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.     

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.     

Ovicidal activity of Couroupita guianensis (Aubl.) against cotton bollworm Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae)

There is a great demand for food with increasing population of the world. Many chemical fertilisers and pesticides are used to increase crop productivity. Over use of pesticides and fertilisers has affected the environment and non-target organisms severely; minor pests have become major destructive pests. Hence, eco-friendly approaches are needed. The present study reports the effect of hexane, chloroform and ethyl acetate extract of Couroupita guianensis on the eggs of Helicoverpa armigera. All the extracts showed ovicidal activity, and among them hexane extract showed 64.28% ovicidal activity with LC₅₀ value of 2.62% and regression of (R ²) 83.5%. All the original data showed normality and homogeneity. The active hexane extract was fractionated; finally, eight fractions were obtained and studied at different concentrations. Among the fractions, fraction 8 showed 75% ovicidal activity with R ² of 90%. It showed LC₅₀ value of 322.94?ppm with χ ² value at 0.5%. C. guianensis could be used in pest management programme.     

Effects of larval host plants on over-wintering preparedness and survival of the cotton bollworm, Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae)

Laboratory colonies of cotton bollworm larvae, Helicoverpa armigera, kept at 20 degrees C under a photoperiod of L:D=10:14 were fed on five host plants (cotton, corn, kidney bean, tobacco and tomato) and an artificial diet (control) to determine the effects of larval host quality on survival and pupal over-wintering preparedness. A separate experiment showed that diapausing pupae weighed more and contained greater nutrient stores than did non-diapausing pupae. Diapausing pupae reared on different host plants showed significant differences in terms of over-wintering reserve storage, and degree of cold-hardiness (extent of low-molecular-weight substances and SCPs), and survivorship. The more nutrients the host plant had, the more the pupae weighed and the higher the levels of total lipids and glycogen. Body water content was also significantly affected by larval food quality. The mean pupal super-cooling capacities varied significantly from -16.7 to -18.9 degrees C according to host plants the larvae feed on, and these significantly related to water content, pupal weight, lipid and glycogen content, and the levels of glycerol. Levels of trehalose, glycerol, and inositol, which were mainly low-molecular-weight substances, showed no significant differences among different host plants, except for trehalose. Pupal mortality varied from 39.7% on corn to 3.3% on the artificial diet, which was significantly related to pupal weight, total lipid content, trehalose levels, and super-cooling points. These results suggest that larval food quality can affect survival and influence the over-wintering preparedness of the cotton bollworm.     

Effects of larval host plants on over-wintering physiological dynamics and survival of the cotton bollworm, Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae)

Laboratory colonies of cotton bollworm larvae, Helicoverpa armigera, were kept at 20 °C under a photoperiod of L:D = 10:14 and fed on three host plants (cotton, tobacco, kidney bean) and an artificial diet (control) to determine the dynamic effects of larval host quality on over-wintering physiology and mortality. Energy reserves (glycogen and lipid), super-cooling points (SCPs), low-molecular-weight sugars, temperature, and mortality were monitored from November 2002 to April 2003. Lipid content did not change much for each group during over-wintering, but differed according to larval host plants. Larval host plants obviously influence the amount of glycogen, as does time of year: glycogen was lowest in February and increased in early spring. During winter, the mean pupal SCPs increased the most in February, then decreased, and were also affected by larval host plant, i.e. over-wintering pupae reared on kidney bean had the highest SCPs. Levels of glycerol and inositol differed significantly among host plants and months, which peaked in February. Pupal mortality also varied according to larval host plants and time: pupae reached their highest mortality in March and showed host plant differences in January. Records show that February was the coldest month during the period we observed, which corresponded closely to changes in over-wintering characteristics.     

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.     

Ultrastructure of the larval stemmata,the stemmata nerves and the optic neuropils of the larval cotton bollworm,Heliothis armigera (Hübner) (Lepidoptera : Noctuidae)

Ultrastructure of stemmata (larval eyes), stemmatal nerves, and the optic neuropils of 5th-instar larvae of cotton bollworm, Heliothis armigera (Hübner) (Lepidoptera : Noctuidae), were examined with scanning and transmission electron microscopes. Six stemmata are on each side of the head. Each stemma consists of 7 retinula cells arranged into 2 tiers. Stemmata I and II have 4 distal retinula cells and 3 proximal cells, the other 4 stemmata (III–IV) have 3 distal cells and 4 proximal cells. Stemmata I and IV have a short proximal rhabdom and the rhabdomere of each proximal cell has its microvilli projecting in only one direction. On the other hand, each stemma (in stemmata II–V) has a long proximal rhabdom and the rhabdomere of each proximal cell has microvilli pitched in several different directions relative to the horizontal plane. An axon projects proximally from each retinula cell body. The stemmatal nerve is composed of the 42 retinular axons from all of the 6 stemmata on the same side of the head. Each stemmatal nerve projects to the ipsilateral optic neuropil. Axons from each stemma are in a fasicle (within the stemmatal nerve), which consists of 7 axons, 3–4 of them are thick and terminate synaptically in the proximal neuropil; the others are thinner and terminate in the distal neuropil. Organelles, particularly lysosomes, undergo ultrastructural transformations relative to ambient light levels. The functional significance of abovementioned structures are discussed in light of current knowledge.     

Novel polymorphic microsatellite markers developed in the cotton bollworm Helicoverpa armigera （Lepidoptera：Noctuidae）

A novel set of five polymorphic di- or trinucleofide microsatellite loci suitable for population genetic study were developed from an enriched genomic library for the pest insect cotton bollworm, Helicoverpa armigera, and cross-amplifiability of these and other published loci was tested in a closely related species, the tobacco budworm, H.assulta. The expected heterozygosity at these loci ranges from 0.62 to 0.91 in the cotton bollworm. The observed allele numbers varies from 4 to 12 in the limited number of individuals tested. Although a large proportion of cloned microsatellite sequences are present in multi-copy in the cotton bollworm genome, the overwhelming majority of the finalized polymorphic diallelic loci are tri-nucleotide microsatellites - an unexpected outcome, which should facilitate subsequent genotyping analysis.     

Structure and ultrastructure of the silk glands and spinneret of Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae)

This study provides comprehensive documentation of silk production in the pest moth Helicoverpa armigera from gland secretion to extrusion of silk thread. The structure of the silk glands, accessory structures and extrusion apparatus are reported. The general schema of the paired silk glands follows that found for Lepidoptera. Morphology of the duct, silk press, muscle attachments and spigot are presented as a three-dimensional reconstruction and the cuticular crescent-shaped profile of the silk press is demonstrated in both open and closed forms with attendant muscle blocks, allowing advances in our knowledge of how the silk press functions to regulate the extrusion of silk. Growth of the spigot across instars is documented showing a distinctive developmental pattern for this extrusion device. Its shape and structure are related to use and load-bearing activity.     

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.     

Phenotypic variability in the Old World bollworm,Helicoverpa armigera (Hübner) populations in India

Helicoverpa armigera (Hübner) is one of the most prominent polyphagous species of the Heliothinae pest complex that inflicts severe damage to a wide range of crops in India. Knowledge regarding the population structure of the pest species, whether morphological or genetic, is considered as an essential tool in making effective management decisions. Thus here, we performed the phenotypic characterization of H. armigera populations collected from varied geographic locations across India. Studied populations differed significantly for several external morphometric traits studied at larval, pupal and adult stages. Significant differences were also observed with respect to the intensity of black pigmentation on larval body as well as adult eye and forewing colour patterns. Besides external phenotypic traits, the length of genital organs like aedeagus and valva in males, and bursa copulatrix and bursa seminalis in females also differed significantly amongst populations. The dendrogram based on selected traits showed clear cut differentiation of studied populations into two major groups, one including all the South Zone populations and the other having populations from North and Central Zones. Differences based on phenotyping in the present study indicate the possibility of the existence of different subspecies within the Indian populations of H. armigera.     

Distribution of Helicoverpa armigera Hübner (Lepidoptera: Noctuidae) in tomato fields and its relationship to weather factors

A survey was conducted in tomato fields at different localities to assess the distribution of tomato fruitworm Helicoverpa armigera Hübner (Lepidoptera: Noctuidae) in Punjab Province, Pakistan. The effect of weather factors on its population was also investigated. Eleven tomato-growing districts were selected and data for larval population and fruit infestation were recorded. The maximum larval population (5.2 larvae/plant) of H. armigera was recorded in Bahawalpur district with 32.6% fruit infestation. However, the minimum larval population (1.4 larvae/plant) inflicted 14.7% fruit infestation in Rawalpindi district. The temperature was positively correlated while relative humidity showed negative interaction with the larval population and fruit infestation.     

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.     

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.     

Antifeedant activity of numb and salty taste compounds against the larvae of Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae)

Helicoverpa armigera, an important polyphagous insect pest in agriculture, attacks more than 200 plant species of more than 30 families. Our previous study showed that the choice feeding percentages of H. armigera larvae to tobacco leaf discs treated with sweet, bitter, and hot taste substances were higher than the control leaf discs, while numb and salty substances could significantly inhibit their feeding. To quantitatively determine the synergistic effect of numb and salty substances, in this paper, the antifeeding activities of numb and salty substances and their mixtures blended in different doses or volume ratios were assayed on H. armigera larvae. The first bioassay was designed to elucidate the relative feeding preference of the larvae to the leaves from several common host species, each paired with tobacco leaf discs, and the result indicated that the most preferred host leaf by the larvae was tobacco leaf, followed by cotton and peanut leaves, suggesting that tobacco leaf was the most suitable matrix for the antifeeding bioassay, and the larval consumption of maize, pepper, or tomato leaves were significantly lower than that of tobacco leaf. The second bioassay was to test the choice feeding response of H. armigera larvae to tobacco leaf discs treated with Zanthoxylum bungeanum extracts obtained with different solvents, and the result showed that the antifeeding activity of the alcohol extracts was the strongest (93.38%), and the leaf consumption in the treatment and the control showed extremely significant difference (t = 4.23, t_0.01 = 3.25, P = 0.0022), followed by the dichloromethane extracts (47.64%), while the other three solvents (water, acetone, and n-hexane) could not extract the active antifeeding components from Z. bungeanum. The larval consumption of tobacco leaf discs treated with the alcohol extracts of Z. bungeanum and NaCl solution were significantly less than their corresponding controls. The mean larval consumption of the treated leaf discs decreased with ever-increasing dosage, and the consumption of tobacco leaf discs coated with different doses of alcohol extracts of Z. bungeanum or NaCl solution showed extremely significant difference (F_{alcohol extract of Z. bungeanum} = 3.88, F_0.01 = 3.58, P = 0.0064; F_{NaCl solution} = 54.29, F_0.01 = 3.58, P = 0.0000), with maximum antifeeding effects at a dosage of 30 μL per 1.5 cm ID leaf disc. We further tested the larval consumption of tobacco leaf discs treated with alcohol extracts of Z. bungeanum in saturated NaCl solution mixed in different volume ratios, and the result showed that the choice antifeeding percentages of the treatments with 15 μL or more Z. bungeanum alcohol extracts were higher than 90%, among which the mixture with 25:15 volume ratio of Z. bungeanum alcohol extracts and saturated NaCl solution exhibited the strongest antifeeding activity, and the mean consumed leaf area of tobacco leaf discs coated with this blend was only 0.10 mm². In the further test on feeding dose-response of the 25:15 mixture, the mean leaf consumption decreased linearly with ever-increasing dosage, with a regression equation y = ?3.9356x + 120.78(R² = 0.9998), and the 30 μL dose could completely inhibit H. armigera feeding.     

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.     

Biopotential of Fleuggea leucopyrus (Koen.) Willd. against Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae)

In the past five decades, agricultural pests are controlled by synthetic pesticides which caused ill effects on non-target organisms and environment; also insect pests developed resistance and minor pests became major pests. By keeping this in mind, the present study was selected. Antifeedant and larvicidal activities and growth inhibitory effects of hexane, chloroform and ethyl acetate leaf extracts of Fleuggea leucopyrus (Koen.) Willd. against Helicoverpa armigera (Hbn.) were studied. Antifeedant and larvicidal activities were evaluated at 0.5, 1, 2.5 and 5% concentrations at laboratory conditions. Hexane and chloroform extracts at 5% concentration recorded significant antifeedant activity of 82.41 and 74.45%, respectively. Hexane extract recorded the least LC₅₀ and LC₉₀ values of 1.37 and 4.80%, respectively. Hexane extract recorded maximum larvicidal activity of 85.78% with LC₅₀ and LC₉₀ values of 1.69 and 4.94%, respectively. Ethyl acetate extract also had notable amount of larvicidal activity of 81.77% at 5% concentration. Hexane extract at 5 and 2.5% and ethyl acetate extract at 5% concentrations completely prevented the adult emergence of H. armigera. This plant could be further exploited to identify the active principle(s) responsible for the activities and to develop a novel pesticidal formulation. This is the first report of F. leucopyrus is studied for its bioefficacy against H. armigera.