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Although the boll weevil (BW), Anthonomus grandis grandis (Coleoptera: Curculionidae) has been attributed to the significant losses caused to cotton yield in the Americas, the categorization as a quarentenary pest in places where it is still not occurring has increased its relevance worldwide. In areas where it is widespread, pest suppression relies on many broad-spectrum insecticide applications. However, other control tactics are sought. Considering that early-flowering cultivars escape from boll weevil infestation, we investigated if three different planting dates (November, December, and January) could alter the plant life cycle, allowing the plants to escape from boll weevil infestation. Field trials were run in two seasons (2014/2015 and 2017/2018), and variables (days required to reach each flowering stage, fruiting plant structures-undamaged and damaged by the BW, and totals-number of boll weevils on plants and that had emerged from fallen structures on the ground) were assessed over 29 and 33 weeks, respectively. Based on the number of days required to initiate and terminate the flowering stage, the time to reach the economic threshold (ET), the number of undamaged, damaged, and the total reproductive structures, we concluded that planting dates in December for the Central Cerrado of Brazil should be preferred over the other two tested dates. Cultivations run at this planting date, anticipating the flowering period initiation and termination, reduced infested flowering structures, and delayed the decision making to control the pest, when compared to the other two planting dates.

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Anthonomus grandis Boheman (Coleoptera: Curculionidae) survives on alternative diets; however, this induces reproductive diapause. The objective was to evaluate the morphology and morphometry of the reproductive tract of this weevil after feeding on alternative diets. The experimental design was completely randomized with 160 replications and treatments arranged in a factorial design 3 × 3, represented by A. grandis adults fed on 3 food types (fragments of banana (T1) or orange (T2) endocarp, or with cotton squares of cultivar BRS 286 (T3, control)) and three evaluation periods (30, 60, and 90 days) and after each of these periods they were fed with cotton squares for 10 days. The reproductive tract of 100% of A. grandis females fed banana endocarp, orange endocarp, and cotton squares for 30 and 60 days and then cotton squares were morphologically adequate for reproduction, and after 90 days, only 50% of those fed cotton squares were in this condition. The length of the ovarioles and the width of the mature oocyte were greater for A. grandis fed on cotton squares and smaller in those with banana and orange endocarps. Histological sections reveal that male testes even with strong degenerative signals are already producing spermatozoa. On the other hand, females displayed ovaries with nurse cells in the tropharium and some maturing oocytes in the vitellarium. The body length was longer and the testis area and diameter smaller in males fed on cotton squares than in those with banana and orange endocarp. Anthonomus grandis females fed for ≥90 days with alternative food sources do not recover the functionality of their reproductive tract, even after consuming, for 10 days, a diet that favors reproduction. On the other hand, the males remain with their reproductive organs functional with this condition.

RESUMEN

The cotton boll weevil (CBW) (Anthonomus grandis) is one of the major insect pests of cotton in Brazil. Currently, CBW control is mainly achieved by insecticide application, which is costly and insufficient to ensure effective crop protection. RNA interference (RNAi) has been used in gene function analysis and the development of insect control methods. However, some insect species respond poorly to RNAi, limiting the widespread application of this approach. Therefore, nanoparticles have been explored as an option to increase RNAi efficiency in recalcitrant insects. Herein, we investigated the potential of chitosan-tripolyphosphate (CS-TPP) and polyethylenimine (PEI) nanoparticles as a dsRNA carrier system to improve RNAi efficiency in the CBW. Different formulations of the nanoparticles with dsRNAs targeting genes associated with juvenile hormone metabolism, such as juvenile hormone diol kinase (JHDK), juvenile hormone epoxide hydrolase (JHEH), and methyl farnesoate hydrolase (MFE), were tested. The formulations were delivered to CBW larvae through injection (0.05-2 µg), and the expression of the target genes was evaluated using RT-qPCR. PEI nanoparticles increased targeted gene silencing compared with naked dsRNAs (up to 80%), whereas CS-TPP-dsRNA nanoparticles decreased gene silencing (0%-20%) or led to the same level of gene silencing as the naked dsRNAs (up to 50%). We next evaluated the effects of targeting a single gene or simultaneously targeting two genes via the injection of naked dsRNAs or dsRNAs complexed with PEI (500 ng) on CBW survival and phenotypes. Overall, the gene expression analysis showed that the treatments with PEI targeting either a single gene or multiple genes induced greater gene silencing than naked dsRNA (∼60%). In addition, the injection of dsJHEH/JHDK, either naked or complexed with PEI, significantly affected CBW survival (18% for PEI nanoparticles and 47% for naked dsRNA) and metamorphosis. Phenotypic alterations, such as uncompleted pupation or malformed pupae, suggested that JHEH and JHDK are involved in developmental regulation. Moreover, CBW larvae treated with dsJHEH/JHDK + PEI (1,000 ng/g) exhibited significantly lower survival rate (55%) than those that were fed the same combination of naked dsRNAs (30%). Our findings demonstrated that PEI nanoparticles can be used as an effective tool for evaluating the biological role of target genes in the CBW as they increase the RNAi response.

RESUMEN

The boll weevil, Anthonomus grandis grandis Boheman, is one of the most historically impactful insects due to its near destruction of the US cotton industry in the early 20th century. Contemporary efforts to manage this insect primarily use pheromone baited traps for detection and organophosphate insecticides for control, but this strategy is not sustainable due to financial and environmental costs. We present a high-quality boll weevil genome assembly, consisting of 306 scaffolds with approximately 24,000 annotated genes, as a first step in the identification of gene targets for novel pest control. Gene content and transposable element distribution are similar to those found in other Curculionidae genomes; however, this is the most contiguous and only assembly reported to date for a member in the species-rich genus Anthonomus. Transcriptome profiles across larval, pupal, and adult life stages led to identification of several genes and gene families that could present targets for novel control strategies.

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Abstract The cotton boll weevil, Anthonomus grandis grandis Boheman (Coleoptera: Curculionidae), is a key cotton crop pest in Brazil. Adverse climatic factors, such as high temperatures and low soil moisture, dehydrate oviposited cotton squares (bud flowers) on the ground and cause high mortality of its offspring within these plant structures. The objective of this research was to evaluate the mortality of the cotton boll weevil in drip and sprinkler irrigated cotton crops. The experimental design was in randomized blocks with two treatments: drip (T1) and sprinkler (T2, control) irrigated cotton crops with sixteen replications. Each parcel had one emergence cage, installed between two cotton rows per irrigation system, with 37 cotton squares with opened oviposition punctures and yellowish bracts, to capture adult cotton boll weevils. The average number of boll weevils that emerged from the cotton squares and the causes of mortality at different development stages were determined per treatment. Third-generation life tables of the boll weevil were prepared using the natural mortality data in drip and sprinkler irrigation treatments and plus actual, apparent and indispensable mortality rates and the lethality of each mortality cause. We conclude that the application of water directly to the root zone of the plants in a targeted manner, using the drip irrigation system, can cause high mortality of the cotton boll weevil immature stages inside cotton squares fallen on the ground. This is because the cotton squares fallen on the drier and hotter soil between the rows of drip-irrigated cotton dehydrates causing the boll weevils to die. This is important because it can reduce its population density of the pest and, consequently, the number of applications of chemical insecticides for its control. Thus, contributing to increase the viability of cotton production, mainly in areas of the Brazilian semiarid region where the cotton is cultivated in organic system.

Resumo O bicudo-do-algodoeiro, Anthonomus grandis grandis Boheman (Coleoptera: Curculionidae), é uma praga-chave da cultura do algodão no Brasil. Fatores climáticos adversos, como altas temperaturas e baixa umidade do solo, desidratam os botões florais de algodão ovipositados caídos ao solo e causam alta mortalidade de seus descendentes dentro dessas estruturas vegetais.O objetivo desta pesquisa foi avaliar a mortalidade do bicudo-do-algodoeiro em lavouras de algodão irrigadas por gotejamento e aspersão. O delineamento experimental foi em blocos casualizados com dois tratamentos: cultivo de algodão irrigado por gotejamento (T1) e aspersão (T2, testemunha) com dezesseis repetições. Cada parcela possuía uma gaiola de emergência, instalada entre duas fileiras de algodão por sistema de irrigação, com 37 botões florais de algodão com orifícios de oviposição, brácteas abertas e amarelecidas, para captura do bicudo adulto. O número médio de bicudos que emergiu dos botões florais de algodão e as causas de mortalidade, em diferentes estágios de desenvolvimento, foram determinados por tratamento. As tabelas de vida de terceira geração do bicudo-do-algodoeiro foram preparadas usando os dados de mortalidade natural em tratamentos de irrigação por gotejamento e aspersão e as taxas de mortalidade reais, aparentes e indispensáveis e a letalidade de cada causa de mortalidade. Concluímos que a aplicação de água diretamente na zona radicular das plantas de forma direcionada, utilizando o sistema de irrigação por gotejamento, pode causar altas mortalidade dos estágios imaturos do bicudo-do-algodoeiro nos botões florais de algodão caídos no solo. Isso ocorre porque os botões florais de algodão caídos no solo mais seco e mais quente entre as fileiras do algodão irrigado por gotejamento desidratam, causando a morte dos bicudos. Isso é importante por poder reduzir a densidade populacional desse inseto e, consequentemente, o número de aplicações de inseticidas químicos para seu controle. Assim, contribuindo para aumentar a viabilidade da produção de algodão, principalmente em áreas do semiárido brasileiro em sistema orgânico.

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The cotton boll weevil, Anthonomus grandis grandis Boheman (Coleoptera: Curculionidae), is a key cotton crop pest in Brazil. Adverse climatic factors, such as high temperatures and low soil moisture, dehydrate oviposited cotton squares (bud flowers) on the ground and cause high mortality of its offspring within these plant structures. The objective of this research was to evaluate the mortality of the cotton boll weevil in drip and sprinkler irrigated cotton crops. The experimental was in included randomized blocks with two treatments: drip (T1) and sprinkler (T2, control) irrigated cotton crops with sixteen replications. Each parcel had one emergence cage, installed between two cotton rows per irrigation system, with 37 cotton squares with opened oviposition punctures and yellowish bracts, to capture adult cotton boll weevils. The average number of boll weevils that emerged from the cotton squares and the causes of mortality at different development stages were determined per treatment. Third-generation life tables of the boll weevil were prepared using the natural mortality data in drip and sprinkler irrigation treatments and plus actual, apparent and indispensable mortality rates and the lethality of each mortality cause. We conclude that the application of water directly to the root zone of the plants in a targeted manner, using the drip irrigation system, can cause high mortality of the cotton boll weevil immature stages inside cotton squares fallen on the ground. This is because the cotton squares fallen on the drier and hotter soil between the rows of drip-irrigated cotton dehydrates causing the boll weevils to die. This is important because it can reduce its population density of the pest and, consequently, the number of applications of chemical insecticides for its control. Thus, contributing to increase the viability of cotton production, mainly in areas of the Brazilian semiarid region where the cotton is cultivated in organic system.

O bicudo-do-algodoeiro, Anthonomus grandis grandis Boheman (Coleoptera: Curculionidae), é uma praga-chave da cultura do algodão no Brasil. Fatores climáticos adversos, como altas temperaturas e baixa umidade do solo, desidratam os botões florais de algodão ovipositados caídos ao solo e causam alta mortalidade de seus descendentes dentro dessas estruturas vegetais. O objetivo desta pesquisa foi avaliar a mortalidade do bicudo-do-algodoeiro em lavouras de algodão irrigadas por gotejamento e aspersão. O delineamento experimental foi em blocos casualizados com dois tratamentos: cultivo de algodão irrigado por gotejamento (T1) e aspersão (T2, testemunha) com dezesseis repetições. Cada parcela possuía uma gaiola de emergência, instalada entre duas fileiras de algodão por sistema de irrigação, com 37 botões florais de algodão com orifícios de oviposição, brácteas abertas e amarelecidas, para captura do bicudo adulto. O número médio de bicudos que emergiu dos botões florais de algodão e as causas de mortalidade, em diferentes estágios de desenvolvimento, foram determinados por tratamento. As tabelas de vida de terceira geração do bicudo-do-algodoeiro foram preparadas usando os dados de mortalidade natural em tratamentos de irrigação por gotejamento e aspersão e as taxas de mortalidade reais, aparentes e indispensáveis e a letalidade de cada causa de mortalidade. Concluímos que a aplicação de água diretamente na zona radicular das plantas de forma direcionada, utilizando o sistema de irrigação por gotejamento, pode causar altas mortalidade dos estágios imaturos do bicudo-do-algodoeiro nos botões florais de algodão caídos no solo. Isso ocorre porque os botões florais de algodão caídos no solo mais seco e mais quente entre as fileiras do algodão irrigado por gotejamento desidratam, causando a morte dos bicudos. Isso é importante por poder reduzir a densidade populacional desse inseto e, consequentemente, o número [...].

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Abstract The cotton boll weevil, Anthonomus grandis grandis Boheman (Coleoptera: Curculionidae), is a key cotton crop pest in Brazil. Adverse climatic factors, such as high temperatures and low soil moisture, dehydrate oviposited cotton squares (bud flowers) on the ground and cause high mortality of its offspring within these plant structures. The objective of this research was to evaluate the mortality of the cotton boll weevil in drip and sprinkler irrigated cotton crops. The experimental design was in randomized blocks with two treatments: drip (T1) and sprinkler (T2, control) irrigated cotton crops with sixteen replications. Each parcel had one emergence cage, installed between two cotton rows per irrigation system, with 37 cotton squares with opened oviposition punctures and yellowish bracts, to capture adult cotton boll weevils. The average number of boll weevils that emerged from the cotton squares and the causes of mortality at different development stages were determined per treatment. Third-generation life tables of the boll weevil were prepared using the natural mortality data in drip and sprinkler irrigation treatments and plus actual, apparent and indispensable mortality rates and the lethality of each mortality cause. We conclude that the application of water directly to the root zone of the plants in a targeted manner, using the drip irrigation system, can cause high mortality of the cotton boll weevil immature stages inside cotton squares fallen on the ground. This is because the cotton squares fallen on the drier and hotter soil between the rows of drip-irrigated cotton dehydrates causing the boll weevils to die. This is important because it can reduce its population density of the pest and, consequently, the number of applications of chemical insecticides for its control. Thus, contributing to increase the viability of cotton production, mainly in areas of the Brazilian semiarid region where the cotton is cultivated in organic system.

Resumo O bicudo-do-algodoeiro, Anthonomus grandis grandis Boheman (Coleoptera: Curculionidae), é uma praga-chave da cultura do algodão no Brasil. Fatores climáticos adversos, como altas temperaturas e baixa umidade do solo, desidratam os botões florais de algodão ovipositados caídos ao solo e causam alta mortalidade de seus descendentes dentro dessas estruturas vegetais.O objetivo desta pesquisa foi avaliar a mortalidade do bicudo-do-algodoeiro em lavouras de algodão irrigadas por gotejamento e aspersão. O delineamento experimental foi em blocos casualizados com dois tratamentos: cultivo de algodão irrigado por gotejamento (T1) e aspersão (T2, testemunha) com dezesseis repetições. Cada parcela possuía uma gaiola de emergência, instalada entre duas fileiras de algodão por sistema de irrigação, com 37 botões florais de algodão com orifícios de oviposição, brácteas abertas e amarelecidas, para captura do bicudo adulto. O número médio de bicudos que emergiu dos botões florais de algodão e as causas de mortalidade, em diferentes estágios de desenvolvimento, foram determinados por tratamento. As tabelas de vida de terceira geração do bicudo-do-algodoeiro foram preparadas usando os dados de mortalidade natural em tratamentos de irrigação por gotejamento e aspersão e as taxas de mortalidade reais, aparentes e indispensáveis e a letalidade de cada causa de mortalidade. Concluímos que a aplicação de água diretamente na zona radicular das plantas de forma direcionada, utilizando o sistema de irrigação por gotejamento, pode causar altas mortalidade dos estágios imaturos do bicudo-do-algodoeiro nos botões florais de algodão caídos no solo. Isso ocorre porque os botões florais de algodão caídos no solo mais seco e mais quente entre as fileiras do algodão irrigado por gotejamento desidratam, causando a morte dos bicudos. Isso é importante por poder reduzir a densidade populacional desse inseto e, consequentemente, o número de aplicações de inseticidas químicos para seu controle. Assim, contribuindo para aumentar a viabilidade da produção de algodão, principalmente em áreas do semiárido brasileiro em sistema orgânico.

RESUMEN

Cotton is the most important crop for fiber production worldwide. However, the cotton boll weevil (CBW) is an insect pest that causes significant economic losses in infested areas. Current control methods are costly, inefficient, and environmentally hazardous. Herein, we generated transgenic cotton lines expressing double-stranded RNA (dsRNA) molecules to trigger RNA interference-mediated gene silencing in CBW. Thus, we targeted three essential genes coding for chitin synthase 2, vitellogenin, and ecdysis-triggering hormone receptor. The stability of expressed dsRNAs was improved by designing a structured RNA based on a viroid genome architecture. We transformed cotton embryos by inserting a promoter-driven expression cassette that overexpressed the dsRNA into flower buds. The transgenic cotton plants were characterized, and positive PCR transformed events were detected with an average heritability of 80%. Expression of dsRNAs was confirmed in floral buds by RT-qPCR, and the T1 cotton plant generation was challenged with fertilized CBW females. After 30 days, data showed high mortality (around 70%) in oviposited yolks. In adult insects fed on transgenic lines, chitin synthase II and vitellogenin showed reduced expression in larvae and adults, respectively. Developmental delays and abnormalities were also observed in these individuals. Our data remark on the potential of transgenic cotton based on a viroid-structured dsRNA to control CBW.

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Growers rely on synthetic insecticides to control the boll weevil throughout the reproductive cotton stage. An average of 19.6 insecticide applications (range: 10 to 30) for control of boll weevil were found in a survey with growers in the Brazilian Cerrado, covering an area of 494,100 hectares of cotton. Twenty-one insecticides were applied, with 64.8% of the applications made with malathion, fipronil, carbosulfan, and thiamethoxam + lambda-cyhalothrin. These four insecticides were used by 100, 76, 70, and 62% of the growers, with respectively 7.2, 2.1, 1.8, and 1.6 applications. Growers classified their boll weevil control achieved into four categories (fair, good, very good, or excellent), without correlation between these categories with the number of insecticide applications. Control of cotton regrowth and volunteer cotton plants were the major obstacles for effective boll weevil management, followed by the low efficacy of insecticides. Five registered insecticides to spray cotton against other pests than boll weevil were enlisted by growers with potential for recommendation. A boll weevil standard population for susceptibility was assayed with 27 insecticides and the results presented within a failure risk quotient (FRQ). The FRQ of eight, six, and 13 of the 27 tested insecticides was high, intermediate, and low, respectively. The high FRQ included 7 of 10 pyrethroid formulations, pymetrozine, and methomyl. On the opposite end, fipronil had the lowest FRQ value.

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Synthetic insecticide application is one tactic for reducing boll weevil, Anthonomus grandis grandis Boheman (Coleoptera: Curculionidae), infestations during the cotton, Gossypium hirsutum L., reproductive stage. We assessed the susceptibility of the boll weevil and its natural enemies to ethiprole (mode of action 2B), a phenylpyrazole insecticide, and diagnostic concentrations of ethiprole indicative of boll weevil susceptibility. Differences in the lethal concentrations of ethiprole were calculated with susceptibility ratios based on LC50 ranging from 2.89- to 10.34-fold relative to a natural susceptible population. The lowest and the highest recommended field rates of ethiprole, 100 and 200 g a.i./ha, produced residues that caused 83.3% and 93.7% mortality of weevils caged with cotton leaves from field-treated plants for 8 d. We found that ethiprole was less toxic than fipronil to the boll weevil parasitoid Bracon vulgaris Ashmead (Hymenoptera: Braconidae) and to the red imported fire ant, Solenopsis invicta Buren (Hymenoptera: Formicidae), while fipronil was highly toxic to both. Adult earwigs, Euborellia annulipes Lucas (Dermaptera: Anisolabididae), were relatively tolerant to ethiprole and fipronil at the highest field rates. Pooled LC50-and LC95-concentrations of ethiprole calculated from studied populations were used as diagnostic for boll weevil mortality, and the outcome fitted to the expected mortality for boll weevil populations from different locations serving for further control failure assessment. Ethiprole appears to be suitable for boll weevil control with low impact on natural enemy communities.

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BACKGROUND: Insecticide resistance in arthropods is an inherited trait that has become a major cause of insect pest control failure. Monitoring the level of susceptibility and characterization of the type of resistance of key pest species aims to determine the risk of resistance selection in time to take action to mitigate control failures. Seven populations of the boll weevil, Anthonomus grandis grandis, collected from cotton fields in the Semiarid and Cerrado areas of Brazil, were screened for their resistance to malathion and beta-cyfluthrin, insecticides widely recommended for control of boll weevil and other pests. RESULTS: The levels of adult mortality were variable for beta-cyfluthrin (0-82%) but invariant (100%) for malathion. Bioassays of concentration-mortality were used to determine lethal concentrations (LCs) for each insecticide. The LC-values corroborate the lack of resistance to field rates of malathion but high levels of resistance to beta-cyfluthrin from 62.7- to 439.7-fold. Weevils resistant to beta-cyfluthrin were found through genome sequencing to possess a kdr mutation through the L1014F substitution in the voltage gated-sodium channel gene. CONCLUSIONS: This study found boll weevil resistance to beta-cyfluthrin to be not mediated by carboxylesterases, but with cross-resistance to DDT and carbaryl, and kdr mutation as the major mechanism of the resistance in our samples. Caution is recommended in further use of beta-cyfluthrin against boll weevil due to potential resistance. Monitoring studies using other boll weevil populations are recommended to determine the geographic pattern and extent of pyrethroid resistance. © 2021 Society of Chemical Industry.

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The "cotton boll weevil" (Anthonomus grandis Boheman) is a key pest in America whose larval stage develops within the cotton flower bud. During its development, the larva uses the flower bud as food and as a shelter from predators. This behavior limits the effective control through conventional insecticide applications and biocontrol techniques. Increasing genetic information from insects has allowed the development of new control technologies based on the use of RNA interference (RNAi) to design orally delivered double-stranded RNA (dsRNA) strategies. In this study, we evaluated the effect of continuous oral administration of six specific dsRNA in order to identify an effective target gene for RNAi-mediated control of cotton boll weevil. First, six selected A. grandis gene fragments were amplified and cloned to perform in vivo synthesis of the specific dsRNA, and subsequently, larvae and adults were fed with this dsRNA for 2 weeks. Larvae mortality ranged from 40 to 60% depending on the targeted gene sequence. Indeed, α-amylase and cytochrome p450 dsRNAs were the most effective. Oral administration in adults caused smaller but still significant death rates (15-30%). Thus, the results demonstrated RNAi responses depend on life stages and target genes. The dsRNA ingestion was capable of providing knockdown mRNA levels in cotton boll weevil midgut and this effect was significantly higher in the larval stage. In this study, we present a new report of silencing of midgut genes in A. grandis larva induced by continuously feeding with dsRNA. This potential new tool should be further evaluated in cotton boll weevil control strategies.

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BACKGROUND: Immature stages of boll weevil complete development endophytically leaving only the adult stage accessible for chemical control. We tested the hypothesis that boll weevil colonization of the cotton plants significantly affects their exposure to sprayed insecticides. We determined the adult dispersal toward and within cotton plants, lethal time (LT), and residual control by recommended insecticides (malathion, carbosulfan, thiamethoxam, fipronil, beta-cyfluthrin, lambda-cyhalothrin, and thiamethoxam + lambda-cyhalothrin) through dried residue exposure and residual control regarding the leaf position in the upper and the lower thirds of the treated plant canopy. RESULTS: Newly emerged adults from fallen buds reached the cotton plants by walking (80%) and most of the time settling on cotton bolls in the lower part of the plants (78%). Irrespective of sex and mating status, adults released on the upper part of the plant remained longer on the same release site than the lower part, with some individuals remaining up to 50 h on the same flower bud. The shortest LT90 was found with thiamethoxam (106 h). Fipronil and malathion, respectively, provided the longest (>144 h) and shortest (24 h) residual control times and caused boll weevil mortality above 80%. CONCLUSIONS: These findings suggest that weevils accessing the cotton plants exhibited within-plant distribution that minimizes their contact with insecticide residue on plant foliage. Furthermore, short residual control with malathion, the most used insecticide against boll weevil, and the low susceptibility exhibited by the tested population to pyrethroids highlight the current challenges faced for boll weevil control in Brazilian cotton fields. © 2020 Society of Chemical Industry.

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MAIN CONCLUSION: Cotton genotypes displayed similar volatile organic compound (VOC) profiles, but major differences in terpenoid aldehyde (TA) content. The differences in VOC production were minor among genotypes, but these differences are crucial for boll weevil attraction. Weevils did not display any preference in feeding behaviour towards cotton genotypes, suggesting physiological adaptation to cope with cotton chemical defence mechanisms. Plant cultivar selection for resistance to herbivore pests is an effective, environmentally safe and inexpensive method to implement in integrated pest management programmes. In this study, we evaluated seven cotton genotypes with respect to the production of volatile organic compounds (VOCs) and non-volatile compounds [terpenoid aldehydes (TAs)], and the attraction and feeding preference of adult boll weevils. Chemical analyses of VOCs from BRS-293, BRS-Rubi, CNPA TB-15, CNPA TB-85, CNPA TB-90, Delta Opal, and Empire Glandless showed that there were few qualitative and quantitative differences across the range of genotypes. In contrast, major differences in TA content were observed, with CNPA TB-15 and CNPA TB-85 producing higher levels of TAs compared to the other genotypes. Our results showed that boll weevil attraction to cotton genotypes varied, suggesting that the ratios and quantities of emitted cotton VOCs are important for host location. However, boll weevil feeding behaviour was neither positively nor negatively influenced by the terpenoid content (non-volatile compounds) of cotton genotypes. The results in this study suggest that boll weevils have adapted physiologically to cope with cotton chemical defence mechanisms.

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The departure and arrival of boll weevil in cotton fields have fostered major control decisions against this pest over time. Field colonization and distribution of boll weevil were evaluated using cotton fields from 93.7 to 154 ha each, located in Serra da Petrovina and Campo Verde, both in the Cerrado biome of Mato Grosso State, Brazil, as a function of cotton field bordering vegetation and crop phenology. The monitoring of adult weevils was carried out using traps containing sex and aggregation pheromone. The traps were set up considering the four coordinates of the field, bordering vegetation, and distances from the field margin. Six traps were set in each direction with three representing the field margin (0 m, 60 m, and 120 m from the borderline) and the other three set up equidistant from the center of the field (> 200 m). Traps were evaluated weekly beginning 10 days after plant emergence (DAE) throughout the cotton phenology until harvesting. Boll weevils were caught in the first evaluation 10 DAE, irrespective of the vegetation bordering the cotton field and distance of trap from field margin. The average weevils per trap was relatively low throughout the crop development but increased significantly through the maturation and harvesting periods. Furthermore, the bordering vegetation was not the only determinant factor for boll weevil colonization and distribution throughout the cotton field. Based on these data, boll weevil exhibited early colonization, already occurring beyond the border of the field.

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The cotton boll weevil, Anthonomus grandis, is a major pest of cotton crops in South America. In this work, partial biochemical characterizations of (hemi) cellulases and pectinases activities in the digestive system (head- and gut- extracts) of A. grandis were evaluated. Gut extract section from third instar larvae exhibited endoglucanase, xylanase, ß-glucosidase, and pectinase activities. The endoglucanase and xylanase activities were localized in the foregut, whereas ß-glucosidase activity was mainly detected in the hindgut. In addition, no difference in pectinase activity was observed across the gut sections. Thus, A. grandis digestive system is a potentially interesting reservoir for further lignocellulolytic enzymes research.

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The objective was to register the occurrence of Proctolaelaps bickleyi (Bram) (Acari: Mesostigmata: Melicharidae) in association with adults of the Anthonomus grandis Boheman (Coleoptera: Curculionidae) and to describe aspects of its behavior. This is the first record of a mesostigmatid mite associated with the cotton boll weevil in the Americas. Beetles carrying the mites had lower mobility than usual. The stress caused by this mite attached to the A. grandis body can reduce mating and oviposition of this beetle.

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Boll weevil is the major cotton pest in Brazil, and insecticides are widely recommended against it. We determined the susceptibility of boll weevil to insecticides either in single or in mixture ready-to-use formulations, which are registered to spray cotton fields under the hypothesis that mixtures are more toxic to the target pest. Concentration-mortality curves were determined to adult species, simultaneously through dried residues and ingestion. Ten insecticide formulations were studied with five in mixture (lambda-cyhalothrin + thiamethoxam, lambda-cyhalothrin + chlorantraniliprole, thiamethoxam + chlorantraniliprole, and fenitrothion + esfenvalerate) and their five respective single formulations. Cotton leaf discs and cotyledons were dipped into insecticide dilutions prepared by diluting the commercial products into distilled water. Adult mortality was assessed 48 hours after caging adults on treated and untreated materials. The LC50s-concentrations varied from 0.004 to 0.114 g a.i./L, with a relative potency between single and mixture ones, varying from 1.37- to 29.59-fold. Furthermore, lambda-cyhalothrin and thiamethoxam in single formulation were the most toxic insecticides to boll weevil. Among insecticide mixtures, only lambda-cyhalothrin + chlorantraniliprole resulted in a synergic effect; whereas the remaining mixtures showed an antagonistic effect. Therefore, except for the mixture of lambda-cyhalothrin + chlorantraniliprole, the remaining mixtures did not enhance toxicity against the boll weevil and should be recommended only when aimed at different purposes.(AU)

Bicudo-do-algodoeiro é a principal praga do algodoeiro no Brasil, sendo o uso de inseticidas amplamente recomendado para o seu controle. A suscetibilidade do bicudo-do-algodoeiro foi determinada a inseticidas em formulação simples ou em misturas prontas para uso, as quais têm sido recomendadas para pulverizar campos de algodão sob a hipótese de serem mais tóxicas à praga alvo. Assim, curvas de concentração-mortalidade foram determinadas para adultos do bicudo contaminados, simultaneamente, via resíduo seco e ingestão dos inseticidas. Dez formulações foram estudadas, sendo cinco misturas (lambda-cialotrina + tiametoxam, lambda-cialotrina + clorantraniliprole, tiametoxam + clorantraniliprole e fenitrotiona + esfenvalerato) e suas respectivas cinco formulações simples. Folhas e cotilédones do algodoeiro foram mergulhados em diluições do inseticida preparadas com os produtos comerciais e água destilada. A mortalidade adulta foi avaliada 48 horas após o acondicionamento dos adultos em materiais tratados e não tratados. As concentrações de CL50s variaram de 0,004 a 0,114 g i.a./L, com potência relativa entre formulação simples e misturas, variando de 1,37 a 29,59 vezes. A lambda-cialotrina e o tiametoxam em formulações simples foram os inseticidas mais tóxicos para o bicudo. Entre as misturas, aquela preparada com lambda - cialotrina + clorantraniliprole resultou em um efeito sinérgico, enquanto as demais misturas mostraram um efeito antagonista. Portanto, exceto pela mistura de lambda-cialotrina + clorantraniliprole, as demais misturas não demonstraram maior toxicidade para o bicudo-do-algodoeiro e devem ser recomendadas somente quando objetivam finalidades diferentes.(AU)

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RESUMEN

Preliminary bioassays with whole cultures (WC) of 124 Bacillus thuringiensis strains were performed with neonate larvae of Anthonomus grandis, a major cotton pest in Argentina and other regions of the Americas. Three exotic and four native strains were selected for causing more than 50% mortality. All of them were β-exotoxin producers. The native strains shared similar morphology of parasporal crystals, similar protein pattern and identical insecticidal gene profiles. These features resembled Lepidoptera-toxic strains. Furthermore, these strains showed a Rep-PCR pattern identical to lepidoptericidal strain HD-1, suggesting that these strains may belong to serovar kurstaki. However, some differences were observed in the plasmid profiles and in the production of β-exotoxin. To determine the culture fractions where the insecticidal metabolites were present, bioassays including resuspended spore-crystal pellets, filtered supernatants (FS) were compared with those of WC. Both fractions tested showed some level of insecticidal activity. The results may suggest that the main toxic factors can be found in FS and could be directly correlated with the presence of β-exotoxin. Based on the bioassays with FS and autoclaved FS, the participation of thermolabile virulence factors such as Cry1I in toxicity is neither discarded. In the selected strainsβ-exotoxin would be the major associated virulence factor; therefore, their use in biological control of A. grandis should be restricted. Nevertheless, these strains could be the source of genes (e.g., crylla) to produce transgenic cotton plants resistant to this pest.

Se realizaron ensayos preliminares con cultivos completos de 124 cepas de Bacillus thuringiensis utilizando larvas neonatas de Anthonomus grandis, una plaga principal del algodón en Argentina y otras regiones de América. Se seleccionaron 3 cepas exóticas y 4 nativas por producir mortalidad superior al 50%, todas ellas productoras de β-exotoxina. Las cepas nativas presentan la misma morfología de cristales, un perfil de proteínas similar y los mismos genes insecticidas. Estas características hacen que se parezcan a cepas tóxicas para lepidópteros. Además, mostraron un perfil de Rep-PCR idéntico al de la cepa lepidoptericida HD-1, lo que indica que podrían pertenecer al serovar kurstaki. Sin embargo, se observaron diferencias en el perfil plasmídico y en la producción de β-exotoxina. Para determinar en qué fracción del cultivo se encontraban los metabolitos responsables de la toxicidad, se compararon los resultados de bioensayos en los que se utilizó biomasa, sobrenadante filtrado (SF) o cultivos completos. Ambas fracciones mostraron cierto grado de toxicidad. Los resultados indican que los principales factores tóxicos se encuentran en el sobrenadante y estarían directamente relacionados con la presencia de β-exotoxina. De acuerdo con los bioensayos de SF y SF autoclavado, no se descarta también la participación en la toxicidad de factores de virulencia termolábiles, como CrylIa. En las cepas seleccionadas, el principal factor de virulencia es la β-exotoxina, por lo que su uso debería restringirse para el control biológico de A. grandis. No obstante, estas podrían ser fuente de genes (p. ej., crylIa) para la producción de plantas de algodón transgénicas resistentes a dicha plaga.

Asunto(s)

RESUMEN

Preliminary bioassays with whole cultures (WC) of 124 Bacillus thuringiensis strains were performed with neonate larvae of Anthonomus grandis, a major cotton pest in Argentina and other regions of the Americas. Three exotic and four native strains were selected for causing more than 50% mortality. All of them were ß-exotoxin producers. The native strains shared similar morphology of parasporal crystals, similar protein pattern and identical insecticidal gene profiles. These features resembled Lepidoptera-toxic strains. Furthermore, these strains showed a Rep-PCR pattern identical to lepidoptericidal strain HD-1, suggesting that these strains may belong to serovar kurstaki. However, some differences were observed in the plasmid profiles and in the production of ß-exotoxin. To determine the culture fractions where the insecticidal metabolites were present, bioassays including resuspended spore-crystal pellets, filtered supernatants (FS) were compared with those of WC. Both fractions tested showed some level of insecticidal activity. The results may suggest that the main toxic factors can be found in FS and could be directly correlated with the presence of ß-exotoxin. Based on the bioassays with FS and autoclaved FS, the participation of thermolabile virulence factors such as Cry1I in toxicity is neither discarded. In the selected strains, ß-exotoxin would be the major associated virulence factor; therefore, their use in biological control of A. grandis should be restricted. Nevertheless, these strains could be the source of genes (e.g., cry1Ia) to produce transgenic cotton plants resistant to this pest.

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