Anticarsia Gemmatalis Research Articles

Comparative effectiveness of Metarhizium rileyi, novaluron, and glyphosate on immune system, development, and redox metabolism of Anticarsia gemmatalis

Anticarsia gemmatalis is one of the most important pests in world soybean crop. The most common intervention is the application of agrochemicals, such as novaluron and glyphosate. Among biological control agents, much attention has been drawn to entomopathogenic fungi, as Metarhizium rileyi. Here, we examined the changes that occur in the immune system (total and differential hemocyte count), secondary effects (caterpillar morphology), and oxidative metabolism after the caterpillars were exposed to M. rileyi, novaluron or glyphosate. M. rileyi was able to induce changes in the width, length, and weight of A. gemmatalis pupae, along with an increased in the number of defense cells. Novaluron prompt changes the insect’s immunity, and glyphosate caused milder immunological effects. However, it caused significant secondary effects including malformations in pupae and adults, and an increase in nitric oxide (NO) levels. Mortality observed when treating insects with novaluron and malformations due to glyphosate treatments did not occur due to oxidative stress. However, when insects were exposed to M. rileyi, we verified significantly increased levels of NO and concluded that these insects died due to oxidative stress. Our data provide evidence that contributes to better understanding the mechanism of herbicide-fungus interaction in the management of Anticarsia gemmatalis.

Transcriptional profiling analysis of susceptible and resistant strains of Anticarsia gemmatalis and their response to Bacillus thuringiensis

Anticarsia gemmatalis is one of the main defoliators of soybean in Brazil. Bacillus thuringiensis (Bt) transgenic crops are used for their management. In this paper we used RNA-seq to explore the response of A. gemmatalis to Bt HD73, as well as to detect transcriptional differences after Bt infection between resistant and susceptible strains. A total of 3853 and 6224 differentially expressed genes (DGEs) were identified in susceptible and resistant larvae after Bt exposure, respectively. We identified 2143 DEGs between susceptible and resistant larvae and 1991 between susceptible and resistant larvae Bt exposed. Immunity-related genes, Bt toxins receptors, proteases, genes involved in metabolic processes, transporters, cuticle proteins and mobile elements have been identified. qRT-PCR data demonstrated upregulation of five genes in susceptible strain after Bt exposure. These results provide insights to understand the molecular and cellular mechanisms of response to Bt that could be used in strategies to control agricultural pests.

Intestinal proteases profiling from Anticarsia gemmatalis and their binding to inhibitors.

Although the importance of intestinal hydrolases is recognized, there is little information on the intestinal proteome of lepidopterans such as Anticarsia gemmatalis. Thus, we carried out the proteomic analysis of the A. gemmatalis intestine to characterize the proteases by LC/MS. We examined the interactions of proteins identified with protease inhibitors (PI) using molecular docking. We found 54 expressed antigens for intestinal protease, suggesting multiple important isoforms. The hydrolytic arsenal featured allows for a more comprehensive understanding of insect feeding. The docking analysis showed that the soybean PI (SKTI) could bind efficiently with the trypsin sequences and, therefore, insect resistance does not seem to involve changing the sequences of the PI binding site. In addition, a SERPIN was identified and the interaction analysis showed the inhibitor binding site is in contact with the catalytic site of trypsin, possibly acting as a regulator. In addition, this SERPIN and the identified PI sequences can be targets for the control of proteolytic activity in the caterpillar intestine and serve as a support for the rational design of a molecule with greater stability, less prone to cleavage by proteases and viable for the control of insect pests such as A. gemmatalis.

Effects of intraguild interactions on Anticarsia gemmatalis and Chrysodeixis includens larval fitness and behavior in soybean.

BACKGROUNDCrop pest management requires an understanding of the complex interactions among species that potentially impact crop yield. In soybean, the velvetbean caterpillar, Anticarsia gemmatalis (Hübner), and the soybean looper, Chrysodeixis includens (Walker), are described as key pests, sharing the same feeding guild. We assessed the intraguild interactions of these species under laboratory conditions. Fitness cost study was conducted to examine the influence of competition on insect development. A video tracking system was used to evaluate behavioral parameters during larval interactions in scenarios with and without food availability.RESULTSIn the fitness cost assay, pupal weight was not significantly affected, regardless of sex. However, larval and pupal survival were influenced by the competition, especially in third versus fifth instar scenarios. We detected 40.00% cannibalism and 46.67% predation when A. gemmatalis and C. includens third instars competed with A. gemmatalis fifth instar, respectively. Distance moved, distance between larvae, body contact (food available) and frequency in food of C. includens larvae were negatively affected by interactions. Anticarsia gemmatalis larvae showed highly active behavior, moving twice or more the distance compared to C. includens larvae, and A. gemmatalis spent more time in body contact with food.CONCLUSIONOur results suggest that A. gemmatalis has a competitive advantage over C. includens. This study provides important information regarding lepidopteran behavior in soybean. We recommended that additional studies are necessary to understand the effects of interactions, especially in field conditions.© 2021 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Azadirachtin-based biopesticide affects the respiration and digestion in Anticarsia gemmatalis caterpillars

Anticarsia gemmatalis (Lepidoptera) is a soybean pest controlled mainly with synthetic insecticides. The effects of azadirachtin-based biopesticide (Azamax®) were investigated against A. gemmatalis caterpillars for toxicity, survival, respiration, antifeeding effects, and midgut histopathology. Concentration–mortality show LC50 = 0.58 g L−1 to A. gemmatalis. Survival caterpillars decrease to 30% after biopesticide exposure. The biopesticide decrease the respiration rate and food consumption by caterpillars. The midgut of insects exposed to the biopesticide undergoes histopathological changes. Azadirachtin-based biopesticide exhibits lethal and sublethal on A. gemmatalis and can be used in integrated pest management programs as an alternative to control this pest.

Molecular characterization of a Bacillus thuringiensis strain from Argentina, toxic against Lepidoptera and Coleoptera, based on its whole-genome and Cry protein analysis

The insecticidal proteins of Bacillus thuringiensis are used in formulations of spore-crystal complexes and their genes have been incorporated into several crops, providing a model for genetic engineering in agriculture. Despite the variability of the Cry proteins described so far, it is still necessary to look for toxins with a broad spectrum of action, since a significant number of pests are not controlled with the available Cry proteins. It is also important to provide alternatives to address the problem of insect resistance, which has already appeared with the use of formulations and with transgenic plants that express cry genes that code for insecticidal proteins. The FCC 7 strain was characterized by the ultrastructural parasporal body under optical and electronic microscopy, and for the detection of Cry8-type proteins by genomic and proteomic approaches. The identity of the strain and the presence of putative toxin encoding genes, and virulence factors analyzed by Illumina Miseq 1500 platform genomic sequencing, was confirmed. The identity of the two Cry8 proteins that make up the parasporal body was confirmed by MALDI-TOF/TOF. To expand knowledge about the insecticidal activity of this strain, we conducted preliminary tests against the cotton boll weevil, Anthonomus grandis. Here we report the characterization of a novel B. thuringiensis isolate native to Argentina (FCC 7) toxic against A. grandis. The strain shows a rounded parasporal body harboring mainly a protein of about 140 kDa and two different types of Cry8 proteins. Through whole-genome sequencing, we identified the presence of two cry8-like crystal protein genes, one vpa-like and two vpb-like genes, and multiple virulence factors, deepening the knowledge of a strain that had already been described as toxic against some lepidopterans and coleopterans, including Spodoptera frugiperda, Anticarsia gemmatalis, Tenebrio molitor and Diabrotica speciosa.

Lemongrass essential oil and its components cause effects on survival, locomotion, ingestion, and histological changes of the midgut in Anticarsia gemmatalis caterpillars

Anticarsia gemmatalis caterpillar is controlled with synthetic insecticides, which cause environmental pollution and insect resistance, and new compounds with low environmental impact, need to be investigated. Toxicity, survival, locomotion, feeding, and midgut histopathology of A. gemmatalis caterpillars exposed to lemongrass essential oil (LEO) and its components (citral and geranyl acetate) were investigated. The compounds are toxic to caterpillars with survival decreasing according to concentration. The compounds decrease insect respiration rate, cause repellency responses and midgut histopathological effects in the digestive and globet cells. These findings suggest that these compounds can be alternatives to chemical synthetic insecticides to control this insect.

Associating Site Characteristics With Distributions of Pestiferous and Predaceous Arthropods in Soybean.

Although site-specific pest management has the potential to decrease control costs and environmental impact associated with traditional pest management tactics, the success of these programs relies on the accurate characterization of arthropod distributions within a crop. Because potential correlation of insect counts with remotely sensed field attribute data could help to decrease the costs associated with and need for fine-scale spatial sampling, we chose to determine which within-field variables would be informative of soybean arthropod counts in an attempt to move toward site-specific pest management in this crop. Two soybean fields were grid-sampled for pestiferous and predaceous arthropods, plant productivity estimates, and abiotic variable characterization in 2017-2018. Negative binomial, zero-inflated models were used to estimate presence and counts of soybean arthropod taxa based on normalized difference vegetation index (NDVI), soybean plant height, soil electrical conductivity (ECa), elevation, and calendar week. Among all variables, calendar week was the most reliable predictor of arthropod counts, as it was a significant predictor for a majority of all taxa. Additionally, counts for a majority of pestiferous taxa were significantly associated with distance from the field edge, elevation, soybean plant height, and NDVI. Although site-specific pest management has the potential for reduced management inputs and increased profitability over conventional management (i.e., whole-field) practices, management zones must first be clearly defined based on the within-field variability for the variables of interest. If site-specific pest management practices are to be applied in soybean, calendar week (and associated soybean phenology), soybean plant height (and associated elevation), and NDVI may be useful for describing the distributions of pests, such as kudzu bug, Megacopta cribraria (Hemiptera: Plataspidae) (Fabricius), green cloverworm, Hypena scabra (Lepidoptera: Erebidae) (Fabricius), velvetbean caterpillar, Anticarsia gemmatalis (Lepidoptera: Erebidae) (Hübner), and soybean looper, Chrysodeixis includens (Lepidoptera: Noctuidae) (Walker).

Current Distribution and Population Persistence of Five Lepidopteran Pests in U.S. Soybean

AbstractThe distribution of lepidopteran pests in soybean (Glycine max (L.) Merr.) is a current knowledge gap limiting accurate prioritization of Integrated Pest Management (IPM) research. Regional characterizations of lepidopteran distribution in soybean are now more than 25 yr old. The goal of this study was to generate a contemporary assessment of the distribution and population persistence of lepidopteran soybean pests. To understand which species are currently infesting soybean and their persistence, we conducted a survey of soybean entomologists with responsibility for approximately 33.6 million hectares of production in 31 U.S. states. Soybean entomologists were asked questions about presence of lepidopteran pests and their persistence in their state. We focused this survey on five lepidopteran pests: corn earworm (Helicoverpa zea Boddie) (Lepidoptera: Noctuidae), green cloverworm (Hypena scabra Fabricius) (Lepidoptera: Erebidae), painted lady (Vanessa cardui L.) (Lepidoptera: Nymphalidae), soybean looper (Chrysodeixis includens Walker) (Lepidoptera: Noctuidae), and velvetbean caterpillar (Anticarsia gemmatalis Hübner) (Lepidoptera: Erebidae). Soybean entomologists also provided insight into regionally relevant or sporadic lepidopteran soybean pests. Participants were also questioned about common scouting practices in each state. Results of this survey highlight dissimilar geographic distribution and relative persistence of lepidopteran pests in soybean. Clear differences in occurrence and abundance among species provide important contemporary distributions and persistence estimates. Assessments of scouting practices demonstrate a need to improve IPM adoption in some states. Results of this study and its complementary pest profile (concurrently published in JIPM) provide a contemporary foundation for studies of lepidopteran soybean pests.

Identification and recombinant expression of an antimicrobial peptide (cecropin B-like) from soybean pest Anticarsia gemmatalis.

BackgroundInsects can be found in numerous diverse environments, being exposed to pathogenic organisms like fungi and bacteria. Once these pathogens cross insect physical barriers, the innate immune system operates through cellular and humoral responses. Antimicrobial peptides are small molecules produced by immune signaling cascades that develop an important and generalist role in insect defenses against a variety of microorganisms. In the present work, a cecropin B-like peptide (AgCecropB) sequence was identified in the velvetbean caterpillar Anticarsia gemmatalis and cloned in a bacterial plasmid vector for further heterologous expression and antimicrobial tests. Methods AgCecropB sequence (without the signal peptide) was cloned in the plasmid vector pET-M30-MBP and expressed in the Escherichia coli BL21(DE3) expression host. Expression was induced with IPTG and a recombinant peptide was purified using two affinity chromatography steps with Histrap column. The purified peptide was submitted to high-resolution mass spectrometry (HRMS) and structural analyses. Antimicrobial tests were performed using gram-positive (Bacillus thuringiensis) and gram-negative (Burkholderia kururiensis and E. coli) bacteria. ResultsAgCecropB was expressed in E. coli BL21 (DE3) at 28°C with IPTG 0.5 mM. The recombinant peptide was purified and enriched after purification steps. HRMS confirmed AgCrecropB molecular mass (4.6 kDa) and circular dichroism assay showed α-helix structure in the presence of SDS. AgCrecropB inhibited almost 50% of gram-positive B. thuringiensis bacteria growth. ConclusionsThe first cecropin B-like peptide was described in A. gemmatalis and a recombinant peptide was expressed using a bacterial platform. Data confirmed tertiary structure as predicted for the cecropin peptide family. AgCecropB was capable to inhibit B. thuringiensis growth in vitro.

Integral Characterization of the 16S rRNA Gene of Non-sporulating Bacteria and Its Action Against Anticarsia gemmatalis Hübner (Lepidoptera: Erebidae)

Brazil is the world’s largest producer of soybean (Glycine max), an extremely important legume due to its source of proteins and essential oils for humans and animals, besides to its applications in the various branches of industry. The velvetbean caterpillar [Anticarsia gemmatalis Hübner (Lepidoptera: Erebidae)] is a great pest that affects this crop and has been controlled by chemical and biological pesticides based on Bacillus thuringiensis. The objectives of this work were to prospect soil microorganisms, to characterize them using the 16S rRNA gene and to perform bioassays to analyze the lethality or subletality of these isolates against A. gemmatalis larvae. The DNA sequencing of the marker gene was complete, covering all conserved regions of it to determine the phylogenetic position of the isolates. Regarding to bioassays, subletality efficacy were low both for sporulant and for the non-sporulant bacterial strains tested. However, based on the signature by complete 16S rRNA analyses of the non-sporulating bacterial isolates, new characteristics worth of studying and prospecting biotechnologically became available.

Effect of the Cry1, Cry2 and Vip3 protein combinations on the control of Anticarsia gemmatalis (Erebidae) and Chrysodeixis includens (Noctuidae) Lepidoptera

The transgenic soybean expressing the cry1Ac insecticidal gene of the Bacillus thuringiensis bacterium has been widely adopted in agriculture. This bacterium acts as an efficient control agent of pest insects due to the expression of different proteins with entomopathogenic activity such as Cry and Vip. This study investigates the toxicity and mode of interaction between the Cry1Ab, Cry1Ac, Cry2Aa, Cry2Ab and Vip3Aa proteins against Chrysodeixis includens and Anticarsia gemmatalis larvae. The combination of the Cry1Ac, Cry2Aa and Vip3Aa toxins had a synergistic effect for C. includens, however, only Vip3Aa and Cry2Aa exhibited synergistic interaction against A. gemmatalis. Cry1Ab and Cry1Ac proteins have been shown to share receptors with the two species tested, however, Cry1Ac does not share binding sites with Cry2Aa, Cry2Ab, and Vip3Aa in both species. Besides helping the resistance management to B. thuringiensis proteins, the combination of Cry and Vip proteins can increase toxicity through synergistic action.

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.

Exposure to chlorantraniliprole reduces locomotion, respiration, and causes histological changes in the midgut of velvetbean caterpillar Anticarsia gemmatalis (Lepidoptera: Noctuidae)

The anthranilic diamide, chlorantraniliprole is a systemic insecticide affecting ryanodine receptors. This insecticide is used to control caterpillars in soybean crops because it has low toxicity to non-target organisms. The objective was to identify side-effects of chlorantraniliprole on midgut histopathology, respiration and behavior of the velvetbean caterpillar Anticarsia gemmatalis in laboratoty. Chlorantraniliprole has LC50 = 0.61 (0.58–0.64) mg mL−1 for A. gemmatalis fourth instar caterpillars after 96 h. The insecticide causes severe histopathological effects in the midgut with epithelial disorganization, microvilli degeneration, cytoplasm vacuolization, cell fragmentation, and peritrophic matrix disorganization. The respiratory rate and the walking speed decrease, whereas the resting period increase for caterpillars exposed to this insecticide. Chlorantraniliprole is toxic to A. gemmatalis at median lethal concentrations causing severe histological and ultrastructural changes with degeneration of the midgut epithelium, reduction of respiratory rates and inducing an arresting behavioral response of this insect.

Inhibitory effects of tripeptides to enzymatic activity and life cycle parameters of Anticarsia gemmatalis

Chemical pesticides are the main tool used to handle lepidopteran insects in agriculture. However, in addition to causing environmental problems, the insects have acquired resistance to these molecules. Thus, the development of environmentally friendly biopesticides with higher specificity, such as organic protease inhibitors, has been encouraged. This work evaluated the inhibitory effects of two tripeptides, Pep1 and Pep2, obtained by removing the cleavage site of tripeptidyl substrates for trypsin-like proteases. Anticarsia gemmatalis were used as biochemical model and the trypsin-like activity of larvae exposed to different dosages of the tripeptides and protease inhibitors merged in an artificial diet was evaluated. Further, we selected one dose of each tripeptide to set a biological assay, where the total-proteolytic, cysteine-proteases, chymotrypsin-like and trypsin-like activities were determined. The tripeptides shown to have potential to inhibit trypsin-like serine proteases of A. gemmatalis midgut, being the lowest activity detected at doses of 170.4 μM and 251.2 μM for Pep1 and Pep2, respectively. The survival curves obtained using Kaplan-Meier estimators indicated that Pep2 and Pep1 affected survival when comparing to the control group. However, the effect of Pep1 on survival was more pronounced indicating the lowest percent of survival at the end of the larval phase (20%). In addition, larvae exposed to Pep1 maintained normal total proteolytic activity at the expense of the activities of chymotrypsin-like and cysteine proteases. Collectively, our findings revealed that the tripeptides, especially Pep1, exhibit toxic effects on A. gemmatalis and should be tested against other lepidopteran insects.

The pest-specific effects of glyphosate on functional response of a wolf spider

Although glyphosate is widely used for weed pest control, it might have negative side effects on natural enemies. Wolf spiders are one of the most representative predators found on soybean crops in Uruguay, preying on a wide variety of potential pests. However, the sublethal effects that pesticides might have on this group have been poorly explored for South American species. Herein, we explored the sublethal effects of glyphosate on the functional response of the wolf spider Hogna cf. bivittata against three potential pest insects, namely ant (Acromyrmex sp.), caterpillar (Anticarsia gemmatalis), and cricket (Miogryllus sp.). We contaminated residually adult females of the species Hogna cf. bivittata with glyphosate (Roundup®) and compared their functional response against non-contaminated spiders. We did not observe any mortality during the study. We found that overall Hogna cf. bivittata showed a functional response type II against crickets and caterpillars but no functional response to ants. Contaminated spiders killed less ants and caterpillars in comparison to the control group, probably as a consequence of the irritating effects of glyphosate. We did not observe differences in functional response to crickets at the evaluated densities, probably as a consequence of the low capture rate against this prey. Although glyphosate does not specifically target spiders, it might have negative sublethal effects on native predators such as Hogna cf. bivittata. Further studies should explore effect of glyphosate on other native predators from South American crops.

Leaf metabolic profiles of two soybean genotypes differentially affect the survival and the digestibility of Anticarsia gemmatalis caterpillars

Insect pests such as Anticarsia gemmatalis cause defoliation and yield losses. Soybean breeding has obtained resistant genotypes, however the mechanism remains unknown. Studies indicated the presence of deterrents compounds in the resistant genotype IAC17, and their leaf metabolite profiles were compared to the susceptible genotype UFV105, which was elicited or not by caterpillar infestation. Cluster analysis indicated a significative distinction between these profiles as well as differences in plant defense pathways. Methylquercetins were constitutively present in the largest concentrations, specifically in the IAC17. Relationship between the resistance and the levels of phytohormones jasmonic acid, abscisic acid and salicylic acid was not observed. However, 1-aminocyclopropane -1carboxylic acid levels indicated that the ethylene may be involved in the constitutive biosynthesis of bioactive compounds. Extracts were added to the diets at three different concentrations to evaluate the effect on caterpillar survival. Lowest survival rates were observed when extracts from the resistant IAC 17 were used, at the lowest concentrations. Survival rates were not higher when IAC 17 infested by caterpillars were used. On the other hand, when extracts from the susceptible were used, the survival reductions were only observed in the highest extract concentrations. These supplementations of the diet reduced the digestive capacity, agreeing with the proteolytic activities, whereas malformations of the intestinal cells were dose dependent. The inhibitory effects persisted in higher dilutions only for the IAC17. Constitutive resistance was also explained by higher levels of protease inhibition. These results can be useful to elucidate the genes and cascades controlling the resistance.

Side-effects caused by chlorpyrifos in the velvetbean caterpillar Anticarsia gemmatalis (Lepidoptera: Noctuidae)

Anticarsia gemmatalis is pest in soybean crops, which main control method is the use of insecticides. Toxicity, survival and side-effects (locomotor behavior, respiration, food consumption and histopathology of the midgut) of A. gemmatalis exposed to chlorpyrifos via ingestion were evaluated. Chlorpyrifos was toxic to A. gemmatalis (LC50 = 0.58 g L−1 and LC90 = 0.85 g L−1) and survival decrease from 99% in control to 30% in insects exposed to LC50 chlorpyrifos. The insecticide decreased the respiration rate, food consumption, altered behavioral responses with hyperactivity in A. gemmatalis. In addition the midgut had histopathological changes characterized by damages in the peritrophic matrix and striated border, high cytoplasm vacuolization, cell fragments released in the lumen and apocrine secretion, which can compromise the digestion of this insect. Chlorpyrifos is toxic to A. gemmatalis larvae, affecting survival, locomotion, respiration, food consumption and the midgut.

Efficacy of an oil‐based formulation combining Metarhizium rileyi and nucleopolyhedroviruses against lepidopteran pests of soybean

AbstractA biopesticide that combines Metarhizium rileyi and nucleopolyhedrovirus (NPV) in an oil dispersion (OD) was developed. Its efficacy against Anticarsia gemmatalis and Chrysodeixis includens was evaluated under laboratory and field conditions. First, each of the two selected fungal strains, combined or not in OD preparations with AgMNPV or ChinNPV, was tested in the laboratory against A. gemmatalis and C. includens, respectively. AgMNPV–fungus combinations had similar effect on overall mortality of A. gemmatalis compared to each pathogen used alone. C. includens was less susceptible to infection by M. rileyi strains, and ChinNPV became the major cause of larval death. The OD formulations with M. rileyi and/or NPV were applied in infested soybean fields. ChinNPV + M. rileyi caused 50.8% of overall mortality six days after spraying in a soybean crop infested by both pests, whereas for treatments with the fungus or the virus alone the percentages were 41.2% and 8.8%, respectively. In another field infested solely by A. gemmatalis, insect mortality was similar for the AgMNPV–fungus combination and the virus used alone. Larval parasitism by wasps and dipterans was not affected following biopesticide spraying. Our results suggest that the performance of a dual‐action biopesticide is related to the NPV and fungal strains that are combined and the proportion of host populations simultaneously infesting the crop.

Noncompetitive tight-binding inhibition of Anticarsia gemmatalis trypsins by Adenanthera pavonina protease inhibitor affects larvae survival.

The economic loss in soybean crops caused by the Lepidoptera insects has encouraged the search for new strategies to control this pest, which are currently based on synthetic insecticides. This paper evaluated the ability of ApTI (Adenanthera pavonina trypsin inhibitor) to inhibit trypsin-like proteins from Anticarsia gemmatalis by docking, molecular dynamics, and enzymatic and survival assay. The docking and molecular dynamic simulation between trypsin and ApTI were performed using the program CLUSPRO and NAMD, respectively. The inhibitory constant Ki and the inhibition type were determined through chromogenic assays. The survival assay of neonatal larvae under treatment with artificial diet supplemented with ApTI was also performed. The ApTI binding site was predicted to block substrate access to trypsin due to four interactions with the enzyme, producing a complex with a surface area of 1,183.7 Å2 . The kinetic analysis revealed a noncompetitive tight-binding mechanism. The survival curves obtained using Kaplan-Meier estimators indicated that the highest larvae mortality was 60%, using 1.2 mg of ApTI per 100 ml of artificial diet. The in vitro, in vivo, and in silico studies demonstrated that ApTI is a strong noncompetitive inhibitor of trypsin with biotechnological potential for the control of A. gemmatalis insect.