anticarsia gemmatalis nuclear polyhedrosis virus replication in serum-free and serum-reduced insect cell order to develop a financially feasible process to produce anticarsia gemmatalis nuclear polyhedrosis virus in cell culture, we developed a lipidic supplement to replace fetal calf serum in insect cell culture media. the supplement, prepared with an extract of lipids from hen egg yolk, allowed us to reduce the contents of serum in the culture medium from 10% to 1%. iplb-sf-21 cells could be kept along consecutive passages in serum-reduced medium. the replication of agnpv in heyle-supplemented ...19937764195
characterization of the p10 gene region of anticarsia gemmatalis nucleopolyhedrovirus.the anticarsia gemmatalis nucleopolyhedrovirus (agmnpv) p10 gene region was cloned, sequenced and the putative p10 gene expression characterized by northern-blot analysis. sequence analysis of the p10 gene region indicated the presence of two complete open reading frames (orfs) of 713 and 281 nucleotides, which codes for polypeptides of 273 and 93 amino acids, with homology to the p26 and p10 proteins of baculoviruses, respectively. two additional partial orfs, coding for partial polypeptides of ...200212086145
genome of the most widely used viral biopesticide: anticarsia gemmatalis multiple nucleopolyhedrovirus.the genome of anticarsia gemmatalis multiple nucleopolyhedrovirus isolate 2d (agmnpv-2d), which is the most extensively used virus pesticide in the world, was completely sequenced and shown to have 132 239 bp (g+c content 44.5 mol%) and to be capable of encoding 152 non-overlapping open reading frames (orfs). three orfs were unique to agmnpv-2d, one of which (ag31) had similarity to eukaryotic poly(adp-ribose) polymerases. the lack of chia and v-cath may explain some of the success and growth of ...200617030857
nucleotide sequence and phylogenetic analyses of the dna polymerase gene of anticarsia gemmatalis nucleopolyhedrovirus.the dna polymerase from anticarsia gemmatalis nucleopolyhedrovirus (agmnpv) was identified and sequenced, and its amino acid sequence was compared with other viral dna polymerases to identify conserved regions and to reconstruct a phylogenetic tree. the sequence analysis of the agmnpv dna polymerase gene revealed the presence of a 2976 nucleotides open reading frame (orf) encoding a polypeptide of 991 amino acid residues with a predicted molecular mass of 114.7 kda. among the baculovirus dna pol ...200515845260
pathology of anticarsia gemmatalis larvae infected by two recombinant a. gemmatalis multicapsid nucleopolyhedroviruses.light and stereomicroscopy examinations of anticarsia gemmatalis multicapsid nucleopolyhedrovirus (agmnpv)-infected insects were performed in order to follow infection in its host, a. gemmatalis. fourth-instar a. gemmatalis larvae were infected by administration of occluded virus (polyhedra) from two recombinant agmnpv viruses (vagegtdelta-lacz or vaggala2) directly into the larvae foregut. the recombinant virus vagegtdelta-lacz has the beta-galactosidase gene (lac-z) of escherichia coli under t ...200415748993
the gp64 locus of anticarsia gemmatalis multicapsid nucleopolyhedrovirus contains a 3' repair exonuclease homologue and lacks v-cath and chia genes.anticarsia gemmatalis multicapsid nucleopolyhedrovirus (agmnpv) is one of the most successful biological insecticides. in this study, we cloned and sequenced a 12.5 kbp bamhi-d restriction endonuclease fragment of the agmnpv isolate 2d genome that includes the gp64 gene. we compared this highly conserved region with that of other baculoviruses. agmnpv contained two genes, p22.2 and v-trex, in common with choristoneura fumiferana mnpv (cfmnpv) that were not present in other baculoviruses. the v-t ...200414718636
molecular cloning and sequence analysis of the anticarsia gemmatalis multicapsid nuclear polyhedrosis virus gp64 glycoprotein.the gp64 locus of anticarsia gemmatalis multicapsid nucleopolyhedrovirus isolate santa fe (agmnpv-sf) was characterised molecularly in our laboratory. to this end, we have located and cloned a agmnpv-sf genomic dna fragment containing the gp64 gene and sequenced the complete gp64 locus. nucleotide sequence analysis indicated that the agmnpv gp64 gene consists of a 1500 nucleotide open reading frame (orf), encoding a protein of 499 amino acids. of the seven gp64 homologues identified to date, the ...200312680694
enhancement of sf9 cells and baculovirus production employing grace's medium supplemented with milk whey ultrafiltrate.animal cells can be cultured both in basal media supplemented with fetal bovine serum (fbs) and in serum-free media. in this work, the supplementation of grace's medium with a set of nutrients to reduce fbs requirements in spodoptera frugiperda (sf9) cell culture was evaluated, aiming the production of anticarsia gemmatalis nucleopolyhedrovirus (agmnpv) at a cost lower than those for the production using sf900 ii medium. in grace's medium supplemented with glucose, pluronic f68 (pf68) and yeast ...200519003058
production of viral progeny in insect cells undergoing apoptosis induced by a mutant anticarsia gemmatalis nucleopolyhedrovirus.the anticarsia gemmatalis nucleopolyhedrovirus (agmnpv) is the most successful viral biopesticide in use worldwide. we have demonstrated that despite widespread apoptosis and no protein synthesis at 48 h p.i., ufl-ag-286 cells infected with a mutant of agmnpv (vapag), produced significant amounts of budded virus (bvs) and viral dna late in infection. however, a different susceptible cell line (bti-tn5b 1-4) showed no signs of apoptosis and produced 3.5 times more budded virus when infected with ...200111770855
construction of a recombinant anticarsia gemmatalis nucleopolyhedrovirus (agmnpv-2d) harbouring the beta-galactosidase gene.we have constructed a transfer vector (paggal) containing the beta-galactosidase gene under control of the escherichia coli gpt and agmnpv polyhedrin (polh) promoters. the transfer vector was cotransfected with wild type anticarsia gemmatalis nucleopolyhedrovirus (agmnpv) dna into a. gemmatalis (ufl-ag-286) cells and a recombinant baculovirus (vaggala2) was isolated. the beta-galactosidase gene insertion was checked by polymerase chain reaction (pcr) using dna from agmnpv and vaggala2 and primer ...200111556711
heteroflocculation of amidine polystyrene latex and anticarsia gemmatalis nucleopolyhedrovirus as a model system for studying sunlight protection.anticarsia gemmatalis nucleopolyhedrovirus (agmnpv) is a baculovirus specific for the control of an important soybean defoliator. the baculovirus is comprised of double-stranded dna, occluded in a proteinaceous structure called a polyhedron. ultraviolet sunlight is the most destructive factor that affects the persistence of the virus in the field. in the present study, we use a model system wherein the pathogen is covered by another particle of opposite charge in order to test the effectiveness ...200011254268
methods for detection of anticarsia gemmatalis nucleopolyhedrovirus dna in soil.two methods, phenol-ether and magnetic capture-hybridization (mch), were developed and compared with regard to their sensitivities and abilities to extract the dna of the insect baculovirus anticarsia gemmatalis nucleopolyhedrovirus (agmnpv) from soil and to produce dna amplifiable by pcr. laboratory experiments were performed with 0. 25 g of autoclaved soil inoculated with different viral concentrations to optimize both methods of baculovirus dna extraction and to determine their sensitivities. ...199910347006
structural and ultrastructural changes during the infection of ufl-ag-286 cells with the baculovirus agmnpvduring infection of the permissive insect cell line ufl-ag-286 by the baculovirus anticarsia gemmatalis nucleopolyhedrovirus (agmnpv-2d) several morphological changes occur. by 12 h postinfection (h p.i.), the infected cells became round and exhibited a decrease in the number of cytoplasmic projections. by 24 h p.i., it was possible to detect a virogenic stroma inside the cell nucleus, and after 48 h p.i., polyhedral inclusion bodies were observed. some of these morphological modifications are p ...19989784346
in vitro and in vivo host range of anticarsia gemmatalis multiple nuclear polyhedrosis virus.a clone of the wild type (wt) anticarsia gemmatalis multiple nuclear polyhedrosis virus agmnpv, derived from a geographical isolate (hondrina, brazil) and designated agmnpv-cl4-3a1, was used to determine the host range of this virus in six established lepidopteran cell lines: anticarsia gemmatalis (bcirl-ag-am1), helicoverpa zea (bcirl-hz-am1), heliothis virescens (bcirl-hv-am1), helicoverpa armigera (bcirl-ha-am1), trichoplusia ni (tn-cl1), bombyx mori (bmn), and a coleopteran cell line anthono ...19989542640
cell culture derived agmnpv bioinsecticide: biological constraints and bioprocess issues.we have studied parameters for optimizing the spodoptera frugiperda (sf9) cell culture and viral infection for the production of anticarsia gemmatalis multiple nucleopolyhedrosis virus (agmnpv) polyhedra inclusion bodies (pibs) in shaker-schott or spinner bottles and bioreactors. we have assayed the k(l)a of the systems, initial cell seeding, cell culture volume, dissolved oxygen (do), multiplicity of infection (moi), nutrients consumption, and metabolites production. the medium surface oxygen t ...200519003030
the anticarsia gemmatalis nuclear polyhedrosis virus polyhedrin gene region: sequence analysis, gene product and structural comparisons.the genomic region of the anticarsia gemmatalis multiple nucleocapsid nuclear polyhedrosis virus (agmnpv) strain 2d encoding the polyhedrin gene was cloned and mapped, and a 2085 bp sphi-psti fragment containing the gene was sequenced. the polyhedrin polypeptide of the parental isolate agmnpv was manually sequenced, and the amino acid sequence obtained agreed with that deduced from the dna coding region sequence. agmnpv and orgyia pseudotsugata mnpv (opmnpv) are similar in terms of promoter stru ...19921588315
sequencing and characterisation of p74 gene in two isolates of anticarsia gemmatalis mnpv.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 ...200616525736
a morphometric study of the midgut in resistant and non-resistant anticarsia gemmatalis (hübner) (lepidoptera: noctuidae) larvae to its nucleopolyhedrovirus (agmnpv).in this investigation, the anterior and posterior regions of the midgut of resistant (rl) and non-resistant (sl) anticarsia gemmatalis larvae were analyzed morphometrically to characterize different regions along their length. also, this investigation compares the results between sl and rl to improve the understanding of the resistance mechanisms to the virus. histological sections were analyzed in a computerized system and the data were statistically analyzed by the kruskal-wallis test and by m ...200919275904
the inhibitor of apoptosis gene (iap-3) of anticarsia gemmatalis multicapsid nucleopolyhedrovirus (agmnpv) encodes a functional iap.programmed cell death or apoptosis is one of the defense mechanisms used by insect cells in response to baculovirus infection. baculoviruses harbour antiapoptotic genes to prevent apoptosis and to maintain the normal course of infection. in this work, we showed that, like other baculoviruses, anticarsia gemmatalis multicapsid nucleopolyhedrovirus (agmnpv) has a functional inhibitor of apoptosis gene (iap-3). the iap-3 gene was cloned, sequenced and its transcription confirmed by rt-pcr. the puta ...200515834654
anticarsia gemmatalis multicapsid nucleopolyhedrovirus v-trex gene encodes a functional 3' to 5' exonuclease.the viral three-prime repair exonuclease (v-trex) gene of the anticarsia gemmatalis multicapsid nucleopolyhedrovirus (agmnpv) is the first baculovirus gene to be described with significant homology to a 3' exonuclease. v-trex is an early gene that is expressed by agmnpv from 3 h post-infection. in the present study, the agmnpv v-trex orf was cloned into the baculovirus autographa californica multicapsid nucleopolyhedrovirus (acmnpv) under the control of a polyhedrin promoter. the resulting virus ...200415448348
production of the anticarsia gemmatalis multiple nucleopolyhedrovirus in serum-free suspension cultures of the saufl-ag-286 cell line in stirred reactor and airlift reactor.the velvetbean caterpillar, anticarsia gemmatalis hübner (lepidoptera: noctuidae), is one of the main plagues for soybean crops. velvetbean caterpillar larvae are susceptible to be infected by occlusion bodies of the baculovirus anticarsia gemmatalis multiple nucleopolyhedrovirus (agmnpv), a biological insecticide. the insect cell line saufl-ag-286 produces very high yields of occlusion bodies of agmnpv in suspension cultures done in the low-cost serum-free medium unl-10 in shake-flasks. however ...201121906626
production of selected baculoviruses in newly established lepidopteran cell key to the in vitro mass production of baculoviruses is the development of insect cell lines capable of producing high levels of extracellular virus (ecv) and/or occlusion bodies (obs). for this study, 34 newly established cell lines from 10 lepidopteran species were screened for their ability to produce ecv and obs from a variety of baculoviruses. the selected baculoviruses included: the alfalfa looper virus (acmnpv); the celery looper virus (afmnpv); the velvetbean caterpillar virus (agmnp ...200111515971
physical maps and virulence of anticarsia gemmatalis nucleopolyhedrovirus genomic variants.seventeen plaque purified isolates of two viral preparations of anticarsia gemmatalis multiple nucleopolyhedrovirus (agmnpv), were analyzed in terms of the genomic changes after digestion of their dnas with hindiii and psti restriction enzymes. the 1979 agmnpv wild type preparation (agmnpv-'79) resulted in six different variants and the 1985 viral commercial preparation (agmnpv-'85), in eleven. the genomic variation of all the isolates was mapped showing that those from 1985 presented more heter ...199910550671
replication of anticarsia gemmatalis nuclear polyhedrosis virus in four lepidopteran cell lines.anticarsia gemmatalis nuclear polyhedrosis virus (agnpv; family baculoviridae) is pathogenic for larvae of the velvetbean caterpillar, anticarsia gemmatalis hübner-an important pest of soybean. agnpv is a viable alternative to chemical control of a. gemmatalis in brazil, where its use as a pesticide has brought significant economic and environmental benefits. although a significant amount of information is available on the ecological and biological control aspects of agnpv, very little is known ...19979028926
a variable region of anticarsia gemmatalis nuclear polyhedrosis virus contains tandemly repeated dna sequences.a variable region (psti-t fragment) of two genotypic variants of the anticarsia gemmatalis multiple nucleocapsid nuclear polyhedrosis virus (agmnpv) was sequenced and compared. this region, which is known to have deletions and duplications in agmnpv variants, was shown to be formed of units of a 127 bp tandemly repeated sequence containing two 30 bp imperfect palindromes. southern blot experiments showed that the psti-t fragment contained one of the four homologous regions mapped interspersed in ...19968738171
accumulation of few-polyhedra mutants upon serial passage of anticarsia gemmatalis multiple nucleopolyhedrovirus in cell culture.anticarsia gemmatalis nucleopolyhedrovirus (agmnpv) has been widely used to control the velvetbean caterpillar, anticarsia gemmatalis, in brazil. to date, agmnpv has been produced by larval infection and, due to in vivo production limitations and the continuing high demand for the biopesticide, attempts should be made to develop in vitro production of this virus. in order to investigate the effects caused by serial passage of agmnpv in cell culture, we carried out a total of ten passages and ana ...200919135449
molecular analysis of a mutant anticarsia gemmatalis multiple nucleopolyhedrovirus (agmnpv) shows an interruption of an inhibitor of apoptosis gene (iap-3) by a new class-ii piggybac-related insect transposon.a new piggybac-related transposable element (te) was found in the genome of a mutant anticarsia gemmatalis multiple nucleopolyhedrovirus interrupting an inhibitor of apoptosis gene. this mutant virus induces apoptosis upon infection of an anticarsia gemmatalis cell line, but not in a trichoplusia ni cell line. the sequence of the new te (which was named idt for iap disruptor transposon) has 2531 bp with two dna sequences flanking a putative transposase (tpase) orf of 1719 bp coding for a protein ...200919788700
characterization of the ecdysteroid udp-glucosyltransferase (egt) gene of anticarsia gemmatalis nucleopolyhedrovirus.the anticarsia gemmatalis nucelopolyhedrovirus (agmnpv) egt gene was cloned, sequenced and its expression characterized by rt-pcr and western blot analysis. sequence analysis of the gene indicated the presence of an open reading frame (orf) of 1482 nucleotides, which codes for a polypeptide of 494 amino acids. atata box and a conserved regulatory sequence (catt) found in other baculovirus early genes were present in the promoter region of the egt gene. a poly-a consensus sequence was present in ...200111210933
host-specific transcription of nucleopolyhedrovirus gene homologues in productive and abortive anticarsia gemmatalis mnpv a previous report, we showed that anticarsia gemmatalis nucleopolyhedrovirus (agmnpv) infections of choristoneura fumiferana ipri-cf-124t and bombyx mori bm-5 cell lines are abortive, whereas a. gemmatalis ufl-ag-286 cells efficiently produce infectious virus and polyhedral inclusion bodies (pibs). in the present study, we explored transcription patterns in these infections using representative temporal classes of autographa californica mnpv (acmnpv) genes. northern analyses were carried out ...199910446647
morphological characterization of anticarsia gemmatalis m nucleopolyhedrovirus infection in haemocytes from its natural larval host, the velvet bean caterpillar anticarsia gemmatalis (hübner) (lepidoptera: noctuidae).for a better understanding of virus x host interactions, transmission electron microscopy was used to characterize the intrahaemocoelic infection of anticarsia gemmatalis larval haemocytes by a. gemmatalis m nucleopolyhedrovirus (agmnpv). at 12 h post-infection (h p.i.), we observed nuclear hypertrophy, budded virus assembling, and protrusion towards the cytoplasm, virion envelopment, and accumulation of fibrillar aggregates in the cytoplasm. around 24 h p.i., fibrillar aggregates also appeared ...200415140594
interactions among insect-resistant soybean genotypes extracts with populations of anticarsia gemmatalis hübner (lepidoptera: noctuidae) susceptible and resistant to its nucleopolyhedrovirus.anticarsia gemmatalis nucleopolyhedrovirus (agmnpv) is being used in brazil as a biological insecticide. host plant resistance of soybean to insects is been searched for and some authors have mentioned the interference of plant chemistry in virus efficiency. interactions among soybean extracts of genotypes used as a source of resistance (pi 274454 and pi 227687) with different agmnpv concentrations in populations of a. geatalis susceptible (s) and resistant (r) to the virus were studied at labor ...200919893908
isolation and characterization of a baculovirus associated with the insect parasitoid wasp, cotesia marginiventris, or its host, trichoplusia ni.a multiple nucleopolyhedrovirus (mnpv) was isolated from trichoplusia ni (hübner) (lepidoptera: noctuidae) larvae that had been stung by the parasitoid cotesia marginiventris (cresson) (hymenoptera: braconidae). the wild type virus was plaque purified by infecting a heliothis subflexa (bcirl- hsam1) cell line and isolating several clones. the mean estimated genomic size of this virus based on psti, bsteii, styi, hindiii restriction profiles was estimated to be 106 +/- 2.5 kbp (mean+/-se). a clon ...200820334593
the role of peritrophic membrane in the resistance of anticarsia gemmatalis larvae (lepidoptera: noctuidae) during the infection by its nucleopolyhedrovirus (agmnpv).the aim of this study was to analyze morphologically the peritrophic membrane (pm) of anticarsia gemmatalis larvae resistant (rl) and non-resistant (susceptible) (sl) to the a. gemmatalis multicapsid nucleopolyhedrovirus (agmnpv), in the presence of viral infection. also, in this investigation the results between sl and rl were compared to improve the understanding of the resistance mechanisms to the virus. the pm of sl of a. gemmatalis was less efficient as a barrier against the viral infection ...201121775204
heteroflocculation of sulfate polystyrene latex and anticarsia gemmatalis nucleopolyhedrovirus as a model system for studying sunlight protection.anticarsia gemmatalis nucleopolyhedrovirus (agmnpv) is a baculovirus widely used as a pest control agent. data on field persistence of this microbial insecticide reveal that sunlight is the most destructive of the environmental factors. the introduction of a physical barrier to protect the baculovirus from sunlight has been proposed. in the present work we report on the coating of a baculovirus surface with a uniform layer of smaller particles. the coating was carried out at ph 3, a condition wh ...200111426996
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