多功能氧化酶系与甜菜夜蛾对氯氟氰菊酯抗药性的关系

比较了甜菜夜蛾Spodoptera exigua (Hübner)抗氯氟氰菊酯品系和敏感品系5龄幼虫中肠、脂肪体及体壁微粒体细胞色素P450的含量,结果表明,抗性和敏感品系不同组织细胞色素P450的含量均为中肠＞脂肪体＞体壁,抗性品系中肠、脂肪体及体壁细胞色素P450的含量分别是敏感品系的1.78、1.54及1.37倍。中肠微粒体甲氧试卤灵-O-脱甲基酶、乙氧试卤灵-O-脱乙基酶、乙氧香豆素-O-脱乙基酶、芳香基羟基化酶及艾氏剂环氧化酶的活性测定结果表明,抗性品系中肠5种酶的活性分别比敏感品系的酶活性提高1.33、1.73、1.40、1.51及1.30倍,说明甜菜夜蛾对氯氟氰菊酯的抗药性与微粒体多功能氧化酶活性的提高密切相关。     

绿僵菌侵染对黏虫免疫反应的影响

为揭示黏虫被病原菌侵染后的免疫应答机制,本试验分析比较了黏虫幼虫被金龟子绿僵菌Metarhizium anisopliae侵染后不同时间其血淋巴蛋白含量、免疫关键酶系(酚氧化酶PO,溶菌酶)活性、抗菌活力以及血细胞数量等免疫相关指标的变化。结果表明:受绿僵菌侵染后,黏虫6龄幼虫血淋巴蛋白含量在侵染初期(6 h和24 h)显著高于对照,侵染48、72和96 h后幼虫血淋巴蛋白质含量显著低于对照。且侵染后幼虫血淋巴PO活性也呈现先升高后降低的趋势,侵染初期(6 h和24 h)显著高于对照,侵染后期除72 h外与对照组无显著差异。侵染初期(24 h)溶菌酶活性也显著升高,随后出现下降,并于48 h显著低于对照,但在化蛹时(96 h)再次上升。绿僵菌侵染后期,黏虫的抗菌活性下降,于侵染后72 h抗菌活性显著低于对照,而侵染初期抗菌活性与对照无显著差异。另外,绿僵菌侵染对幼虫血细胞数量以及浆血胞比率也有显著的影响,表现为:侵染后6 h和24 h血细胞总数、浆血胞比率均显著高于对照,随后与对照并无显著差异。综上,绿僵菌侵染黏虫幼虫初期会显著诱导其免疫反应的启动,体现在主要免疫指标升高,而后随着免疫系统的破坏与能量的消耗,其免疫反应能力降低,生理活动受到干扰。     

赤眼蜂培养基研究 1.蓖麻蚕幼虫和蛹血淋巴氨基酸含量的分析

在赤眼蜂离体培养所用的天然物质培养基中,昆虫血淋巴是其中致关重要的成分。在国内,昆虫血淋巴大多数取自于柞蚕蛹或蓖麻蚕蛹。分析和比较蓖麻蚕幼虫和蛹血淋巴各种氨基酸的含量及其变化,对于赤眼蜂离体培养基配方的改进是有意义的。有关蓖麻蚕血淋巴氨基酸的含量分析,国内外都已有报导。但多数只是测定其中某     

亚洲玉米螟幼虫膜结合海藻糖酶基因RNAi效应

海藻糖是昆虫的血糖，为昆虫提供能量；海藻糖酶催化海藻糖分解为葡萄糖，是几丁质生物合成的原料。围食膜（peritrophic membranes，PMs）是昆虫消化道特有结构，对昆虫消化食物、保护肠道表皮细胞具有重要作用；几丁质是PMs的重要组分。海藻糖酶（trehalase，Tre）和几丁质合成酶（chitin synthase，CHS）是几丁质合成途径的第1个和最后1个酶。本研究通过饲喂亚洲玉米螟（Orstrinia furnacalis，Asian corn borer，ACB）膜结合海藻糖酶（OfMT）基因特异的干扰dsRNA，研究RNAi对ACB幼虫中肠CHSB（OfCHSB）基因表达及幼虫发育的影响。发现处理48 h后，OfMT基因和OfCHSB基因表达量分别下降了52%和53%，幼虫发育迟缓。通过对处理及对照ACB幼虫中肠石蜡切片进行苏木精-伊红染色（hematoxylin-ethanol，HE染色）和几丁质标记，发现血腔内脂肪体组织减小、中肠围食膜组织中几丁质含量减少。推测OfMT基因有可能成为ACB的生物防治的靶标基因。     

昆虫免疫的研究(二)

１　抗菌物质的合成及调控　　Ｆａｙｅ等认为,抗菌肽的合成部位在昆虫脂肪体。Ｂｏｍａｎ等从天蚕脂肪体内分离到ｍＲＮＡ,反转录后得到ｃＤＮＡ,再经表达得到抗菌肽前体。Ｋａｋｉｍａ等人用同位素掺入的方法发现,天蚕蛹被注入细菌后只有脂肪体中３Ｈ－Ｕｒｉｄｉｎｅ掺入ＲＮＡ的量同时增加,认为抗菌肽主要合成部位在脂肪体,而在其它一些组织中亦少量合成。张双全［１］等报道,在柞蚕脂肪体中没有观察到抗菌活性,相反在生殖腺中却检测到活性。摘除生殖腺后再诱导,抗菌活力有所下降,应答时间明显滞后。他认为,昆虫的一个器官可…     

蛴螬感染嗜菌异小杆线虫后酶活性和能源物质含量变化

为探究昆虫病原线虫对蛴螬的致病机理,测定了蛴螬感染嗜菌异小杆线虫沧州品系 Heterorhabditis bacteriophora Cangzhou strain后其酚氧化酶（PO）、羧酸酯酶（CarE）活性以及血淋巴蛋白、海藻糖含量的变化。结果表明,线虫侵染后蛴螬 PO 和 CarE 活性大致表现出“升高-峰值-下降”的变化过程,华北大黑鳃金龟 Holotrichia oblita Faldermann、暗黑鳃金龟 H. parallela Motschulsky 和铜绿丽金龟 Anomala corpulenta Motschulsky 3种金龟子2龄幼虫PO活性分别在24、12、36 h达到最大并显著高于对照,CarE活性则分别在48、36、48 h达到最大且显著高于对照。注射线虫悬浮液至铜绿丽金龟3龄幼虫血腔后20 h,血淋巴蛋白含量最高且显著高于对照,28 h含量最低且显著低于对照;海藻糖含量在注射后20 h达到最高但与对照差异不显著,28 h含量最低且显著低于对照。两种酶活性以及能源物质含量的变化,表明嗜菌异小杆线虫入侵对蛴螬免疫系统和代谢过程产生了一定影响,也为揭示昆虫病原线虫对蛴螬的致病机理提供了一定的试验数据。     

昆虫病原真菌抗血淋巴免疫的机理

生物防治发展的一个重要阻力来自防治效果的不稳定,而影响防治效果的一个重要因素是寄主昆虫的免疫抵抗作用。因此,开发昆虫免疫抑制剂,研究昆虫病原真菌(虫生真菌)的抗血淋巴免疫机理,将有利于提高生物防治的效果,成为人们关注的焦点。本文从真菌分泌物对血细胞、囊泡膜型质子泵(V-H+-ATPase)、信号途径Imd等的影响及细胞壁成分的变化等方面,较详细地综述了近年来国内外对虫生真菌抗血淋巴免疫机理的研究进展,指出深入研究虫生真菌抗血淋巴免疫机理,对于明确虫生真菌的作用靶标,昆虫的免疫机制有重要意义,同时对新农药特别是免疫抑制剂的创制具有重要的理论指导和实践意义。     

昆虫病原线虫对草地贪夜蛾生理生化和组织病理学的影响

为明确昆虫病原线虫侵染对草地贪夜蛾Spodoptera frugiperda生理生化和组织病理学的影响,选择草地贪夜蛾6龄幼虫分别注入樱桃异小杆线虫Heterorhabditis beicherriana和小卷蛾斯式线虫Steinernema carpocapsae,测定草地贪夜蛾幼虫体内抗氧化酶和解毒酶活性以及能源物质含量的变化,并于显微镜下观察线虫侵染后草地贪夜蛾幼虫中肠和脂肪体组织的病理变化。结果显示,供试 2 种线虫对草地贪夜蛾6龄幼虫均有致死作用,注射剂量为2~3条侵染期线虫（infective juve-niles,IJs）/μL 的樱桃异小杆线虫和小卷蛾斯式线虫 24 h 后,草地贪夜蛾幼虫的死亡率分别为16.67%和96.67%。注射2~3 IJs/μL线虫后,草地贪夜蛾幼虫体内超氧化物歧化酶、过氧化物酶、过氧化氢酶、谷胱甘肽S-转移酶和羧酸酯酶的活性基本呈先上升后下降的变化趋势,注射樱桃异小杆线虫和小卷蛾斯式线虫后各酶活性达到峰值的时间分别为注射后24 h和12 h;血浆中总蛋白含量呈波动变化,海藻糖和游离脂肪酸的含量均逐渐降低。注射线虫后草地贪夜蛾6龄幼虫的中肠和脂肪体结构均被破坏,且小卷蛾斯式线虫引起的病变更快也更严重。表明相较于樱桃异小杆线虫,小卷蛾斯式线虫对草地贪夜蛾的生防潜能更大。     

昆虫对生物农药的抗性机制及对策

本文综述了昆虫对B.t等生物杀虫剂的抗性机制及延缓昆虫抗性发展所应采取的措施。昆虫通过下列不同机制产生抗生：1）昆虫的血淋巴对B.t等生物杀虫剂的营养细胞的抑制作用。2）各种来源的蛋白酶对毒素蛋白的过度降解作用。3）昆虫中肠沉淀蛋白对毒素蛋白的沉淀作用。4）中肠上皮修复能力增强。5）中肠的吸附位点对毒素蛋白的亲和力下降。通过加强对B.t菌株的选育，合理科学的用药方式及采用不同的模式进行植物基因操作以提高杀虫蛋白的表达和活性等综合措施，减缓和降低昆虫抗性的发展。     

苏云金杆菌对昆虫生理和代谢的影响

本文简要综述了苏云金杆菌对昆虫血淋巴和肠道，神经传导及呼吸作用的生物化学影响     

Microsomal oxidase activities in relation to age and chlorcyclizine induction in American cockroach,Periplaneta americana,fat body,midgut and hindgut

Female adult American cockroaches, Periplaneta americana L., showed definite age-dependent changes in levels of activity of the microsomal mixed-function oxidases. Cytochrome P-450 levels, EPN-detoxication, and p-nitroanisole O-demethylation activities were very low in young adult insects but increased steadily reaching a natural peak at about 100 days in fat body and at about 90 days in midgut and hindgut. The activities then declined rapidly reaching levels of young insects at about 130 to 140 days of age. NADPH-neotetrazolium-reductase activity was high in young insects, declined later in adult life, and returned to a peak at about 100 days.Injections of chlorcyclizine, a known microsomal enzyme inducer, significantly increased levels of cytochrome P-450, EPN-detoxication, p-nitroanisole O-demethylation, and NADPH-NT-reductase activities in young cockroaches. The drug injections were effective, however, only before the natural activity peak was reached. Beyond this point the injections had no inductive effect indicating that the microsomal oxidases in this insect are uninducible when normal enzyme levels are falling.NADPH-NT-reductase activity in male cockroaches, while being somewhat higher than in females, showed a similar age-dependent curve with the peak occurring at about 120 days.Age-dependent carbaryl resistance in male and female insects tended to follow levels of the microsomal oxidase activities. Fifty to 60-day-old insects, however, tended to be more resistant to the insecticide than microsomal enzyme levels would indicate.RNA levels of normal female insects showed age-dependent curves similar to those of the microsomal enzyme activities, being low in young adults and reaching a peak at about 100 days. Chlorcyclizine injections had little or no effect on total microsomal RNA levels.     

Mode of entry of oxime carbamates into insects

The outcome of our earlier work suggested that the transport of contact insecticides into insects does not involve the haemolymph, but that the chemical probably reaches the internal organs by migrating in and/or over integumental tissue. This applies to various insects having quite different integumental organisation, and to various groups of insecticides differing widely in physical and chemical characteristics. The present work has shown that carbamates (1-alkylthioacetaldoxime carbamates) with high water solubility are no exception. The permeability of the body wall of the housefly for the methyl homologue (methomyl) which is the most water-soluble of the series, although relatively high, is not high enough to account for the toxicity of the compound if it followed the haemolymph route. Methomyl when injected with water instead of an organic solvent was also found to be less toxic than the equivalent amount topically applied. Lateral migration in the integument could be demonstrated with all the carbamates tested and dissimilarities in rates were apparent. Methomyl migrated the most rapidly and the cyanomethylene homologue the most slowly. Differences in mobility may be partly responsible for inequality in toxic effect, but it is thought that metabolic detoxication is of over-riding importance.     

Effects of dieldrin and sumithion on the amino acid nitrogen and proteins in the haemolymph of desert locust Schistocerca gregaria Forsk

The changes in amino acid nitrogen, proline, and proteins in the haemolymph of desert locusts at different stages of dieldrin and sumithion posioning are reported using techniques of colorimetry, polyacrylamide gel electrophoresis, Sephadex gel chromatography, and electro-gel chromatography.Both the insecticides did not alter amino acid nitrogen and proline titers during the initial stages of poisoning. However, drastic changes were noted at the later stages. In comparison to amino acid nitrogen, the depletion in proline content was of much higher magnitude. Initially the protein content of haemolymph depleted and increased significantly at the acute poisoning stage. Several changes in the electrophoregrams and protein elution profiles were noted in the haemolymph samples of treated insects. Many new proteins were detected in treated insects and their molecular weights, diffusion coefficients, and Stoke's radii are reported.These results are discussed in relation to current ideas of protein metabolism in insects. Further, a possible role of haemolymph proteins as insecticide carriers has been postulated.     

Effect of lindane (γ-hexachlorocyclohexane) on carbohydrate and lipid reserves in the American cockroach, Periplaneta americana L

The effects of lindane on carbohydrate and lipid reserves of Periplaneta americana were studied in adult male insects. Topical application of lindane resulted in depleted levels of glycogen in the fat body (90% depletion) and thoracic musculature (57% depletion) and a 55% decrease in hemolymph trehalose (anthrone-positive material) by the prostration stage of poisoning. By contrast, lindane caused a 42% elevation of fat-body acylglycerol reserves and an associated 60% decrease of hemolymph free fatty acid levels. The lindane effects on carbohydrate and lipid were expressed also in head-ligated insects, thereby indicating that the results are not attributable solely to the action of lindane on the corpus cardiacum. The results are discussed in light of the proposal that lindane, and some other insecticides, cause indiscriminate release of neuroactive factors from the neuroendocrine system and that the consequent perturbation of physiological balance may contribute to the lethal action of the insecticide.     

Seasonal concentrations of dieldrin in water, channel catfish, and catfish-food organisms, Des Moines River, Iowa--1971-73.

Concentrations of dieldrin in aquatic insects, crayfish, minnows, and small carpsuckers, and muscle tissue of channel catfish (Ictalurus punctatus) were compared with the dieldrin content of Des Moines River water in 1971-73. Monthly mean concentrations of dieldrin in river water and most aquatic organisms were highest in June and July, soon after aldrin had been applied to corn land in the watershed. Several groups of aquatic organisms also exhibited high dieldrin levels in the fall when the dieldrin content of river water was seasonally low. The influence of temperature on metabolic rate and enzyme activity and the differences in body fat content were suggested as probable causes of variations observed in the dieldrin content of aquatic organisms.     

Phloxine B affects glycogen metabolism in larval stages of Ceratitis capitata (Diptera: Tephritidae)

Xanthene dyes have been tested as photoinsecticides on different dipteran species. Phloxine B ingested by Ceratitis capitata larvae showed low toxicity under dark conditions and acute light-dependent toxicity when the insects were exposed to light during the dispersion stage before pupariation. In this study we show that sub-lethal concentrations of Phloxine B generated subtle changes in weight gain and in the accurately regulated jumping behavior of metamorphosing larvae. These changes are correlated with a strong accumulation of glycogen in fat body, intestine and muscles, as well as with an inhibition of the glycogen phosphorylase activity. These results suggest that some of the non-phototoxic effects of Phloxine B might be caused by an alteration of the glycogen catabolism, which can eventually affect the viability of the flies.     

光肩星天牛幼虫感染绿僵菌后血淋巴中游离氨基酸含量的变化

采用高效液相色谱法测定了感染绿僵菌Metarhizium anisopliae后光肩星天牛Anoplophora glabripennis幼虫血淋巴中游离氨基酸的含量。结果表明,光肩星天牛幼虫血淋巴中共检测出 17种游离氨基酸,感染绿僵菌后不同时段,游离氨基酸总量呈现先下降后上升的变化趋势。感染24 ~72 h时,幼虫血淋巴中游离氨基酸总量与对照组相比均呈小幅下降趋势,各时间点氨基酸总量互有高低,但相差不大;感染72 h后至120 h时,游离氨基酸总量明显下降,120 h时降到最低值1 042.33 mg/100 mL;之后急剧上升,144和168 h时分别为120 h时的1.44和1.51倍。赖氨酸、缬氨酸、苯丙氨酸、精氨酸、酪氨酸、苏氨酸、异亮氨酸、组氨酸、亮氨酸、丙氨酸、甘氨酸、蛋氨酸含量的变化与游离氨基酸总量的变化趋势大体一致;而脯氨酸、丝氨酸、谷氨酸、天门冬氨酸、胱氨酸的含量呈波动变化。     

Action of bioresmethrin on the corpus cardiacum of Locusta migratoria

The action of the pyrethroid insecticide bioresmethrin on the corpus cardiacum (CC) has been studied in Locusta migratoria. Bioresmethrin increased the spontaneous electrical activity recorded from both the glandular and storage lobes, and induced repetitive discharge in these lobes in response to single electrical stimulation of the nervus corpus cardiacum II (NCC II) and NCC I, respectively. Incubation of the isolated CC in low concentrations of bioresmethrin (0.1-1.0 μmol) induced the release of bioassayable hyperlipaemic hormone. Injection of bioresmethrin into locusts was found to induce an elevation in the haemolymph lipid at a stage when there was no overt symptom of poisoning. It is concluded that bioresmethrin may act directly on the CC of locusts to modulate electrical activity and induce the release of hormones. The effects may precede overt symptoms of poisoning.     

Effect of abamectin on electrophysiological activity in the ventral nerve cord of Periplaneta americana L.: Correlation with symptoms of poisoning and levels of toxicant in vivo

Twenty-four hours after injection of adult male Periplaneta americana with 36 ng [5-³H]abamectin, when symptoms of poisoning were apparent in most individuals, similar levels of radioactivity were found in tissue samples from the ventral nerve cord, metathoracic muscle, fat body and testis of randomly-selected insects. When insects injected with [5-³H]abamectin (36 ng) were separated after 24 h into three groups showing varying symptoms, the level of radioactivity in the ventral nerve cord was found to be significantly greater in partially-paralysed and paralysed groups (4- and 7-fold respectively) than in a treated but non- paralysed group. The mean levels of abamectin in the nerve cords of unaffected, ‘partially-paralysed’ and ‘paralysed’ insects were estimated to be 4, 23 and 37 nM respectively. Extracellular studies of the in-vitro action of abamectin (10 nM and 1 μM) on the spontaneous activity of a picrotoxin-sensitive, partially desheathed, ventral nerve cord preparation of adult male P. americana found the mean response time to be relatively slow (77 and 38 min respectively). In sheathed nerve cords, the mean response time to abamectin (1 μM) was 117 min. In all cases, treatment with abamectin increased the stimulus voltage required to evoke a response. Spontaneous activity was found to be progressively reduced in ventral nerve cord preparations from ‘partially-paralysed’ and ‘paralysed’ insects compared with ‘non-paralysed’ and untreated insects following dissection 24 h after injection of P. americana with unlabelled or labelled abamectin (36 ng), while the stimulus required to evoke activity in the ventral nerve cord showed the reverse trend. These differences in electrophysiological activity could be correlated directly with varying levels of abamectin in the nerve cords of ‘paralysed’ and ‘partially-paralysed’ insects.     

4-己基间苯二酚对小菜蛾酚氧化酶抑制作用研究

以4-己基间苯二酚(4HR)为效应物,研究了其对小菜蛾酚氧化酶单酚酶和二酚酶活力的影响,旨在为寻找新型害虫控制剂提供线索。结果表明:该化合物对小菜蛾酚氧化酶单酚酶和二酚酶活力均有抑制作用,其I50值分别为0.34 μmol/L和0.50 μmol/L。该化合物对单酚酶活力表达的迟滞时间有明显的延长效应,对二酚酶的抑制作用表现为典型的可逆竞争性抑制类型,其抑制常数Ki为0.17 μmol/L。通过研究金属离子和酶液对4-己基间苯二酚吸收峰的影响后推测,该化合物对小菜蛾酚氧化酶产生的抑制作用可能与直接影响该酶活性中心的铜离子有关。