Estudo da variabilidade entomopatogênicos nematóides populações (Heterorhabditidae) da Argentin

Entomopathogenic nematodes (EPN) belonging to the Heterorhabditidae family are lethal parasites of soil-dwelling insects. Two species were reported in Argentina: Heterorhabditis argentinensis and Heterorhabditis bacteriophora characterized mainly by morphometric features. In this work a comparative and phylogenetic study between five Heterorhabditis populations from Argentina was conducted to analyze the variability between strains and to evaluate the taxonomic position of Heterorhabditis argentinensis. The PCA analyses of morphometric characters separated the larger juvenile, female and male H. argentinensis from H. bacteriophora populations. The juvenile (IJs) stage provided the clearest separation of Heterorhabditis populations presenting the least variability between strains. The variable L and MBW were highly related to H. argentinensis IJs. Three groups were separated by this stage considering PC1 and PC2: one formed by H. bacteriophora OLI, RIV and RN strains, (isolates from Córdoba and Río Negro province), one for H. bacteriophora VELI strain (Buenos Aires province) and one for H. argentinensis (Santa Fe province). Heterorhabditis bacteriophora VELI and H. argentinensis isolated from regions with more rainfalls and humidity presented larger values for morphometric features. Molecular analyses showed the Argentinian populations (H. bacteriophora VELI strain and H. argentinensis), forming a same clade, with six other H. bacteriophora populations (not from Argentina) with a genetic similarity between them of 99%. Heterorhabditis argentinensis presented one unique nucleotide that was not present in any of the other species of the clade. Considering the results of this study H. argentinensis would be conspecific to H. bacteriophora, constituting a strain with a great morphometric variation where the host and climatic conditions could have influenced on the measurements.Nematóides entomopatogênicos (EPN) pertencentes à família Heterorhabditidae são parasitas letais de insetos que vivem no solo. Duas espécies foram relatados na Argentina: Heterorhabditis argentinensis e Heterorhabditis bacteriophora, caracterizada principalmente por características morfométricas. Neste trabalho um estudo comparativo e filogenética entre cinco populações do Heterorhabditis da Argentina foi conduzido para analisar a variabilidade entre as linhagens e avaliar a posição taxonômica das Heterorhabditis argentinensis. Características morfométricas de Heterorhabditis bacteriophora VELI e H. argentinensis isoladas de regiões com mais chuvas e umidade apresentaram dimensões maiores. Analisa o PCA de personagens morfométricas separou a maior juvenil, feminino e masculino H. argentinensis de H. bacteriophora populações. A fase juvenil (JIs) fornece a mais clara separação de populações Heterorhabditis apresentando a menor variação entre as cepas. A L variável e MBW foram altamente relacionada com H. argentinensis JIs. Três grupos foram separados por esta fase considerando PC1 e PC2: um formado por H. bacteriophora OLI, RIV e estirpes RN, (isolados de Córdoba e província de Rio Negro), um para a estirpe H. bacteriophora VELI (província de Buenos Aires) e um para H. argentinensis (província de Santa Fe). Heterorhabditis bacteriophora VELI e H. argentinensis isolado a partir de regiões com mais chuvas e umidade apresentaram maiores valores para as características morfométricas. A análise molecular mostrou as populações da Argentina (estirpe H. bacteriophora VELI e H. argentinensis), formando um mesmo subtipo, com seis outras populações H. bacteriophora (não da Argentina), com uma similaridade genética entre eles de 99%. Heterorhabditis argentinensis apresentado um único nucleótido que não estava presente em nenhum dos outros espécies do clado. Considerando os resultados deste estudo H. argentinensis seria conspecific a H. bacteriophora, constituindo uma estirpe com uma grande variação morfométrica onde o anfitrião e as condições climáticas podem ter influenciado nas medições.Fil: Achinelly, Maria Fernanda. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Estudios Parasitológicos y de Vectores. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Centro de Estudios Parasitológicos y de Vectores; ArgentinaFil: Eliceche, Daiana Pamela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Estudios Parasitológicos y de Vectores. Universidad Nacional de La Plata. Facultad de Ciencias Naturales y Museo. Centro de Estudios Parasitológicos y de Vectores; ArgentinaFil: Belaich, Mariano Nicolas. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular ; ArgentinaFil: Ghiringhelli, Pablo Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones en Ingeniería Genética y Biología Molecular ; Argentin

Virus como agentes de control biológico de poblaciones de mosquitos

Este capítulo trata sobre el viroma descripto para los mosquitos, con especial énfasis en el estudio de los patógenos que no afectan a los mamíferos. De este modo, el foco del capítulo es describir los virus de mosquitos que pueden ser utilizados como principios activos para la formulación de controladores biológicos. Se hace un relevamiento del estado del arte actual a nivel internacional sobre tales asuntos, y también se hace una revisión particular de la situación en Argentina.Fil: Belaich, Mariano Nicolas. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Ingeniería Genética y Biología Molecular y Celular. Área Virus de Insectos; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Claus, Juan Daniel. Universidad Nacional del Litoral. Facultad de Bioquímica y Ciencias Biológicas. Laboratorio de Virologia; Argentin

Effects of Fetal Bovine Serum deprivation in cell cultures on the production of Anticarsia gemmatalis Multinucleopolyhedrovirus

<p>Abstract</p> <p>Background</p> <p><it>Anticarsia gemmatalis </it>is a pest in South America's soybean crops, which could be controlled by the Multinucleopolyhedrovirus of <it>A. gemmatalis </it>(AgMNPV). Currently, its commercial production is based on infected larvae. However, the possibility of using modified baculoviruses in Integrated Pest Management programs has stimulated an interest to develop alternative multiplication processes. This study evaluated the AgMNPV production in UFL-Ag-286 cells previously deprived Fetal Bovine Serum.</p> <p>Results</p> <p>Culture media containing 1% FBS during the previous 48 hours achieved a synchronized condition where 90% of cells were found in G₀/G₁stage, showing the presence of non-filamentous actin. All characteristics were estimated from cellular viability tests, cell actin detection trials and flow cytometer cell cycle analysis. AgMNPV production was tested by transcript studies and budded viruses (BVs) and occlusion bodies (OBs) yield quantitation. Results showed that the productivity in FBS deprived cells was 9.8 times more in BVs and 3.8 times more in OBs with respect to non-treated cells.</p> <p>Conclusions</p> <p>UFL-Ag-286 cells previously deprived in FBS shown to be a better host for AgMNPV propagation, increasing the useful for both <it>in vitro </it>bioinsecticide production and applications such as recombinant protein expression or gene delivery.</p

Baculovirus: Molecular Insights on Their Diversity and Conservation

The Baculoviridae is a large group of insect viruses containing circular double-stranded DNA genomes of 80 to 180 kbp. In this study, genome sequences from 57 baculoviruses were analyzed to reevaluate the number and identity of core genes and to understand the distribution of the remaining coding sequences. Thirty one core genes with orthologs in all genomes were identified along with other 895 genes differing in their degrees of representation among reported genomes. Many of these latter genes are common to well-defined lineages, whereas others are unique to one or a few of the viruses. Phylogenetic analyses based on core gene sequences and the gene composition of the genomes supported the current division of the Baculoviridae into 4 genera: Alphabaculovirus, Betabaculovirus, Gammabaculovirus, and Deltabaculovirus

The ac53, ac78, ac101, and ac103 genes are newly discovered core genes in the family Baculoviridae

The family Baculoviridae is a large group of insect viruses containing circular double-stranded DNA genomes of 80 to 180 kbp, which have broad biotechnological applications. A key feature to understand and manipulate them is the recognition of orthology. However, the differences in gene contents and evolutionary distances among the known members of this family make it difficult to assign sequence orthology. In this study, the genome sequences of 58 baculoviruses were analyzed, with the aim to detect previously undescribed core genes because of their remote homology. A routine based on Multi PSI-Blast/tBlastN and Multi HaMStR allowed us to detect 31 of 33 accepted core genes and 4 orthologous sequences in the Baculoviridae which were not described previously. Our results show that the ac53, ac78, ac101 (p40), and ac103 (p48) genes have orthologs in all genomes and should be considered core genes. Accordingly, there are 37 orthologous genes in the family Baculoviridae.Fil: Garavaglia, Matías Javier. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Ingeniería Genética y Biología Molecular y Celular. Área Virus de Insectos; ArgentinaFil: Miele, Solange. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Ingeniería Genética y Biología Molecular y Celular. Área Virus de Insectos; ArgentinaFil: Iserte, Javier Alonso. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Ingeniería Genética y Biología Molecular y Celular. Área de Virosis Emergentes y Zoonótica; ArgentinaFil: Belaich, Mariano Nicolas. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Ingeniería Genética y Biología Molecular y Celular. Área Virus de Insectos; ArgentinaFil: Ghiringhelli, Pablo Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Ingeniería Genética y Biología Molecular y Celular. Área Virus de Insectos; Argentin

Isolation and Characterization of a Nucleopolyhedrovirus from Rachiplusia nu (Guenée) (Lepidoptera: Noctuidae)

Rachiplusia nu (Guenée) (Lepidoptera: Noctuidae) is an important pest of vegetable crops widely distributed in South America. At present, its control is based on the use of broad spectrum chemical insecticides. In order to develop a more selective alternative to be applied in integrated or organic pest management programs, we isolated and characterized a baculovirus that infects R. nu, denominated RanuMNPV (Rachiplusia nu multiple nucleopolyhedrovirus) in view of its morphology and pattern of infection. According to optical microscope images of infected in vitro cell cultures, data from bioassays carried out with various species of larvae, and DNA sequences from p74, polyhedrin, v-cathepsin, v-chitinase, lef8 and lef9 genes, this isolate could be considered as a variant of Autographa californica MNPV with a different host range.Fil: Rodriguez, Vanina Andrea. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Ingeniería Genética y Biología Molecular y Celular. Área Virus de Insectos; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Belaich, Mariano Nicolas. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Ingeniería Genética y Biología Molecular y Celular. Área Virus de Insectos; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Quintana, Graciela. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Microbiología y Zoología Agrícola; ArgentinaFil: Sciocco Cap, Alicia. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Microbiología y Zoología Agrícola; ArgentinaFil: Ghiringhelli, Pablo Daniel. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Ingeniería Genética y Biología Molecular y Celular. Área Virus de Insectos; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

Identification of nucleopolyhedrovirus that infect Nymphalid butterflies Agraulis vanillae and Dione juno

Dione juno and Agraulis vanillae are very common butterflies in natural gardens in South America, and also bred worldwide. In addition, larvae of these butterflies are considered as pests in crops of Passiflora spp.For these reasons, it is important to identify and describe pathogens of these species, both for preservation purposes and for use in pest control. Baculoviridae is a family of insect viruses that predominantly infect species of Lepidoptera and are used as bioinsecticides. Larvae of D. juno and A. vanillae exhibiting symptoms of baculovirus infection were examined for the presence of baculoviruses by PCR and transmission electron microscopy. Degenerate primers were designed and used to amplify partial sequences from the baculovirus p74, cathepsin, and chitinase genes, along with previously designed primers for amplification of lef-8, lef-9, and polh. Sequence data from these six loci, along with ultrastructural observations on occlusion bodies isolated from the larvae, confirmed that the larvae were infected with nucleopolyhedroviruses from genus Alphabaculovirus. The NPVs from the two different larval hosts appear to be variants of the same, previously undescribed baculovirus species. Phylogenetic analysis of the sequence data placed these NPVs in Alphabaculovirus group I/clade 1b.Fil: Rodriguez, Vanina Andrea. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Microbiología y Zoología Agrícola; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Ingeniería Genética y Biología Molecular y Celular. Área Virus de Insectos; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Belaich, Mariano Nicolas. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Microbiología y Zoología Agrícola; Argentina. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Ingeniería Genética y Biología Molecular y Celular. Área Virus de Insectos; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Mengual Gómez, Diego Luis. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Ingeniería Genética y Biología Molecular y Celular. Área Virus de Insectos; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Sciocco Cap, Alicia. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Microbiología y Zoología Agrícola; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Ghiringhelli, Pablo Daniel. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Ingeniería Genética y Biología Molecular y Celular. Área Virus de Insectos; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentin

RNA interference (RNAi): an effective tool in agrobiotechnology

El ARN de interferencia (ARNi) es un mecanismo evolutivamente conservado en la mayoría de las células eucariotas que permite silenciar genes mediante la degradación de ARN mensajero (ARNm) y la supresión de la síntesis de proteínas. En plantas, las moléculas de ARNi están involucradas en mecanismos de defensa contra patógenos y transposones, en la respuesta adaptativa al estrés, y en la expresión de genes relacionados con su crecimiento. El ARNi se considera una herramienta biotecnológica eficaz para silenciar la expresión de genes de microorganismos fitopatógenos, esto permite el diseño de bioplaguicidas ambientalmente seguros con una afinidad y selectividad, en muchos casos superior a la de los plaguicidas químicos. En esta revisión se señalan los últimos avances en la aplicación del ARNi en el contexto agrícola y su efectividad en el control biológico de fitopatógenos e insectos plaga. Asimismo, se presentan diversos ensayos experimentales cuyos resultados pueden ser la base para futuros bioproductos, además de algunos ejemplos disponibles en el mercado. Por último, se abordan aspectos de bioseguridad y consideraciones regulatorias necesarias para la aceptación y uso de esta tecnología a nivel global.RNA interference (RNAi) is an evolutionarily conserved mechanism in most eukaryotic cells that allows genes to be silenced by degradation of messenger RNA (mRNA) and suppression of protein synthesis. In plants, RNAi molecules are involved in defense mechanisms against pathogens and transposons, in the adaptive response to stress, and in the expression of genes related to their growth. RNAi is an effective biotechnological tool to silence the expression of specific genes which are essential for the survival of phytopathogenic microorganisms, thus allowing the design of environmentally safe biopesticides with affinity and selectivity, in many cases greater than chemical pesticides. This review describes the latest advances in the application of RNAi in the agricultural context and its effectiveness in the biological control of phytopathogens and pest insects. Likewise, various experimental trials are presented, the results of which may be the basis for future bioproducts, as well as some examples available on the market. Finally, biosafety aspects and regulatory considerations necessary for the acceptance and use of this technology at a global level are presented.Fil: Gamero, Maira. Corporación Colombiana de Investigación Agropecuaria; ColombiaFil: Tolosa Moreno, Deisy. Corporación Colombiana de Investigación Agropecuaria; ColombiaFil: Belaich, Mariano Nicolas. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología. Laboratorio de Ingeniería Genética y Biología Molecular y Celular. Área Virus de Insectos; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Barrera, Gloria. Corporación Colombiana de Investigación Agropecuaria; Colombi