Bioefficacy and mode of action of rocaglamide from Aglaia elaeagnoidea (syn. A. roxburghiana) against gram pod borer,Helicoverpa armigera (Hübner)

Abstract: Rocaglamide, a highly substituted benzofuran, was isolated and identified as the main biologically active component in Aglaia elaeagnoidea (syn. A. roxburghiana) for gram pod borer Helicoverpa armigera (Hübner). Addition of rocaglamide to an artificial diet retarded the growth of neonate larvae in a dose‐dependent manner with EC₅₀ values of 0.76 p.p.m. These values compared favourably with azadirachtin (EC₅₀ = 0.23 p.p.m.). However, azadirachtin was apparently more potent than rocaglamide in inducing growth inhibition via oral administration to these first stadium larvae. The candidate compound was found to have LD₅₀ and LD₉₅ values of 0.40 and 1.02 μg per larva, respectively, in topical application against third instar larvae 96 h post‐treatment. However, these values for azadirachtin were 8.16 and 25.8 μg per larva for the same period. This shows that azadirachtin was less effective against third instar H. armigera larvae in inducing acute toxicity via topical treatment in comparison with rocaglamide. However, severe morphological larval deformities were observed in such azadirachtin‐treated larvae during the process of ecdysis. The cytotoxic nature of rocaglamide was established by evaluating dietary utilization and the results did not implicate any antifeedant effect but the toxicity‐mediated effect due to reduced efficiency of conversion of ingested food. It was obvious that feeding deterrence is not the primary mode of action but a centrally mediated effect, which could be due to the induced cytotoxicity at non‐specific cellular levels.     

Bioefficacy of Ceasalpinea bonduc (L.) Roxb. against Spodoptera litura Fab. (Lepidoptera: Noctuidae)

Bioefficacy of hexane, chloroform and ethyl acetate extracts of the seeds of Ceasalpinea bonduc (L.) Roxb. was studied against third instar larvae of Spodoptera litura at 0.25, 0.5, 1, 2.5 and 5.0% concentrations. Significant antifeedant, larvicidal and pupicidal activities and least LC₅₀ values were observed in chloroform extract. The chloroform extract was subjected to fractionation using silica gel column chromatography. Six fractions were obtained; among these, the third fraction showed high antifeedant, larvicidal and pupicidal activities at 1000 ppm concentration. Abnormalities in adults were also observed. All the activities were concentration dependent. C. bonduc could be useful in integrated pest management programme.     

棉铃虫、斜纹夜蛾无公害药剂防效筛选试验

室内和田间药效试验结果均表明,辛硫磷、病毒杀虫剂、氟虫脲、除尽和高氯·辛乳油对棉铃虫和斜纹夜蛾具有良好的防治效果,适于在瓜菜上使用,可在目前海南瓜菜生产中大量推广应用。     

竹提取物对棉铃虫幼虫及菜青虫的拒食活性

研究结果表明 ,质量浓度为 1 0g L的 1 0种供试竹提取物对棉Helicoverpaarmigera铃虫均具有较强的拒食作用 ,1 2h拒食率最高达 95 . 0 3 % ,最低为 74. 0 7% ,其中毛金竹 (Phyllostachysnigravar.henonis)、白纹短穗竹 (Brachystachyumalbostriatum)提取物对棉铃虫 3龄幼虫的AFC50 (拒食中浓度 )分别为 2 . 0g L、2. 7g L ,但 2 4h、48h拒食率均明显下降。对菜青虫 3龄pierisrapae幼虫的拒食效果以凤凰竹 (Bambusamultiplex)提取物最好 ,2 4h拒食率为 71. 69% ;对 4龄菜青虫的拒食效果以毛金竹提取物最好 ,拒食率为71 . 45 % ,其中毛金竹、白纹短穗竹 2种竹提取物对菜青虫 3龄幼虫的AFC50 分别为 2 .68g L和 3 . 3 7g L。研究结果对于开发环境友好农药以及充分利用竹类资源均具有重要意义。     

杀菌剂丙环唑对斜纹夜蛾的毒性

在研究比较了11种农药对斜纹夜蛾Spodoptera litura细胞（简称SL细胞）的毒杀活性基础上,选择毒杀活性最高的杀菌剂丙环唑,对其毒理学机理进行进一步研究。结果表明,丙环唑的细胞毒力最高,在100 μg/mL浓度下处理后48 h,SL细胞的死亡率为98.08％。处理后36 h,丙环唑对SL细胞的LC₅₀值为20.31 μg/mL。丙环唑能明显降低SL细胞的蛋白质含量。以0.5 μg/头的丙环唑注射斜纹夜蛾4龄幼虫,处理后72 h,试虫血淋巴总含量及血细胞数分别下降了26.80％和25.26％;在1.0 μg/头的剂量下,则分别下降了37.67%和36.32%。以0.5 μg/头和1.0 μg/头的丙环唑注射处理后,斜纹夜蛾幼虫体重显著降低。此外,丙环唑能降低斜纹夜蛾幼虫血淋巴含糖量及血淋巴蛋白质含量。在注射处理后96 h和120 h,丙环唑对斜纹夜蛾4龄幼虫的LD₅₀值分别为0.59 μg/头和0.45 μg/头。丙环唑对SL细胞和斜纹夜蛾幼虫均具有较好的毒杀活性,显示出丙环唑类似物控制害虫的可能性。     

Phagodeterrence and insecticidal activity of Hyptis suaveolens (Poit.) against four important lepidopteran pests

This study was carried out to evaluate the antifeedant and larvicidal activities of Hyptis suaveolens Poit. (Lamiaceae) leaves crude extracts and their fractions against four lepidopteran pests namely Helicoverpa armigera (Hbn.), Spodoptera litura (Fab.), Earias vittella (Fab.) and Leucinodes orbonalis (G.). Hexane, chloroform and ethyl acetate extracts were tested by leaf disc and fruit disc no-choice methods at 1% and 1,000?ppm concentrations for crude and fractions, respectively. Ethyl acetate extract of H. suaveolens exhibited the maximum antifeedant and insecticidal activity at 1% concentration against all the tested insects. It was subjected to fractionation using silica column chromatography with different combinations of hexane and ethyl acetate used as mobile phase. Among the 15 fractions obtained, fraction 2 showed the maximum antifeedant and insecticidal activity against all tested insects at 1000?ppm concentration. Preliminary phytochemical analysis of fraction 2 showed the presence of terpenoids and alkaloids. H. suaveolens could be considered as a safe and eco-friendly insecticide for lepidopteron pest management.     

Antifeedant and toxic activity of Trichilia americana extract against the larvae of Spodoptera litura

The biological activity of a crude methanolic extract of Trichilia americana (Sesse and Mocino) Pennington (Meliaceae) was assessed using the Asian armyworm, Spodoptera litura (Fabr.) (Lepidoptera: Noctuidae). The extract exhibited strong antifeedant activity in a choice leaf disc bioassay with 0.18 g cm^–2 extract deterring feeding by 50%. In nutritional assays, the crude extract reduced growth, consumption and the utilisation of ingested and digested food in a dose-dependent manner when fed to larvae, suggesting both antifeedant and toxic activities. When relative growth rates were plotted against relative consumption rates, the growth efficiency of the S. litura fed on diet containing T. americana crude extract was significantly less than that of control larvae. This result further indicates that the extract acts as both an antifeedant and chronic toxin. Toxicity is only seen following ingestion and was not observed following topical application or injection into the hemocoel. Larvae reared initially on extract-containing diet then transferred to control diet showed nutritional indices comparable to those of larvae fed continuously on control diet. This suggests that the extract is not permanently damaging the insect's digestive tract. The mode-of-action of the extract as a chronic toxin remains unknown.     

复配杀虫剂BtA对棉铃虫的杀虫效果及对天敌中红侧沟茧蜂蛹和成虫的触杀毒性(英文)

为了科学施药, 合理保护和利用天敌对棉铃虫Helicoverpa armigera进行综合防治, 本研究在室内测定和比较了复配杀虫剂BtA和7种常用杀虫剂（阿维菌素、 Bt、 β-氯氰菊酯、辛硫磷、定虫隆、氰戊菊酯和丁硫克百威）对棉铃虫的杀虫效果及对其天敌中红侧沟茧蜂Microplitis mediator蛹和雌成虫的触杀毒性。采用浸叶法测定杀虫剂在室内72 h内对棉铃虫3龄幼虫的杀虫效果, 结果表明： 与其他杀虫剂相比, BtA对棉铃虫幼虫具有更高的毒性和致死效应 (LC₅₀=0.7364 mg/mL)。将棉铃虫3龄幼虫接到用浓度4 mg/mL上述杀虫剂分别处理过的大白菜上24, 48和72 h, 发现其死亡率之间存在显著差异（P≤0.05）。但在72 h后, BtA和其他杀虫剂对棉铃虫幼虫的杀虫效果之间并无显著差异(P>0.05)。BtA施药后, 随着时间的延长, 棉铃虫幼虫的死亡率也在增加。另外, 通过杀虫剂对寄生蜂中红侧沟茧蜂蛹和雌成虫的触杀毒性的生物测定发现： 与β-氯氰菊酯、 氰戊菊酯和丁硫克百威相比, BtA对中红侧沟茧蜂蛹和雌成虫的毒性较低, 对其蛹和雌成虫致死率分别仅为13.82%和7.33%。本研究证明BtA对鳞翅目害虫具有中等毒性, 而对寄生蜂中红侧沟茧蜂则具有较低毒性。     

Identification, isolation and characterization of the antifeedant constituent of Clausena anisata against Helicoverpa armigera (Lepidoptera: Noctuidae)

Abstract  Hexane, petroleum ether, chloroform, ethyl acetate, methanol and water extracts of Clausena anisata [(Willd.) Hook F. Ex Benth] leaves and roots were evaluated against Helicoverpa armigera (Hübner) for antifeedant activities. Antifeedant activity was confirmed, and was found to be higher in root extracts than those of the leaf. Chloroform and petroleum ether extracts of the root showed strongest antifeedant activities (DC₅₀s [concentration (C) causing 50% deterrence compared with the control] 0.014% and 0.016% respectively), and root extracts were fractionated using silica gel column chromatography. One fraction of the chloroform and one of the petroleum ether root extracts was active; and on the basis of mass spectroscopy and ¹H and ¹³C nuclear magnetic resonance spectral data, the active compounds in the two fractions were confirmed to be identical, and identified as osthol [2H-1-Benzopyran-2-one, 7-methoxy-8-(3-methyl-2-butenyl)]. The highest concentration of osthol was found in the chloroform root extract. Antifeedant activities of the root extracts, as measured by DC₅₀ values, were highly correlated with their osthol contents. Approximately 99% of the variation in bioactivity of the root extracts could be accounted for by variation in osthol content; osthol therefore, appeared to be an antifeedant component of C . anisata to H. armigera . This may provide a new approach to managing this pest.     

番荔枝内酯化合物布拉它辛对斜纹夜蛾的杀虫活性及对SL细胞的致凋亡作用

为了明确番荔枝内酯化合物布拉它辛的杀虫活性和探索杀虫作用机理,本研究采用浸叶法测定该化合物对斜纹夜蛾Spodoptera litura幼虫的生物活性,采用MTT检测法和流式细胞术,研究该化合物对斜纹夜蛾离体培养卵巢细胞（SL细胞）的细胞毒力和致细胞凋亡作用,以及对SL细胞线粒体膜电位的影响。结果表明: 布拉它辛不仅对斜纹夜蛾幼虫具有良好的拒食活性,处理后24 h,对2龄和3龄幼虫的AFC₅₀值分别为60.25 μg/mL和86.73 μg/mL,还对幼虫生长有良好的抑制作用。布拉它辛处理SL细胞后24 h和48 h,IC50值分别为22.32 μg/mL和10.03 μg/mL。流式细胞仪检测结果表明,布拉它辛对SL细胞具有明显的致凋亡作用,可导致SL细胞线粒体膜电位显著下降。本研究表明布拉它辛对斜纹夜蛾幼虫具有良好的拒食活性与生长抑制作用, 并且该化合物能明显抑制SL细胞的增殖,诱导细胞凋亡,降低线粒体膜电位。因此,布拉他辛具有广阔的研究应用前景。     

黄皮种子提取物对斜纹夜蛾幼虫的杀虫活性及有效成分鉴定

【目的】测定黄皮Clausena lansium种子提取物对斜纹夜蛾Spodoptera litura幼虫的拒食活性,对有效成分进行分离、鉴定,并进一步测定其拒食活性、生长发育抑制活性及细胞毒性,为筛选斜纹夜蛾生态防治植物源杀虫剂提供依据。【方法】采用叶蝶法测定黄皮种子甲醇提取物及其石油醚、乙酸乙酯、正丁醇和水4种溶剂萃取物对斜纹夜蛾3龄幼虫的非选择性拒食活性和生长发育抑制活性;用硅胶柱层析进行石油醚萃取物和乙酸乙酯萃取物拒食活性成分分离;用核磁共振谱(NMR)进行拒食有效成分结构鉴定;用MTT法测定拒食活性成分对SL细胞毒性。【结果】黄皮种子甲醇提取物对斜纹夜蛾3龄幼虫具有较强的拒食活性,活性成分主要分布于石油醚相和乙酸乙酯相中;石油醚相和乙酸乙酯相经硅胶柱层析分离得10个馏分,以黄色晶体馏分2的拒食活性最强;经分离纯化及核磁共振氢谱和碳谱鉴定,黄皮新肉桂酰胺B(lansiumamide B)为主要的拒食活性成分,对斜纹夜蛾3龄幼虫48 h拒食中浓度为214.95μg/m L,同时具有较强的细胞毒性,对SL细胞48 h抑制中浓度为10.09μg/m L。【结论】黄皮种子提取物对斜纹夜蛾幼虫具有较强的拒食活性及细胞毒性,黄皮新肉桂酰胺B是主要活性成分之一,作为植物源杀虫剂开发具有较好的发展前景。     

Feeding inhibition of Helicoverpa armigera （HiJbner） by Eucalyptus camaldulensis and Tylophora indica extracts

Crude leaf extracts of Eucalyptus camaldulensis and Tylophora indica were evaluated for their antifeedancy against Helicoverpa armigera larvae in no-choice bioassays. Cabbage leaf-disc dipped in different concentrations of leaf extracts was used for feeding fifth instars of H. armigera for 24 hours. Ethanol extracts from both the plant species exhibited significant feeding inhibition, activity. The effective concentrations of ethanol extracts that caused 50% reduction in larval feeding （EI50） were 6.9% for E. camaldulensis and 2.8% for T. indica, while these values of hexane extracts were 9.3% and 5.2%, respectively. Compared to the crude leaf extracts, crude alkaloids from T. indica and crude tannins from E. camaldulensis were more potent in reducing larval feedings.     

Host crop influence on the susceptibility of the American bollworm, Helicoverpa armigera, to Bacillus thuringiensis ssp. kurstaki HD-73

Studies on the susceptibility of F₁ neonates of Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) collected from chickpea in Delhi and cotton in Punjab, Haryana and Rajasthan in northern India, to Bacillus thuringiensis ssp. kurstaki HD‐73, and the impact of host crop diets on insect susceptibility, were carried out by diet incorporation bioassays. The susceptibility of F₁ neonates of H. armigera to Bacillus thuringiensis ssp. kurstaki HD‐73 ranged from twofold (LC₅₀ 96 h, 84.5–164.2 µg (ai) l^?1) for chickpea to about fivefold (LC₅₀ 96 h, 51.1–247.7 µg (ai) l^?1) for cotton. The F₁ neonates of insects collected from pearl millet were twice as tolerant as those collected from cotton and sunflower at Sirsa to B. thuringiensis ssp. kurstaki HD‐73, suggesting that there was an influence of host crops on insect susceptibility. Insects originally collected from cotton fields at Bhatinda and reared for four generations on a chickpea‐based meridic diet were used to initiate host‐specific colonies of H. armigera. These host‐specific colonies were allowed to complete one generation on meridic diets prepared with different hosts, viz., cabbage, cauliflower, chickpea, green pea, pearl millet, and pigeon pea. Larvae of H. armigera were heaviest on the 15th day, and had a higher growth rate on a pigeon pea‐based diet than all other host diets. The larval period was shorter on chickpea and pigeon pea, with higher percentage pupation than all other host‐diets. The pupal weight of H. armigera was greater on chickpea and pigeon pea diets than on other host diets. The growth and development of larvae was significantly poorer on pearl millet diet than on other host diets. The F₁ neonates of H. armigera belonging to cabbage, cauliflower, and pearl millet host‐specific colonies were more susceptible than those belonging to chickpea, green pea, and pigeon pea host‐specific colonies to B. thuringiensis ssp. kurstaki HD‐73, suggesting the importance of proteinaceous nutrients in tolerance. The F₁ neonates of the pearl millet colony of H. armigera grown on a chickpea‐diet for 4 days were significantly more tolerant to B. thuringiensis ssp. kurstaki HD‐73 than those reared on the pearl millet‐based diet. These studies show the impact of the host diet of H. armigera on tolerance to B. thuringiensis.     

Baseline toxicity of emamectin and spinosad to Helicoverpa armigera (Lepidoptera: Noctuidae) for resistance monitoring

Acute toxicity studies of emamectin and spinosad against Helicoverpa armigera revealed that the pest is highly susceptible to both the insecticides. The median lethal dose (LD₅₀) of emamectin is 3.86 × 10⁻³ µg per larva. The median lethal concentrations (LC₅₀) of emamectin and spinosad were found to be 0.09 and 2.94 ppm, respectively. The discriminating doses were fixed based on the LC₉₅ of the susceptible population of H. armigera as 0.80 ppm for emamectin and 10 ppm for spinosad. Resistance was not observed when the discriminating doses of emamectin and spinosad were applied on field-collected populations of H. armigera from two intensive cotton growing areas, Coimbatore and Madurai, India.     

印楝素乳油对斜纹夜蛾的生物活性及田间防效研究

在室内测定了印楝素乳油对斜纹夜蛾(Spodopteralitura)的生物活性,并进行了田间防治试验.结果表明,印楝素乳油对斜纹夜蛾具有多种生物活性,主要表现在对幼虫的拒食作用、生长发育的抑制作用和一定的毒杀活性,以及对成虫产卵的忌避作用.经印楝素处理后,幼虫平均发育历期延长、幼虫体重、蛹重、化蛹率和蛹羽化率显著低于对照,印楝素乳油12μl·L^-1浓度处理3龄幼虫5和7d后的体重分别是对照组的36.8%和14.5%,而幼虫平均发育历期比对照组延长6d左右,化蛹率和蛹羽化率分别是27.79%和37.23%,显著低于对照组85.66%和82.11%.印楝素对低龄幼虫有一定的毒杀活性,而对高龄幼虫活性则很低.田间防治试验表明,印楝素乳油对斜纹夜蛾种群有良好的控制作用,施药7d后其防治效果要优于化学农药10%除尽悬浮剂.     

棉铃虫和甜菜夜蛾中肠丝氨酸蛋白酶活性测定以及活化、降解CrylAc毒素分析

昆虫中肠对Bt原毒素活化与对活化毒素降解的变化被认为是害虫对Bt产生的机制之一,研究比较棉铃虫Helicoverpa armigern（Hǔbner）与甜菜夜蛾Spodoptera exigm（Hǔbner）的中肠液、BBMV蛋白酶的活性,通过SDS-PAGE分析2种昆虫对原毒素的活化速度与对活化毒素的降解速度。2种昆虫的中肠液蛋白酶活性均显著高于BBMV蛋白酶活性,中肠液与BBMV均能迅速活化原毒素并继续降解活化后的毒素,与中肠液相比,BBMV对原毒素的活化与对活化毒素的降解均慢于中肠液,甜菜夜蛾对毒素的活化与降解又慢于棉铃虫。另外,还测定抑制剂对中肠液蛋白酶活性的抑制作用,结果表明,各抑制剂对棉铃虫和甜菜夜蛾相应酶活性的抑制表现出相同的趋势,TLCK对丝氨酶蛋白酶具较好的抑制作用,而PMSF对胰蛋白酶的抑制作用次之,TPCK对胰凝乳蛋白酶的抑制作用较弱。     

双酰胺类杀虫剂对棉铃虫和松毛虫赤眼蜂的选择毒力

为了指导棉铃虫防控中化学杀虫剂和天敌昆虫协调使用,本文采用浸叶法和玻管药膜法测定了4种双酰胺类杀虫剂对棉铃虫3龄幼虫和松毛虫赤眼蜂成蜂的毒力.结果表明,供试双酰胺类杀虫剂中氯虫苯甲酰胺、氟苯虫酰胺和氟氰虫酰胺对棉铃虫幼虫表现出了较高的毒力水平(LC50分别为0.2882、0.3894、0.8609 mg/L),稍低于对照药剂甲氨基阿维菌素苯甲酸盐(简称:甲维盐)(LC50为0.06493 mg/L);而溴氰虫酰胺对棉铃虫的毒力(LC50为3.7638 mg/L)明显低于甲维盐.从对天敌昆虫安全性来看,氯虫苯甲酰胺、氟苯虫酰胺和氟氰虫酰胺对松毛虫赤眼蜂的毒性(LC50分别为1.0060、0.9933、1.1541 mg/L)明显低于甲维盐(其LC50为0.1646 mg/L),表现出了较高的安全性;但是,溴氰虫酰胺对松毛虫赤眼蜂毒性较高(LC50为0.4952 mg/L),安全性较差.本研究将为棉铃虫防控中药剂与天敌协调使用提供依据.