Development of diagnostic concentrations for monitoring Bacillus thuringiensis resistance in European corn borer (Lepidoptera: Crambidae)

Two candidate diagnostic concentrations of the Cry1Ab and Cry1Ac toxins from Bacillus thuringiensis corresponding to the LC99 and EC99 (effective concentration that causes 99% growth inhibition) for European corn borer, Ostrinia nubilalis (Hübner), were determined based on previously obtained baseline data. Validation experiments using field-collected European corn borer populations from across North America showed that for Cry1Ab, a concentration corresponding to the upper limit of the 95% confidence interval of the LC99, produced mortality > 99% for all populations tested. However, for Cry1Ac, adjustments and further validation are probably necessary. Development of B. thuringiensis resistance monitoring programs that rely on diagnostic techniques are discussed.     

Extended monitoring of resistance to Bacillus thuringiensis Cry1Ab maize in Diatraea saccharalis (Lepidoptera: Crambidae)

The sugarcane borer, Diatraea saccharalis (F.), is a major target of transgenic maize expressing Bacillus thuringiensis (Bt) proteins in South America and the mid-Southern region of the United States. During 2007-2009, a total of 986 feral individuals of D. saccharalis were collected from maize fields in six locations of Louisiana and Mississippi and examined for resistance to Cry1Ab maize using F 1/F 2 screens. Major resistance alleles to Cry1Ab maize in the populations sampled from non-Bt maize plants during 2007 and 2008 in Louisiana and 2009 in Mississippi were rare. From a total of 487 individuals collected from three locations in Louisiana in 2007 and 2008, only one individual was identified with major resistance alleles. In addition, no major resistance alleles were detected in 242 individuals collected from three locations in Mississippi in 2009. The frequency of major resistance alleles was estimated to be 0.002 with a 95% CI of 0.00025-0.0057 for the Louisiana populations and < 0.0061, with 95% probability, for the Mississippi populations. The resistance frequency estimated for the Louisiana populations in 2007 and 2008 was not significantly different from those reported previously for populations sampled in 2004-2006. However, among 200 individuals sampled from non-Bt maize plants in 2009 in Louisiana, six individuals were identified to possess major resistance alleles. The estimated major resistance allele frequency for the populations sampled from non-Bt maize plants in 2009 in Louisiana was 0.0176 with a 95% CI of 0.0072 to 0.0328, which was significantly greater than those estimated for the populations collected in 2004-2008. Similarly, the frequency of minor resistance alleles to Cry1Ab maize for the Louisiana populations collected in 2009 was also significantly greater than those estimated for the populations sampled before. In addition, two out of 57 feral individuals collected from Bt maize plants in Louisiana in 2009 were identified to carry major resistance alleles to Cry1Ab maize. Since 2010, transgenic maize expressing pyramided Bt genes has been planted in the US mid-Southern region and by 2011, pyramided Bt maize has replaced Cry1Ab maize as the dominant Bt maize for managing lepidopteran pests including D. saccharalis. The timely switching from single-gene Cry1Ab maize to the pyramided Bt maize should prevent further increases in Cry1Ab resistance allele frequency and thus ensure the continued success of Bt maize for managing D. saccharalis in the region.     

Evaluation of transgenic Bacillus thuringiensis corn hybrids against Cry1Ab-susceptible and -resistant sugarcane borer (Lepidoptera: Crambidae)

A Louisiana strain of the sugarcane borer, Diatraea saccharalis (F.) (Lepidoptera: Crambidae), was selected for resistance to the CrylAb protein of Bacillus thuringiensis (Bt) by using an F2 screening procedure. Survival of Bt-resistant, -susceptible, and -heterozygous genotypes of sugarcane borer was evaluated on vegetative and reproductive stages of five non-Bt and seven Bt field corn, Zea mays L., hybrids in a greenhouse study. Larval survival was recorded 21 d after infestation of neonates on potted plants. Larval survival across the three sugarcane borer genotypes and five non-Bt corn hybrids after 21 d ranged from 23.6 +/- 5.2% (mean +/- SEM) to 57.5 +/- 5.2%. Mean survival of Cry1Ab-resistant larvae on vegetative and reproductive plant stages was 12 and 21%, respectively. During the vegetative stages, all seven Bt corn hybrids were highly efficacious against Cry1Ab-susceptible and -heterozygous genotypes of sugarcane borer, with a larval survival rate of <2% for the Bt-susceptible genotype and < or =5% for the heterozygotes. However, 8-18% of the heterozygous genotype survived on reproductive stage plants for four of the seven Bt corn hybrids tested. The variation in performance of Bt corn cultivars at vegetative and reproductive growth stages against Cry1Ab resistant sugarcane borer suggests differential seasonal expression that may hasten resistance in the field. Bt corn hybrids expressing a "high dose" for European corn borer, Ostrinia nubilalis (Hübner), may not produce a sufficient high dose for the sugarcane borer.     

Management of feeding damage and survival of southwestern corn borer and sugarcane borer (Lepidoptera: Crambidae) with Bacillus thuringiensis transgenic field corn

The efficacy of transgenic corn hybrids expressing an insecticidal Bacillus thuringiensis (Bt) delta-endotoxin from different transformation events was evaluated in field corn, Zea mays L., against the southwestern corn borer, Diatraea grandiosella Dyar, and sugarcane borer, Diatraea saccharalis (F.). Susceptibilities of neonates and third instars were determined on Bt and non-Bt corn plants (V6 and R1 stages) in field plots and corn leaf tissue feeding exposure in laboratory bioassays. Bt corn hybrids associated with MON810 and CBH351 transformation events sustained significantly less injury by southwestern corn borer and sugarcane borer during mid-whorl stage infestations compared with their respective non-Bt hybrid equivalents. Southwestern corn borer and sugarcane borer feeding injury to ear leaf-sheath and husk tissues during the silking stage of corn was significantly reduced in MON810 and CBH351 Bt corn compared with their respective non-Bt hybrids. However, resistance levels to feeding injury in Bt hybrids associated with the MON810 event were significantly higher than that in the hybrid associated with the CBH351 event. Southwestern corn borer and sugarcane borer caused more feeding injury to husk tissue than to ear leaf-sheath tissue in both Bt and non-Bt hybrids infested during the silking stage. Laboratory performance of the MON810 event against southwestern corn borer and sugarcane borer varied among hybrids associated with the same event. Third instars of southwestern corn borer were highly susceptible to MON810 Bt corn hybrids in leaf tissue experiments. However, sugarcane borer larvae were susceptible to the MON810 event only in one of the Bt hybrids evaluated. Sugarcane borer mortality was significantly lower after 96 h of feeding exposure on CBH351 Bt corn leaf tissue than on MON810 Bt corn leaf tissue. Plant resistance to southwestern corn borer and sugarcane borer increased as plants matured, independent of the presence of a Bt construct. These results are essential to estimate the importance of Bt transgenic corn in areas of southern United States and other areas where mixed populations of southwestern corn borer and sugarcane borer are predominant and cause severe damage to corn production.     

Allele frequency of resistance to Bacillus thuringiensis Cry1ab corn in Louisiana populations of sugarcane borer (Lepidoptera: Crambidae)

Transgenic Bacillus thuringiensis (Bt) corn, Zea mays L., has been widely used to manage a corn borer complex in the mid-southern region of the United States. The sugarcane borer, Diatraea saccharalis (F.) (Lepidoptera: Crambidae), has become a dominant cornstalk boring species in some areas of this region, especially in Louisiana. Therefore, management of sugarcane borer resistance to Bt corn is critical to ensure the long-term sustainability of Bt corn for the region. This study screened 280 two-parent family-lines of sugarcane borer from four geographical populations in Louisiana during 2005 to determine whether Bt resistance allele frequency in sugarcane borer is sufficiently low to meet the rare resistance assumption of the current "high dose/refuge" resistance management strategy for Bt corn. These sugarcane borer family-lines were examined for Bt resistance by using novel F2 screening procedures. No major Bt resistance alleles were detected in these four populations. The estimated frequency of major Bt resistance alleles was < 0.0027, with a 95% probability and a detection power of 94%. The estimated minor resistance allele frequency was 0.0063, with a 95% CI of 0.0025-0.0117. During a previous study, a major Bt resistance allele was detected in one individual from 213 family-lines of another Louisiana population of sugarcane borer. Combining these data with the current screen, the frequency of major Bt resistance alleles across the five populations was 0.001, with a 95% credibility interval of 0.0001-0.0028 and a detection power of 95%. Major Bt resistance allele frequencies in Louisiana sugarcane borer populations seem to be low, and they should support the rare resistance allele requirement of the high dose/refuge strategy.     

Effect of environment on resistance to the European corn borer (Lepidoptera: Crambidae) in maize

The European corn borer, Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae), is a major pest of maize, Zea mays L., in many temperate parts of the world. Genotype-by-environment interaction effects can make relative performance unpredictable and may hamper selection for resistance to European corn borer. The objective of this study was to determine the effect of environment on genotypic reaction to European corn borer resistance in maize. A set of 12 maize inbred lines was chosen to represent a range of European corn borer responses. Eleven testing environments ranged from Delaware, Ohio, Illinois, Iowa, Nebraska, Missouri, to Mississippi. For length of stalk tunneling, environmental and genotypic main effects (estimated by restricted maximum likelihood) were >20- and 10-fold larger than their interaction effect, respectively. Length of tunneling means for genotypes (across environments) ranged from 10.1 to 35.4 cm. Several putatively resistant genotypes grouped with the susceptible checks, B73 and Mol7. By breaking factors and the interaction into single degree of freedom components, we observed that GEMS-0001 had significant crossover interactions toward less susceptibility in both Mississippi and the Nebraska environments. Environments displaying several crossover interactions indicated that European corn borer screening at these sites would not necessarily apply to other locations, whether due to small differences in experimental conduct and/or environmental effects. The five most resistant genotypes were fairly consistent across environments. Because all environments except Illinois used larvae from the same insectary, and these environments differed in damage intensity and rankings, it is unlikely that insect biotype was a factor contributing to genotype-by-environment effects.     

Susceptibility of Cry1Ab-resistant and -susceptible sugarcane borer (Lepidoptera: Crambidae) to four Bacillus thuringiensis toxins

Sugarcane borer, Diatraea saccharalis (F.), is a primary corn stalk borer pest targeted by transgenic corn expressing Bacillus thuringiensis (Bt) proteins in many areas of the mid-southern region of the United States. Recently, genes encoding for Cry1A.105 and Cry2Ab2 Bt proteins were transferred into corn plants (event MON 89034) for controlling lepidopteran pests. This new generation of Bt corn with stacked-genes of Cry1A.105 and Cry2Ab2 will become commercially available in 2009. Susceptibility of Cry1Ab-susceptible and -resistant strains of D. saccharalis were evaluated on four selected Bt proteins including Cry1Aa, Cry1Ac, Cry1A.105, and Cry2Ab2. The Cry1Ab-resistant strain is capable of completing its larval development on commercial Cry1Ab-expressing corn plants. Neonates of D. saccharalis were assayed on a meridic diet containing one of the four Cry proteins. Larval mortality, body weight, and number of surviving larvae that did not gain significant weight (<0.1 mg per larva) were recorded after 7 days. Cry1Aa was the most toxic protein against both insect strains, followed in decreasing potency by Cry1A.105, Cry1Ac, and Cry2Ab2. Using practical mortality (larvae either died or no significant weight gain after 7 days), the median lethal concentration (LC₅₀) of the Cry1Ab-resistant strain was estimated to be >80-, 45-, 4.1-, and −0.5-fold greater than that of the susceptible strain to Cry1Aa, Cry1Ac, Cry1A.105 and Cry2Ab2 proteins, respectively. This information should be useful to support the commercialization of the new Bt corn event MON 89034 for managing D. saccharalis in the mid-southern region of the United States.     

Conventional resistance of experimental maize lines to corn earworm (Lepidoptera: Noctuidae), fall armyworm (Lepidoptera: Noctuidae), southwestern corn borer (Lepidoptera: Crambidae), and sugarcane borer (Lepidoptera: Crambidae)

Plant resistance is a useful component of integrated pest management for several insects that are economically damaging to maize, Zea mays L. In this study, 15 experimental lines of maize derived from a backcross breeding program were evaluated for resistance to corn earworm, Helicoverpa zea (Boddie); fall armyworm, Spodoptera frugiperda (J. E. Smith); southwestern corn borer, Diatraea grandiosella Dyar; and sugarcane borer, Diatraea saccharalis (F.). Experimental line 100-R-3 was resistant in the field to leaf feeding by fall armyworm and line 116-B-10 was resistant in the field to leaf feeding by fall armyworm and leaf and stalk feeding by southwestern corn borer. When corn earworm larvae were fed field harvested silks from experimental line 81-9-B in the laboratory, their pupal weights were significantly lower than the pupal weights of larvae that were fed silks from the resistant control, Zapalote Chico. Maysin levels lower than those commonly associated with corn earworm resistance were present in the resistant experimental line, 107-8-7, indicating a new basis confers resistance to corn earworm in this line. These resistant experimental lines will provide plant breeders with new sources of resistance to lepidopterous insects for the development of improved maize breeding populations.     

Frequency of resistance to Bacillus thuringiensis toxin Cry1Ab in an Iowa population of European corn borer (Lepidoptera: Crambidae)

The refuge plus high-dose strategy for resistance management assumes that the frequency of resistance alleles is low. We used an F2 screen to estimate the frequency of resistance to transgenic corn that produces Bacillus thuringiensis Berliner Cry1Ab toxin (Bt corn) in an Iowa population of European corn borer, Ostrinia nubilalis (Hübner). We also proposed a modification to the statistical analysis of the F2 screen that extends its application for nonuniform prior distributions and for repeated sampling of a single population. Based on a sample of 188 isofemale lines derived from females caught at light traps during the 2nd flight of 1997, we show with 95% confidence that the frequency of resistance to Bt corn was <3.9 x 10(-3) in this Iowa population. These results provide weak evidence that the refuge plus high-dose strategy may be effective for managing resistance in O. nubilalis to Bt corn. Partial resistance to Cry1Ab toxin was found commonly. The 95% CI for the frequency of partial resistance were [8.2 x 10(-4), 9.4 x 10(-3)] for the Iowa population. Variable costs of the method were 14.90 dollars per isofemale line, which was a reduction of 25% compared with our initial estimate.     

Chronic exposure of the European corn borer (Lepidoptera: Crambidae) to Cry1Ab Bacillus thuringiensis toxin.

Transgenic corn expressing the insecticidal toxin from Bacillus thuringiensis Berliner is gaining support as an effective control technology for use against lepidopteran pests, particularly European corn borer, Ostrinia nubilalis Hübner (Lepidoptera: Crambidae). However, there is concern that widespread adoption of transgenic plants will rapidly lead to B. thuringiensis toxin resistance. Thus, long-term selection of O. nubilalis populations with the Cry1Ab B. thuringiensis toxin has been undertaken in several laboratories in the United States and in Europe. We present results from two independent selection experiments performed in laboratories at the University of Nebraska and at the Institut National de la Recherche Agronomique in France. Although the protocols and methods used by the two laboratories were different, the results were comparable. The highest level of resistance occurred at generation 7 (14-fold), generation 9 (13-fold), and generation 9 (32-fold) for three different strains. For each strain, the level of resistance fluctuated from generation to generation, although there were consistently significant decreases in toxin susceptibility across generations for all selected strains. These results suggest that low levels of resistance are common among widely distributed O. nubilalis populations.     

Sugarcane borer (Lepidoptera: Crambidae) management threshold assessment on four sugarcane cultivars

This research assesses the potential for using different economic injury thresholds in management of a key insect pest on susceptible and resistant commercially produced cultivars of sugarcane (Saccharum spp. hybrids). In a 2-yr sugarcane borer, Diatraea saccharalis (F.) (Lepidoptera: Crambidae), study involving four commercially produced sugarcane cultivars and four insecticide treatment thresholds, 'LCP 85-384' and 'HoCP 91-555' were the most susceptible based on percentage of bored internodes compared with the more resistant 'HoCP 85-845' and 'CP 70-321'. In 2001, the 10% infested stalks threshold was not as effective as the 5% early season-10% late season and 5% full season for HoCP 91-555. Based on D. saccharalis injury under natural infestation conditions, susceptible cultivars seem to require a lower infestation threshold than the more resistant cultivars to achieve adequate injury reduction. Among yield components, only the theoretical recoverable sugar per stalk was significantly increased by applying insecticides. With the resistant HoCP 85-845, differences were not detected for percentage of bored internodes among treated versus untreated management regimes. The resistant HoCP 85-845 had higher levels of fiber in our study; however, no clear pattern on resistance mechanisms was established, because the resistant cultivar CP 70-321 had comparatively low levels of fiber. The development of cultivar-specific thresholds is expected to lower the amount of insecticide used for D. saccharalis management in the sugarcane industry, reduce selection pressure, and delay the development of insecticide resistance.     

Genetic basis of resistance to fall armyworm (Lepidoptera: Noctuidae) and southwestern corn borer (Lepidoptera: Crambidae) leaf-feeding damage in maize

Leaf-feeding damage by first generation larvae of fall armyworm, Spodopter frugiperda (J. E. Smith) (Lepidoptera: Noctuidae), and southwestern corn borer, Diatraea grandiosella Dyar (Lepidoptera: Crambidae), cause major economic losses each year in maize, Zea mays L. A previous study identified quantitative trait loci (QTL) contributing to reduced leaf-feeding damage by these insects in the maize line Mp704. This study was initiated to identify QTL and their interactions associated with first generation leaf-feeding damage by fall armyworm and southwestern corn borer. QTL associated with fall armyworm and southwestern corn borer resistance in resistant line Mp708 were identified and compared with Mp704. Multiple trait analysis (MTA) of both data sets was then used to identify the most important genetic regions affecting resistance to fall armyworm and southwestern corn borer leaf-feeding damage. Genetic models containing four and seven QTL explained southwestern corn borer and fall armyworm resistance, respectively, in Mp708. Key genomic regions on chromosomes 1, 5, 7, and 9 were identified by MTA in Mp704 and Mp708 that confer resistance to both fall armyworm and southwestern corn borer. QTL regions on chromosomes 1, 5, 7, and 9 contained resistance to both insects and were present in both resistant lines. These regions correspond with previously identified QTL related to resistance to other lepidopteran insects, suggesting that broad-spectrum resistance to leaf feeding is primarily controlled by only a few genetic regions in this germplasm.     

F2 screen for resistance to a Bacillus thuringiensis-maize hybrid in the sugarcane borer (Lepidoptera: Crambidae)

A novel F2 screening technique was developed for detecting resistance in sugarcane borer, Diatraea saccharalis (F.), to transgenic Bacillus thuringiensis (Bt)-maize expressing the Cry1Ab insecticidal protein. The F2 screening method involved (i) collecting larvae from maize fields; (ii) establishing two-parent families; (iii) screening F2 neonates for survival on Bt-maize leaf tissues; and (iv) confirming resistance on commercial Bt-maize plants. With the F2 screening method, 213 iso-line families of D. saccharalis were established from field collections in northeast Louisiana, USA and were screened for Bt resistance. One family was confirmed to carry a major Bt resistance allele(s). In a laboratory bioassay, larval mortality of the Bt-resistant D. saccharalis on Bt-maize leaf tissues was significantly lower than that of a Bt-susceptible strain. This Bt-resistant D. saccharalis population is the first corn stalk borer species that has completed larval development on commercial Bt-maize. The F2 screening protocol developed in this study could be modified for detecting Bt resistance alleles in other similar corn stalk borers, such as the European corn borer, Ostrinia nubilalis (Hübner), and the southwestern corn borer, D. grandiosella Dyar.     

Corn borer (Lepidoptera: Noctuidae and Crambidae) resistance of main races of maize from North America

Resistance to corn borers, Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae) and Sesamia nonagrioides (Lefebvre) (Lepidoptera: Noctuidae) in maize, Zea mays L., populations is partial, and more resistant populations are needed. The objective of this research was to compare resistance to corn borers of the main maize races from North America. Twenty open-pollinated maize populations belonging to the races Southern Dent, Corn Belt Dent, and Northern Flint, and three check populations, were evaluated under artificial infestation with S. nonagrioides and O. nubilalis. None of the populations had complete resistance. Northern Flint had the lowest yield under corn borer infestation, whereas Southern Dent had the highest yield but also the largest damage. Corn Belt Dent had a shorter growing cycle and similar yield of infected plants than Southern Dent. The checks had intermediate yield and resistance and were not significantly different from Corn Belt Dent for any trait. The Southern Dent populations Tennessee Red Cob and White Dent (PI221885 and PI311232) could be used as sources of tolerance to corn borers, although they are not expected to provide great gains compared with the levels of tolerance already present in some Corn Belt Dent and European Flint populations and would require adaptation to short growing cycle. The Corn Belt Dent synthetic BS17 had the highest yield and general agronomic performance under corn borer infestation, along with Rustler and Silver King, and the European Flint composite EPS13.     

Evaluation of oviposition deterrence in management of resistance to transgenic corn by European corn borer (Lepidoptera: Crambidae)

We modified an existing model for European corn borer, Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae), population dynamics and genetics to evaluate the effectiveness of oviposition deterrence in transgenic fields for resistance management. We simulated two types of deterrence: one type has females reducing their oviposition because of lost opportunities to lay eggs (eggs lost), and the other type has the deterred females moving to the refuge to lay eggs. Oviposition deterrence was clearly effective in extending the time to resistance to transgenic corn (R allele) in the European corn borer, particularly when 80% or more of the eggs were deterred from being oviposited on the transgenic plants. With 90% of eggs deterred, the time required to reach 50% R-allele frequency increases 3.7- to 5.5-fold compared with the no-deterrence scenario. The time to 50% R-allele frequency was similar for the two types of simulated deterrence, but the densities of the European corn borer were 100-fold higher when the deterred females oviposited in the refuge. The Y allele for insensitivity or resistance to deterrence never reached 50% within the 50-yr time line for these simulations except when the R allele was dominant and the Y allele was not recessive. The time to 50% Y-allele frequency was 33 and 26 yr when the Y allele was additive or dominant, respectively, when 50% of the eggs were deterred, but the time decreased to 18 and 16 yr when 90% of the eggs were deterred. The effectiveness of oviposition deterrence on time to resistance to transgenic insecticidal plants was not changed much when we altered our assumptions about behavior in a sensitivity analysis.     

Transgenic Bacillus thuringiensis corn hybrid performance against univoltine ecotype European corn borer (Lepidoptera: Crambidae) in South Dakota

The performances of Bt-corn hybrids against univoltine ecotype European corn borer larvae were evaluated in South Dakota from 1997 to 1999. The corn hybrids were exposed to natural seasonal fluctuations of a univoltine ecotype European corn borer population. Larval injury parameters, grain yields, and gross incomes were quantified during the 3-yr study. In general, the Bt-corn hybrids had significantly higher yields than the untreated non-Bt isolines in 1997 and 1998 when corn borer pressures were high. However, most of the Bt-corn hybrids did not produce significant yield advantages in 1999 when the European corn borer pressure was low. Some of the Bt-corn hybrids even produced significantly lower yields than their untreated non-Bt counterparts in 1999. The performances of non-Bt isolines that were treated with permethrin granules at whorl stage were similar to Bt-corn hybrids in 1998 when the corn borer pressure was high and similar to the untreated non-Bt isolines in 1999 when the corn borer pressure was low. Injury-free corn produced by Bt-corn hybrids did not necessarily translate into higher yields in some hybrid groups. Grain moisture at harvest, which can result in moisture penalty or dockage, was significantly higher in most Bt-corn hybrids. Gross incomes of the Bt-corn hybrids were generally higher than the untreated non-Bt isolines when the corn borer infestation was high, but were either similar to or lower than the untreated non-Bt isolines when the corn borer infestation was low.     

Inheritance of resistance to the Cry1Ab Bacillus thuringiensis toxin in Ostrinia nubilalis (Lepidoptera: Crambidae)

Laboratory selection with Cry1Ab, the predominant Bacillus thuringiensis (Bt) toxin in transgenic corn, Zea mays L., produced >1000-fold resistance in two laboratory strains of European corn borer, Ostrinia nubilalis (Hübner). We tested the offspring of various crosses to determine the mode of inheritance of resistance to Cry1Ab. Patterns of inheritance of resistance were similar in the two resistant strains. The progeny of reciprocal F1 crosses (resistant male x susceptible female and vice versa) responded alike in bioassays, indicating autosomal inheritance. The median lethal concentrations (LC50 values) of F1 were intermediate between the resistant and susceptible parents, indicating approximately additive inheritance. However, the dominance of resistance increased as the concentration of Cry1Ab decreased. Analysis of progeny from backcrosses (F1 x susceptible strain) suggests that resistance was controlled by more than one locus. In particular, the fit of observed to expected mortality improved as the number of putative loci increased from 1 to 10. The polygenic nature of resistance in these two laboratory strains suggests that major genes for resistance to Cry1Ab were not common in the founding populations of O. nubilalis. A low initial frequency of major genes for Cry1Ab resistance might be an important factor in delaying evolution of resistance to Bt corn in this pest.     

Effects of Bacillus thuringiensis toxin Cry1Ac and Beauveria bassiana on Asiatic corn borer (Lepidoptera: Crambidae)

In this study, interactions between Cry1Ac, a toxic crystal protein produced by Bacillus thuringiensis (Berliner), and Beauveria bassiana on the mortality and survival of Ostrinia furnacalis was evaluated in the laboratory. The results showed that Cry1Ac is toxic to O. furnacalis. Not only were larval growth and development delayed, but pupation, pupal weight and adult emergency also decreased when larvae were fed on artificial diet containing purified Cry1Ac toxin. When third instars O. furnacalis were exposed to combination of B. bassiana (1.8 × 10⁵, 1.8 × 10⁶ or 1.8 × 10⁷ conidia ml⁻¹) and Cry1Ac, (0.2 or 0.8 μg g⁻¹), the effect on mortality was additive, however, the combinations of sublethal concentrations showed antagonism between Cry1Ac (3.2 or 13 μg g⁻¹) and B. bassiana (1.8 × 10⁵ or 1.8 × 10⁶ conidia ml⁻¹). When neonates were reared on sublethal concentrations of Cry1AC until the third instar, and survivors exposed B. bassiana conidial suspension, such treatments showed additive effect on mortality of O. furnacalis except for the combination of Cry1Ac (0.2 μg g⁻¹) and B. bassiana (1.8 × 10⁶ conidia ml⁻¹) that showed antagonism.     

Southwestern corn borer (Lepidoptera: Crambidae) damage and aflatoxin accumulation in maize

Aflatoxin, a potent carcinogen, is produced by the fungus Aspergillus flavus Link: Fr. Drought, high temperatures, and insect damage contribute to increased levels of aflatoxin contamination in corn, Zea mays L. Plant resistance is widely considered a desirable method of reducing aflatoxin contamination. Germplasm lines with aflatoxin resistance have been developed. This investigation was undertaken to determine whether crosses among these lines exhibited resistance to southwestern corn borer, Diatraea grandiosella Dyar, and to assess the effects of southwestern corn borer feeding on aflatoxin accumulation. Differences in ear damage among southwestern corn borer infested hybrids were significant. Estimates of general combining ability effects indicated that the lines Mp80:04, Mp420, and Mp488 contributed to reduced ear damage, and SC213 and T165 contributed to greater damage when used in hybrids. Mean aflatoxin levels were 254 ng/g for hybrids infested with southwestern corn borer larvae and 164 ng/g for noninfested hybrids in 2000 when environmental conditions were conducive to aflatoxin production. In contrast, the overall mean aflatoxin level for southwestern corn borer infested hybrids was only 5 ng/g in 1999 when environmental conditions did not favor aflatoxin accumulation. Crosses that included lines selected for aflatoxin resistance as parents (Mp80:04 and Mp313E) exhibited lower levels of aflatoxin contamination both with and without southwestern corn borer infestation in 2000. Only the experimental line Mp80:04 contributed significantly to both reduced southwestern corn borer damage and reduced aflatoxin contamination.     

Frequency of resistance to Bacillus thuringiensis toxin Cry1Ab in southern United States Corn Belt population of European corn borer (Lepidoptera: Crambidae)

The high-dose refuge resistance management strategy is the main approach used to delay resistance in targeted pests to Bacillus thuringiensis (Bt) toxins in transgenic crops. We used an F2 screen to test a critical assumption of the high-dose refuge strategy, which is that resistance allele (R) frequencies are initially rare (<10(-3)) in Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae) from the southern Corn Belt. We expanded the methodological scope of the F2 screen so that both males and females may be used to initiate a screen and determined how the results from both sexes may be combined. In total, 62 female and 131 male O. nubilalis lines from Kansas and 39 female and four male lines from Texas were screened. No major resistance alleles were found and estimated R frequency for the southern Corn Belt was updated to between 0 and 0.0044 with 95% credibility. The experiment-wise detection probability was 98.7%. These results suggest the frequency of resistance alleles is low enough that the high-dose refuge resistance management strategy may be effective for delaying resistance evolution in O. nubilalis to Bt corn in the southern Corn Belt.