Controle biológico da lagarta do cartucho, Spodoptera frugiperda, com baculovírus.

2009

Levantamento dos danos causados pela infestação de Spodoptera frugiperda (J.E. Smith) em lavouras comerciais de milho Bt na Região Central de Minas Gerais.

2009

Monitoramento de parasitoides de lagartas de Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae) em municípios de Minas Gerais, Brasil.

2009

Aspectos morfológicos y biológicos de la oruga militar tardía (Spodoptera frugiperda (J.E. Smith) (Lep.: Noctuidae)

p.193-199

Desarrollo de un bioinsecticida a partir de Bacillus thuringiensis y extracto de Agave lechuguilla para el control de Spodoptera frugiperda (J. B. Smith)

Tesis (Doctor en Ciencias con Especialidad en

Constitutive oligomerization of human D-2 dopamine receptors expressed in Spodoptera frugiperda 9 (Sf9) and in HEK293 cells - Analysis using co-immunoprecipitation and time-resolved fluorescence resonance energy transfer

Human D-2Long (D-2L) and D-2Short (D-2S) dopamine receptor isoforms were modified at their N-terminus by the addition of a human immunodeficiency virus (HIV) or a FLAG epitope tag. The receptors were then expressed in Spodoptera frugiperda 9 (Sf9) cells using the baculovirus system, and their oligomerization was investigated by means of co-immunoprecipitation and time-resolved fluorescence resonance energy transfer (FRET). [H-3] Spiperone labelled D-2 receptors in membranes prepared from Sf9 cells expressing epitope-tagged D-2L or D-2S receptors, with a pK(d) value of approximate to 10. Co-immunoprecipitation using antibodies specific for the tags showed constitutive homo-oligomerization of D-2L and D-2S receptors in Sf9 cells. When the FLAG-tagged D-2S and HIV-tagged D-2L receptors were co-expressed, co-immunoprecipitation showed that the two isoforms can also form hetero-oligomers in Sf9 cells. Time-resolved FRET with europium and XL665-labelled antibodies was applied to whole Sf9 cells and to membranes from Sf9 cells expressing epitope-tagged D-2 receptors. In both cases, constitutive homo-oligomers were revealed for D-2L and D-2S isoforms. Time-resolved FRET also revealed constitutive homo-oligomers in HEK293 cells expressing FLAG-tagged D-2S receptors. The D-2 receptor ligands dopamine, R-(-) propylnorapomorphine, and raclopride did not affect oligomerization of D-2L and D-2S in Sf9 and HEK293 cells. Human D-2 dopamine receptors can therefore form constitutive oligomers in Sf9 cells and in HEK293 cells that can be detected by different approaches, and D-2 oligomerization in these cells is not regulated by ligands.

Analysis of the genome of Spodoptera frugiperda nucleopolyhedrovirus (SfMNPV-19) and of the high genomic heterogeneity in group II nucleopolyhedroviruses

The genome of the most virulent among 22 Brazilian geographical isolates of Spodoptera frugiperda nucleopolyhedrovirus, isolate 19 (SfMNPV-1 9), was completely sequenced and shown to comprise 132 565 bp and 141 open reading frames (ORFs). A total of 11 ORFs with no homology to genes in the GenBank database were found. Of those, four had typical baculovirus; promoter motifs and polyadenylation sites. Computer-simulated restriction enzyme cleavage patterns of SfMNPV-1 9 were compared with published physical maps of other SfMNPV isolates. Differences were observed in terms of the restriction profiles and genome size. Comparison of SfMNPV-1 9 with the sequence of the SfMNPV isolate 3AP2 indicated that they differed due to a 1427 bp deletion, as well as by a series of smaller deletions and point mutations. The majority of genes of SfMNPV-1 9 were conserved in the closely related Spodoptera exigua NPV (SeMNPV) and Agrotis segetum NPV (AgseMNPV-A), but a few regions experienced major changes and rearrangements. Synthenic maps for the genomes of group 11 NPVs revealed that gene collinearity was observed only within certain clusters. Analysis of the dynamics of gene gain and loss along the phylogenetic tree of the NPVs showed that group 11 had only five defining genes and supported the hypothesis that these viruses form ten highly divergent ancient lineages. Crucially, more than 60% of the gene gain events followed a power-law relation to genetic distance among baculoviruses, indicative of temporal organization in the gene accretion process.

Characterization of a beta-1,3-glucanase active in the alkaline midgut of Spodoptera frugiperda larvae and its relation to beta-glucan-binding proteins

Spodoptera frugiperda beta-1,3-glucanase (SLam) was purified from larval midgut. It has a molecular mass of 37.5 kDa, an alkaline optimum pH of 9.0, is active against beta-1,3-glucan (laminarin), but cannot hydrolyze yeast beta-1,3-1,6-glucan or other polysaccharides. The enzyme is an endoglucanase with low processivity (0.4), and is not inhibited by high concentrations of substrate. In contrast to other digestive beta-1,3-glucanases from insects, SLam is unable to lyse Saccharomyces cerevisae cells. The cDNA encoding SLam was cloned and sequenced, showing that the protein belongs to glycosyl hydrolase family 16 as other insect glucanases and glucan-binding proteins. Multiple sequence alignment of beta-1,3-glucanases and beta-glucan-binding protein supports the assumption that the beta-1,3-glucanase gene duplicated in the ancestor of mollusks and arthropods. One copy originated the derived beta-1,3-glucanases by the loss of an extended N-terminal region and the beta-glucan-binding proteins by the loss of the catalytic residues. SLam homology modeling suggests that E228 may affect the ionization of the catalytic residues, thus displacing the enzyme pH optimum. SLam antiserum reacts with a single protein in the insect midgut. Immunocytolocalization shows that the enzyme is present in secretory vesicles and glycocalyx from columnar cells. (C) 2010 Elsevier Ltd. All rights reserved.

The catalytic and other residues essential for the activity of the midgut trehalase from Spodoptera frugiperda

Trehalase (EC 3.2.1.28) hydrolyzes only alpha, alpha`- trehalose and is present in a variety of organisms, but is most important in insects and fungi. Crystallographic data showed that bacterial trehalase has 0312 and E496 as the catalytical residues and three Arg residues in the active site. Those residues have homologous in all family 37 trehalases including Spodoptera frugiperda trehalase (0322, E520, R169, R227, R287). To test the role of these residues, mutants of trehalase were produced. All mutants were at least four orders of magnitude less active than wild type trehalase and no structural difference between these mutants and wild type enzyme were discernible by circular dichroism. D322A and E520 pH-activity profile lacked the alkaline arm and the acid arm, respectively, suggesting that D322 is the acid and E520 the basic catalyst. Azide increases E520A activity three times, confirming its action as the basic catalyst. Taking into account the decrease in activity after substitution for alanine residue, the three arginine residues are as important as the catalytical ones to trehalase activity. This clarifies the previous misidentification of an Arg residue as the acid catalyst. As far as we know, this is the first report on the functional identification residues important for trehalase activity. (C) 2010 Elsevier Ltd. All rights reserved.

Sequencing of Spodoptera frugiperda midgut trehalases and demonstration of secretion of soluble trehalase by midgut columnar cells

Both soluble (SfTre1) and membrane-bound (SfTre2) trehalases occur along the midgut of Spodoptera frugiperda larvae. Released SfTre2 was purified as a 67 kDa protein. Its K(m) (1.6 mM) and thermal stability (half life 10 min at 62 degrees C) are different from the previously isolated soluble trehalase (K(m) = 0.47 mM; 100% stable at 62 degrees C). Two cDNAs coding for S. frugiperda trehalases have been cloned using primers based on consensus sequences of trehalases and having as templates a cDNA library prepared from total polyA-containing RNA extracted from midguts. One cDNA codes for a trehalase that has a predicted transmembrane sequence and was defined as SfTre2. The other, after being cloned and expressed, results in a recombinant trehalase with a K(m) value and thermal stability like those of native soluble trehalase. This enzyme was defined as SfTre1 and, after it was used to generate antibodies, it was immunolocalized at the secretory vesicles and at the glycocalyx of columnar cells. Escherichia coli trehalase 3D structure and sequence alignment with SfTre1 support a proposal regarding the residue modulating the pKa value of the proton donor.

Campoletis flavicincta (Hym., Ichneumonidae) : ocorrência, criação e interação com Spodoptera frugiperda (Lep., Noctuidae) e Bacillus thuringiensis aizawai

Spodoptera frugiperda é um inseto-praga responsável por altos níveis de desfolhamento em gramíneas cultivadas, sendo que, dentre os métodos de controle, o biológico pode vir a tornar-se uma alternativa. Foi feita uma revisão de literatura sobre parasitóides de S. frugiperda. A ocorrência de parasitóides desse inseto, em áreas de cultivo de milho da EEA/IRGA, foi avaliada em Cachoeirinha, RS. Verificou-se a presença de Chelonus sp., Cotesia sp. e Exaticolus sp. (Hym., Braconidae), Campoletis flavicincta e Ophion sp. (Hym., Ichneumonidae) e de Archytas incertus e Lespesia archippivora (Dip., Tachinidae), com predomínio de C. flavicincta. Em função da dificuldade de obtenção de fêmeas deste inseto em laboratório, foram avaliadas diferentes condições de criação. Desta forma, registrou-se uma razão sexual de 0,41 quando foram expostas lagartas de segundo ínstar de S. frugiperda, as fêmeas do parasitóide apresentavam idade entre 3 e 6 dias e os casais foram formados no momento da exposição. Por fim, aspectos referentes à interação entre C. flavicincta/S. frugiperda/B. thuringiensis aizawai, em laboratório, foram avaliados A partir de análise do consumo alimentar de folhas de milho, observou-se que lagartas parasitadas e infectadas apresentaram um menor consumo, apesar do mesmo não ter diferido daquele de lagartas apenas parasitadas. A mortalidade das lagartas parasitadas e infectadas foi superior tanto das infectadas quanto das parasitadas. Lagartas infectadas mostraram um período de alimentação que não diferiu das sadias, apesar de terem apresentado maior duração da fase larval. Indivíduos descendentes de casais que emergiram de lagartas infectadas não tiveram alteradas suas características biológicas. A análise histológica de lagartas parasitadas e infectadas indicou não ter havido alteração no ovo e larva do parasitóide, resultante da ação do bacilo. Pode-se, portanto, inferir que o uso conjunto do parasitóide e da bactéria não resulta em prejuízo para o parasitóide.

Biologia de Spodoptera frugiperda (J. E. Smith) (Lepidoptera: noctuidae) em genótipos de milho doce e comum

Spodoptera frugiperda, uma das principais pragas do milho, tem grande importância pelos danos e dificuldades de controle. A ausência de informações sobre seu impacto em milho doce motivou a comparação de sua biologia entre os genótipos ELISA, BR 400 (milho doce) e BR PAMPA (milho comum). Parcelas com estes genótipos foram estabelecidas em área experimental do Departamento de Fitossanidade, UFRGS, Porto Alegre, RS, de outubro-novembro/2000 e de outubro-novembro/2001. Em condições de laboratório (25±1ºC; 70±10%UR; fotofase 12 horas), lagartas foram individualizadas e alimentadas com seções de 3,14 cm2 de milho, totalizando 60 indivíduos/genótipo. Diariamente, avaliou-se a área foliar consumida, peso das lagartas, indivíduos mortos e recolheram-se as cápsulas cefálicas. As pupas foram sexadas e pesadas, computando-se os indivíduos mortos. Os adultos foram mantidos aos casais, registrando-se períodos de pré-oviposição, oviposição e pós-reprodutivo, número de posturas e de ovos/postura, período de incubação, intervalo entre oviposições e longevidade. A preferência alimentar foi avaliada quantificando-se o consumo foliar em observações de 12, 36 e 72 horas. Foram calculados índices nutricionais. Os dados foram submetidos à análise de variância e as médias comparadas por Tukey a 5%. Não se registrou diferença na largura das cápsulas, nos três primeiros ínstares. No quarto e quinto ínstares, as cápsulas das lagartas mantidas em BR 400 foram menores O consumo foliar, o peso das lagartas e das pupas, a duração dos ínstares e a razão sexual não diferiram entre os genótipos. A duração da fase pupal foi menor nas fêmeas mantidas em BR 400. A mortalidade foi maior, na fase larval, em ELISA e na pupal, em BR PAMPA. Na fase reprodutiva constatou-se diferença apenas no período de incubação, que em BR 400 foi mais longo. Não se observou preferência alimentar. Em relação aos índices nutricionais, somente a taxa de crescimento relativo (RGR) diferiu entre os genótipos, sendo que em BR 400 e ELISA foram observados valores superiores.

Biological activity of astilbin from Dimorphandra mollis against Anticarsia gemmatalis and Spodoptera frugiperda

Astilbin was isolated in high yield from Dimorphandra mollis, and its insecticidal and growth inhibiting activity by stomach ingestion were evaluated against Anticarsia gemmatalis and Spodoptera frugiperda. The insecticidal activity of astilbin, the weight reduction of the larval phase and the prolongation of the larval and pupal phases were verified for both species. Astilbin was identified on the base of its NMR, MS and physical data. (C) 2002 Society of Chemical Industry.

Associação de bioensaios e caracterização molecular para seleção de novos isolados de Bacillus thuringiensis efetivos contra Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)