Post-eclosion diuresis in adult Heliothis zea

ABSTRACT. The marked weight loss that follows adult eclosion in Heliothis zea (Boddie) (Lepidoptera: Noctuidae) was found to be the result of diuresis. This diuretic weight loss amounted to 18.2% of the emergence weight in females and 22.5% in males. Ligation between head and thorax immediately following emergence resulted in a significant reduction in this weight loss. Injection of homogenates of brain (BR) or suboesophageal ganglion (SOG) partially restored the weight loss in ligated insects, indicating potential neurohormonal regulation of this phenomenon.
Because injection of homogenates of nervous and neuroendocrine tissues other than BR and SOG did not result in increased weight loss over controls, the activity present in BR and SOG was probably a tissue-specific phenomenon. We speculate that a factor at least partially responsible for the enhanced diuretic weight loss is present in the fused BR/SOG.     

FMRF-amide immunoreactivity in a moth larva (Heliothis zea): The cerebral nervous system

The cerebral nervous system of the larval corn earworm, Heliothis zea, was examined using light microscopy and immunocytochemistry. Immunoreactivity for the molluscan cardioactive peptide FMRF-amide (FMRFa) is widely distributed. Staining is localized in perikarya and axons of the brain, subesophageal ganglion and first thoracic ganglion. Immunoreactive axons are also found in the frontal ganglion and the corpora cardiaca. Crossreactivity studies for FMRFa and bovine pancreatic polypeptide (BPP) demonstrate that (a) the immunoreactive material is more FMRFa-like than BPP-like and (b) the antigenic material is heterogeneous. For animals on different feeding regimens, immunocytochemical surveys and radioimmunoassay suggest the observed variation in immunoreactivity among individuals is unrelated to feeding state.     

Mode of transmission of nuclear-polyhedrosis virus to progeny of adult Heliothis zea

Late second-instar Heliothis armigera larvae were infected with a granulosis and a nuclear polyhedrosis virus, and all the externally visible symptoms for each virus are described. The effects of the virus infections on the feeding habits of the insects are also described, and it was found that a granulosis infection can prolong the larval period by up to 100%. The larvae continue feeding during this prolonged larval period, and can reach almost double the size and mass of normal larvae.It was further found that each of the viruses displays a distinct set of symptoms which could indicate beyond any doubt which of the two viruses induced death in the host.     

β-fructosidase activity in the silk glands of Heliothis zea

Extracts from the silk glands of Heliothis zea readily hydrolyzed carbohydrates in which the monosaccharides are linked by a β-fructosidic bond, i.e. sucrose, raffinose, and inulin. They did not hydrolyzed other carbohydrates exhibiting other types of linkages. Thus the glands must possess a β-fructosidase. A pH 6.5 and a temperature of 37°C provided optimum conditions for enzyme activity. At these conditions, the Km for the enzyme was 355 mM (sucrose). The majority of the activity occurred when the larvae were 8, 9 and 10 days old.     

Cardiac responses to sensory stimulation in the adult tobacco budworm moth, Heliothis virescens

Electrocardiogram recordings were performed on intact Heliothis virescens moths using Ag-AgCl electrodes positioned on the body cuticular surface. Regular heart activity and its changes in response to mechanical and olfactory stimulations were analysed. Moths were also tested in their anemotactic orientation in response to olfactory stimulation. Results show that regular cardiac activity in Heliothis is cyclical and consists of the alternation of a high and a low spike-frequency period. This activity pattern is strongly influenced by sensory stimulation. Both mechanical stimulations at various intensities and olfactory stimulations with sex pheromone and 1-hexanol evoked tachycardiac effects when applied during low spike-frequency cardiac activity. In contrast, they did not affect high spike-frequency cardiac activity. It is concluded that the cardiac response in Heliothis is an effective indicator of sensory reception. It would therefore appear to represent a valid tool for testing insect reactivity. Accepted: 30 August 1997     

Fat body stimulation of ecdysone synthesis in Heliothis zea

Prothoracic glands of Heliothis zea pupae require both a humoral factor and prothoracicotropic hormone (PTTH) to synthesize ecdysone. The humoral factor is absent when pupae are maintained at diapause-sustaining temperatures. Thus, pupae remain in diapause despite the release of PTTH at or before larval-pupal ecdysis.Tissue implantation experiments revealed that a diapause-terminating factor is present in the fat body of non-diapausing pupae. Other tissue implantation experiments showed that, when diapausing pupae were transferred from 19 to 27°C, diapause-terminating activity appeared first in the fat body and then the fat body into the haemolymph. HPLC separation of the haemolymph and fat body fractions followed by bioassay demonstrated that fractions containing diapause-terminating activity eluted from both tissues within 28–30 min. These results suggest that the factors found in the fat body and haemolymph may be the same compound.Evidence from ecdysone radioimmunoassay experiments ruled out the possibility that the diapauseterminating activity was due to either free or conjugated ecdysteroids. Corresponding in vitro experiments in which the prothoracic glands were cultured with brain extracts versus fat body and haemolymph fractions also indicated that the haemolymph/fat body factor was not PTTH.     

Establishment of early instar Heliothis zea on soybeans

Early instar Heliothis zea (Boddie) (Lepidoptera: Noctuidae) established on all stages of soybean (Glycine max [L.] Merrill) tested when larvae were confined to plants, although establishment decreased after podset stage and as seeds matured. In tests where larvae were allowed to disperse freely on plants, establishment also decreased as maturity increased, but was much lower in each stage than when confined to plants. Neonate larvae settled more quickly on terminals and expanding trifoliates than on mature trifoliates based on initiation of feeding and movement away from initial sites of placement. The ability of neonate larvae placed a short distance from soybean plants to reach a host was affected by high soil and ambient temperature. The results of these experiments suggest that larval host establishment and spin-down behavior play a major role in the late instar population distribution among soybean fields and that maturity of the host strongly influences that behavior.

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Résumé Des expériences ont été réalisées dans deux types de conditions (avec ou sans possibilité de dispersion) pour déterminer si le taux d'installation des chenilles de premier stade de H. zea Bod. dépendait d'une mortalité liée au stade développement de Glycine max L. L'installation était considérée comme réussie quand les chenilles avaient atteint le second stade. Bien qu'il fût possible de distinguer les variétés de soja tant par le taux d'installation des chenilles que par les dégâts provoqués, les plantes les plus jeunes étaient plus colonisées par des chenilles que les témoins plus âgés. Toutes les variétés de soja à n'importe quel stade étaient acceptées lorsque la dispersion des chenilles était impossible. Avec possibilité de dispersion, les chenilles ont aussi accepté tous les stades phénologiques, mais en moins grands nombres que lorsque la dispersion était impossible. Le comportement de dispersion était beaucoup plus fréquent sur les plantes à port indéterminé que sur celles à port déterminé. Les possibilités de réinstallation après dispersion des chenilles de premier stade ont été limitées.

     

Temporal oviposition patterns of Heliothis zea and Spodoptera ornithogalli

Egg production under laboratory conditions was examined over the lifespan of Heliothis zea (Boddie) and Spodoptera ornithogalli Guenée (Lepidoptera: Noctuidae). Although H. zea oviposits singly and S. ornithogalli oviposits in masses, temporal trends were similar. Egg numbers peaked shortly after mating and then rapidly declined. Egg weights also peaked shortly after mating, but decreased gradually over time. Temporal oviposition patterns were more erratic for unmated than mated females, suggesting the importance of mating in establishing the shape of the oviposition curve.

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Résumé La production d'ufs d'Heliothis zea (Boddie) et de Spodoptera ornithogalli Guenée (Lepidoptera: Noctuidae) durant toute la vie adulte a été examinée au laboratoire. Bien que H. zea dépose ses ufs isolément et que S. ornithogalli les dépose en groupes, les courbes temporelles d'oviposition chez des individus fécondés sont similaires. Le nomber d'ufs culmine peu après l'accouplement, puis décline rapidement. Le poids des ufs culmine aussi peu après l'accouplement, puis décroît graduellement. Des femelles fécondées produisent environ deux fois autant d'ufs que les femelles non fécondées, quoique la longévité ne diffère pas significativement entre les deux groupes. Des femelles non fécondées montrent des distributions temporelles plus irrégulières que des femelles fécondées, en ce qui concerne le nombre des ufs et leur poids. Ces irrégularités ont été attribuées à la tendance des femelles non fécondées à retenir leur ufs, ce qui suggère que l'accouplement exerce une influence sur la forme de la courbe temporelle d'oviposition. Des femelles d'H. zea contenant un, deux ou trois spermatophores n'ont pas produit des nombres d'ufs significativement différents.

     

Hormonal control of egg maturation in the corn earworm, Heliothis zea

At eclosion, the ovaries of female Corn earworm Heliothis zea do not contain mature eggs. Virgin-unfed females produced approximately 400 mature eggs in 8 days; mating or feeding doubled this number, and mating plus feeding more than tripled it. Females allatectomized or decapitated at day O matured few eggs. Egg production was restored by implantation of active corpora allata (CA) or by treatment with the juvenile hormone (JH) analogue methoprene at day 0. 20-Hydroxyecdysone, on the other hand, had no effect. Females in which the CA had been denervated or in which the median neurosecretory cells of the brain had been ablated at day O produced fewer eggs than sham-operated animals. These results indicate that egg maturation is controlled by JH and that continuous input from the brain is required for sustained CA activity for maintaining a high rates of egg maturation.The rate of JH biosynthesis by CA in vitro was determined with a radiochemical assay. The major hormones produced were JH-II and JH-III with small quantities of JH-I. The rates of JH synthesis were similar in all experimental groups which may indicate that the in vitro rate of JH synthesis does not reflect the actual state of CA activity in the female.     

Production of a monoclonal antibody to the arylphorin of Heliothis zea

A species-specific monoclonal antibody was produced to whole plasma of fifth instar larvae of the corn earworm, Heliothis zea (Boddie) (Lepidoptera:Noctuidae). This antibody did not cross-react with proteins from the plasma of any of the other lepidopteran larvae tested, including other Heliothis spp. The antigen recognized by this antibody was characterized and found to be arylphorin, a prominent larval storage protein. This conclusion was based on the electrophoretic as well as immunological characteristics of the antigen. Polyacrylamide gel electrophoresis indicated that the antigen had an apparent native molecular weight of 460,000, and that it was composed of subunits having apparent molecular weights of 76,000. The isoelectric point of the antigen was 5.9, with some microheterogeneity being seen. Western blotting of arylphorin against the monoclonal antibody clearly identified the antigen as arylphorin. This protein was not found in egg homogenates or early instar larval plasma, but it was present in large quantities in pupal homogenates and, at trace levels, in adult homogenates. The efficient production and selection of hybridomas producing antibodies specific to H. zea arylphorin using unfractionated plasma as immunogen illustrates the fact that monoclonal antibody technology can produce highly specific antibodies using crude antigen preparations. We discuss the tradeoffs one must accept when choosing this strategy over one using purified immunogens.     

Aldehyde-oxidizing enzymes in an adult moth: in vitro study of aldehyde metabolism in Heliothis virescens

The conversion of pheromonal aldehydes to carboxylic acids in vitro in tissue extracts of Heliothis virescens is catalyzed by both aldehyde dehydrogenase and aldehyde oxidase enzymes. The aldehyde-oxidizing activity in antennae, heads, legs, and hemolymph from male and female moths was examined by radiochromatographic and spectroscopic assays. First, the enzymatic activity was measured in the presence or absence of added NAD+ using either (Z)-9-tetradecenal or (Z)-11-hexadecenal as tritiated substrate. Second, substrate specificity was determined spectroscopically by (i) indirect measurement of the AO-released hydrogen peroxide through the coupled AO-horseradish peroxidase reaction and by (ii) direct measurement of the ALDH-produced NADH. Both aldehyde-oxidizing activities were associated with soluble enzymes in the antennal extracts, and these enzymes degraded pheromone and nonpheromonal aldehydes. Both AO and ALDH activities were present in male and female tissues. AO activity was exhibited primarily in the antennal extracts and to a lesser degree in the leg extracts. Moreover, ALDH activity was distributed in the antenna, head, and leg extracts. A vinyl ketone analog of (Z)-11-hexadecenal preferentially inhibited the ALDH activity over the AO activity.     

Replication of Heliothis zea nuclear polyhedrosis virus in cloned cell lines

Clones from two Heliothis zea (Lepidoptera:Noctuidae) ovarian cell lines, BCIRL-Hz-AM1 (AM1) and BCIRL-Hz-AM3 (AM3), were generated and their ability to produce H. zea nuclear polyhedrosis virus (HzSNPV) was compared. Titers of extracellular virus (ECV) ranged from 5.5 (AM3-F9) to 44.9 x 10(5) PFU/ml (AM1-A4), with the parental cell lines AM3 and AM1 producing 14.8 and 26.4 x 10(5) PFU/ml, respectively. Concentrations of polyhedral inclusion bodies (PIB) produced by the cloned lines ranged from 0.7 (AM3-F9) to 59.6 x 10(6) PIB/ml (AM1-B3), while the parental cell lines generated 6.5 (AM3) and 12.9 x 10(6) PIB/ml (AM1). The percentage of cells from the cloned lines that produced PIB ranged from 39 to 86.4% for AM3-F9 and AM1-A7, respectively, with the parental lines exhibiting 49.1% (AM3) and 75.3% (AM1) cells with PIB. The number of PIB per cell also differed markedly between cell lines, varying from 18.3 (AM3-F9) to 184.4 (AM1-B3) PIB/cell. The parental lines produced 57.3 (AM3) and 75.9 (AM1) PIB/cell. Thus, significant differences were seen in virus production (ECV, PIB) between parental cell lines, as well as between parental cell lines and their clones. In addition, cell lines were characterized with regard to their growth rates and isoenzyme patterns.     

Replication and serial passage of infectious Heliothis nucleopolyhedrosis virus in an established line of Heliothis zea cells

An established cell line derived from the ovary of adults of the cotton bollworm, Heliothis zea, supported growth of the Heliothis nucleopolyhedrosis virus (NPV). Typical NPV symptoms were obtained when infected cells were fed to neonatal bollworms; however, the cell line never produced free virions or inclusion bodies containing virions. Infectious virus was passed through the cell line 7 consecutive times, using only infected cells from the previous pass. Infectivity at the 7th serial-pass represented a dilution of >10⁻⁸ of the original inoculum.     

The role of the frontal ganglion and corpora cardiaca/corpora allata complex in post-feeding weight loss in adult Heliothis zea

Abstract A post-feeding diuretic response has been observed in adult Heliothis zea (Boddie) (Lepidoptera: Noctuidae). 2-day-old starved adults which were allowed to feed to repletion on a 10% (w/v) sucrose solution lost 49.1% (females) and 85.8% (males) of the weight of the ingested meal during the first hour following feeding. Ligation between head and thorax or frontal ganglionectomy, when performed immediately following feeding, each resulted in a significant and permanent reduction in this normal weight-loss. Injection of homogenates of the corpora cardiaca/corpora allata (CC/CA) complex into non-ligated insects immediately after feeding also reduced significantly the post-feeding weight-loss, but this inhibition was transient and disappeared after 1 h. Dissection and weighing of the crop from either ligated, frontal ganglionectomized, or CC/CA-injected insects confirmed the crop as the predominant site of fluid retention in each case. We suggest that a soluble antidiuretic factor from the CC/CA acts in conjunction with the frontal ganglion to control the rate of crop emptying and subsequent diuresis by regulating the volume of ingested fluid that is passed into the haemolymph from the crop/midgut.     

Natural diets and self-selection: Heliothis zea larvae and maize

Although the first two instars tend to eat the silks, last instar larvae of the corn earworm, Heliothis zea (Boddie) (Lepidoptera: Noctuidae), prefer to feed on the kernels rather than on the other components of the maize ear. They feed preferentially on the germ of the kernel, but when offered whole kernels also eat some of the endosperm. Larvae that eat cut portions of the kernels that include all of the germ and some endosperm utilize their food for growth more efficiently than do larvae that eat portions of the kernel that include only endosperm. Adults that ate germ portions or endosperm portions of the kernel as larvae do not differ significantly in longevity, number of days on which they oviposit, number of eggs laid per day or the percentage of the eggs that hatched. We suggest that the larvae use self-selection to increase their intake of germ, thus increasing utilization efficiency and decreasing the amount of food that they must eat. When they feed on plants (other than maize) with small fruits, this probably increases their ability to compete for food when it is scarce and minimizes exposure to predation by decreasing the number of foraging trips and the distance that they must travel in search of food.

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Résumé Bien que les deux premiers stades larvaires d'H. zea (Lepid.: Noctuidae) tendent à consommer les soies, les trois derniers stades préfèrent les grains aux autres parties de l'épi de maïs. La chenille du dernier stade consomme de préférence le germe du grain, mais quand on lui offre l'ensemble du grain, elle consomme aussi une partie de l'albumen. Les chenilles qui coupent des morceaux du grain comprenant la totalité du germe et une fraction d'albumen ont une alimentation permettant une croissance plus efficace que les chenilles qui consomment des portions de grain ne contenant que de l'albumen. Ainsi, les chenilles qui mangent des fractions de germe ont besoin de consommer moins, pour atteindre le même poids que les chenilles qui consomment des morceaux d'albumen. Les longévités, les nombres de jours de ponte, les fécondités quotidiennes et les fertilités des oeufs sont les mêmes pour les papillons provenant des chenilles ayant consomme des morceaux de germe ou des morceaux d'albumien. L'albumen seul est un aliment suboptimal, et H. zea pourrait théoriquement améliorer ultérieurement son régime en mangeant plus de germes. Nous estimons que ceci ne serait pas rentable puisque la durée des stades devrait être prolongée et que la manutention et les rejets d'albumen devraient être accrus. H. zea se serait adapté à une solution de compromis pour l'optimalisation du régime alimentaire. Il réduit le temps de récolte et perd en efficacité en consommant de l'albumin suboptimal; mais il regagne autant que possible en efficacité en mangenat le germe de pratiquement tous les grains attaqués. Cette augmentation de l'efficacité diminue la quantité d'aliments qui doit être ingéré, et fournit un potentiel adaptatif quand les chenilles consomment des plantes (autres que le maïs) avec des petits fruits. L'exposition à la prédation est réduite, puisque la distance parcourue pour la recherche des fruits est diminuée.

     

Vitellogenin and egg production in the moth,Heliothis virescens

The characteristics of vitellogenin (Vg) and the relationship between Vg production and egg production in the tobacco budworm, Heliothis virescens, were studied. The relationship between Vg production and juvenile hormone (JH) and the impact of mating on Vg and egg production were also investigated. Vg appears in the hemolymph of H. virescens about 6 h after moth eclosion. Vg may be separated into two apoproteins (ApoVg-I and ApoVg-II) by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The molecular weights were calculated to be 156,065 ± 800 for ApoVg-I and 39,887 ± 323 for ApoVg-II. SDS-PAGE analysis revealed that the female hemolymph Vg polypeptides appear to be identical to those from eggs but are absent in male hemolymph. Vg concentration was significantly higher in mated females than in virgin females of the same age at 48 h after emergence. Rates of egg production increased as Vg production increased; rates of egg production in mated females were significantly higher than those of virgin females at 48, 72, 96, and 120 h postemergence. Vg production is dependent on JH, because hemolymph from decapitated females lacked Vg while that of decapitated females treated with synthetic JH had Vg at levels comparable to similarly aged, normal H. virescens females. Hemolymph JH titers in mated females were significantly higher compared with those in virgin females at all sampling periods. The high JH level in mated females may explain the high Vg and egg production in mated H. virescens. Arch. Insect Biochem. Physiol. 34:287–300, 1997. © 1997 Wiley-Liss, Inc.