Fine structure of endocrine hindgut cells of a lepidopteran,Ostrinia nubilalis (Hübn.)

Summary Cells of the hindgut of the European corn borer, Ostrinia nubilalis, have two functions, namely, ion transport and secretion of a hormone called proctodone. In this species, proctodone is an essential requisite to the prepupal molt in conjunction with brain hormone synthesis. Apical infoldings and their associated mitochondria form mitochondrial pumps for ion transport from the gut lumen to the hemocoel. The endocrine function of the cells is evidenced by rhythmical formation and discharge of inclusion bodies every 8 hours. These measure from 800 Å to 3 . in diameter and vary in composition from bodies with whorled, myeloid content to inclusions with densely granular matrix. During the discharge phase, granular material appears in the basal infoldings and accumulates in large quantities underneath the basal lamina and in the hemocoelic clefts adjacent to the active cells.This study constitutes publication No. 358 from the Oregon Regional Primate Research Center, supported in part by postdoctoral training fellowship 1-T1AM-5521-02 and Grant FR 00163 from the National Institutes of Health.     

Types of Antennal Sensilla of the Asian Corn Borer,Ostrinia furnacalis (Guenéee)

This study was carried out to investigate the types of sensilla and their distribution on the antennae of the Asian corn borer, Ostrinia furnacalis (Guenéee). O. furnacalis had six types of antennal sensilla: sensilla trichodea, sensilla chaetica, sensilla coeloconica, sensilla auricillica, sensilla styloconica and Böohms bristle. S. auricillica, single-walled multiporous sensilla, were presumed to be involved in chemical reception. S. coeloconica were double-walled, multiporous sensilla and could detect chemicals also. S. chaetica appeared to be mechano-receptors. S. styloconica were poreless and were thought to have thermo- and hygro-receptors. S. trichodea were single-walled chemosensilla and two subtypes were identified by transmission electron microscopy. The scape and the pedicel had Böohms bristles, which seem to function as mechanical stimulus receptors.     

Did the introduction of maize into Europe provide enemy‐free space to Ostrinia nubilalis? Parasitism differences between two sibling species of the genus Ostrinia

We examined whether maize offers enemy‐free space (EFS) to its pest Ostrinia nubilalis, and may thereby have contributed to its divergence from the sibling species, Ostrinia scapulalis, feeding mainly on mugwort, when introduced into Europe five centuries ago. We collected Ostrinia larvae on maize (70 populations, 8425 individuals) and mugwort (10 populations, 1184 individuals) and recorded parasitism using both traditional (counting emerging parasitoids) and molecular methods (detection by specific polymerase chain reaction). The main parasitoid was Macrocentrus cingulum (Braconidae). On mugwort, parasitism was twice that on maize, and parasitoid‐related mortality was 8 times higher. This suggests that maize affords substantial EFS to Ostrinia feeding on it. The lower Mortality:Infestation ratio in maize suggests that O. nubilalis’ immune response might be stronger than that of O. scapulalis. If so, adapting to maize and diverging from O. scapulalis would decrease the impact of parasitism on O. nubilalis at both ecological and evolutionary levels.     

Genetic isolation between two sympatric host-plant races of the European corn borer,Ostrinia nubilalis Hübner. I. Sex pheromone,moth emergence timing,and parasitism

Adaptation to different environments may be a powerful source of genetic differentiation between populations. The biological traits selected in each environment can pleiotropically induce assortative mating between individuals of these genetically differentiated populations. This situation may facilitate sympatric speciation. Successful host shifts in phytophagous insects provide some of the best evidence for the ecological speciation that occurs, or has occurred, in sympatry. The European corn borer, Ostrinia nubilalis (Lepidoptera: Crambidae), colonized maize after its introduction into Europe by humans about 500 years ago. In northern France, two sympatric host races feed on maize (Zea mays) and mugwort (Artemisia vulgaris), respectively. We investigated the factors involved in the genetic isolation of these two races at a field site near Paris, France. We identified two biological differences that might make a significant contribution to the genetic divergence between sympatric populations feeding on the two host plants. First, assortative mating may be due to differences in the moth emergence pattern between the two races: mugwort-race moths emerged on average 10 days earlier than maize-race moths. In addition, the males emerged earlier than females in both races. Hence, the likelihood of mating between maize-race males and mugwort-race females was higher than that of mating between mugwort-race males and maize-race females. Second, the females feeding on mugwort and maize produced sex pheromones with different E/Z isomeric ratios of delta-11-tetradecenyl acetate. This difference in mate recognition systems reinforces the potential for assortative mating in the two races. During the experiment, overwintering mortality was much lower on maize than on mugwort. This difference was due to a braconid parasitoid wasp, Macrocentrus cingulum, that killed more than 50% of the larvae overwintering on mugwort but did not infest larvae diapausing on maize. Hence, by colonizing maize, European corn borer populations probably escaped from numerous predators, competitors, and parasitoids, such as M. cingulum. This decrease in host-associated selection may have favored the colonization of this new host. Finally, throughout this experiment we observed selection at two allozyme loci (or at linked loci): Tpi and Mpi. The Tpi locus is tightly linked with the genes involved in the response of the male to the sex pheromone and in developmental timing. The location of these traits on the Z chromosome may play a role in shortening the time required for the evolution of premating barriers.     

Tissue Distribution and Purification of Prophenoloxidase in Larvae of Asian Corn Borer, Ostrinia furnacalis Guenée (Lepidoptera: Pyralidae)

Phenoloxidase (PO) plays an important role in the de-fense response to the foreign invaders, e.g. pathogens andparasites, in the formation of melanin as well as in cuticlesclerotization in insects [1]. PO is synthesized as azymogen, prophenoloxidase (PPO)…     

Distribution and variations of three 1,4-benzoxazin-3-ones in maize induced by the Asian corn borer, Ostrinia furnacalis (Guenée)

Contents of three 1,4-benzoxazin-3-ones in tissue samples from different parts (young leaf, second leaf, old leaf, stem and root) of young maize plants of 4-leaves stage, fed by the third instar larvae of the Asian corn borer, Ostrinia furnacalis (Guenée), were analyzed by high-performance liquid chromatography-mass spectroscopy (HPLC-MS). Samples were taken immediately (set A) or 48 h (set B) after larvae had fed on the second leaf for 48 h. The three 1,4-benzoxazin-3-ones investigated in our experiments were 2,4-dihydroxy-7-methoxy-1,4(2H)-benzoxazin-3-one (DIMBOA), 2,4-dihydroxy-1,4(2H)-benzoxazin-3-one (DIBOA) and 2-hydroxy-7-methoxy-1,4(2H)-benzoxazin-3-one (HMBOA). In samples of set A, the levels of DIMBOA and HMBOA were significantly lifted in the old leaf (L3) and young leaf (L1), respectively, while amounts of these two chemicals in other plant parts were not significantly different between larvae-fed plants and intact plants. Concentrations of DIBOA in each plant part remained unchanged. In samples of set B, no concentration differences for any of these three 1,4-benzoxazin-3-ones between larvae-fed plants and controls were observed in any plant part. The feeding of the Asian corn borer seems to have limited effects on induction of these three 1,4-benzoxazin-3-ones in young maize plants of the variety investigated.     

Suitability of three Ostrinia species as hosts for Macrocentrus cingulum： A comparison of their encapsulation abilities

Two cornborer species, Ostrinia furnacalis （Lepidoptera： Crambidae） and O. nubilalis, are major corn pests in Asia and Europe, respectively. In both continents, the larval endoparasitoid Macrocentrus cingulum （Hymenoptera： Braconidae） develops on another, closely related stemborer, O. scapulalis, which feeds on mugwort and other dicotyledons. M. cingulum also emerges from O. furnacalis in Asia and （9. nubilalis in North America, but not from O. nubilalis in Europe. We assessed the ability of three populations of each of the three Ostrinia species to encapsulate foreign bodies of a size similar to that of a M. cingulum egg. We conclude that variations in encapsulation ability alone cannot account for the differences observed in the field between parasite emergence rates in these different host species and geographic areas.     

PBAN对亚洲玉米螟(Ostrinia furnacalis)性外激素生物合成路线的控制(英)

去头处理致使亚洲玉米螟性外激素的含量显著地下降到很低的水平。注射雄性或雌性玉米螟头部提取物或合成PBAN(激活外激素生物合成神经肽)可使外激素的含量得以恢复。因此可知玉米螟外激素的产生系受一种类PBAN因子控制。玉米螟性外激素生物合成路线由棕榈酸的生物合成开始,然后经14位脱饱和化,碳链缩短,还原和乙酰化形成外激素顺和反12—十四碳烯乙酸酯。为了阐明受PBAN控制的生物合成步骤,研究了不同的标记前体掺入外激素及其中间体的情况。根据结果推论,PBAN主要通过调节由乙酸酯到棕榈酸的生物合成步骤来控制外激素生物合成。     

Der Einfluss des Wirtes in der Massenzucht auf die Qualität des EiparasitenTrichogramma evanescens bei der Bekämpfung des Maiszünslers,Ostrinia nubilalis

Zusammenfassung In vier Feldversuchen wurde die Wirksamkeit eines an den Maiszünsler besonders angepassten Stammes vonTrichogramma evanescens Westw. bei der Bek?mpfung des MaiszünslersOstrinia nubilalis Hübner geprüft. Die Parasiten wurden entweder auf Eiern der GetreidemotteSitotroga cerealella (Oliv.) oder auf Eiern der MehlmotteEphestia kuehniella Zell. gezüchtet. Die Freilassung erfolgte in den Versuchsparzellen durch Anbringen von Eikarten mit jeweils etwa 1000 parasitierten Wirtseiern an Bl?ttern von Maispflanzen in Abst?nden von 14 Metern. VierTrichogramma-Freilassungen in w?chentlichen Abst?nden mit jeweils 45000 Parasiten pro Hektar ergaben hohe Parasitierungsraten bei denO. nubilalis-Eiern und starke Verminderungen des Larvenbefalls. In den Parzellen, in denen aufS. cerealella-Eiern gezüchtete Parasiten freigelassen wurden, lagen die Parasitierungsraten zwischen 81,7 und 82,3% im Vergleich zu 63,0 und 84,3% bei den aufE. kuehniella gezüchteten Trichogrammen. Die Verminderung des bei der Ernte festgestelltenO. nubilalis-Larvenbefalls im Vergleich zur unbehandelten Kontrolle lag bei den aufS. cerealella gezüchteten Parasiten zwischen 75,5 und 94,8% verglichen mit 77,6 und 89,9% bei den aufE. kuehniella gezüchteten Tieren. Es gab somit keine signifikanten Unterschiede zwischen den beiden getestetenTrichogramma-St?mmen.

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Summary The effectiveness of a strain ofTrichogramma evanescens Westw. especially adapted to the European corn borerOstrinia nubilalis Hübner was examined in 4 field experiments. The parasites were reared on either the Angoumois grain mothSitotroga cerealella (Oliv.) or the Mediterranean flour mothEphestia kuehniella Zell. and were released in separate experimental plots by attaching egg cards, each containing about 1000 parasitized host eggs, to corn leaves at distances of 14 meters in the field. FourTrichogramma treatments at 1 week intervals using 45000 parasites per hectar per treatment resulted in high parasitization rates ofO. nubilalis-eggs and large reductions in larval damage. In experimental plots whereT. evanescens, reared onS. cerealella-eggs, were released, the parasitization rates ofO. nubilalis-eggs varied between 81,7 and 82,3%, compared with 63,0 and 84,3% forT. evanescens reared onE. kuehniella. The reduction inO. nubilalis larval infestation at the harvest in the treated plots, compared to untreated controls varied between 75,5 and 94,8% forT. evanescens that was reared onS. cerealella, compared with 77,6 and 89,8% forT. evanescens that was reared onE. kuehniella. There were no significant differences in efficiency between the two tested methods of rearingTrichogramma.

     

Fatty acid and sex pheromone changes and the role of glandular lipids in the Z-strain of the European corn borer, Ostrinia nubilalis (Hübner)

Lipids in the sex pheromone gland of females of the Z-strain of Ostrinia nubilalis were analyzed for fatty acyl pheromone analogs (FAPAs) and other potential biosynthetic intermediates. More than 80% of the FAPAs were found in the triacylglycerols (TGs), with smaller amounts found in the phosphatidyl cholines, ethanolamines, and serines. Analysis of the TGs by lipase revealed that the two FAPAs were distributed fairly evenly among all three stereospecific positions. Comparison of changes in titers of key glandular fatty acids with those of pheromone components, with respect to photoperiodic time and age of females, showed that both FAPA and pheromone titers exhibited a cyclical pattern with peaks in the scotophase and valleys in the photophase. However, whereas pheromone titer tended to peak in the first half of the scotophase, FAPA titer peaked at the end of the scotophase. Significantly, the titer of the FAPA of the minor component, (E)-11-tetradecenyl acetate (3% of pheromone), was always much greater than the titer of the FAPA of the major component, (Z)-11-tetradecenyl acetate (97%), of the pheromone. Titer of myristate, an intermediate in pheromone biosynthesis, was also higher during the scotophase than the photophase. However, myristate titer showed a pronounced dip in the middle of the scotophase. These data suggest two roles for glandular lipids in sex pheromone biosynthesis in O. nubilalis. Firstly, they remove excess FAPA of the minor component so the fatty acid reductase system is not presented with a high ratio of this isomer (which would otherwise result from the reductase's own selectivity), which could cause changes in the final pheromone ratio. Secondly, hydrolysis of the large amounts of stored saturated fatty acids from the TGs may provide substrate for pheromone biosynthesis.     

Genetic structure and gene flow in French populations of two Ostrinia taxa: host races or sibling species?

Most models of ecological speciation concern phytophagous insects in which speciation is thought to be driven by host shifts and subsequent adaptations of populations. Despite the ever-increasing number of studies, the current evolutionary status of most models remains incompletely resolved, as estimates of gene flow between taxa remain extremely rare. We studied the population genetics of two taxa of the Ostrinia genus--one feeding mainly on maize and the other on mugwort and hop--occurring in sympatry throughout France. The actual level of divergence of these taxa was unknown because the genetic structure of populations had been investigated over a limited geographical area and the magnitude of gene flow between populations had not been estimated. We used 11 microsatellite markers to investigate the genetic structure of populations throughout France and the extent of gene flow between the two Ostrinia taxa at several sites at which they are sympatric. We observed clear genetic differentiation between most populations collected on the typical respective hosts of each taxon. However, populations displaying intermediate allelic frequencies were found on hop plants in southern France. Individual assignments revealed that this result could be accounted for by the presence of both taxa on the same host. Gene flow, estimated by determining the proportion of hybrids detected, was low: probably<1% per generation, regardless of site. This indicates that the two Ostrinia taxa have reached a high level of genetic divergence and should be considered sibling species rather than host races.     

Host-plant diversity of the European corn borer Ostrinia nubilalis: what value for sustainable transgenic insecticidal Bt maize?

The strategies proposed for delaying the development of resistance to the Bacillus thuringiensis toxins produced by transgenic maize require high levels of gene flow between individuals feeding on transgenic and refuge plants. The European corn borer Ostrinia nubilalis (Hübner) may be found on several host plants, which may act as natural refuges. The genetic variability of samples collected on sagebrush (Artemisia sp.), hop (Humulus lupulus L.) and maize (Zea mays L.) was studied by comparing the allozyme frequencies for six polymorphic loci. We found a high level of gene flow within and between samples collected on the same host plant. The level of gene flow between the sagebrush and hop insect samples appeared to be sufficiently high for these populations to be considered a single genetic panmictic unit. Conversely, the samples collected on maize were genetically different from those collected on sagebrush and hop. Three of the six loci considered displayed greater between-host-plant than within-host-plant differentiation in comparisons of the group of samples collected on sagebrush or hop with the group of samples collected on maize. This indicates that either there is genetic isolation of the insects feeding on maize or that there is host-plant divergent selection at these three loci or at linked loci. These results have important implications for the potential sustainability of transgenic insecticidal maize.     

Two Different Bacillus thuringiensis Delta-Endotoxin Receptors in the Midgut Brush Border Membrane of the European Corn Borer, Ostrinia nubilalis (Hübner) (Lepidoptera: Pyralidae)

Binding of three Bacillus thuringiensis insecticidal crystal proteins (ICPs) to the midgut epithelium of Ostrinia nubilalis larvae was characterized by performing binding experiments with both isolated brush border membrane vesicles and gut tissue sections. Our results demonstrate that two independent ICP receptors are present in the brush border of O. nubilalis gut epithelium. From competition binding experiments performed with I-labeled and native ICPs it was concluded that CryIA(b) and CryIA(c) are recognized by the same receptor. An 11-fold-higher binding affinity of CryIA(b) for this receptor correlated with a 10-fold-higher toxicity of this ICP compared with CryIA(c). The CryIB toxin did not compete for the binding site of CryIA(b) and CryIA(c). Immunological detection of ingested B. thuringiensis ICPs on gut sections of O. nubilalis larvae revealed binding only along the epithelial brush border membrane. CryID and CryIE, two ICPs that are not toxic to O. nubilalis, were not bound to the apical microvilli of gut epithelial cells. In vitro binding experiments performed with native and biotinylated ICPs on tissue sections confirmed the correlation between ICP binding and toxicity. Moreover, by performing heterologous competition experiments with biotinylated and native ICPs, it was confirmed that the CryIB receptor is different from the receptor for CryIA(b) and CryIA(c). Retention of activated crystal proteins by the peritrophic membrane was not correlated with toxicity. Furthermore, it was demonstrated that CryIA(b), CryIA(c), and CryIB toxins interact in vitro with the epithelial microvilli of Malpighian tubules. In addition, CryIA(c) toxin also adheres to the basement membrane of the midgut epithelium.     

Development and characterization of a continuous cell line,AFKM-On-H,from hemocytes of the European corn borer <Emphasis Type="BoldItalic">Ostrinia nubilalis</Emphasis> (Hübner) (Lepidoptera,Pyralidae)

The corn borer, Ostrinia nubilalis, is a very important pest in different countries, and the in vitro system of the insect could be a useful tool for isolation and characterization of the pathogens and physiological responses of the insect. In this context, a cell line was derived from the hemocytes of the European corn borer and was named AFKM-On-H for, respectively, O. nubilalis, Armand Frappier, King Mongkut Institutes, and Hemocytes. This cell line was initiated and maintained in Ex-Cell 400 medium supplemented with 10% heat-inactivated fetal bovine serum. The cells, mostly spherical in shape, not firmly attached to the plastic culture flasks, were passaged up to 200 times by repeated gentle pipetting of the cells. The doubling times at the 80th and 125th passages at 28°C and at the 122th and 169th passages at 25°C were 40, 29, 35, and 34 h, respectively. The AFKM-On-H cell line was further characterized by the morphology, karyotype, random amplified polymorphic DNA analysis, and isozyme profiles. Susceptibility of the cell line to cytoplasmic polyhedrosis viruses (CPV) Euxoa scandens (EsCPV), Dendrolimus punctatus (DpCPV), and Choristoneura fumiferana (CfCPV); nuclear polyhedrosis viruses [Autographa californica (AcMNPV) wild type and recombinant, Antherea yammamai (AnyaNPV)]; and Chilo iridescent virus was demonstrated. Relative sensitivities of the cell line to Bacillus thuringiensis and Metarhizium anisopliae toxins and effects of the molting hormone 20-hydroxyecdysone on this new hemocyte cell line were characterized.     

PERMANENT GENETIC RESOURCES: Isolation and characterization of microsatellite loci from the European corn borer, Ostrinia nubilalis (Hübner) (Insecta: Lepidoptera: Crambidae)

Few useful microsatellites are available for population studies of the European corn borer, Ostrinia nubilalis (Hübner). An enrichment strategy was used to develop microsatellite markers for O. nubilalis, and over 500 positive clones were isolated. Seventy-five contained unique microsatellites, 10 of which were polymorphic with discernable polymerase chain reaction products. The 10 loci were surveyed for variability in 72 wild individuals from central Iowa. Five loci showed no deviation from Hardy-Weinberg proportions, and all were successfully cross-amplified in the related Asian corn borer, Ostrinia furnacalis. These loci represent a significant addition to microsatellites appropriate for population studies of O. nubilalis.