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Ostrinia nubilalis, a lepidopteran moth, also known as the European corn borer, has a major impact on the production of economically important crops in the United States and Europe. The female moth invites the male moth for mating through the release of pheromones, a volatile chemical signal. Pheromone binding proteins (PBPs) present in the male moth antennae are believed to pick up the pheromones, transport them across the aqueous sensillum lymph, and deliver them to the olfactory receptor neurons. Here we report for the first time the cloning, expression, refolding, purification, and structural characterization of Ostrinia nubilalis PBP3 (OnubPBP3). The recombinant protein showed nanomolar affinity to each isomer of the Ostrinia pheromones, E- and Z-11-tetradecenyl acetate. In a pH titration study by nuclear magnetic resonance, the protein exhibited an acid-induced unfolding at pH below 5.5. The molecular dynamics simulation study demonstrated ligand-induced conformational changes in the protein with both E- and Z-isomers of the Ostrinia pheromone. The simulation studies showed that while protein flexibility decreases upon binding to E-pheromone, it increases when bound to Z-pheromone. This finding suggests that the OnubPBP3 complex with E-pheromone is more stable than with Z-pheromone.

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., We compared egg production within and between corn borer lines derived from populations that differ in pheromone similarity. Our results suggest that elevated temperature increases investment among populations. At elevated temperature (5°C above the pupal environment), between‐line pairs produced the same amount of egg clusters as within‐line pairs unlike at ambient temperatures (matching the pupal environment).

European corn borer (Ostrinia nubilalis) is an important maize pest which is treated mainly by synthetic pesticides with many negative effects for ecosystem and human health. Soil borne entomopathogenic fungi of species belonging to Beauveria genus may provide an alternative strategy for biological control of this pest. In this article, we report results of virulence testing of 46 soil isolates of Beauveria spp. against O. nubilalis in relation to isolates origin and haplotype. All strains were collected from soil using Galleria mellonella bait method. Conidia suspension with concentration 10⁷ conidia in ml was used for virulence test against 4th instar larvae of O. nubilalis at temperature 25°C. After 14 days, mortality of larvae was in the range 34%–96%. Median lethal time LT50 was estimated to 5.5–21.3 days. Mortality was not in relation to habitat or any other environmental parameter of sites where isolates came from. Differences among species were insignificant. Isolates were divided into 8 genetic lineages and 14 haplotypes using sequencing of ITS and Bloc nuclear regions. None of the lineages showed higher mortality than others. Despite more or less virulent isolates being found in each lineage, the three most virulent isolates belonged to the same lineage Beauveria bassiana A3. The four most virulent isolates were compared to commercial isolate GHA to assess medium lethal concentration. LC50 were estimated in range 7.5 × 10⁴ to 9.5 × 10⁵. Three of the four isolates showed equal or greater efficiency than strain GHA.

The European corn borer (ECB; Ostrinia nubilalis) is an economically damaging insect pest of maize (Zea mays L.), an important cereal crop widely grown globally. Among inbred lines, the maize genotype Mp708 has shown resistance to diverse herbivorous insects, although several aspects of the defense mechanisms of Mp708 plants are yet to be explored. Here, the changes in root physiology arising from short-term feeding by ECB on the shoot tissues of Mp708 plants was evaluated directly using transcriptomics, and indirectly by monitoring changes in growth of western corn rootworm (WCR; Diabrotica virgifera virgifera) larvae. Mp708 defense responses negatively impacted both ECB and WCR larval weights, providing evidence for changes in root physiology in response to ECB feeding on shoot tissues. There was a significant downregulation of genes in the root tissues following short-term ECB feeding, including genes needed for direct defense (e.g., proteinase inhibitors and chitinases). Our transcriptomic analysis also revealed specific regulation of the genes involved in hormonal and metabolite pathways in the roots of Mp708 plants subjected to ECB herbivory. These data provide support for the long-distance signaling-mediated defense in Mp708 plants and suggest that altered metabolite profiles of roots in response to ECB feeding of shoots likely negatively impacted WCR growth.

The microsporidium Nosema pyrausta is an important mortality factor of the European corn borer, Ostrinia nubilalis. The present study was aimed at N. pyrausta virulence testing to the beet webworm (BW), Loxostege sticticalis. This agricultural pest, L. sticticalis, was highly vulnerable to N. pyrausta. The parasite’s spores were located in salivary glands, adipose tissue, and Malpighian tubules of the infected specimens. Infection was transmitted transovarially through at least 3 laboratory generations, in which BW fitness indices were lower than in the control, and moth emergence and fertility decreased prominently. Transovarial infection was most detrimental to female egg-laying ability, resulting in zero fertility in F3. When propagated in BW, the microsporidium tended to increase its virulence to L. sticticalis, as compared to the Ostrinia isolates. The parasite’s ability to infect this host at low dosages and transmit vertically should guarantee its effective establishment and spread within BW populations. In conclusion, N. pyrausta is a promising agent against BW, which is a notorious polyphagous pest in Eurasia.

Long-term observations have revealed the factors responsible for periodic rises in abundance of the European corn borer Ostrinia nubilalis Hb. in the Central Chernozem Region of Russia. These factors are sufficient precipitation during several consecutive years and the current decline in agrotechnical soil treatment. The host specialization, biology, and phenology of the corn borer in the region have been clarified. Corn is damaged most intensely among the cultivated plants, and the yellow foxtail Setaria glauca, among the weeds. The effects of precipitation on seasonal development and harmfulness of the corn borer have been demonstrated. Low precipitation during the pest pupation period leads to mass pupal mortality, a shift of the seasonal development onto later dates, and a lower proportion of damaged stems in the crops. Insufficient precipitation during the larval feeding period enhances the effect of pest damage on the plant productivity and results in heavier yield loss. During the corn borer outbreaks the yield loss reached 15% in corn and 11.4% in millet. Depending on the weather conditions, the loss of millet yield due to this pest may be as great as 39% or as low as 3%.

The European corn borer, Ostrinia nubilalis (Lepidoptera: Crambidae), is a perennial insect pest of cultivated maize that was inadvertently introduced into North America in the early 1900s, but population densities have decreased since the widespread adoption of transgenic hybrids that express Bacillus thuringiensis (Bt) toxins. The native American lotus borer, Ostrinia penitalis (Lepidoptera: Crambidae), is among the most ancestral species described in the genus Ostrinia, and has a geographic range that coincides with that of O. nubilalis across major maize growing regions of North America. Due to the recent decrease in O. nubilalis populations, O. penitalis has become more pronounced in light trap samples intended to monitor O. nubilalis. A molecular tool based on variation in restriction endonuclease digestion pattern of a polymerase chain reaction amplified fragment of the mitochondrial cytochrome c oxidase subunit I (coxI) gene was developed and validated to differentiate these two species. This method was applied to light trap samples over a 2-yr period and achieved accurate quantification of species, and shows that O. penitalis can be prevalent in O. nubilalis first flight sampling. These methods are useful for contemporary O. nubilalis field research in North America.

International audience; The olfactory choices expressed by naive neonate European corn borer (ECB) larvae were tested by recording their locomotor activities in response to odours coming from plants of the genus Chenopodium (producing phytoecdysteroids or not). ECB larvae were found to be repelled from phytoecdysteroid-positive species, except C. album and C. polyspermum. On the contrary, they were mildly attracted or mildly repelled by phytoecdysteroid-negative species, except C. botrys (which emits a rancid odour). These observations indicate that neonate ECB larvae clearly differentiate plant odours and suggest that well-defended plants may emit repellent odours.

The gypsy moth, Lymantria dispar, is a notorious forest defoliator, and various pathogens are known to act as natural regulators of its population density. As a widespread herbivore with a broad range of inhabited areas and host plants, it is potentially exposed to parasitic microorganisms from other insect hosts. In the present paper, we determined the susceptibility of gypsy moth larvae to the microsporidium Nosema pyrausta from the European corn borer, Ostrinia nubilalis. Gypsy moth samples from two localities of Western Siberia were used. N. pyrausta developed infections in the salivary gland and adipose tissue of gypsy moth prepupae and pupae, forming spore masses after 30 days of alimentary exposure to the second instar larvae. Among the experimental groups, the infection levels ranged from 0 to 9.5%. Effects of a covert baculovirus infection, phenylthiourea pretreatment and feeding insects on an artificial diet versus natural foliage were not significant in terms of microsporidia prevalence levels. Thus, L. dispar showed a low level of susceptibility to a non-specific microsporidium. It can be referred to as a resistant model host and not an appropriate substitute host for laboratory propagation of the microsporidium.

Background The European corn borer (ECB), Ostrinia nubilalis (Hübner, 1796) (Lepidoptera: Crambidae), is the major pest of maize (Zea mays Linnaeus, 1753) in Serbia. One potential method for managing this pest is the augmentative release of naturally occuring egg parasitoids of the genus Trichogramma. The first step in this process is accurately identifying the naturally occuring species and estimating their natural distribution and abundance. Molecular identification, based on differences in DNA sequences, has commonly been employed for the identification of Trichogramma species. A simple, quick, and accurate molecular assay is urgently required for the identification of two common Trichogramma species, associated with ECB in Serbia: T. brassicae Bezdenko, 1968 and Trichogramma evanescens Westwood, 1833. Such an assay will facilitate an expansive survey of resident populations of Trichogramma associated with ECB across agricultural growing regions of Vojvodina province. Results A species-specific multiplex PCR assay for the 2 species was developed and validated that assay using a sample of 79 parasitoid wasps reared from ECB egg masses collected from sample sites across Vojvodina province. Trichogramma brassicae was confirmed as the dominant egg parasitoid of ECB in this region, accounting for 77 of the 79 wasps (97.47%). The remaining 2 were confirmed as T. evanescens. Trichogramma brassicae was detected at all 12 sample sites, while T. evanescens was detected at only 2 plots, Mokrin and Nakovo. Conclusions The species-specific multiplex PCR assay presented herein can provide the basis of a quick, cheap, and reliable means for identifying the species of Trichogramma that parasitize ECB egg masses in Serbia. Two currently documented species, T. brassicae and T. evanescens, are readily diagnosed by the size of the PCR product they produce in the assay. Any additional species are expected to not produce a band of a diagnostic size. Such species would subsequently be identified by sequencing, which may also allow them to be promptly incorporated into a revised assay.

Maize with enhanced β-carotene production was engineered to counteract pervasive vitamin A deficiency in developing countries. Second-generation biofortified crops are being developed with additional traits that confer pest resistance. These include crops that can produce Bacillus thuringiensis Berliner (Bt) insecticidal proteins. Currently, it is unknown whether β-carotene can confer fitness benefits through to insect pests, specifically through altering Ostrinia nubilalis foraging behaviour or development in the presence of Bt insecticidal toxin. Therefore the effects of dietary β-carotene plus Bt insecticidal protein on feeding behaviour, mortality, and physiology in early and late instars of O. nubilalis larvae were investigated. The results of two-choice experiments showed that irrespective of β-carotene presence, at day five 68%-90% of neonates and 69%-77% of fifth-instar larvae avoided diets with Cry1A protein. Over 65% of neonate larvae preferred to feed on diets with β-carotene alone compared to 39% of fifth-instar larvae. Higher mortality (65%-97%) in neonates fed diets supplemented with β-carotene alone and in combination with Bt protein was found, whereas

International audience; The current study extends the faunistic survey ofTrichogrammaspecies parasitizing ECB to all agricultural growing regions in Serbia. Specimens ofTrichogrammawere reared from parasitized egg masses of ECB collected from field-grown corn and pepper crops. The number of egg masses parasitized varied by location. Using sequences of mitochondrial cytochrome oxidase I (COI) gene, we examined intra and interspecific variation inTrichogrammaparasitizing the ECB egg masses. Seventy specimens were successfully sequenced of which, 65 were identified asT. brassicae. The remaining five were identified asT. evanescens, and this second species was only collected from pepper.

Slow and consistent nutrient release by organic fertilizers can improve plant nutrient balance and defenses, leading to herbivore avoidance of organically managed crops in favor of conventional crops with weaker defenses. We propose that this relative attraction to conventional plants, coupled with the use of genetically modified, insecticidal crops (Bt), has created an unintentional attract-and-kill system. We sought to determine whether Bt and non-Bt corn Zea mays L. plants grown in soil collected from five paired organic and conventional fields differed in attractiveness to European corn borer [Ostrinia nubilalis (Hübner)] moths, by conducting ovipositional choice and flight tunnel assays. We then examined the mechanisms driving the observed differences in attraction by comparing soil nutrient profiles, soil microbial activity, plant nutrition, and plant volatile profiles. Finally, we assessed whether European corn borer abundance near corn fields differed based on soil management. European corn borer preferred plants grown in conventional soil but did not discriminate between Bt and non-Bt corn. Organic management and more alkaline soil were associated with an increased soil magnesium:potassium ratio, which increased plant magnesium, and were linked to reduced European corn borer oviposition. There was an inconsistent trend for higher European corn borer moth activity near conventional fields. Our results extend the mineral balance hypothesis describing conventional plant preference by showing that it can also improve attraction to plants with genetically inserted toxins. Unintentional attract (to conventional) and (Bt) kill is a plausible scenario for pest declines in response to Bt corn adoption, but this effect may be obscured by variation in other management practices and landscape characteristics.

RNA interference (RNAi) is a revolutionary technique for silencing gene expression, but the success of this technique is dependent upon the stability of double-stranded RNA (dsRNA) molecules. In many insects, especially lepidopteran species, RNAi efficiency is limited by high instability of dsRNA in the gut and/or hemolymph, preventing the development of RNAi-based strategies for many serious pests. Previous attempts to perform RNAi on Ostrinia nubilalis (ECB, Lepidoptera: Crambidae) indicate low RNAi efficiency with both dsRNA injection and feeding. To investigate the contribution of dsRNA instability to low RNAi efficiency in ECB, a serious of ex vivo incubation experiments were performed where dsRNA integrity was assessed following incubation in larval gut continents and hemolymph using gel electrophoresis or RT-qPCR. DsRNA was less stable in the gut contents from ECB than in gut contents from Diabrotica virgifera virgifera, a coleopteran exhibiting high RNAi efficiency. Furthermore, characterization of dsRNA stability in ECB gut contents and hemolymph revealed that dsRNA was rapidly degraded under physiologically relevant conditions as a result of enzymatic activity that was neither size- nor sequence-dependent. These findings suggest that instability of dsRNA in ECB tissues is a contributing factor to the poor efficiency of RNAi in this pest. This work advances our understanding of mechanisms impacting RNAi efficiency in ECB and related lepidopteran insects for which novel pest management strategies are needed, and may facilitate the development of strategies for enhancing dsRNA stability in ECB tissues.

Bracon brevicornis is an ectoparasitoid of a wide range of larval-stage Lepidopterans, including several pests of important crops, such as the corn borer, Ostrinia nubilalis. It is also one of the earliest documented cases of complementary sex determination in Hymenoptera. Here, we present the linked-read-based genome of B. brevicornis, complete with an ab initio-derived annotation and protein comparisons with fellow braconids, Fopius arisanus and Diachasma alloeum. We demonstrate the potential of linked-read assemblies in exploring regions of heterozygosity and search for structural and homology-derived evidence of the complementary sex determiner gene (csd).

The genus Ostrinia (Lepidoptera; Crambidae) consists of 21 species worldwide, 9 of which inhabit Japan. These closely related species, which utilize different host plants, provide an excellent model for studies on the evolution of sex pheromone communication systems as well as speciation. In this chapter, we review the progress of research on the sex pheromone communication system of Ostrinia inhabiting Japan. We focus on the molecular bases for the biosynthesis of sex pheromone components, differential production of which is responsible for the divergence of sex pheromones.

The European corn borer, Ostrinia nubilalis Hbn., is a pest Lepidopteran species whose larvae overwinter by entering diapause, gradually becoming cold-hardy. To investigate metabolic changes during cold hardening, activities of four metabolic enzymes – citrate synthase (CS), lactate dehydrogenase (LDH), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were measured in whole-body homogenates of pupae, non-diapausing and diapausing larvae acclimated to 5 °C, −3 °C and −16 °C. The highest CS activity was detected in non-diapausing larvae, reflecting active development, while the highest in vitro LDH activity was recorded in diapausing larvae at temperatures close to 0 °C, evidencing a metabolic switch towards anaerobic metabolism. However, in-gel LDH activity showed that production of pyruvate from lactate is triggered by sub-zero temperatures. The activities of both aminotransferases were highest in non-diapausing larvae. Our findings suggest that during diapause and cold hardening the aminotransferases catalyse production of L-alanine, an important cryoprotectant, and L-aspartate, which is closely tied to both transamination reactions and Krebs cycle. The results of this study indicate that, during diapause, the activity of metabolic enzymes is synchronized with exogenous factors, such as temperatures close to 0 °C. These findings support the notion that diapause is metabolically plastic and vibrant, rather than simply a passive, resting state.

Transgenic maize, Zea mays L., modified to express insecticidal proteins from the bacterium Bacillus thuringiensis Berliner, was introduced in 1996 to control Ostrinia nubilalis Hübner (Lepidoptera: Crambidae), a key maize pest in North America. The high-dose/refuge concept, developed to delay or prevent resistance evolution to this technology, has been exemplified by O. nubilalis as no cases of practical resistance were identified in >20 years. This study documents the first case of practical resistance to Cry1F Bt maize by O. nubilalis in North America. Four collections of O. nubilalis were made from Cry1F maize in Nova Scotia, Canada with unexpected injury (UXI) ranging from 30–70%. Greater survival of UXI collections was observed when larvae were exposed to the highest concentration of 200 ng Cry1F cm−2 in diet-overlay bioassays compared to susceptible laboratory colonies. Larvae also fed and survived on Cry1F leaf tissue in 7 d bioassays. A collection from non-Bt maize, 120 km west of the UXI region, also survived 200 ng Cry1F cm−2, but was susceptible to Cry1F leaf tissue. Detection of Cry1F-resistant O. nubilalis in what might be considered an insignificant maize-growing region indicates that a number of preventable causal factors may have been related to inadequate stewardship of Bt maize technology.

Analysis of the prevalence rates of the endosymbiotic bacterium of the Wolbachia genus in the populations of corn borers of the Ostrinia genus was carried out. Samples of genomic DNA were extracted from field-collected larvae. The PCR analysis for the presence of Wolbachia infection was performed independently using three diagnostic loci: 16SrRNA, gatB, and fbpA. A positive signal for Wolbachia was obtained for 13.5% of the Ostrinia nubilalis larvae (N = 141) and 31.9% of the Ostrinia scapulalis larvae (N = 138). The infection rate ranged from 2.9% (N = 34) to 65.8% (N = 38) in different geographical populations. At the same time, a significantly higher infection rate of O. scapulalis than O. nubilalis was observed in three out of four habitats. It was also found that the Wolbachia infection was more common for O. scapulalis individuals feeding on hemp than for those collected from mugwort.

A comparison of the small subunit rRNA gene sequences of the microsporidian isolates from corn borers of the Ostrinia genus from Russia and Belarus indicates that they belong to the Nosema pyrausta species. In order to analyze genetic polymorphism, the nucleotide sequences of the molecular clones of the rRNA intergenic spacer (IGS) of the N. pyrausta isolates from Ostrinia nubilalis and Ostrinia scapulalis (Krasnodar territory), as well as from O. nubilalis (Gomel region, Belarus), are determined. Based on the information obtained and available in Genbank, a high level of divergence of the IGS locus between the isolates of different species and populations of microsporidia, as well as between the molecular clones within individual isolates of parasites, is shown.

Transgenic maize hybrids that express insecticidal Bacillus thuringiensis (Bt) crystalline (Cry) protein toxins effectively protect against the European corn borer, Ostrinia nubilalis, a devastating maize pest. Field monitoring and laboratory selections have detected varying levels of O. nubilalis resistance to Cry1Ab toxin. MicroRNAs (miRNAs) are short noncoding RNAs that are involved in post-transcriptional gene regulation. Their potential roles in the evolution of Bt resistance, however, remain largely unknown. Sequencing of small RNA libraries from the midgut of Cry1Ab-susceptible and resistant O. nubilalis larvae resulted in the discovery of 277 miRNAs, including 248 conserved and 29 novel. Comparative analyses of miRNA expression profiles between the laboratory strains predicted 26 and nine significantly up- and down-regulated transcripts, respectively, in the midgut of Cry1Ab resistant larvae. Amongst 15 differentially regulated miRNAs examined by quantitative real-time PCR, nine (60%) were validated as cosegregating with Cry1Ab resistance in a backcross progeny. Differentially expressed miRNAs were predicted to affect transcripts involved in cell membrane components with functions in metabolism and binding, and the putative Bt-resistance genes aminopeptidase N and cadherin. These results lay the foundation for future investigation of the potential role of miRNAs in the evolution of Bt resistance.

Background Understanding interactions between biocontrol agents and their pest hosts under climate change should assist implementation of biocontrol strategies, by identifying appropriate biocontrol agents for release or determining the optimal timing of releases. Species distribution models (SDMs) were applied to evaluate the distributions of Trichogramma ostriniae and its native host, Ostrinia furnacalis, in southeastern Asia, and a non-native host, Ostrinia nubilalis, in a novel range, North America, using MAXENT and CLIMEX modelling approaches. Results The models led to similar predictions about the expected distribution of the two species in Asia, and emphasized likely mismatches between host and natural enemy. Trichogramma ostriniae was predicted to occur in the summer corn region of China, with distribution limits linked to its sensitivity to cold, seasonality of radiation and precipitation. The modelled Ostrinia nubilalis distribution overlapped with the main corn production areas of the northeastern USA and Canada; temporary/seasonal suitable habitat was also predicted across the southeastern USA. Climate change scenarios are predicted to favour T. ostriniae over its hosts in northeastern China and North America. Conclusion The modelling approaches used here proved useful for assessing environmental factors linked to an egg parasitoid and its lepidopteran hosts and identifying areas potentially suitable for inundative releases. © 2017 Society of Chemical Industry.

Paper description: Internal transcribed spacer 2 (ITS2) sequences have proved to be useful in providing reliable identification of Trichogramma species. Identification of resident populations of Trichogramma wasps to species level has not been done in Serbia and has limited the potential for large scale use of these parasitoids. The current study used DNA sequences of the ITS2 gene to identify the most prevalent species in Vojvodina as Trischogramma brassicae . Accurate identification of native populations of Trichogramma wasps should help in the choice of the most suitable species for mass rearing and field release. Abstract: The European corn borer Ostrinia nubilalis (Hubner, 1796) (Lepidoptera: Pyralidae) is a major pest of corn ( Zea mays Linnaeus, 1753) in Serbia. Regular monitoring of O. nubilalis in the region of Kikinda (province of Vojvodina, Serbia) demonstrated a high percentage of parasitized eggs of this pest. Preliminary identification of the egg parasitoids based on morphological traits revealed that they belong to the genus Trichogramma Westwood, 1833 (Hymenoptera, Trichogrammatidae). The objective of this study was to identify the autochthonous Trichogramma species present in the region of Kikinda by using sequences of ribosomal DNA. Specimens of Trichogramma were reared from parasitized eggs of O. nubilalis collected at different localities in the region of Kikinda. The ITS2 gene was sequenced and subjected to BLAST searches of GenBank NCBI (National Center for Biotechnology Information). The majority of collected specimens were identified as Trichogramma brassicae Bezdenko, 1968, and a single specimen as T. evanescens Westwood, 1833. Intraspecific variation of T. brassicae was assessed by amplifying the mitochondrial cytochrome oxidase I (COI) gene . Wasps were also tested for the presence of the endosymbiotic bacteria Wolbachia Hertig, 1936 using Wolbachia -specific 16S and wsp primers as a preliminary test for evidence of thelytoky. Understanding of the diversity and biology of autochthonous Trichogramma species is a key step in the potential commercial implementation of these egg parasitoid wasps in the protection of corn fields in Serbia. https://doi.org/10.2298/ABS171103002I Received: November 13, 2017; Revised: January 12, 2018; Accepted: January 18, 2018; Published online: January 23, 2018 How to cite this article: Ivezic A, Rugman-Jones P, Stouthamer R, Ignjatovic-Cupina A. Molecular identification of Trichogramma egg parasitoids of Ostrinia nubilalis in northeastern Serbia. Arch Biol Sci. 2018;70(3):…

Background . Endosymbiotic bacteria of the genus Wolbachia are widespread in arthropods and often cause reproductive abnormalities in lepidopteran insects, including corn borers of the genus Ostrinia . Wolbachia - Ostrinia is a promising model for studies of parasite-host interactions yet parasite prevalence in natural insect host populations remains unknown. Materials and Methods . Molecular genetic screening and statistical analysis is applied to evaluate prevalence rates of Wolbachia in sympatric populations of two corn borer species. Individual genomic DNA samples were extracted from last instar larvae collected in nature from different forage plants. For each sample of DNA showing positive signal with insect-specific primers the detection is performed using three diagnostic loci of Wolbachia : 16SrRNA, gatB and fbpA. Results . Wolbachia -positive signal is obtained for 13.5% larvae of Ostrinia nubilalis (N = 141) and 31.9% larvae of Ostrinia scapulalis (N = 138). In different localities the Wolbachia prevalence ranged from 2.9% (N = 34) to 65.8% (N = 38). Significantly higher rates of Wolbachia prevalence in insects from mugwort and hemp ( O. scapulalis ) as compared to those from corn ( O. nubilalis ) are revealed in three out of four localities. Conclusions . Endosymbiotic bacteria of the genus Wolbachia are revealed in natural populations of corn borers for the first time for Eastern Europe. The prevalence rates can be high and this should be taken into consideration when reproductive isolation is examined in population of these hosts as well as establishment of laboratory cultures is performed.

Background. Microsporidia are ubiquitous parasites of animals, most abundant in arthropods and fishes. Many species of these parasites are important from standpoints of medicine, veterinary and agriculture. Microsporidium Nosema pyrausta is an important disease agent in corn borer populations causing adverse effect on host fitness. Genotyping of this parasite is necessary for proper species identification and intraspecific polymorphysm studies. Materials and Methods. Microsporidia-infected larvae of corn borers of the genus Ostrinia were recovered from Krasnodar Territory in Russia and Gomel Region in Belarus. Small subunit ribosomal RNA (SSU rRNA) was amplified and sequenced directly, while intergenic spacer (IGS) was amplified, cloned and sequenced (1-5 clones per sample) for four isolates of microsporidia. Sequences were aligned and compared using standard bioinformatics tools (Clustal W and BLAST). Results. SSU rRNA genotyping showed allocation of all four isolates to N. pyrausta with 100% identity to each other and 99.7% similarity to Nosema bombycis, the type species of the genus Nosema. High levels of IGS sequence variation (61-74%) is observed both between isolates of different species and populations of microsporidia as well as between molecular clones within parasite isolates from individual hosts. Conclusion. N. pyrausta is widespread in corn borer populations and its genetic structure is complicated, as in other species of these parasites. Further studies of molecular markers are needed for genetic differentiation of geographic isolates of N. pyrausta.

Prey have evolved a number of defenses against predation, and predators have developed means of countering these protective measures. Although caterpillars of the monarch butterfly,Danaus plexippus L., are defended by cardenolides sequestered from their host plants, the Chinese mantidTenodera sinensis Saussure guts the caterpillar before consuming the rest of the body. We hypothesized that this gutting behavior might be driven by the heterogeneous quality of prey tissue with respect to toxicity and/or nutrients. We conducted behavioral trials in which mantids were offered cardenolide-containing and cardenolide-freeD. plexippus caterpillars and butterflies. In addition, we fed mantids starved and unstarvedD. plexippus caterpillars from each cardenolide treatment and nontoxicOstrinia nubilalis Hübner caterpillars. These trials were coupled with elemental analysis of the gut and body tissues of bothD. plexippus caterpillars and corn borers. Cardenolides did not affect mantid behavior: mantids gutted both cardenolide-containing and cardenolide-free caterpillars. In contrast, mantids consumed bothO. nubilalis and starvedD. plexippus caterpillars entirely.Danaus plexippus body tissue has a lower C:N ratio than their gut contents, whileO. nubilalis have similar ratios; gutting may reflect the mantid’s ability to regulate nutrient uptake. Our results suggest that post-capture prey processing by mantids is likely driven by a sophisticated assessment of resource quality.

European corn borer Ostrinia nubilalis Hubner is one of the polyphagous insects which causes damage to more than 200 plant species. This study evaluated the applicability of using sterile insect technique (SIT) and F1 sterility (when male parents were irradiated by sub-sterilizing doses). Accordingly, the effect of gamma radiation at doses 100–350 Gy was determined on the biological and reproductive parameters.

Bacillus thuringiensis (Bt) Cry1Fa and Cry1Ab proteins are important Cry toxins due to their high, selective toxicity against a number of lepidopteran species, including important pests of corn and cotton. Competition binding assays are a classical tool for investigating Cry toxin interactions with target pest insects. We developed a fluorescence-based binding assay and assessed Cry1Fa and Cry1Ab toxin binding to brush border membrane preparations from lepidopteran corn pests including Ostrinia nubilalis (European corn borer, ECB), Diatraea grandiosella (south western corn borer, SWCB), and Helicoverpa zea (corn earworm, CEW). Homologous and heterologous competition binding assays with fluorophore-(Alexa488)-labeled Cry1Fa toxin showed that Cry1Fa shares binding site(s) with Cry1Ab toxin in ECB, and SWCB for which Cry1Ab has higher affinity than Cry1Fa. Apart from the shared binding sites, Cry1Ab and Cry1Fa bind an additional site(s) in ECB and SWCB. In CEW, Cry1Fa and Cry1Ab each, has a high affinity binding site(s), which binds the heterologous toxin with low affinity. The Cry1Ab-Cry1Fa toxin binding models for ECB, SWCB and CEW based on our results are considered in the context of what is known about acquired cross-resistance against Cry1Ab and Cry1Fa toxins.

Microsporidia are important natural regulators of many insect populations and as such are suitable microbial agents for the biological control of pest insects. Among the best-characterized microsporidia are those that infect the European corn borer, Ostrinia nubilalis. This chapter presents the analysis on Nosema pyrausta and Vairimorpha necatrix, as well as the recently described Nosema fur-nacalis. It examines the criteria in light of the need to recognize or develop more effective strains of microsporidia for corn borer control. The chapter reviews the advances in the identification of microsporidia that may facilitate the detection of introduced microsporidia and their delineation from indigenous species. It aims to evaluate what is known about microsporidian strains that offer different characteristics important to their use in biological control and the prospects for their manipulation via strain selection or genetic manipulation. Genetic studies and specific identification based on cytogenetic features are difficult because of the small size of microsporidia and their intracellular inhabitation.

Gatch, E. W., and Munkvold, G. P. 2002. Fungal species composition in maize stalks in relation to European corn borer injury and transgenic insect protection. Plant Dis. 86:1156-1162. The maize stalk rot complex is comprised of several fungal pathogens, including Gibberella zeae, Colletotrichum graminicola, Stenocarpella maydis, and several members of the genus Fusarium. The European corn borer (ECB) (Ostrinia nubilalis) can contribute to stalk rot development by creating entry wounds and by serving as a vector of some stalk rot pathogens, particularly Fusarium verticillioides. Transgenic insect protection of maize hybrids with insecticidal proteins derived from Bacillus thuringiensis greatly reduces ECB injury and may therefore alter the species composition and diversity of the stalk rot complex. Field experiments were conducted in 1998, 1999, and 2000 to compare the species composition and diversity of fungi infecting stalks of Bt and non-Bt maize hybrids. Hybrids representing five Bt types (or “events”) and their near-isogenic non-Bt counterparts were subjected to manual and natural infestations with ECB larvae. Stalk tissue samples were cultured to determine fungal species composition. At least one species was isolated from nearly every stalk and from both diseased and symptomless tissues. G. zeae was the most common species in 1998 and 1999, but C. graminicola was most common in 2000. The mean proportions of stalks infected with F. verticillioides, F. proliferatum, and F. subglutinans were significantly lower in Bt hybrids than in non-Bt hybrids in 2 of the 3 years. Conversely, the mean proportion of stalks infected with G. zeae was higher in some Bt hybrids than their non-Bt counterparts in two of the three years. F. verticillioides was more likely to be isolated from ECB-injured tissue, whereas G. zeae and C. graminicola were more likely to be isolated from tissue not associated with ECB injury. The overall species diversity of the stalk rot complex was lower in some Bt hybrids compared with their non-Bt counterparts in 1998 and 1999. ECB activity appeared to alter fungal species composition in stalks, reflecting the association between ECB injury and specific stalk rot pathogens, particularly F. verticillioides. The species composition of fungi infecting stalks of Bt hybrids differed from that of non-Bt hybrids, but the implications of this result are not yet clear.

The European corn borer (ECB), Ostrinia nubilalis Hubner (Lepidoptera: Crambidae), is a key pest of maize (Zea mays L.). In Northern Italy, larvae of the 2nd generation may damage cobs and kernels and the feeding activity can promote the proliferation of aflatoxigenic fungi, such as Aspergillus flavus, the major producer of aflatoxin B1 (AFB1). During 2017–2018, the efficacy of two strategies to control ECB in maize was assessed. Biological control strategy (Trichogramma brassicae Bezdenko and Bacillus thuringiensis Berliner), conventional chemical strategy (chlorantraniliprole) and an untreated control were compared to assess: i) the effect of the strategies on ECB infestation; ii) the association of ECB and AFB1 kernels contamination; iii) the impact of the strategies on beneficial arthropods. The conventional chemical strategy demonstrated the best control of ECB infestation, followed by biological control strategy and the untreated control. No significant differences in maize yield were found among strategies. The role of ECB on AFB1 concentration was demonstrated only in 2017, when higher level of infestations occurred simultaneously with an extended period of drought and high temperatures, sanctioning the important role of meteorological conditions on AFB1 contamination. The activity density of ground beetles, rove beetles and spiders and the mean number per leaf of the most abundant beneficial insects dwelling on plants (coccinellids, predatory thrips and lacewings) did not show significant changes between pre-and post-treatment with chlorantraniliprole, highlighting the selectivity of this pesticide in the short time. This study provides some contribution for the reduction of non-renewable input in Italian maize fields, demonstrating that biological control strategy, although less effective than conventional chemical control, can be a feasible approach to control ECB second larval generation, without any increment of AFB1 level in grains and yield loss.

To protect themselves from herbivory, plants have evolved an arsenal of physical and chemical defences and release a variety of volatile organic compounds (VOCs). By releasing these VOCs, a signalling plant can both reduce herbivory, sometimes by more than 90%, and also warn neighbouring plants about an attack. The aim of this study was to assess the influence of herbivory and insect extract application on VOC release by damaged/treated and nearby undamaged/untreated maize plants. We confirmed that European corn borer (Ostrinia nubilalis) larvae attack or larvae extract application induced maize VOC release. Greater amounts of (Z)-3-hexenal, (E)-2-hexenal, (Z)-3-hexen-1-ol, (E)-2-hexen-1-ol, β-myrcene, (Z)-3-hexen-1-yl acetate, 1-hexyl acetate, (Z)-ocimene, linalool, benzyl acetate, methyl salicylate, indole, methyl anthranilate, geranyl acetate, β-caryophyllene, (E)-β-farnesene and (Z)-3-hexenal, (Z)-3-hexen-1-ol, (Z)-3-hexen-1-yl acetate, (Z)-ocimene, linalool, indole, methyl anthranilate, geranyl acetate, β-caryophyllene and (E)-β-farnesene were released as a result of biotic stress after insect attack or insect extract application. The amounts of each VOC released were qualitatively and quantitatively distinct and dependent on time after biotic stress exposure. However, for all biotic stresses, significantly lower VOC induction was measured when leaves were damaged/treated for three days, as compared to seven days. Our work also demonstrated that undamaged/untreated neighbouring plants also release significant amounts of VOCs. This suggests that VOC emission by a damaged/treated plant stimulates VOC induction in nearby undamaged/untreated plants. However, the concentrations of all VOCs released by neighbouring undamaged/untreated maize plants were lower than those from damaged/treated plants and were negatively correlated with distance from a damaged/treated plant. Still, significant VOC induction occurred in undamaged/untreated plants even at 3 m distance from a damaged/infected plant. Our work suggests that maize plant protective defence responses (VOC emission) can be induced via application of European corn borer extracts.

Crops producing insecticidal toxins derived from the bacterium Bacillus thuringiensis (Bt) are widely planted to manage insect pests. Bt crops can provide an effective tool for pest management; however, the evolution of Bt resistance can diminish this benefit. The European corn borer, Ostrinia nubilalis Hubner, is a significant pest of maize and is widely managed with Bt maize in the Midwest of the United States. When Bt crops are grown in conjunction with non-Bt refuges, fitness costs of Bt resistance can delay the evolution of resistance. Importantly, fitness costs often vary with ecological factors, including host-plant genotype and diapause. In this study, we examined fitness costs associated with Cry1F resistance in O. nubilalis when insects were reared on three maize lines. Fitness costs were tested in two experiments. One experiment assessed the fitness costs when Cry1F-resistant and Cry1F-susceptible insects were reared on plants as larvae and experienced diapause. The second experiment tested resistant, susceptible and F1 heterozygotes that were reared on plants but did not experience diapause. Despite some evidence of greater adult longevity for Cry1F-resistant insects, these insects produced fewer fertile eggs than Cry1F-susceptible insects, and this occurred independent of diapause. Reduced fecundity was not detected among heterozygous individuals, which indicated that this fitness cost was recessive. Additionally, maize lines did not affect the magnitude of this fitness cost. The lower fitness of Cry1F-resistant O. nubilalis may contribute to the maintenance of Cry1F susceptibility in field populations more than a decade after Cry1F maize was commercialized.

The Cry1Fa protein from the bacterium Bacillus thuringiensis (Bt) is known for its potential to control lepidopteran pests, especially through transgenic expression in maize and cotton. The maize event TC1507 expressing the cry1Fa toxin gene became commercially available in the United States in 2003 for the management of key lepidopteran pests including the European corn borer, Ostrinia nubilalis, and the fall armyworm, Spodoptera frugiperda. A high-dose/refuge strategy has been widely adopted to delay evolution of resistance to event TC1507 and other transgenic Bt crops. Efficacy of this strategy depends on the crops expressing a high dose of the Bt toxin to targeted pests and adjacent refuges of non-Bt host plants serving as a source of abundant susceptible insects. While this strategy has proved effective in delaying O. nubilalis resistance, field-evolved resistance to event TC1507 has been reported in S. frugiperda populations in Puerto Rico, Brazil, and the southeastern United States. This paper examines available information on resistance to Cry1Fa in O. nubilalis and S. frugiperda and discusses how this information identifies opportunities to refine resistance management recommendations for Bt maize.

The European corn borer, Ostrinia nubilalis, is the most important pest insect of corn (Zea maize) in Germany, other European countries and historically in North America. However, in Europe only a few reports on naturally occurring biotic mortality factors are available. In the present review, an overview on pathogens including nematodes attacking O. nubilalis is compiled. The primary data were originally derived from long-term diagnostic studies on pathogens of arthropods carried out by the Institute for Biological Control, Darmstadt, Germany. These are updated with findings from the literature and supplemented with data from other pest insects of the same lepidopterous Crambidae family. The most important and most frequently found pathogens are the microsporidium Nosema pyrausta, the fungi Beauveria bassiana and to some extent also Fusarium spp., and the bacteria Bacillus thuringiensis and Serratia marcescens. Thus far, viruses have not been detected. Data on practical use of B. thuringiensis, B. bassiana and other fungi, N. pyrausta, and some nematodes for biological control of O. nubilalis are presented. Because of the importance of N. pyrausta as a population regulator of O. nubilalis, causing a chronic disease in larvae and adults, the current knowledge on the occurrence, biology, transmission, effects on the host insect, interactions with beneficial insects and field experiments is presented. These data are complemented with former, unpublished studies on the occurrence and incidence of N. pyrausta in some German and French O. nubilalis populations (Hessen, Baden-Wurttemberg, Alsace). The review clearly indicates that pathogens may play an important role as naturally occurring mortality factors of O. nubilalis which may also be used as biocontrol agents or within conservation biological control strategies.

Blending Bacillus thuringiensis (Bt) corn and non-Bt corn seeds in the same bag is a convenient and easier compliance solution to satisfy refuge regulations. However, there has been considerable debate and concern about larval movement of the target insects in a seed blend because of its potential effect on the development of resistance. Reported here are studies to determine the larval movement of European corn borers, Ostrinia nubilalis (Hubner), in a seed blend compared to a structured refuge and their effects on survival, feeding injury and larval fitness. Also evaluated is the relative performance of a purple-seeded corn hybrid as a surrogate host plant for tracking gene flow. Seed blend and structured refuge arrangements of Agrisure GT/CB/LL Bt corn expressing Cry1Ab and isoline plants were evaluated at two locations in 2013 and 2014 by manually infesting refuge plants with first and second generation corn borer larvae. The majority of leaf and stalk injury was recorded on the refuge plant, which amounted to 66–83% of the total tunnel length. In both refuge arrangements, plant injury and the number of larvae recovered significantly decreased on neighboring plants with increasing distance away from the refuge plant in the same row. The relative decline in injury was much more pronounced in the seed blend, with neighbor Bt plants experiencing very minor tunneling or no injury at all due to the high dose trait. Eighty-nine percent fewer live larvae were also found on neighbor Bt plants in the seed blend compared to those found in the structured refuge plots. Larvae that moved to Bt plants as well as older instars that were manually placed on Bt plants failed to complete development. Comparison of the number of tunnels and larvae recovered per refuge plant indicated that the seed blend may not produce as many susceptible individuals as those produced in a structured refuge arrangement. Although the purple-seeded hybrid showed lower levels of ear and stalk injury, it could serve as an appropriate surrogate refuge plant to track gene flow.

Understanding the behavior of pests targeted with Bacillus thuringiensis Berliner (Bt) crops is important to define resistance management strategies. Particularly the study of larval movement between plants is important to determine the feasibility of refuge configurations. Exposure to Bt maize, Zea mays L. (Poaceae), has been suggested to increase larval movement in lepidopteran species but few studies have examined the potential for resistance to interact with behavioral responses to Bt toxins. Choice and no-choice experiments were conducted with Spodoptera frugiperda (JE Smith) (Lepidoptera: Noctuidae) and Ostrinia nubilalis (Hubner) (Lepidoptera: Crambidae) to determine whether Cry1F resistance influences neonate movement. Leaf discs of Cry1F maize and the corresponding isoline were used to characterize behavioral responses. In both experiments, the location (on or off of plant tissues) and mortality of susceptible and Cry1F resistant neonates was recorded for 5 days, but the analysis of larvae location was performed until 7 h. Our results indicated no strong difference between resistant and susceptible phenotypes in S. frugiperda and O. nubilalis, although a small percentage of susceptible neonates in both species abandoned maize tissue expressing Cry1F. However, significant behavioral differences were observed between species. Ostrinia nubilalis exhibited increased movement between leaf discs, whereas S. frugiperda selected plant tissue within the first 30 min and remained on the chosen plant regardless of the presence of Cry1F. Spodoptera frugiperda reduced larval movement may have implications to refuge configuration. This study represents the first step toward understanding the effects of Cry1F resistance on Lepidoptera larval behavior. Information regarding behavioral differences between species could aid in developing better and more flexible resistance management strategies.

A series of new pymetrozine analogues containing both methyl on the imine carbon and phenoxy group at the pyridine ring were designed and synthesized. Their insecticidal activities against bean aphid (Aphis craccivora), mosquito larvae (Culex pipiens pallens), cotton bollworm (Helicoverpa armigera), corn borer (Ostrinia nubilalis) and oriental armyworm (Mythimna separata) were evaluated. The results of bioassays indicated that most of the target compounds showed good insecticidal activity against bean aphid; especially, IIIf (80%) and IIIl (80%) exhibited higher aphicidal activity than pymetrozine (30%) at 5mg/kg, and the two compounds still showed 20% and 30% mortality at 2.5mg/kg, respectively, whereas pymetrozine displayed no activity at the same concentration. These compounds exhibited a completely different structure-activity relationship to that of known pymetrozine derivatives, in which it is thought introducing alkyl group on the imine carbon could be detrimental to the activities. Our new result suggested that the methyl on the imine carbon and phenoxy group at the pyridine ring of phenoxy group may play additive effects on the improvement of aphicidal activity. Besides this, compound IIIs, containing an allyl at the para position of phenoxy group, exhibited excellent insecticidal activity against mosquito larvae, lepidoptera pests cotton bollworm, corn borer and oriental armyworm.

Sorghum (Sorghum bicolor (L.) Moench) brown midrib (bmr) mutant lines have reduced levels of lignin, which is a potentially useful trait for bioenergy production, but the effects of this trait on insect and plant pathogen interactions are unknown under field conditions. Field-grown bmr6, bmr12, and wild-type (WT) plants were examined for insect and disease damage. In most cases, observed frequency, population, or leaf area damage caused by insects or pathogens on bmr6 or bmr12 plants were not greater than those observed on WT plants in the field or laboratory assays. European corn borers [Ostrinia nubilalis (Hubner)(Lepidoptera: Pyralidae)] often caused lower amounts of leaf damage to bmr6 leaves compared to bmr12 and sometimes WT leaves in the field study. Leaf damage by corn earworms [Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae)] and fall armyworms [Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae)] in laboratory assays was often lower for bmr versus WT leaves. Incidence of disease lesions was significantly higher on bmr6 compared to WT plants for one of three samplings in 2011, but the opposite trend was observed overall in 2012 and no significant differences were noted in 2013. When corn earworms and fall armyworms were fed the excised pith, bmr6 and/or bmr12 pith caused significant morality to one or both insect species in all 3 years. Damage variability between the 3 years may have been due to hotter and drier than normal conditions in 2012. Thus, bmr lines of sorghum suitable for bioenergy production have potential for sustainable production in the field.

The wasp, Macrocentrus cingulum, is a polyembryonic endoparasitoid from the insect order Hymenoptera deployed successfully as biological control agent for the control of corn pest insect species from the Lepidopteran genus Ostrinia. As an insect, olfaction plays a crucial role for foraging, host-seeking, mating, ovipositing and avoiding toxic substances. Consequently, the molecular basis of olfaction in this wasp is of particular interest. In an effort to expand our understanding of chemosensory pathways, transcripts encoding the highly conserved insect odorant co-receptor (Orco) were isolated from this wasp. The McinOrco shares high degree of sequence homology with known Orco proteins in other insects. Real-time PCR experiments indicate that the McinOrco is expressed mainly in adult antennae, with expression levels differing by sex. The expressing cells were visualized by adapting a whole mount fluorescence in situ hybridization method. Orco-expressing sensory neurons were shown to be abundant in all 39 segments of male antenna. The highest densities of Orco-expressing cells were identified in segment 20, whereas segment 7 comprised in low level. In case of female antenna, levels of Orco-expressing cells were lower than male antenna. Almost expressed cells were distributed over 40 flagellomeres, with decreasing numbers towards the proximal end. These demonstrate the sexual dimorphism concerning the expression cells of Orco. We are the first to report, Orco in polyembryonic endoparasitoid M. cingulum, which play an important role in detection of olfactory cues in this wasp.

Hexokinase (HK) is a key enzyme in chitin biosynthesis in insects and plays an important role in development and energy regulation. It also performs a crucial role in the synthesis of Glucose-6-phosphate and its putative functions are studied via injection of dsRNA corresponding to the hexokinase gene from Cnaphalocrocis medinalis (CmHK). This study was designed to analyze the characteristics and expression patterns of HK-related genes in various tissues of C. medinalis at different developmental stages. The CmHK ORF is a 1359 bp in length, encoding a protein of 452 amino acids, with homology and cluster analysis showing that CmHK shares an 85.11% sequence similarity with hexokinase from Ostrinia furnacalis.CmHK was highly expressed in the ovary and in the fifth instar larvae. Injection of dsCmHK significantly suppressed mRNA expression (73.6%) 120 h post-dsRNA injection as compared to a control group. The results demonstrated an increased incidence of larval and pupal mortality of 80% and 78%, respectively, with significant variation in the sex ratio between males (68.33%) and females (35%), overt larval deformities, and a reduction in average weight gain observed 120 h post-dsRNA injection. In addition, dsCmHK-injected C. medinalis showed a significant reduction in ovulation per female and larval hatching rate, along with increased larval and pupal mortality and variation in male and female emergence over three generations (G1, G2, and G3). Taken together, the outcomes of the study provide a foundation to study gene function and a new dimension to control C. medinalis by transgenic RNAi technology.

Microsporidia are obligate intracellular parasites that affect the population density of many insect pests. In particular, infection with Nosema pyrausta is one of the major mortality factors for the European corn borer Ostrinia nubilalis, the Asian corn borer Ostrinia furnacalis and the adzuki bean borer Ostrinia scapulalis. The purpose of the work is to compare the susceptibility to N. pyrausta and pathogenesis of three species of moths of the genus Ostrinia. Studies conducted over 2 years have shown that in all three species of host insects under laboratory conditions, both during oral infection and transovarian transmission of infection (in the daughter generations of experimentally infected insects), only diplokaryotic spores formed corresponding to the main morphotype of the genus Nosema. Mean lethal time increased with instar of larvae used for infection but didn’t differ between the three species. The rates of transovarial transmission of N. pyrausta were also similar. Thus, all the insect species examined may equally participate in the parasite persistence in nature and serve as model laboratory hosts for parasitological research and mass propagation of the microsporidium.

Lepidopteran insects use sex pheromones for sexual communication. Pheromone receptors expressed on peripheral olfactory receptor neurons (ORNs) are critical part to detect the sex pheromones. In genus Ostrinia, several pheromone receptors were functional analyzed in O. nubilalis and O. scapulalis but the knowledge in O. furnacalis was rare. In this study, seven pheromone receptors were deorphanized by heterologous expression system of Xenopus oocytes. Functional types of sensilla trichoidea were classified by single sensillum recordings to interpret the response pattern of olfactory sensory neurons to Ostrinia pheromone components. OfurOR4 and OfurOR6 responded to the major sex pheromone Z/E12-14:OAc. OfurOR4 is the main receptor for both Z/E12-14:OAc and OfurOR6 mainly responded to E12-14:OAc. Functional differentiation of gene duplication were found between OfurOR5a and OfurOR5b. OfurOR5b showed a broad response to most of the pheromone components in O. furnacalis, whereas OfurOR5a was found without ligands. OfurOR7 showed a specific response to Z9-14:OAc and OfurOR8 mainly responded to Z11-14:OAc and E11-14:OAc. OfurOR3 did not respond to any pheromone components. Our results improved the current knowledge of pheromone reception in Ostrinia species which may contribute to speciation.

We present here a comprehensive analysis of the transcriptomic response to plant environments in ovipositing females of two sibling species of phytophagous moths affiliated to different host ranges: the European corn borer (ECB) and the adzuki bean borer (ABB). We first assembled and annotated a de novo reference transcriptome based on a high throughput RNA sequencing of females placed in different plant environments, then we measured differences in gene expression between ECB and ABB, and also within each moth species between environments. We further related the differentially expressed (DE) genes to the host preference in ECB and ABB and highlighted the functional categories involved. More specifically, we conducted an analysis on chemosensory genes previously characterized in ECB, ABB and other related Ostrinia species, as these genes are considered as good candidates for the host recognition before oviposition.Overall, we recorded more DE genes in ECB than in ABB samples, what could highlight the higher strength of the host specialization in ECB compared to ABB as observed at the behavioral level. We also noticed that the genes involved in the preference for their respective host were different between ECB and ABB. At the functional level, the response to plant environment in ECB and ABB during oviposition involved many processes, including the chemosensory repertoire as expected, but also metabolism of carbohydrates, lipids, proteins, and amino acids, detoxification mechanisms and immunity.All together, our results allowed identifying genes and functions candidates for specialization and also for the species divergence between ECB and ABB. By ad-hoc categorization, we discriminated some genes responding to the environment with similar or divergent pattern in ECB and ABB. Among them, we highlighted new lines of research like carbohydrates metabolism or virus and retrovirus dynamics.

We assessed the effectiveness of a biofortified maize line (4BtxHC) which accumulates high levels of antioxidant carotenoids that also expressed the insecticidal Cry1Ac Bacillus thuringiensis (Bt) gene against the European corn borer Ostrinia nubilalis. This line had been previously engineered to accumulate carotenoids specifically in the seed endosperm, whereas the Bt gene was expressed constitutively. The concentrations of Bt toxin (Cry 1Ac) in the leaves of the 4Bt and 4BtxHC lines were not significantly different at 47±6 μg/g of fresh weight (FW); neither were they in the kernels of both lines (35±3 μg/g FW). The kernels and leaves were toxic to the larvae of O. nubilalis. However, the insecticidal activity was substantially lower (ca. 20%) than that of lines that expressed only Bt in spite that the two lines showed a quantity of toxin not significantly different in kernels or in leaves. Although the reduced effectiveness of Cry1Ac in kernels may not be entirely surprising, the observation of the same phenomenon in vegetative tissues was unexpected. When semi-artificial diets containing kernels from 4Bt supplemented with different levels of β-carotene were used in insect bioassays, the β-carotene moderated the effectiveness of the Bt similarly to the plant material with carotenoid enrichment. To elucidate the biochemical basis of the reduced effectiveness of Bt toxin in the carotenoid-enriched plants, we measured the activity of three enzymes known to be implicated in the detoxification defence, namely, catalase, superoxide dismutase and glutathione S-transferase. Whereas Cry1Ac expression significantly increased SOD and CAT enzymatic activity in the absence of carotenoids, carotenoids, either in 4BtxHC or in artificial diets enriched with β-carotene, significantly lowered CAT activity. Carotenoids can therefore moderate the susceptibility of the maize borer O. nubilalis to Cry1Ac, and we hypothesize that their role as antioxidants could explain this phenomenon via their scavenging of reactive oxygen species produced during Cry1Ac detoxification in the larvae. The involvement of this mechanism in the decreased mortality caused by Cry1Ac when carotenoids are present in the diet is discussed. This work was supported by RecerCaixa PC084082 VitaMaize, funded by CaixaBank, to PC; Ministerio Economia y Competitividad AGL2011- 23966, AGL2014- 53970, to RA; and BIO2014- 54426-P, funded by the Spanish R&D Programme, to PC. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Paper description: Ostrinia nubilalis is one of the most damaging insect pests of maize with a significant impact on crop production. Measuring the differences in the effects of the insecticides on the antioxidative system helps in predicting the speed of insect adaptation to insecticide doses, which allows for timely substitution of the applied insecticides. This is the first examination of the effects of indoxacarb, chlorantraniliprole and the combination of chlorantranilprole and lambda cyhalothrin on the antioxidative system in a sample population of O. nubilalis from Serbia. Abstract: The European corn borer ( Ostrinia nubilalis ) is one of the most important insect pests of maize, and has a significant impact on the production of this crop. In this work, we examined the effects of different insecticides on the antioxidative defense system of O. nubilalis larvae. The experimental setup consisted of a completely randomized block design with 4 replicates. Four experimental groups were formed as follows: control (C), indoxacarb (250 mL ha -1 ), chlorantraniliprole (100 mL ha -1 ) and the chlorantraniliprole+lambda cyhalothrin (200 mL ha -1 ) group. Larvae from maize stems were collected 20 days after insecticide application and the whole larvae were homogenized. The activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR) and glutathione S-transferase (GST), and the total amount of free SH groups were assessed. Comparison of the experimental groups showed that indoxacarb significantly affected the activities of GST, GPx and the total amount of free SH groups, while chlorantraniliprole significantly affected the activities of SOD, CAT, GST and the total amount of free SH groups, while chlorantraniliprole+lambda cyhalothrin significantly affected the activities of CAT, GST and the total amount of free SH groups. The results show that exposure to insecticides considerably affects the antioxidative defense components of the European corn borer larvae, especially chlorantraniliprole (T2). https://doi.org/10.2298/ABS180701042F Received: July 1, 2018; Revised: September 1, 2018; Accepted: September 10, 2018; Published online: October 2, 2018 How to cite this article: Franeta F, Mircic D, Todorovic D, Milovac Ž, Granica N, Obradovic S, Peric-Mataruga V. Effects of different insecticides on the antioxidative defense system of the European corn borer ( Ostrinia nubilalis Hubner) (Lepidoptera: Crambidae) larvae. Arch Biol Sci. 2018;70(4):765-73.

Throughout an organism's lifetime, resources are strategically allocated to many different functions, including reproduction. Reproduction can be costly for both sexes; females produce nutrient-rich eggs, whereas males of many species produce large and complex ejaculates. In capital breeding insects, nutrients are mainly acquired during the larval period, yet allocation decisions impact the reproductive fitness of adults. The present study examines the effect of larval dietary nitrogen on both male and female reproductive traits in the European corn borer moth Ostrinia nubilalis Hubner, whose adults do not feed and whose males transfer a large, nitrogen-rich spermatophore. One day post-eclosion, O. nubilalis adults reared on one of three different diets (3.0%, 1.6%, or 1.1% nitrogen) are mated and two experiments are undertaken: one to measure nitrogen and carbon content of male ejaculates, and the other to determine female fecundity and fertility. Although male larval diet does not alter the percentage nitrogen content of adult somatic tissue, males reared on the higher nitrogen diet (3.0%) produce spermatophores with increased nitrogen relative to somatic nitrogen. Furthermore, females raised on the 3.0% nitrogen diet receive spermatophores with lower carbon : nitrogen ratios and thus more nitrogen. Overall, females lay more eggs as their larval dietary nitrogen increases, although they lay fewer eggs when their mates are raised on the higher (3.0%) nitrogen diet. This suggests that O. nubilalis females may use male-derived nitrogen not to supplement egg production, but rather for somatic maintenance. Overall, the present study furthers our understanding of how larval diet can affect adult fitness in Lepidoptera.

A significant feature of the genomes of Lepidoptera, butterflies and moths, is the high conservation of chromosome organization. Recent remarkable progress in genome sequencing of Lepidoptera has revealed that syntenic gene order is extensively conserved across phylogenetically distant species. The ancestral karyotype of Lepidoptera is thought to be n=31; however, that of the most well-studied moth, Bombyx mori, is n=28, and diverse studies suggest that three chromosomal fusion events occurred in this lineage. To identify the boundaries between predicted ancient fusions involving B. mori chromosomes 11, 23 and 24, we constructed fluorescence in situ hybridization (FISH)-based chromosome maps of the European corn borer, Ostrinia nubilalis (n=31). We first determined a 511 Mb genomic sequence of the Asian corn borer, O. furnacalis, a congener of O. nubilalis, and isolated bacterial artificial chromosomes and fosmid clones that were expected to localize in candidate regions for the boundaries using these sequences. Combined with FISH and genetic analysis, we narrowed down the candidate regions to 40 kb–1.5 Mb, in strong agreement with a previous estimate based on the genome of a butterfly, Melitaea cinxia. The significant difference in the lengths of the candidate regions where no functional genes were observed may reflect the evolutionary time after fusion events.

Field evolved resistance of insect populations to Bacillus thuringiensis (Bt) crystalline (Cry) toxins expressed by crop plants has resulted in reduced control of insect feeding damage to field crops, and threatens the sustainability of Bt transgenic technologies. A single quantitative trait locus (QTL) that determines resistance in Ostrinia nubilalis larvae capable of surviving on reproductive stage transgenic corn that express the Bt Cry1Fa toxin was previously mapped to linkage group 12 (LG12) in a backcross pedigree. Fine mapping with high-throughput single nucleotide polymorphism (SNP) anchor markers, a candidate ABC transporter (abcc2) marker, and de novo mutations predicted from a genotyping-by-sequencing (GBS) data redefined a 268.8 cM LG12. The single QTL on LG12 spanned an approximate 46.1 cM region, in which marker 02302.286 and abcc2 were ≤2.81 cM, and the GBS marker 697 was an estimated 1.89 cM distant from the causal genetic factor. This positional mapping data showed that an O. nubilalis genome region encoding an abcc2 transporter is in proximity to a single QTL involved in the inheritance of Cry1F resistance, and will assist in the future identification the mutation(s) involved with this phenotype.