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The movement of miniature inverted repeat transposable elements (MITEs) modifies genome structure and function. We describe the microsatellite-associated interspersed nuclear element 2 (MINE-2), that integrates at consensus WTTTT target sites, creates dinucleotide TT target site duplications (TSDs), and forms predicted MITE-like secondary structures; a 5' subterminal inverted repeat (SIR; AGGGTTCCGTAG) that is partially complementary to a 5' inverted repeat (IR; ACGAAGCCCT) and 3'-SIRs (TTACGGAACCCT). A (GTCY)(n) microsatellite is hitchhiking downstream of conserved 5'MINE-2 secondary structures, causing flanking sequence similarity amongst mobile microsatellite loci. Transfection of insect cell lines indicates that MITE-like secondary structures are sufficient to mediate genome integration, and provides insight into the transposition mechanism used by MINE-2s., (© 2010 The Authors. Insect Molecular Biology © 2010 The Royal Entomological Society.)

The behavior of pests targeted by Bacillus thuringiensis (Bt) crops has been recognized as an important factor to define resistance management plans. However, most data do not include the possible impact resistance may have on the behavior of pests. To examine whether resistance influences behavior of European corn borer, Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae), neonates after exposure to dietary Bt, the responses of Cry1Ab-resistant, -susceptible, and hybrid (F1) lines from two populations were compared in laboratory tests by using artificial diet mixed with 10-50% Cry1Ab or non-Bt isoline corn, Zea mays L., tissue. In no-choice tests, resistant (and usually hybrid) lines were less likely to be irritated (i.e., to move away after physical contact with diet containing Cry1Ab) than susceptible larvae after exposure to diets containing 10-50% Cry1Ab leaf tissue. Early in the no-choice tests (8 h), neonate O. nubilalis also were more likely to move off of diets that contained 10% non-Bt tissue compared with diets with 25 or 50% non-Bt tissue. In agreement with results from no-choice tests, choice tests with 10 or 25% tissue indicated that resistant (and sometimes hybrid) larvae were more likely than susceptible neonates to be found on diet with Cry1Ab. For choice tests, differences among lines seemed dependent on the amount of Cry1Ab tissue incorporated into diets. Results suggest differences in behavior are a result of reduced physiological susceptibility to Cry1Ab and are not an independent behavioral component to resistance.

Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae) is one of the most damaging insect pests of corn. Studies were conducted to determine whether live larval counts obtained from corn whorls were predictive of the amount of larval tunneling that would result in the stalk of the plant 40 d later at the end of larval development. Whorls from plants treated with Dipel 10G (6,400 IU per whorl) and untreated controls, both infested with O. nubilalis neonates, were evaluated for the number of live larvae in 50 whorls 5, 7, 9, and 12 d after Bacillus thuringiensis (Berliner) (Bacillales: Bacillaceae) application. Forty days after larval infestation, 25 plants from each plot were split from tassel to base, and the length of larval tunneling was recorded. There was a strong relationship between numbers of live larvae in the plant whorl and the length of larval tunneling that resulted. While linear at each location, there was significant variation in the relationship among locations, indicating that comparisons could not be made between years or locations within a year. Blocks within a location on a given year did not vary significantly and reproducible results were obtained each year within a given location as well as on any of the whorl pulling dates evaluated. Because of its ease of use, predictability, and rapid return of results, we propose this technique as an additional method to shot-hole feeding and stalk splitting to evaluate the effectiveness of O. nubilalis management strategies.

Genes expressed in lepidopteran midgut tissues are involved in digestion and Bacillus thuringiensis (Bt) toxin resistance traits. Five hundred and thirty five unique transcripts were annotated from 1745 high quality O. nubilalis larval midgut expressed sequence tags (ESTs). Full-length cDNA sequence of 12 putative serine proteinase genes and 3 partial O. nubilalis aminopeptidase N protein genes, apn1, apn3, and apn4, were obtained, and genes may have roles in plant feeding and Bt toxin resistance traits of Ostrinia larvae. The EST library was not normalized and insert frequencies reflect transcript levels under the initial treatment conditions and redundancy of inserts from highly expressed transcripts allowed prediction of putative single nucleotide polymorphisms (SNPs). Ten di-, tri- or tetranucleotide repeat unit microsatellite loci were identified, and minisatellite repeats were observed within the C-termini of two encoded serine proteinases. Molecular markers showed polymorphism at 28 SNP loci and one microsatellite locus, and Mendelian inheritance indicated that markers were applicable to genome mapping applications. This O. nubilalis larval midgut EST collection is a resource for gene discovery, expression information, and allelic variation for use in genetic marker development.

Genetically engineered corn hybrids that contain a cry gene from the bacterium Bacillus thuringiensis Berliner (Bt) are gaining popularity for controlling the corn pest Ostrinia nubilalis (Hübner). Continuous use of Bt corn, however, could select for O. nubilalis that are resistant to this corn. Monitoring for insect resistance is important, because it could help maintain the Bt technology. A possible monitoring method is to collect larval insects in commercial drying bins after harvest from Bt seed production fields. A drawback to this method is that these collections may be contaminated by insects that moved as later instars from severed non-Bt male rows into the adjacent Bt female rows. These larvae have little to no exposure to Bt toxin, resulting in possible "false positives." The objectives of this study were to first find which combination of planting and severing dates produces the least number of larvae that move from non-Bt male plants to Bt female plants and to assess O. nubilalis larval movement from severed non-Bt male rows to Bt female rows. Field studies in 2002 and 2003 were designed to simulate a hybrid seed production field. Results suggest that movement of O. nubilalis larvae from male corn is minimized when corn is planted early and male plants are severed by 2 wk post-anthesis. This reduces the likelihood of false positives by reducing the number of susceptible larvae moving between Bt and non-Bt plants. Also, larvae moved to all four female rows that were adjacent to the severed rows, but there were significantly more larvae found in the closest row compared with the other three. These results could be used to develop a monitoring program to find O. nubilalis larvae with resistance to Bt corn in field populations of O. nubilalis.

Transgenic corn, Zea mays L., hybrids expressing crystal protein endotoxin genes from Bacillus thuringiensis Berliner are an increasingly popular tactic for managing the European corn borer, Ostrinia nubilalis (Hübner), in North America. O. nubilalis populations also are often vulnerable to the ubiquitous entomopathogenic microsporidium Nosema pyrausta (Paillot). We examined the effect of feeding meridic diet incorporated with purified Cry1Ab on growth, development, and survival of Nosema-infected and uninfected neonate O. nubilalis. Infected larvae developed more slowly than uninfected larvae. Increasing the concentration of Cry1Ab in diet reduced larval development, and this effect was amplified by microsporidiosis. Infected larvae weighed significantly less than uninfected larvae. The relationship among Nosema infection, Cry1Ab concentration, and larval weight was fitted to an exponential function. The LC50 of infected larvae was one-third that of uninfected larvae, indicating that infected larvae are more vulnerable to toxin. This work has implications for resistance management of O. nubilalis and demonstrates that it is important to determine whether N. pyrausta is present when testing susceptibility of larvae to transgenic corn hybrids.

Differences in larval survival and development of bollworm, Helicoverpa zea (Boddie), and fall armyworm, Spodoptera frugiperda (J. E. Smith), respectively, were found to exist among commercially available Cry1A(c) transgenic Bacillus thuringiensis Berliner (Bt) varieties. Using a quantification assay (ELISA) to measure the levels of delta-endotoxin in two of these varieties ('DP 451B/RR' and 'NuCOTN 33B'), differences in the amount of delta-endotoxin present in various plant parts was correlated with larval survival of bollworms and larval development of fall armyworms throughout the growing season. Larvae that were fed on DP 451B/RR completed development faster and exhibited better survivorship than those larvae fed NuCOTN 33B, whereas lower levels of delta-endotoxin were generally detected in plant parts from DP 451B/RR compared with NuCOTN 33B. These differences may impact population dynamics of these pests which may be a critical factor in managing resistance to Bt. Furthermore, the utility of using this system for providing information to the grower concerning varietal choices may be more common in the future.