Resistance of Bt and Non-Bt Soybean Cultivars Adapted to Novel Growing Regions of Brazil to Chrysodeixis includens and Spodoptera frugiperda.

Soybean is a highly valuable commodity crop for Brazil's economy. However, it faces significant threats from the attack of a complex of lepidopteran pests, particularly Chrysodeixis includens (Walker) and Spodoptera frugiperda (J. E. Smith). These pests have been managed primarily using transgenic Bt soybeans, but limited knowledge exists about the resistance levels of Bt and non-Bt cultivars adapted to novel soybean-growing areas in Brazil, such as the Minas Gerais state. This study evaluated the resistance levels of Bt and non-Bt soybean cultivars to C. includens and S. frugiperda, and whether the Bt cultivars can differentially affect these pests across larval stages. No-choice bioassays were conducted using Bt (NS6010 IPRO and P97R50 IPRO) and non-Bt soybeans (UFLA 6301 RR, P96R90 RR, and ANsc 80111 RR) at V4-stage in the laboratory with neonate (24 h) and third-instar larvae. Larvae were fed leaf discs in Petri dishes, recording the mortality, leaf consumption, and weight gain after 7 days. There was high mortality of C. includens neonates on the Bt cultivars, but this trend was not observed for older larvae. For S. frugiperda neonates, there was high mortality on the Bt cultivar NS 6010 IPRO and non-Bt cultivar UFLA 6301 RR, but only the former was effective for older larvae. Although the Bt cultivars did not kill the third instars, antinutritional effects were found, such that leaf tissue consumed was not converted to larval weight gain. These findings are important for defining regional strategies of integrated and resistance management of C. includens and S. frugiperda in expanding regions of soybean cultivation in Brazil.

Characterizing the differential susceptibility and resistance to insecticides in populations of Chrysodeixis includens and Rachiplusia nu (Lepidoptera: Noctuidae) in Brazil.

BACKGROUND: Chrysodeixis includens (Walker) and Rachiplusia nu (Guenée) are major Plusiinae pests of soybean in the Southern Cone region of South America. In recent decades, C. includens was the main defoliator of soybean in Brazil, but from 2021 onwards, R. nu emerged as an important soybean pest in various regions of the country. Here, we characterize the differential susceptibility and resistance to insecticides in these Plusiinae pests from two soybean regions of Brazil. RESULTS: Except for spinetoram and chlorfenapyr (comparable lethality against both species) and a Bt-based biopesticide (more lethal for C. includens), the tested insecticides showed higher lethality against R. nu than against C. includens, but populations of the same species, even separated by long distances, presented similar resistance levels. For both species, the 90% lethal concentration (LC90) values of most insecticides were higher than the field-recommended dose. Nevertheless, the field-recommended doses of spinetoram, metaflumizone, emamectin benzoate, cyclaniliprole and chlorfenapyr showed comparable control efficacy against both species, whereas indoxacarb, chlorantraniliprole, flubendiamide, teflubenzuron and chlorfluazuron were more lethal for R. nu, and methoxyfenozide and the Bt-based insecticide were more lethal for C. includens. Thiodicarb, methomyl and lambda-cyhalothrin showed low lethality against both species. CONCLUSIONS: Large interspecific differences in the susceptibility to insecticides was found in major Plusiinae pests of soybean in Brazil. Furthermore, variations in susceptibility to insecticides occurred consistently among species and populations, regardless of the collection site and thus despite unequal temporal and spatial exposure to insecticides. These results demonstrate that accurate species identification is essential for effective control of Plusiinae in soybean. © 2024 Society of Chemical Industry.

Oviposition behavior and foliar consumption of Chrysodeixis includes (Lepidoptera: Noctuidae) in soybean genotypes / Comportamento de oviposição e consumo foliar de Chrysodeixis includens (Lepidoptera: Noctuidae) em genótipos de soja

The use of plant resistance acts by intervening in the herbivore-host relationship, through morphological, physical or chemical factors of the plant. This study evaluated the oviposition and foliar consumption of Chrysodeixis includens (Walker [1858]) in soybean genotypes, in free- and no-choice tests, correlating them with the factors, density and size of trichomes. The experiments were carried out in laboratory (25 ± 2 °C; RH= 70 ± 10%; photoperiod 14h) using five cultivars (BRS 391, BRS 6203 RR, BMX Valente RR, Tec Irga 6070 RR, BMX Icone Ipro) and two isolines (PELBR 10-6000 and PELBR 10-6049). The trichomes reported were filiform tectors and claviform multicellular glandular. The density of glandular trichomes, in stages V2 and V5, was higher on BRS 6203 RR and BRS 391, respectively. The higher density of glandular trichomes was observed in V5 and a higher density of tector trichomes in V2. The lowest densities and the smallest sizes of trichomes in V2 and V5 stages was observed on PELBR 10-6049. The size of tector trichomes and the number of eggs did not differ among the cultivars. Foliar consumption was lower for on BMX Icone Ipro and Tec Irga 6070 RR. Trichome density influences the consumption and oviposition behavior of C. includens.

O uso da resistência de plantas atua intervindo na relação herbívoro-hospedeiro, através de fatores morfológicos, físicos ou químicos da planta. O objetivo deste trabalho foi avaliar a oviposição e o consumo foliar de Chrysodeixis includens (Walker [1858]) em genótipos de soja, através de testes com e sem escolha, correlacionando-os com os fatores, densidade e tamanho de tricomas. Os experimentos foram realizados em laboratório (25 ± 2 °C; UR= 70 ± 10%; fotoperíodo 14h) utilizando cinco cultivares (BRS 391, BRS 6203 RR, BMX Valente RR, Tec Irga 6070 RR, BMX Icone Ipro) e duas linhagens (PELBR 10-6000 e PELBR 10-6049). Os tricomas encontrados foram tectores filiformes e glandulares multicelulares claviformes. A densidade de tricomas glandulares nos estágios V2 e V5, foi maior em BRS 6203 RR e BRS 391, respectivamente. A maior densidade de tricomas glandulares foi observada em V5 e a maior densidade de tricomas tectores em V2. As menores densidades e os menores tamanhos de tricomas nos estágios V2 e V5 foram observados em PELBR 10-6049. O tamanho de tricomas tectores e o número de ovos não diferiram entre as cultivares. O consumo foliar foi menor para BMX Icone Ipro e Tec Irga 6070 RR. A densidade de tricomas influência o comportamento de consumo e oviposição de C. includens.

Development and biological evaluation of nanoencapsulated-based pyrethroids with synergists for resistance management of two soybean pests: insights for new insecticide formulations.

BACKGROUND: Chemical control is commonly used against Euschistus heros (F.) and Chrysodeixis includens (Walker) in soybean fields in South America. However, previous studies reported that these pests have reduced susceptibility to pyrethroids in Brazil. On this basis, we developed and evaluated nanoencapsulated-based bifenthrin (BFT) and λ-cyhalothrin (LAM) with the synergists piperonyl butoxide (PBO) and diethyl maleate (DEM) for insect resistance management (IRM). RESULTS: Nanoformulations of BFT and LAM with PBO and DEM presented good physical-chemical characteristics and were stable. The spherical morphology of all systems and the encapsulation efficiency in nanostructured lipid carriers did not change when synergists were added. Nanoencapsulated BFT with DEM applied topically increased the susceptibility of E. heros to BFT by 3.50-fold. Similarly, nanoencapsulated BFT and LAM with PBO in diet-overlay bioassays increased the susceptibility of C. includens to both chemicals by up to 2.16-fold. Nanoencapsulated BFT and LAM with synergists also improve control efficacy of both species, causing higher mortality than commercial products containing these chemistries. CONCLUSIONS: It is possible to develop nanoencapsulated-based formulations of BFT and LAM with PBO or DEM, and these nanoformulations have the potential to improve control of E. heros and C. includens with recognized low susceptibility to pyrethroids. This study provides updates for designing new insecticide formulations for IRM. © 2022 Society of Chemical Industry.

Bioactivity of essential oils from Croton grewioides and its major compounds: toxicity to soybean looper Chrysodeixis includens and selectivity to the predatory stink bug Podisus nigrispinus.

Natural biological control is a key factor that ensures the regulation of insect pest populations in agroecosystems. However, the indiscriminate use of pesticides has compromised this environmental service. Thus, the search for environmentally safe pesticides is an increasing requirement for sustainable food production. In this study, we analyzed the toxicity of essential oils from two accessions (CGR112 and CGR126) of Croton grewioides and its major compounds, methyl eugenol and eugenol, on the soybean pest Chrysodeixis includens. In addition, we investigated the sublethal effects of these compounds on the predatory bug Podisus nigrispinus, analyzing its developmental, reproduction and life table parameters. Essential oils and their major compounds were toxic to C. includens and P. nigrispinus. In general, the presence of eugenol made the essential oil more toxic to the pest and selective to the natural enemy. Eugenol was the most toxic compound for 2nd instar larvae of C. includens at LD50, followed by CGR126 essential oil from C. grewioides which was equally toxic at higher doses. The estimated lethal times for essential oils to cause mortality in 50% of the population of C. includens were less than 15 h. There was selectivity of the essential oil of CGR126 accession of C. grewioides at lethal doses above 90%. Although the treatments showed little effect on the development of P. nigrispinus, body mass and reproductive parameters were negatively affected, with the exception of the essential oil of CGR126 accession of C. grewioides. The essential oil of C. grewioides may be a promising active ingredient for the synthesis of new insecticides, which are efficient against C. includens and at the same time are safer for the natural enemy P. nigrispinus.

The role of glyphosate-resistant weeds and starvation on biological, reproductive, and preference parameters of Chrysodeixis includens (Lepidoptera: Noctuidae).

Glyphosate-resistant weeds are difficult to manage and can serve as hosts for pests that threaten cultivated crops. Chrysodeixis includens (Walker) (Lepidoptera: Noctuidae) is one of the main polyphagous pests of soybean in Brazil that can benefit from weeds' presence during season and off-season. Despite its pest status, little is known about C. includens survival and development on alternative hosts, including those resistant to glyphosate. Therefore, we assessed the biology, reproduction, preference, and survival at different feeding periods of C. includens on seven glyphosate-resistant weeds (Sumatran fleabane, Italian ryegrass, sourgrass, goosegrass, smooth pigweed, wild poinsettia, hairy beggarticks) commonly found in Brazilian agroecosystems, under laboratory conditions. Our results showed that C. includens survival and reproduction were similar on soybean and wild poinsettia. Survival and reproduction were lower on smooth pigweed and hairy beggarticks. Also, these plants prolonged the larval stage. Larvae did not pupate when fed on sourgrass, goosegrass, Italian ryegrass, and Sumatran fleabane. However, on Sumatran fleabane their biomass was higher. The mean generation time was lower on wild poinsettia. This weed was preferred to soybean. An antifeeding factor was observed on Sumatran fleabane. Larvae fed for 11 days on soybean, wild poinsettia and smooth pigweed developed into pupae. In agricultural systems, farmers must pay attention to the management of these weeds, especially wild poinsettia, smooth pigweed, and hairy beggarticks, to interrupt the cycle of this pest, since these plants can serve as main sources of infestation for the soybean crop.

The Effect of Synergistic Compounds on the Susceptibility of Euschistus heros (Hemiptera: Pentatomidae) and Chrysodeixis includens (Lepidoptera: Noctuidae) to Pyrethroids.

The Neotropical brown stink bug, Euschistus heros (F.), and the soybean looper, Chrysodeixis includens (Walker), are key pests of soybean in South America. Low susceptibility to pyrethroids has been reported for both species in Brazil. Here, we evaluate the addition of synergistic compounds piperonyl butoxide (PBO) and diethyl maleate (DEM) to manage E. heros and C. includens with resistance to λ-cyhalothrin and bifenthrin. The LD50 of technical grade and commercial products containing λ-cyhalothrin and bifenthrin decreased against field-collected E. heros exposed to PBO and DEM relative to unexposed insects; synergistic ratios up to 4.75-fold. The mortality also increased when E. heros were exposed to commercial formulations containing λ-cyhalothrin (from 4 to 44%) and bifenthrin (from 44 to 88%) in the presence of synergists. There was also a higher susceptibility of field-collected C. includens to technical grade λ-cyhalothrin when PBO was used; synergistic ratio of 5.50-fold. High lethally of technical grade λ-cyhalothrin was also verified in the presence of PBO, with mortality increasing from 6 to 57%. Our findings indicate the potential utility of synergists in reversing the resistance to λ-cyhalothrin and bifenthrin in E. heros and C. includens and suggest a significant role of metabolic mechanisms underlying the detoxification of both pyrethroids.

Interspecific Variation in Susceptibility to Insecticides by Lepidopteran Pests of Soybean, Cotton, and Maize Crops From Brazil.

The interspecific variation in susceptibility to insecticides by lepidopteran species of soybean [Glycine max L. (Merr.)], cotton (Gossypium hirsutum L.), and maize (Zea mays L.) crops from Brazil were evaluated. Populations of Anticarsia gemmatalis (Hübner) (Lepidoptera: Erebidae), Chrysodeixis includens (Walker), Helicoverpa armigera (Hübner), Spodoptera frugiperda (Smith), Spodoptera eridania (Stoll), Spodoptera cosmioides (Walker), and Spodoptera albula (Walker) (Lepidoptera: Noctuidae) were collected from 2019 to 2021. Early L3 larvae (F2 generation) were exposed to the formulated insecticides methoxyfenozide, indoxacarb, spinetoram, flubendiamide, and chlorfenapyr in diet-overlay bioassays. The median lethal concentrations (LC50) were used to calculate tolerance ratios (TR) of each species in relation to the most susceptible species to each insecticide. The lowest LC50 values were verified for A. gemmatalis to all insecticides tested. Chrysodeixis includens and most of the Spodoptera species were moderately tolerant to methoxyfenozide (TR < 8.0-fold) and indoxacarb (TR < 39.4-fold), whereas H. armigera was the most tolerant species to methoxyfenozide (TR = 21.5-fold), and indoxacarb (TR = 106.4-fold). Spodoptera cosmioides, S. eridania, and S. albula showed highest tolerance to spinetoram (TR > 1270-fold), S. eridania, S. frugiperda, and S. albula to flubendiamide (TR from 38- to 547-fold), and S. albula to indoxacarb (TR = 138.6-fold). A small variation in susceptibility to chlorfenapyr (TR < 4.4-fold) was found among the lepidopteran evaluated. Our findings indicate a large variation in susceptibility to indoxacarb, spinetoram, and flubendiamide and a relatively low variation in susceptibility to methoxyfenozide and chlorfenapyr by lepidopteran species of soybean, cotton, and maize from Brazil.

Susceptibility of Alabama argillacea and Chrysodeixis includens (Lepidoptera: Noctuidae) larvae to Beauveria bassiana associated with kaolin / Susceptibilidade de larvas de Alabama argillacea e Chrysodeixis includens (Lepidoptera: Noctuidae) a Beauveria bassiana associada com caulim

The mortality of the Alabama argillacea and Chrysodeixis includens (Lepidoptera: Noctuidae) larvae caused by the kaolin inert powder and the entomopathogenic fungus Beauveria bassiana were determined under laboratory conditions. Using the caterpillar submersion method, the CG 138 B. bassiana isolate was more pathogenic to A. argillacea than the CG 70, GC 82, ESALQ 634, and ESALQ 645. All five tested isolates caused similar mortality of C. includens. The mortality of first-instar larvae of A. argillacea and C. includens by feeding on leaf-disc impregnated with B. bassiana (CG 138) and kaolin was also determined. Higher A. argillacea mortalities were observed in the B. bassiana (CG 138) treatments, regardless of the presence of kaolin. However, the activity of kaolin + B. bassiana (CG 138) against C. includens was higher than each ingredient alone, indicating an additive action against C. includes larvae. The mortality of A. argillacea and C. includens larvae treated with kaolin + B. bassiana (CG 138) was similar, and the A. argillacea mortality was higher than that of C. includens with kaolin and B. bassiana (GC 138) separated. The treatment kaolin + B. bassiana (CG 138) is promising for the simultaneous management of these two defoliator pests, mainly A. includes. In addition, the monophagous A. argillacea is more susceptible to both kaolin and B. bassiana (GC 138) than the polyphagous C. includens, suggesting that the nutritional ecology plays an important role in the susceptibility of these defoliator species to alternative insecticides.(AU)

A mortalidade de larvas de Alabama argillacea e Chrysodeixis includens (Lepidoptera: Noctuidae), causada pelo pó inerte do caulim e pelo fungo entomopatogênico Beauveria bassiana, foi determinada em condições de laboratório. Usando o método de submersão da lagarta, o isolado CG 138 de B. bassiana foi mais patogênico para A. argillacea que os CG 70, GC 82, ESALQ 634 e ESALQ 645 desse fungo. Os cinco isolados testados causaram mortalidade semelhante de C. includens. A mortalidade de larvas de primeiro ínstar de A. argillacea e C. includens pelo método de alimentação em discos foliares impregnados com B. bassiana (CG 138) e caulim foi, também, determinada. A mortalidade de A. argillacea foi maior nos tratamentos com B. bassiana (CG 138), independentemente da presença do caulim. A atividade do caulim + B. bassiana (CG 138) contra C. includens foi maior que cada ingrediente isolado, indicando ação aditiva contra larvas desse Lepidoptera. A mortalidade de larvas de A. argillacea e C. includens, com caulim + B. bassiana (CG 138), foi semelhante e as de A. argillacea maiores que as de C. includens com caulim e B. bassiana (GC 138) isoladamente. O tratamento caulin + B. bassiana (CG 138) é promissor para o manejo simultâneo dessas duas pragas desfolhadoras, principalmente A. includens. Além disso, a monófaga, A. argilacea, é mais suscetível ao caulim e à B. bassiana (GC 138) que a polífaga, C. includens, sugerindo que a ecologia nutricional desempenha papel importante na suscetibilidade dessas espécies desfolhadoras a inseticidas alternativos.(AU)

Susceptibility of Alabama argillacea and Chrysodeixis includens (Lepidoptera: Noctuidae) larvae to Beauveria bassiana associated with kaolin / Susceptibilidade de larvas de Alabama argillacea e Chrysodeixis includens (Lepidoptera: Noctuidae) a Beauveria bassiana associada com caulim

Abstract The mortality of the Alabama argillacea and Chrysodeixis includens (Lepidoptera: Noctuidae) larvae caused by the kaolin inert powder and the entomopathogenic fungus Beauveria bassiana were determined under laboratory conditions. Using the caterpillar submersion method, the CG 138 B. bassiana isolate was more pathogenic to A. argillacea than the CG 70, GC 82, ESALQ 634, and ESALQ 645. All five tested isolates caused similar mortality of C. includens. The mortality of first-instar larvae of A. argillacea and C. includens by feeding on leaf-disc impregnated with B. bassiana (CG 138) and kaolin was also determined. Higher A. argillacea mortalities were observed in the B. bassiana (CG 138) treatments, regardless of the presence of kaolin. However, the activity of kaolin + B. bassiana (CG 138) against C. includens was higher than each ingredient alone, indicating an additive action against C. includes larvae. The mortality of A. argillacea and C. includens larvae treated with kaolin + B. bassiana (CG 138) was similar, and the A. argillacea mortality was higher than that of C. includens with kaolin and B. bassiana (GC 138) separated. The treatment kaolin + B. bassiana (CG 138) is promising for the simultaneous management of these two defoliator pests, mainly A. includes. In addition, the monophagous A. argillacea is more susceptible to both kaolin and B. bassiana (GC 138) than the polyphagous C. includens, suggesting that the nutritional ecology plays an important role in the susceptibility of these defoliator species to alternative insecticides.

Resumo A mortalidade de larvas de Alabama argillacea e Chrysodeixis includens (Lepidoptera: Noctuidae), causada pelo pó inerte do caulim e pelo fungo entomopatogênico Beauveria bassiana, foi determinada em condições de laboratório. Usando o método de submersão da lagarta, o isolado CG 138 de B. bassiana foi mais patogênico para A. argillacea que os CG 70, GC 82, ESALQ 634 e ESALQ 645 desse fungo. Os cinco isolados testados causaram mortalidade semelhante de C. includens. A mortalidade de larvas de primeiro ínstar de A. argillacea e C. includens pelo método de alimentação em discos foliares impregnados com B. bassiana (CG 138) e caulim foi, também, determinada. A mortalidade de A. argillacea foi maior nos tratamentos com B. bassiana (CG 138), independentemente da presença do caulim. A atividade do caulim + B. bassiana (CG 138) contra C. includens foi maior que cada ingrediente isolado, indicando ação aditiva contra larvas desse Lepidoptera. A mortalidade de larvas de A. argillacea e C. includens, com caulim + B. bassiana (CG 138), foi semelhante e as de A. argillacea maiores que as de C. includens com caulim e B. bassiana (GC 138) isoladamente. O tratamento caulin + B. bassiana (CG 138) é promissor para o manejo simultâneo dessas duas pragas desfolhadoras, principalmente A. includens. Além disso, a monófaga, A. argilacea, é mais suscetível ao caulim e à B. bassiana (GC 138) que a polífaga, C. includens, sugerindo que a ecologia nutricional desempenha papel importante na suscetibilidade dessas espécies desfolhadoras a inseticidas alternativos.

Mixture compatibility of ChinNPV baculovirus with herbicides and fungicides used in soybean / Compatibilidade de mistura do baculovírus ChinNPV com herbicidas e fungicidas utilizados na soja

Chrysodeixis includens Walker, 1858 (Lepidoptera: Noctuidae), commonly known as soybean looper, is one of the main pests that reduces soybean yield due to its defoliation capacity. Population outbreaks of this pest can occur in the vegetative stage of the crop, together with the occurrence of weeds or in the reproductive stage, at the same time as that of diseases. This often requires the use of pesticides against pests and weeds, or fungi at the same time. Thus, the objective of this study was to evaluate the compatibility of baculovirus ChinNPV with different synthetic chemical herbicides and fungicides used in soybean. Four bioassays were carried out, with or without the addition of ChinNPV to different herbicides and fungicides. The artificial diets were immersed in the solutions of the pesticides and their mixture and supplied to the caterpillars of C. includens, immediately and after one and two hours of mixing. The evaluation was performed by quantifying the number of dead caterpillars. The results showed that the ChinNPV baculovirus is compatible for mixing with all studied herbicides and fungicides, even after two hours of mixing. In all the studied scenarios, baculovirus mixed with chemical pesticides triggered the same mortality as sprayed with baculovirus alone, and values of over 80% mortality of C. includens have always been recorded.

Chrysodeixis includens Walker, 1858 (Lepidoptera: Noctuidae) falsa-medideira é uma das principais pragas que reduz o rendimento da soja devido à sua capacidade de desfolha. Surtos populacionais desta praga podem ocorrer no estágio vegetativo da cultura, juntamente com a ocorrência de ervas daninhas ou no estágio reprodutivo, ao mesmo tempo em que ocorrem doenças. Muitas vezes, isso pode exigir o uso de pesticidas contra pragas e ervas daninhas ou fungos ao mesmo tempo. Assim, o objetivo deste trabalho foi avaliar a compatibilidade do baculovírus ChinNPV com diferentes herbicidas e fungicidas químicos sintéticos utilizados na soja. Foram realizados quatro bioensaios, com ou sem adição de ChinNPV em diferentes herbicidas e fungicidas. As dietas artificiais foram imersas nas soluções dos agrotóxicos e sua mistura e fornecidas às lagartas de C. includens, imediatamente e após uma e duas horas de mistura. A avaliação foi realizada quantificando o número de lagartas mortas. Os resultados mostraram que o baculovírus ChinNPV é compatível para mistura com todos os herbicidas e fungicidas estudados mesmo após duas horas de mistura. Em todos os cenários estudados, o baculovírus misturado com pesticidas químicos desencadeou a mesma mortalidade que o baculovírus pulverizado sozinho, e valores de mais de 80% da mortalidade de C. includens sempre foram registrados.

Mixture compatibility of ChinNPV baculovirus with herbicides and fungicides used in soybean / Compatibilidade de mistura do baculovírus ChinNPV com herbicidas e fungicidas utilizados na soja

Chrysodeixis includens Walker, 1858 (Lepidoptera: Noctuidae), commonly known as soybean looper, is one of the main pests that reduces soybean yield due to its defoliation capacity. Population outbreaks of this pest can occur in the vegetative stage of the crop, together with the occurrence of weeds or in the reproductive stage, at the same time as that of diseases. This often requires the use of pesticides against pests and weeds, or fungi at the same time. Thus, the objective of this study was to evaluate the compatibility of baculovirus ChinNPV with different synthetic chemical herbicides and fungicides used in soybean. Four bioassays were carried out, with or without the addition of ChinNPV to different herbicides and fungicides. The artificial diets were immersed in the solutions of the pesticides and their mixture and supplied to the caterpillars of C. includens, immediately and after one and two hours of mixing. The evaluation was performed by quantifying the number of dead caterpillars. The results showed that the ChinNPV baculovirus is compatible for mixing with all studied herbicides and fungicides, even after two hours of mixing. In all the studied scenarios, baculovirus mixed with chemical pesticides triggered the same mortality as sprayed with baculovirus alone, and values of over 80% mortality of C. includens have always been recorded.(AU)

Chrysodeixis includens Walker, 1858 (Lepidoptera: Noctuidae) falsa-medideira é uma das principais pragas que reduz o rendimento da soja devido à sua capacidade de desfolha. Surtos populacionais desta praga podem ocorrer no estágio vegetativo da cultura, juntamente com a ocorrência de ervas daninhas ou no estágio reprodutivo, ao mesmo tempo em que ocorrem doenças. Muitas vezes, isso pode exigir o uso de pesticidas contra pragas e ervas daninhas ou fungos ao mesmo tempo. Assim, o objetivo deste trabalho foi avaliar a compatibilidade do baculovírus ChinNPV com diferentes herbicidas e fungicidas químicos sintéticos utilizados na soja. Foram realizados quatro bioensaios, com ou sem adição de ChinNPV em diferentes herbicidas e fungicidas. As dietas artificiais foram imersas nas soluções dos agrotóxicos e sua mistura e fornecidas às lagartas de C. includens, imediatamente e após uma e duas horas de mistura. A avaliação foi realizada quantificando o número de lagartas mortas. Os resultados mostraram que o baculovírus ChinNPV é compatível para mistura com todos os herbicidas e fungicidas estudados mesmo após duas horas de mistura. Em todos os cenários estudados, o baculovírus misturado com pesticidas químicos desencadeou a mesma mortalidade que o baculovírus pulverizado sozinho, e valores de mais de 80% da mortalidade de C. includens sempre foram registrados.(AU)

Optimization of Large-Scale Production of Chrysodeixis includens nucleopolyhedrovirus for Its Use as a Biopesticide.

The baculovirus Chrysodeixis includens nucleopolyhedrovirus (ChinNPV) is pathogenic to Chrysodeixis includens (Walker) (Lepidoptera: Noctuidae) larvae, known as soybean looper, which is an important pest of soybean and bean. In this study, some parameters were tested to overcome the difficulties in the in vivo production of ChinNPV aiming to increase its use as a biopesticide. First, different combinations of larval instars (3rd and 4th instars), larval incubation temperatures (23 °C and 26 °C), and rearing densities (individually and 10 larvae/cup) were compared for larval weight and the production of occlusion bodies (OBs). A positive correlation (p< 0.001) was observed for OB production and larval weight. Fourth instar larvae produced more OBs than third instar larvae (p<0.05); however, no significant differences in OBs/larva (p>0.05) were observed for larvae kept in groups or individually. Therefore, a second assay was performed using fourth instar larvae incubated at 26 °C and two larval densities (10 larvae/cup and 40 larvae/cup). The losses of insects and OB production were evaluated as well as the influence of storage temperatures post-mortem (-20 °C, 4 °C, and 15 °C) in the OB yield. As expected, insect losses due to cannibalism or microbial contamination were greater (p<0.05) with the increase in larval density, although no difference was observed in OBs/larva (p>0.05). In addition, the storage temperature post-mortem did not influence the OB yield (p>0.05). The average production of ChinNPV OBs was 3×1010 OBs/40 larvae cup. The results demonstrate the viability of rearing C. includens in groups to enhance the mass production and reduce virus production costs.

Distribution and invasion risk assessments of Chrysodeixis includens (Walker, [1858]) (Lepidoptera: Noctuidae) using CLIMEX.

Chrysodeixis includens is a polyphagous pest restricted to the American continent. The occurrence of C. includens is allied, among other factors, by favorable conditions such as temperature, humidity, presence of hosts, and migratory behavior. In this work, we built spatiotemporal species distribution models at continental and global levels for the distribution of C. includens using CLIMEX to determine times and regions favorable for year-round survival and migration of this species and in case of invasion on other continents to apply timely and right phytosanitary measures. Our models estimated high climate suitability for C. includens in Central and large proportions of South America throughout the year. Moreover, there is suitability for C. includens growth in all months of the year in Central and northern part of South America. In the northern hemisphere, these conditions range from April to October, while in mid-southern parts of South America, favorable periods comprise October through June. The countries with the highest suitability for C. includens outside the American continent are located on the African and Asian continents. Our results show variable climate suitability for C. includens during the year that help to understand likely migration pattern in North America. This information would direct efforts for appropriate C. includens management during warm and moist periods of the year. Furthermore, our models notify the need for the development of strategies for the inspection and interception of C. includens especially in central Africa, India, South and Southeast Asia, and Northeast Australia.

Transcriptome Analysis of Pyrethroid-Resistant Chrysodeixis includens (Lepidoptera: Noctuidae) Reveals Overexpression of Metabolic Detoxification Genes.

Chrysodeixis includens (Walker, [1858]) is one of the most important defoliator of soybean in Brazil because of its extensive geographical distribution and high tolerance to insecticides compared with other species of caterpillars. Because of this, we conducted bioassays to evaluate the efficacy of pyrethroid λ-cyhalothrin on a C. includens resistant strain (MS) and a susceptible (LAB) laboratory strain. High throughput RNA sequencing (RNA-seq) of larval head and body tissues were performed to identify potential molecular mechanisms underlying pyrethroid resistance. Insecticide bioassays showed that MS larvae exhibit 28.9-fold resistance to pyrethroid λ-cyhalothrin relative to LAB larvae. RNA-seq identified evidence of metabolic resistance in the head and body tissues: 15 cytochrome P450 transcripts of Cyp6, Cyp9, Cyp4, Cyp304, Cyp307, Cyp337, Cyp321 families, 7 glutathione-S-transferase (Gst) genes, 7 α-esterase genes from intracellular and secreted catalytic classes, and 8 UDP-glucuronosyltransferase (Ugt) were overexpressed in MS as compared with LAB larvae. We also identified overexpression of GPCR genes (CiGPCR64-like and CiGPCRMth2) in the head tissue. To validate RNA-seq results, we performed RT-qPCR to assay selected metabolic genes and confirmed their expression profiles. Specifically, CiCYP9a101v1, CiCYP6ae149, CiCYP6ae106v2, CiGSTe13, CiCOE47, and CiUGT33F21 exhibited significant overexpression in resistant MS larvae. In summary, our findings detailed potential mechanisms of metabolic detoxification underlying pyrethroid resistance in C. includens.

Performance of Genetically Modified Soybean Expressing the Cry1A.105, Cry2Ab2, and Cry1Ac Proteins Against Key Lepidopteran Pests in Brazil.

The pyramided genetically modified (GM) soybean [Glycine max L. (Merr.)] MON87751 × MON87708 × MON87701 × MON89788, expressing Cry1A.105, Cry2Ab2, and Cry1Ac from Bacillus thuringiensis Berliner, was approved for commercial use in Brazil. We conducted laboratory, greenhouse, and field studies to assess the efficacy of this Bt soybean against key soybean lepidopteran pests. Neonates of Anticarsia gemmatalis (Hübner) (Lepidoptera: Erebidae), Chrysodeixis includens (Walker), and Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) were exposed to Bt proteins in diet-overlay bioassays. MON87751 × MON87708 × MON87701 × MON89788 soybean and individual components were evaluated in laboratory (leaf disc), greenhouse (high artificial infestations), and in field conditions (natural infestations). Neonates of A. gemmatalis, C. includens, and H. armigera were highly susceptible to Cry1A.105 (LC50 from 0.79 to 48.22 ng/cm2), Cry2Ab2 (LC50 from 1.24 to 8.36 ng/cm2), and Cry1Ac (LC50 from 0.15 to 5.07 ng/cm2) in diet-overlay bioassays. In laboratory leaf disc bioassays and greenhouse trials, MON87751 × MON87708 × MON87701 × MON89788 soybean as well as the individual components were highly effective in controlling A. gemmatalis, C. includens, and H. armigera. Similarly, under field conditions, the pyramided genotypes expressing Cry1A.105, Cry2Ab2, and Cry1Ac were highly effective at protecting soybean against C. includens. We concluded that the individual Bt proteins expressed by GM soybean MON87751 × MON87708 × MON87701 × MON89788 killed all or nearly all the susceptible A. gemmatalis, C. includens, and H. armigera, fulfilling one important criterion for successfully delaying resistance to pyramided Bt crops.

Susceptibility of Alabama argillacea and Chrysodeixis includens (Lepidoptera: Noctuidae) larvae to Beauveria bassiana associated with kaolin

Abstract The mortality of the Alabama argillacea and Chrysodeixis includens (Lepidoptera: Noctuidae) larvae caused by the kaolin inert powder and the entomopathogenic fungus Beauveria bassiana were determined under laboratory conditions. Using the caterpillar submersion method, the CG 138 B. bassiana isolate was more pathogenic to A. argillacea than the CG 70, GC 82, ESALQ 634, and ESALQ 645. All five tested isolates caused similar mortality of C. includens. The mortality of first-instar larvae of A. argillacea and C. includens by feeding on leaf-disc impregnated with B. bassiana (CG 138) and kaolin was also determined. Higher A. argillacea mortalities were observed in the B. bassiana (CG 138) treatments, regardless of the presence of kaolin. However, the activity of kaolin + B. bassiana (CG 138) against C. includens was higher than each ingredient alone, indicating an additive action against C. includes larvae. The mortality of A. argillacea and C. includens larvae treated with kaolin + B. bassiana (CG 138) was similar, and the A. argillacea mortality was higher than that of C. includens with kaolin and B. bassiana (GC 138) separated. The treatment kaolin + B. bassiana (CG 138) is promising for the simultaneous management of these two defoliator pests, mainly A. includes. In addition, the monophagous A. argillacea is more susceptible to both kaolin and B. bassiana (GC 138) than the polyphagous C. includens, suggesting that the nutritional ecology plays an important role in the susceptibility of these defoliator species to alternative insecticides.

Resumo A mortalidade de larvas de Alabama argillacea e Chrysodeixis includens (Lepidoptera: Noctuidae), causada pelo pó inerte do caulim e pelo fungo entomopatogênico Beauveria bassiana, foi determinada em condições de laboratório. Usando o método de submersão da lagarta, o isolado CG 138 de B. bassiana foi mais patogênico para A. argillacea que os CG 70, GC 82, ESALQ 634 e ESALQ 645 desse fungo. Os cinco isolados testados causaram mortalidade semelhante de C. includens. A mortalidade de larvas de primeiro ínstar de A. argillacea e C. includens pelo método de alimentação em discos foliares impregnados com B. bassiana (CG 138) e caulim foi, também, determinada. A mortalidade de A. argillacea foi maior nos tratamentos com B. bassiana (CG 138), independentemente da presença do caulim. A atividade do caulim + B. bassiana (CG 138) contra C. includens foi maior que cada ingrediente isolado, indicando ação aditiva contra larvas desse Lepidoptera. A mortalidade de larvas de A. argillacea e C. includens, com caulim + B. bassiana (CG 138), foi semelhante e as de A. argillacea maiores que as de C. includens com caulim e B. bassiana (GC 138) isoladamente. O tratamento caulin + B. bassiana (CG 138) é promissor para o manejo simultâneo dessas duas pragas desfolhadoras, principalmente A. includens. Além disso, a monófaga, A. argilacea, é mais suscetível ao caulim e à B. bassiana (GC 138) que a polífaga, C. includens, sugerindo que a ecologia nutricional desempenha papel importante na suscetibilidade dessas espécies desfolhadoras a inseticidas alternativos.

No cross-resistance between ChinNPV and chemical insecticides in Chrysodeixis includens (Lepidoptera: Noctuidae).

Chrysodeixis includens nucleopolyhedrovirus (ChinNPV: Baculoviridae: Alphabaculovirus) is an active ingredient of a biological-based insecticide (Chrysogen®) recommended against soybean looper (SBL), Chrysodeixis includens (Walker, [1858]), in soybean in Brazil. We investigated if SBL strains resistant to chemical insecticides are cross-resistant to the baculovirus ChinNPV. In droplet feeding bioassays, SBL strains resistant to lambda-cyhalothrin and teflubenzuron showed equivalent susceptibility to ChinNPV as heterozygous and susceptible strains, indicating no cross-resistance between ChinNPV and chemical insecticides in SBL. Therefore, the ChinNPV is a valuable new "mode-of-action" tool for SBL resistance management in Brazil.

Susceptibility of Brazilian Populations of Chrysodeixis includens (Lepidoptera: Noctuidae) to Selected Insecticides.

Soybean looper (SBL), Chrysodeixis includens (Walker) (Lepidoptera: Noctuidae), is an important pest of soybean and cotton in Brazil. The use of insecticides is one of the main control tactics against this pest. To support Integrated Pest Management (IPM) and Insect Resistance Management (IRM) programs, we characterized the susceptibility of Brazilian populations of SBL to insecticides. Field populations were collected from soybean fields during the 2016-2017 and 2017-2018 crop seasons. In the laboratory, late L2/early L3 larvae were exposed to insecticides in diet-overlay or topical bioassays. Field populations of SBL showed high susceptibility to spinetoram (LC50 = 0.074-0.25 µg a.i. per cm2), indoxacarb (LC50 = 0.46-0.94 µg a.i. per cm2), thiodicarb (LC50 = 9.14-36.61 µg a.i. per cm2), chlorantraniliprole (LC50 = 0.15-0.57 µg a.i. per cm2), flubendiamide (LC50 = 0.45-2.01 µg a.i. per cm2), and chlorfenapyr (LC50 = 0.15-0.25 µg a.i. per cm2); the resistance ratios were less than 16-fold. In contrast, SBL populations have reduced susceptibility to lambda-cyhalothrin (LC50 = 3.71-9.54 µg a.i./cm2), methoxyfenozide (LC50 = 0.67-4.23 µg a.i. per cm2), novaluron (LC50 = 27.52-77.63 µg a.i. per cm2), and teflubenzuron (LC50 = 13.41-73.02 µg a.i. per cm2). The resistance ratios relative to a Lab population (susceptible of reference) was up to 38-, 63-, 1,553-, and 5,215-fold, respectively. These high resistance ratios can be associated with resistance evolution. Implications of these findings to IPM and IRM programs are discussed.

Control of Chrysodeixis includens (Lepidoptera: Noctuidae) using Chin-IA (I-A) isolate as integrate component of management in soybean crops / Controle de Chrysodeixis includens (Lepidoptera: Noctuidae) usando o isolado Chin-IA (I-A) como componente do manejo integrado da cultura da soja

Chrysodeixis includens is an important pest of soybean crop who has gained more visibility in the Brazilian Cerrado due to damage caused in this region. Foliar consumption, feeding period and mortality level of soybean loopers in laboratory, as well as their control in the field conditions, were evaluated after application of the ChinNPV virus in soybean plants. In the laboratory, were tested six concentrations of isolate Chin-IA (I-A) (1 × 1011, 2 × 1011, 4 × 1011, 6 × 1011, 8 × 1011 and 10 × 1011 PIB ha-1), one dose of methomyl chemical insecticide (172 g ai ha-1) and distilled water (control). The field experiment was carried out in the 2016/2017 season using the same cultivar and laboratory treatments, except for the lowest virus concentration. The population density of small and large larvae was evaluated before and at 5, 8 and 12 days after application (DAA) of the treatments in soybean plants. All concentrations of the isolate Chin-IA (I-A) have reduced the soybean loopers consumption and their feeding period, showing 100% of mortality after 3 ­ 4 days without differing from treatment with the chemical insecticide. After eight DAA of virus in the field, the population density of small and large larvae was reduced, providing satisfactory levels of control. These results showed the evident potential of ChinNPV in the reduction of defoliation power and maintenance the soybean loopers population under of control level, and thus may be used as complementary method in the integrated management of this pest in soybean crops.(AU)

Chrysodeixis includens é uma importante praga da cultura da soja que tem ganhado maior visibilidade na região do Cerrado brasileiro em razão do dano causado. Consumo foliar, período de alimentação e mortalidade de lagartas falsa-medideira foram avaliados em laboratório, assim como seu controle em condições de campo, após a aplicação do vírus ChinNPV em plantas de soja. Em laboratório, foram testadas seis concentrações do isolado Chin-IA (I-A) (1 × 1011, 2 × 1011, 4 × 1011, 6 × 1011, 8 × 1011 e 10 × 1011 CIP ha-1), uma dose do inseticida químico metomil (172 g i.a ha-1) e água destilada (controle). O ensaio de campo foi realizado na safra 2016/2017 utilizando o mesmo cultivar e tratamentos do ensaio em laboratório, exceto para a menor concentração do vírus. A densidade populacional de lagartas pequenas e grandes foi avaliada antes (pré-contagem) e 5, 8 e 12 dias após a aplicação dos tratamentos nas plantas de soja. Todas as concentrações do isolado Chin-IA (I-A) reduziram o consumo e o período de alimentação das lagartas falsa-medideira, mostrando 100% de mortalidade entre 3 ­ 4 dias, sem diferir do tratamento com o inseticida químico. Depois de oito dias após a aplicação do vírus no campo, a densidade populacional de lagartas pequenas e grandes foi reduzida, promovendo um controle satisfatório. Esses resultados mostram o evidente potencial de ChinNPV na redução do poder de desfolha, mantendo a população das lagartas falsa-medideira abaixo do nível de controle, e indicam o seu uso como um método complementar no manejo integrado dessa praga em culturas de soja.(AU)