Phylogeography of Chelonus insularis (Hymenoptera: Braconidae) and Campoletis sonorensis (Hymenoptera: Ichneumonidae), Two Primary Neotropical Parasitoids of the Fall Armyworm (Lepidoptera: Noctuidae)

In a previous study, we observed no spatial genetic structure in Mexican populations of the parasitoids Chelonus insularis Cresson (Hymenoptera: Braconidae) and Campoletis sonorensis Cameron (Hymenoptera: Ichneumonidae) by using microsatellite markers In the current study, we Investigated whether for these important parasitoids of the fall armyworm (Lepidoptera: Noctuidae) there is any genetic structure at a larger scale Insects of both species were collected across the American continent and their phylogeography was Investigated using both nuclear and mitochondria] markers Our results suggest an ancient north-south migration of C insularis, whereas no clear pattern] could be determined for C sonorensis. Nonetheless, the resulting topology indicated the existence of a cryptic taxon within this later species. a few Canadian specimens determined as C. sonorensis branch outside a clack composed of the Argentinean Chelonus grioti Blanchard, the Brazilian Chelonus flavicincta Ashmead, and the rest of the C sonorensis individuals The individuals revealing the cryptic taxon were collected from Thichoplusia in (Hubner) (Lepidoptera. Noctuidae) on tomato (Lycopersicon spp) and may represent a biotype that has adapted to the early season phenology of its host. Overall, the loosely defined spatial genetic structure previously shown at a local fine scale also was found at the larger scale, for both species Dispersal of these insects may be partly driven by wind as suggested by genetic similarities between Individuals coming from very distant locations.

Feeding non-preference by Spodoptera frugiperda and Spodoptera eridania on tomato genotypes

Larvae of the genus Spodoptera spp. are highly polyphagous and can cause economical losses in several agricultural crops. Given their growing importance in the tomato crop, especially for industry, this work aimed to evaluate the feeding non-preference by larvae of Spodoptera frugiperda (J. E. Smith, 1797) and Spodoptera eridania (Cramer, 1782) on tomato genotypes and classify them by the levels of resistance. The commercial cultivar Santa Clara was set as the susceptible standard and line PI 134417 as the resistant standard to evaluate the lines PI 134418, PI 126931, LA 462 and LA 716. Feeding non-preference tests were performed under non-choice and free-choice conditions to evaluate the genotype attractiveness to larvae at predetermined times after their release, as well as the leaf area consumed. Overall, the genotypes LA 716 and PI 126931 were the least attractive to S. frugiperda, whereas Santa Clara was the most attractive and consumed. For S. eridania, the genotypes PI 126931, LA 462, LA 716 and PI 134418 were the least preferred for feeding, and Santa Clara and PI 134417 were the most attractive and consumed. The genotypes LA 716 and PI 126931 are moderately resistant to S. frugiperda and S. eridania; PI 134418 and LA 462 are moderately resistant to S. eridania; PI 134417 is susceptible to S. frugiperda and S. eridania; and Santa Clara is highly susceptible to both S. frugiperda and S. eridania.

Combate à lagarta do cartucho Spodoptera frugiperda (J.E.Smith, 1797), em plantação de milho

Com a finalidade de estudar o eleito de alguns defensivos químicos sobre a "lagarta do cartucho do milho" foi instalado o presente ensaio no "Campo de Produção de Mudas Tietê", da Coordenadoria de Assistência Técnica Integral (CATI), em Tietê, SP. Foram utilizados os seguintes tratamentos: A - testemunha; B- bendiocarbe, 250 g C- hidrazona BTS 54439, 250 g; D - triclorfom, 250 g; E - piretróide bifentrina (FMC 54800), 30 g; F- piretróide (FMC - 65318), 30 g; G - tiodicarbe, 200 g; H - triazofós, 200 g. As quantidades são de ingrediente ativo por hectare. Os tratamentos tiveram quatro repetições, em blocos ao acaso, com parcelas de 150 m² (cada tratamento, 600 m² ). Aos dois e nove dias da aplicação, os piretróides (FMC - 54800 (bifentrina) e FMC 65318) e tiodicarbe foram promissores.

Eficiência de alguns produtos químicos sobre a "lagarta do cartucho do milho" Spodoptera frugiperda (J.E.Smith, 1797)

A lagarta do cartucho do milho é um sério depredador da cultura do milho. Quase todos os anos, os danos ocasionados às plantas são grandes. Para se conhecer o efeito de alguns defensivos sobre a citada lagarta, realizou-se este trabalho experimental, instalado em Piracicaba, SP, a 09 de dezembro de l983. Os produtos foram aplicados com pulverizador costal, manual, de 20 litros de capacidade, provido de bico em leque Teejet 8002. Utilizaram-se 200 litros de água por hectare e 30 cm³ de "Extravon" (espalhante-adesivo) por 100 litros de calda. Os tratamentos foram nove: A - testemu nha; B - monocrotofós, 240 g; C - bifentrina, 10 g; D - triazofós, 200 g; E -carbamato UC 75118, 150 g; F - monocro tofós, 400 g; G - acefato, 375 g; H - cabamato UC 76724, 150 g; I - clofentezina, 250 g. As quantidades acima são de ingrediente ativo por hectare. Aos 02 dias da pulverização, os melhores foram o triazofós e a bifentrina. Aos 07 dias, nenhum produto apresentava boa eficiência.

CONTROLE INTEGRADO DE SPODOPTERA FRUGIPERDA (SMITH & ABBOTT) UTILIZANDO-SE O PARASITÓIDE TELENOMUS REMUS NIXON

Este estudo foi conduzido em 1996 e 1997 para avaliar a eficiência da liberação de Telenomus remus Nixon (cerca de 200.000 adultos/ha), sozinho ou integrado ao vírus de poliedrose nuclear de Spodoptera frugiperda (VPNSf) ou a um inseticida químico seletivo, para o controle de Spodoptera frugiperda (Smith & Abbott). Os experimentos foram conduzidos em delineamento experimental de blocos ao acaso, com seis tratamentos e quatro repetições. Foram avaliados os danos provocados pela praga às folhas (escala de 0 a 5) e o rendimento de espigas. De maneira geral, os danos provocados pela praga foram significativamente superiores nas parcelas testemunhas (nota média de 2,94). Não houve diferença significativa entre os demais tratamentos (média 1,04). De maneira semelhante, houve diferença significativa no rendimento de espiga entre testemunha (7.165 kg/ha) e demais tratamentos (9.084 kg/ha). Não houve efeito dos tratamentos no comprimento da espiga e no dano às espigas.

Atividade de extrato aquoso de folhas de nim (Azadirachta indica) sobre Spodoptera frugiperda

A lagarta-do-cartucho do milho (Spodoptera frugiperda (J. E. Smith)) tem sido controlada com inseticidas sintéticos. Uma das caracteristicas do nim (Azadirachta indica A. Juss) é sua atividade inseticida contra pragas, como sucedâneo aos sintéticos. O objetivo deste trabalho foi avaliar a atividade inseticida do extrato aquoso das folhas do nim sobre a lagarta-do-cartucho do milho, em laboratório. Bioensaios com diferentes concentrações de extrato em dieta artificial, tendo o inseticida chlorpyrifos como testemunha, revelaram, 15 dias após infestação com larvas, eficiência equivalente entre as concen- trações 3,60 a 10,00 mg mL-1. A análise de Probit mostrou CL50 = 2,67 mg mL-1; o extrato aquoso das folhas de nim apresenta, portanto, efeito inseticida sobre a lagarta-do-cartucho do milho.

Diversidade molecular entre populações de Spodoptera frugiperda no Brasil avaliada por marcadores AFLP

O objetivo deste trabalho foi avaliar a diversidade molecular, por meio de marcadores AFLP, de seis populações de Spodoptera frugiperda coletadas na cultura do milho, em diferentes regiões geográficas do Brasil. O DNA foi extraído de lagartas de quarto instar, e as reações de AFLP foram realizadas com sete combinações de oligonucleotídeos iniciadores. A partir das seis populações de S. frugiperda estudadas, foi identificado um grupo principal formado por três populações geneticamente mais relacionadas. As populações de S. frugiperda analisadas mostram alta variabilidade genética, com máximo de 58% de similaridade.

Atividade inseticida dos frutos de Trichilia claussenii (Meliaceae) sobre Spodoptera frugiperda

An evaluation of the insecticidal activity of the fruits extracts of Trichilia claussenii was carried out and the methanol extract revealed to have strong insecticidal activity. The fractionation of methanol extract of T. claussenii seeds bioassay-guided against Spodoptera frugiperda has led to the identification of the ω-phenylalkyl and alkenyl fatty acids as active compounds in this extract. The structures of the compounds were proposed by spectroscopic analysis and comparison with literature data.

Atividade inseticida de óleos essenciais de Pelargonium graveolens l'Herit e Lippia alba (Mill) N. E. Brown sobre Spodoptera frugiperda (J. E. Smith)

Insecticidal activity of essential oils of Pelargonium graveolens, Lippia alba and compounds geraniol, linalool, 1,8-cineole, limonene, carvone, citral and Azamax® were evaluated against Spodoptera frugiperda. Topical application assay showed essential oil of P. graveolens has acute toxicity against Spodoptera frugiperda larvae (third instar) with LD50 1.13 µg/mg per insect and LD90 2.56 µg/mg per insect. Three essential oils of L. alba also exhibited insecticidal activity with LD50 ranging from 1.20 to 1.56 µg/mg per insect and LD90 from 2.60 to 3.75 µg/mg per insect. Geraniol, linalool, carvone and citral caused significant mortality of 30, 90, 84 and 64% respectively, compared to negative control. The bioinsecticide, Azamax®, caused lower mortality than the compounds of the essential oils.

Actividad insecticida sobre Spodoptera frugiperda (Lepidóptera: Noctuidae) de los compuestos aislados de la parte aérea de Piper septuplinervium (Miq.) c. dc. y las inflorescencias de Piper subtomentosum Trel. & Yunck. (Piperaceae)

The bioassay-guided purification of ethanolic extracts from inflorescences of Piper subtomentosum Trel. & Yunck and aerial part of Piper septuplinervium (Miq. ) C. DC. led to isolation of five flavonoids, uvangoletin (1), galangin (2), chrysin (5), 5-hydroxy-4',7-dimethoxy-flavone (6), pinostrobin (7); one amide, N-p-coumaroil-tyramine (4); one acylglycerol, monopalmitin (3); one derivative of acid, protocatechuic acid (8); and glycosydated sterol, daucosterol (9). Their structures were elucidated on the basis of spectroscopy and spectrometry data and by comparison with data reported in the literature. The isolated compounds were tested against Spodoptera frugiperda. The results showed galangin and protocatechuic acid to be the most active (LC 50 13.63 and 17.16 ppm, respectively).

Volumes de calda e pontas de pulverização no controle químico de Spodoptera frugiperda na cultura do sorgo forrageiro

No manejo de Spodoptera frugiperda na cultura do sorgo, a utilização de inseticida ainda é a principal tática recomendada, contudo este método de controle precisa ser otimizado. O presente trabalho teve como objetivo avaliar o efeito da utilização de três pontas e dois volumes de calda na aplicação de inseticida para o controle químico de Spodoptera frugiperda na cultura do sorgo. O ensaio foi conduzido no delineamento em blocos ao acaso, com quatro repetições, em esquema fatorial (3 x 2) + 1: três tipos de pontas de pulverização (jato cônico vazio, jato plano defletor duplo e jato plano defletor com indução de ar), dois volumes de calda (200 e 130 L ha-1) e um tratamento adicional que não recebeu inseticida. Realizou-se a semeadura direta de uma cultivar de sorgo forrageiro, avaliando-se, após a aplicação do inseticida clorpirifós, a deposição de calda no dossel da cultura, o controle de Spodoptera frugiperda e a produtividade. Pôde-se concluir que as pontas de jato plano defletor duplo e jato plano defletor com indução de ar proporcionaram maior cobertura do dossel das plantas de sorgo do que a ponta de jato cônico, resultando em maior eficácia de controle de Spodoptera frugiperda e maior produtividade. O volume de calda de 200 L ha-1; quando comparado ao volume de 130 L ha-1; também proporcionou melhor eficácia de controle.

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

Antifeedant activities of crude seed extracts of tropical African spices against Spodoptera littoralis (Lepidoptera: Noctuidae)

The antifeedant activities of Piper guineense Schum et Thonn (Piperaceae), Aframomum melegueta (Rosk) K. Schum (Zingiberaceae), Aframomum citratum (Pareira) K. Schum (Zingiberaceae) and Afrostyrax kamerunensis Perkins and Gilg (Huaceae) seed extracts were investigated in laboratory dual- and no-choice bioassays using third-instar Spodoptera littoralis (Boisduval) larvae. In the dual-choice test, the hexane and methanol extracts of A. melegueta showed potent dose-dependent antifeedant activity at concentrations of ≥300 ppm and the water extract at ≥500 ppm, as illustrated by significantly lower leaf consumptions. Aframomum citratum methanol and water extracts exhibited antifeedant activity at ≥300 and ≥1000 ppm, respectively, but the hexane and ethanol extracts did not affect feeding at any concentration. Piper guineense ethanol and water extracts showed dose-dependent antifeedant effects at ≥300 and ≥500 ppm, respectively, and the methanol extract was active only at 1000 ppm. None of the extracts of the highly aromatic A. kamerunensis exhibited antifeedant activity at any of the tested concentrations. In the no-choice bioassays, extracts with antifeedant activity in the dual-choice tests also showed dose-dependent feeding inhibition. The hexane and methanol extracts of A. melegueta were effective in the no-choice tests at ≥100 and ≥500 ppm, respectively, and the water extract at ≥300 ppm. Similarly, the A. citratum water and methanol extracts were active at ≥500 ppm and the P. guineense water and ethanol extracts at ≥100 ppm. GC/MS chromatography of A. melegueta hexane and methanol extracts revealed volatile constituents with known anti-insect activity. The hexane and methanol extracts of A. melegueta, the methanol extract of A. citratum and the water and ethanol extracts of P. guineense may have potential for use by subsistence farmers.

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.