Effective control of a field population of Helicoverpa armigera by using the small RNA virus Helicoverpa armigera stunt virus (Tetraviridae: Omegatetravirus)

The first comprehensive field trial using an insect small RNA virus as a control agent on a cropping system was conducted with the Helicoverpa armigera stunt virus (family Tetraviridae, genus Omegatetravirus, HaSV). The virus was semipurified, quantified, and applied at two rates, 4 x 10(15) and 4 x 10(14) virus particles/ha, with minimal formulation on sorghum against the bollworm Helicoverpa armigera (Hübner). For comparison, a commercial preparation of Helicoverpa zea single-nucleopolyhedrovirus (HzSNPV, Gemstar) was applied at the same time at 9.27 x 10(11) polyhedral inclusion bodies/ha. The HaSV application rates were determined by a novel procedure using laboratory LC50 bioassay data for HaSV and HzSNPV and calibration to the known field application rate of the HzSNPV. The baculovirus and the higher rate of HaSV produced statistically equivalent reductions in the larval populations of around 50% at both 3 and 6 d postapplication (dpa) compared with untreated plots. The 10-fold lower rate of HaSV reduced the larval population by 50% at 3 dpa and approximately 30% at 6 dpa. Persistence of HaSV over a 72-h period was found to be similar to that of HzSNPV, although the amount of HaSV available on the sorghum heads increased at 130 h postapplication, due most likely to dispersal of newly produced virus from cadavers and frass. The results from this trial indicate that HaSV could be used as an effective biopesticide for the control of H. armigera in sorghum and the ramifications for its broader use are discussed.     

The specificity of Helicoverpa armigera stunt virus infectivity.

Helicoverpa armigera stunt virus (HaSV) is a member of the Tetraviridae family of RNA viruses whose replication and expression strategies are not well understood due to the absence of an in vitro cell culture system. We set out to find such a system for HaSV by screening an array of 13 insect and 1 mammalian cell culture lines with both virus particle infection and genomic RNA transfection. No cell line was found to be permissive for replication, although entry of genomic RNA was verified. The apparent specificity of this virus for its in vivo midgut target site was strongly corroborated by studies involving Northern blots of RNA extracted from infected insects. Only larval midgut RNA showed the presence of virus after hosts were infected per os or by injection which exposed other host cell types to the virus. The absence of replication in cell culture was due to a lack, or presence, of host factors important to replicase activity and also the likely absence of virus particle binding and entry. We thus provide both in vitro- and in vivo-based evidence demonstrating that this virus is extremely specific in the type of cells in which it will initiate an infection.     

Host recognition by a polyphagous lepidopteran (Helicoverpa armigera): primary host plants, host produced volatiles and neurosensory stimulation

Abstract An important question in the host‐finding behaviour of a polyphagous insect is whether the insect recognizes a suite or template of chemicals that are common to many plants? To answer this question, headspace volatiles of a subset of commonly used host plants (pigeon pea, tobacco, cotton and bean) and nonhost plants (lantana and oleander) of Helicoverpa armigera Hübner (Lepidoptera: Noctuidae) are screened by gas chromatography (GC) linked to a mated female H. armigera electroantennograph (EAG). In the present study, pigeon pea is postulated to be a primary host plant of the insect, for comparison of the EAG responses across the test plants. EAG responses for pigeon pea volatiles are also compared between females of different physiological status (virgin and mated females) and the sexes. Eight electrophysiologically active compounds in pigeon pea headspace are identified in relatively high concentrations using GC linked to mass spectrometry (GC‐MS). These comprised three green leaf volatiles [(2E)‐hexenal, (3Z)‐hexenylacetate and (3Z)‐hexenyl‐2‐methylbutyrate] and five monoterpenes (α‐pinene, β‐myrcene, limonene, E‐β‐ocimene and linalool). Other tested host plants have a smaller subset of these electrophysiologically active compounds and even the nonhost plants contain some of these compounds, all at relatively lower concentrations than pigeon pea. The physiological status or sex of the moths has no effect on the responses for these identified compounds. The present study demonstrates how some host plants can be primary targets for moths that are searching for hosts whereas the other host plants are incidental or secondary targets.     

Polyphagy and primary host plants: oviposition preference versus larval performance in the lepidopteran pest Helicoverpa armigera

Oviposition preference and several measures of offspring performance of Helicoverpa armigera (Hübner) were investigated on a subset of its host plants that were selected for their reputed importance in the field in Australia. They included cotton, pigeon pea, sweet corn, mungbean, bean and common sowthistle. Plants were at their flowering stage when presented to gravid female moths. Flowering pigeon pea evoked far more oviposition than did the other plant species and was the most preferred plant for neonate larval feeding. It also supported development of the most robust larvae and pupae, and these produced the most fecund moths. Common sowthistle and cotton were equally suitable to pigeon pea for larval development, but these two species received far fewer H. armigera eggs than did pigeon pea. Mungbean also received relatively few eggs, but it did support intermediate measures of larval growth and survival. Fewest eggs were laid on bean and it was also the least beneficial in terms of larval growth. Among the host plant species tested, only flowering pigeon pea supported a good relationship between oviposition preference of H. armigera and its subsequent offspring performance. Australian H. armigera moths are thus consistent with Indian H. armigera moths in their ovipositional behaviour and larval performance relative to pigeon pea. The results suggest that the host recognition and acceptance behaviour of this species is fixed across its geographical distribution and they support the theory that pigeon pea might be one of the primary host plants of this insect. These insights, together with published results on the sensory responses of the females to volatiles derived from the different host plant species tested here, help to explain why some plant species are primary targets for the ovipositing moths whereas others are only secondary targets of this polyphagous pest, which has a notoriously broad host range. Handling Editor: Joseph Dickens     

Assimilatory potential of Helicoverpa armigera reared on host (Chickpea) and nonhost (Cassia tora) diets

Adaptation to plant allelochemicals is a crucial aspect of herbivore chemical ecology. To understand an insect ecology, we studied an effect of nonhost Cassia tora seed-based diet (Ct) on growth, development, and molecular responses in Helicoverpa armigera. We employed a comparative approach to investigate the proteomic differences in gut, hemolymph, and frass of H. armigera reared on a normal (chickpea seed-based, Cp) and Ct diet. In this study, a total of 46 proteins were identified by nano-LC-MS(E). Among them, 17 proteins were up-regulated and 29 proteins were down-regulated when larvae were exposed to the Ct diet. Database searches combined with GO analysis revealed that gut proteases engrossed in digestion, proteins crucial for immunity, adaptive responses to stress, and detoxification were down-regulated in the Ct fed larvae. Proteins identified in H. armigera hemolymph were found to be involved in defense mechanisms. Moreover, proteins found in frass of the Ct fed larvae were observed to participate in energy metabolism. Biochemical and quantitative real-time PCR analysis of selected candidate proteins showed differential gene expression patterns and corroborated with the proteomic data. Our results suggest that the Ct diet could alter expression of proteins related to digestion, absorption of nutrients, adaptation, defense mechanisms, and energy metabolism in H. armigera.     

The effect of parasitization by Trichogramma australicum on Helicoverpa armigera host eggs and embryos

Histological investigations of the pathology of Helicoverpa armigera (Hübner) eggs after attack by the egg parasitoid, Trichogramma australicum (Girault), indicate that the developing embryo is immediately killed by envenomation. Soon afterward the histological staining characteristics of parasitized host embryos change and the embryonic germ band dissociates into a mass of individual rounded cells. Hosts attacked by females sterilized by gamma-irradiation showed the same pathological effects as normally parasitized hosts, indicating that host degeneration is due to female venom rather than factors derived from the parasitoid embryo or larva. Cell death also occurred in older host embryos although tissue breakdown was delayed. These findings have allowed us to determine not just that the host dies but what happens to the cells and tissues, i.e., their physical appearance, the time course of their degeneration, and that the process is retarded in older hosts. These processes can possibly be emulated in artificial diets.     

Genetics of Indoxacarb resistance in Helicoverpa armigera (Hubner)

The present investigation was done with the aim of studying the genetics of Indoxacarb resistance. Selection of Helicoverpa armigera (Hubner) with Indoxacarb was done for eight generations to develop resistance. Reciprocal crosses between resistant and susceptible populations were made to understand the population genetics of Indoxacarb resistance in H. armigera . Generation-wise selection with Indoxacarb was evaluated for resistance development in H. armigera . The LC₅₀ of Indoxacarb was 2.81 p.p.m. for the first selected generation, and it increased to 272.55 p.p.m. after eight selected generations, which is a 1238.86-fold resistance compared to the susceptible strain. The estimated realized heritability (h²) after eight generations of selection with Indoxacarb was 0.45. The number of generations required for a tenfold increase in LC₅₀ (1/R) was estimated to be 2.59. The response to Indoxacarb selection in H. armigera was 0.39, the estimated selection differential (S) was 0.87, and the phenotypic standard deviation (σp) was 0.03. Reciprocal crosses between Indoxacarb resistant and susceptible strains revealed that the inheritance of Indoxacarb resistance was autosomal: neither maternal effect nor linkage was evident. The values of D_LC (0.10 and 0.09) indicated completely recessive inheritance of Indoxacarb resistance.     

Effects of nuclear polyhedrosis virus on cannibalism in Helicoverpa armigera

在室内研究核型多角体病毒(HaSNPV)感染对棉铃虫Helicoverpa armigera(Hübner)幼虫同类相残行为的影响.结果显示:感病棉铃虫随感病程度的加重,越容易被健康棉铃虫残食,而自然死亡的感病棉铃虫、冻死的感病棉铃虫和冻死的健康棉铃虫三者被健康棉铃虫残食的百分率无显著差异.表明感病棉铃虫和病虫尸体更易于被健康棉铃虫残食,是由于棉铃虫体力减弱而失去反击能力,不是由于病毒本身的影响.以健康棉铃虫、感病棉铃虫为残食者,冻死的病虫为被残食者,相残率无显著差异,表明病毒并未改变棉铃虫残食同类的天性.残食病虫的健康棉铃虫的化蛹率和羽化率均低于正常的健康棉铃虫,残食者为相残行为付出了很高的代价.     

Bt棉花对棉铃虫幼虫选择性行为的影响

利用“H”型嗅觉仪测定结果表明 :在棉花蕾期、花铃期 ,棉铃虫 2龄幼虫对常规棉的棉叶、嫩头的选择性明显高于转基因棉花品种 ,差异达显著或极显著水平。但 3龄幼虫对两者的选择性差异并不显著 ;而在棉花蕾期 ,果枝对棉铃虫 2龄幼虫选择性行为有一定的影响。试验初步说明Bt棉对棉铃虫低龄幼虫有一定的忌避作用 ,而对高龄幼虫忌避效应不显著。     

核型多角体病毒感染对棉铃虫幼虫同类相残行为的影响

在室内研究核型多角体病毒（HaSNPV）感染对棉铃虫Helicoverpa armigera（Htibner）幼虫同类相残行为的影响。结果显示：感病棉铃虫随感病程度的加重,越容易被健康棉铃虫残食,而自然死亡的感病棉铃虫、冻死的感病棉铃虫和冻死的健康棉铃虫三者被健康棉铃虫残食的百分率无显著差异。表明感病棉铃虫和病虫尸体更易于被健康棉铃虫残食,是由于棉铃虫体力减弱而失去反击能力,不是由于病毒本身的影响。以健康棉铃虫、感病棉铃虫为残食者,冻死的病虫为被残食者,相残率无显著差异,表明病毒并未改变棉铃虫残食同类的天性。残食病虫的健康棉铃虫的化蛹率和羽化率均低于正常的健康棉铃虫,残食者为相残行为付出了很高的代价。     

Bioevaluation of Subabul (Leucaena leucocephala) proteinase inhibitors on Helicoverpa armigera

Helicoverpa armigera, a highly polyphagous pest, has a broad host spectrum, causes significant levels of yield loss in many agriculturally important crops. Serine primarily responsible for most of the proteolytic activity in the larval gut of lepidopteron insects. Neonate larvae were reared on artificial diet and chickpea seeds smeared with Subabul Trypsin Inhibitor. Larvae fed with artificial diet showed reduction in larval weight up to 21% (HSTI) and 43% (LSTI). However, larvae fed on seeds showed significant reduction in weight, 52.4% (HSTI) and 60.3% (LSTI), suggesting that the diet also plays a vital role on the effectiveness of the inhibitors on larval growth and development. HSTI and LSTI inhibited the gut proteinases from larvae fed on artificial diet significantly (41.40% and 64.36%) compared to the gut proteinases (27.80% and 38.90%) from larvae fed on chickpea seeds. Seeds smeared with 10,000 TIU resulted in complete mortality of larvae while there was no mortality observed in artificial diet. The results reveal that LSTI is a stronger inhibitor of insect gut proteinases and for larvae fed with poor nutrition in the natural ecosystems, low level expression of inhibitor would be enough to affect the growth and development. Handling editor: Chen-Zhu Wang     

Differential inhibition of Helicoverpa armigera gut proteinases by proteinase inhibitors of pigeonpea (Cajanus cajan) and its wild relatives

The seeds of 36 pigeonpea [Cajanus cajan (L) Millsp.] cultivars, resistant and susceptible to pests and pathogens and 17 of its wild relatives were analysed for inhibitors of trypsin, chymotrypsin, and insect gut proteinases to identify potential inhibitors of insect (Helicoverpa armigera) gut enzymes. Proteinase inhibitors (PIs) of pigeonpea cultivars showed total inhibition of trypsin and chymotrypsin, and moderate inhibition potential towards H. armigera proteinases (HGP). PIs of wild relatives exhibited stronger inhibition of HGP, which was up to 87% by Rhynchosia PIs. Electrophoretic detection of HGPI proteins and inhibition of HGP isoforms by few pigeonpea wild relative PIs supported our enzyme inhibitor assay results. Present results indicate that PIs exhibit wide range of genetic diversity in the wild relatives of pigeonpea whereas pigeonpea cultivars (resistant as well as susceptible to pests and pathogens) are homogeneous. The potent HGPIs identified in this study need further exploration for their use in strengthening pigeonpea defence against H. armigera.     

Digestive proteolytic and amylolytic activities and feeding responses of Helicoverpa armigera (Lepidoptera: Noctuidae) on different host plants

Digestive proteolytic and amylolytic activities and feeding responses of fifth instar larvae of Helicoverpa armigera (Hübner) on different host plants including chickpea (cultivars Arman, Hashem, Azad, and Binivich), common bean (cultivar Khomein), white kidney bean (cultivar Dehghan), red kidney bean (cultivar Goli), cowpea (cultivar Mashhad), tomato (cultivar Meshkin), and potato (cultivars Agria and Satina) were studied under laboratory conditions (25 +/- 1 degrees C, 65 +/- 5% RH and a photoperiod of 16:8 [L:D] h). Our results showed that the highest protease activity in optimal pH was on cultivar Dehghan (8.717 U/mg) and lowest one was on Meshkin (3.338 U/mg). In addition, the highest amylase activity in optimal pH was on cultivar Dehghan (0.340 mU/mg) and lowest was on Meshkin (0.088 mU/mg). The larval weight of fifth instar H. armigera showed significant difference, being heaviest on Binivich (125.290 +/- 5.050 mg) and lightest on Meshkin (22.773 +/- 0.575 mg). Furthermore, the highest and lowest values of food consumed were on Goli (362.800 +/- 27.500 mg) and Satina (51.280 +/- 4.500 mg), respectively. In addition, the lowest values of prepupal and pupal weight were on Meshkin (32.413 +/- 0.980 and 41.820 +/- 1.270 mg, respectively). The results indicated that tomato (Meshkin) was unsuitable host for feeding fifth instar larvae of H. armigera.     

Exon-primed intron-crossing (EPIC) PCR markers of Helicoverpa armigera (Lepidoptera: Noctuidae)

Applying microsatellite DNA markers in population genetic studies of the pest moth Helicoverpa armigera is subject to numerous technical problems, such as the high frequency of null alleles, occurrence of size homoplasy, presence of multiple copies of flanking sequence in the genome and the lack of PCR amplification robustness between populations. To overcome these difficulties, we developed exon-primed intron-crossing (EPIC) nuclear DNA markers for H. armigera based on ribosomal protein (Rp) and the Dopa Decarboxylase (DDC) genes and sequenced alleles showing length polymorphisms. Allele length polymorphisms were usually from random indels (insertions or deletions) within introns, although variation of short dinucleotide DNA repeat units was also detected. Mapping crosses demonstrated Mendelian inheritance patterns for these EPIC markers and the absence of both null alleles and allele 'dropouts'. Three examples of allele size homoplasies due to indels were detected in EPIC markers RpL3, RpS6 and DDC, while sequencing of multiple individuals across 11 randomly selected alleles did not detect indel size homoplasies. The robustness of the EPIC-PCR markers was demonstrated by PCR amplification in the related species, H. zea, H. assulta and H. punctigera.     

Effects of Helicoverpa armigera (Noctuidae, Lepidoptera) host stages on some developmental parameters of the uniparental endoparasitoid Meteorus pulchricornis (Braconidae, Hymenoptera)

A single choice test was performed to examine developmental strategies in the uniparental endoparasitoid Meteorus pulchricornis and its host, the cotton bollworm Helicoverpa armigera. The results support the dome-shaped model in which the fitness functions are 'dome-shaped' relative to size (and age) of host at parasitism. Older and, hence, larger host larvae were simply not better hosts for the developing parasitoids. Although parasitoid size (measured as cocoon weight and adult hind tibia length) was positively correlated with host instars at parasitism, parasitoids developing in larger hosts (L5 and L6) suffered much higher mortality than conspecifics developing in smaller hosts (L2-L4). Furthermore, egg-to-adult development time in M. pulchricornis was significantly longer in older host larvae (L4-L6) than in the younger. Performance of M. pulchricornis, as indicated by fitness-related traits, strongly suggests that the L3 host is the most suitable for survival, growth and development of the parasitoid, followed by both L2 and L4 hosts; whereas, L1, L5 and L6 are the least favourable hosts. The oviposition tendency of M. pulchricornis, represented by parasitism level, was not perfectly consistent with the performance of the offspring; L2-L4 hosts, although with the same parasitism level, had offspring parasitoids with differences in fitness-related performance. Larval development in Helicoverpa armigera was usually suspended, but occasionally advanced, in the final instar.