Selection and heritability of resistance to Bacillus thuringiensis subsp kurstaki and transgenic cotton in Helicoverpa armigera (Lepidoptera: Noctuidae)

Compared with an unselected susceptible population, a cotton bollworm, Helicoverpa armigera (Hübner), population selected for 22 generations with transgenic cotton leaves (modified Cry1A) in the laboratory developed 11.0-fold resistance to Cry1Ac (one single-protein product MVPII). Resistance to Bacillus thuringiensis Berliner subsp kurstaki (Btk) was selected for 22 generations with a 5.2-fold increase in LC50. The estimated realized heritabilities (h2) of resistance for transgenic-cotton- and Btk-selected populations were 0.1008 and 0.2341, respectively. This reflects the higher phenotypic variation in response to Cry1Ac in the transgenic-cotton-selected population. This variation may have been caused by differences in protein toxin levels expressed in different growth stages of the transgenic cotton. Because of the different slopes of the probit regression lines between Cry1Ac and Btk, the estimated realized h2 cannot be used visually to compare resistance development to Cry1Ac and Btk in H armigera. Thus, the response quotient (Q) of resistance was also estimated. The Q values of resistance for transgenic-cotton- and Btk-selected populations were 0.0763 and 0.0836, respectively. This showed that the rate of resistance development would be similar in both selection populations. This result indicates that the selection of resistance using transgenic cotton is different from that selected using the single toxin. Resistance risk to transgenic cotton and Btk in field populations was assessed assuming different pressures of selection by using the estimated h2. Assuming the h2 of resistance in a field population was half of the estimated h2, and the population received prolonged and uniform exposure to transgenic cotton or Btk causing >70% mortality in each generation, we predicted that resistance would increase 10-fold after <23 generations for Cry1Ac in transgenic cotton-selected-populations and after <21 generations for Btk in Btk-selected populations. Cross-resistance would be expected after <48 generations for Btk in transgenic-cotton-selected populations and after <21 generations for Cry1Ac in Btk-selected population. The results show that the potential to evolve resistance is similar in both transgenic-cotton- and Btk-selected populations, but that cross-resistance development to Btk is slower in transgenic-cotton-selected populations than cross-resistance development to Cry1Ac in Btk-selected populations.     

Impact of mirid (Creontiades spp.) (Hemiptera: Miridae) pest management on Helicoverpa spp. (Lepidoptera: Noctuidae) outbreaks: the case for conserving natural enemies

Creontiades spp. (Hemiptera: Miridae) are sucking pests that attack buds, flowers and young pods in mungbeans, Vigna radiata (L.), causing these structures subsequently to abort. If left uncontrolled, mirids can cause 25-50% yield loss. Traditional industry practice has involved prophylactic applications of dimethoate to control mirids at budding and again a week later. The present trial was initiated to highlight the dangers of such a practice, in particular the risk of a subsequent Helicoverpa spp. lepidopteran pest outbreak. A single application of dimethoate halved the population of important natural enemies of Helicoverpa spp., and caused an above-threshold outbreak of Helicoverpa spp. within 11 days. This shows that even a moderate (e.g. 50%) reduction in natural enemies may be sufficient to increase Helicoverpa spp. populations in mungbeans. As a result, prophylactic sprays should not be used for the control of mirids in mungbeans, and dimethoate should be applied only when mirids are above the economic threshold. Indoxacarb was also tested to establish its effect on Helicoverpa spp., mirids and natural enemies. Indoxacarb showed potential for Helicoverpa spp. control and suppression of mirids and had little impact on natural enemies.     

Potentiation between pyrethroid and organophosphate insecticides in resistant field populations of cotton bollworm Helicoverpa armigera (Lepidoptera: Noctuidae) in Pakistan

The combined action of pyrethroids plus organophosphates was assessed on putatively resistant field populations of Helicoverpa armigera from Pakistan by using a leaf-dip bioassay. Ethion showed a good potentiation with bifenthrin, lambdacyhalothrin, cyfluthrin, betacyfluthrin, fenpropathrin, esfenvalerate, fluvalenate and tralomethrin. Profenofos was potentiating with bifenthrin but additive with lambdacyhalothrin. Methyl parathion also exhibited potentiation with bifenthrin. Contrarily, quinalphos produced an antagonism with bifenthrin. Chlorpyrifos potentiated lambdacyhalothrin in one population but had an additive effect in the other. A strong potentiation of pyrethroids by ethion in some populations indicates that esteratic detoxification is a key mechanism involved in imparting resistance to pyrethroids in Pakistani H. armigera.     

Insensitive acetylcholinesterase causes resistance to organophosphates in Australian Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae)

Organophosphates are valuable insecticides used to control Helicoverpa armigera on cotton in Australia. Those most commonly used for Helicoverpa spp. control are pro-fenofos, parathion-methyl and chlorpyrifos. However, there is an emerging organophosphate-resistance threat in Australian H. armigera, which is compounded by cross-resistance between profenofos and parathion-methyl. An insensitive acetylcholinesterase has been identified as the common resistance mechanism. No resistance to chlorpyrifos has been detected and acetylcholinesterase remains fully sensitive to the chlorpyrifos oxon. © 1998 Society of Chemical Industry     

棉铃虫载脂蛋白受体基因克隆及转录分析

载脂蛋白受体(lipophorin receptor, LpR)在昆虫体内脂质转运中发挥重要作用。为了明确棉铃虫载脂蛋白受体基因在其卵巢发育及饥饿胁迫中的功能, 本研究克隆了棉铃虫 HaLpR 基因的cDNA序列(GenBank登录号KU355886.1), 全长3 117 bp, 开放阅读框为2 643 bp, 编码880个氨基酸; 其氨基酸序列包含典型的低密度脂蛋白受体结构域, 与其他鳞翅目昆虫的LpR相似度>80%。荧光定量PCR检测结果显示, HaLpR 在雌蛾的卵巢和脂肪体组织中相对转录水平最高; 在雌蛾卵子发生期均有转录, 在雌蛾羽化后3 d表达量达到最高峰; 饥饿处理会抑制 HaLpR 在雌蛾卵巢和脂肪体中的转录。研究结果有助于解析HaLpR参与棉铃虫卵巢发育过程中的脂质转运以及应对饥饿胁迫的作用。     

Influence of three diets on susceptibility of selected insecticides and activities of detoxification esterases of Helicoverpa assulta (Lepidoptera: Noctuidae)

The oriental tobacco worm, Helicoverpa assulta Guenée, is one of the most destructive pests of tobacco and peppers in China. We determined the susceptibility of H. assulta reared on an artificial diet, chili pepper and tobacco to four insecticides (fenvalerate, phoxim, methomyl, indoxacarb) under laboratory conditions associated with the activities of acetylcholinesterase (AChE), carboxylesterase (CarE) and glutathione S-transferase (GST) in its larvae. H. assulta larvae that were fed with chili pepper were more susceptible to fenvalerate, indoxacarb, and phoxim than those that were fed with tobacco and the artificial diet, but not to methomyl. The larvae that were fed with chili pepper were 3.65-, 2.49-, 1.92- and 2.44-fold more susceptible to fenvalerate, phoxim, methomyl, and indoxacarb than those fed with tobacco, respectively. The AChE activities of H. assulta larvae that were fed with chili pepper and tobacco were 2.12 and 1.07 μmol mg⁻¹ 15 min⁻¹, respectively, almost 2-fold difference. The CarE activity of H. assulta larvae that were fed with chili pepper, tobacco and the artificial diet was 4.12, 7.40 and 7.12 μmol mg⁻¹ 30 min⁻¹, respectively. Similarly, the GST activities of H. assulta larvae that were fed with chili pepper, tobacco and the artificial diet was 52.02, 79.37 and 80.02 μmol mg⁻¹ min⁻¹, respectively. H. assulta larvae that were fed with chili pepper were more resistance to the tested insecticides. The low activities of AChE and the high activities of CarE and GST lead to H. assulta become more susceptible to the tested insecticides.     

Regional reversion of insecticide resistance in Helicoverpa armigera (Lepidoptera: Noctuidae) is associated with the use of Bt cotton in northern China

A continuous programme for monitoring resistance of Helicoverpa armigera (Hubner) to commonly used insecticides was undertaken for assessing the impact of Bt cotton planting on the evolution of the pest resistance from 1994 to 2002 in China. The bioassay results showed that the resistance levels per year in field populations of H armigera to lambda-cyhalothrin, phoxim and endosulfan decreased, respectively, to 9-15-fold, 11-14-fold and 6-8-fold in 2001-2002 from 197-262-fold, 52-74-fold and 18-38-fold before the cultivation of Bt cotton in 1997. This significant increase in susceptibility to insecticides is expected to result in a reduction in insecticide application for H armigera control in Bt cotton. It is concluded that Bt cotton is playing an important role in the long-term management of H armigera by increasing the potential for natural and chemical control of the pest.     

V-ATPase H表达量下调增强棉铃虫对Cry1Ac的抗性

棉铃虫Helicoverpa armigera是世界性重要农业害虫。目前防治棉铃虫的主要手段是种植转苏云金芽胞杆菌Bacillus thuringiensis(Bt)杀虫蛋白的转基因作物。本文旨在研究棉铃虫V-ATPase H在Cry1Ac蛋白毒力和抗性中的作用。利用实时荧光定量qRT-PCR技术分析V-ATPase H在Cry1Ac抗、感品系棉铃虫幼虫中肠及敏感品系棉铃虫幼虫受Cry1Ac诱导后的表达情况;在昆虫Sf9细胞中过表达V-ATPase H对其进行细胞定位,通过细胞毒力试验验证其对Cry1Ac毒力的影响。结果发现棉铃虫V-ATPase H基因在抗性品系中低表达,并且V-ATPase H在受到Cry1Ac诱导时也低表达;在Sf9细胞内表达V-ATPase H蛋白发现其在整个细胞中都有分布,过表达该蛋白后增强了细胞对Cry1Ac蛋白的敏感性。结果表明V-ATPase H参与Cry1Ac蛋白的毒力。     

棉铃虫V-ATPase亚基B是Cry1Ac的功能受体

目前对转苏云金芽胞杆菌Bacillus thuringiensis（Bt）的研究发现,液泡型ATP酶（Vacuolar-type proton ATPase,V-ATPase）可能是Bt的一类新型受体。我们前期通过构建棉铃虫的中肠酵母文库筛选Cry1Ac的结合蛋白发现棉铃虫V-ATPase亚基B（V-ATPase B）可以与Cry1Ac结合。为明确V-ATPase B在Cry1Ac毒力和昆虫对Cry1Ac抗性机制中的作用,本研究首先采用实时荧光定量PCR技术分析了该基因在抗感Cry1Ac棉铃虫幼虫中及其受到Cry1Ac诱导时的基因表达情况;通过Ligand blot进一步地证实了其与Cry1Ac的结合特性;并通过在Sf9细胞中表达V-ATPase B的试验验证了其功能。结果表明V-ATPase B在抗性品系及受到Cry1Ac诱导时均下调表达,Ligand blot证实了V-ATPase B与Cry1Ac特异性结合;并且在昆虫细胞内过表达该基因,会增强Cry1Ac的细胞毒力。研究结果表明棉铃虫V-ATPase B是Cry1Ac的功能受体,并有可能通过降低基因表达来参与棉铃虫对Cry1Ac的抗性形成。     

棉铃虫小分子热激蛋白sHSP22.0基因的克隆及表达谱分析

棉铃虫Helicoverpa armigera是一种重要的农业害虫,本文旨在研究小分子热激蛋白在其生长发育过程、抵御高温及Cry1Ac杀虫蛋白中的功能.利用PCR结合RACE技术克隆了棉铃虫sHSP22.0(small heat shock pro-tein 22.0)基因,通过生物信息学软件分析了棉铃虫sHSP22....     

Toxicity,persistence and efficacy of indoxacarb and two other insecticides on Plutella xylostella (Lepidoptera: Plutellidae) immatures in cabbage

Toxicities of indoxacarb on eggs and 5-day-old larvae of diamondback moth, Plutella xylostella L., on cabbage and those of field-aged leaf residues on 5-day-old larvae were determined in the laboratory. The persistence and efficacies of indoxacarb and two other newer insecticides (spinosad and emamectin benzoate) to P. xylostella were tested under field conditions. Results from laboratory bioassays indicate that indoxacarb was highly toxic to P. xylostella larvae through food ingestion, with LC₅₀ and LC₉₀ values of 24.1 and 90.1 mg AI l ^{- 1}, respectively. However, indoxacarb had no significant effects on eggs and larvae through direct contact compared with water control. The toxicity of field-aged leaf residues of indoxacarb (0-, 3-, 5-, 7-, 10-, 14-, 17- and 21-day-old residues) declined slowly and gradually under the field conditions in South Texas. Almost all larvae died on day 5 after feeding on the leaves with 0 - 14-day residue, and the mortalities were as high as 94 and 78% for the 14- and 17-day-old leaf residues. With one application, indoxacarb suppressed P. xylostella larvae below the economic threshold for 14 - 21 days. Two field trials showed that indoxacarb at 0.05 - 0.07 kg AI ha ^{- 1} was effective against P. xylostella, providing marketable cabbage with three applications per season. In addition, indoxacarb was as effective as spinosad, and significantly more effective than emamectin benzoate.     

Effects of petroleum spray oils on oviposition behaviour and larval survival of Helicoverpa armigera Hubner (Lepidoptera: Noctuidae) and Ostrinia nubilalis Hubner (Lepidoptera: Pyralidae)

We determined the effects of petroleum spray oil (PSO) (Caltex Canopy®) on oviposition responses of Helicoverpa armigera Hubner and Ostrinia nubilalis Hubner adults, and larval survival of the pest moths on cotton and maize plants in the laboratory. Application of 2% (v/v) of the PSO deterred H. armigera oviposition. Increasing the rate from 2 to 5% (v/v) did not significantly reduce the number of eggs laid by H. armigera on the treated plants. In contrast, the minimum rate at which the oil could deter oviposition of O. nubilalis on maize plants was 5% (v/v). Increasing the rate from 5 to 10% (v/v) did not significantly reduce the number of eggs laid per plant. However, a reduction in the rate of the PSO from 5 to 3% (v/v) resulted in a 73.9% increase in oviposition activity on the maize plants. In wind tunnel bioassay tests, all mated H. armigera females tested could detect and settle on plants treated with water but with plants treated with PSO at various times, only 50% of tested females settled on the plants 4?–?5 days after treatment (DAT) and none on the plants 0?–?2 DAT. A solid phase micro-extraction (SPME) test to determine the effect of the PSO on volatiles released by the cotton plants showed that the quantity of volatiles released by the cotton plants treated with PSO was lower than for water treated plants. This indicates that the PSO sprays may be suppressing or masking the leaf surface volatiles of the cotton plants, thereby deterring oviposition of H. armigera. Larval survival data show that PSO sprays can cause direct mortality of first, second and third instar O. nubilalis larvae. PSOs may have the potential to be integrated into pest management programme targeting H. armigera and O. nubilalis on cotton and maize crops, respectively.     

Potentiation/antagonism of deltamethrin and cypermethrins with organophosphate insecticides in the cotton bollworm, Helicoverpa armigera (Lepidoptera: Noctuidae)

The joint action of pyrethroids deltamethrin and cypermethrins in combination with organophosphates ethion, profenofos, chlorpyrifos, quinalphos, and triazophos was studied on putatively resistant field populations of Helicoverpa armigera from Pakistan by using a leaf-dip method. Ethion produced a good potentiation with deltamethrin, cypermethrin, alphacypermethrin, and zetacypermethrin, whereas profenofos, chlorpyrifos, quinalphos, and triazophos exhibited an antagonism with deltamethrin as well as cypermethrins. Implications of using mixtures for counteracting insecticide resistance are discussed.