Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “Spodoptera”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 451 records · Page 25Linked to original sources
PubMed1979

Infection of Spodoptera frugiperda cells with Autographa californica nuclear polyhedrosis virus II. The viral DNA and the kinetics of its replication.

The kinetics of replication of Autographa californica nuclear polyhedrosis virus (AcNPV) DNA in Spodoptera frugiperda cells in culture were studied. Viral DNA replication started at about 5 hr postinfection, the rate of viral DNA replication reached a maximum at about 18 hr postinfection and thereafter decreased. Parental viral DNA could be detected within the first hour postinfection in the total intracellular DNA by the Southern technique. There was no evidence for the occurrence of AcNPV DNA sequences which became covalently linked to cellular DNA between 1 and 3 hr post-infection in the productive cycle. The AcNPV DNA appeared as a covalently closed circular molecule of about 92 x 10(6) daltons. The AcNPV DNA did not seem to be methylated, at least there were no 5'-CmCGG3' sequences detectable in this DNA. Restriction enzyme analysis of viral DNA preparations derived from several single plaque isolates of AcNPV as well as of DNA from different virus stocks revealed a certain heterogeneity of the AcNPV DNA.

Journal Article↗
PubMed1987

Identification and nucleotide sequence of the regions of Autographa californica nuclear polyhedrosis virus genome carrying insertion elements derived from Spodoptera frugiperda.

A morphology mutant of the baculovirus Autographa californica nuclear polyhedrosis virus called M5 was previously shown to synthesize two size classes of viral DNA, one of which had a deletion of 42% of the genome. It was hypothesized that the presence of the smaller M5 circular DNA resulted from the specific deletion of the region located between two sites carrying short DNA insertions. These sites have now been identified by DNA-DNA hybridization using cloned EcoRI fragments of M5 DNA. One cloned M5 EcoRI fragment was found to correspond to the deletion junction fragment where sequences from the insertion site in the 2.6-map unit region were covalently linked to sequences from the insertion site in the 46-map unit region. The 2.6- and 46-map unit regions of Wt AcMNPV DNA were sequenced. Potential long open reading frames which would be disrupted by the M5 inserts were detected. Nucleotide sequence analysis of the same regions of M5 DNA revealed the presence of almost identical inserts of 290 bp. The primary sequence of the inserts revealed characteristics similar to the termini of transposons. Hybridization studies suggested that the inserts were derived from repetitive elements of the Spodoptera frugiperda host cell genome.

Animals↗
PubMed1989

Baculovirus polyhedrin promoter-directed expression of rubella virus envelope glycoproteins, E1 and E2, in Spodoptera frugiperda cells.

To study the capability of Spodoptera frugiperda (fall armyworm; Sf9) cells to synthesize and process mature rubella virus (RV) proteins, a cDNA encoding the structural envelope glycoproteins, E1 (58 kDa) and E2 (42-47 kDa) were inserted into the genome of Autographa californica nuclear polyhedrosis virus (AcNPV) and expressed during infection under the transcriptional regulation of the polyhedrin gene promoter. By immunoblot analysis with antibodies directed against purified RV, the individual proteins E1 and E2, and human convalescent serum, a polyprotein precursor migrating with an apparent molecular weight of 90-95 kDa was identified in Sf9 cells infected with the recombinant baculovirus, Ac701-RVE. In addition, two proteins migrating somewhat faster than authentic viral E1 and E2 were resolved. Pulse-chase labeling experiments in the absence and presence of tunicamycin, as well as treatment of the recombinant proteins with endo-beta-N-acetyl-D-glucosaminidase H indicated that the recombinant proteins are glycosylated and that the E1 and E2 apoproteins, respectively, were similar in size as compared to their in vitro synthesized counterparts. The recombinant protein products were further detected by some monoclonal antibodies directed against RV. The results presented here indicate that a polyprotein containing the envelope glycoproteins of RV is expressed and proteolytically cleaved in lepidopteran insect cells to form two proteins which resemble authentic E1 and E2. The baculovirus may therefore be suitable for the abundant expression of RV antigen.

Animals↗
PubMed1990

Cloning of the genome of a densovirus and rescue of infectious virions from recombinant plasmid in the insect host Spodoptera littoralis.

We have cloned an infectious genome of the Junonia coenia densonucleosis virus (JcDNV) into the bacterial plasmid pBR322. The viral genome could be rescued from the recombinant plasmid pBRJ by transfection of pBRJ DNA to sensitive Spodoptera littoralis larvae. pBRJ DNA produced a typical viral infection and a comparable percentage of larvae became infected following inoculation of equivalent amounts of purified virion DNA or cloned viral DNA. Virions extracted from transfected larvae were indistinguishable from wild-type (wt) virions with regard to their biophysical and biological properties. In particular, rescued virions were as infectious as wt virions and showed identical restriction profiles of their genome. In contrast, subcloning of JcDNV DNA deleted at both extremities of a sequence of ca 250 or ca 100 bp resulted in the inability of the recombinant plasmids to initiate a viral infection. These data suggest that, as for vertebrate parvoviruses, the inverted terminal repeats display essential functions in the rescue process and replicative cycle of densoviruses. This is the first report of the molecular cloning of the infectious genome from an insect parvovirus, and more generally from an invertebrate virus. pBRJ should provide an efficient tool to further define the organization of the JcDNV genome and compare it to other parvoviruses.

Animals↗
PubMed1992

Effects of oxygen on recombinant protein production by suspension cultures of Spodoptera frugiperda (Sf-9) insect cells.

Spodoptera frugiperda (Sf-9) insect cells have been grown in serum-free medium in 250-ml spinner flasks. The maximum cell density obtained in these cultures was dependent on the aeration rate of the culture. Similar yields of uninfected cells were obtained when cultures were stirred in spinner flasks at 80 rev min-1 and in a 4-1 stirred-tank bioreactor and the dissolved oxygen in the bioreactor was controlled at 20% of air saturation. Cells were infected with a recombinant baculovirus at different multiplicities of infection: the timing and maximum level of expression of the recombinant protein were dependent on the multiplicity of infection, the cell density at infection, and on the aeration rate of the culture. Oxygen-limited growth resulted in undetectable levels of recombinant protein (< 6 ng recombinant protein 10(-7) cells). Compared with the maximum yields observed in spinner flask cultures, higher levels of recombinant protein were produced when cells were grown and infected in the bioreactor. The level of dissolved oxygen in the bioreactor was controlled at 50% of air saturation.

Animals↗
PubMed1986

Characteristics of a galactose-binding hemagglutinin (lectin) from hemolymph of Spodoptera exigua larvae.

Hemolymph from Spodoptera exigua larvae agglutinates rabbit and human O erythrocytes. The agglutinin appears to be naturally occurring i.e., injury (by injection) or injection of larvae with fungal cells does not induce an increase in titer of hemolymph samples. Hemagglutinin activity is destroyed by heat or EDTA, and galactosidic carbohydrates inhibit agglutination of both red blood cell types. The agglutinin was purified by affinity chromatography using an Affi-Gel ovalbumin column. Binding to the column is calcium (cation) dependent. SDS gel electrophoresis shows that the agglutinin is a minor component of whole hemolymph represented by two bands with molecular weights of 30,500 and 31,000 daltons. Fluorescence microscopy using rhodamine-labeled agglutinin indicates that the agglutinin binds to fungal cell wall surfaces known to have galactose residues (e.g., Paecilomyces ochraceus), and that binding is specifically inhibited by galactose. There is no specific binding to fungal walls known to lack galactose residues (e.g., Paecilomyces ochraceus, Nomuraea rileyi). The agglutinin may be involved in the immune response of the insect e.g., by opsonization of microbial (fungal) surfaces which render the invading cells more susceptible to phagocytosis or agglutination.

Animals↗
PubMed1990

DNA distribution and respiratory activity of Spodoptera frugiperda populations infected with wild-type and recombinant Autographa californica nuclear polyhedrosis virus.

Spodoptera frugiperda cells were infected with a wild-type Autographa californica nuclear polyhedrosis virus and with a recombinant Autographa californica nuclear polyhedrosis virus. The recombinant virus was derived from the wild-type virus and produced beta-galactosidase instead of polyhedrin. The changes in cell size, cell growth, viability, DNA distribution, and respiratory activity were followed through the time course of the infection. The DNA content as measured by flow cytometry of infected cells increased to approximately 1.8 times the value of uninfected cells and the distributions of single-cell DNA content of the infected cells were strongly deformed. Early in the infection the respiratory activity passed through a maximum. The mitochondrial activity based on Rhodamine 123 labelling of cells infected with the recombinant virus, as determined by flow cytometry, also passed through a maximum at 24 h post infection while the mitochondrial activity of cells infected with the wild-type virus continued to increase. Evolution of single-cell mitochondrial activity was different in uninfected populations and in populations infected with wild-type and with recombinant virus. In all experiments performed, the recombinant virus influenced cell behavior and the measured parameters earlier than the wild-type virus. The influence of the multiplicity of infection was stronger for the wild-type virus than for the recombinant virus.

Animals↗
PubMed1991

Expression and characterization of poliovirus proteins 3BVPg, 3Cpro, and 3Dpol in recombinant baculovirus-infected Spodoptera frugiperda cells.

As an initial step toward investigating the roles of poliovirus proteins in viral RNA replication, a baculovirus expression system was used to produce poliovirus proteins from the P3 region. Spodoptera frugiperda (Sf9) cells were infected with a recombinant baculovirus, vETL-PoV3A*BCD, which contains cDNA coding for poliovirus proteins 3D, 3C, 3B, and a portion of 3A protein sequence. Immunofluorescence microscopy revealed that the majority of 3D (polymerase) was in the cytoplasm of recombinant baculovirus-infected Sf9 cells. In the same cells, the 3C (protease) and 3B (VPg) proteins appeared to be located in distinct subcellular regions, possibly membrane structures, suggesting that the expressed polyprotein was cleaved to generate mature proteins. Processing of the polypeptide was confirmed by immunoblot analysis which demonstrated that 3Cpro sequences were active in cleavage of the polyproteins 3A*BCD and 3CD. Over 95% of the 3D sequences accumulated in the form of mature 3Dpol, with only low levels of 3CD remaining. The majority of 3Dpol remained in the supernatant after low speed centrifugation of sonicated cells. The 3Dpol had RNA-dependent RNA polymerase activity as measured by elongation of an oligo(U) primer using a poly(A) template. The protein 3CDpro was active in cleaving P1 protein. The yield and activities of the poliovirus proteins expressed will facilitate future biochemical studies.

Animals↗
PubMed1991

Structural comparison of the Autographa californica nuclear polyhedrosis virus-induced RNA polymerase and the three nuclear RNA polymerases from the host, Spodoptera frugiperda.

A partial subunit structure has been determined for the novel RNA polymerase that is induced in fall armyworm (Spodoptera frugiperda) cells upon infection with the Autographa californica nuclear polyhedrosis virus (AcNPV). The putative structure includes nine polypeptides; the complexity of this structure is in accord with the high sedimentation coefficient (15S) estimated for this enzyme. A comparison of the putative structure of the virus-induced polymerase with those of the three host nuclear RNA polymerases shows that the structure of the viral polymerase is apparently unlike any of the host nuclear polymerases. This conclusion is reinforced by immunoblot experiments that show no cross-reactivity between the virus-induced polymerase and an antiserum directed against Drosophila RNA polymerase II. The virus-induced RNA polymerase appears at the onset of the late phase of infection and still appears when viral DNA synthesis is blocked by aphidicolin. Thus, the virus-induced polymerase seems to be composed of early viral products.

Animals↗
PubMed1993

Non-homologous recombination between adenovirus and AcNPV DNA fragments in cell-free extracts from insect Spodoptera frugiperda nuclei.

In previous work, we have developed a cell-free system from nuclear extracts of hamster cells to study the mechanism of integrative recombination between adenovirus type 12 (Ad12) DNA and hamster cell DNA (Jessberger et al., 1989; Tatzelt et al., 1992). We have also demonstrated that in insect cells the left terminal fragment of Ad2 DNA can insert by non-homologous recombination into the 32.6 to 34.5 map unit (EcoRI-O) fragment and into other segments of Autographa californica nuclear polyhedrosis virus (AcNPV) DNA (Xiong et al., 1991). We have now imitated this recombination event in vitro by incubating the E1 fragment of Ad2 DNA and the EcoRI-O fragment of AcNPV DNA, both in the plasmid-cloned circular forms, with partly purified nuclear extracts from Spodoptera frugiperda insect cells. Proteins from these extracts have been fractionated by gel filtration. After the reextraction of DNA from the incubation mixture, recombinants generated in this cell-free system have been identified directly with the polymerase chain reaction (PCR) by using Taq polymerase and appropriate primers which are unique to either of the two reaction partners. The recombinants identified are all different. The results of control experiments argue against the possibility that unspecific reaction products might have been generated during PCR. Nucleotide sequence determinations in some of the recombinants localize the sites of genetic exchange between the two partners and assess the non-homologous nature of the reaction. The recombinants are characterized by the presence of short patch homologies at or close to the sites of linkage between the reaction partners, as described earlier in the hamster cell system (Tatzelt et al., 1992). The occurrence of recombinants in the cell-free system can also be demonstrated by a biological test in which potential recombinants are isolated by transfection into recA- strains of Escherichia coli.

Adenoviridae↗
PubMed1989

Cell-free translation in lysates from Spodoptera frugiperda (Lepidoptera: Noctuidae) cells.

1. Conditions for in vitro translation of mRNA in cell-free extracts from cultured Spodoptera frugiperda cells were defined. 2. Incorporation of [35S]methionine into acid-precipitable material increased for approximately 1 hr, and was sensitive to the protein synthesis inhibitors pactamycin and cycloheximide. 3. Micrococcal nuclease-treated lysate, primed with purified rabbit globin mRNA, synthesized a major protein with the size of full length globin, indicating that the lysate supported correct initiation and elongation of polypeptides.

Animals↗
PubMed1992

Isolation and partial characterization of binding proteins for immobilized delta endotoxin from solubilized brush border membrane vesicles of the silkworm, Bombyx mori, and the common cutworm, Spodoptera litura.

1. Brush border membrane vesicles (BBMV) were prepared from the Bombyx mori, and Spodoptera litura, midguts. BBMV was solubilized. 2. Activated delta endotoxin from Bacillus thuringiensis were immobilized. 3. Solubilized BBMV proteins were applied to the toxin column and the proteins bound were analyzed by SDS-PAGE. 4. In the case of B. mori M(r) = 220,000, 150,000 and 130,000 and in the case of S. litura 160,000 bands were detected. 5. The bindings were inhibited by N-acetyl galactosamine and mannose. 6. The binding proteins applied onto a Con A column and eluted by 0.1 M methyl-alpha-glucose, suggesting that hybrid type sugar sidechain may involve in the interaction.

Animals↗
PubMed1993

The galactose binding lectin from the beet armyworm, Spodoptera exigua: distribution and site of synthesis.

The Spodoptera exigua galactose binding lectin, extracted from hemolymph by affinity chromatography, was comprised of large molecular weight aggregates (100-700 kDa). SDS-PAGE of this lectin preparation revealed it to contain 2 subunits of 33.2 and 34.4 kDa in equimolar concentrations. These subunits had similar amino acid profiles, possessed identical N-terminal sequences and reacted equally to a bank of antilectin monoclonal antibodies. By staining Western blots with various lectin conjugate probes, we demonstrated that the 34.4 kDa subunit contains complex mannose residues, suggesting that this subunit is the glycosylated form of the 33.2 kDa subunit. The N-terminal sequence of the S. exigua lectin was distinct from other invertebrate galactose binding lectins. Light microscopy in combination with immunoelectron microscopy was used to localize the S. exigua lectin in the granules of the granulocyte class of hemocytes. Degranulation of these cells resulted in the release of the lectin. Isotope incorporation studies followed by immunoprecipitation with a S. exigua monoclonal antibody demonstrated that the fat body was the major site of lectin synthesis. Similar studies with hemocyte monolayers did not result in the production of detectable levels of 35S-labeled S. exigua lectin.

Amino Acid Sequence↗
PubMed1994

Characterization of the Spodoptera frugiperda TATA-binding protein: nucleotide sequence and response to baculovirus infection.

A cDNA clone containing a 921 bp open-reading frame (307 amino acids; 34 kDa) homologous to the TATA-binding protein (TBP) was isolated and sequenced from a Spodoptera frugiperda cell line that is commonly used in the baculovirus expression system. Analysis of the S. frugiperda TBP (SfTBP) sequence showed that the amino-terminal portion of SfTBP diverged significantly from that of other TBP sequences including Drosophila melanogaster whereas the carboxy-terminal sequence was highly conserved. Southern blot analysis indicated that SfTBP was encoded by a single gene in the S. frugiperda genome. Northern blot analysis indicated that steady-state levels of the 1.3 kb SfTBP transcript declined by 24 h post-infection corresponding to the time of virus-induced inhibition of host-cell transcription. Corresponding western blot analysis showed that TBP protein levels remain constant up to 72 h post-infection.

Amino Acid Sequence↗
PubMed2006

Functional analysis of the allatostatin-A type gene in the cricket Gryllus bimaculatus and the armyworm Spodoptera frugiperda.

Double-stranded RNA (dsRNA) gene interference is an efficient method to silence gene expression in a sequence specific manner. Here we show, that dsRNA targeting the allatostatin (AS)-A type (FGL/I/V-amide) gene of Gryllus bimaculatus (Ensifera, Gryllidae) and Spodoptera frugiperda (Lepidoptera, Noctuidae) injected into freshly moulted larvae or adult crickets and moths produced a rapid and long-lasting reduction in the mRNA levels in various tissues. The effect lasted up to 7 days. Following dsRNA injection, the juvenile hormone (JH) titers in the hemolymph were clearly raised in both species. AS-dsRNA injection induced a reduced body weight in larval and adult crickets and the imaginal moult was incomplete. Silencing allatostatin type-A expression also reduced the egg and testes development in crickets, and the oviposition rate was drastically diminished in both species.

Animals↗
PubMed2000

Possible bases of pseudoparasitism in Spodoptera littoralis larvae stung by Microplitis rufiventris.

The effects of host age and parasitoid female age on the occurrence of 'Pseudoparasitism', using the Spodoptera littoralis-Microplitis rufiventris host-parasitoid system were investigated. The first four larval instars of the host are not equally suitable for parasitoid development. The proportion of pseudoparasitized hosts significantly increases when: (1) the age of the female parasitoid increases; (2) oviposition occurs mostly in fourth instar larvae; (3) a later age of the host instar is used; (4) the mandibles of the newly hatched parasitoid larvae mistakenly attack host interior organs (e.g. Malpighian tubules); and (5) an imperfect growth pattern of teratocytes occurs. The reluctance of female wasps to parasitize fourth instar host larvae is not due to the thickness of host cuticle but possibly due to the unfavourable physiological state of the host larvae. The age of host larvae at the time of parasitization may influence the adverse effects of parasitoid factors (e.g. polydnavirus, venom and teratocytes) on the growth of host larvae. It is suggested that females of M. rufiventris are able to determine the suitability of a potential host instar for the development of their offspring. The cell diameter of M. rufiventris teratocytes increases with increasing age of host larvae at the time of oviposition. The association within the host of living parasitoid larvae and functional teratocytes may be important for the survival of each other and consequently for successful parasitism.

Journal Article↗
PubMed2000

Eicosanoids rescue Spodoptera exigua infected with Xenorhabdus nematophilus, the symbiotic bacteria to the entomopathogenic nematode Steinernema carpocapsae.

Xenorhabdus nematophilus is a pathogenic bacterium causing insect haemolymph septicemia, which leads to host insect death. To address the fundamental mechanisms underlying this haemolymph septicemia, or the immunodepressive response of the host insects following bacterial infection, we tested a hypothesis that the insect immune-mediating eicosanoid pathway is blocked by inhibitory action of the bacterium. Haemocoelic injection of the bacteria into the fifth instar larvae of Spodoptera exigua reduced the total number of living haemocytes with postinjection time and resulted in host death in 16 h at 25 degrees C. The lethal efficacy, described by the median lethal bacterial dose (LD(50)), was estimated as 33 colony-forming units per fifth instar larva of S. exigua. The lethal effect of the bacteria on the infected larvae decreased significantly with the addition of exogenous arachidonic acid (10 µg), a precursor of eicosanoids. In comparison, injections of dexamethasone (10 µg), a specific inhibitor of phospholipase A(2), and other eicosanoid biosynthesis inhibitors elevated significantly the bacterial pathogenicity. Live X. nematophilus induced the infected larvae to form less nodules than did the heat-killed bacteria, but the addition of arachidonic acid increased the number of nodules formed significantly in response to live bacterial injection. The treatment with dexamethasone and other inhibitors, however, decreased the nodule formation after injection of heat-killed bacteria. These results indicate that eicosanoids play a role in the immune response of S. exigua, and suggest strongly that X. nematophilus inhibits its eicosanoid pathway, which then results in immunodepressive haemolymph septicemia.

Journal Article↗