Bt Insecticidal Proteins Research Articles

The Impact of Environmental Stress on Bt Crop Performance.

Bt crops have been grown commercially for more than two decades. They have proven remarkably effective in the control of target insect pests. However, Bt crops can become less effective under various forms of environmental stress. Most studies in this area have considered the effect of environmental stress on Bt insecticidal protein levels or target pest mortality, but not both, resulting in a lack of mechanistic analysis. In this review, we critically examine previous research addressing the impact of environmental stress on the effectiveness of Bt crops. We find that the body of research data is not sufficiently robust to allow the reliable prediction of the performance of Bt crops under extreme climatic conditions.

Assessment of Pigeonpea (Cajanus cajan L.) transgenics expressing Bt ICPs, Cry2Aa and Cry1AcF under nethouse containment implicated an effective control against herbivory by Helicoverpa armigera (Hübner).

Pigeonpea is a source of quality proteins and the main constituent of a well-balanced diet for majority of Indian population. One of the major constraints in the production of pigeonpea is a polyphagous insect pest, Helicoverpa armigera. Non-availability of resistant sources in the germplasm and limitations in conventional breeding have been key factors for continued yield losses. Additionally, hazards of chemical fertilizers on the environment have prompted the scientific community to develop alternative strategies. Bacillus thuringiensis (Bt) insecticidal proteins (ICPs) have emerged as the most reliable source for the control of insect pests through transgenics. Transgenic pigeonpea plants harboring validated Bt ICPs, Cry2Aa and Cry1AcF were developed by a non-tissue culture based in planta transformation strategy and assessed for integration of Transfer-DNA (T-DNA) and efficacy against pod borer under in vitro conditions. For the first time this study demonstrates the successful evaluation of 19 transgenic pigeonpea events (11 with cry2Aa and 8 with cry1AcF) under soil and pot conditions in a nethouse containment. The stability in the performance was assessed stringently by deliberate H. armigera larval challenging. The trial identified ten promising events of both the genes that portrayed reduced damage to the herbivore. We present the first ever successful evaluation of pigeonpea transgenics with the ability to mitigate pod borer under nethouse conditions. The transgenics depicted molecular evidence for the stability of T-DNA integration, consistency in the expression of Cry proteins and resistance against H. armigera. These events can form a pool of useful transgenics to manage the devastating pod borer. © 2019 Society of Chemical Industry.

Impact of Various Bt Cotton Traits and the Application of an Insecticide on the Within Plant Distribution of Helicoverpa zea (Lepidoptera: Noctuidae) Larvae and Injured Floral Structures

Previous studies have indicated that the expression of insecticidal proteins from the bacterium Bacillus thuringiensis (Bt) in cotton can have a significant influence on the behavior of bollworm larvae (Helicoverpa zea, Lepidoptera:Noctuidae). This suggests that the particular Bt protein produced by a cotton variety may need to be considered when determining the most ideal scouting methods to utilize for bollworm. NonBt, WideStrike (producing Cry1Ac + Cry1F Bt insecticidal proteins), and Bollgard II (Cry1Ac + Cry2Ab) cotton varieties were planted and either treated with an insecticide or left untreated. The presence of H. zea larvae and their feeding injury were recorded according to their location in the canopy and type of floral structure where they were found. Results from comparison of larval and injury distributions indicated no significant differences between the different cotton varieties tested, and that insecticide treatment had minimal impact on this distribution. Larval size was generally associated with location in the canopy, suggesting that larvae tend to move towards the middle of the canopy as they age. The effect of different Bt cotton technologies appears to associate with how quickly larvae move to preferred feeding sites rather than their preference for particular feeding sites. These results suggest that scouting methods could be standardized independently of the presence of a Bt cotton trait or previous insecticide application. Focusing scouting efforts on the middle portion of the canopy (i.e., nodes 6-9) should increase the detection of small larvae and ‘fresh’ injury and be less influenced by previous insecticide applications.

Performance of Field-Collected Spodoptera frugiperda (Lepidoptera: Noctuidae) Strains Exposed to Different Transgenic and Refuge Maize Hybrids in Argentina.

Spodoptera frugiperda (J. E. Smith) is one of the major pests of maize in Argentina. The main tool for its control is the use of genetically modified maize hybrids expressing Bacillus thuringiensis (Bt) insecticidal proteins. Maize growers in Argentina initially controlled this pest with Bt maize that expressed a single Bt protein (Cry1Ab or Cry1F). Currently it is necessary to plant maize cultivars that produce two Bt proteins to achieve the satisfactory control. Recently, Cry1F field-evolved resistant populations of this species were confirmed in Argentina. The objective of this study was to evaluate the performance of S. frugiperda field-collected strains on different Bt and non-Bt maize hybrids. Strains were collected from non-Bt maize (T1), Agrisure TDMax (T2), Agrisure Viptera (T3), Agrisure Viptera 3110 (T4), Genuity VT Triple Pro (T5), and Power Core (T6). Three experiments were performed to 1) determine the survivorship and reproduction of field-collected larvae (F0) from Bt maize hybrids, 2) evaluate Cry1F resistance using an F1 screen, and 3) assess the performance of F1 strains on different maize hybrids. In the F0, the survivorship from larva to adult ranged from 0 to 63%. We obtained adults from only the T1, T2, T5, and T6 strains with no significant differences in the reproductive parameters. Continuously rearing F1 larvae on their collected hosts affected larval duration, which was significantly shorter for a known-laboratory Bt-susceptible strain than the field-collected strains. Our results support the existence of Cry1F-resistance alleles in S. frugiperda field populations in Argentina.

Recovery characteristics of Bt insecticidal protein and relative physiological mechanisms after high temperature stress termination in square of Bt cotton

The experiments were conducted at Key Laboratory of Genetics and Physiology of Yangzhou University. The conventional cultivar Sikang-1 and hybrid cultivar Sikang-3 were used as the experimental materials. The potted cotton at squaring stage was moved to artificial climatic chamber with 38°C for 72 h , and 38°C for 96 h respectively in 2017 and 2018 cotton growth season, and moved to natural conditions, then the squares were collected to measure the Bt protein concentration and nitrogen metabolic physiology at 0, 12, 24, 48, 72, 96 hours respectively after the high temperature termination. The square Bt insecticidal protein contents were able to recover to the corresponding control level, the time for the recovery was in need of longer as the stressed high temperature period increased. In 72 h high temperature treatment, the square Bt insecticidal protein content could recover to the corresponding control level at 72 h for Sikang-1 and at 48 h for Sikang-3. In 96 h high temperature treatment, the square Bt insecticidal protein content could recover to the corresponding control level at 96 h for Sikang-1 and at 72 h for Sikang-3. There were significantly positive correlations of Bt toxin content with soluble protein content, glutamate pyruvate transaminase activity and glutamate oxaloacetate transaminase activity (the correlation coefficients were 0.964**, 0.981**, and 0.971**, respectively), and significantly negative correlations of Bt toxin content with free amino acid contents, and activitied of protease and peptidase (the correlation coefficients were -0.894**, -0.912**, and -0.834**, respectively). Therefore, the recovery degree of Bt insecticidal protein content of cotton square can be predicted according to the duration of high temperature stress, reasonably controlling Helicoverpa armigera and related pests in production.

Experimental and theoretical landscape influences on Spodoptera frugiperda movement and resistance evolution in contaminated refuge areas of Bt cotton

Transgenic cotton plants producing Bacillus thuringiensis (Bt) (Berliner) insecticidal proteins have contributed to the management of key lepidopteran pests. Larval high dispersal rates between non-Bt and Bt plants in landscapes with seed contamination can speed the evolution of insect resistance to Bt plants. We evaluated the effect of Bt and non-Bt cotton plants on the larval dispersal pattern and survival of susceptible, heterozygous and Cry1F-resistant Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) genotypes in pure and contaminated artificial micro-landscape. A computer model was used to analyse the consequences of S. frugiperda larval dispersal behaviour on resistance evolution in refuge areas with different contamination levels and migration of different adult genotype combinations. The biological data from artificial micro-landscape experiments were used in simulations of macro-landscape scales. The Cry1F-resistant genotype avoided non-Bt cotton. The heterozygote had a similar larval dispersal behaviour as the susceptible genotype when non-Bt cotton was the central plant. Our simulations provide evidence that in refuge areas contaminated with Bt cotton plants, the evolution of resistance may be > 75-fold faster in relation to a contamination-free refuge. In conclusion, S. frugiperda resistance management practices on regional scales with contamination-free refuges are important to prevent loss in different crops.

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.

First record of Peridroma saucia Hübner (Lepidoptera: Noctuidae) in transgenic soybeans

The widespread cultivation of transgenic soybeans has caused significant changes in the spectrum of Lepidoptera larvae, both in the number of species as well as on their densities in the field. Transgenic crops producing Bacillus thuringiensis (Bt) insecticidal proteins have successfully reduced the incidence of the most common caterpillars infesting soybeans, namely Anticarsia gemmatalis (Lepidoptera: Erebidae) and Chrysodeixis includens (Lepidoptera: Noctuidae). However, lepidopteran species not previously recorded on the crop have been recently found, and are of concern due to the possibility of adaptation to the genetically modified cultivars. The occurrence of Peridroma saucia Hübner (Lepidoptera: Noctuidae) is described for the first time in Brazil feeding on genetically modified soybean cultivars.

Molecular and structural characterization of a novel Cry1D toxin from Bacillus thuringiensis with high toxicity to Spodoptera littoralis (Lepidoptera: Noctuidae)

The investigation of new Bacillus thuringiensis (Bt) insecticidal proteins (Cry) with specific toxicity is one of the alternative measures used for Lepidopteran pest control. In the present study, a new Cry toxin was identified from a promising Bt strain BLB250 which was previously selected for its high toxicity against Spodoptera littoralis. The corresponding gene, designated cry1D-250, was cloned. It showed an ORF of 3498bp, encoding a protein of 1165 amino acid residues with a putative molecular mass of 132kDa which was confirmed by SDS-PAGE and Western blot analyses. The corresponding toxin named Cry1D-250 showed a higher insecticidal activity towards S. littoralis than Cry1D-133 (LC50 of 224.4ngcm−2) with an LC50 of only 166ngcm−2. Besides to the 65kDa active toxin, proteolysis activation of Cry1D-133 protein with S. littoralis midgut juice generated an extra form of 56kDa, which was the result of a second cleavage. Via activation study and 3D structure analysis, novel substitutions found in the Cry1D-250 protein compared to Cry1D-133 toxin were shown to be involved in the protein stability and toxicity. Therefore, the Cry1D-250 toxin can be considered to be an effective alternative for the control of S. littoralis.

Susceptible and mCry3A resistant corn rootworm larvae killed by a non-hemolytic Bacillus thuringiensis Cyt1Aa mutant

The western corn rootworm (WCR) Diabrotica virgifera virgifera causes substantial damage in corn. Genetically modified (GM) plants expressing some Bacillus thuringiensis (Bt) insecticidal Cry proteins efficiently controlled this pest. However, changes in WCR susceptibility to these Bt traits have evolved and identification of insecticidal proteins with different modes of action against WCR is necessary. We show here for the first time that Cyt1Aa from Bt exhibits toxicity against WCR besides to the dipteran Aedes aegypti larvae. Cyt1Aa is a pore-forming toxin that shows no cross-resistance with mosquitocidal Cry toxins. We characterized different mutations in helix α-A from Cyt1Aa. Two mutants (A61C and A59C) exhibited reduced or absent hemolytic activity but retained toxicity to A. aegypti larvae, suggesting that insecticidal and hemolytic activities of Cyt1Aa are independent activities. These mutants were still able to form oligomers in synthetic lipid vesicles and to synergize Cry11Aa toxicity. Remarkably, mutant A61C showed a five-fold increase insecticidal activity against mosquito and almost 11-fold higher activity against WCR. Cyt1Aa A61C mutant was as potent in killing WCR that were selected for resistance to mCry3A as it was against unselected WCR indicating that this toxin could be a useful resistance management option in the control of WCR.

Effects of straw leachates from Cry1C-expressing transgenic rice on the development and reproduction of Daphnia magna

The transgenic rice line T1C-19 provides high resistance to lepidopteran pests because of the synthesis of the Bacillus thuringiensis (Bt) insecticidal protein Cry1C. It thus shows good prospect for commercial planting in China. Species of Cladocera, an order of aquatic arthropods commonly found in aquatic ecosystems such as rice paddies, might be exposed to the insecticidal protein released from Bt-transgenic rice-straw residues. For the study reported herein, we used Daphnia magna (water flea) as a representative of Cladocera to evaluate whether aquatic arthropods are adversely affected when exposed to Bt rice-straw leachates. We exposed D. magna to M4 medium containing various volume percentages of medium that had been incubated with T1C-19 rice straw or rice straw from its non-transformed near-isoline Minghui 63 (MH63) for 21 days. Compared with pure M4 medium (control), the fitness and developmental and reproduction parameters of D. magna decreased significantly when exposed to rice-straw leachates; conversely, no significant differences between the T1C-19 and MH63 rice-straw leachate treatments were observed, indicating that the Bt rice straw leachate did not adversely affect this non-target species.

ABC transporter mis-splicing associated with resistance to Bt toxin Cry2Ab in laboratory- and field-selected pink bollworm

Evolution of pest resistance threatens the benefits of genetically engineered crops that produce Bacillus thuringiensis (Bt) insecticidal proteins. Strategies intended to delay pest resistance are most effective when implemented proactively. Accordingly, researchers have selected for and analyzed resistance to Bt toxins in many laboratory strains of pests before resistance evolves in the field, but the utility of this approach depends on the largely untested assumption that laboratory- and field-selected resistance to Bt toxins are similar. Here we compared the genetic basis of resistance to Bt toxin Cry2Ab, which is widely deployed in transgenic crops, between laboratory- and field-selected populations of the pink bollworm (Pectinophora gossypiella), a global pest of cotton. We discovered that resistance to Cry2Ab is associated with mutations disrupting the same ATP-binding cassette transporter gene (PgABCA2) in a laboratory-selected strain from Arizona, USA, and in field-selected populations from India. The most common mutation, loss of exon 6 caused by alternative splicing, occurred in resistant larvae from both locations. Together with previous data, the results imply that mutations in the same gene confer Bt resistance in laboratory- and field-selected strains and suggest that focusing on ABCA2 genes may help to accelerate progress in monitoring and managing resistance to Cry2Ab.

Cry78Aa, a novel Bacillus thuringiensis insecticidal protein with activity against Laodelphax striatellus and Nilaparvata lugens.

Transgenic plants expressing insecticidal proteins originating from Bacillus thuringiensis (Bt) have successfully been used to control lepidopteran and coleopteran pests with chewing mouthparts. However, only a handful of Bt proteins have been identified that have bioactivity against sap sucking pests (Hemiptera), including aphids, whiteflies, plant bugs and planthoppers. A novel Bt insecticidal protein with significant toxicity against a hemipteran insect pest is described here. The gene encoding the 359 amino acid, 40.7 kDa protein was cloned from strain C9F1. After expression and purification of the toxin, its median lethal concentration (LC50) values against Laodelphax striatellus and Nilaparvata lugens were determined as 6.89 μg/mL and 15.78 μg/mL respectively. Analysis of the toxin sequence revealed the presence of both Toxin_10 and Ricin_B_Lectin domains.

Safety Assessment of Bacillus thuringiensis Insecticidal Proteins Cry1C and Cry2A with a Zebrafish Embryotoxicity Test.

As a result of the large-scale planting of transgenic Bacillus thuringiensis (Bt) crops, fish would be exposed to freely soluble Bt insecticidal protein(s) that are released from Bt crop tissues into adjacent bodies of water or by way of direct feeding on deposited plant material. To assess the safety of two Bt proteins Cry1C and Cry2A to fish, we used zebrafish as a representative species and exposed their embryos to 0.1, 1, and 10 mg/L of the two Cry proteins until 132 h post-fertilization and then several developmental, biochemical, and molecular parameters were evaluated. Chlorpyrifos (CPF), a known toxicant to aquatic organisms, was used as a positive control. Although CPF exposure resulted in significant developmental, biochemical, and molecular changes in the zebrafish embryos, there were almost no significant differences after Cry1C or Cry2A exposure. Thus, we conclude that zebrafish embryos are not sensitive to Cry1C and Cry2A insecticidal proteins at test concentrations.

Establishment of a dietary exposure assay for evaluating the toxicity of insecticidal compounds to Apolygus lucorum (Hemiptera: Miridae).

With the commercialization of transgenic cotton that expresses Bt (Bacillus thuringiensis) insecticidal proteins, mirid bugs have become key pests in cotton and maize fields in China. Genetically engineered (GE) crops for controlling mirids are unavailable owing to a lack of suitable insecticidal genes. In this study, we developed and validated a dietary exposure assay for screening insecticidal compounds and for assessing the potential effects of insecticidal proteins produced by GE plants on Apolygus lucorum, one of the main mirid pests of Bt cotton and Bt maize. Diets containing potassium arsenate (PA) or the cysteine protease inhibitor E-64 were used as positive controls for validating the efficacy of the dietary exposure assay. The results showed that with increasing concentrations of PA or E-64, A.lucorum larval development time was prolonged and adult weight and fecundity were decreased, suggesting that the dietary exposure assay was useful for detecting the toxicity of insecticidal compounds to A.lucorum. This assay was then used to assess the toxicity of Cry1Ab, Cry1Ac, Cry1F, Cry2Aa, and Cry2Ab proteins, which have been transformed into several crops, against A.lucorum. The results showed that A.lucorum did not show a negative effect by feeding on an artificial diet containing any of the purified Cry proteins. No significant changes in the activities of digestive, detoxifying, or antioxidant enzymes were detected in A.lucorum that fed on a diet containing Cry proteins, but A.lucorum fitness was reduced when the insect fed on a diet containing E-64 or PA. These results demonstrate that A.lucorum is not sensitive to the tested Cry proteins and that the dietary exposure assay is useful for evaluating the toxicity of insecticidal compounds to this species.

Field-evolved resistance to Bt maize in sugarcane borer (Diatraea saccharalis) in Argentina.

BACKGROUNDMaize technologies expressing Bacillus thuringiensis (Bt) insecticidal proteins are widely used in Argentina to control sugarcane borer (Diatraea saccharalis Fabricius). Unexpected D. saccharalis damage was observed to Bt maize events TC1507 (expressing Cry1F) and MON 89034 × MON 88017 (expressing Cry1A.105 and Cry2Ab2) in an isolated area of San Luis Province. Diatraea saccharalis larvae were sampled from MON 89034 × MON 88017 fields in the area to generate a resistant strain (RR), which was subsequently characterized in plant and diet bioassays.RESULTSSurvivorship of the RR strain was high on TC1507 leaf tissue, intermediate on MON 89034 × MON 88017, and low on MON 810 (expressing Cry1Ab). The RR strain had high resistance to Cry1A.105 (186.74‐fold) and no resistance to Cry2Ab2 in diet bioassays. These results indicate resistance to Cry1F and Cry1A.105 (and likely cross‐resistance between them) but not to Cry1Ab or Cry2Ab2. Resistance to MON 89034 × MON 88017 was functionally recessive. Reviews of grower records suggest that resistance initially evolved to Cry1F, conferring cross‐resistance to Cry1A.105, with low refuge compliance as the primary cause. A mitigation plan was implemented in San Luis that included technology rotation, field monitoring, and grower education on best management practices (BMPs) including refuges.CONCLUSIONIn the affected area, the resistance to Cry1F and Cry1A.105 is being managed effectively through use of MON 89034 × MON 88017 and MON 810 in combination with BMPs, and no spread of resistance to other regions has been observed. © 2017 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Safety of Bacillus thuringiensis Cry1C protein for Daphnia magna based on different functional traits

Cry1C is a Bacillus thuringiensis (Bt) insecticidal protein and it can be produced by transgenic rice lines developed in China. Cladocera species are common aquatic arthropods that may be exposed to insecticidal proteins produced in Bt-transgenic plants through ingestion of pollen or crop residues in water. As the cladoceran Daphnia magna plays an important role in the aquatic food chain, it is important to assess the possible effects of Bt crops to this species. To evaluate the safety of the Cry1C protein for D. magna, individuals were exposed to different concentrations of purified Cry1C protein in M4 medium for 21 days. Potassium dichromate (K2Cr2O7), a known toxicant to D. magna, was added to M4 medium as a positive control treatment, and pure M4 medium was used as a negative control. Our results show that developmental, reproductive, and biochemical parameters of D. magna were not significantly different between Cry1C and negative control treatments but were significantly inhibited by the positive control. We thus conclude that D. magna is insensitive to Cry1C.

A selective insecticidal protein from Pseudomonas mosselii for corn rootworm control.

SummaryThe coleopteran insect western corn rootworm (WCR, Diabrotica virgifera virgifera) is an economically important pest in North America and Europe. Transgenic corn plants producing Bacillus thuringiensis (Bt) insecticidal proteins have been useful against this devastating pest, but evolution of resistance has reduced their efficacy. Here, we report the discovery of a novel insecticidal protein, PIP‐47Aa, from an isolate of Pseudomonas mosselii. PIP‐47Aa sequence shows no shared motifs, domains or signatures with other known proteins. Recombinant PIP‐47Aa kills WCR, two other corn rootworm pests (Diabrotica barberi and Diabrotica undecimpunctata howardi) and two other beetle species (Diabrotica speciosa and Phyllotreta cruciferae), but it was not toxic to the spotted lady beetle (Coleomegilla maculata) or seven species of Lepidoptera and Hemiptera. Transgenic corn plants expressing PIP‐47Aa show significant protection from root damage by WCR. PIP‐47Aa kills a WCR strain resistant to mCry3A and does not share rootworm midgut binding sites with mCry3A or AfIP‐1A/1B from Alcaligenes that acts like Cry34Ab1/Cry35Ab1. Our results indicate that PIP‐47Aa is a novel insecticidal protein for controlling the corn rootworm pests.

Bt Proteins Have No Detrimental Effects on Larvae of the Green Lacewing, Chrysopa pallens (Rambur) (Neuroptera: Chrysopidae)

Biosafety of a genetically modified crop is required to be assessed prior to its commercialization. For this, a suitable artificial diet was developed and used to establish a dietary exposure test for assessing the toxicity of midgut-active Bt insecticidal proteins on Chrysopa pallens (Rambur). Subsequently, this dietary exposure test was used to evaluate the toxicity of the proteins Cry1Ab, Cry1Ac, Cry1Ah, Cry1Ca, Cry1F, Cry2Aa, Cry2Ab, and Vip3Aa on C. pallens larvae. Temporal stability, bioactivity, and the intake of the insecticidal proteins were confirmed by enzyme-linked immunosorbent assay and a sensitive-insect bioassay. The life history characteristics, such as survival, pupation, adult emergence, 7-day larval weight, larval developmental time, and emerged male and female fresh weights remained unaffected, when C. pallens were fed the pure artificial diet (negative control) and the artificial diets containing 200μg/g of each purified protein: Cry1Ab, Cry1Ac, Cry1Ah, Cry1Ca, Cry1F, Cry2Aa, Cry2Ab, or Vip3Aa. On the contrary, all of the life history characteristics of C. pallens larvae were adversely affected when fed artificial diet containing boric acid (positive control). The results demonstrate that diets containing the tested concentrations of Cry1Ab, Cry1Ac, Cry1Ah, Cry1Ca, Cry1F, Cry2Aa, Cry2Ab, and Vip3Aa have null effects on C. pallens larvae. The outcome indicates that genetically modified crops expressing the tested Bt proteins are safe for the lacewing, C. pallens.

Susceptibility ofSpodoptera frugiperda(Lepidoptera: Noctuidae) Field Populations to the Cry1FBacillus thuringiensisInsecticidal Protein

The fall armyworm, Spodoptera frugiperda (Smith & Abbot) (Lepidoptera: Noctuidae), is a polyphagous insect pest affecting multiple crops. Fall armyworm in corn is managed with insecticides and corn hybrids expressing insecticidal proteins derived from Bacillus thuringiensis Berliner (Bt). The early detection of insect resistance is important for making appropriate management decisions and for implementing integrated pest management and insect resistance management recommendations. The objective of this study was to estimate susceptibility of fall armyworm populations to the Cry1F Bt insecticidal protein, emphasizing collections from locations where fall armyworm overwinters in the U.S. Fall armyworm neonates were exposed to artificial diet treated with increasing Cry1F concentrations, and mortality and growth inhibition were evaluated after 7 d. Differences in Cry1F susceptibility between the most susceptible and the most tolerant field populations were 2- and 6-fold for 2012 and 2013, respectively. These results are consistent with other susceptibility studies of Bt toxicity in other species although reduced susceptibility in one population collected from Florida may suggest resistance development.