Transgenic Bt corn varietal resistance against the Mexican rice borer,Eoreuma loftini (Dyar) (Lepidoptera: Crambidae) and implications to sugarcane

The Mexican rice borer, Eoreuma loftini (Dyar), attacks crops including corn, Zea mays L., rice, Oryza sativa L., sorghum, Sorghum bicolor (L.) Moench, and sugarcane, Saccharum spp. Strongly resistant varieties of any kind, native or otherwise, have not been identified. A field plot corn variety test using two transgenic Bt varieties, Pioneer 31G71, expressing the Cry1F insecticidal protein, and Golden Acres 28V81, expressing the Cry1A.105, Cry2Ab2, and Cry3Bb1 insecticidal proteins, and two non-Bt controls, Dekalb DKC 69-72 and BH Genetics 9050, all four commonly grown in the Lower Rio Grande Valley of Texas, showed that, although oviposition preference was not affected, 28V81 resisted larval stalk boring to the extent that Mexican rice borer injury was almost non-existent. Pioneer 31G71 was infested nearly as much as the controls, but larval development to adulthood was reduced by ≈70%. Rearing larvae on 5, 50, 500, and 5000 μg of corn leaf tissue per ml of artificial diet showed that, while the three lowest concentrations did not affect larval growth and development, the high concentration of 28V81 reduced survivorship to the pupal stage, decreased weight of 4-wk-old larvae, and prolonged development to pupation. Lower numbers of pheromone trap-captured adults at the edges of commercial Bt and non-Bt corn fields showed that populations were lower at the Bt cornfields, suggesting a lesser rate of adult production. Because corn is a preferred host plant over sugarcane, sorghum or rice, use of resistant transgenic Bt corn varieties will likely protect the crop from the substantial injury that can be caused by the pest. This study also suggests that Bt genes might result in similarly strong resistance when inserted in other vulnerable crops such as sugarcane.     

Theoretical and empirical assessment of a seed mix refuge in corn for southwestern corn borer

Larval movement in seed mixes of Bt and non-Bt corn (Zea mays L.) can potentially increase the rate at which resistance evolves compared to a structured refuge. Pyramided Bt crops with multiple efficacious Bt toxins make it less likely that interplant larval movement will be successful within a seed mix field but high levels of larval movement may decrease refuge value as larvae moving from non-Bt plants die on neighboring Bt plants. Estimates of refuge productivity obtained from multi-site field studies evaluating larval movement and survival of the southwestern corn borer (SWCB), Diatraea grandiosella Dyar, in MON 89034 seed mix refuge fields were compared to predictions from model simulations of larval movement within a seed mix refuge. Results from field studies and modeling simulations of interplant movement show that seed mix refuge was a productive source of susceptible SWCB and that successful interplant movement by larvae within a seed mix refuge was unlikely to occur at frequency that would increase the fitness of resistant heterozygotes or low level resistance mechanisms. Combined, these results indicate that MON 89034 with a 5% seed mix refuge is a durable IRM tactic for SWCB across a range of larval movement and refuge assumptions and show that a 5% seed mix refuge can delay resistance longer than a similar-sized structured refuge given the nature of grower compliance with refuge requirements.     

Aflatoxin contamination of corn under different agro-environmental conditions and biocontrol applications

Biological control of the fungus Aspergillus flavus has been shown to be effective in reducing aflatoxin contamination in corn. This study compared field application of a bioplastic-based formulation for delivering atoxigenic A. flavus isolates in Northern Italy and the Mississippi Delta.Due to an extremely hot and dry summer at the Italy site in 2012, aflatoxin contamination was approximately seven times higher than in 2011. In 2011, and 2012, application of bioplastic granules inoculated with the atoxigenic isolate A. flavus NRRL 30797 at 15 and 30 kg ha⁻¹ resulted in a reduction of aflatoxin contamination by 67.2 ± 4.1% and 94.8 ± 5.3%, respectively. The higher application rate was also effective when soil abundance of A. flavus was artificially increased by applying contaminated corn residues. At the Mississippi site, summer 2012 was also hot and dry, with high levels of aflatoxin contamination. In fields planted with non-Bt or Bt hybrids, application of biocontrol granules inoculated with A. flavus NRRL 30797 or NRRL 21882 at 30 kg ha⁻¹ reduced aflatoxin contamination to up to 89.6%. Field experiments on two continents showed that bioplastic-based A. flavus formulations markedly reduced aflatoxin contamination under different agro-environmental conditions and infestation intensities.     

Resistance of Bt-maize (MON810) against the stem borers Busseola fusca (Fuller) and Chilo partellus (Swinhoe) and its yield performance in Kenya

A study was conducted to assess the performance of maize hybrids with Bt event MON810 (Bt-hybrids) against the maize stem borer Busseola fusca (Fuller) in a biosafety greenhouse (BGH) and against the spotted stem borer Chilo partellus (Swinhoe) under confined field trials (CFT) in Kenya for three seasons during 2013–2014. The study comprised 14 non-commercialized hybrids (seven pairs of near-isogenic Bt and non-Bt hybrids) and four non-Bt commercial hybrids. Each plant was artificially infested twice with 10 first instar larvae. In CFT, plants were infested with C. partellus 14 and 24 days after planting; in BGH, plants were infested with B. fusca 21 and 31 days after planting. In CFT, the seven Bt hybrids significantly differed from their non-Bt counterparts for leaf damage, number of exit holes, percent tunnel length, and grain yield. When averaged over three seasons, Bt-hybrids gave the highest grain yield (9.7 t ha⁻¹), followed by non-Bt hybrids (6.9 t ha⁻¹) and commercial checks (6 t ha⁻¹). Bt-hybrids had the least number of exit holes and percent tunnel length in all the seasons as compared to the non-Bt hybrids and commercial checks. In BGH trials, Bt-hybrids consistently suffered less leaf damage than their non-Bt near isolines. The study demonstrated that MON810 was effective in controlling B. fusca and C. partellus. Bt-maize, therefore, has great potential to reduce the risk of maize grain losses in Africa due to stem borers, and will enable the smallholder farmers to produce high-quality grain with increased yield, reduced insecticide inputs, and improved food security.     

Evaluation of European corn borer Ostrinia nubilalis (Lepidoptera: Crambidae) larval movement and survival in structured and seed blend refuge plantings

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 (Hübner), 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.     

Screening,identification and antagonistic effect of antagonistic bacteria JTFM1001 against aflatoxin contamination in corn

Aflatoxin is a strong carcinogenic and toxic fungal toxin produced by Aspergillus flavus and other Aspergillus species,and can seriously threaten the health of consumers,thus becoming a global concern.Corn,as an important oil and economic crop,is highly susceptible contaminated by aflatoxin.In this study,antagonistic bacteria with strong inhibitory effect on aflatoxin were screened to provide support for the treatment aflatoxin contamination control in corn.Ten strains which have strong antagonistic effects against A.flavus were isolated from healthy corn from different corn producing areas in China.Among them,the antagonistic bacteria JTFM1001 through corn kernels in vivo and field experiment,the inhibition effect of aflatoxin contamination reached above 70%and 55%,respectively.And the strain was identified as Bacillus subtilis based on its morphological,physiological and biochemical characteristics and phylogenetic analysis of 16S rDNA.In addition,our data showed that it can colonize in the rhizosphere and survive for a long time,forming the dominant flora,with broad application prospect.Finally,we were surprised to find that the antibacterial metabolites secreted by the antagonistic bacteria was one of the mechanisms of its inhibition of A.flavus and aflatoxin.This will provide us with new ideas and perspectives on the effective prevention and control of aflatoxin contamination in corn and corn oil.     

转Bt基因玉米的抗螟性及产量分析

在玉米心叶期和穗期对3个转Bt基因玉米品种及其相对应的受体品种进行人工接虫,分析了转Bt基因玉米对玉米螟的抗性及对产量损失的影响。结果表明：转Bt基因玉米在玉米心叶期,对一代玉米螟表现为高抗;在穗期,对二代玉米螟也表现高抗。在接虫条件下,转Bt基因玉米可以减少产量损失30%以上。因此,种植转Bt基因玉米具有明显增产作用和经济效益。     

Interaction between host plant resistance and biological activity of Bacillus thuringiensis in managing the pod borer Helicoverpa armigera in chickpea

The legume pod borer Helicoverpa armigera (Hubner) (Lepidoptera: Noctuidae) has developed high levels of resistance to conventional insecticides, and therefore, efforts are being made to develop transgenic chickpea expressing toxin genes from the bacterium Bacillus thuringiensis (Bt) for controlling this pest. However, there is an apprehension that acid exudates in chickpea might interfere with the biological activity of Bt. Therefore, we studied the biological activity of Bt (Biolep^R) on four chickpea genotypes with different levels of resistance to H. armigera under field conditions, and by incorporating lyophilized leaf and pod tissue into the artificial diet with and without Bt. The pH of the acid exudates varied from 2.1 to 2.9, and malic and oxalic acids were the major components of the acid exudates in different chickpea genotypes. There was no survival of H. armigera larvae in chickpea plants sprayed with 0.1, 0.2 and 0.5% Bt. There was a significant reduction in larval survival, larval and pupal weights and fecundity, and prolongation of larval and pupal periods in chickpea plots sprayed with Bt (0.05%) as compared to the unsprayed plots. Biological activity of Bt was lower on artificial diets with leaf or pod powder of chickpea genotypes, which might be because of a low intake of Bt toxins due to the antifeedant effects of acid exudates in the chickpea or reduction in biological activity of Bt due to the interaction of biochemical constituents in chickpea with the Bt toxins. Larval survival, larval and pupal weights, pupation and adult emergence were significantly lower on diets with leaf or pod powder of the H. armigera-resistant genotypes than on the susceptible check. Chickpea genotypes with resistance to H. armigera acted in concert with Bt to cause adverse effects on the survival and development of this insect. The results suggested that development of transgenic chickpeas expressing toxin genes form Bt will be quite effective for controlling of the pod borer, H. armigera.     

Breeding and Characterization of a New Rice Restorer Line Containing Bt Gene

Bt5198, a new rice restorer line containing Bt gene, was developed from the cross and backcross of the elite restorer line Chenghui 177 with Bt Minghui 63, a transgenic Bt restorer line. The inbred lines were evaluated using PCR amplification, test paper evaluation, insect resistance evaluation in both the laboratory and paddy fields, nursery evaluation of rice blast resistance and pedigree selection of agronomic traits. Larval mortalities on Bt5198 and Bt Minghui 63 were 100% when rice culms were inoculated with the eggs of the striped stem borer (SSB) in the laboratory. Bt5198 was highly resistant against SSB and the yellow stem borer (YSB) under field conditions. The F₁ hybrids derived from Bt5198 and four cytoplasmic male sterile (CMS) lines were also highly resistant to SSB and YSB and had a significant heterosis. Two-year evaluation of rice blast resistance confirmed that the resistance levels of Bt5198 to leaf blast and neck blast were similar to those of Chenghui 177 and significantly better than those of Bt Minghui 63. Seed germination ability and pollen yield of Bt5198 were similar with Chenghui 177, suggesting that the introduction of the Bt gene into the new restorer line had no significant effects on seed vitality or the yield of seed production. To identify the presence of the Bt gene, it was effective to combine test paper examination with the evaluation of insect-resistance, both in the laboratory and under field conditions.     

Diallel analysis of aflatoxin accumulation in maize

Since its discovery in numerous feedstuffs, aflatoxin, a carcinogenic compound produced by the fungus Aspergillus flavus Link ex Fries, has caused much concern among consumers and producers alike. This toxin poses a serious economic threat to maize (Zea mays L.) producers of the southeastern and midwestern regions of the United States. Efforts to identify maize germplasm that is resistant to aflatoxin accumulation and to investigate the genetic basis of this resistance have been undertaken at numerous research institutions. The objectives of this study were to (1) evaluate aflatoxin accumulation in grain harvested from maize inbred lines and a diallel cross among these lines, (2) determine the importance of general and specific combining abilities in inheritance of resistance to aflatoxin accumulation, and (3) estimate general and specific combining ability effects associated with resistance to aflatoxin accumulation in the inbred lines and crosses among them. Eight inbred lines and a diallel cross of the maize lines were inoculated with an A. flavus spore suspension 12–14 d after silk emergence. Following harvest, aflatoxin content was determined from samples of grain. Statistical analyses performed using SAS general linear models (GLM) and DIALLEL-SAS indicated that general and specific combining ability were significant sources of variation in the inheritance of resistance to aflatoxin accumulation. The inbred line Mp313E, which was developed and released as a source of resistance to aflatoxin contamination, had significantly lower aflatoxin accumulation than other lines. Mo18W exhibited excellent general combining ability for reduced aflatoxin accumulation when crossed with the other lines. Both Mp313E and Mo18W could be useful in breeding programs to develop aflatoxin-resistant maize hybrids. Mp339, SC212M, and Ab24E demonstrated aflatoxin susceptibility as both inbreds and in single crosses.     

Larval survival and plant injury of Cry1F-susceptible, -resistant,and -heterozygous fall armyworm (Lepidoptera: Noctuidae) on non-Bt and Bt corn containing single or pyramided genes

The fall armyworm, Spodoptera frugiperda (J.E. Smith), is a major target of transgenic corn, Zea mays L., expressing Bacillus thuringiensis (Bt) proteins in both North and South America. A highly Cry1F-resistant strain of S. frugiperda was established from a field collection in Puerto Rico in 2011. In this study, three greenhouse trials were conducted to evaluate larval survival and leaf injury of Cry1F-susceptible, -resistant, and -heterozygous genotypes of S. frugiperda on whole plants of five non-Bt and eight Bt corn hybrids. The Bt corn products included two single-gene Bt corn hybrids containing Herculex^®I (Cry1F) and YieldGard^® (Cry1Ab) traits and six pyramided Bt corn hybrids representing four traits: Genuity^® VT Double Pro™, Genuity^®VT Triple Pro™, Genuity^® SmartStax™, and Agrisure^® Viptera™ 3111. In each trial, neonates of S. frugiperda were placed into the plant whorls at vegetative plant stages (V6–V10). Larvae of the three insect genotypes on non-Bt corn hybrids survived well and caused serious plant injury. Cry1Ab corn was ineffective against all three insect genotypes. On Cry1F corn plants, resistant larvae survived on 72.9% plants after 12–15 d and caused a leaf injury rating (Davis' 1 to 9 scales) of 5.7 after 7 d and 7.6 after 12–15 d. Both the larval survivorship and leaf injury rates of the resistant larvae on Cry1F corn plants were not significantly different from those observed on non-Bt corn hybrids. In contrast, no live larvae and little or no leaf injury were observed on the Cry1F corn plants that were infested with susceptible or heterozygous genotypes, or on the pyramided Bt plants. The results demonstrated that the Cry1F-resistant S. frugiperda was highly resistant to whole plants of Cry1F corn and the resistance was recessive. Hybrids that contained one of the four pyramided Bt traits were effective for managing the Cry1F resistance in S. frugiperda.     

Ear rot,aflatoxin accumulation,and fungal biomass in maize after inoculation with Aspergillus flavus

Aflatoxin, a toxin produced by the fungus Aspergillus flavus Link: Fries, occurs naturally in maize (Zea mays L.). Aflatoxin is a potent human carcinogen and is also toxic to livestock, pets, and wildlife. When contaminated with aflatoxin, the value of maize grain is markedly reduced. This investigation was conducted to compare ear rot, aflatoxin accumulation, and fungal biomass in maize single crosses with varying degrees of resistance to aflatoxin accumulation and to determine the relative importance of general combining ability (GCA) and specific combining ability (SCA) in the inheritance of resistance to ear rot, aflatoxin accumulation, and fungal biomass. Eight germplasm lines with different levels of resistance to aflatoxin accumulation were used as parents of a diallel cross. The cross was evaluated for visible ear rot, aflatoxin accumulation, and A. flavus infection in the grain. A. flavus infection was determined by quantitative real-time polymerase chain reaction (qRT-PCR) assays. Both GCA and SCA were significant sources of variation in the inheritance of the three traits although GCA accounted for a greater portion of the variation among single crosses. The interactions of GCA and SCA with years were highly significant for aflatoxin accumulation, but not significant for A. flavus infection. Estimates of GCA effects were highly significant for both reduced A. flavus infection and reduced aflatoxin accumulation for Mp313E, Mp715, and Mp717. Conversely, GCA effects associated with GA209 were significant for reduced levels of A. flavus infection and ear rot, but high levels of aflatoxin accumulation. Mp313E, Mp715, and Mp717 should be useful in breeding programs targeting both reduced levels of fungal infection and aflatoxin accumulation.     

Alternate crop and weed host plant oviposition preferences by the Mexican rice borer (Lepidoptera: Crambidae)

The Mexican rice borer, Eoreuma loftini (Dyar), is the key pest of sugarcane, Saccharum hybrids, in south Texas, having largely displaced the sugarcane borer, Diatraea saccharalis (F.), and it is moving into rice- and sugarcane-growing areas of east Texas and Louisiana. While a number of alternative weed and crop hosts have been reported, the extent to which they might support Mexican rice borer populations is unknown. This study involved choice assays that compared oviposition preference for and larval infestations of five mature graminaceous weed species. Levels of infestation between sugarcane and corn, Zea mays L., crop hosts and between corn and sorghum, Sorghum bicolor (L.) Moench, were also assessed. We determined that the average number of larval entry holes in sudangrass stems was ≥2.5-fold more than for any of the other four weed host plants, that corn had ≥5.9-fold more larval entry holes than sorghum and ≥8.2-fold more than sugarcane. Greater oviposition and infestation of one non-crop host over another was not related to numbers of stems per plant, but was associated with the greater stem diameter and abundance of dry leaf tissue found in Sudangrass, Sorghum bicolor (L.) Moench ssp. drummondi (Nees ex Steud.) de Wet & Harlan, johnsongrass, S. halepense (L.) and barnyardgrass, Echinochloa crus-galli (L.) P. Beauv.; relative to the other weed species in this study. In terms of the crop plants, stalk diameter and quantity of dry leaf tissue were not associated with numbers of eggs or larval entry holes in the choice assays between corn and sorghum, and between sugarcane and corn. While corn has been known as a host of the Mexican rice borer for at least 84 yr, its role in area-wide population dynamics and control efforts has likely been greatly underestimated.     

转Bt基因抗虫玉米发展状况及对非靶标昆虫的影响

转Bt基因抗虫玉米是目前商品化进程最快的转基因作物之一,并被广泛应用于农业生产,其抗虫性和经济效益已得到普遍肯定,但外源基因的导入一定程度上改变了玉米自身的基因序列,因此转Bt基因抗虫玉米对生态安全的影响也备受关注。本文从转Bt基因抗虫玉米发展现状及对非靶标昆虫影响的角度对转Bt基因玉米的安全性研究加以综述。     

Efficacy of SmartStax® insect-protected corn hybrids against corn rootworm: The value of pyramiding the Cry3Bb1 and Cry34/35Ab1 proteins

Monsanto Company and Dow AgroSciences have used conventional breeding techniques to develop the combined-trait corn (Zea mays L.) product MON 89034 × TC1507 × MON 88017 × DAS-59122-7 (“SmartStax^® corn”) that confers insect resistance against key lepidopteran pests and the corn rootworm complex (Diabrotica spp.), as well as herbicide tolerance. This product contains the Cry3Bb1, Cry34Ab1, and Cry35Ab1 proteins for corn rootworm (CRW) control. Replicated field trials were conducted in 2006 and 2007 to test the efficacy of Cry3Bb1, Cry34Ab1/Cry35Ab1, and SmartStax under both natural and controlled infestations. In both years, root damage and adult beetle emergence were significantly less for Bt-containing hybrids than for non-Bt controls. In trials with heavy insect pressure, adult beetle emergence (a measure of larval control) was significantly reduced for SmartStax hybrids compared with hybrids with the individual traits and with non-Bt controls. Similarly, strip-plot testing in 2012 in grower fields with high levels of root injury to non-Bt hybrids showed significantly lower feeding on SmartStax than on the non-Bt control or on either single-trait product. In grower fields where single-trait Cry3Bb1 products incurred heavy CRW damage in 2011, SmartStax provided consistent protection against CRW in 2012. The combination of these insecticidal proteins in a single plant provides better rootworm control than current single-trait Bt corn products and represents an effective approach for corn rootworm resistance management.     

The use of neutron tomography for the structural analysis of corn kernels

Neutron tomography was studied as a technique for non-destructively analyzing the internal structure of dried corn kernels. The study had two goals: first, to determine if the analysis could identify well-known anatomical features of the kernels; and second, to determine if it could distinguish between different types and treatments of kernels. Specifically, kernels which were infected vs. uninfected with the aflatoxin-producing fungus Aspergillus flavus were analyzed. Two different varieties of corn were used: VA35 (susceptible to A. flavus infection) and GT-MAS:gk (resistant). It was found that many anatomical features of the kernels could be identified using neutron tomography, including the scutellum, endosperm, aleurone, pericarp, pedicel, coleorhizae, radical, plumule, and coleoptile. Furthermore, differences were detected between susceptible kernels that had been inoculated and those that had not. Infected kernels were found to have lower neutron attenuation in the scutellum and embryo regions, possibly caused by lower hydrogen concentrations due to fungal degradation. No systematic structural differences were detected between resistant inoculated and resistant uninoculated kernels, as expected. This study indicated that neutron tomography could be a useful technique for the structural analysis of corn, and possibly other grains or small biological objects.     

转基因玉米SW12-859的抗螟性及农艺性状评价

SW12-859为吉林省农业科学院农业生物技术研究所培育的转基因玉米杂交组合,全株表达Cry1F基因杀虫蛋白。本研究以其为材料,与其遗传背景相似的郑单958为非转基因对照,在玉米心叶期(6~8叶期)和穗期(13~16叶期)进行田间人工接玉米螟初孵幼虫,调查其抗螟性、农艺性状及对产量损失的影响。结果表明,在玉米心叶期和穗期,转基因玉米SW12-859对一、二代玉米螟表现为高抗,郑单958表现为感,二者受玉米螟危害程度(抗螟性)差异显著。转基因玉米SW12-859和郑单958的株高、穗位高等农艺性状相近,在接虫条件下,转基因玉米产量显著高于非转基因玉米。     

Movement and survival of corn rootworm in seed mixtures of SmartStax® insect-protected corn

SmartStax^® insect-protected corn (Zea mays L.) contains genes for six Bacillus thuringiensis (Bt) proteins controlling both lepidopteran pests and the corn rootworm complex (Diabrotica spp.). The properties of SmartStax, particularly the multiple effective modes of action (i.e., each Bt protein provides a high level of control of the target pests with a low probability of cross-resistance among the proteins), have provided the opportunity to add to previously approved structured refuge options by combining the non-Bt refuge seed with SmartStax seed in a seed mix. Seed mixes ensure that a refuge is present in every Bt field, remove concerns about grower compliance with refuge requirements, and provide grower convenience. However, seed mixes could increase the likelihood that larval insects move between Bt and non-Bt plants and vice versa. Assessing the insect resistance management (IRM) value of a seed mix refuge requires an assessment of the amount of larval movement, and the consequences of that movement, for the key target pests. The studies here present such data for control of corn rootworm by SmartStax corn, which contains the rootworm-active protein Cry3Bb1 and the binary protein Cry34Ab1/Cry35Ab1. In a growth chamber experiment, SmartStax was most effective against first instars and significantly effective against second instars, but did not control third instars. In a field study of movement from a heavily infested non-Bt plant onto surrounding plants, a larger percentage of insects successfully dispersed from the infested plant when the surrounding plants were non-Bt plants than when they were SmartStax plants. A paired-plant study showed that few larvae migrated from infested SmartStax plants and survived on nearby non-Bt plants; larvae that migrated from infested non-Bt plants had low survival if the adjacent plants were SmartStax. Replicated field studies of plant-to-plant movement indicated that the non-Bt plants in a 5% or 10% seed mix consistently supported large populations of susceptible insects and represented a productive refuge, whereas the SmartStax plants had few or no survivors. The timing of emergence from seed mix plots containing 5% or 10% non-Bt plants was more similar to that of the non-Bt plots than that of the SmartStax plots. Thus, the available growth chamber and field data indicate that a seed mix of 5% or more will provide an effective refuge for corn rootworm in SmartStax corn.     

Monitoring Cry1Ab susceptibility in Asian corn borer (Lepidoptera: Crambidae) on Bt corn in the Philippines

Between 2002 and 2004, collections of egg masses of Asian corn borer (ACB), Ostrinia furnacalis (Guenée) were made from corn-planting sites on the major Philippine islands of Luzon (Laguna, Pangasinan, Camarines Sur and Isabela provinces) and Mindanao (Bukidnon and South Cotabato provinces). The resulting neonates were bioassayed for susceptibility to Bacillus thuringiensis (Bt) Cry1Ab protein. The median lethal concentrations (LC₅₀s) for the different collections ranged from 0.42 to 2.37 ng/cm². The bioassay results suggest that Philippine corn borer populations were highly susceptible to Cry1Ab protein prior to the widespread deployment of Bt corn. The upper limit of the estimated LC₉₉ (104 ng/cm²) from the pooled bioassay data was selected as a candidate diagnostic concentration and subsequently tested on eleven ACB populations. Results of the validation assays showed that the mortality response of all the tested ACB populations was higher than the expected mortality (99%). Therefore, the concentration of 104 ng/cm² was used to monitor susceptibility in ACB populations in the Philippines. Monitoring of field populations during 2009 in areas where Bt corn had been grown for 3 years found some enhanced survival of neonates at the diagnostic concentration but progeny of the diagnostic-concentration survivors did not survive on Bt corn, indicating that ACB populations in the Philippines remain susceptible to Cry1Ab-containing Bt corn hybrids.     

Susceptibility of Cry1Ab-susceptible and -resistant sugarcane borer to transgenic corn plants containing single or pyramided Bacillus thuringiensis genes

Multiple independent trials were conducted to evaluate the performance of Cry1Ab-susceptible (Cry1Ab-SS), -heterozygous (Cry1Ab-RS), and -resistant (Cry1Ab-RR) genotypes of the sugarcane borer, Diatraea saccharalis (F.), on eight commercial hybrids and six experimental corn lines. The commercial varieties included two non-Bt and six Bt corn hybrids that expressed a single Bt protein (either Cry1Ab or Cry1F) targeting above-ground lepidopteran pests. The six experimental lines consisted of two non-Bt and four Bt corn lines, two expressing just the Cry1Ab protein and two containing the pyramided-genes Cry1A.105 and Cry2Ab2 (event MON 89034). Larval mortality on non-Bt corn leaf tissue ranged from 6 to 45% after 12 d across insect genotypes. The 12 d mortality of Cry1Ab-SS on leaf tissue of commercial Cry1Ab or Cry1F corn was 96-100%, whereas it was 80-96% for Cry1Ab-RS and 68-78% for Cry1Ab-RR. On intact plants, 39-64% of larvae survived on non-Bt corn plants after 21-25 d. Larval survivorship on intact plants of commercial Cry1Ab or Cry1F corn was 0-8.1% for Cry1Ab-SS, 1.3-34% for Cry1Ab-RS, and 19-51% for Cry1Ab-RR. Larvae of Cry1Ab-RR and -RS also caused significant plant injury to most of the commercial Bt corn hybrids, especially to the Cry1Ab corn. Cry1Ab resistance in D. saccharalis was incompletely dominant on commercial Bt corn hybrids. However, both experimental lines with pyramided genes of Cry1A.105 and Cry2Ab2 provided complete control of all three insect genotypes in both leaf tissue and intact plant tests. Results of this study suggest that MON 89034 should offer a means for Bt resistance management in D. saccharalis.