Cry1F Protein Research Articles

Toxicity Analysis of Local Isolates of Bacillus thuringiensis and CRY1F Protein against Selected Lepidoptera Pests

Toxicity Analysis of Local Isolates of Bacillus thuringiensis and CRY1F Protein against Selected Lepidoptera Pests

Side Effects of Single-Transgene or Pyramided Genetically Modified Maize on the Generalist Endoparasitoid Palmistichus elaeisis (Hymenoptera: Eulophidae)

The fall armyworm, Spodoptera frugiperda J.E. Smith (Lepidoptera: Noctuidae), is an agricultural pest native to tropical and subtropical regions of the Americas, with considerable potential for global invasion and seasonal migration. Although genetically modified (GM) plants have shown positive impacts on the economy and the environment as they synthesize proteins that act as natural insecticides and are primarily intended to protect the crops from damage by specific pests, potential effects of Bt toxins on non-target organisms can occur. This experiment aimed to evaluate the potential impacts on the parasitoid Palmistichus elaeisis (Hymenoptera: Eulophidae), using the pupae of Spodoptera frugiperda (Lepidoptera: Noctuidae) as hosts, which were fed with transgenic maize genotypes such as Herculex®, expressing Cry1F protein, and PowerCore®, expressing Cry1F, Cry1A.105, and Cry2Ab2 proteins, or their non-transgenic isohybrid. The experiment was conducted in a completely randomized design with ten replicates. Spodoptera frugiperda larvae were fed ad libitum with transgenic and non-transgenic maize until the pupal stage and then individually exposed to six P. elaeisis females for 78 h. The total number of adults, emerged males, tibia size, cephalic capsule size, and parasitoid body size were not influenced by host feeding. However, the number of emerged females from the Herculex® and PowerCore® treatments was lower than that for the isohybrid treatment. The sex ratio and longevity of parasitoids emerging from PowerCore® were 1.05 and 1.26 times lower, respectively, than that of those from the isohybrid. Furthermore, the number of dead P. elaeisis within the host pupa was 10.56 times higher in the PowerCore® genotype. Combining Bt proteins within pyramided genotypes should cause minimal impacts and promote the conservation and integration of beneficial insects. The results of this study provide helpful information for developing effective and compatible integrated pest management (IPM) strategies.

Shared genetic basis of resistance to Cry1F protein in three independent Brazilian strains of fall armyworm

Field strains of the fall armyworm (FAW), Spodoptera frugiperda (Lepidoptera: Noctuidae), a global pest of corn, have evolved practical resistance to transgenic corn-producing the Bt protein Cry1F in Brazil, Argentina, and the United States. Considering the rapid invasion of FAW and the worldwide adoption of Bt crops for its control, it is important to understand the genetic basis of resistance to Cry1F in resistant strains of FAW from different regions of the world. This information is essential for developing new transgenic Bt crops and designing appropriate strategies for resistance management for FAW in Bt crops. Thus, the objective of this work was to determine whether Brazilian strains of FAW from different geographic origins share the same locus of resistance to Cry1F. We conducted interstrain complementation tests for allelism among three resistant strains. The survival of F1 progenies from crosses among FAW-resistant strains in purified Cry1F protein or Cry1F-corn was similar to that observed for the parental strains. Our results suggest that mutations conferring resistance to Cry1F Bt protein in three strains of FAW are present in a single locus. Results from this study provide information for developing effective strategies for Bt resistance management, which is the main limitation for sustainable use of Bt technology against this pest.

Performance of cotton expressing Cry1Ac, Cry1F and Vip3Aa19 insecticidal proteins against Helicoverpa armigera, H. zea and their hybrid progeny, and evidence of reduced susceptibility of a field population of H. zea to Cry1 and Vip3Aa in Brazil.

The genetically modified cotton DAS-21023-5 × DAS-24236-5 × SYN-IR102-7 expressing Cry1Ac, Cry1F and Vip3Aa19 from Bacillus thuringiensis Berliner (Bt) has been cultivated in Brazil since the 2020/2021 season. Here, we assessed the performance of DAS-21023-5 × DAS-24236-5 × SYN-IR102-7 cotton expressing Cry1Ac, Cry1F and Vip3Aa19 against Helicoverpa armigera (Hübner), Helicoverpa zea (Boddie), and their hybrid progeny. We also carried out evaluations with DAS-21023-5 × DAS-24236-5 cotton containing Cry1Ac and Cry1F. In leaf-disk bioassays, DAS-21023-5 × DAS-24236-5 × SYN-IR102-7 was effective in controlling neonates from laboratory colonies of H. armigera, H. zea and the hybrid progeny (71.9%-100% mortality). On floral bud bioassays using L2 larvae, H. zea presented complete mortality, whereas H. armigera and the hybrid progeny showed <55% mortality. On DAS-21023-5 × DAS-24236-5 cotton, the mortality of H. armigera on leaf-disk and floral buds ranged from 60% to 73%, whereas mortality of hybrids was <46%. This Bt cotton caused complete mortality of H. zea larvae from a laboratory colony in the early growth stages, but mortalities were <55% on advanced growth stages and on floral buds. In field studies conducted from 2014 to 2019, DAS-21023-5 × DAS-24236-5 × SYN-IR102-7 cotton was also effective at protecting plants against H. armigera. In contrast, a population of H. zea collected in western Bahia in 2021/2022 on Bt cotton expressing Cry1 and Vip3Aa proteins, showed 63% mortality after 30 d, with insects developing into fifth and sixth instars, on DAS-21023-5 × DAS-24236-5 × SYN-IR102-7 cotton. We conclude that H. armigera, H. zea, and their hybrid progeny can be managed with DAS-21023-5 × DAS-24236-5 × SYN-IR102-7 cotton; however we found the first evidence in Brazil of a significant reduction in the susceptibility to DAS-21023-5 × DAS-24236-5 × SYN-IR102-7 cotton of a population of H. zea collected from Bt cotton in Bahia in 2021/2022.

Rotation of Multiple Single-Gene Transgenic Crops Did Not Slow the Evolution of Resistance to Cry1F or Cry1Ie in Ostrinia furnacalis.

A common strategy for delaying the evolution of resistance to transgenic crops that produce insecticidal proteins from Bacillus thuringiensis is to ensure that insect pests are exposed to multiple toxins with different mechanisms of action (MoAs). This can take the form of planting crops in a rotation pattern when different crops expressing single toxins are available on the market. The efficacy of a rotation strategy is reliant on mathematical models based on biological assumptions. Here, we designed laboratory evolution experiments to test whether Bt-based insecticidal proteins with different MoAs used in rotation could delay resistance from developing in Asian corn borer (ACB), Ostrinia furnacalis. We investigated the proteins Cry1Ab, Cry1F, and Cry1Ie, which are widely utilized for commercial insect control. We found that rotation of multiple toxins did not slow the evolution of resistance to Cry1F or Cry1Ie. Furthermore, the evolution of ACB to the Cry1Ab toxin develops faster when Cry1F or Cry1Ie is present, as compared to Cry1Ab exposure only. Our results suggest that toxins used in a rotation fashion do not work as an effective strategy in delaying ACB resistance evolution to Cry toxins over one-toxin exposure. Our result highlights the need to better understand the biological factors leading to insecticidal protein resistance and to develop IRM strategies against target insects.

Equivalence of grain and forage composition in corn hybrid (Zea mays L.) from genetically modified off-patent (event TC1507) and non-genetically modified conventional corn

Off-patent corn (event TC1507) contains genes coding for CRY1F and PAT proteins, which confer resistance to lepidopteran insects and tolerance to the herbicide glufosinate-ammonium. We employed the substantial equivalence approach to investigate the compositional safety of the corn off-patent event (TC1507). The levels of the nutritional contents of proximate analytes in grain and forage tissues of off-patent genotypes of transgenic maize plants and conventional corn genotypes were compared. The levels of the analytes evaluated in the transgenic plants were found to be within the ranges published in the literature for non-transgenic corn and were statistically indistinguishable from the conventional corn from which they are derived (elite corn lines), indicating substantial equivalence between the off-patent (event TC1507) and its conventional counterpart. These results constitute key evidence of the safety evaluation of the world’s first transgenic corn developed from technologies that are in the public domain.

Assessment of the potential toxicity of insecticidal compounds to Peristenus spretus, a parasitoid of mirid bugs

With the increased cultivation of Bt crops in China, Apolygus lucorum and other mirid bugs have emerged as important agricultural pests because they are insensitive to the Bt proteins. In addition, the reduction of pesticide applications after planting Bt crops also increases the severity of mirid bug outbreaks. Peristenus spretus is a parasitoid of mirid nymphs, but its sensitivity to Bt proteins is not known. In the current study, we developed a dietary exposure assay to assess the effects of Bt proteins (Cry1Ab, Cry1Ac, Cry1F, Cry2Aa, and Cry2Ab) on P. spretus adults using a diet consisting of a 10% honey solution with or without Bt proteins at 400 µg g–1 diet. The results showed that the survival and reproduction of P. spretus adults were reduced by the cysteine protease inhibitor E-64 (a positive control) but were not affected by any of the five Bt proteins. The activities of digestive, detoxifying, and antioxidant enzymes in P. spretus were also unaffected by diets containing the Cry proteins, but they were significantly affected by the diet containing E-64. We then developed a tri-trophic bioassay to determine the effects of the five Bt proteins on P. spretus larvae and pupae. In this assay, A. lucorum nymphs fed an artificial diet containing Cry proteins were used as the hosts for P. spretus. The results of the tri-trophic assay indicated that neither the pupation rate nor the eclosion rate of the P. spretus parasitoids were significantly affected by the presence of high concentrations of Bt proteins in the parasitized A. lucorum nymphs. The overall results indicate that these two assays can be used to evaluate the toxicity of insecticidal compounds to P. spretus and that the tested Cry proteins are not toxic to P. spretus.

Spodoptera frugiperda (Noctuidae) fed on transgenic maize can transfer Bt proteins to Podisus nigrispinus (Pentatomidae)

An important concern with the use of genetically modified (GM) plants expressing Bacillus thuringiensis (Bt) insecticidal toxins is the deleterious effect on non–target organisms. The predatory stink bug Podisus nigrispinus (Dallas) (Hemiptera: Pentatomidae) is used in biological control programs and may be exposed to Bt toxins. This study evaluated the indirect effects of different Cry proteins on P. nigrispinus with the prey Spodoptera frugiperda (J.E. Smith, 1797) (Lepidoptera: Noctuidae), fed on simple or pyramided Bt maize genotypes. The experiment was carried out in a completely randomized design with three treatments: i) Isohybrid (not Bt), ii) Herculex® (transgenic maize encoding Cry1F protein) and iii) PowerCore® (pyramidal transgenic maize encoding the Cry1F, Cry1A.105, and Cry2Ab2 proteins), which were used to feed S. frugiperda for 48 h. The caterpillars were used as prey by P. nigrispinus females. We evaluated the presence of Cry proteins, consumed prey biomass (predation), oviposition period, number of postures, number of eggs, number of eggs per posture, number of nymphs, egg viability, embryonic period, female longevity and development, and survival rates of immature. The results show that different Cry proteins move through the food chain of P. nigrispinus and provide evidence that the ingestion of three different proteins does not lead to unexpected synergistic effects. However, Cry toxins promoted histopathological changes in midgut cells of P. nigrispinus.

Evaluation of Reference Genes and Expression Level of Genes Potentially Involved in the Mode of Action of Cry1Ac and Cry1F in a Susceptible Reference Strain of Chrysodeixis includens.

Simple SummarySoybean looper (a moth species) is a major pest of soybean plants in the American continent and its larvae need to be kept under economic damage thresholds to guarantee sustainable yields. Soybean looper control relies mostly on the use of insecticides and genetically modified crops expressing Bacillus thuringiensis (Bt) insecticidal proteins. Due to the high selection pressure exerted by these control measures, resistance has developed to different insecticides and Bt proteins. Here, we tested the basal sensitivity of a soybean looper laboratory reference strain against two insecticidal proteins and determined the level of expression of potential receptors of these proteins across all (six) larval stages. Furthermore, we identified stable reference genes across all larval stages to normalize gene expression data obtained by quantitative polymerase chain reaction (qPCR). The results presented in this communication are useful to support future studies on insecticide and insecticidal protein resistance in soybean looper.Soybean looper (SBL), Chrysodeixis includens (Walker), is one of the major lepidopteran pests of soybean in the American continent. SBL control relies mostly on the use of insecticides and genetically modified crops expressing Bacillus thuringiensis (Bt) insecticidal Cry proteins. Due to the high selection pressure exerted by these control measures, resistance has developed to different insecticides and Bt proteins. Nevertheless, studies on the mechanistic background are still scarce. Here, the susceptibility of the laboratory SBL-Benzon strain to the Bt proteins Cry1Ac and Cry1F was determined in diet overlay assays and revealed a greater activity of Cry1Ac than Cry1F, thus confirming results obtained for other sensitive SBL strains. A reference gene study across larval stages with four candidate genes revealed that RPL10 and EF1 were the most stable genes for normalization of gene expression data obtained by RT-qPCR. Finally, the basal expression levels of eight potential Bt protein receptor genes in six larval instars were analyzed, including ATP-binding cassette (ABC) transporters, alkaline phosphatase, aminopeptidases, and cadherin. The results presented here provide fundamental knowledge to support future SBL resistance studies.

Genetic basis and cross-resistance of Vip3Aa resistance in Spodoptera frugiperda (lepidoptera: Noctuidae) derived from Texas, USA

The fall armyworm, Spodoptera frugiperda (J.E. Smith), is an economically important crop pest and a major target of transgenic crops expressing Cry and/or Vip3Aa proteins. Widespread adoption of Bt crops has placed a strong selection pressure on the insect populations. Till now, seven lepidopteran and two coleopteran insect species have developed the field-evolved resistance to a Bt crop with practical consequences. Understanding the genetic basis of insect resistance to a Bt protein is essential for designing appropriate resistance management strategies. In this study, we firstly characterized the genetic basis of Vip3Aa resistance in S. frugiperda derived from Texas. Then, we compared the genetic basis of Vip3Aa resistance of this Texas population with a known Vip3Aa resistant population of S. frugiperda collected from Louisiana to determine whether the genetic basis of the resistance in these two resistant populations was the same. Finally, we evaluated the responses of the Vip3Aa-resistant population of S. frugiperda collected from Texas against Cry1F and Cry2Ab2 purified proteins. The results showed that the resistance to Vip3Aa protein in S. frugiperda collected from Texas was likely controlled by a single, functionally recessive and autosomal gene. In addition, the resistance to Vip3Aa in the Texas population of S. frugiperda was not cross-resistant to Cry1F and Cry2Ab2 proteins. The interstrain complementation tests between the Texas and Louisiana resistant populations indicated the genetic basis of the resistance to Vip3Aa in these two populations are different. Considering the rapid invasion of S. frugiperda and the adoption of Vip3Aa Bt crops in the United States and other countries, results generated from this study should provide important information for management of Vip3Aa resistance in S. frugiperda worldwide.

Field efficacy of Bt cotton containing events DAS-21023-5 × DAS-24236-5 × SYN-IR102-7 against lepidopteran pests and impact on the non-target arthropod community in Brazil.

The efficacy and non-target arthropod effects of transgenic DAS-21023-5 × DAS-24236-5 × SYN-IR102-7 Bt cotton, expressing proteins Cry1Ac, Cry1F and Vip3Aa19, was examined through field trials in Brazil. Fifteen field efficacy experiments were conducted from 2014 through the 2020 growing season across six different states in Brazil to evaluate performance against key lepidopteran pests through artificial infestations of Chrysodeixis includens (Walker), Spodoptera frugiperda (J.E. Smith,1797), Spodoptera cosmioides (Walker, 1858) and Chloridea virescens (F., 1781), and natural infestations of Alabama argillacea (Hübner) and S. frugiperda. The impact of this Bt cotton technology on the non-target arthropod community in Brazilian cotton production systems was also assessed in a multi-site experiment. DAS-21023-5 × DAS-24236-5 × SYN-IR102-7 cotton significantly reduced the feeding damage caused by S. frugiperda, S. cosmioides, C. includens, C. virescens and A. argillacea, causing high levels of mortality (greater than 99%) to all target lepidopteran pests evaluated during vegetative and/or reproductive stages of crop development. Non-target arthropod community-level analyses confirmed no unintended effects on the arthropod groups monitored. These results demonstrate the value of transgenic Bt cotton containing event DAS-21023-5 × DAS-24236-5 × SYN-IR102-7 for consideration as part of an integrated approach for managing key lepidopteran pests in Brazilian cotton production systems.

Behavior and development of Tetranychus ludeni Zacher, 1913 (Acari: Tetranychidae) and physiological stress in genetically modified cotton expressing Cry1F and Cry1Ac proteins.

Genetically modified plants are one of the tactics used in integrated pest management - IPM. There is great concern about the impact of these plants on non-target organisms. On the other hand, there is little information in the literature on the effects of transgenics (Bacillus thuringiensis) Bt on populations of phytophagous mites, and the physiological responses that this attack promotes on plants. The objective of this work was to evaluate the biology of the T. ludeni mite in Bt cotton, expressing the Cry1F and Cry1Ac proteins. To evaluate the behavior of food and oviposition preference of the T. ludeni with Bt cotton and isohybrid. Verify if the physiological stress caused by T. ludeni's attack is differentiated in Bt cotton. The mites were reared in Bt cotton and isohybrid, in a total of 40 replicates in the completely randomized design and the biological cycle was evaluated. The food preference and oviposition analysis were done with 10 replicates, with choice. The physiological stress was evaluated through chlorophyll fluorescence, under greenhouse conditions. The data of the T. ludeni biology were analyzed by Student's t-test, for food and oviposition preference the chi-square test was performed. Regression models were fitted for the fluorescence parameters. The model identity test was used to evaluate the differences between Bt and isohybrid treatments. Cry1F and Cry1Ac proteins have not affected the biology of T. ludeni. The photosynthetic parameters in Bt cotton plants were less influenced by T. ludeni infestation.

Electrical signalling on Bt and non-Bt cotton plants under stress by Aphis gossypii.

Plants have developed various mechanisms to respond specifically to each biotrophic attack. It has been shown that the electrical signals emitted by plants are associated with herbivory stress responses and can lead to the activation of multiple defences. Bt cotton is a genetically modified pest-resistant plant that produces an insecticide from Bacillus thuringiensis (Bt) to control Lepidopteran species. Surprisingly, there is no study–yet, that characterizes the signalling mechanisms in transgenic cotton plants attacked by non-target insects, such as aphids. In this study, we characterized the production of electrical signals on Bt and non-Bt cotton plants infested with Aphis gossypii and, in addition, we characterized the dispersal behaviour of aphids to correlate this behaviour to plant signalling responses. Electrical signalling of the plants was recorded with an extracellular measurement technique. Impressively, our results showed that both Bt and non-Bt cotton varieties, when attacked by A. gossypii, emitted potential variation-type electrical signals and clearly showed the presence of distinct responses regarding their perception and the behaviour of aphids, with evidence of delay, in terms of signal amount, and almost twice the amount of Cry1F protein was observed on Bt cotton plants at the highest density of insects/plant. We present in our article some hypotheses that are based on plant physiology and insect behaviour to explain the responses found on Bt cotton plants under aphid stress.

Efficiency of biological control for fall armyworm resistant to the protein Cry1F.

Understanding the ecological and toxicological relationship between genetically modified cultivars (GM) and biological control agents is of great importance for discussions related to the compatability of GM cultivars and integrated management strategies for pest resistance. The present study evaluated the search behavior and predatory capacity of Orius insidiosus (Say) (Hemiptera: Anthocoridae) and Doru luteipes (Scudder) (Dermaptera: Forficulidae) on eggs and caterpillars of Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae) resistant or not to the protein Cry1F expressed in Bt corn. To determine the search time, a stopwatch was run until the capture of the first prey, predation capacity was evaluated by counting the prey remaining after 24 hours of infestation. The injuries of S. frugiperda in genetically modified and conventional corn in the presence and absence of predators was also evaluated. The predators were not able to distinguish between resistant and susceptible prey (eggs or caterpillars), given the predatory behaviour observed. There was no difference in searching time or predatory capacity between the predators for eggs and caterpillars of either resistant or susceptible S. frugiperda. In the presence of predators, the injury scores for resistant S. frugiperda on the Bt corn plants were lower. It was concluded that O. insidiosus and D. luteipes did not notice the presence of the protein Cry1F in the prey S. frugiperda, which may facilitate the combined use of GM corn and biological control in integrated management programs and for management of pest resistance.

Assessing the effects of an acute exposure to worst-case concentration of Cry proteins on zebrafish using the embryotoxicity test and proteomics analysis

Cry1C, Cry1F and Cry1Ab are insecticidal proteins from Bacillus thuringiensis (Bt) which are expressed in transgenic crops. Given the entry of these proteins into aquatic environments, it is relevant to evaluate their impacts on aquatic organisms. In this work, we sought to evaluate the effects of Cry1C, Cry1F and Cry1Ab on zebrafish embryos and larvae of a predicted worst-case scenario concentration of these proteins (set to 1.1 mg/L). For that, we coupled a traditional toxicity approach (the zebrafish embryotoxicity test and dosage of enzymatic biomarkers) to gel free proteomics analysis. At the concentration tested, these proteins did not cause adverse effects in the zebrafish early life stages, either by verifying phenotypic endpoints of toxicity or alterations in representative enzymatic biomarkers (catalase, glutathione-S-tranferase and lactate-dehydrogenase). At the molecular level, the Cry proteins tested lead to very small changes in the proteome of zebrafish larvae. In a global way, these proteins upregulated the expression of vitellogenins. Besides that, Cry1C e Cry1F deregulated heterogeneous nuclear ribonucleoproteins (Hnrnpa0l and Hnrnpaba, respectively), implicated in mRNA processing and gene regulation. Overall, these data lead to the conclusion that Cry1C, Cry1F and Cry1Ab proteins, even at a very high concentration, have limited effects in the early stages of zebrafish life.

Whole-genome sequencing to detect mutations associated with resistance to insecticides and Bt proteins in Spodoptera frugiperda.

The fall armyworm (FAW), Spodoptera frugiperda, is a major pest native to the Americas that has recently invaded the Old World. Point mutations in the target-site proteins acetylcholinesterase-1 (ace-1), voltage-gated sodium channel (VGSC) and ryanodine receptor (RyR) have been identified in S. frugiperda as major resistance mechanisms to organophosphate, pyrethroid and diamide insecticides respectively. Mutations in the adenosine triphosphate-binding cassette transporter C2 gene (ABCC2) have also been identified to confer resistance to Cry1F protein. In this study, we applied a whole-genome sequencing (WGS) approach to identify point mutations in the target-site genes in 150 FAW individuals collected from China, Malawi, Uganda and Brazil. This approach revealed three amino acid substitutions (A201S, G227A and F290V) of S. frugiperda ace-1, which are known to be associated with organophosphate resistance. The Brazilian population had all three ace-1 point mutations and the 227A allele (mean frequency = 0.54) was the most common. Populations from China, Malawi and Uganda harbored two of the three ace-1 point mutations (A201S and F290V) with the 290V allele (0.47-0.58) as the dominant allele. Point mutations in VGSC (T929I, L932F and L1014F) and RyR (I4790M and G4946E) were not detected in any of the 150 individuals. A novel 12-bp insertion mutation in exon 15 of the ABCC2 gene was identified in some of the Brazilian individuals but absent in the invasive populations. Our results not only demonstrate robustness of the WGS-based genomic approach for detection of resistance mutations, but also provide insights for improvement of resistance management tactics in S. frugiperda.

Cross-crop resistance of Spodoptera frugiperda selected on Bt maize to genetically-modified soybean expressing Cry1Ac and Cry1F proteins in Brazil

Spodoptera frugiperda is one of the main pests of maize and cotton in Brazil and has increased its occurrence on soybean. Field-evolved resistance of this species to Cry1 Bacillus thuringiensis (Bt) proteins expressed in maize has been characterized in Brazil, Argentina, Puerto Rico and southeastern U.S. Here, we conducted studies to evaluate the survival and development of S. frugiperda strains that are susceptible, selected for resistance to Bt-maize single (Cry1F) or pyramided (Cry1F/Cry1A.105/Cry2Ab2) events and F1 hybrids of the selected and susceptible strains (heterozygotes) on DAS-444Ø6-6 × DAS-81419-2 soybean with tolerance to 2,4-d, glyphosate and ammonium glufosinate herbicides (event DAS-444Ø6-6) and insect-resistant due to expression of Cry1Ac and Cry1F Bt proteins (event DAS-81419-2). Susceptible insects of S. frugiperda did not survive on Cry1Ac/Cry1F-soybean. However, homozygous-resistant and heterozygous insects were able to survive and emerge as fertile adults when fed on Cry1Ac/Cry1F-soybean, suggesting that the resistance is partially recessive. Life history studies revealed that homozygous-resistant insects had similar development, reproductive performance, net reproductive rate, intrinsic and finite rates of population increase on Cry1Ac/Cry1F-soybean and non-Bt soybean. In contrast, heterozygotes had their fertility life table parameters significantly reduced on Cry1Ac/Cry1F-soybean. Therefore, the selection of S. frugiperda for resistance to single and pyramided Bt maize can result in cross-crop resistance to DAS-444Ø6-6 × DAS-81419-2 soybean. The importance of these results to integrated pest management (IPM) and insect resistance management (IRM) programs is discussed.

Efficacy of Bacillus thuringiensis (Bt) maize expressing Cry1F, Cry1A.105, Cry2Ab2 and Vip3Aa20 proteins to manage the fall armyworm (Lepidoptera: Noctuidae) in Brazil

Recommendations to manage the impact of Spodoptera frugiperda (Lepidoptera: Noctuidae) to maize production include the use of Bt crops where approved, combined with the adoption of integrated pest management (IPM) and insect resistance management (IRM) practices. This study evaluated the efficacy of Bt maize expressing proteins Cry1F + Cry 1A.105 + Cry2Ab2 + Vip3Aa20 to control S. frugiperda under field conditions in Brazil. This Bt maize technology effectively managed seedling cutting injury, significantly reduced foliar feeding and achieved near 100% protection from kernel-feeding caused by both a Cry1F-resistant strain and a field-collected population of S. frugiperda. Consistent field-collected data across several years and planting regions suggest that this Bt maize technology will be an important tool for the effective management of all important feeding injury types caused by S. frugiperda in Brazil. As with any Bt maize hybrid, its implementation within IPM and IRM practices should help growers maintain maize crop sustainability in Brazil.

Fluorescence of chlorophyll a in transgenic maize with herbicide application and attacked by Spodoptera frugiperda (Lepidoptera: Noctuidae)

Physiological changes in maize plants, submitted to herbicides and pests, should be studied for integrated management in this crop. The objective was to evaluate the chlorophyll a fluorescence in maize genotypes damaged or not by Spodoptera frugiperda, and sprayed with herbicides. The herbicides atrazine and nicosulfuron were applied to the plants at doses of 3 kg ha− 1 and 0.06 kg ha− 1, respectively. A randomized block design in a factorial scheme with an additional treatment 3 (2 × 2 + 1) was used, being the factor A: maize leaves exposed or not to the attack of S. frugiperda; factor B: Herculex® (transgenic maize encoding the Cry1F protein) and Isohybrid (non-transgenic maize) genotypes and an additional treatment represented by the PowerCore® genotype (stacked transgenic maize encoding the Cry1F, Cry1A.105 and Cry2Ab2 proteins). The initial fluorescence of chlorophyll a (F0 - electrons quantum− 1), maximum fluorescence (Fm - electron quantum− 1), ratio between variable fluorescence and maximum fluorescence of chlorophyll a (Fv/Fm) and electron transport rate ETR-µmols electrons m− 2 s− 1) were analyzed. The initial fluorescence of chlorophyll a increased and Fm, Fv/Fm and ETR decreased in plants attacked by the pest. The PowerCore® chlorophyll fluorescence remained in characteristic patterns. Chlorophyll a fluorescence was similar between genotypes with or without herbicides application. Damage by S. frugiperda increased the initial fluorescence of chlorophyll a and reduced the variable fluorescence/maximum fluorescence ratio and electron transport rate, indicating physiological stress in maize plants. The herbicides do not cause physiological stress in plants of maize genotypes with characteristic values of chlorophyll a fluorescence.

Agronomic parameters and morpho-agronomic characteristics of genetically modified maize hybrids compared to conventional maize hybrids

The objectives were to study the behavior of fifteen pre-commercial upland maize hybrids, analyze their agronomic performance regarding grain yield, and evaluate productivity components, as well as morpho-agronomic characteristics, in the Midwest Region of Brazil. Two experiments were conducted in the municipalities of Formosa-GO and Planaltina-DF, 2016/17 crop year. Both consisted of five pre-commercial maize hybrid platforms (HPA252, HPB262, HPB621, HPB646, and HPD354). Each platform consisted of three different versions: conventional, transgenic with a Bt gene that expresses the protein Cry1F, and transgenic with two Bt genes that express the proteins Cry1F and Cry1AB. The experiment was randomized blocks with four replications. The experimental plot was four lines five meters long considering the two central lines as useful. The lines were spaced 0.75 meters apart, and the final density was five plants per linear meter. To estimate grain yield, the plots were harvested, and the weight was extrapolated to kg.ha-1. The moisture was standardized at 14%. Data were submitted to analysis of variance, and the means were compared by Tukey test at 5% probability using the Sisvar software. Grain yield between treatments ranged from 8,381 to 12,908 kg.ha-1, and the average yield was 11,234 kg.ha-1. The parameters evaluated were divided into two groups. The first group contained parameters determining grain yield: grain depth, thousand grains weight, number of rows of grains, number of grains per row, and grain yield. The second group contained morpho-agronomic parameters that directly interfere with resistance to lodging and plant breaking: plant height, ear insertion height, and mean stem diameter. There was no direct effect of the transgenes on the evaluated hybrids since the classification of productivity was not divided into conventional and transgenic classes. It is suggested that the HPA252YH, HP621H and HP646H versions be discarded because grain yield performance was unsatisfactory compared to their respective conventional and transgenic counterparts. The parameters GD and W1000 are more effective for grain yield estimation than NR and NG. It was observed for HP621H that, in addition to a lower grain yield, there was a significant reduction in stem diameter, indicating a possible reduction in lodging tolerance and/or plant breaking when exposed to adverse climatic conditions such as windstorms. For the HPD354H version, the release of the transgenic counterpart HPD354YH is preferentially indicated since the H version had a significant reduction in stem diameter and a possible greater tolerance to lodging and/or plant breaking under the same conditions.