Regulation of larval diapause by juvenile hormone in the European corn borer,Ostrinia nubilalis

The effects of juvenile hormone (JH) on the larval diapause of the European corn borer, Ostrinia nubilalis, were investigated. The larval period was prolonged and the development of gonads was retarded if non-diapause larvae were treated with JH. Pupation of diapausing larvae was accelerated by allatectomy. It was concluded that the diapause of this borer is largely regulated by JH.     

Elevated temperature increases reproductive investment in less preferred mates in the invasive European corn borer moth

Rapidly changing environments may weaken sexual selection and lead to indiscriminate mating by interfering with the reception of mating signals or by increasing the costs associated with mate choice. If temperature alters sexual selection, it may impact population response and adaptation to climate change. Here, we examine how differences in temperature of the mating environment influence reproductive investment in the European corn borer moth (Ostrinia nubilalis). Mate preference in this species is known to be related to pheromone usage, with assortative mating occurring between genetically distinct E and Z strains that differ in the composition of female and male pheromones. We compared egg production within and between corn borer lines derived from four different populations that vary in pheromone composition and other traits. Pairs of adults were placed in a mating environment that matched the pupal environment (ambient temperature) or at elevated temperature (5°C above the pupal environment). At ambient temperature, we found that within‐line pairs produced eggs sooner and produced more egg clusters than between‐line pairs. However, at elevated temperature, between‐line pairs produced the same number of egg clusters as within‐line pairs. These results suggest that elevated temperature increased investment in matings with typically less preferred, between‐line mates. This increased investment could result in changes in gene flow among corn borer populations in warming environments.     

Changes in calling behaviour and mating success in the European corn borer (Ostrinia nubilalis), caused by relative humidity

We examined the calling behaviour of O. nubilalis virgin females, held at three different constant conditions of relative humidity from the time of emergence through their sixth night of calling, as well as the incidence of mating over time at low or high relative humidity conditions. The mean age at which females initiated calling in their life was independent of relative humidity conditions, although more variable at low humidity. On successive nights of calling, fewer virgin European corn borer females called under low compared with high humidity conditions, and those that did began later in scotophase, had fewer calling bouts, and spent less time calling. These changes in the female prereproductive behaviour were reflected on the incidence of mating which, in the first two days following emergence, was significantly less at low than at high humidity conditions. The ecological significance of these results is discussed in relation with the susceptibility of O. nubilalis to desiccation.

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Résumé Nous avons étudié le comportement d'appel des femelles vierges d'Ostrinia nubilalis maintenues sous trois conditions constantes d'humidité relative, depuis l'émergence jusqu'à la sixième nuit d'appel, ainsi que l'incidence d'accouplement d'individus placés en milieu sec ou humide pour diverses périodes de temps. Les conditions d'humidité n'ont pas significativement affecté l'âge moyen auquel les femelles ont commencé à appeler au cours de leur vie, bien qu'il ait été plus variable à 53% h.r. Les nuits subséquentes, le comportement d'appel des femelles maintenues à basse humidité a débuté plus tard dans la nuit, s'est exprimé pour une période de temps plus courte avec moins de séquences d'appel que celui de leurs congénères placés en milieu humide, et la proportion de femelles en appel a toujours été plus faible à 53% h.r. Ces changements du comportement préreproducteur des femelles se sont répercutés sur l'incidence d'accouplement qui, les deux premiers jours suivant lémergence, a été significativement plus faible à basse humidité qu'elle ne l'a été en milieu humide. Ces résultats seront discutés en relation avec l'écologie de O. nubilalis et associés à sa susceptibilité à la dessication.

     

Concerted evolution of male and female display traits in the European corn borer, Ostrinia nubilalis

Background  

Sexual reproduction entails the encounter of the sexes and the multiplicity of rituals is parallel to the diversity of mating systems. Evolutionary mechanisms such as sexual selection and sexual conflict have led to the elaboration of traits to gain attention and favours from potential partners. A paradox exists about how coordinated systems can evolve and diverge when there would seem to be a stabilising selection acting. Moth display traits – pheromones – constitute an advantageous model with which to address questions about the evolution of mating systems in animals. Both males and females can possess pheromones that are involved either in close- or long-range communication. Female and male pheromones appear to have different origins and to be under different evolutionary constraints, thus they might be envisioned as independently evolving traits. We conducted laboratory experiments to explore the role of scents released during courtship by males of the European corn borer, Ostrinia nubilalis.     

Multiple mating and lifetime reproductive output in female dobsonflies that receive nuptial gifts

In mating of the dobsonfly, Protohermes grandis (Thunberg), the male attaches the spermatophore externally to the female genitalia. The spermatophore includes a large gelatinous mass which the female detaches and feeds on after mating. While the female consumes this nuptial food gift, sperm is evacuated from the remaining portion of the spermatophore (sperm package) into her reproductive tract. Under laboratory conditions, mated females maintained receptivity throughout their lifetime, and they remated even on the day following copulation. A single insemination may supply enough sperm, as females mated only once deposited fertile eggs throughout life and, when dissected after death, all females had sperm in the spermatheca. There was a positive correlation between longevity and the number of matings. Lifetime fecundity also increased as mating multiplied. However, the size of eggs and hatchlings was not influenced by the number of matings. It seems that large spermatophore consumption by female P. grandis provides nutrients that increase fitness not in offspring quality, but in their quantity.     

The role of juvenile hormone in the larval diapause of the European corn borer,Ostrinia nubilalis

The possible role of juvenile hormone (JH) in the induction and termination of larval diapause in the European corn borer, Ostrinia nubilalis, was investigated using topical applications of both JH I and a JH mimic as well as by monitoring JH titers with the Galleria bioassay. Neither JH nor the JH mimic ZR515 was capable of influencing diapause termination when administered topically. The Galleria bioassay revealed little or no JH in the hemolymph of mid diapause (>30 days) insects, indicating no demonstrable role for JH in diapause maintenance. When ZR515 was administered to nondiapause, newly ecdysed fifth instar larvae the pupal molting cycle was delayed. By use of photoperiodic regimes we were able to show that the molting delay was not equivalent to diapause induction. The Galleria bioassay showed differences in JH titer profiles between diapause and nondiapause animals during the final larval stadium. The nondiapause insects showed titers that decline rapidly to trace amounts following the molt to fifth instar then rose prior to pupation. The diapause insects had generally higher titers and exhibited a more gradual decline after the molt. No evidence was obtained to support the hypothesis that JH plays a key role in the induction, maintenance, or termination of larval diapause.     

Regulation of pheromone biosynthesis in the "Z strain" of the European corn borer, Ostrinia nubilalis

The regulation of pheromone biosynthesis by the neuropeptide PBAN in the Z strain of the European corn borer, Ostrinia nubilalis, was investigated using labeled intermediates. Injection of radiolabeled acetate showed PBAN did not influence the de novo synthesis of saturated fatty acids in the gland. When deuterium-labeled myristic acid was topically applied to the gland, females injected with PBAN produced more labeled pheromone than did control females, indicating that PBAN controls one of the later steps of pheromone biosynthesis. Although more myristic acid was Delta11-desaturated in the gland in the presence of PBAN, this was counterbalanced by less Delta11-desaturation of palmitic acid, indicating that desaturase activity did not change overall. This change in flux of myristic acid through to pheromone was shown to be caused by increased reduction of fatty acid pheromone precursors occurring in the presence of PBAN.     

Male indifference to female traits in an arctiid moth (Utetheisa ornatrix)

Abstract.  1. Female Utetheisa ornatrix (Lepidoptera: Arctiidae) mate selectively with large males able to transmit sizeable quantities of nutrient and defensive pyrrolizidine alkaloid with the spermatophore. The female gauges male size indirectly by assessment of the male's courtship pheromone.
2. Male Utetheisa invest upward of 10% of body mass in spermatophore production, and could therefore have been expected to be choosy; however, when offered females differing in alkaloid content, body mass, or mating status, males showed disregard of these parameters, both in their choice of partner and in their allocation of resources to the spermatophore.
3. It is concluded that Utetheisa males do not have the option to select females by comparison shopping . Females broadcast their attractant pheromone for less than an hour per day. Given this time constraint and the potentially high cost of mate localisation, males may have no choice but to mate on an opportunistic basis.     

Mating frequency and reproductive success in an income breeding moth,Mnesampela privata

The frequency of mating in insects is often an important determinant of female reproductive output and male sperm competition. In Lepidoptera that provide male nutrients to the female when mating, it is hypothesized that polyandry may be more prevalent. This is thought to be especially so among species described as income breeders; that is, in species who do not derive all their nutrients for reproductive output entirely from the resources obtained during the larval stage. We selected the geometrid moth, Mnesampela privata (Guenée) (Lepidoptera: Geometridae), to examine this hypothesis further. We found this species was best characterized as an income breeder with female weight on emergence positively correlated with total egg load but not with the number of eggs laid. Further, in accord with income breeders, females emerged with a partially developed egg load and lifetime fecundity was positively correlated with the number of oviposition days. However, in the laboratory we found that incidence of repeated matings or polyandry was rare. When moths were paired singly over their lifetime, only 4% of mated females multiple mated. When females were paired with three males concurrently, female mating success increased from 60 to 81% with multiple mating among mated females increasing to just 15%. Dissection of wild caught M. privata found that polyandry levels were also low with a maximum of 16.4% of females collected at any one time being multiple mated. In accord with theory, mating significantly increased the longevity of females, but not of males, suggesting that females acquire essential resources from male ejaculates. Despite this, multiple mated females showed a trend toward decreasing rather than increasing female reproductive output. Spermatophore size, measured on death of the female, was not correlated with male or female forewing length but was negatively correlated with the number of fertile eggs laid and female longevity. Smaller spermatophore width may be related to uptake of more nutrients by the female from a spermatophore. We discuss our findings in relation to income breeding and its relationship to polyandry in Lepidoptera.     

Evaluating female remating rates in light of spermatophore degradation in Heliconius butterflies: pupal‐mating monandry versus adult‐mating polyandry

1. Butterflies are frequently used in comparative studies of sexual selection because of their diverse mating systems. In Heliconius, the two major clades in the genus are characterised by contrasting pupal‐mating and adult‐mating strategies. Adult‐mating females are considered to be promiscuous whereas pupal‐mating females are thought to be monandrous. 2. Counting spermatophores in female Lepidoptera is a common method for assessing patterns of female remating. However, in pupal‐mating Heliconius butterflies spermatophores can become completely degraded, potentially leading to underestimation of female remating rates. 3. We qualitatively characterised the different states of spermatophore degradation, and showed that complete degradation takes approximately 3 weeks in captive‐bred H. erato females. 4. We counted spermatophores and/or assayed spermatophore degradation in > 500 Heliconius females across 28 species sampled from natural populations. Among pupal‐maters these observations yielded a few rare observations of double mating by recently eclosed females, but generally indicated a lack of rematings. In contrast, approximately 25% of sampled adult‐mating females remated at least once. 5. Using a novel statistical analysis, we estimated remating rates from patterns of spermatophore degradation or from counts stratified by age, as indicated by wing‐wear. This analysis showed no statistically significant evidence for remating for the pupal‐mating H. erato whereas significant remating rates were detected for adult‐mating species. 6. The present results support the established view of Heliconius mating systems in which pupal‐maters are largely monandrous, whereas adult‐maters are polyandrous.     

Dynamics of Nosema pyrausta in natural populations of the European corn borer, Ostrinia nubilalis: A six-year study

Nosema pyrausta (Paillot) (Microsporida: Nosematidae) is an obligatory intracellular parasite of the European corn borer, Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae). This pathogen is maintained in natural populations of O. nubilalis by both horizontal and vertical transmission. The impact of N. pyrausta on fecundity of adults and survival of larvae has been well documented in laboratory and field research. In an extensive study covering a 6 year period at one site, we described the effect of N. pyrausta within O. nubilalis populations in a continuous corn following corn ecosystem. We documented the presence of the pathogen through all life stages of O. nubilalis (egg, larva, pupa, adult), by collecting throughout the crop season and examining each insect stage in the laboratory for the frequency of infection with N. pyrausta. The percentage of infection with N. pyrausta and magnitude of the O. nubilalis population fluctuated throughout generation 1 and generation 2. Both horizontal and vertical transmission played a role in maintaining N. pyrausta in the population in both generations. There were strong correlations between percentage adults with N. pyrausta and percentage larvae with N. pyrausta, and between percentage eggs with N. pyrausta and percentage larvae with N. pyrausta. There was a weak correlation between percentage adults with N. pyrausta and percentage eggs with N. pyrausta. The percentage of insects infected with N. pyrausta was always lowest in the egg.     

Dissemination of Nosema pyrausta in feral populations of the European corn borer,Ostrinia nubilalis

Epizootiological studies of Nosema pyrausta in natural European corn borer populations show that while vertical transmission is the primary way in which N. pyrausta is transferred from one host generation to the next, it is horizontal transmission that is responsible for the annual build-up of infection in each nonoverlapping generation. During the first generation, larval corn borer migration to adjacent corn stalks is minimal and increases in the prevalence of N. pyrausta within the population result from horizontal transmission of infection among borers that inhabit the same stalk. During the second generation, corn borer larvae actively disperse to other corn plants and this results in an increased level of infection. Factors which facilitate pathogen dispersal in this generation include (1) higher host densities, (2) longer periods of larval development, (3) lower mortality among young larvae, and (4) possible mechanical transmission by the braconid parasitoid, Macrocentrus grandii.     

Distribution of putative neurotransmitters in the brain of the European corn borer, Ostrinia nubilalis.

Radioautographic localization of three putative neurotransmitters in the brain of larvae of the European corn borer was studied by electron microscopy. Acetylcholine and dopamine were found to be accumulated in median neurosecretory cells. Both acetylcholine and dopamine were preferentially found in glial cells as compared to neuronal cells. This may be indicative of active participation of glial cells in a blood-brain barrier in the intact brain. Gamma-aminobutyric acid was not accumulated by any particular cell in the brain, but was apparently excluded to the periphery of the ganglion.It is suggested that dopamine, or more likely an immediate metabolite may be involved in an endogenous time measuring system and/or diapause induction-termination.     

Reduction of damage by the Mediterranean corn borer, Sesamia nonagrioides, and the European corn borer, Ostrinia nubilalis, in maize fields by a trifluoromethyl ketone pheromone analog

Large‐scale field experiments on the Mediterranean corn borer, Sesamia nonagrioides Lefèbvre (Lepidoptera: Noctuidae), were carried out in 2004–2006 on maize [Zea mays L. (Poaceae)] fields using (Z)‐11‐hexadecenyl trifluoromethyl ketone, an antagonist analog of the pheromone of this species, to evaluate a possible reduction of damage caused by this pest. The effect of the treatments on the European corn borer, Ostrinia nubilalis Hübner (Lepidoptera: Crambidae), a sympatric species, was also determined. Evaluation of the success of the experiments was assessed by counting (i) the number of males caught by pheromone traps in treated and untreated fields, (ii) the number of plants attacked by both insects in both plots, and (iii) the number of larvae present in infested plants in both fields. Effectiveness of the treatment was high for the second generation of the Mediterranean corn borer, the most harmful to the crop (86–90% reduction in the number of plants attacked and 67–98% reduction in the number of larvae per plant in treated fields in comparison to untreated fields), and moderate for the third generation (reduction of 41–71% and 33–77%, respectively). Treatments were also effective for the second generation of the European corn borer (61–75% reduction in the number of plants attacked, 58–78% reduction in the number of larvae found per plant) as well as for the third generation (69–97% and 70–98% reduction, respectively). By plotting the amount of the antagonist remaining on the dispensers after 40–45 days of exposure with time, the mean release rate of the compound was calculated to be 2.2%/day in 2004, 1.95%/day in 2005, and 2.1%/day in 2006, with 26% of the initial compound remaining after 20 days of experimentation. The emission rate appears to cover the flight of the most damaging second generation of both insects. Prospects of using trifluoromethyl ketones as new potential agents for pest control are also outlined.     

Effectiveness of Trichogramma ostriniae and T. nubilale for controlling the European corn borer Ostrinia nubilalis in sweet corn

Seven field releases of Trichogramma ostriniae and T. nubilale (Hymenoptera: Trichogrammatidae) were made separately and in combination in a sweet corn field to compare the level of parasitism in sentinel eggs of the European corn borer, Ostrinia nubilalis (Lepidoptera: Pyralidae). The results indicate that the levels of egg parasitism among different release dates differed mainly because of changes in weather and plant architecture within the season. The level of egg parasitism by releasing T. ostriniae alone was found to be 15% higher than that by releasing T. nubilale alone, and 20% higher than by releasing the combination of the two species. Further analyses using the logistic regression model for independent and correlated data indicated T. ostriniae to be more efficient at discovering host egg masses and to have higher levels of egg parasitism than T. nubilale. Mutual interference between T. ostriniae and T. nubilale was the main factor for the lower level of egg parasitism when T. ostriniae and T. nubilale were released together. The results suggest that T. ostriniae is the better candidate for augmentative releases for control of the European corn borer, and the two species should not be released into a corn field at the same time.     

Distribution of digestive proteinases in the alimentary tract of the european corn borer Ostrinia nubilalis (lepidoptera: Pyralidae)

The distribution of digestive proteinases in either the anterior and posterior midgut or between the midgut epithelium and ectoperitrophic and endo-peritrophic spaces in the midgut were examined in the European corn borer, Ostrinia nubilalis. Trypsin, chymotrypsin, elastase, and aminopeptidase activities were the same in the anterior and posterior halves of the midgut. Of the total aminopeptidase activity, 95% was located in the midgut epithelium, and 90% of the trypsin, 97% of chymotrypsin, and 93% of the elastase activity were found in the midgut lumen. Trypsin, measured by hydrolysis of benzoyl-L-arginine ethyl ester, and chymotrypsin levels were significantly higher in the ectoperitrophic space compared to the endoperitrophic space. Digestion in the midgut is proposed to be sequential with tryptic digestion occurring in the endoperitrophic space. Ingested protein is digested further in the ectoperitrophic space by the action of elastase, chymotrypsin, and a second trypsin. Final digestion occurs by an intracellular aminopeptidase. © 1995 Wiley-Liss, Inc.     

Microbial diversity in the midguts of field and lab-reared populations of the European corn borer Ostrinia nubilalis

BACKGROUND: Insects are associated with microorganisms that contribute to the digestion and processing of nutrients. The European Corn Borer (ECB) is a moth present world-wide, causing severe economical damage as a pest on corn and other crops. In the present work, we give a detailed view of the complexity of the microorganisms forming the ECB midgut microbiota with the objective of comparing the biodiversity of the midgut-associated microbiota and explore their potential as a source of genes and enzymes with biotechnological applications. METHODOLOGICAL/PRINCIPAL FINDINGS: A high-throughput sequencing approach has been used to identify bacterial species, genes and metabolic pathways, particularly those involved in plant-matter degradation, in two different ECB populations (field-collected vs. lab-reared population with artificial diet). Analysis of the resulting sequences revealed the massive presence of Staphylococcus warneri and Weissella paramesenteroides in the lab-reared sample. This enabled us to reconstruct both genomes almost completely. Despite the apparently low diversity, 208 different genera were detected in the sample, although most of them at very low frequency. By contrast, the natural population exhibited an even higher taxonomic diversity along with a wider array of cellulolytic enzyme families. However, in spite of the differences in relative abundance of major taxonomic groups, not only did both metagenomes share a similar functional profile but also a similar distribution of non-redundant genes in different functional categories. CONCLUSIONS/SIGNIFICANCE: Our results reveal a highly diverse pool of bacterial species in both O. nubilalis populations, with major differences: The lab-reared sample is rich in gram-positive species (two of which have almost fully sequenced genomes) while the field sample harbors mainly gram-negative species and has a larger set of cellulolytic enzymes. We have found a clear relationship between the diet and the midgut microbiota, which reveals the selection pressure of food on the community of intestinal bacteria.     

Resource availability,mating opportunity and sexual selection intensity influence the expression of male alternative reproductive tactics

The expression of alternative reproductive tactics can be plastic and occur simultaneously depending on cues that vary spatially or temporally. For example, variation in resources and sexual selection intensity is expected to influence the pay‐off of each tactic and shape the decision of which tactic to employ. Males of the nuptial gift‐giving spider Pisaura mirabilis can adopt three tactics: offering a genuine prey gift, a ‘worthless’ non‐nutritious gift or no gift. We hypothesized that resources and/or male body condition, and mating opportunity and sexual selection intensity, vary over the course of the mating season to shape the co‐existence of alternative traits. We measured these variables in the field over two seasons, to investigate the predictions that as the mating season progresses, (i) males become more likely to employ a gift‐giving tactic, and (ii) the likelihood of switching from worthless to genuine gifts increases. Prey availability increased over the season and co‐varied with the propensity of males to employ the gift‐giving tactic, but we found no support for condition‐dependent gift giving. Males responded to an increase in female availability by increasing their mating effort (gift production). Furthermore, the frequency of genuine gift use increased with sexual selection intensity, consistent with the assumption that sperm competition intensity increases with time. Our results suggest that the frequency of alternative tactics is shaped by seasonal changes in ecological factors and sexual selection. This leads to relaxed selection for the gift‐giving tactic early in the season when females are less choosy and resources more scarce, and increased selection for genuine gifts later in the season driven by mating opportunity and risk of sperm competition.     

Sex-linked pheromone receptor genes of the European corn borer, Ostrinia nubilalis, are in tandem arrays

Background

Tuning of the olfactory system of male moths to conspecific female sex pheromones is crucial for correct species recognition; however, little is known about the genetic changes that drive speciation in this system. Moths of the genus Ostrinia are good models to elucidate this question, since significant differences in pheromone blends are observed within and among species. Odorant receptors (ORs) play a critical role in recognition of female sex pheromones; eight types of OR genes expressed in male antennae were previously reported in Ostrinia moths.

Methodology/Principal Findings

We screened an O. nubilalis bacterial artificial chromosome (BAC) library by PCR, and constructed three contigs from isolated clones containing the reported OR genes. Fluorescence in situ hybridization (FISH) analysis using these clones as probes demonstrated that the largest contig, which contained eight OR genes, was located on the Z chromosome; two others harboring two and one OR genes were found on two autosomes. Sequence determination of BAC clones revealed the Z-linked OR genes were closely related and tandemly arrayed; moreover, four of them shared 181-bp direct repeats spanning exon 7 and intron 7.

Conclusions/Significance

This is the first report of tandemly arrayed sex pheromone receptor genes in Lepidoptera. The localization of an OR gene cluster on the Z chromosome agrees with previous findings for a Z-linked locus responsible for O. nubilalis male behavioral response to sex pheromone. The 181-bp direct repeats might enhance gene duplications by unequal crossovers. An autosomal locus responsible for male response to sex pheromone in Heliothis virescens and H. subflexa was recently reported to contain at least four OR genes. Taken together, these findings support the hypothesis that generation of additional copies of OR genes can increase the potential for male moths to acquire altered specificity for pheromone components, and accordingly, facilitate differentiation of sex pheromones.