Does the aggressiveness of the prey modify the attack behavior of the predator Supputius cincticeps (Stål) (Hemiptera, Pentatomidae)?

Does the aggressiveness of the prey modify the attack behavior of the predator Supputius cincticeps (Stål) (Hemiptera, Pentatomidae)? The stink bug Supputius cincticeps (Stål) (Hemiptera, Pentatomidae) is a predator found in several Brazilian regions, which possesses desirable attributes as a natural control agent and in biological control programs. The aim of this study was to test if the attack behavior and predation success of S. cincticeps were affected by prey species. Larvae of Tenebrio molitor (L.) (Coleoptera, Tenebrionidae), Spodoptera frugiperda (J. E. Smith) (Lepidoptera, Noctuidae), and Thyrinteina arnobia (Stoll) (Lepidoptera, Geometridae) were offered to S. cincticeps in laboratory bioassays where predatory attack and prey defensive behaviors were observed for 2-hour periods. The attack behavior of S. cincticeps changed with the prey species offered. More than 25% of T. molitor and S. frugiperda larvae were immediately attacked, but T. arnobia was not immediately attacked by S. cincticeps. Successful attack (i.e., successful insertion of the predator stylets into the prey) depends on the region of the body attacked, with a greater proportion of successful attacks in the anterior than in the median or posterior regions. Larvae of T. arnobia and S. frugiperda displayed a sequence of abrupt head and body movements in response to S. cincticeps attack. Attempts of predation were more successful on T. molitor and S. frugiperda than on T. arnobia. Information about the differential attack behavior of S. cincticeps on different prey species is important for designing successful biological control programs using this hemipteran predator.

Indirect effect of neem oil on Podisus nigrispinus (Hemiptera, Pentatomidae): biology and predatory capacity

This study evaluated the effects on the development and predatory capacity of Podisus nigrispinus fed on Spodoptera frugiperda that have ingested different concentrations of neem oil. The predatory capacity of Podisus nigrispinus was assessed, separating nymphs (fourth instar) and adults (males and females). The treatments consisted of S. frugiperda larvae reared in neem oil aqueous solutions (0.077, 0.359 and 0.599%), deltamethrin EC 25 (0.100%) and control arranged in a completely randomized design, with ten replicates. Insects were offered three larval densities (one, three and six), in the third or fourth instars. The predated larvae were examined at 24 and 48 hours after the beginning of the experiment. Biological parameters of Podisus nigrispinus were evaluated in groups of ten second-instar nymphs transferred to pots, in five replicates. Insects were offered 2-6 third and/or fourth-instar larvae reared in the same neem oil concentrations in a completely randomized design. The following parameters were evaluated: duration of each nymph stage (days), nymph mortality (%), weight of fifth-instar nymphs (mg), sex ratio, weight of males and females (mg) and longevity of unfed adults (days). The predatory capacity of nymphs and adults of Podisus nigrispinus was influenced by the neem oil at the concentrations of 0.359% and 0.599% in the highest density. The concentration of 0.359% lengthened the nymphal stage and the concentration of 0.599% reduced the weight of males.

In vivo binding of the Cry11Bb toxin of Bacillus thuringiensis subsp. medellin to the midgut of mosquito larvae (Diptera: Culicidae)

Bacillus thuringiensis subsp. medellin produces numerous proteins among which 94 kDa known as Cry11Bb, has mosquitocidal activity. The mode of action of the Cry11 proteins has been described as similar to those of the Cry1 toxins, nevertheless, the mechanism of action is still not clear. In this study we investigated the in vivo binding of the Cry11Bb toxin to the midgut of the insect species Anopheles albimanus, Aedes aegypti, and Culex quinquefasciatus by immunohistochemical analysis. Spodoptera frugiperda was included as negative control. The Cry11Bb protein was detected on the apical microvilli of the midgut epithelial cells, mostly on the posterior midgut and gastric caeca of the three mosquito species. Additionally, the toxin was detected in the Malpighian tubules of An. albimanus, Ae. aegypti, Cx. quinquefasciatus, and in the basal membrane of the epithelial cells of Ae. aegypti midgut. No toxin accumulation was observed in the peritrophic membrane of any of the mosquito species studied. These results confirm that the primary site of action of the Cry11 toxins is the apical membrane of the midgut epithelial cells of mosquito larvae.

Novo isolado de Bacillus thuringiensis efetivo contra a lagarta-do-cartucho

Com o objetivo de encontrar princípios ativos que possam ser usados no controle da lagarta-do-cartucho, Spodoptera frugiperda (Smith, 1797), foram testados 205 isolados de Bacillus, provenientes de diferentes regiões brasileiras, contra larvas deste inseto. Apenas um isolado de B. thuringiensis subsp. kurstaki, denominado S93, causou 100% de mortalidade. A CL50 da mistura esporos-cristais do isolado S93 para larvas de 3º estádio de S. frugiperda foi de 37,0 ng/mL, enquanto a da estirpe HD-1 de B. thuringiensis subsp. kurstaki isolada do produto Dipel foi de 177,73 mg/mL, demonstrando ser o novo isolado muito mais tóxico contra a lagarta-do-cartucho do que HD-1-Dipel.

Relação entre a lagarta-do-cartucho e seus agentes de controle biológico natural na produção de milho

Este trabalho teve como objetivo quantificar as perdas na cultura do milho BRS 3123 decorrentes do ataque da lagarta-do-cartucho (Spodoptera frugiperda), na ausência e presença de seus inimigos naturais. Aos 15 dias depois da emergência da planta, foi realizada uma infestação artificial da praga (uma postura por metro quadrado). As lagartas alimentaram-se da planta, sem a interferência de seus inimigos naturais, pela utilização de uma gaiola de proteção (inicial, de dois dias depois da infestação, até uma proteção máxima de 16 dias). O dano da praga, determinado por uma escala de notas de 0 (plantas sem dano) a 5 (plantas mortas), foi em média de 4,01, 1,39, 1,09 e 0,93 para o período de proteção da praga de 16, 6, 4 e 2 dias, respectivamente. Na ausência de agentes de controle biológico, o ataque da praga ocasionou perdas na produção de matéria seca de 47,27% e perdas no rendimento de grãos de 54,49%. Os resultados evidenciaram a importância dos inimigos naturais na supressão de lagartas de S. frugiperda na cultura de milho.

Molecular characterization and pathogenicity of isolates of Beauveria spp. to fall armyworm

The objective of this work was to evaluate the pathogenicity of 24 Beauveria isolates to Spodoptera frugiperda larvae, and characterize them molecularly through rDNA-ITS sequencing and RAPD markers. Sequencing of rDNA-ITS fragments of 570 bp allowed the identification of isolates as B. bassiana or B. brongniarti by sequence comparison to GenBank. Sixty seven polymorphic RAPD fragments were capable to differentiate 20 among 24 Beauveria isolates, grouping them according to the derived host insect and to pathogenicity against maize fall armyworm larvae. Three RAPD markers were highly associated to the pathogenicity against S. frugiperda, explaining up to 67% of the phenotypic variation. Besides identification and molecular characterization of Beauveria isolates, ITS sequence and RAPD markers proved to be very useful in selecting the isolates potentially effective against S. frugiperda larvae and in monitoring field release of these microorganisms in biocontrol programs.

Respostas da lagarta‑do‑cartucho a milho geneticamente modificado expressando a toxina Cry 1A(b)

O objetivo deste trabalho foi avaliar parâmetros biológicos da lagarta-do-cartucho do milho (Spodoptera frugiperda), alimentada com híbridos de milho Bt, que expressam a toxina Cry 1A(b), e com seus respectivos isogênicos não Bt. Os experimentos foram realizados no laboratório da Embrapa Milho e Sorgo, em Sete Lagoas, MG. Os parâmetros avaliados foram: sobrevivência de larvas após 48 horas, sobrevivência da fase larval e pré-imaginal, biomassa de larvas aos 14 dias de idade, biomassa de pupas, período de desenvolvimento larval, e não preferência alimentar de larvas do primeiro ínstar. Larvas de S. frugiperda apresentam menor sobrevivência nas primeiras 48 horas de alimentação e durante toda a fase larval, na maioria dos híbridos de milho Bt, em comparação ao milho não Bt. A biomassa de larvas e pupas foi sempre menor no milho Bt, e o período larval e o pré-imaginal, maior. Houve interação entre a toxina Cry 1A(b) e a base genética dos híbridos transgênicos, quanto à sobrevivência e à biomassa larval. Larvas recém-eclodidas de S. frugiperda apresentam preferência pela alimentação em milho não Bt.

Avaliação de tecnologias de refúgio no cultivo de milho transgênico

O objetivo deste trabalho foi comparar o uso de refúgio tradicional em áreas separadas com a tecnologia alternativa de refúgio pela mistura de sementes não transgênicas no saco ("refuge in the bag" - RIB) em diferentes proporções. Foram utilizados os híbridos comerciais transgênicos AG 7000YG e DKB 390YG com refúgios plantados com as respectivas cultivares convencionais. Avaliaram-se sete tratamentos: RIB com quatro proporções de mistura de sementes não transgênicas (2,5, 5,0, 7,5 e 10%) no saco; refúgio tradicional, com 10% da área plantada exclusivamente com cultivar não transgênica; e área totalmente cultivada com plantas transgênicas ou totalmente com plantas não transgênicas. O híbrido DKB 390YG foi o mais produtivo. Para este híbrido, não foram observadas diferenças de produtividade entre os tratamentos, com exceção do controle inteiramente convencional, que produziu menos. Com o híbrido AG 7000YG, a tecnologia RIB com proporções de 5,0 e 7,5% de sementes não transgênicas apresentou as maiores produtividades, significativamente superiores às do refúgio tradicional, que não diferiu do controle convencional. Nos tratamentos RIB, as diferenças nas intensidades de dano por Spodoptera frugiperda não se refletiram em diferenças na produtividade. O refúgio no saco é alternativa viável para substituir o método de refúgio utilizado atualmente.

Caracterização do gene vip3A e toxicidade da proteína Vip3Aa50 à lagarta-do-cartucho e à lagarta-da-soja

O objetivo deste trabalho foi caracterizar o gene vip3A de Bacillus thuringiensis e verificar a toxicidade da proteína Vip3Aa50 a larvas da lagarta-do-cartucho (Spodoptera frugiperda) e da lagarta-da-soja (Anticarsia gemmatalis). O gene vip3A foi amplificado por PCR, com iniciadores específicos, e gerou um fragmento de 2.370 pb. Esse fragmento foi clonado em vetor pGEM-T Easy e, em seguida, sequenciado, subclonado em vetor de expressão pET-28a (+) e inserido em células de Escherichia coli BL21 (DE3). A expressão da proteína Vip3Aa50 foi induzida por isopropil-β-D-1-tiogalactopiranosídeo (IPTG), visualizada em SDS-PAGE e detectada por "Western blot". Os ensaios de toxicidade revelaram alta atividade da proteína Vip3Aa50 contra as larvas neonatas da lagarta-da-soja e da lagarta-do-cartucho, com CL50 de 20,3 e 79,6 ng cm-2, respectivamente. O gene vip3Aa50 é um novo gene da classe vip3A.

Manejo de lepidópteros-praga na cultura do milho com o evento Bt piramidado Cry1A.105 e Cry2Ab2

O objetivo deste trabalho foi avaliar a eficácia do evento piramidado (MON 89034), que expressa as proteínas Cry1A.105 e Cry2Ab2, no controle dos principais lepidópteros-praga da cultura do milho no Brasil, Spodoptera frugiperda, Helicoverpa spp. e Diatraea saccharalis. Os ensaios foram conduzidos em quatro regiões do país, com o híbrido DKB 390, submetido a seis tratamentos: híbrido com o evento piramidado, híbrido com o evento que expressa apenas a proteína Cry1A(b) (MON 810) e híbrido convencional (não Bt), todos com e sem manejo integrado de S. frugiperda. Para o evento piramidado, não foi necessário o controle químico em nenhum dos locais avaliados. Diferenças significativas foram observadas entre os tratamentos quanto aos danos e à presença de lagartas. Em geral, essas variáveis foram mais baixas no híbrido com o evento piramidado e mais altas no híbrido convencional, sem controle químico. Sob alta infestação, o controle químico reduziu os danos causados por S. frugiperda e D. saccharalis, tanto no evento que expressa apenas uma proteína, como no híbrido convencional. Com base nos danos causados pelos insetos, o evento piramidado Cry1A.105 e Cry2Ab2 é eficiente no controle dos principais lepidópteros-pragas do milho no Brasil.

Relação entre toxicidade de proteínas Vip3Aa e sua capacidade de ligação a receptores intestinais de lepidópteros-praga

Resumo:O objetivo deste trabalho foi avaliar a toxicidade de novas proteínas Vip3Aa e sua capacidade de ligação a vesículas de membrana da microvilosidade apical (VMMA) do intestino de lagartas neonatas de Spodoptera frugiperda, Anticarsia gemmatalise Heliothis virescens. Proteínas expressas pelos genes vip3Aa42 e vip3Aa43 mostraram-se tóxicas a S. frugiperda (CL50 de 78,2 e 113 ng cm-2, respectivamente) e A. gemmatalis(CL50 de 239,2 e 57,5 ng cm-2, respectivamente), e pouco tóxicas a H. virescens (CL50>5.000 ng cm-2). Os ensaios de ligação às VMMA mostraram que as proteínas unem-se de forma efetiva aos receptores nas vesículas das espécies avaliadas, mas essa capacidade de ligação somente é efetiva na ativação da toxicidade para as populações avaliadas de S. frugiperdae A. gemmatalis.

Constituintes químicos e atividade inseticida dos extratos de frutos de Trichilia elegans E T. catigua (Meliaceae)

Phytochemical investigation of the fruits extracts of Trichilia elegans and Trichilia catigua (Meliaceae) has led to the identification of the limonoids 11β-acetoxyobacunone, cedrelone, methylangolensate and epimeric mixture of photogedunin besides known coumarins (scoparone, scopoletin, umbeliferone) and the steroids stigmasterol, β-sitosterol, sitostenone and campesterol. The structures of the compounds were proposed by spectroscopic analysis and comparison with literature data. An evaluation of the insecticidal activity of the fruits extracts of Trichilia ssp. was carried out and the extracts of T. elegans revealed to have strong insecticidal activity and the extracts of T. catigua showed moderate larval mortality on Spodoptera frugiperda.

Corn yield response to weed and fall armyworm controls

The interference imposed the by weeds on corn decreases practically all vegetative characteristics. As consequence, the green ear and grain yield are also reduced. Losses due to the fall armyworm (Spodoptera frugiperda) attack can reduce corn grain yield up to 34%. In general, weed and insect control issues are addressed separately in research papers. Nevertheless, interaction between weeds and insects may exist. This study aimed to evaluate green ear and corn grain yield response to weed and fall armyworm control. A completely randomized block design with split-plots and five replicates was adopted. Corn cultivar AG 1051 was grown under weedy conditions or with control by hand hoeings performed at 20 and 40 days after planting. Fall armyworm control (applied to subplots) was performed with sprays of water (control), deltamethrin (5g active ingredient ha-1); neem oil, at 0.5% (diluted in water), and neem leaf extract at 5%. Each product was sprayed three times, at seven-day intervals, starting at the 7th day after planting, using 150 L ha-1 of the tank solution. Dry mass of the above-ground part, internode diameter, leaf length, leaf width, leaf area, green ear yield and grain yield of corn were reduced due to the lack of weed control. Fall armyworm control in the weeded plots did not influence green ear yield and grain yield, except green mass of marketable, husked ears, which was reduced when the caterpillar was not controlled. Without weed control, neem extracts and deltamethrin sprays provided highest yields of number and total weight of green ears with husks, number and weight of marketable ears with husks and number of marketable ears without husks. The best results for husked ear mass and for grain yield were obtained with neem extract and deltamethrin, respectively.

Characterization of the interdependency between residues that bind the substrate in a β-glycosidase

The manner by which effects of simultaneous mutations combine to change enzymatic activity is not easily predictable because these effects are not always additive in a linear manner. Hence, the characterization of the effects of simultaneous mutations of amino acid residues that bind the substrate can make a significant contribution to the understanding of the substrate specificity of enzymes. In the β-glycosidase from Spodoptera frugiperda (Sfβgly), both residues Q39 and E451 interact with the substrate and this is essential for defining substrate specificity. Double mutants of Sfβgly (A451E39, S451E39 and S451N39) were prepared by site-directed mutagenesis, expressed in bacteria and purified using affinity chromatography. These enzymes were characterized using p-nitrophenyl β-galactoside and p-nitrophenyl β-fucoside as substrates. The k cat/Km ratio for single and double mutants of Sfβgly containing site-directed mutations at positions Q39 and E451 was used to demonstrate that the effect on the free energy of ES‡ (enzyme-transition state complex) of the double mutations (∆∆G‡xy) is not the sum of the effects resulting from the single mutations (∆∆G‡x and ∆∆G‡y). This difference in ∆∆G‡ indicates that the effects of the single mutations partially overlap. Hence, this common effect counts only once in ∆∆G‡xy. Crystallographic data on β-glycosidases reveal the presence of a bidentate hydrogen bond involving residues Q39 and E451 and the same hydroxyl group of the substrate. Therefore, both thermodynamic and crystallographic data suggest that residues Q39 and E451 exert a mutual influence on their respective interactions with the substrate.

Interaction between Telenomus remus and Trichogramma pretiosum in the management of Spodoptera spp.

Interaction betweeen Telenomus remus and Trichogramma pretiosum in the management of Spodoptera spp. The use of egg parasitoids is a promising strategy for Integrated Pest Management (IPM), but different species of parasitoids have greater or lesser control efficiency, depending on the pest species. Recently, not only Anticarsia gemmatalis and Pseudoplusia includens but also Spodoptera cosmioides and S. eridania have been among the key Lepidoptera larvae attacking soybeans. This study evaluated the combination of Telenomus remus and Trichogramma pretiosum for parasitism of eggs of the Spodoptera complex, for better control efficiency and broader spectrum of action among the key pests of soybeans. The experiment was carried out under controlled environmental conditions (25 ± 2ºC; 70 ± 10% RH; and 14 h photophase) in a completely randomized experimental design with seven treatments and 10 replicates with S. frugiperda, S. cosmioides and S. eridania eggs. Each replicate consisted of one egg mass of each Spodoptera species, with approximately 100 eggs offered to the parasitoids. The treatments were: 1) 10 females of T. pretiosum; 2) nine females of T. pretiosum and one female of T. remus; 3) eight females of T. pretiosum and two females of T. remus; 4) seven females of T. pretiosum and three females of T. remus; 5) six females of T. pretiosum and four females of T. remus; 6) five females of T. pretiosum and five females of T. remus, and 7) 10 females of T. remus. The parameter evaluated was the percentage of parasitized eggs. Results showed that treatments combining both parasitoid species with only 1 T. remus for each 9 T. pretiosum (10%) and only 2 T. remus for each 8 T. pretiosum (20%) were enough to significantly increase the parasitism observed on eggs of S. cosmioides and S. frugiperda, respectively. This association of T. pretiosum and T. remus in different proportions is very promising for biological control in IPM programs because it provides wide spectrum of control.