Bt Maize Plants Research Articles

Fall armyworm (Lepidoptera: Noctuidae): practical resistance of 2 Brazilian populations to Cry1A.105 + Cry2Ab and Cry1F Bt maize.

In the Americas, transgenic cropsproducing insecticidal proteins fromBacillus thuringiensis Berliner (Bt,Bacillales: Bacillaceae) have been used widely to manage fall armyworm (FAW, Spodoptera frugiperda [J.E. Smith]). As resistance to Cry1 single-gene Bt maize (Zea mays L.)rapidly evolved in some FAW populations, pyramided Bt maizehybrids producing Cry1, Cry2, or Vip3Aa proteins were introduced in the 2010s. We examined field-evolved resistance to single- and dual-protein Bt maize hybrids in 2 locations in southeastern Brazil, where plant damage by FAW larvae far exceeded the economic threshold in 2017. We collected late-instar larvae in Cry1A.105 + Cry2Ab and Cry1F maize fields and established 2 FAW populations in the laboratory. The F1 offspring reared on the foliage of Bt and non-Bt maize plants (Cry1A.105 + Cry2Ab and Cry1F) showed neonate-to-adult survival rates as high as 70% for both populations. There was no significant difference in the life-table parameters of armyworms reared on non-Bt and Bt maize foliage, indicating complete resistance to Cry1A.105 + Cry2Ab maize. Larval survival rates of reciprocal crosses of a susceptible laboratory strain and the field-collected populations indicated nonrecessive resistance to Cry1F and a recessive resistance to Cry1A.105 + Cry2Ab maize. When relaxing the selection pressure, the armyworm fitness varied on Cry1A.105 + Cry2Ab and non-Bt maize; the resistance was somewhat stable across 12 generations, without strong fitness costs, although one of the lines died confounded by a depleted-quality, artificial rearing diet. To our knowledge, this is the first report documenting the practical resistance of FAW to a pyramided Bt crop. We discuss the implications for resistance management.

Impact of Seed Blend and Structured Maize Refuge on Helicoverpa zea (Lepidoptera: Noctuidae) Potential Phenological Resistance Development Parameters in Pupae and Adults.

Helicoverpa zea, an economic pest in the southeastern U.S., has evolved practical resistance to Bt Cry toxins in maize and cotton. Insect resistance management (IRM) programs have historically required planting of structured non-Bt maize, but because of its low adoption, the use of seed blends has been considered. To generate knowledge on target pest biology and ecology to help improve IRM strategies, nine field trials were conducted in 2019 and 2020 in Florida, Georgia, North Carolina, and South Carolina to evaluate the impact of Bt (Cry1Ab + Cry1F or Cry1Ab + Cry1F + Vip3A) and non-Bt maize plants in blended and structured refuge treatments on H. zea pupal survival, weight, soil pupation depth, adult flight parameters, and adult time to eclosion. From a very large sample size and geography, we found a significant difference in pupal mortality and weight among treatments in seed blends with Vip3A, implying that cross-pollination occurred between Bt and non-Bt maize ears. There was no treatment effect for pupation depth, adult flight distance, and eclosion time. Results of this study demonstrate the potential impact of different refuge strategies on phenological development and survival of an important pest species of regulatory concern. This article is protected by copyright. All rights reserved.

Assessing the effects of Bt maize on the non-target pest Rhopalosiphum maidis by demographic and life-history measurement endpoints.

The most commercialized Bt maize plants in Europe were transformed with genes which express a truncated form of the insecticidal delta-endotoxin (Cry1Ab) from the soil bacterium Bacillus thuringiensis (Bt) specifically against Lepidoptera. Studies on the effect of transgenic maize on non-target arthropods have mainly converged on beneficial insects. However, considering the worldwide extensive cultivation of Bt maize, an increased availability of information on their possible impact on non-target pests is also required. In this study, the impact of Bt-maize on the non-target corn leaf aphid, Rhopalosiphum maidis, was examined by comparing biological traits and demographic parameters of two generations of aphids reared on transgenic maize with those on untransformed near-isogenic plants. Furthermore, free and bound phenolics content on transgenic and near-isogenic plants were measured. Here we show an increased performance of the second generation of R. maidis on Bt-maize that could be attributable to indirect effects, such as the reduction of defense against pests due to unintended changes in plant characteristics caused by the insertion of the transgene. Indeed, the comparison of Bt-maize with its corresponding near-isogenic line strongly suggests that the transformation could have induced adverse effects on the biosynthesis and accumulation of free phenolic compounds. In conclusion, even though there is adequate evidence that aphids performed better on Bt-maize than on non-Bt plants, aphid economic damage has not been reported in commercial Bt corn fields in comparison to non-Bt corn fields. Nevertheless, Bt-maize plants can be more easily exploited by R. maidis, possibly due to a lower level of secondary metabolites present in their leaves. The recognition of this mechanism increases our knowledge concerning how insect-resistant genetically modified plants impact on non-target arthropods communities, including tritrophic web interactions, and can help support a sustainable use of genetically modified crops.

Performance of Seed Treatments Applied on Bt and Non-Bt Maize Against Fall Armyworm (Lepidoptera: Noctuidae).

Fall armyworm (FAW), Spodoptera frugiperda (J. E. Smith), is the main pest of maize in Brazil, attacking plants from emergence to reproductive stages. Here, we conducted studies to evaluate the efficacy of two seed treatments (chlorantraniliprole alone and imidacloprid combined with thiodicarb) on Bt and non-Bt maize in laboratory bioassays with distinct FAW strains that are susceptible, selected for resistance to Bt-maize single (Cry1F) or pyramided (Cry1A.105 + Cry2Ab2) events and F1 hybrids of the selected and susceptible strains (heterozygotes), and in the field against a natural infestation. In the laboratory, leaf-discs from seed treated Bt-maize plants at 7 d after emergence (DAE) increased the mortality of FAW resistant, heterozygote, and susceptible strains up to 24.8%, when compared with the respective maize grown without a seed treatment. In the field against natural infestations of FAW, Bt maize with a seed treatment had ~30% less FAW damage than non-Bt maize with the same seed treatment at 7 and 14 DAE. No differences in FAW damage was observed between Bt and non-Bt maize grown with and without a seed treatment at 21 DAE. Maize seeds treated with chlorantraniliprole alone or imidacloprid and thiodicarb combined presented limited protection against early infestations of FAW strains under laboratory and field studies.

Short communication: The rate of release of Cry1Ab protein from Bt maize leaves into water

Transgenic Bt maize plants are genetically modified to contain genes of Bacillus thuringiensis that encode for δ-endotoxins (Cry1Ab protein) that have insecticidal properties. These endotoxins target certain lepidopteran pests of maize. There are several entry routes by which Cry proteins enter the aquatic ecosystem in which aquatic organisms are exposed to these proteins. The main route is through plant debris such as leaves, stalks and postharvest detritus that are transported by means of run-off, rain and wind. While several studies have been conducted on the fate of Cry1Ab protein in terrestrial ecosystems, little is known of the release rates of Cry1Ab proteins from maize plant tissues that end up in aquatic ecosystems. In this study, leaves of Bt-maize and its isoline were submerged in containers filled with deionised or borehole water for a period of 16 days, and kept at 3 different temperatures (10±1, 21±1 and 30±1°C). Samples were collected at 1, 2, 4, 8, 16, 24, 48, 96, 192 and 384 h post submersion and analysed for Cry protein content using enzyme-linked immunosorbent assay (ELISA). The release of Cry protein from submerged maize leaves was influenced by temperature, and duration of immersion. An increase in Cry protein levels in the water was observed from the first hour onwards in both water types until the end of the experiment. The highest concentration of Cry protein was found at 30°C. This study showed that temperature and time period influence the release rate of Cry proteins from dried leaf matter into the aquatic environment.

Preference of Neoseiulus californicus (Acari: Phytoseiidae) for volatiles of Bt maize induced by multiple herbivory

Plant indirect induced defenses against herbivores are characterized by the production of plant volatiles that to attract natural enemies. The objective of this study was to evaluate whether the attack of the two-spotted spider mite Tetranychus urticae or the multiple herbivory of T. urticae together with the fall armyworm Spodoptera frugiperda are able to elicit indirect induced defense in conventional and Bt maize plants. The experiment was carried out in the laboratory using Y-tube olfactometer, evaluating the predatory mite Neoseiulus californicus olfactory preference for plant volatiles. The treatments involved: Clean Conventional Plant; Conventional Plant Infested with T. urticae; Clean Bt Plant; Bt Plant Infested with T. urticae; Conventional Plant Infested with T. urticae + fall armyworm; Bt Plant Infested with T. urticae + fall armyworm. For the chemical analyzes the Trace GC Ultra gas chromatograph was used, paired with the Polaris Q mass spectrometer, GCMS system. Neoseiulus californicus was also unable to distinguish between volatiles from both conventional and Bt infested maize plants. Moreover, there was no discrimination of the predator mite between plants under single and multiple infestations, both in conventional and Bt maize. When comparing conventional and Bt plants, both with multiple infestation, the predator mite N. californicus had no preference among these sources of odors. However, there was observed chemical changes of the volatiles among the groups of plants studied. Thus, it is suggested that the groups of plants under study have chemical modifications, but they are not able to attract N. californicus. In addition, Bt plants response was similar to conventional plants on attracting N. californicus.

Effects of Bacillus thuringiensis Genetic Engineering on Induced Volatile Organic Compounds Emission in Maize and the Attractiveness to a Parasitic Wasp.

In order to control lepidopteran and coleopteran insects, the genes expressing Bacillus thuringiensis (Bt) insecticidal proteins have been transferred into crops. Ecological risk assessments of the transgenic plants have included impacts on non-target entomophagous insects, such as parasitoid wasps. Herbivore-induced plant volatiles are considered to be important defensive traits of plants because these compounds play as an important role in recruitment of natural enemies. Here, we evaluated induced volatile emissions of maize seedlings of two Bt cultivars (5422Bt1, event Bt11 and 5422CBCL, event Mon810), and their nearly isogenic non-Bt line 5422. We damaged plants mechanically and then applied with the regurgitant of Spodoptera litura (F.) caterpillars (Lepidoptera: Noctuidae), or treated the plants with the plant hormone jasmonic acid (JA), to trigger similar defensive responses of plants. Compared to the non-Bt isoline 5422 and the Bt maize 5422CBCL, the other Bt maize 5422Bt1 released more (3E)-4,8-dimethyl-1,3,7-nonatriene (DMNT) when they were all treated by artificial wounds and caterpillar regurgitant; and released more linalool, DMNT and (E)-β-farnesene when applied with JA solution. As a result, the total volatile emission of the 5422Bt1 was highest. However, the difference in volatile emission did not affect the attractiveness of the Bt maize plants to the egg parasitoid Trichogramma ostriniae Pang et Chen (Hymenoptera: Trichogrammatidae) compared to the nearly isogenic non-Bt plants. The variability of induced volatiles of maize cultivars derived from conventional breeding programs and transgenic methods are discussed.

Field-evolved resistance of Helicoverpa zea (Boddie) to transgenic maize expressing pyramided Cry1A.105/Cry2Ab2 proteins in northeast Louisiana, the United States.

The corn earworm, Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae), is a major target pest of pyramided Bt maize and cotton in the U.S. In 2017 and 2018, notable ear damage and larval survival of H. zea were observed on pyramided Cry1A.105/Cry2Ab2 maize in some fields in northeast Louisiana, U.S. The objective of this study was to determine if the field control problem was due to resistance development to the Bt proteins in plants. A total of 15 H. zea populations were collected from Bt and non-Bt maize plants in 2017 and 2018 in multiple locations in Louisiana, Florida, and Georgia. Diet-overlay bioassays showed that LC50s of Cry1A.105 and Cry2Ab2 for populations collected from the areas with control problems in northeast Louisiana were as much as >1623- and 88-fold greater than that of a susceptible strain, respectively. In addition, two field trials in 2018 validated that Cry1A.105/Cry2Ab2 maize failed in managing natural H. zea populations, while Bt maize containing Vip3A was effective in northeast Louisiana. Results of the study documented that the observed field control problems of Cry1A.105/Cry2Ab2 maize against H. zea in northeast Louisiana were due to resistance development of the insect to the Bt proteins in plants. This is the first documentation of field-evolved resistance to pyramided Bt maize in a target insect species in southern U.S. However, susceptibility levels to Cry1A.105 and Cry2Ab2 varied greatly among populations collected from the three states, suggesting uneven distributions of the resistance in the region.

Managing fall armyworm, Spodoptera frugiperda (Lepidoptera: Noctuidae), with Bt maize and insecticides in southern Brazil.

Maize plants expressing insecticidal proteins of Bacillus thuringiensis are valuable options for managing fall armyworm (FAW), Spodoptera frugiperda, in Brazil. However, control failures were reported, and therefore insecticides have been used to control this species. Based on these, we evaluated the use of Bt maize and its integration with insecticides against FAW in southern Brazil. Early-planted Agrisure TL, Herculex, Optimum Intrasect and non-Bt maize plants were severely damaged by FAW and required up to three insecticidal sprays. In contrast, YieldGard VT Pro, YieldGard VT Pro 3, PowerCore, Agrisure Viptera and Agrisure Viptera 3 showed little damage and did not require insecticides. Late-planted Bt maize plants showed significant damage by FAW and required up to four sprays, with the exceptions of Agrisure Viptera and Agrisure Viptera 3. Exalt (first and second sprays); Lannate + Premio (first spray) and Avatar (second spray); and Karate + Match (first spray) and Ampligo (second spray) were the most effective insecticides against FAW larvae in Bt and non-Bt maize. Maize plants expressing Cry proteins exhibited FAW control failures in southern Brazil, necessitating insecticidal sprays. In contrast, Bt maize containing the Vip3Aa20 protein remained effective against FAW. However, regardless of the insecticide used against FAW surviving on Bt maize, grain yields were similar. © 2017 Society of Chemical Industry.

Quantifying rates of random mating in western corn rootworm emerging from Cry3Bb1‐expressing and refuge maize in field cages

AbstractThe western corn rootworm (WCR), Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae), is the most significant pest of field maize, Zea mays L. (Poaceae), in the USA. Maize plants expressing Bt toxins targeting the corn rootworm complex have been widely adopted and are the primary insecticidal control measure for this pest in North America. Insect resistance management tactics using various refuge structures have been adopted to ensure Bt products will retain durability. An assumption of the refuge strategy is that males and females emerging from Bt and refuge plantings mate randomly; this has not been tested in the field. We conducted cage studies using field populations of WCR in Indiana, USA, to generate empirical field data on mating rates between beetles emerging from Cry3Bb1‐expressing Bt and refuge maize plants. Two refuge configurations were tested; all refuge plants were labeled using the stable isotope 15N. This mark persists in adult beetles after eclosion, allowing for collection and analysis of isotopic ratios of all beetles. Additional data collected included adult emergence rates, timing and sex ratios for each of the treatments, and head capsule size and dry weights of beetles collected. Treatment had a significant effect on dry weight; mean dry weight decreased in Bt‐only treatments. Fisher's exact test of proportions of mating pairs of refuge and Bt insects indicated that mating was not random in 20% strip refuges and 5% seed blend treatments. We found high percentages of beetles that fed on Bt‐expressing plants as larvae, suggesting that mating between resistant beetles may not be rare even if random mating did occur.

Performance of Cry1Ab‐susceptible and ‐heterozygous resistant populations of sugarcane borer in sequential feedings on non‐Bt and Bt maize plant tissue

AbstractA seed mix strategy has been used to provide refuge to susceptible insects for resistance management in planting transgenic maize expressing Bacillus thuringiensis Berliner (Bt) proteins. To determine whether larval movement in a seed mix planting creates favorable conditions for resistant heterozygotes of a target pest, performance of Cry1Ab‐susceptible (SS) and ‐heterozygous resistant (RS) populations of the sugarcane borer, Diatraea saccharalis (Fabricius) (Lepidoptera: Crambidae), was evaluated in sequential feeding on non‐Bt and Cry1Ab (event MON 810) maize plant tissue. For each insect population, nine feeding sequences were employed. SS and RS feeding on non‐Bt plants for their entire larval stages survived well and >60% of the adult pairs produced viable eggs, with an average of 269 progeny per female, whereas none of the two populations on Bt maize plants survived to the pupal stage. SS larvae could not develop to adults if the larvae fed on non‐Bt plants for ≤15 days and then moved to Bt plants. In contrast, 4.2–29.2% of RS larvae that fed on non‐Bt plants for ≥9 days and then moved to Bt plants developed to adults, and 63.6% of pairs of these adults produced viable eggs, with an average of 185 progeny per female. For SS larvae that fed on Bt plants for 1 or 2 days and then moved to non‐Bt plants, few larvae developed to adults with varied emergence times, whereas 28.1 and 13.5% RS larvae feeding on Bt plants for 1 and 2 days, respectively, successfully developed to adults; 43.8% of pairs of these adults produced viable eggs, with an average of 220 progeny per female. For the case of the single Bt gene maize plants (event MON 810), the results suggest that RS insects may have advantages in survival and reproduction over SS if RS larvae hatch and feed on Bt plants during the first 1 or 2 days and then move to non‐Bt plants. This advantage is less for RS larvae that hatch and feed on non‐Bt plants first and then move to Bt plants, unless the larval movement occurs in the later stages (e.g., fourth or fifth instars).

Functional dominance of different aged larvae of Bt-resistant Spodoptera frugiperda (Lepidoptera: Noctuidae) on transgenic maize expressing Vip3Aa20 protein

Fall armyworm (FAW), Spodoptera frugiperda (J. E. Smith), is the main target pest of transgenic maize expressing insecticidal proteins from Bacillus thuringiensis Berliner (Bt) in Brazil. To optimize resistance management strategies, we evaluated the functional dominance of different aged larvae of Bt-resistant FAW on Vip3Aa20 maize. We measured the survival and development of Vip3Aa20-resistant, -heterozygote, and -susceptible strains on MIR162 (expressing Vip3Aa20) and Bt11 × MIR162 × GA21 (expressing Vip3Aa20 and Cry1Ab) maize. The resistant strain, from neonate to sixth instar, showed more than 72% survival on Vip3Aa20 maize. From surviving larvae, more than 64 and 54% developed to pupae and adults, respectively. In contrast, heterozygote and susceptible strains showed no larval survival up to fourth instar, and less than 25% larval survival in the fifth and sixth instar on Vip3Aa20 maize. These larvae produced less than 21% of pupae and adults. The development time of FAW strains from neonate-to-adult exposed to Vip3Aa20 maize was similar; however, the resistant strain showed an increase of ∼ 2 d when compared to those fed only non-Bt maize. In summary, the resistance of S. frugiperda to Vip3Aa20 maize is functionally recessive from neonate up to fourth instar larvae. However, high larval survival of resistant strain and some survival of heterozygote larvae in advanced instars on Vip3Aa20 maize were observed. These results will be important for designing insect resistance management to Bt maize plants expressing Vip3Aa20 protein in Brazil.

Genetic basis of Cry1F resistance in two Brazilian populations of fall armyworm, Spodoptera frugiperda

Large-scale adoption of transgenic crops expressing genes from Bacillus thuringiensis (Bt) imposes high selection pressure for evolution of field-relevant resistance that can reduce pest control efficacy, such as reported for Cry1F maize (Zea mays L.) in populations of fall armyworm, Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae), of Puerto Rico, Brazil, and the United States. As part of our effort to improve fall armyworm resistance management to Bt crops, here we determined the genetic basis of Cry1F resistance in two S. frugiperda strains originated from field collections in different regions of Brazil and further selected in the laboratory for high levels of resistance to Cry1F maize. Continuous exposure to the TC1507 event for 11 generations resulted in more than 183-fold resistance to Cry1F in the two strains studied, and such a high resistance level enabled the insects to complete larval development on the Bt maize plants. Genetic analyses using concentration-response bioassays with progenies from reciprocal crosses between resistant and susceptible insects indicated that the inheritance of the resistance is autosomal, recessive and without maternal effects. Backcross of the F1 progeny with the parental resistant strains revealed that the resistance in the two selected strains is conferred by a single locus or set of tightly linked loci. These results support some of the assumptions of the strategy in use for fall armyworm resistance management to Bt Cry1F maize, but survival rates of heterozygotes on the Bt plants were higher than 5%, showing that the Cry1F maize does not produce a high dose of the insecticidal protein for S. frugiperda. Additionally, we detected a delay in larval development time that may favor assortative mating of individuals carrying resistance alleles. These findings are consistent with the rapid evolution of Cry1F resistance in certain field populations of fall armyworm. Implications for resistance management of S. frugiperda to Bt maize are discussed.

Resistance to dual-gene Bt maize in Spodoptera frugiperda: selection, inheritance, and cross-resistance to other transgenic events.

Transgenic crop “pyramids” producing two or more Bacillus thuringiensis (Bt) toxins active against the same pest are used to delay evolution of resistance in insect pest populations. Laboratory and greenhouse experiments were performed with fall armyworm, Spodoptera frugiperda, to characterize resistance to Bt maize producing Cry1A.105 and Cry2Ab and test some assumptions of the “pyramid” resistance management strategy. Selection of a field-derived strain of S. frugiperda already resistant to Cry1F maize with Cry1A.105 + Cry2Ab maize for ten generations produced resistance that allowed the larvae to colonize and complete the life cycle on these Bt maize plants. Greenhouse experiments revealed that the resistance was completely recessive (Dx = 0), incomplete, autosomal, and without maternal effects or cross-resistance to the Vip3Aa20 toxin produced in other Bt maize events. This profile of resistance supports some of the assumptions of the pyramid strategy for resistance management. However, laboratory experiments with purified Bt toxin and plant leaf tissue showed that resistance to Cry1A.105 + Cry2Ab2 maize further increased resistance to Cry1Fa, which indicates that populations of fall armyworm have high potential for developing resistance to some currently available pyramided maize used against this pest, especially where resistance to Cry1Fa was reported in the field.

Cry1F resistance in fall armyworm Spodoptera frugiperda: single gene versus pyramided Bt maize.

Evolution of insect resistance to transgenic crops containing Bacillus thuringiensis (Bt) genes is a serious threat to the sustainability of this technology. However, field resistance related to the reduced efficacy of Bt maize has not been documented in any lepidopteran pest in the mainland U.S. after 18 years of intensive Bt maize planting. Here we report compelling evidence of field resistance in the fall armyworm, Spodoptera frugiperda (J.E. Smith), to Cry1F maize (TC 3507) in the southeastern region of the U.S. An F2 screen showed a surprisingly high (0.293) Cry1F resistance allele frequency in a population collected in 2011 from non-Bt maize in south Florida. Field populations from non-Bt maize in 2012–2013 exhibited 18.8-fold to >85.4-fold resistance to purified Cry1F protein and those collected from unexpectedly damaged Bt maize plants at several locations in Florida and North Carolina had >85.4-fold resistance. In addition, reduced efficacy and control failure of Cry1F maize against natural populations of S. frugiperda were documented in field trials using Cry1F-based and pyramided Bt maize products in south Florida. The Cry1F-resistant S. frugiperda also showed a low level of cross-resistance to Cry1A.105 and related maize products, but not to Cry2Ab2 or Vip3A. The occurrence of Cry1F resistance in the U.S. mainland populations of S. frugiperda likely represents migration of insects from Puerto Rico, indicating the great challenges faced in achieving effective resistance management for long-distance migratory pests like S. frugiperda.

Effects of cultivation and return of Bacillus thuringiensis (Bt) maize on the diversity of the arbuscular mycorrhizal community in soils and roots of subsequently cultivated conventional maize

Cultivation of genetically modified maize (Zea mays L.) expressing the Cry1Ab protein from Bacillus thuringiensis (Bt) has increased worldwide since Bt maize was first commercialised. However, the cultivation and return of Bt maize has been shown to affect nontarget symbiotic soil-borne microbes such as arbuscular mycorrhizal fungi (AMF). In this study, we compared the diversity and composition of the AMF communities between two Bt maize plants (5422Bt1 [event Bt 11] and 5422CBCL [event MON 810]) and their conventional (non-Bt) isoline (5422) after cultivation for five seasons and return of straws by using molecular approaches, including terminal restriction fragment length polymorphism and DNA sequencing. Our data revealed that the diversities of AMF communities did not consistently differ significantly in soils and roots of subsequently planted conventional maize (SCM 5422) grown with Bt maize straw at three sampling stages (seedling, large bell, and maturity stages). DNA sequencing showed that typical AMF communities included Glomus, Paraglomus, Diversispora, Acaulospora, and Rhizophagus, of which Glomus was the most abundant. Funneliformis was detected only in bulk and rhizospheric soils and in roots of maize at the seedling stage. Rhizophagus was detected only in rhizospheric soils and only at the maturity stage. No significant effects related to the presence of Bt maize straw (5422Bt1 or 5422CBCL) were found by general linear analysis. However, plant growth stage had a greater influence on AMF diversity than Bt traits. In conclusion, cultivation of non-Bt maize on soils previously cultivated with Bt maize for five seasons had minor effects on AMF communities.

Comparative Diversity of Arthropods on Bt Maize and Non-Bt Maize in two Different Cropping Systems in South Africa

The biodiversity of an agroecosystem is not only important for its intrinsic value but also because it influences ecological functions that are vital for crop production in sustainable agricultural systems and the surrounding environment. A concern about genetically modified (GM) crops is the potential negative impact that such crops could have on diversity and abundance of nontarget organisms, and subsequently on ecosystem functions. Therefore, it is essential to assess the potential environmental risk of the release of a GM crop and to study its effect on species assemblages within that ecosystem. Assessment of the impact of Bt maize on the environment is hampered by the lack of basic checklists of species present in maize agroecosystems. The aims of the study were to compile a checklist of arthropods that occur on maize in South Africa and to compare the diversity and abundance of arthropods and functional groups on Bt maize and non-Bt maize. Collections of arthropods were carried out during two growing seasons on Bt maize and non-Bt maize plants at two localities. Three maize fields were sampled per locality during each season. Twenty plants, each of Bt maize and non-Bt maize, were randomly selected from the fields at each site. The arthropods collected during this study were classified to morphospecies level and grouped into the following functional groups: detritivores, herbivores, predators, and parasitoids. Based on feeding strategy, herbivores and predators were further divided into sucking herbivores or predators (piercing-sucking mouthparts) and chewing herbivores or predators (chewing mouthparts). A total of 8,771 arthropod individuals, comprising 288 morphospecies and presenting 20 orders, were collected. Results from this short-term study indicated that abundance and diversity of arthropods in maize and the different functional guilds were not significantly affected by Bt maize, either in terms of diversity or abundance.

Efficacy Evaluation of Two Transgenic Maize Events Expressing Fused Proteins to Cry1Ab-Susceptible and -Resistant <I>Ostrinia furnacalis</I> (Lepidoptera: Crambidae)

The Asian corn borer, Ostrinia furnacalis (GuenCe), is a major pest of maize in China. Transgenic Bt maize could provide an effective way to control this pest. However, the potential of resistance evolution has been documented in the laboratory-a CrylAb-resistant strain of Asian corn borer (Asian corn borer-AbR) could survive well on two Bt maize events, that is, MON810 and Bt11. Pyramided transgenic crops with multiple dissimilar Bt insecticidal proteins offer a superior route for pests control compared with that with one Bt single gene. In this study, two novel transgenic Bt maize events (N30 and V3), which contained a fused gene Cry1Ab/Cry2Aj and CrylAb /Vip3DA, respectively, were evaluated for their efficacy on protection against CrylAb-susceptible Asian corn borer (Asian corn borer-BtS) as well as their potential for dealing with the Asian corn borer-AbR. In laboratory bioassay, no neonate larvae of Asian corn borer-BtS fed on the whorl leaves, tassels, husks, silks, and kernels of N30 and V3 could survive at 96 h after infestation. In contrast, only 2.0-8.7% larvae died as they fed on non-Bt maize tissues. As Asian corn borer-AbR neonate larvae fed on N30 and V3 silks and kernels, 95.0% even more larvae were killed at 96 h after infestation, and all died at 168 h after infestation, while 10.0-11.67% larvae survived on non-Bt control. In the field test under artificial infestation at middle whorl leaf and silking stages of maize, there were no significant leaf, silk, and stalk damage on Bt maize plants by comparison with severe leaf feeding, stalk born, and ear damage in control, in addition, the expression profiles of a fused protein of CrylAb/Cry2Aj or CrylAb/Vip3DA in different tissues of N30 and V3 were also confirmed. In conclusion, it is clear that both Bt maize events (N30 and V3) show great potential for protecting maize from attack by Asian corn borer that has resistance to Cry1Ab protein.

Positive effect of Cry1Ab-expressing Bt maize on the development and reproduction of the predator Orius majusculus under laboratory conditions

The effects on the reproduction and nymphal development of the predator Orius majusculus Reuter (Hemiptera: Anthocoridae) caused by feeding on Bt plant materials and on herbivore prey fed on Bt plants were examined in three experiments. First, to measure the effects of Bt maize plants on O. majusculus fecundity and fertility, nymphs were provided with combinations of Bt or non-Bt pollen and leaf in addition to prey, and egg laying and egg hatching in the resulting adults were measured. Second, the effect of Bt vs. non-Bt pollen and leaf with no additional prey on nymphal development, survival, sex ratio and teneral adult weight and size were measured. Third, prey-mediated effects of Bt protein on nymphal developmental time, survival, sex ratio and teneral adult weight were evaluated using Tetranichus urticae Koch (Acarina: Tetranychidae) fed on Bt and non-Bt maize plants as prey.It has been confirmed that ingestion of Bt protein by O. majusculus via plant leaves or pollen or via the food web has no negative effects on predator survival, development, fecundity and fertility. On the contrary, in such circumstances a positive effect on fecundity and developmental time of the predator was observed. Fecundity was increased when they feed on Bt plant material, and nymphal development was shortened when nymphs were fed on Bt-containing spider mites and when they were fed on Bt plant material in the absence of lepidopteran eggs prey.

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

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 F1/F2 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.