东北地区亚洲飞蝗染色体核型分析

采用常规压片法对吉林省亚洲飞蝗Locusta migratoria migratoria(L.)的染色体核型进行分析.研究结果表明:亚洲飞蝗性别决定机制为X0型,染色体数目为2n♂=23,染色体组式4L+4M+3S+X,全部为端部着丝粒染色体,NF=23.染色体中最长(L1)与最短(S11)染色体之比大于4:1,臂比大...     

东亚飞蝗谷胱甘肽S-转移酶分离纯化

通过硫酸铵沉淀技术和GSH-agarose亲和层析对东亚飞蝗Locusta migratoria manilensis(Meyen)5龄若虫谷胱甘肽S-转移酶(glutathione S-transferases,GSTs)进行了分离纯化。结果表明GSTs活性在硫酸铵各沉淀段均有分布,但在55%～100%沉淀段活性较高,在硫酸铵饱和度为85%时比活力最高,达到420.33μmol/min/mg protein,纯化倍数为18.86。根据硫酸铵粗沉淀谷胱甘肽S-转移酶结果,选择硫酸铵浓度为60%～90%沉淀段进行GSH-agarose亲和层析,纯化后比活力最高达到1365.29μmol/min/mg protein,纯化倍数达到61.25。经SDS-PAGE鉴定,得到的GST为1条带,亚基的分子量约为24kDa。     

亚洲飞蝗发声器结构与鸣声时域特征研究

本文对亚洲飞蝗Locusta migratoria migratoria(L.)发声器的结构及鸣声的时域特征进行了研究.亚洲飞蝗主要的发声方式为前翅中闰脉-后足股节型,即前翅中闰脉的发声齿与后足股节内侧隆线相互摩擦发出鸣声.应用扫描电子显微镜技术对发声器的结构进行观察,结果表明,亚洲飞蝗前翅中闰脉具有单排规则排列的发声...     

GABA-like immunoreactivity of identified interneurons in the flight system of the locust,Locusta migratoria

Summary The transmitter content of identified inhibitory interneurons in the flight system of the locust, Locusta migratoria, has been characterized using antibodies raised against protein-conjugated gamma aminobutyric acid. Identified flight neurons were filled with the fluorescent dye, Lucifer Yellow. Serial sections of dye-filled neurons were incubated with an antibody to gamma aminobutyric acid which was subsequently tagged with a fluorescent marker. Excitatory motoneurons to wing muscles and 13 flight interneurons (3 excitatory, 7 inhibitory, and 3 with unknown synaptic effect) were examined. Neither the moto-neurons nor any of the 3 excitatory interneurons contained immunoreactive material. Six of the 7 inhibitory interneurons did contain immunoreactive material. All the neurons which contained immunoreactive material and whose synaptic effect is known were inhibitory. We conclude that most of the inhibitory flight interneurons which have been described use gamma aminobutyric acid as their transmitter. Interestingly, at least 1 set of interneurons known to be inhibitory does not use gamma aminobutyric acid. We predict that the 2 interneurons which do contain immunoreactive material and whose synaptic effect is not yet known will be found to have inhibitory roles in the operation of the flight circuitry.     

Patterns of respiration in Locusta migratoria nymphs when feeding

Flow‐through respirometry was used to investigate patterns of respiration of fifth‐instar Locusta migratoria L. nymphs fed a chemically defined, synthetic food. Each animal was recorded for up to 2.7 h, during which they had access to food and water ad libitum, and at least one meal was taken. The start of feeding was coincident with a sudden and rapid rise in respiration. Both carbon dioxide (CO₂) production and oxygen (O₂) consumption rose, the traces for the two gasses showing a high degree of alignment. The end of a meal correlated with a sudden and rapid decrease in respiratory rate towards resting levels. When feeding was interrupted by an intra‐meal pause, respiratory rate tended to drop marginally and then stabilize, before rising rapidly upon the resumption of feeding within the meal. Maximal rates of respiration during feeding represented a 3–4‐fold increase over those at rest. Walking and climbing within the chamber were not associated with any noticeable change in respiratory rate above baseline. When locusts were quiescent between feeding episodes, respiration was steady and continuous, rather than discontinuous. Possible causes for large changes in respiration during feeding are discussed.     

西藏飞蝗（Locusta migratoria tibetensis Chen）耐寒性理化指标

西藏飞蝗是青藏高原上的一种重要害虫.近年来, 西藏飞蝗在西藏、青海玉树以及川西高原上危害十分严重.通过对该虫各发育阶段过冷却能力测定,研究了各发育阶段虫体内生理生化物质与过冷却能力之间的关系,揭示了该虫耐寒性规律.结果表明,西藏飞蝗蝗卵的过冷却点和结冰点为最低,分别为-22.02℃和-16.36℃.在1～4龄蝗蝻期,过冷却点随体内水分、糖原和脂肪含量随着虫体的发育而升高,其过冷却点分别为-9.28℃、-8.51℃、-7.18℃、-6.46℃;到5龄蝗蝻时,虫体脂肪和糖原含量达最高值,过冷却点下降为-8.07℃.讨论了虫体脂肪、糖原和甘油含量变化与其耐寒力之间的关系.     

东亚飞蝗血细胞形态学研究

通过Wright's染色和光学显微镜对不同地区东亚飞蝗的血细胞进行了观察,发现东亚飞蝗Locusta migratoria manilensis(Meyen)的血细胞包括原血胞、浆血胞、粒血胞和类绛血胞4种类型.原血胞的细胞核为红色,而其它3种血细胞的细胞核均被Wright's染液染成红色和蓝色2种类型.故认为浆血胞是...     

东亚飞蝗主要过敏原的分析、鉴定与纯化

通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)分离东亚飞蝗Locusta migratoria manilensis(Meyen)的蛋白质组分并测定其分子量,收集过敏病人血清,采用免疫印迹(Western—blotting)法鉴定其过敏原成分,通过凝胶过滤层析对东亚飞蝗过敏原进行分离纯化。结果表明:东亚飞蝗蛋白粗提液条带大概有30条左右,其中主带大约有10条,相对分子量约为13、15、25、28、40、45、55、70、100、110ku,其中蛋白含量最丰富的约在70ku左右。免疫印迹结果显示,蝗虫过敏条带主要有5条,相对分子量分别约为19、29、38、70、130ku。通过凝胶过滤层析对东亚飞蝗过敏原进行分离纯化,得到了一个高纯度相对分子质量约为70ku东亚飞蝗过敏原,并且发现了一个相对分子质量约为130ku的蝗虫新过敏原。本研究为临床上蝗虫食物变态反应性疾病的诊断和治疗奠定基础。     

西藏飞蝗各发育阶段的耐寒性

西藏飞蝗Locusta migratoria tibetensis Chen是青藏高原的重要农牧业害虫。对该虫各发育阶段的过冷却点和结冰点的测定表明,西藏飞蝗卵的过冷却点和结冰点为最低,分别为-22.02℃、-16.36℃,4龄蝻的过冷却点和结冰点最高,分别是-6.46℃和-5.05℃,西藏飞蝗在甘孜以卵越冬。     

Spontaneous and neurally evoked contractions of visceral muscles in the oviduct of Locusta migratoria

The oviducts of Locusta migratoria are innervated by a pair of nerves which arise from, the seventh abdominal ganglion. A distinctive network of striated muscle fibres occurs in the oviducts. The lateral oviducts and common oviduct consist of an inner circular layer of muscle and an outer longitudinal layer of muscle. At the junction of the lateral and common oviduct an additional thin longitudinal layer is found adjacent to the basement epithelium. The oviducts contracted spontaneously when isolated from the central nervous system. These myogenic contractions took the form of peristaltic contractions in the lateral oviduct, and intermittent phasic-like contractions of the posterior regions of the lateral oviduct and the common oviduct. These phasic-like contractions were associated with individual complex potentials recorded extracellularly from the muscle fibres. In locusts that had been interrupted in the process of egg laying, there were large-amplitude action potentials, firing in a bursting pattern, in the oviducal nerves. These large action potentials were absent in locusts that had not been egg-laying. These action potentials were associated with both bioelectric potentials and mechanical events in the posterior region of the lateral oviduct and the common oviduct. Electrical stimulation of the oviducal nerve mimicked the effects of spontaneous action potentials, resulting in the appearance of monophasic potentials and contractions. The contractions were graded and dependent upon both frequency and duration of stimulation. It is concluded that the oviducts of Locusta are both myogenic and neurogenic. The role of these contractions in oviposition is discussed.     

白洋淀蝗区东亚飞蝗的分布与土壤的关系研究

从土壤的质地、含水量、pH值和含盐量等方面 ,研究了白洋淀东亚飞蝗Locustamigratoriamanilensis(Meyen)蝗区的土壤状况 ;并结合实地考察的样点处蝗虫密度情况 ,研究了蝗虫不同密度区土壤之间的差异。结果表明 ,研究区蝗虫密度在 3 0头 m2 以上的地区 ,土壤质地为粉砂壤土 ,土壤pH值为7 2 6～ 8 1 2 ,土壤含盐量为 0 0 67%～ 0 2 0 7%。粉砂壤土中 ,砂粒含量较多、粗粉粒含量较少的地方 ,是东亚飞蝗比较理想的生存与活动场所 ;土壤含水量偏高、呈弱碱性的地方 ,适于东亚飞蝗的生存和活动。在研究区 ,土壤含盐量差异对东亚飞蝗的密度分布没有明显的影响。     

不同食料植物对东亚飞蝗肠道细菌状况的影响

目的比较在相同的环境条件下不同食料植物对东亚飞蝗肠道细菌状况的影响。方法以墨西哥玉米、野生马唐为食料植物饲喂东亚飞蝗,在其成虫肠道内和粪沙中分离纯化细菌,获得22株菌株,分别对其培养性状、菌体形态、染色反应和生理生化性状进行系统研究。结果上述22个菌株分别属于稀有杆菌属（Rarobact-er）、短状杆菌属（Brachybacterium）、棒杆菌属（Corynebacterium）、棍状菌属（Clavibacter）、葡萄球菌属（Staphylococ-cus）、丙酸杆菌属（Propionibacterium）、埃希菌属（Eschrichia）、气微杆菌属（Aercmicrobium）、沙雷菌属（Serratia）、克雷伯菌属（Klebsiella）、沙门菌属（Salmonella）、志贺菌属（Shigella）、口腔球菌属（Stomatococcus）、短杆菌属（Brevibacteri-um）、地杆菌属（Terrbacter）、微小杆菌属（Exiguobacterium）、皮杆菌属（Dermabacter）、短小杆菌属（Curtobacterium）、纤维单胞菌属（Cellulomonas）、微杆菌属（Microbacterium）、乳杆菌属（Lactobacillus）和凝结芽胞杆菌属（Bacilluscoag-ulans）。结论不同食料植物对东亚飞蝗肠道细菌种类和数量有很大的影响。     

Effects of diet on the chemical composition of migratory locusts (Locusta migratoria)

An experiment was conducted to determine the effects of diet on the chemical composition of migratory locusts (Locusta migratoria L.). Fresh and dry weight and the contents of dry matter, ash, lipid, protein, Ca, K, Mg, Na, P, Cu, Fe, Zn, retinol, lutein, zeaxanthine, cryptoxanthin, carotenes, lycopene and gross energy were determined in penultimate instar and adult locusts, that had been fed three different diets. The locusts received a diet of grass or grass+wheat bran or grass+wheat bran+carrots. Adding wheat bran decreased the protein content and increased fat content (633 vs. 583 and 182 vs. 231 g/kg DM, respectively). Addition of carrots to the diet increased fat content further from 231 to 271 g/kg DM. Mineral concentrations of Ca, K, Mg, and Na, were significantly affected by diet. P, K, Cu, and Fe concentrations were significantly different in penultimate migratory locusts compared with adults. Wheat bran decreased the α-carotene content, which did not change by incorporating carrots in the diet. However, carrots did result in higher β-carotene concentrations. Retinol concentrations were increased by incorporating both wheat bran and carrots in the diet compared with the diet containing only grass. This study shows that the chemical composition of migratory locusts can be manipulated through the diet. As such, it enables nutritionists to adapt the chemical composition of live feeder insects to better meet the nutritional demands of predators.     

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。     

Structural and catalytic role of two conserved tyrosines in Delta-class glutathione S-transferase from Locusta migratoria

Glutathione S-transferases (GSTs) are an important family of detoxifying enzymes and play a key role in pesticide resistance in the insect. Tyrosine is essential for its detoxification function. In the present study, two conserved tyrosine residues are located at positions 108 and 116 in H-site of LmGSTD1. To elucidate how the two residues participate in the catalytic process and keeping structural stability, four mutants, Y108A, Y108E, Y116A, and Y116E, were generated. It was found that the four mutants affected the specific activity of LmGSTD1 in various degrees, depending on the types of substrate and reaction mechanism. Steady-state kinetics assay revealed that Y108E and Y116E had a significant influence on GSH-binding ability, which indicates the two tyrosine residues of H-site contribute to topology rearrangement of G-site. Both Y116A and Y116E exhibited lower CDNB-binding affinity, suggesting that Y116 takes part in hydrophobic substrate binding. The thermostability assay, intrinsic, and 8-anilino-1-naphthalenesulfonic acid (ANS) florescence results showed that the two tyrosine residues were involved in regulation of active-site conformation. Finally, homology modeling provided evidence that the two tyrosines in H-site participate in hydrophobic substrate binding. Furthermore, Y108 is closer to the S atom of S-hexylglutathione. In conclusion, the two tyrosines in LmGSTD1 are important residues in both the catalytic process and protein stability.     

Characterization of juvenile hormone-binding proteins of hemolymph of Locusta migratoria

The binding of juvenile hormone (JH) by components from hemolymph of adult female Locusta migratoria was characterized to establish whether hemolymph JH-binding proteins could be distinguished from a protein of fat body (BP-1) that may be a JH receptor. Hemolymph was analyzed by the hydroxyapatite assay, gel separation chromatography, polyacrylamide gel electrophoresis, and density gradient centrifugation. Three fractions that bound JH were separated from whole hemolymph by DEAE cellulose column chromatography, and these differed from all three cytosol-binding components. The major hemolymph component (H-A) showed relatively stable binding of JH, a slight loss of binding capacity after delipidation, and a K_d for JH-I of 16 nM. The K_ds for JH-l and JH-lll with unfractionated hemolymph were 26 and 42 nM respectively. The order of effectiveness of competitors for binding of [³H]JH-l was JH-lll > JH-l ? methoprene > hydroprene ? acids of methoprene and hydroprene. The data indicated that unlabeled JH-lll was bound more effectively than its radioactive counterpart. The sedimentation values determined by sucrose density gradient ultracentrifugation were 13-14 S for hemolymph, and the sedimentation value was not altered by the inclusion of 0.4 M KCl throughout the gradient. The data indicated that H-A resembled the specific JH carriers and differed from the putative receptor of fat body cytosol by several criteria.     

溴氰菊酯对飞蝗羧酸酯酶基因表达的影响

【目的】研究溴氰菊酯作用下飞蝗羧酸酯酶基因的mRNA表达特性,为溴氰菊酯的代谢解毒及飞蝗Locusta migratoria防治中抗性风险的评估提供基础资料。【方法】本文采用不同剂量溴氰菊酯处理3龄飞蝗,提取总RNA,体外反转录合成cDNA模板,采用Real-time PCR技术分析飞蝗羧酸酯酶基因在溴氰菊酯不同浓度和不同时间处理后的表达模式。【结果】飞蝗经不同浓度溴氰菊酯处理12 h后,LmCesA3和LmCesE1表现为诱导效应;除LmCesA2外,其余羧酸酯酶基因经溴氰菊酯LD30剂量处理后分别在不同的时间点表现为诱导效应。【结论】5个羧酸酯酶基因LmCesA1、LmCesA3、LmCesD1、LmCesE1和LmCesI1可以被溴氰菊酯诱导,表明其可能参与飞蝗对溴氰菊酯的代谢解毒及抗性产生。     

西藏飞蝗的生物学特性

西藏飞蝗Locusta.migratoria tibetensis Chen在四川甘孜州1年发生1代,某些地方(乡城县)1年发生不完整的2代,即以卵越冬,翌年3月下旬开始孵化出土,4月中、下旬为孵化盛期,1~3龄始盛期为4月中旬~5月中旬,高峰期为5月下旬,7月上旬初始羽化,7月下旬~8月上旬为羽化盛期,8月上旬始见产卵,8月下旬~9月上旬为产卵盛期,第1代成虫较早产下的卵块在条件适宜的情况下可于当年9月上旬孵化出土,但孵化出的蝗蝻不能越冬。该虫卵、全蝻期及全世代的发育起点温度分别为14.2,16.1,14.6℃,有效积温为179.1日.度、360.0日.度、787.8日.度。在18,21,24,27和30℃等5种恒温条件下其平均世代历期214.4,133.3,79.2,66.3和50.7d。     

东亚飞蝗羧酸酯酶基因的克隆和原核表达

羧酸酯酶是昆虫体内重要的代谢解毒酶系,其主要功能是水解和结合内源性和外源性含有酯键的有毒物质,减缓其到达靶标部位的时间。东亚飞蝗Locusta migratoria manilensis(Meyen)是我国重要的农业害虫,对其羧酸酯酶基因克隆和表达有助于深入探索杀虫剂代谢毒理机制。本研究首先对羧酸酯酶基因(CarE4)进行了克隆,并将其插入到pCold TF DNA Vector中,在大肠杆菌中进行了原核表达,最后用疏水层析和离子交换层析方法对目的蛋白进行了纯化。本文成功建立了羧酸酯酶蛋白原核表达和纯化技术体系,为进一步研究东亚飞蝗羧酸酯酶的生理功能、结构特点和作用原理提供了基础资料。     

Lipoprotein transformations during adipokinetic hormone action in Locusta migratoria

ABSTRACT. Detailed studies of lipoprotein A⁺ formation during AKH action have been made in Locusta migratoria L. using gel filtration combined with the use of radiolabeled haemolymph protein probes. In addition, we have assessed the quantitative contribution of the C_L-proteins to A⁺ formation by direct measurement of the changes in concentration of free C_L-proteins and report some properties of the C-I and C-II proteins: they appear to be glycoproteins of 20,000 and 16,000 MW respectively, but do not bind to concanavalin A. We have confirmed earlier observations (using different techniques) which showed that liproprotein A_yellow is not involved per se in A⁺ formation during the first 15 min of AKH action. In contrast, the (two) C_L-proteins take part in A⁺ formation without any apparent delay after hormone injection. Our observations show that A⁺ formation is essentially complete within 30 min of AKH injection, although further C_L-protein binding and lipid-loading do occur subsequently. After 30 min there is no further decrease in the A_yellow titre. It is argued that much, if not all, C_L-protein is located at the surface of the A⁺ particle. From the changes in titres which occur in A_yellow and C_L-proteins during AKH action we estimate that A⁺ is formed from 1 mole of A_yellow and approximately 28 moles of C_L-proteins. Using these figures we calculate an apparent molecular weight for A⁺ within the range of 1.65–2.12×10⁶, which is in reasonable agreement with estimates derived from gel exclusion chromatography data. These studies emphasize the dynamic and fully reversible nature of lipoprotein A⁺ formation and highlight the complex nature of the lipoprotein transformations occurring during hormone-stimulated lipid transport in locusts.