Abstract Chrysodeixis includens has become the major Lepidopteran pest of soybean crops, especially in the Brazilian Cerrado (savanna) region. A native isolate of Chrysodeixis includens nucleopolyhedrovirus (ChinNPV) from this region, Buritis, MG, was assessed for its biological and molecular features. In addition, in vitro co-infection with Anticarsia gemmatalis multiple nucleopolyhedrovirus (AgMNPV), another virus of an important soybean pest, was tested. The ChinNPV-Buritis isolate presented an average LC50 of 7,750 occlusion bodies (OBs)/ml of diet in C. includens larvae. Analysis of restriction endonuclease profiles of viral DNA revealed similarities with previously described ChinNPV isolates IE, IF, and IG from Brazil, although the presence of submolar bands indicates genetic heterogeneity. Optical microscopy analysis in conjunction with quantitative PCR (qPCR) demonstrated in vitro infection of this isolate in IPLB-SF-21AE, Sf9, and BTI-Tn-5B1-4 cell lines, but the amount of ChinNPV tends to decrease through serial passages. The qPCR method developed in this study successfully detected both AgMNPV and ChinNPV from cell culture and from infected larvae. The cell line Tn-5B1-4 is indicated for future development of in vitro production and co-infection studies.

Resumo: O objetivo deste trabalho foi avaliar uma gama de hospedeiros, in vitro, quanto à resposta a Spodoptera frugiperda multiple nucleopolyhedrovirus (SfMNPV), um vírus patogênico à lagarta-do-cartucho (Spodoptera frugiperda; Lepidoptera: Noctuidae), para o posterior desenvolvimento de um biopesticida baseado em sistema de cultura de células. As linhagens celulares de Bombyx mori (BM-5), Lymantria dispar (IPLB-LD-625Y), Trichoplusia ni (BTI-Tn-5B1-4), Anticarsia gemmatalis (UFL-AG-286) e S. frugiperda (IPLB-SF-21AE e Sf9) foram testadas quanto a sua suscetibilidade a um isolado brasileiro de SfMNPV altamente virulento. Os efeitos citopáticos induzidos pelo vírus, a produção de partículas virais e a síntese de polipeptídeos virais foram examinados e comparados. Ambas as células de S. frugiperda apresentaram hipertrofia dos núcleos celulares e produção de muitos poliedros. O SDS-Page de proteínas com radiomarcação mostrou que as proteínas celulares apresentaram inibição de síntese, enquanto uma intensa banda de cerca de 30 kDa, reconhecida como poliedrina, era sintetizada. As outras linhagens celulares não apresentaram produção de poliedros, apesar de algumas terem apresentado alterações morfológicas e inibição de síntese proteica em resposta à infecção viral. As linhagens celulares SF-21 e Sf9 são recomendadas para a posterior produção in vitro de SfMNPV.

Abstract: The objective of this work was to evaluate the response of an in vitro host range to Spodoptera frugiperda multiple nucleopolyhedrovirus (SfMNPV), a pathogenic virus to the fall armyworm (Spodoptera frugiperda, Lepidoptera: Noctuidae), for the further development of a biopesticide based on cell culture systems. The cell lines from Bombyx mori (BM-5), Lymantria dispar (IPLB-LD-625Y), Trichoplusia ni (BTI-Tn-5B1-4), Anticarsia gemmatalis (UFL-AG-286), and S. frugiperda (IPLB-SF-21AE and Sf9) were tested for their susceptibility to a highly-virulent Brazilian isolate of SfMNPV. The cytopathic effects induced by the virus, the production of viral particles, and the synthesis of viral polypeptides were examined and compared. Both S. frugiperda cell lines showed hypertrophy of cell nuclei and production of many polyhedra. The SDS-Page of radiolabed proteins showed that the cell protein synthesis was shutoff, while an intense band of about 30 kDa, recognized as polyhedrin, was synthesized. The other cell lines did not show polyhedra production, although some of them underwent morphological changes and protein synthesis shutdown in response to virus infection. The SF-21 and Sf9 cell lines are recommended for further in vitro production of SfMNPV.

Os baculovírus são vírus patogênicos a insetos, encontrados principalmente na ordem Lepidoptera. A família Baculoviridae é taxonomicamente dividida em dois gêneros: Nucleopolyhedrovirus e Granulovirus, que diferem pela morfologia do corpo de oclusão. Os nucleopoliedrovírus (NPV) possuem corpos de inclusão poliédrica (PIB), contendo múltiplas partículas virais, enquanto os granulovírus (GV) contêm, em geral, partículas únicas, ocluídas em corpos protéicos de forma ovóide. Durante o ciclo de vida são produzidos dois tipos de progênies virais: BV ("Budded Virus") e PDV ("Polyhedra Derived Virus"), que são essenciais para o processo de infecção e propagação do vírus. Os baculovírus têm sido empregados no controle de pragas e, por serem específicos e restritos a invertebrados, são considerados agentes seguros de controle biológico. Recentemente têm sido amplamente utilizados como vetor de expressão de genes heterólogos por produzirem e processarem, em grande quantidade, proteínas de procariotos e eucariotos. Além disso, técnicas de DNA recombinante têm permitido a produção de inseticidas virais geneticamente modificados. Este trabalho constitui uma revisão sobre a taxonomia, estrutura, replicação e biologia molecular de baculovírus e sobre seu uso como bioinseticida no Brasil.

Baculoviruses are insect viruses found mainly in Lepidoptera. The family Baculoviridae is taxonomically divided in two genera, Nucleopolyhedrovirus and Granulovirus, which differ by occlusion body morphology. NPVs (Nucleopolyhedroviruses) have polyhedrical inclusion bodies (PIBs) containing multiple viral particles, while GVs (Granuloviruses) appear to be generally single particles occluded in oval shaped occlusion bodies. During the life cycle, two different viral progenies are produced: BV (Budded Virus) and PDV (Polyhedra Derived Virus), which are essential for the infectious process and virus propagation in host cells. Baculoviruses are being used for pest control and they are especially safe due to their specificity and invertebrate-restricted host range. Baculoviruses have been used as vectors for high level protein expression ofheterologous genes from prokaryotic and eukaryotic organisms. Also, recombinant DNA techniques have allowed the production of genetically modified viral insecticides. This study is a review on the taxonomy, structure, replication and molecular biology of baculoviruses, as well as their use as bioinsecticides in Brazil.