A betabaculovirus encoding a gp64 homolog.

BACKGROUND: A betabaculovirus (DisaGV) was isolated from Diatraea saccharalis (Lepidoptera: Crambidae), one of the most important insect pests of the sugarcane and other monocot cultures in Brazil. RESULTS: The complete genome sequence of DisaGV was determined using the 454-pyrosequencing method. The genome was 98,392 bp long, which makes it the smallest lepidopteran-infecting baculovirus sequenced to date. It had a G + C content of 29.7% encoding 125 putative open reading frames (ORF). All the 37 baculovirus core genes and a set of 19 betabaculovirus-specific genes were found. A group of 13 putative genes was not found in any other baculovirus genome sequenced so far. A phylogenetic analysis indicated that DisaGV is a member of Betabaculovirus genus and that it is a sister group to a cluster formed by ChocGV, ErelGV, PiraGV isolates, ClanGV, CaLGV, CpGV, CrleGV, AdorGV, PhopGV and EpapGV. Surprisingly, we found in the DisaGV genome a G protein-coupled receptor related to lepidopteran and other insect virus genes and a gp64 homolog, which is likely a product of horizontal gene transfer from Group 1 alphabaculoviruses. CONCLUSION: DisaGV represents a distinct lineage of the genus Betabaculovirus. It is closely related to the CpGV-related group and presents the smallest genome in size so far. Remarkably, we found a homolog of gp64, which was reported solely in group 1 alphabaculovirus genomes so far.

High susceptibility and low resistance allele frequency of Chrysodeixis includens (Lepidoptera: Noctuidae) field populations to Cry1Ac in Brazil.

BACKGROUND: The soybean looper (SBL), Chrysodeixis includens (Walker), is one of the most important soybean pests in Brazil. MON 87701 × MON 89788 soybean expressing Cry1Ac has been recently deployed in Brazil, providing high levels of control against the primary lepidopteran pests. To support insect resistance management (IRM) programmes, the baseline susceptibility of SBL to Cry1Ac was assessed, and the resistance allele frequency was estimated on the basis of an F2 screen. RESULTS: The toxicity (LC50 ) of Cry1Ac ranged from 0.39 to 2.01 µg mL(-1) diet among all SBL field populations collected from crop seasons 2008/09 to 2012/13, which indicated approximately fivefold variation. Cry1Ac diagnostic concentrations of 5.6 and 18 µg mL(-1) diet were established for monitoring purposes, and no shift in mortality was observed. A total of 626 F2 family lines derived from SBL collected from locations across Brazil during crop season 2014/15 were screened for the presence of Cry1Ac resistance alleles. None of the 626 families survived on MON 87701 × MON 89788 soybean leaf tissue (joint frequency 0.0004). CONCLUSIONS: SBL showed high susceptibility and low resistance allele frequency to Cry1Ac across the main soybean-producing regions in Brazil. These findings meet important criteria for effective IRM strategy. © 2015 Society of Chemical Industry.

High genetic stability of peroral infection factors from Anticarsia gemmatalis MNPV over 20years of sampling.

The Anticarsia gemmatalis multiple nucleopolyhedrovirus (AgMNPV) has been used as a biopesticide since the early 1980s in Brazil to control the major pest of soybean crops, the velvetbean caterpillar, Anticarsia gemmatalis. To monitor the genetic diversity over space and time we sequenced four pif genes (pif1, pif2, pif3 and pif4) from AgMNPV isolates collected from different regions of South America, as well as of seasonal isolates, sampled during a two-decade field experiment. Although all genes presented low levels of polymorphism, the pif-2 carries a slightly higher number of polymorphic sites. Overall, this study reveals that pif genes have remained stable after 20 years of repeated field application.

Histology and ultrastructure of the fat body of Anticarsia gemmatalis (HÜBNER, 1818) (LEPIDOPTERA: NOCTUIDAE)

The aim of this study was to analyze the morphologically the fatty body of fourth-instar Anticarsia gemmatalis larvae under light, transmission and scanning electron microscopy. Two distinct portions of the fat body were detected: the parietal (PA) and perivisceral (PV). The PA, the parietal portion, presented a long-stripped shape located below the tegument and lateral to the digestive tube. The PV, rarely observed, was in dorsal region, adhered to digestive wall. Both the portions were constituted of only one cellular type, the trophocytes. These cells in the PA were organized in one layer of thickness showing cylindrical contiguously morphology, whereas the PV was comprised by a mass of small cells, superposed as clusters. Both the portions were covered by a layer of connective tissue, grouping the trophocytes and keeping them separated from the hemolymph. The cytoplasm of the trophocytes from the PA presented acidophilic stain, while the basophilic cytoplasmic of the trophocytes from the PV was due to the large amount of rough endoplasmic reticulum. From the results, it could be concluded that the fat body presented morphological and ultrastructural differences according to the portion and that these features could characterize distinct functions.

Soybean response to different injury levels at early developmental stages / Resposta da cultura da soja a diferentes intensidades de injúria nos estágios iniciais de desenvolvimento

This study evaluated the effects of different soybean injury levels at early growth stages of the plants. The experiments were carried out in two different growing seasons, using a randomized complete block design, with 11 treatments in the first and 10 treatments in the second season, and four replications. The 'BRS 133' soybean cultivar was used, and the injury levels tested were: 1-Control; 2-Removal of 1 cotyledon; 3-Removal of both cotyledons; 4-Removal of both cotyledons + 1 unifoliate leaf; 5-Removal of both cotyledons + both unifoliate leaves; 6-Removal of 1 unifoliate leaf; 7-removal of both unifoliate leaves; 8-Cut below unifoliate leaves; 9-Removal of trifoliate leaf sprouts; 10-Total defoliation; 11-Total defoliation + removal of apical bud (only in the first trial). Injury was manually imposed, and insecticides were applied weekly to prevent injury by insects. The results showed that the soybean yield was reduced only when the injury was severe enough to cause plant stand reductions as a consequence of plant death. This occurred for treatments 5, 10 and 11. Under the other treatments, the soybean plants were able to recover. These findings show that the recommended economic threshold of 30% defoliation to initiate pest control is safe, and should be accepted by growers.

Este estudo avaliou os efeitos de diferentes níveis de injúria nos estádios iniciais de desenvolvimento das plantas de soja. Os experimentos foram realizados em duas diferentes safras agrícolas em delineamento de blocos completos casualizados, com 11 tratamentos na primeira e 10 tratamentos na segunda safra, e quatro repetições. A cultivar utilizada foi a 'BRS 133' e os níveis de injúria foram: 1-Testemunha; 2-Remoção de 1 cotilédone; 3-Remoção de ambos os cotilédones; 4-Remoção de ambos os cotilédones + 1 folha unifoliolada; 5-Remoção de ambos cotilédones + ambas as folhas unifolioladas; 6-Remoção de 1 folha unifoliolada; 7-Remoção de ambas as folhas unifolioladas; 8-Corte abaixo das folhas unifoliadas; 9-Remoção do broto das folhas trifolioladas; 10-Desfolha total; 11-Desfolha total + remoção da gema apical (apenas no primeiro ensaio). A injúria foi realizada manualmente e inseticidas foram aplicados semanalmente para impedir injúria por insetos. Os resultados evidenciaram que a produção de soja foi reduzida somente quando a injúria foi severa o suficiente para provocar a redução do estande em consequência da morte de plantas. Isso ocorreu nos tratamentos 5, 10 e 11. Nos demais tratamentos, as plantas foram capazes de se recuperar, o que comprova que o nível de ação de 30% de desfolha que é recomendado para iniciar o controle é seguro e deve ser respeitado pelos sojicultores.

The role of peritrophic membrane in the resistance of Anticarsia gemmatalis larvae (Lepidoptera: Noctuidae) during the infection by its nucleopolyhedrovirus (AgMNPV).

The aim of this study was to analyze morphologically the peritrophic membrane (PM) of Anticarsia gemmatalis larvae resistant (RL) and non-resistant (susceptible) (SL) to the A. gemmatalis multicapsid nucleopolyhedrovirus (AgMNPV), in the presence of viral infection. Also, in this investigation the results between SL and RL were compared to improve the understanding of the resistance mechanisms to the virus. The PM of SL of A. gemmatalis was less efficient as a barrier against the viral infection since it was found to be more fragile than the PM of RL. The lower chitin content as seen from weaker fluorescent staining in SL as well as the abundance of non-solubilized vesicular materials in the ectoperitrophic space, would cause the malformation of this membrane, facilitating the passage of the virus toward the epithelium of the midgut. On the other hand, in RL, the intensity of WGA (wheat germ agglutinin)-conjugated FITC (fluorescein) reaction of the PM was greater than in SL, making this insect more resistant to infection. We can conclude that the effectiveness of the PM in protecting against pathogens is dependent on the integrity of the epithelial cells of the midgut and of the structural preservation of the PM, being directly implicated in the resistance of A. gemmatalis larvae to AgMNPV.

Lepidopteran larva consumption of soybean foliage: basis for developing multiple-species economic thresholds for pest management decisions.

BACKGROUND: Defoliation by Anticarsia gemmatalis (Hübner), Pseudoplusia includens (Walker), Spodoptera eridania (Cramer), S. cosmioides (Walker) and S. frugiperda (JE Smith) (Lepidoptera: Noctuidae) was evaluated in four soybean genotypes. A multiple-species economic threshold (ET), based upon the species' feeding capacity, is proposed with the aim of improving growers' management decisions on when to initiate control measures for the species complex. RESULTS: Consumption by A. gemmatalis, S. cosmioides or S. eridania on different genotypes was similar. The highest consumption of P. includens was 92.7 cm(2) on Codetec 219RR; that of S. frugiperda was 118 cm(2) on Codetec 219RR and 115.1 cm(2) on MSoy 8787RR. The insect injury equivalent for S. cosmoides, calculated on the basis of insect consumption, was double the standard consumption by A. gemmatalis, and statistically different from the other species tested, which were similar to each other. CONCLUSIONS: As S. cosmioides always defoliated nearly twice the leaf area of the other species, the injury equivalent would be 2 for this lepidopteran species and 1 for the other species. The recommended multiple-species ET to trigger the beginning of insect control would then be 20 insect equivalents per linear metre.

Genetic diversity of the sunflower caterpillar (Chlosyne lacinia saundersii Doubleday and Hewitson) (Lepidoptera: Nymphalidae) populations determined by molecular RAPD markers.

Chlosyne lacinia saundersii is one of the most important pests of sunflower and it is the main target of insecticides applications. Larvae were collected in Londrina (PR), Santa Maria (RS), Dourados (MS), Ribeirão Preto (SP), Brasília (DF), Barreiras (BA), Uberaba (MG) and Vilhena (RO). Genomic DNA was extracted and amplified with ten-mer primers, which produced 101 loci. The size of the RAPD amplicons ranged from 180 to 2564 bp. Polymorphism among populations ranged from 31% to 67%, with the highest polymorphisms of 57% and 67% being detected in Uberaba and Vilhena populations, respectively. Populations with the highest similarity determined with Dice coefficient were from Ribeirão Preto and Barreiras, while insects from Londrina showed the highest similarity among them. Gene flow of C. lacinia saundersii 1.1 was lower than those previously observed for the noctuid Anticarsia gemmatalis Hübner, suggesting that C. lacinia saundersii populations are more isolated than the ones of this noctuid. Through the Analysis of Molecular Variance (AMOVA), RAPD variance was 33.64% among geographical populations and 66.36% within populations. These results suggest that populations of C. lacinia saundersii are genetically structured.

Genetic diversity of the sunflower caterpillar (Chlosyne lacinia saundersii Doubleday and Hewitson) (Lepidoptera: Nymphalidae) populations determined by molecular RAPD markers

Chlosyne lacinia saundersii is one of the most important pests of sunflower and it is the main target of insecticides applications. Larvae were collected in Londrina (PR), Santa Maria (RS), Dourados (MS), Ribeirão Preto (SP), Brasília (DF), Barreiras (BA), Uberaba (MG) and Vilhena (RO). Genomic DNA was extracted and amplified with ten-mer primers, which produced 101 loci. The size of the RAPD amplicons ranged from 180 to 2564 bp. Polymorphism among populations ranged from 31 percent to 67 percent, with the highest polymorphisms of 57 percent and 67 percent being detected in Uberaba and Vilhena populations, respectively. Populations with the highest similarity determined with Dice coefficient were from Ribeirão Preto and Barreiras, while insects from Londrina showed the highest similarity among them. Gene flow of C. lacinia saundersii 1.1 was lower than those previously observed for the noctuid Anticarsia gemmatalis Hübner, suggesting that C. lacinia saundersii populations are more isolated than the ones of this noctuid. Through the Analysis of Molecular Variance (AMOVA), RAPD variance was 33.64 percent among geographical populations and 66.36 percent within populations. These results suggest that populations of C. lacinia saundersii are genetically structured.

Chlosyne lacinia saundersii é uma das mais importantes pragas da cultura do girassol e o principal alvo das aplicações de inseticidas. As larvas foram coletadas em Londrina (PR), Santa Maria (RS), Dourados (MS), Ribeirão Preto (SP), Brasília (DF), Barreiras (BA), Uberaba (MG) e Vilhena (RO). O DNA genômico foi extraído e amplificado com dez primers, que produziram 101 locos. O tamanho das amplificações de RAPD variou de 180 a 2564 pb. O polimorfismo entre as populações variou de 31 por cento a 67 por cento, com maior polimorfismo 57 por cento e 67 por cento, detectado em populações de Uberaba e Vilhena, respectivamente. As populações com maior similaridade determinada com o coeficiente de Dice foram de Ribeirão Preto e Barreiras, enquanto os insetos coletados em Londrina apresentaram maior similaridade entre eles. O fluxo gênico de C. lacinia saundersii de 1,1 foi menor que o observado para a Anticarsia gemmatalis Hübner Noctuidae, sugerindo que as populações de C. lacinia saundersii estão mais isoladas do que estes noctuideos. Através da análise de variância molecular (AMOVA), RAPD a variação foi de 33,64 por cento entre as populações geográficas e 66,36 por cento dentro das populações. Estes resultados sugere que as populações de C. lacinia saundersii são geneticamente estruturadas.

Evaluation of seven viral isolates as potential biocontrol agents against Pseudoplusia includens (Lepidoptera: Noctuidae) caterpillars.

The caterpillar Pseudoplusia includens (Walker, 1857) (Lepidoptera, Noctuidae), known as soybean looper, is a pest that has recently assumed greater importance in soybean in Brazil. Isolates of nucleopolyhedroviruses (NPVs) of this pest have been identified from cotton in Guatemala and soybean farms in Brazil, providing an interesting perspective of potential use of viral insecticide against the insect in lieu to chemical insecticides. With the objective to contribute to the characterization studies of this virus, morphological and molecular analyses and biological activity were carried out with seven P. includens viral isolates (I-A to I-G). Electron microscopy of viral samples, purified from macerated infected larvae, showed particles with typical morphology of the Baculoviridae family, genus Alphabaculovirus (Nucleopolyhedrovirus - NPV) presenting virions with only a single nucleocapsid per envelope (SNPV) occluded in a protein matrix, forming occlusion bodies (OB). This virus was then classified as P. includens single nucleopolyhedrovirus (PsinSNPV). OB particles analyzed in SDS-polyacrylamide gel showed an intense band corresponding in size to NPV polyhedrin protein. DNA restriction profiles of the PsinSNPV isolates showed differences in the fragment size and number suggesting the existence of genotypic variants, except between I-E and I-F profiles that were similar. Among the isolates tested for infectivity against P. includens, I-A, I-E and I-F were the most virulent. Survival times (ST(50)) varied according to viral concentration, with significant differences among isolates for the three higher concentrations.

Hemocyte quantitative changes in Anticarsia gemmatalis (Lepidoptera: Noctuidae) larvae infected by AgMNPV

The initial effects of the infection by AgMNPV in the total and differential counts of the hemocytes in Anticarsia gemmatalis (Lepidoptera: Noctuidae) larvae were studied. The total number of the hemocytes did not decrease in infected larvae, as it occurred in non infected larvae. In infected larvae, the hemocyte types showed the following frequencies: plasmatocytes - 47.8 percent, esferulocytes - 25.9 percent, granulocytes - 15.8 percent, oenocytoids - 7.2 percent, prohemocytes - 2.8 percent, vermicytes - 0,5 percent. Only the percentage of the granulocytes was different among infected and non infected larvae, indicating that these cells responded quickly to the initial viral infection. These results showed the effective role of the hemocytes in the response of the A. gemmatalis to the infection by AgMNPV. The comprehension of the immunological mechanisms of this insect is an important tool to understand its biological control.

Os efeitos iniciais da infecção por AgMNPV nas contagens total e diferencial dos hemócitos em Anticarsia gemmatalis (Lepidoptera: Noctuidae) foram estudados. O número total de hemócitos não diminuiu nas larvas infectadas, como ocorreu nas larvas não infectadas. Nas larvas infectadas, os tipos de hemócitos apresentaram as seguintes freqüências: plasmatócitos - 47,8 por cento, esferulócitos - 25,9 por cento, granulócitos - 15,8 por cento, oenocitóides - 7,2 por cento, prohemócitos - 2,8 por cento, vermiformes - 0,5 por cento. Apenas a porcentagem de granulócitos foi diferente entre larvas infectadas e não infectadas, indicando que estas células responderam rapidamente à infecção viral inicial. Estes resultados mostraram o papel efetivo que dos hemócitos na resposta de A. gemmatalis à infecção por AgMNPV. A compreensão dos mecanismos imunológicos deste inseto é uma ferramenta importante para compreender seu controle biológico.

Interactions among insect-resistant soybean genotypes extracts with populations of Anticarsia gemmatalis Hübner (Lepidoptera: Noctuidae) susceptible and resistant to its nucleopolyhedrovirus

Anticarsia gemmatalis nucleopolyhedrovirus (AgMNPV) is being used in Brazil as a biological insecticide. Host plant resistance of soybean to insects is been searched for and some authors have mentioned the interference of plant chemistry in virus efficiency. Interactions among soybean extracts of genotypes used as a source of resistance (PI 274454 and PI 227687) with different AgMNPV concentrations in populations of A. geatalis susceptible (S) and resistant (R) to the virus were studied at laboratory condition. Higher mortality was observed when larvae fed on diets with extracts of the soybean genotypes compared with those fed on a plain diet (control). The mean lethal concentration (LC50) was reduced about 10 ties in the S-population fed on diets containing PI 274454 extracts and different concentrations of AgMNPV, compared to control diet. Additive effect was predominantly observed when larvae fed on diets with extracts of soybean genotypes (PI 274454 and PI 227687) and AgMNPV for both larval populations. The pupal weight was negatively influenced by the extracts incorporated to the diets compared to control, for both larval populations, notably for R-population. The results suggest that, in general, leaf extracts of soybean resistant genotype did not cause any harmful effect on virus efficiency.

O nucleopoliedrovirus de Anticarsia gemmatalis (AgMNPV) tem sido utilizado como um inseticida biológico no Brasil. A resistência de plantas de soja a insetos tem sido pesquisada e alguns autores têm mencionado a interferência de substâncias químicas de plantas sobre a eficiência de vírus. As interações entre extratos de genótipos de soja utilizados como fontes de resistência (PI 274454 e PI 227687) com diferentes concentrações do AgMNPV em populações de A. gemmatalis suscetível (S) e resistente (R) ao vírus foram estudadas em condições de laboratório. Mortalidades elevadas foram observadas quando as larvas foram alimentadas com dietas contendo extratos dos genótipos de soja, em relação às larvas alimentadas com dieta artificial sem a presença de extratos (testemunha). A concentração letal média (CL50) foi reduzida em aproximadamente 10 vezes, na população s alimentada com dieta contendo extratos da PI 274454 e diferentes concentrações do AgMNPV, comparada à dieta testemunha. Um efeito aditivo foi predominantemente observado quando as larvas se alimentaram em dietas com extratos dos genótipos de soja (PI 274454 e PI 227687) e o AgMNPV, para ambas as populações (S e R). O peso de pupa foi negativamente influenciado pela dieta contendo os extratos em relação à dieta testemunha, para ambas as populações, com destaque para a população R. Os resultados indicam que, no geral, os extratos de folhas de genótipos de soja resistentes não causam efeitos negativos na eficiência do vírus.

Interactions among insect-resistant soybean genotypes extracts with populations of Anticarsia gemmatalis Hübner (Lepidoptera: Noctuidae) susceptible and resistant to its nucleopolyhedrovirus.

Anticarsia gemmatalis nucleopolyhedrovirus (AgMNPV) is being used in Brazil as a biological insecticide. Host plant resistance of soybean to insects is been searched for and some authors have mentioned the interference of plant chemistry in virus efficiency. Interactions among soybean extracts of genotypes used as a source of resistance (PI 274454 and PI 227687) with different AgMNPV concentrations in populations of A. geatalis susceptible (S) and resistant (R) to the virus were studied at laboratory condition. Higher mortality was observed when larvae fed on diets with extracts of the soybean genotypes compared with those fed on a plain diet (control). The mean lethal concentration (LC50) was reduced about 10 ties in the S-population fed on diets containing PI 274454 extracts and different concentrations of AgMNPV, compared to control diet. Additive effect was predominantly observed when larvae fed on diets with extracts of soybean genotypes (PI 274454 and PI 227687) and AgMNPV for both larval populations. The pupal weight was negatively influenced by the extracts incorporated to the diets compared to control, for both larval populations, notably for R-population. The results suggest that, in general, leaf extracts of soybean resistant genotype did not cause any harmful effect on virus efficiency.

Identification of a new nucleopolyhedrovirus from naturally-infected Condylorrhiza vestigialis (Guenée) (Lepidoptera: Crambidae) larvae on poplar plantations in South Brazil.

A baculovirus was isolated from larvae of Condylorrhiza vestigialis (Guenée) (Lepidoptera: Crambidae), a pest of a forest species known as Poplar (family Salicaceae, genus: Populus) with high economic value. Electron microscopy analysis of the occlusion body obtained from diseased larvae showed polyhedra containing multiple nucleocapsids per envelope. This baculovirus was thus named Condylorrhiza vestigialis multiple nucleopolyhedrovirus (CoveMNPV) and characterized by its DNA restriction endonuclease pattern, polyhedral protein, viral protein synthesis, and infectivity in insect cell lines. Restriction endonuclease profiles of viral DNA digested with five restriction enzymes were obtained and the CoveMNPV genome size was estimated to be 81+/-2.5 kbp. The isolation of the polyhedra (OBs) was done from the crude extract of infected larvae by ultracentrifugation through sucrose gradients. These viral particles were analyzed by denaturing polyacrylamide gel electrophoresis (SDS-PAGE), which showed a strong band with approximately 33 kDa, corresponding to the main protein of the occlusion bodies (polyhedrin). Also, a similar band was observed for CoveMNPV infected Spodoptera frugiperda cells (SF-21 AE) pulse-labeled with [(35)S] methionine and fractionated by SDS-PAGE. Of the four insect cell lines tested for susceptibility to CoveMNPV infection, the SF-21 AE was the most susceptible with occlusion bodies produced in most of the inoculated cells. This is the first record of an NPV from C. vestigialis.

A morphometric study of the midgut in resistant and non-resistant Anticarsia gemmatalis (Hübner) (Lepidoptera: Noctuidae) larvae to its nucleopolyhedrovirus (AgMNPV).

In this investigation, the anterior and posterior regions of the midgut of resistant (RL) and non-resistant (SL) Anticarsia gemmatalis larvae were analyzed morphometrically to characterize different regions along their length. Also, this investigation compares the results between SL and RL to improve the understanding of the resistance mechanisms to the virus. Histological sections were analyzed in a computerized system and the data were statistically analyzed by the Kruskal-Wallis test and by multivariate analysis. The midguts are morphometrically different in the two larval populations; we observed higher values in RL. The morphometric analysis of the epithelial cells showed that only columnar and goblet cells were distinct along the midgut, in both larvae, with the higher values found in the anterior region. Comparing the results between the two larval populations, all the epithelial cells presented significant differences, with RL showing the higher morphometric values. We concluded that there are regional differences along the length of midgut in SL and RL that confirm the idea of two morpho-functional distinct regions. The consistently morphometric superior values in RL indicate that this variability can be related with the resistance of A. gemmatalis to its AgMNPV.

Susceptibility/resistance of Anticarsia gemmatalis larvae to its nucleopolyhedrovirus (AgMNPV): Structural study of the peritrophic membrane.

This investigation compares the peritrophic membrane (PM) morphology along the midgut of susceptible (SL) and resistant (RL) Anticarsia gemmatalis larvae to the AgMNPV. The PM increased the thickness from the anterior to the posterior midgut region in both insects strain; however, the intensity of FITC-WGA reaction of the PM in the RL were greater than in SL. The PM in RL was ultrastructurally constituted by several layers of fibrous/vesicular materials in comparison with the few ones in SL. Our results showed that the structure of PM in the RL could be one of the resistance barriers to AgMNPV.

Genome of the most widely used viral biopesticide: Anticarsia gemmatalis multiple nucleopolyhedrovirus.

The genome of Anticarsia gemmatalis multiple nucleopolyhedrovirus isolate 2D (AgMNPV-2D), which is the most extensively used virus pesticide in the world, was completely sequenced and shown to have 132 239 bp (G+C content 44.5 mol%) and to be capable of encoding 152 non-overlapping open reading frames (ORFs). Three ORFs were unique to AgMNPV-2D, one of which (ag31) had similarity to eukaryotic poly(ADP-ribose) polymerases. The lack of chiA and v-cath may explain some of the success and growth of the AgMNPV biological control programme, as it may explain the high recovery of polyhedra sequestered inside dead larvae in the field, which are collected and used for further application as biological pesticides in soybean fields. The genome organization was similar to that of the Choristoneura fumiferana defective MNPV (CfDefNPV). Most of the variation between the two genomes took place near highly repetitive regions, which were also closely associated with bro-coding regions. The separation of the NPVs into groups I and II was supported by: (i) a phenogram of the complete genomes of 28 baculovirus and Heliothis zea virus 1, (ii) the most parsimonious reconstruction of gene content along the phenograms and (iii) comparisons of genomic features. Moreover, these data also reinforced the notion that group I of the NPVs can be split further into the AgMNPV lineage (AgMNPV, CfDefNPV, Epiphyas postvittana NPV, Orgyia pseudotsugata MNPV and C. fumiferana MNPV), sharing eight defining genes, and the Autographa californica MNPV (AcMNPV) lineage (AcMNPV, Rachiplusia ou NPV and Bombyx mori NPV), sharing nine defining genes.

Are chemical compounds important for soybean resistance to Anticarsia gemmatalis?

The identification and quantification of flavonoids (rutin and genistin) present in extracts of soybean genotypes, and their effects on the biology and physiology of Anticarsia gemmatalis Hübner (Lep.: Noctuidae) were studied. Analysis of covariance and bicoordinate utilization plots were used to remove the effect of feeding time from pupal weight and consumption as well as to separate pre- and postingestive effects of treatment on A. gemmatalis growth. Genotypes PI 274454, PI 227687, and "IAC-100" extracts in general, caused higher mortality, negatively influenced initial larval and pupal weight, and elongated larval cycle. Larvae fed on the "IAC-100" extract diet ingested larger amounts of food per unit of time, but were less efficient in its conversion to biomass. Leaf extracts of PI 227687 had the largest concentration of rutin (quercitin 3-O-rhamnosylglucoside), followed by PI 274454, and "IAC-100"; PI 74454 also had the highest genistin (genistein 7-O-glucoside) content. The susceptible cultivar "BR-16" showed only a kaempferol-based flavonoid in its chemical profile, indicating that after successive crosses, secondary compounds responsible for plant defenses were eliminated. Genotypes PI 274454, PI 227687, and "IAC-100" showed accentuated resistance characteristics and were considered inadequate sources for the development of A. gemmatalis. Considering rutin and genistin concentration in these genotypes, it is suggested that flavonoids are important factors conferring resistance to A. gemmatalis.

Produção de anticorpos policlonais para lectina de hemolinfa de Anticarsia gemmatalis / Production of polyclonal antibodies for lectin from Anticarsia gemmatalis hemolymph

A lagarta de Anticarsia gemmatalis promove extensos danos na cultura da soja e seu controle é geralmente baseado na aplicação de inseticidas químicos. Devido aos riscos à saúde humana, animal e ao meio ambiente, métodos alternativos de controle tem sido desenvolvidos como o bioinseticida Baculovirus anticarsia. Há relatos de desenvolvimento de resistência em populações de A. gemmatalis submetidas, em laboratório, ao tratamento com baculovirus durante várias gerações. Os insetos apresentam mecanismos elaborados de proteção contra agentes infecciosos, como as lectinas, que atuam como moléculas de reconhecimento. Assim, o objetivo deste estudo foi desenvolver anticorpos policlonais para lectina de A. gemmatalis. A atividade de lectina de hemolinfa de lagartas de A. gemmatalis foi avaliada frente a hemácias humanas, de coelho, camundongo, carneiro e boi de ensaio de hemaglutinação. Apenas as hemácias de bovino nåo foram aglutinadas pela lectina. As hemácias de coelho apresentaram maior reatividade com a lectina (1:512) e portanto os anticorpos policlonais foram produzidos em coelho imunizado com hemácias autólogas sensibilizadas com lectina. O anticorpo anti-lectina apresentou título de 1:8 em reação de precipitação em gel. Assim neste estudo foi possível produzir anticorpos para lectina de A. gemmatalis sem necessidade de emprego de técnicas dispendiosas de purificação.

Morphological study of the hindgut in larvae of Anticarsia gemmatalis Hübner(Lepidoptera: Noctuidae) / Estudo morfológico do intestino posterior em larvas de Anticarsia gemmatalis Hübner (Lepidoptera: Noctuidae)

The velvetbean caterpillar (Anticarsia gemmatalis Hübner) has great economical interest as it affects the soybean crop worldwide. This work describes the morphology of the hindgut in A. gemmatalis larvae emphasizing their histological aspects. Distinct morphological regions, identified as pylorus, ileum, colon and rectum, constitute the hindgut of A. gemmatalis. A thin cuticular intima, a simple epithelium and muscular layer compose the hindgut wall, independent of the region. Microspines project from the cuticular intima in the posterior interstitial ring, between the midgut and the hindgut, and the posterior pyloric region. A single circular layer of large fibers, differing from the other hindgut regions that present two layers of muscular fibers, forms the rectal musculature. The distal ends of Malpighian tubules cross the rectum wall and constitute the cryptonephric excretory system typical in Lepidoptera larvae.

A lagarta da soja (Anticarsia gemmatalis Hübner) tem grande interesse econômico, pois afeta significativamente a cultura da soja em todo o mundo. Este trabalho descreve a morfologia do intestino posterior de larvas de A. gemmatalis, com ênfase nos seus aspectos histológicos. O intestino posterior é constituído por regiões morfologicamente distintas, identificadas como piloro, íleo, cólon e reto. Independente da região, a parede do intestino posterior é constituída por fina cutícula, epitélio simples e camada muscular. A íntima cuticular apresenta espículas no anel intersticial posterior, entre o intestino médio e o posterior, e na região posterior do piloro. A musculatura do reto é formada por camada única de largas fibras circulares, diferindo das demais regiões do intestino posterior que apresentam duas camadas de fibras musculares. As extremidades distais dos túbulos de Malpighi atravessam as paredes do reto, constituindo o sistema criptonefridial característico de Lepidoptera.