Mechanisms of premating isolation between Helicoverpa armigera (Hübner) and Helicoverpa assulta (Guenée) (Lepidoptera: Noctuidae)

Helicoverpa armigera and Helicoverpa assulta are sympatric sibling species, and in the laboratory they can interbreed and produce viable offspring. To assess the contributions of temporal barriers and sexual barriers to premating isolation, we investigated both the temporal rhythms of calling behavior and pheromone titers of H. armigera and H. assulta females and the behavioral responses of males to conspecific and heterospecific calling females in a wind tunnel. Both H. armigera and H. assulta females called throughout the scotophase, and there was more calling during the second half of the scotophase than during the first half. Maximal pheromone titer and maximal calling activity in H. armigera synchronously occurred at the sixth hour into the scotophase, whereas, in H. assulta, the maximal pheromone titer occurred 2 h before the peak of calling. Pheromone blend ratios of the two species were opposite and, within each species, changes in the ratio within the scotophase and at different ages were relatively small. Males of both H. armigera and H. assulta responded strongly to their conspecific calling females in the wind tunnel and completed the whole courtship sequence. In contrast, they did not land and had no copulation attempts in response to heterospecific calling females. These results show that the two species do not have obvious temporal differences in calling behavior and pheromone production, and the specificity of sex pheromone blend emitted by females plays a key role in their premating isolation. In addition, we summarized the potential isolation mechanisms of H. armigera and H. assulta.     

Mechanisms of monocrotophos resistance in cotton bollworm,Helicoverpa armigera (Hübner)

Insensitive acetylcholinesterase was identified as a resistance mechanism by comparing biochemical analysis with a laboratory selected monocrotophos resistant cotton bollworm (RR: 200) and the susceptible strain. The cDNA encoding AChE was cloned by the method of RACE (rapid amplification of cDNA ends). The complete AChE gene deduced from the cDNA consisted of a putative signal peptide of 32 amino acid residues, a mature protein of 615 residues, 5' untranslated regions (UTR) of 315 bp and 3' UTR of 324 bp. The coding sequence had a high degree of homology to the AChE from other insect species reported in the GenBank. After comparing analysis of the entire AChE gene sequence from 5 resistant and 6 susceptible cotton bollworm individuals, nine mutations were identified. One of them, the Ala/Thr mutation, is likely to be responsible for the AChE insensitivity to monocrotophos.     

Genetic basis of sex pheromone blend difference between Helicoverpa armigera (Hübner) and Helicoverpa assulta (Guenée) (Lepidoptera: Noctuidae)

The two closely related moth species, Helicoverpa armigera and H. assulta, are sympatric in China. Both species use a mixture of (Z)-11-hexadecenal (Z11-16:Ald) and (Z)-9-hexadecenal (Z9-16:Ald) as their sex pheromones but in widely different ratios. Hybridization and backcrossing experiments between H. armigera and H. assulta were conducted and sex pheromone compositions of the parent species, their F(1) hybrids and backcrosses were compared to study the genetic basis of the production of their sex pheromone blend composition. Results show that the difference in sex pheromone blend ratios of these Helicoverpa species is mainly controlled by an autosomal locus with two alleles, with the allele from H. armigera being almost completely dominant over that derived from H. assulta.     

Modeling Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) damages on cotton

We studied and modeled damage caused by Helicoverpa armigera larvae on cotton with the aim of developing a coupled crop pest model. Two damage components were studied: the voracity (quantity of fresh matter and number of organs consumed) and feeding preferences (type of organ infested). The laboratory no-choice study of voracity on excised squares and bolls revealed that an H. armigera larva consumes 2,856 mg of fresh matter throughout its larval life, with the sixth instar consuming 86% of this quantity. This consumption rate corresponded to 23.6 squares, or 7.8 bolls. We developed equations to predict the quantity of fresh matter uptake from an individual plant organ, according to the organ mass and the larval instar. The field study of feeding preference confirmed previous findings that larvae prefer squares to bolls, with this preference decreasing as the larval age increases. However, no significant relationship was noted between the age of larvae and the size of infested organs within each organ class (square or boll). We developed a logistic model to predict the probability of a larva infesting a boll rather than a square. According to this model, the relative organ availability in the field and the larval instar were found to be significant factors.     

Diet-delivered RNAi in Helicoverpa armigera – Progresses and challenges

Helicoverpa armigera (the cotton bollworm) is a significant agricultural pest endemic to Afro-Eurasia and Oceania. Gene suppression via RNA interference (RNAi) presents a potential avenue for management of the pest, which is highly resistant to traditional insecticide sprays. This article reviews current understanding on the fate of ingested double-stranded RNA in H. armigera. Existing in vivo studies on diet-delivered RNAi and their effects are summarized and followed by a discussion on the factors and hurdles affecting the efficacy of diet-delivered RNAi in H. armigera.     

烟青虫(Helicoverpa assulta Guenée)信息素的结构鉴定及田间试验(英语)

北京地区的烟青虫(Helicover Pa asslta Cucn(?)e)的性信息素腺体提取物经毛细管柱气相色谱分析及GC MS分析,鉴定了6种组分.这6种组分为:十六醛(16:Ald)、顺 9—十六烯醛(Z9—16:Ald)、顺11—十六烯醛(Zll 16:Ald)、顺9—十六烯醇(Z9-16:OH)、顺11—十六烯醇(Zll-16:OH)、顺9—十六烯基乙酸酯(Z9 16:OAc),比例为10.9:58.7:3.9:14.7:1.1:10.7.田间试验表明,只有16:Ald、Z9 16:Ald和Zll 16:Ald(比例为15.3:79.2:5.5)组成的三组分诱芯和Z9—16:Ald和Zll—16:Ald(比例为93.4:6.6)组成的两组分诱芯对于雄蛾有强烈的引诱活性.在3种醛为组分的诱芯中加入Z9 16:OH明显地降低引诱活性.     

Morphometric Variation in Geographic Populations of Cotton Bollworm,Helicoverpa armigera (Hübner) Occurring in South Indian Cotton Ecosystems

Morphometric characterization of twelve geographic populations of cotton bollworm, Helicoverpa armigera occurring in south Indian cotton ecosystems was done at larval, pupal and adult stages over three cropping seasons. Traits such as length and weight of larvae, pupa and length and width of the wing, length of fore-, mid- and hind femur, male reproductive organ-length of genital capsule, valves, and ejaculatory duct, female reproductive organ-length of appendix bursae and ductus bursae at adult stage were measured across three years. Populations significantly differed for most of the traits studied. It was evident that populations from northern parts recorded higher phenotypic attributes compared to those from southern parts of south Indian cotton ecosystem. Besides larval, pupal and adult external phenotypic traits, attributes of male reproductive organ viz., length of genital capsule, valves, and ejaculatory duct and female reproductive organ viz., length of appendix bursae and ductus bursae differed significantly among populations. Information on population structure and differences based on morphometry would be useful in better understanding of population dynamics and management of this pest in cotton and other crops in south India.     

Oxidases responsible for resistance to pyrethroids sensitize Helicoverpa armigera (Hübner) to triazophos in West Africa

Helicoverpa armigera (Hübner) is the major insect pest of cotton in Africa, Turkey, Asia, India, Indonesia and Australia. Populations recently developed resistance to pyrethroids in West Africa via the overproduction of cytochrome P450 (oxidases) increasing pyrethroid metabolism. One way to overcome pyrethroid resistance is to use compounds that show negative cross-resistance to pyrethroids. Triazophos is one of these compounds: it is slightly more toxic against pyrethroid resistant larvae of H. armigera than against susceptible ones. Overproduced oxidases transform the non active triazophos into its active form, triazophos-oxon, since this form was significantly more often found in larvae from pyrethroid resistant strain (23%) than in susceptible strain (15%). This suggests that oxidases, which provide resistance by degradation of pyrethroids in the resistant individuals, also activate triazophos in its toxic oxon form resulting in a negative cross-resistance.     

莱氏野村菌(Nomuraea rileyi)对棉铃虫(Helicoverpa armigera)幼虫的致病机理研究

本文利用荧光显微镜、光学显微镜及组织切片技术进行了莱氏野村菌对棉铃虫幼虫的致病过程及机理的研究。两个莱氏野村菌菌株ＥＦＤ44及ＥＦＤ38不同浓度及配方的分生孢子 ,在棉铃虫幼虫体皮上 2ｈ后开始发芽。两个菌株的分生孢子发芽方式不尽相同。真菌在侵入昆虫表皮体内前 ,菌株ＥＦＤ44及ＥＦＤ38的芽管发生了不同程度的分化。ＥＦＤ38芽管有几种分化形式 ,包括形成驸胞器结构、主芽管从孢子多处萌发、主芽管有或多或少的分枝、主芽管或分枝芽管形成楔型状等。处理后约 6ｈ ,真菌菌丝或芽管开始侵入昆虫外体皮 ,但真正成功地侵入外体皮并…     

Molecular cloning and bacterial expression of pheromone binding protein in the antennae of Helicoverpa armigera (Hübner)

A cDNA clone coding for pheromone binding protein was isolated from the antennae of Helicoverpa armigera by RT-PCR and (5'/3')-RACE technique. The full-length of H. armigera pheromone binding protein (HarmPBP) was 952 bp, possessing 162 amino acid residues including a signal peptide of 20 amino acids. Its predicted molecular weight and isoelectric point were 18.26 kDa and 5.23, respectively. This deduced amino acid sequence shared some common structural features with odorant-binding proteins from several moth species, including the six conserved cysteine motif, a typical characteristic of insect's odorant-binding proteins. Northern blot showed that HarmPBP is specifically expressed in the antennae of Helicoverpa armigera and more abundantly expressed in male than female. During the antennal development, HarmPBP is first expressed about 4 days prior to adult eclosion and rises to a plateau 2 days prior to adult eclosion. In order to obtain sufficient PBP for further determining its biochemical and physiological properties, a bacterical expression vector of PBP was constructed and successfully expressed in Escherichia coli. The recombinant PBP was shown to cross-react with an anti-PBP antiserum from Antheraea polyphemus. Polyclonal antibodies against HarmPBP were used to mark the distribution of the protein in olfactory sensilla. Very strong labeling was observed in the sensillum lymph of the hair lumen and of the sensillum-lymph cavity. In the male, HarmPBP is expressed in sensilla trichodea and not in sensilla basiconica, while in the female, it is expressed both in sensilla basiconica and sensilla trichodea.     

Population Genetic Structure of the Cotton Bollworm Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) in India as Inferred from EPIC-PCR DNA Markers

Helicoverpa armigera is an important pest of cotton and other agricultural crops in the Old World. Its wide host range, high mobility and fecundity, and the ability to adapt and develop resistance against all common groups of insecticides used for its management have exacerbated its pest status. An understanding of the population genetic structure in H. armigera under Indian agricultural conditions will help ascertain gene flow patterns across different agricultural zones. This study inferred the population genetic structure of Indian H. armigera using five Exon-Primed Intron-Crossing (EPIC)-PCR markers. Nested alternative EPIC markers detected moderate null allele frequencies (4.3% to 9.4%) in loci used to infer population genetic structure but the apparently genome-wide heterozygote deficit suggests in-breeding or a Wahlund effect rather than a null allele effect. Population genetic analysis of the 26 populations suggested significant genetic differentiation within India but especially in cotton-feeding populations in the 2006–07 cropping season. In contrast, overall pair-wise F _ST estimates from populations feeding on food crops indicated no significant population substructure irrespective of cropping seasons. A Baysian cluster analysis was used to assign the genetic make-up of individuals to likely membership of population clusters. Some evidence was found for four major clusters with individuals in two populations from cotton in one year (from two populations in northern India) showing especially high homogeneity. Taken as a whole, this study found evidence of population substructure at host crop, temporal and spatial levels in Indian H. armigera, without, however, a clear biological rationale for these structures being evident.     

不同光波及光强度下棉铃虫(Helicoverpa armigera)成虫的行为反应

运用室内行为实验方法研究了不同波长单色光和白光刺激的棉铃虫[Helicoverpa armigera (Hübner)]成虫趋光、避光反应及其与光强度之间的相互关系.结果显示:(1)一定时间暗适应的棉铃虫蛾对单色光和白光刺激均具有趋光行为反应,依其趋光反应率大小,其光谱反应曲线在340-605nm波谱范围内,表现为大小较复杂的几个峰,其中两个峰分别在蓝光区483nm、紫外340nm,另在近紫外的400nm、绿黄光区的538nm有峰.(2)一定光强度范围内,棉铃虫蛾的趋光反应率随单色光和白光光强度的增强而增大,至一定光强度时增加变缓,呈现近似S型曲线式样.(3)性别、日龄对棉铃虫蛾的光谱、光强度反应曲线均有一定影响,雌性较雄性的趋光反应率和可适应的光强度高,1、3、5日龄蛾中以3日龄的趋光反应率和可适应的光强度高.(4)棉铃虫蛾的趋光、避光反应行为具有相似的波长及光强度选择机制.     

不同光波及光强度下棉铃虫(Helicoverpa armigera)成虫的行为反应

运用室内行为实验方法研究了不同波长单色光和白光刺激刺激的棉铃虫「Ｈｅｌｉｃｏｖｅｒｐａ ａｒｍｉｇｅｒａ（Ｈｕｂｎｅｒ）」成虫趋光、避光反应及其与光强度之间的相互关系,结果显示：（１）一定时间暗适应的棉铃虫蛾对单光和白光刺激均具有趋光行为反应,依其趋光反应率大小,其光谱反应曲线在３４０－６０５ｎｍ波谱范围内,表现为大小较复杂上峰,其中两个峰分别在蓝光区４８３ｎｍ、紫外３４０ｎｍ,另外近紫外的４０ｎ     

Organophosphorus insecticides synergize pyrethroids in the resistant strain of cotton bollworm, Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) from West Africa

Helicoverpa armigera (Hübner) populations from West Africa recently developed resistance to pyrethroid insecticides through enhanced metabolism by mixed-function oxidases. The combination index method was used to study the synergism of pyrethroids by organophosphorus insecticides. Several mixtures of insecticides currently registered to control cotton pest complex in West Africa were tested, including: cypermethrin/ethion, cypermethrin/profenofos, deltamethrin/ triazophos, deltamethrin/chlorpyriphos, cyfluthrin/chlorpyriphos, and betacyfluthrin/chlorpyriphos. In the resistant strain, the organophosphorus insecticides significantly increased the toxicity of pyrethroids suppressing the resistance effect, either by additive or synergistic effects. Significant synergism was shown for the following mixtures: cypermethrin/ethion, deltamethrin/triazophos, and deltamethrin/chlorpyriphos. The use of synergism from these insecticide mixtures should prove to be an additional tool in the overall resistance management strategy because the pyrethroid resistance in H. armigera from West Africa is not yet stable, decreasing between cotton seasons and increasing with treatments. In absence of selection, the susceptibility of H. armigera to insecticides should be restored.     

In vivo and in vitro effect of Acacia nilotica seed proteinase inhibitors on Helicoverpa armigera (Hübner) larvae

Acacia nilotica proteinase inhibitor (AnPI) was isolated by ammonium sulphate precipitation followed by chromatography on DEAE-Sephadex A-25 and resulted in a purification of 10.68-fold with a 19.5 percentage yield. Electrophoretic analysis of purified AnPI protein resolved into a single band with molecular weight of approximately 18.6+1.00 kDa. AnPI had high stability at different pH values (2.0 to 10.0) except at pH 5.0 and are thermolabile beyond 80 degree C for 10 min. AnPI exhibited effective against total proteolytic activity and trypsin-like activity, but did not show any inhibitory effect on chymotrypsin activity of midgut of Helicoverpa armigera. The inhibition kinetics studies against H. armigera gut trypsin are of non-competitive type. AnPI had low affinity for H. armigera gut trypsin when compared to SBTI. The partially purified and purified PI proteins-incorporated test diets showed significant reduction in mean larval and pupal weight of H. armigera. The results provide important clues in designing strategies by using the proteinase inhibitors (PIs) from the A. nilotica that can be expressed in genetically engineered plants to confer resistance to H. armigera.     

Electrophysiological responses of female Helicoverpa armigera (Hübner) (Lepidoptera; Noctuidae) to synthetic host odours

Studies were conducted to investigate the electroantennographic (EAG) responses of adult female Helicoverpa armigera to a range of known and putative kairomone components. The studies show that at a given dose the EAG responses elicited by a series of straight-chain aliphatic primary alcohols were not dependent on volatility since butan-1-ol and pentan-1-ol elicited EAG responses that were significantly smaller than those elicited by hexan-1-ol. The amplitudes of responses to hexan-1-ol were found to be dose dependent with a dose of 10(-1) μmol at source in a non-volatile solvent eliciting the largest response. Similarly, changes in functionality in a range of C(6) straight-chain aliphatic compounds significantly changed the amplitude of response elicited, with aldehydes eliciting smaller responses than the related primary alcohols and saturated compounds eliciting higher responses than related unsaturated compounds. Of the range of nine host plant-produced terpenoids tested, ocimene and beta-phellandrene elicited the highest responses and of the six aromatic compounds tested phenylacetaldehyde and benzaldehyde elicited the largest responses, at the doses tested. The significance of these findings for analysis of floral odours by gas chromatography linked to electroantennography as a means of identifying kairomone components attractive to H. armigera are discussed.     

Characterization of three pheromone-binding proteins (PBPs) of Helicoverpa armigera (Hübner) and their binding properties

Three pheromone-binding proteins of Helicoverpa armigera were cloned and expressed in Escherichia coli. In order to characterize their physiological properties, ligand-binding experiments were performed using five biologically relevant substances including sex pheromones and interspecific signals. The results showed that one of the pheromone-binding proteins, HarmPBP1, binds strongly to each of the two principal pheromone components of H. armigera, (Z)-11-tetradecenal and (Z)-9-hexadecenal, but not to the interspecific signal (Z)-9-tetracecenal. The two remaining pheromone-binding proteins, HarmPBP2 and HarmPBP3, showed only weak affinities with the ligands tested. The 3-D structure of HarmPBP1 was predicted and the docking experiments indicate that the key binding site of (Z)-9-hexadecenal to HarmPBP1 includes Thr112, Lys111, and Phe119 whereas that of (Z)-11-tetradecenal includes Ser9, Trp37, Phe36, and Phe119.     

Karyomorphological effects of two new oil formulations on Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae)

The cotton bollworm Helicoverpa armigera is a serious pest of many economically important crops. Since this pest has become resistant to the conventional synthetic insecticides, newer compounds and formulations are being developed against this insect pest. Many natural compounds isolated from the plants were tested against this pest. Among them Hyptis suaveolens and Melochia corchorifolia showed insecticidal properties against H. arnigera. Based on bioefficacy studies, caryophyllene and β-sitosterol were isolated from H. suaveolens and M. corchorifolia respectively. The isolated natural compounds were further developed as formulations in various combinations with neem (Azadirachta indica) and karanj (Pongamia pinnata) oils. The present study indicated that the formulations influenced the karyomorphology of H. armigera.     

Bio-efficacy of different oil carriers in ultra low volume sprayer for management of pod borer,Helicoverpa armigera (Hübner) in pigeonpea ecosystem

A study was conducted to evaluate the bioefficacy of different oil carriers (sunflower oil, seamum oil, neem oil, mineral oil), sugar solution and water formulation to be used in ultra low volume sprayer (ULV) along with rynaxypyr 18.5 SC (Coragen) for the management of pod borer at the Agricultural Research Station, Gulbarga, Karnataka, India during the kharif season of 2011–2012. The results revealed that the sesamum oil carrier fits well with rynaxypyr 18.5 SC and recorded the lowest mean pod borer larval population after last spray (1.0 per five plants) the lowest pod damage (23.20%), grain damage (18.30%) and higher grain yield (9.43?q/ha) with a high B:C ratio of 3.64 in the plot sprayed with the sesamum oil carrier. The findings suggested that the sesamum oil carrier in ULV sprayer is suitable and practicable in pigeonpea ecosystem for effective management of pod borer with no risk of phytotoxicity, spray drift and operational difficulties. The foregoing studies indicated that the performance of sesamum oil carriers was superior to other carriers in terms of suppression of larval population, reduction in pod damage and harnessing higher yield.     

Changes of inheritance mode and fitness in Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) along with its resistance evolution to Cry1Ac toxin

The changes of inheritance mode and fitness of resistance in Helicoverpa armigera (Hübner) along with its resistance evolution to Cry1Ac toxin were evaluated in the laboratory. The resistance levels reached 170.0-, 209.6- and 2893.3-fold, on selection of the field population in the 16th (BtR-F(16)), 34th (BtR-F(34)) and 87th (BtR-F(87)) generation with artificial diet containing Cry1Ac toxin, respectively. As the resistance levels increased, more larvae feeding on the Bt cotton expressing Cry1Ac toxin survived. Most larvae of BtR-F(87) could develop to the 5th instar and about 3% individuals reached the adult stage. The inheritance of Cry1Ac resistance trait at three resistant levels was autosomal and incompletely recessive, but the degree of dominance decreased as the resistance increased. The resistance was primarily monogenic in BtR-F(16) strain, but polygenic as resistance increased. The relative fitness of H. armigera, measured as a ratio of R(0) (the net replacement rate) of resistant strain divided by R(0) of the susceptible strain, decreased with an increase of the resistance levels, with ratios of 0.79, 0.64 and 0.59 in their respective BtR-F(16), BtR-F(34) and BtR-F(87) strains.