134 results on '"Sciocco-Cap, A."'
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2. Genomic analysis of an Argentinean isolate of Spodoptera frugiperda granulovirus reveals that various baculoviruses code for Lef-7 proteins with three F-box domains.
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María Leticia Ferrelli, Matías Luis Pidre, Pablo Daniel Ghiringhelli, Sofía Torres, María Laura Fabre, Tomás Masson, Maia Tatiana Cédola, Alicia Sciocco-Cap, and Víctor Romanowski
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Medicine ,Science - Abstract
A new isolate of the Spodoptera frugiperda granulovirus, SfGV ARG, was completely sequenced and analyzed. The SfGV ARG genome is 139,812 bp long and encodes 151 putative open reading frames. Of these ORFs, 56 were found in betabaculoviruses, 19 of which are present only in GVs closely related to SfGV. Seven ORFs found homologs in this small GV group and also in noctuid NPVs. ORF066 codes a 74 amino acid protein, overlapped with nudix gene, with several homologs in baculovirus, found by tblastn search. Comparison with the genome of the Colombian isolate SfGV VG008 resulted in SfGV being 1101 bp smaller and lacking a homologue of VG008 ORF084, which codes for Lef-7. However, we found that ORF051 shows remote homology to Lef-7 proteins. Moreover, analysis of ORF051 along with Lef-7 proteins coded by a group of noctuid specific GVs and NPVs indicated that Lef-7 proteins coded by these viruses include three F-box domains in contrast to the single one reported for AcMNPV Lef-7. SfGV ARG genome also contains a split photolyase as a distinct feature not found in VG008. BlastX analysis revealed that a complete photolyase is coded considering a putative frameshift in a poly-A tract, which resembles known slippery sequences involved in programmed ribosome frameshifting.
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- 2018
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3. Baculovirus Insecticides in Latin America: Historical Overview, Current Status and Future Perspectives
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Santiago Haase, Alicia Sciocco-Cap, and Víctor Romanowski
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viral biopesticides ,baculovirus ,Latin America ,crop protection ,Microbiology ,QR1-502 - Abstract
Baculoviruses are known to regulate many insect populations in nature. Their host-specificity is very high, usually restricted to a single or a few closely related insect species. They are amongst the safest pesticides, with no or negligible effects on non-target organisms, including beneficial insects, vertebrates and plants. Baculovirus-based pesticides are compatible with integrated pest management strategies and the expansion of their application will significantly reduce the risks associated with the use of synthetic chemical insecticides. Several successful baculovirus-based pest control programs have taken place in Latin American countries. Sustainable agriculture (a trend promoted by state authorities in most Latin American countries) will benefit from the wider use of registered viral pesticides and new viral products that are in the process of registration and others in the applied research pipeline. The success of baculovirus-based control programs depends upon collaborative efforts among government and research institutions, growers associations, and private companies, which realize the importance of using strategies that protect human health and the environment at large. Initiatives to develop new regulations that promote the use of this type of ecological alternatives tailored to different local conditions and farming systems are underway.
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- 2015
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4. Development of a Recombination System for the Generation of Occlusion Positive Genetically Modified Anticarsia Gemmatalis Multiple Nucleopolyhedrovirus
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Santiago Haase, Christina B. McCarthy, M. Leticia Ferrelli, Matias L. Pidre, Alicia Sciocco-Cap, and Victor Romanowski
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AgMNPV ,recombinant baculovirus ,bioinsecticide ,Anticarsia gemmatalis ,velvetbean caterpillar ,Microbiology ,QR1-502 - Abstract
Anticarsia gemmatalis is an important pest in legume crops in South America and it has been successfully controlled using Anticarsia gemmatalis Multiple Nucleopolyhedrovirus (AgMNPV) in subtropical climate zones. Nevertheless, in temperate climates its speed of kill is too slow. Taking this into account, genetic modification of AgMNPV could lead to improvements of its biopesticidal properties. Here we report the generation of a two-component system that allows the production of recombinant AgMNPV. This system is based on a parental AgMNPV in which the polyhedrin gene (polh) was replaced by a bacterial β-galactosidase (lacZ) gene flanked by two target sites for the homing endonuclease I-PpoI. Co-transfection of insect cells with linearized (I-PpoI-digested) parental genome and a transfer vector allowed the restitution of polh and the expression of a heterologous gene upon homologous recombination, with a low background of non-recombinant AgMNPV. The system was validated by constructing a recombinant occlusion-positive (polh+) AgMNPV expressing the green fluorescent protein gene (gfp). This recombinant virus infected larvae normally per os and led to the expression of GFP in cell culture as well as in A. gemmatalis larvae. These results demonstrate that the system is an efficient method for the generation of recombinant AgMNPV expressing heterologous genes, which can be used for manifold purposes, including biotechnological and pharmaceutical applications and the production of orally infectious recombinants with improved biopesticidal properties.
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- 2015
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5. Analysis of a chitinase from EpapGV, a fast killing betabaculovirus
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Salvador, Ricardo, Ferrelli, M. Leticia, Sciocco-Cap, Alicia, and Romanowski, Víctor
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- 2014
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6. Prospection and molecular analysis of CpGV isolates infecting Cydia pomonella at different geographical locations in Argentina
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Arneodo, J. D., De Anna, J., Salvador, R., Farinon, M., Quintana, G., and Sciocco-Cap, A.
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- 2015
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7. Functional analysis of Spodoptera frugiperda nucleopolyhedrovirus late expression factors in Sf9 cells
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Berretta, Marcelo F., López, M. Gabriela, Taboga, Oscar, Sciocco-Cap, Alicia, and Romanowski, Víctor
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- 2013
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8. Analysis of EpapGV gp37 gene reveals a close relationship between granulovirus and entomopoxvirus
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Salvador, Ricardo, Ferrelli, M. Leticia, Berretta, Marcelo F., Mitsuhashi, Wataru, Biedma, Marina E., Romanowski, Víctor, and Sciocco-Cap, Alicia
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- 2012
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9. Detection and kinetic analysis of Epinotia aporema granulovirus in its lepidopteran host by real-time PCR
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Arneodo, J. D., König, G. A., Berretta, M. F., Di Rienzo, J. A., Taboga, O., and Sciocco-Cap, A.
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- 2012
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10. Multiplex PCR and quality control of Epinotia aporema granulovirus production
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Manzán, Maria Alejandra, Aljinovic, Ernesto Mario, Biedma, Marina Elizabeth, Sciocco-Cap, Alicia, Ghiringhelli, Pablo Daniel, and Romanowski, Víctor
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- 2008
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11. Sequencing and Characterisation of p74 Gene in Two Isolates of Anticarsia Gemmatalis MNPV
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Belaich, Mariano Nicolás, Rodríguez, Vanina Andrea, Bilen, Marcos Fabián, Pilloff, Marcela Gabriela, Romanowski, Victor, Sciocco-Cap, Alicia, and Ghiringhelli, Pablo Daniel
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- 2006
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12. The Baculoviral Genome
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Leticia, M., primary, F., Marcelo, additional, N., Mariano, additional, Daniel, P., additional, Sciocco-Cap, Alicia, additional, and Romanowski, Vctor, additional
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- 2012
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13. Physical and Genetic Map of Epinotia aporema Granulovirus Genome
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Parola, Alejandro D., Alejandra ManzÁn, M., Lozano, Mario E., Ghiringhelli, P. Daniel, Sciocco-Cap, Alicia, and Romanowski, VÍctor
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- 2002
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14. Identification and Characterization of the Ecdysteroid UDP-Glycosyltransferase Gene of Epinotia aporema Granulovirus
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Alejandra Manzán, María, Lozano, Mario E., Sciocco-Cap, Alicia, Ghiringhelli, Pablo D., and Romanowski, Víctor
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- 2002
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15. Characterization of a granulovirus isolated from Epinotia aporema Wals. (Lepidoptera: Tortricidae) larvae
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Sciocco-Cap, Alicia, Parola, Alejandro D., Goldberg, Alina V., Ghiringhelli, Pablo D., and Romanowski, Victor
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Insect pests -- Biological control ,Insect viruses -- Research ,Biological sciences - Abstract
The properties of a granulovirus that could be used in the biological control of the soybean shoot borer Epinotia aporema are described. The virus produces a protein that is homologous to other viral granulins.
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- 2001
16. Identification and Characterization of the Ecdysteroid UDP-Glycosyltransferase Gene of Epinotia aporema Granulovirus*
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Manzán, María Alejandra, Lozano, Mario E., Sciocco-cap, Alicia, Ghiringhelli, Pablo D., and Romanowski, Víctor
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- 2002
17. Genome of Epinotia aporema granulovirus (EpapGV), a polyorganotropic fast killing betabaculovirus with a novel thymidylate kinase gene
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Ferrelli María, Salvador Ricardo, Biedma Marina, Berretta Marcelo, Haase Santiago, Sciocco-Cap Alicia, Ghiringhelli Pablo, and Romanowski Víctor
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Biotechnology ,TP248.13-248.65 ,Genetics ,QH426-470 - Abstract
Abstract Background Epinotia aporema (Lepidoptera: Tortricidae) is an important pest of legume crops in South America. Epinotia aporema granulovirus (EpapGV) is a baculovirus that causes a polyorganotropic infection in the host larva. Its high pathogenicity and host specificity make EpapGV an excellent candidate to be used as a biological control agent. Results The genome of Epinotia aporema granulovirus (EpapGV) was sequenced and analyzed. Its circular double-stranded DNA genome is 119,082 bp in length and codes for 133 putative genes. It contains the 31 baculovirus core genes and a set of 19 genes that are GV exclusive. Seventeen ORFs were unique to EpapGV in comparison with other baculoviruses. Of these, 16 found no homologues in GenBank, and one encoded a thymidylate kinase. Analysis of nucleotide sequence repeats revealed the presence of 16 homologous regions (hrs) interspersed throughout the genome. Each hr was characterized by the presence of 1 to 3 clustered imperfect palindromes which are similar to previously described palindromes of tortricid-specific GVs. Also, one of the hrs (hr4) has flanking sequences suggestive of a putative non-hr ori. Interestingly, two more complex hrs were found in opposite loci, dividing the circular dsDNA genome in two halves. Gene synteny maps showed the great colinearity of sequenced GVs, being EpapGV the most dissimilar as it has a 20 kb-long gene block inversion. Phylogenetic study performed with 31 core genes of 58 baculoviral genomes suggests that EpapGV is the baculovirus isolate closest to the putative common ancestor of tortricid specific betabaculoviruses. Conclusions This study, along with previous characterization of EpapGV infection, is useful for the better understanding of the pathology caused by this virus and its potential utilization as a bioinsecticide.
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- 2012
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18. Biological and molecular features of Nosema rachiplusiae sp. n., a microsporidium isolated from the neotropical moth Rachiplusia nu (Guenée) (Lepidoptera: Noctuidae)
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Alicia Sciocco-Cap and J. D. Arneodo
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0106 biological sciences ,0301 basic medicine ,Argentina ,Zoology ,Moths ,01 natural sciences ,Lepidoptera genitalia ,03 medical and health sciences ,Rachiplusia nu ,Nosema ,Agronomía, reproducción y protección de plantas ,parasitic diseases ,Animals ,DNA, Fungal ,Phylogeny ,Microscopy ,General Veterinary ,biology ,fungi ,General Medicine ,Spores, Fungal ,biology.organism_classification ,Microsporidium ,010602 entomology ,030104 developmental biology ,Infectious Diseases ,RNA, Ribosomal ,CIENCIAS AGRÍCOLAS ,Insect Science ,Larva ,Microsporidia ,Ultrastructure ,Noctuidae ,Parasitology ,Polar filament ,Agricultura, Silvicultura y Pesca ,Ribosomes - Abstract
Light, electron microscopy and DNA analyses were performed to characterize a microsporidium infecting Rachiplusia nu larvae from a laboratory rearing in Argentina. Diplokaryotic spores were oval and measured 3.61 ± 0.29 × 1.61 ± 0.14 μM (fresh). The spore wall was composed of an electron-dense exospore and an electron-lucent endospore, ca. 30 nm and 100–120 nm thick, respectively. The polar filament was arranged in a single rank of 10–12 coils (typically 11). Microsporidian cells were found in the cytoplasm, next to the endoplasmic reticulum (especially the prespore stages) and generally surrounded by electron-lucent spaces. The infection was polyorganotropic; the fat body appeared as the most heavily invaded tissue, followed by tracheal matrix and epidermis. A molecular phylogeny based on the small (SSU) and large subunit (LSU) ribosomal RNA genes clearly placed the new isolate within the “Nosema bombycis clade”. Considering both SSU and LSU concatenated partial sequences, the microsporidium from R. nu showed 99.5% nucleotide similarity with N. bombycis and 99.8% with its closest relative, a microsporidium isolated from Philosamia cynthia. According to its genetic and biological features, the R. nu isolate is proposed as the new species Nosema rachiplusiae sp. n., expanding the limited knowledge on microsporidia associated to endemic South-American moths. Fil: Arneodo Larochette, Joel Demián. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Microbiología y Zoología Agrícola; Argentina Fil: Sciocco Cap, Alicia. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Microbiología y Zoología Agrícola; Argentina
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- 2018
19. Biological and molecular characterization of Rachiplusia nu single nucleopolyhedrovirus, a promising biocontrol agent against the South American soybean pest Rachiplusia nu
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Jakubowicz, Violeta, primary, Taibo, Catalina B., additional, Sciocco-Cap, Alicia, additional, and Arneodo, Joel D., additional
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- 2019
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20. Biology and morphometrics of the immature stages of Epinotia aporema on artificial diet Biologia e morfometria dos estágios imaturos de Epinotia aporema em dieta artificial
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Joel Arneodo, Graciela Quintana, and Alicia Sciocco-Cap
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Glycine max ,broca-das-axilas ,cápsula cefálica ,cria em laboratório ,larvas ,bean shoot borer ,cephalic capsule ,laboratory rearing ,larvae ,Agriculture (General) ,S1-972 - Abstract
The objective of this work was to establish a life table for the immature stages of Epinotia aporema, as part of a wider investigation on its biological control. Insects were reared on an artificial diet at 25±1ºC and a 16:8 (light:dark) hour photoperiod. For the identification of larval instars for the study of pathogen-insect interactions under laboratory conditions, head capsule widths (HCWs) were also determined. The egg incubation period was 4.13±0.30 days, larval stage took 11.64±0.49 days, and the development time of the pupal phase was sex-dependent with 8.51±0.69 days for the females and 9.41±0.65 days for the males. Five larval instars were identified.O objetivo deste trabalho foi estabelecer uma tabela de vida para os estágios imaturos de Epinotia aporema, como parte de um estudo mais amplo para seu controle biológico. Os insetos foram criados em dieta artificial a 25±1ºC e 16:8 (luz:escuridão) horas de fotoperíodo. Para a identificação dos estágios larvais para estudos de interação inseto-patógeno em condições de laboratório, as larguras de cápsula cefálica também foram determinadas. O período de incubação dos ovos foi de 4,13±0,30 dias, o estágio larval foi de 11,64±0,49 dias, e o tempo de desenvolvimento das pupas dependeu do sexo, com 8,51±0,69 dias para as fêmeas e 9,41±0,65 dias para os machos. Foram identificados cinco estágios larvais.
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- 2010
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21. Prospection and molecular analysis of CpGV isolates infectingCydia pomonellaat different geographical locations in Argentina
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Alicia Sciocco-Cap, G. Quintana, J. D. Arneodo, J. De Anna, R. Salvador, and M. Farinon
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Lepidoptera genitalia ,Veterinary medicine ,Genetic diversity ,Biopesticide ,Larva ,Phylogenetic tree ,Botany ,Cydia pomonella ,Biology ,Agronomy and Crop Science ,Cydia pomonella granulovirus ,Molecular analysis - Abstract
The occurrence and genetic diversity of Cydia pomonella granulovirus infecting C. pomonella across Argentina were investigated. Larvae were collected from fruit orchards in regions where CpGV-based biopesticides have been extensively used and then discontinued (Catamarca) or are still being used (Alto Valle). The survey was also extended to regions where few occasional (i.e. Mendoza) or no virus treatments (various locations) have been made. PCR analysis showed an unexpectedly high proportion of infected larvae (31.3–50%) at most sampling sites in Catamarca. Phylogenetic analysis of these samples, based on gran and lef-8 genes partial sequences, revealed a previously unreported CpGV variant which could not be assigned to any of the known CpGV genome types and formed a well-defined new cluster. Instead, samples from Alto Valle were genetically similar to isolate CpGV-M (the active ingredient in commercial formulations). There, the percentage of infected individuals was considerably lower (1.8–21.9%). The virus was not detected in Mendoza or the other sampling regions. Implications of these results on the epidemiology and current classification of CpGV isolates are discussed.
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- 2014
22. Biological and molecular characterization of Rachiplusia nu single nucleopolyhedrovirus, a promising biocontrol agent against the South American soybean pest Rachiplusia nu
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Alicia Sciocco-Cap, Catalina Taibo, Joel D. Arneodo, and Violeta Jakubowicz
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0106 biological sciences ,0301 basic medicine ,animal structures ,biology ,fungi ,Biological pest control ,Plusiinae ,Moths ,Spodoptera ,biology.organism_classification ,Diatraea saccharalis ,01 natural sciences ,Nucleopolyhedroviruses ,Microbiology ,Lepidoptera genitalia ,010602 entomology ,03 medical and health sciences ,030104 developmental biology ,Rachiplusia nu ,Animals ,Noctuidae ,PEST analysis ,Pest Control, Biological ,Ecology, Evolution, Behavior and Systematics - Abstract
Rachiplusia nu (Lepidoptera: Noctuidae) is a key soybean pest in Argentina. Current management of this moth relies mainly on the use of synthetic insecticides and transgenic plants. In search of biological control-based alternatives, a baculovirus from R. nu (hereafter RanuNPV) was characterized and its insecticidal properties tested under laboratory conditions. RanuNPV occlusion bodies (OBs) were nearly tetrahedral, averaging 1.0 ± 0.2 µm in their longest edge and containing singly enveloped nucleocapsids. Histopathology of infected late-instar larvae revealed broad tissue tropism, where fat bodies and epidermis were the most affected organs. Phylogenetic analysis of concatenated polh, lef-8 and lef-9 partial sequences classified RanuNPV as a new species that clusters with other group II alphabaculoviruses infecting larvae of Plusiinae. Bioassays performed with R. nu neonates determined the median lethal dosage to be approximately 2.5 OBs/larva; most insects died within 4–5 days post inoculation showing typical baculovirus-induced liquefaction. No effects were observed in other lepidopteran species assayed, including Spodoptera frugiperda, Cydia pomonella and Diatraea saccharalis. High pathogenicity and host specificity make RanuNPV a good candidate for controlling R. nu.
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- 2019
23. Genomic analysis of an Argentinean isolate of Spodoptera frugiperda granulovirus reveals that various baculoviruses code for Lef-7 proteins with three F-box domains
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Ferrelli, María Leticia, primary, Pidre, Matías Luis, additional, Ghiringhelli, Pablo Daniel, additional, Torres, Sofía, additional, Fabre, María Laura, additional, Masson, Tomás, additional, Cédola, Maia Tatiana, additional, Sciocco-Cap, Alicia, additional, and Romanowski, Víctor, additional
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- 2018
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24. Characterization of a new Helicoverpa armigera nucleopolyhedrovirus variant causing epizootic on a previously unreported host, Helicoverpa gelotopoeon (Lepidoptera: Noctuidae)
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M.L. Ferrelli, Catalina Taibo, J.D. Arneodo, P. Fichetti, and Alicia Sciocco-Cap
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0106 biological sciences ,0301 basic medicine ,HELICOVERPA GELOTOPOEON ,Genes, Viral ,Biología ,viruses ,Otras Ciencias Biológicas ,Biology ,Moths ,01 natural sciences ,Polymerase Chain Reaction ,Virus ,Lepidoptera genitalia ,purl.org/becyt/ford/1 [https] ,Ciencias Biológicas ,03 medical and health sciences ,Nucleopolyhedrovirus ,Electron microscopy ,medicine ,Animals ,PHYLOGENETIC ANALYSIS ,NUCLEOPOLYHEDROVIRUS ,purl.org/becyt/ford/1.6 [https] ,Ecology, Evolution, Behavior and Systematics ,Epizootic ,Phylogeny ,Phylogenetic analysis ,Phylogenetic tree ,Base Sequence ,Host (biology) ,fungi ,biology.organism_classification ,medicine.disease ,Virology ,Nucleopolyhedroviruses ,010602 entomology ,ELECTRON MICROSCOPY ,030104 developmental biology ,Virus Diseases ,Noctuidae ,Helicoverpa gelotopoeon ,PEST analysis ,CIENCIAS NATURALES Y EXACTAS - Abstract
This paper reports the first biological and molecular characterization of a nucleopolyhedrovirus isolated from the soybean and cotton pest Helicoverpa gelotopoeon. Studies were performed following a virus outbreak in a rearing facility and in wild H. gelotopoeon populations in Córdoba, Argentina. Host identity was corroborated by partial sequencing of the COI gene. Scanning electron microscope observations of purified OBs revealed their polyhedral morphology and an average diameter of 0.89 ± 0.14 μm. Ultrathin sections of infected larvae examined by transmission electron microscopy showed the intranuclear occurrence of polyhedra and virus particles in fat body cells. Nucleocapsids were singly enveloped. Phylogenetic analysis of lef-8, lef-9, polh, orf5/5b and hr3-orf62 viral sequences identified this new NPV isolate (hereafter HegeSNPV) as a variant of Helicoverpa armigera nucleopolyhedrovirus (HearNPV). Furthermore, HegeSNPV was closely related to the so-called “HzSNPV Group” within HearNPV, although having particular characteristics., Instituto de Biotecnologia y Biologia Molecular
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- 2016
25. Light microscopy of microsporidian spores and baculovirus occlusion bodies in Epinotia aporema and Helicoverpa gelotopoeon larval tissues, respectively
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Catalina Taibo, Alicia Sciocco-Cap, and Joel D. Arneodo
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0301 basic medicine ,Microbiology (medical) ,Pathology ,medicine.medical_specialty ,Moths ,Epinotia aporema ,Microbiology ,Inclusion bodies ,Inclusion Bodies, Viral ,Microscopia ,03 medical and health sciences ,Nosema ,Microscopy ,medicine ,Animals ,Plagas de plantas ,Larva ,biology ,Staining and Labeling ,Epinotia ,General Medicine ,Spores, Fungal ,biology.organism_classification ,Pest of Plants ,Entomopatógenos ,Nucleopolyhedroviruses ,Spore ,030104 developmental biology ,Organ Specificity ,Helicoverpa ,Helicoverpa gelotopoeon - Abstract
Entomopathogens help regulate natural insect populations, and their use in pest management is the subject of active research. Classical and molecular techniques are currently available to elucidate disease etiology and pathogenesis. Here, we performed an Azan trichrome stain for light microscopic visualization of pathogens in the larval tissues of two Neotropical lepidopterans. Epinotia aporema larvae exhibiting symptoms of microsporidiosis were fortuitously detected in a rearing facility (IMyZA-INTA, Argentina), while Helicoverpa gelotopoeon specimens were experimentally infected with an Argentinean isolate of Helicoverpa armigera nucleopolyhedrovirus (HearNPV). Caterpillars were fixed in Duboscq-Brasil and paraffin embedded. Microtome sections (5 μm-thick) were deparaffinized and stained2 for observation under a Nikon Eclipse 80i microscope equipped with a Nikon DS-U3 digital camera. Inst. de Microbiología y Zoología Agrícola IMyZA Fil: Arneodo Larochette, Joel Demian. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Microbiología y Zoología Agrícola; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Taibo, Catalina Beatriz. Instituto Nacional de Tecnología Agropecuaria (INTA). Centro de Investigación en Ciencias Veterinarias y Agronómicas. Servicio de Microscopía; Argentina Fil: Sciocco, Alicia Ines. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Microbiología y Zoología Agrícola; Argentina
- Published
- 2016
26. Detection and kinetic analysis of Epinotia aporema granulovirus in its lepidopteran host by real-time PCR
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J. D. Arneodo, Alicia Sciocco-Cap, Oscar Taboga, Marcelo Facundo Berretta, J. A. Di Rienzo, and Guido König
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Baculoviridae ,biology ,Epinotia ,Host (biology) ,Biological pest control ,General Medicine ,Real-Time Polymerase Chain Reaction ,biology.organism_classification ,Virology ,Virus ,Lepidoptera ,Lepidoptera genitalia ,Kinetics ,Viral Proteins ,Titer ,Animals ,PEST analysis - Abstract
Epinotia aporema granulovirus (EpapGV) has attracted interest as a potential biocontrol agent of the soybean pest Epinotia aporema in Argentina. Studies on virus/host interactions conducted so far have lacked an accurate method to assess the progress of virus load during the infection process. The present paper reports the development of a real-time PCR for EpapGV and its application to describe viral kinetics following ingestion of two different virus doses by last-instar E. aporema larvae. Real-time PCR was shown to be a reliable method to detect and quantify the presence of EpapGV in the analyzed samples. The increase in virus titer (log) exhibited a sigmoidal pattern, with an exponential growth phase between 24 and 48 h postinfection for both initial doses tested.
- Published
- 2012
27. Molecular Identification of Helicoverpa armigera (Lepidoptera: Noctuidae: Heliothinae) in Argentina and Development of a Novel PCR-RFLP Method for its Rapid Differentiation From H. zea and H. gelotopoeon
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Fernando Miguel Flores, Alicia Sciocco-Cap, Emilia Ines Balbi, and Joel D. Arneodo
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Male ,Otras Ciencias Biológicas ,Argentina ,Zoology ,Heliothinae ,Helicoverpa armigera ,Moths ,Polymerase Chain Reaction ,Lepidoptera genitalia ,Electron Transport Complex IV ,Ciencias Biológicas ,Botany ,Animals ,Helicoverpa ,Barcode ,Ecology ,biology ,fungi ,food and beverages ,General Medicine ,South America ,biology.organism_classification ,Phylogeography ,Insect Science ,Noctuidae ,Helicoverpa gelotopoeon ,Helicoverpa zea ,Old world bollworm ,Restriction fragment length polymorphism ,Biological invasion ,Polymorphism, Restriction Fragment Length ,CIENCIAS NATURALES Y EXACTAS - Abstract
Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae: Heliothinae) is among the most voracious global pests of agriculture. Adults of this species were identified recently in northern Argentina by dissection of male genitalia. In this work, a rapid and simple molecular tool was designed to distinguish H. armigera from the morphologically similar indigenous bollworms Helicoverpa zea (Boddie) and Helicoverpa gelotopoeon (Dyar), regardless of the life stage. Amplification of partial COI gene with a new primer pair, and subsequent digestion with endonuclease HinfI, yielded different RFLP profiles for the three main Helicoverpa pests currently present in South America. The method was validated in Helicoverpa specimens collected across Argentina, whose identity was further corroborated by COI sequencing and phylogenetic analysis. The data reported here constitute the first molecular confirmation of thispest in the country. The survey revealed the occurrence of H. armigera in northern and central Argentina, including the main soybean- and maize-producing area. Fil: Arneodo Larochette, Joel Demián. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Microbiología y Zoología Agrícola; Argentina Fil: Balbi, Emilia I.. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Córdoba. Estación Experimental Agropecuaria Marcos Juárez; Argentina Fil: Flores, Fernando M.. Instituto Nacional de Tecnología Agropecuaria. Centro Regional Córdoba. Estación Experimental Agropecuaria Marcos Juárez; Argentina Fil: Sciocco Cap, Alicia. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Microbiología y Zoología Agrícola; Argentina
- Published
- 2015
28. Effect of the interaction between Anticarsia gemmatalis multiple nucleopolyhedrovirus and Epinotia aporema granulovirus, on A. gemmatalis (Lepidoptera: Noctuidae) larvae
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Ricardo Salvador, Alicia Sciocco-Cap, Víctor Romanowski, Marina Elizabeth Biedma, and M. Leticia Ferrelli
- Subjects
Tortricidae ,Veterinary medicine ,Epinotia aporema granulovirus ,animal structures ,Otras Ciencias Biológicas ,Granulovirus ,Soybean pest ,Lepidoptera genitalia ,Ciencias Biológicas ,Control Biológico ,Botany ,soja ,Baculovirus ,Caterpillar ,Larva ,Glycine Max ,biology ,Anticarsia gemmatalis nucleopolyhedrovirus ,fungi ,food and beverages ,biology.organism_classification ,Anticarsia gemmatalis ,Membrana Peritrófica ,Insect Science ,Biological control ,Epinotia Aporema ,Peritrophic membrane ,Noctuidae ,Instar ,Soybeans ,Agronomy and Crop Science ,CIENCIAS NATURALES Y EXACTAS - Abstract
The bean shoot borer Epinotia aporema Wals. (Lepidoptera: Tortricidae) and the velvet bean caterpillar Anticarsia gemmatalis Hübner (Lepidoptera: Noctuidae) are key pests of soybean and other legume crops in South America. They are often found simultaneously in certain regions. A. gemmatalis nucleopolyhedrovirus (AgMNPV) is widely used to control A. gemmatalis. More recently, E. aporema granulovirus (EpapGV) has been characterized and evaluated as a bioinsecticide for E. aporema. In order to increase its potential use and to design optimized strategies for the management of lepidopteran pests, we evaluated the interaction between EpapGV and AgMNPV on third instar A. gemmatalis larvae. Larvae fed with 50 AgMNPV OBs/larva showed an increase in the mortality rates (from 42% to 81%) and a decrease in the median survival time (from 7.7 days to 5.7 days) when these OBs were mixed with 6000 EpapGV OBs/larva. When 300 AgMNPV OBs/larva were used alone or in combination with EpapGV OBs no changes in biological parameters were observed. No mortality was detected in A. gemmatalis larvae treated with EpapGV alone. In larvae fed with the viral mixtures, only AgMNPV DNA was detected by PCR. A. gemmatalis peritrophic membranes (PMs) examined by SDS–PAGE and scanning electron microscopy showed signs of damage. Notably, we found the presence of spheroidal bodies associated with damaged areas in the PMs of larvae fed with EpapGV but not in those that were given AgMNPV alone. These results show that EpapGV increases the viral potency of AgMNPV, and thus the insecticidal efficiency, suggesting that the use of formulations including both viruses might be a valuable tool for pest management. Fil: Biedma, Marina Elizabeth. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Biotecnología y Biología Molecular. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Instituto de Biotecnología y Biología Molecular; Argentina Fil: Salvador, Ricardo. 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. Centro Científico Tecnológico Conicet - La Plata. Instituto de Biotecnología y Biología Molecular. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Instituto de Biotecnología y Biología Molecular; Argentina Fil: Ferrelli, Maria Leticia. 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. Centro Científico Tecnológico Conicet - La Plata. Instituto de Biotecnología y Biología Molecular. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Instituto de Biotecnología y Biología Molecular; Argentina Fil: Sciocco Cap, Alicia. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Microbiología y Zoología Agrícola; Argentina Fil: Romanowski, Victor. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Biotecnología y Biología Molecular. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Instituto de Biotecnología y Biología Molecular; Argentina
- Published
- 2015
29. Identification of nucleopolyhedrovirus that infect Nymphalid butterflies Agraulis vanillae and Dione juno
- Author
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Pablo Daniel Ghiringhelli, Mariano Nicolás Belaich, Vanina Andrea Rodríguez, Diego Luis Mengual Gómez, and Alicia Sciocco-Cap
- Subjects
Baculoviridae ,animal structures ,food.ingredient ,media_common.quotation_subject ,Zoology ,Insect ,Lepidoptera genitalia ,food ,Phylogenetics ,Botany ,Polyhedrin ,Animals ,Pest Control, Biological ,Ecology, Evolution, Behavior and Systematics ,media_common ,Inclusion Bodies ,biology ,Phylogenetic tree ,fungi ,South America ,biology.organism_classification ,Nucleopolyhedroviruses ,Alphabaculovirus ,Larva ,DNA, Viral ,Butterflies ,Dione juno - Abstract
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.
- Published
- 2011
30. Multiplex PCR and quality control of Epinotia aporema granulovirus production
- Author
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Alicia Sciocco-Cap, Marina Elizabeth Biedma, Maria Alejandra Manzan, Ernesto M. Aljinovic, Võ ´ ctor Romanowski, and Pablo Daniel Ghiringhelli
- Subjects
Genetics ,Virus Cultivation ,Epinotia aporema granulovirus ,Granulovirus ,General Medicine ,Moths ,Biology ,Virus Replication ,Polymerase Chain Reaction ,Sensitivity and Specificity ,Genome ,DNA sequencing ,chemistry.chemical_compound ,chemistry ,Virology ,DNA, Viral ,Multiplex polymerase chain reaction ,Animals ,Genomic library ,Primer (molecular biology) ,Molecular Biology ,DNA ,DNA Primers - Abstract
A specific multiplex PCR was developed for the rapid and highly sensitive quality control of the viral DNA during Epinotia aporema granulovirus (EpapGV) production. At the beginning of this work only 2.3% of the EpapGV genomic sequence was known. In order to increase the availability of specific information, the ter- minal sequences of the inserts of several selected clones of EpapGV genomic libraries were determined. These data comprised 8.4% of the total DNA sequence and corre- sponded to regions distributed throughout the genome. Based on the small fraction of known sequence available a set of 32 primers was designed, using information theory to set the basis for this study. Each pair of designed primers was initially tested in individual PCRs to assess the correct size of the expected product and the sensitivity of the amplification. The specificity was verified in multiplex PCRs, using alternatively 1-3 sets of selected 5-6 primer pairs and EpapGV DNA preparations from different sour- ces and degrees of purity. The results indicate that the multiplex PCR could be used for quality control in the bioinsecticide production, as well as in other applications such as the detection of latent infections in E. aporema colonies, and studies related to virus distribution, vertical transmission, host range, or persistence in the field.
- Published
- 2008
31. Physical and Genetic Map of Epinotia aporema Granulovirus Genome
- Author
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Parola, Alejandro D., Alejandra ManzÁn, M., Lozano, Mario E., Ghiringhelli, P. Daniel, Sciocco-Cap, Alicia, and Romanowski, VÍctor
- Published
- 2003
- Full Text
- View/download PDF
32. Light microscopy of microsporidian spores and baculovirus occlusion bodies in Epinotia aporema and Helicoverpa gelotopoeon larval tissues, respectively
- Author
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Arneodo, Joel D., primary, Taibo, Catalina, additional, and Sciocco-Cap, Alicia, additional
- Published
- 2016
- Full Text
- View/download PDF
33. Prospection and molecular analysis of CpGV isolates infecting Cydia pomonella at different geographical locations in Argentina
- Author
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Arneodo Larochette, Joel Demián, De Anna, J., Salvador, R., Farinon, M., Quintana, G., and Sciocco Cap, A.
- Subjects
Ciencias Biológicas ,CYDIA POMONELLA ,CPGV ,Otras Ciencias Biológicas ,EPIDEMIOLOGY ,GENETIC DIVERSITY ,BACULOVIRIDAE ,LEPIDOPTERA ,CIENCIAS NATURALES Y EXACTAS - Abstract
The occurrence and genetic diversity of Cydia pomonella granulovirus infecting C. pomonella across Argentina were investigated. Larvae were collected from fruit orchards in regions where CpGV-based biopesticides have been extensively used and then discontinued (Catamarca) or are still being used (Alto Valle). The survey was also extended to regions where few occasional (i.e. Mendoza) or no virus treatments (various locations) have been made. PCR analysis showed an unexpectedly high proportion of infected larvae (31.3-50%) at most sampling sites in Catamarca. Phylogenetic analysis of these samples, based on gran and lef-8 genes partial sequences, revealed a previously unreported CpGV variant which could not be assigned to any of the known CpGV genome types and formed a well-defined new cluster. Instead, samples from Alto Valle were genetically similar to isolate CpGV-M (the active ingredient in commercial formulations). There, the percentage of infected individuals was considerably lower (1.8-21.9%). The virus was not detected in Mendoza or the other sampling regions. Implications of these results on the epidemiology and current classification of CpGV isolates are discussed. Fil: Arneodo Larochette, Joel Demián. 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; Argentina Fil: De Anna, J.. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Microbiología y Zoología Agrícola; Argentina Fil: Salvador, R.. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Microbiología y Zoología Agrícola; Argentina Fil: Farinon, M.. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Microbiología y Zoología Agrícola; Argentina Fil: Quintana, G.. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Microbiología y Zoología Agrícola; Argentina Fil: Sciocco Cap, A.. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Microbiología y Zoología Agrícola; Argentina
- Published
- 2015
34. Development of a recombination system for the generation of occlusion positive genetically modified Anticarsia Gemmatalis Multiple Nucleopolyhedrovirus
- Author
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M. Leticia Ferrelli, Santiago Haase, Alicia Sciocco-Cap, Víctor Romanowski, Christina B. McCarthy, and Matías Luis Pidre
- Subjects
lcsh:QR1-502 ,Anticarsia gemmatalis ,Recombinant virus ,lcsh:Microbiology ,law.invention ,purl.org/becyt/ford/1 [https] ,Genes, Reporter ,law ,Polyhedrin ,Recombination, Genetic ,Leguminosas ,Genetically Modified Organisms ,Legumes ,Genetically modified organism ,Velvetbean caterpillar ,Lepidoptera ,Infectious Diseases ,Organismos Modificados Genéticamente ,Plagas de Plantas ,Larva ,velvetbean caterpillar ,Recombinant DNA ,CIENCIAS NATURALES Y EXACTAS ,Biopesticides ,Bioplaguicidas ,Otras Ciencias Biológicas ,Green Fluorescent Proteins ,Heterologous ,Biology ,Article ,Ciencias Biológicas ,Recombinant baculovirus ,Virology ,Animals ,purl.org/becyt/ford/1.6 [https] ,Molecular Biology ,Gene ,Ciencias Exactas ,recombinant baculovirus ,fungi ,AgMNPV ,South America ,biology.organism_classification ,Nucleopolyhedroviruses ,Bioinsecticide ,Pests of Plants ,bioinsecticide ,Homologous recombination ,Entomology - Abstract
Anticarsia gemmatalis is an important pest in legume crops in South America and it has been successfully controlled using Anticarsia gemmatalis Multiple Nucleopolyhedrovirus (AgMNPV) in subtropical climate zones. Nevertheless, in temperate climates its speed of kill is too slow. Taking this into account, genetic modification of AgMNPV could lead to improvements of its biopesticidal properties. Here we report the generation of a two-component system that allows the production of recombinant AgMNPV. This system is based on a parental AgMNPV in which the polyhedrin gene (polh) was replaced by a bacterial β-galactosidase (lacZ) gene flanked by two target sites for the homing endonuclease I-PpoI. Co-transfection of insect cells with linearized (I-PpoI-digested) parental genome and a transfer vector allowed the restitution of polh and the expression of a heterologous gene upon homologous recombination, with a low background of non-recombinant AgMNPV. The system was validated by constructing a recombinant occlusion-positive (polh+) AgMNPV expressing the green fluorescent protein gene (gfp). This recombinant virus infected larvae normally per os and led to the expression of GFP in cell culture as well as in A. gemmatalis larvae. These results demonstrate that the system is an efficient method for the generation of recombinant AgMNPV expressing heterologous genes, which can be used for manifold purposes, including biotechnological and pharmaceutical applications and the production of orally infectious recombinants with improved biopesticidal properties., Facultad de Ciencias Exactas, Instituto de Biotecnologia y Biologia Molecular
- Published
- 2015
35. Sequencing and Characterisation of p74 Gene in Two Isolates of Anticarsia Gemmatalis MNPV
- Author
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Pablo Daniel Ghiringhelli, Mariano Nicolás Belaich, Vanina Andrea Rodríguez, Alicia Sciocco-Cap, Victor Romanowski, Marcela Gabriela Pilloff, and Marcos Fabian Bilen
- Subjects
Genes, Viral ,Transcription, Genetic ,viruses ,Molecular Sequence Data ,Locus (genetics) ,Genome, Viral ,Biology ,Polymerase Chain Reaction ,Genome ,Complete sequence ,Viral Envelope Proteins ,Virology ,Genetics ,Animals ,Amino Acid Sequence ,Cloning, Molecular ,ORFS ,Molecular Biology ,Peptide sequence ,Gene ,Phylogeny ,Base Sequence ,fungi ,DNA replication ,General Medicine ,Amplicon ,Molecular biology ,Nucleopolyhedroviruses ,Lepidoptera ,DNA, Viral - Abstract
P74 is a protein encoded in the genome of baculoviruses, associated with the envelopes of occluded virus. Its presence proved to be essential for per os infection. In first place, in this work we designed two universal primers to amplify a sequence region of the p74 ORF in baculoviruses from different classification groups. Then, by the use of these amplicons we obtained the complete sequence of the p74 locus from two isolates of AgMNPV, 2D (Brazil) and SF (Argentina). In the flanking regions we determined the complete sequence of p10 gene and a portion of p26 gene. Comparing both p74 sequence data (ORFs of 1935 bp) we found fifteen nucleotide changes that result in six amino acid changes. Comparisons of AgMNPV p74s with other baculovirus homologous genes indicate a close relationship with other group I Nucleopolyhedrovirus, in particular CfDEFNPV. These results were based on ORF sequence, amino acid sequence and gene order. The predictive studies about secondary structure and hydrophobic index point at six regions potentially associated to its function or native conformation. Finally, the detection of p74 mRNA after virus DNA replication confirms a late expression pattern.
- Published
- 2006
36. [Untitled]
- Author
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Alicia Sciocco-Cap, M. Alejandra ManzÁn, Mario Enrique Lozano, P. Daniel Ghiringhelli, Víctor Romanowski, and Alejandro D. Parola
- Subjects
Genetics ,Phylogenetic tree ,General Medicine ,Biology ,Genome ,Restriction enzyme ,Restriction map ,Granulovirus ,Virology ,ORFS ,Molecular Biology ,Gene ,Southern blot - Abstract
The bean shoot borer, Epinotia aporema (Lep. Tortricidae), is an economically important pest of legume crops in South America. Recently, a granulovirus (EpapGV) was isolated from E. aporema larvae, and evaluated as a potential biological control agent. In order to generate a restriction map and to investigate the gene organisation of EpapGV genome, DNA isolated from occlusion bodies as well as a set of cloned genomic fragments were analysed using combinations of restriction endonucleases and Southern blot analyses that lead to a first version of the physical map. It was subsequently confirmed and refined by sequencing the termini of the cloned fragments and assessing their contiguity by comparing the sequences with databases to identify putative ORFs spanning neighbour fragments. This was also aided by PCR amplifications with primers that pointed outwards of the cloned viral DNA. The granulin gene was positioned on the physical map, cloned and sequenced. Its 747-nucleotide-long ORF encodes a predicted protein of 29 kDa and the core of the baculovirus very late promoter ATAAG was found 29 nucleotides upstream the initiation codon. In addition, 27 putative ORFs were located on the map and used to explore the genome organisation by GeneParityPlot against the fully sequenced granulovirus genomes. These data, taken together with the phylogenetic tree generated by alignment of the major occlusion proteins, indicate that EpapGV is closely related to CpGV, but has a distinct gene organisation.
- Published
- 2002
37. [Untitled]
- Author
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Alicia Sciocco-Cap, Pablo Daniel Ghiringhelli, Víctor Romanowski, Maria Alejandra Manzan, and Mario Enrique Lozano
- Subjects
chemistry.chemical_classification ,Untranslated region ,Genetics ,Ecdysteroid ,biology ,Phylogenetic tree ,Epinotia ,General Medicine ,biology.organism_classification ,Amino acid ,chemistry.chemical_compound ,Granulovirus ,chemistry ,Virology ,Protein folding ,Molecular Biology ,Gene - Abstract
The ecdysteroid UDP-glycosyltransferase (egt) gene of Epinotia aporema granulovirus (EpapGV) was cloned sequenced and its biological activity was assessed. It encodes a protein of 446 amino acids. Direct evidence that the cloned gene encodes an active EGT protein was obtained by transient expression assays in insect cells. The upstream untranslated region of the egt gene exhibits several consensus early promoter elements. Accordingly, the gene is expressed early upon infection of Epinotia aporema larvae and the EGT activity remains high until later times post infection. Sequence analyses indicate the presence of clusters of amino acid residues conserved among all the baculoviral EGTs, although their relation with proper protein folding, ligand binding and catalytic activity remain to be assessed. Phylogenetic trees consistently cluster the granulovirus EGTs separating them clearly from the nucleopolyhedroviruses.
- Published
- 2002
38. Transcriptomic Survey of the Midgut of Anthonomus grandis (Coleoptera: Curculionidae)
- Author
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Ricardo Salvador, Alicia Sciocco-Cap, Norma Paniego, Dario Principi, Marcelo Facundo Berretta, Paula Fernández, and Esteban Hopp
- Subjects
Otras Biotecnología Agropecuaria ,COTTON PEST ,Cotton ,CONTROL BIOLÓGICO ,purl.org/becyt/ford/1 [https] ,Transcriptome ,Control de Plagas ,MIDGUT EXPRESSED GENE ,Phylogeny ,biology ,purl.org/becyt/ford/4.4 [https] ,midgut expressed gene ,General Medicine ,Coleoptera ,ARN ,Anthonomus ,Plagas de Plantas ,Larva ,CIENCIAS NATURALES Y EXACTAS ,Molecular Sequence Data ,Biotecnología Agropecuaria ,Ciencias Biológicas ,Biología Celular, Microbiología ,Botany ,Animals ,Computer Simulation ,Amino Acid Sequence ,RNA, Messenger ,purl.org/becyt/ford/1.6 [https] ,business.industry ,Research ,GENÓMICA FUNCIONAL ,Pest control ,cotton pest ,Midgut ,biology.organism_classification ,PICUDO ALGODÓN ,Algodón ,454 PYROSEQUENCING ,Pests of Plants ,CIENCIAS AGRÍCOLAS ,Insect Science ,Curculionidae ,RNA ,Weevils ,454 pyrosequencing ,Pest Control ,business ,Digestive System ,Anthonomus grandis ,purl.org/becyt/ford/4 [https] - Abstract
Anthonomus grandis Boheman is a key pest in cotton crops in the New World. Its larval stage develops within the flower bud using it as food and as protection against its predators. This behavior limits the effectiveness of its control using conventional insecticide applications and biocontrol techniques. In spite of its importance, little is known about its genome sequence and, more important, its specific expression in key organs like the midgut. Total mRNA isolated from larval midguts was used for pyrosequencing. Sequence reads were assembled and annotated to generate a unigene data set. In total, 400,000 reads from A. grandis midgut with an average length of 237 bp were assembled and combined into 20,915 contigs. The assembled reads fell into 6,621 genes models. BlastX search using the NCBI-NR database showed that 3,006 unigenes had significant matches to known sequences. Gene Ontology (GO) mapping analysis evidenced that A. grandis is able to transcripts coding for proteins involved in catalytic processing of macromolecules that allows its adaptation to very different feeding source scenarios. Furthermore, transcripts encoding for proteins involved in detoxification mechanisms such as p450 genes, glutathione-S-transferase, and carboxylesterases are also expressed. This is the first report of a transcriptomic study in A. grandis and the largest set of sequence data reported for this species. These data are valuable resources to expand the knowledge of this insect group and could be used in the design of new control strategies based in molecular information. Fil: Salvador, Ricardo. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Microbiología y Zoología Agrícola; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina Fil: Perez Principi, Ricardo Dario. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Berretta, Marcelo Facundo. 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; Argentina Fil: Fernández, Paula del Carmen. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Paniego, Norma Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina Fil: Sciocco Cap, Alicia. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Microbiología y Zoología Agrícola; Argentina Fil: Hopp, Horacio Esteban. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Biotecnología; Argentina
- Published
- 2014
39. Biology and morphometrics of the immature stages of Epinotia aporema on artificial diet
- Author
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Alicia Sciocco-Cap, Joel D. Arneodo, and Graciela Quintana
- Subjects
Glycine max ,Otras Ciencias Biológicas ,Zoology ,larvae ,LARVAE ,Biology ,cápsula cefálica ,Ciencias Biológicas ,purl.org/becyt/ford/1 [https] ,laboratory rearing ,cria em laboratório ,larvas ,purl.org/becyt/ford/1.6 [https] ,Egg incubation ,Morphometrics ,Larva ,GLYCINE MAX [INDEX TERMS] ,Epinotia ,fungi ,biology.organism_classification ,Pupa ,Horticulture ,broca-das-axilas ,Instar ,Animal Science and Zoology ,bean shoot borer ,cephalic capsule ,Agronomy and Crop Science ,CIENCIAS NATURALES Y EXACTAS - Abstract
O objetivo deste trabalho foi estabelecer uma tabela de vida para os estágios imaturos de Epinotia aporema, como parte de um estudo mais amplo para seu controle biológico. Os insetos foram criados em dieta artificial a 25±1ºC e 16:8 (luz:escuridão) horas de fotoperíodo. Para a identificação dos estágios larvais para estudos de interação inseto‑patógeno em condições de laboratório, as larguras de cápsula cefálica também foram determinadas. O período de incubação dos ovos foi de 4,13±0,30 dias, o estágio larval foi de 11,64±0,49 dias, e o tempo de desenvolvimento das pupas dependeu do sexo, com 8,51±0,69 dias para as fêmeas e 9,41±0,65 dias para os machos. Foram identificados cinco estágios larvais. The objective of this work was to establish a life table for the immature stages of Epinotia aporema, as part of a wider investigation on its biological control. Insects were reared on an artificial diet at 25±1ºC and a 16:8 (light:dark) hour photoperiod. For the identification of larval instars for the study of pathogen‑insect interactions under laboratory conditions, head capsule widths (HCWs) were also determined. The egg incubation period was 4.13±0.30 days, larval stage took 11.64±0.49 days, and the development time of the pupal phase was sex‑dependent with 8.51±0.69 days for the females and 9.41±0.65 days for the males. Five larval instars were identified. Fil: Arneodo Larochette, Joel Demián. 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; Argentina Fil: Quintana, Graciela. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Microbiología y Zoología Agrícola; Argentina Fil: Sciocco, Alicia Inés. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Microbiología y Zoología Agrícola; Argentina
- Published
- 2010
40. Analysis of a chitinase from EpapGV, a fast killing betabaculovirus
- Author
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Alicia Sciocco-Cap, Víctor Romanowski, Ricardo Salvador, and M. Leticia Ferrelli
- Subjects
food.ingredient ,KDEL ,Otras Ciencias Biológicas ,Biología ,Sequence alignment ,law.invention ,Ciencias Biológicas ,Open Reading Frames ,food ,law ,Virology ,Betabaculovirus ,Genetics ,Baculovirus ,Molecular Biology ,Phylogeny ,DNA Primers ,EpapGV ,chemistry.chemical_classification ,biology ,Base Sequence ,Virulence ,Chitinases ,Chitinase ,General Medicine ,biology.organism_classification ,V-CHIA ,Molecular biology ,Amino acid ,Open reading frame ,Anticarsia gemmatalis ,chemistry ,biology.protein ,Recombinant DNA ,Peritrophic membrane ,Baculoviridae ,CIENCIAS NATURALES Y EXACTAS - Abstract
The main function of baculoviral chitinase protein (V-CHIA) is to promote the final liquefaction of infected host larvae, facilitating the dispersion of occlusion bodies (OBs) in the environment. In this study, a v-chiA from Epinotia aporema Granulovirus (EpapGV) was identified and characterized. The 1,713 base pairs long open reading frame encodes a protein of 570 amino acids with a predicted molecular weight of 63 kDa. EpapGV V-CHIA sequence alignment resulted 62 % identical to Pieris rapae GV and Blastp search revealed a high conservation among all baculovirus chitinases. Amino acid sequence analysis indicated that the C-terminal KDEL present in most alphabaculovirus chitinases is absent in EpapGV V-CHIA, as well as in the rest of the betabaculoviruses. Phylogenetic analysis was performed with bacterial, lepidopteran, and baculoviral chitinase sequences available in databases. Using an AcMNPV bacmid (bApGOZA) a recombinant Ac-chiAEpapGV was obtained in order to overexpress EpapGV V-CHIA in cell culture. The presence of chitinase was detected in purified AcMNPV-chiAEpapGV OBs. Peritrophic membranes of Anticarsia gemmatalis larvae fed with recombinant OBs showed an altered structure. The results presented in this study show that EpapGV chitinase overexpression in recombinant baculovirus can cause association of this protein with OBs, and suggest that this could be used to evaluate the protein role in early stages of baculoviral infections., Instituto de Biotecnología y Biología Molecular
- Published
- 2013
41. Characterization of a new Helicoverpa armigera nucleopolyhedrovirus variant causing epizootic on a previously unreported host, Helicoverpa gelotopoeon (Lepidoptera: Noctuidae)
- Author
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Ferrelli, M.L., primary, Taibo, C., additional, Fichetti, P., additional, Sciocco-Cap, A., additional, and Arneodo, J.D., additional
- Published
- 2016
- Full Text
- View/download PDF
42. Double-stranded RNA synthesized in bacteria can be transferred to bee and varroa tissues
- Author
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Pablo A. Nakaya, José María Niz, Ricardo Salvador, Angélica Gamboa Cedeño, and Alicia Sciocco-Cap
- Subjects
0106 biological sciences ,0301 basic medicine ,Beekeeping ,fungi ,RNA ,Biology ,biology.organism_classification ,01 natural sciences ,Virology ,010602 entomology ,03 medical and health sciences ,RNA silencing ,030104 developmental biology ,RNA interference ,Transcription (biology) ,Insect Science ,Deformed wing virus ,Varroa destructor ,Varroa - Abstract
In Argentina, the ectoparasite Varroa destructor and deformed wing virus (DWV) causes serious losses in local beekeeping. New control methods have been proposed based on the RNA interference technique. Previous reports showed that bees fed with double-stranded RNA (dsRNA) synthesized in vitro reduce the transcription levels of the target genes in DWV and the varroa mite. An efficient and inexpensive alternative to produce dsRNA is the use of bacteria capable of achieving a high level of in vivo synthesis. In this work, the possibility to use bacteria-mediated delivery of dsRNA to bees and varroa tissues was evaluated. The obtained results indicated that bacterially expressed dsRNA can be transferred to the bee and from bee to the varroa mite.
- Published
- 2015
43. Effect of the interaction between Anticarsia gemmatalis multiple nucleopolyhedrovirus and Epinotia aporema granulovirus, on A. gemmatalis (Lepidoptera: Noctuidae) larvae
- Author
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Biedma, Marina E., primary, Salvador, Ricardo, additional, Ferrelli, M. Leticia, additional, Sciocco-Cap, Alicia, additional, and Romanowski, Víctor, additional
- Published
- 2015
- Full Text
- View/download PDF
44. Functional analysis of Spodoptera frugiperda nucleopolyhedrovirus late expression factors in Sf9 cells
- Author
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Oscar Taboga, M. Gabriela López, Alicia Sciocco-Cap, Víctor Romanowski, and Marcelo Facundo Berretta
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Gene Expression Regulation, Viral ,animal structures ,viruses ,Gene Expression ,Sf9 ,Biology ,Spodoptera ,Genome ,Genes, Reporter ,Virology ,Gene expression ,Late expression factors ,Genetics ,Sf9 Cells ,Animals ,SfMNPV ,Promoter Regions, Genetic ,Molecular Biology ,Gene ,Ciencias Exactas ,Regulation of gene expression ,Functional analysis ,fungi ,General Medicine ,biochemical phenomena, metabolism, and nutrition ,AcMNPV ,biology.organism_classification ,Molecular biology ,Nucleopolyhedroviruses ,Lepidoptera ,Gene cassette ,embryonic structures - Abstract
We used transient expression assays to assess the function of the baculovirus Spodoptera frugiperda M nucleopolyhedrovirus (SfMNPV) homologs of Autographa californica MNPV (AcMNPV) factors involved in late gene expression (lefs), in the Sf9 insect cell-line, which is permissive for both viruses. It is well-established that nineteen AcMNPV lefs support optimal levels of activity from a late promoter-reporter gene cassette in this assay. A subgroup of SfMNPV lefs predicted to function in transcription-specific events substituted the corresponding AcMNPV lefs very efficiently. When all SfMNPV lefs were assayed, including replication lefs, activity was low, but addition of two AcMNPV lefs not encoded in SfMNPV genome, resulted in augmented reporter activity. SfMNPV IE-1 was able to activate an early promoter cis-linked to an hr-derived element from SfMNPV but not from AcMNPV. However, the level of early promoter activation with SfMNPV IE-1 was lower compared to AcMNPV IE-1., Facultad de Ciencias Exactas, Instituto de Biotecnología y Biología Molecular
- Published
- 2013
45. Biological and molecular features of <italic>Nosema rachiplusiae</italic> sp. n., a microsporidium isolated from the neotropical moth <italic>Rachiplusia nu</italic> (Guenée) (Lepidoptera: Noctuidae).
- Author
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Sciocco-Cap, Alicia and Arneodo, Joel D.
- Subjects
- *
NOSEMA , *ULTRASTRUCTURE (Biology) , *HISTOPATHOLOGY , *PHYLOGENY , *DNA analysis - Abstract
Light, electron microscopy and DNA analyses were performed to characterize a microsporidium infecting
Rachiplusia nu larvae from a laboratory rearing in Argentina. Diplokaryotic spores were oval and measured 3.61 ± 0.29 × 1.61 ± 0.14 μM (fresh). The spore wall was composed of an electron-dense exospore and an electron-lucent endospore, ca. 30 nm and 100-120 nm thick, respectively. The polar filament was arranged in a single rank of 10-12 coils (typically 11). Microsporidian cells were found in the cytoplasm, next to the endoplasmic reticulum (especially the prespore stages) and generally surrounded by electron-lucent spaces. The infection was polyorganotropic; the fat body appeared as the most heavily invaded tissue, followed by tracheal matrix and epidermis. A molecular phylogeny based on the small (SSU) and large subunit (LSU) ribosomal RNA genes clearly placed the new isolate within the “Nosema bombycis clade”. Considering both SSU and LSU concatenated partial sequences, the microsporidium fromR. nu showed 99.5% nucleotide similarity withN. bombycis and 99.8% with its closest relative, a microsporidium isolated fromPhilosamia cynthia . According to its genetic and biological features, theR. nu isolate is proposed as the new speciesNosema rachiplusiae sp. n., expanding the limited knowledge on microsporidia associated to endemic South-American moths. [ABSTRACT FROM AUTHOR]- Published
- 2018
- Full Text
- View/download PDF
46. Genome of Epinotia aporema granulovirus (EpapGV), a polyorganotropic fast killing betabaculovirus with a novel thymidylate kinase gene
- Author
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Ricardo Salvador, Pablo Daniel Ghiringhelli, Santiago Haase, Marcelo Facundo Berretta, Marina Elizabeth Biedma, Alicia Sciocco-Cap, Víctor Romanowski, María Leticia Ferrelli, Instituto de Biotecnología y Biología Molecular [La Plata] (IBBM), Consejo Nacional de Investigaciones Científicas y Técnicas [Buenos Aires] (CONICET)-Facultad de Ciencias Exactas [La Plata], Universidad Nacional de la Plata [Argentine] (UNLP)-Universidad Nacional de la Plata [Argentine] (UNLP), Instituto de Microbiología y Zoología Agrícola (IMyZA), Instituto Nacional de Tecnología Agropecuaria (INTA)-Centro de Investigación en Ciencias Veterinarias y Agronómicas (CICVYA), Interaction virus-hôte et maladies du foie, Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratorio de Ingeniería Genética y Biología Celular y Molecular - Area Virosis de Insectos [Buenos Aires] (LIGBCM-AVI), Instituto de Microbiología Básica y Aplicada [Buenos Aires] (IMBA), Departamento de Ciencia y Tecnología [Buenos Aires], Universidad Nacional de Quilmes (UNQ)-Universidad Nacional de Quilmes (UNQ)-Departamento de Ciencia y Tecnología [Buenos Aires], Universidad Nacional de Quilmes (UNQ)-Universidad Nacional de Quilmes (UNQ), This work was supported by grants from the Agencia Nacional de Promoción Científica y Tecnológica (ANPCyT) and UNLP to VR and ASC and the Instituto Nacional de Tecnología Agropecuaria (INTA) to ASC., and BMC, Ed.
- Subjects
food.ingredient ,lcsh:QH426-470 ,Sequence analysis ,lcsh:Biotechnology ,Molecular Sequence Data ,structural gene ,Granulovirus ,Genome, Viral ,Biology ,phylogeny ,Genome ,thymidylate kinase gene ,03 medical and health sciences ,food ,Betabaculovirus ,[SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN] ,lcsh:TP248.13-248.65 ,Genetics ,Animals ,controlled study ,Amino Acid Sequence ,Gene ,Ciencias Exactas ,030304 developmental biology ,Synteny ,0303 health sciences ,030306 microbiology ,Nucleic acid sequence ,nucleotide sequence ,Sequence Analysis, DNA ,3. Good health ,Lepidoptera ,genetic code ,lcsh:Genetics ,GenBank ,DNA, Viral ,[SDV.BBM.GTP] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN] ,Nucleoside-Phosphate Kinase ,Research Article ,Biotechnology - Abstract
Background: Epinotia aporema (Lepidoptera: Tortricidae) is an important pest of legume crops in South America. Epinotia aporema granulovirus (EpapGV) is a baculovirus that causes a polyorganotropic infection in the host larva. Its high pathogenicity and host specificity make EpapGV an excellent candidate to be used as a biological control agent. Results: The genome of Epinotia aporema granulovirus (EpapGV) was sequenced and analyzed. Its circular double-stranded DNA genome is 119,082 bp in length and codes for 133 putative genes. It contains the 31 baculovirus core genes and a set of 19 genes that are GV exclusive. Seventeen ORFs were unique to EpapGV in comparison with other baculoviruses. Of these, 16 found no homologues in GenBank, and one encoded a thymidylate kinase. Analysis of nucleotide sequence repeats revealed the presence of 16 homologous regions (hrs) interspersed throughout the genome. Each hr was characterized by the presence of 1 to 3 clustered imperfect palindromes which are similar to previously described palindromes of tortricid-specific GVs. Also, one of the hrs (hr4) has flanking sequences suggestive of a putative non-hr ori. Interestingly, two more complex hrs were found in opposite loci, dividing the circular dsDNA genome in two halves. Gene synteny maps showed the great colinearity of sequenced GVs, being EpapGV the most dissimilar as it has a 20 kb-long gene block inversion. Phylogenetic study performed with 31 core genes of 58 baculoviral genomes suggests that EpapGV is the baculovirus isolate closest to the putative common ancestor of tortricid specific betabaculoviruses. Conclusions: This study, along with previous characterization of EpapGV infection, is useful for the better understanding of the pathology caused by this virus and its potential utilization as a bioinsecticide., Facultad de Ciencias Exactas
- Published
- 2012
47. Analysis of EpapGV gp37 gene reveals a close relationship between granulovirus and entomopoxvirus
- Author
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M. Leticia Ferrelli, Alicia Sciocco-Cap, Víctor Romanowski, Ricardo Salvador, Marcelo Facundo Berretta, Wataru Mitsuhashi, and Marina Elizabeth Biedma
- Subjects
Glycosylation ,Gene Transfer, Horizontal ,Genes, Viral ,Sequence analysis ,Biología ,Granulovirus ,Biology ,Cydia pomonella granulovirus ,Gene product ,Open Reading Frames ,Viral Proteins ,Viral Envelope Proteins ,Virology ,Genetics ,Animals ,Amino Acid Sequence ,Baculovirus ,Molecular Biology ,Gene ,Phylogeny ,EpapGV ,Base Sequence ,Sequence Homology, Amino Acid ,Immune Sera ,GP37 ,General Medicine ,Lepidoptera ,Open reading frame ,GenBank ,Horizontal gene transfer ,Entomopoxvirinae ,Entomopoxvirus - Abstract
The Epinotia aporema Granulovirus GP37 protein gene has been identified, located, and sequenced. This gene was similar to other baculovirus gp37, to entomopoxvirus fusolin gene, and to the chitin-binding protein gene of bacteria. Sequence analysis indicated that the open reading frame is 669 bp long (the smallest gp37 sequenced at present) and encodes a predicted 222-amino acid protein. This protein is glycosylated and specifically recognized by an entomopoxvirus fusolin antiserum. The pairwise comparison of EpapGV gp37 gene product with all the baculovirus sequences in GenBank yields high similarity values ranging from 45 to 63 % with Cydia pomonella Granulovirus gp37 being the most closely related. The phylogenetic analysis interestingly grouped the granuloviruses in a cluster more closely related to entomopoxviruses than to nucleopolyhedroviruses, suggesting a possible horizontal transfer event between the granulovirus group and the entomopoxvirus group., Instituto de Biotecnología y Biología Molecular
- Published
- 2012
48. The Baculoviral Genome
- Author
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Marcelo Facundo Berretta, Mariano Nicolás Belaich, Alicia Sciocco-Cap, Víctor Romanowski, P. Daniel Ghiringhelli, and M. Leticia Ferrelli
- Subjects
2. Zero hunger ,0303 health sciences ,03 medical and health sciences ,030306 microbiology ,Biology ,Humanities ,Genome ,030304 developmental biology - Abstract
M. Leticia Ferrelli1, Marcelo F. Berretta2, Mariano N. Belaich3, P. Daniel Ghiringhelli3, Alicia Sciocco-Cap2 and Victor Romanowski1 1Instituto de Biotecnologia y Biologia Molecular, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, CONICET 2Laboratorio de Ingenieria Genetica y Biologia Celular y Molecular Area Virosis de Insectos, Departamento de Ciencia y Tecnologia, Universidad Nacional de Quilmes 3Instituto de Microbiologia y Zoologia Agricola, INTA Castelar Argentina
- Published
- 2012
49. Molecular Identification ofHelicoverpa armigera(Lepidoptera: Noctuidae: Heliothinae) in Argentina and Development of a Novel PCR-RFLP Method for its Rapid Differentiation FromH. zeaandH. gelotopoeon
- Author
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Arneodo, Joel D., primary, Balbi, Emilia I., additional, Flores, Fernando M., additional, and Sciocco-Cap, Alicia, additional
- Published
- 2015
- Full Text
- View/download PDF
50. Baculovirus Insecticides in Latin America: Historical Overview, Current Status and Future Perspectives
- Author
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Haase, Santiago, primary, Sciocco-Cap, Alicia, additional, and Romanowski, Víctor, additional
- Published
- 2015
- Full Text
- View/download PDF
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