Analiza P. Alves

Fitness costs of Cry1F resistance in fall armyworm, Spodoptera frugiperda

Transgenic corn, Zea mays L., expressing the Bacillus thuringiensis Berliner (Bt) protein Cry1F has been registered for Spodoptera frugiperda (J. E. Smith) control since 2003 in the USA. Unexpected damage to Cry1F corn was reported in 2006 in Puerto Rico, and Cry1F resistance in S. frugiperda from Puerto Rico was documented. The study of fitness costs associated with insect resistance to Bt insecticidal proteins is important for understanding resistance evolution and for evaluating resistance management practices used to mitigate resistance to transgenic corn. Currently, no studies have addressed the fitness costs associated with Cry1F resistance in S. frugiperda. In this study, susceptible and resistant strains with similar genetic background and their reciprocal crosses were used to estimate Cry1F resistance fitness costs. Comparisons between life‐history traits and population growth rates of homozygous susceptible, heterozygous and homozygous resistant S. frugiperda were used to determine whether the resistance is associated with fitness costs. Major fitness costs were not apparent in either heterozygotes or homozygous resistant insects. However, there was a slight indication of hybrid vigour in the heterozygotes. Additionally, two lines in which the frequency of the resistant alleles was fixed at 0.5 were followed for seven generations, after which the frequency of resistant alleles slightly decreased in both lines. The lack of strong fitness costs associated with Cry1F resistance in S. frugiperda indicates that initial allele frequencies may be higher than expected in field populations and will tend to remain stable in field populations in the absence of selection pressure (e.g. Puerto Rico).

Applying an Integrated Refuge to Manage Western Corn Rootworm (Coleoptera: Chrysomelidae): Effects on Survival, Fitness, and Selection Pressure

ABSTRACT The refuge strategy can delay resistance of insect pests to transgenic maize producing toxins from Bacillus thuringiensis (Bt). This is important for the western corn rootworm, Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae), because of its history of adaptation to several management practices. A 2-yr study across four locations was conducted to measure the effects of integrated refuge (i.e., blended refuge) on western corn rootworm survival to adulthood, fitness characteristics, and susceptibility to Bt maize in the subsequent generation. The treatments tested in this study were as follows: a pure stand of Bt maize (event DAS-59122-7, which produces Bt toxins Cry34Ab1/Cry35Ab1), a pure stand of refuge (non-Bt maize), and two variations on an integrated refuge consisting of 94.4% Bt maize and 5.6% non-Bt maize. Within the two integrated refuge treatments, refuge seeds received a neonicotinoid insecticidal seed treatment of either 1.25 mg clothianidin per kernel or 0.25 mg thiamethoxam per kernel. Insects in the pure stand refuge treatment had greater survival to adulthood and earlier emergence than in all other treatments. Although fecundity, longevity, and head capsule width were reduced in treatments containing Bt maize for some site by year combinations, Bt maize did not have a significant effect on these factors when testing data across all sites and years. We found no differences in susceptibility of larval progeny to Bt maize in bioassays using progeny of adults collected from the four treatments.

Fitness costs and stability of Cry1Fa resistance in Brazilian populations of Spodoptera frugiperda

BACKGROUND The presence of fitness costs of resistance to Bacillus thuringiensis (Bt) insecticidal proteins in insect populations may delay or even reverse the local selection of insect resistance to Bt transgenic crops, and deserves rigorous investigation. Here we assessed the fitness costs associated with Cry1Fa resistance in two strains of fall armyworm, Spodoptera frugiperda (Lepidoptera: Noctuidae), derived from field collections in different Brazilian regions and further selected in the laboratory for high levels of resistance to Cry1Fa using leaves of TC1507 corn. RESULTS Fitness components were compared using paired resistant and susceptible strains with similar genetic backgrounds and F1 generations from reciprocal crosses, all of them reared on non-transgenic corn leaves. No apparent life history costs in the larval stage were observed in the Bt-resistant strains. Moreover, the resistance remained stable for seven generations in the absence of selection, with no decrease in the proportion of resistant individuals. Larval respiration rates were also similar between resistant and susceptible homozygotes, and heterozygotes displayed respiration rates and demographic performance equal or superior to those of susceptible homozygotes. CONCLUSION In combination, these results indicate the lack of strong fitness costs associated with resistance to Cry1Fa in the fall armyworm strains studied. These findings suggest that Cry1Fa resistance in S. frugiperda populations is unlikely to be counterselected in Cry1Fa-free environments.

Effect of Cry1F maize on the behavior of susceptible and resistant Spodoptera frugiperda and Ostrinia nubilalis

Understanding the behavior of pests targeted with Bacillus thuringiensis Berliner (Bt) crops is important to define resistance management strategies. Particularly the study of larval movement between plants is important to determine the feasibility of refuge configurations. Exposure to Bt maize, Zea mays L. (Poaceae), has been suggested to increase larval movement in lepidopteran species but few studies have examined the potential for resistance to interact with behavioral responses to Bt toxins. Choice and no‐choice experiments were conducted with Spodoptera frugiperda (JE Smith) (Lepidoptera: Noctuidae) and Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae) to determine whether Cry1F resistance influences neonate movement. Leaf discs of Cry1F maize and the corresponding isoline were used to characterize behavioral responses. In both experiments, the location (on or off of plant tissues) and mortality of susceptible and Cry1F resistant neonates was recorded for 5 days, but the analysis of larvae location was performed until 7 h. Our results indicated no strong difference between resistant and susceptible phenotypes in S. frugiperda and O. nubilalis, although a small percentage of susceptible neonates in both species abandoned maize tissue expressing Cry1F. However, significant behavioral differences were observed between species. Ostrinia nubilalis exhibited increased movement between leaf discs, whereas S. frugiperda selected plant tissue within the first 30 min and remained on the chosen plant regardless of the presence of Cry1F. Spodoptera frugiperda reduced larval movement may have implications to refuge configuration. This study represents the first step toward understanding the effects of Cry1F resistance on Lepidoptera larval behavior. Information regarding behavioral differences between species could aid in developing better and more flexible resistance management strategies.

Survival of Corn Earworm (Lepidoptera: Noctuidae) on Bt Maize and Cross-Pollinated Refuge Ears From Seed Blends

Abstract Refuge is mandated in the United States where genetically modified maize (Zea mays L.) expressing insecticidal proteins derived from Bacillus thuringiensis Berliner (Bt) are cultivated. Currently, refuge is deployed in different ways including blocks, field strips, or seed blends containing Bt and non-Bt maize. Seed blends provide practical advantages for refuge implementation. However, concerns related to the movement of insect larvae, potential differential survival of heterozygous resistant larvae, reduction in insect production, and cross-pollination of ears resulting in sublethal selection, have delayed seed blend use for Lepidoptera in the southern United States, where maize plantings are used as refuge for Helicoverpa zea (Boddie). In this study, we evaluated the relative survival of H. zea in Bt events and in seed blends compared with pure stand refuge and the relative survival of H. zea on the individual components of the pyramid 1507xMON810xMIR162. The results showed variation on the production of H. zea in refuge plants from seed blends compared with pure stand refuge plants. The relative survival of H. zea on the events 1507, MON810, MIR162, and 1507xMON810xMIR162 ranked similarly across the three locations tested. These results can be used in computer simulation modeling efforts to evaluate the feasibility of seed blends as a refuge deployment strategy with the pyramid 1507xMON810xMIR162. Because the reduction on survival of H. zea due to blending was variable, a sensitivity analysis that includes all possible scenarios of reduction in survival should be considered.

Characterization of field-evolved resistance to Bacillus thuringiensis-derived Cry1F δ-endotoxin in Spodoptera frugiperda populations from Argentina

BACKGROUND Transgenic maize (Zea mays L.) event TC1507 (Herculex® I insect protection), expressing Cry1F δ-endotoxin derived from Bacillus thuringiensis var. aizawai, was commercialized in 2003 in the Americas. Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) susceptibility to Cry1F was monitored annually across several regions in Argentina using diagnostic concentration bioassays. Reduced performance of TC1507 maize against S. frugiperda was reported in 2013. A resistant population was established in the laboratory and the dominance of Cry1F resistance was characterized. RESULTS During 2012-2015, high-survivorship of several populations was observed in the resistance monitoring program. Reciprocal crosses of a Cry1F-resistant population with a Cry1F-susceptible population were evaluated to calculate effective dominance (DML ) based on mortality levels observed at 100 µg/ml Cry1F. Two additional dominance levels (DLC and DEC ) were calculated using lethal (LC50 ) or effective concentration (EC50 ) derived from concentration-response bioassays. Estimates indicated that Cry1F resistance in S. frugiperda in Argentina was either highly recessive (DML = 0.005) or incompletely recessive (DLC < 0.26 and DEC < 0.19). CONCLUSION This study is the first documented confirmation and characterization of S. frugiperda Cry1F field-evolved resistance in Argentina. The resistance to Cry1F in S. frugiperda populations collected in Argentina, is autosomal and incompletely recessive similar to the resistance reported in Brazil.

Magnitude and Allele Frequency of Cry1F Resistance in Field Populations of the Fall Armyworm (Lepidoptera: Noctuidae) in Brazil

Abstract Bacillus thuringiensis (Bt) corn producing the Cry1F protein was the first highly efficacious Bt corn deployed against the fall armyworm Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) in Brazil, but reduced efficacy of this technology against the fall armyworm has been reported in some regions of the country. Here, we surveyed Cry1F resistance allele frequency and susceptibility of eight S. frugiperda populations collected in 2013 from non-Bt fields in different regions of Brazil. In F1 screen experiments, the overall frequency of the Cry1F resistance alleles in Brazilian populations was estimated at 0.24, with 95% credibility interval between 0.18 and 0.25. In concentration–response bioassays, five of the eight populations surveyed exhibited significant resistance levels, which were over 32 times higher than that of the standard susceptible laboratory strain. The estimates of Cry1F resistance allele frequency were positively correlated with those of median effective or lethal concentrations (i.e., EC50 or LC50). These results show that the allelic frequency and the magnitude of Cry1F resistance are high in field populations of S. frugiperda in Brazil, indicating a challenging situation for resistance management.

Plant Incorporated Protectants and Insect Resistance

The advancements of recombinant DNA and plant transformation technologies have changed the face of commercial agriculture for insect control by literally transforming insect control agents into plant incorporated protectants (PIPs). In planta delivery has important advantages over conventional foliar or soil-applied alternatives, including reduced environmental footprint, selective targeting of host pest insects, and simplified season-long delivery of protection. However, the potential for insect resistance that has limited the durability of conventional exogenous pesticides remains a challenge for PIPs. The long-term sustainability of transgenic insect control technologies will depend on the implementation of responsible integrated pest management (IPM) tactics as well as the continued discovery of new modes of action. As will be discussed in this chapter, the insecticidal proteins from Bacillus thuringiensis (Bt) have served to validate both the feasibility and value of the transgenic approach. The variety of Bt insecticidal proteins that continue to be discovered as well as the increased understanding of their mode of action should support their use well into the future, but alternatives to Bt with novel of modes of action would be highly beneficial as insect resistance management (IRM) tools. Some of these alternatives will be discussed as well.