Oviposition patterns of primary lepidopteran defoliators in soybean and the impact on structured refuge recommendations.

BACKGROUND: Transgenic Bt technology in soybean, with plants expressing Cry1Ac, has been adopted as an insect pest management tool. It was first adopted in large areas of South America and Asia in 2013. The risk of resistance in target pests to this technology demands insect resistance management (IRM) programs. In Brazil, a structured refuge (area of non-Bt soybean) planted adjacent to the Bt soybean crop has been an important IRM recommendation, particularly for the primary lepidopteran defoliators Anticarsia gemmatalis (Lepidoptera: Erebidae) and Chrysodeixis includens (Lepidoptera: Noctuidae). The overall goal of this study was to validate IRM recommendations to Bt soybean. The objectives were to document the impact of soybean phenology, cultivar choice and non-Bt soybean defoliation on moth oviposition. In addition, a mark-release-recapture study estimated the dispersal capacity of these species. Five field experiments per species were performed for 3 years. RESULTS: Our results revealed an increase in A. gemmatalis and C. includens oviposition, respectively, on Bt plants as a consequence of the difference in plant growth stage at the time of oviposition. Defoliation of non-Bt plants significantly increased the oviposition preference of both moth species for Bt plants. The mark-release-recapture experiment indicated an average dispersal distance of ~300 m from the release point for A. gemmatalis, with maximum recapture at 1000 m. CONCLUSION: Overall, our findings emphasize the importance of planting synchronization of Bt soybean and the structured refuge. In addition, when operational aspects in large soybean areas challenge this recommendation, the priority should be for planting the refuge area first. This approach will minimize the impact of selective oviposition of A. gemmatalis and C. includens. © 2024 Society of Chemical Industry.

The Efficacy of Entomopathogenic Nematodes Plus an Adjuvant against Helicoverpa zea and Chrysodeixis includens in Aboveground Applications.

In the southern United States, corn earworm, Helicoverpa zea (Boddie), and soybean looper, Chrysodeixis includens (Walker) are economically important crop pests. Although Bt crops initially provided effective control of target pests such as H. zea, many insect pests have developed resistance to these Bt crops. Alternative approaches are needed, including biological control agents such as entomopathogenic nematodes (EPNs). However, the effectiveness of EPNs for aboveground applications can be limited due to issues such as desiccation and ultraviolet radiation. Effective adjuvants are needed to overcome these problems. Ten strains of EPNs were tested for virulence against eggs, first to fourth instars, fifth instars, and pupae of H. zea and C. includens in the laboratory. These 10 EPN strains were Heterorhabditis bacteriophora (HP88 and VS strains), H. floridensis (K22 strain), Hgkesha (Kesha strain), Steinernema carpocapsae (All and Cxrd strains), S. feltiae (SN strain), S. rarum (17c+e strain), and S. riobrave (355 and 7-12 strains). EPNs could infect eggs of H. zea or C. includens in the laboratory, but the infection was low. The mortality caused by 10 EPN strains in seven days was significantly higher for the first to fourth instars of H. zea compared to the control, as was the fifth instars of H. zea. Similarly, for the first to fourth and fifth instars of C. includens, the mortality was significantly higher compared to the controls, respectively. However, only S. riobrave (355) had significantly higher mortality than the control for the pupae of H. zea. For the pupae of C. includens, except for H. bacteriophora (HP88), S. rarum (17c+e), and H. floridensis (K22), the mortality of the other seven strains was significantly higher than the control. Subsequently, S. carpocapsae (All) and S. riobrave (7-12) were chosen for efficacy testing in the field with an adjuvant 0.066% Southern Ag Surfactant (SAg Surfactant). In field experiments, the SAg Surfactant treatment significantly increased the mortality and EPN infection for S. carpocapsae (All) on first instars of H. zea in corn plant whorls. On soybean plants, with the SAg Surfactant, S. carpocapsae (All) was more effective than S. riobrave (7-12) on fifth instars of C. includens. This study indicates that EPNs can control H. zea and C. includens, and SAg Surfactant can enhance EPN efficacy.

Molecular characterization and expression patterns of a ryanodine receptor in soybean looper, Chrysodeixis includens.

Diamide insecticides, such as chlorantraniliprole, have been widely used to control insect pests by targeting the insect ryanodine receptor (RyR). Due to the efficacious insecticidal activity of diamides, as well as an increasing number of resistance cases, the molecular structure of RyR has been studied in many economically important insects. However, no research has been conducted on diamide resistance and RyR in the soybean looper, Chrysodeixis includens, a significant crop pest. In this study, we found moderate resistance to chlorantraniliprole in a field population from Puerto Rico and sequenced the full-length cDNA of the C. includens RyR gene, which encodes a 5124 amino acid-long protein. Genomic analysis revealed that the CincRyR gene consists of 113 exons, one of the largest exon numbers reported for RyR. Alternative splicing sites were detected in the cytosolic region. The protein sequence showed high similarity to other noctuid RyRs. Conserved structural features included the selectivity filter motif critical for ryanodine binding and ion conduction, as well as various domains involved in ion transport. Two mutation sites associated with diamide resistance in other insects were screened but not found in the Puerto Rico field populations or in the susceptible lab strain. Gene expression analysis indicated high expression of RyR in the third instar larval stage, particularly in muscle-containing tissues. Furthermore, exposure to a sublethal dose of chlorantraniliprole reduced RyR expression levels after 96 h. This study provides a molecular basis for understanding RyR structure and sheds light on potential mechanisms of diamide resistance in C. includens.

Regional pest suppression associated with adoption of Cry1Ac soybean benefits pest management in tropical agriculture.

BACKGROUND: Bacillus thuringiensis (Bt) crops have been adopted worldwide, providing high-level protection from insect pests. Furthermore, Bt crops preserve natural enemies, promote higher yield, and economically benefit farmers. Although regional pest suppression by widespread Bt crop adoption has been observed in temperate regions, this possibility remains uncertain in tropical areas due to the high diversity of alternative hosts and mild winters. RESULTS: Evidence of regional reduction in insecticide use across areas was observed in Brazil where Cry1Ac soybean has been grown since 2013, with up to 50% reduction in the number of insecticide sprays for managing lepidopteran pests on non-Bt soybean observed at specific locations from 2012 to 2019. Pest monitoring data from four mesoregions across 5 years of commercial plantings of Cry1Ac soybean from December 2014 to July 2019 showed reduced numbers of Chrysodeixis includens moths captured in pheromone traps across years at all locations. The number of Helicoverpa spp. moths captured also was reduced at three locations. CONCLUSION: We provide evidence for regional suppression of lepidopteran pests and reduced insecticide use with the widespread adoption of Cry1Ac soybean in Brazil, bringing economic, social and environmental benefits. © 2022 Society of Chemical Industry.

Generation of Variability in Chrysodeixis includens Nucleopolyhedrovirus (ChinNPV): The Role of a Single Variant.

The mechanisms generating variability in viruses are diverse. Variability allows baculoviruses to evolve with their host and with changes in their environment. We examined the role of one genetic variant of Chrysodeixis includens nucleopolyhedrovirus (ChinNPV) and its contribution to the variability of the virus under laboratory conditions. A mixture of natural isolates (ChinNPV-Mex1) contained two genetic variants that dominated over other variants in individual larvae that consumed high (ChinNPV-K) and low (ChinNPV-E) concentrations of inoculum. Studies on the ChinNPV-K variant indicated that it was capable of generating novel variation in a concentration-dependent manner. In cell culture, cells inoculated with high concentrations of ChinNPV-K produced OBs with the ChinNPV-K REN profile, whereas a high diversity of ChinNPV variants was recovered following plaque purification of low concentrations of ChinNPV-K virion inoculum. Interestingly, the ChinNPV-K variant could not be recovered from plaques derived from low concentration inocula originating from budded virions or occlusion-derived virions of ChinNPV-K. Genome sequencing revealed marked differences between ChinNPV-K and ChinNPV-E, with high variation in the ChinNPV-K genome, mostly due to single nucleotide polymorphisms. We conclude that ChinNPV-K is an unstable genetic variant that is responsible for generating much of the detected variability in the natural ChinNPV isolates used in this study.

Metabolomics Differences of Glycine max QTLs Resistant to Soybean Looper.

Quantitative trait loci (QTLs) E and M are major soybean alleles that confer resistance to leaf-chewing insects, and are particularly effective in combination. Flavonoids and/or isoflavonoids are classes of plant secondary metabolites that previous studies agree are the causative agents of resistance of these QTLs. However, all previous studies have compared soybean genotypes that are of dissimilar genetic backgrounds, leaving it questionable what metabolites are a result of the QTL rather than the genetic background. Here, we conducted a non-targeted mass spectrometry approach without liquid chromatography to identify differences in metabolite levels among QTLs E, M, and both (EM) that were introgressed into the background of the susceptible variety Benning. Our results found that E and M mainly confer low-level, global differences in distinct sets of metabolites. The isoflavonoid daidzein was the only metabolite that demonstrated major increases, specifically in insect-treated M and EM. Interestingly, M confers increased daidzein levels in response to insect, whereas E restores M's depleted daidzein levels in the absence of insect. Since daidzein levels do not parallel levels of resistance, our data suggest a novel mechanism that the QTLs confer resistance to insects by mediating changes in hundreds of metabolites, which would be difficult for the insect to evolve tolerance. Collective global metabolite differences conferred by E and M might explain the increased resistance of EM.

Temporal Occurrence of Plusiinae on Soybean in the Mississippi River Delta.

The subfamily Plusiinae of the moth family Noctuidae is made up 400 species worldwide. Two species of the subfamily, the soybean looper, Chrysodeixis includens (Walker), and the cabbage looper, Trichoplusia ni (Hübner) (Lepidoptera: Noctuidae), are important defoliating insect pests of various field crops and have been the subject of previous Plusiinae surveys in the Southern U.S. Soybean fields were sampled in the Mississippi Delta from 2010 to 2012 to determine the temporal occurrence of various Plusiinae species on soybean. As in previous surveys, C. includens was the most common Plusiinae species in soybean during the 3-yr survey, especially in late season collections (July-September). Rachiplusia ou (Guenée) (Lepidoptera: Noctuidae) was the predominant species observed in early season collections (May-early July). Populations of R. ou during the first sample dates during 2010 were much higher than those observed during the other years of the survey. Only three collected larvae successfully developed into T. ni adults, one each collected during May, June, and July. Although R. ou was not commonly reported in previous studies on soybean, it occurred in low numbers during June and July during the 3-yr study. The temporal occurrence and species composition followed a predictable pattern in all 3 yr of the study.

Elucidating Efficacy of Ingested Positively Charged Zein Nanoparticles Against Noctuidae.

A meridic diet overlay bioassay using empty, positively charged zein nanoparticles ((+)ZNP) was performed on soybean looper (Chrysodeixis includens (Walker)), tobacco budworm (Heliothis virescens (F.)), and velvetbean caterpillar (Anticarsia gemmatalis Hübner) (Lepidoptera: Noctuidae). Assessment of effects on mortality and development weights 7 d after ingestion of (+)ZNP were evaluated on larvae of each species. Treatments involved different concentrations, with H. virescens and A. gemmatalis offered 0 and 3,800 ppm (+)ZNP, whereas C. includens colonies were offered 0, 630, 1,260, and 2,520 ppm (+)ZNP. Mortality of A. gemmatalis and C. includens increased after ingestion of the highest (+)ZNP concentrations, while H. virescens neonate mortality was unaffected. Neonate and third-instar weights of A. gemmatalis and C. includens, and neonate H. virescens, decreased with high (+)ZNP concentrations. Following mortality results from A. gemmatalis neonates, a concentration response test was performed using a range of (+)ZNP concentrations. The LC50 for A. gemmatalis was 1,478 ppm. The potential of (+)ZNP as a pest management tactic is discussed.

A Novel Alphabaculovirus from the Soybean Looper, Chrysodeixis includens, that Produces Tetrahedral Occlusion Bodies and Encodes Two Copies of he65.

Isolates of the alphabaculovirus species, Chrysodeixis includens nucleopolyhedrovirus, have been identified that produce polyhedral occlusion bodies and infect larvae of the soybean looper, Chrysodeixis includens. In this study, we report the discovery and characterization of a novel C. includens-infecting alphabaculovirus, Chrysodeixis includens nucleopolyhedrovirus #1 (ChinNPV#1), that produces tetrahedral occlusion bodies. In bioassays against C. includens larvae, ChinNPV #1 exhibited a degree of pathogenicity that was similar to that of other ChinNPV isolates, but killed larvae more slowly. The host range of ChinNPV#1 was found to be very narrow, with no indication of infection occurring in larvae of Trichoplusia ni and six other noctuid species. The ChinNPV#1 genome sequence was determined to be 130,540 bp, with 126 open reading frames (ORFs) annotated but containing no homologous repeat (hr) regions. Phylogenetic analysis placed ChinNPV#1 in a clade with other Group II alphabaculoviruses from hosts of lepidopteran subfamily Plusiinae, including Chrysodeixis chalcites nucleopolyhedrovirus and Trichoplusia ni single nucleopolyhedrovirus. A unique feature of the ChinNPV#1 genome was the presence of two full-length copies of the he65 ORF. The results indicate that ChinNPV#1 is related to, but distinct from, other ChinNPV isolates.

Owlet moths (Lepidoptera: Noctuoidea) associated with Bt and non- Bt soybean in the brazilian savanna / Noctuóides (Lepidoptera: Noctuoidea) associados a soja Bt e não-Bt no Cerrado brasileiro

Abstract The use of GMO expressing Bt toxin in soybean production has increased significantly in the last years in Brazil in order to manage the damage caused by lepidopteran pests. In this study, we compared the richness and abundance of owlet moths (Noctuoidea) associated with Bt and non-Bt soybean. We determined the temporal variations as a function of phenology, and correlated the population variations of the most common species with meteorological variables. The research was conducted at the experimental area of Embrapa Cerrados. The collection method used was differentiated being suppressive and absolute. A total of 13 species were collected, of which eight occurred on Bt soybeans. The most representative taxa were Chrysodeixis includens (72.87%), Anticarsia gemmatalis (18.17%) and Spodoptera spp (5.22%). The number of larvae belonging to species targeted by the Bt technology was 10 times lower on Bt than on non-Bt soybeans. Utetheisa ornatrix and Elaphria deltoides were recorded on soybean for the first time, observing larvae of both species in non-Bt soybean and those of U. ornatrix also in Bt soybean. Only A. gemmatalis larvae correlated (p <0.05) negatively with precipitation. This study provided field information on the abundance and species richness of owlet moths on non-Bt soybeans, associated with the effects of Bt soybean. When considering the different levels of infestation between cultivars as a criterion, larvae monitoring is of substantial importance in order to develop the lost control program.

Resumo O uso de OGM que expressam toxina Bt na produção de soja tem aumentado significativamente nos últimos anos no Brasil e são utilizados para conter os danos causados ​​pelos lepidópteros pragas. Neste estudo comparamos a riqueza e a abundância de Noctuoides (Noctuoidea) associados à soja Bt e não-Bt. Determinamos as variações temporais em função da fenologia e correlacionamos às variações populacionais das espécies mais comuns com variáveis ​​meteorológicas. A pesquisa foi conduzida na área experimental da Embrapa Cerrados. O método de coleta utilizado foi diferenciado sendo supressivo e absoluto. Um total de 13 espécies foram coletadas, das quais oito ocorreram em soja Bt. Os taxa mais representativos foram Chrysodeixis includens, Anticarsia gemmatalis e Spodoptera spp. O número de larvas pertencentes às espécies alvo da tecnologia Bt foram 10 vezes menores na soja Bt do que em soja não-Bt . Utetheisa ornatrix e Elaphria deltoides foram registradas na soja pela primeira vez, observando-se larvas de ambas espécies na soja não-Bt e as de U. ornatrix também na soja Bt. Somente as larvas de A. gemmatalis se correlacionaram (p <0,05) negativamente com a precipitação. Este estudo forneceu informações em campo sobre a abundância e riqueza de espécies na soja não- Bt, associada aos efeitos da soja Bt. A importância do monitoramento das lagartas é substancial, a fim de tomar a melhor decisão de controle, considerando-se os diferentes níveis de infestação entre cultivares como critério.

Domain III of Cry1Ac Is Critical to Binding and Toxicity against Soybean Looper (Chrysodeixis includens) but Not to Velvetbean Caterpillar (Anticarsia gemmatalis).

Insecticidal proteins Cry1Ac and Cry2Ac7 from the bacterium Bacillus thuringiensis (Bt) belong to the three-domain family of Bt toxins. Commercial transgenic soybean hybrids produce Cry1Ac to control the larvae of the soybean looper (Chrysodeixis includens) and the velvet bean caterpillar (Anticarsia gemmatalis). The specificity of Cry1Ac is determined by loops extending from domain II and regions of domain III in the three-dimensional structure of the toxin. In this study, we constructed a hybrid toxin (H1.2Ac) containing domains I and II of Cry1Ac and domain III of Cry2Ac7, in an attempt to obtain a protein with enhanced toxicity compared to parental toxins. Bioassays with H1.2Ac revealed toxicity against the larvae of A. gemmatalis but not against C. includens. Saturation binding assays with radiolabeled toxins and midgut brush border membrane vesicles demonstrated no specific H1.2Ac binding to C. includens, while binding in A. gemmatalis was specific and saturable. Results from competition binding assays supported the finding that Cry1Ac specificity against A. gemmatalis is mainly dictated by domain II. Taken together, these distinct interactions with binding sites may help explain the differential susceptibility to Cry1Ac in C. includens and A. gemmatalis, and guide the design of improved toxins against soybean pests.

Owlet moths (Lepidoptera: Noctuoidea) associated with Bt and non- Bt soybean in the brazilian savanna

Abstract The use of GMO expressing Bt toxin in soybean production has increased significantly in the last years in Brazil in order to manage the damage caused by lepidopteran pests. In this study, we compared the richness and abundance of owlet moths (Noctuoidea) associated with Bt and non-Bt soybean. We determined the temporal variations as a function of phenology, and correlated the population variations of the most common species with meteorological variables. The research was conducted at the experimental area of Embrapa Cerrados. The collection method used was differentiated being suppressive and absolute. A total of 13 species were collected, of which eight occurred on Bt soybeans. The most representative taxa were Chrysodeixis includens (72.87%), Anticarsia gemmatalis (18.17%) and Spodoptera spp (5.22%). The number of larvae belonging to species targeted by the Bt technology was 10 times lower on Bt than on non-Bt soybeans. Utetheisa ornatrix and Elaphria deltoides were recorded on soybean for the first time, observing larvae of both species in non-Bt soybean and those of U. ornatrix also in Bt soybean. Only A. gemmatalis larvae correlated (p 0.05) negatively with precipitation. This study provided field information on the abundance and species richness of owlet moths on non-Bt soybeans, associated with the effects of Bt soybean. When considering the different levels of infestation between cultivars as a criterion, larvae monitoring is of substantial importance in order to develop the lost control program.

Resumo O uso de OGM que expressam toxina Bt na produção de soja tem aumentado significativamente nos últimos anos no Brasil e são utilizados para conter os danos causados pelos lepidópteros pragas. Neste estudo comparamos a riqueza e a abundância de Noctuoides (Noctuoidea) associados à soja Bt e não-Bt. Determinamos as variações temporais em função da fenologia e correlacionamos às variações populacionais das espécies mais comuns com variáveis meteorológicas. A pesquisa foi conduzida na área experimental da Embrapa Cerrados. O método de coleta utilizado foi diferenciado sendo supressivo e absoluto. Um total de 13 espécies foram coletadas, das quais oito ocorreram em soja Bt. Os taxa mais representativos foram Chrysodeixis includens, Anticarsia gemmatalis e Spodoptera spp. O número de larvas pertencentes às espécies alvo da tecnologia Bt foram 10 vezes menores na soja Bt do que em soja não-Bt . Utetheisa ornatrix e Elaphria deltoides foram registradas na soja pela primeira vez, observando-se larvas de ambas espécies na soja não-Bt e as de U. ornatrix também na soja Bt. Somente as larvas de A. gemmatalis se correlacionaram (p 0,05) negativamente com a precipitação. Este estudo forneceu informações em campo sobre a abundância e riqueza de espécies na soja não- Bt, associada aos efeitos da soja Bt. A importância do monitoramento das lagartas é substancial, a fim de tomar a melhor decisão de controle, considerando-se os diferentes níveis de infestação entre cultivares como critério.

Efficacy of Soybean's Event DAS-81419-2 Expressing Cry1F and Cry1Ac to Manage Key Tropical Lepidopteran Pests Under Field Conditions in Brazil.

Bacillus thuringiensis (Bt) event DAS-81419-2 (Conkesta technology) in soybean, Glycine max (L.) Merrill, expresses Cry1F and Cry1Ac proteins to provide protection from feeding by several lepidopteran pests. A total of 27 field experiments across nine locations were conducted from 2011 to 2015 in southern and central Brazil to characterize the efficacy of DAS-81419-2 soybean infested with Anticarsia gemmatalis Hübner (Lepidoptera: Erebidae), Chrysodeixis includens (Walker) (Lepidoptera: Noctuidae), Heliothis virescens (F.) (Lepidoptera: Noctuidae), and Spodoptera cosmioides (Walker) (Lepidoptera: Noctuidae) during vegetative (V4) and reproductive (R2 and R4) crop developmental stages. The efficacy of DAS-81419-2 was compared to that of a non-Bt isogenic variety managed with or without applications of commercial foliar insecticides for lepidopteran control. DAS-81419-2 soybean consistently experienced defoliation levels of 0.5% or less (compared with 20.05-56.74% in the non-Bt, nonsprayed treatment) and larval survival of < 0.1% in all four species across the vegetative and reproductive plant stages evaluated. The efficacy of DAS-81419-2 was significantly higher than commercial foliar insecticides applied to the non-Bt variety. DAS-81419-2 soybeans containing two highly effective Bt proteins are expected to be a more robust IRM tool compared to single-trait Bt technologies. The consistent efficacy of DAS-81419-2 soybeans across years, locations, and crop stages suggests that it will be a valuable product for management of hard-to-control key lepidopteran pests in South American soybean production.

Partial Life History of Chrysodeixis includens (Lepidoptera: Noctuidae) on Summer Hosts.

The soybean looper, Chrysodeixis includens (Walker) (Lepidoptera: Noctuidae), is a major defoliating pest of soybeans, Glycine max (L.) Merrill, in Louisiana. However, other alternate host crops in the agroecosystem have the potential to impact C. includens populations. Life table statistics of C. includens on four host plants were evaluated. C. includens larvae were fed leaves of three cotton Gossypium hirsutum L. cultivars 'DP 143 B2RF,' 'DP 174 RF,' and 'PHY 485 WRF'; cowpea Vigna unguiculata (L.) Walpers 'California Blackeye'; three soybean cultivars 'Lyon,' 'PI 227687,' and 'RC 4955'; and sweetpotato Ipomoea batatas (L.) Lamarck 'Evangeline.' All C. includens larvae reared on cotton cultivars DP 143 B2RF and PHY 485 WRF experienced 100% mortality during the first instar. Total developmental period of preadult C. includens was significantly shorter on cotton DP 174 RF and cowpea California Blackeye but longer on sweetpotato Evangeline. Sweetpotato Evangeline had the highest amount of leaf tissue consumed and soybean Lyon had the least. Pupal weight was highest when insects fed on cotton DP 174 RF and lowest on soybean PI 227687. Life table statistics showed that the highest intrinsic rate of increase and net reproductive rate were attained when insects were reared on cotton DP 174 RF and cowpea California Blackeye whilst the lowest were recorded on soybean PI 227687. This study provides valuable information on the role of alternative host crops on the partial life history of C. includens in Louisiana agroecosystems.