influence of environmental moisture on cold hardiness of the migratory locust, Locusta migratoria . The water content of locust eggs kept in soil at 30 °C varies according to the moisture content of the substrate. In turn, it can significantly affect the supercooling point of locust eggs (range from −26 to −14.8 °C) and the mortality when exposed to subzero temperatures. Environmental moisture influences the supercooling capacity of eggs and their survival at low temperature. When locust eggs of the same water content are exposed to subzero temperatures under different soil moistures, their mortality varies between short-time exposure and long-time exposure at subzero temperatures. Given a short-time exposure, mortality in wet soil is lower than in dry soil due to the buffering effect of soil water against temperature change. The pattern of egg mortality is reversed after long-time exposure at low temperature, suggesting that inoculative freezing may be an important mortality factor. It is suggested that interactions between soil moisture and low temperature can influence the cold hardiness of locust eggs, and partial dehydration is beneficial to over-wintering eggs of the migratory locust.     

Fecal volatile components elicit aggregation in the oriental migratory locust,Locusta migratoria manilensis (Orthoptera: Acrididae)

Abstract The aggregation components from fecal volatiles of the oriental migratory locust, Locusta migratoria manilensis were identified with gas chromatographic/electroantennographic detector (GC‐EAD), GC‐MS (mass spectrometry) analyses and behavioral bioassays. Both last instar nymphs and adults of the oriental migratory locust have similar aggregation pheromones in their volatiles. A total of 11 electrophysiologically active compounds, namely, hexanal, cyclohexanol, heptanal, phenol, 2,5‐dimethyl‐pyrazine, benzyl alcohol, benzaldehyde, guaiacol, nonanal, 2,6,6‐trimethyl‐2‐cyclohexene‐1,4‐dione and decyl aldehyde were identified in the fecal volatiles of 2‐day‐old immature adult male locusts. Only hexanal, nonanal, benzaldehyde, cyclohexanol and 2,5‐dimethyl‐pyrazine elicited significant aggregation responses in immature 2‐day‐old adult males. However, adult males had significantly lower behavioral responses to synthetics of five single compounds than the blend of cyclohexanol, 2,5‐dimethyl‐pyrazine, benzaldehyde, nonanal, hexanal in ratios of 100: 100: 2: 60: 30 in the range of 30–60 μg/mL. We propose that it is the blend of these five compounds that plays a key role in eliciting and sustaining aggregation in gregarious oriental migratory locusts. These results also showed that the aggregation pheromones of the oriental migratory locust are significantly different from those found in the desert locust.     

A comparison of dietary selection behaviour in larval Locusta migratoria and Spodoptera littoralis

ABSTRACT. . Final instar nymphs of the oligophagous acridid Locusta migratoria (L.) and larvae of the polyphagous noctuid Spodoptera littoralis (Boisduval) were fed for 4, 8 or 12 h, the conditioning period, on one of four artificial diets. Of these, diet PC contained 20% protein and 10% digestible carbohydrate; another, P, contained 20% protein but with the digestible carbohydrate component replaced by cellulose; a third, C, had the protein component substituted by cellulose, and the fourth, O, had both protein and digestible carbohydrate replaced. After this conditioning period, insects were given a choice of two diets, P and C, and hence an opportunity to select for the nutrients, if any, which were lacking in their previous food. Amounts eaten and selection behaviour were then recorded in detail for a total of 9 h. This paper deals with total amounts of diet eaten during the conditioning and choice periods. Spodoptera larvae were more sensitive than the locusts to being fed a nutritionally inadequate conditioning diet, and ate only small quantities of the P, C and O diets as compared with the PC diet, irrespective of the duration of conditioning. Locusts, on the other hand, when restricted to the P diet continued to eat relatively large amounts of it throughout a 12 h conditioning period. Those nymphs fed the C diet ingested large quantities (more than of the PC diet) up until 8 h, after which intake fell. When offered a choice, both species selected for the nutrients missing from the conditioning diet, even if the conditioning period had been as short as 4 h. During the first hour of choice locusts selected the P diet if they had been previously fed C and the C diet if previously fed P. Those deprived of both nutrients increased consumption of both P and C diets. Spodoptera larvae were more sensitive to prior deprivation of digestible carbohydrate than of protein. During the first hour of choice they selected the C diet if previously fed P or O but did not choose the P diet if previously fed C. In the subsequent 8 h of choice, however, a strong selection for the P diet after previous deprivation became apparent. In the locust, the selection for nutrients missing from the conditioning diet continued for the following 8 h of choice but became masked by a tendency, shown by all nymphs, to select C over P. The functional significance and possible physiological basis of all these responses is discussed.     

东亚飞蝗中肠几丁质酶基因的克隆、序列分析及组织定位

通过RACE方法,克隆了东亚飞蝗Locusta migratoria manilensis (Meyen)几丁质酶基因 (LmChi)cDNA全序列 (GenBank 登录号：EF092841)。获得的cDNA全长1 604 bp,其中可读框1 452 bp, 编码483个氨基酸。推测其氨基酸序列与18家族昆虫几丁质酶有较高的相似性。与其他几丁质酶一样,东亚飞蝗几丁质酶序列也包含一个信号肽、一个几丁质酶活性位点、一个碳端丝氨酸富集区和一个几丁质结合域。半定量RT-PCR研究表明,LmChi基因只在东亚飞蝗不同发育阶段的中肠组织中表达,而在东亚飞蝗体壁、前肠和后肠均没有发现LmChi基因的转录。     

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。     

Muscle activity underlying sexual behaviour in male locusts (Locusta migratoria) after normal and hormonally disturbed development

Abstract. . The activities of three sets of muscles, representing the major components of the abdominal muscle system, were recorded in the pregenital abdominal segments of adult males of Locusta migratoria. The muscular activities were associated with three consecutive actions of the male's mating behaviour: abdominal lowering, S-bending and hooking-up. These three actions depend mainly on the relative coordination of the dorsal versus ventral longitudinal muscles; the activity of the dorsoventral muscles is rather unpredictable, except in the sixth and seventh abdominal segments where they are regularly involved in hooking-up. Studies on male sexual behaviour-linked muscle activity in precocene-induced fifth-instar adultiforms and azadirachtin-induced fifth-instar over-aged nymphs revealed no distinct differences between these creatures and normal mature males. Therefore the muscle activities associated with these adult-specific behavioural acts can be released precociously, i.e. their occurrence is not dependent on a distinct series of instars. Moreover, the results obtained with over-aged nymphs demonstrate that morphogenesis and ethogenesis (i.e. formation of neural circuits controlling adult-specific behaviour) are not closely dependent on each other.     

Flight fuel related differences between solitary and gregarious locusts (Locusta migratoria migratorioides)

Abstract. Flight fuel relations of crowded and isolated Locusta migratoria migratorioides were investigated in younger (12–16 days after fledging) and older (27–30 or 27–32 days after fledging) adult males.No phase polymorphism dependent differences were found in resting haemolymph carbohydrate levels of the younger locusts.In the older age group, resting haemolymph carbohydrate levels were slightly though significantly higher in the isolated than in the crowded locusts.Injection of various doses of synthetic adipokinetic hormones (AKHs) did not induce marked changes in haemolymph carbohydrate levels and no differences were found between crowded and isolated locusts.A 30 min flight led to the same decrease in haemolymph carbohydrate levels of isolated and crowded locusts, 43.3% and 44.6% of the resting levels, respectively.We concluded, therefore, that the results do not seem to indicate that isolated locusts rely more heavily on carbohydrates as flight fuel than crowded locusts.Hyperlipaemic responses to flight were less intense in isolated than in crowded locusts, but phase polymorphism dependent differences in flight-induced increase of haemolymph lipid levels were not parallel in 12–16-day-old and 27–32-day-old males.In the younger age group the difference was mainly in the duration of flight needed to induce full response which appeared already after 20 min of flight in the crowded locusts, but only after 45 or 60 min of flight in the isolated ones.In contrast, the older isolated locusts showed markedly lower haemolymph lipid elevations than the crowded locusts even after 30, 45 or 60 min of flight.The hypothesis is forwarded that isolated locusts have a rather coarse adipokinetic strategy focused on a single long-distance migratory flight, whereas gregarious locusts possess a fine adipokinetic balance for reiterative migratory flights and saving fuel reserves for unpredictable long-distance migrations.     

Mechanisms of organophosphate resistance in a field population of oriental migratory locust, Locusta migratoria manilensis (Meyen)

The susceptibilities to three organophosphate (OP) insecticides (malathion, chlorpyrifos, and phoxim), responses to three metabolic synergists [triphenyl phosphate (TPP), piperonyl butoxide (PBO), and diethyl maleate (DEM)], activities of major detoxification enzymes [general esterases (ESTs), glutathione S-transferases (GSTs), and cytochrome P450 monooxygenases (P450s)], and sensitivity of the target enzyme acetylcholinesterase (AChE) were compared between a laboratory-susceptible strain (LS) and a field-resistant population (FR) of the oriental migratory locust, Locusta migratoria manilensis (Meyen). The FR was significantly resistant to malathion (57.5-fold), but marginally resistant to chlorpyrifos (5.4) and phoxim (2.9). The malathion resistance of the FR was significantly diminished by TPP (synergism ratio: 16.2) and DEM (3.3), but was unchanged by PBO. In contrast, none of these synergists significantly affected the toxicity of malathion in the LS. Biochemical studies indicated that EST and GST activities in the FR were 2.1- to 3.2-fold and 1.2- to 2.0-fold, respectively, higher than those in the LS, but there was no significant difference in P450 activity between the LS and FR. Furthermore, AChE from the FR showed 4.0-fold higher activity but was 3.2-, 2.2-, and 1.1-fold less sensitive to inhibition by malaoxon, chlorpyrifos-oxon, and phoxim, respectively, than that from the LS. All these results clearly indicated that the observed malathion resistance in the FR was conferred by multiple mechanisms, including increased detoxification by ESTs and GSTs, and increased activity and reduced sensitivity of AChE to OP inhibition.     

Further experimental evidence for the involvement of ecdysone in the control of meiotic reinitiation in oocytes of Locusta migratoria (Insecta, Orthoptera)

Ecdysone has recently been shown to be able to trigger meiotic reinitiation in vitro in submature oocytes of Locusta. In the present study we have experimentally depressed (by 60-70%) ecdysone biosynthesis in the ovaries of adult females by rearing them on a diet with a modified sterol profile. Mature oocytes from such females fail to undergo normal reinitiation, but when incubated in vitro, can be induced to break their meiotic arrest by the addition of exogenous ecdysone. These results lend further support to the hypothesis that in Locusta, ovarian ecdysone is involved in the control of meiotic reinitiation.     

Chill injury in the eggs of the migratory locust, Locusta migratoria （Orthoptera： Acrididae）： the time-temperature relationship with high-temperature interruption

Abstract Mortality of the overwintering egg of the migratory locust, Locusta migratoria L. , was attributed to chill injury because of its occurrence well above the egg's super cooling point. In this study, two parameters, upper limit of chill injury zone (ULCIZ) and sum of the injurious temperature (SIT), were used to examine the locust egg's cold hardiness. The value of ULCIZ for the locust egg is 1.06 ± 0.54°C, and the SIT is -329.7 (hour · degree). The superoxide dismutase (SOD) and catalase (CAT) activities changed dramatically after cold stress, indicating that oxygen and hydroxide free radicals are probably efficiently detoxified at low temperatures. It was suggested that the nature of chill injury in locust egg might be a complex of metabolic disorder and a non-proportional decrease in enzymatic reaction and transports, because the LDH activity at low temperature increased significantly and the ATPase activity decreased with prolonged duration of exposure to low temperatures. The results from high temperature interruption revealed that the high temperature intervals significantly increased the survival of locust eggs.