菜青虫感染玫烟色拟青霉后血淋巴蛋白质含量及几种保护酶活力的变化

对玫烟色拟青霉Paecilomyces fumosoroseus侵染3、4龄菜青虫Pieris rapae后,其血淋巴中蛋白质含量、体内保护酶及谷胱甘肽S-转移酶活力进行了研究。结果表明,感病的3、4龄菜青虫血淋巴中蛋白质含量明显低于同期未感染的幼虫;感病虫体内超氧化物歧化酶、过氧化氢酶和过氧化物酶的活力发生了不同程度的改变,其中对3龄菜青虫体内酶活力的影响比4龄幼虫大。此外被侵染的3、4龄菜青虫体内谷胱甘肽S-转移酶活力在感病前期显著高于同期未感染的菜青虫,而在感病后期明显低于同期未感染的菜青虫。     

甜菜夜蛾不同龄期幼虫对药剂的敏感性及其与酶活力的关系

【目的】明确甜菜夜蛾Spodoptera exigua(Hübner)不同龄期幼虫对甲氨基阿维菌素苯甲酸盐和毒死蜱的敏感性及其与羧酸酯酶、多功能氧化酶、谷胱甘肽S-转移酶和乙酰胆碱酯酶活性的相关性。【方法】采用室内生物测定方法检测甜菜夜蛾不同龄期幼虫对药剂的敏感性,并检测了不同龄期幼虫体内羧酸酯酶、多功能氧化酶、谷胱甘肽S-转移酶和乙酰胆碱酯酶的活力。【结果】在所测定的5个龄期中,随龄期增加,敏感性逐渐降低,其中1~5龄幼虫对甲氨基阿维菌素苯甲酸盐的LC50分别为0.1010、0.3561、0.7568、1.4325和8.4390 mg/L,对毒死蜱的LC50分别为27.4632、46.8495、87.5222、129.3217和1 356.6770 mg/L。单头幼虫的羧酸酯酶、多功能氧化酶、谷胱甘肽S-转移酶和乙酰胆碱酯酶活力随虫龄的增加而提高,与龄期间对药剂的敏感性呈负相关(由于外源化合物摄入量减少,5龄单头幼虫的多功能氧化酶活力略有降低)。【结论】甜菜夜蛾不同龄期幼虫对药剂的敏感性存在非常明显的差异,在田间防治中,应选择对杀虫剂敏感性较高的低龄幼虫作为最佳防治时期。     

苏云金杆菌预处理小菜蛾对有机磷和氨基甲酸酯杀虫剂的增效作用

研究了小菜蛾Plutella xylostella幼虫经苏云金杆菌Bacillus thuringiensis预处理后,对有机磷和氨基甲酸酯杀虫剂敏感性的变化以及预处理对小菜蛾幼虫体内乙酰胆碱酯酶、羧酸酯酶、谷胱甘肽S-转移酶和谷胱甘肽的含量的影响。结果表明：苏云金杆菌预处理抗性小菜蛾幼虫后,其对甲胺磷、水胺硫磷和克百威的敏感性分别为未处理组的6.74、8.83和8.50倍;处理敏感小菜蛾幼虫后则分别为未处理组的2.96、1.69和3.88倍。苏云金杆菌预处理抗性小菜蛾,未处理组乙酰胆碱酯酶的K_m和V_max值分别为预处理组的1.86和1.56倍,所使用的6种杀虫剂对乙酰胆碱酯酶的K_I值,处理组为未处理组的1.80～2.66倍,苏云金杆菌预处理抗性小菜蛾对羧酸酯酶的K_m、K_I影响不大,但能显著地抑制羧酸酯酶和谷胱甘肽S-转移酶的活性并导致谷胱甘肽含量下降(对照分别为处理的2.02、1.76和1.66倍)。苏云金杆菌预处理敏感小菜蛾,对乙酰胆碱酯酶的K_m、V_max、K_I值和羧酸酯酶的K_m、K_I值以及谷胱甘肽含量影响不大,但能显著地抑制羧酸酯酶和谷胱甘肽S-转移酶的活性(对照分别为处理的1.54和1.64倍)。     

棉铃虫抗药性的生理生化机制研究

本文报道了棉铃虫Helicoverpa armigera田间抗性种群对杀虫剂抗药性的生理生化机制。抗性种群(HJ-R)5龄幼虫羧酸酯酶、谷胱甘肽转移酶、多功能氧化酶活力均明显高于相对敏感种群(HD-S)。两种群乙酰胆碱酯酶对杀虫剂敏感性没有显著差异。HJ-R种群的腹神经索对氰戊菊酯表现了2-3倍的神经不敏感性。HJ-R种群对氨基甲酸酯类杀虫剂的抗性主要是由代谢机制引起,其中多功能氧化酶可能起主导作用;对菊酯的抗性是由多功能氧化酶、酯酶、以及神经不敏感性几个因子综合作用的结果。     

转Bt基因抗虫玉米对亚洲玉米螟幼虫几种主要酶系活性的影响

研究了表达Cry1Ab杀虫蛋白的转Bt基因抗虫玉米对亚洲玉米螟Ostrinia furnacalis (uenée) 幼虫解毒酶、保护酶和中肠蛋白酶活性的影响,测定比较了取食转Bt基因玉米后幼虫体内α-乙酸萘酯酶、乙酰胆碱酯酶、谷胱甘肽S-转移酶、过氧化氢酶、超氧化物歧化酶、中肠总蛋白酶、类胰蛋白酶和类胰凝乳蛋白酶的活力。结果表明,取食转Bt基因玉米48 h后亚洲玉米螟幼虫体内的α-乙酸萘酯酶、谷胱甘肽S转移酶活力明显低于对照;而乙酰胆碱酯酶活力显著高于对照,在取食48 h、60 h和72 h的活力分别是对照的2.00、1.50和2.50倍。保护酶系、中肠总蛋白酶、弱碱性类胰蛋白酶和类胰凝乳蛋白酶的活性在取食48 h后明显受到抑制;但强碱性类胰蛋白酶的活性显著高于对照,取食48 h、60 h和72 h的活力分别是对照的4.00、1.67和1.33倍。乙酰胆碱酯酶和强碱性类胰蛋白酶可能与亚洲玉米螟对Bt的抗性有关。     

棉蚜抗吡虫啉品系和敏感品系主要解毒酶活性比较

通过生物测定和生物化学方法比较了棉蚜 Aphis gossypii 对吡虫啉抗性（约为7倍）和敏感品系几种主要解毒酶的活性。结果表明：氧化胡椒基丁醚对两个品系均无明显增效作用。抗性品系中羧酸酯酶和谷胱甘肽S-转移酶的比活力均明显高于敏感品系,抗性品系中羧酸酯酶的K_m值也显著高于敏感品系,说明抗性品系羧酸酯酶与底物的亲和力明显高于敏感品系。上述结果证明羧酸酯酶和谷胱甘肽S-转移酶活力增强在棉蚜对吡虫啉的抗性中起重要作用。     

β-细辛醚对谷蠹成虫体内四种酶活性的影响

采用石菖蒲Acorus gramineus根茎提取物β-细辛醚对谷蠹Rhizopertha dominica成虫进行拌粮处理,测定β-细辛醚对谷蠹体内乙酰胆碱酯酶、谷胱甘肽S-转移酶、羧酸酯酶和酯酶同工酶的时间效应和剂量效应。结果表明：β-细辛醚对试虫体内4种酶的酶活性均表现出较强的时间效应。LC₅₀（94.49 mg/kg）剂量的β-细辛醚处理后,谷蠹成虫体内乙酰胆碱酯酶和谷胱甘肽S-转移酶活性随处理时间的延长整体表现为抑制作用,对羧酸酯酶和酯酶同工酶则表现诱导增加作用。低剂量（67.5 mg/kg）β-细辛醚对乙酰胆碱酯酶具有显著的诱导作用,但是随着处理剂量的升高,对乙酰胆碱酯酶的活力多数表现为抑制作用。低剂量（≤100.0 mg/kg）β-细辛醚对谷胱甘肽S 转移酶具有诱导作用,而高剂量（≥133.3 mg/kg）β-细辛醚对谷胱甘肽S-转移酶具有抑制作用。β-细辛醚对羧酸酯酶的活性多数表现为诱导作用,提高β-细辛醚的处理剂量可提高羧酸酯酶的活力。不同剂量的β-细辛醚处理对谷蠹酯酶同工酶均具有显著的诱导作用,但诱导效果与处理剂量关系并不显著。     

球形芽孢杆菌C3-41对致倦库蚊的毒效及在蚊体内的再循环

球形芽孢杆菌Csub3-41菌株(Bacillus sphaericus C3-41)对致倦库蚊(Culex puinquefa-seiatus)幼虫有很高毒效,对2龄和3-4龄幼虫的半致死剂量(LD₅₀)分别为63.1 和89.7芽孢/蚊幼虫。处理浓度越高,取食时间越长,蚊幼虫取食到的杀蚊活性物质量越多,死亡率越高。当蚊幼虫取食亚致死剂量杀蚊活性物质后,球形芽孢杆菌在感染的活幼虫体内不增殖;但当蚊幼虫取食致死剂量杀蚊活性物质后,蚊幼死亡,球形芽孢杆菌在死蚊幼虫体内增殖明显,6天内芽孢从感染初期的1.86X10²蚊幼虫增加到1.59X10⁶/蚊幼虫。芽孢在死蚊幼虫体内能正常萌发、生长、产孢和形成毒素。增殖的芽孢同样对致倦库蚊幼虫有较高毒力。     

苦豆子生物碱对小菜蛾体内部分杀虫剂代谢酶活性的影响

以苦豆子Sophora alopecuroids 7种生物碱和小菜蛾Plutella xylostella幼虫为试材,研究了该生物碱对小菜蛾体内降解杀虫剂的羧酸酯酶、磷酸酯酶、谷胱甘肽-S-转移酶活性的影响。结果表明: 野靛碱和苦豆碱对羧酸酯酶活性有显著的抑制,为可逆抑制类型的非竞争性抑制作用;野靛碱等5种生物碱对酸性磷酸酯酶有明显的抑制,野靛碱对碱性磷酸酯酶有弱抑制作用;其中野靛碱等3种生物碱对谷胱甘肽-S-转移酶有明显的抑制作用。     

脱氧鬼臼毒素对粘虫几种代谢酶系的影响

脱氧鬼臼毒素是砂地柏Sabina vulgaris Ant.中的主要杀虫活性成分之一。为探讨其作用机理,以叶碟饲喂法处理粘虫Mythimna separata Walker 4龄幼虫,测定了饲喂处理12h、24 h、36 h、48 h和72 h后试虫的羧酸酯酶（CarE）、酸性磷酸酯酶(ACP)、碱性磷酸酯酶(AKP)、谷胱甘肽S-转移酶（GSTs）和多功能氧化酶(MFO)的活性。结果表明：脱氧鬼臼毒素对粘虫羧酸酯酶的活性无明显影响;对酸性磷酸酯酶具有明显的抑制作用,且随着处理时间的延长,抑制作用增强;对碱性磷酸酯酶的影响较为复杂,表现为先抑制,后有所恢复,再被抑制的变化过程;对谷胱甘肽S-转移酶处理12 h后活性变化不明显,24 h后被抑制,36 h后逐渐激活,与对照差异极显著;对细胞色素P450酶系的O-脱甲基酶活性具有明显的抑制作用。     

Role of antioxidant defense during different stages of preadult life cycle in European corn borer (Ostrinia nubilalis, Hubn.): Diapause and metamorphosis

Antioxidant enzymes, total glutathione (GSH), and ascorbic acid (ASA) were determined in whole body homogenates of nondiapausing larvae, diapausing larvae during the diapausing period (October, December, and February), and in pupae emerged from both diapausing and nondiapausing larvae of the European corn borer (Ostrinia nubilalis, Hubn., Lepidoptera: Pyralidae). The activities of catalase, selenium nondependent glutathione peroxidase (GPx), and glutathione-S-transferase (GST), as well as the content of GSH and ASA, were found to vary throughout the larval diapause. Compared to diapausing larvae, nondiapausing larvae were higher in levels of catalase, GPx, GST, and dehydroascorbate reductase (DHAR) activity. GSH content was also increased. However, nondiapausing larvae contained less ASA than diapausing ones. Pupae had higher GPx and GST activity and an increased ASA content compared to larvae. The pupae emerged from nondiapausing larvae had higher GST, glutathione reductase (GR), and DHAR activities, but lower GPx activity and ASA content than those emerged from diapausing larvae. Correlation analysis revealed differences in the way the antioxidant level is equilibrated for a particular stage and developmental pattern. The results suggest that cellular antioxidants are involved in both the protection of cells and the regulation of redox levels during the pre-adult stages of Ostrinia nubilalis. Arch. Insect Biochem. Physiol. 55:79-89, 2004.     

Antioxidant defense in mitochondria during diapause and postdiapause development of European corn borer (Ostrinia nubilalis, Hubn.)

Antioxidant enzymes (CAT, catalase; GPx, selenium nondependent glutathione peroxidase; GST, glutathione-S-transferase; GR, glutathione reductase; DHAR, dehydroascorbate reductase) were determined in the mitochondria of diapausing and non-diapausing larvae and pupae of both diapausing and non-diapausing larvae of the European corn borer (Ostrinia nubilalis, Hubn., Lepidoptera: Pyralidae). CAT, GST, and DHAR activity in mitochondria of diapausing larvae were reduced compared to non-diapausing larvae. Pupae of diapaused-larvae possessed lower GST, but higher DHAR activities compared to pupae of non-diapaused individuals. Comparison between larvae and pupae revealed lower GPx activity in the mitochondria of pupae. CAT activity in the mitochondria of pupae was higher compared to diapausing larvae, but lower than in non-diapausing ones. Correlation and canonical discriminant analyses revealed different antioxidant enzyme compositions for a particular stage and developmental pattern. Our results show that antioxidant enzymes have a similar role in the regulation of energetics in mitochondria as that in diapause and metamorphosis.     

Antioxidant defenses preserve membrane transport activity in Chironomus riparius larvae exposed to anoxia

Changes in enzyme activities, metabolite concentrations, and membrane transport activity underlying the Chironomus riparius larvae adaptive response to anoxia were investigated. Trehalose, malate, and aspartate degradation and alanine accumulation were recorded. During anoxia exposure, there was a boost of antioxidant defenses as shown by an increase of the specific activity of the enzymes catalase, glutathione-S-transferase, glutathione peroxidase, glutathione-synthase, malic enzyme, and NADP-dependent isocitrate dehydrogenase. The ratio, glutathione reduced over glutathione oxidized, decreased. Except for alanine and catalase, the parameters return to their basal value when larvae are transferred to normoxic conditions. To test whether antioxidant defenses had protective effects on membrane functionality, L-leucine uptake into brush border membrane vesicles and membrane lipid peroxidation was measured. No difference between membranes prepared from larvae exposed to anoxia and control larvae was found. The amino acid alanine, when present inside the vesicles, trans-stimulated leucine uptake. This effect could represent a mechanism to stimulate amino acid uptake and catabolism in vivo when free alanine concentration increases during hypoxic periods.     

Glutathione S-transferase from the spruce budworm, Choristoneura fumiferana: identification, characterization, localization, cDNA cloning, and expression.

A 23-kDa protein that was present at higher levels in diapausing 2nd instar larvae than in feeding 2nd instar larvae of Choristoneura fumiferana was purified, and polyclonal antibodies were raised against this protein. The antibodies were subsequently used to screen a cDNA library that was constructed using RNA from 2nd instar larvae. Eight identical cDNA clones were isolated. The cDNA clone had a 665-bp insert and the longest open reading frame coded for a 203-amino acid protein with a predicted molecular mass of 23.37 kDa. The deduced amino acid sequence showed high similarity to glutathione S-transferases and therefore, the cDNA clone was named C. fumiferana glutathione S-transferase (CfGST). Identity of CfGST was confirmed by using affinity-purification as well as enzyme activity assay. CfGST was closer in similarity to insect GST2 members than GST1 members. The apparent Vmax of the purified CfGST towards the substrates glutathione and 1-chloro-2,4-dinitrobenezene (CDNB) were similar. However, the enzyme had a three-fold higher affinity towards CDNB than glutathione. Analyses using Northern blot, immunoblot and immunocytochemistry demonstrated that the fat body was the major tissue where the enzyme was synthesized and stored. Higher levels of CfGST protein were present in diapausing 2nd instar larvae compared to feeding 2nd and 6th instar larvae, suggesting that besides detoxification CfGST may have other roles during insect development that are not readily apparent at present. The CfGST cDNA was expressed in a recombinant baculovirus expression system and an active enzyme was produced.     

Induction of glutathione S-transferase in the castor semilooper,Achaea janata (lepidoptera,Noctuidae) following fenitrothion treatment

Glutathione S-transferase activity was determined in the lepidopteran insect species,Achaea janata, during larval, pupal and adult stages following treatment with sublethal and lethal doses of fenitrothion. Both doses of insecticide produced significant induction of enzyme activity. The rate of induction of enzyme activity was not significantly different in insects that received sublethal and lethal doses of insecticide. Enzyme activity in the different stages of insecticide-treated insects was in the order pupa > adult > larva. However, the inducing effect of the insecticide was higher in larvae, than in pupae and adult. In the absence of induction, the level of enzyme was as much as 3 times higher in midgut tissue than in carcass. In larvae treated with sodium barbitone along with fenitrothion, the knock-down effect of the insecticide was delayed. This was attributed to the increased induction of glutathione S-transferase in the larvae treated with sodium barbitone. The level of reduced glutathione, a rate-limiting factor in the induction of glutathione S-transferase, changed in a cyclic manner in insecticide-treated larvae.     

Antioxidant enzymes of the black swallowtail butterfly,Papilio polyxenes,and their response to the prooxidant allelochemical,quercetin

The black swallowtail butterfly larvae, Papilio polyxenes, are specialist feeders that have adapted to feeding on plants containing high levels of prooxidant allelochemicals. Third, fourth, and fifth instar larvae were tested for their antioxidant enzyme activities, superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), and glutathione peroxidase (GPOX), using 850-g supernatants from whole-body homogenates. The overall antioxidant enzyme profile for P. polyxenes was high compared to other insects, with activities ranging as follows: SOD, 1.1–7.5; CAT, 124–343; GR, 1.0–7.5; and GPOX, 0 units. To determine whether these antioxidant enzymes were inducible, P. poly xenes larvae were given a prooxidant challenge by dipping parsley leaves (their diet in the initial studies) in solutions of quercetin, such that the leaves became coated with this prooxidant flavonoid. Mid-fifth instar larvae fed on quercetin-coated leaves were assayed for antioxidant enzyme activities as was previously done with the larvae fed the standard diet. Food consumption and quercetin intake were monitored. SOD activity was increased almost twofold at the highest quercetin concentration tested. CAT and GR activity, on the other hand, were inhibited by increased quercetin consumption, with GR activity completely inhibited at the highest quercetin concentration after 12 h of feeding. GPOX activity, not present in control insects, was also not inducible by a quercetin challenge. These studies point out the key role that the antioxidant enzymes play in insect defenses against plant prooxidants.     

摄入异烟肼对家蚕幼虫体内谷胱甘肽氧化还原循环和谷胱甘肽S-转移酶活性的影响

为了建立家蚕Bombyx mori的药物筛选和毒性评价模型, 以剂量为2 000 mg/kg的抗结核模药异烟肼饲喂家蚕5龄第3天幼虫后检测其中肠和脂肪体的抗氧化解毒相关代谢的变化。结果表明： 雌蚕中肠组织中, 总谷胱甘肽(GSH+2GSSG)、 还原型谷胱甘肽(reduced glutathione, GSH)和氧化型谷胱甘肽(oxidized glutathione, GSSG)含量均呈现迅速上升再缓慢下降趋势; 谷胱甘肽S 转移酶(glutathione S-transferase, GST)活性升高到较大值后逐渐降低; GSH/GSSG的比值下降表明, 在72 min后中肠组织向氧化态转移。脂肪体组织中, 总谷胱甘肽、 GSH和GSSG含量变化均呈现迅速下降再迅速上升的趋势; GST活性达到最大值后逐渐降低后趋于平稳; GSH/GSSG比值升高表明, 在72 min后脂肪体组织向还原态转移。无论雌蚕还是雄蚕, 总谷胱甘肽、 GSH和GSSG含量以及GST活性均是脂肪体高于中肠。雌蚕的总谷胱甘肽含量、 GSH和GSSG含量高于雄蚕, 但雄蚕的GST活性高于雌性。结果说明, 摄入异烟肼引起了家蚕幼虫体内谷胱甘肽氧化还原状态的改变和酶活性的变化, 在这个过程中脂肪体起主要解毒代谢作用。     

Oxidative stress in newly-hatched Chorthippus brunneus--the effects of zinc treatment during diapause, depending on the female's age and its origins

The responses of glutathione, glutathione S-transferases (GSTs), and catalase (CAT) were determined in 1-day-old larvae of Chorthippus brunneus Thunberg, 1815, a grasshopper exposed to zinc during diapause, from unpolluted (Pilica) or polluted (Olkusz, Szopienice) sites. The aim of the work was to search for differences among populations of the insects as a result of various multistress pressures in their habitats. The question of zinc toxicity in the context of energy allocation was also considered. Zinc caused a decrease in glutathione concentration in the body of zinc-treated larvae. Significant differences between control and zinc-treated groups were confirmed for young females' progeny from Pilica and Olkusz as well as old females' progeny from Olkusz. GSTs activity was generally not influenced by zinc. It is possible that GSTs were not the most important target of zinc action. On the contrary, the influence of zinc on CAT activity was found. The increase in CAT activity after zinc treatment was similar for all studied populations. An increase in CAT activity after zinc exposure seems to be the most universal reaction. CAT activity in zinc-treated grasshoppers may explain the mechanism of zinc toxicity based on reactive oxygen forms generation.     

氟虫腈对小菜蛾幼虫体内解毒酶的影响

本文从解毒酶系的角度研究了小菜蛾Plutella xylostella(L.)对氟虫腈的短期生理生化响应。用氟虫腈LC50剂量处理小菜蛾3龄幼虫一定时间后,测定小菜蛾幼虫体内多功能氧化酶(MFO)、酯酶(EST)和谷胱甘肽S-转移酶(GST)活性的动态变化。研究结果表明:经氟虫腈处理后小菜蛾体内解毒酶的活性均显著高于未经处理的对照,并且随着处理时间的延长,解毒酶活性逐渐提高。这说明氟虫腈对小菜蛾敏感品系的解毒酶具有一定的诱导作用,反之,解毒酶加快了小菜蛾体内对氟虫腈的代谢,降低了小菜蛾对氟虫腈的敏感性。     

Cymothoa indica (Crustacea, Isopoda, Cymothoidae) parasitizes the cultured larvae of the Asian seabass Lates calcarifer under laboratory conditions

Parasitic disease in fishes is one of the most important factors limiting aquaculture production and its economic viability. Cymothoa indica, a cymothoid isopod, is reported here for the first time parasitizing cultured larvae of the Asian seabass Lates calcarifer in India. Fourteen-day-old L. calcarifer larvae of mean weight 8.73 +/- 0.03 mg were fed with wild zooplankton in the laboratory. On Day 14 of rearing, larvae were found parasitized by cymothoids. Infected larvae reached a mean (+/- SE) weight of 98.86 +/- 0.30 mg, while uninfected specimens weighed 117 +/- 0.43 mg at the end of the experiment (Day 21). C. indica occurred in the branchial and anterodorsal regions of infected fish, where resultant skin lesions were red, hemorrhagic, without scales and with abundant secreted mucus. The cumulative mortality over the 3 wk period was 16.54 %. These parasites are transferred to the host via the zooplankton used as food; this could easily be overcome, either by filtering wild zooplankton to remove the infectious swimming larvae of C. indica or by using cultured copepods.