Molecular differentiation at nuclear loci in French host races of the European corn borer (Ostrinia nubilalis)

French populations of the European corn borer consist of two sympatric and genetically differentiated host races. As such, they are well suited to study processes that could be involved in sympatric speciation, but the initial conditions of host-race divergence need to be elucidated. Gene genealogies can provide insight into the processes involved in speciation. We used DNA sequences of four nuclear genes to (1) document the genetic structure of the two French host races previously delineated with allozyme markers, (2) find genes directly or indirectly involved in reproductive isolation between host races, and (3) estimate the time since divergence of the two taxa and see whether this estimate is compatible with this divergence being the result of a host shift onto maize after its introduction into Europe approximately 500 years ago. Gene genealogies revealed extensive shared polymorphism, but confirmed the previously observed genetic differentiation between the two host races. Significant departures from the predictions of neutral molecular evolution models were detected at three loci but were apparently unrelated to reproductive isolation between host races. Estimates of time since divergence between French host races varied from approximately 75,000 to approximately 150,000 years, suggesting that the two taxa diverged recently but probably long before the introduction of maize into Europe.     

Frequency of alleles conferring resistance to Bt maize in French and US corn belt populations of the European corn borer,Ostrinia nubilalis

Farmers, industry, governments and environmental groups agree that it would be useful to manage transgenic crops producing insecticidal proteins to delay the evolution of resistance in target pests. The main strategy proposed for delaying resistance to Bacillus thuringiensis ( Bt) toxins in transgenic crops is the high-dose/refuge strategy. This strategy is based on the unverified assumption that resistance alleles are initially rare (<10(-3)). We used an F(2) screen on >1,200 isofemale lines of Ostrinia nubilalis Hübner (Lepidoptera: Crambidae) collected in France and the US corn belt during 1999-2001. In none of the isofemale lines did we detect alleles conferring resistance to Bt maize producing the Cry1Ab toxin. A Bayesian analysis of the data indicates that the frequency of resistance alleles in France was <9.20 x 10(-4) with 95% probability, and a detection probability of >80%. In the northern US corn belt, the frequency of resistance to Bt maize was <4.23 x 10(-4) with 95% probability, and a detection probability of >90%. Only 95 lines have been screened from the southern US corn belt, so these data are still inconclusive. These results suggest that resistance is probably rare enough in France and the northern US corn belt for the high-dose plus refuge strategy to delay resistance to Bt maize.     

Observations on the seasonal incidence of microsporidiosis in European corn borer populations in Illinois

Summary Examinations of living and dead larvalPyrausta nubilalis (Hübner) collected in the field over a four-year period indicate that infections caused byPerezia pyraustae Paillot are widely distributed within populations ofP. nubilalis in Illinois. Observed fluctuations in the incidence of the parasite among living and dead insects collected in various seasons was taken as evidence that the disease interacts with temperature to cause mortality in populations of the host.

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Résumé Des examens des larves vivantes et mortes dePyrausta nubilalis (Hübner) collectées dans les champs pendant quatre années consécutives indiquent que des infections parPerezia pyraustae Paillot sont très fréquentes dans les populations deP. nubilalis en Illinois (états-Unis). Les fluctuations observées dans l'incidence du parasite dans les insectes vivants et morts recueillis pendant plusieurs saisons montrent que la maladie est influencée par la température, celle-ci occasionnant la mortalité dans les populations de l'h?te.

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An abridgment of a portion of a thesis submitted in partial fulfillment of the requirements for the degree Doctor of Philosophy in the Graduate College of the University of Illinois, 1958.     

Foliar application ofNosema pyrausta for suppression of populations of European corn borer

In 1974, an application of the microsporidan,Nosema pyrausta (Paillot, 1927)Kotlan, 1928, with a back-pack type sprayer (22.5×10⁷ spores/plant) to whorl stage maize infested with European corn borers,Ostrinia nubilalis (Hübner) reduced the number of larvae/plant by 48.1% and produced an infection of 15.3×10⁴ spores/mg of larval weight in 62.1% of the collected larvae. In 1975, applications of 24.3×10⁷ spores/plant to similar maize, in 2 separate tests, reduced the number of larvae/plant by 18.8 and 43.8% and caused an infection of 14.3 and 19.1×10⁴ spores/mg of larval weight in 65.9 and 63.3% of the collected larvae. Also, in 1975, applications of 24.3×10⁷ spores/plant to pollen shedding maize in 2 separate tests reduced the number of larvae/plant by 17.2 and 14.1% and caused an infection of 24.3 and 27.2×10⁴ spores/mg of larval weight in 99.2 and 95.2% of the collected larvae.     

Allozyme differentiation among nine populations of the corn borer (Ostrinia) in China

To test the hypothesis of the migration of the corn borer, the allozymes of nine populations of the corn borer(Ostrinia) in China were checked using verticalslab polyacrylamide gel electrophoresis. Eight loci of six allozymes were analyzed. The mean of the genetic identities among the nine populations calculated from the allele frequencies was 0.99068, much closer than that of other species and geographical populations. The mean (0.97955) of the genetic identities between the XJYN population (Ostrinia nubilalis Hübner, collected from Yining, Xinjiang Autonomous Region) and each of the other eight populations (O. furnacalis Guenée) was significantly smaller than that between the pairs of the eight populations (0.99386;t test,P<0.01). Although the population XJYN clearly deviates from the other eight populations in the dendrogram, the relationship of the two species of corn borer was very close. It is possible that the speciation of corn borer may have resulted from single-gene substitutions.     

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.     

Free amino acids of the haemolymph of the southwestern corn borer and the European corn borer in relation to their diapause

The titres of free amino acids present in the haemolymph of diapausing larvae of the southwestern corn borer, Diatraea grandiosella and the European corn borer, Ostrinia nubilalis, were examined. High titres of serine were found in the haemolymph of both species. Serine may serve as a storage form of compounds that are required for the synthesis of uric acid and other purines. The high titres of proline found in the haemolymph of O. nubilalis during the fall and winter may contribute to the freezing tolerance of this species. Alanine accumulated in the haemolymph of both species during the winter.     

Ancestral feeding and survival of offspring in European corn borer

Experiments were conducted to evaluate the influence of ancestral adult feeding by European corn borer, Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae), on larval survival in the field and neonate movement behavior in laboratory. Larval survival was higher when either the grandparental or parental generation had fed, but the feeding sites of the surviving larvae were not affected by ancestral feeding condition. This is the first evidence that grandparental feeding could influence larval survival in the field. Larval movement was observed in the laboratory. Silking speed of neonates was faster when either grandparents or parents had fed, while walking speed was faster only when parents had fed. No broad-sense genetic correlation was found between silking speed and walking speed. Broad-sense heritability among feeding histories were not significant for silking speed, but was significantly greater than zero for walking speed when grandparents fed and parents did not. These intergenerational effects could induce complex population dynamics in this species.     

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.     

Monitoring the Cry1Ab susceptibility of European corn borer in Germany

The European corn borer, Ostrinia nubilalis (Hübner), is one of the most important insect pests in corn, Zea mays L. Transgenic corn cultivars expressing Bacillus thuringiensis (Bt) toxin provide a promising crop protection strategy against European corn borer; however, management is needed to avoid resistance development of the target pest species. The aim of this work was to establish the baseline susceptibility of different European corn borer populations in Germany to be able to forecast a possible development of resistance at an early stage. To standardize test procedures for future resistance management, the efficiency of Cry1Ab toxins from different suppliers and different production was assessed. Furthermore, two different test methods, surface treatment and the incorporation method, were compared with regard to their practicability and efficiency. Neither method provided significant differences in the baseline susceptibility of populations from different German regions. Overall, the data suggested little differentiation among German populations in terms of their susceptibility to Bt toxin and their genetic background. Future monitoring could therefore use a single European corn borer population as a representative for southwestern Germany. However, toxins from different suppliers and different production batches produced a vast range of LC50 values. Changes because of different toxin batches may be mistaken as a change in baseline susceptibility or even as the start of a resistance development. Thus, it is important throughout insect resistance management that the same toxin batches will be available for baseline susceptibility bioassays and for future tests.     

Understanding successful resistance management: the European corn borer and Bt corn in the United States

The European corn borer, Ostrinia nubilalis Hübner (Lepidoptera: Crambidae) has been a major pest of corn and other crops in North America since its accidental introduction nearly a hundred years ago. Wide adoption of transgenic corn hybrids that express toxins from Bacillus thuringiensis, referred to as Bt corn, has suppressed corn borer populations and reduced the pest status of this insect in parts of the Corn Belt. Continued suppression of this pest, however, will depend on managing potential resistance to Bt corn, currently through the high-dose refuge (HDR) strategy. In this review, we describe what has been learned with regard to O. nubilalis resistance to Bt toxins either through laboratory selection experiments or isolation of resistance from field populations. We also describe the essential components of the HDR strategy as they relate to O. nubilalis biology and ecology. Additionally, recent developments in insect resistance management (IRM) specific to O. nubilalis that may affect the continued sustainability of this technology are considered.     

European corn borer injury effects on lignin, carbon and nitrogen in corn tissues

Plant herbivores often stimulate lignin deposition in injured plant tissue, but it is not known whether corn (Zea mays L.) reacts to European corn borer (ECB, Ostrinia nubilalis Hubner) injury in this manner. Bt (Bacillus thuringiensis) genetic modification is also reported to affect lignin in corn. This study evaluated the effects of ECB injury and the Bt gene on the chemical composition and decomposition of corn tissues. Eight near isolines (Bt and NBt) were grown in pots and half were infested with ECB. The experiment was repeated in 2 years. ECB injury increased the lignin concentration in corn leaves in one of 2 years and lowered the C:N ratio in injured stems. Lignin concentration in leaves was greater in Bt than NBt corn in 1 year and Bt stems had greater N concentration than NBt stems in 1 year of the 2 year study. ECB injury affected the composition of lignin-derived phenols, however ECB infested and non-infested stems lost the same amount of mass after 5 months in buried field litterbags. In conclusion ECB injury and the Bt gene had subtle effects on the chemical composition of corn tissue, which did not alter the short-term decomposition of corn residues.     

Salivary signals of European corn borer induce indirect defenses in tomato

Plants can recognize the insect elicitors and activate its defense mechanisms. European Corn Borer (ECB; Ostrinia nubilalis) saliva, produced from the labial salivary glands and released through the spinneret, is responsible for inducing direct defenses in host plants. Glucose oxidase (GOX) present in the ECB saliva induced direct defenses in tomato. By contrast, GOX activity in ECB saliva was insufficient to trigger defenses in maize, suggesting that host-specific salivary elicitors are responsible for inducing direct defenses in host plants. Our current study further examined whether ECB saliva can trigger indirect defenses in tomato. Relative expression levels of TERPENE SYNTHASE5 (TPS5) and HYDROPEROXIDE LYASE (HPL), marker for indirect defenses in host plants, were monitored. Quantitative real-time PCR analysis revealed that ECB saliva can induce the expression of TPS5 and HPL, suggesting that salivary signals can induce indirect defenses in addition to the direct defenses. Further experiments are required to identify different ECB elicitors that are responsible for inducing direct and indirect defenses in host plants.     

Population consequences of diapause in a model system: the European corn borer

The diapause biology of the European corn borer (ECB), Ostrinia nubilalis (Hübn.), is described based on natural and controlled environment studies of feral and lab-reared ECB's in North Carolina (NC). The diapause response is described as a function of photophase (h of light/day) as well as a function of larval age (instar) at onset of diapause-inducing conditions. A critical photophase of 14.4 h and a critical mean larval instar of 3.3 is found in the lab studies and supported by three years of insectary studies. Seven years of black light trapping of ECB moths in Goldsboro, NC, revealed the likelihood of up to four moth flights/year.Information about the diapause biology of this insect is used to explain both the number of flights and the relative magnitude of the final moth flights. On average, the majority of ECB lineages pass through three generations/year with early maturing ECB's producing a significant and predictable fourth generation. The timing and magnitude of the fourth flight can be partly explained on the basis of the critical photophase and the timing and age structure of previous ECB generations. In most years, the fourth flight is smaller than the third due to the majority of the fourth generation's predisposition towards diapause. However, in at least one case (1977), the fourth flight was unusually large and could be predicted by slight temporal shifts in the previous three flights resulting in the majority of the fourth generation larvae averting diapause. The value of the ECB-diapause interaction as a model system for the explanation and prediction of dynamic phenological events is discussed.

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Résumé La diapause d'O. nubilalis Hubn. est décrite d'après des études en conditions naturelles et programmées de souches sauvages ou élevées au laboratoire en Caroline du Nord. La diapause est décrite comme une fonction de la photophase (heures de jour/24 heures) et du stade larvaire au début des conditions inductrices de la diapause. Une photophase critique de 14,4 h et un stade critique larvaire de 3,3 ont été établis au laboratoire après 3 ans d'études en insectarium. 7 ans de piégeage à la lumière noire à Goldsboro, ont montré la vraisemblance de l'existence de 4 vols par an. Les données sur la diapause de cet insecte sont utilisées pour expliquer tant le nombre de vols que l'importance relative des derniers vols. En moyenne, la majorité des lignées ont 3 générations par an, O. nubilalis précoces produisant une quatrième génération conséquente et prédictible. La data et l'importance du 4ème vol peuvent être partiellement expliquées d'après la photophase critique, et la data et la structure en âge des générations précédentes. La plupart des années, le 4ème vol est moins important que le 3ème par suite de la prédisposition à la diapause de la majorité de la 4ème génération. Cependant, dans un cas au moins, en 1977, le 4ème était anormalement important et pouvait être prédit par de faibles changements temporels dans les 3 précédents vols, détournant de la diapause la majorité des chenilles de 4ème génération. La discussion porte sur la valeur du modèle fourni par la diapause de O. nubilalis pour expliquer et prédire la dynamique des évéments phénologiques.

     

Fungi associated with eggs and first-instar larvae of the European corn borer

Fungi isolated from field-collected egg masses of the European corn borer, Ostrinia nubilalis, were identified as Alternaria spp., A. porri, Fusarium spp., Fusarium oxysporum, Beauveria bassiana, Mucor spp., and an unidentified yeast. Most fungi were associated with predator injury to the egg mass. Bioassay of fungi on egg masses, however, showed that Alternaria spp. and A. porri reduced the hatch of both injured and uninjured egg masses, and Mucor sp. reduced the hatch only when the egg mass was injured.