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Sample records for por spodoptera frugiperda

  1. RNAi Screening in Spodoptera frugiperda.

    PubMed

    Ghosh, Subhanita; Singh, Gatikrushna; Sachdev, Bindiya; Kumar, Ajit; Malhotra, Pawan; Mukherjee, Sunil K; Bhatnagar, Raj K

    2016-01-01

    RNA interference is a potent and precise reverse genetic approach to carryout large-scale functional genomic studies in a given organism. During the past decade, RNAi has also emerged as an important investigative tool to understand the process of viral pathogenesis. Our laboratory has successfully generated transgenic reporter and RNAi sensor line of Spodoptera frugiperda (Sf21) cells and developed a reversal of silencing assay via siRNA or shRNA guided screening to investigate RNAi factors or viral pathogenic factors with extraordinary fidelity. Here we describe empirical approaches and conceptual understanding to execute successful RNAi screening in Spodoptera frugiperda 21-cell line. PMID:27581295

  2. Effects of heme precursors on CYP1A2 and POR expression in the baculovirus/Spodoptera frugiperda system☆

    PubMed Central

    Lu, Huiyuan; Ma, Jun; Liu, Nian; Wang, Shoulin

    2010-01-01

    Objective CYP1A2 and NADPH-CYP450 oxidoreductase (POR) were expressed in the baculovirus/Spodoptera frugiperda (sf9) system. The aim of this study was to investigate the effects of heme precursors on the expression of CYP1A2 and POR. Methods The heme precursors [δ-Aminolaevulinic Acid (5-ALA), Fe3+ and hemin] were introduced into the system to evaluate their effects on the expression of CYP1A2, POR and their co-expression. All the proteins were identified using immunoblotting, CO-difference spectroscopy, or cytochrome c assay. Results In the present study, functional CYP1A2 and POR were successfully expressed in the baculovirus/sf9 system, and both of them showed high activities. Co-addition of 5-ALA and Fe3+ significantly improved expression of CYP1A2 by about 50% compared with the addition of 5-ALA, Fe3+ or hemin alone. Either co-addition of 5-ALA and Fe3+ or addition of 5-ALA or Fe3+ alone improved the POR expression level 2 fold and its activity 7-10 fold compared with control (no addition). However, unlike CYP1A2, there was no difference between the co-addition and addition of these heme precursors alone. Different ratios of BvCYP1A2 to BvPOR also affected the co-expression of CYP1A2 and POR, with a 3:1 ratio of BvCYP1A2 / BvPOR significantly increasing their co-expression. Surprisingly, the addition of 0.1 mM 5-ALA or Fe3+ alone, but not their co-addition, could significantly improve the CYP1A2 and POR co-expression (P < 0.05). Conclusion 5-ALA and Fe3+ increased the expression of CYP1A2 and POR in a baculovirus/sf9 system, but the pattern of their expression was different between their expression alone and co-expression. PMID:23554636

  3. Expression and purification of recombinant nattokinase in Spodoptera frugiperda cells.

    PubMed

    Li, Xiaoxiang; Wang, Xiaoli; Xiong, Shaoling; Zhang, Jing; Cai, Litao; Yang, Yanyan

    2007-10-01

    A recombinant baculovirus, rv-egfp-NK, containing a reporter gene encoding the enhanced green fluorescent protein (EGFP), was used to express nattokinase (NK), a fibrinolytic enzyme, in Spodoptera frugiperda (SF-9) cells. The recombinant protein also included a histidine tag for purification using Ni(2+) resins. The recombinant NK, approximately 30 kDa, retained fibrinolytic activity (60 U/ml). The integration of the EGFP expression cassette in the Bac-to-Bac system is thus an effective method for the expression and purification of recombinant NK protein in Spodoptera frugiperda insect cells. PMID:17581705

  4. Fall armyworm (Spodoptera frugiperda) migration pathways in the United States

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Spodoptera frugiperda (J.E. Smith; Lepidoptera: Noctuidae) or fall armyworm is an important agricultural pest of a number of crops in the western hemisphere. Two morphologically identical host strains of fall armyworm exist, the rice-strain and corn-strain, with the latter inflicting substantial ec...

  5. Pheromonal divergence between two strains of Spodoptera frugiperda

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Abstract- Spodoptera frugiperda consists of two genetically and behaviorally different strains, the corn- and the rice-strain, which seem to be in the process of sympatric speciation. We investigated the role of strain-specific sexual communication as a prezygotic mating barrier between both strains...

  6. Parasitoids of Fall Armyworm, Spodoptera frugiperda, in Florida

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fall armyworm, Spodoptera frugiperda, attacks a wide variety of host plants and is a serious pest of several crops in Florida. Although no one species is a major biological control agent, FAW does have a suite of parasitoids that may affect populations. These species include the braconids Chelonu...

  7. Development and validation of real-time PCR tests for the identification of four Spodoptera species: Spodoptera eridania, Spodoptera frugiperda, Spodoptera littoralis, and Spodoptera litura (Lepidoptera: Noctuidae).

    PubMed

    Van de Vossenberg, B T L H; Van der Straten, M J

    2014-08-01

    The genus Spodoptera comprises 31 species, 4 of which are listed as quarantine pests for the European Union: Spodoptera eridania (Cramer), Spodoptera frugiperda (Smith), Spodoptera littoralis (Boisduval), and Spodoptera litura (F.). In international trade, the earlier life stages (eggs and larvae) are being intercepted at point of inspection most frequently, challenging the possibilities of morphological identification. To realize a rapid and reliable identification for all stages, we developed and validated four simplex real-time polymerase chain reaction identification tests based on the mitochondrial cytochrome b gene using dual-labeled hydrolysis probes. Method validation on dilutions of extracted DNA of the target organisms showed that low levels of template (up to 0.2-100 pg) can reliably be identified. No cross-reactivity was observed with 14 nontarget Spodoptera and 5 non-Spodoptera species in the specific Spodoptera tests. The tests showed to be repeatable, reproducible (both 100%), and robust. The new Spodoptera tests have proven to be suitable tools for routine identification of all life stages of S. eridania, S. frugiperda, S. littoralis, and S. litura. PMID:25195458

  8. Mortality Dynamics of Spodoptera frugiperda (Lepidoptera: Noctuidae) Immatures in Maize

    PubMed Central

    Varella, Andrea Corrêa; Menezes-Netto, Alexandre Carlos; Alonso, Juliana Duarte de Souza; Caixeta, Daniel Ferreira; Peterson, Robert K. D.; Fernandes, Odair Aparecido

    2015-01-01

    We characterized the dynamics of mortality factors affecting immature developmental stages of the fall armyworm, Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae). Multiple decrement life tables for egg and early larval stages of S. frugiperda in maize (Zea mays L.) fields were developed with and without augmentative releases of Telenomus remus Nixon (Hymenoptera: Platygastridae) from 2009 to 2011. Total egg mortality ranged from 73 to 81% and the greatest egg mortality was due to inviability, dislodgement, and predation. Parasitoids did not cause significant mortality in egg or early larval stages and the releases of T. remus did not increase egg mortality. Greater than 95% of early larvae died from predation, drowning, and dislodgment by rainfall. Total mortality due to these factors was largely irreplaceable. Results indicate that a greater effect in reducing generational survival may be achieved by adding mortality to the early larval stage of S. frugiperda. PMID:26098422

  9. Mortality Dynamics of Spodoptera frugiperda (Lepidoptera: Noctuidae) Immatures in Maize.

    PubMed

    Varella, Andrea Corrêa; Menezes-Netto, Alexandre Carlos; Alonso, Juliana Duarte de Souza; Caixeta, Daniel Ferreira; Peterson, Robert K D; Fernandes, Odair Aparecido

    2015-01-01

    We characterized the dynamics of mortality factors affecting immature developmental stages of the fall armyworm, Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae). Multiple decrement life tables for egg and early larval stages of S. frugiperda in maize (Zea mays L.) fields were developed with and without augmentative releases of Telenomus remus Nixon (Hymenoptera: Platygastridae) from 2009 to 2011. Total egg mortality ranged from 73 to 81% and the greatest egg mortality was due to inviability, dislodgement, and predation. Parasitoids did not cause significant mortality in egg or early larval stages and the releases of T. remus did not increase egg mortality. Greater than 95% of early larvae died from predation, drowning, and dislodgment by rainfall. Total mortality due to these factors was largely irreplaceable. Results indicate that a greater effect in reducing generational survival may be achieved by adding mortality to the early larval stage of S. frugiperda. PMID:26098422

  10. ANALYSIS OF THE 'SPODOPTERA FRUGIPERDA' NUCLEAR POLYHEDROSIS VIRUS GENOME BY RESTRICTION ENDONUCLEASES AND ELECTRON MICROSCOPY

    EPA Science Inventory

    Restriction endonuclease analysis was used to differentiate between four strains of Spodoptera frugiperda nuclear polyhedrosis virus from different geographical areas. In addition, partial denaturation was performed, and a partial denaturation map was constructed for the Ohio str...

  11. First microsatellites from Spodoptera frugiperda (Lepidoptera: Noctuidae) and their potential use for population genetics

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This is the first report of sequence-specific microsatellite markers (SSRs) of Spodoptera frugiperda (Smith) (Lepidoptera: Noctuidae), an economically important pest of the American continent. We developed 178 microsatellite markers using pyrosequencing, and screened 15 individuals from 8 isofamili...

  12. First microsatellites developed from Spodoptera frugiperda and their potential use for population genetics

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This is the first report of sequence-specific microsatellite markers (SSRs) of Spodoptera frugiperda (Smith) (Lepidoptera: Noctuidae), an economically important pest of the American continent. We developed 178 microsatellite markers using pyrosequencing, and screened 15 individuals from 8 colonies ...

  13. Population variation of the fall armyworm, Spodoptera frugiperda, in the Western Hemisphere

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae), the fall armyworm is the most economically important maize pest in the western hemisphere. This research focused on the genetic variability of the maize host strain because there is a lack of information in this area of S. frugiperda rese...

  14. Genomic sequence analysis of a fast-killing isolate of the Spodoptera frugiperda multiple nucleopolyhedrovirus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Six clones of Spodoptera frugiperda multiple nucleopolyhedrovirus (SfMNPV) were plaque-purified from field isolates collected in Missouri, USA. In bioassays, four of the plaque-purified isolates killed neonate S. frugiperda larvae more rapidly than the field isolates from which they were derived, w...

  15. 'SPODOPTERA FRUGIPERDA' NUCLEAR POLYHEDROSIS VIRUS GENOME: PHYSICAL MAPS FOR RESTRICTION ENDONUCLEASES BAMHI AND HINDIII

    EPA Science Inventory

    The physical map for the genome of Spodoptera frugiperda nuclear polyhedrosis virus was constructed for restriction endonucleases BamHI and HindIII. The ordering of the restriction fragments was accomplished by cross-blot hybridization of BamHI, HindIII, and EcoRI fragments. The ...

  16. Laboratory and field evaluations for efficacy of a fast-killing baculovirus isolate from Spodoptera frugiperda

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Three biopesticide parameters were evaluated for a fast-killing isolate (3AP2) Spodoptera frugiperda multiple nucleopolyhedrovirus (SfMNPV) and a wild-type isolate (Sf3) of the same baculovirus. Both isolates were evaluated for virus production using in vivo methods, for speed of kill based on bioas...

  17. Genetic variability of Spodoptera frugiperda (Lepidoptera: Noctuidae) in the Western Hemisphere

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The fall armyworm (FAW), Spodoptera frugiperda (J. E. Smith) is a migratory and polyphagous pest of both cultivated and uncultivated plant species in the Western Hemisphere. We investigated the genetic diversity of FAW by collecting 31 representative samples from the United States, Argentina, Panama...

  18. Rearing optimization of two races of the Fall armyworm Spodoptera frugiperda feeding natural host plants

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Two ecological races of the Fall armyworm Spodoptera frugiperda were raised under laboratory conditions feeding on natural host plants (corn and bermuda grass). Three rearing containers were used: a plastic container and a vertical cylinder to test fitness when feeding gregariously, and individual ...

  19. FLAVONOIDS OF ZOYSIAGRASS (ZOYSIA SPP) CULTIVARS VARYING IN FALL ARMYWORM (SPODOPTERA FRUGIPERDA)RESISTANCE

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Numerous cultivars of zoysiagrass (Zoysia spp) have been developed for the turf industry in the United States. These cultivars have varying degrees of biotic and abiotic stress tolerances. One pest of importance to zoysiagrass is the fall armyworm (Spodoptera frugiperda J.E. Smith). Cultivars suc...

  20. Genetic and biological variation among nucleopolyhedrovirus isolates from spodoptera frugiperda (lepidotpera: noctuidae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A PCR-based method was used to identify and distinguish among 40 uncharacterized nucleopolyhedrovirus (NPV) isolates from the moth Spodoptera frugiperda that were part of an insect virus collection. Phylogenetic analysis was carried out with sequences amplified from two strongly conserved loci (pol...

  1. Natural Distribution of Parasitoids of Larvae of the Fall Armyworm, Spodoptera frugiperda, in Argentina

    PubMed Central

    Gabriela Murúa, M.; Molina-Ochoa, Jaime; Fidalgo, Patricio

    2009-01-01

    To develop a better understanding of the natural distribution of the fall armyworm, Spodoptera frugiperda (Smith) (Lepidoptera: Noctuidae), and to update the knowledge of the incidence of its complex of parasitoids. S. frugiperda, samplings in whorl-stage corn were carried out in provinces of Argentina from 1999 to 2003. S. frugiperda larvae were collected from corn in localities of the provinces of Tucumán, Salta, Jujuy, Santiago del Estero, La Rioja, Córdoba, San Luis, Chaco and Misiones. In each locality 30 corn plants were sampled and only larvae located in those plants were collected. The parasitoids that emerged from S. frugiperda larvae were identified and counted. The abundance of the parasitoids and the parasitism rate were estimated. The S. frugiperda parasitoids collected were Campoletis grioti (Blanchard) (Hymenoptera: Ichneumonidae), Chelonus insularis (Cresson) (Hymenoptera: Braconidae), Archytas marmoratus (Townsend) (Diptera Tachinidae) and/or A. incertus (Macquart), Ophion sp. (Hymenoptera: Ichneumonidae), Euplectrus platyhypenae Howard (Hymenoptera: Eulophidae), and Incamyia chilensis (Aldrich) (Diptera Tachinidae). C. grioti was the most abundant and frequent during the five-year survey. Similar diversity of parasitoids was obtained in all the provinces, with the exception of I. chilensis and E. platyhypenae that were recovered only in the province of Salta. In the Northwestern region, in Tucumán, C. grioti and species of Archytas were the most abundant and frequent parasitoids. On the contrary, in Salta and Jujuy Ch. insularis was the parasitoid most abundant and frequently recovered. The parasitism rate obtained in Tucumán, Salta and Jujuy provinces were 21.96%, 17.87% and 6.63% respectively with an average of 18.93%. These results demonstrate that hymenopteran and dipteran parasitoids of S. frugiperda occurred differentially throughout the Argentinian provinces and played an important role on the natural control of the S. frugiperda larval

  2. Population variation of the fall armyworm, Spodoptera frugiperda, in the Western Hemisphere.

    PubMed

    Clark, Pete L; Molina-Ochoa, Jaime; Martinelli, Samuel; Skoda, Steven R; Isenhour, David J; Lee, Donald J; Krumm, Jeffrey T; Foster, John E

    2007-01-01

    Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae), the fall armyworm is the most economically important maize pest in the western hemisphere. This research focused on the genetic variability of the maize host strain because there is a lack of information in this area of S. frugiperda research. Amplified fragment length polymorphism (AFLP) was used to assess the genetic variability of S. frugiperda over a large geographic area. Twenty populations were collected from the maize, one population was collected from princess tree, one population was collected from lemon tree, and one population was collected from bermudagrass. The 23 populations were from Mexico, the continental United States, Puerto Rico, Brazil, and Argentina. The objective of this research was to evaluate whether the majority of genetic variability was within populations or between populations. The AFLP results showed that the majority of the genetic variability is within populations and not between populations, indicating minor gene flow and suggesting that S. frugiperda in the Western Hemisphere are an interbreeding population. PMID:20334595

  3. Population Variation of the Fall Armyworm, Spodoptera frugiperda, in the Western Hemisphere

    PubMed Central

    Clark, Pete L.; Molina-Ochoa, Jaime; Martinelli, Samuel; Skoda, Steven R.; Isenhour, David J.; Lee, Donald J.; Krumm, Jeffrey T.; Foster, John E.

    2007-01-01

    Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae), the fall armyworm is the most economically important maize pest in the western hemisphere. This research focused on the genetic variability of the maize host strain because there is a lack of information in this area of S. frugiperda research. Amplified fragment length polymorphism (AFLP) was used to assess the genetic variability of S. frugiperda over a large geographic area. Twenty populations were collected from the maize, one population was collected from princess tree, one population was collected from lemon tree, and one population was collected from bermudagrass. The 23 populations were from Mexico, the continental United States, Puerto Rico, Brazil, and Argentina. The objective of this research was to evaluate whether the majority of genetic variability was within populations or between populations. The AFLP results showed that the majority of the genetic variability is within populations and not between populations, indicating minor gene flow and suggesting that S. frugiperda in the Western Hemisphere are an interbreeding population. PMID:20334595

  4. Phosphorylation of the insect immunophilin FKBP46 by the Spodoptera frugiperda homolog of casein kinase II.

    PubMed

    Steplewski, A; Ebel, W; Planey, S L; Alnemri, E S; Robertson, N M; Litwack, G

    2000-04-01

    Immunophilins are a family of conserved proteins found in both prokaryotes and eukaryotes, that exhibit peptidylprolyl isomerase (PPIase) activity. Members of this family bind to immunosuppressive drugs and on this basis are divided into two classes: FKBPs bind to FK506 and rapamycin, while cyclophilins bind to cyclosporin A. In this paper, we report on insect immunophilin FKBP46 and its associated kinase. The insect FKBP46 belongs to the high-molecular-weight immunophilins and shares many characteristic features with its mammalian counterparts, but its functional role remains unclear. Here, we show that FKBP46 is phosphorylated by a protein kinase present in the nucleus of both insect Spodoptera frugiperda (Sf9) and human Jurkat cells. This protein kinase is immunoreactive with polyclonal antiserum raised against Drosophila melanogaster casein kinase II (CKII). We have cloned, overexpressed and characterized a new member of the CKII family derived from Spodoptera frugiperda cells. Recombinant Sf9 CKII alpha subunit shares 75% identity to human, chicken and Drosophila melanogaster homologs, whereas the Sf9 CKII beta subunit is 77% identical to rat, chicken and human. Moreover, we demonstrate that the insect immunophilin FKBP46 can be phosphorylated by human and Sf9 casein kinase II. Finally, we show that FKBP46 interacts with DNA, and this interaction is not prevented by phosphorylation. PMID:10767538

  5. Differential Protection of Cry1Fa Toxin against Spodoptera frugiperda Larval Gut Proteases by Cadherin Orthologs Correlates with Increased Synergism

    PubMed Central

    Rahman, Khalidur; Abdullah, Mohd Amir F.; Ambati, Suresh; Taylor, Milton D.

    2012-01-01

    The Cry proteins produced by Bacillus thuringiensis (Bt) are the most widely used biopesticides effective against a range of crop pests and disease vectors. Like chemical pesticides, development of resistance is the primary threat to the long-term efficacy of Bt toxins. Recently discovered cadherin-based Bt Cry synergists showed the potential to augment resistance management by improving efficacy of Cry toxins. However, the mode of action of Bt Cry synergists is thus far unclear. Here we elucidate the mechanism of cadherin-based Cry toxin synergism utilizing two cadherin peptides, Spodoptera frugiperda Cad (SfCad) and Manduca sexta Cad (MsCad), which differentially enhance Cry1Fa toxicity to Spodoptera frugiperda neonates. We show that differential SfCad- and MsCad-mediated protection of Cry1Fa toxin in the Spodoptera frugiperda midgut correlates with differential Cry1Fa toxicity enhancement. Both peptides exhibited high affinity for Cry1Fa toxin and an increased rate of Cry1Fa-induced pore formation in S. frugiperda. However, only SfCad bound the S. frugiperda brush border membrane vesicle and more effectively prolonged the stability of Cry1Fa toxin in the gut, explaining higher Cry1Fa enhancement by this peptide. This study shows that cadherin fragments may enhance B. thuringiensis toxicity by at least two different mechanisms or a combination thereof: (i) protection of Cry toxin from protease degradation in the insect midgut and (ii) enhancement of pore-forming ability of Cry toxin. PMID:22081566

  6. Geographic variation in sexual attraction of Spodoptera frugiperda corn- and rice-strain males to pheromone lures

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The corn- and rice-strains of Spodoptera frugiperda exhibit several genetic and behavioral differences and appear to be undergoing ecological speciation in sympatry. Previous studies reported conflicting results when investigating male attraction to pheromone lures in different regions, but this cou...

  7. Spatial genetic variation among Spodoptera frugiperda (Lepidoptera: Noctuidae) sampled from the United States, Puerto Rico, Panama, and Argentina

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The fall armyworm (FAW), Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae), is a migratory and polyphagous pest of both cultivated and uncultivated plant species in the Western Hemisphere. Understanding the genetic diversity and gene flow of this economically important pest can help to de...

  8. Occurrence of entomopathogenic fungi and parasitic nematodes on Spodoptera frugiperda (Lepidoptera: Noctuidae) larvae collected in Central Chiapas, Mexico

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fall armyworm larvae (FAW), Spodoptera frugiperda (J. E. Smith) were collected from whorl-stage cornfields, between the V2 and V4 stages, in 22 localities of Central, Chiapas, México, called "La Frailesca" during late June 2009 to determine the occurrence of native entomopathogens and parasitic nema...

  9. Lyophilization of Spodoptera frugiperda (Lepidoptera: Noctuidae) larvae yields high-quality DNA for use in AFLP genetic studies

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Agricultural research in the 21st century has become a collaborative effort. Research on crop pests like Spodoptera frugiperda (J.E. Smith), commonly known as the fall armyworm (FAW), can involve international collaboration because it is a pest not only in the southern United States, but also in La...

  10. COMPARISON OF BIOASSAY AND ENZYME-LINKED IMMUNOSORBENT ASSAY FOR QUANTIFICATION OF 'SPODOPTERA FRUGIPERDA' NUCLEAR POLYHEDROSIS VIRUS IN SOIL

    EPA Science Inventory

    Standard curves with known amounts of Spodoptera frugiperda nuclear polyhedrosis virus (NPV) in soil were established with a bioassay and with an enzyme-linked immunosorbent assay (ELISA). The bioassay detected as few as 4 x 10 to the 4th power polyhedral inclusion bodies (PIB)/g...

  11. Toxic effects of Citrus aurantium and C. limon essential oils on Spodoptera frugiperda (Lepidoptera: Noctuidae).

    PubMed

    Villafañe, Emilio; Tolosa, Diego; Bardón, Alicia; Neske, Adriana

    2011-09-01

    Citrus aurantium and C. limon were selected in the search for natural plant insecticides. The essential oils of C. aurantium and C. limon and ethanol extracts of the seeds, pulp, albedo, and peel of C. aurantium were incorporated into the larval diet of the lepidopteran pest Spodoptera frugiperda. Larval and pupal mortality were quantified and adult malformation was observed. C aurantium essential oil had antifeedant action and the mixture of albedo ethanol extract and C aurantium essential oil had toxic effects on S. frugiperda larvae at early stages, when they had not yet produced major damage to the crop. Our results indicated that a mixture of ethanol extract of albedo and C. aurantium essential oil (250 microg of extract mix per g of diet) deterred feeding by 46% and had the highest larval mortality (100%) of the materials tested. The peel extract (250 microg per g of diet) produced an increment in growth rate and diet consumption. However, 40% of the larval and 45% of the pupal populations died after 96 h of treatment. The blend of essential oil and C. aurantium albedo ethanol extract showed the lowest consumption and a poor nutrient conversion into biomass. Finally, the presence of D-limonene and nootkatone in the peel ethanol extract, and C. limon and C. aurantium essential oils, may be the cause of the response in the feeding behavior and toxic effects found on S. frugiperda. PMID:21941921

  12. Inhibition of the responses to sex pheromone of the fall armyworm, Spodoptera frugiperda.

    PubMed

    Malo, Edi A; Rojas, Julio C; Gago, Rafael; Guerrero, Ángel

    2013-01-01

    Trifluoromethyl ketones reversibly inhibit pheromone-degrading esterases in insect olfactory tissues, affecting pheromone detection and behavior of moth males. In this work, (Z)-9-tetradecenyl trifluoromethyl ketone (Z9-14:TFMK), a closely-related analogue of the pheromone of the fall armyworm, Spodoptera frugiperda (Smith) (Lepidoptera: Noctuidae), was prepared and tested in electroantennogram and field tests as possible inhibitors of the pheromone action. The electroantennogram parameters, amplitude, and the repolarization time of the antennal responses of S. frugiperda males were affected by Z9-14:TFMK vapors. Exposure of male antennae to a stream of air passing through 100 μg of the ketone produced a significant reduction of the amplitude and an increase of 2/3 repolarization time signals to the pheromone. The effect was reversible and dose-dependent. In the field, the analogue significantly decreased the number of males caught when mixed with the pheromone in 10:1 ratio. The results suggest that Z9-14:TFMK is a mating disruptant of S. frugiperda and may be a good candidate to consider in future strategies to control this pest. PMID:24766416

  13. Cytochrome P450s from the fall armyworm (Spodoptera frugiperda): responses to plant allelochemicals and pesticides.

    PubMed

    Giraudo, M; Hilliou, F; Fricaux, T; Audant, P; Feyereisen, R; Le Goff, G

    2015-02-01

    Spodoptera frugiperda is a polyphagous lepidopteran pest that encounters a wide range of toxic plant metabolites in its diet. The ability of this insect to adapt to its chemical environment might be explained by the action of major detoxification enzymes such as cytochrome P450s (or CYP). Forty-two sequences coding for P450s were identified and most of the transcripts were found to be expressed in the midgut, Malpighian tubules and fat body of S. frugiperda larvae. Relatively few P450s were expressed in the established cell line Sf9. In order to gain information on how these genes respond to different chemical compounds, larvae and Sf9 cells were exposed to plant secondary metabolites (indole, indole-3-carbinol, quercetin, 2-tridecanone and xanthotoxin), insecticides (deltamethrin, fipronil, methoprene, methoxyfenozide) or model inducers (clofibrate and phenobarbital). Several genes were induced by plant chemicals such as P450s from the 6B, 321A and 9A subfamilies. Only a few genes responded to insecticides, belonging principally to the CYP9A family. There was little overlap between the response in vivo measured in the midgut and the response in vitro in Sf9 cells. In addition, regulatory elements were detected in the promoter region of these genes. In conclusion, several P450s were identified that could potentially be involved in the adaptation of S. frugiperda to its chemical environment. PMID:25315858

  14. Inhibition of the Responses to Sex Pheromone of the Fall Armyworm, Spodoptera frugiperda

    PubMed Central

    Malo, Edi A.; Rojas, Julio C.; Gago, Rafael; Guerrero, Ángel

    2013-01-01

    Trifluoromethyl ketones reversibly inhibit pheromone-degrading esterases in insect olfactory tissues, affecting pheromone detection and behavior of moth males. In this work, (Z)-9-tetradecenyl trifluoromethyl ketone (Z9-14:TFMK), a closely-related analogue of the pheromone of the fall armyworm, Spodoptera frugiperda (Smith) (Lepidoptera: Noctuidae), was prepared and tested in electroantennogram and field tests as possible inhibitors of the pheromone action. The electroantennogram parameters, amplitude, and the repolarization time of the antennal responses of S. frugiperda males were affected by Z9-14:TFMK vapors. Exposure of male antennae to a stream of air passing through 100 ìg of the ketone produced a significant reduction of the amplitude and an increase of 2/3 repolarization time signals to the pheromone. The effect was reversible and dose-dependent. In the field, the analogue significantly decreased the number of males caught when mixed with the pheromone in 10:1 ratio. The results suggest that Z9-14:TFMK is a mating disruptant of S. frugiperda and may be a good candidate to consider in future strategies to control this pest. PMID:24766416

  15. Identification and characteristics of microRNAs from army worm, Spodoptera frugiperda cell line Sf21.

    PubMed

    Kakumani, Pavan Kumar; Chinnappan, Mahendran; Singh, Ashok K; Malhotra, Pawan; Mukherjee, Sunil K; Bhatnagar, Raj K

    2015-01-01

    microRNAs play important regulatory role in all intrinsic cellular functions. Amongst lepidopteran insects, miRNAs from only Bombyx mori have been studied extensively with a little focus on Spodoptera sp. In the present study, we identified a total of 226 miRNAs from Spodoptera frugiperda cell line Sf21. Of the total, 116 miRNAs were well conserved within other insects, like B. mori, Drosophila melanogaster and Tribolium castenum while the remaining 110 miRNAs were identified as novel based on comparative analysis with the insect miRNA data set. Landscape distribution analysis based on Sf21 genome assembly revealed clustering of few novel miRNAs. A total of 5 miRNA clusters were identified and the largest one encodes 5 miRNA genes. In addition, 12 miRNAs were validated using northern blot analysis and putative functional role assignment for 6 Sf miRNAs was investigated by examining their relative abundance at different developmental stages of Spodoptera litura and body parts of 6th instar larvae. Further, we identified a total of 809 potential target genes with GO terms for selected miRNAs, involved in different metabolic and signalling pathways of the insect. The newly identified miRNAs greatly enrich the repertoire of insect miRNAs and analysis of expression profiles reveal their involvement at various steps of biochemical pathways of the army worm. PMID:25693181

  16. F2 screen for resistance to Bacillus thuringiensis Cry2Ab2-maize in field populations of Spodoptera frugiperda (Lepidoptera: Noctuidae) from the southern United States

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The fall armyworm, Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae), is a target of transgenic maize and cotton expressing Bacillus thuringiensis (Bt) proteins in both North and South America. In 2013 and 2014, a total of 215 F2 two-parent families of S. frugiperda were established usin...

  17. Investigating the Molecular Mechanisms of Organophosphate and Pyrethroid Resistance in the Fall Armyworm Spodoptera frugiperda

    PubMed Central

    Carvalho, Renato A.; Omoto, Celso; Field, Linda M.; Williamson, Martin S.; Bass, Chris

    2013-01-01

    The fall armyworm Spodoptera frugiperda is an economically important pest of small grain crops that occurs in all maize growing regions of the Americas. The intensive use of chemical pesticides for its control has led to the selection of resistant populations, however, to date, the molecular mechanisms underlying resistance have not been characterised. In this study the mechanisms involved in the resistance of two S. frugiperda strains collected in Brazil to chlorpyrifos (OP strain) or lambda-cyhalothrin (PYR strain) were investigated using molecular and genomic approaches. To examine the possible role of target-site insensitivity the genes encoding the organophosphate (acetylcholinesterase, AChE) and pyrethroid (voltage-gated sodium channel, VGSC) target-site proteins were PCR amplified. Sequencing of the S. frugiperda ace-1 gene identified several nucleotide changes in the OP strain when compared to a susceptible reference strain (SUS). These result in three amino acid substitutions, A201S, G227A and F290V, that have all been shown previously to confer organophosphate resistance in several other insect species. Sequencing of the gene encoding the VGSC in the PYR strain, identified mutations that result in three amino acid substitutions, T929I, L932F and L1014F, all of which have been shown previously to confer knockdown/super knockdown-type resistance in several arthropod species. To investigate the possible role of metabolic detoxification in the resistant phenotype of the OP and PYR stains all EST sequences available for S. frugiperda were used to design a gene-expression microarray. This was then used to compare gene expression in the resistant strains with the susceptible reference strain. Members of several gene families, previously implicated in metabolic resistance in other insects were found to be overexpressed in the resistant strains including glutathione S-transferases, cytochrome P450s and carboxylesterases. Taken together these results provide

  18. Growth inhibitory effects on fall armyworm Spodoptera frugiperda of some limonoids isolated from Cedrela spp. (Meliaceae).

    PubMed

    Céspedes, C L; Calderón, J S; Lina, L; Aranda, E

    2000-05-01

    Dichloromethane extracts of Cedrela salvadorensis and Cedrela dugessi afforded a photogedunin epimeric mixture, gedunin and cedrelanolide. These compounds and the photogedunin epimeric acetates 3 and 4 at the 23-OH position were evaluated against Spodoptera frugiperda. Toosendanin, isolated from Melia azedarach, was used as a positive control. When tested for activity on neonate larvae into the no-choice bioassays, gedunin, photogedunin epimeric mixture, and photogedunin acetates mixture caused significant larval mortality with LC(50) of 39.0, 10.0, and 8.0 ppm at 7 days, respectively, as well as growth reduction. All the compounds tested inhibited larval growth, compared to the control, in a concentration-dependent manner. In addition, it was possible to observe significant reduced pupal weights and adult emergence. All the tested compounds except cedrelanolide showed comparable activity to that of toosendanin. PMID:10820113

  19. Vip3Aa induces apoptosis in cultured Spodoptera frugiperda (Sf9) cells.

    PubMed

    Jiang, Kun; Mei, Si-Qi; Wang, Ting-Ting; Pan, Jin-Hua; Chen, Yue-Hua; Cai, Jun

    2016-09-15

    The vegetative insecticidal proteins (Vip) secreted by many Bacillus thuringiensis strains during their vegetative growth stage are regarded as second generation insecticidal proteins, as they share no sequence or structural homology with known crystal insecticidal proteins (Cry) and have a broad insecticidal spectrum. Compared with insecticidal crystal proteins (ICPs), the insecticidal mechanisms of Vips have been little studied. Here we investigated the mechanism responsible for Vip3Aa toxicity in cultured insect cells. Using, flow cytometry analyzes, TUNEL staining and DNA fragmentation assays, we show that Vip3Aa can induce apoptosis in Spodoptera frugiperda (Sf9) cells and cause cells to arrest at the G2/M phase. We also show that Vip3Aa can disrupt mitochondrial membrane potential (ΔΨm), leading to the activation of Sf-caspase-1, suggesting that a mitochondrial mediated and caspase dependent pathway may be involved in Vip3Aa-induced apoptosis in Sf9 cells. PMID:27476462

  20. Toxicity of selected tremorgenic mycotoxins and related compounds to Spodoptera frugiperda and Heliothis zea.

    PubMed

    Dowd, P F; Cole, R J; Vesonder, R F

    1988-12-01

    A series of tremorgenic mycotoxins and related compounds were tested for oral toxicity to the fall armyworm (Spodoptera frugiperda) and corn earworm (Heliothis zea) by incorporation of materials into artificial diets and examining mortality and weights after 7 days. Significant mortality to both insect species was caused with dihydroxyaflavinine and roseotoxin B, while significant mortality to H. zea was also caused by penitrem A at 25 ppm. After 7 days, weighs of larvae treated with 25 ppm penitrem A, roseotoxin B, and verruculogen were less than 50% of controls for both insect species. Weights of H. zea larvae treated with 25 ppb of penitrem A were less than 50% those of control larvae. Relative toxicities of the tremorgens and related compounds to insects compared to vertebrates are discussed. PMID:3209479

  1. The Complete Sequence of the First Spodoptera frugiperda Betabaculovirus Genome: A Natural Multiple Recombinant Virus

    PubMed Central

    Cuartas, Paola E.; Barrera, Gloria P.; Belaich, Mariano N.; Barreto, Emiliano; Ghiringhelli, Pablo D.; Villamizar, Laura F.

    2015-01-01

    Spodoptera frugiperda (Lepidoptera: Noctuidae) is a major pest in maize crops in Colombia, and affects several regions in America. A granulovirus isolated from S. frugiperda (SfGV VG008) has potential as an enhancer of insecticidal activity of previously described nucleopolyhedrovirus from the same insect species (SfMNPV). The SfGV VG008 genome was sequenced and analyzed showing circular double stranded DNA of 140,913 bp encoding 146 putative ORFs that include 37 Baculoviridae core genes, 88 shared with betabaculoviruses, two shared only with betabaculoviruses from Noctuide insects, two shared with alphabaculoviruses, three copies of own genes (paralogs) and the other 14 corresponding to unique genes without representation in the other baculovirus species. Particularly, the genome encodes for important virulence factors such as 4 chitinases and 2 enhancins. The sequence analysis revealed the existence of eight homologous regions (hrs) and also suggests processes of gene acquisition by horizontal transfer including the SfGV VG008 ORFs 046/047 (paralogs), 059, 089 and 099. The bioinformatics evidence indicates that the genome donors of mentioned genes could be alpha- and/or betabaculovirus species. The previous reported ability of SfGV VG008 to naturally co-infect the same host with other virus show a possible mechanism to capture genes and thus improve its fitness. PMID:25609309

  2. Identification and Characterization of Pathogen-Response Genes (repat) in Spodoptera frugiperda (Lepidoptera: Noctuidae).

    PubMed

    Machado, Vilmar; Serrano, Jose; Galián, Jose

    2016-01-01

    The fall armyworm (Spodoptera frugiperda, Noctuidae, Lepidoptera) is one of the most important crop pests in the Americas, causing significant damage to maize, rice and sorghum. The mechanisms that determine its defences against pathogens are particularly relevant for the development of management and control strategies. We used an in silico approach to identify and characterize pathogen response genes (repat) present in different tissue libraries of S. fugiperda. The analyses revealed complete cDNA for nine repat genes; of these, repat15 and repat39 were found in libraries from a specific tissue--the midgut of larvae fed with xenobiotic substances. High expression levels of some genes were found in different libraries: 39 hits in repat30 in challenged hemocytes, 16 hits in repat31 in fat body, 10 hits in repat32 in fat body and 10 in challenged hemocytes, and 10 hits in repat38 in midgut of non-treated larvae and midgut of larvae fed with natural and xenobiotic substances. The genes corresponded to two ontology categories, stress response and immune response, and their phylogenetic relationships, nucleotide similarity, number of amino acid residues and molecular weights agree with what has been described for repat genes. It is noteworthy that proteins encoded by the repat genes of S. frugiperda have important defence functions in other tissues beyond midgut and that their functional categories are likely diverse, as they are related to cell envelope structure, energy metabolism, transport and binding. PMID:27172709

  3. Spodoptera frugiperda (J.E. Smith) with field-evolved resistance to Bt maize are susceptible to Bt pesticides.

    PubMed

    Jakka, S R K; Knight, V R; Jurat-Fuentes, J L

    2014-10-01

    Field-evolved resistance to maize event TC1507 expressing the Cry1Fa toxin from Bacillus thuringiensis (Bt) was detected in populations of Spodoptera frugiperda from Puerto Rico. We tested for cross-resistance to purified Cry1A toxins and commercial Bt pesticides in susceptible (Benzon) and TC1507-resistant (456) strains of S. frugiperda. Larvae from the 456 strain exhibited cross-resistance to Cry1Ab and Cry1Ac toxins, while no differences in susceptibility to XenTari WG and DiPel ES pesticides were detected. These data support cross-resistance to toxins that share binding sites with Cry1Fa and no cross-resistance to Bt pesticides in S. frugiperda with field-evolved resistance to Bt maize. PMID:25218399

  4. Susceptibility and aversion of Spodoptera frugiperda (Lepidoptera: Noctuidae) to Cry1F Bt maize and considerations for insect resistance management.

    PubMed

    Binning, Rachel R; Coats, Joel; Kong, Xiaoxiao; Hellmich, Richard L

    2014-02-01

    Bacillus thuringiensis (Bt) maize was developed primarily for North American pests such as European corn borer (Ostrinia nubilalis (Hübner)). However, most Bt maize products are also cultivated outside of North America, where the primary pests may be different and may have lower susceptibility to Bt toxins. Fall armyworm (Spodoptera frugiperda JE Smith) is an important pest and primary target of Bt maize in Central and South America. S. frugiperda susceptibility to Cry1F (expressed in event TC1507) is an example of a pest-by-toxin interaction that does not meet the high-dose definition. In this study, the behavioral and toxic response of S. frugiperda to Cry1F maize was investigated by measuring the percentage of time naive third instars spent feeding during a 3-min exposure. S. frugiperda also were exposed as third instars to Cry1F maize for 14 d to measure weight gain and survival. S. frugiperda demonstrated an initial, postingestive aversive response to Cry1F maize, and few larvae survived the 14 d exposure. The role of susceptibility and avoidance are discussed in the context of global IRM refuge strategy development for Bt products. PMID:24665722

  5. Attraction of the parasitoid Cotesia marginiventris to host (Spodoptera frugiperda) frass is affected by transgenic maize.

    PubMed

    Desneux, Nicolas; Ramírez-Romero, Ricardo; Bokonon-Ganta, Aimé H; Bernal, Julio S

    2010-10-01

    We assessed in the laboratory the attraction of the parasitoid Cotesia marginiventris (Cresson) toward odors emitted by conventional maize (Zea mays L. ssp. mays) and Bt (Bacillus thuringiensis) maize seedlings following actual or simulated injury by Spodoptera frugiperda (Smith), the parasitoid's host, and emitted by the host's frass, produced following consumption of conventional or Bt maize seedlings. Females of C. marginiventris exhibited similarly strong responses to conventional and Bt maize seedlings injured by the host or with simulated injury, and these were stronger than responses to clean air. In contrast, the responses of C. marginiventris females were consistently weaker toward host frass derived from Bt maize tissue compared to frass derived from conventional maize tissue. We hypothesized that the weakened response was due to a detrimental effect of Bt endotoxins, present in the Bt maize tissue, on the bacterial community present in the host's gut and frass, including bacteria that produce odors attractive to C. marginiventris. As an initial test of our hypothesis, we compared between the responses of C. marginiventris females to host frass produced following consumption of Bt maize and frass produced from conventional maize which had been treated with an antibiotic (tetracycline) to eliminate host gut bacteria. Our results showed that C. marginiventris females responded similarly weakly to host frass derived from conventional maize tissue treated with antibiotic and to frass derived from Bt maize tissue, treated or untreated with antibiotic, while they responded strongly to frass derived from conventional maize untreated with antibiotic, so provided initial, partial support for our hypothesis. We discussed the weakened response of C. marginiventris females to host frass derived from Bt maize in the context of plausible impacts of transgenic crop cultivars on parasitoid foraging and populations, and the implications for biological control of non

  6. A COMPREHENSIVE STUDY ON APOPTOSIS INDUCTION BY AZADIRACHTIN IN Spodoptera frugiperda CULTURED CELL LINE Sf9.

    PubMed

    Shu, Benshui; Wang, Wenxiang; Hu, Qingbo; Huang, Jingfei; Hu, Meiying; Zhong, Guohua

    2015-07-01

    The induction of apoptosis by azadirachtin, a well-known botanical tetranortriterpenoid isolated from the neem tree (Azadirachta indica A. Juss) and other members of the Meliaceae, was investigated in Spodoptera frugiperda cultured cell line (Sf9). Morphological changes in Sf9 cells treated by various concentrations of azadirachtin were observed at different times under light microscopy. Morphological and biochemical analysis indicated that Sf9 cells treated by 1.5 μg/mL azadirachtin showed typical morphological changes, which were indicative of apoptosis and a clear DNA ladder. The flow cytometry analysis showed the apoptosis rate reached a maximum value of 32.66% at 24 h with 1.5 μg/mL azadirachtin in Sf9 cells. The inhibition of Sf9 cell proliferation suggested that the effect of azadirachtin was dose dependent and the EC50 at 48 and 72 h was 2.727 × 10(-6) and 6.348 × 10(-9) μg/mL, respectively. The treatment of azadirachtin in Sf9 cells could significantly increase the activity of Sf caspase-1, but showed no effect on the activity of Topo I, suggesting that the apoptosis induced by azadirachtinin Sf9 cells is through caspase-dependent pathway. These results provided not only a series of morphological, biochemical, and toxicological comprehensive evidences for induction of apoptosis by azadirachtin, but also a reference model for screening insect cell apoptosis inducers from natural compounds. PMID:25828604

  7. Sublethal Effects of Essential Oils From Eucalyptus staigeriana (Myrtales: Myrtaceae), Ocimum gratissimum (Lamiales: Laminaceae), and Foeniculum vulgare (Apiales: Apiaceae) on the Biology of Spodoptera frugiperda (Lepidoptera: Noctuidae).

    PubMed

    Cruz, G S; Wanderley-Teixeira, V; Oliveira, J V; Lopes, F S C; Barbosa, D R S; Breda, M O; Dutra, K A; Guedes, C A; Navarro, D M A F; Teixeira, A A C

    2016-04-01

    Spodoptera frugiperda (Smith 1797) (Lepidoptera: Noctuidae) is a major pest of maize, Zea mays L. Its control is often achieved through repeated applications per season of insecticides, which may lead to adverse effects on the ecosystem. Thus, the study of alternative methods with less environmental impact has expanded to include the use of essential oils. These oils are products of the secondary metabolism in plants, and their insecticidal activity has been widely demonstrated in populations of many pest insects. This study evaluated the insecticidal activities of essential oils from Eucalyptus staigeriana, Ocimum gratissimum, and Foeniculum vulgare on Spodoptera frugiperda. Gas chromatography–mass spectrometry profiles and contact toxicity of these oils as well as their sublethal effects on larvae and reproductive parameters in adults were evaluated. All three oils had sublethal effects on S. frugiperda; however, the oil of O. gratissimum showed the best results at all doses tested. These essential oils may have promise for control of S. frugiperda. PMID:26868417

  8. Frequency of Cry1F Non-Recessive Resistance Alleles in North Carolina Field Populations of Spodoptera frugiperda (Lepidoptera: Noctuidae).

    PubMed

    Li, Guoping; Reisig, Dominic; Miao, Jin; Gould, Fred; Huang, Fangneng; Feng, Hongqiang

    2016-01-01

    Fall armyworm, Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae), is a target species of transgenic corn (Zea mays L.) that expresses single and pyramided Bacillus thuringiensis (Bt) toxin. In 2014, S. frugiperda were collected from a light trap in North Carolina, and a total of 212 F1/F2 isofemale lines of S. frugiperda were screened for resistance to Bt and non-Bt corn. All of the 212 isolines were susceptible to corn tissue expressing Cry1A.105 + Cry2Ab, Cry1F + Cry1A.105 + Cry2Ab, and Cry1F + Cry1Ab + Vip3Aa20. Growth rate bioassays were performed to isolate non-recessive Bt resistance alleles. Seven individuals out of the 212 isofemale lines carried major non-recessive alleles conferring resistance to Cry1F. A pooled colony was created from the seven individuals. This colony was 151.21 times more resistant to Cry1F than a known-susceptible population and was also resistant to Cry1A.105, but was not resistant to Cry2Ab and Vip3Aa20. The results demonstrate that field populations of S. frugiperda collected from North Carolina are generally susceptible to Cry1F, but that some individuals carry resistant alleles. The data generated in this study can be used as baseline data for resistance monitoring. PMID:27119741

  9. Frequency of Cry1F Non-Recessive Resistance Alleles in North Carolina Field Populations of Spodoptera frugiperda (Lepidoptera: Noctuidae)

    PubMed Central

    Li, Guoping; Reisig, Dominic; Miao, Jin; Gould, Fred; Huang, Fangneng; Feng, Hongqiang

    2016-01-01

    Fall armyworm, Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae), is a target species of transgenic corn (Zea mays L.) that expresses single and pyramided Bacillus thuringiensis (Bt) toxin. In 2014, S. frugiperda were collected from a light trap in North Carolina, and a total of 212 F1/F2 isofemale lines of S. frugiperda were screened for resistance to Bt and non-Bt corn. All of the 212 isolines were susceptible to corn tissue expressing Cry1A.105 + Cry2Ab, Cry1F + Cry1A.105 + Cry2Ab, and Cry1F + Cry1Ab + Vip3Aa20. Growth rate bioassays were performed to isolate non-recessive Bt resistance alleles. Seven individuals out of the 212 isofemale lines carried major non-recessive alleles conferring resistance to Cry1F. A pooled colony was created from the seven individuals. This colony was 151.21 times more resistant to Cry1F than a known-susceptible population and was also resistant to Cry1A.105, but was not resistant to Cry2Ab and Vip3Aa20. The results demonstrate that field populations of S. frugiperda collected from North Carolina are generally susceptible to Cry1F, but that some individuals carry resistant alleles. The data generated in this study can be used as baseline data for resistance monitoring. PMID:27119741

  10. Citrus limonoids and their semisynthetic derivatives as antifeedant agents against Spodoptera frugiperda larvae. A structure-activity relationship study.

    PubMed

    Ruberto, Giuseppe; Renda, Agatino; Tringali, Corrado; Napoli, Edoardo M; Simmonds, Monique S J

    2002-11-01

    The antifeedant activity of Citrus-derived limonoids limonin (1), nomilin (2), and obacunone (3) and their semisynthetic derivatives 4-26 was evaluated against a commercially important pest, Spodoptera frugiperda. Simple chemical conversions were carried out on the natural limonoids obtained from seeds of Citrus limon. These conversions focused on functional groups considered to be important for the biological activity, namely the C-7 carbonyl and the furan ring. In particular, reduction at C-7 afforded the related alcohols, and from these their acetates, oximes, and methoximes were prepared. Hydrogenation of the furan ring was also performed on limonin and obacunone. The known antifeedant properties of the Citrus limonoids are confirmed. Comparison with previously reported data shows that insect species vary in their behavioral responses to these structural modifications. Highly significant antifeedant activity (P < 0.01) for two natural (1 and 3) and three semisynthetic limonoids (4, 8, and 10) was observed against S. frugiperda. PMID:12405773

  11. Life-History Traits of Spodoptera frugiperda Populations Exposed to Low-Dose Bt Maize

    PubMed Central

    Sousa, Fernanda F.; Mendes, Simone M.; Santos-Amaya, Oscar F.; Araújo, Octávio G.; Oliveira, Eugenio E.

    2016-01-01

    Exposure to Bacillus thuringiensis (Bt) toxins in low- and moderate-dose transgenic crops may induce sublethal effects and increase the rate of Bt resistance evolution, potentially compromising control efficacy against target pests. We tested this hypothesis using the fall armyworm Spodoptera frugiperda, a major polyphagous lepidopteran pest relatively tolerant to Bt notorious for evolving field-relevant resistance to single-gene Bt maize. Late-instar larvae were collected from Bt Cry1Ab and non-Bt maize fields in five locations in Brazil, and their offspring was compared for survival, development, and population growth in rearing environment without and with Cry1Ab throughout larval development. Larval survival on Cry1Ab maize leaves varied from 20 to 80% among the populations. Larvae reared on Cry1Ab maize had seven-day delay in development time in relation to control larvae, and such delay was shorter in offspring of armyworms from Cry1Ab maize. Population growth rates were 50–70% lower for insects continuously exposed to Cry1Ab maize relative to controls, showing the population-level effect of Cry1Ab, which varied among the populations and prior exposure to Cry1Ab maize in the field. In three out of five populations, armyworms derived from Bt maize reared on Cry1Ab maize showed higher larval weight, faster larval development and better reproductive performance than the armyworms derived from non-Bt maize, and one of these populations showed better performance on both Cry1Ab and control diets, indicating no fitness cost of the resistance trait. Altogether, these results indicate that offspring of armyworms that developed on field-grown, single-gene Bt Cry1Ab maize had reduced performance on Cry1Ab maize foliage in two populations studied, but in other three populations, these offspring had better overall performance on the Bt maize foliage than that of the armyworms from non-Bt maize fields, possibly because of Cry1Ab resistance alleles in these populations

  12. Characterization of native Bacillus thuringiensis strains and selection of an isolate active against Spodoptera frugiperda and Peridroma saucia.

    PubMed

    Alvarez, Analía; Virla, Eduardo G; Pera, Licia M; Baigorí, Mario D

    2009-12-01

    Twelve Bacillus thuringiensis (Bt) strains, isolated from larvae and soil samples in Argentina, were molecularly and phenotypically characterized and their insecticidal activities against Spodoptera frugiperda and Peridroma saucia were determined. One isolate--Bt RT--produced more than 93% mortality on first instar larvae of both species, which was higher than that produced by the reference strain Bt 4D1. Bt RT carried a different cry gene profile than Bt 4D1. Scanning electron microscopy showed the presence of bipyramidal and cuboidal crystals. Phenotypic characterization revealed lytic enzymes that could contribute to Bt pathogenicity. PMID:19693442

  13. Characterization of the Earwig, Doru lineare, as a Predator of Larvae of the Fall Armyworm, Spodoptera frugiperda: A Functional Response Study

    PubMed Central

    Sueldo, Mabel Romero; Bruzzone, Octavio A.; Virla, Eduardo G.

    2010-01-01

    Spodoptera frugiperda Smith (Lepidoptera: Noctuidae) is considered as the most important pest of maize in almost all tropical America. In Argentina, the earwig Doru lineare Eschscholtz (Dermaptera: Forficulidae) has been observed preying on S. frugiperda egg masses in corn crops, but no data about its potential role as a biocontrol agent of this pest have been provided. The predation efficiency of D. lineare on newly emerged S. frugiperda larva was evaluated through a laboratory functional response study. D. lineare showed type II functional response to S. frugiperda larval density, and disc equation estimations of searching efficiency and handling time were (a) = 0.374 and (t) = 182.9 s, respectively. Earwig satiation occurred at 39.4 S. frugiperda larvae. PMID:20575739

  14. Genomic Analysis and Isolation of RNA Polymerase II Dependent Promoters from Spodoptera frugiperda

    PubMed Central

    Bleckmann, Maren; Fritz, Markus H.-Y.; Bhuju, Sabin; Jarek, Michael; Schürig, Margitta; Geffers, Robert; Benes, Vladimir; Besir, Hüseyin; van den Heuvel, Joop

    2015-01-01

    The Baculoviral Expression Vector System (BEVS) is the most commonly used method for high expression of recombinant protein in insect cells. Nevertheless, expression of some target proteins-especially those entering the secretory pathway- provides a severe challenge for the baculovirus infected insect cells, due to the reorganisation of intracellular compounds upon viral infection. Therefore, alternative strategies for recombinant protein production in insect cells like transient plasmid-based expression or stable expression cell lines are becoming more popular. However, the major bottleneck of these systems is the lack of strong endogenous polymerase II dependent promoters, as the strong baculoviral p10 and polH promoters used in BEVS are only functional in presence of the viral transcription machinery during the late phase of infection. In this work we present a draft genome and a transcriptome analysis of Sf21 cells for the identification of the first known endogenous Spodoptera frugiperda promoters. Therefore, putative promoter sequences were identified and selected because of high mRNA level or in analogy to other strong promoters in other eukaryotic organism. The chosen endogenous Sf21 promoters were compared to early viral promoters for their efficiency to trigger eGFP expression using transient plasmid based transfection in a BioLector Microfermentation system. Furthermore, promoter activity was not only shown in Sf21 cells but also in Hi5 cells. The novel endogenous Sf21 promoters were ranked according to their activity and expand the small pool of available promoters for stable insect cell line development and transient plasmid expression in insect cells. The best promoter was used to improve plasmid based transient transfection in insect cells substantially. PMID:26263512

  15. The attraction of Spodoptera frugiperda neonates to cowpea seedlings is mediated by volatiles induced by conspecific herbivory and the elicitor inceptin

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The fall armyworm [FAW; Spodoptera frugiperda (Smith)] often encounters herbivore damage as it disperses as a neonate from an egg mass to an individual feeding site. We investigated orientation responses of dispersing neonates to herbivore damage in cowpea seedings, specifically examining whether ...

  16. Effect of fall armyworm Spodoptera frugiperda (Lepidoptera: Noctuidae) strain and diet on oviposition and development in the parasitoid Euplectrus platyhypenae (Hymenoptera: Eulophidae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Oviposition tendency and subsequent development of the parasitoid wasp Euplectrus platyhypenae Howard was compared using the corn and rice host strains of Spodoptera frugiperda (J. E. Smith) fed corn (Zea mays, ‘Truckers Favorite’) or stargrass (Cynodon nlemfuensis Vanderyst var. nlemfuensis, ‘Floro...

  17. SfDredd, a Novel Initiator Caspase Possessing Activity on Effector Caspase Substrates in Spodoptera frugiperda

    PubMed Central

    Liu, Hao; Wu, Andong; Mei, Long; Liu, Qingzhen

    2016-01-01

    Sf9, a cell line derived from Spodoptera frugiperda, is an ideal model organism for studying insect apoptosis. The first notable study that attempted to identify the apoptotic pathway in Sf9 was performed in 1997 and included the discovery of Sf-caspase-1, an effector caspase of Sf9. However, it was not until 2013 that the first initiator caspase in Sf9, SfDronc, was discovered, and the apoptotic pathway in Sf9 became clearer. In this study, we report another caspase of Sf9, SfDredd. SfDredd is highly similar to insect initiator caspase Dredd homologs. Experimentally, recombinant SfDredd underwent autocleavage and exhibited different efficiencies in cleavage of synthetic caspase substrates. This was attributed to its caspase activity for the predicted active site mutation blocked the above autocleavage and synthetic caspase substrates cleavage activity. SfDredd was capable of not only cleaving Sf-caspase-1 in vitro but also cleaving Sf-caspase-1 and inducing apoptosis when it was co-expressed with Sf-caspase-1 in Sf9 cells. The protein level of SfDredd was increased when Sf9 cells were treated by Actinomycin D, whereas silencing of SfDredd reduced apoptosis and Sf-caspase-1 cleavage induced by Actinomycin D treatment. These results clearly indicate that SfDredd functioned as an apoptotic initiator caspase. Apoptosis induced in Sf9 cells by overexpression of SfDredd alone was not as obvious as that induced by SfDronc alone, and the cleavage sites of Sf-caspase-1 for SfDredd and SfDronc are different. In addition, despite sharing a sequence homology with initiator caspases and possessing weak activity on initiator caspase substrates, SfDredd showed strong activity on effector caspase substrates, making it the only insect caspase reported so far functioning similar to human caspase-2 in this aspect. We believe that the discovery of SfDredd, and its different properties from SfDronc, will improve the understanding of apoptosis pathway in Sf9 cells. PMID:26977926

  18. Nanotoxicological Effects of SiO2 Nanoparticles on Spodoptera frugiperda Sf9 Cells.

    PubMed

    Santo-Orihuela, Pablo L; Foglia, Maria L; Targovnik, Alexandra M; Miranda, Maria V; Desimone, Martin F

    2016-01-01

    The application of silica nanoparticles (NPs) in the biomedical field experienced a great development. The driving forces for these and future developments are the possibility to design NPs with homogeneous size and structure amenable to specific grafting. Moreover, it is possible to tune the characteristics of the NPs to meet the requirements of each specific cell and desired application. Herein, we analyzed the effect of silica NPs of various sizes and surface charge on the viability of Spodoptera frugiperda cells (Sf9 cell line) with the aim of extending the knowledge of possible toxicity of the NPs in the environment and development of new tools for insect control. Moreover, these results will also contribute to develop more effective systems for gene vectors delivery and recombinant proteins expression. Bare silica NPs of 14 nm, 380 nm and 1430 nm as well as amine-modified silica NPs of 131 nm and 448 nm were obtained by the Stöber method. The NPs were characterized by DLS and zeta potential measurements. The cell viability was assessed by the MTT test. It was observed that the 14 nm NPs possess the highest toxic effect. Indeed, after 24 h, the viability of the cells exposed to the lower concentration of NPs (0.12 mg/ml) was about 40% of the value obtained for the control cells not exposed to NPs. Moreover, the exposure to other negative charged NPs also causes a lower activity when compared with the control. Alternatively, lower concentrations of positive charged NPs (i.e.: 0.12 or 0.6 mg/ml) demonstrated to stimulate the proliferation of the cells and higher concentrations (i.e.: 7.2 mg/ml) did not present significant differences with the control. In conclusion, we have demonstrated that the NPs possess an effect that is highly influenced by the size, charge and concentration. Although, silica NPs are being used in the biomedical field, these results contribute to further understanding the risk that could be associated to nanoparticles and how these can be

  19. Ecology, Behavior and Bionomics: First Genotyping of Spodoptera frugiperda (J. E. Smith)(Lepidoptera: Noctuidae) Progeny from Crosses between Bt-Resistant and Bt-Susceptible Populations, and 65-Locus Discrimination of Isofami

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Simple sequence repeat (SSR) markers from Spodoptera frugiperda (J. E. Smith) were analyzed in crosses of this species between Bacillus thuringiensis (Berliner) (Bacillales: Bacillaceae) (Bt) resistant and susceptible populations to determine a possible association between markers and Bt resistance....

  20. Effect of triterpenoids and limonoids isolated from Cabralea canjerana and Carapa guianensis (Meliaceae) against Spodoptera frugiperda (J. E. Smith).

    PubMed

    Sarria, André L F; Soares, Márcio S; Matos, Andréia P; Fernandes, João B; Vieira, Paulo C; da Silva, M Fátima das G F

    2011-01-01

    The activities of two triterpenoids, ocotillone and cabraleadiol, and four limonoids, methyl angolensate, 3-beta-deacetylfissinolide, 7-deacetoxy-7-oxogedunin, and beta-photogedunin, isolated from arillus of Carapa guianensis and fruits and seeds of Cabralea canjerana (Meliaceae), were evaluated against the fall armyworm Spodoptera frugiperda. Gedunin was used as a positive control. 7-Deacetoxy-7-oxogedunin and beta-photogedunin reduced the pupal weight as occurred with gedunin. Cabraleadiol, 3-beta-deacetylfissinolide, and 7-deacetoxy-7-oxogedunin prolonged the larval phase similar to the control (gedunin) of approximately 1.2 days at 50.0 mg kg(-1). The highest insecticidal activity was obtained for beta-photogedunin. PMID:21812341

  1. A Z-linked sterility locus causes sexual abstinence in hybrid females and facilitates speciation in Spodoptera frugiperda.

    PubMed

    Kost, Silvia; Heckel, David G; Yoshido, Atsuo; Marec, František; Groot, Astrid T

    2016-06-01

    In the fall armyworm, Spodoptera frugiperda (Lepidoptera, Noctuidae), two sympatric strains have been recognized that have been termed corn strain (C) and rice strain (R), referring to their most common host plants. Both strains are reproductively isolated via a distinct prezygotic barrier as well as via an intriguing postzygotic phenomenon: when R females have mated with C males, the resulting RC hybrid females exhibit dramatically reduced fertility independent of their mating partner. Here, we demonstrate that the reduced fertility is caused by the fact that these females refrain from mating, that is, females are behaviorally sterile. We identified a Z-chromosomally linked sterility locus that is most likely incompatible with yet to be identified autosomal (or cytoplasmic) factors, leading to the observed sexual abstinence. Within-chromosome mapping revealed the sterility locus to be located in an area of strongly reduced interstrain recombination. PMID:27149933

  2. Resistance to dual-gene Bt maize in Spodoptera frugiperda: selection, inheritance, and cross-resistance to other transgenic events.

    PubMed

    Santos-Amaya, Oscar F; Rodrigues, João V C; Souza, Thadeu C; Tavares, Clébson S; Campos, Silverio O; Guedes, Raul N C; Pereira, Eliseu J G

    2015-01-01

    Transgenic crop "pyramids" producing two or more Bacillus thuringiensis (Bt) toxins active against the same pest are used to delay evolution of resistance in insect pest populations. Laboratory and greenhouse experiments were performed with fall armyworm, Spodoptera frugiperda, to characterize resistance to Bt maize producing Cry1A.105 and Cry2Ab and test some assumptions of the "pyramid" resistance management strategy. Selection of a field-derived strain of S. frugiperda already resistant to Cry1F maize with Cry1A.105 + Cry2Ab maize for ten generations produced resistance that allowed the larvae to colonize and complete the life cycle on these Bt maize plants. Greenhouse experiments revealed that the resistance was completely recessive (Dx = 0), incomplete, autosomal, and without maternal effects or cross-resistance to the Vip3Aa20 toxin produced in other Bt maize events. This profile of resistance supports some of the assumptions of the pyramid strategy for resistance management. However, laboratory experiments with purified Bt toxin and plant leaf tissue showed that resistance to Cry1A.105 + Cry2Ab2 maize further increased resistance to Cry1Fa, which indicates that populations of fall armyworm have high potential for developing resistance to some currently available pyramided maize used against this pest, especially where resistance to Cry1Fa was reported in the field. PMID:26675246

  3. Resistance to dual-gene Bt maize in Spodoptera frugiperda: selection, inheritance, and cross-resistance to other transgenic events

    PubMed Central

    Santos-Amaya, Oscar F.; Rodrigues, João V. C.; Souza, Thadeu C.; Tavares, Clébson S.; Campos, Silverio O.; Guedes, Raul N.C.; Pereira, Eliseu J.G.

    2015-01-01

    Transgenic crop “pyramids” producing two or more Bacillus thuringiensis (Bt) toxins active against the same pest are used to delay evolution of resistance in insect pest populations. Laboratory and greenhouse experiments were performed with fall armyworm, Spodoptera frugiperda, to characterize resistance to Bt maize producing Cry1A.105 and Cry2Ab and test some assumptions of the “pyramid” resistance management strategy. Selection of a field-derived strain of S. frugiperda already resistant to Cry1F maize with Cry1A.105 + Cry2Ab maize for ten generations produced resistance that allowed the larvae to colonize and complete the life cycle on these Bt maize plants. Greenhouse experiments revealed that the resistance was completely recessive (Dx = 0), incomplete, autosomal, and without maternal effects or cross-resistance to the Vip3Aa20 toxin produced in other Bt maize events. This profile of resistance supports some of the assumptions of the pyramid strategy for resistance management. However, laboratory experiments with purified Bt toxin and plant leaf tissue showed that resistance to Cry1A.105 + Cry2Ab2 maize further increased resistance to Cry1Fa, which indicates that populations of fall armyworm have high potential for developing resistance to some currently available pyramided maize used against this pest, especially where resistance to Cry1Fa was reported in the field. PMID:26675246

  4. Effects of Methanolic Extracts of Annona Species on the Development and Reproduction of Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae).

    PubMed

    Freitas, A F; Pereira, F F; Formagio, A S N; Lucchetta, J T; Vieira, M C; Mussury, R M

    2014-10-01

    Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) causes significant losses in corn crops and necessitates the use of alternative control strategies, such as the application of bioinsecticides. We report the effect of methanolic leaf extracts of Annona dioica, Annona cacans, and Annona coriacea on the development and reproduction of S. frugiperda. A quantitative analysis was carried out to determine the total concentration of phenolics, flavonoids, and condensed tannin (CT) in leaf extracts. Corn leaves were immersed in a 1% methanolic leaf extract solution and fed to second instars of S. frugiperda. Leaf disks dipped in the synthetic insecticide Connect® (Bayer CropScience Ltda) composed of a neonicotinoid (imidacloprid) and a pyrethroid (β-cyfluthrin), which are harmful to S. frugiperda, was used as positive control. Distilled water was used as a negative control treatment. The leaf extract of A. coriacea decreased larval survivorship, arrested pupal development, and affected the weight gain of S. frugiperda. A. dioica also affected larval survivorship, but its effects were more pronounced for the adult stage, as fecundity, fertility, egg hatchability, and embryonic development were severely affected. Leaf extracts from A. cacans had no effect on S. frugiperda. The leaf extracts of A. dioica and A. coriacea showed a higher content of flavonoids and phenols, respectively. Our results indicated that both A. dioica and A. coriacea have the potential for development as botanical insecticides. PMID:27193955

  5. Resistance Risk Assessment of Spodoptera frugiperda (Lepidoptera: Noctuidae) and Diatraea saccharalis (Lepidoptera: Crambidae) to Vip3Aa20 Insecticidal Protein Expressed in Corn.

    PubMed

    Bernardi, Oderlei; Bernardi, Daniel; Amado, Douglas; Sousa, Renan S; Fatoretto, Julio; Medeiros, Fernanda C L; Conville, Jared; Burd, Tony; Omoto, Celso

    2015-12-01

    Transgenic Agrisure Viptera 3 corn that expresses Cry1Ab, Vip3Aa20, and EPSPS proteins and Agrisure Viptera expressing Vip3Aa20 are used for control of Spodoptera frugiperda (J.E. Smith) and Diatraea saccharalis (F.) in Brazil. To support a resistance management program, resistance risk assessment studies were conducted to characterize the dose expression of Vip3Aa20 protein and level of control against these species. The Vip3Aa20 expression in Agrisure Viptera 3 and Agrisure Viptera decreased from V6 to V10 stage of growth. However, Vip3Aa20 expression in Agrisure Viptera 3 at V6 and V10 stages was 13- and 16-fold greater than Cry1Ab, respectively. The Vip3Aa20 expression in lyophilized tissue of Agrisure Viptera 3 and Agrisure Viptera diluted 25-fold in an artificial diet caused complete larval mortality of S. frugiperda and D. saccharalis. In contrast, lyophilized tissue of Bt11 at the same dilution does not provide complete mortality of these species. Agrisure Viptera 3 and Agrisure Viptera also caused a high level of mortality against S. frugiperda and D. saccharalis. Moreover, 100% mortality was observed for S. frugiperda larvae (neonates through fifth-instar larvae) when fed in corn with the Vip trait technology. Viptera corn achieves a high level of control against S. frugiperda and D. saccharalis providing a high dose, which is an important determination to support the refuge strategy for an effective resistance management program. PMID:26470366

  6. Selective effects of natural and synthetic insecticides on mortality of Spodoptera frugiperda (Lepidoptera: Noctuidae) and its predator Eriopis connexa (Coleoptera: Coccinellidae).

    PubMed

    Tavares, Wagner S; Costa, Mariana A; Cruz, Ivan; Silveira, Rodrigo D; Serrao, Jose E; Zanuncio, Jose C

    2010-08-01

    Spodoptera frugiperda Smith (Lepidoptera: Noctuidae) is a serious pest of corn in several American countries. It is mainly controlled with synthetic insecticides. The objectives of this study were to evaluate the effects of the natural products, neem oil and pyroligneous extract, and the synthetic insecticide, lufenuron, at 2.50 mL water (0.25%) on the mortality of 2-, 4- and 6-day-old caterpillars of S. frugiperda, and their selectivities against fourth instar larvae of Eriopis connnexa Germar (Coleoptera: Coccinellidae). Four- and 6-day-old S. frugiperda caterpillars showed higher mortality after exposure to neem oil (83.33 +/- 0.83 and 89.58 +/- 0.90%, respectively) and lufenuron (95.83 +/- 0.96 and 85.41 +/- 0.83%), compared to pyroligneous extract (68.75 +/- 0.69 and 31.25 +/- 0.31%). The deleterious effect of pyroligneous extract was higher in 2- (83.33 +/- 0.83% mortality) and 4-day-old (68.75 +/- 0.69%) S. frugiperda caterpillars than in 6-day-old caterpillars (31.25 +/- 0.31%). Larval mortality of the predator E. connexa was lower with neem oil and pyroligneous extract (25.00 +/- 0.33%) than with lufenuron (91.66 +/- 1.22%). Neem oil is thus recommended for control of S. frugiperda because of its high toxicity, combined with its relatively low toxicity to larvae of the natural enemy E. connexa. PMID:20603748

  7. Insect growth regulatory effects of some extracts and sterols from Myrtillocactus geometrizans (Cactaceae) against Spodoptera frugiperda and Tenebrio molitor.

    PubMed

    Céspedes, Carlos L; Salazar, J Rodrigo; Martínez, Mariano; Aranda, Eduardo

    2005-10-01

    A methanol extract from the roots and aerial parts of Myrtillocactus geometrizans (Cactaceae) yielded peniocerol 1, macdougallin 2, and chichipegenin 3. The natural products 1, 2 their mixtures, MeOH and CH(2)Cl(2) extracts showed insecticidal and insect growth regulatory activity against fall armyworm [Spodoptera frugiperda J. E. Smith (Lepidoptera: Noctuidae)], an important insect pest of corn, and [Tenebrio molitor (Coleoptera)], a pest of stored grains in Mexico. The most active compounds were 1, 2, and a mixture (M(2)) of 1 and 2 (6:4). All these extracts, compounds and the mixture had insect growth regulating (IGR) activity between 5.0 and 50.0 ppm and insecticidal effects between 50 and 300 ppm in diets. The extracts were insecticidal to larvae, with lethal doses between 100 and 200 ppm. These compounds appear to have selective effects on the pre-emergence metabolism of Coleoptera, because in all treatments of the larvae of T. molitor, pupation were shortened and this process show precociousness in relation to controls. In contrast to S. frugiperda larvae, onset of pupation was noticeably delayed. Emergence in both cases was drastically diminished. In both pupae and in the few adults that were able to emerge, many deformations were observed. The results of these assays indicated that the compounds were more active than other known natural insect growth inhibitors such as gedunin and methanol extracts of Cedrela salvadorensis and Yucca periculosa. Peniocerol, macdougallin and chichipegenin, as well as mixtures of these substances, may be useful as natural insecticidal agents. PMID:16122768

  8. Acetylcholinesterase and insect growth inhibitory activities of Gutierrezia microcephala on fall armyworm Spodoptera frugiperda J.E. Smith.

    PubMed

    Calderón, J S; Céspedes, C L; Rosas, R; Gómez-Garibay, F; Salazar, J R; Lina, L; Aranda, E; Kubo, I

    2001-01-01

    From the aerial parts of Gutierrezia microcephala (Asteraceae), four oxyflavones were isolated, namely 5,7,2'-trihydroxy-3,6,8,4',5'-pentamethoxyflavone (1); 5,7,4'-trihydroxy-3,6,8-trimethoxyflavone (2); 5,7,2',4'-tetrahydroxy-3,6,8,5'-tetramethoxyflavone (3); 5,2'-dihydroxy-3,6,7,8,4',5'-hexamethoxyflavone (4), and an ent-clerodane, bacchabolivic acid (5). Compounds 1-5, the synthetic methyl ester (6), n-hexane and MeOH extracts were evaluated against the fall armyworm (Spodoptera frugiperda). Gedunin, a known insect growth regulator isolated from Cedrela spp. was used as a positive control. When tested for activity on neonate larvae into the no-choice artificial diet bioassay, flavone (1), clerodane (5), its methyl ester (6), MeOH and n-hexane extracts caused significant larval mortality with MC50 of 3.9, 10.7, 3.46, 7.95 and 7.5 ppm at 7 days, respectively, as well as growth reduction. They also increased the development time of surviving larvae and a significant delay in time to pupation and adult emergence. Acute toxicity against adults of S. frugiperda was also found, 5, 6, gedunin and n-hexane extract had the most potent activity with LD50 value of 6.59, 15.05, 10.78, and 12.79 ppm, respectively. In addition, MeOH, n-hexane extracts, 5, 6 and gedunin caused acetylcholinesterase inhibition with 93.7, 100, 90.2, 62.0 and 100% at 50.0 ppm, respectively; whereas 1-4 exhibited only moderate inhibitory activity. Compounds 1, 5 and 6 showed inhibitory activities comparable with gedunin. These compounds could be responsible of the insect growth inhibitory activity of this plant. PMID:11421454

  9. Helicoverpa zea (Lepidoptera: Noctuidae) and Spodoptera frugiperda (Lepidoptera: Noctuidae) Responses to Sorghum bicolor (Poales: Poaceae) Tissues From Lowered Lignin Lines.

    PubMed

    Dowd, Patrick F; Sattler, Scott E

    2015-01-01

    The presence of lignin within biomass impedes the production of liquid fuels. Plants with altered lignin content and composition are more amenable to lignocellulosic conversion to ethanol and other biofuels but may be more susceptible to insect damage where lignin is an important resistance factor. However, reduced lignin lines of switchgrasses still retained insect resistance in prior studies. Therefore, we hypothesized that sorghum lines with lowered lignin content will also retain insect resistance. Sorghum excised leaves and stalk pith Sorghum bicolor (L.) Moench (Poales: Poaceae) from near isogenic brown midrib (bmr) 6 and 12 mutants lines, which have lowered lignin content and increased lignocellulosic ethanol conversion efficiency, were examined for insect resistance relative to wild-type (normal BTx623). Greenhouse and growth chamber grown plant tissues were fed to first-instar larvae of corn earworms, Helicoverpa zea (Boddie) and fall armyworms Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae), two sorghum major pests. Younger bmr leaves had significantly greater feeding damage in some assays than wild-type leaves, but older bmr6 leaves generally had significantly less damage than wild-type leaves. Caterpillars feeding on the bmr6 leaves often weighed significantly less than those feeding on wild-type leaves, especially in the S. frugiperda assays. Larvae fed the pith from bmr stalks had significantly higher mortality compared with those larvae fed on wild-type pith, which suggested that bmr pith was more toxic. Thus, reducing lignin content or changing subunit composition of bioenergy grasses does not necessarily increase their susceptibility to insects and may result in increased resistance, which would contribute to sustainable production. PMID:25601946

  10. Helicoverpa zea (Lepidoptera: Noctuidae) and Spodoptera frugiperda (Lepidoptera: Noctuidae) Responses to Sorghum bicolor (Poales: Poaceae) Tissues From Lowered Lignin Lines

    PubMed Central

    Dowd, Patrick F.; Sattler, Scott E.

    2015-01-01

    The presence of lignin within biomass impedes the production of liquid fuels. Plants with altered lignin content and composition are more amenable to lignocellulosic conversion to ethanol and other biofuels but may be more susceptible to insect damage where lignin is an important resistance factor. However, reduced lignin lines of switchgrasses still retained insect resistance in prior studies. Therefore, we hypothesized that sorghum lines with lowered lignin content will also retain insect resistance. Sorghum excised leaves and stalk pith Sorghum bicolor (L.) Moench (Poales: Poaceae) from near isogenic brown midrib (bmr) 6 and 12 mutants lines, which have lowered lignin content and increased lignocellulosic ethanol conversion efficiency, were examined for insect resistance relative to wild-type (normal BTx623). Greenhouse and growth chamber grown plant tissues were fed to first-instar larvae of corn earworms, Helicoverpa zea (Boddie) and fall armyworms Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae), two sorghum major pests. Younger bmr leaves had significantly greater feeding damage in some assays than wild-type leaves, but older bmr6 leaves generally had significantly less damage than wild-type leaves. Caterpillars feeding on the bmr6 leaves often weighed significantly less than those feeding on wild-type leaves, especially in the S. frugiperda assays. Larvae fed the pith from bmr stalks had significantly higher mortality compared with those larvae fed on wild-type pith, which suggested that bmr pith was more toxic. Thus, reducing lignin content or changing subunit composition of bioenergy grasses does not necessarily increase their susceptibility to insects and may result in increased resistance, which would contribute to sustainable production. PMID:25601946

  11. Survival and Development of Spodoptera frugiperda and Chrysodeixis includens (Lepidoptera: Noctuidae) on Bt Cotton and Implications for Resistance Management Strategies in Brazil.

    PubMed

    Sorgatto, Rodrigo J; Bernardi, Oderlei; Omoto, Celso

    2015-02-01

    In Brazil, Spodoptera frugiperda (J. E. Smith) and Chrysodeixis includens (Walker) are important cotton pests and target of control of Bollgard II (Cry1Ac/Cry2Ab2) and WideStrike (Cry1Ac/Cry1F) cotton technologies. To subsidize an insect resistance management program, we conducted laboratory studies to evaluate the toxicity of these Bt cotton plants throughout larval development of S. frugiperda and C. includens. In bioassays with leaf disc, the efficacy of both Bt cotton plants against neonates was >80% for S. frugiperda and 100% for C. includens. However, S. frugiperda larvae that survived on Bt cotton had >76% of growth inhibition and stunting. In bioassays with S. frugiperda and C. includens larvae fed on non-Bt near-isoline during different time period (from 3 to 18 d) and then transferred to Bollgard II or WideStrike leaves showed that larval susceptibility decreased as larval age increased. For Bollgard II cotton, in all S. frugiperda instars, there were larvae that reached the pupal and adult stages. In contrast, on WideStrike cotton, a few larvae in fifth and sixth instar completed the biological cycle. For C. includens, some larvae in sixth instar originated adults in both Bt cotton plants. In conclusion, Bollgard II and WideStrike cotton technologies showed high efficacy against neonates of S. frugiperda and C. includens. However, the mortality of these species decreases as larval age increase, allowing insect survival in a possible seed mixture environment and favoring the resistance evolution. PMID:26308821

  12. Harmonia axyridis (Coleoptera: Coccinellidae) Exhibits No Preference between Bt and Non-Bt Maize Fed Spodoptera frugiperda (Lepidoptera: Noctuidae)

    PubMed Central

    Dutra, Carla C.; Koch, Robert L.; Burkness, Eric C.; Meissle, Michael; Romeis, Joerg; Hutchison, William D.; Fernandes, Marcos G.

    2012-01-01

    A recent shift in managing insect resistance to genetically engineered (GE) maize consists of mixing non-GE seed with GE seed known as “refuge in a bag”, which increases the likelihood of predators encountering both prey fed Bt and prey fed non-Bt maize. We therefore conducted laboratory choice-test feeding studies to determine if a predator, Harmonia axyridis, shows any preference between prey fed Bt and non-Bt maize leaves. The prey species was Spodoptera frugiperda, which were fed Bt maize (MON-810), expressing the single Cry1Ab protein, or non-Bt maize. The predators were third instar larvae and female adults of H. axyridis. Individual predators were offered Bt and non-Bt fed prey larvae that had fed for 24, 48 or 72 h. Ten and 15 larvae of each prey type were offered to third instar and adult predators, respectively. Observations of arenas were conducted at 1, 2, 3, 6, 15 and 24 h after the start of the experiment to determine the number and type of prey eaten by each individual predator. Prey larvae that fed on non-Bt leaves were significantly larger than larvae fed Bt leaves. Both predator stages had eaten nearly all the prey by the end of the experiment. However, in all combinations of predator stage and prey age, the number of each prey type consumed did not differ significantly. ELISA measurements confirmed the presence of Cry1Ab in leaf tissue (23–33 µg/g dry weight) and S. frugiperda (2.1–2.2 µg/g), while mean concentrations in H. axyridis were very low (0.01–0.2 µg/g). These results confirm the predatory status of H. axyridis on S. frugiperda and that both H. axyridis adults and larvae show no preference between prey types. The lack of preference between Bt-fed and non-Bt-fed prey should act in favor of insect resistance management strategies using mixtures of GE and non-GE maize seed. PMID:23024772

  13. Baculovirus p35 gene is oppositely regulated by P53 and AP-1 like factors in Spodoptera frugiperda

    SciTech Connect

    Mohareer, Krishnaveni; Sahdev, Sudhir; Hasnain, Seyed E.

    2011-11-04

    Highlights: Black-Right-Pointing-Pointer Baculovirus p35 is regulated by both viral and host factors. Black-Right-Pointing-Pointer Baculovirus p35 is negatively regulated by SfP53-like factor. Black-Right-Pointing-Pointer Baculovirus p35 is positively regulated by SfAP-1-like factor. -- Abstract: Baculovirus p35 belongs to the early class of genes of AcMNPV and requires viral factors like Immediate Early protein-1 for its transcription. To investigate the role of host factors in regulating p35 gene expression, the putative transcription factor binding sites were examined in silico and the role of these factors in influencing the transcription of p35 gene was assessed. We focused our studies on AP-1 and P53-like factors, which are activated under oxidative stress conditions. The AP-1 motif is located at -1401 while P53 motif is at -1912 relative to p35 translation start site. The predicted AP-1 and P53 elements formed specific complexes with Spodoptera frugiperda nuclear extracts. Both AP-1 and P53 motif binding proteins were down regulated as a function of AcMNPV infection in Spodoptera cells. To address the question whether during an oxidative outburst, the p35 transcription is enhanced; we investigated the role of these oxidative stress induced host transcription factors in influencing p35 gene transcription. Reporter assays revealed that AP-1 element enhances the transcription of p35 by a factor of two. Interestingly, P53 element appears to repress the transcription of p35 gene.

  14. Spodoptera frugiperda resistance to oral infection by Autographa californica multiple nucleopolyhedrovirus linked to aberrant occlusion-derived virus binding in the midgut.

    PubMed

    Haas-Stapleton, Eric J; Washburn, Jan O; Volkman, Loy E

    2005-05-01

    Spodoptera frugiperda larvae are highly resistant to oral infection by Autographa californica multiple nucleopolyhedrovirus (AcMNPV) (LD(50), approximately 9200 occlusions), but extremely susceptible to budded virus within the haemocoel (LD(50), <1 p.f.u.). The inability of AcMNPV occlusion-derived virus (ODV) to establish primary infections readily within midgut cells accounts for a major proportion of oral resistance. To determine whether inappropriate binding of AcMNPV ODV to S. frugiperda midgut cells contributes to lack of oral infectivity, the binding and fusion properties of AcMNPV ODV were compared with those of the ODV of a new isolate of Spodoptera frugiperda multiple nucleopolyhedrovirus (SfMNPV) obtained from a field-collected larva (oral LD(50), 12 occlusions). By using a fluorescence-dequenching assay conducted in vivo, it was found that AcMNPV ODV bound to the midgut epithelia of S. frugiperda larvae at approximately 15 % of the level of SfMNPV ODV, but that, once bound, the efficiencies of fusion for the two ODVs were similar: 60 % for AcMNPV and 53 % for SfMNPV. Whilst the difference in binding efficiencies was significant, it could not account entirely for the observed differences in infectivity. Competition experiments, however, revealed that, in S. frugiperda larvae, SfMNPV ODV bound to a midgut cell receptor that was not bound by AcMNPV ODV, indicating that ODV interaction with a specific receptor(s) was necessary for productive infection of midgut columnar epithelial cells. Fusion in the absence of this ligand-receptor interaction did not result in productive infections. PMID:15831946

  15. Imidacloprid affects the functional response of predator Podisus nigrispinus (Dallas) (Heteroptera: Pentatomidae) to strains of Spodoptera frugiperda (J.E. Smith) on Bt cotton.

    PubMed

    Malaquias, J B; Ramalho, F S; Omoto, C; Godoy, W A C; Silveira, R F

    2014-03-01

    Podisus nigrispinus (Dallas) (Heteroptera: Pentatomidae) is one of the most common asopine species in the neotropical region and its occurrence was reported in several countries of South and Central America, as an important biological control agent for many crops. This study was carried out to identify the imidacloprid impacts on the functional response of predator P. nigrispinus fed on Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) strain resistant to lambda-cyhalothrin, on Bt cotton expressing Cry1Ac (Bollgard(®)). Spodoptera frugiperda larvae were used in the following conditions: resistant (1) and susceptible (2) strains to lambda-cyhalothrin fed on Bollgard(®) cotton leaves (DP 404 BG); and resistant (3) and susceptible (4) strains to lambda-cyhalothrin fed on non-genetically modified cotton leaves (cultivar DP4049). The predatory behavior of P. nigrispinus was affected by imidacloprid and the type II asymptotic curve was the one that best described the functional response data. Handling time (T h ) of predator females did not differ among treatments in the presence of imidacloprid. The attack rate did decrease, however, due to an increase in the density of larvae offered. Regardless of the treatment (S. frugiperda strain or cotton cultivar), the predation of P. nigrispinus females on S. frugiperda larvae was significantly lower when exposed to imidacloprid, especially at a density of 16 larvae/predator. The predation behavior of P. nigrispinus on S. frugiperda larvae is affected by the insecticide imidacloprid showing that its applications should be used in cotton crop with caution. PMID:24352830

  16. Oligosaccharide processing in the expression of human plasminogen cDNA by lepidopteran insect (Spodoptera frugiperda) cells

    SciTech Connect

    Davidson, D.J.; Fraser, M.J.; Castellino, F.J. )

    1990-06-12

    A comparison has been made between the Asn{sup 289}-linked oligosaccharide structures of human plasma plasminogen and a recombinant human plasminogen, expressed in lepidopteran insect (Spodoptera frugiperda) cells, after infection of these cells with a recombinant baculovirus containing the entire human plasminogen cDNA. Using anion-exchange liquid chromatography mapping of the oligosaccharide units cleaved from the proteins by glycopeptidase F, compared with elution positions of standard oligosaccharide structures, coupled with monosaccharide compositional analysis, the authors find that the human plasma protein contained only bisialo-biantennary complex-type carbohydrate and asialo-biantennary complex carbohydrate, confirming earlier work published by this laboratory. The glycosylation pattern of the insect cell expressed recombinant human plasminogen showed considerable microheterogeneity, with identifiable high-mannose carbohydrate and truncated high-mannose oligosaccharide. Of major importance, approximately 40% of the oligosaccharide population consisted of complex carbohydrate (bisialo-biantennary), identical in structure with that of the human plasma protein. This the first direct identification of complex carbohydrate in proteins produced in insect cells and demonstrates that trimming and processing of high-mannose carbohydrate into complex-type oligosaccharide can occur. The data indicate that both normal and alternate pathways exist in these cells for incorporation and trimming of high-mannose oligosaccharides and that mannosidases, as well as galactosyl-, hexosaminidasyl-, and sialyltransferases are present, and/or can be induced, in these cells. From these observations, the authors conclude that amino acid sequences and/or protein conformational properties can control oligosaccharide processing events.

  17. Larval and pupal stage of Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae) in sweet and field corn genotypes.

    PubMed

    Santos, L M; Redaelli, L R; Diefenbach, L M G; Efrom, C F S

    2003-11-01

    Spodoptera frugiperda, the fall armyworm, is a very significant polyphagous pest due to the damages it causes, and control difficulties. Lack of information about its impact on sweet corn motivated a comparison of its biology, with respect to the larval and pupal stages, among the genotypes ELISA, BR 400 (sweet corns), and BR PAMPA (field corn). In laboratory conditions (25 +/- 1 masculine C; 70 +/- 10% RH; photophase 12 hours), 35 caterpillars were individualized and fed daily with 3.14 cm(2) sections of corn leaves from the referred-to genotypes, cultivated in plots in the experimental area of the Departament of Fitossanidade, UFRGS, Porto Alegre, RS from October to November 2000. The caterpillars were weighed daily; after each molt, the cephalic capsules were collected and measured (in width), to establish growth rate; pupae were weighed and sexed when 24 hours old. The duration of the larval instars, the pupal sex ratio, and the mortality of larvae and pupae were evaluated. In the first three instars there were no differences registered in capsule width. In the fourth and fifth instars, capsules of caterpillars kept in BR 400 were smaller. The weight of caterpillars and pupae, instar duration and sex ratio did not differ among the genotypes. Pupal phase duration was less in females kept in BR 400. Mortality was greater in the larval phase in ELISA and in the pupal phase in BR PAMPA. PMID:15029373

  18. Resistance to Spodoptera frugiperda (Lepidoptera: Noctuidae) and Euxesta stigmatias (Diptera: Ulidiidae) in sweet corn derived from exogenous and endogenous genetic systems.

    PubMed

    Nuessly, G S; Scully, B T; Hentz, M G; Beiriger, R; Snook, M E; Widstrom, N W

    2007-12-01

    Field trials using Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae) and Euxesta stigmatias Loew (Diptera: Ulidiidae) were conducted to evaluate resistance and potential damage interactions between these two primary corn, Zea mays L., pests against Lepidoptera-resistant corn varieties derived from both endogenous and exogenous sources. The endogenous source of resistance was maysin, a C-glycosyl flavone produced in high concentrations in varieties 'Zapalote Chico 2451' and 'Zapalote Chico sh2'. The exogenous resistance source was the Bacillus thuringiensis (Bt)11 gene that expresses Cry1A(b) insecticidal protein found in 'Attribute GSS-0966'. Damage by the two pests was compared among these resistant varieties and the susceptible 'Primetime'. Single-species tests determined that the Zapalote Chico varieties and GSS-0966 effectively reduced S. frugiperda larval damage compared with Primetime. E. stigmatias larval damage was less in the Zapalote Chico varieties than the other varieties in single-species tests. E. stigmatias damage was greater on S. frugiperda-infested versus S. frugiperda-excluded ears. Ears with S. frugiperda damage to husk, silk and kernels had greater E. stigmatias damage than ears with less S. frugiperda damage. Reversed phase high-performance liquid chromatography analysis of nonpollinated corn silk collected from field plots determined that isoorientin, maysin, and apimaysin plus 3'-methoxymaysin concentrations followed the order Zapalote Chico sh2 > Zapalote Chico 2451 > Attribute GSS-0966 = Primetime. Chlorogenic acid concentrations were greatest in Zapalote Chico 2451. The two high maysin Zapalote Chico varieties did as well against fall armyworm as the Bt-enhanced GSS-0966, and they outperformed GSS-0966 against E. stigmatias. PMID:18232407

  19. Redescription of Campoletis sonorensis (Cameron, 1886) (Hymenoptera, Ichneumonidae, Campopleginae), parasitoid of Spodoptera frugiperda (J. E. Smith, 1797) (Lepidoptera, Noctuidae) in Brazil.

    PubMed

    Camargo, L F; Brito, R A; Penteado-Dias, A M

    2015-11-01

    The fall armyworm Spodoptera frugiperda (Lepidoptera; Noctuidae) is a voracious pest of numerous crops of economic importance throughout the New World. In Brazil, its larvae are attacked by several species of parasitoid wasps, making them potential candidate as biological control agents against this pest. A survey of the parasitoid fauna on S. frugiperda in maize crops throughout Brazil reveals two species of Campoletis, which are morphologicaly very similar species. In this paper we combine these data with pictures from the type material of C. sonorensis and C. flavicincta, as well as their descriptions to provide a redescription to Campoletis sonorensis (Cameron, 1886) using for this both morphological characters and DNA Barcoding (Hebert et al., 2003) information, in an attempt to help with the correct identification of the taxa to improve biological control studies. PMID:26675917

  20. Proteomics of the 26S proteasome in Spodoptera frugiperda cells infected with the nucleopolyhedrovirus, AcMNPV.

    PubMed

    Lyupina, Yulia V; Zatsepina, Olga G; Serebryakova, Marina V; Erokhov, Pavel A; Abaturova, Svetlana B; Kravchuk, Oksana I; Orlova, Olga V; Beljelarskaya, Svetlana N; Lavrov, Andrey I; Sokolova, Olga S; Mikhailov, Victor S

    2016-06-01

    Baculoviruses are large DNA viruses that infect insect species such as Lepidoptera and are used in biotechnology for protein production and in agriculture as insecticides against crop pests. Baculoviruses require activity of host proteasomes for efficient reproduction, but how they control the cellular proteome and interact with the ubiquitin proteasome system (UPS) of infected cells remains unknown. In this report, we analyzed possible changes in the subunit composition of 26S proteasomes of the fall armyworm, Spodoptera frugiperda (Sf9), cells in the course of infection with the Autographa californica multiple nucleopolyhedrovirus (AcMNPV). 26S proteasomes were purified from Sf9 cells by an immune affinity method and subjected to 2D gel electrophoresis followed by MALDI-TOF mass spectrometry and Mascot search in bioinformatics databases. A total of 34 homologues of 26S proteasome subunits of eukaryotic species were identified including 14 subunits of the 20S core particle (7 α and 7 β subunits) and 20 subunits of the 19S regulatory particle (RP). The RP contained homologues of 11 of RPN-type and 6 of RPT-type subunits, 2 deubiquitinating enzymes (UCH-14/UBP6 and UCH-L5/UCH37), and thioredoxin. Similar 2D-gel maps of 26S proteasomes purified from uninfected and AcMNPV-infected cells at 48hpi confirmed the structural integrity of the 26S proteasome in insect cells during baculovirus infection. However, subtle changes in minor forms of some proteasome subunits were detected. A portion of the α5(zeta) cellular pool that presumably was not associated with the proteasome underwent partial proteolysis at a late stage in infection. PMID:26945516

  1. Comparative proteomics analysis of apoptotic Spodoptera frugiperda cells during p35 knockout Autographa californica multiple nucleopolyhedrovirus infection.

    PubMed

    Yu, Qian; Xiong, Youhua; Liu, Jianliang; Wang, Qin; Qiu, Yuanxin; Wen, Dongling

    2016-06-01

    Infection with Autographa californica multiple nucleopolyhedrovirus (AcMNPV) mutants lacking a functional p35 gene can induce host cell apoptosis, which provides the possibility to use the potential of these viruses in the biological control of pest insects. Nonetheless, the proteomics or the protein changes of Spodoptera frugiperda (Sf9) cells infected with p35 knockout AcMNPV have not yet been studied. To further improve the use of AcMNPV, we set out to analyze the protein composition and protein changes of Sf9 cells of different infection stages by isobaric tag for relative and absolute quantification (iTRAQ) techniques. A total of 4004 sf9 proteins were identified by iTRAQ. After comparation of the significantly expressed 483 proteins from p35koAcMNPV-infected Sf9 cells and the significantly expressed 413 proteins from wtAcMNPV-infected Sf9 cells, we found that 226 proteins were specific to p35koAcMNPV-infected Sf9 cells. The 226 proteins were categorized according to GO classification for insects and were categorized into: biological processes, molecular functions and cellular components. Of interest, the most up-regulated proteins related to Epstein-Barr virus infection, RNA transport, Calcium signaling pathway, cGMP-PKG signaling pathway, oxidative phosphorylation and N-Glycan biosynthesis. Determination of the protein changes in p35 knockout AcMNPV-infected Sf9 cells would facilitate the better use of this virus-host cell interaction in pest insect control and other related fields. PMID:26922645

  2. Deletion Genotypes Reduce Occlusion Body Potency but Increase Occlusion Body Production in a Colombian Spodoptera frugiperda Nucleopolyhedrovirus Population

    PubMed Central

    Barrera, Gloria; Williams, Trevor; Villamizar, Laura; Caballero, Primitivo; Simón, Oihane

    2013-01-01

    A Colombian field isolate (SfCOL-wt) of Spodoptera frugiperda multiple nucleopolyhedrovirus (SfMNPV) is a mixture of different genotypes. To evaluate the insecticidal properties of the different genotypic variants, 83 plaque purified virus were characterized. Ten distinct genotypes were identified (named A through J). SfCOL-A was the most prevalent (71±2%; mean ± SE) showing a PstI restriction profile indistinguishable to that of SfCOL-wt. The remaining nine genotypes presented genomic deletions of 3.8 - 21.8 Kb located mainly between nucleotides 11,436 and 33,883 in the reference genome SfMNPV-B, affecting the region between open reading frames (ORFs) sf20 and sf33. The insecticidal activity of each genotype from SfCOL-wt and several mixtures of genotypes was compared to that of SfCOL-wt. The potency of SfCOL-A occlusion bodies (OBs) was 4.4-fold higher than SfCOL-wt OBs, whereas the speed of kill of SfCOL-A was similar to that of SfCOL-wt. Deletion genotype OBs were similarly or less potent than SfCOL-wt but six deletion genotypes were faster killing than SfCOL-wt. The potency of genotype mixtures co-occluded within OBs were consistently reduced in two-genotype mixtures involving equal proportions of SfCOL-A and one of three deletion genotypes (SfCOL-C, -D or -F). Speed of kill and OB production were improved only when the certain genotype mixtures were co-occluded, although OB production was higher in the SfCOL-wt isolate than in any of the component genotypes, or mixtures thereof. Deleted genotypes reduced OB potency but increased OB production of the SfCOL-wt population, which is structured to maximize the production of OBs in each infected host. PMID:24116220

  3. Comparative Effectiveness of Potential Elicitors of Plant Resistance against Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae) in Four Crop Plants

    PubMed Central

    Gordy, John W.; Leonard, B. Rogers; Blouin, David; Davis, Jeffrey A.; Stout, Michael J.

    2015-01-01

    Feeding by insect herbivores activates plant signaling pathways, resulting in the enhanced production of secondary metabolites and other resistance-related traits by injured plants. These traits can reduce insect fitness, deter feeding, and attract beneficial insects. Organic and inorganic chemicals applied as a foliar spray, seed treatment, or soil drench can activate these plant responses. Azelaic acid (AA), benzothiadiazole (BTH), gibberellic acid (GA), harpin, and jasmonic acid (JA) are thought to directly mediate plant responses to pathogens and herbivores or to mimic compounds that do. The effects of these potential elicitors on the induction of plant defenses were determined by measuring the weight gains of fall armyworm, Spodoptera frugiperda (J. E. Smith) (FAW) (Lepidoptera: Noctuidae) larvae on four crop plants, cotton, corn, rice, and soybean, treated with the compounds under greenhouse conditions. Treatment with JA consistently reduced growth of FAW reared on treated cotton and soybean. In contrast, FAW fed BTH- and harpin-treated cotton and soybean tissue gained more weight than those fed control leaf tissue, consistent with negative crosstalk between the salicylic acid and JA signaling pathways. No induction or inconsistent induction of resistance was observed in corn and rice. Follow-up experiments showed that the co-application of adjuvants with JA failed to increase the effectiveness of induction by JA and that soybean looper [Chrysodeixis includens (Walker)], a relative specialist on legumes, was less affected by JA-induced responses in soybean than was the polyphagous FAW. Overall, the results of these experiments demonstrate that the effectiveness of elicitors as a management tactic will depend strongly on the identities of the crop, the pest, and the elicitor involved. PMID:26332833

  4. Performance and cross-crop resistance of Cry1F-maize selected Spodoptera frugiperda on transgenic Bt cotton: implications for resistance management

    PubMed Central

    Yang, Fei; Kerns, David L.; Brown, Sebe; Kurtz, Ryan; Dennehy, Tim; Braxton, Bo; Head, Graham; Huang, Fangneng

    2016-01-01

    Transgenic crops producing Bacillus thuringiensis (Bt) proteins have become a primary tool in pest management. Due to the intensive use of Bt crops, resistance of the fall armyworm, Spodoptera frugiperda, to Cry1F maize has occurred in Puerto Rico, Brazil, and some areas of the southeastern U.S. The sustainability of Bt crops faces a great challenge because the Cry1F-maize resistant S. frugiperda may also infest other Bt crops in multiple cropping ecosystems. Here we examined the survival and plant injury of a S. frugiperda population selected with Cry1F maize on three single-gene and five pyramided Bt cotton products. Larvae of Cry1F-susceptible (SS), -heterozygous (RS), and -resistant (RR) genotypes of S. frugiperda were all susceptible to the pyramided cotton containing Cry1Ac/Cry2Ab, Cry1Ac/Cry1F/Vip3A, Cry1Ab/Cry2Ae, or Cry1Ab/Cry2Ae/Vip3A, and the single-gene Cry2Ae cotton. Pyramided cotton containing Cry1Ac/Cry1F was effective against SS and RS, but not for RR. These findings show that the Cry1F-maize selected S. frugiperda can cause cross-crop resistance to other Bt crops expressing similar insecticidal proteins. Resistance management and pest management programs that utilize diversify mortality factors must be implemented to ensure the sustainability of Bt crops. This is especially important in areas where resistance to single-gene Bt crops is already widespread. PMID:27301612

  5. Performance and cross-crop resistance of Cry1F-maize selected Spodoptera frugiperda on transgenic Bt cotton: implications for resistance management.

    PubMed

    Yang, Fei; Kerns, David L; Brown, Sebe; Kurtz, Ryan; Dennehy, Tim; Braxton, Bo; Head, Graham; Huang, Fangneng

    2016-01-01

    Transgenic crops producing Bacillus thuringiensis (Bt) proteins have become a primary tool in pest management. Due to the intensive use of Bt crops, resistance of the fall armyworm, Spodoptera frugiperda, to Cry1F maize has occurred in Puerto Rico, Brazil, and some areas of the southeastern U.S. The sustainability of Bt crops faces a great challenge because the Cry1F-maize resistant S. frugiperda may also infest other Bt crops in multiple cropping ecosystems. Here we examined the survival and plant injury of a S. frugiperda population selected with Cry1F maize on three single-gene and five pyramided Bt cotton products. Larvae of Cry1F-susceptible (SS), -heterozygous (RS), and -resistant (RR) genotypes of S. frugiperda were all susceptible to the pyramided cotton containing Cry1Ac/Cry2Ab, Cry1Ac/Cry1F/Vip3A, Cry1Ab/Cry2Ae, or Cry1Ab/Cry2Ae/Vip3A, and the single-gene Cry2Ae cotton. Pyramided cotton containing Cry1Ac/Cry1F was effective against SS and RS, but not for RR. These findings show that the Cry1F-maize selected S. frugiperda can cause cross-crop resistance to other Bt crops expressing similar insecticidal proteins. Resistance management and pest management programs that utilize diversify mortality factors must be implemented to ensure the sustainability of Bt crops. This is especially important in areas where resistance to single-gene Bt crops is already widespread. PMID:27301612

  6. Bioactivity of Piper hispidinervum (Piperales: Piperaceae) and Syzygium aromaticum (Myrtales: Myrtaceae) oils, with or without formulated Bta on the biology and immunology of Spodoptera frugiperda (Lepidoptera: Noctuidae).

    PubMed

    Cruz, G S; Wanderley-Teixeira, V; Oliveira, J V; Correia, A A; Breda, M O; Alves, T J S; Cunha, F M; Teixeira, A A C; Dutra, K A; Navarro, D M A F

    2014-02-01

    The combination of essential oils and Bacillus thuringiensis Berliner may represent an interesting control strategy. Thus, the study tested the following hypothesis: the combination of long pepper oil (Piper hispidinervum L.) and clove (Syzygium aromaticum L.) oils in two concentrations with Xentari WG (Bta) yields a more effective control of Spodoptera frugiperda (JE Smith) (Lepidoptera: Noctuidae) affecting biological and reproductive parameters and leading to changes in the levels of phenoloxidase and nitric oxide in the hemolymph of the pest. The results demonstrate that only long pepper oil, at the highest concentration with Xentari WG (Bta), promotes reduced larval survival. However, both oils with or without the insecticide interfere in the biology and humoral immunity of S.frugiperda. All treatments caused a decrease in the amount of eggs, except for the clove oil at both concentrations without Bta. Therefore, the use of these oils is a promising alternative for the integrated management of S. frugiperda; however, its association with Bta demonstrated no significant increase in their efficiency. PMID:24665696

  7. α-Amanitin-Resistant Viral RNA Synthesis in Nuclei Isolated from Nuclear Polyhedrosis Virus-Infected Heliothis zea Larvae and Spodoptera frugiperda Cells

    PubMed Central

    Grula, Marjori A.; Buller, Patricia L.; Weaver, Robert F.

    1981-01-01

    [3H]RNA was synthesized in nuclei isolated at various times postinfection from the fat bodies of Heliothis zea larvae infected with H. zea nuclear polyhedrosis virus and from cultured Spodoptera frugiperda cells infected with Autographa californica nuclear polyhedrosis virus. To detect virus-specific RNA synthesis, the [3H]RNA was hybridized to denatured viral DNA immobilized on nitrocellulose filters. Nuclear polyhedrosis virus-specific RNA synthesis in the infected nuclei isolated from H. zea larval fat bodies and S. frugiperda cells was only inhibited 20 to 25% by concentrations of α-amanitin sufficient to inhibit the host RNA polymerase II. In addition, a productive nuclear polyhedrosis virus infection was obtained in S. frugiperda cells grown in the presence of an α-amanitin concentration that inhibited 90% of the cellular RNA polymerase II activity. The cellular RNA polymerase II enzyme remained sensitive to α-amanitin during infection, and there was no evidence that a virus-coded, α-amanitin-resistant enzyme was synthesized after the onset of infection. The data suggest that the bulk of nuclear polyhedrosis virus-specific RNA synthesis in isolated nuclei is transcribed by an enzyme other than the host RNA polymerase II. PMID:16789208

  8. Evidence of Field-Evolved Resistance of Spodoptera frugiperda to Bt Corn Expressing Cry1F in Brazil That Is Still Sensitive to Modified Bt Toxins

    PubMed Central

    Monnerat, Rose; Martins, Erica; Macedo, Cristina; Queiroz, Paulo; Praça, Lilian; Soares, Carlos Marcelo; Moreira, Helio; Grisi, Isabella; Silva, Joseane; Soberon, Mario; Bravo, Alejandra

    2015-01-01

    Brazil ranked second only to the United States in hectares planted to genetically modified crops in 2013. Recently corn producers in the Cerrado region reported that the control of Spodoptera frugiperda with Bt corn expressing Cry1Fa has decreased, forcing them to use chemicals to reduce the damage caused by this insect pest. A colony of S. frugiperda was established from individuals collected in 2013 from Cry1Fa corn plants (SfBt) in Brazil and shown to have at least more than ten-fold higher resistance levels compared with a susceptible colony (Sflab). Laboratory assays on corn leaves showed that in contrast to SfLab population, the SfBt larvae were able to survive by feeding on Cry1Fa corn leaves. The SfBt population was maintained without selection for eight generations and shown to maintain high levels of resistance to Cry1Fa toxin. SfBt showed higher cross-resistance to Cry1Aa than to Cry1Ab or Cry1Ac toxins. As previously reported, Cry1A toxins competed the binding of Cry1Fa to brush border membrane vesicles (BBMV) from SfLab insects, explaining cross-resistance to Cry1A toxins. In contrast Cry2A toxins did not compete Cry1Fa binding to SfLab-BBMV and no cross-resistance to Cry2A was observed, although Cry2A toxins show low toxicity to S. frugiperda. Bioassays with Cry1AbMod and Cry1AcMod show that they are highly active against both the SfLab and the SfBt populations. The bioassay data reported here show that insects collected from Cry1Fa corn in the Cerrado region were resistant to Cry1Fa suggesting that resistance contributed to field failures of Cry1Fa corn to control S. frugiperda. PMID:25830928

  9. Baseline susceptibility and monitoring of Brazilian populations of Spodoptera frugiperda (Lepidoptera: Noctuidae) and Diatraea saccharalis (Lepidoptera: Crambidae) to Vip3Aa20 insecticidal protein.

    PubMed

    Bernardi, Oderlei; Amado, Douglas; Sousa, Renan S; Segatti, Fabiana; Fatoretto, Julio; Burd, Anthony D; Omoto, Celso

    2014-04-01

    The genetically modified maize expressing Vip3Aa20 insecticidal protein from Bacillus thuringiensis Berliner is abiotechnological option for the control of Spodoptera frugiperda (J.E. Smith) and Diatraea saccharalis (F.) in Brazil. To support an Insect Resistance Management program, we conducted studies of baseline susceptibility and monitoring of Brazilian populations of S. frugiperda and D. saccharalis to the Vip3Aa20 insecticidal protein. Neonates were exposed to Vip3Aa20 applied on artificial diet surface. Mortality and growth inhibition were assessed after 7 d. All populations were susceptible to Vip3Aa20. The LC50 ranged from 92.38 to 611.65 ng Vip3Aa20/cm2 for 16 populations of S. frugiperda (6.6-fold variation), and between 61.18 and 367.86 ng Vip3Aa20/cm2 for 6 populations of D. saccharalis (sixfold variation). The EC50 ranged from 21.76 to 70.09 and 48.65 to 163.60 ng Vip3Aa20/cm2 for S. frugiperda and D. saccharalis, respectively. There was a low interpopulation variation in susceptibility to Vip3Aa20, which represents the natural geographic variation in the response, and not the variation caused by previous exposure to selection pressure. For these two pests, the diagnostic concentrations of 2,000 and 3,600 ng of Vip3Aa20/cm2 caused high mortality. These diagnostic concentrations will be used in resistance monitoring programs in Brazil. PMID:24772561

  10. Evidence of field-evolved resistance of Spodoptera frugiperda to Bt corn expressing Cry1F in Brazil that is still sensitive to modified Bt toxins.

    PubMed

    Monnerat, Rose; Martins, Erica; Macedo, Cristina; Queiroz, Paulo; Praça, Lilian; Soares, Carlos Marcelo; Moreira, Helio; Grisi, Isabella; Silva, Joseane; Soberon, Mario; Bravo, Alejandra

    2015-01-01

    Brazil ranked second only to the United States in hectares planted to genetically modified crops in 2013. Recently corn producers in the Cerrado region reported that the control of Spodoptera frugiperda with Bt corn expressing Cry1Fa has decreased, forcing them to use chemicals to reduce the damage caused by this insect pest. A colony of S. frugiperda was established from individuals collected in 2013 from Cry1Fa corn plants (SfBt) in Brazil and shown to have at least more than ten-fold higher resistance levels compared with a susceptible colony (Sflab). Laboratory assays on corn leaves showed that in contrast to SfLab population, the SfBt larvae were able to survive by feeding on Cry1Fa corn leaves. The SfBt population was maintained without selection for eight generations and shown to maintain high levels of resistance to Cry1Fa toxin. SfBt showed higher cross-resistance to Cry1Aa than to Cry1Ab or Cry1Ac toxins. As previously reported, Cry1A toxins competed the binding of Cry1Fa to brush border membrane vesicles (BBMV) from SfLab insects, explaining cross-resistance to Cry1A toxins. In contrast Cry2A toxins did not compete Cry1Fa binding to SfLab-BBMV and no cross-resistance to Cry2A was observed, although Cry2A toxins show low toxicity to S. frugiperda. Bioassays with Cry1AbMod and Cry1AcMod show that they are highly active against both the SfLab and the SfBt populations. The bioassay data reported here show that insects collected from Cry1Fa corn in the Cerrado region were resistant to Cry1Fa suggesting that resistance contributed to field failures of Cry1Fa corn to control S. frugiperda. PMID:25830928

  11. Cross-Resistance between Cry1 Proteins in Fall Armyworm (Spodoptera frugiperda) May Affect the Durability of Current Pyramided Bt Maize Hybrids in Brazil

    PubMed Central

    Bernardi, Daniel; Salmeron, Eloisa; Horikoshi, Renato Jun; Bernardi, Oderlei; Dourado, Patrick Marques; Carvalho, Renato Assis; Martinelli, Samuel; Head, Graham P.; Omoto, Celso

    2015-01-01

    Genetically modified plants expressing insecticidal proteins from Bacillus thuringiensis (Bt) offer valuable options for managing insect pests with considerable environmental and economic benefits. Despite the benefits provided by Bt crops, the continuous expression of these insecticidal proteins imposes strong selection for resistance in target pest populations. Bt maize (Zea mays) hybrids have been successful in controlling fall armyworm (Spodoptera frugiperda), the main maize pest in Brazil since 2008; however, field-evolved resistance to the protein Cry1F has recently been reported. Therefore it is important to assess the possibility of cross-resistance between Cry1F and other Cry proteins expressed in Bt maize hybrids. In this study, an F2 screen followed by subsequent selection on MON 89034 maize was used to select an S. frugiperda strain (RR) able to survive on the Bt maize event MON 89034, which expresses the Cry1A.105 and Cry2Ab2 proteins. Field-collected insects from maize expressing the Cry1F protein (event TC1507) represented most of the positive (resistance allele-containing) (iso)families found. The RR strain showed high levels of resistance to Cry1F, which apparently also conferred high levels of cross resistance to Cry1A.105 and Cry1Ab, but had only low-level (10-fold) resistance to Cry2Ab2. Life history studies to investigate fitness costs associated with the resistance in RR strain revealed only small reductions in reproductive rate when compared to susceptible and heterozygous strains, but the RR strain produced 32.2% and 28.4% fewer females from each female relative to the SS and RS (pooled) strains, respectively. Consistent with the lack of significant resistance to Cry2Ab2, MON 89034 maize in combination with appropriate management practices continues to provide effective control of S. frugiperda in Brazil. Nevertheless, the occurrence of Cry1F resistance in S. frugiperda across Brazil, and the cross-resistance to Cry1Ab and Cry1A.105

  12. Cross-Resistance between Cry1 Proteins in Fall Armyworm (Spodoptera frugiperda) May Affect the Durability of Current Pyramided Bt Maize Hybrids in Brazil.

    PubMed

    Bernardi, Daniel; Salmeron, Eloisa; Horikoshi, Renato Jun; Bernardi, Oderlei; Dourado, Patrick Marques; Carvalho, Renato Assis; Martinelli, Samuel; Head, Graham P; Omoto, Celso

    2015-01-01

    Genetically modified plants expressing insecticidal proteins from Bacillus thuringiensis (Bt) offer valuable options for managing insect pests with considerable environmental and economic benefits. Despite the benefits provided by Bt crops, the continuous expression of these insecticidal proteins imposes strong selection for resistance in target pest populations. Bt maize (Zea mays) hybrids have been successful in controlling fall armyworm (Spodoptera frugiperda), the main maize pest in Brazil since 2008; however, field-evolved resistance to the protein Cry1F has recently been reported. Therefore it is important to assess the possibility of cross-resistance between Cry1F and other Cry proteins expressed in Bt maize hybrids. In this study, an F2 screen followed by subsequent selection on MON 89034 maize was used to select an S. frugiperda strain (RR) able to survive on the Bt maize event MON 89034, which expresses the Cry1A.105 and Cry2Ab2 proteins. Field-collected insects from maize expressing the Cry1F protein (event TC1507) represented most of the positive (resistance allele-containing) (iso)families found. The RR strain showed high levels of resistance to Cry1F, which apparently also conferred high levels of cross resistance to Cry1A.105 and Cry1Ab, but had only low-level (10-fold) resistance to Cry2Ab2. Life history studies to investigate fitness costs associated with the resistance in RR strain revealed only small reductions in reproductive rate when compared to susceptible and heterozygous strains, but the RR strain produced 32.2% and 28.4% fewer females from each female relative to the SS and RS (pooled) strains, respectively. Consistent with the lack of significant resistance to Cry2Ab2, MON 89034 maize in combination with appropriate management practices continues to provide effective control of S. frugiperda in Brazil. Nevertheless, the occurrence of Cry1F resistance in S. frugiperda across Brazil, and the cross-resistance to Cry1Ab and Cry1A.105

  13. In Vivo and In Vitro Binding of Vip3Aa to Spodoptera frugiperda Midgut and Characterization of Binding Sites by 125I Radiolabeling

    PubMed Central

    Chakroun, Maissa

    2014-01-01

    Bacillus thuringiensis vegetative insecticidal proteins (Vip3A) have been recently introduced in important crops as a strategy to delay the emerging resistance to the existing Cry toxins. The mode of action of Vip3A proteins has been studied in Spodoptera frugiperda with the aim of characterizing their binding to the insect midgut. Immunofluorescence histological localization of Vip3Aa in the midgut of intoxicated larvae showed that Vip3Aa bound to the brush border membrane along the entire apical surface. The presence of fluorescence in the cytoplasm of epithelial cells seems to suggest internalization of Vip3Aa or a fragment of it. Successful radiolabeling and optimization of the binding protocol for the 125I-Vip3Aa to S. frugiperda brush border membrane vesicles (BBMV) allowed the determination of binding parameters of Vip3A proteins for the first time. Heterologous competition using Vip3Ad, Vip3Ae, and Vip3Af as competitor proteins showed that they share the same binding site with Vip3Aa. In contrast, when using Cry1Ab and Cry1Ac as competitors, no competitive binding was observed, which makes them appropriate candidates to be used in combination with Vip3A proteins in transgenic crops. PMID:25002420

  14. Toxicity of Porella chilensis sesqui- and diterpenoids against larvae of the corn pest Spodoptera frugiperda (J.E. Smith) (Lepidotera: Noctuidae).

    PubMed

    Corzo, F L; Gilabert, M; Alcaide, M F; Bardón, A

    2012-10-01

    Porella, the largest genus of the family Porellaceae (Hepaticae) is widespread in the tropical and subtropical regions of South America. Most Porella species are rich sources of sesqui- and diterpenoids, many of which show interesting biological activities. Secondary metabolites produced by plants can interact with insects and act as antifeedants and growth regulators affecting hormone and nervous systems as well as stomach and muscle tissues. A previous chemical investigation of a Patagonian collection of Porella chilensis yielded sesqui- and diterpenoids that were now evaluated for their effects against Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae), a serious pest affecting corn crops mainly in the Americas. Four pinguisanes (1-4), three fusicoccanes (5-7), and one aromadendrane (8) from P. chilensis displayed larvicidal activity against S. frugiperda when incorporated to the larval diet at 100 and 200 μg/g of diet with a significant decrease in the larval growing rate. The observed effects were in part produced by severe alterations of the epithelial cells of the midgut as indicated by our histological studies. PMID:23950093

  15. Population genetic structure of two primary parasitoids of Spodoptera frugiperda (Lepidoptera), Chelonus insularis and Campoletis sonorensis (Hymenoptera): to what extent is the host plant important?

    PubMed

    Jourdie, Violaine; Alvarez, Nadir; Molina-Ochoa, Jaime; Williams, Trevor; Bergvinson, David; Benrey, Betty; Turlings, Ted C J; Franck, Pierre

    2010-05-01

    Plant chemistry can strongly influence interactions between herbivores and their natural enemies, either by providing volatile compounds that serve as foraging cues for parasitoids or predators, or by affecting the quality of herbivores as hosts or prey. Through these effects plants may influence parasitoid population genetic structure. We tested for a possible specialization on specific crop plants in Chelonus insularis and Campoletis sonorensis, two primary parasitoids of the fall armyworm, Spodoptera frugiperda. Throughout Mexico, S. frugiperda larvae were collected from their main host plants, maize and sorghum and parasitoids that emerged from the larvae were used for subsequent comparison by molecular analysis. Genetic variation at eight and 11 microsatellites were respectively assayed for C. insularis and C. sonorensis to examine isolation by distance, host plant and regional effects. Kinship analyses were also performed to assess female migration among host-plants. The analyses showed considerable within population variation and revealed a significant regional effect. No effect of host plant on population structure of either of the two parasitoid species was found. Isolation by distance was observed at the individual level, but not at the population level. Kinship analyses revealed significantly more genetically related--or kin--individuals on the same plant species than on different plant species, suggesting that locally, mothers preferentially stay on the same plant species. Although the standard population genetics parameters showed no effect of plant species on population structure, the kinship analyses revealed that mothers exhibit plant species fidelity, which may speed up divergence if adaptation were to occur. PMID:20406384

  16. Biochemical parameters of Spodoptera frugiperda (J. E. Smith) treated with citronella oil (Cymbopogon winterianus Jowitt ex Bor) and its influence on reproduction.

    PubMed

    Silva, Cristiane Thalita Dos Santos; Wanderley-Teixeira, Valéria; Cunha, Franklin Magliano da; Oliveira, José Vargas de; Dutra, Kamilla de Andrade; Navarro, Daniela Maria do Amaral Ferraz; Teixeira, Álvaro Aguiar Coelho

    2016-05-01

    Spodoptera frugiperda is the principal corn pest in Brazil. Searches for new control methods that minimize the adverse effects of synthetic insecticides have initiated a resurgence of the use of botanical insecticides. Citronella oil (a product of Cymbopogon winterianus) is an effective repellent and insecticide. Thus, biochemical profile changes in oil-treated larvae and its influence on reproduction were assessed. Corn leaves dipped in a 50mg/mL concentration were offered to third instar larvae for 24h and assessed in sixth instar to estimate protein, lipid, sugar, and glycogen levels. Adult testes and ovarioles were collected for histological and histochemical analysis 24h after emergence. Number of eggs and hatching rate were also measured. Oil-treated larvae showed an increase in glycogen and a decrease in protein, lipid, and totals sugar content. Control testes exhibited connective tissue lining and cysts with abundant spermatozoids. However, intense peripheral vacuolation and neutral carbohydrates reduction occurred in oil-treated individuals. Control ovarioles showed normal morphologic characteristics. On the other hand, oil-treatment ovarioles showed follicular cell stratification and removal, reduced nurse cell development, reduced yolk quantity, a thinner conjunctiva sheath, and a reduction in proteins and neutral carbohydrates. Eggs derived from oil-treated pairs were unviable. Therefore, sub-lethal doses of citronella oil alters the biochemical profile of S. frugiperda larvae, causing damage to their reproductive histophysiology and results in diminished reproduction or reproductive failure. PMID:27012436

  17. F2 screen for resistance to Bacillus thuringiensis Cry2Ab2-maize in field populations of Spodoptera frugiperda (Lepidoptera: Noctuidae) from the southern United States.

    PubMed

    Niu, Ying; Qureshi, Jawwad A; Ni, Xinzhi; Head, Graham P; Price, Paula A; Meagher, Robert L; Kerns, David; Levy, Ronnie; Yang, Xiangbing; Huang, Fangneng

    2016-07-01

    The fall armyworm, Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae), is a target pest of transgenic maize and cotton expressing Bacillus thuringiensis (Bt) proteins in both North and South America. In 2013 and 2014, a total of 215 F2 two-parent families of S. frugiperda were established using single-pair mating of field individuals collected from seven locations in four states of the southern U.S.: Texas, Louisiana, Georgia, and Florida. The objective of the investigation was to detect resistance alleles in field populations to Cry2Ab2, a common Bt protein produced in transgenic maize and cotton. For each F2 family, 128 F2 neonates were screened on leaf tissue of Cry2Ab2 maize plants in the laboratory. A conservative estimate of the frequency of major Cry2Ab2 resistance alleles in S. frugiperda from the four states was 0.0023 with a 95% credibility interval of 0.0003-0.0064. In addition, six families were considered to likely possess minor resistance alleles at a frequency of 0.0082 with a 95% credibility interval of 0.0033-0.0152. One F2 family from Georgia (GA-15) was confirmed to possess a major resistance allele to the Cry2Ab2 protein. Larvae from this family survived well on whole maize plants expressing Cry2Ab2 protein and demonstrated a significant level (>15-fold) of resistance when fed with the same protein incorporated in a meridic diet. The detection of the major resistance allele along with the relatively abundant minor resistance alleles revealed in this study may have important implications for resistance management. PMID:27311896

  18. First report of Dolichozele koebelei Viereck, 1911 (Hymenoptera: Braconidae) on larvae of Spodoptera frugiperda (J. E. Smith, 1797) (Lepidoptera: Noctuidae) in maize (Zea mays L.) under different cropping systems.

    PubMed

    Silva, R B; Cruz, I; Penteado-Dias, A M

    2014-08-01

    In the context of the modern agriculture, pest control is important in order to increase productivity in maize (Zea mays L.). However, this control should be done rationally, prioritising environmentally safer methods such as biological control. This paper aims to report the occurrence of Dolichozele koebelei Viereck, 1911 (Hymenoptera: Braconidae) in Spodoptera frugiperda (J. E. Smith, 1797) (Lepidoptera: Noctuidae) larvae collected in maize subjected to different cropping systems. The experiment was conducted at the Centro Nacional de Pesquisa de Milho e Sorgo (CNPMS) in Sete Lagoas, Minas Gerais State, Brazil, using organic and conventional production. Ten plants were sampled from each of the 24 plots and for each production system, three times a week during the entire cycle of maize (variety BR 106). In the laboratory, larvae were distributed in individual rearing containers with artificial diet until the end of the biological cycle. An increased number of S. frugiperda larvae was observed in organic single crop maize; hence a higher percentage of S. frugiperda larvae parasitised by Hymenoptera and Diptera also occurred in the maize under this production system. Dolichozele koebelei had not yet been described in association with larvae of S. frugiperda. The percentage of parasitism of S. frugiperda larvae was high in both experiments, indicating the importance of natural control agents in reducing the population density of S. frugiperda, and especially the importance of an appropriate crop management. PMID:25627388

  19. Fitness costs of Cry1F resistance in two populations of fall armyworm, Spodoptera frugiperda (J.E. Smith), collected from Puerto Rico and Florida.

    PubMed

    Dangal, Vikash; Huang, Fangneng

    2015-05-01

    The development of resistance in target pest populations is a threat to the sustainability of transgenic crops expressing Bacillus thuringiensis (Bt) proteins. Fall armyworm, Spodoptera frugiperda (J.E. Smith), is a major target pest of Bt maize in North and South America. This insect is the first target pest that has developed field resistance to Bt maize at multiple locations in these regions. The objective of this study was to assess the fitness costs associated with the Cry1F resistance in two populations of S. frugiperda collected from Puerto Rico (RR-PR) and Florida (RR-FL). In the study, fitness costs were evaluated by comparing survival, growth, and developmental time of seven populations of S. frugiperda on (1) non-Bt meridic diet and (2) non-Bt maize leaf tissue and non-Bt diet. The seven populations were RR-PR, RR-FL, a Bt-susceptible strain (Bt-SS), and four F1 populations developed from reciprocal crosses between Bt-SS and the two resistant populations. Biological parameters measured were neonate-to-adult survivorship, neonate-to-adult developmental time, 10day larval weight on non-Bt maize leaf tissue, pupal weight, and sex ratios. Results of the study show that the Cry1F resistance in both RR-PR and RR-FL was associated with considerable fitness costs, especially for the Florida population. Compared to the Bt-susceptible population, RR-PR showed an average of 61.1% reduction in larval weight, 20.4% less in neonate-to-adult survivorship, and 3.7days delay in neonate-to-adult developmental time. These fitness costs for RR-FL were 66.9%, 31.7% and 4.4days, respectively. The fitness costs of RR-PR and RR-FL appeared to be non-recessive. The results indicate that a diversified genetic basis may exist for the Cry1F resistance in S. frugiperda. PMID:25791021

  20. Transgenic Cotton Plants Expressing Cry1Ia12 Toxin Confer Resistance to Fall Armyworm (Spodoptera frugiperda) and Cotton Boll Weevil (Anthonomus grandis)

    PubMed Central

    de Oliveira, Raquel S.; Oliveira-Neto, Osmundo B.; Moura, Hudson F. N.; de Macedo, Leonardo L. P.; Arraes, Fabrício B. M.; Lucena, Wagner A.; Lourenço-Tessutti, Isabela T.; de Deus Barbosa, Aulus A.; da Silva, Maria C. M.; Grossi-de-Sa, Maria F.

    2016-01-01

    Gossypium hirsutum (commercial cooton) is one of the most economically important fibers sources and a commodity crop highly affected by insect pests and pathogens. Several transgenic approaches have been developed to improve cotton resistance to insect pests, through the transgenic expression of different factors, including Cry toxins, proteinase inhibitors, and toxic peptides, among others. In the present study, we developed transgenic cotton plants by fertilized floral buds injection (through the pollen-tube pathway technique) using an DNA expression cassette harboring the cry1Ia12 gene, driven by CaMV35S promoter. The T0 transgenic cotton plants were initially selected with kanamycin and posteriorly characterized by PCR and Southern blot experiments to confirm the genetic transformation. Western blot and ELISA assays indicated the transgenic cotton plants with higher Cry1Ia12 protein expression levels to be further tested in the control of two major G. hirsutum insect pests. Bioassays with T1 plants revealed the Cry1Ia12 protein toxicity on Spodoptera frugiperda larvae, as evidenced by mortality up to 40% and a significant delay in the development of the target insects compared to untransformed controls (up to 30-fold). Also, an important reduction of Anthonomus grandis emerging adults (up to 60%) was observed when the insect larvae were fed on T1 floral buds. All the larvae and adult insect survivors on the transgenic lines were weaker and significantly smaller compared to the non-transformed plants. Therefore, this study provides GM cotton plant with simultaneous resistance against the Lepidopteran (S. frugiperda), and the Coleopteran (A. grandis) insect orders, and all data suggested that the Cry1Ia12 toxin could effectively enhance the cotton transgenic plants resistance to both insect pests. PMID:26925081

  1. ROLE OF SERUM AND ION CHANNEL BLOCK ON GROWTH AND HORMONALLY-INDUCED DIFFERENTIATION OF Spodoptera frugiperda (Sf21) INSECT CELLS.

    PubMed

    Jenson, Lacey J; Bloomquist, Jeffrey R

    2015-11-01

    A neuronal morphological phenotype can be induced in cultured Spodoptera frugiperda insect cells (Sf21) by supplementing serum-containing media with 20-hydroxyecdysone (20-HE) and/or insulin. In this study, the primary objectives were to determine any role of ion channels in mediating the morphological change in cells treated with 20-HE and insulin, and whether serum was required to observe this effect. Results showed serum-free media also induced growth of processes in Sf21 cells, but at a lower percentage than that found previously in cells bathed in serum-containing media. Veratridine, a sodium channel activator, increased cell survival when applied in combination with 20-HE to Sf21 cells, and the effect was blocked by tetrodotoxin (1 μM) a known sodium channel blocker. Cobalt, a calcium channel blocker, showed significant inhibition of cell process growth when applied in combination with both 20-HE and 20-HE plus veratridine. Cobalt also showed significant inhibition of cell process growth when applied in combination with insulin. Thus, some type of sodium channel, as well as a mechanism for transmembrane calcium ion movement, are apparently expressed in Sf21 cells and are involved in the differentiation process. These cell lines may be used in a wide variety of endeavors, including the screening of insecticides, as well as foster basic studies of neurodevelopment and ecdysone action. PMID:26183110

  2. Codon optimization of the rabbit hemorrhagic disease virus (RHDV) capsid gene leads to increased gene expression in Spodoptera frugiperda 9 (Sf9) cells

    PubMed Central

    Gao, Jingpeng; Meng, Chunchun; Chen, Zongyan; Li, Chuanfeng

    2013-01-01

    Rabbit hemorrhagic disease (RHD) is contagious and highly lethal. Commercial vaccines against RHD are produced from the livers of experimentally infected rabbits. Although several groups have reported that recombinant subunit vaccines against rabbit hemorrhagic disease virus (RHDV) are promising, application of the vaccines has been restricted due to high production costs or low yield. In the present study, we performed codon optimization of the capsid gene to increase the number of preference codons and eliminate rare codons in Spodoptera frugiperda 9 (Sf9) cells. The capsid gene was then subcloned into the pFastBac plasmid, and the recombinant baculoviruses were identified with a plaque assay. As expected, expression of the optimized capsid protein was markedly increased in the Sf9 cells, and the recombinant capsid proteins self-assembled into virus-like particles (VLPs) that were released into the cell supernatant. Rabbits inoculated with the supernatant and the purified VLPs were protected against RHDV challenge. A rapid, specific antibody response against RHDV was detected by an ELISA in all of the experimental groups. In conclusion, this strategy of producing a recombinant subunit vaccine antigen can be used to develop a low-cost, insect cell-derived recombinant subunit vaccine against RHDV. PMID:23820215

  3. Rhabdovirus-like endogenous viral elements in the genome of Spodoptera frugiperda insect cells are actively transcribed: Implications for adventitious virus detection.

    PubMed

    Geisler, Christoph; Jarvis, Donald L

    2016-07-01

    Spodoptera frugiperda (Sf) cell lines are used to produce several biologicals for human and veterinary use. Recently, it was discovered that all tested Sf cell lines are persistently infected with Sf-rhabdovirus, a novel rhabdovirus. As part of an effort to search for other adventitious viruses, we searched the Sf cell genome and transcriptome for sequences related to Sf-rhabdovirus. To our surprise, we found intact Sf-rhabdovirus N- and P-like ORFs, and partial Sf-rhabdovirus G- and L-like ORFs. The transcribed and genomic sequences matched, indicating the transcripts were derived from the genomic sequences. These appear to be endogenous viral elements (EVEs), which result from the integration of partial viral genetic material into the host cell genome. It is theoretically impossible for the Sf-rhabdovirus-like EVEs to produce infectious virus particles as 1) they are disseminated across 4 genomic loci, 2) the G and L ORFs are incomplete, and 3) the M ORF is missing. Our finding of transcribed virus-like sequences in Sf cells underscores that MPS-based searches for adventitious viruses in cell substrates used to manufacture biologics should take into account both genomic and transcribed sequences to facilitate the identification of transcribed EVE's, and to avoid false positive detection of replication-competent adventitious viruses. PMID:27236849

  4. Characterization of an Sf-rhabdovirus-negative Spodoptera frugiperda cell line as an alternative host for recombinant protein production in the baculovirus-insect cell system.

    PubMed

    Maghodia, Ajay B; Geisler, Christoph; Jarvis, Donald L

    2016-06-01

    Cell lines derived from the fall armyworm, Spodoptera frugiperda (Sf), are widely used as hosts for recombinant protein production in the baculovirus-insect cell system (BICS). However, it was recently discovered that these cell lines are contaminated with a virus, now known as Sf-rhabdovirus [1]. The detection of this adventitious agent raised a potential safety issue that could adversely impact the BICS as a commercial recombinant protein production platform. Thus, we examined the properties of Sf-RVN, an Sf-rhabdovirus-negative Sf cell line, as a potential alternative host. Nested RT-PCR assays showed Sf-RVN cells had no detectable Sf-rhabdovirus over the course of 60 passages in continuous culture. The general properties of Sf-RVN cells, including their average growth rates, diameters, morphologies, and viabilities after baculovirus infection, were virtually identical to those of Sf9 cells. Baculovirus-infected Sf-RVN and Sf9 cells produced equivalent levels of three recombinant proteins, including an intracellular prokaryotic protein and two secreted eukaryotic glycoproteins, and provided similar N-glycosylation patterns. In fact, except for the absence of Sf-rhabdovirus, the only difference between Sf-RVN and Sf9 cells was SF-RVN produced higher levels of infectious baculovirus progeny. These results show Sf-RVN cells can be used as improved, alternative hosts to circumvent the potential safety hazard associated with the use of Sf-rhabdovirus-contaminated Sf cells for recombinant protein manufacturing with the BICS. PMID:26923062

  5. Heritability of Wing Size and Shape of the Rice and Corn Strains of Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae).

    PubMed

    Cañas-Hoyos, N; Márquez, E J; Saldamando-Benjumea, C I

    2016-08-01

    Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) represents a pest of economic importance in all Western Hemisphere. This polyphagous species has diverged into two populations that have been mainly recognized with various mitochondrial and nuclear molecular markers and named "the rice" and "the corn" strains. In Colombia, both strains have evolved prezygotic and postzygotic isolation. They differ in tolerance to Bacillus thuringiensis (Cry1Ac and Cry1Ab endotoxins) and the insecticides lambda-cyhalothrin and methomyl. In 2014, a wing morphometric analysis made in 159 individuals from a colony showed that both strains significantly differ in wing shape. The species also exhibits sexual dimorphism in the rice strain as in females wing size is larger than in males. Here, we continued this work with another wing morphometric approach in laboratory-reared strains to calculate wing size and shape heritabilities using a full-sib design and in wild populations to determine if this method distinguishes these strains. Our results show that male heritabilities of both traits were higher than female ones. Wild populations were significantly different in wing shape and size. These results suggest that wing morphometrics can be used as an alternative method to molecular markers to differentiate adults from laboratory-reared populations and wild populations of this pest, particularly in males of this species. Finally, Q ST values obtained for wing size and shape further demonstrated that both strains are genetically differentiated in nature. PMID:27044394

  6. Genomic Sequence of Spodoptera frugiperda Ascovirus 1a, an Enveloped, Double-Stranded DNA Insect Virus That Manipulates Apoptosis for Viral Reproduction▿

    PubMed Central

    Bideshi, Dennis K.; Demattei, Marie-Véronique; Rouleux-Bonnin, Florence; Stasiak, Karine; Tan, Yeping; Bigot, Sylvie; Bigot, Yves; Federici, Brian A.

    2006-01-01

    Ascoviruses (family Ascoviridae) are double-stranded DNA viruses with circular genomes that attack lepidopterans, where they produce large, enveloped virions, 150 by 400 nm, and cause a chronic, fatal disease with a cytopathology resembling that of apoptosis. After infection, host cell DNA is degraded, the nucleus fragments, and the cell then cleaves into large virion-containing vesicles. These vesicles and virions circulate in the hemolymph, where they are acquired by parasitic wasps during oviposition and subsequently transmitted to new hosts. To develop a better understanding of ascovirus biology, we sequenced the genome of the type species Spodoptera frugiperda ascovirus 1a (SfAV-1a). The genome consisted of 156,922 bp, with a G+C ratio of 49.2%, and contained 123 putative open reading frames coding for a variety of enzymes and virion structural proteins, of which tentative functions were assigned to 44. Among the most interesting enzymes, due to their potential role in apoptosis and viral vesicle formation, were a caspase, a cathepsin B, several kinases, E3 ubiquitin ligases, and especially several enzymes involved in lipid metabolism, including a fatty acid elongase, a sphingomyelinase, a phosphate acyltransferase, and a patatin-like phospholipase. Comparison of SfAV-1a proteins with those of other viruses showed that 10% were orthologs of Chilo iridescent virus proteins, the highest correspondence with any virus, providing further evidence that ascoviruses evolved from a lepidopteran iridovirus. The SfAV-1a genome sequence will facilitate the determination of how ascoviruses manipulate apoptosis to generate the novel virion-containing vesicles characteristic of these viruses and enable study of their origin and evolution. PMID:16987980

  7. Strong oviposition preference for Bt over non-Bt maize in Spodoptera frugiperda and its implications for the evolution of resistance

    PubMed Central

    2014-01-01

    Background Transgenic crops expressing Bt toxins have substantial benefits for growers in terms of reduced synthetic insecticide inputs, area-wide pest management and yield. This valuable technology depends upon delaying the evolution of resistance. The ‘high dose/refuge strategy’, in which a refuge of non-Bt plants is planted in close proximity to the Bt crop, is the foundation of most existing resistance management. Most theoretical analyses of the high dose/refuge strategy assume random oviposition across refugia and Bt crops. Results In this study we examined oviposition and survival of Spodoptera frugiperda across conventional and Bt maize and explored the impact of oviposition behavior on the evolution of resistance in simulation models. Over six growing seasons oviposition rates per plant were higher in Bt crops than in refugia. The Cry1F Bt maize variety retained largely undamaged leaves, and oviposition preference was correlated with the level of feeding damage in the refuge. In simulation models, damage-avoiding oviposition accelerated the evolution of resistance and either led to requirements for larger refugia or undermined resistance management altogether. Since larval densities affected oviposition preferences, pest population dynamics affected resistance evolution: larger refugia were weakly beneficial for resistance management if they increased pest population sizes and the concomitant degree of leaf damage. Conclusions Damaged host plants have reduced attractiveness to many insect pests, and crops expressing Bt toxins are generally less damaged than conventional counterparts. Resistance management strategies should take account of this behavior, as it has the potential to undermine the effectiveness of existing practice, especially in the tropics where many pests are polyvoltinous. Efforts to bring down total pest population sizes and/or increase the attractiveness of damaged conventional plants will have substantial benefits for slowing the

  8. Synergism and Antagonism between Bacillus thuringiensis Vip3A and Cry1 Proteins in Heliothis virescens, Diatraea saccharalis and Spodoptera frugiperda

    PubMed Central

    Lemes, Ana Rita Nunes; Davolos, Camila Chiaradia; Legori, Paula Cristina Brunini Crialesi; Fernandes, Odair Aparecido; Ferré, Juan; Lemos, Manoel Victor Franco; Desiderio, Janete Apparecida

    2014-01-01

    Second generation Bt crops (insect resistant crops carrying Bacillus thuringiensis genes) combine more than one gene that codes for insecticidal proteins in the same plant to provide better control of agricultural pests. Some of the new combinations involve co-expression of cry and vip genes. Because Cry and Vip proteins have different midgut targets and possibly different mechanisms of toxicity, it is important to evaluate possible synergistic or antagonistic interactions between these two classes of toxins. Three members of the Cry1 class of proteins and three from the Vip3A class were tested against Heliothis virescens for possible interactions. At the level of LC50, Cry1Ac was the most active protein, whereas the rest of proteins tested were similarly active. However, at the level of LC90, Cry1Aa and Cry1Ca were the least active proteins, and Cry1Ac and Vip3A proteins were not significantly different. Under the experimental conditions used in this study, we found an antagonistic effect of Cry1Ca with the three Vip3A proteins. The interaction between Cry1Ca and Vip3Aa was also tested on two other species of Lepidoptera. Whereas antagonism was observed in Spodoptera frugiperda, synergism was found in Diatraea saccharalis. In all cases, the interaction between Vip3A and Cry1 proteins was more evident at the LC90 level than at the LC50 level. The fact that the same combination of proteins may result in a synergistic or an antagonistic interaction may be an indication that there are different types of interactions within the host, depending on the insect species tested. PMID:25275646

  9. Susceptibility of Anthonomus grandis (cotton boll weevil) and Spodoptera frugiperda (fall armyworm) to a cry1ia-type toxin from a Brazilian Bacillus thuringiensis strain.

    PubMed

    Grossi-de-Sa, Maria Fatima; Quezado de Magalhaes, Mariana; Silva, Marilia Santos; Silva, Shirley Margareth Buffon; Dias, Simoni Campos; Nakasu, Erich Yukio Tempel; Brunetta, Patricia Sanglard Felipe; Oliveira, Gustavo Ramos; Neto, Osmundo Brilhante de Oliveira; Sampaio de Oliveira, Raquel; Soares, Luis Henrique Barros; Ayub, Marco Antonio Zachia; Siqueira, Herbert Alvaro Abreu; Figueira, Edson L Z

    2007-09-30

    Different isolates of the soil bacterium Bacillus thuringiensis produce multiple crystal (Cry) proteins toxic to a variety of insects, nematodes and protozoans. These insecticidal Cry toxins are known to be active against specific insect orders, being harmless to mammals, birds, amphibians, and reptiles. Due to these characteristics, genes encoding several Cry toxins have been engineered in order to be expressed by a variety of crop plants to control insectpests. The cotton boll weevil, Anthonomus grandis, and the fall armyworm, Spodoptera frugiperda, are the major economically devastating pests of cotton crop in Brazil, causing severe losses, mainly due to their endophytic habit, which results in damages to the cotton boll and floral bud structures. A cry1Ia-type gene, designated cry1Ia12, was isolated and cloned from the Bt S811 strain. Nucleotide sequencing of the cry1Ia12 gene revealed an open reading frame of 2160 bp, encoding a protein of 719 amino acid residues in length, with a predicted molecular mass of 81 kDa. The amino acid sequence of Cry1Ia12 is 99% identical to the known Cry1Ia proteins and differs from them only in one or two amino acid residues positioned along the three domains involved in the insecticidal activity of the toxin. The recombinant Cry1Ia12 protein, corresponding to the cry1Ia12 gene expressed in Escherichia coli cells, showed moderate toxicity towards first instar larvae of both cotton boll weevil and fall armyworm. The highest concentration of the recombinant Cry1Ia12 tested to achieve the maximum toxicities against cotton boll weevil larvae and fall armyworm larvae were 230 microg/mL and 5 microg/mL, respectively. The herein demonstrated insecticidal activity of the recombinant Cry1Ia12 toxin against cotton boll weevil and fall armyworm larvae opens promising perspectives for the genetic engineering of cotton crop resistant to both these devastating pests in Brazil. PMID:17927912

  10. Evaluation of Extracellular Subviral Particles of Dengue Virus Type 2 and Japanese Encephalitis Virus Produced by Spodoptera frugiperda Cells for Use as Vaccine and Diagnostic Antigens ▿

    PubMed Central

    Kuwahara, Miwa; Konishi, Eiji

    2010-01-01

    New or improved vaccines against dengue virus types 1 to 4 (DENV1 to DENV4) and Japanese encephalitis virus (JEV), the causative agents of dengue fever and Japanese encephalitis (JE), respectively, are urgently required. The use of noninfectious subviral extracellular particles (EPs) is an inexpensive and safe strategy for the production of protein-based flavivirus vaccines. Although coexpression of premembrane (prM) and envelope (E) proteins has been demonstrated to produce EPs in mammalian cells, low yields have hindered their commercial application. Therefore, we used an insect cell expression system with Spodoptera frugiperda-derived Sf9 cells to investigate high-level production of DENV2 and JEV EPs. Sf9 cells transfected with the prM and E genes of DENV2 or JEV secreted corresponding viral antigens in a particulate form that were biochemically and biophysically equivalent to the authentic antigens obtained from infected C6/36 mosquito cells. Additionally, equivalent neutralizing antibody titers were induced in mice immunized either with EPs produced by transfected Sf9 cells or with EPs produced by transfected mammalian cells, in the context of coimmunization with a DNA vaccine that expresses EPs. Furthermore, the results of an enzyme-linked immunosorbent assay (ELISA) using an EP antigen derived from Sf9 cells correlated significantly with the results obtained by a neutralization test and an ELISA using an EP antigen derived from mammalian cells. Finally, Sf9 cells could produce 10- to 100-fold larger amounts of E antigen than mammalian cells. These results indicate the potential of Sf9 cells for high-level production of flavivirus protein vaccines and diagnostic antigens. PMID:20668137

  11. Protective efficacy of Anopheles minimus CYP6P7 and CYP6AA3 against cytotoxicity of pyrethroid insecticides in Spodoptera frugiperda (Sf9) insect cells.

    PubMed

    Duangkaew, P; Kaewpa, D; Rongnoparut, P

    2011-08-01

    Cytochrome P450 monooxygenases (P450s) are enzymes known to metabolize a wide variety of compounds including insecticides. Their overexpression leading to enhanced insecticide detoxification could result in insecticide resistance in insects. The increased mRNA expression of two P450 genes, CYP6P7 and CYP6AA3, has been previously observed in laboratory-selected deltamethrin-resistant Anopheles minimus, a major malaria vector in Southeast Asia, suggesting their role in detoxification of pyrethroids. In this study CYP6P7 and CYP6AA3 were expressed in insect Spodoptera frugiperda (Sf9) cells via baculovirusdirected expression system. Insecticide detoxification capabilities of Sf9 cells with and without expression of CYP6P7 or CYP6AA3 were evaluated using 3-(4,5-dimethyl-thiazol-2- yl)-2,5-diphenyltetrazolium bromide (MTT) assays. The results revealed that CYP6P7- or CYP6AA3-expressing cells showed significantly higher cytoprotective capability than parental Sf9 cells against cytotoxicity of pyrethroids including permethrin, cypermethrin and deltamethrin. Such cytoprotective effect was not observed for bioallethrin (pyrethroid), chlorpyrifos (organophosphate) and propoxur (carbamate). Moreover, expression of CYP6AA3, but not CYP6P7, could protect cells against λ-cyhalothrin cytotoxicity. In MTT assays upon co-incubation with piperonyl butoxide (P450 inhibitor), cytoprotective ability of CYP6P7 and CYP6AA3 against deltamethrin was diminished, implying that pyrethroid detoxification was due to activities of P450 enzymes. Insecticide detoxification capabilities of CYP6P7 and CYP6AA3 observed from MTT assays were correlated to their pyrethroid metabolizing activities observed from in vitro reconstitution enzymatic assays. Thus MTT assays using cells expressing P450 enzymes of interest could be primarily used to determine detoxification activities of enzymes against cytotoxic insecticides. PMID:22041748

  12. Azadirachtin-induced apoptosis involves lysosomal membrane permeabilization and cathepsin L release in Spodoptera frugiperda Sf9 cells.

    PubMed

    Wang, Zheng; Cheng, Xingan; Meng, Qianqian; Wang, Peidan; Shu, Benshui; Hu, Qiongbo; Hu, Meiying; Zhong, Guohua

    2015-07-01

    Azadirachtin as a kind of botanical insecticide has been widely used in pest control. We previously reported that azadirachtin could induce apoptosis of Spodoptera litura cultured cell line Sl-1, which involves in the up-regulation of P53 protein. However, the detailed mechanism of azadirachtin-induced apoptosis is not clearly understood in insect cultured cells. The aim of the present study was to address the involvement of lysosome and lysosomal protease in azadirachtin-induced apoptosis in Sf9 cells. The result confirmed that azadirachtin indeed inhibited proliferation and induced apoptosis. The lysosomes were divided into different types as time-dependent manner, which suggested that changes of lysosomes were necessarily physiological processes in azadirachtin-induced apoptosis in Sf9 cells. Interestingly, we noticed that azadirachtin could trigger lysosomal membrane permeabilization and cathepsin L releasing to cytosol. Z-FF-FMK (a cathepsin L inhibitor), but not CA-074me (a cathepsin B inhibitor), could effectively hinder the apoptosis induced by azadirachtin in Sf9 cells. Meanwhile, the activity of caspase-3 could also be inactivated by the inhibition of cathepsin L enzymatic activity induced by Z-FF-FMK. Taken together, our findings suggest that azadirachtin could induce apoptosis in Sf9 cells in a lysosomal pathway, and cathepsin L plays a pro-apoptosis role in this process through releasing to cytosol and activating caspase-3. PMID:25849458

  13. SEARCHING FOR EXOTIC SPODOPTERA SPECIES

    Technology Transfer Automated Retrieval System (TEKTRAN)

    We used sex pheromone-baited traps to collect native and exotic Spodoptera spp. moths at an orchid nursery in Lake County, FL. Lures for S. eridania, exempta, exigua, frugiperda, littoralis, litura, praefica, and Pseudaletia unipuncta were placed in bucket traps that surrounded the greenhouses of t...

  14. Shared midgut binding sites for Cry1A.105, Cry1Aa, Cry1Ab, Cry1Ac and Cry1Fa proteins from Bacillus thuringiensis in two important corn pests, Ostrinia nubilalis and Spodoptera frugiperda.

    PubMed

    Hernández-Rodríguez, Carmen Sara; Hernández-Martínez, Patricia; Van Rie, Jeroen; Escriche, Baltasar; Ferré, Juan

    2013-01-01

    First generation of insect-protected transgenic corn (Bt-corn) was based on the expression of Cry1Ab or Cry1Fa proteins. Currently, the trend is the combination of two or more genes expressing proteins that bind to different targets. In addition to broadening the spectrum of action, this strategy helps to delay the evolution of resistance in exposed insect populations. One of such examples is the combination of Cry1A.105 with Cry1Fa and Cry2Ab to control O. nubilalis and S. frugiperda. Cry1A.105 is a chimeric protein with domains I and II and the C-terminal half of the protein from Cry1Ac, and domain III almost identical to Cry1Fa. The aim of the present study was to determine whether the chimeric Cry1A.105 has shared binding sites either with Cry1A proteins, with Cry1Fa, or with both, in O. nubilalis and in S. frugiperda. Brush-border membrane vesicles (BBMV) from last instar larval midguts were used in competition binding assays with (125)I-labeled Cry1A.105, Cry1Ab, and Cry1Fa, and unlabeled Cry1A.105, Cry1Aa, Cry1Ab, Cry1Ac, Cry1Fa, Cry2Ab and Cry2Ae. The results showed that Cry1A.105, Cry1Ab, Cry1Ac and Cry1Fa competed with high affinity for the same binding sites in both insect species. However, Cry2Ab and Cry2Ae did not compete for the binding sites of Cry1 proteins. Therefore, according to our results, the development of cross-resistance among Cry1Ab/Ac, Cry1A.105, and Cry1Fa proteins is possible in these two insect species if the alteration of shared binding sites occurs. Conversely, cross-resistance between these proteins and Cry2A proteins is very unlikely in such case. PMID:23861865

  15. The synthesis and characterization of cellular membrane affinity chromatography columns for the study of human multidrug resistant proteins MRP1, MRP2 and human breast cancer resistant protein BCRP using membranes obtained from Spodoptera frugiperda (Sf9) insect cells

    PubMed Central

    Bhatia, Prateek A.; Moaddel, Ruin; Wainer, Irving W.

    2010-01-01

    CMAC (cellular membrane affinity chromatography columns) have been developed for the study of the human multidrug transporters MRP1, MRP2 and the breast cancer resistance protein (BCRP). The columns were constructed using the immobilized artificial membrane (IAM) stationary phase and cellular membrane fragments obtained from Spodopetra frugiperda (Sf9) cells that had been stably transfected with human Mrp1, Mrp2 or Bcrp c-DNA, using a baculovirus expression system. The resulting CMAC(Sf9MRP1), CMAC(Sf9MRP2) and CMAC(Sf9BCRP) columns and a control column produced using membrane fragments from non-transfected Sf9 cells, CMAC(Sf9), were characterized using frontal affinity chromatography using [3H]-etoposide as the marker ligand and etoposide, benzbromarone and MK571 as the displacers on the CMAC(Sf9MRP1) column, etoposide and furosemide on the CMAC(Sf9MRP2) column and etoposide and fumitremorgin C on the CMAC(Sf9BCPR) column The binding affinities (Ki values) obtained from the chromatographic studies were consistent with the data obtained using non-chromatographic techniques and the results indicate that the immobilized MRP1, MRP2 and BCRP transporters retained their ability to selectively bind known ligands. (S)-verapamil displaced [3H]-etoposide on the CMAC(Sf9MRP1) column to a greater extent than (R)-verapamil and the relative IC50 values of the enantiomers were calculated using the changes in the retention times of the marker. The observed enantioselectivity and calculated IC50 values were consistent with previously reported data. The results indicated that the CMAC(Sf9MRP1), CMAC(Sf9MRP2) and CMAC(Sf9BCRP) columns can be used for the study of binding to the MRP1, MRP2 and BCRP transporters and that membranes from the Sf9 cell line can be used to prepare CMAC columns. This is the first example of the use of membranes from a non-mammalian cell line in an affinity chromatographic system. PMID:20441926

  16. Telenomus remus Nixon egg parasitization of three species of Spodoptera under different temperatures.

    PubMed

    Pomari, A F; Bueno, A F; Bueno, R C O F; Menezes, A O

    2013-08-01

    Telenomus remus Nixon is a promising biocontrol agent as an egg parasitoid of Spodoptera spp., but the lack of information on the host-parasitoid interactions in this system precludes its applied use in agriculture. Therefore, we studied the parasitism capacity of T. remus on eggs of Spodoptera cosmioides (Walker), Spodoptera eridania (Cramer), and Spodoptera frugiperda (Smith) in a range of temperatures (19, 22, 25, 28, 31, and 34 ± 1°C) under controlled conditions (70 ± 10% RH and 12 h photophase). Egg masses of Spodoptera spp. were offered to a single-mated T. remus female on a daily basis. More than 80% lifetime parasitism on eggs of S. cosmioides, S. frugiperda, and S. eridania was reached from 1 to 5, 1 to 7, and 1 to 9 days, respectively, at temperatures from 19 to 34°C. More than 80% parasitization was obtained at extreme temperatures for all hosts studied. Lifetime parasitization of S. frugiperda, S. cosmioides, and S. eridania was affected by temperature, with the lowest values for S. frugiperda (34°C) and S. cosmioides (19 and 34°C). Parasitization of S. eridania eggs was reduced around 18% at 28 and 31°C, but dropped more severely at 34°C. Parasitoid longevity was reduced as temperature increased. Thus, our data indicated that T. remus might be suitable as a biocontrol agent against S. eridania, S. cosmioides, and S. frugiperda in geographical areas that fit the temperature range studied here, even though T. remus parasitism was reduced at 34°C. PMID:23949860

  17. SPODOBASE : an EST database for the lepidopteran crop pest Spodoptera

    PubMed Central

    Nègre, Vincent; Hôtelier, Thierry; Volkoff, Anne-Nathalie; Gimenez, Sylvie; Cousserans, François; Mita, Kazuei; Sabau, Xavier; Rocher, Janick; López-Ferber, Miguel; d'Alençon, Emmanuelle; Audant, Pascaline; Sabourault, Cécile; Bidegainberry, Vincent; Hilliou, Frédérique; Fournier, Philippe

    2006-01-01

    Background The Lepidoptera Spodoptera frugiperda is a pest which causes widespread economic damage on a variety of crop plants. It is also well known through its famous Sf9 cell line which is used for numerous heterologous protein productions. Species of the Spodoptera genus are used as model for pesticide resistance and to study virus host interactions. A genomic approach is now a critical step for further new developments in biology and pathology of these insects, and the results of ESTs sequencing efforts need to be structured into databases providing an integrated set of tools and informations. Description The ESTs from five independent cDNA libraries, prepared from three different S. frugiperda tissues (hemocytes, midgut and fat body) and from the Sf9 cell line, are deposited in the database. These tissues were chosen because of their importance in biological processes such as immune response, development and plant/insect interaction. So far, the SPODOBASE contains 29,325 ESTs, which are cleaned and clustered into non-redundant sets (2294 clusters and 6103 singletons). The SPODOBASE is constructed in such a way that other ESTs from S. frugiperda or other species may be added. User can retrieve information using text searches, pre-formatted queries, query assistant or blast searches. Annotation is provided against NCBI, UNIPROT or Bombyx mori ESTs databases, and with GO-Slim vocabulary. Conclusion The SPODOBASE database provides integrated access to expressed sequence tags (EST) from the lepidopteran insect Spodoptera frugiperda. It is a publicly available structured database with insect pest sequences which will allow identification of a number of genes and comprehensive cloning of gene families of interest for scientific community. SPODOBASE is available from URL: PMID:16796757

  18. Differential Resistance of Switchgrass Cultivars to Fall Armyworms (Spodoptera frugiperda)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Switchgrass has great potential as a sustainable bioenergy crop. However, prior reports suggest insect damage can interfere with switchgrass establishment and yield. The fall armyworm is a pest of many different grass crops (including maize and rice) and is thus potentially an important pest of sw...

  19. Diet dependent metabolic responses in three generalist insect herbivores Spodoptera spp.

    PubMed

    Roy, A; Walker, W B; Vogel, H; Chattington, S; Larsson, M C; Anderson, P; Heckel, D G; Schlyter, F

    2016-04-01

    Adaption to dietary changes is critical in the evolution of host plant ranges in polyphagous insects. We compared three taxa of lepidopteran herbivores from the predominantly generalist genus Spodoptera showing different degrees of polyphagy: Spodoptera littoralis, with a broad host range including both mono- and dicotyledonous plants, and two Spodoptera frugiperda strains [Corn (i.e. maize) (C) and Rice (R)] adapted primarily to different grass species. When feeding on maize we show a lower performance in the broad generalist taxon compared to the grass adapted taxa. Among these taxa, the maize adapted S. frugiperda C-strain generally performed better than the R-strain on maize leaves. On artificial pinto diet, all taxa performed well. Our RNA-Seq analysis of midgut transcriptomes from 3rd instar larvae feeding on maize showed broader transcriptional readjustments in the generalist S. littoralis compared to grass adapted S. frugiperda strains. Substantial alteration in the expression levels of midgut physiological function related transcripts, such as digestive and detoxifying enzymes, transporters, immunity, and peritrophic membrane associated transcripts, existed in all taxa. We found high background expression of UDP-glucosyl transferases, which are known to neutralize maize leaf toxins, in the maize adapted S. frugiperda C-strain, contributing to its fitness on maize compared to the R-strain. Our findings provide evidence for divergent diet specific response of digestive physiology within these Spodoptera taxa. Unexpectedly, the C- and R-strains of S. frugiperda fed on the same diet showed large differences in expression patterns between these two closely related taxa. PMID:26908076

  20. A noda-like virus isolated from the sweetpotato pest spodoptera eridania (Cramer) (Lep.; noctuidae)

    PubMed

    Zeddam; Rodriguez; Ravallec; Lagnaoui

    1999-11-01

    A small isometric virus has been isolated from larvae of the sweetpotato pest Spodoptera eridania (Cramer) collected near Pariacoto, Ancash province, Peru. It is designated the Pariacoto virus (PaV). In addition to its high pathogenicity on its natural host Spodoptera eridania, PaV was found to replicate in Spodoptera ochrea (Hampson) larvae but not in Spodoptera frugiperda (Smith) larvae. The size of the viral particle was estimated to be about 30 nm in diameter. Polyacrylamide gel electrophoresis showed a protein of approximately 40.5 kDa. After agarose gel electrophoresis, the viral genome appeared to be bipartite RNA. Gel immunodiffusion tests showed no serological relationship between PaV and Nodamura virus, the type species for insect nodaviruses. Electron microscopy confirmed that viral replication occurs in the cytoplasm. These properties are similar to those of other members of family Nodaviridae, to which the virus is currently assigned. Copyright 1999 Academic Press. PMID:10534414

  1. Bacillus thuringiensis Cry1Ia10 and Vip3Aa protein interactions and their toxicity in Spodoptera spp. (Lepidoptera).

    PubMed

    Bergamasco, V B; Mendes, D R P; Fernandes, O A; Desidério, J A; Lemos, M V F

    2013-02-01

    The polyphagous pests belonging to the genus Spodoptera are considered to be among the most important causes of damage and are widely distributed throughout the Americas'. Due to the extensive use of genetically modified plants containing Bacillus thuringiensis genes that code for insecticidal proteins, resistant insects may arise. To prevent the development of resistance, pyramided plants, which express multiple insecticidal proteins that act through distinct mode of actions, can be used. This study analyzed the mechanisms of action for the proteins Cry1Ia10 and Vip3Aa on neonatal Spodoptera frugiperda, Spodoptera albula, Spodoptera eridania and Spodoptera cosmioides larvae. The interactions of these toxins with receptors on the intestinal epithelial membrane were also analyzed by binding biotinylated toxins to brush border membrane vesicles (BBMVs) from the intestines of these insects. A putative receptor of approximately 65 kDa was found by ligand blotting in all of these species. In vitro competition assays using biotinylated proteins have indicated that Vip3Aa and Cry1Ia10 do not compete for the same receptor for S. frugiperda, S. albula and S. cosmioides and that Vip3Aa was more efficient than Cry1Ia10 when tested individually, by bioassays. A synergistic effect of the toxins in S. frugiperda, S. albula and S. cosmioides was observed when they were combined. However, in S. eridania, Cry1Ia10 and Vip3Aa might compete for the same receptor and through bioassays Cry1Ia10 was more efficient than Vip3Aa and showed an antagonistic effect when the proteins were combined. These results suggest that using these genes to develop pyramided plants may not prove effective in preventing the development of resistance in S. eridiana. PMID:23220241

  2. [Approach to Spodoptera (Lepidoptera: Noctuidae) phylogeny based on the sequence of the cytocrhome oxydase I (COI) mitochondrial gene].

    PubMed

    Saldamando, Clara Inés; Marquez, Edna Judith

    2012-09-01

    The genus Spodoptera includes 30 species of moths considered important pests worldwide, with a great representation in the Western Hemisphere. In general, Noctuidae species have morphological similarities that have caused some difficulties for assertive species identification by conventional methods. The purpose of this work was to generate an approach to the genus phylogeny from several species of the genus Spodoptera and the species Bombyx mori as an out group, with the use of molecular tools. For this, a total of 102 S. frugiperda larvae were obtained at random in corn, cotton, rice, grass and sorghum, during late 2006 and early 2009, from Colombia. We took ADN samples from the larval posterior part and we analyzed a fragment of 451 base pairs of the mitochondrial gene cytochrome oxydase I (COI), to produce a maximum likelihood (ML) tree by using 62 sequences (29 Colombian haplotypes were used). Our results showed a great genetic differentiation (K2 distances) amongst S. frugiperda haplotypes from Colombia and the United States, condition supported by the estimators obtained for haplotype diversity and polymorphism. The obtained ML tree clustered most of the species with bootstrapping values from 73-99% in the interior branches; with low values also observed in some of the branches. In addition, this tree clustered two species of the Eastern hemisphere (S littoralis and S. litura) and eight species of the Western hemisphere (S. androgea, S. dolichos, S. eridania, S. exigua, S. frugiperda, S. latifascia, S. ornithogalli and S. pulchella). In Colombia, S. frugiperda, S. ornithogalli and S. albula represent a group of species referred as "the Spodoptera complex" of cotton crops, and our work demonstrated that sequencing a fragment of the COI gene, allows researchers to differentiate the first two species, and thus it can be used as an alternative method to taxonomic keys based on morphology. Finally, the ML tree did not cluster S. frugiperda with S. ornithogalli

  3. Cry1F resistance in fall armyworm Spodoptera frugiperda: single gene versus pyramided Bt maize

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Evolution of insect resistance to transgenic crops containing Bacillus thuringiensis (Bt) genes is a serious threat to the sustainability of this technology. However, field resistance related to the reduced efficacy of Bt maize has not been documented in any lepidopteran pest in the mainland U.S. af...

  4. Proteotoxic stress induced by Autographa californica nucleopolyhedrovirus infection of Spodoptera frugiperda Sf9 cells.

    PubMed

    Lyupina, Yulia V; Abaturova, Svetlana B; Erokhov, Pavel A; Orlova, Olga V; Beljelarskaya, Svetlana N; Mikhailov, Victor S

    2013-02-01

    Baculovirus AcMNPV causes proteotoxicity in Sf9 cells as revealed by accumulation of ubiquitinated proteins and aggresomes in the course of infection. Inhibition of proteasomes by lactacystin increased markedly the stock of ubiquitinated proteins indicating a primary role of proteasomes in detoxication. The proteasomes were present in Sf9 cells as 26S and 20S complexes whose protease activity did not change during infection. Proteasome inhibition caused a delay in the initiation of viral DNA replication suggesting an important role of proteasomes at early stages in infection. However, lactacystin did not affect ongoing replication indicating that active proteasomes are not required for genome amplification. At late stages in infection (24-48 hpi), aggresomes containing the ubiquitinated proteins and HSP/HSC70s showed gradual fusion with the vacuole-like structures identified as lysosomes by antibody to cathepsin D. This result suggests that lysosomes may assist in protection against proteotoxicity caused by baculoviruses absorbing the ubiquitinated proteins. PMID:23123012

  5. Molecular characterization of host strains of Spodoptera frugiperda(Lepidoptera:Noctuidae)in southern Brazil :

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fall armyworm infestations in most of North America north of Mexico arise from annual migrations of populations that overwinter in southern Texas and Florida. Mapping the pattern of migration and the relative contributions of the Texas and Florida populations would contribute both to our understandi...

  6. Migratory patterns of the fall armyworm (Spodoptera frugiperda) in the western hemisphere

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fall armyworm (FAW) is a serious pest of sweet corn in south Florida and a pest of other vegetable, row, and forage crops in the southeastern, mid-Atlantic, and central U.S. It is a migratory pest, moving north each season from overwintering areas in southern Texas and southern Florida. For the la...

  7. Proteotoxic stress induced by Autographa californica nucleopolyhedrovirus infection of Spodoptera frugiperda Sf9 cells

    SciTech Connect

    Lyupina, Yulia V.; Abaturova, Svetlana B.; Erokhov, Pavel A.; Orlova, Olga V.; Beljelarskaya, Svetlana N.; Mikhailov, Victor S.

    2013-02-05

    Baculovirus AcMNPV causes proteotoxicity in Sf9 cells as revealed by accumulation of ubiquitinated proteins and aggresomes in the course of infection. Inhibition of proteasomes by lactacystin increased markedly the stock of ubiquitinated proteins indicating a primary role of proteasomes in detoxication. The proteasomes were present in Sf9 cells as 26S and 20S complexes whose protease activity did not change during infection. Proteasome inhibition caused a delay in the initiation of viral DNA replication suggesting an important role of proteasomes at early stages in infection. However, lactacystin did not affect ongoing replication indicating that active proteasomes are not required for genome amplification. At late stages in infection (24-48 hpi), aggresomes containing the ubiquitinated proteins and HSP/HSC70s showed gradual fusion with the vacuole-like structures identified as lysosomes by antibody to cathepsin D. This result suggests that lysosomes may assist in protection against proteotoxicity caused by baculoviruses absorbing the ubiquitinated proteins.

  8. In vitro correction of disorders of lysosomal transport by microvesicles derived from baculovirus-infected Spodoptera cells.

    PubMed

    Thoene, Jess; Goss, Thomas; Witcher, Marc; Mullet, Jodi; N'Kuli, Francisca; Van Der Smissen, Patrick; Courtoy, Pierre; Hahn, Si Houn

    2013-05-01

    Infection of Spodoptera frugiperda (Sf9) cells by baculovirus (BV) is well established for transgene expression of soluble proteins, but few correctly folded transmembrane proteins have been so produced. We here report the use of the BV/Sf9 (BVES) method for the expression and transfer, via microvesicles, of the exclusive lysosomal exporters for cystine and sialic acid, human cystinosin and sialin. These proteins and their mRNA are released into the culture medium as very low-density microvesicles (~1.05 g/ml), which do not label for lysobisphosphatidic acid. The presence of the human transgene proteins in the vesicles was confirmed by western blotting and confirmed and quantified by mass spectrometry. Addition of vesicles to cultures of human fibroblast lines deficient in either cystinosin or sialin produced a progressive depletion of stored lysosomal cystine or sialic acid, respectively. The depletion effect was slow (T1/2 ~48 h), saturable (down to ~40% of initial after 4 days) and stable (>one week). Surprisingly, BV infection of Spodoptera appeared to induce expression and release into microvesicles of the insect orthologue of cystinosin, but not of sialin. We conclude that BVES is an effective method to express and transfer functional transmembrane proteins so as to study their properties in mammalian cells, and has a generic potential for transport protein replacement therapy. PMID:23465695

  9. Isolation of an Apoptosis Suppressor Gene of the Spodoptera littoralis Nucleopolyhedrovirus†

    PubMed Central

    Du, Quansheng; Lehavi, Dana; Faktor, Ouriel; Qi, Yipeng; Chejanovsky, Nor

    1999-01-01

    Spodoptera frugiperda SF9 cells infected with mutants of the Autographa californica nucleopolyhedrovirus (AcMNPV) which lack a functional p35 gene undergo apoptosis, aborting the viral infection. The Spodoptera littoralis nucleopolyhedrovirus (SlNPV) was able to suppress apoptosis triggered by vΔP35K/pol+, an AcMNPV p35 null mutant. To identify the putative apoptotic suppressor gene of SlNPV, overlapping cosmid clones representing the entire SlNPV genome were individually cotransfected along with genomic DNA of vΔP35K/pol+. Using this complementation assay, we isolated a SlNPV DNA fragment that was able to rescue the vΔP35K/pol+ infection in SF9 cells. By further subcloning and rescue, we identified a novel SlNPV gene, Slp49. The Slp49 sequence predicted a 49-kDa polypeptide with about 48.8% identity to the AcMNPV apoptotic suppressor P35. SLP49 displays a potential recognition site, TVTDG, for cleavage by death caspases. Recombinant AcMNPVs deficient in p35 bearing the Slp49 gene did not induce apoptosis and showed successful productive infections in SF9 cells, indicating that Slp49 is a functional homologue of p35. A 1.5-kbp Slp49-specific transcript was identified in SF9 cells infected with SlNPV or with vAc496, a vΔP35K/pol+-recombinant bearing Slp49. The discovery of Slp49 contributes to the identification of important functional motifs conserved in p35-like apoptotic suppressors and to the future isolation of p35-like genes from other baculoviruses. PMID:9882332

  10. Susceptibility and aversion of Spodoptera frugiperda to Cry1F Bt maize and considerations for insect resistance management

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bacillus thuringiensis (Bt) maize was developed primarily for North American pests such as European corn borer (Ostrinia nubilalis Hubner). However, most Bt maize products also are cultivated outside of North America, where the primary pests are different and often have lower susceptibility to Bt to...

  11. Enhanced Production of Chikungunya Virus-Like Particles Using a High-pH Adapted Spodoptera frugiperda Insect Cell Line

    PubMed Central

    Wagner, James M.; Pajerowski, J. David; Daniels, Christopher L.; McHugh, Patrick M.; Flynn, Jessica A.; Balliet, John W.; Casimiro, Danilo R.; Subramanian, Shyamsundar

    2014-01-01

    Chikungunya virus-like particles (VLPs) have potential to be used as a prophylactic vaccine based on testing in multiple animal models and are currently being evaluated for human use in a Phase I clinical trial. The current method for producing these enveloped alphavirus VLPs by transient gene expression in mammalian cells presents challenges for scalable and robust industrial manufacturing, so the insect cell baculovirus expression vector system was evaluated as an alternative expression technology. Subsequent to recombinant baculovirus infection of Sf21 cells in standard culture media (pH 6.2–6.4), properly processed Chikungunya structural proteins were detected and assembled capsids were observed. However, an increase in culture pH to 6.6–6.8 was necessary to produce detectable concentrations of assembled VLPs. Since this elevated production pH exceeds the optimum for growth medium stability and Sf21 culture, medium modifications were made and a novel insect cell variant (SfBasic) was derived by exposure of Sf21 to elevated culture pH for a prolonged period of time. The high-pH adapted SfBasic insect cell line described herein is capable of maintaining normal cell growth into the typical mammalian cell culture pH range of 7.0–7.2 and produces 11-fold higher Chikungunya VLP yields relative to the parental Sf21 cell line. After scale-up into stirred tank bioreactors, SfBasic derived VLPs were chromatographically purified and shown to be similar in size and structure to a VLP standard derived from transient gene expression in HEK293 cells. Total serum anti-Chikungunya IgG and neutralizing titers from guinea pigs vaccinated with SfBasic derived VLPs or HEK293 derived VLPs were not significantly different with respect to production method, suggesting that this adapted insect cell line and production process could be useful for manufacturing Chikungunya VLPs for use as a vaccine. The adaptation of Sf21 to produce high levels of recombinant protein and VLPs in an elevated pH range may also have applications for other pH-sensitive protein or VLP targets. PMID:24713807

  12. Using satellite multispectral imagery for damage mapping armyworm (Spodoptera frugiperda) in maize damage at a regional scale

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Armyworm, as a destructive insect for maize, causes wide range of damage in both China and U.S. in recent years. To obtain the spatial distribution of damage area and assess the damage severity, a fast and accurate loss assessment method is of great importance for effective management. This study, t...

  13. Bioenergy Promise Versus a Bug: How Different Cultivars of Switchgrass Fare Against the Fall Armyworm (Spodoptera frugiperda)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Switchgrass (Panicum virgatum L.) is a North American native plant that is being considered for bioenergy production. A number of different commercial cultivars were evaluated for resistance to the fall armyworm. Differential feeding resistance was detected among the cultivars, but no insect morta...

  14. Phylogenetic molecular species delimitations unravel potential new species in the pest genus Spodoptera Guenée, 1852 (Lepidoptera, Noctuidae).

    PubMed

    Dumas, Pascaline; Barbut, Jérôme; Le Ru, Bruno; Silvain, Jean-François; Clamens, Anne-Laure; d'Alençon, Emmanuelle; Kergoat, Gael J

    2015-01-01

    Nowadays molecular species delimitation methods promote the identification of species boundaries within complex taxonomic groups by adopting innovative species concepts and theories (e.g. branching patterns, coalescence). As some of them can efficiently deal with large single-locus datasets, they could speed up the process of species discovery compared to more time consuming molecular methods, and benefit from the existence of large public datasets; these methods can also particularly favour scientific research and actions dealing with threatened or economically important taxa. In this study we aim to investigate and clarify the status of economically important moths species belonging to the genus Spodoptera (Lepidoptera, Noctuidae), a complex group in which previous phylogenetic analyses and integrative approaches already suggested the possible occurrence of cryptic species and taxonomic ambiguities. In this work, the effectiveness of innovative (and faster) species delimitation approaches to infer putative species boundaries has been successfully tested in Spodoptera, by processing the most comprehensive dataset (in terms of number of species and specimens) ever achieved; results are congruent and reliable, irrespective of the set of parameters and phylogenetic models applied. Our analyses confirm the existence of three potential new species clusters (for S. exigua (Hübner, 1808), S. frugiperda (J.E. Smith, 1797) and S. mauritia (Boisduval, 1833)) and support the synonymy of S. marima (Schaus, 1904) with S. ornithogalli (Guenée, 1852). They also highlight the ambiguity of the status of S. cosmiodes (Walker, 1858) and S. descoinsi Lalanne-Cassou & Silvain, 1994. This case study highlights the interest of molecular species delimitation methods as valuable tools for species discovery and to emphasize taxonomic ambiguities. PMID:25853412

  15. Phylogenetic Molecular Species Delimitations Unravel Potential New Species in the Pest Genus Spodoptera Guenée, 1852 (Lepidoptera, Noctuidae)

    PubMed Central

    Dumas, Pascaline; Barbut, Jérôme; Le Ru, Bruno; Silvain, Jean-François; Clamens, Anne-Laure; d’Alençon, Emmanuelle; Kergoat, Gael J.

    2015-01-01

    Nowadays molecular species delimitation methods promote the identification of species boundaries within complex taxonomic groups by adopting innovative species concepts and theories (e.g. branching patterns, coalescence). As some of them can efficiently deal with large single-locus datasets, they could speed up the process of species discovery compared to more time consuming molecular methods, and benefit from the existence of large public datasets; these methods can also particularly favour scientific research and actions dealing with threatened or economically important taxa. In this study we aim to investigate and clarify the status of economically important moths species belonging to the genus Spodoptera (Lepidoptera, Noctuidae), a complex group in which previous phylogenetic analyses and integrative approaches already suggested the possible occurrence of cryptic species and taxonomic ambiguities. In this work, the effectiveness of innovative (and faster) species delimitation approaches to infer putative species boundaries has been successfully tested in Spodoptera, by processing the most comprehensive dataset (in terms of number of species and specimens) ever achieved; results are congruent and reliable, irrespective of the set of parameters and phylogenetic models applied. Our analyses confirm the existence of three potential new species clusters (for S. exigua (Hübner, 1808), S. frugiperda (J.E. Smith, 1797) and S. mauritia (Boisduval, 1833)) and support the synonymy of S. marima (Schaus, 1904) with S. ornithogalli (Guenée, 1852). They also highlight the ambiguity of the status of S. cosmiodes (Walker, 1858) and S. descoinsi Lalanne-Cassou & Silvain, 1994. This case study highlights the interest of molecular species delimitation methods as valuable tools for species discovery and to emphasize taxonomic ambiguities. PMID:25853412

  16. Data set for diet specific differential gene expression analysis in three Spodoptera moths.

    PubMed

    Roy, A; Walker, W B; Vogel, H; Kushwaha, S K; Chattington, S; Larsson, M C; Anderson, P; Heckel, D G; Schlyter, F

    2016-09-01

    Examination of closely related species pairs is suggested for evolutionary comparisons of different degrees of polyphagy, which we did here with three taxa of lepidopteran herbivores, Spodoptera spp (S. littoralis, S. frugiperda maize (C) and rice (R) strains) for a RNAseq analysis of the midguts from the 3rd instar insect larvae for differential metabolic responses after feeding on pinto bean based artificial diet vs maize leaves. Paired-end (2×100 bp) Illumina HiSeq2500 sequencing resulted in a total of 24, 23, 24, and 21 million reads for the SF-C-Maize, SF-C-Pinto, SF-R-Maize, SF-R Pinto, and a total of 35 and 36 million reads for the SL-Maize and SL-Pinto samples, respectively. After quality control measures, a total of 62.2 million reads from SL and 71.7 million reads from SF were used for transcriptome assembly (TA). The resulting final de novo reference TA (backbone) for the SF taxa contained 37,985 contigs with a N50 contig size of 1030 bp and a maximum contig length of 17,093 bp, while for SL, 28,329 contigs were generated with a N50 contig size of 1980 bp and a maximum contig length of 18,267 bp. The data presented herein contains supporting information related to our research article Roy et al. (2016) http://dx.doi.org/10.1016/j.ibmb.2016.02.006[1]. PMID:27366783

  17. Monitoring for exotic Spodoptera species (Lepidoptera: Noctuidae) in Florida

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Trapping studies were conducted in two Florida locations to determine if three Old World Spodoptera Guenée species were present. Commercially-produced lures for S. exempta (Walker), S. littoralis (Boisduval), and S. litura (F.), plus a S. litura lure made by the USDA-APHIS-CPHST laboratory at Otis ...

  18. [Characterization of the damage of Spodoptera eridania (Cramer) and Spodoptera cosmioides (Walker) (Lepidoptera: Noctuidae) to structures of cotton plants].

    PubMed

    Santos, Karen B Dos; Meneguim, Ana M; Santos, Walter J Dos; Neves, Pedro M O J; Santos, Rachel B Dos

    2010-01-01

    The cotton plant, Gossypium hirsutum, hosts various pests that damage different structures. Among these pests, Spodoptera cosmioides (Walker) and Spodoptera eridania (Cramer) (Lepidoptera: Noctuidae) are considered important. The objectives of this study were to characterize and to quantify the potential damage of S. eridania and S. cosmioides feeding on different structures of cotton plants. For this purpose, newly-hatched larvae were reared on the following plant parts: leaf and flower bud; leaf and boll; flower bud or boll; and leaf, flower bud and boll. The survival of S. cosmioides and S. eridania was greater than 80% and 70% for larvae fed on cotton plant parts offered separately or together, respectively. One larva of S. eridania damaged 1.7 flower buds, but did not damage bolls, while one larva of S. cosmioides damaged 5.2 flower buds and 3.0 cotton bolls. Spodoptera eridania and S. cosmioides can be considered species with potential to cause economic damage to cotton plants because they can occur throughout cotton developmental stages causing defoliation and losses of reproductive structures. Therefore, the results validate field observations that these two species of Spodoptera are potential pests for cotton. PMID:20878002

  19. The role of the PI3K-Akt signal transduction pathway in Autographa californica multiple nucleopolyhedrovirus infection of Spodoptera frugiperda cells

    SciTech Connect

    Xiao Wei; Yang Yi; Weng Qingbei; Lin Tiehao; Yuan Meijin; Yang Kai; Pang Yi

    2009-08-15

    Many viruses activate the phosphatidylinositol 3-kinase (PI3K)-Akt signaling pathway, thereby modulating diverse downstream signaling pathways associated with antiapoptosis, proliferation, cell cycling, protein synthesis and glucose metabolism, in order to augment their replication. To date, the role of the PI3K-Akt pathway in Baculovirus replication has not been defined. In the present study, we demonstrate that infection of Sf9 cells with Autographa californica multiple nucleopolyhedrovirus (AcMNPV) elevated cellular Akt phosphorylation at 1 h post-infection. The maximum Akt phosphorylation occurred at 6 h post-infection and remained unchanged until 18 h post-infection. The PI3K-specific inhibitor, LY294002, suppressed Akt phosphorylation in a dose-dependent manner, suggesting that AcMNPV-induced Akt phosphorylation is PI3K-dependent. The inhibition of PI3K-Akt activation by LY294002 significantly reduced the viral yield, including a reduction in budded viruses and occlusion bodies. The virus production was reduced only when the inhibitor was added within 24 h of infection, implying that activation of PI3K occurred early in infection. Correspondingly, both viral DNA replication and late (VP39) and very late (POLH) viral protein expression were impaired by LY294002 treatment; LY294002 had no effect on immediate-early (IE1) and early-late (GP64) protein expression. These results demonstrate that the PI3K-Akt pathway is required for efficient Baculovirus replication.

  20. Production and purification of human papillomavirus type 33 L1 virus-like particles from Spodoptera frugiperda 9 cells using two-step column chromatography.

    PubMed

    Baek, Jin-Oh; Seo, Jeong-Woo; Kim, Ik-Hwan; Kim, Chul Ho

    2011-02-01

    The major capsid protein L1 of human papillomavirus (HPV) is essential in construction of recombinant antigen vaccines against cervical cancer. HPV type 33 accounts for about 10% of all HPV infections in Asia. The gene encoding the major capsid protein L1 of the high-risk HPV type 33 was isolated from a Korean patient and expressed in Sf-9 insect cells using a baculovirus expression system. HPV33 L1 protein was isolated by two-step chromatographic purification using strong-cation exchange and ceramic hydroxyapatite chromatography. Strong-cation-exchange chromatography was performed to achieve initial purification of HPV33 L1 and to remove most contaminating proteins, and secondary ceramic hydroxyapatite chromatography yielded pure HPV33 L1 virus-like particles (VLPs). Ceramic hydroxyapatite columns are particularly useful in the purification of antibodies, antigens, human viruses, and VLPs, and we thus used this system. The expression of HPV L1 protein in Sf-9 cells was examined by SDS-PAGE, Western-blotting, and ELISA analyses, and the data showed that HPV33 L1 VLPs were determined to > 98% purity and 58.7% recovery by a quantitative immuno-ELISA assay. Transmission electron microscopy analysis revealed that the HPV VLPs were approximately 50-60 nm in diameter and created by self-assembly of HPV L1 protein. The efficient and simple purification process described here should be useful in production of a cervical cancer vaccine. PMID:20716445

  1. Determination and analysis of the genome sequence of Spodoptera littoralis multiple nucleopolyhedrovirus

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Spodoptera littoralis multiple nucleopolyhedrovirus (SpliMNPV), a pathogen of the Egyptian cotton leaf worm Spodoptera littoralis, was subjected to sequencing of its entire DNA genome and bioassay analysis comparing its virulence to that of other baculoviruses. The annotated SpliMNPV genome of...

  2. A cardioactive peptide from the southern armyworm, Spodoptera eridania.

    PubMed

    Furuya, K; Hackett, M; Cirelli, M A; Schegg, K M; Wang, H; Shabanowitz, J; Hunt, D F; Schooley, D A

    1999-01-01

    A cardioactive peptide was isolated from extracts of whole heads of the southern armyworm, Spodoptera eridania. This peptide has the sequence ENFAVGCTPGYQRTADGRCKPTF (Mr = 2516.8), determined from both Edman sequencing and tandem mass spectrometry in combination with off-line micropreparative capillary liquid chromatography. This peptide, termed Spoer-CAP23, has excitatory effects on a semi-isolated heart from larval Manduca sexta, causing an inotropic effect at low concentrations of peptide and chronotropic and inotropic effects at high doses. The threshold concentration for stimulatory effects of the synthetic peptide on the semi-isolated heart was about 1 nM, suggesting a physiological role as a neuropeptide. PMID:10098624

  3. Antifeedant activity of Momordica dioica fruit pulp extracts on Spodoptera litura.

    PubMed

    Narasimhan, S; Kannan, S; Ilango, K; Maharajan, G

    2005-12-01

    The hexane extract and ethyl acetate soluble fraction of methanolic extract of the fruit pulp of Momordica dioica exhibited moderate and concentration dependent antifeedant activity against Spodoptera litura. PMID:16246499

  4. Susceptibility, mechanisms of response and resistance to Bacillus thuringiensis toxins in Spodoptera spp.

    PubMed

    Herrero, Salvador; Bel, Yolanda; Hernández-Martínez, Patricia; Ferré, Juan

    2016-06-01

    Bioinsecticides based on Bacillus thuringiensis have long been used as an alternative to synthetic insecticides to control insect pests. In this review, we focus on insects of the genus Spodoptera, including relevant polyphagous species that are primary and secondary pests of many crops, and how B. thuringiensis toxins can be used for Spodoptera spp. pest management. We summarize the main findings related to susceptibility, midgut binding specificity, mechanisms of response and resistance of this insect genus to B. thuringiensis toxins. PMID:27436737

  5. Identification and comparative expression analysis of odorant binding protein genes in the tobacco cutworm Spodoptera litura.

    PubMed

    Gu, Shao-Hua; Zhou, Jing-Jiang; Gao, Shang; Wang, Da-Hai; Li, Xian-Chun; Guo, Yu-Yuan; Zhang, Yong-Jun

    2015-01-01

    Insect odorant binding proteins (OBPs) are thought to involve in insects' olfaction perception. In the present study, we identified 38 OBP genes from the antennal transcriptomes of Spodoptera litura. Tissue expression profiles analysis revealed that 17 of the 38 SlitOBP transcripts were uniquely or primarily expressed in the antennae of both sexes, suggesting their putative role in chemoreception. The RPKM value analysis revealed that seven OBPs (SlitPBP1-3, SlitGOBP1-2, SlitOBP3 and SlitOBP5) are highly abundant in male and female antennae. Most S. litura antennal unigenes had high homology with Lepidoptera insects, especially genes of the genus Spodoptera. Phylogenetic analysis of the Lepidoptera OBPs demonstrated that the OBP genes from the genus Spodoptera (S. litura, Spodoptera littoralis and Spodoptera exigua) had a relatively close evolutionary relationship. Some regular patterns and key conserved motifs of OBPs in genus Spodoptera are identified by MEME, and their putative roles in detecting odorants are discussed here. The motif-patterns between Lepidoptera OBPs and CSPs are also compared. The SlitOBPs identified here provide a starting point to facilitate functional studies of insect OBPs at the molecular level both in vivo and in vitro. PMID:26346731

  6. Identification and comparative expression analysis of odorant binding protein genes in the tobacco cutworm Spodoptera litura

    PubMed Central

    Gu, Shao-Hua; Zhou, Jing-Jiang; Gao, Shang; Wang, Da-Hai; Li, Xian-Chun; Guo, Yu-Yuan; Zhang, Yong-Jun

    2015-01-01

    Insect odorant binding proteins (OBPs) are thought to involve in insects’ olfaction perception. In the present study, we identified 38 OBP genes from the antennal transcriptomes of Spodoptera litura. Tissue expression profiles analysis revealed that 17 of the 38 SlitOBP transcripts were uniquely or primarily expressed in the antennae of both sexes, suggesting their putative role in chemoreception. The RPKM value analysis revealed that seven OBPs (SlitPBP1-3, SlitGOBP1-2, SlitOBP3 and SlitOBP5) are highly abundant in male and female antennae. Most S. litura antennal unigenes had high homology with Lepidoptera insects, especially genes of the genus Spodoptera. Phylogenetic analysis of the Lepidoptera OBPs demonstrated that the OBP genes from the genus Spodoptera (S. litura, Spodoptera littoralis and Spodoptera exigua) had a relatively close evolutionary relationship. Some regular patterns and key conserved motifs of OBPs in genus Spodoptera are identified by MEME, and their putative roles in detecting odorants are discussed here. The motif-patterns between Lepidoptera OBPs and CSPs are also compared. The SlitOBPs identified here provide a starting point to facilitate functional studies of insect OBPs at the molecular level both in vivo and in vitro. PMID:26346731

  7. Immature Development of Spodoptera dolichos (Fabricius) (Lepidoptera: Noctuidae).

    PubMed

    Montezano, D G; Sosa-Gómez, D R; Paula-Moraes, S V; Roque-Specht, V F; Fronza, E; Barros, N M; Specht, A

    2016-02-01

    We provide detailed temporal and morphological parameters of the immature stages of Spodoptera dolichos (Fabricius) larvae fed on artificial diet under controlled conditions (25 ± 1 °C, 70 ± 10% RH, and 14 h photophase). The viability of the egg, larval, pupal, and prepupal stages was 97.5%, 97.0%, 93.1%, and 98.9%, respectively. The average duration of the egg, larval, prepupal, and pupal stages was 5.0, 23.4, 3.2, and 21.5 days, respectively. Females took longer at the larval stage than males, with 10.5% of them having seven instars. The growth rate of female larvae that developed through six and seven instars was 1.72 and 1.54, respectively. Female pupae were significantly larger, exhibiting slower development than males. PMID:26429580

  8. Using DNA barcoding and genitalia to place “Leucochlaena” hipparis (Druce) in Spodoptera Guenee (Lepidoptera: Noctuidae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Using the 658 bp ‘barcode’ region of cytochrome oxidase (COI) gene, the species “Leucochlaena” hipparis (Druce) was placed in the genus Spodoptera. A neighbor-joining tree placing “Leucochlaena” hipparis within Spodoptera is illustrated. Adult and male and female genitalia are illustrated....

  9. Use of DNA Barcodes to Identify Invasive Armyworm Spodoptera Species in Florida

    PubMed Central

    Nagoshi, Rodney N.; Brambila, Julieta; Meagher, Robert L.

    2011-01-01

    A critical component for sustaining adequate food production is the protection of local agriculture from invasive pest insects. Essential to this goal is the ability to accurately distinguish foreign from closely related domestic species, a process that has traditionally required identification using diagnostic morphological “keys” that can be both subtle and labor-intensive. This is the case for the Lepidopteran group of insects represented by Spodoptera, a genus of Noctuidae “armyworm” moths that includes several important agricultural pests. Two of the most destructive species, Spodoptera littoralis (Boisduval) (Lepidoptera: Noctuidae) and S. litura (F.) are not yet established in North America. To facilitate the monitoring for these pests, the feasibility of using DNA barcoding methodology for distinguishing between domestic and foreign Spodoptera species was tested. A DNA barcoding database was derived for a subset of Spodoptera species native to Florida, with an emphasis on those attracted to pheromone blends developed for S. litura or S. littoralis. These were then compared to the barcode sequences of S. litura collected from Taiwan and S. littoralis from Portugal. Consistent discrimination of the different species was obtained with phenetic relationships produced that were generally in agreement with phylogenetic studies using morphological characteristics. The data presented here indicate that DNA barcoding has the potential to be an efficient and accurate supplement to morphological methods for the identification of invasive Spodoptera pests in North America. PMID:22239735

  10. Pupal melanization is associated with higher fitness in Spodoptera exigua

    PubMed Central

    Liu, Sisi; Wang, Mo; Li, Xianchun

    2015-01-01

    Melanism has long been thought to be a habitat adaptation with a fitness cost. Here we reported a homozygous melanic strain (SEM) of Spodoptera exigua (Hübner) (Insecta: Lepidoptera: Noctuidae) established with black pupae spontaneously occurring within a typical laboratory population (SEW). The melanization is expressed globally, and only in the pupal stage. After pupation, the melanic SEM pupae gradually accumulate melanin to become completely black within 6 hours, whereas the wild-type SEW pupae gradually turn yellow-brown. The melanic SEM strain exhibits faster development in all life stages, heavier pupa weight, more mating time, higher fecundity, and accordingly, higher net reproductive rate and population trend index. While no reproductive isolation was observed between the SEM and SEW strains, the mating times per female of the reciprocal crosses and the SEM intracrosses were significantly higher than those of the SEW intracrosses. This represents a rare case of melanization that has fitness gains, rather than costs. Analysis of the life-history traits of this case and 14 previously reported cases of insect melanism indicate that none of melanization origin, stage, space and variation type determining whether melanism will cause fitness gain or cost. PMID:26039886

  11. Pupal melanization is associated with higher fitness in Spodoptera exigua.

    PubMed

    Liu, Sisi; Wang, Mo; Li, Xianchun

    2015-01-01

    Melanism has long been thought to be a habitat adaptation with a fitness cost. Here we reported a homozygous melanic strain (SEM) of Spodoptera exigua (Hübner) (Insecta: Lepidoptera: Noctuidae) established with black pupae spontaneously occurring within a typical laboratory population (SEW). The melanization is expressed globally, and only in the pupal stage. After pupation, the melanic SEM pupae gradually accumulate melanin to become completely black within 6 hours, whereas the wild-type SEW pupae gradually turn yellow-brown. The melanic SEM strain exhibits faster development in all life stages, heavier pupa weight, more mating time, higher fecundity, and accordingly, higher net reproductive rate and population trend index. While no reproductive isolation was observed between the SEM and SEW strains, the mating times per female of the reciprocal crosses and the SEM intracrosses were significantly higher than those of the SEW intracrosses. This represents a rare case of melanization that has fitness gains, rather than costs. Analysis of the life-history traits of this case and 14 previously reported cases of insect melanism indicate that none of melanization origin, stage, space and variation type determining whether melanism will cause fitness gain or cost. PMID:26039886

  12. Enhanced resistance to Spodoptera litura in endophyte infected cauliflower plants.

    PubMed

    Thakur, Abhinay; Kaur, Sanehdeep; Kaur, Amarjeet; Singh, Varinder

    2013-04-01

    Endophytic fungi, which live within host plant tissues without causing any visible symptom of disease, are important mediators of plant-herbivore interactions. These endophytes enhance resistance of host plant against insect herbivores mainly by productions of various alkaloid based defensive compounds in the plant tissue or through alterations of plant nutritional quality. Two endophytic fungi, i.e., Nigrospora sp. and Cladosporium sp., were isolated from Tinospora cordifolia (Thunb.) Miers, a traditional indian medicinal plant. Cauliflower (Brassica oleracea L.) plants were inoculated with these two endophytic fungi. The effect of endophyte infected and uninfected cauliflower plants were measured on the survival and development of Spodoptera litura (Fab.), a polyphagous pest. Endophyte infected cauliflower plants showed resistance to S. litura in the form of significant increase in larval and pupal mortality in both the fungi. Inhibitory effects of endophytic fungi also were observed on adult emergence, longevity, reproductive potential, as well as hatchability of eggs. Thus, it is concluded that antibiosis to S. litura could be imparted by artificial inoculation of endophytes and this could be used to develop alternative ecologically safe control strategies. PMID:23575013

  13. Laboratory evaluation of Isaria fumosorosea against Spodoptera littoralis.

    PubMed

    Zemek, R; Hussein, H M; Prenerová, E

    2012-01-01

    Isaria fumosorosea (syn. Paecilomyces fumosoroseus) is potentially useful for the biological control of economically important agricultural and forest insect pests. We evaluated efficacy of two strains of this entomopathogenic fungus against last instar larvae and pupae of Egyptian cotton leafworm, Spodoptera littoralis. The first strain was Apopka 97 which is an active ingredient of commercial biopesticide PreFeRal WG (Biobest, Belgium). The second strain was isolated from Cameraria ohridella and is currently deposited under number CCM 8367 as a patent culture in the Czech Collection of Microorganisms in Brno. Blastospores of both strains were obtained after 120 hours submerged cultivation in grow media using orbital shaker. The concentration of blastospores was adjusted to 5 x 10(7) spores/ml of suspension. Soaking agent Tween 80 was added to the suspension at concentration 0.02%. Lethal effects of both fungal strains on S. littoralis were evaluated using standard dip test. Treated insects were individually placed into plastic Petri dishes (diam. 9 cm) and kept at constant laboratory conditions (temperature 23 degrees C, R.H. approx. 100%, 16L:8D photoperiod). Virulence of the strains was expressed as percentages of cumulative daily mortality corrected for mortality in the control variant. Obtained results revealed higher virulence of CCM 8367 blastospores to the last instar larvae of S. littoralis (93.1% mortality) on the 7th day after the treatment compared to Apopka 97 (65.5% mortality). Even more obvious difference was found in pupae, where corrected mortality of CCM 8367-treated pupae was 80.0% while mortality in Apopka 97-treated pupae reached only 3.3% on the 8th day after the treatment. We can conclude that the strain I. fumosorosea CCM 8367 has strong insecticidal effects on S. littoralis and has a potential to be implemented as a novel biocontrol agent. PMID:23885438

  14. Active role of fatty acid amino acid conjugates in nitrogen metabolidm by Spodoptera litura larvae

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Since the first fatty acid amino acid conjugate (FAC) was isolated from regurgitant of Spodoptera exigua larvae in 1997 [volicitin: N-(17-hydroxylinolenoyl)- L-glutamine], their role as elicitors of induced responses in plants has been well documented. However, studies of the biosyntheses as well as...

  15. Monitoring two native Spodoptera species using an exotic pheromone lure developed for an exotic species

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The pheromone lure for the exotic species Spodoptera exempta was successful at attracting two native species, S. latifascia and S. albula. Trapping was conducted in north-central Florida and in southern Texas. Large numbers of both native species were collected throughout the season....

  16. PGE2 induces oenocytoid cell lysis via a G protein-coupled receptor in the beet armyworm, Spodoptera exigua

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Eicosanoids mediate cellular and humoral immune responses in the beet armyworm, Spodoptera exigua, including activation of prophenoloxidase (PPO). PPO activation begins with release of its inactive zymogen, PPO, from oenocytoids in response to prostaglandins (PGs). Based on the biomedical literatur...

  17. Haplotype profile comparisons between Spodoptera frugiperda (Lepidoptera:Noctuidae) populations from Mexico with those from Puerto Rico, South America, and the United States and their implications for migratory behavior

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fall armyworm is a major economic pest throughout the Western Hemisphere of maize, cotton, sorghum, and a variety of agricultural grasses and vegetable crops. Previous studies demonstrated extensive annual migrations occurring as far north as Canada from overwintering locations in southern Florida a...

  18. Relation ofSpodoptera eridania choice to tannins and protein oflotus corniculatus.

    PubMed

    Briggs, M A

    1990-05-01

    Plant secondary compounds such as tannins may influence herbivore choice. To determine if herbivory was influenced by tannin concentration,Spodoptera eridania larvae were given a choice ofLotus corniculatus plants whose chemical profiles were altered by fertilization. Herbivores chose plants that had been grown with symbiotic nitrogen fixation as their only nitrogen source more often than fertilized plants. Choice was related to protein concentration, but not to tannin concentration. PMID:24263826

  19. Quantitative and ultrastructural changes in the haemocytes of Spodoptera littoralis (Boisd.) treated individually or in combination with Spodoptera littoralis multicapsid nucleopolyhedrovirus (SpliMNPV) and azadirachtin.

    PubMed

    Shaurub, El-Sayed H; Abd El-Meguid, Afaf; Abd El-Aziz, Nahla M

    2014-10-01

    The total haemocyte count (THC) and the possible ultrastructural alterations induced in the haemocytes of the fourth larval instars of the Egyptian cotton leafworm, Spodoptera littoralis (Boisd.) (Lepidoptera: Noctuidae), 96 h post-feeding on a semi-synthetic diet, treated with the LC50 of Spodoptera littoralis multicapsid nucleopolyhedrovirus (SpliMNPV) and the LC50 of azadirachtin alone, and the LC25 of SpliMNPV combined with the LC25 of azadirachtin were studied and compared to the control. Single treatment with the virus and azadirachtin or combined treatment significantly decreased the THC compared to the control. There are five types of haemocytes in S. littoralis: prohaemocytes, plasmatocytes, granulocytes, spherulocytes and oenocytoids. The most common symptoms in granulocytes and plasmatocytes, the main affected cell types, due to viral infection were the presence of virogenic stroma, peripheral dispersion of the chromatin and disappearance of the nucleoli. However, the most common symptoms in these two types of haemocytes due to treatment with azadirachtin were the presence of rough endoplasmic reticulum filled with fibrous materials, due to probably apoptosis, in their cisternae and disorganization of mitochondria (looped, vacuolated and swollen). In addition, the cytoplasm of granulocytes was vacuolated with the appearance of autophagic lysosomes, while plasmatocytes showed ruptured cell membrane and folded nuclear envelope. Combined treatment with the NPV and azadirachtin induced the same pathological changes which were recorded from individual treatment with the virus or azadirachtin to the same haemocytes. It can be concluded that the change in the THC and ultrastructure of granulocytes and plasmatocytes may affect the cellular-mediated immune response in S. littoralis. Moreover, it seems likely that mitochondria were the target site of azadirachtin, as they were affected in both granulocytes and plasmatocytes treated with azadirachtin alone or in

  20. Tropical soda apple (Solanum viarum) mediated competition via induced resistance: Interaction between Gratiana boliviana, Spodoptera exigua and Frankliniella occidentalis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Survival assays were conducted with beet armyworm (BAW) Spodoptera exigua (Hübner), a tortoise beetle Gratiana bolivana Spaeth and western flower thrips (WFT) Frankliniella occidentalis (Pergande) on tropical soda apple (TSA) Solanum viarum Dunal, a relative of tomato. Both S. exigua and G. bolivia...

  1. 40 CFR 180.1118 - Spodoptera exigua nuclear polyhedrosis virus; exemption from the requirement of a tolerance.

    Code of Federal Regulations, 2012 CFR

    2012-07-01

    ... virus; exemption from the requirement of a tolerance. 180.1118 Section 180.1118 Protection of... polyhedrosis virus; exemption from the requirement of a tolerance. An exemption from the requirement of a tolerance is established for the microbial pest control agent Spodoptera exigua nuclear polyhedrosis...

  2. 40 CFR 180.1118 - Spodoptera exigua nuclear polyhedrosis virus; exemption from the requirement of a tolerance.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... virus; exemption from the requirement of a tolerance. 180.1118 Section 180.1118 Protection of... polyhedrosis virus; exemption from the requirement of a tolerance. An exemption from the requirement of a tolerance is established for the microbial pest control agent Spodoptera exigua nuclear polyhedrosis...

  3. 40 CFR 180.1118 - Spodoptera exigua nuclear polyhedrosis virus; exemption from the requirement of a tolerance.

    Code of Federal Regulations, 2010 CFR

    2010-07-01

    ... virus; exemption from the requirement of a tolerance. 180.1118 Section 180.1118 Protection of... polyhedrosis virus; exemption from the requirement of a tolerance. An exemption from the requirement of a tolerance is established for the microbial pest control agent Spodoptera exigua nuclear polyhedrosis...

  4. 40 CFR 180.1118 - Spodoptera exigua nuclear polyhedrosis virus; exemption from the requirement of a tolerance.

    Code of Federal Regulations, 2011 CFR

    2011-07-01

    ... virus; exemption from the requirement of a tolerance. 180.1118 Section 180.1118 Protection of... polyhedrosis virus; exemption from the requirement of a tolerance. An exemption from the requirement of a tolerance is established for the microbial pest control agent Spodoptera exigua nuclear polyhedrosis...

  5. 40 CFR 180.1118 - Spodoptera exigua nuclear polyhedrosis virus; exemption from the requirement of a tolerance.

    Code of Federal Regulations, 2013 CFR

    2013-07-01

    ... virus; exemption from the requirement of a tolerance. 180.1118 Section 180.1118 Protection of... polyhedrosis virus; exemption from the requirement of a tolerance. An exemption from the requirement of a tolerance is established for the microbial pest control agent Spodoptera exigua nuclear polyhedrosis...

  6. Relationship of Flight and Reproduction in Beet Armyworm, Spodoptera exigua (Lepidoptera: Noctuidae), a Migrant Lacking the Oogenesis-flight Syndrome

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The beet armyworm, Spodoptera exigua, undertakes long-distance migration. We used flight mills to investigate the interaction between flight and reproduction in this species given the apparent absence of the oogenesis-flight syndrome. This syndrome is characterized by a suite of traits common in man...

  7. Nitrogen fertilization rate affects feeding, larval performance, and oviposition preference of the beet armyworm, Spodoptera exigua, on cotton

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Nitrogen (N) is one of the most critical chemical elements for plant and animal growth. Development and oviposition of the beet armyworm, Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae) was studied in relation to varying nitrogen levels in cotton, Gossypium hirsutum L. The development of S. exig...

  8. Target and non-target toxicity of botanical insecticide derived from Couroupita guianensis L. flower against generalist herbivore, Spodoptera litura Fab. and an earthworm, Eisenia foetida Savigny

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Novel chemistries in botanical insecticides may provide alternatives to, or development of synthetic insecticides suitable for controlling the Lepidopteran pests, like Spodoptera litura (F.). Many botanical chemistries are biodegradable, and have lower mammalian toxicity. Eight natural chemical comp...

  9. Complete sequence and gene organization of the Nosema spodopterae rRNA gene.

    PubMed

    Tsai, Shu-Jen; Huang, Wei-Fone; Wang, Chung-Hsiung

    2005-01-01

    By sequencing the entire ribosomal RNA (rRNA) gene of Nosema spodopterae, we show here that its gene organization follows a pattern similar to the Nosema type species, Nosema bombycis, i.e. 5'-large subunit rRNA (2,497 bp)-internal transcribed spacer (185 bp)-small subunit rRNA (1,232 bp)-intergenic spacer (277 bp)-5S rRNA (114 bp)-3'. Gene sequences and the secondary structures of large subunit rRNA, small subunit rRNA, and 5S rRNA are compared with the known corresponding sequences and structures of closely related microsporidia. The results suggest that the Nosema genus may be heterogeneous and that the rRNA gene organization may be a useful characteristic for determining which species are closely related to the type species. PMID:15702980

  10. Isolation and identification of a cardioactive peptide from Tenebrio molitor and Spodoptera eridania.

    PubMed

    Furuya, K; Liao, S; Reynolds, S E; Ota, R B; Hackett, M; Schooley, D A

    1993-12-01

    We isolated several cardioactive peptides from extracts of whole heads of the mealworm, Tenebrio molitor, and the southern armyworm, Spodoptera eridania, using a semi-isolated heart of Manduca sexta for bioassay. We have now isolated from each species the peptide with the strongest effect on rate of contraction of the heart. The peptides were identified using micro Edman sequencing and mass spectrometric methods. This cardioactive peptide has the same primary structure from both species: Pro-Phe-Cys-Asn-Ala-Phe-Thr-Gly-Cys-NH2, a cyclic nonapeptide which is identical to crustacean cardioactive peptide (CCAP) originally isolated from the shore crab, Carcinus maenas, and subsequently isolated from Locusta migratoria and Manduca sexta. This is additional evidence that CCAP has widespread occurrence in arthropoda. PMID:8129851

  11. Mitomycin C induced alterations in antioxidant enzyme levels in a model insect species, Spodoptera eridania.

    PubMed

    Batcabe, J P; MacGill, R S; Zaman, K; Ahmad, S; Pardini, R S

    1994-05-01

    1. An insect species, the southern armyworm Spodoptera eridania, was used as an in vivo model to examine mitomycin C's (MMC) pro-oxidant effect reflected in alterations of antioxidant enzymes. 2. Following a 2-day exposure to 0.01 and 0.05% w/w dietary concentrations, MMC only induced superoxide dismutase activity. All other enzyme activities were not affected, indicating oxidative stress was mild. 3. Following a 5-day exposure to 0.05% w/w dietary MMC, the activities of superoxide dismutase, glutathione-S-transferase and its peroxidase activity and DT-diaphorase were induced. GR activity was not altered. The high constitutive catalase activity was also not affected. These responses of S. eridania's antioxidant enzymes are analogous to those of mammalian systems in alleviating MMC-induced oxidative stress. 4. S. eridania emerges as an appropriate non-mammalian model for initial and cost-effective screening of drug-induced oxidative stress. PMID:7926607

  12. Bioefficacy of Aristolochia tagala Cham. against Spodoptera litura Fab. (Lepidoptera: Noctuidae)

    PubMed Central

    Baskar, K.; Sasikumar, S.; Muthu, C.; Kingsley, S.; Ignacimuthu, S.

    2010-01-01

    Bioefficacy of leaf and root extracts of Aristolochia tagala Cham. at different concentrations was evaluated at room temperature against Spodoptera litura Fab. Effects on feeding, larvicidal and pupicidal activities and larval–pupal duration were studied. Higher antifeedant activity (56.06%), lethal concentration for feeding inhibition (3.69%), larvicidal (40.66%), pupicidal (28%), total mortality (68.66%) and prolonged larval–pupal duration (12.04–13.08 days) were observed in ethyl acetate leaf extract at 5.0% concentration. Dose dependant effect of test extracts was observed. This plant could be used to isolate active principles and to develop a new botanical formulation in pest management programmes. PMID:23961100

  13. Complete mitochondrial genome of the common cutworm Spodoptera litura (Lepidoptera: Noctuidae).

    PubMed

    Li, Feng-Bo; Zhang, Hui-Xian; Shen, Wei-Feng; Xu, Xiao-Yan; Wang, Wei; Chen, Jin-E; He, Li-Hua; Meng, Zhi-Qi

    2015-01-01

    We determined the complete mitochondrial genome of the common cutworm Spodoptera litura (Lepidoptera: Noctuidae), which is one of the most destructive polyphagous insect pests worldwide. The genome is 15,383 bp in length (GenBank accession number: KF701043) with an A+T content of 81.08%, and contains 37 typical animal mitochondrial genes (13 protein-coding genes, 2 rRNA genes and 22 tRNA genes) with the typical arrangement found in Lepidoptera. All the protein-coding genes (PCGs) start with ATN start codon except for cox1, which begins with CGA. Eight PCGs stop with complete termination codons (TAA or TAG), whereas five PCGs use incomplete stop codon T. The A+T-rich region is located between rrnS and trnM with a length of 326 bp and an A+T content of 93.87%, and harbors three tandem repeat elements. PMID:24409845

  14. Genomic Sequence Analysis of Granulovirus Isolated from the Tobacco Cutworm, Spodoptera litura

    PubMed Central

    Wang, Yong; Choi, Jae Young; Roh, Jong Yul; Liu, Qin; Tao, Xue Ying; Park, Jong Bin; Kim, Jae Su; Je, Yeon Ho

    2011-01-01

    Background Spodoptera litura is a noctuid moth that is considered an agricultural pest. The larvae feed on a wide range of plants and have been recorded on plants from 40 plant families (mostly dicotyledons). It is a major pest of many crops. To better understand Spodoptera litura granulovirus (SpliGV), the nucleotide sequence of the SpliGV DNA genome was determined and analyzed. Methodology/Principal Findings The genome of the SpliGV was completely sequenced. The nucleotide sequence of the SpliGV genome was 124,121 bp long with 61.2% A+T content and contained 133 putative open reading frames (ORFs) of 150 or more nucleotides. The 133 putative ORFs covered 86.3% of the genome. Among these, 31 ORFs were conserved in most completely sequenced baculovirus genomes, 38 were granulovirus (GV)-specific, and 64 were present in some nucleopolyhedroviruses (NPVs) and/or GVs. We proved that 9 of the ORFs were SpliGV specific. Conclusions/Significance The genome of SpliGV is 124,121 bp in size. One hundred thirty-three ORFs that putatively encode proteins of 50 or more amino acid residues with minimal overlap were determined. No chitinase or cathepsin genes, which are involved in the liquefaction of the infected host, were found in the SpliGV genome, explaining why SpliGV-infected insects do not degrade in a typical manner. The DNA photolyase gene was first found in the genus Granulovirus. When phylogenic relationships were analyzed, the SpliGV was most closely related to Trichoplusia ni granulovirus (TnGV) and Xestia c-nigrum granulovirus (XecnGV), which belong to the Type I-granuloviruses (Type I-GV). PMID:22132235

  15. Baculovirus-Induced Climbing Behavior Favors Intraspecific Necrophagy and Efficient Disease Transmission in Spodoptera exigua

    PubMed Central

    Rebolledo, Dulce; Guevara, Roger; Murillo, Rosa

    2015-01-01

    Shortly prior to death, many species of Lepidoptera infected with nucleopolyhedrovirus climb upwards on the host plant. This results in improved dissemination of viral occlusion bodies over plant foliage and an increased probability of transmission to healthy conspecific larvae. Following applications of Spodoptera exigua multiple nucleopolyhedrovirus for control of Spodoptera exigua on greenhouse-grown sweet pepper crops, necrophagy was observed by healthy S. exigua larvae that fed on virus-killed conspecifics. We examined whether this risky behavior was induced by olfactory or phagostimulant compounds associated with infected cadavers. Laboratory choice tests and olfactometer studies, involving infected and non-infected cadavers placed on spinach leaf discs, revealed no evidence for greater attraction of healthy larvae to virus-killed over non-infected cadavers. Physical contact or feeding on infected cadavers resulted in a very high incidence of transmission (82–93% lethal disease). Observations on the behavior of S. exigua larvae on pepper plants revealed that infected insects died on the uppermost 10% of foliage and closer to the plant stem than healthy conspecifics of the same stage, which we considered clear evidence of baculovirus-induced climbing behavior. Healthy larvae that subsequently foraged on the plant were more frequently observed closer to the infected than the non-infected cadaver. Healthy larvae also encountered and fed on infected cadavers significantly more frequently and more rapidly than larvae that fed on non-infected cadavers. Intraspecific necrophagy on infected cadavers invariably resulted in virus transmission and death of the necrophagous insect. We conclude that, in addition to improving the dissemination of virus particles over plant foliage, baculovirus-induced climbing behavior increases the incidence of intraspecific necrophagy in S. exigua, which is the most efficient mechanism of transmission of this lethal pathogen. PMID

  16. Baculovirus-Induced Climbing Behavior Favors Intraspecific Necrophagy and Efficient Disease Transmission in Spodoptera exigua.

    PubMed

    Rebolledo, Dulce; Lasa, Rodrigo; Guevara, Roger; Murillo, Rosa; Williams, Trevor

    2015-01-01

    Shortly prior to death, many species of Lepidoptera infected with nucleopolyhedrovirus climb upwards on the host plant. This results in improved dissemination of viral occlusion bodies over plant foliage and an increased probability of transmission to healthy conspecific larvae. Following applications of Spodoptera exigua multiple nucleopolyhedrovirus for control of Spodoptera exigua on greenhouse-grown sweet pepper crops, necrophagy was observed by healthy S. exigua larvae that fed on virus-killed conspecifics. We examined whether this risky behavior was induced by olfactory or phagostimulant compounds associated with infected cadavers. Laboratory choice tests and olfactometer studies, involving infected and non-infected cadavers placed on spinach leaf discs, revealed no evidence for greater attraction of healthy larvae to virus-killed over non-infected cadavers. Physical contact or feeding on infected cadavers resulted in a very high incidence of transmission (82-93% lethal disease). Observations on the behavior of S. exigua larvae on pepper plants revealed that infected insects died on the uppermost 10% of foliage and closer to the plant stem than healthy conspecifics of the same stage, which we considered clear evidence of baculovirus-induced climbing behavior. Healthy larvae that subsequently foraged on the plant were more frequently observed closer to the infected than the non-infected cadaver. Healthy larvae also encountered and fed on infected cadavers significantly more frequently and more rapidly than larvae that fed on non-infected cadavers. Intraspecific necrophagy on infected cadavers invariably resulted in virus transmission and death of the necrophagous insect. We conclude that, in addition to improving the dissemination of virus particles over plant foliage, baculovirus-induced climbing behavior increases the incidence of intraspecific necrophagy in S. exigua, which is the most efficient mechanism of transmission of this lethal pathogen. PMID

  17. Transcriptome Analysis of the SL221 Cells at the Early Stage during Spodoptera litura Nucleopolyhedrovirus Infection

    PubMed Central

    Yu, Qian; Xiong, Youhua; Liu, Jianliang; Wen, Dongling; Wu, Xiaohui; Yin, Hanqi

    2016-01-01

    Spodoptera litura (S. litura) is one of the most destructive agricultural pests worldwide. There is urgent need for a nuclear polyhedrosis virus that is specific to S. litura. To date, there have been no reports regarding the responses of S. litura cells to early Spodoptera litura nucleopolyhedrovirus (SpltNPV) infection due to the lack of a reference genome and transcriptome for S. litura. In this study, a cell transcriptome from the host S. litura was assembled and used for Illumina strand-specific RNA sequencing (RNA-seq) to generate 99180 unigenes, representing the 18 hour infection cycle. More than 2000 S. litura genes were significant differentially regulated throughout the infection. The levels of viral mRNAs began to increase dramatically at 6 hpi, and this increase continued throughout the remainder of the infection. We focused on the expression of genes related to stress responses, apoptosis, metabolic enzymes and host cell innate immune system. A small subset of genes related to host stress response, especially for 62 ones being able to annotated as enzyme, ligand and receptor genes, were observed to be specifically differentially expressed at 6 hpi. At 18 hpi, 104 unigenes were continuously significantly changing from 0 hpi to 18 hpi, considered to be viral multiplication related genes, including 3 annotated SL221 unigenes and 81 viral genes, such as tetraspanin and iap gene. This information and further studies on the regulation of host gene expression by baculovirus infection at early stage will provide the tools needed to enhance the utility of this virus as an effective insecticide. PMID:26840182

  18. Endophyte-mediated interactions between cauliflower, the herbivore Spodoptera litura, and the ectoparasitoid Bracon hebetor.

    PubMed

    Kaur, Tamanreet; Singh, Bahaderjeet; Kaur, Amarjeet; Kaur, Sanehdeep

    2015-10-01

    Fungal endosymbionts in plants may influence interactions among plants, herbivores and their parasitoids through the production of secondary metabolites. We used a lepidopteran pest and its generalist parasitoid to test the effect of endophyte-infected plants on a third trophic level. Endophytic fungi, Aspergillus flavus and Aspergillus niger, isolated from Acacia arabica, were used to infect cauliflower plants. We found that the presence of the endophyte in the plants significantly extended the development period of Spodoptera litura (Fab.) larvae. Feeding of the host on endophyte-infected plants further adversely affected the development and performance of its parasitoid, Bracon hebetor (Say). A negative impact was also recorded for longevity and fecundity of endophyte-naive parasitoid females due to the parasitization of host larvae fed on endophyte-infected plants. The presence of endophytes in the diet of the host larvae significantly prolonged the development of the parasitoid. A strong detrimental effect was also recorded for larval survival and emergence of parasitoid adults. The longevity and parasitism rate of female wasps were reduced significantly due to the ingestion of endophyte-infected cauliflower plants by S. litura larvae. Overall, we found that both endophytic fungi had a negative impact on the parasitoid. PMID:26041060

  19. Ovicidal activity of Atalantia monophylla (L) Correa against Spodoptera litura Fab. (Lepidoptera: Noctuidae)

    PubMed Central

    Baskar, Kathirvelu; Muthu, Chellaiah; Raj, Gnanaprakasam Antony; Kingsley, Selvadurai; Ignacimuthu, Savarimuthu

    2012-01-01

    Objective To evaluate the efficacy of Atalantia monophylla (A. monophylla) leaf in different solvent crude extracts and fractions against eggs of Spodoptera litura (S. litura). Methods Hexane, ethyl acetate and chloroform solvent extracts of A. monophylla leaf and 12 fractions from hexane extract were screened at 5.0%, 2.5%, 1.0% and 0.5% for crude extracts and 1 000, 500, 250 and 125 mg/kg for fractions against the eggs of S. litura for the ovicidal activity. LC50 and LC90 were calculated using probit analysis. Results Hexane crude extract showed maximum ovicidal activity of 61.94% at 5.0% concentration with a correlation value of r2=0.81, and least LC50 value of 3.06%. Hexane extract was fractionated using silica gel column chromatography and 12 fractions were obtained. Fraction 9 was active which showed maximum ovicidal activity of 75.61% at 1 000 mg/kg with the LC50 value of 318.65 mg/kg and LC90 value of 1 473.31 mg/kg. In linear regression analysis, significant and high correlation (r2=0.81%) was seen between concentration and ovicidal activity of hexane crude extracts and its active fraction. Conclusions As per our knowledge, this is the first report for ovicidal activity of A. monophylla against S. litura, A. monophylla could be used for the management of S. litura and other insect pests. PMID:23593580

  20. Impact of Elevated CO2 on Tobacco Caterpillar, Spodoptera litura on Peanut, Arachis hypogea

    PubMed Central

    Srinivasa Rao, M; Manimanjari, D; Vanaja, M; Rama Rao, CA; Srinivas, K; Rao, Vum; Venkateswarlu, B

    2012-01-01

    If the carbon dioxide (CO2) concentration in the atmosphere changes in the future, as predicted, it could influence crops and insect pests. The growth and development of the tobacco caterpillar, Spodoptera litura (Fabricius) (Noctuidae: Lepidoptera), reared on peanut (Arachis hypogea L.) foliage grown under elevated CO2 (550 ppm and 700 ppm) concentrations in open top chambers at Central Research Institute for Dryland Agriculture, Hyderabad, India, were examined in this study. Significantly lower leaf nitrogen, higher carbon, higher relative proportion of carbon to nitrogen and higher polyphenols content expressed in terms of tannic acid equivalents were observed in the peanut foliage grown under elevated CO2 levels. Substantial influence of elevated CO2 on S. litura was noticed, such as longer larval duration, higher larval weights, and increased consumption of peanut foliage by S. litura larvae under elevated CO2 compared with ambient CO2. Relative consumption rate was significantly higher for S. litura larva fed plants grown at 550 and 700 ppm than for larvae fed plants grown at ambient condition. Decreased efficiency of conversion of ingested food, decreased efficiency of conversion of digested food, and decreased relative growth rate of larvae was observed under elevated CO2. The present results indicate that elevated CO2 levels altered the quality of the peanut foliage, resulting in higher consumption, lower digestive efficiency, slower growth, and longer time to pupation (one day more than ambient). PMID:23437971

  1. Bioinsecticidal activity of Murraya koenigii miraculin-like protein against Helicoverpa armigera and Spodoptera litura.

    PubMed

    Gahloth, Deepankar; Shukla, Umesh; Birah, Ajanta; Gupta, Gorakh P; Kumar, P Ananda; Dhaliwal, Harcharan S; Sharma, Ashwani K

    2011-11-01

    Miraculin-like proteins, belonging to the Kunitz superfamily, are natural plant defense agents against pests and predators, and therefore are potential biopesticides for incorporation into pest-resistant crops. Here, a miraculin-like protein from Murraya koenigii was assessed for its in vitro and in vivo effects against two polyphagous lepidopteran insect pests, Helicoverpa armigera and Spodoptera litura. M. koenigii miraculin-like protein (MKMLP) inhibited the trypsin-like activity and total protease activity of H. armigera gut proteinases (HGP) by 78.5 and 40%, respectively, and S.litura gut proteinases (SGP) by 81 and 48%, respectively. The inhibitor was stable and actively inhibited the proteolysis of both HGP and SGP enzymes for up to 72 h. Incorporation of MKMLP into artificial diet adversely affected the growth and development of pests in a dose-dependent manner. After 10 days of feeding on diets containing 200 µM MKMLP, larval weight was reduced to 69 and 44.8% and larval mortality was increased to 40 and 43.3% for H. armigera and S litura, respectively. The LC(50) of MKMLP was 0.34 and 0.22% of the diet for H.armigera and S. litura, respectively. These results demonstrate the efficacy of MKMLP as a potential plant defense agent against H. armigera and S. litura. PMID:21948662

  2. An antennal circadian clock and circadian rhythms in peripheral pheromone reception in the moth Spodoptera littoralis.

    PubMed

    Merlin, Christine; Lucas, Philippe; Rochat, Didier; François, Marie-Christine; Maïbèche-Coisne, Martine; Jacquin-Joly, Emmanuelle

    2007-12-01

    Circadian rhythms are observed in mating behaviors in moths: females emit sex pheromones and males are attracted by these pheromones in rhythmic fashions. In the moth Spodoptera littoralis, we demonstrated the occurrence of a circadian oscillator in the antenna, the peripheral olfactory organ. We identified different clock genes, period (per), cryptochrome1 (cry1) and cryptochrome2 (cry2), in this organ. Using quantitative real-time PCR (qPCR), we found that their corresponding transcripts cycled circadianly in the antenna as well as in the brain. Electroantennogram (EAG) recordings over 24 h demonstrated for the first time a circadian rhythm in antennal responses of a moth to sex pheromone. qPCR showed that out of one pheromone-binding protein (PBP), one olfactory receptor (OR), and one odorant-degrading enzyme (ODE), all putatively involved in the pheromone reception, only the ODE transcript presented a circadian rhythm that may be related to rhythms in olfactory signal resolution. Peripheral or central circadian clock control of olfaction is then discussed in light of recent data. PMID:18057325

  3. CRISPR/Cas9-mediated targeted gene mutagenesis in Spodoptera litura.

    PubMed

    Bi, Hong-Lun; Xu, Jun; Tan, An-Jiang; Huang, Yong-Ping

    2016-06-01

    Custom-designed nuclease technologies such as the clustered regularly interspaced short palindromic repeat (CRISPR)-associated (Cas) system provide attractive genome editing tools for insect functional genetics. The targeted gene mutagenesis mediated by the CRISPR/Cas9 system has been achieved in several insect orders including Diptera, Lepidoptera and Coleoptera. However, little success has been reported in agricultural pests due to the lack of genomic information and embryonic microinjection techniques in these insect species. Here we report that the CRISPR/Cas9 system induced efficient gene mutagenesis in an important Lepidopteran pest Spodoptera litura. We targeted the S. litura Abdominal-A (Slabd-A) gene which is an important embryonic development gene and plays a significant role in determining the identities of the abdominal segments of insects. Direct injection of Cas9 messenger RNA and Slabd-A-specific single guide RNA (sgRNA) into S. litura embryos successfully induced the typical abd-A deficient phenotype, which shows anomalous segmentation and ectopic pigmentation during the larval stage. A polymerase chain reaction-based analysis revealed that the Cas9/sgRNA complex effectively induced a targeted mutagenesis in S. litura. These results demonstrate that the CRISPR/Cas9 system is a powerful tool for genome manipulation in Lepidopteran pests such as S. litura. PMID:27061764

  4. Development and Leaf Consumption by Spodoptera cosmioides (Walker) (Lepidoptera: Noctuidae) Reared on Leaves of Agroenergy Crops.

    PubMed

    Cabezas, M F; Nava, D E; Geissler, L O; Melo, M; Garcia, M S; Krüger, R

    2013-12-01

    Spodoptera cosmioides (Walker) (Lepidoptera: Noctuidae) is a polyphagous pest that threatens more than 24 species of crop plants including those used for biodiesel production such as Ricinus communis (castor bean), Jatropha curcas (Barbados nut), and Aleurites fordii (tung oil tree). The development and leaf consumption by S. cosmioides reared on leaves of these three species were studied under controlled laboratory conditions. The egg-to-adult development time of S. cosmioides was shortest when reared on castor bean leaves and longest when reared on tung oil tree leaves. Larvae reared on castor bean and Barbados nut leaves had seven instars, whereas those reared on tung oil tree leaves had eight. Females originating from larvae reared on castor bean and Barbados nut leaves showed greater fecundity than did females originating from larvae reared on tung oil tree leaves. Insects fed on castor bean leaves had shorter life spans than those fed on tung oil tree and Barbados nut leaves although the oviposition period did not differ significantly. The intrinsic and finite rates of increase were highest for females reared on castor bean leaves. Total leaf consumption was highest for larvae reared on tung oil tree leaves and lowest for those reared on Barbados nut leaves. We conclude that castor bean is a more appropriate host plant for the development of S. cosmioides than are Barbados nut and tung oil tree. PMID:27193276

  5. Antioxidant enzyme level response to prooxidant allelochemicals in larvae of the southern armyworm moth, Spodoptera eridania.

    PubMed

    Pritsos, C A; Ahmad, S; Elliott, A J; Pardini, R S

    1990-01-01

    Larvae of the southern armyworm, Spodoptera eridania, are highly polyphagous feeders which frequently encounter and feed upon plants containing high levels of prooxidant allelochemicals. While ingestion of moderate quantities of prooxidants can be tolerated by these larvae, ingestion of larger quantities can result in toxicity. Studies were conducted to assess the role of the antioxidant enzymes, superoxide dismutase (SOD), catalase (CAT) and glutathione reductase (GR) in the protection of S. eridania against redox active prooxidant plant allelochemicals. Dietary exposure of mid-fifth-instar larvae to either quercetin (a flavonoid) or xanthotoxin (a photoactive furanocoumarin), which generate superoxide radical, and singlet oxygen, respectively, resulted in an increase in SOD levels. CAT levels increased in all groups of S. eridania including control insects. This may have been due to the sudden exposure to food following an extended fast of 18 h (to insure that larvae would not reject the diet because of the prooxidants' bitter taste) with an eventual lowering of CAT values with time. GR activities did not significantly change except for a slight inhibition at the highest prooxidant concentrations used at 12-h post-ingestion. The data from these studies suggest that SOD responds to prooxidant challenges in these insects and together with CAT and GR contributes to the insect's defense against potentially toxic prooxidant compounds. PMID:2161387

  6. Identification of volatile sex pheromone components released by the southern armyworm,Spodoptera eridania (Cramer).

    PubMed

    Teal, P E; Mitchell, E R; Tumlinson, J H; Heath, R R; Sugie, H

    1985-06-01

    Analysis of sex pheromone gland extracts and volatile pheromone components collected from the calling female southern armyworm,Spodoptera eridania (Cramer), by high-resolution capillary gas chromatography and mass spectroscopy indicated that a number of 14-carbon mono- and diunsaturated acetates and a monounsaturated 16-carbon acetate were produced. Gland extracts also indicated the presence of (Z)-9-tetradecen-1-ol. However, this compound was not found in collections of volatiles. Field trapping studies indicated that the volatile blend composed of (Z)-9-tetradecen-1-ol acetate (60%), (Z)-9-(E)-12-tetradecadien-1-ol acetate (17%), (Z)-9-(Z)-12-tetradecadien-1-ol acetate (15%), (Z)-9-(E)-11-tetradecadien-1-ol acetate (5%), and (Z)-11-hexadecen-1-ol acetate (3 %) was an effective trap bait for males of this species. The addition of (Z)-9-tetradecen-1-ol to the acetate blends tested resulted in the capture of beet armyworm,S. exigua (Hubner), males which provides further evidence that the alcohol is a pheromone component of this species. PMID:24310218

  7. Dichlone-induced oxidative stress in a model insect species, Spodoptera eridania.

    PubMed

    Ahmad, S; Zaman, K; MacGill, R S; Batcabe, J P; Pardini, R S

    1995-11-01

    Southern armyworm, Spodoptera eridania, larvae were provided ad libitum 0.002-0.25% w/w dichlone, 2,3-dichloro-1,4-naphthoquinone (CNQ). Larval mortality occurred in a time-and-dose dependent manner, with an LC17 of 0.01% and an LC50 of 0.26% CNQ at day-5. Extracts of larvae fed control, 0.01, and 0.25% CNQ diets for 5 days were assayed for antioxidant enzymes. While 0.01% CNQ had a mild effect, 0.25% CNQ profoundly increased levels of all antioxidant enzymes that were examined. The increases as compared to control were: 5.3-, 1.9-, 3.2-, 2.6-, 2.8-, and 3.5-fold higher for superoxide dismutase, catalase, glutathione transferase and its peroxidase activity, glutathione reductase and DT-diaphorase, respectively. At 0.01% CNQ, the thiobarbituric acid reactive substances (TBARS) were similar to the control group. However, despite the induction from 0.25% CNQ of all enzymes examined, the lipid peroxidation was not attenuated; the TBARS were 29.7% over the control value. High mortalities and CNQ-induced pathologies reflected in retarded growth, wasting syndrome, and diuresis clearly indicated that the insect sustained severe oxidant-induced injuries before appropriate defenses were fully mobilized. Thus, this quinone causes an oxidative stress in a model insect species analogous to that observed in mammalian species. PMID:7574883

  8. Laboratory evaluation of Artemisia annua L. extract and artemisinin activity against Epilachna paenulata and Spodoptera eridania.

    PubMed

    Maggi, María E; Mangeaud, Arnaldo; Carpinella, María C; Ferrayoli, Carlos G; Valladares, Graciela R; Palacios, Sara M

    2005-07-01

    Ethanolic extract of aerial parts of Artemisia annua L. and artemisinin were evaluated as anti-insect products. In a feeding deterrence assay on Epilachna paenulata Germ (Coleoptera: Coccinellidae) larvae, complete feeding rejection was observed at an extract concentration of 1.5 mg/cm2 on pumpkin leaf tissue. The same concentration produced a feeding inhibition of 87% in Spodoptera eridania (Cramer) (Lepidoptera: Noctuidae). In a no-choice assay, both species ate less and gained less weight when fed on leaves treated with the extract. Complete mortality in E. paenulata and 50% mortality in S. eridania were observed with extract at 1.5 mg/cm2. Artemisinin exhibited a moderate antifeedant effect on E. paenulata and S. eridania at 0.03-0.375 mg/cm2. However, a strong effect on survival and body weight was observed when E. paenulata larvae were forced to feed on leaves treated at 0.03 and 0.075 mg/cm2. Artemisia annua ethanolic extract of aerial parts at 1.5 mg/cm2 showed no phytotoxic effect on pumpkin seedlings. PMID:16222790

  9. Immature stages of Spodoptera eridania (Lepidoptera: Noctuidae): developmental parameters and host plants.

    PubMed

    Montezano, Débora Goulart; Specht, Alexandre; Sosa-Gómez, Daniel Ricardo; Roque-Specht, Vânia Ferreira; de Barros, Neiva Monteiro

    2014-01-01

    This study aimed to detail the temporal and morphological parameters of the immature stages of southern armyworm Spodoptera eridania (Stoll, 1782) with larvae feed on artificial diet, under controlled conditions (25 ± 1°C, 70 ± 10% relative humidity and 14-h photophase) and gather information about their larval host plants. The viability of the egg, larval, pupal, and prepupal stages was 97.82, 93.62, 96.42, and 97.03%, respectively. The average duration of the egg, larval, pupal, and pre-pupal stages was 4.00, 16.18, 1.58, and 9.17 d, respectively. During the larval stage, 43.44% of females passed through seven instars, observing that the female's development was significant slower than males. The female larvae that developed through six and seven instars exhibited a mean growth rate of 1.52 and 1.44, respectively. Female pupae were significantly larger, exhibiting faster development than males. The rearing method proved to be adequate, providing more detailed observations of the biological cycle, especially at the larval stage, and resulting in an overall survival of almost 85%. Two hundred two plant species belonging to 58 families are listed as natural hosts for S. eridania, mainly including Asteraceae, Fabaceae, Solanaceae, Poaceae, Amaranthaceae, and Malvaceae. PMID:25525103

  10. 3'-phosphoadenosine-5'-phosphosulphate synthesis and involvement in sulphotransferase reactions in the insect, Spodoptera littoralis.

    PubMed Central

    Isaac, R E; Phua, K K; Rees, H H

    1982-01-01

    1. Synthesis of 3'-phosphoadenosine-5'-phosphosulphate from ATP and 35SO4(-2) was demonstrated by homogenates of gut. Malpighian tubules and fat body of Spodoptera littoralis. 2. The enzyme system was most active in the gut tissue, and was primarily located in the cytosol fraction of the cell. Gut cytosol preparations were used as a source of the 3'-phosphoadenosine-5'-phosphosulphate generating system for more detailed studies. 3. Maximum synthesis required an incubation mixture containing Tris/HCl buffer (pH 7.5), ATP (20 mM), MgCl2 (13.0 mM) and K2SO4 (3 mM). 4. The specific activity of 3'-phosphoadenosine-5'-phosphosulphate synthesizing activity in gut cytosol increased during development of the sixth instar larva, reaching a peak at day 4. A sudden fall in specific activity was observed in the prepupal stage. 5. 3'-Phosphoadenosine-5'-phosphosulphate formation is the rate limiting process in the overall sulphation of p-nitrophenol in the gut cytosol preparations from S. littoralis. 6. It is concluded that the properties of the sulphate-activating system in this insect are similar to those reported for vertebrates. PMID:6956335

  11. Antibiosis in Soybean Genotypes and the Resistance Levels to Spodoptera eridania (Cramer) (Lepidoptera: Noctuidae).

    PubMed

    Souza, B H S; Silva, A G; Janini, J C; Boica Júnior, A L

    2014-12-01

    The southern armyworm (SAW) Spodoptera eridania (Cramer) is one of the most common armyworm species defoliating soybeans. Preliminary screening trials have indicated that some soybean genotypes exhibit resistance to SAW. Therefore, in this study, we evaluated the development of SAW larvae fed on ten soybean genotypes in order to identify genotypes with antibiosis-type resistance. Neonate SAW larvae were daily fed with young leaves collected from plants at the vegetative growth stages V4-V5. Larval development and survival were recorded. Genotypes PI 227687 and PI 227682 delayed larval, pupal, and larva-adult development and yielded larvae with the lowest weight and survival and pupae with the lowest weight. Genotypes IAC 100 and DM 339 also negatively affected larval and pupal development and larval survival but at a lower level. Based on our results, the soybean lines PI 227687 and PI 227682 could be used as sources of genes for soybean breeding programs aiming to develop high yield, SAW-resistant cultivars. Moreover, further trials must be carried out under field conditions to validate if the commercial cultivars IAC 100 and DM 339, which expressed moderate levels of antibiosis-type resistance in the laboratory, are effective in suppressing SAW larvae populations. PMID:27194067

  12. Overexpression of Tyrosine hydroxylase and Dopa decarboxylase associated with pupal melanization in Spodoptera exigua

    PubMed Central

    Liu, Sisi; Wang, Mo; Li, Xianchun

    2015-01-01

    Melanism has been found in a wide range of species, but the molecular mechanisms involved remain largely elusive. In this study, we studied the molecular mechanisms of the pupal melanism in Spodoptera exigua. The full length cDNA sequences of tyrosine hydroxylase (TH) and dopa decarboxylase (DDC), two key enzymes in the biosynthesis pathway of melanin, were cloned, and their temporal expression patterns in the integument were compared during the larval-pupal metamorphosis process of the S. exigua wild type (SEW) and melanic mutant (SEM) strains. No amino acid change in the protein sequence of TH and DDC was found between the two strains. Both DDC and TH were significantly over-expressed in the integument of the SEM strain at late-prepupa and 0 h pupa, respectively, compared with those of the SEW strain. Feeding 5th instar larvae of SEM with diets incorporated with 1 mg/g of the DDC inhibitor L-α-Methyl-DOPA and 0.75 mg/g of the TH inhibitor 3-iodo-tyrosine (3-IT) resulted in 20% pupae with partially-rescued phenotype and 68.2% of pupae with partially- or fully-rescued phenotype, respectively. These results indicate that overexpressions of TH and DDC are involved in the pupal melanization of S. exigua. PMID:26084938

  13. Mechanism of entomotoxicity of the plant lectin from Hippeastrum hybrid (Amaryllis) in Spodoptera littoralis larvae.

    PubMed

    Caccia, Silvia; Van Damme, Els J M; De Vos, Winnok H; Smagghe, Guy

    2012-09-01

    Plant lectins have received a lot of attention because of their insecticidal properties. When orally administered in artificial diet or in transgenic plants, lectins provoke a wide range of detrimental effects, including alteration of the digestive enzyme machinery, fecundity drop, reduced feeding, changes in oviposition behavior, growth and development inhibition and mortality. Although many studies reported the entomotoxicity of lectins, only a few of them investigated the mode of action by which lectins exert toxicity. In the present paper we have studied for the first time the insecticidal potential of the plant lectin from Hippeastrum hybrid (Amaryllis) (HHA) bulbs against the larvae of the cotton leafworm (Spodoptera littoralis). Bioassays on neonate larvae showed that this mannose-specific lectin affected larval growth, causing a development retardation and larval weight decrease. Using primary cell cultures from S. littoralis midguts and confocal microscopy we have elucidated FITC-HHA binding and internalization mechanisms. We found that HHA did not exert a toxic effect on S. littoralis midgut cells, but HHA interaction with the brush border of midgut cells interfered with normal nutrient absorption in the S. littoralis midgut, thereby affecting normal larval growth in vivo. This study thus confirms the potential of mannose-specific lectins as pest control agents and sheds light on the mechanism underlying lectin entomotoxicity. PMID:22677323

  14. Microbial control of the cotton leafworm Spodoptera littoralis (Boisd.) by Egyptian Bacillus thuringiensis isolates.

    PubMed

    Alfazairy, Ahlam A; El-Ahwany, Amani M D; Mohamed, Eman A; Zaghloul, Heba A H; El-Helow, Ehab R

    2013-03-01

    Four local Bacillus thuringiensis (Bt) isolates that had been serologically identified as Bt var. kurstaki (Btk2, Btk3, and Btk66) and Bt var. mexicanensis (Btm27), in addition to two reference strains (4D20 and 4AC1), were laboratory assayed as microbial control agents against the Egyptian cotton leafworm Spodoptera littoralis (Boisd.). Polymerase chain reaction (PCR) amplification analysis revealed that each of the six experimental strains carries, at least, a cry1 type gene which expresses a protein toxin active against lepidopterous insects. Additionally, PCR amplification results demonstrated that 4D20 and Btk66 contain the Lepidoptera- and Diptera-active cry2 type gene and that Btk66 contains Coleoptera-active cry7 and cry8 genes. Among the six strains, Btk66 and Btm27 were the most promising microbial control agents against S. littoralis. The present findings were the first to report that Btm27 (classified as B. thuringiensis var. mexicanensis) is a very potent microbial control agent against S. littoralis-tested larvae. For more characterization of these two isolates, the sspO gene was investigated as a molecular chronometer. The DNA sequencing results proved that Btk66 and Btm27 carry sspO open reading frames with identical nucleotide sequences, suggesting a strong phylogenetic relationship between the two strains. PMID:22983675

  15. Roles of Peroxinectin in PGE2-Mediated Cellular Immunity in Spodoptera exigua

    PubMed Central

    Park, Jiyeong; Stanley, David; Kim, Yonggyun

    2014-01-01

    Background Prostaglandins (PGs) mediate insect immune responses to infections and invasions. Although the presence of PGs has been confirmed in several insect species, their biosynthesis in insects remains a conundrum because orthologs of the mammalian cyclooxygenases (COXs) have not been found in the known insect genomes. PG-mediated immune reactions have been documented in the beet armyworm, Spodoptera exigua. The purpose of this research is to identify the source of PGs in S. exigua. Principal Findings Peroxidases (POXs) are a sister group of COX genes. Ten putative POXs (SePOX-A ∼ SePOX-J) were expressed in S. exigua. Expressions of SePOX-F and -H were induced by bacterial challenge and expressed in the hemocytes and the fat body. RNAi of each POX was performed by hemocoelic injection of their specific double-stranded RNAs. dsPOX-F or, separately, dsPOX-H, but not the other eight dsRNA constructs, specifically suppressed hemocyte-spreading behavior and nodule formation; these two reactions were also inhibited by aspirin, a COX inhibitor. PGE2, but not arachidonic acid, treatment rescued the immunosuppression. Sequence analysis indicated that both POX genes were clustered with peroxinectin (Pxt) and their cognate proteins shared some conserved domains corresponding to the Pxt of Drosophila melanogaster. Conclusions SePOX-F and -H are Pxt-like genes associated with PG biosynthesis in S. exigua. PMID:25191834

  16. Effect of ace inhibitors and TMOF on growth, development, and trypsin activity of larval Spodoptera littoralis.

    PubMed

    Lemeire, Els; Borovsky, Dov; Van Camp, John; Smagghe, Guy

    2008-12-01

    Angiotensin converting enzyme (ACE) is a zinc metallopeptidase capable of cleaving dipeptide or dipeptideamide moieties at the C-terminal end of peptides. ACE is present in the hemolymph and reproductive tissues of insects. The presence of ACE in the hemolymph and its broad substrate specificity suggests an important role in processing of bioactive peptides. This study reports the effects of ACE inhibitors on larval growth in the cotton leafworm Spodoptera littoralis. Feeding ACE inhibitors ad lib decreased the growth rate, inhibited ACE activity in the larval hemolymph, and down-regulated trypsin activity in the larval gut. These results indicate that S. littoralis ACE may influence trypsin biosynthesis in the larval gut by interacting with a trypsin-modulating oostatic factor (TMOF). Injecting third instar larvae with a combination of Aea-TMOF and the ACE inhibitor captopril, down-regulated trypsin biosynthesis in the larval gut indicating that an Aea-TMOF gut receptor analogue could be present. Injecting captopril and enalapril into newly molted fifth instar larvae stopped larval feeding and decreased weight gain. Together, these results indicate that ACE inhibitors are efficacious in stunting larval growth and ACE plays an important role in larval growth and development. PMID:18949805

  17. Effects of perchlorate bioaccumulation on Spodoptera litura growth and sex ratio.

    PubMed

    Qin, Junhao; Shu, Yinghua; Li, Yongjun; He, Hongzhi; Li, Huashou

    2016-05-01

    Perchlorate (ClO4 (-)) pollution is widespread in the natural environment, but the effects of ClO4 (-) on the terrestrial insects are rarely studied. Here, when phytophagous insect Spodoptera litura larvae were fed on the diets with different ClO4 (-) concentrations, changes in their life-history traits were recorded; ClO4 (-) accumulations in feces and insect body were detected. The results demonstrated that ClO4 (-) bioaccumulation in insect at the different developmental stages was ranked in the order: adults > pupae > the 4th > 5th > 6th instar larvae. Besides, ClO4 (-) accumulations in the feces were ranked in the order: the 6th > 5th > 4th instar larvae. The ClO4 (-) accumulations in female pupae and adults were significantly higher than that in males. ClO4 (-) bioaccumulation in insect prolonged larval development time and caused a skewed sex ratio (the percentage of males at metamorphosis significantly decreased) under 100 to 200 mg ClO4 (-)/kg treatment. Therefore, ClO4 (-) accumulations in S. litura body presented developmental stage-, sex-specific pattern, and the sex-specific ClO4 (-) accumulations resulted in difference of sex ratio. These effects were observed at concentrations reported in natural environments contaminated with ClO4 (-), suggesting that this contaminant may pose a threat to the normal development and growth of this insect species. PMID:26810791

  18. TTAGG-repeat telomeres and characterization of telomerase in the beet armyworm, Spodoptera exigua (Lepidoptera: Noctuidae).

    PubMed

    Gong, H; Zhu, W; Zhang, J; Li, X; Meng, Q; Zhou, G; Wang, M; Wang, H; Miao, L; Qin, Q; Zhang, H

    2015-06-01

    Telomeres are maintained usually by telomerase, a specialized reverse transcriptase that adds this sequence to chromosome ends. In this study, telomerase activity was detected in the in different somatic tissues, such as midgut and fat bodies, by the telomeric repeat amplification protocol (TRAP) in Spodoptera exigua. The structure of the telomeres of S. exigua was evaluated by sequence analysis of the TRAP products, revealing that the telomerase synthesized a (TTAGG)n repeat. The presence of a telomerase reverse transcriptase (TERT) subunit coding gene has been cloned, sequenced and expressed in vitro successively. Notably, the S. exigua telomerase (SpexTERT) gene structure lacks the N-terminal GQ motif. Telomerase contains a large RNA subunit, TER, and a protein catalytic subunit, TERT. Here we report an in vitro system that was reconstructed by all components of the telomerase complex, a purified recombinant SpexTERT without a N-terminal GQ motif and a mutant human telomerase RNA (TER), showed telomerase activity. Together, these results suggest the GQ motif is not essential for telomerase catalysis. PMID:25689229

  19. Synthetic pheromones and plant volatiles alter the expression of chemosensory genes in Spodoptera exigua.

    PubMed

    Wan, Xinlong; Qian, Kai; Du, Yongjun

    2015-01-01

    Pheromone and plant odorants are important for insect mating, foraging food sources and oviposition. To understand the molecular mechanisms regulating pheromone and odorant signaling, we employed qRT-PCR to study the circadian rhythms of ABP, OBP, PBP, and OR gene expression in the beet armyworm, Spodoptera exigua and their responses after a pre-exposure to sex pheromone compounds or plant volatiles. The neuronal responses of male S. exigua to 20 chemical compounds were recorded at three specific time periods using the electroantennogram. The results showed a circadian rhythm in the expression profiles of some chemosensory genes in the antennae similar to their behavioral rhythm. The expression profiles of OR3, OR6, OR11, OR13, OR16, OR18, Orco, ABP2, OBP1, OBP7, and PBP1, and EAG responses to chemical compounds, as well as their circadian rhythm were significantly affected after exposure to synthetic sex pheromones and plant volatiles. These findings provide the first evidence that the gene expression of chemosensory genes and olfactory sensitivity to sex pheromones are affected by pre-exposing insects to pheromone compounds and plant volatiles. It helps to understand the molecular mechanisms underlying pheromone activity, and the application of sex pheromones and plant volatiles in mating disruption or mass trapping. PMID:26611815

  20. Overexpression of Tyrosine hydroxylase and Dopa decarboxylase associated with pupal melanization in Spodoptera exigua.

    PubMed

    Liu, Sisi; Wang, Mo; Li, Xianchun

    2015-01-01

    Melanism has been found in a wide range of species, but the molecular mechanisms involved remain largely elusive. In this study, we studied the molecular mechanisms of the pupal melanism in Spodoptera exigua. The full length cDNA sequences of tyrosine hydroxylase (TH) and dopa decarboxylase (DDC), two key enzymes in the biosynthesis pathway of melanin, were cloned, and their temporal expression patterns in the integument were compared during the larval-pupal metamorphosis process of the S. exigua wild type (SEW) and melanic mutant (SEM) strains. No amino acid change in the protein sequence of TH and DDC was found between the two strains. Both DDC and TH were significantly over-expressed in the integument of the SEM strain at late-prepupa and 0 h pupa, respectively, compared with those of the SEW strain. Feeding 5(th) instar larvae of SEM with diets incorporated with 1 mg/g of the DDC inhibitor L-α-Methyl-DOPA and 0.75 mg/g of the TH inhibitor 3-iodo-tyrosine (3-IT) resulted in 20% pupae with partially-rescued phenotype and 68.2% of pupae with partially- or fully-rescued phenotype, respectively. These results indicate that overexpressions of TH and DDC are involved in the pupal melanization of S. exigua. PMID:26084938

  1. Life history of Spodoptera exigua (Lepidoptera: Noctuidae) on various host plants.

    PubMed

    Azidah, A A; Sofian-Azirun, M

    2006-12-01

    The incubation period of Spodoptera exigua (Hübner) was not influenced by the host plant, whereas larval development time and pupal period were affected. Larval development time was longest on shallot and lady's finger, followed by cabbage and long bean. Larvae did not develop beyond the first instar when fed on chilli. The pupal period was longer on lady's finger than on cabbage, shallot and long bean. Overall, adult longevity was not influenced by the host plant but there was a difference between female and male longevity among the host plants. Survival of S. exigua was affected by the host plant at the larval stage. The number of larval instars varied between 5 and 8 within and between the studied host plants. Long bean was found to be the most suitable host plant and provide the best food quality for S. exigua compared to the other host plants, as it allowed faster development, fewer larval instars and a higher survival rate. PMID:17201979

  2. Identification and localization of two sensory neuron membrane proteins from Spodoptera litura (Lepidoptera: Noctuidae).

    PubMed

    Zhang, Jin; Liu, Yang; Walker, William B; Dong, Shuang-Lin; Wang, Gui-Rong

    2015-03-01

    Sensory neuron membrane proteins (SNMPs), which are located on the dendritic membrane of olfactory sensory neurons (OSNs), are proposed to be associated with odor reception in insects. Recent studies have demonstrated that SNMP1 is essential for electrophysiological responses of OSNs to the sex pheromone, cis-vaccenyl acetate (cVA) in Drosophila melanogaster. To investigate the function of Lepidoptera SNMPs, we cloned two SNMP genes, SlituSNMP1 and SltiuSNMP2, from Spodoptera litura (Lepidoptera: Noctuidae). Sequence alignment and phylogenetic analysis showed that both genes bear the general characteristics of SNMPs, including six conserved cysteine residues and two transmembrane domains. Further expression profile experiments showed that SlituSNMP1 is mainly expressed in the antenna, while SlituSNMP2 is broadly expressed in various tissues. By in situ hybridization experiments, it was found that SlituSNMP1 expressing cells are surrounded by the SlituSNMP2 expressing cells in the pheromone sensitive sensilla, suggesting different functions of the two SNMPs in insect olfaction. PMID:24757100

  3. Cryopreservation of the late stage embryos of Spodoptera exigua (Lepidoptera: Noctuidae).

    PubMed

    Luo, Li; Pang, Yi; Chen, Qijin; Li, Guanghong

    2006-01-01

    Genetic devolution, genetic drift and contamination are all threats to maintain germplasm stability during mass rearing of many insects. Cryopreservation of beet armworm (Spodoptera exigua) embryos was studied to provide information to improve mass rearing. A series of experiments was conducted on late-stage embryos (45-48 h at 27 degree C) of the beet armyworm, which included evaluation of cryoprotectants (CPAs), their toxicity and glass-forming tendency and optimization of experimental procedures. The results showed that ethylene glycol (EG) was the best CPA with comparatively low toxicity compared to the other six CPAs tested (methanol, 1,3-propanediol, glycerol, 2-amino-1-ethanol, 3-amino-1-propanol 3-methoxy-1 and 2-propanediol). The highest hatching rate of 8.8 degree was attained after freezing with a 3-step loading procedure and a 1-step unloading procedure, but the hatched larvae from frozen-thawed embryos did not actively feed and could not develop to a later stage. This was attributed to injuries from freezing in late stage embryos of S. exigua which had formed midguts. PMID:17256068

  4. Identification and Expression Profiles of Sex Pheromone Biosynthesis and Transport Related Genes in Spodoptera litura.

    PubMed

    Zhang, Ya-Nan; Zhu, Xiu-Yun; Fang, Li-Ping; He, Peng; Wang, Zhi-Qiang; Chen, Geng; Sun, Liang; Ye, Zhan-Feng; Deng, Dao-Gui; Li, Jin-Bu

    2015-01-01

    Although the general pathway of sex pheromone synthesis in moth species has been established, the molecular mechanisms remain poorly understood. The common cutworm Spodoptera litura is an important agricultural pest worldwide and causes huge economic losses annually. The female sex pheromone of S. litura comprises Z9,E11-14:OAc, Z9,E12-14:OAc, Z9-14:OAc, and E11-14:OAc. By sequencing and analyzing the transcriptomic data of the sex pheromone glands, we identified 94 candidate genes related to pheromone biosynthesis (55 genes) or chemoreception (39 genes). Gene expression patterns and phylogenetic analysis revealed that two desaturase genes (SlitDes5 and SlitDes11) and one fatty acyl reductase gene (SlitFAR3) showed pheromone gland (PG) biased or specific expression, and clustered with genes known to be involved in pheromone synthesis in other moth species. Furthermore, 4 chemoreception related genes (SlitOBP6, SlitOBP11, SlitCSP3, and SlitCSP14) also showed higher expression in the PG, and could be additional candidate genes involved in sex pheromone transport. This study provides the first solid background information that should facilitate further elucidation of sex pheromone biosynthesis and transport, and indicates potential targets to disrupt sexual communication in S. litura for a novel pest management strategy. PMID:26445454

  5. Identification and Expression Profiles of Sex Pheromone Biosynthesis and Transport Related Genes in Spodoptera litura

    PubMed Central

    Zhang, Ya-Nan; Zhu, Xiu-Yun; Fang, Li-Ping; He, Peng; Wang, Zhi-Qiang; Chen, Geng; Sun, Liang; Ye, Zhan-Feng; Deng, Dao-Gui; Li, Jin-Bu

    2015-01-01

    Although the general pathway of sex pheromone synthesis in moth species has been established, the molecular mechanisms remain poorly understood. The common cutworm Spodoptera litura is an important agricultural pest worldwide and causes huge economic losses annually. The female sex pheromone of S. litura comprises Z9,E11-14:OAc, Z9,E12-14:OAc, Z9-14:OAc, and E11-14:OAc. By sequencing and analyzing the transcriptomic data of the sex pheromone glands, we identified 94 candidate genes related to pheromone biosynthesis (55 genes) or chemoreception (39 genes). Gene expression patterns and phylogenetic analysis revealed that two desaturase genes (SlitDes5 and SlitDes11) and one fatty acyl reductase gene (SlitFAR3) showed pheromone gland (PG) biased or specific expression, and clustered with genes known to be involved in pheromone synthesis in other moth species. Furthermore, 4 chemoreception related genes (SlitOBP6, SlitOBP11, SlitCSP3, and SlitCSP14) also showed higher expression in the PG, and could be additional candidate genes involved in sex pheromone transport. This study provides the first solid background information that should facilitate further elucidation of sex pheromone biosynthesis and transport, and indicates potential targets to disrupt sexual communication in S. litura for a novel pest management strategy. PMID:26445454

  6. Virulence of Entomopathogenic Fungi Metarhizium anisopliae and Paecilomyces fumosoroseus for the Microbial Control of Spodoptera exigua

    PubMed Central

    Han, Ji Hee; Jin, Byung Rae; Lee, Sang Yeob

    2014-01-01

    The beet armyworm Spodoptera exigua (Lepidoptera: Noctuidae) is difficult to control using chemical insecticides because of the development of insecticide resistance. Several pest control agents are used to control the beet armyworm. Entomopathogenic fungi are one of the candidates for eco-friendly pest control instead of chemical control agents. In this study, among various entomopathogenic fungal strains isolated from soil two isolates were selected as high virulence pathogens against larva of beet armyworm. Control efficacy of fungal conidia was influenced by conidia concentration, temperature, and relative humidity (RH). The isolates Metarhizium anisopliae FT83 showed 100% cumulative mortality against second instar larvae of S. exigua 3 days after treatment at 1 × 107 conidia/mL and Paecilomyces fumosoroseus FG340 caused 100% mortality 6 days after treatment at 1 × 104 conidia/mL. Both M. anisopliae FT83 and P. fumosoroseus FG340 effectively controlled the moth at 20~30℃. M. anisopliae FT83 was significantly affected mortality by RH: mortality was 86.7% at 85% RH and 13.4% at 45% RH. P. fumosoroseus FG340 showed high mortality as 90% at 45% RH and 100% at 75% RH 6 days after conidia treatments. These results suggest that P. fumosoroseus FG340 and M. anisopliae FT83 have high potential to develop as a biocontrol agent against the beet armyworm. PMID:25606011

  7. Identification, Expression and Target Gene Analyses of MicroRNAs in Spodoptera litura

    PubMed Central

    Rao, Zhongchen; He, Wenyin; Liu, Lin; Zheng, Sichun; Huang, Lihua; Feng, Qili

    2012-01-01

    MicroRNAs (miRNAs) are small RNAs widely present in animals and plants and involved in post-transcriptional regulation of gene transcripts. In this study we identified and validated 58 miRNAs from an EST dataset of Spodoptera litura based on the computational and experimental analysis of sequence conservation and secondary structure of miRNA by comparing the miRNA sequences in the miRbase. RT-PCR was conducted to examine the expression of these miRNAs and stem-loop RT-PCR assay was performed to examine expression of 11 mature miRNAs (out of the 58 putative miRNA) that showed significant changes in different tissues and stages of the insect development. One hundred twenty eight possible target genes against the 11 miRNAs were predicted by using computational methods. Binding of one miRNA (sli-miR-928b) with the three possible target mRNAs was confirmed by Southern blotting, implying its possible function in regulation of the target genes. PMID:22662202

  8. Global and local modulatory supply to the mushroom bodies of the moth Spodoptera littoralis.

    PubMed

    Sinakevitch, Irina; Sjöholm, Marcus; Hansson, Bill S; Strausfeld, Nicholas J

    2008-07-01

    The moth Spodoptera littoralis, is a major pest of agriculture whose olfactory system is tuned to odorants emitted by host plants and conspecifics. As in other insects, the paired mushroom bodies are thought to play pivotal roles in behaviors that are elicited by contextual and multisensory signals, amongst which those of specific odors dominate. Compared with species that have elaborate behavioral repertoires, such as the honey bee Apis mellifera or the cockroach Periplaneta americana, the mushroom bodies of S. littoralis were originally viewed as having a simple cellular organization. This has been since challenged by observations of putative transmitters and neuromodulators. As revealed by immunocytology, the spodopteran mushroom bodies, like those of other taxa, are subdivided longitudinally into discrete neuropil domains. Such divisions are further supported by the present study, which also demonstrates discrete affinities to different mushroom body neuropils by antibodies raised against two putative transmitters, glutamate and gamma-aminobutyric acid, and against three putative neuromodulatory substances: serotonin, A-type allatostatin, and tachykinin-related peptides. The results suggest that in addition to longitudinal divisions of the lobes, circuits in the calyces and lobes are likely to be independently modulated. PMID:18406668

  9. Ultraviolet-B light induced oxidative stress: effects on antioxidant response of Spodoptera litura.

    PubMed

    Karthi, Sengodan; Sankari, R; Shivakumar, Muthugounder S

    2014-06-01

    Ultraviolet light (UV-B), which emits radiation in the range of 280-315 nm, has been used worldwide in light trapping of insect pests. In this article, we test the hypothesis that one of the duration of UV-B exposure has a differential impact on oxidative stress marker enzymes in Spodoptera litura. Effect of UV-B exposure on total protein and antioxidant activities of superoxide dismutase (SOD), catalase (CAT), peroxidases (POX) and glutathione-S-transferase (GST) were investigated in S. litura. The adults were exposed to UV-B light for various time periods (0, 30, 60, 90 and 120 min). We found that exposure to UV-B light for 30 and 60 min resulted in increased activities of POX. When the exposure time lasted for 60 and 90 min, the activities of SOD remained significantly higher than the control. However, the POX, CAT and GST activity decreased to control levels at 90 and 120 min. whereas relatively long duration exposure activates the xenobiotics detoxifying enzymes like GST and POX and CAT enzymes. Longer UV-B exposure may interfere with pesticide detoxification mechanism in insects, making them more susceptible to insecticides. PMID:24792567

  10. Synthetic pheromones and plant volatiles alter the expression of chemosensory genes in Spodoptera exigua

    PubMed Central

    Wan, Xinlong; Qian, Kai; Du, Yongjun

    2015-01-01

    Pheromone and plant odorants are important for insect mating, foraging food sources and oviposition. To understand the molecular mechanisms regulating pheromone and odorant signaling, we employed qRT-PCR to study the circadian rhythms of ABP, OBP, PBP, and OR gene expression in the beet armyworm, Spodoptera exigua and their responses after a pre-exposure to sex pheromone compounds or plant volatiles. The neuronal responses of male S. exigua to 20 chemical compounds were recorded at three specific time periods using the electroantennogram. The results showed a circadian rhythm in the expression profiles of some chemosensory genes in the antennae similar to their behavioral rhythm. The expression profiles of OR3, OR6, OR11, OR13, OR16, OR18, Orco, ABP2, OBP1, OBP7, and PBP1, and EAG responses to chemical compounds, as well as their circadian rhythm were significantly affected after exposure to synthetic sex pheromones and plant volatiles. These findings provide the first evidence that the gene expression of chemosensory genes and olfactory sensitivity to sex pheromones are affected by pre-exposing insects to pheromone compounds and plant volatiles. It helps to understand the molecular mechanisms underlying pheromone activity, and the application of sex pheromones and plant volatiles in mating disruption or mass trapping. PMID:26611815

  11. Insecticidal potential of an endophytic Cladosporium velox against Spodoptera litura mediated through inhibition of alpha glycosidases.

    PubMed

    Singh, Bahaderjeet; Kaur, Tamanreet; Kaur, Sanehdeep; Manhas, Rajesh K; Kaur, Amarjeet

    2016-07-01

    Alpha glucosidase inhibitory activity was exhibited by partially purified fractions obtained from an endophytic Cladosporium velox, isolated from Tinospora cordifolia. Taking into account the increasing importance of digestive enzyme inhibitors as insecticidal agents, the entomopathogenic potential of the fractions obtained was evaluated against Spodoptera litura (Fab.), a polyphagous pest. Considerable mortality was obtained when the larvae were fed on diet supplemented with the partially purified extract. All the concentrations of the extract significantly prolonged the overall developmental period of S. litura. At higher concentrations, the extract influenced the longevity of females as well as their reproductive potential. Phytochemical analysis revealed the presence of phenolic compounds in the active fraction. The phenolic compound responsible for the bioactivities was purified and identified to be chlorogenic acid using HPLC and MS analysis. The content of chlorogenic acid in the extract was quantified to be 250μg/ml. The purified compound also demonstrated inhibition of alpha glycosidases in vivo. The present study indicates that the endophyte imparted resistance to the insects in the plants could be mediated through chlorogenic acid targeting the alpha glycosidases present in the gut of the insect. The isolate obtained can be exploited for the production of chlorogenic acid, which has the potential to be exploited as a biocontrol agent against S. litura. PMID:27265826

  12. Resistance to selected organochlorin, organophosphate, carbamate and pyrethroid, in Spodoptera litura (Lepidoptera: Noctuidae) from Pakistan.

    PubMed

    Saleem, Mushtaq A; Ahmad, Munir; Ahmad, Mushtaq; Aslam, Muhammad; Sayyed, Ali H

    2008-10-01

    The toxicity of the most commonly used insecticides of organochlorine, organophosphate, pyrethroid, and carbamate groups were investigated against Spodoptera litura (F.) (Lepidoptera: Noctuidae) populations collected for three consecutive years (2004-2006). For a chlorocyclodiene and pyrethroids tested, the resistance ratios compared with Lab-PK were in the range of 10- to 92-fold for endosulfan, 5- to 111-fold for cypermethrin, 2- to 98-fold for deltamethrin, and 7- to 86-fold for beta-cyfluthrin. For organophosphates and carbamates, resistance ratios were in the range of 3- to 169-fold for profenofos, 18- to 421-fold for chlorpyrifos, 3- to 160-fold for quinalphos, 6- to 126-fold for phoxim, 7- to 463-fold for triazophos, and 10- to 389-fold for methomyl and 16- to 200-fold for thiodicarb. Resistance ratios were generally low to medium for deltamethrin and beta-cyfluthrin and high to very high for endosulfan, cypermethrin, profenofos, chlorpyrifos, quinalphos, phoxim, triazophos, methomyl, or thiodicarb. Pairwise comparisons of the log LC50 values of insecticides tested for all the populations showed correlations among several insecticides, suggesting a cross-resistance mechanism. Integration of timely judgment of pest problem, delimiting growing of alternate crops such as arum, rotation of insecticides with new chemicals, and insect growth regulators in relation to integrated pest management could help in manageable control of this important pest. PMID:18950050

  13. Resistance selection, mechanism and stability of Spodoptera litura (Lepidoptera: Noctuidae) to methoxyfenozide.

    PubMed

    Rehan, Adeel; Freed, Shoaib

    2014-03-01

    Methoxyfenozide belongs to a group of biorational insecticides known as insect growth regulators which is used in the control lepidopteran insect pests. Here we report a field collected population of Spodoptera litura selected with methoxyfenozide for thirteen consecutive generations resulted in the development of 83.24 and 2358.6-fold resistance to methoxyfenozide as compared to parental field population and susceptible laboratory population, respectively. The outcomes of synergism studies revealed methoxyfenozide resistance in S. litura to be monooxygenases (MO) mediated with high synergistic ratio (4.83) with piperonyl butoxide (PBO), while S,S,S-tributyl phosphorotrithioate (DEF) showed no synergism with methoxyfenozide (SR=1). This methoxyfenozide resistant strain showed a high cross resistance to deltamethrin (28.82), abamectin (12.87) and little to emamectin benzoate (2.36), however no cross resistance of methoxyfenozide and other tested insecticides was recorded. The results depicted the methoxyfenozide resistance in S. litura to be unstable with high reversion rate which decreased from 2358.6 to 163.9-fold (as compared to susceptible strain) when reared for five generations without any insecticidal exposure. The present research supports the significance of MO-mediated metabolism in resistance to methoxyfenozide, which demands some tactics to tackle this problem. The resistance against methoxyfenozide in S. litura can be overcome by switching off its use for few generations or insecticides rotation having different mode of action. PMID:24759045

  14. Targeting the diuretic hormone receptor to control the cotton leafworm, Spodoptera littoralis.

    PubMed

    Apone, Fabio; Ruggiero, Alessandra; Tortora, Assunta; Tito, Annalisa; Grimaldi, Maria Rosaria; Arciello, Stefania; Andrenacci, Davide; Di Lelio, Ilaria; Colucci, Gabriella

    2014-01-01

    The cotton leafworm, Spodoptera littoralis Boisduval (Lepidoptera: Noctuidae), is one of the most devastating pests of crops worldwide. Several types of treatments have been used against this pest, but many of them failed because of the rapid development of genetic resistance in the different insect populations. G protein coupled receptors have vital functions in most organisms, including insects; thus, they are appealing targets for species-specific pest control strategies. Among the insect G protein coupled receptors, the diuretic hormone receptors have several key roles in development and metabolism, but their importance in vivo and their potential role as targets of novel pest control strategies are largely unexplored. With the goal of using DHR genes as targets to control S. littoralis, we cloned a corticotropin-releasing factor-like binding receptor in this species and expressed the corresponding dsRNA in tobacco plants to knock down the receptor activity in vivo through RNA interference. We also expressed the receptor in mammalian cells to study its signaling pathways. The results indicate that this diuretic hormone receptor gene has vital roles in S. littoralis and represents an excellent molecular target to protect agriculturally-important plants from this pest. PMID:25368043

  15. Global and Local Modulatory Supply to the Mushroom Bodies of the Moth Spodoptera Littoralis

    PubMed Central

    Sinakevitch, Irina; Sjöholm, Marcus; Hansson, Bill S.; Strausfeld, Nicholas J.

    2014-01-01

    The moth Spodoptera littoralis, is a major pest of agriculture whose olfactory system is tuned to odorants emitted by host plants and conspecifics. As in other insects, the paired mushroom bodies are thought to play pivotal roles in behaviors that are elicited by contextual and multisensory signals, amongst which those of specific odors dominate. Compared with species that have elaborate behavioral repertoires, such as the honey bee Apis mellifera or the cockroach Periplaneta americana, the mushroom bodies of S. littoralis were originally viewed as having a simple cellular organization. This has been since challenged by observations of putative transmitters and neuromodulators. As revealed by immunocytology, the spodopteran mushroom bodies like those of other taxa, are subdivided longitudinally into discrete neuropil domains. Such divisions are further supported by the present study, which also demonstrates discrete affinities to different mushroom body neuropils by antibodies raised against two putative transmitters, glutamate and γ-aminobutyric acid, and against three putative neuromodulatory substances: serotonin, A-type allatostatin, and tachykinin-related peptides. The results suggest that in addition to longitudinal divisions of the lobes, circuits in the calyces and lobes are likely to be independently modulated. PMID:18406668

  16. Identification of α-tocopherol and α-tocopheryl acetate from the cuticle of soybean pods armyworm (Spodoptera cosmioides).

    PubMed

    Fronza, Edegar; Migues, Ignacio; Specht, Alexandre; de Barros, Neiva Monteiro; Heinzen, Horacio

    2013-01-01

    The chemical composition of the soybean pods armyworm Spodoptera cosmioides (Walker, 1858) (Lepidoptera: Noctuidae) and Anticarsia gemmatalis Hübner, 1818 (Lepidoptera: Erebidae) larval cuticles was evaluated using gas chromatography coupled to a mass detector (GC-MS). Among the usual lipids found in the insect cuticle, α-tocopherol and α-tocopheryl acetate were also isolated from S. cosmioides. On the other hand, no vitamin E derivative was found in A. gemmatalis exuvia. This is the first report of vitamin E occurrence in the insect's cuticle. PMID:23356865

  17. cDNA cloning and characterization of two trehalases from Spodoptera litura (Lepidoptera; Noctuidade).

    PubMed

    Zou, Q; Wei, P; Xu, Q; Zheng, H Z; Tang, B; Wang, S G

    2013-01-01

    The oriental leafworm moth, Spodoptera litura, is a major agricultural pest in southeast Asia and nearby Pacific regions. Two distinct trehalases have been identified in insects: soluble trehalase (Treh1) and membrane-bound trehalase (Treh2), although there is currently no information on these genes in S. litura. To characterize the distribution and function of treh, cDNAs of Treh proteins were cloned from S. litura. SpoliTreh1 cDNA has an open reading frame of 1758 nucleotides, which encodes a protein of 585 amino acids, with a predicted mass of approximately 67.07 kDa and an isoelectric point of 4.86. SpoliTreh2 cDNA has an open reading frame of 2325 nucleotides, encoding a protein of 645 amino acids, a mass of approximately 73.62 kDa, and an isoelectric point of 5.90. Northern blotting analysis revealed that SpoliTreh1 transcripts are in the midgut, fat body, tracheae, and epidermis, but not in the brain and Malpighian tubules of S. litura larvae, whereas SpoliTreh2 transcripts were found in all 6 tissues. SpoliTreh1 transcripts were highly expressed in the fat body of the pre-pupal stage, and SpoliTreh2 transcripts were highly expressed in the fat body of 3-day-old larvae of the 6th instar and during the 1st 6 days of the pupal stage, except the 2nd day. Both SpoliTreh1 and SpoliTreh2 were highly expressed in third-instar larvae. PMID:23613237

  18. Regulation of Arabidopsis defense responses against Spodoptera littoralis by CPK-mediated calcium signaling

    PubMed Central

    2010-01-01

    Background Plant Ca2+ signals are involved in a wide array of intracellular signaling pathways after pest invasion. Ca2+-binding sensory proteins such as Ca2+-dependent protein kinases (CPKs) have been predicted to mediate the signaling following Ca2+ influx after insect herbivory. However, until now this prediction was not testable. Results To investigate the roles CPKs play in a herbivore response-signaling pathway, we screened the characteristics of Arabidopsis CPK mutants damaged by a feeding generalist herbivore, Spodoptera littoralis. Following insect attack, the cpk3 and cpk13 mutants showed lower transcript levels of plant defensin gene PDF1.2 compared to wild-type plants. The CPK cascade was not directly linked to the herbivory-induced signaling pathways that were mediated by defense-related phytohormones such as jasmonic acid and ethylene. CPK3 was also suggested to be involved in a negative feedback regulation of the cytosolic Ca2+ levels after herbivory and wounding damage. In vitro kinase assays of CPK3 protein with a suite of substrates demonstrated that the protein phosphorylates transcription factors (including ERF1, HsfB2a and CZF1/ZFAR1) in the presence of Ca2+. CPK13 strongly phosphorylated only HsfB2a, irrespective of the presence of Ca2+. Furthermore, in vivo agroinfiltration assays showed that CPK3-or CPK13-derived phosphorylation of a heat shock factor (HsfB2a) promotes PDF1.2 transcriptional activation in the defense response. Conclusions These results reveal the involvement of two Arabidopsis CPKs (CPK3 and CPK13) in the herbivory-induced signaling network via HsfB2a-mediated regulation of the defense-related transcriptional machinery. This cascade is not involved in the phytohormone-related signaling pathways, but rather directly impacts transcription factors for defense responses. PMID:20504319

  19. Establishment and characterization of three embryonic cell lines of beet armyworm, Spodoptera exigua (Lepidoptera: Noctuidae).

    PubMed

    Su, Rui; Zheng, Gui-Ling; Wan, Fang-Hao; Li, Chang-You

    2016-08-01

    Three cell lines (QAU-Se-E-1, -2 and -3, or Se-1, -2 and -3 for short) were established from eggs of beet armyworm (Spodoptera exigua) that have been passaged stably for more than 60 times in TNM-FH medium supplemented with 10 % fetal bovine serum. The cell lines consisted of round and spindle-shaped cells. The round cells accounted for 96.82, 84.34 and 83.16 % of the cells in the three cell lines, respectively, with cell diameters of 16.21 ± 0.72, 15.63 ± 0.58 and 13.06 ± 0.44 μm. Random amplified polymorphic DNA and analysis of the CO I gene showed that the three cell lines were all derived from S. exigua. Growth curves at passage 30 were determined and the results showed that the cell population doubling times were 59.03, 49.08 and 49.91 h, respectively. The three cell lines can be infected by S. exigua multiple nucleopolyhedrovirus (SeMNPV). Se-3 was extremely susceptible to the virus with an infection rate of 97.52 % 4 days after the inoculation and produced 2.02 × 10(6) OBs per mL of culture. Flow cytometry analysis showed that some of Se-1 and Se-2 cells had apoptosis after infection, whereas Se-3 cells did not. Bioassays showed that the virulence of the SeMNPV proliferated from Se-3 was similar to that from the insect with LC50 of 5.55 × 10(5) and 2.64 × 10(5) OBs/mL. Therefore, the cell lines can be used to study the SeMNPV-host interactions and mechanisms underlying the interactions. PMID:25999173

  20. Comparative study of two thioredoxins from common cutworm (Spodoptera litura): cloning, expression, and functional characterization.

    PubMed

    Kang, Tinghao; Wan, Hu; Zhang, Yashu; Shakeel, Muhammad; Lu, Yanhui; You, Hong; Lee, Kwang Sik; Jin, Byung Rae; Li, Jianhong

    2015-04-01

    Thioredoxins (Trxs) are a ubiquitous family of antioxidant enzymes that are involved in protecting organisms against various oxidative stresses. Here, we cloned and characterized two thioredoxins, named SlTrx1 and SlTrx2, from the common cutworm Spodoptera litura. SlTrx1 and SlTrx2, respectively, consist of 988 and 606 bp full-length cDNA with 318 and 447 bp open reading frames encoding 106 and 149 amino acid residues. Furthermore, the N-terminal region of SlTrx2 contains a predicted mitochondrial localization signal (33 amino acids). A phylogenetic relationship analysis revealed that SlTrx1 is in the cytosolic thioredoxin Trx1 cluster, whereas SlTrx2 is in the mitochondrial thioredoxin Trx2 cluster. Recombinant SlTrx1 (14 kDa) and SlTrx2 (16 kDa), expressed in baculovirus-infected insect Sf9 cells, demonstrated insulin disulfide reductase activity at the same optimum temperature and pH value of 35 °C and 7.0, respectively, in vitro. During S. litura development, we found that SlTrx1 and SlTrx2 had similar transcript expression patterns and were constitutively expressed in the epidermis, fat body, and midgut, with the highest expression occurring in the sixth-instar larval stage in the epidermis and midgut. In addition, both SlTrx1 and SlTrx2 mRNA were up-regulated in S. litura after injection with H2O2, cumene hydroperoxide, indoxacarb, and metaflumizone. These results suggest that SlTrx1 and SlTrx2 function as potent antioxidant enzymes, and provide a molecular basis for the roles SlTrx1 and SlTrx2 during development and the oxidative stress response of S. litura. PMID:25542738

  1. Molecular characterization of a ryanodine receptor gene from Spodoptera exigua and its upregulation by chlorantraniliprole.

    PubMed

    Sun, Lina; Qiu, Guisheng; Cui, Li; Ma, Chunsen; Yuan, Huizhu

    2015-09-01

    Chlorantraniliprole is a novel diamide insecticide that targets the insect ryanodine receptor, a Ca(2+) release channel. Spodoptera exigua is a significant insect pest, and chlorantraniliprole is the most popular diamide insecticide used against this pest. To better understand the effects of diamides on RyR expression and [Ca(2+)], we isolated the SeRyR cDNA and investigated changes in SeRyR expression as a result of the application of chlorantraniliprole. The full-length cDNAs of SeRyR contain an open reading frame (ORF) of 15,357 bp with a predicted protein consisting of 5118 amino acids. SeRyR shares 77-92% identity with other insect RyR isoforms and 45-47% identity with vertebrate RyR isoforms. Furthermore, the relative expression abundances of RyR mRNA extracted from S. exigua fat body cells after 24 h of culture in 0.1, 1, 10, 100 nM, 1 µM and 100 µM of chlorantraniliprole changed 1.04-, 0.89-, 1.83-, 2.58-, 4.03- and 3.12-fold compared to blank control, respectively. The regression equation for the relative expression levels of SeRyR after 24 h as a function of the chlorantraniliprole concentration was Y = 0.6455 + 0.8188LgX, R(2) = 0.97093 for the cell line IOZCAS-Spex-II. These results outline the effects of chlorantraniliprole on the expression of SeRyR and provide a basis for the discovery of a compound that may exhibit selective insect activity. PMID:26267053

  2. Analysis of Genes Expression of Spodoptera exigua Larvae upon AcMNPV Infection

    PubMed Central

    Wang, Yong; Zhen, Zou; Tao, Xue Ying; Lee, Joo Hyun; Liu, Qin; Kim, Jae Su; Shin, Sang Woon; Je, Yeon Ho

    2012-01-01

    Background The impact of Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) infection on host gene expression in Spodoptera exigua 4th instar larvae was investigated through the use of 454 sequencing-based RNA-seq of cDNA libraries developed from insects challenged with active AcMNPV or heat-inactivated AcMNPV. Methodology/Principal Findings By comparing the two cDNA libraries, we show that 201 host genes are significantly up-regulated and 234 genes are significantly down-regulated by active AcMNPV infection. Down-regulated host genes included genes encoding antimicrobial peptides, namely three gloverin isoforms and an attacin, indicating that the viral infection actively repressed the expression of a portion of the host immune gene repertoire. Another interesting group of down-regulated host genes included genes encoding two juvenile hormone binding proteins and a hexamerin, all of which are involved in juvenile hormone regulation. The expression of these genes was enhanced by the topical application of Juvenile Hormone III (JHIII) in the insects challenged with heat-inactivated AcMNPV. However, infection with the active virus strongly suppresses the expression of these three genes, regardless of the absence or presence of JHIII. Conclusions/Significance Using RNA-seq, we have identified groups of immune-regulated and juvenile hormone-regulated genes that are suppressed by infection with active AcMNPV. This information and further studies on the regulation of host gene expression by AcMNPV will provide the tools needed to enhance the utility of the virus as an effective protein expression system and as an insecticide. PMID:22860129

  3. Identification and functional characterization of sex pheromone receptors in beet armyworm Spodoptera exigua (Hübner).

    PubMed

    Liu, Chengcheng; Liu, Yang; Walker, William B; Dong, Shuanglin; Wang, Guirong

    2013-08-01

    In moths, males can detect a distinct blend of several pheromone components by specialized olfactory receptor neurons (ORNs) on the antennae. Four candidate pheromone receptors (PR) with seven transmembrane domains were identified by homology cloning from the antennae of Spodoptera exigua (Sexi). Phylogenetic analyses reveal that all four odorant receptors (OR) belong to pheromone receptor subtypes. Expression patterns revealed that PRs were male-specific in the antenna except for SexiOR11, which was female antenna-biased. Functional analyses of these PRs were conducted using heterologous expression in Xenopus oocytes. SexiOR13 and SexiOR16 were all broadly activated by multiple pheromone components. SexiOR13 responded robustly to the critical pheromone component, Z9, E12-14:OAc and the minor pheromone component, Z9-14:OAc at a concentration of 10(-4) M. Dose-response studies indicate that SexiOR13 was approximately 4 times more sensitive to Z9,E12-14:OAc (EC50 = 3.158 × 10(-6) M) compared to Z9-14:OAc (EC50 = 1.203 × 10(-5) M). While, SexiOR16 responded robustly to the secondary pheromone component Z9-14:OH with high sensitivity (EC50 = 9.690 × 10(-7) M). However, similar tests of the five pheromones with SexiOR6 and SexiOR11 failed to elicit any response. These results provide basic knowledge to further advance research on the molecular mechanisms of pheromone reception. PMID:23751753

  4. In Vivo Pyro-SIP Assessing Active Gut Microbiota of the Cotton Leafworm, Spodoptera littoralis

    PubMed Central

    Shao, Yongqi; Arias-Cordero, Erika; Guo, Huijuan; Bartram, Stefan; Boland, Wilhelm

    2014-01-01

    The gut microbiota is of crucial importance for the host with considerable metabolic activity. Although great efforts have been made toward characterizing microbial diversity, measuring components' metabolic activity surprisingly hasn't kept pace. Here we combined pyrosequencing of amplified 16S rRNA genes with in vivo stable isotope probing (Pyro-SIP) to unmask metabolically active bacteria in the gut of cotton leafworm (Spodoptera littoralis), a polyphagous insect herbivore that consumes large amounts of plant material in a short time, liberating abundant glucose in the alimentary canal as a most important carbon and energy source for both host and active gut bacteria. With 13C glucose as the trophic link, Pyro-SIP revealed that a relatively simple but distinctive gut microbiota co-developed with the host, both metabolic activity and composition shifting throughout larval stages. Pantoea, Citrobacter and Clostridium were particularly active in early-instar, likely the core functional populations linked to nutritional upgrading. Enterococcus was the single predominant genus in the community, and it was essentially stable and metabolically active in the larval lifespan. Based on that Enterococci formed biofilm-like layers on the gut epithelium and that the isolated strains showed antimicrobial properties, Enterococcus may be able to establish a colonization resistance effect in the gut against potentially harmful microbes from outside. Not only does this establish the first in-depth inventory of the gut microbiota of a model organism from the mostly phytophagous Lepidoptera, but this pilot study shows that Pyro-SIP can rapidly gain insight into the gut microbiota's metabolic activity with high resolution and high precision. PMID:24475063

  5. Effects of green manure cover crops on Spodoptera litura (Lepidoptera: Noctuidae) populations.

    PubMed

    Tuan, Shu-Jen; Li, Nian-Jhen; Yeh, Chih-Chun; Tang, Li-Cheng; Chi, Hsin

    2014-06-01

    Spodoptera litura (F.) is an important pest of numerous agro-economic crops, including green manure cover crops. In Taiwan, sesbania (Sesbanin roxburghii Merr.), sunn hemp (Crotalaria juncea L.), and rapeseed (Brassicae campestris L. variety chinensis) are the most popular green manure crops; sesbania and sunn hemp are commonly planted in warm seasons, whereas rapeseed is grown in the winter. In this study, life-table data for S. litura reared on these three green manures were collected to evaluate their roles as refuges of this pest. The net reproductive rate, intrinsic rate of increase, and finite rate of increase of S. litura were the highest when reared on sesbania (1428.1 offspring, 0.2327 d(-1), 1.2621 d(-1)), followed by sunn hemp (778.4 offspring, 0.2070 d(-1), 1.2300 d(-1)) and rapeseed (737.6 offspring, 0.2040 d(-1), 1.2263 d(-1)). The high growth rates on these green manure crops show that they can serve as potential breeding sites for S. litura. Population projection demonstrated the rapid growth of S. litura on sesbania, sunn hemp, and rapeseed as well. Because most growers have traditionally ignored pest management in green manure fields, the mass emergence of S. litura in these fields may cause unexpected infestations in nearby vegetable, corn, and peanut crops. This study shows that the use of green manures as sources of nutrients should be critically reassessed and an area-wide pest management program should be instituted by taking the population of S. litura in green manure fields into consideration. PMID:25026645

  6. Demography and Consumption of Spodoptera litura (Lepidoptera: Noctuidae) Reared on Cabbage and Taro.

    PubMed

    Tuan, Shu-Jen; Yeh, Chih-Chun; Atlihan, Remzi; Chi, Hsin; Tang, Li-Cheng

    2016-04-01

    Spodoptera litura (F.) causes considerable economic damage to multiple agro-crops annually in many countries. In this study, the demography of S. litura reared on cabbage and taro was investigated using the age-stage, two-sex life table at 25±1°C, 60±10% relative humidity, and a photoperiod of 12: 12 (L:D) h. Our results showed that the net reproductive rate, intrinsic rate, and finite rate of population increase on cabbage (1893.1 offspring, 0.2374 d(−1), and 1.2679 d(−1)) were all not significantly different from those on taro (1361.0 offspring, 0.2298 d(−1), and 1.2584 d(−1)). The net consumption rate on cabbage (439.1 cm2) was, however, three times higher than that on taro (141.7 cm2). According to the population parameters, both cabbage and taro are suitable host plants for S. litura. When both the population growth rate and the consumption rate were taken into consideration, the finite consumption rate on cabbage (ω=3.8054) was significantly higher than that on taro (ω=1.3184). In Taiwan, taro and cabbage are commonly planted in adjacent farm plots, with taro being grown from March to November and cabbage from October to April. Because of the overlapping growth periods of the two crops, S. litura can easily propagate throughout the year by switching between the adjacent crops during the overlap periods. Pest management strategies for controlling S. litura must be thoroughly reevaluated based on ecological characteristics, including its life table and consumption rate on its major host plants. PMID:26578628

  7. Growth Performance and Biometric Characteristics of Spodoptera litura (Lepidoptera: Noctuidae) Reared on Different Host Plants.

    PubMed

    Tuan, Shu-Jen; Li, Nian-Jhen; Yeh, Chih-Chun

    2015-10-01

    Spodoptera litura (F.), an important polyphagous insect pest, attacks ca. 300 economic crops in dozens of countries. Investigations into its growth and development performance on different host plants can provide an understanding of the potential for increase of S. litura population in the field. We measured the development time, body weight, and head capsule width of S. litura larvae reared on cabbage, taro, peanut, and sesbania, a green manure. Larvae reared on cabbage ingested a significantly higher amount of protein and completed the immature stages in a shorter period than those reared on the other three plants. The relationship between head capsule width and larval instars on these four crops fitted well with Dyar's rule, and the Dyar's ratios ranged from 1.4554 to 1.6786, although a few supernumerary instar individuals on sesbania, peanut, and taro showed lower ratios (1.0103 to 1.1330). The head capsule width among cohorts fed on different host plants varied significantly and overlapped between late instars, which could lead to a misjudgment of instar stage in the field. The growth index of S. litura on cabbage was significantly higher than on the other host plants. Larvae fed on sesbania showed the highest feeding index and a better growth index than on taro and peanut. We therefore suggest that the area-wide pest management against S. litura should take into consideration its occurrence on sesbania in intercropping seasons. Additionally, the effective management of this pest during cropping windows between all these four plants should not be ignored. PMID:26453712

  8. Response of the Cutworm Spodoptera litura to Sesame Leaves or Crude Extracts in Diet

    PubMed Central

    Ofosuhene Sintim, Henry; Tashiro, Toru; Motoyama, Naoki

    2009-01-01

    The effects of extracts of sesame, Sesamum indicum L. (Liamiales: Pedaliaceae), and whole leaves of some selected cultivars of sesame were tested using a natural host Spodoptera litura (F.) (Lepidoptera: Noctuidae). Indices taken using the immature stages include; diet utilization, growth and development and induction of detoxification enzymes. The results indicate that S. litura generally selects its food amongst cultivars within 6 hours after food presentation. Growth and development of the insect is controlled also by plant acceptability and quality. Although all the cultivars tested significantly limit insect growth and development the variety 56S-radiatum did not allow a complete life cycle as pupation from first instar stage was 0%. Generally the crucial period for immature S. litura was the larval period, especially the first two instars where the weight of an insect fed on an experimental diet was three times lower than that of a control diet. The larval developmental period was greater than 40 days as compared to 17 days for insects fed a control diet. S. litura also had lowered efficiency in utilizing ingested food, from a low of 13% in a sesame cultivar to 45% in the control diet. The key detoxification enzyme was a glutathione s-transferase that was confirmed by a 6-fold increase between S. litura fed a plant cultivar vs. a control diet towards the substrate 1,2-dichloro-4-nitrobenzene. First and second instars of S. litura have a relatively reduced detoxification of enzymes in response to plant cultivar diets leading to low survival. A 3% v/w crude extract of the cultivars increased enzyme induction towards all the tested substrates. PMID:20050772

  9. Constitutive activation of the midgut response to Bacillus thuringiensis in Bt-resistant Spodoptera exigua.

    PubMed

    Hernández-Martínez, Patricia; Navarro-Cerrillo, Gloria; Caccia, Silvia; de Maagd, Ruud A; Moar, William J; Ferré, Juan; Escriche, Baltasar; Herrero, Salvador

    2010-01-01

    Bacillus thuringiensis is the most effective microbial control agent for controlling numerous species from different insect orders. The main threat for the long term use of B. thuringiensis in pest control is the ability of insects to develop resistance. Thus, the identification of insect genes involved in conferring resistance is of paramount importance. A colony of Spodoptera exigua (Lepidoptera: Noctuidae) was selected for 15 years in the laboratory for resistance to Xentari™, a B. thuringiensis-based insecticide, reaching a final resistance level of greater than 1,000-fold. Around 600 midgut ESTs were analyzed by DNA-macroarray in order to find differences in midgut gene expression between susceptible and resistant insects. Among the differentially expressed genes, repat and arylphorin were identified and their increased expression was correlated with B. thuringiensis resistance. We also found overlap among genes that were constitutively over-expressed in resistant insects with genes that were up-regulated in susceptible insects after exposure to Xentari™, suggesting a permanent activation of the response to Xentari™ in resistant insects. Increased aminopeptidase activity in the lumen of resistant insects in the absence of exposure to Xentari™ corroborated the hypothesis of permanent activation of response genes. Increase in midgut proliferation has been proposed as a mechanism of response to pathogens in the adult from several insect species. Analysis of S. exigua larvae revealed that midgut proliferation was neither increased in resistant insects nor induced by exposure of susceptible larvae to Xentari™, suggesting that mechanisms other than midgut proliferation are involved in the response to B. thuringiensis by S. exigua larvae. PMID:20862260

  10. Constitutive Activation of the Midgut Response to Bacillus thuringiensis in Bt-Resistant Spodoptera exigua

    PubMed Central

    Hernández-Martínez, Patricia; Navarro-Cerrillo, Gloria; Caccia, Silvia; de Maagd, Ruud A.; Moar, William J.; Ferré, Juan; Escriche, Baltasar; Herrero, Salvador

    2010-01-01

    Bacillus thuringiensis is the most effective microbial control agent for controlling numerous species from different insect orders. The main threat for the long term use of B. thuringiensis in pest control is the ability of insects to develop resistance. Thus, the identification of insect genes involved in conferring resistance is of paramount importance. A colony of Spodoptera exigua (Lepidoptera: Noctuidae) was selected for 15 years in the laboratory for resistance to Xentari™, a B. thuringiensis-based insecticide, reaching a final resistance level of greater than 1,000-fold. Around 600 midgut ESTs were analyzed by DNA-macroarray in order to find differences in midgut gene expression between susceptible and resistant insects. Among the differentially expressed genes, repat and arylphorin were identified and their increased expression was correlated with B. thuringiensis resistance. We also found overlap among genes that were constitutively over-expressed in resistant insects with genes that were up-regulated in susceptible insects after exposure to Xentari™, suggesting a permanent activation of the response to Xentari™ in resistant insects. Increased aminopeptidase activity in the lumen of resistant insects in the absence of exposure to Xentari™ corroborated the hypothesis of permanent activation of response genes. Increase in midgut proliferation has been proposed as a mechanism of response to pathogens in the adult from several insect species. Analysis of S. exigua larvae revealed that midgut proliferation was neither increased in resistant insects nor induced by exposure of susceptible larvae to Xentari™, suggesting that mechanisms other than midgut proliferation are involved in the response to B. thuringiensis by S. exigua larvae. PMID:20862260

  11. An antenna-biased carboxylesterase is specifically active to plant volatiles in Spodoptera exigua.

    PubMed

    He, Peng; Zhang, Ya-Nan; Yang, Ke; Li, Zhao-Qun; Dong, Shuang-Lin

    2015-09-01

    Odorant-degrading enzymes (ODEs) in sensillar lymph are proposed to play important roles in the maintenance of the sensitivity of the olfactory sensilla, by timely degrading the odorants that have already fulfilled the activation of the odorant receptor (OR). Here we reported the cloning and characterization of an ODE gene (SexiCXE10) from the polyphagous insect pest Spodoptera exigua. SexiCXE10 is a carboxylesterase (CXE) gene, encoding a protein with 538 amino acid residues, and bearing typical characteristics of Carboxyl/cholinesterase (CCE, EC 3.1.1.1.) gene family. Tissue-temporal expression pattern by qPCR revealed that the SexiCXE10 mRNA was highly antenna biased, and maintained at high level throughout the adult stage. Further fluorescence in situ hybridization demonstrated that SexiCXE10 mRNA signal was detected under sensilla basiconica and short and long sensilla trichodea. Finally, enzymatic study using purified recombinant enzyme showed that SexiCXE10 had high activity specifically for ester plant volatiles with 7-10 carbon atoms, while no activity was found with S. exigua sex pheromone components and plant volatiles with more carbon atoms. In addition, SexiCXE10 displayed lower activity at acidic pH (pH 5.0), while higher activity was found at neutral and alkaline conditions (pH 6.5-9.0). Our results suggest that SexiCXE10 may play an important role in the degradation of the host plant volatiles, and thus contributes to the high sensitivity of the olfactory system in S. exigua. Meanwhile, the CXE would be a potential target for developing behavioral antagonists and pesticides against S. exigua. PMID:26267057

  12. Identification of Spodoptera exigua nucleopolyhedrovirus genes involved in pathogenicity and virulence.

    PubMed

    Serrano, Amaya; Pijlman, Gorben P; Vlak, Just M; Muñoz, Delia; Williams, Trevor; Caballero, Primitivo

    2015-03-01

    Genome sequence analysis of seven different Spodoptera exigua multiple nucleopolyhedrovirus (SeMNPV) isolates that differed in insecticidal phenotype permitted the identification of genes likely to be involved in pathogenicity of occlusion bodies (OBs) and speed of kill (virulence) of this virus: se4 (hoar), se5 (unknown function), se28 (unknown function), se76 (cg30), se87 (p26) and se129 (p26). To study the role of these genes experimentally on the insecticidal phenotype, a bacmid-based recombination system was constructed to delete selected genes from a SeMNPV isolate, VT-SeAL1, designated as SeBacAL1. All of the knockout viruses were viable and the repair viruses behaved like the wild-type control, vSeBacAL1. Deletion of se4, se5, se76 and se129 resulted in decreased OB pathogenicity compared to vSeBacAL1 OBs. In contrast, deletion of se87 did not significantly affect OB pathogenicity, whereas deletion of se28 resulted in significantly increased OB pathogenicity. Deletion of se4, se28, se76, se87 and se129 did not affect speed of kill compared to the bacmid vSeBacAL1, whereas speed of kill was significantly extended following deletion of se5 and in the wild-type isolate (SeAL1), compared to that of the bacmid. Therefore, biological assays confirmed that several genes had effects on virus insecticidal phenotype. Se5 is an attractive candidate gene for further studies, as it affects both biological parameters of this important biocontrol virus. PMID:25644432

  13. Oviposition by Spodoptera exigua on Nicotiana attenuata primes induced plant defence against larval herbivory.

    PubMed

    Bandoly, Michele; Hilker, Monika; Steppuhn, Anke

    2015-08-01

    Plants exhibit multifarious defence traits against herbivory that are constitutively expressed or induced upon attack. Insect egg deposition often precedes impending larval attack, and several plants can increase their resistance against larvae after experiencing the oviposition by an herbivore. The nature of such oviposition-mediated resistance remains unknown, and here we aim to determine plant traits that explain it. We test whether oviposition on a host plant can induce plant defence responses or enhance (prime) the induction of defence traits in response to larval herbivory. We exposed Nicotiana attenuata plants to oviposition by moths of a generalist herbivore, Spodoptera exigua. Its larvae suffered higher mortality, retarded development and inflicted less feeding damage on oviposition-experienced than on oviposition-unexperienced plants. While oviposition alone did not induce any of the examined defence traits, oviposited plants exhibited a stronger inducibility of known defence traits, i.e. caffeoylputrescine (CP) and trypsin protease inhibitors (TPIs). We found no effects of oviposition on phytohormone levels, but on the feeding-inducible accumulation of the transcription factor NaMyb8 that is governing biosynthesis of phenylpropanoid-polyamine conjugates, including CP. Comparison of larval performance on wild-type plants, CP-deficient plants (silenced NaMyb8 gene), and TPI-deficient plants (silenced NaPI gene) revealed that priming of plant resistance to larvae by prior oviposition required NaMyb8-mediated defence traits. Our results show that plants can use insect egg deposition as a warning signal to prime their feeding-induced defence. PMID:26096574

  14. Differential immunosuppression by Campoletis chlorideae eggs and ichnovirus in larvae of Helicoverpa armigera and Spodoptera exigua.

    PubMed

    Han, Li-Bin; Yin, Li-Hong; Huang, Ling-Qiao; Wang, Chen-Zhu

    2015-09-01

    The ichneumonid wasp, Campoletis chlorideae Uchida, successfully develops in the cotton bollworm Helicoverpa armigera (Hübner), but rarely survives in the beet armyworm Spodoptera exigua (Hübner) due to the encapsulation by host immunity. In this study, we investigated the role of C. chlorideae ichnovirus (CcIV) and eggs in the evasion of the host immune system. Washed eggs of different types, immature, mature, newly laid, or pretreated with protease K, were injected alone or with the calyx fluid containing CcIV into the larvae of H. armigera and S. exigua. In H. armigera, when injected with washed eggs alone, only 9.5% of the mature eggs were encapsulated at 24h post-injection. This is much lower than that of the immature eggs (100%), mature eggs pretreated with protease K (100%) and newly laid eggs (54.4%). No encapsulation was observed when the washed eggs were co-injected with calyx fluid at 24h post-injection. Conversely, the eggs in all treatments were encapsulated in S. exigua. Electron microscopic observations of parasitoid eggs showed structural differences between the surfaces of the mature and other kinds of eggs. The injected CcIV decreased the numbers of host hemocytes and suppressed the spreading ability of plasmatocytes and granulocytes in H. armigera, but had little effect on the hemocytes from S. exigua. In conclusion, the C. chlorideae egg provides a passive protection against encapsulation by itself, and CcIV supplies an active protection through disrupting host immune responses. These coordinated protections are host-specific, implying their role in host range determination. PMID:26183795

  15. Cross-resistance and Inheritance of Resistance to Emamectin Benzoate in Spodoptera exigua (Lepidoptera: Noctuidae).

    PubMed

    Che, Wunan; Huang, Jianlei; Guan, Fang; Wu, Yidong; Yang, Yihua

    2015-08-01

    Beet armyworm, Spodoptera exigua (Hübner), is a worldwide pest of many crops. Chemical insecticides are heavily used for its control in China, and serious resistance has been evolved in the field to a variety of insecticides including emamectin benzoate. Through repeated backcrossing to a susceptible strain (WH-S) and selection with emamectin benzoate, the trait conferring resistance to emamectin benzoate in a field-collected population of S. exigua (moderately resistant to emamectin benzoate and strongly resistant to pyrethroids and indoxacarb) was introgressed into WH-S to generate a near-isogenic resistant strain (WH-EB). Compared with WH-S, the WH-EB strain developed a 1,110-fold resistance to emamectin benzoate and a high level of cross-resistance to abamectin (202-fold), with low levels of cross-resistance to cypermethrin (10-fold) and chlorfluazuron (7-fold), but no cross-resistance to representatives of another six different classes of insecticides (chlorantraniliprole, chlorfenapyr, indoxacarb, spinosad, tebufenozide, and chlorpyrifos). Resistance to emamectin benzoate in WH-EB was autosomal, incompletely dominant, and polygenic. Limited cross-resistance in WH-EB indicates that emamectin benzoate can be rotated with other classes of insecticides to which it does not show cross-resistance to delay the evolution of resistance in S. exigua. The incompletely dominant nature of resistance in S. exigua may explain the rapid evolution of resistance to emamectin benzoate in the field, and careful deployment of this chemical within a resistance management program should be considered. PMID:26470348

  16. Testing the joint effects hypothesis of elemental defense using Spodoptera exigua.

    PubMed

    Cheruiyot, Dorothy J; Boyd, Robert S; Moar, William

    2015-02-01

    Metal hyperaccumulation may be an elemental defense, in which high concentrations of a metal in plant tissues decrease herbivore survival or growth rate. The Joint Effects Hypothesis suggests that a combination of metals, or a combination of a metal with an organic compound, may have an enhanced defensive effect. The enhancement may be additive or synergistic: in either case the concentration of a particular metal necessary to provide a defensive benefit for the plant is lowered. We tested the Joint Effects Hypothesis using Spodoptera exigua (beet armyworm) neonates fed artificial diets. Metal + metal experiments utilized diets amended with metal pairs, using four metals commonly hyperaccumulated by plants (Co, Cu, Ni, and Zn). We also conducted metal + organic compound experiments, pairing each metal with nicotine, mustard seed powder, or tannic acid. We tested for joint effects using both lethal (LC20 levels) and sublethal concentrations (10-25 % reduced larval weight) of the chemicals tested. For all experiments, either additive or synergistic effects were found. Of the metal + metal pairs tested, three (Co + Cu, Cu + Zn, and Ni + Zn) were synergistic in lethal concentration tests and only Co + Cu was synergistic in sublethal tests. For metal + organic combination lethal tests, synergism occurred for all combinations except for Co or Ni + nicotine, Ni + mustard seed powder, and Zn + nicotine. For sublethal tests, Zn + all three organic chemicals, Co + mustard seed powder or tannic acid, and Cu + nicotine, were synergistic. These results support the Joint Effects Hypothesis, suggesting that metals combined with other metals or organic compounds may be more effective against herbivores than individual metals. PMID:25712748

  17. Identification and Characterization of Candidate Chemosensory Gene Families from Spodoptera exigua Developmental Transcriptomes.

    PubMed

    Liu, Nai-Yong; Zhang, Ting; Ye, Zhan-Feng; Li, Fei; Dong, Shuang-Lin

    2015-01-01

    Insect chemosensory genes have been considered as potential molecular targets to develop alternative strategies for pest control. However, in Spodoptera exigua, a seriously polyphagous agricultural pest, only a small part of such genes have been identified and characterized to date. Here, using a bioinformatics screen a total of 79 chemosensory genes were identified from a public transcriptomic data of different developmental stages (eggs, 1st to 5th instar larvae, pupae, female and male adults), including 34 odorant binding proteins (OBPs), 20 chemosensory proteins (CSPs), 22 chemosensory receptors (10 odorant receptors (ORs), six gustatory receptors (GRs) and six ionotropic receptors (IRs)) and three sensory neuron membrane proteins (SNMPs). Notably, a new group of lepidopteran SNMPs (SNMP3 group) was found for the first time in S. exigua, and confirmed in four other moth species. Further, reverse transcription PCR (RT-PCR) and quantitative real time PCR (qPCR) were employed respectively to validate the sequences and determine the expression patterns of 69 identified chemosensory genes regarding to sexes, tissues and stages. Results showed that 67 of these genes could be detected and reconstructed in at least one tissue tested. Further, 60 chemosensory genes were expressed in adult antennae and 52 in larval heads with the antennae, whereas over half of the genes were also detected in non-olfactory tissues like egg and thorax. Particularly, S. exigua OBP2 showed a predominantly larval head-biased expression, and functional studies further indicated its potentially olfactory roles in guiding food searching of larvae. This work suggests functional diversities of S. exigua chemosensory genes and could greatly facilitate the understanding of olfactory system in S. exigua and other lepidopteran species. PMID:26221071

  18. Herbivore-induced maize leaf volatiles affect attraction and feeding behavior of Spodoptera littoralis caterpillars.

    PubMed

    von Mérey, Georg E; Veyrat, Nathalie; D'Alessandro, Marco; Turlings, Ted C J

    2013-01-01

    Plants under herbivore attack emit volatile organic compounds (VOCs) that can serve as foraging cues for natural enemies. Adult females of Lepidoptera, when foraging for host plants to deposit eggs, are commonly repelled by herbivore-induced VOCs, probably to avoid competition and natural enemies. Their larval stages, on the other hand, have been shown to be attracted to inducible VOCs. We speculate that this contradicting behavior of lepidopteran larvae is due to a need to quickly find a new suitable host plant if they have fallen to the ground. However, once they are on a plant they might avoid the sites with fresh damage to limit competition and risk of cannibalism by conspecifics, as well as exposure to natural enemies. To test this we studied the effect of herbivore-induced VOCs on the attraction of larvae of the moth Spodoptera littoralis and on their feeding behavior. The experiments further considered the importance of previous feeding experience on the responses of the larvae. It was confirmed that herbivore-induced VOCs emitted by maize plants are attractive to the larvae, but exposure to the volatiles decreased the growth rate of caterpillars at early developmental stages. Larvae that had fed on maize previously were more attracted by VOCs of induced maize than larvae that had fed on artificial diet. At relatively high concentrations synthetic green leaf volatiles, indicative of fresh damage, also negatively affected the growth rate of caterpillars, but not at low concentrations. In all cases, feeding by the later stages of the larvae was not affected by the VOCs. The results are discussed in the context of larval foraging behavior under natural conditions, where there may be a trade-off between using available host plant signals and avoiding competitors and natural enemies. PMID:23825475

  19. Effects of Pyriproxyfen on Female Reproduction in the Common Cutworm, Spodoptera litura (F.) (Lepidoptera: Noctuidae)

    PubMed Central

    Xu, Qi; Tang, Bin; Zou, Qi; Zheng, Huizhen; Liu, Xiaojun; Wang, Shigui

    2015-01-01

    The common cutworm, Spodoptera litura, is a rapidly reproducing pest of numerous agricultural ecosystems worldwide. The use of pesticides remains the primary means for controlling S. litura, despite their negative ecological impact and potential threat to human health. The use of exogenous hormone analogs may represent an alternative to insecticides. Juvenile hormones (JHs) play an important role in the reproductive systems of female insects, but the effects of pyriproxyfen, a JH analog, on reproduction in S. litura were poorly understood. In this paper, we topically treated the newly emerged females with 20, 60, or 100 μg of pyriproxyfen to determine its effects on reproduction. Then, we examined the expression of vitellogenin (Vg) and three hormone receptors, USP, HR3, and EcR, using quantitative reverse transcription and real-time polymerase chain reaction (qRT-PCR), and found that pyriproxyfen up-regulated the expression of Vg, USP, and HR3, whereas the expression of EcR was unaffected. An analysis of fecundity showed that the peak oviposition day, lifespan, and oviposition period were progressively shortened as the pyriproxyfen dosage increased. We also found that pyriproxyfen decreased egg laying amount, whereas the number of mature eggs that remained in the ovarioles of dead females increased as the pyriproxyfen dosage increased. We examined oocytes using transmission electron microscopy and found that treatment with 100 μg of pyriproxyfen increased the metabolism by increasing the amount of rough endoplasmic reticulum and mitochondria in the primary oocytes. Our results suggest that the topical application of pyriproxyfen on newly emerged females can efficiently reduce reproduction in S. litura and may represent an alternative to the use of insecticides for controlling the agricultural pest. PMID:26444432

  20. Various eicosanoids modulate the cellular and humoral immune responses of the beet armyworm, Spodoptera exigua.

    PubMed

    Shrestha, Sony; Kim, Yonggyun

    2009-09-01

    Cyclooxygenase (COX) and lipoxygenase (LOX) can catalyze the oxidation of C20 fatty acids to produce certain eicosanoids, which play roles in mediating immune responses in insects. Despite their critical role in insect immunity, there have been few studies of the unique effects of different eicosanoids on immune responses. This study analyzed cellular and humoral immune responses of the beet armyworm, Spodoptera exigua, using seven eicosanoids selected from two major eicosanoid subgroups: prostaglandin (PG) and leukotriene (LT), derived from catalytic activities of COX and LOX respectively. Upon bacterial challenge, all seven eicosanoids (PGA(1), PGB(2), PGD(2), PGE(1), PGE(2), PGF(1alpha), and LTB(4)) significantly induced hemocyte nodulation and phagocytosis in the presence of dexamethasone, an eicosanoid biosynthesis inhibitor. However, only PGs induced cell lysis of oenocytoids to release prophenoloxidase, which resulted in an increase in phenoloxidase activity. These seven eicosanoids also induced expression of humoral immune-associated genes, including prophenoloxidase, serpin, dopa decarboxylase, cecropin, and lysozyme, in which PGB(2) and PGE(1) did not induce gene expression of prophenoloxidase. To understand the interactions between different eicosanoids, mixture effects of these eicosanoids were compared with their individual eicosanoid effects on mediating nodule formation in response to bacterial challenge. All six single PGs showed increases in nodule formation in a dose-dependent manner without significant difference among the different types. LTB(4) was more potent than the tested PGs in mediating the cellular immune response. At low doses, all combinations of two eicosanoids showed significant additive effects on nodule formation. These results indicate that immune target cells, such as hemocyte and fat body, of S. exigua can respond to different COX and LOX products to express cellular and humoral immune responses, and their overlapping, additive

  1. Toxicity of Alangium salvifolium Wang chemical constituents against the tobacco cutworm Spodoptera litura Fab.

    PubMed

    Selin-Rani, Selvaraj; Senthil-Nathan, Sengottayan; Revathi, Kannan; Chandrasekaran, Rajamanickam; Thanigaivel, Annamalai; Vasantha-Srinivasan, Prabhakaran; Ponsankar, Athirstam; Edwin, Edward Sam; Pradeepa, Venkatraman

    2016-01-01

    Widespread use of synthetic pesticides has resulted in the development of insecticide-resistant populations of pests and harmful effects on human health and the environment. There is a need to identify alternative pest management strategies to reduce our reliance on conventional chemical pesticides. In recent years the use of botanical pesticides for protecting crops from insect pests has assumed greater importance. Methanol extract of Alangium salvifolium (L.f.) Wang has potential insecticidal activity against Spodoptera litura Fab. The active fractions were identified through chromatographic techniques as F-IV (Rf value=0.45) and F-VI (Rf value=0.63) and were subjected to GC-MS (GCMATE II). Fifty, 100 and 200ppm of active fractions were applied to fourth instar larvae and the mortality increased with higher concentrations. Relative consumption rate, relative growth rate, efficiency of conversion of ingested food and efficiency of conversion of digested food values all decreased in treated larvae, but approximate digestibility rate increased after treatment. The hydrolytic enzymes, such as acid phosphatase, alkaline phosphatase and the glycolytic enzyme lactate dehydrogenase were inhibited in treated larvae compared with controls. The histopathology study revealed that the epithelial columnar cells were enlarged, completely atrophied; intercellular spaces were swollen, and also noted a cytoplasmic ooze of cell material that mixed with food column. The present study clearly showed the active fractions from A. salvifolium as potential botanicals to control the larvae of S. litura. This is the first report for nutritional indices, enzymatic activities and histological effects of A. salvifolium chemical constituents against S. litura. Thus probably, this will be used as an alternative for synthetic pesticides against the polyphagous pest like S. litura. PMID:26778440

  2. Peroxiredoxin 5 from common cutworm (Spodoptera litura) acts as a potent antioxidant enzyme.

    PubMed

    Wan, Hu; Kang, Tinghao; Zhan, Sha; You, Hong; Zhu, Fuxing; Lee, Kwang Sik; Zhao, Haigang; Jin, Byung Rae; Li, Jianhong

    2014-09-01

    In this study, we describe the cloning and characterization of a Prx from the common cutworm Spodoptera litura (SlPrx5). The SlPrx5 cDNA contains an open reading frame of 477 bp encoding a predicted protein of 159 amino acid residues, 16.902 kDa, and an isoelectric point of 7.68. Furthermore, the deduced amino acid sequence of the SlPrx5 cDNA showed 86% identity to Papilio xuthus Prx5, 72% to Aedes aegypti Prx5, and 64-67% to other insect Prxs. A phylogenetic analysis further revealed that the deduced amino acid sequence of SlPrx5 groups within the atypical 2-Cys Prx cluster. Recombinant SlPrx5 (20 kDa) purified from baculovirus-infected insect cells was found to reduce H2O2 in the presence of electrons donated by dithiothreitol and protect super-coiled DNA from damage by metal-catalyzed oxidation in vitro. During S. litura development, SlPrx5 is constitutively expressed in the epidermis, fat body, and midgut, with the highest expression occurring in the sixth-instar larval stage in the fat body and midgut. Additionally, SlPrx5 mRNA expression was up-regulated after injection with H2O2, cumene hydroperoxide, indoxacarb, and metaflumizone. A disc diffusion assay indicated that recombinant SlPrx5 can play a functional role in protecting cells from oxidative stress in vivo. These results provide insight into the role of SlPrx5 during development and the oxidative stress response of S. litura. PMID:24998343

  3. Identification and Differential Expression of a Candidate Sex Pheromone Receptor in Natural Populations of Spodoptera litura

    PubMed Central

    Lin, Xinda; Zhang, Qinhui; Wu, Zhongnan; Du, Yongjun

    2015-01-01

    Olfaction is primarily mediated by highly specific olfactory receptors (ORs), a subfamily of which are the pheromone receptors that play a key role in sexual communication and can contribute to reproductive isolation. Here we cloned and identified an olfactory receptor, SlituOR3 (Genbank NO. JN835270), from Spodoptera litura, to be the candidate pheromone receptor. It exhibited male-biased expression in the antennae, where they were localized at the base of sensilla trichoidea. Conserved orthologues of these receptors were found amongst known pheromone receptors within the Lepidoptera, and SlituOR3 were placed amongst a clade of candidate pheromone receptors in a phylogeny tree of insect ORs. SlituOR3 is required for the EAG responses to both Z9E11-14:OAc and Z9E12-14:OAc SlituOR3 showed differential expression in S. litura populations attracted to traps baited with a series of sex pheromone blends composed of different ratios of (9Z,11E)-tetradecadienyl acetate (Z9E11-14:OAc) and (9Z,12E)-tetradecadienyl acetate (Z9E12-14:OAc). The changes in the expression level of SlitOR3 and antennal responses after SlitOR3 silencing suggested that SlitOR3 is required for the sex pheromone signaling. We infer that variation in transcription levels of olfactory receptors may modulate sex pheromone perception in male moths and could affect both of pest control and monitoring efficiency by pheromone application after long time mass trapping with one particular ratio of blend in the field. PMID:26126192

  4. Identification and Characterization of Candidate Chemosensory Gene Families from Spodoptera exigua Developmental Transcriptomes

    PubMed Central

    Liu, Nai-Yong; Zhang, Ting; Ye, Zhan-Feng; Li, Fei; Dong, Shuang-Lin

    2015-01-01

    Insect chemosensory genes have been considered as potential molecular targets to develop alternative strategies for pest control. However, in Spodoptera exigua, a seriously polyphagous agricultural pest, only a small part of such genes have been identified and characterized to date. Here, using a bioinformatics screen a total of 79 chemosensory genes were identified from a public transcriptomic data of different developmental stages (eggs, 1st to 5th instar larvae, pupae, female and male adults), including 34 odorant binding proteins (OBPs), 20 chemosensory proteins (CSPs), 22 chemosensory receptors (10 odorant receptors (ORs), six gustatory receptors (GRs) and six ionotropic receptors (IRs)) and three sensory neuron membrane proteins (SNMPs). Notably, a new group of lepidopteran SNMPs (SNMP3 group) was found for the first time in S. exigua, and confirmed in four other moth species. Further, reverse transcription PCR (RT-PCR) and quantitative real time PCR (qPCR) were employed respectively to validate the sequences and determine the expression patterns of 69 identified chemosensory genes regarding to sexes, tissues and stages. Results showed that 67 of these genes could be detected and reconstructed in at least one tissue tested. Further, 60 chemosensory genes were expressed in adult antennae and 52 in larval heads with the antennae, whereas over half of the genes were also detected in non-olfactory tissues like egg and thorax. Particularly, S. exigua OBP2 showed a predominantly larval head-biased expression, and functional studies further indicated its potentially olfactory roles in guiding food searching of larvae. This work suggests functional diversities of S. exigua chemosensory genes and could greatly facilitate the understanding of olfactory system in S. exigua and other lepidopteran species. PMID:26221071

  5. Spodoptera littoralis detoxifies neurotoxic 3-nitropropanoic acid by conjugation with amino acids.

    PubMed

    Novoselov, Alexey; Becker, Tobias; Pauls, Gerhard; von Reuß, Stephan H; Boland, Wilhelm

    2015-08-01

    Spodoptera littoralis is a phytophagous generalist. Its host range includes more than 40 plant species, some of which produce 3-nitropropanoic acid (3-NPA), an irreversible inhibitor of mitochondrial succinate dehydrogenase. Growth in larvae fed an artificial diet with a sublethal admixture of 3-NPA (4.2 μmol per g) was slowed significantly, but larvae experienced no increase in mortality. In contrast, larvae injected with 25.2 μmol/g (bodyweight) 3-NPA experienced acute toxicity and death. To study the detoxification mechanism of 3-NPA in S. littoralis, the insect frass was analyzed by HPLC-MS. Comparative analysis of 3-NPA-treated and -untreated control samples using HR-MS(2) revealed a group of differential signals that were identified as amino acid amides of 3-NPA with glycine, alanine, serine, and threonine. When sublethal amounts of stable isotope-labeled 3-NPA were injected into a larva's hemolymph, 3-NPA amino acid conjugates were identified as putative detoxification products. Bioassays with synthetic standards confirmed that the toxicity of the amides was negligible in comparison to the toxicity of free 3-NPA, demonstrating that amino acid conjugation in S. littoralis represents an efficient way to detoxify 3-NPA. Furthermore, biosynthetic studies using crude fractions of the gut tissue indicated that conjugation of 3-NPA with amino acids occurs in epithelial cells of the insect's gut. Taken together, these results suggest that the detoxification of 3-NPA in S. littoralis proceeds via conjugation to specific amino acids within the epithelial cells followed by export of the nontoxic amino acid conjugates to the hemolymph via as yet uncharacterized mechanisms, most likely involving the Malpighian tubules. PMID:26092560

  6. Structural and Functional Analyses of a Sterol Carrier Protein in Spodoptera litura

    PubMed Central

    Xu, Rui; Zheng, Sichun; He, Hongwu; Wan, Jian; Feng, Qili

    2014-01-01

    Backgrounds In insects, cholesterol is one of the membrane components in cells and a precursor of ecdysteroid biosynthesis. Because insects lack two key enzymes, squalene synthase and lanosterol synthase, in the cholesterol biosynthesis pathway, they cannot autonomously synthesize cholesterol de novo from simple compounds and therefore have to obtain sterols from their diet. Sterol carrier protein (SCP) is a cholesterol-binding protein responsible for cholesterol absorption and transport. Results In this study, a model of the three-dimensional structure of SlSCPx-2 in Spodoptera litura, a destructive polyphagous agricultural pest insect in tropical and subtropical areas, was constructed. Docking of sterol and fatty acid ligands to SlSCPx-2 and ANS fluorescent replacement assay showed that SlSCPx-2 was able to bind with relatively high affinities to cholesterol, stearic acid, linoleic acid, stigmasterol, oleic acid, palmitic acid and arachidonate, implying that SlSCPx may play an important role in absorption and transport of these cholesterol and fatty acids from host plants. Site-directed mutation assay of SlSCPx-2 suggests that amino acid residues F53, W66, F89, F110, I115, T128 and Q131 are critical for the ligand-binding activity of the SlSCPx-2 protein. Virtual ligand screening resulted in identification of several lead compounds which are potential inhibitors of SlSCPx-2. Bioassay for inhibitory effect of five selected compounds showed that AH-487/41731687, AG-664/14117324, AG-205/36813059 and AG-205/07775053 inhibited the growth of S. litura larvae. Conclusions Compounds AH-487/41731687, AG-664/14117324, AG-205/36813059 and AG-205/07775053 selected based on structural modeling showed binding affinity to SlSCPx-2 protein and inhibitory effect on the growth of S. litura larvae. PMID:24454688

  7. A novel cytochrome P450 CYP6AB14 gene in Spodoptera litura (Lepidoptera: Noctuidae) and its potential role in plant allelochemical detoxification

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cytochrome P450 monooxygenases (P450) play a prominent role in the adaptation of insects to host plant chemical defenses. To investigate the potential role of P450s in adaptation of the lepidopteran pest Spodoptera litura to host plant allelochemicals, an expressed sequence data set derived from 6th...

  8. A synthetic cryIC gene, encoding a Bacillus thuringiensis δ-endotoxin, confers Spodoptera resistance in alfalfa and tobacco

    PubMed Central

    Strizhov, Nicolai; Keller, Menachem; Mathur, Jaideep; Koncz-Kálmán, Zsuzsanna; Bosch, Dirk; Prudovsky, Evgenia; Schell, Jeff; Sneh, Baruch; Koncz, Csaba; Zilberstein, Aviah

    1996-01-01

    Spodoptera species, representing widespread polyphagous insect pests, are resistant to Bacillus thuringiensis δ-endotoxins used thus far as insecticides in transgenic plants. Here we describe the chemical synthesis of a cryIC gene by a novel template directed ligation–PCR method. This simple and economical method to construct large synthetic genes can be used when routine resynthesis of genes is required. Chemically phosphorylated adjacent oligonucleotides of the gene to be synthesized are assembled and ligated on a single-stranded, partially homologous template derived from a wild-type gene (cryIC in our case) by a thermostable Pfu DNA ligase using repeated cycles of melting, annealing, and ligation. The resulting synthetic DNA strands are selectively amplified by PCR with short specific flanking primers that are complementary only to the new synthetic DNA. Optimized expression of the synthetic cryIC gene in alfalfa and tobacco results in the production of 0.01–0.2% of total soluble proteins as CryIC toxin and provides protection against the Egyptian cotton leafworm (Spodoptera littoralis) and the beet armyworm (Spodoptera exigua). To facilitate selection and breeding of Spodoptera-resistant plants, the cryIC gene was linked to a pat gene, conferring resistance to the herbicide BASTA. PMID:8986755

  9. Cloning of the heat shock protein 90 and 70 genes from the beet armyworm, Spodoptera exigua, and expression characteristics in relation to thermal stress and development

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Two full-length complementary DNAs (cDNAs) of heat shock protein (HSP) genes (Se-hsp90 and Se-hsp70) were cloned from the beet armyworm, Spodoptera exigua, and their expression was investigated in relation to cold shock, heat shock, and development. The open reading frames of Se-hsp90 and Sehsp70 ar...

  10. EFFECTS OF DROUGHT STRESSED COTTON, GOSSYPIUM HIRSUTUM L., ON BEET ARMYWORM, SPODOPTERA EXIGUA (HUBNER), OVIPOSITION, AND LARVAL FEEDING PREFERENCES AND GROWTH

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The beet armyworm, Spodoptera exigua (Hubner), has been reported to oviposit more on drought stressed than on non-stressed cotton plants. Using potted cotton plants in cages, this study demonstrated that beet armyworms deposited 3.3, 4.6, and 2.3 times more (P less than or equal to 0.05) eggs on co...

  11. SUBMITTED TO NEW JOURNAL (12/06/2002): EFFECT OF PEANUT PLANT FUNGAL INFECTION ON OVIPOSITION PREFERENCE BY SPODOPTERA EXIGUA AND ON HOST SEARCHING BEHAVIOR BY COTESIA MARGINIVENTRIS

    Technology Transfer Automated Retrieval System (TEKTRAN)

    (1.) In the present study we tested the effect of peanut, Arachis hypogaea L. (Leguminosae), stem infection by the white mold fungus, Sclerotium rolfsii Saccodes (Basidiomycetes), on the oviposition preference of beet armyworms (BAW), Spodoptera exigua Hübner (Lepidoptera:Noctuidae) and on the host...

  12. Histopathological effects and determination of the putative receptor of Bacillus thuringiensis Cry1Da toxin in Spodoptera littoralis midgut.

    PubMed

    BenFarhat-Touzri, Dalel; Saadaoui, Marwa; Abdelkefi-Mesrati, Lobna; Saadaoui, Imen; Azzouz, Hichem; Tounsi, Slim

    2013-02-01

    Bacillus thuringiensis subsp. aizawai strain HD133, known by its effectiveness against Spodoptera species, produces many insecticidal proteins including Cry1Ab, Cry1Ca and Cry1Da. In the present study, the insecticidal activity of Cry1Da against Spodoptera littoralis was investigated. It showed toxicity with an LC(50) of 224.4 ng/cm(2) with 95% confidence limits of (178.61-270.19) and an LC(90) of 467.77 ng/cm(2) with 95% confidence limits of (392.89-542.65). The midgut histopathology of Cry1Da fed larvae showed vesicle formation in the apical region, vacuolization and destruction of epithelial cells. Biotinylated-activated Cry1Da toxin bound protein of about 65 kDa on blots of S. littoralis brush border membrane preparations. This putative receptor differs in molecular size from those recognized by Cry1C and Vip3A which are active against this polyphagous insect. This difference in midgut receptors strongly supports the use of Cry1Da as insecticidal agent, particularly in case of Cry and/or Vip-resistance management. PMID:23220238

  13. CELLULAR PATHOLOGY OF A GRANULOSIS VIRUS INFECTION

    EPA Science Inventory

    Nuclear and cytoplasmic ultrastructural changes were examined in Spodoptera frugiperda (SF) larval fat body cells infected with granulosis virus (GV). Soon after infection necleocapsidlike structures were observed within the nucleus associated with nuclear pores. The earliest cel...

  14. CO-OCCLUSION AND PERSISTENCE OF A BACULOVIRUS MUTANT LACKING THE POLYHEDRIN GENE

    EPA Science Inventory

    A co-occlusion process was evaluated as a commercially and ecologically acceptable strategy for the development of genetically improved baculovirus insecticides. oinfection of Spodoptera frugiperda (IPLB-SF-21) tissue culture cells with Aucographa californica nuclear polyhedrosis...

  15. Differential activity of multiple saponins against omnivorous insects with varying feeding preferences

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A variety of saponin glycosides and aglycones from seven different plant families (Aquifoliaceae, Asparagaceae, Caryophyllaceae, Dioscoreaceae, Leguminosae, Rosaceae, Sapindaceae) were tested against the corn earworm, Helicoverpa zea, and the fall armyworm, Spodoptera frugiperda. The corn earworm fe...

  16. Two subclasses of odorant-binding proteins in Spodoptera exigua display structural conservation and functional divergence.

    PubMed

    Liu, N-Y; Yang, F; Yang, K; He, P; Niu, X-H; Xu, W; Anderson, A; Dong, S-L

    2015-04-01

    Although many studies on lepidopteran pheromone-binding proteins (PBPs)/ general odorant-binding proteins (GOBPs) have been reported, the functional differentiation within and between the two odorant-binding protein (OBP) subclasses is still elusive. Here we conducted a comparative study on three SexiPBPs and two SexiGOBPs in Spodoptera exigua. Results showed that all five SexiPBP/GOBP genes have the same intron numbers and conserved exon/intron splice sites. Reverse transcription PCR results showed that these five SexiPBP/GOBPs were primarily expressed in antennae of both sexes and some were also detected in other tissues. Further, quantitative real-time PCR showed that five SexiPBP/GOBPs had different sex-biased expression patterns, with PBP1 being highly male-biased (5.96-fold difference) and PBP3 slightly female-biased (2.43-fold difference), while PBP2 and two GOBPs were approximately sex-equivalent (the absolute value<1.90-fold difference). Binding assays showed that all three SexiPBPs could bind all six sex pheromone components, but SexiPBP1 had much higher affinities [dissociation constant (Ki ) <1.10 μM] than did the other two SexiPBPs (Ki  >1.20 μM). Very intriguingly, SexiGOBP2 displayed even stronger binding to five sex pheromone components (Ki  <0.40 μM) than SexiPBP1. In contrast, SexiGOBP1 only exhibited weak binding to three alcohol-pheromone components. Similar results were obtained for tested pheromone analogues. In addition, each of SexiPBP/GOBPs selectively bound some plant odorants with considerable affinities (Ki  <10.0 μM). Taken together, of the three SexiPBPs, SexiPBP1 may play the most important role in female sex pheromone reception, and additionally all three SexiPBPs can detect some plant odorants, while SexiGOBP2 may be involved in the detection of female sex pheromones in addition to plant odorants. The results strongly suggest functional differentiation within and between the two OBP sub-classes. PMID:25345813

  17. The cytotoxicology of momordicins I and II on Spodoptera litura cultured cell line SL-1.

    PubMed

    Liu, Huan; Wang, Guo-Cai; Zhang, Mao-Xin; Ling, Bing

    2015-07-01

    Momordicin I and II are secondary metabolites from bitter melon (Momordica charantia L.) that are toxic to the Spodoptera litura ovary cell line (SL-1 cell). Both momordicin I and II significantly inhibited SL-1 cells proliferation. IC50 values after 24 h were 8.35 and 82.31 µg/mL, 6.11 and 77.49 µg/mL for 36 h, 4.93 and 49.42 µg/mL for 48 h for cells treated by momordicin I and II, respectively. IC50 values of the azadirachtin A control were 149.63, 54.54 and 23.66 µg/mL at 24, 36 and 48 h respectively, indicating that the cytotoxicity of momordicin I was significantly higher than that of momordicin II and azadirachtin A. Using inverted phase contrast microscopy we found that after 24 h exposure to momordicin I and II, cell shapes changed to circular, swelling increased, adherence ability declined and the cellular membrane bubbled. After 48 h exposure to momordicin I, most cells were suspended and dead; vacuole deformation and cytoplasm leakage indicated that momordicin I was more toxic to the cytoskeleton than momordicin II. Cells treated with momordicin I and II inhibited glucose absorption by 23.04 and 13.38% after 48 h and 47.60 and 20.92% after 60 h. Flow cytometry analysis suggested that SL-1 cells treated with momordicin I and II dramatically accumulated during the G2/M phase of the cell cycle, and total cell protein content increased by 56.93 and 35.81% respectively after 48 h treatment. Following treatment with momordicin I and II the karyotheca dissolved, the chromatin condensed abnormally and the nucleoli were damaged, migrated, or disappeared. The PI fluorescent value by FCM showed that the relative fluorescent intensity of SL-1 cells induced by momordicin I and II increased to 521.45 and 370.17, higher than 135.04 induced by control group treatment for 48 h. This indicated significant damage to the cytomembrane. Overall, the results demonstrate that suppression of cytoskeletal function, interference of mitotic figures

  18. Eliminating host-mediated effects demonstrates Bt maize producing Cry1F has no adverse effects on the parasitoid Cotesia marginiventris

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The fall armyworm, Spodoptera frugiperda, is an important pest of maize in the United States and many tropical areas in the western hemisphere. In 2001, Herculex I ® (Cry1F) maize was commercially planted in the United States to control Lepidoptera, including S. frugiperda. In 2006, a population of ...

  19. Modulation of the temporal pattern of calling behavior of female Spodoptera littoralis by exposure to sex pheromone.

    PubMed

    Sadek, Medhat M; von Wowern, Germund; Löfstedt, Christer; Rosén, Wen-Qi; Anderson, Peter

    2012-01-01

    We have examined the timing of calling behavior in the female Egyptian cotton leafworm, Spodoptera littoralis and its modification by exposure to sex pheromone. The calling rhythm of the female moth was found to be circadian, persistent for at least 4 days once it has been entrained, and could be phase shifted by altering the light:dark regime. We also found that female exposure to pheromone affected the rate and duration of calling. A brief exposure to pheromone gland extract increased the proportion of females calling in a constant dim light and this effect persisted for at least 2 days. In response to pheromone exposure, significantly more females also called late into scotophase when most unexposed control females had ceased calling. The adaptive significance of responding to conspecific sex pheromone is discussed. PMID:22001286

  20. Purification and characterization of NADPH--cytochrome c reductase from the midgut of the southern armyworm (Spodoptera eridania).

    PubMed

    Crankshaw, D L; Hetnarski, K; Wilkinson, C F

    1979-09-01

    1. NADPH-cytochrome c reductase was solubilized with bromelain and purified about 400-fold from sucrose/pyrophosphate-washed microsomal fractions from southern armyworm (Spodoptera eridania) larval midguts. 2. The enzyme has a mol.wt. of 70 035 +/- 1300 and contained 2 mol of flavin/mol of enzyme consisting of almost equimolar amounts of FMN and FAD. 3. Aerobic titration of the enzyme with NADPH caused the formation of a stable half-reduced state at 0.5 mol of NADPH/mol of flavin. 4. Kinetic analysis showed that the reduction of cytochrome c proceeded by a Bi Bi Ping Pong mechanism. 5. Apparent Km values for NADPH and cytochrome c and Ki values for NADP+ and 2'-AMP were considerably higher for the insect reductase than for the mammalian liver enzyme. 6. These are discussed in relation to possible differences in the active sites of the enzymes. PMID:117798

  1. Purification and characterization of NADPH--cytochrome c reductase from the midgut of the southern armyworm (Spodoptera eridania).

    PubMed Central

    Crankshaw, D L; Hetnarski, K; Wilkinson, C F

    1979-01-01

    1. NADPH-cytochrome c reductase was solubilized with bromelain and purified about 400-fold from sucrose/pyrophosphate-washed microsomal fractions from southern armyworm (Spodoptera eridania) larval midguts. 2. The enzyme has a mol.wt. of 70 035 +/- 1300 and contained 2 mol of flavin/mol of enzyme consisting of almost equimolar amounts of FMN and FAD. 3. Aerobic titration of the enzyme with NADPH caused the formation of a stable half-reduced state at 0.5 mol of NADPH/mol of flavin. 4. Kinetic analysis showed that the reduction of cytochrome c proceeded by a Bi Bi Ping Pong mechanism. 5. Apparent Km values for NADPH and cytochrome c and Ki values for NADP+ and 2'-AMP were considerably higher for the insect reductase than for the mammalian liver enzyme. 6. These are discussed in relation to possible differences in the active sites of the enzymes. Images Fig. 3. PMID:117798

  2. Influence ofAmaranthus hybridus L. allelochemics on oviposition behavior ofSpodoptera exigua andS. eridania (Lepidoptera: Noctuidae).

    PubMed

    Mitchell, E R; Heath, R R

    1985-05-01

    Common pigweed,Amaranthus hybridus L., is a favorite host of the beet army worm (BAW),Spodoptera exigua L. Chemicals extracted from homogenized pigweed with distilled water, ethanol, or dichloromethane and sprayed back on pigweed deterred oviposition by the BAW. Similarly, water extracts of frass from conspecific larvae or southern armyworm (SAW) larvae,S. eridania (Cramer), fed pigweed leaves and sprayed back on pigweed plants also deterred BAW oviposition, thus confirming that deterrence was due to plant allelochemics rather than specific compounds associated with the metabolic or excretory products of the larvae. Confirmation of the presence of oviposition-deterring chemicals in pigweed was used to explain a previously observed seasonal displacement of BAW by SAW on pigweed in the field. PMID:24310126

  3. Effect of antibiotic on survival and development of Spodoptera litura (Lepidoptera: Noctuidae) and its gut microbial diversity.

    PubMed

    Thakur, A; Dhammi, P; Saini, H S; Kaur, S

    2016-06-01

    Addition of antibiotics to artificial diets of insects is a key component in the rearing of insects in the laboratory. In the present study an antimicrobial agent, streptomycin sulphate was tested for its influence on survival and fitness of Spodoptera litura (Fabricus) (Lepidoptera: Noctuidae) as well as its gut microbial diversity. The antibiotic did not adversely affect the survival of S. litura. Faster growth of larvae was recorded on diet amended with different concentrations of streptomycin sulphate (0.03, 0.07 and 0.15%) as compared to diet without streptomycin sulphate. The overall activity of various digestives enzymes increased on S+ diet while the activity of detoxifying enzymes significantly decreased. In addition, alteration in microbial diversity was found in the gut of S. litura larvae fed on diet supplemented with antibiotic (S+) and without antibiotic (S-). PMID:26907537

  4. Radar observations of the autumn migration of the beet armyworm Spodoptera exigua (Lepidoptera: Noctuidae) and other moths in northern China.

    PubMed

    Feng, H-Q; Wu, K-M; Cheng, D-F; Guo, Y-Y

    2003-04-01

    The autumn return migration of the beet armyworm Spodoptera exigua Hübner and other insects in northern China was observed with radar and with a simultaneously-operated searchlight trap and ground light-trap at a site in Langfang, near Beijing. The searchlight trap was found to be an efficient tool for trapping migrants and, operated alongside a ground light-trap, could distinguish migrant from locally-flying species. It was confirmed that S. exigua and some other species were high-altitude nocturnal windborne migrants during September and October in northern China. Maximum density of moths typically occurred below 500 m, and strong layering was often observed at about 200 m above ground level in airflows that would carry the moths towards the south. Descent of S. exigua in the vicinity of the radar site in late September was often associated with rain. PMID:12699532

  5. Morphological and molecular characterization of a new microsporidian (Protozoa: Microsporidia) isolated from Spodoptera litura (Fabricius) (Lepidoptera: Noctuidae).

    PubMed

    Johny, S; Kanginakudru, S; Muralirangan, M C; Nagaraju, J

    2006-06-01

    A microsporidium was isolated from larvae of Spodoptera litura (Fabricius) collected from Tamil Nadu, India. This microsporidian species is monomorphic, disporous and develops in direct contact with the cytoplasm of the host cell. The nuclear configuration of merogonic and sporogonic stages was diplokaryotic. The merogonic proliferative stage was unusual that normal development with 1, 2 and 4 binucleated forms were common, while large multinucleate meronts containing 8 and 12 small compact horseshoe-like diplokaryotic nuclei were also observed. The fresh spores were typically ovocylindrical in shape, with a mean size of 3.91 x 1.91 microm and the polar filament length was approximately 90 microm. Infection was systemic with mature spores produced in the midgut, nervous tissue, muscles, labial glands, gonads, tracheae, epidermis, Malpighian tubules and, most extensively, fat body tissues. The new isolate was highly pathogenic to S. litura larvae. Host specificity tests performed on 37 non-target hosts of 5 different insect orders revealed that the new isolate is pathogenic only to lepidopteran insects. We sequenced the 16S small subunit rRNA (SSU rRNA) gene of the isolate and compared it with 72 non-redundant microsporidian sequences from the GenBank. Based on the light microscopic studies and phylogenetic analyses, the new isolate is assigned to the genus Nosema. Significant differences in the SSU rRNA sequence were identified when compared with the type species Nosema bombycis and other closely related species viz., Nosema spodopterae. Structural differences were also observed in the 16S SSU rRNA between the new isolate and the two above-mentioned microsporidian pathogens. We conclude that the microsporidian isolate reported here is distinctly different from the other known species and is likely to be a new species. PMID:16469201

  6. MOLECULAR CHARACTERIZATION OF AUTOPHAGY-RELATED GENE 5 FROM Spodoptera exigua AND EXPRESSION ANALYSIS UNDER VARIOUS STRESS CONDITIONS.

    PubMed

    Liu, Kai-Yu; Xia, Yu-Qian; Zhou, Jing; Chen, Zu-Wen; Lu, Dandan; Zhang, Ning-Zhao; Liu, Xu-Sheng; Ai, Hui; Zhou, Li-Lin

    2016-08-01

    Autophagy is not only involved in development, but also has been proved to attend immune response against invading pathogens. Autophagy protein 5 (ATG5) is an important autophagic protein, which plays a crucial role in autophagosome elongation. Although ATG5 has been well studied in mammal, yeast, and Drosophila, little is known about ATG5 in lepidopteran insects. We cloned putative SeAtg5 gene from Spodoptera exigua larvae by the rapid amplification of cDNA ends method, and its characteristics and the influences of multiple exogenous factors on its expression levels were then investigated. The results showed that the putative S. exigua SeATG5 protein is highly homologous to other insect ATG5 proteins, which has a conserved Pfm domain and multiple phosphorylation sites. Next, fluorescence microscope observation showed that mCherry-SeATG5 was distributed in both nucleus and cytoplasm of Spodoptera litura Sl-HP cells and partially co-localized with BmATG6-GFP, but it almost has no significant co-localization with GFP-HaATG8. Then, the Western blot analysis demonstrated that GFP-SeATG5 conjugated with ATG12. Moreover, real-time PCR revealed that its expression levels significantly increased at the initiation of pupation and the stage of adult. In addition, the expression levels of SeAtg5 can be enhanced by the starvation, UV radiation, and infection of baculovirus and bacterium. However, the expression levels of SeAtg5 decreased at 24 h post treatments in all these treatments except in starvation. These results suggested that SeATG5 might be involved in response of S. exigua under various stress conditions. PMID:27226059

  7. Effectiveness of transgenic Bt cottons against noctuids in the Lower Rio Grande Valley of Texas

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Evaluations of the comparative efficacy of Bacillus thuringiensis Bollgard, Bollgard II and non-Bt traits expressing cottons for control of the noctuid complex composed of bollworms, Helicoverpa zea (Boddie), fall armyworms, Spodoptera frugiperda (J.E. Smith), beet armyworms, Spodoptera exigua (Hübn...

  8. Naturally Occurring Deletion Mutants Are Parasitic Genotypes in a Wild-Type Nucleopolyhedrovirus Population of Spodoptera exigua

    PubMed Central

    Muñoz, Delia; Castillejo, Juan I.; Caballero, Primitivo

    1998-01-01

    A wild-type nucleopolyhedrovirus (NPV) isolate from Spodoptera exigua from Florida (Se-US2) is a variant of the SeMNPV type strain since it has a unique DNA profile but is closely related to other known geographical isolates of SeMNPV. It consists of several genotypic variants, of which seven were identified in a Se-US2 virus stock by a modification of the in vivo cloning method developed by Smith and Crook (Virology 166:240–244, 1988). The US2A variant was the most prevalent genotype, and it was designated the prototype Se-US2 variant, while four of the variants (US2B, US2D, US2F, and US2H) were found at low frequency. US2C and US2E were also very abundant, and their diagnostic bands were easily observed in wild-type isolate restriction endonuclease patterns. The analysis of each variant, compared to the prototype US2A, showed that US2B and US2H presented minor differences, while US2D and US2F contained slightly larger insertions or deletions. Variants US2C and US2E contained major deletions of 21.1 and 14 kb, respectively, mapping at the same genomic region (between 14.5 and 30.2 map units [m.u.] and between 12.8 and 23 m.u., respectively). This is the first report of such deletion mutants in a natural baculovirus population. Variants US2A, US2B, US2D, US2F, and US2H were isolated as pure genotypes, but we failed to clone US2C and US2E in vivo. When these two variants appeared without apparent contamination with any other variant, they lost their pathogenicity for Spodoptera exigua larvae. A further biological characterization showed evidence that these two naturally occurring deletion mutants act as parasitic genotypes in the virus population. Bioassay data also demonstrated that pure US2A is significantly more pathogenic against second-instar S. exigua larvae than the wild-type isolate. The need for precise genotypic characterization of a baculovirus prior to its development as a bioinsecticide is discussed. PMID:9797293

  9. Portable exhausters POR-004 SKID B, POR-005 SKID C, POR-006 SKID D storage plan

    SciTech Connect

    Nelson, O.D.

    1997-09-04

    This document provides a storage plan for portable exhausters POR-004 SKID B, POR-005 SKID C, AND POR-006 SKID D. The exhausters will be stored until they are needed by the TWRS (Tank Waste Remediation Systems) Saltwell Pumping Program. The storage plan provides criteria for portable exhauster storage, periodic inspections during storage, and retrieval from storage.

  10. Development of insect resistant maize plants expressing a chitinase gene from the cotton leaf worm, Spodoptera littoralis.

    PubMed

    Osman, Gamal H; Assem, Shireen K; Alreedy, Rasha M; El-Ghareeb, Doaa K; Basry, Mahmoud A; Rastogi, Anshu; Kalaji, Hazem M

    2015-01-01

    Due to the importance of chitinolytic enzymes for insect, nematode and fungal growth, they are receiving attention concerning their development as biopesticides or chemical defense proteins in transgenic plants and as microbial biocontrol agents. Targeting chitin associated with the extracellular matrices or cell wall by insect chitinases may be an effective approach for controlling pest insects and pathogenic fungi. The ability of chitinases to attack and digest chitin in the peritrophic matrix or exoskeleton raises the possibility to use them as insect control method. In this study, an insect chitinase cDNA from cotton leaf worm (Spodoptera littoralis) has been synthesized. Transgenic maize plant system was used to improve its tolerance against insects. Insect chitinase transcripts and proteins were expressed in transgenic maize plants. The functional integrity and expression of chitinase in progenies of the transgenic plants were confirmed by insect bioassays. The bioassays using transgenic corn plants against corn borer (Sesamia cretica) revealed that ~50% of the insects reared on transgenic corn plants died, suggesting that transgenic maize plants have enhanced resistance against S. cretica. PMID:26658494

  11. Biodiversity and Activity of the Gut Microbiota across the Life History of the Insect Herbivore Spodoptera littoralis.

    PubMed

    Chen, Bosheng; Teh, Beng-Soon; Sun, Chao; Hu, Sirui; Lu, Xingmeng; Boland, Wilhelm; Shao, Yongqi

    2016-01-01

    Microbes that live inside insects play critical roles in host nutrition, physiology, and behavior. Although Lepidoptera (butterflies and moths) are one of the most diverse insect taxa, their microbial symbionts are little-studied, particularly during metamorphosis. Here, using ribosomal tag pyrosequencing of DNA and RNA, we investigated biodiversity and activity of gut microbiotas across the holometabolous life cycle of Spodoptera littoralis, a notorious agricultural pest worldwide. Proteobacteria and Firmicutes dominate but undergo a structural "metamorphosis" in tandem with its host. Enterococcus, Pantoea and Citrobacter were abundant and active in early-instar, while Clostridia increased in late-instar. Interestingly, only enterococci persisted through metamorphosis. Female adults harbored high proportions of Enterococcus, Klebsiella and Pantoea, whereas males largely shifted to Klebsiella. Comparative functional analysis with PICRUSt indicated that early-instar larval microbiome was more enriched for genes involved in cell motility and carbohydrate metabolism, whereas in late-instar amino acid, cofactor and vitamin metabolism increased. Genes involved in energy and nucleotide metabolism were abundant in pupae. Female adult microbiome was enriched for genes relevant to energy metabolism, while an increase in the replication and repair pathway was observed in male. Understanding the metabolic activity of these herbivore-associated microbial symbionts may assist the development of novel pest-management strategies. PMID:27389097

  12. Development of insect resistant maize plants expressing a chitinase gene from the cotton leaf worm, Spodoptera littoralis

    PubMed Central

    Osman, Gamal H.; Assem, Shireen K.; Alreedy, Rasha M.; El-Ghareeb, Doaa K.; Basry, Mahmoud A.; Rastogi, Anshu; Kalaji, Hazem M.

    2015-01-01

    Due to the importance of chitinolytic enzymes for insect, nematode and fungal growth, they are receiving attention concerning their development as biopesticides or chemical defense proteins in transgenic plants and as microbial biocontrol agents. Targeting chitin associated with the extracellular matrices or cell wall by insect chitinases may be an effective approach for controlling pest insects and pathogenic fungi. The ability of chitinases to attack and digest chitin in the peritrophic matrix or exoskeleton raises the possibility to use them as insect control method. In this study, an insect chitinase cDNA from cotton leaf worm (Spodoptera littoralis) has been synthesized. Transgenic maize plant system was used to improve its tolerance against insects. Insect chitinase transcripts and proteins were expressed in transgenic maize plants. The functional integrity and expression of chitinase in progenies of the transgenic plants were confirmed by insect bioassays. The bioassays using transgenic corn plants against corn borer (Sesamia cretica) revealed that ~50% of the insects reared on transgenic corn plants died, suggesting that transgenic maize plants have enhanced resistance against S. cretica. PMID:26658494

  13. Homologs to Cry toxin receptor genes in a de novo transcriptome and their altered expression in resistant Spodoptera litura larvae.

    PubMed

    Gong, Liang; Wang, Huidong; Qi, Jiangwei; Han, Lanzhi; Hu, Meiying; Jurat-Fuentes, Juan Luis

    2015-07-01

    Insect resistance threatens sustainability of insecticides based on Cry proteins from the bacterium Bacillus thuringiensis (Bt). Since high levels of resistance to Cry proteins involve alterations in Cry-binding midgut receptors, their identification is needed to develop resistance management strategies. Through Illumina sequencing we generated a transcriptome containing 16,161 annotated unigenes for the Oriental leafworm (Spodoptera litura). Transcriptome mining identified 6 contigs with identity to reported lepidopteran Cry toxin receptors. Using PCR we confirmed their expression during the larval stage and compared their quantitative expression in larvae from susceptible and a field-derived Cry1Ca resistant strain of S. litura. Among reduced transcript levels detected for most tested contigs in the Cry1Ca-resistant S. litura larvae, the most dramatic reduction (up to 99%) was detected for alkaline phosphatase contigs. This study significantly expands S. litura transcriptomic resources and provides preliminary identification of putative receptor genes with altered expression in S. litura resistant to Cry1Ca toxin. PMID:25981133

  14. Properties of 5-aminolaevulinate synthetase and its relationship to microsomal mixed-function oxidation in the southern armyworm (Spodoptera eridania).

    PubMed

    Brattsten, L B; Wilkinson, C F

    1975-07-01

    1. Activity of 5-aminolaevulinate synthetase was measured in the midgut and other tissues of the last larval instar of the southern armyworm (Spodoptera eridania Cramer, formerly Prodenia eridania Cramer). 2. Optimum conditions for measuring the activity were established with respect to all variables involved and considerable differences from those reported for mammalian enzyme preparations were found. 3. Maximum activity (20 nmol/h per mg of protein) occurs 18-24 h after the fifth moult and thereafter decreases to trace amounts as the larvae age and approach pupation. 4. Synthetase activity was rapidly induced by oral administration (in the diet) of pentamethylbenzene, phenobarbital, diethyl 1,4-dihydro-2,4,6-trimethylpyridine-3, 5-dicarboxylate, and 2-allyl-2-isopropylacetamide. 5. Puromycin inhibited the induction of synthetase by pentamethylbenzene. 6. Induction of 5-aminolaevulinate synthetase correlated well with the induction of microsomal N-demethylation of p-chloro-N-methylaniline, except for phenobarbital, which induced the microsomal oxidase relatively more than the synthetase. PMID:1004

  15. Properties of 5-aminolaevulinate synthetase and its relationship to microsomal mixed-function oxidation in the southern armyworm (Spodoptera eridania).

    PubMed Central

    Brattsten, L B; Wilkinson, C F

    1975-01-01

    1. Activity of 5-aminolaevulinate synthetase was measured in the midgut and other tissues of the last larval instar of the southern armyworm (Spodoptera eridania Cramer, formerly Prodenia eridania Cramer). 2. Optimum conditions for measuring the activity were established with respect to all variables involved and considerable differences from those reported for mammalian enzyme preparations were found. 3. Maximum activity (20 nmol/h per mg of protein) occurs 18-24 h after the fifth moult and thereafter decreases to trace amounts as the larvae age and approach pupation. 4. Synthetase activity was rapidly induced by oral administration (in the diet) of pentamethylbenzene, phenobarbital, diethyl 1,4-dihydro-2,4,6-trimethylpyridine-3, 5-dicarboxylate, and 2-allyl-2-isopropylacetamide. 5. Puromycin inhibited the induction of synthetase by pentamethylbenzene. 6. Induction of 5-aminolaevulinate synthetase correlated well with the induction of microsomal N-demethylation of p-chloro-N-methylaniline, except for phenobarbital, which induced the microsomal oxidase relatively more than the synthetase. PMID:1004

  16. Nerium oleander indirect leaf photosynthesis and light harvesting reductions after clipping injury or Spodoptera eridania herbivory: high sensitivity to injury.

    PubMed

    Delaney, Kevin J

    2012-04-01

    Variable indirect photosynthetic rate (P(n)) responses occur on injured leaves after insect herbivory. It is important to understand factors that influence indirect P(n) reductions after injury. The current study examines the relationship between gas exchange and chlorophyll a fluorescence parameters with injury intensity (% single leaf tissue removal) from clipping or Spodoptera eridania Stoll (Noctuidae) herbivory on Nerium oleander L. (Apocynaceae). Two experiments showed intercellular [CO(2)] increases but P(n) and stomatal conductance reductions with increasing injury intensity, suggesting non-stomatal P(n) limitation. Also, P(n) recovery was incomplete at 3d post-injury. This is the first report of a negative exponential P(n) impairment function with leaf injury intensity to suggest high N. oleander leaf sensitivity to indirect P(n) impairment. Negative linear functions occurred between most other gas exchange and chlorophyll a fluorescence parameters with injury intensity. The degree of light harvesting impairment increased with injury intensity via lower (1) photochemical efficiency indicated lower energy transfer efficiency from reaction centers to PSII, (2) photochemical quenching indicated reaction center closure, and (3) electron transport rates indicated less energy traveling through PSII. Future studies can examine additional mechanisms (mesophyll conductance, carbon fixation, and cardenolide induction) to cause N. oleander indirect leaf P(n) reductions after injury. PMID:22325884

  17. Assessment of electron beam-induced abnormal development and DNA damage in Spodoptera litura (F.) (Lepidoptera: Noctuidae)

    NASA Astrophysics Data System (ADS)

    Yun, Seung-Hwan; Lee, Seon-Woo; Koo, Hyun-Na; Kim, Gil-Hah

    2014-03-01

    The armyworm, Spodoptera litura (F.) is a polyphagous and important agricultural pest worldwide. In this study, we examined the effect of electron beam irradiation on developmental stages, reproduction, and DNA damage of S. litura. Eggs (0-24 h old), larvae (3rd instar), pupae (3 days old after pupation), and adults (24 h after emergence) were irradiated with electron beam irradiation of six levels between 30 and 250 Gy. When eggs were irradiated with 100 Gy, egg hatching was completely inhibited. When the larvae were irradiated, the larval period was significantly delayed, depending on the doses applied. At 150 Gy, the fecundity of adults that developed from irradiated pupae was entirely inhibited. However, electron beam irradiation did not induce the instantaneous death of S. litura adults. Reciprocal crosses between irradiated and unirradiated moths demonstrated that females were more radiosensitive than males. We also conducted the comet assay immediately after irradiation and over the following 5 days period. Severe DNA fragmentation in S. litura cells was observed just after irradiation and the damage was repaired during the post-irradiation period in a time-dependent manner. However, at more than 100 Gy, DNA damage was not fully recovered.

  18. The utility of camptothecin as a synergist of Bacillus thuringiensis var. kurstaki and nucleopolyhedroviruses against Trichoplusia ni and Spodoptera exigua.

    PubMed

    Sun, Shifeng; Cheng, Zhongshan; Fan, Jing; Cheng, Xinghu; Pang, Yi

    2012-08-01

    We studied the effect of combining microbial pesticides with camptothecin (CPT) on the mortality of two lepidopteran insects: Trichoplusia ni (Hübner) and Spodoptera exigua (Hübner). CPT is an alkaloid that is often used as an anticancer agent. Here, CPT was evaluated as a microbial pesticide synergist of Bacillus thuringiensis (Bt) and insect baculovirus. The toxicity of CPT and its synergistic effects on two microbial pesticides were studied using the diet overlay method. Bioassay results showed that CPT significantly enhances the toxicity of Bt variety kurstaki to S. exigua and T ni. In addition, CPT strongly enhanced the infectivity of Autographa californica (Speyer) multinucleocapsid nucleopolyhedrovirus (AcMNPV) and S. exigua nucleopolyhedrovirus (SeMNPV). Using light microscopy, we found that CPT disrupts the peritrophic membrane of T. ni larvae and severely affects the structure of the midgut, resulting in an abnormal gut lumen morphology. We speculate that CPT increases toxicity by affecting the permeability of the peritrophic membrane. PMID:22928294

  19. Elongation Factor 1β′ Gene from Spodoptera exigua: Characterization and Function Identification through RNA Interference

    PubMed Central

    Zhao, Li-Na; Qin, Zi; Wei, Ping; Guo, Hong-Shuang; Dang, Xiang-Li; Wang, Shi-Gui; Tang, Bin

    2012-01-01

    Elongation factor (EF) is a key regulation factor for translation in many organisms, including plants, bacteria, fungi, animals and insects. To investigate the nature and function of elongation factor 1β′ from Spodoptera exigua (SeEF-1β′), its cDNA was cloned. This contained an open reading frame of 672 nucleotides encoding a protein of 223 amino acids with a predicted molecular weight of 24.04 kDa and pI of 4.53. Northern blotting revealed that SeEF-1β′ mRNA is expressed in brain, epidermis, fat body, midgut, Malpighian tubules, ovary and tracheae. RT-PCR revealed that SeEF-1β′ mRNA is expressed at different levels in fat body and whole body during different developmental stages. In RNAi experiments, the survival rate of insects injected with SeEF-1β′ dsRNA was 58.7% at 36 h after injection, which was significantly lower than three control groups. Other elongation factors and transcription factors were also influenced when EF-1β′ was suppressed. The results demonstrate that SeEF-1β′ is a key gene in transcription in S. exigua. PMID:22942694

  20. Identification of plant semiochemicals and characterization of new olfactory sensory neuron types in a polyphagous pest moth, Spodoptera littoralis.

    PubMed

    Binyameen, Muhammad; Anderson, Peter; Ignell, Rickard; Birgersson, Göran; Razaq, Muhammad; Shad, Sarfraz A; Hansson, Bill S; Schlyter, Fredrik

    2014-10-01

    Phytophagous insects use blends of volatiles released from plants to select hosts for feeding and oviposition. To behaviorally analyze complex blends, we need efficient and selective methods for elucidating neuron types, their ligands, and specificity. Gas chromatography-combined single sensillum recordings (GC-SSRs) from antennal olfactory sensilla of female moth, Spodoptera littoralis revealed 38 physiologically active peaks in the headspace volatile blends from both larvae-damaged cotton plants and lilac flowers. Using GC-combined mass spectrometry, 9 new physiologically active compounds were identified from damaged cotton and 11 from lilac compared with earlier electrophysiological studies using antennae of female S. littoralis. We characterized 14 novel classes of olfactory sensory neurons (OSNs). Among these, we found the first 2 ligands for a frequent type of short trichoid sensillum, for which no ligands were identified earlier. By using GC-SSR, a substantial increase in functional classes of OSNs and active compounds, 40% and 34% more, respectively, compared with recent studies using GC-electroantennogram or SSR using single compounds was detected. Compared with the estimated number of corresponding antennal olfactory receptors, the OSN classes now correspond to 83% of a likely maximum. The many specialist OSNs observed may facilitate behavioral confirmation of key plant volatiles in blends. PMID:25194141

  1. Biodiversity and Activity of the Gut Microbiota across the Life History of the Insect Herbivore Spodoptera littoralis

    PubMed Central

    Chen, Bosheng; Teh, Beng-Soon; Sun, Chao; Hu, Sirui; Lu, Xingmeng; Boland, Wilhelm; Shao, Yongqi

    2016-01-01

    Microbes that live inside insects play critical roles in host nutrition, physiology, and behavior. Although Lepidoptera (butterflies and moths) are one of the most diverse insect taxa, their microbial symbionts are little-studied, particularly during metamorphosis. Here, using ribosomal tag pyrosequencing of DNA and RNA, we investigated biodiversity and activity of gut microbiotas across the holometabolous life cycle of Spodoptera littoralis, a notorious agricultural pest worldwide. Proteobacteria and Firmicutes dominate but undergo a structural “metamorphosis” in tandem with its host. Enterococcus, Pantoea and Citrobacter were abundant and active in early-instar, while Clostridia increased in late-instar. Interestingly, only enterococci persisted through metamorphosis. Female adults harbored high proportions of Enterococcus, Klebsiella and Pantoea, whereas males largely shifted to Klebsiella. Comparative functional analysis with PICRUSt indicated that early-instar larval microbiome was more enriched for genes involved in cell motility and carbohydrate metabolism, whereas in late-instar amino acid, cofactor and vitamin metabolism increased. Genes involved in energy and nucleotide metabolism were abundant in pupae. Female adult microbiome was enriched for genes relevant to energy metabolism, while an increase in the replication and repair pathway was observed in male. Understanding the metabolic activity of these herbivore-associated microbial symbionts may assist the development of novel pest-management strategies. PMID:27389097

  2. Elevated atmospheric carbon dioxide concentration affects interactions between Spodoptera exigua (Lepidoptera: Noctuidae) larvae and two host plant species outdoors

    SciTech Connect

    Caulfield, F.; Bunce, J.A. )

    1994-08-01

    Beet armyworm, Spodoptera exigua (Huebner), larvae were placed on sugarbeet (Beta vulgaris L.) and pigweed (Amaranthus hybridus L.) plants in outdoor chambers in which the plants were growing at either the ambient ([approximately] 350 [mu]l liter[sup [minus]1]) or ambient plus 350 [mu]l liter[sup [minus]1] ([approximately] 700 [mu]l liter[sup [minus]1]) carbon dioxide concentration. A series of experiments was performed to determine if larvae reduced plant growth differently at the two carbon dioxide concentrations in either species and if the insect growth or survival differed with carbon dioxide concentration. Leaf nitrogen, water, starch, and soluble carbohydrate contents were measured to assess carbon dioxide concentration effects on leaf quality. Insect feeding significantly reduced plant growth in sugarbeet plants at 350 [mu]l liter[sup [minus]1] but not at 700 [mu]l liter[sup [minus]1] nor in pigweed at either carbon dioxide concentration. Larval survival was greater on sugarbeet plants at the elevated carbon dioxide concentration. Increased survival occurred only if the insects were at the elevated carbon dioxide concentration and consumed leaf material grown at the elevated concentration. Leaf quality was only marginally affected by growth at elevated carbon dioxide concentration in these experiments. The results indicate that in designing experiments to predict effects of elevated atmospheric carbon dioxide concentrations on plant-insect interactions, both plants and insects should be exposed to the experimental carbon dioxide concentrations, as well as to as realistic environmental conditions as possible.

  3. Rhizobacteria activates (+)-δ-cadinene synthase genes and induces systemic resistance in cotton against beet armyworm (Spodoptera exigua).

    PubMed

    Zebelo, Simon; Song, Yuanyuan; Kloepper, Joseph W; Fadamiro, Henry

    2016-04-01

    Gossypol is an important allelochemical produced by the subepidermal glands of some cotton varieties and important for their ability to respond to changing biotic stress by exhibiting antibiosis against some cotton pests. Plant growth-promoting rhizobacteria (PGPR) are root-colonizing bacteria that increase plant growth and often elicit defence against plant pathogens and insect pests. Little is known about the effect of PGPR on cotton plant-insect interactions and the potential biochemical and molecular mechanisms by which PGPR enhance cotton plant defence. Here, we report that PGPR (Bacillus spp.) treated cotton plants showed significantly higher levels of gossypol compared with untreated plants. Similarly, the transcript levels of the genes (i.e. (+)-δ-cadinene synthase gene family) involved in the biosynthesis of gossypol were higher in PGPR-treated plants than in untreated plants. Furthermore, the levels of jasmonic acid, an octadecanoid-derived defence-related phytohormone and the transcript level of jasmonic acid responsive genes were higher in PGPR-treated plants than in untreated plants. Most intriguingly, Spodoptera exigua showed reduced larval feeding and development on PGPR-treated plants. These findings demonstrate that treatment of plants with rhizobacteria may induce significant biochemical and molecular changes with potential ramifications for plant-insect interactions. PMID:26715260

  4. A push-button: Spodoptera exigua oviposition on Nicotiana attenuata dose-independently primes the feeding-induced plant defense.

    PubMed

    Bandoly, Michele; Steppuhn, Anke

    2016-01-01

    Insect oviposition on a plant often precedes the attack by herbivorous larvae. We recently discovered that oviposition by Spodoptera exigua moths on the desert tobacco Nicotiana attenuata primes the induction of 2 defense traits, a phenylpropanoid and activity of protease inhibitors, in response to larval feeding. Oviposition-experienced plants suffer a reduced feeding damage by less and smaller larvae than unexperienced control plants. The increased resistance of oviposition-experienced plants requires the plant's ability to activate its biosynthesis of phenylpropanoids via a Myb transcription factor. Oviposition by S. exigua on N. attenuata is highly variable with respect to the amount, distribution and localization of the eggs on the plant. This raises the question, whether the plant's priming of herbivore defense depends on the egg number and localization. S. exigua moths prefer the oldest leaves for oviposition and yet prime defense-induction in the larval attacked young systemic leaves. Neither the levels of the primed defense traits, nor the affected larval mortality correlate with the number of eggs a plant previously received. This suggests that upon S. exigua oviposition, N. attenuata is shifted - independently of the egg-dose - into a primed state that is responding stronger to the feeding larvae than unprimed plants. PMID:26555313

  5. Identification of microRNAs by small RNA deep sequencing for synthetic microRNA mimics to control Spodoptera exigua.

    PubMed

    Zhang, Yu Liang; Huang, Qi Xing; Yin, Guo Hua; Lee, Samantha; Jia, Rui Zong; Liu, Zhi Xin; Yu, Nai Tong; Pennerman, Kayla K; Chen, Xin; Guo, An Ping

    2015-02-25

    Beet armyworm, Spodoptera exigua, is a major pest of cotton around the world. With the increase of resistance to Bacillus thuringiensis (Bt) toxin in transgenic cotton plants, there is a need to develop an alternative control approach that can be used in combination with Bt transgenic crops as part of resistance management strategies. MicroRNAs (miRNAs), a non-coding small RNA family (18-25 nt), play crucial roles in various biological processes and over-expression of miRNAs has been shown to interfere with the normal development of insects. In this study, we identified 127 conserved miRNAs in S. exigua by using small RNA deep sequencing technology. From this, we tested the effects of 11 miRNAs on larval development. We found three miRNAs, Sex-miR-10-1a, Sex-miR-4924, and Sex-miR-9, to be differentially expressed during larval stages of S. exigua. Oral feeding experiments using synthetic miRNA mimics of Sex-miR-10-1a, Sex-miR-4924, and Sex-miR-9 resulted in suppressed growth of S. exigua and mortality. Over-expression of Sex-miR-4924 caused a significant reduction in the expression level of chitinase 1 and caused abortive molting in the insects. Therefore, we demonstrated a novel approach of using miRNA mimics to control S. exigua development. PMID:25528266

  6. Expression of SNMP1 and SNMP2 genes in antennal sensilla of Spodoptera exigua (Hübner).

    PubMed

    Liu, Chengcheng; Zhang, Jin; Liu, Yang; Wang, Guirong; Dong, Shuanglin

    2014-02-01

    Sensory neuron membrane proteins (SNMPs) are olfactory-specific, two-transmembrane proteins. Previous publications reported that SNMP1 is expressed on the dendrite membrane of pheromone-sensitive neurons in Heliothis virescens and is an essential cofactor for pheromone detection in Drosophila. In this study, we cloned two SNMP genes (GenBank accession nos. JX469106 and JX469107) from the antenna of the beet armyworm moth Spodoptera exigua (Lepidoptera: Hübner). These SNMP genes are classified into two highly conserved subclades, indicating their importance in physiological activity of lepidopteran insects. SexiSNMP1 is antenna-specific in male and female adults, while SexiSNMP2 is antenna-abundant but also expressed in other chemosensory tissues, particularly proboscises and maxillary palps of adults both sexes. In situ hybridization revealed that both SNMPs are broadly expressed in long and short trichoid and basiconic sensilla. We infer that SNMP1 and SNMP2 act in the detection of the sex pheromone and general odorants. PMID:24436214

  7. Dynamics and regulation of glycolysis-tricarboxylic acid metabolism in the midgut of Spodoptera litura during metamorphosis.

    PubMed

    Hu, D; Luo, W; Fan, L F; Liu, F L; Gu, J; Deng, H M; Zhang, C; Huang, L H; Feng, Q L

    2016-04-01

    Significant changes usually take place in the internal metabolism of insects during metamorphosis. The glycolysis-tricarboxylic acid (glycolysis-TCA) pathway is important for energy metabolism. To elucidate its dynamics, the mRNA levels of genes involved in this pathway were examined in the midgut of Spodoptera litura during metamorphosis, and the pyruvate content was quantified. The expression patterns of these genes in response to starvation were examined, and the interaction between protein phosphatase 1 (PP1) and phosphofructokinase (PFK) was studied. The results revealed that the expression or activities of most glycolytic enzymes was down-regulated in prepupae and then recovered in some degree in pupae, and all TCA-related genes were remarkably suppressed in both the prepupae and pupae. Pyruvate was enriched in the pupal midgut. Taken together, these results suggest that insects decrease both glycolysis and TCA in prepupae to save energy and then up-regulate glycolysis but down-regulate TCA in pupae to increase the supply of intermediates for construction of new organs. The expression of all these genes were down-regulated by starvation, indicating that non-feeding during metamorphosis may be a regulator of glycolysis-TCA pathway in the midgut. Importantly, interaction between PP1 and PFK was identified and is suggested to be involved in the regulation of glycolysis. PMID:26683413

  8. Glutathione S-transferase SlGSTE1 in Spodoptera litura may be associated with feeding adaptation of host plants.

    PubMed

    Zou, Xiaopeng; Xu, Zhibin; Zou, Haiwang; Liu, Jisheng; Chen, Shuna; Feng, Qili; Zheng, Sichun

    2016-03-01

    Spodoptera litura is polyphagous pest insect and feeds on plants of more than 90 families. In this study the role of glutathione S-transferase epilson 1 (slgste1) in S. litura in detoxification was examined. This gene was up-regulated in the midgut of S. litura at the transcriptional and protein levels when the insect fed on Brassica juncea or diet containing phytochemicals such as indole-3-carbinol and allyl-isothiocyanate that are metabolic products of sinigrin and glucobrassicin in B. juncea. The SlGSTE1 could catalyze the conjugation of reduced glutathione and indole-3-carbinol and allyl-isothiocyanate, as well as xanthotoxin, which is a furanocoumarin, under in vitro condition. When the expression of Slgste1 in the larvae was suppressed with RNAi, the larval growth and feeding rate were decreased. Furthermore, the up-regulated expression of the SlGSTE1 protein in the midgut of larvae that fed on different host plants was detected by 2-DE and ESI/MS analysis. The feeding adaptation from the most to the least of the larvae for the various host plants was Brassica alboglabra, Brassica linn. Pekinensis, Cucumis sativus, Ipomoea batatas, Arachis hypogaea and Capsicum frutescens. All the results together suggest that Slgste1 is a critical detoxifying enzyme that is induced by phytochmicals in the host plants and, inter alia, may be related to host plant adaptation of S. litura. PMID:26631599

  9. Opposite Effects on Spodoptera littoralis Larvae of High Expression Level of a Trypsin Proteinase Inhibitor in Transgenic Plants1

    PubMed Central

    De Leo, Francesca; Bonadé-Bottino, Michel A.; Ceci, Luigi R.; Gallerani, Raffaele; Jouanin, Lise

    1998-01-01

    This work illustrates potential adverse effects linked with the expression of proteinase inhibitor (PI) in plants used as a strategy to enhance pest resistance. Tobacco (Nicotiana tabacum L. cv Xanthi) and Arabidopsis [Heynh.] ecotype Wassilewskija) transgenic plants expressing the mustard trypsin PI 2 (MTI-2) at different levels were obtained. First-instar larvae of the Egyptian cotton worm (Spodoptera littoralis Boisd.) were fed on detached leaves of these plants. The high level of MTI-2 expression in leaves had deleterious effects on larvae, causing mortality and decreasing mean larval weight, and was correlated with a decrease in the leaf surface eaten. However, larvae fed leaves from plants expressing MTI-2 at the low expression level did not show increased mortality, but a net gain in weight and a faster development compared with control larvae. The low MTI-2 expression level also resulted in increased leaf damage. These observations are correlated with the differential expression of digestive proteinases in the larval gut; overexpression of existing proteinases on low-MTI-2-expression level plants and induction of new proteinases on high-MTI-2-expression level plants. These results emphasize the critical need for the development of a PI-based defense strategy for plants obtaining the appropriate PI-expression level relative to the pest's sensitivity threshold to that PI. PMID:9808744

  10. Modulation of the Ca(2+) signaling pathway by celangulin I in the central neurons of Spodoptera exigua.

    PubMed

    Li, Yuxin; Lian, Xihong; Wan, Yinging; Wang, Duoyi; Chen, Wei; Di, Fengjuan; Wu, Wenjun; Li, Zhengming

    2016-02-01

    Celangulin I is an insecticidal component isolated from Chinese bittersweet Celastrus angulatus. The present study explored the possible effects of celangulin I on the calcium signaling pathway, especially on the L-type Ca(2+) channel and the calcium channels in the endoplasmic reticulum in the central neurons isolated from the third instar larvae of Spodoptera exigua using whole-cell patch-clamp and calcium imaging technique. The results showed that celangulin I could activate the high voltage-gated calcium channel at the concentration of 150μM. The peak currents were increased by 17% of the initial value at the end of the 10-min recording after treated with celangulin I. The rises of intracellular calcium ion concentration ([Ca(2+)]i) in neurons treated by celangulin I showed that the effects of celangulin I were concentration-dependent. Activation of the RyRs by ryanodine decreased the calcium release induced by celangulin I, indicating that celangulin I exerts effect on insect RyRs. Furthermore, we also provided evidence for the first time that celangulin I activates inositol 1,4,5-trisphosphate (IP3) sensitive intracellular calcium release channels in the endoplasmic reticulum third instar larvae neurons of S. exigua. Plausibly, these experimental results can explain the characteristic symptoms of anesthesia and paralysis in celangulin I treated insects. PMID:26821661

  11. Toxicity and differential protein analysis following destruxin A treatment of Spodoptera litura (Lepidoptera: Noctuidae) SL-1 cells.

    PubMed

    Meng, Xiang; Xu, Xiaoxia; Hu, Junjie; Jin, Fengliang; Hu, Qiongbo; Sun, Qiang; Yu, Xiaoqiang; Ren, Shunxiang

    2011-09-15

    The cytotoxicity of a destruxin A (DA) treatment of Spodoptera litura SL-1 cells was investigated. An MTT assay showed that DA was highly toxic to SL-1 cells in a concentration- and time-dependent manner. The IC(50) values of DA, after 24 h and 48 h of treatment, were 17.86 μg/mL and 7.80 μg/mL, respectively. Under inverted phase contrast microscopy (IPCM), it was found that prolonged treatment with DA could induce cell rounding, cellular membrane shrinking, formation of apoptotic bodies, vacuole appearance and cytoplasm leak out. Apoptosis induced by DA was further confirmed by fluorescence microscopy (FM) and flow cytometry (FCM) studies. SL-1 cells entered early apoptosis following a treatment with 2.5 μg/mL DA and entered late apoptosis following a treatment with increasing concentrations of DA. Furthermore, two-dimensional gel electrophoresis (2-DE) analysis was used to identify 22 proteins which were differentially expressed (≥2-fold difference) between control cells and DA-treated cells, and the expression level of these proteins was significantly different between the treated and untreated cells. Our results suggest that these differentially expressed proteins may help explain the diverse biological effects caused by the destruxin A treatment of cells; additionally, some of the identified proteins may have roles in SL-1 cellular proliferation and apoptosis. PMID:21718714

  12. A push-button: Spodoptera exigua oviposition on Nicotiana attenuata dose-independently primes the feeding-induced plant defense

    PubMed Central

    Bandoly, Michele; Steppuhn, Anke

    2016-01-01

    abstract Insect oviposition on a plant often precedes the attack by herbivorous larvae. We recently discovered that oviposition by Spodoptera exigua moths on the desert tobacco Nicotiana attenuata primes the induction of 2 defense traits, a phenylpropanoid and activity of protease inhibitors, in response to larval feeding. Oviposition-experienced plants suffer a reduced feeding damage by less and smaller larvae than unexperienced control plants. The increased resistance of oviposition-experienced plants requires the plant's ability to activate its biosynthesis of phenylpropanoids via a Myb transcription factor. Oviposition by S. exigua on N. attenuata is highly variable with respect to the amount, distribution and localization of the eggs on the plant. This raises the question, whether the plant's priming of herbivore defense depends on the egg number and localization. S. exigua moths prefer the oldest leaves for oviposition and yet prime defense-induction in the larval attacked young systemic leaves. Neither the levels of the primed defense traits, nor the affected larval mortality correlate with the number of eggs a plant previously received. This suggests that upon S. exigua oviposition, N. attenuata is shifted – independently of the egg-dose – into a primed state that is responding stronger to the feeding larvae than unprimed plants. PMID:26555313

  13. Effects of nickel exposure on testicular function, oxidative stress, and male reproductive dysfunction in Spodoptera litura Fabricius.

    PubMed

    Sun, Hongxia; Wu, Wenjing; Guo, Jixing; Xiao, Rong; Jiang, Fengze; Zheng, Lingyan; Zhang, Guren

    2016-04-01

    Nickel is an environmental pollutant that adversely affects the male reproductive system. In the present study, the effects of nickel exposure on Spodoptera litura Fabricius were investigated by feeding larvae artificial diets containing different doses of nickel for three generations. Damage to testes and effects on male reproduction were examined. The amount of nickel that accumulated in the testes of newly emerged males increased as the nickel dose in the diet increased during a single generation. Nickel exposure increased the amount of thiobarbituric acid reactive substances and decreased the amount of glutathione in treatment groups compared with the control. The activity levels of the antioxidant response indices superoxide dismutases, catalase, and glutathione peroxidase in the testes showed variable dose-dependent relationships with nickel doses and duration of exposure. Nickel doses also disrupted the development of the testes by decreasing the weight and volume of testes and the number of eupyrene and apyrene sperm bundles in treatment groups compared with the control. When the nickel-treated males mated with normal females, fecundity was inhibited by the higher nickel doses in all three generations, but fecundity significantly increased during the second generation, which received 5 mg kg(-1) nickel. Hatching rates in all treatments significantly decreased in a dose-dependent manner in the three successive generations. The effects of nickel on these parameters correlated with the duration of nickel exposure. Results indicate assays of testes may be a novel and efficient means of evaluating the effects of heavy metals on phytophagous insects in an agricultural environment. PMID:26807937

  14. Identification and validation of reference genes for gene expression analysis using quantitative PCR in Spodoptera litura (Lepidoptera: Noctuidae).

    PubMed

    Lu, Yanhui; Yuan, Miao; Gao, Xiwu; Kang, Tinghao; Zhan, Sha; Wan, Hu; Li, Jianhong

    2013-01-01

    Reverse transcription quantitative polymerase chain reaction (qRT-PCR) has rapidly become the most sensitive and accurate method for the quantification of gene expression. To facilitate gene expression studies and obtain more accurate qRT-PCR data, normalization relative to stable housekeeping genes is required. These housekeeping genes need to show stable expression under the given experimental conditions for the qRT-PCR results to be accurate. Unfortunately, there are no studies on the stability of housekeeping genes used in Spodoptera litura. In this study, eight candidate reference genes, elongation factor 1 alpha (EF1), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), ribosomal protein L10 (RPL10), ribosomal protein S3 (RPS3), beta actin (ACTB), beta FTZ-F1 (FTZF1), ubiquinol-cytochrome c reductase (UCCR), and arginine kinase (AK), were evaluated for their suitability as normalization genes under different experimental conditions using the statistical software programs, BestKeeper, geNorm and Normfinder, and the comparative ΔCt method. We determined the expression levels of the candidate reference genes for three biotic factors (developmental stage, tissue and population), and four abiotic treatments (temperature, insecticide, food and starvation). The results indicated that the best sets of candidates as reference genes were as follows: GAPDH and UCCR for developmental stages; RPL10, AK and EF1 for different tissues; RPL10 and EF1 for different populations in China; GAPDH and EF1 for temperature-stressed larvae; AK and ACTB for larvae treated with different insecticides; RPL10, GAPDH and UCCR for larvae fed different diets; RPS3 and ACTB for starved larvae. We believe that these results make an important contribution to gene analysis studies in S. litura and form the basis of further research on stable reference genes in S. litura and other organisms. PMID:23874494

  15. Host plant resistance in romaine lettuce affects feeding behavior and biology of Trichoplusia ni and Spodoptera exigua (Lepidoptera: Noctuidae).

    PubMed

    Sethi, Amit; McAuslane, Heather J; Nagata, Russell T; Nuessly, Gregg S

    2006-12-01

    Lettuce quality and yield can be reduced by feeding of several lepidopterous pests, particularly cabbage looper, Trichoplusia ni (Hübner), and beet armyworm, Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae). Host plant resistance to these insects is an environmentally sound adjunct to conventional chemical control. In this study we compared the survival, development, and feeding behavior of cabbage looper and beet armyworm on two romaine lettuce cultivars, resistant 'Valmaine' and susceptible 'Tall Guzmaine'. Larval mortality of both species was significantly higher on resistant Valmaine than on susceptible Tall Guzmaine. The average weight per larva after feeding for 1 wk on Tall Guzmaine plants was 6 times (beet armyworm) and 2 times (cabbage looper) greater than that of larvae feeding on Valmaine plants. Significant reduction in larval growth on Valmaine compared with that on Tall Guzmaine resulted in a 5.9- (beet armyworm) and 2.6-d (cabbage looper) increase in larval duration and almost a 1-d increase in pupal duration. Average pupal and adult weights and successful pupation of cabbage looper and beet armyworm were reduced on Valmaine compared with Tall Guzmaine. The sex ratio of progeny did not deviate from 1:1 when larvae were reared on either Valmaine or Tall Guzmaine. The fecundity of cabbage looper and beet armyworm adults that developed from larvae reared on Valmaine was about one-third that of adults from Tall Guzmaine, but adult longevity did not significantly differ on the two lettuce cultivars. The two insect species showed different feeding preferences for leaves of different age groups on Valmaine and Tall Guzmaine. Cabbage loopers cut narrow trenches on the leaf before actual feeding to block the flow of latex to the intended site of feeding. In contrast, beet armyworms did not trench. The different feeding behavior of the two species on Valmaine may explain the superior performance of cabbage looper compared with beet armyworm. PMID:17195688

  16. Identification and RNA Interference of the Pheromone Biosynthesis Activating Neuropeptide (PBAN) in the Common Cutworm Moth Spodoptera litura (Lepidoptera: Noctuidae).

    PubMed

    Lu, Qin; Huang, Ling-Yan; Chen, Peng; Yu, Jin-Feng; Xu, Jin; Deng, Jian-Yu; Ye, Hui

    2015-06-01

    Spodoptera litura F. is one of the most destructive insect pests of many agricultural crops and notorious for developing insecticide resistance. Developing environmental friendly control methods such as novel pheromone and RNAi-related control strategies is imperative to control this pest. In the present study, the full-length cDNA encoding the diapause hormone and pheromone biosynthesis activating neuropeptide (DH-PBAN) was identified and characterized in S. litura. This 809-bp transcript contains a 573-nucleotide ORF encoding a 191-amino acid protein, from which five putative neuropeptides, including PBAN, DH, and α-, β-, and γ-subesophageal ganglion neuropeptides, were derived. Phylogenetic analysis showed that both the whole protein and each of the five neuropeptides have high similarities to those of DH-PBANs from other insect orders particularly Lepidoptera. Females treated with TKYFSPRLamide (the active core fragment of PBAN) produced significantly more four types of pheromone compounds (A; B; C; D) than controls. RNA interference by injection of PBAN dsRNA significantly reduced the relative expression levels of this gene in adult females (approximately reduced by 60%). As a consequence, females treated with PBAN dsRNA produced significantly less four types of pheromone compounds (A; B; C; D) than controls. These results suggest that PBAN function in activating sex pheromone biosynthesis and the RNAi of DH-PBAN gene can be induced by the injection of dsRNA into the body cavity in S. litura. This study suggests the possibility of novel pheromone-related pest control strategies based on RNAi techniques. PMID:26470263

  17. Deep-tissue confocal imaging of the central projections of ovipositor sensory afferents in the Egyptian cotton leafworm, Spodoptera littoralis.

    PubMed

    Seada, Mervat A; Ghaninia, Majid

    2016-03-01

    The pre-ovipositon behavior of moths is largely dependent upon the cues that a gravid female perceives while assessing potential oviposition sites. Assessment of such sites is accomplished, at least in part, by mechanosensory and gustatory sensilla located on the ovipositor whose sensory neurons project into the terminal abdominal ganglion (TAG). Using anterograde backfill staining, confocal laser scanning microscopy, and three dimensional reconstruction, we traced and analyzed the central projections of the sensory neurons housed in the sensilla located on the ovipositor papillae and explored the neuropilar composition of the TAG in the Egyptian cotton leafworm, Spodoptera littoralis. The TAG consists of three fused neuromeres (6-8th Ner) associated with the 6-8th abdominal segments. Within the TAG, and specifically in the 8th neuromere, four unstructured neuropilar compartments are present; the dorso-ipsilateral motor neuropil (MN), the medio-ipsilateral mechanosensory neuropil (MchN), the medio-ipsilateral small gustatory neuropil (GN), and the medio-contralateral posterior ovipositor glomerulus (Og). The Og appears quite compact, with a hollow core free of terminal arborizations. The MchN is further subdivided into 4 unstructured glomeruli in the 8th neuromere, whose afferents are subsequently extended into 3 glomeruli in the 7th and 6th neuromeres. Few neurites of the Og are populated with large dense varicosities reminiscent of neurosecretory vesicles. Given that all ovipositor nerves converge into a common ganglionic center, the TAG, we assume that this ganglion may be a center for coordination of oviposition behaviors, including movements of the ovipositor during assessment of oviposition substrates and egg laying in S. littoralis. PMID:26774745

  18. CML42-Mediated Calcium Signaling Coordinates Responses to Spodoptera Herbivory and Abiotic Stresses in Arabidopsis1[W][OA

    PubMed Central

    Vadassery, Jyothilakshmi; Reichelt, Michael; Hause, Bettina; Gershenzon, Jonathan; Boland, Wilhelm; Mithöfer, Axel

    2012-01-01

    In the interaction between Arabidopsis (Arabidopsis thaliana) and the generalist herbivorous insect Spodoptera littoralis, little is known about early events in defense signaling and their link to downstream phytohormone pathways. S. littoralis oral secretions induced both Ca2+ and phytohormone elevation in Arabidopsis. Plant gene expression induced by oral secretions revealed up-regulation of a gene encoding a calmodulin-like protein, CML42. Functional analysis of cml42 plants revealed more resistance to herbivory than in the wild type, because caterpillars gain less weight on the mutant, indicating that CML42 negatively regulates plant defense; cml42 also showed increased aliphatic glucosinolate content and hyperactivated transcript accumulation of the jasmonic acid (JA)-responsive genes VSP2 and Thi2.1 upon herbivory, which might contribute to increased resistance. CML42 up-regulation is negatively regulated by the jasmonate receptor Coronatine Insensitive1 (COI1), as loss of functional COI1 resulted in prolonged CML42 activation. CML42 thus acts as a negative regulator of plant defense by decreasing COI1-mediated JA sensitivity and the expression of JA-responsive genes and is independent of herbivory-induced JA biosynthesis. JA-induced Ca2+ elevation and root growth inhibition were more sensitive in cml42, also indicating higher JA perception. Our results indicate that CML42 acts as a crucial signaling component connecting Ca2+ and JA signaling. CML42 is localized to cytosol and nucleus. CML42 is also involved in abiotic stress responses, as kaempferol glycosides were down-regulated in cml42, and impaired in ultraviolet B resistance. Under drought stress, the level of abscisic acid accumulation was higher in cml42 plants. Thus, CML42 might serve as a Ca2+ sensor having multiple functions in insect herbivory defense and abiotic stress responses. PMID:22570470

  19. Functional characterization of SlitPBP3 in Spodoptera litura by CRISPR/Cas9 mediated genome editing.

    PubMed

    Zhu, Guan-Heng; Xu, Jun; Cui, Zhen; Dong, Xiao-Tong; Ye, Zhan-Feng; Niu, Dong-Juan; Huang, Yong-Ping; Dong, Shuang-Lin

    2016-08-01

    Functional gene analysis by using genome editing techniques is limited only in few model insects. Here, we reported an efficient and heritable gene mutagenesis analysis in an important lepidopteran pest, Spodoptera litura, using the CRISPR/Cas9 system. By using this system, we successfully obtained the homozygous S. litura strain by targeting the pheromone binding protein 3 gene (SlitPBP3), which allowed us to elucidate the role of this gene in the olfaction of the female sex pheromones. By co-injection of Cas9 mRNA and sgRNA into S. litura eggs, highly efficient chimera mutation in SlitPBP3 loci was detected both in injected eggs (39.1%) and in the resulting individual moths (87.5%). We used the mutant moths as parents to obtain the G1 offspring and the homozygous mutant strain in G2. The function of SlitPBP3 was explored by Electroantennogram (EAG) recordings with a homozygous mutant strain. The result showed that the EAG responses were significantly decreased in mutant males than in control males when treated with the major sex pheromone component (Z9,E11-14:Ac) and a minor component (Z9-14:Ac) at higher dosages. The results demonstrate that s SlitPBP3 gene plays a minor role in the perception of the female sex pheromones. Furthermore, our study provides a useful methodology with the CRISPR/Cas9 system for gene in vivo functional study, particular for lepidopteran species in which the RNAi approach is not efficient. PMID:27192033

  20. A Comparison of the Olfactory Gene Repertoires of Adults and Larvae in the Noctuid Moth Spodoptera littoralis

    PubMed Central

    Poivet, Erwan; Gallot, Aurore; Montagné, Nicolas; Glaser, Nicolas; Legeai, Fabrice; Jacquin-Joly, Emmanuelle

    2013-01-01

    To better understand the olfactory mechanisms in a lepidopteran pest model species, the cotton leafworm Spodoptera littoralis, we have recently established a partial transcriptome from adult antennae. Here, we completed this transcriptome using next generation sequencing technologies, namely 454 and Illumina, on both adult antennae and larval tissues, including caterpillar antennae and maxillary palps. All sequences were assembled in 77,643 contigs. Their analysis greatly enriched the repertoire of chemosensory genes in this species, with a total of 57 candidate odorant-binding and chemosensory proteins, 47 olfactory receptors, 6 gustatory receptors and 17 ionotropic receptors. Using RT-PCR, we conducted the first exhaustive comparison of olfactory gene expression between larvae and adults in a lepidopteran species. All the 127 candidate olfactory genes were profiled for expression in male and female adult antennae and in caterpillar antennae and maxillary palps. We found that caterpillars expressed a smaller set of olfactory genes than adults, with a large overlap between these two developmental stages. Two binding proteins appeared to be larvae-specific and two others were adult-specific. Interestingly, comparison between caterpillar antennae and maxillary palps revealed numerous organ-specific transcripts, suggesting the complementary involvement of these two organs in larval chemosensory detection. Adult males and females shared the same set of olfactory transcripts, except two male-specific candidate pheromone receptors, two male-specific and two female-specific odorant-binding proteins. This study identified transcripts that may be important for sex-specific or developmental stage-specific chemosensory behaviors. PMID:23565215

  1. The Arabidopsis immune regulator SRFR1 dampens defences against herbivory by Spodoptera exigua and parasitism by Heterodera schachtii.

    PubMed

    Nguyen, Phuong Dung T; Pike, Sharon; Wang, Jianying; Nepal Poudel, Arati; Heinz, Robert; Schultz, Jack C; Koo, Abraham J; Mitchum, Melissa G; Appel, Heidi M; Gassmann, Walter

    2016-05-01

    Plants have developed diverse mechanisms to fine tune defence responses to different types of enemy. Cross-regulation between signalling pathways may allow the prioritization of one response over another. Previously, we identified SUPPRESSOR OF rps4-RLD1 (SRFR1) as a negative regulator of ENHANCED DISEASE SUSCEPTIBILITY1 (EDS1)-dependent effector-triggered immunity against the bacterial pathogen Pseudomonas syringae pv. tomato strain DC3000 expressing avrRps4. The use of multiple stresses is a powerful tool to further define gene function. Here, we examined whether SRFR1 also impacts resistance to a herbivorous insect in leaves and to a cyst nematode in roots. Interestingly, srfr1-1 plants showed increased resistance to herbivory by the beet army worm Spodoptera exigua and to parasitism by the cyst nematode Heterodera schachtii compared with the corresponding wild-type Arabidopsis accession RLD. Using quantitative real-time PCR (qRT-PCR) to measure the transcript levels of salicylic acid (SA) and jasmonate/ethylene (JA/ET) pathway genes, we found that enhanced resistance of srfr1-1 plants to S. exigua correlated with specific upregulation of the MYC2 branch of the JA pathway concurrent with suppression of the SA pathway. In contrast, the greater susceptibility of RLD was accompanied by simultaneously increased transcript levels of SA, JA and JA/ET signalling pathway genes. Surprisingly, mutation of either SRFR1 or EDS1 increased resistance to H. schachtii, indicating that the concurrent presence of both wild-type genes promotes susceptibility. This finding suggests a novel form of resistance in Arabidopsis to the biotrophic pathogen H. schachtii or a root-specific regulation of the SA pathway by EDS1, and places SRFR1 at an intersection between multiple defence pathways. PMID:26310916

  2. Identification and Validation of Reference Genes for Gene Expression Analysis Using Quantitative PCR in Spodoptera litura (Lepidoptera: Noctuidae)

    PubMed Central

    Gao, Xiwu; Kang, Tinghao; Zhan, Sha; Wan, Hu; Li, Jianhong

    2013-01-01

    Reverse transcription quantitative polymerase chain reaction (qRT-PCR) has rapidly become the most sensitive and accurate method for the quantification of gene expression. To facilitate gene expression studies and obtain more accurate qRT-PCR data, normalization relative to stable housekeeping genes is required. These housekeeping genes need to show stable expression under the given experimental conditions for the qRT-PCR results to be accurate. Unfortunately, there are no studies on the stability of housekeeping genes used in Spodoptera litura. In this study, eight candidate reference genes, elongation factor 1 alpha (EF1), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), ribosomal protein L10 (RPL10), ribosomal protein S3 (RPS3), beta actin (ACTB), beta FTZ-F1 (FTZF1), ubiquinol-cytochrome c reductase (UCCR), and arginine kinase (AK), were evaluated for their suitability as normalization genes under different experimental conditions using the statistical software programs, BestKeeper, geNorm and Normfinder, and the comparative ΔCt method. We determined the expression levels of the candidate reference genes for three biotic factors (developmental stage, tissue and population), and four abiotic treatments (temperature, insecticide, food and starvation). The results indicated that the best sets of candidates as reference genes were as follows: GAPDH and UCCR for developmental stages; RPL10, AK and EF1 for different tissues; RPL10 and EF1 for different populations in China; GAPDH and EF1 for temperature-stressed larvae; AK and ACTB for larvae treated with different insecticides; RPL10, GAPDH and UCCR for larvae fed different diets; RPS3 and ACTB for starved larvae. We believe that these results make an important contribution to gene analysis studies in S. litura and form the basis of further research on stable reference genes in S. litura and other organisms. PMID:23874494

  3. Dissimilar Regulation of Antimicrobial Proteins in the Midgut of Spodoptera exigua Larvae Challenged with Bacillus thuringiensis Toxins or Baculovirus

    PubMed Central

    Crava, Cristina M.; Jakubowska, Agata K.; Escriche, Baltasar; Herrero, Salvador; Bel, Yolanda

    2015-01-01

    Antimicrobial peptides (AMPs) and lysozymes are the main effectors of the insect immune system, and they are involved in both local and systemic responses. Among local responses, midgut immune reaction plays an important role in fighting pathogens that reach the insect body through the oral route, as do many microorganisms used in pest control. Under this point of view, understanding how insects defend themselves locally during the first phases of infections caused by food-borne pathogens is important to further improve microbial control strategies. In the present study, we analyzed the transcriptional response of AMPs and lysozymes in the midgut of Spodoptera exigua (Lepidoptera: Noctuidae), a polyphagous pest that is commonly controlled by products based on Bacillus thuringiensis (Bt) or baculovirus. First, we comprehensively characterized the transcripts encoding AMPs and lysozymes expressed in S. exigua larval midgut, identifying 35 transcripts that represent the S. exigua arsenal against microbial infection. Secondly, we analyzed their expression in the midgut after ingestion of sub-lethal doses of two different pore-forming B. thuringiensis toxins, Cry1Ca and Vip3Aa, and the S. exigua nucleopolyhedrovirus (SeMNPV). We observed that both Bt toxins triggered a similar, wide and in some cases high transcriptional activation of genes encoding AMPs and lysozymes, which was not reflected in the activation of the classical systemic immune-marker phenoloxidase in hemolymph. Baculovirus ingestion resulted in the opposed reaction: Almost all transcripts coding for AMPs and lysozymes were down-regulated or not induced 96 hours post infection. Our results shed light on midgut response to different virulence factors or pathogens used nowadays as microbial control agents and point out the importance of the midgut immune response contribution to the larval immunity. PMID:25993013

  4. Eicosanoids up-regulate production of reactive oxygen species by NADPH-dependent oxidase in Spodoptera exigua phagocytic hemocytes.

    PubMed

    Park, Youngjin; Stanley, David W; Kim, Yonggyun

    2015-08-01

    Eicosanoids mediate cellular immune responses in insects, including phagocytosis of invading microbes. Phagocytosis entails two major steps, the internalization of microbes and the subsequent killing of them via formation of reactive oxygen species (ROS). Here, we posed the hypothesis that eicosanoids mediate ROS production by activating NADPH-dependent oxidase (NOX) and tested the idea in the model insect, Spodoptera exigua. A NOX gene (we named SeNOX4) was identified and cloned, yielding a full open reading frame encoding 547 amino acid residues with a predicted molecular weight of 63,410Da and an isoelectric point at 9.28. A transmembrane domain and a large intracellular domain containing NADPH and FAD-binding sites were predicted. Phylogenetic analysis indicated SeNOX4 clusters with other NOX4 genes. SeNOX4 was expressed in all life stages except eggs, and exclusively in hemocytes. Bacterial challenge and, separately, arachidonic acid (AA, a precursor of eicosanoid biosynthesis) injection increased its expression. The internalization step was assessed by counting hemocytes engulfing fluorescence-labeled bacteria. The phagocytic behavior was inhibited by dsRNA suppression of SeNOX4 expression and, separately by dexamethasone (DEX, a specific inhibitor of eicosanoid biosynthesis) treatments. However, injecting AA to dsSeNOX4-treated larvae did not rescue the phagocytic activity. Hemocytic ROS production increased following bacterial challenge, which was sharply reduced in dsSeNOX4-treated, and separately, in DEX-treated larvae. AA partially reversed the suppressed ROS production in dsSeNOX4-treated larvae. Treating larvae with either the ROS-suppressing dsSeNOX4 construct or DEX rendered experimental larvae unable to inhibit bacterial proliferation in their hemocoels. We infer that eicosanoids mediate ROS production during phagocytosis by inducing expression of SeNOX4. PMID:26071791

  5. Nucleopolyhedrovirus infection and/or parasitism by Microplitis pallidipes Szepligeti affect hemocyte apoptosis of Spodoptera exigua (Hübner) larvae.

    PubMed

    Wan, Nian-Feng; Ji, Xiang-Yun; Zhang, Hao; Yang, Jun-Hua; Jiang, Jie-Xian

    2015-11-01

    We determined the effects of parasitism by the endoparasitoid Microplitis pallidipes Szepligeti and/or nucleopolyhedrovirus (NPV) infection on hemocyte apoptosis of Spodoptera exigua (Hübner) larvae. Compared to healthy (control) larvae, larvae that were parasitized, virus-infected, or both all showed a significant increase in hemocyte apoptosis during 48-h observation period. The peaks of hemocyte apoptosis in parasitized, virus-infected and parasitized+infected larvae were at 12, 24 and 48 h after treatment, and were 86.7±1.9, 87.4±3.6 and 76.5±1.6%, respectively. Meanwhile, compared to parasitized larvae, hemocyte apoptosis in jointly parasitized and infected larvae increased by 12.9%, 18.7% and 2.8% at 8, 36 and 48 h respectively, and decreased by 39.0% and 9.1% at 12 and 24h. Compared to virus-infected larvae, hemocyte apoptosis in jointly parasitized and infected larvae increased by 13.4%, 2.4% and 15.3% at 8, 36 and 48 h, respectively, and decreased by 4.0% and 29.9% at 12 and 24h. Our study found that joint and separate parasitism and SeNPV infection induced hemocyte apoptosis of S. exigua larvae. It also revealed that NPV infection promoted host hemocyte apoptosis induced by parasitism at early egg and larval stages of M. pallidipes in host larvae, but inhibited the same effect at late egg stage of M. pallidipes in host larvae, and that parasitism promoted host hemocyte apoptosis induced by NPV infection at early egg and larval stages of M. pallidipes in host larvae, but inhibited the same effect at late egg stage of M. pallidipes in host larvae. PMID:26470677

  6. Dissimilar Regulation of Antimicrobial Proteins in the Midgut of Spodoptera exigua Larvae Challenged with Bacillus thuringiensis Toxins or Baculovirus.

    PubMed

    Crava, Cristina M; Jakubowska, Agata K; Escriche, Baltasar; Herrero, Salvador; Bel, Yolanda

    2015-01-01

    Antimicrobial peptides (AMPs) and lysozymes are the main effectors of the insect immune system, and they are involved in both local and systemic responses. Among local responses, midgut immune reaction plays an important role in fighting pathogens that reach the insect body through the oral route, as do many microorganisms used in pest control. Under this point of view, understanding how insects defend themselves locally during the first phases of infections caused by food-borne pathogens is important to further improve microbial control strategies. In the present study, we analyzed the transcriptional response of AMPs and lysozymes in the midgut of Spodoptera exigua (Lepidoptera: Noctuidae), a polyphagous pest that is commonly controlled by products based on Bacillus thuringiensis (Bt) or baculovirus. First, we comprehensively characterized the transcripts encoding AMPs and lysozymes expressed in S. exigua larval midgut, identifying 35 transcripts that represent the S. exigua arsenal against microbial infection. Secondly, we analyzed their expression in the midgut after ingestion of sub-lethal doses of two different pore-forming B. thuringiensis toxins, Cry1Ca and Vip3Aa, and the S. exigua nucleopolyhedrovirus (SeMNPV). We observed that both Bt toxins triggered a similar, wide and in some cases high transcriptional activation of genes encoding AMPs and lysozymes, which was not reflected in the activation of the classical systemic immune-marker phenoloxidase in hemolymph. Baculovirus ingestion resulted in the opposed reaction: Almost all transcripts coding for AMPs and lysozymes were down-regulated or not induced 96 hours post infection. Our results shed light on midgut response to different virulence factors or pathogens used nowadays as microbial control agents and point out the importance of the midgut immune response contribution to the larval immunity. PMID:25993013

  7. Model and Scenario Variations in Predicted Number of Generations of Spodoptera litura Fab. on Peanut during Future Climate Change Scenario

    PubMed Central

    Srinivasa Rao, Mathukumalli; Swathi, Pettem; Rama Rao, Chitiprolu Anantha; Rao, K. V.; Raju, B. M. K.; Srinivas, Karlapudi; Manimanjari, Dammu; Maheswari, Mandapaka

    2015-01-01

    The present study features the estimation of number of generations of tobacco caterpillar, Spodoptera litura. Fab. on peanut crop at six locations in India using MarkSim, which provides General Circulation Model (GCM) of future data on daily maximum (T.max), minimum (T.min) air temperatures from six models viz., BCCR-BCM2.0, CNRM-CM3, CSIRO-Mk3.5, ECHams5, INCM-CM3.0 and MIROC3.2 along with an ensemble of the six from three emission scenarios (A2, A1B and B1). This data was used to predict the future pest scenarios following the growing degree days approach in four different climate periods viz., Baseline-1975, Near future (NF) -2020, Distant future (DF)-2050 and Very Distant future (VDF)—2080. It is predicted that more generations would occur during the three future climate periods with significant variation among scenarios and models. Among the seven models, 1–2 additional generations were predicted during DF and VDF due to higher future temperatures in CNRM-CM3, ECHams5 & CSIRO-Mk3.5 models. The temperature projections of these models indicated that the generation time would decrease by 18–22% over baseline. Analysis of variance (ANOVA) was used to partition the variation in the predicted number of generations and generation time of S. litura on peanut during crop season. Geographical location explained 34% of the total variation in number of generations, followed by time period (26%), model (1.74%) and scenario (0.74%). The remaining 14% of the variation was explained by interactions. Increased number of generations and reduction of generation time across the six peanut growing locations of India suggest that the incidence of S. litura may increase due to projected increase in temperatures in future climate change periods. PMID:25671564

  8. A Transcriptome Analysis Suggests Apoptosis-Related Signaling Pathways in Hemocytes of Spodoptera litura After Parasitization by Microplitis bicoloratus

    PubMed Central

    Zhang, Yan; Yu, Dongshuai; Yang, Minjun; Yang, Yang; Hu, Jiansheng; Luo, Kaijun

    2014-01-01

    Microplitis bicoloratus parasitism induction of apoptotic DNA fragmentation of host Spodoptera litura hemocytes has been reported. However, how M. bicoloratus parasitism regulates the host signaling pathways to induce DNA fragmentation during apoptosis remains unclear. To address this question, we performed a new RNAseq-based comparative analysis of the hemocytes transcriptomes of non-parasitized and parasitized S. litura. We were able to assemble a total of more than 11.63 Gbp sequence, to yield 20,571 unigenes. At least six main protein families encoded by M. bicoloratus bracovirus are expressed in the parasitized host hemocytes: Ankyrin-repeat, Ben domain, C-type lectin, Egf-like and Mucin-like, protein tyrosine phosphatase. The analysis indicated that during DNA fragmentation and cell death, 299 genes were up-regulated and 2,441 genes were down-regulated. Data on five signaling pathways related with cell death, the gap junctions, Ca2+, PI3K/Akt, NF-κB, ATM/p53 revealed that CypD, which is involved in forming a Permeability Transition Pore Complex (PTPC) to alter mitochondrial membrane permeabilization (MMP), was dramatically up-regulated. The qRT-PCR also provided that the key genes for cell survival were down-regulated under M. bicoloratus parasitism, including those encoding Inx1, Inx2 and Inx3 of the gap junction signaling pathway, p110 subunit of the PI3K/Akt signaling pathway, and the p50 and p65 subunit of the NF-κB signaling pathway. These findings suggest that M. bicoloratus parasitism may regulate host mitochondria to trigger internucleosomal DNA fragmentation. This study will facilitate the identification of immunosuppression-related genes and also improves our understanding of molecular mechanisms underlying polydnavirus-parasitoid-host interaction. PMID:25350281

  9. Response of the common cutworm Spodoptera litura to lead stress: changes in sex ratio, Pb accumulations, midgut cell ultrastructure.

    PubMed

    Shu, Yinghua; Zhou, Jialiang; Lu, Kai; Li, Keqing; Zhou, Qiang

    2015-11-01

    When cutworm Spodoptera litura larvae were fed on the diets with different lead (Pb) concentrations for one or five generations, changes in growth and food utilization were recorded; Pb accumulations were detected by Atomic Absorption Spectrophotometer; changes in midgut cell ultrastructure were observed by Transmission Electron Microscopy (TEM). The effects of Pb stress on S. litura growth and food utilization differed significantly between insects of the 1st and 5th generation. The male-female rate of 200mgkg(-1) Pb treatment from the 1st generation and 50mgkg(-1) Pb treatment from the 5th generation was significantly higher than control. No significant difference of Pb accumulations was found in larvae, pupae and adults between the 1st and 5th generation. No significant difference of Pb accumulations in corresponding tissues of larvae was found between male and female. Compared to fat body, hemolymph, head, foregut and hindgut, the highest Pb accumulation was found in migut of larvae exposed to 200mgkg(-1) Pb. TEM showed that expanded intercellular spaces were observed in Pb-treated midgut cells. The nuclei were strongly destroyed by Pb stress, evidenced by chromatin condensation and destroyed nuclear envelope. Mitochondria became swollen with some broken cristae after exposure to Pb. Therefore, neither gender nor progeny difference was present in Pb accumulations of S. litura, although effects of Pb stress on S. litura growth and food utilization differed from different generations and genders. Pb accumulations in midgut caused pathological changes in cells ultrastructure, possibly reflected the growth and food utilization of S. litura. PMID:26248226

  10. Modulation of reproductive behaviors by non-host volatiles in the polyphagous Egyptian cotton leafworm, Spodoptera littoralis.

    PubMed

    Binyameen, Muhammad; Hussain, Altaf; Yousefi, Fatemeh; Birgersson, Göran; Schlyter, Fredrik

    2013-10-01

    In order to locate mates, food, and oviposition sites, insects mainly rely on volatile cues released by their sexual partners, food sources, and host and non-host plants. Calling, mating, and oviposition behaviors, as well as fecundity and longevity, of newly emerged Spodoptera littoralis (Bois.) moths were recorded in the presence of volatiles from leaves of a host plant, Gossypium hirsutum (cotton) and two non-host plants, Adhatoda vasica (Av) or Picea abies (spruce), either alone or in host/non-host combinations. Females exposed to cotton volatiles started calling earlier than females exposed to non-host plant volatiles (NHV), or the blank control. Likewise, moth pairs exposed to cotton volatiles started mating earlier than the other treatments. The period of calling in females alone was longer than females kept with males, having the opportunity to mate. However, the callings, as well as mating durations in the moth pairs, in different treatments were not different. Longevity was decreased either in the absence of cotton or the presence of Av, and spruce leaves. Fecundity was reduced in moths exposed to a combination of spruce and cotton. The effect of NHV on attraction of 2-3-day-old male moths towards a pheromone (Ph) source was studied in a wind tunnel. In the no-choice assay, more males arrived at close approach and landed on the Ph source when the host plant, cotton, was offered in the background as compared to the non-hosts. In the dual-choice assay, more males landed on the Ph source in front of the host plant compared to the Ph source in front of non-hosts. Gas chromatography-electroantennographic detection on female S. littoralis revealed five antennally active compounds in headspace collections of spruce and three compounds in Av. PMID:24105603

  11. Characterization of DNA Topoisomerase-1 in Spodoptera exigua for Toxicity Evaluation of Camptothecin and Hydoxy-Camptothecin

    PubMed Central

    Zhang, Yanning; He, Weizhi; Yang, Jingjing; Li, Chuanren; Jiang, Hongyun

    2013-01-01

    Camptothecin (CPT), a plant alkaloid originally isolated from the native Chinese tree, Camptotheca acuminate, exerts the toxic effect by targeting eukaryotic DNA topoisomerase 1 (DNA Topo1). Besides as potent anti-cancer agents, CPT and its derivatives are now being explored as potential pesticides for insect control. In this study, we assessed their toxicity to an insect homolog, the Topo1 protein from beet armyworms (Spodoptera exigua Hübner), a worldwide pest of many important crops. The S. exigua Topo1 gene contains an ORF of 2790 base pairs that is predicted to encode a polypeptide of 930 amino acids. The deduced polypeptide exhibits polymorphism at residue sites V420, L530, A653 and T729 (numbered according to human Topo1) among insect species, which are predicted to confer sensitivity to CPT. The DNA relaxation activity of this protein was subsequently examined using a truncated form that contained the residues 337–930 and was expressed in bacteria BL21 cells. The purified protein retained the ability to relax double-stranded DNA and was susceptible to CPT and its derivative hydroxy-camptothecin (HCPT) in a dose-dependent manner. The same inhibitory effect was also found on the native Topo1 extracted from IOZCAS-Spex-II cells, a cell line established from beet armyworms. Additionally, CPT and HCPT treatment reduced the steady accumulation of Topo1 protein despite the increased mRNA expression in response to the treatment. Our studies provide information of the S. exigua Topo1 gene and its amino acid polymorphism in insects and uncover some clues about potential mechanisms of CPT toxicity against insect pests. These results also are useful for development of more effective Topo1-targeted CPT insecticides in the future. PMID:23451051

  12. PGE(2) induces oenocytoid cell lysis via a G protein-coupled receptor in the beet armyworm, Spodoptera exigua.

    PubMed

    Shrestha, Sony; Stanley, David; Kim, Yonggyun

    2011-11-01

    Eicosanoids mediate cellular and humoral immune responses in the beet armyworm, Spodoptera exigua, including activation of prophenoloxidase (PPO). PPO activation begins with release of its inactive zymogen, PPO, from oenocytoids in response to prostaglandins (PGs). Based on the biomedical literature, we hypothesized that PGs exert their actions via specific G protein-coupled receptor(s) in S. exigua. This study reports a G protein-coupled receptor (Se-hcPGGPCR1) gene, which is expressed in the hemocytes of S. exigua. The Se-hcPGGPCR1 consists of 420 amino acids and belongs to rhodopsin-type GPCRs. The high content of hydrophobic amino acid residues within the Se-hcPGGPCR1 protein is explained by prediction of seven-transmembrane domains that are characteristic of these GPCRs. Except for the eggs, Se-hcPGGPCR1 was expressed in all life stages. During the larval stage, it was expressed in hemocytes and gut, but not in fat body nor in epidermis. Real time quantitative RT-PCR showed that bacterial challenge induced more than 20-fold increases in its expression level. Fluorescence in situ hybridization showed that Se-hcPGGPCR1 was expressed in a specific hemocyte type, the oenocytoids. A specific eicosanoid, PGE(2), significantly induced oenocytoid lysis and increased PO activity in the plasma. In contrast, when Se-hcPGGPCR1 expression was suppressed by RNA interference (RNAi), the oenocytoid lysis and the PO activation in response to PGE(2) were not elevated above basal levels. A binding assay using intracellular calcium mobilization showed that the RNAi-treated hemocytes were significantly less responsive to PGE(2) than the control hemocytes. These results support our hypothesis with the specific finding that PGE(2) acts through Se-hcPGGPCR1 to activate PPO by lysing oenocytoids. PMID:21867708

  13. Biopotency of serine protease inhibitors from cowpea (Vigna unguiculata) seeds on digestive proteases and the development of Spodoptera littoralis (Boisduval).

    PubMed

    Abd El-latif, Ashraf Oukasha

    2015-05-01

    Serine protease inhibitors (PIs) have been described in many plant species and are universal throughout the plant kingdom, where trypsin inhibitors is the most common type. In the present study, trypsin and chymotrypsin inhibitory activity was detected in the seed flour extracts of 13 selected cultivars/accessions of cowpea. Two cowpea cultivars, Cream7 and Buff, were found to have higher trypsin and chymotrypsin inhibitory potential compared to other tested cultivars for which they have been selected for further purification studies using ammonium sulfate fractionation and DEAE-Sephadex A-25 column. Cream7-purified proteins showed two bands on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) corresponding to molecular mass of 17.10 and 14.90 kDa, while the purified protein from Buff cultivar showed a single band corresponding mass of 16.50 kDa. The purified inhibitors were stable at temperature below 60°C and were active at wide range of pH from 2 to 12. The kinetic analysis revealed noncompetitive type of inhibition for both inhibitors against both enzymes. The inhibitor constant (Ki ) values suggested high affinity between inhibitors and enzymes. Purified inhibitors were found to have deep and negative effects on the mean larval weight, larval mortality, pupation, and mean pupal weight of Spodoptera littoralis, where Buff PI was more effective than Cream7 PI. It may be concluded that cowpea PI gene(s) could be potential insect control protein for future studies in developing insect-resistant transgenic plants. PMID:25524889

  14. The role of cytochrome c on apoptosis induced by Anagrapha falcifera multiple nuclear polyhedrosis virus in insect Spodoptera litura cells.

    PubMed

    Liu, Kaiyu; Shu, Duanyang; Song, Na; Gai, Zhongchao; Yuan, Yuan; Li, Juan; Li, Min; Guo, Shuying; Peng, Jianxin; Hong, Huazhu

    2012-01-01

    There are conflicting reports on the role of cytochrome c during insect apoptosis. Our previous studies have showed that cytochrome c released from the mitochondria was an early event by western blot analysis and caspase-3 activation was closely related to cytochrome c release during apoptosis induced by baculovirus in Spodoptera litura cells (Sl-1 cell line). In the present study, alteration in mitochondrial morphology was observed by transmission electron microscopy, and cytochrome c release from mitochondria in apoptotic Sl-1 cells induced with Anagrapha falcifera multiple nuclear polyhedrosis virus (AfMNPV) has further been confirmed by immunofluoresence staining protocol, suggesting that structural disruption of mitochondria and the release of cytochrome c are important events during Lepidoptera insect cell apoptosis. We also used Sl-1 cell-free extract system and the technique of RNA interference to further investigate the role of cytochrome c in apoptotic Sl-1 cells induced by AfMNPV. Caspase-3 activity in cell-free extracts supplemented with exogenous cytochrome c was determined and showed an increase with the extension of incubation time. DsRNA-mediated silencing of cytochrome c resulted in the inhibition of apoptosis and protected the cells from AfMNPV-induced cell death. Silencing of expression of cytochrome c had a remarkable effect on pro-caspase-3 and pro-caspase-9 activation and resulted in the reduction of caspase-3 and caspase-9 activity in Sl-1 cells undergoing apoptosis. Caspase-9 inhibitor could inhibit activation of pro-caspase-3, and the inhibition of the function of Apaf-1 with FSBA blocked apoptosis, hinting that Apaf-1 could be involved in Sl-1 cell apoptosis induced by AfMNPV. Taken together, these results strongly demonstrate that cytochrome c plays an important role in apoptotic signaling pathways in Lepidopteran insect cells. PMID:22952575

  15. Dynamics of Membrane Potential Variation and Gene Expression Induced by Spodoptera littoralis, Myzus persicae, and Pseudomonas syringae in Arabidopsis

    PubMed Central

    Bricchi, Irene; Bertea, Cinzia M.; Occhipinti, Andrea; Paponov, Ivan A.; Maffei, Massimo E.

    2012-01-01

    Background Biotic stress induced by various herbivores and pathogens invokes plant responses involving different defense mechanisms. However, we do not know whether different biotic stresses share a common response or which signaling pathways are involved in responses to different biotic stresses. We investigated the common and specific responses of Arabidopsis thaliana to three biotic stress agents: Spodoptera littoralis, Myzus persicae, and the pathogen Pseudomonas syringae. Methodology/Principal Findings We used electrophysiology to determine the plasma membrane potential (Vm) and we performed a gene microarray transcriptome analysis on Arabidopsis upon either herbivory or bacterial infection. Vm depolarization was induced by insect attack; however, the response was much more rapid to S. littoralis (30 min −2 h) than to M. persicae (4–6 h). M. persicae differentially regulated almost 10-fold more genes than by S. littoralis with an opposite regulation. M. persicae modulated genes involved in flavonoid, fatty acid, hormone, drug transport and chitin metabolism. S. littoralis regulated responses to heat, transcription and ion transport. The latest Vm depolarization (16 h) was found for P. syringae. The pathogen regulated responses to salicylate, jasmonate and to microorganisms. Despite this late response, the number of genes differentially regulated by P. syringae was closer to those regulated by S. littoralis than by M. persicae. Conclusions/Significance Arabidopsis plasma membranes respond with a Vm depolarization at times depending on the nature of biotic attack which allow setting a time point for comparative genome-wide analysis. A clear relationship between Vm depolarization and gene expression was found. At Vm depolarization timing, M. persicae regulates a wider array of Arabidopsis genes with a clear and distinct regulation than S. littoralis. An almost completely opposite regulation was observed between the aphid and the pathogen, with the former

  16. Bacillus thuringiensis Cry1Ca-resistant Spodoptera exigua lacks expression of one of four Aminopeptidase N genes

    PubMed Central

    Herrero, Salvador; Gechev, Tsanko; Bakker, Petra L; Moar, William J; de Maagd, Ruud A

    2005-01-01

    Background Insecticidal toxins from Bacillus thuringiensis bind to receptors on midgut epithelial cells of susceptible insect larvae. Aminopeptidases N (APNs) from several insect species have been shown to be putative receptors for these toxins. Here we report the cloning and expression analysis of four APN cDNAs from Spodoptera exigua. Results Suppression Subtractive Hybridization (SSH) was used to construct cDNA libraries of genes that are up-and down-regulated in the midgut of last instar larvae of beet armyworm, S. exigua exposed to B. thuringiensis Cry1Ca toxin. Among the clones from the SSH libraries, cDNA fragments coding for two different APNs were obtained (APN2 and APN4). A similar procedure was employed to compare mRNA differences between susceptible and Cry1Ca resistant S. exigua. Among the clones from this last comparison, cDNA fragments belonging to a third APN (APN1) were detected. Using sequences obtained from the three APN cDNA fragments and degenerate primers for a fourth APN (APN3), the full length sequences of four S. exigua APN cDNAs were obtained. Northern blot analysis of expression of the four APNs showed complete absence of APN1 expression in the resistant insects, while the other three APNs showed similar expression levels in the resistant and susceptible insects. Conclusion We have cloned and characterized four different midgut APN cDNAs from S. exigua. Expression analysis revealed the lack of expression of one of these APNs in the larvae of a Cry1Ca-resistant colony. Combined with previous evidence that shows the importance of APN in the mode of action of B. thuringiensis toxins, these results suggest that the lack of APN1 expression plays a role in the resistance to Cry1Ca in this S. exigua colony. PMID:15978131

  17. Identification of a novel cytochrome P450 CYP321B1 gene from tobacco cutworm moth (Spodoptera litura) and RNA interference to evaluate its role in commonly used insecticides.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    BACKGROUND: Insect cytochrome P450 monooxygenases (CYPs or P450s) play an important role in detoxifying insecticides leading to resistance in insect populations. A polyphagous pest, Spodoptera litura (Lepidoptera, Noctuidae) has been shown to be resistant to a wide range of insecticides. In this stu...

  18. Does secondary plant metabolism provide a mechanism for plant defenses in the tropical soda apple Solanum viarum (Solanales: Solanaceae) against the beet armyworm Spodoptera exigua and southern armyworm S. eridania?

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Survival assays were conducted with beet armyworm Spodoptera exigua and southern armyworm S. eridania with tropical soda apple Solanum viarum a relative of tomato. In addition, polyphenol oxidase (PPO) enzyme assays were conducted to determine if secondary plant defense compounds are being produce...

  19. Characterization of an Egyptian Spodoptera littoralis nucleopolyhedrovirus and a possible use of a highly conserved region from polyhedrin gene for nucleopolyhedrovirus detection

    PubMed Central

    Seufi, AlaaEddeen M

    2008-01-01

    An Egyptian isolate of Spodoptera littoralis nucleopolyhedrovirus (SpliNPV) was tested for its potential as biocontrol agent in comparison to Autographa californica multiple nucleopolyhedrovirus (AcMNPV). Comparative assays of SpliNPV and AcMNPV against 2nd instar larvae of Spodoptera littoralis revealed 4-fold greater susceptibility of S. littoralis to AcMNPV than to SpliNPV based on LC50 values for the two viruses. The LT50s determined for SpliNPV and AcMNPV using LC50 of the virus against 2nd instar larvae were 4.2 and 5.8 days, respectively. A DNA segment of 405 bp containing highly conserved region from polyhedrin gene of SpliNPV (Polh-cr) was successfully amplified by PCR. Subsequently, this DNA segment was cloned and sequenced. Nucleotide sequence and its deduced amino acid sequence were compared to all available sequences in GenBank. Sequence alignment results revealed that Polh-cr showed significant similarities with 91 different baculovirus isolates. The percentage of homology ranged from 78% for Plusia orichalcea NPV to 99% for SpliNPV. This highly conserved region provides a candidate that could be used in easy, fast and economic prospective systems for virus detection as well as in biological control strategies. PMID:18215282

  20. Temperature- and CO2-dependent life table parameters of Spodoptera litura (Noctuidae: Lepidoptera) on sunflower and prediction of pest scenarios.

    PubMed

    Manimanjari, D; Srinivasa Rao, M; Swathi, P; Rama Rao, C A; Vanaja, M; Maheswari, M

    2014-01-01

    Predicted increase in temperature and atmospheric CO2 concentration will influence the growth of crop plants and phytophagous insects. The present study, conducted at the Central Research Institute for Dryland Agriculture, Hyderabad, India, aimed at (1) construction of life tables at six constant temperatures viz., 20, 25, 27, 30, 33, and 35 ± 0.5 °C for Spodoptera litura (Fabricius) (Noctuidae: Lepidoptera) reared on sunflower (Helianthus annus L.) grown under ambient and elevated CO2 (eCO2) (550 ppm) concentration in open top chambers and (2) prediction of the pest status in near future (NF) and distant future (DF) climate change scenarios at major sunflower growing locations of India. Significantly lower leaf nitrogen, higher carbon and higher relative proportion of carbon to nitrogen (C:N) were observed in sunflower foliage grown under eCO2 over ambient. Feeding trials conducted on sunflower foliage obtained from two CO2 conditions showed that the developmental time of S. litura (Egg to adult) declined with increase in temperature and was more evident at eCO2. Finite (λ) and intrinsic rates of increase (r(m)), net reproductive rate (Ro), mean generation time, (T) and doubling time (DT) of S. litura increased significantly with temperature up to 27-30 °C and declined with further increase in temperature. Reduction of 'T' was observed from maximum value of 58 d at 20 °C to minimum of 24.9 d at 35 °C. The DT of population was higher (5.88 d) at 20 °C and lower (3.05 d) at 30 °C temperature of eCO2. The data on these life table parameters were plotted against temperature and two nonlinear models were developed separately for each of the CO2 conditions for predicting the pest scenarios. The NF and DF scenarios temperature data of four sunflower growing locations in India is based on PRECIS A1B emission scenario. It was predicted that increased 'rm', 'λ', and 'Ro' and reduced 'T' would occur during NF and DF scenario over present period at all

  1. COI1-Regulated Hydroxylation of Jasmonoyl-L-isoleucine Impairs Nicotiana attenuata's Resistance to the Generalist Herbivore Spodoptera litura.

    PubMed

    Luo, Ji; Wei, Kun; Wang, Shuanghua; Zhao, Weiye; Ma, Canrong; Hettenhausen, Christian; Wu, Jinsong; Cao, Guoyan; Sun, Guiling; Baldwin, Ian T; Wu, Jianqiang; Wang, Lei

    2016-04-13

    The phytohormone jasmonoyl-L-isoleucine (JA-Ile) is well-known as the key signaling molecule that elicits plant defense responses after insect herbivory. Oxidation, which is catalyzed by the cytochrome P450s of the CYP94 family, is thought to be one of the main catabolic pathways of JA-Ile. In this study, we identified four CYP94B3 homologues in the wild tobacco plant Nicotiana attenuata. Individually silencing the four homologues revealed that NaCYP94B3 like-1 and NaCYP94B3 like-2, but not NaCYP94B3 like-3 and NaCYP94B3 like-4, are involved in the C-12-hydroxylation of JA-Ile. Simultaneously silencing three of the NaCYP94B3 like genes, NaCYP94B3 like-1, -2, and -4, in the VIGS-NaCYP94B3s plants doubled herbivory-induced JA-Ile levels and greatly enhanced plant resistance to the generalist insect herbivore, Spodoptera litura. The poor larval performance was strongly correlated with the high concentrations of several JA-Ile-dependent direct defense metabolites in VIGS-NaCYP94B3s plants. Furthermore, we show that the abundance of 12-hydroxy-JA-Ile was dependent on JA-Ile levels as well as COI1, the receptor of JA-Ile. COI1 appeared to transcriptionally control NaCYP94B3 like-1 and -2 and thus regulates the catabolism of its own ligand molecule, JA-Ile. These results highlight the important role of JA-Ile degradation in jasmonate homeostasis and provide new insight into the feedback regulation of JA-Ile catabolism. Given that silencing these CYP94 genes did not detectably alter plant growth and highly increased plant defense levels, we propose that CYP94B3 genes can be potential targets for genetic improvement of herbivore-resistant crops. PMID:26985773

  2. Chlorantraniliprole/lambda-cyhalothrin, a new insecticide mixture to control Tuta absoluta and Spodoptera littoralis in tomato.

    PubMed

    Fanigliulo, A; Mancino, O; Fanti, P; Crescenzi, A

    2012-01-01

    A study was performed in the period May-July 2011 by Bioagritest test facility according to EPPO guidelines and Principles of Good Experimental Practice (GEP), in the land of Eboli (SA), southern Italy, with the purpose to test a new insecticide mixture in the defense strategies of processing tomato against Tuta absoluta and Spodoptera littoralis. The insecticide mixture Chlorantraniliprole 100 g/lambda-Cyhalothrin 50 g/l 0,4 It/ha was applied in a tomato field in four (A-B-C-D) application timings (a.t.) alone (treatment 2) and in two a.t. (A-B) combined with emamectin benzoate 0.5%, 1,5 Kg/ha (a.t. C-D) (treatment 3). Its effect in containing insect populations was compared with that exerted by emamectin benzoate 0.5%, 1,5 Kg/ha (a.t. A-B-C-D) (treatment 4) and the combination of chlorantraniliprole 35%, 0.115 Kg/ha (a.t. A-B) and indoxacarb 30% 0.125 Kg/ha (a.t. C-D) (treatment 5). Application of treatments (trt.) and mode of assessment (2 assessments), recording and measurements followed the guidelines foreseen by EPPO Standards PP1/150 and PP1/275. The lepidoptera infestation in the tomato crop was very high on both the leaves and fruits, and this enabled us to evaluate the efficacy of the tested products in conditions of severe pests infestation. If we consider the leaf damage, either as number of mines by T. absoluta or as % of leaf erosion by S. litteralis, all the treatments values were statistically lower than control, but with no difference among them. But when using as an infestation threshold the percentage of damaged fruits (much more relevant in economic terms), we observed statistically significant differences among treatments. Actually, treatment with chlorantraniliprole-lambda-cyhalothrin mixture was significantly more effective than the one with emamectin benzoate in reducing the attack of T. absoluta on tomato fruits, in both dates of assessments. As concerns damage by S. littoralis on the fruits, at the first assessment, trt. 2 and trt. 5

  3. Effects of some botanical extracts on the midgut, integument and fat body of the cotton leaf worm, Spodoptera littoralis (Lepidoptera: Noctuidae).

    PubMed

    Khatter, Najat A

    2010-08-01

    Botanical extracts (8%) of four plants (Artemisia monosperma, Zygophyllum cocccineum, Lupinus termis and Brassica tournifortii) fed to the 4th larval instars of Spodoptera littoralis induced histopathological changes in the structure of the midgut, integument and fat body of the 5th instars. Zygophyllum cocci-neum and Lupinus termis induced severe damages in the midgut. The integument of treated larvae showed degeneration in the cuticle and epidermal cells which were also detached from each other. Water extracts of A. monosperma, Z. coccinieum and L. termis were the most promising in inducing shrinkage in the fat body cells and detachment of midgut muscle layers. Also, the degeneration of the midgut membrane and epithelial layer occurs in different degrees with the tested plants. This study supports the use of botanical extracts in pest control programs of lepidopterous insects. PMID:21246948

  4. Biological activities of Solanum pseudocapsicum (Solanaceae) against cotton bollworm, Helicoverpa armigera Hübner and armyworm, Spodoptera litura Fabricius (Lepidotera: Noctuidae)

    PubMed Central

    Jeyasankar, Alagarmalai; Premalatha, Selvaraj; Elumalai, Kuppusamy

    2012-01-01

    Objective To evaluate the antifeedant, insecticidal and growth inhibition activities of Solanum pseudocapsicum (S. pseudocapsicum) seed extracts against Spodoptera litura (S. litura) and Helicoverpa armigera (H. armigera). Methods Hexane, diethyl ether, dichloromethane and ethyl acetate seed extracts were prepared and tested for antifeedant, insecticidal and growth inhibitory activities against fourth instar larvae of S. litura and H. armigera. Results Ethyl acetate extract showed promising antifeedant and insecticidal activities against S. litura and H. armigera. Percentage of deformed larvae, pupae and adults were maximum in treatment of ethyl acetate extract. Percentage of successful adult emergence was deteriorated by seeds on extract treated larvae. Conclusions Ethyl acetate extracts of S. pseudocapsicum, showed higher efficiency of antifeedant, insecticidal and growth inhibition activities. Hence, it can be used to controll agricultural insect pests, S. litura and H. armigera. PMID:23593579

  5. Hyperactivity and tree-top disease induced by the baculovirus AcMNPV in Spodoptera exigua larvae are governed by independent mechanisms

    NASA Astrophysics Data System (ADS)

    van Houte, Stineke; Ros, Vera I. D.; van Oers, Monique M.

    2014-04-01

    Although many parasites are known to manipulate the behavior of their hosts, the mechanisms underlying such manipulations are largely unknown. Baculoviruses manipulate the behavior of caterpillar hosts by inducing hyperactivity and by inducing climbing behavior leading to death at elevated positions (tree-top disease or Wipfelkrankheit). Whether hyperactivity and tree-top disease are independent manipulative strategies of the virus is unclear. Recently, we demonstrated the involvement of the protein tyrosine phosphatase ( ptp) gene of the baculovirus Autographa californica multiple nucleopolyhedrovirus (AcMNPV) in the induction of hyperactivity in Spodoptera exigua larvae. Here we show that AcMNPV ptp is not required for tree-top disease, indicating that in S. exigua baculovirus-induced hyperactivity and tree-top disease are independently induced behaviors that are governed by distinct mechanisms.

  6. Characterization of Multisugar-Binding C-Type Lectin (SpliLec) from a Bacterial-Challenged Cotton Leafworm, Spodoptera littoralis

    PubMed Central

    Seufi, AlaaEddeen M.; Galal, Fatma H.; Hafez, Elsayed E.

    2012-01-01

    Background Various proteins that display carbohydrate-binding activity in a Ca2+-dependent manner are classified into the C-type lectin family. They have one or two C-type carbohydrate-recognition domains (CRDs) composed of 110–130 amino acid residues in common. C-type lectins mediate cell adhesion, non-self recognition, and immuno-protection processes in immune responses and thus play significant roles in clearance of invaders, either as cell surface receptors for microbial carbohydrates or as soluble proteins existing in tissue fluids. The lectin of Spodoptera littoralis is still uncharacterized. Methodology A single orf encoding a deduced polypeptide consisting of an 18-residue signal peptide and a 291-residue mature peptide, termed SpliLec, was isolated from the haemolymph of the cotton leafworm, S. littoralis, after bacterial challenge using RACE-PCR. Sequence analyses of the data revealed that SpliLec consists of two CRDs. Short-form CRD1 and long-form CRD2 are stabilized by two and three highly conserved disulfide bonds, respectively. SpliLec shares homology with some dipteran lectins suggesting possible common ancestor. The purified SpliLec exhibited a 140-kDa molecular mass with a subunit molecular mass of 35 kDa. The hemagglutination assays of the SpliLec confirmed a thermally stable, multisugar-binding C-type lectin that binds different erythrocytes. The purified SpliLec agglutinated microorganisms and exhibited comparable antimicrobial activity against gram (+) and gram (−) bacteria too. Conclusions Our results suggested an important role of the SpliLec gene in cell adhesion and non-self recognition. It may cooperate with other AMPs in clearance of invaders of Spodoptera littoralis. PMID:22916161

  7. Cloning, Expression Analysis, and Molecular Modeling of the Gamma-Aminobutyric Acid Receptor Alpha2 Subunit Gene from the Common Cutworm, Spodoptera litura

    PubMed Central

    Zuo, Hongliang; Gao, Lu; Hu, Zhen; Liu, Haiyuan; Zhong, Guohua

    2013-01-01

    Intensive research on the molecule structures of the gamma-nminobutyric acid (GABA) receptor in agricultural pests has great significance to the mechanism investigation, resistance prevention, and molecular design of novel pesticides. The GABA receptor a2 (SlGABARα2) subunit gene in Spodoptera litura (Fabricius) (Lepidoptera: Noctuidae) was cloned using the technologies of reverse transcription PCR and rapid amplification of cDNA ends. The gemonic DNA sequence of SlGABARα2 has 5164 bp with 8 exons and 7 introns that were in accordance with the GT-AG splicing formula. The complete mRNA sequence of SlGABARα2 was 1965 bp, with an open reading frame of 1500 bp encoding a protein of 499 amino acids. The GABA receptor is highly conserved among insects. The conserved regions include several N-glycosylation, Oglycosylation, and phosphorylation sites, as well as 4 transmembrane domains. The identities that SlGABARα2 shared with the GABA receptor a2 subunit of Spodoptera exigua, Heliothis virescens, Chilo suppressalis, Plutella xylostella, Bombyx mori ranged from 99.2% to 87.2% at the amino acid level. The comparative 3-dimensional model of SlGABARα2 showed that its tertiary structure was composed of 4 major α-helixes located at the 4 putative transmembrane domains on one side, with some β-sheets and 1 small α-helix on the other side. SlGABARα2 may be attached to the membrane by 4 α-helixes that bind ions in other conserved domains to transport them through the membrane. The results of quantitative real time PCR demonstrated that SlGABARα2 was expressed in all developmental stages of S. litura. The relative expression level of SlGABARα2 was the lowest in eggs and increased with larval growth, while it declined slightly in pupae and reached the peak in adults. The expressions of SlGABARα2 in larvae varied among different tissues; it was extremely high in the brain but was low in the midgut, epicuticle, Malpighian tube, and fat body. PMID:23909412

  8. Cloning, expression analysis, and molecular modeling of the gamma-aminobutyric acid receptor alpha2 subunit gene from the common cutworm, Spodoptera litura.

    PubMed

    Zuo, Hongliang; Gao, Lu; Hu, Zhen; Liu, Haiyuan; Zhong, Guohua

    2013-01-01

    Intensive research on the molecule structures of the gamma-nminobutyric acid (GABA) receptor in agricultural pests has great significance to the mechanism investigation, resistance prevention, and molecular design of novel pesticides. The GABA receptor a2 (SlGABARα2) subunit gene in Spodoptera litura (Fabricius) (Lepidoptera: Noctuidae) was cloned using the technologies of reverse transcription PCR and rapid amplification of cDNA ends. The gemonic DNA sequence of SlGABARα2 has 5164 bp with 8 exons and 7 introns that were in accordance with the GT-AG splicing formula. The complete mRNA sequence of SlGABARα2 was 1965 bp, with an open reading frame of 1500 bp encoding a protein of 499 amino acids. The GABA receptor is highly conserved among insects. The conserved regions include several N-glycosylation, Oglycosylation, and phosphorylation sites, as well as 4 transmembrane domains. The identities that SlGABARα2 shared with the GABA receptor a2 subunit of Spodoptera exigua, Heliothis virescens, Chilo suppressalis, Plutella xylostella, Bombyx mori ranged from 99.2% to 87.2% at the amino acid level. The comparative 3-dimensional model of SlGABARα2 showed that its tertiary structure was composed of 4 major α-helixes located at the 4 putative transmembrane domains on one side, with some β-sheets and 1 small α-helix on the other side. SlGABARα2 may be attached to the membrane by 4 α-helixes that bind ions in other conserved domains to transport them through the membrane. The results of quantitative real time PCR demonstrated that SlGABARα2 was expressed in all developmental stages of S. litura. The relative expression level of SlGABARα2 was the lowest in eggs and increased with larval growth, while it declined slightly in pupae and reached the peak in adults. The expressions of SlGABARα2 in larvae varied among different tissues; it was extremely high in the brain but was low in the midgut, epicuticle, Malpighian tube, and fat body. PMID:23909412

  9. Regulation of hemolymph trehalose level by an insulin-like peptide through diel feeding rhythm of the beet armyworm, Spodoptera exigua.

    PubMed

    Kim, Yonggyun; Hong, Youkyeong

    2015-06-01

    Like vertebrate insulins, some insect insulin-like peptides (ILPs) play crucial roles in controlling immature growth, adult lifespan, and hemolymph sugar level. An ILP gene (SeILP1) was predicted from a transcriptome database of Spodoptera exigua. SeILP1 encodes 95 amino acid sequence and shares sequence homologies (33-83%) with other insect ILPs, in which six conserved cysteine residues are found in the predicted B-A chains. SeILP1 was expressed in all developmental stages of S. exigua. However, SeILP1 expression was tissue-specific because the transcript was detected in fat body and epidermis, but not in hemocytes and gut. Its expression increased with feeding activity. Hemolymph trehalose levels of the fifth instar larvae maintained a relatively constant level at 2.31±0.62mM. However, starvation induced a significant increase of the hemolymph trehalose level by more than twofold in 48h, at which few SeILP1 was transcribed. RNA interference of SeILP1 using its specific double-stranded RNA induced a significant increase of hemolymph trehalose level. Interestingly, a bovine insulin decreased hemolymph trehalose level in a dose-dependent manner. These results indicate that SeILP1 plays a role in suppressing hemolymph trehalose level in S. exigua. PMID:25703302

  10. Insecticidal Effect of Chrysanthemum coronarium L. Flowers on the Pest Spodoptera littoralis Boisd and its Parasitoid Microplitis rufiventris Kok. with Identifying the Chemical Composition

    NASA Astrophysics Data System (ADS)

    Shonouda, Mourad L.; Osman, Salah; Salama, Osama; Ayoub, Amal

    The flower extract of Chrysanthemum coronarium L. and their fractions have shown insecticidal effect on the cotton leaf worm Spodoptera littoralis. The third instar larvae fed for two days on treated leaves were more susceptible to plant extracts and to their ethyl acetate and chloroform fractions. The active lowest concentration (5%) of the flower fractions showed no significant effect on the percent reduction of emerged adult parasitoids, Microplitis rufiventris Kok. GC/MS analysis revealed that the major constituents in ethyl acetate fraction were 3-dihydro-methylene-2- (3H) furanone (17.8%), jasmolin I (15.6%), carveol 1 (13.6%), phosphoric acid, tributyl ester (11.4%) and cinerin II (11.1%), while those of chloroform fraction were 5-hydroxy-3 methyl-1H-pyrazole (42.7%) and carveol 1(24.8%). The medicinal plant C. coronarium seems to be a promising plant for application in integrated pest management due to its safety to the surrounding environment.

  11. Imperfection works: Survival, transmission and persistence in the system of Heliothis virescens ascovirus 3h (HvAV-3h), Microplitis similis and Spodoptera exigua

    PubMed Central

    Li, Shun-Ji; Hopkins, Richard J.; Zhao, Yi-Pei; Zhang, Yun-Xuan; Hu, Jue; Chen, Xu-Yang; Xu, Zhi; Huang, Guo-Hua

    2016-01-01

    Ascoviruses are insect-specific large DNA viruses that mainly infect noctuid larvae, and are transmitted by parasitoids in the fields. Heliothis virescens ascovirus 3h (HvAV-3h) has been recently isolated from Spodoptera exigua, without parasitoid vector identified previously. Here we report that Microplitis similis, a solitary endoparasitoid wasp, could transmit HvAV-3h between S. exigua larvae in the laboratory. When the female parasitoid wasp acquired the virus and served as a vector, the period of virion viability on the ovipositor was 4.1 ± 1.4 days. Infected host larvae were still acceptable for egg laying by parasitoids, and the parasitoids thereafter transmitted virus to healthy hosts. Virus acquisition occurred only from donor hosts between 3 and 9 days post infection. The peak of virus acquisition (80.9 ± 6.3%) was found when M. similis wasps oviposited in larvae that had been inoculated with the virus 7 days previously. When virus infection of the host took place during the life cycle of the parasitoid wasp, it caused 1- to 4-day-old immature parasitoids death in the host, whilst a small proportion of 5- to 6-day-old and the majority of 7-day-old parasitoids larvae survived from the virus-infected hosts. Viral contamination did not reduce the life span or fecundity of female M. similis. PMID:26878829

  12. A new cell line from the fat body of Spodoptera litura (Lepidoptera, Noctuidae) and detection of lysozyme activity release upon immune stimulation.

    PubMed

    Tateishi, Ken; Kasahara, Yuichi; Watanabe, Kazuyo; Hosokawa, Nobuo; Doi, Hiroyasu; Nakajima, Kaori; Adachi, Hayamitsu; Nomoto, Akio

    2015-01-01

    A new cell line, designated NIAS-SL64, was established from the fat body of the fifth instar larvae of the common cutworm Spodoptera litura. NIAS-SL64 cells grew as spindle-shaped and non-adherent cells in the insect-specific cell culture medium MGM-450 supplemented with 10% fetal bovine serum. Criterions for the establishment of the NIAS-SL64 cell line is spindle shape and length (30~90 μm) stabilized after 100 passages. The doubling time of the cells was 24 h at 25°C. Lipopolysaccharide significantly stimulated the release of lysozyme activity by NIAS-SL64 cells. Lysozyme is one of the components of the innate immunity and plays important role as lytic enzyme in infection. Lysozyme activity released from NIAS-SL64 would be a marker for immune response. The released lysozyme activity critically depends on morphology of the cells and would be a criterion of the establishment of the cell line. Lysozyme activity was suppressed in a dose-dependent manner by the immunosuppressive agent cyclosporin A. PMID:25172011

  13. The Homologous Regions of Spodoptera litura (Lepidoptera: Noctuidae) Nucleopolyhedrovirus II Have Both the Function as Origin of DNA Replication and Enhancer

    PubMed Central

    Liu, Huifen; Gao, Huiju; Guo, Guang; Li, Yunzhi; Li, Yinü; Wang, Jinhui; Zhang, Zhifang; Yu, Zhencheng

    2015-01-01

    In the genome sequence of the Spodoptera litura nucleopolyhedrovirus II (SpltNPVII), seven homologous regions (hrs), Sphr1-7, were identified. Each of them composed of three to eight 64-bp highly conserved sequences, and each contained a 24-bp imperfect palindrome. A transient expression assay demonstrated that the expression of SpltNPVII-ie1 promoter-driven luciferase gene was enhanced between 3- and 13-fold by infection of SpltNPVII in Spli221 cells. Real-time polymerase chain reaction confirmed each of seven hrs could function as origin (ori) of viral DNA replication. This suggests that these hrs are bifunctional, having both ori and enhancer activities for transcription. In addition, the potential of seven hrs as origins had a significantly positive correlation with the number of their palindromes (r = 0.847, Sig: 0.016 < 0.05), and enhancer efficiency had a significantly positive correlation with the number of characteristic motifs (r = 0.893, Sig: 0.007 < 0.01). The efficiency of replication and enhancement of each hr both increased with increasing total numbers of palindromes, repeat sequences, and characteristic motifs. In addition, a single 64-bp highly conserved consensus sequence cannot very good support to the function as origin and enhancer, and require the assistance of other cis-elements in hrs.

  14. Antifeedant, larvicidal and growth inhibitory bioactivities of novel polyketide metabolite isolated from Streptomyces sp. AP-123 against Helicoverpa armigera and Spodoptera litura

    PubMed Central

    2013-01-01

    Background Considerable attention has been paid to actinomycetes, especially the secondary metabolites obtained from Streptomyces species, as the best alternatives to chemicals as biological control agents for polyphagous pests such as Helicoverpa armigera and Spodoptera litura. On the basis of their novel biocontrol attributes, novel polyketide metabolite isolated from marine Streptomyces sp. AP-123 exhibited significant antifeedant, larvicidal and growth inhibitory activities against polyphagous pests. Results Leaf disc no-choice method was used for the insect bioassay. The polyketide metabolite presented significant antifeedant activities against H. armigera (78.51%) and S. litura (70.75%) at 1000 ppm concentration. The metabolite also exhibited high larvicidal activities against H. armigera (63.11%) and S. litura (58.22%) and the LC50 values were 645.25 ppm for H. armigera and 806.54 ppm for S. litura. The metabolite also prolonged the larval–pupal duration of the insects at all the tested concentrations. Conclusions The activities of the polyketide metabolite were concentration dependent for both the insects therefore it could be used as an agent to prepare new pesticidal formulations. PMID:23668716

  15. Occurrence of a common binding site in Mamestra brassicae, Phthorimaea operculella, and Spodoptera exigua for the insecticidal crystal proteins CryIA from Bacillus thuringiensis.

    PubMed

    Escriche, B; Ferré, J; Silva, F J

    1997-07-01

    Specific binding to midgut membrane proteins is required for the toxicity of insecticidal crystal proteins (ICP) from Bacillus thuringiensis. A direct relationship between toxicity and binding has been proposed. It has been hypothesized that sharing of a single receptor by more than one ICP could lead to the occurrence of multiple resistance in the event of an alteration in the common receptor. Binding of CryIA(a), CryIA(b) and CryIA(c), three structurally related ICPs, has been studied in Phthorimaea operculella, Mamestra brassicae and, Spodoptera exigua using brush border membrane vesicles (BBMV) from the midgut tissue. Using iodinated CryIA(b), the three insects showed similar results: one binding site for CryIA(b), which is shared with CryIA(a) and CryIA(c). The binding site concentrations obtained for CryIA(b) in P. operculella, M. brassicae and S. exigua were 5.1, 16.3 and 2.2 pmol/mg vesicle protein, respectively. In the same way, dissociation constants were 3.8, 5.3 and 0.7 nM. Data show that binding for an ICP does not directly imply toxicity. The occurrence of a common receptor for the CryIA subgroup of ICPs in P. operculella, M. brassicae and S. exigua might theoretically discourage the use of combinations of these ICPs in integrated pest management programmes. PMID:9404010

  16. Cadherin is involved in the action of Bacillus thuringiensis toxins Cry1Ac and Cry2Aa in the beet armyworm, Spodoptera exigua.

    PubMed

    Qiu, Lin; Hou, Leilei; Zhang, Boyao; Liu, Lang; Li, Bo; Deng, Pan; Ma, Weihua; Wang, Xiaoping; Fabrick, Jeffrey A; Chen, Lizhen; Lei, Chaoliang

    2015-05-01

    Bacillus thuringiensis (Bt) insecticidal crystal (Cry) proteins are effective against some insect pests in sprays and transgenic crops, although the evolution of resistance could threaten the long-term efficacy of such Bt use. One strategy to delay resistance to Bt crops is to "pyramid" two or more Bt proteins that bind to distinct receptor proteins within the insect midgut. The most common Bt pyramid in cotton (Gossypium hirsutum L.) employs Cry1Ac with Cry2Ab to target several key lepidopteran pests, including the beet armyworm, Spodoptera exigua (Hübner), which is a serious migratory pest of many vegetable crops and is increasingly important in cotton in China. While cadherin and aminopeptidase-N are key receptors of Cry1 toxins in many lepidopterans including S. exigua, the receptor for Cry2A toxins remains poorly characterized. Here, we show that a heterologous expressed peptide corresponding to cadherin repeat 7 to the membrane proximal extracellular domain (CR7-MPED) in the S. exigua cadherin 1b (SeCad1b) binds Cry1Ac and Cry2Aa. Moreover, SeCad1b transcription was suppressed in S. exigua larvae by oral RNA interference and susceptibility to Cry1Ac and Cry2Aa was significantly reduced. These results indicate that SeCad1b plays important functional roles of both Cry1Ac and Cry2Aa, having major implications for resistance management for S. exigua in Bt crops. PMID:25754522

  17. The impact of the Bacillus subtilis SPB1 biosurfactant on the midgut histology of Spodoptera littoralis (Lepidoptera: Noctuidae) and determination of its putative receptor.

    PubMed

    Ghribi, Dhouha; Abdelkefi-Mesrati, Lobna; Boukedi, Hanen; Elleuch, Mouna; Ellouze-Chaabouni, Semia; Tounsi, Slim

    2012-02-01

    SPB1 is a Bacillus subtilis strain producing a lipopeptide biosurfactant. The insecticidal activity of this biosurfactant was evaluated against the Egyptian cotton leaf worm (Spodoptera littoralis). It displayed toxicity with an LC(50) of 251 ng/cm(2). The histopathological changes occurred in the larval midgut of S. littoralis treated with B. subtilis SPB1 biosurfactant were vesicle formation in the apical region, cellular vacuolization and destruction of epithelial cells and their boundaries. Ligand-blotting experiments with S. littoralis brush border membrane vesicles showed binding of SPB1 biosurfactant to a protein of 45 kDa corresponding to its putative receptor. The latter differs in molecular size from those recognized by Bacillus thuringiensis Vip3A and Cry1C toxins, commonly known by their activity against S. littoralis. This result wires the application of B. subtilis biosurfactant for effective control of S. littoralis larvae, particularly in the cases where S. littoralis will develop resistance against B. thuringiensis toxins. PMID:22079884

  18. Colonization of the Intestinal Tract of the Polyphagous Pest Spodoptera littoralis with the GFP-Tagged Indigenous Gut Bacterium Enterococcus mundtii

    PubMed Central

    Teh, Beng-Soon; Apel, Johanna; Shao, Yongqi; Boland, Wilhelm

    2016-01-01

    The alkaline gut of Lepidopterans plays a crucial role in shaping communities of bacteria. Enterococcus mundtii has emerged as one of the predominant gut microorganisms in the gastrointestinal tract of the major agricultural pest, Spodoptera littoralis. Therefore, it was selected as a model bacterium to study its adaptation to harsh alkaline gut conditions in its host insect throughout different stages of development (larvae, pupae, adults, and eggs). To date, the mechanism of bacterial survival in insects' intestinal tract has been unknown. Therefore, we have engineered a GFP-tagged species of bacteria, E. mundtii, to track how it colonizes the intestine of S. littoralis. Three promoters of different strengths were used to control the expression of GFP in E. mundtii. The promoter ermB was the most effective, exhibiting the highest GFP fluorescence intensity, and hence was chosen as our main construct. Our data show that the engineered fluorescent bacteria survived and proliferated in the intestinal tract of the insect at all life stages for up to the second generation following ingestion. PMID:27379058

  19. Development and reproduction of Spodoptera eridania (Lepidoptera: Noctuidae) and its egg parasitoid Telenomus remus (Hymenoptera: Platygastridae) on the genetically modified soybean (Bt) MON 87701×MON 89788.

    PubMed

    Bortolotto, O C; Silva, G V; de Freitas Bueno, A; Pomari, A F; Martinelli, S; Head, G P; Carvalho, R A; Barbosa, G C

    2014-12-01

    Genetically modified crops with insect resistance genes from Bacillus thuringiensis Berliner (Bt-plants) are increasingly being cultivated worldwide. Therefore, it is critical to improve our knowledge of their direct or indirect impact not only on target pests but also on non-target arthropods. Hence, this study evaluates comparative leaf consumption and performance of Spodoptera eridania (Cramer), a species that is tolerant of the Cry1Ac protein, fed with Bt soybean, MON 87701×MON 89788 or its near [corrected] non-Bt isoline. Using this species as a model, we assessed [corrected] the comparative performance of the egg parasitoid Telenomus remus Nixon on eggs of S. eridania produced from individuals that fed on these two soybean genotypes [corrected] as larvae. Results showed that Bt soybean did not affect pest foliage consumption, but did reduce larvel duration by two days despite larvae in both treatments having six instars. Nevertheless, survival of S. eridania larvae, pupal weight, sex ratio, fecundity and longevity of female moths, and egg viability did not differ between Bt and non-Bt soybeans. Adult longevity of S. eridania males was increased when caterpillars were fed with Bt soybean versus the near isoline. No adverse effects of this technology were observed for the egg parasitoid T. remus. [corrected]. PMID:25248849

  20. Transcriptional profiling analysis of Spodoptera litura larvae challenged with Vip3Aa toxin and possible involvement of trypsin in the toxin activation

    PubMed Central

    Song, Feifei; Chen, Chen; Wu, Songqing; Shao, Ensi; Li, Mengnan; Guan, Xiong; Huang, Zhipeng

    2016-01-01

    Vip proteins, a new group of insecticidal toxins produced by Bacillus thuringiensis, are effective against specific pests including Spodoptera litura. Here, we report construction of a transcriptome database of S. litura by de novo assembly along with detection of the transcriptional response of S. litura larvae to Vip3Aa toxin. In total, 56,498 unigenes with an N50 value of 1,853 bp were obtained. Results of transcriptome abundance showed that Vip3Aa toxin provoked a wide transcriptional response of the S. litura midgut. The differentially expressed genes were enriched for immunity-related, metabolic-related and Bt-related genes. Twenty-nine immunity-related genes, 102 metabolic-related genes and 62 Bt-related genes with differential expression were found. On the basis of transcriptional profiling analysis, we focus on the functional validation of trypsin which potentially participated in the activation of Vip3Aa protoxin. Zymogram analysis indicated that the presence of many proteases, including trypsin, in S. litura larvae midgut. Results of enzymolysis in vitro of Vip3Aa by trypsin, and bioassay and histopathology of the trypsin-digested Vip3Aa toxin showed that trypsin was possibly involved in the Vip3Aa activation. This study provides a transcriptome foundation for the identification and functional validation of the differentially expressed genes in an agricultural important pest, S. litura. PMID:27025647

  1. Performance of Spodoptera litura Fabricius on different host plants: influence of nitrogen and total phenolics of plants and mid-gut esterase activity of the insect.

    PubMed

    Ghumare, S S; Mukherjee, S N

    2003-08-01

    Five host plants [castor, Ricinus communis (Carolus Linnaeus); cotton, Gossypium hirsutm (Carolus Linnaeus); tomato, Lycopersicum esculentum (Philip Miller); mint, Mentha arvensis (Carolus Linnaeus) and cabbage, Brassica oleracea (Carolus Linnaeus)] belonging to different families were used to study the performance of the Asian armyworm, Spodoptera litura larvae. Highest consumption of food and dry weight gain was observed in larvae fed on castor. Mint did not support optimum larval growth because of low digestibility and low efficiency of conversion of digested food to body matter. Dry weight gain ranged from 26.64 mg on mint to 86.80 mg in castor. These differences tend to be related to nitrogen and total phenolics content of the leaf tissues; however, the most clear-cut correlation is an inverse one between the host plant preference and the ratio of total phenolics to nitrogen in the leaf tissues. Mid-gut esterase activity in larvae showed an increasing trend with the increase in total phenolics: nitrogen ratio in the test plants and the order of mid-gut esterase activity in larvae was mint > cabbage > cotton > tomato > castor. PMID:15248492

  2. A Transformed Bacterium Expressing Double-Stranded RNA Specific to Integrin β1 Enhances Bt Toxin Efficacy against a Polyphagous Insect Pest, Spodoptera exigua

    PubMed Central

    Kim, Eunseong; Park, Youngjin; Kim, Yonggyun

    2015-01-01

    Background Oral toxicity of double-stranded RNA (dsRNA) specific to integrin β1 subunit (SeINT) was known in a polyphagous insect pest, Spodoptera exigua. For an application of the dsRNA to control the insect pest, this study prepared a transformed Escherichia coli expressing dsRNA specific to SeINT. Principal Findings The dsRNA expression was driven by T7 RNA polymerase overexpressed by an inducer in the transformed E. coli. The produced dsRNA amount was proportional to the number of the cultured bacteria. The transformed bacteria gave a significant oral toxicity to S. exigua larvae with a significant reduction of the SeINT expression. The resulting insect mortality increased with the fed number of the bacteria. Pretreatment with an ultra-sonication to disrupt bacterial cell wall/membrane significantly increased the insecticidal activity of the transformed bacteria. The larvae treated with the transformed bacteria suffered tissue damage in the midgut epithelium, which exhibited a marked loss of cell-cell contacts and underwent a remarkable cell death. Moreover, these treated larvae became significantly susceptible to a Cry toxin derived from Bacillus thuringiensis (Bt). Conclusions This study provides a novel and highly efficient application technique to use dsRNA specific to an integrin gene by mixing with a biopesticide, Bt. PMID:26171783

  3. Parasitic Manipulation of Host Behaviour: Baculovirus SeMNPV EGT Facilitates Tree-Top Disease in Spodoptera exigua Larvae by Extending the Time to Death

    PubMed Central

    Han, Yue; van Houte, Stineke; Drees, Gerben F.; van Oers, Monique M.; Ros, Vera I. D.

    2015-01-01

    Many parasites enhance their dispersal and transmission by manipulating host behaviour. One intriguing example concerns baculoviruses that induce hyperactivity and tree-top disease (i.e., climbing to elevated positions prior to death) in their caterpillar hosts. Little is known about the underlying mechanisms of such parasite-induced behavioural changes. Here, we studied the role of the ecdysteroid UDP-glucosyltransferase (egt) gene of Spodoptera exigua multiple nucleopolyhedrovirus (SeMNPV) in tree-top disease in S. exigua larvae. Larvae infected with a mutant virus lacking the egt gene exhibited a shorter time to death and died before the induction of tree-top disease. Moreover, deletion of either the open reading frame or the ATG start codon of the egt gene prevented tree-top disease, indicating that the EGT protein is involved in this process. We hypothesize that SeMNPV EGT facilitates tree-top disease in S. exigua larvae by prolonging the larval time to death. Additionally, we discuss the role of egt in baculovirus-induced tree-top disease. PMID:26463412

  4. Impact of UV radiation on activity of linear furanocoumarins and Bacillus thuringiensis var. kurstaki against Spodoptera exigua: Implications for tritrophic interactions

    SciTech Connect

    Trumble, J.T.; Moar, W.J.; Brewer, M.J.; Carson, W.G. )

    1991-05-01

    Acidic fogs with a pH of 2.0 and duration of 2 hr did not reduce the efficacy of Bacillus thuringiensis var. kurstaki. Therefore, the impact of UV radiation was investigated on the interactions between (1) levels of the antibacterial linear furanocoumarins psoralen, bergapten, and xanthotoxin in Apium graveolens (L.) occurring following a 2.0 pH acidic fog episode, (2) the noctuid Spodoptera exigua, and (3) a sublethal dosage of the microbial pathogen B. thuringiensis var. kurstaki. Mean time to pupation in the absence of UV radiation was significantly extended by the addition of either psoralens or B. thuringiensis. Larvae developing on diets containing B. thuringiensis plus psoralens required nearly 40% longer to pupate than controls, but their effects were additive as the interaction was not significant. Mean time to mortality, a weighted average time of death, was not significantly affected by any of the treatments. In a 2 {times} 2 {times} 2 factorial analysis, all main effects reduced survival significantly, as did the three-way interaction. Thus, antagonistic interactions with psoralens that would reduce the effectiveness of B. thuringiensis in the field were not observed. When pairs of main effects were nested within the two levels of the third factor, several two-way interactions were found. Interestingly, the activity of B. thuringiensis and the psoralens, individually or in combination, was enhanced by exposure to UV radiation. Implications of this research are discussed for both natural and agricultural ecosystems.

  5. Parasitic Manipulation of Host Behaviour: Baculovirus SeMNPV EGT Facilitates Tree-Top Disease in Spodoptera exigua Larvae by Extending the Time to Death.

    PubMed

    Han, Yue; van Houte, Stineke; Drees, Gerben F; van Oers, Monique M; Ros, Vera I D

    2015-01-01

    Many parasites enhance their dispersal and transmission by manipulating host behaviour. One intriguing example concerns baculoviruses that induce hyperactivity and tree-top disease (i.e., climbing to elevated positions prior to death) in their caterpillar hosts. Little is known about the underlying mechanisms of such parasite-induced behavioural changes. Here, we studied the role of the ecdysteroid UDP-glucosyltransferase (egt) gene of Spodoptera exigua multiple nucleopolyhedrovirus (SeMNPV) in tree-top disease in S. exigua larvae. Larvae infected with a mutant virus lacking the egt gene exhibited a shorter time to death and died before the induction of tree-top disease. Moreover, deletion of either the open reading frame or the ATG start codon of the egt gene prevented tree-top disease, indicating that the EGT protein is involved in this process. We hypothesize that SeMNPV EGT facilitates tree-top disease in S. exigua larvae by prolonging the larval time to death. Additionally, we discuss the role of egt in baculovirus-induced tree-top disease. PMID:26463412

  6. Identification and Characterization of CYP9A40 from the Tobacco Cutworm Moth (Spodoptera litura), a Cytochrome P450 Gene Induced by Plant Allelochemicals and Insecticides

    PubMed Central

    Wang, Rui-Long; Staehelin, Christian; Xia, Qing-Qing; Su, Yi-Juan; Zeng, Ren-Sen

    2015-01-01

    Cytochrome P450 monooxygenases (P450s) of insects play crucial roles in the metabolism of endogenous and dietary compounds. Tobacco cutworm moth (Spodoptera litura), an important agricultural pest, causes severe yield losses in many crops. In this study, we identified CYP9A40, a novel P450 gene of S. litura, and investigated its expression profile and potential role in detoxification of plant allelochemicals and insecticides. The cDNA contains an open reading frame encoding 529 amino acid residues. CYP9A40 transcripts were found to be accumulated during various development stages of S. litura and were highest in fifth and sixth instar larvae. CYP9A40 was mainly expressed in the midgut and fat body. Larval consumption of xenobiotics, namely plant allelochemicals (quercetin and cinnamic acid) and insecticides (deltamethrin and methoxyfenozide) induced accumulation of CYP9A40 transcripts in the midgut and fat body. Injection of dsCYP9A40 (silencing of CYP9A40 by RNA interference) significantly increased the susceptibility of S. litura larvae to the tested plant allelochemicals and insecticides. These results indicate that CYP9A40 expression in S. litura is related to consumption of xenobiotics and suggest that CYP9A40 is involved in detoxification of these compounds. PMID:26393579

  7. Imperfection works: Survival, transmission and persistence in the system of Heliothis virescens ascovirus 3h (HvAV-3h), Microplitis similis and Spodoptera exigua.

    PubMed

    Li, Shun-Ji; Hopkins, Richard J; Zhao, Yi-Pei; Zhang, Yun-Xuan; Hu, Jue; Chen, Xu-Yang; Xu, Zhi; Huang, Guo-Hua

    2016-01-01

    Ascoviruses are insect-specific large DNA viruses that mainly infect noctuid larvae, and are transmitted by parasitoids in the fields. Heliothis virescens ascovirus 3h (HvAV-3h) has been recently isolated from Spodoptera exigua, without parasitoid vector identified previously. Here we report that Microplitis similis, a solitary endoparasitoid wasp, could transmit HvAV-3h between S. exigua larvae in the laboratory. When the female parasitoid wasp acquired the virus and served as a vector, the period of virion viability on the ovipositor was 4.1 ± 1.4 days. Infected host larvae were still acceptable for egg laying by parasitoids, and the parasitoids thereafter transmitted virus to healthy hosts. Virus acquisition occurred only from donor hosts between 3 and 9 days post infection. The peak of virus acquisition (80.9 ± 6.3%) was found when M. similis wasps oviposited in larvae that had been inoculated with the virus 7 days previously. When virus infection of the host took place during the life cycle of the parasitoid wasp, it caused 1- to 4-day-old immature parasitoids death in the host, whilst a small proportion of 5- to 6-day-old and the majority of 7-day-old parasitoids larvae survived from the virus-infected hosts. Viral contamination did not reduce the life span or fecundity of female M. similis. PMID:26878829

  8. Transcriptional profiling analysis of Spodoptera litura larvae challenged with Vip3Aa toxin and possible involvement of trypsin in the toxin activation.

    PubMed

    Song, Feifei; Chen, Chen; Wu, Songqing; Shao, Ensi; Li, Mengnan; Guan, Xiong; Huang, Zhipeng

    2016-01-01

    Vip proteins, a new group of insecticidal toxins produced by Bacillus thuringiensis, are effective against specific pests including Spodoptera litura. Here, we report construction of a transcriptome database of S. litura by de novo assembly along with detection of the transcriptional response of S. litura larvae to Vip3Aa toxin. In total, 56,498 unigenes with an N50 value of 1,853 bp were obtained. Results of transcriptome abundance showed that Vip3Aa toxin provoked a wide transcriptional response of the S. litura midgut. The differentially expressed genes were enriched for immunity-related, metabolic-related and Bt-related genes. Twenty-nine immunity-related genes, 102 metabolic-related genes and 62 Bt-related genes with differential expression were found. On the basis of transcriptional profiling analysis, we focus on the functional validation of trypsin which potentially participated in the activation of Vip3Aa protoxin. Zymogram analysis indicated that the presence of many proteases, including trypsin, in S. litura larvae midgut. Results of enzymolysis in vitro of Vip3Aa by trypsin, and bioassay and histopathology of the trypsin-digested Vip3Aa toxin showed that trypsin was possibly involved in the Vip3Aa activation. This study provides a transcriptome foundation for the identification and functional validation of the differentially expressed genes in an agricultural important pest, S. litura. PMID:27025647

  9. Gasmin (BV2-5), a polydnaviral-acquired gene in Spodoptera exigua. Trade-off in the defense against bacterial and viral infections.

    PubMed

    Gasmi, Laila; Jakubowska, Agata K; Herrero, Salvador

    2016-03-01

    Thousands of Hymenopteran endoparasitoids have developed a unique symbiotic relationship with viruses named polydnavirus (PDVs). These viruses immunocompromise the lepidopteran host allowing the survival of the wasp eggs. In a previous work, we have shown the horizontal transfer of some polydnaviral genes into the genome of the Lepidoptera, Spodoptera exigua. One of these genes, BV2-5 (named gasmin) interferes with actin polymerization, negatively affecting the multiplication of baculovirus in cell culture. In this work, we have focused in the study of the effect of Gasmin expression on different aspects of the baculovirus production. In addition, and since actin polymerization is crucial for phagocytosis, we have studied the effect of Gasmin expression on the larval interaction with bacterial pathogens. Over-expression of Gasmin on hemocytes significantly reduces their capacity to phagocytize the pathogenic bacteria Bacillus thuringiensis. According to these results, gasmin domestication negatively affects baculovirus replication, but increases larvae susceptibility to bacterial infections as pay off. Although the effect of Gasmin on the insect interaction with other pathogens or parasitoids remain unknown, the opposite effects described here could shape the biological history of this species based on the abundance of certain type of pathogens as suggested by the presence of truncated forms of this protein in several regions of the world. PMID:26658027

  10. RNA interference of a heat shock protein, Hsp70, loses its protection role in indirect chilling injury to the beet armyworm, Spodoptera exigua.

    PubMed

    Choi, Bong-Gee; Hepat, Rahul; Kim, Yonggyun

    2014-02-01

    The beet armyworm, Spodoptera exigua, is freeze-susceptible, in which glycerol plays a crucial role in depressing supercooling point (SCP) to avoid the freezing injury. This study focused on a non-freezing injury classified into indirect chilling injury of S. exigua after a prolonged exposure to low temperatures much above SCPs. Exposure to 0 and 5°C for longer than 2weeks was lethal to all the immature stages. Among immature stages, eggs were the most susceptible to the low temperature treatments and pupae were the next susceptible. Among larvae, the third instar (L3) appeared to be more tolerant than the fifth instar (L5). The temperature treatment at 15°C allowed both L3 and L5 to exhibit a feeding behavior and induced little non-freezing injury, suggesting a minimal temperature threshold for optimal overwintering conditions of S. exigua. Three heat shock protein genes (Hsp70, Hsp74, Hsp83) were expressed in the larvae at the low temperature treatments. Only Hsp70 was inducible to the low temperatures in both L3 and L5 stages. RNA interference of Hsp70 expression led to significantly lose the survival rates of the treated larvae in the conditions inducing the non-freezing injury. These results suggest that Hsp70 plays a role in protecting S. exigua from the indirect chilling injury. PMID:24309290

  11. Effect of the Extracts of the Spiderflower, Cleome arabica, on Feeding and Survival of Larvae of the Cotton Leafworm, Spodoptera littoralis

    PubMed Central

    Ladhari, Afef; Laarif, Asma; Omezzine, Faten; Haouala, Rabiaa

    2013-01-01

    Aqueous and organic (hexane, chloroform, and methanol) extracts of siliquae, stems and leaves, and seeds of Cleome arabica L. (Brassicales: Capparidaceae) were evaluated in the laboratory for their antifeeding and insecticidal effect on larvae of the cotton leafworm, Spodoptera littoralis (Boisduval) (Lepidoptera, Noctuidae), using a leaf dipping bioassay with castor bean, Ricinus communis L. (Malpighiales: Euphorbiaceae), leaf discs. The polar extracts caused significant mortality. At the highest dose, C. arabica extracts exhibited significant antifeeding and phagostimulating activities against S. littoralis larvae. Under no-choice conditions, the methanol extract of siliquae was the most active, and the antifeedant index calculated over 24 hr for 3rd instar larvae varied significantly from 16 to 37%. Using nutritional indices, it was established that there was a significant decrease in growth rate concomitant with a reduction in consumption. These results suggest the presence of anti-feeding and/or toxic substances in the extracts that may be useful in developing bio-insecticides based on C. arabica extracts for use in integrated pest management of leafworm and other agricultural pests. PMID:23906290

  12. [The gene expression of the protein SLAWD, mediating the toxic effect of destruxin A on Spodoptera litura larvae, in procaryotic cells: purification and characterization].

    PubMed

    Meng, X; Li, L M; Gao, G; Jin, F L; Ren, S X

    2014-01-01

    Spodoptera litura is one of the most destructive phytophagous pest infesting cotton, vegetable, oilseed and ber crops around the world. Dextruxin A (DA), is a one of a kind microbial insecticide, which has potent toxins with bioactivity against S. litura larvae. An abnormal wing disc (AWD) protein was identified as a DA toxic effect protein in S. litura SL-1 cells. To better understand the role of the AWD gene of S. litura (SLAWD) it was purified and characterized. The entire coding region of the SLAWD gene was cloned into a pET-32a(+) expression vector and transformed into competent Escherichia coli BL21 (DE3) cells. SDS-PAGE and western blotting analysis and western blotting showed that the best induction conditions were 1 mmol mL(-1) isopropyl-β-D-thiogalactopyranoside (IPTG) for 6 h at 37°C; the molecular weight of the fusion protein was 35.0 kDa. The production of polyclonal antibodies and an enzyme-linked immunosorbent assay (ELISA) showed that the titer of antiserum was 1:25,600; western blotting analysis showed that the recombinant SLAWD was recognized by the anti-SLAWD polyclonal antibody. AWD is a key protein involved in wing development in insects. These tools will assist in the further characterization of SLAWD and studies on the mechanism of action of destruxin A. PMID:25845231

  13. RNAi-mediated knockdown of catalase causes cell cycle arrest in SL-1 cells and results in low survival rate of Spodoptera litura (Fabricius).

    PubMed

    Zhao, Haiming; Yi, Xin; Hu, Zhen; Hu, Meiying; Chen, Shaohua; Muhammad, Rizwan-ul-Haq; Dong, Xiaolin; Gong, Liang

    2013-01-01

    Deregulated reactive oxygen species (ROS) production can lead to the disruption of structural and functional integrity of cells as a consequence of reactive interaction between ROS and various biological components. Catalase (CAT) is a common enzyme existing in nearly all organisms exposed to oxygen, which decomposes harmful hydrogen peroxide, into water and oxygen. In this study, the full length sequence that encodes CAT-like protein from Spodoptera litura named siltCAT (GenBank accession number: JQ_663444) was cloned and characterized. Amino acid sequence alignment showed siltCAT shared relatively high conservation with other insect, especially the conserved residues which defined heme and NADPH orientation. Expression pattern analysis showed that siltCAT mRNA was mainly expressed in the fat body, midgut, cuticle and malpighian tube, and as well as over last instar larvae, pupa and adult stages. RNA interference was used to silence CAT gene in SL-1 cells and the fourth-instar stage of S. litura larvae respectively. Our results provided evidence that CAT knockdown induced ROS generation, cell cycle arrest and apoptosis in SL-1 cells. It also confirmed the decrease in survival rate because of increased ROS production in experimental groups injected with double-stranded RNA of CAT (dsCAT). This study implied that ROS scavenging by CAT is important for S. litura survival. PMID:23555693

  14. Identification and Characterization of CYP9A40 from the Tobacco Cutworm Moth (Spodoptera litura), a Cytochrome P450 Gene Induced by Plant Allelochemicals and Insecticides.

    PubMed

    Wang, Rui-Long; Staehelin, Christian; Xia, Qing-Qing; Su, Yi-Juan; Zeng, Ren-Sen

    2015-01-01

    Cytochrome P450 monooxygenases (P450s) of insects play crucial roles in the metabolism of endogenous and dietary compounds. Tobacco cutworm moth (Spodoptera litura), an important agricultural pest, causes severe yield losses in many crops. In this study, we identified CYP9A40, a novel P450 gene of S. litura, and investigated its expression profile and potential role in detoxification of plant allelochemicals and insecticides. The cDNA contains an open reading frame encoding 529 amino acid residues. CYP9A40 transcripts were found to be accumulated during various development stages of S. litura and were highest in fifth and sixth instar larvae. CYP9A40 was mainly expressed in the midgut and fat body. Larval consumption of xenobiotics, namely plant allelochemicals (quercetin and cinnamic acid) and insecticides (deltamethrin and methoxyfenozide) induced accumulation of CYP9A40 transcripts in the midgut and fat body. Injection of dsCYP9A40 (silencing of CYP9A40 by RNA interference) significantly increased the susceptibility of S. litura larvae to the tested plant allelochemicals and insecticides. These results indicate that CYP9A40 expression in S. litura is related to consumption of xenobiotics and suggest that CYP9A40 is involved in detoxification of these compounds. PMID:26393579

  15. A mode of action of glucosinolate-derived isothiocyanates: Detoxification depletes glutathione and cysteine levels with ramifications on protein metabolism in Spodoptera littoralis.

    PubMed

    Jeschke, Verena; Gershenzon, Jonathan; Vassão, Daniel Giddings

    2016-04-01

    Glucosinolates are activated plant defenses common in the order Brassicales that release isothiocyanates (ITCs) and other hydrolysis products upon tissue damage. The reactive ITCs are toxic to insects resulting in reduced growth, delayed development and occasionally mortality. Generalist lepidopteran larvae often detoxify ingested ITCs via conjugation to glutathione (GSH) and survive on low glucosinolate diets, but it is not known how this process influences other aspects of metabolism. We investigated the impact of the aliphatic 4-methylsulfinylbutyl-ITC (4msob-ITC, sulforaphane) on the metabolism of Spodoptera littoralis larvae, which suffer a significant growth decline on 4msob-ITC-containing diets while excreting ITC-glutathione conjugates and their derivatives in the frass. The most striking effects were a decrease of GSH in midgut tissue and hemolymph due to losses by conjugation to ITC during detoxification, and a decline of the GSH biosynthetic precursor cysteine. Protein content was likewise reduced by ITC treatment suggesting that protein is actively catabolized in an attempt to supply cysteine for GSH biosynthesis. The negative growth and protein effects were relieved by dietary supplementation with cystine. Other consequences of protein breakdown included deamination of amino acids with increased excretion of uric acid and elevated lipid content. Thus metabolic detoxification of ITCs provokes a cascade of negative effects on insects that result in reduced fitness. PMID:26855197

  16. The effect of leaf biopesticide Mirabilis jalapa and fungi Metarhizium anisopliae to immune response and mortality of Spodoptera exigua instar IV

    NASA Astrophysics Data System (ADS)

    Suryani, A. Irma; Anggraeni, Tjandra

    2014-03-01

    Spodoptera exigua is one of insect causing damage in agriculture sector. This insect can be controlled by a natural biopesticide by combining two agents of biological control, biopesticides Mirabilis jalapa and entomopathogenic fungi Metarhizium anisopliae, considered to be virulent toward a wide range of insects. The objective of research was to determine the effect of biopesticides M. jalapa and fungi M. anisopliae against immune system and mortality of S. exigua. This research used a complete randomized block design with five concentrations Mirabilis jalapa and optimum dose of M. anisopliae. A high dose of M. jalapa (0.8% w/v) is the most effective one to decrease total haemocytes especially granulocyt and plasmatocyt (cellular immune) and decrease the concentration of lectin (humoral immune) from S. exigua (p < 0.05). The combination of M. jalapa (0, 8% w/v) and lethal dose of M. anisopliae 2.59 × 107 spore/ml were significant to increase mortality of S. exigua within 48 hours (p < 0.05).

  17. Interaction of Gene-Cloned and Insect Cell-Expressed Aminopeptidase N of Spodoptera litura with Insecticidal Crystal Protein Cry1C

    PubMed Central

    Agrawal, Neema; Malhotra, Pawan; Bhatnagar, Raj K.

    2002-01-01

    Insecticidal toxins produced by Bacillus thuringiensis interact with specific receptors located in the midguts of susceptible larvae, and the interaction is followed by a series of biochemical events that lead to the death of the insect. In order to elucidate the mechanism of action of B. thuringiensis toxins, receptor protein-encoding genes from many insect species have been cloned and characterized. In this paper we report the cloning, expression, and characterization of Cry toxin-interacting aminopeptidase N (APN) isolated from the midgut of a polyphagous pest, Spodoptera litura. The S. litura APN cDNA was expressed in the Sf21 insect cell line by using a baculovirus expression system. Immunofluorescence staining of the cells revealed that the expressed APN was located at the surface of Sf21 cells. Treatment of Sf21 cells expressing S. litura APN with phosphatidylinositol-specific phospholipase C demonstrated that the APN was anchored in the membrane by a glycosylphosphatidylinositol moiety. Interaction of the expressed receptor with different Cry toxins was examined by immunofluorescence toxin binding studies and ligand blot and immunoprecipitation analyses. By these experiments we showed that the bioactive toxin, Cry1C, binds to the recombinant APN, while the nonbioactive toxin, Cry1Ac, showed no interaction. PMID:12200317

  18. Evolution of NADPH-cytochrome P450 oxidoreductases (POR) in Apiales - POR 1 is missing.

    PubMed

    Andersen, Trine Bundgaard; Hansen, Niels Bjørn; Laursen, Tomas; Weitzel, Corinna; Simonsen, Henrik Toft

    2016-05-01

    The NADPH-dependent cytochrome P450 oxidoreductase (POR) is the obligate electron donor to eukaryotic microsomal cytochromes P450 enzymes. The number of PORs within plant species is limited to one to four isoforms, with the most common being two PORs per plant. These enzymes provide electrons to a huge number of different cytochromes P450s (from 50 to several hundred within one plant). Within the eudicotyledons, PORs can be divided into two major clades, POR 1 and POR 2. Based on our own sequencing analysis and publicly available data, we have identified 45 PORs from the angiosperm order Apiales. These were subjected to a phylogenetic analysis along with 237 other publicly available (NCBI and oneKP) POR sequences found within the clade Asterids. Here, we show that the order Apiales only harbor members of the POR 2 clade, which are further divided into two distinct subclades. This is in contrast to most other eudicotyledon orders that have both POR 1 and POR 2. This suggests that through gene duplications and one gene deletion, Apiales only contain members of the POR 2 clade. Three POR 2 isoforms from Thapsia garganica L., Apiaceae, were all full-length in an Illumina root transcriptome dataset (available from the SRA at NCBI). All three genes were shown to be functional upon reconstitution into nanodiscs, confirming that none of the isoforms are pseudogenes. PMID:26854662

  19. Efficacy of some plant oils alone and/or combined with different insecticides on the cotton leaf-worm Spodoptera littoralis (Boisd.) (Lepidoptera: Noctuidae) in Egypt.

    PubMed

    Mesbah, H A; Mourad, A K; Rokaia, A Z M

    2006-01-01

    The present work was conducted to evaluate the efficiency of two essential oils ((Flax or "Linseed" and Sesame ), five volatile plant oils ( Camphor, Red basil, Rose, Menthol and Clove ), four pesticides (Methoxyfenozide; Permethrin; Profenofos and Spinosad) and their mixtures on the cotton leaf-worm Spodoptera littoralis (Boisd.). This study was also devoted to minimize the usage of conventional insecticides, reduce the environmental pollution, and protect human-beings and domestic animals from hazards due to pesticides applications. In the meantime, the delayed effect of these tested plant oils on the developing immatures and moths of the cotton leaf-worm, Spodoptera littoralis was determined. Most of the evaluated plant oils were found to have an insecticidal effect on the 4th instar larvae of the cotton leaf-worm, S. littoralis. Both the essential Sesame oil and Clove volatile oil showed rather weak toxic effect corresponding to the same concentrations and periods of the bioassay tests. Comparing the toxicity of the tested plant oils, it was affirmed that both Rose and Red basil volatile oils were the highest efficient natural phytocompounds against the treated larvae and alternatively ranked either the 1st and/or the 2nd rank, throughout the different periods of the bioassay tests, followed by the other three tested oils which were more or less efficient phytocompounds. According to the toxicity index, all the tested oils were less toxic than the superior Red basil volatile oil after 48 h. post treatment followed by the gradual decrease in toxicity of Rose, Flax and Menthol, respectively. The development of the treated 4th larval instar was blocked due to treatment with the tested plant oils. With no exception, all the efficiently tested essential and/or volatile oils acted principally as Insect Growth Inhibitors (IGIs) rather than antifeedants causing disruption of the insect development, abnormal larvae, pupae and adults that were lead finally to death. The

  20. Comprehensive Analysis of Gene Expression Profiles of the Beet Armyworm Spodoptera exigua Larvae Challenged with Bacillus thuringiensis Vip3Aa Toxin

    PubMed Central

    Bel, Yolanda; Jakubowska, Agata K.; Costa, Juliana; Herrero, Salvador; Escriche, Baltasar

    2013-01-01

    Host-pathogen interactions result in complex relationship, many aspects of which are not completely understood. Vip proteins, which are Bacillus thuringensis (Bt) insecticidal toxins produced during the vegetative stage, are selectively effective against specific insect pests. This new group of Bt proteins represents an interesting alternative to the classical Bt Cry toxins because current data suggests that they do not share the same mode of action. We have designed and developed a genome-wide microarray for the beet armyworm Spodoptera exigua, a serious lepidopteran pest of many agricultural crops, and used it to better understand how lepidopteran larvae respond to the treatment with the insecticidal protein Vip3Aa. With this approach, the goal of our study was to evaluate the changes in gene expression levels caused by treatment with sublethal doses of Vip3Aa (causing 99% growth inhibition) at 8 and 24 h after feeding. Results indicated that the toxin provoked a wide transcriptional response, with 19% of the microarray unigenes responding significantly to treatment. The number of up- and down-regulated unigenes was very similar. The number of genes whose expression was regulated at 8 h was similar to the number of genes whose expression was regulated after 24 h of treatment. The up-regulated sequences were enriched for genes involved in innate immune response and in pathogen response such as antimicrobial peptides (AMPs) and repat genes. The down-regulated sequences were mainly unigenes with homology to genes involved in metabolism. Genes related to the mode of action of Bt Cry proteins were found, in general, to be slightly overexpressed. The present study is the first genome-wide analysis of the response of lepidopteran insects to Vip3Aa intoxication. An insight into the molecular mechanisms and components related to Vip intoxication will allow designing of more effective management strategies for pest control. PMID:24312604

  1. Expression analysis of two P450 monooxygenase genes of the tobacco cutworm moth (Spodoptera litura) at different developmental stages and in response to plant allelochemicals.

    PubMed

    Wang, Rui-Long; Li, Jun; Staehelin, Christian; Xin, Xiao-Wei; Su, Yi-Juan; Zeng, Ren-Sen

    2015-01-01

    Cytochrome P450 monooxygenases (P450s) of insects are known to be involved in the metabolism or detoxification of plant allelochemicals and insecticides. Spodoptera litura (Lepidoptera, Noctuidae) is a polyphagous moth responsible for severe yield losses in many crops. In this study, two full-length P450 genes, CYP6B48 and CYP6B58, were cloned from S. litura. The cDNA sequences encode proteins with 503 and 504 amino acids, respectively. Phylogenetic analysis revealed that CYP6B48 and CYP6B58 belong to the CYP6B subfamily of P450s. Quantitative real-time PCR analyses showed that CYP6B48 and CYP6B58 were expressed only at larval stage, but not at pupal and adult stages. The highest levels of transcripts were found in the midguts and fat bodies of the larvae. No expression was detected in the ovary or hemolymph. Feeding with diets containing cinnamic acid, quercetin, or coumarin did not affect expression of CYP6B48. In contrast, diet supplemented with xanthotoxin dramatically increased the levels of CYP6B48 transcript in the midgut and fat bodies. Larvae fed with flavone had high levels of transcript of CYP6B48 in the midgut, whereas only slightly elevated levels were found in the fat bodies. Effects of the tested allelochemicals on CYP6B58 expression were minor. Hence, our findings show that S. litura responds to specific allelochemicals such as xanthotoxin with the accumulation of CYP6B48 transcripts, suggesting that specific signals in the food control the insect's ability to convert toxic allelochemicals to less harmful forms at the transcriptional level. PMID:25547988

  2. The effect of the combination of two biological control agents, Mirabilis jalapa and Bacillus thuringiensis, to Spodoptera litura's immune response and their mortality

    NASA Astrophysics Data System (ADS)

    Maulina, Dina; Anggraeni, Tjandra

    2014-03-01

    Biological control provides a safer alternative to reduce the population of agricultural pest. Mirabilis jalapa is one of many promising biopesticides which contains chemical substances that have a feeding deterrent property against insects. This biopesticide may not kill insect directly but will weaken their overall physiological condition. In this study, we investigated the immune response of common pestSpodoptera litura after exposure of M. jalapa extract. We also used Bacillus thuringiensis (Bt) delta endotoxin (LC50) on 3 hours after exposure of M. jalapa extract to see the synergism properties of both biopesticide agents. Microscopic observation revealed that at least 5 types of haemocyte were found in S. litura. In control group, plasmatocyte were found at 59.98%, prohaemocyte 20.73%, granullar cell 12.74%, oenocytoid 3.33% and spherule cell 3.20%. These proportion was differ significantly in the treatment group. Exposure to 0.1% and 0.2%(w/v) of M. jalapa extract increased the total number of haemocytes as much as 38.08% and 64.15% respectively. In contrast, exposure to 0.4% and 0.8%(w/v) reduced the number of haemocytes to 37.02% and 51.04% respectively. In term of phagocytic activity, the proportion of phagocytosing cells were 47.62% in control group, and in 0.1% and 0.2% (w/v) M. jalapa treatment group the proportion decreased to 28% and 26.88% respectively. In the concentration of 0.4% and 0.8%, phagocytic activity did not occur. Addition of biological agents Bt (LC50 concentration) to see mortality 3 hours after M. jalapa application did not show significant differences. S. litura mortality rate were found only 50%; this suggests that the combination of M. jalapa and Bt biopesticides in 3-hour intervals within 24 hours showed no increase in mortality.

  3. A promising HD133-like strain of Bacillus thuringiensis with dual efficiency to the two Lepidopteran pests: Spodoptera littoralis (Noctuidae) and Ephestia kuehniella (Pyralidae).

    PubMed

    BenFarhat-Touzri, Dalel; Driss, Fatma; Tounsi, Slim

    2016-08-01

    Isolation and identification of new strains with wide variety of target pests is an ever growing field. In this paper, a screening of 260 strains from Tunisian soil samples was conducted by dot-blot and PCR-sequencing analysis. The screening was done on the basis of the possession of cry1D-type genes and was followed by the evaluation of the insecticidal activity against Spodoptera littoralis. BLB250 strain showed an LC50 value (56.2 μg g(-1)) against S. littoralis lower than those of the two Bacillus thuringiensis reference strains HD1 and HD133. An interesting LC50 (167.6 μg g(-1)) was also recorded against Ephestia kuehniella larvae. The strain was, thus, selected because of its qualification to be highly toxic, at once, for both Lepidopteran insects. In vitro time course of proteolytic processing of BLB250 and HD133 protoxins by the gut juices from the two insect larvae revealed that the differences in toxicity against E. kuehniella are most likely attributed to differences in the efficiency of the activation of the corresponding protoxins into toxins. An activation comparative study using commercial proteases suggested that the intestinal proteases of E. kuehniella contain trypsin-like activities. With its high efficiency and toxicity against, at once, two Lepidopteran insects having different susceptibilities towards kurstaki and aizawai subspecies, BLB250 could be useful when developing more efficient and economical B. thuringiensis-based pesticides. PMID:27130040

  4. Developmental response of Spodoptera litura Fab. to treatments of crude volatile oil from Piper betle L. and evaluation of toxicity to earthworm, Eudrilus eugeniae Kinb.

    PubMed

    Vasantha-Srinivasan, Prabhakaran; Senthil-Nathan, Sengottayan; Thanigaivel, Annamalai; Edwin, Edward-Sam; Ponsankar, Athirstam; Selin-Rani, Selvaraj; Pradeepa, Venkatraman; Sakthi-Bhagavathy, Muthiah; Kalaivani, Kandaswamy; Hunter, Wayne B; Duraipandiyan, Veeramuthu; Al-Dhabi, Naif Abdullah

    2016-07-01

    Evaluations of biological effects of (Pb-CVO) the crude volatile oil of Piper betle leaves on the tobacco cutworm Spodoptera litura were conducted. Pb-CVO was subjected to GC-MS analysis and twenty vital compounds were isolated from the betel leaf oil. Pb-CVO was tested at four different concentrations (0.25, 0.5, 1.0 and 1.5%) against S. litura. The treated insects exhibited dose depended mortality. The mortality rate was significantly higher at the 1.0 and 1.5% Pb-CVO. The LC50 (Lethal concentration) were observed at 0.48% Pb-CVO. Larval and pupal durations increased in all treatment concentrations (0.25, 0.3, 0.4 and 0.5%) whereas, pupal weight decreased compared to control. Adult longevity of S. litura was reduced in all treatments but predominantly in the 0.4 and 0.5% Pb-CVO. Correspondingly, mean fecundity rate was reduced at all concentrations compared to control. Histological studies of larvae mid-gut profiles of S. litura were severely damaged in 1.0 and 1.5% and showed abnormalities in mid-gut cells with 0.25 and 0.5% Pb-CVO treatments. Earthworm toxicity illustrated that 0.1% of chemical insecticides (monocrotophos and cypermethrin) varied widely in their contact toxicities compared to 0.5 and 1.0% Pb-CVO and control in both contact filter paper and artificial soil test. These findings suggest that twenty essential compounds of betel leaf oil were significant inhibitors of the development and caused behavioral changes of S. litura. Treatment with betel leaf oil at these concentrations had no adverse effect on earthworm populations. PMID:27135695

  5. Detoxification of insecticides, allechemicals and heavy metals by glutathione S-transferase SlGSTE1 in the gut of Spodoptera litura.

    PubMed

    Xu, Zhi-Bin; Zou, Xiao-Peng; Zhang, Ni; Feng, Qi-Li; Zheng, Si-Chun

    2015-08-01

    Insect glutathione S-transferases (GSTs) play important roles in detoxifying toxic compounds and eliminating oxidative stress caused by these compounds. In this study, detoxification activity of the epsilon GST SlGSTE1 in Spodoptera litura was analyzed for several insecticides and heavy metals. SlGSTE1 was significantly up-regulated by chlorpyrifos and xanthotoxin in the midgut of S. litura. The recombinant SlGSTE1 had Vmax (reaction rate of the enzyme saturated with the substrate) and Km (michaelis constant and equals to the substrate concentration at half of the maximum reaction rate of the enzyme) values of 27.95 ± 0.88 μmol/min/mg and 0.87 ± 0.028 mmol/L for glutathione, respectively, and Vmax and Km values of 22.96 ± 0.78 μmol/min/mg and 0.83 ± 0.106 mmol/L for 1-chloro-2,4-dinitrobenzene, respectively. In vitro enzyme indirect activity assay showed that the recombinant SlGSTE1 possessed high binding activities to the insecticides chlorpyrifos, deltamethrin, malathion, phoxim and dichloro-diphenyl-trichloroethane (DDT). SlGSTE1 showed higher binding activity to toxic heavy metals cadmium, chromium and lead than copper and zinc that are required for insect normal growth. Western blot analysis showed that SlGSTE1 was induced in the gut of larvae fed with chlorpyrifos or cadmium. SlGSTE1 also showed high peroxidase activity. All the results together indicate that SlGSTE1 may play an important role in the gut of S. litura to protect the insect from the toxic effects of these compounds and heavy metals. PMID:24863567

  6. Effect of nitrogen fertigation and sowing time on the expression of Cry2Ab and on mortality of Spodoptera litura in Bollgard II cotton.

    PubMed

    Saini, Manpreet Kaur; Dhawan, A K

    2014-03-01

    Toxin expression of Cry2Ab was studied in plant parts of Bollgard II cotton genotype MRC 7031 sown under different treatments of nitrogen application and planting dates. The expression was quantified by using Cry2Aa ELISA kit. Mean per cent mortality of one-day-old, 3rd and 5th instar larvae of Spodoptera litura was observed on different plant parts of MRC 7031 and their respective non-Bt cotton genotypes. The study deduced that mean maximum expression (19.24, 20.93 and 20.71 microg g(-1) in leaves, squares and bolls, respectively) of Cry2Ab was observed at higher nitrogen dose @ 300 kg ha(-1) (N3), while it was minimum (18.67, 20.44 and 20.14 microg g(-1) in leaves, squares and bolls, respectively) at low nitrogen dose @ 150 kg ha(-1) (N1). Studies conducted for different planting dates showed mean maximum expression (18.98, 20.72 and 20.42 microg g(-1) in leaves, squares and bolls, respectively) of Cry2Ab during late sown crop (15th May) as compared to early sown crop (15th April), the expression was 18.66, 20.32 and 20.06 microg g(-1) in leaves, squares and bolls, respectively. Quantitative expression of Cry2Ab was found to vary among different plant parts, i.e more in squares followed by bolls and leaves. Regarding mortality of different instars of S. litura, it was significantly more at higher nitrogen doses and it ranged from 83.04 to 96.27, 53.38 to 61.87 and 16.87 to 22.58% in case of S. litura one-day-old larvae, 3rd and 5th instar, respectively. While, non significant difference in mortalitywas observed during different sowing dates. PMID:24665755

  7. Integration of Insecticidal Protein Vip3Aa1 into Beauveria bassiana Enhances Fungal Virulence to Spodoptera litura Larvae by Cuticle and Per Os Infection▿

    PubMed Central

    Qin, Yi; Ying, Sheng-Hua; Chen, Ying; Shen, Zhi-Cheng; Feng, Ming-Guang

    2010-01-01

    The entomopathogenic fungus Beauveria bassiana acts slowly on insect pests through cuticle infection. Vegetative insecticidal proteins (Vip3A) of Bacillus thuringiensis kill lepidopteran pests rapidly, via per os infection, but their use for pest control is restricted to integration into transgenic plants. A transgenic B. bassiana strain (BbV28) expressing Vip3Aa1 (a Vip3A toxin) was thus created to infect the larvae of the oriental leafworm moth Spodoptera litura through conidial ingestion and cuticle adhesion. Vip3Aa1 (∼88 kDa) was highly expressed in the conidial cytoplasm of BbV28 and was detected as a digested form (∼62 kDa) in the larval midgut 18 and 36 h after conidial ingestion. The median lethal concentration (LC50) of BbV28 against the second-instar larvae feeding on cabbage leaves sprayed with conidial suspensions was 26.2-fold lower than that of the wild-type strain on day 3 and 1.1-fold lower on day 7. The same sprays applied to both larvae and leaves for their feeding reduced the LC50 of the transformant 17.2- and 1.3-fold on days 3 and 7, respectively. Median lethal times (LT50s) of BbV28 were shortened by 23 to 35%, declining with conidial concentrations. The larvae infected by ingestion of BbV28 conidia showed typical symptoms of Vip3A action, i.e., shrinkage and palsy. However, neither LC50 nor LT50 trends differed between BbV28 and its parental strain if the infection occurred through the cuticle only. Our findings indicate that fungal conidia can be used as vectors for spreading the highly insecticidal Vip3A protein for control of foliage feeders such as S. litura. PMID:20495052

  8. Target and non-target toxicity of botanical insecticide derived from Couroupita guianensis L. flower against generalist herbivore, Spodoptera litura Fab. and an earthworm, Eisenia foetida Savigny.

    PubMed

    Ponsankar, Athirstam; Vasantha-Srinivasan, Prabhakaran; Senthil-Nathan, Sengottayan; Thanigaivel, Annamalai; Edwin, Edward-Sam; Selin-Rani, Selvaraj; Kalaivani, Kandaswamy; Hunter, Wayne B; Alessandro, Rocco T; Abdel-Megeed, Ahmed; Paik, Chae-Hoon; Duraipandiyan, Veeramuthu; Al-Dhabi, Naif Abdullah

    2016-11-01

    Botanical insecticides may provide alternatives to synthetic insecticides for controlling Spodoptera litura (F.) and they are target specific, biodegradable, and harmless to mammals. Eight natural chemical compounds with larvicidal activity were identified from fraction F6 of C. guianensis flower extract. Probit analysis of 95% confidence level exposed an LC50 of 223ppm against S. litura third instar larvae. The growth and development of S. litura was affected in sub-lethal concentrations of fraction F6 (50, 100, 150 and 200ppm) compared to controls. Similarly nutritional indices values decreased significantly compared to controls. Fraction F6 also damaged the gut epithelial layer and brush border membrane (BBM). This study also resolved the effects of toxicity to non-target earthworm treated with fraction F6 and chemical pesticides (monotrophos and cypermethrin) and the results showed that fraction F6 had no harmful effect on E. fetida. Further, fraction F6 was eluted and sub fractions F6c (50ppm) showed high mortality against S. litura third instar larvae. Octacosane from fraction F6c was established and confirmed using IR spectrum and HPLC. The time of retention of fraction F6c was confirmed with the octacosane standard. Fraction F6 of C. guianensis extract caused dose-dependent mortality towards S. litura. Octacosane in fraction F6c was establish to be the prominent chemical compound associated with causing mortality but other compounds present in the fraction F6 were shown to be associated with changes in development of S. litura at low dosages. S. litura at low dosage. Therefore, these findings suggest that octacosane may be one of the major insecticidal compounds affecting S. litura survival. PMID:27476000

  9. Interaction of the Koinobiont Parasitoid Microplitis rufiventris of the Cotton Leafworm, Spodoptera littoralis, with Two Entomopathogenic Rhabditids, Heterorhabditis bacteriophora and Steinernema carpocapsae

    PubMed Central

    Atwa, Atwa A.; Hegazi, Esmat M.; Khafagi, Wedad E.; El-Aziz, Gehan M. Abd

    2013-01-01

    Entomopathogenic nematodes are generally considered beneficial organisms. However, they can affect beneficial insects such as parasitoids. The interaction between the entomopathogenic nematodes Heterorhabditis bacteriophora Poinar (Rhabditida: Heterorhabditidae) and Steinernema carpocapsae Weiser, and the parasitoid Microplitis rufiventris Kokujev (Hymenoptera: Braconidae) was investigated in the laboratory. In non-parasitized hosts, Spodoptera littoralis Boisduval (Lepidoptera: Noctuidae) larvae exposed to H. bacteriophora showed a higher percent mortality than those exposed to S. carpocapsae. Both nematodes were able to invade and propagate in non-parasitized S. littoralis larvae and those parasitized by M. rufiventris. Both nematode species reproduced in Microplitis-parasitized hosts, but there was a higher number of nematodes in non-parasitized larvae. S. carpocapsae yielded higher numbers of infective juveniles than H. bacteriophora. Generally, the number of nematodes harvested increased as their host's size increased. The interaction between the nematodes and parasitoid favored the nematodes when the nematodes were inoculated during the parasitoid egg stage or the young parasitoid larvae, thus giving the nematodes a better chance to grow and reproduce, resulting in the death of the parasitoid larvae. Conversely, when the nematodes were inoculated during the late larval instar of the parasitoid, the competition partially favored the wasp, thus giving approximately 50% of the wasps a better chance to develop, emerge, and reproduce, providing evidence that both nematodes and wasps could reproduce in the same host. Egg maturation of female wasps derived from nematode-infected hosts was not significantly different than those from control hosts. The combined application of nematodes and parasitoids may be beneficial if the detrimental effects of the nematodes on the parasitoid could be avoided by precisely timing the application strategies. It is clear that

  10. Effects of sublethal concentrations of the chitin synthesis inhibitor, hexaflumuron, on the development and hemolymph physiology of the cutworm, Spodoptera litura.

    PubMed

    Zhu, Qiqi; He, Yuan; Yao, Jing; Liu, Yinzhao; Tao, Liming; Huang, Qingchun

    2012-01-01

    The effects of sublethal concentrations 0.1, 0.5, and 1.2 µg mL(-1)of the chitin synthesis inhibitor, hexaflumuron, on larval growth and development, the count and proportion of hemocytes, and carbohydrate content (trehalose and glyceride) in hemolymph were investigated in the cutworm, Spodoptera litura (Fabricious) (Lepidoptera: Noctuidae). When 3(rd) instar larvae were subjected to the sublethal concentrations, there were dose-dependent effects on larval weight and length of each instar larvae, percent pupation and the duration of development. Most of the larvae died during the molting process at all concentrations. Few individuals from 0.5 and 1.2 µg mL(-1)concentrations could develop to the 6(th) instar, while the pupae emerging from the 0.1 µg mL(-1)concentrations did not exceed 16% of the number of the initial larvae. In 5(th)instar S. litura, the total number of hemocytes was significantly increased at 24 hours post-treatment, whereas the proliferation of hemocytes was inhibited, plasmatocyte pseudopodia contracted, and granulocyte expanded at 96 hours post-treatment. The increases of plasmatocyte count and the decreases of granulocyte count were dose-dependent. The longer treatment time of the sublethal concentrations increased the content of total carbohydrate and trehalose in hematoplasma, and was dose-dependent in hemocytes. The content of glyceride in hemolymph was significantly higher at 24 hours post-treatment, but gradually returned to normal levels at 96 hours post-treatment as compared with the control. The results suggested that sublethal concentrations of hexaflumuron reduced S. litura larval survival and interfered with hemolymph physiological balances. PMID:22958164

  11. Effects of Sublethal Concentrations of the Chitin Synthesis Inhibitor, Hexaflumuron, on the Development and Hemolymph Physiology of the Cutworm, Spodoptera litura

    PubMed Central

    Zhu, Qiqi; He, Yuan; Yao, Jing; Liu, Yinzhao; Tao, Liming; Huang, Qingchun

    2012-01-01

    The effects of sublethal concentrations 0.1, 0.5, and 1.2 µg mL-1of the chitin synthesis inhibitor, hexaflumuron, on larval growth and development, the count and proportion of hemocytes, and carbohydrate content (trehalose and glyceride) in hemolymph were investigated in the cutworm, Spodoptera litura (Fabricious) (Lepidoptera: Noctuidae). When 3rdinstar larvae were subjected to the sublethal concentrations, there were dose-dependent effects on larval weight and length of each instar larvae, percent pupation and the duration of development. Most of the larvae died during the molting process at all concentrations. Few individuals from 0.5 and 1.2 µg mL -1concentrations could develop to the 6thinstar, while the pupae emerging from the 0.1 µg mL -1concentrations did not exceed 16% of the number of the initial larvae. In 5thinstar S. litura, the total number of hemocytes was significantly increased at 24 hours post—treatment, whereas the proliferation of hemocytes was inhibited, plasmatocyte pseudopodia contracted, and granulocyte expanded at 96 hours post—treatment. The increases of plasmatocyte count and the decreases of granulocyte count were dose—dependent. The longer treatment time of the sublethal concentrations increased the content of total carbohydrate and trehalose in hematoplasma, and was dose—dependent in hemocytes. The content of glyceride in hemolymph was significantly higher at 24 hours post—treatment, but gradually returned to normal levels at 96 hours post—treatment as compared with the control. The results suggested that sublethal concentrations of hexaflumuron reduced S. litura larval survival and interfered with hemolymph physiological balances. PMID:22958164

  12. Very high-density lipoprotein and vitellin as carriers of novel biliverdins IXα with a farnesyl side-chain presumably derived from heme A in Spodoptera littoralis.

    PubMed

    Kayser, Hartmut; Nimtz, Manfred; Ringler, Philippe; Müller, Shirley A

    2016-01-01

    Bilins in complex with specific proteins play key roles in many forms of life. Biliproteins have also been isolated from insects; however, structural details are rare and possible functions largely unknown. Recently, we identified a high-molecular weight biliprotein from a moth, Cerura vinula, as an arylphorin-type hexameric storage protein linked to a novel farnesyl biliverdin IXα; its unusual structure suggests formation by cleavage of mitochondrial heme A. In the present study of another moth, Spodoptera littoralis, we isolated two different biliproteins. These proteins were identified as a very high-density lipoprotein (VHDL) and as vitellin, respectively, by mass spectrometric sequencing. Both proteins are associated with three different farnesyl biliverdins IXα: the one bilin isolated from C. vinula and two new structurally closely related bilins, supposed to be intermediates of heme A degradation. The different bilin composition of the two biliproteins suggests that the presumed oxidations at the farnesyl side-chain take place mainly during egg development. The egg bilins are supposedly transferred from hemolymph VHDL to vitellin in the female. Both biliproteins show strong induced circular dichroism activity compatible with a predominance of the M-conformation of the bilins. This conformation is opposite to that of the arylphorin-type biliprotein from C. vinula. Electron microscopy of the VHDL-type biliprotein from S. littoralis provided a preliminary view of its structure as a homodimer and confirmed the biochemically determined molecular mass of ∼350 kDa. Further, images of S. littoralis hexamerins revealed a 2 × 3 construction identical to that known from the hexamerin from C. vinula. PMID:26546815

  13. Parasitization of fall armyworm by the eulophid parasitoid Euplectrus platyhypenae

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fall armyworm, Spodoptera frugiperda (J. E. Smith), is attacked by parasitoids from several different families of Hymenoptera and Diptera. One species, Euplectrus platyhypenae Howard (Hymenoptera: Eulophidae), has been consistently recovered from larvae in whorl-stage corn and from larvae feeding i...

  14. Antibiosis among selected paspalum taxa to the fall armyworm (Lepidoptera: Noctuidae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Twenty six accessions of the warm-season perennial grass, Paspalum spp., were evaluated for response to the fall armyworm Spodoptera frugiperda (J. E. Smith), an important pest of turfgrass. In growth chamber, excised clipping studies, P. vaginatum 03-539-31 and P. vaginatum 03-525-22 were the most ...

  15. Physiological and biochemical bases of fall armyworm resistance in the seedlings of maize inbred lines

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Four CIMMYT maize inbred lines (i.e., CML333, CML335, CML 336, and CML338), and a susceptible (i.e., AB24E) and resistant (i.e., Mp780) control were examined for the mechanisms of fall armyworm (FAW), Spodoptera frugiperda (JE Smith) (Lepidoptera: Noctuidae) resistance. The six inbred lines were ev...

  16. Identification of fall armyworm (Lepidoptera:Noctuidae) host strains using male-derived spermatophores

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Laboratory experiments were designed to identify the host strain paternity of fall armyworm [Spodoptera frugiperda (J. E. Smith)] mated females. In no-choice tests, corn or rice strain females were placed in cages with males of the opposite strain. After 48 h, females were dissected and spermatoph...

  17. Attraction of fall armyworm males (Lepidoptera: Noctuidae) to host strain females

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Attraction of wild male fall armyworm, Spodoptera frugiperda (J. E. Smith), was compared in trapping experiments during 2005 – 2009 in Florida. Traps were baited either with a commercial sex pheromone lure or corn and rice strain females obtained from laboratory colonies. Over 7400 male moths were...

  18. Field screening of sweet sorghum inbred lines for pest resistance and biomass production

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Sweet sorghum [Sorghum bicolor (L.) Moench] is one of the favorable biofuel feedstocks for ethanol production. Fall armyworm [Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae)] is one of the most serious foliar-feeding insect pests in sorghum production in the southeastern US states. Sev...

  19. Effect of eastern gamagrass on fall armyworm and corn earworm development

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The fall armyworm, Spodoptera frugiperda (J.E. Smith) and the corn earworm, Helicoverpa zea (Boddie) are two important corn pests in the southern U.S. states. Effect of the leaves from the corn relative, the Eastern gamagrass (Tripsacum dactyloides L.) on fall armyworm and corn earworm development ...

  20. Sex pheromones of the southern armyworm moth: isolation, identification, and synthesis.

    PubMed

    Jacobson, M; Redfern, R E; Jones, W A; Aldridge, M H

    1970-10-30

    Two sex pheromones have been isolated from the female southern armyworm moth, Prodenia eridania (Cramer), and identified as cis-9-tetradecen-1-ol acetate, identical with the sex pheromone of the fall armyworm moth, Spodoptera frugiperda (J. E. Smith), and cis-9,trans-12-tetradecadien-1-ol acetate. PMID:5507205

  1. Genetic variation in neonate behavior of fall armyworm (Lepidoptera:Noctuidae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bioassays were developed to test plant selection of fall armyworm [Spodoptera frugiperda (J. E. Smith)] host strains to corn and stargrass. Neonate larvae from 3 corn strain and 3 rice strain colonies preferentially selected corn sections over stargrass sections in petri dish choice tests. However...

  2. Modeling seasonal migration of fall armyworm moths

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fall armyworm, Spodoptera frugiperda (J. E. Smith), is a highly mobile insect pest of a wide range of host crops. However, this pest of tropical origin cannot survive extended periods of freezing temperature, but must repeat a series of northward migratory flights each spring if it is to re-infest ...

  3. Inferring the annual migration patterns of fall armyworm (Lepidoptera: Noctuidae) in the U.S. from mitochondrial haplotypes.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Spodoptera frugiperda (J.E. Smith) or fall armyworm is an important agricultural pest of a number of crops in the Western Hemisphere. In the U.S., infestations in corn acreages extend from the Mexican to the Canadian border. Because fall armyworm does not survive prolonged ground freezing the infe...

  4. Evaluation of whorl damage by fall armyworm (Lepidoptera:Noctuidae) on field and greenhouse grown sweet sorghum plants

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The fall armyworm [Spodoptera frugiperda (Lepidoptera: Noctuidae)] is an economically important pest of sorghum [Sorghum bicolor (L) Moench]. However, resistance to fall armyworm in sweet sorghum has not been extensively studied. A collection of primarily sweet sorghum accessions were evaluated in t...

  5. Demonstration using field collections that Argentina fall armyworm populations exhibit strain-specific host plant preference

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Spodoptera frugiperda, the fall armyworm, is a major economic pest throughout the Western Hemisphere of corn (maize), cotton, sorghum, and a variety of agricultural grasses and vegetable crops. Studies in the United States, the Caribbean, and Brazil demonstrated the existence of two subpopulations ...

  6. Development and feeding of fall armyworm, on Miscanthus x giganteus and switchgrass

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Observations of fall armyworm [Spodoptera frugiperda (J.E. Smith)] larvae infesting plots of Miscanthus x giganteus Greef and Deuter ex Hodkinson and Renvoize prompted laboratory-based tests of survival, development and feeding preferences on leaf tissue from M. x giganteus and switchgrass (Panicum ...

  7. Parasitoids attacking fall armyworm (Lepidoptera: Noctuidae) in sweet corn habitats

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fall armyworm larvae, Spodoptera frugiperda (J. E. Smith), were collected from sweet corn plants (Zea mays L.) in fields located in three south Florida counties. Fields were sampled from 2010 – 2015 during the fall and spring seasons. Larvae were brought back to the laboratory to complete developme...

  8. Sunn hemp as a ground cover to manage fall armyworm populations

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fall armyworm (FAW), Spodoptera frugiperda, is a serious pest of sweet corn in south Florida and a pest of other vegetable, row, and forage crops in the southeastern, mid-Atlantic, and central U.S. It is a migratory pest, moving north each season from overwintering areas in southern Texas and south...

  9. Efficacy of silk channel injections with insecticides for management of Lepidopteran pests of sweet corn

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The primary Lepidopteran pests of sweet corn in Georgia are the corn earworm, Helicoverpa zea (Boddie), and the fall armyworm, Spodoptera frugiperda (J.E. Smith). Control of these pests typically requires multiple insecticide applications from first silking until harvest, with commercial growers fre...

  10. Expression of a synthetic Bacillus thuringiensis endotoxin to enhance resistance against fall armyworm in bahiagrass

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bahiagrass is a low input, drought tolerant and disease resistant warm season turfgrass used for forage and turf in the southeastern U.S. and other subtropical regions of the world. Fall armyworm, Spodoptera frugiperda (J. E. Smith) is a destructive migratory pest of many tropical and subtropical g...

  11. Feeding deterrence and inhibitory effects of bee balm (Monarda didyma) leaves on fall armyworm

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The fall armyworm [Spodoptera frugiperda (J.E. Smith)] is a serious pest of many field and horticulture crops. Because of the many advantages for the use of plant-derived pesticides, we tested whether bee balm (Monarda didyma L.) leaves could have feeding deterrence on fall armyworm. When S. frugipe...

  12. Impact of transgenic sweet corn silks to two noctuid pests

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Eight Bacillus thuringiensis (Bt) transgenic sweet corn hybrids were evaluated (with two controls) for their efficacy against two ear-feeding insects; the corn earworm [Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae)], and the fall armyworm (Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuid...

  13. Fall Armyworm susceptibility to Bollgard I, Bollgard II, and Widestrike cotton as determined by a leaf-dish assay.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Fall armyworm Spodoptera frugiperda (J.E. Smith), has received little attention as boll weevil eradication progresses into the second full season in the Rio Grande Valley of South Texas. Cotton varieties containing the endotoxins from Bacillus thuringiensis (Cry1Ac = Bollgard®, Cry1Ac + Cry1Ab ...

  14. Evaluations of Bollgard, Bollgard II, and Widestrike technologies against beet and fall armyworm larvae (Lepidoptera: Noctuidae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Transgenic cottons containing the Bollgard®, Bollgard II® and WideStrike® traits were grown in 2005 and 2007 to examine the efficacy against beet [Spodoptera exigua (Hübner)] and fall armyworms [S. frugiperda (J. E. Smith)]. Results suggest that both dual-gene traits are more efficacious against th...

  15. Using haplotypes to monitor the migration of fall armyworm (Lepidoptera: Noctuidae) corn-strain populations from Texas and Florida

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Spodoptera frugiperda (J.E. Smith; Lepidoptera: Noctuidae) or fall armyworm is an important agricultural pest of a number of crops in the western hemisphere. Two morphologically identical host strains of fall armyworm exist, the rice-strain and corn-strain, with the latter inflicting substantial eco...

  16. Cowpea chloroplastic ATP synthase is the source of multiple plant defense elicitors during insect herbivory

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Plant responses to damage vary dependant upon the nature of the biotic and abiotic stresses. We recently described an elicitor, from Fall armyworm (Spodoptera frugiperda) oral secretions (OS) termed inceptin, derived from chloroplastic ATP synthase '-subunit (cATPC) proteins that activate phytohormo...

  17. Review of fall armyworm (Lepidoptera: Noctuidae) genetic complexity and migration

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The fall armyworm, Spodoptera frugiperda (J. E. Smith) is a significant economic pest in the western hemisphere, causing substantial losses in corn, sorghum, forage and turf grasses (Luginbill 1928, Sparks 1979). Although fall armyworm does not survive severe winters, it infests most of the central...

  18. Using stable isotope analysis to examine the fall armyworm (Lepidoptera: Noctuidae) host strains in a cotton habitat

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Spodoptera frugiperda (J.E. Smith; Lepidoptera: Noctuidae) or fall armyworm is an important agricultural pest of a number of crops in the western hemisphere, including cotton. Two genetically distinct but morphologically identical strains (R-strain and C-strain) exist that differ physiologically and...

  19. Damage and survivorship of fall armyworm (Lepidoptera: noctuidae) on transgenic field corn expressing Bacillus thuringiensis cry proteins

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Field corn, Zea mays L. plants expressing Cry1Ab and Cry1F insecticidal crystal (Cry) proteins of Bacillus thuringiensis (Bt) Berliner are planted on considerable acreage across the Southern region of the United States. The fall armyworm, Spodoptera frugiperda (J. E. Smith), is an economically impo...

  20. Fall armyworm (Lepidoptera: Noctuidae) development survivorship and damage on cotton plants expressing insecticidal plant-incorporated protectants

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Cotton, Gossypium hirsutum (L.), plants expressing insecticidal crystal (Cry) proteins of Bacillus thuringiensis (Bt) Berliner are planted on significant acreage across the Southern region of the United States. Fall armyworm, Spodoptera frugiperda (J. E. Smith), can be a significant cotton pest in ...

  1. Expression of crylFa in bahiagrass enhances resistance to fall armyworm

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Bahiagrass (Paspalum notatum var. Flugge) is the predominant forage grass in Florida and in subtropical regions worldwide. To improve pest resistance against fall armyworm (Spodoptera frugiperda J. E. Smith), an optimized cry1Fa gene encoding a '-endotoxin from Bacillus thuringiensis was synthesized...

  2. Supplemental control of lepidopterous pests on BT transgenic sweet corn with biologically based spray treatments

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Biologically based spray treatments, including nucleopolyhedroviruses, neem, and spinosad, were evaluated as supplemental controls for the fall armyworm, Spodoptera frugiperda (J. E. Smith), and corn earworm, Helicoverpa zea (Boddie), on transgenic sweet corn, Zea mays (L.), expressing a Cry1Ab toxi...

  3. Survey for hymenopteran and dipteran parasitoids of the fall armyworm (Lepidoptera: Noctuidae) in Chiapas, Mexico

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A survey of hymenopteran and dipteran parasitoids of the fall armyworm (FAW), Spodoptera frugiperda (J. E. Smith) larvae was conducted to determine their occurrence and parasitism rates in Western Chiapas, Mexico. FAW larvae were collected from whorl-stage cornfields in Chiapas in the region called ...

  4. Spatial patterns of aflatoxin levels in relation to ear-feeding insect damage in pre-harvest corn

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Key impediments to increased corn yield and quality in the southeastern US coastal plain region are damage by ear-feeding insects and aflatoxin contamination caused by infection of Aspergillus flavus. Key ear-feeding insects are corn earworm, Helicoverpa zea, fall armyworm, Spodoptera frugiperda, m...

  5. Transgenic cotton age effects on lepidopteran larvae mortality

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Leaves from plants containing various transgenic traits (Non-Bt, Bollgard, Bollgard II, and WideStrike)were assayed for bioactivity in the laboratory against bollworm, Helicoverpa zea (Boddie), beet armyworm, Spodoptera frugiperda (J. E. Smith), and cabbage looper, Trichoplusia ni (Hubner). About 5...

  6. Impact of Fall Armyworm Survival in Bt Crops on Survival and Damage Potential of Subsequent Generations

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fall armyworm, Spodoptera frugiperda (J. E. Smith), colonies were compared to determine whether development on Bt corn or Bt cotton impacted survival and damage potential of subsequent generations on Bt or non-Bt cotton. Late instars of fall armyworm were collected from Bt and non-Bt sweet corn to ...

  7. Susceptibility of field populations of the fall armyworm (Lepidoptera: Noctuidae) from Florida and Puerto Rico to purified Cry1F protein and corn leaf tissue containing single and pyramided Bt genes

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Larval survival of Cry1F-susceptible (FL), -resistant (PR and Cry1F-RR), and -heterozygous (FL x PR and Cry1F-RS) populations of the fall armyworm, Spodoptera frugiperda (J.E. Smith) to purified Cry1F protein and corn leaf tissue of seven Bacillus thuringiensis (Bt) corn hybrids and five non-Bt corn...

  8. Laboratory evaluations of Lepidopteran-active soybean seed treatments on survivorship of fall armyworm (Lepidoptera:Noctuidae) larvae

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Two anthranilic diamide insecticides, chlorantraniliprole and cyantraniliprole, were evaluated as soybean, Glycine max L., seed treatments for control of fall armyworm, Spodoptera frugiperda (J. E. Smith). Bioassays were conducted using 2nd instar larvae and plants from both field and greenhouse gr...

  9. Genetics of migratory and invasive pests in Florida

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Overview of genetic research on migratory and invasive pests being performed at CMAVE. Emphasis will be on efforts to monitor the movements of fall armyworm, Spodoptera frugiperda (J.E. Smith)(Lepidoptera: Noctuidae) during its annual migration, model the behavior in order to predict the effect of r...

  10. Studies on fall armyworm migration and monitoring

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Spodoptera frugiperda (J.E. Smith; Lepidoptera: Noctuidae) or fall armyworm is an important agricultural pest of a number of crops in thewestern hemisphere. Two morphologically identical host strains of fall armyworm exist, the rice-strain and corn-strain, with the latter inflicting substantial eco...

  11. Texas is the overwintering source of fall armyworm in central Pennsylvainia: implications for migration into the northeastern United States

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fall armyworm, Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae), infestations in most of North America arise from annual migrations of populations that overwinter in southern Texas and Florida. Cytochrome Oxidase I haplotype profiles within the fall armyworm corn-strain, the subgroup tha...

  12. Fall Armyworm in the Southeast

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Two separate experiments testing fall armyworm (Spodoptera frugiperda) migration patterns were set up in the southeastern U.S. in 2012. Previous results showed that moths from progeny of overwintering populations from south Texas were found west of the Chattahoochee-Flint-Apalachicola river basin, ...

  13. Use of benzimidazole agar plates to assess fall armyworm (Lepidoptera: Noctuidae) feeding on excised maize and sorghum leaves

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The fall armyworm, Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) is an economically significant pest of sorghum and maize. To screen sorghum and maize germplasm for resistance to fall armyworm feeding, field, greenhouse, or lab bioassays are often utilized individually or in combinatio...

  14. Fall armyworm migration patterns.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fall armyworm, Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae), infestations in most of North America arise from annual migrations of populations that overwinter in southern Texas and Florida. Cytochrome Oxidase I haplotype profiles within the fall armyworm corn-strain, the subgroup tha...

  15. Physiological, nutritional and biochemical bases of corn resistance to foliage-feeding fall armyworm (Lepidoptera:noctuidae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Three corn (Zea mays) germplasm lines, Ab24E (susceptible control), Mp708 (resistant control), and a locally selected partial inbred line FAW7050 (resistant), were examined for Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) resistance. Nutritional [i.e., total protein, amino acids, gl...

  16. Identifying resistance in corn to southwestern corn borer, fall armyworm, and corn earworm

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Southwestern corn borer, Diatraea grandiosella Dyar; fall armyworm, Spodoptera frugiperda (J.E. Smith); and corn earworm, Helicoverpa zea Boddie, are major insect pests of corn, Zea mays L., in the southern United States. Corn germplasm with resistance to leaf feeding by southwestern corn borer a...

  17. Genetic mapping of fall armyworm resistance in Zoysiagrass

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Molecular tools have not identified markers for host plant resistance to fall armyworm [Spodoptera frugiperda (J. E. Smith)], one of the most damaging insect pests in the southeastern U.S., in turfgrasses. Available QTLs in corn have not been further assessed for utility as comparative markers for ...

  18. Using intron sequence comparisons in the triose-phosphate isomerase gene to study the divergence of the fall armyworm host strains

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The Noctuid moth, Spodoptera frugiperda (the fall armyworm), is endemic to the Western Hemisphere and appears to be undergoing sympatric speciation to produce two subpopulations that differ in their choice of host plants. The diverging “rice strain” and “corn strain” are morphologically indistinguis...

  19. Portable exhauster POR-007/Skid E and POR-008/Skid F storage plan

    SciTech Connect

    Nelson, O.D.

    1998-07-25

    This document provides storage requirements for 1,000 CFM portable exhausters POR-O07/Skid E and POR-008/Skid F. These requirements are presented in three parts: preparation for storage, storage maintenance and testing, and retrieval from storage. The exhauster component identification numbers listed in this document contain the prefix POR-007 or POR-008 depending on which exhauster is being used.

  20. SeGSTo, a novel glutathione S-transferase from the beet armyworm (Spodoptera exigua), involved in detoxification and oxidative stress.

    PubMed

    Xu, Pengfei; Han, Ningning; Kang, Tinghao; Zhan, Sha; Lee, Kwang Sik; Jin, Byung Rae; Li, Jianhong; Wan, Hu

    2016-09-01

    Members of the glutathione S-transferase superfamily can protect organisms against oxidative stress. In this study, we characterized an omega glutathione S-transferase from Spodoptera exigua (SeGSTo). The SeGSTo gene contains an open reading frame (ORF) of 744 nucleotides encoding a 248-amino acid polypeptide. The predicted molecular mass and isoelectric point of SeGSTo are 29007 Da and 7.74, respectively. Multiple amino acid sequence alignment analysis shows that the SeGSTo sequence is closely related to the class 4 GSTo of Bombyx mori BmGSTo4 (77 % protein sequence similarity). Homologous modeling and molecular docking reveal that Cys35 may play an essential role in the catalytic process. Additionally, the phylogenetic tree indicates that SeGSTo belongs to the omega group of the GST superfamily. During S. exigua development, SeGSTo is expressed in the midgut of the fifth instar larval stage, but not in the epidermis or fat body. Identification of recombinant SeGSTo via SDS-PAGE and Western blot shows that its molecular mass is 30 kDa. The recombinant SeGSTo was able to protect super-coiled DNA from damage in a metal-catalyzed oxidation (MCO) system and catalyze the 1-chloro-2,4-dinitrobenzene (CDNB), but not 1,2-dichloro-4-nitrobenzene (DCNB), 4-nitrophenethyl bromide (4-NPB), or 4-nitrobenzyl chloride (4-NBC). The optimal reaction pH and temperature were 8 and 50 °C, respectively, in the catalysis of CDNB by recombinant SeGSTo. The mRNA expression of SeGSTo was up-regulated by various oxidative stresses, such as CdCl2, CuSO4, and isoprocarb, and the catalytic activity of recombinant SeGSTo was noticeably inhibited by heavy metals (Cu(2+) and Cd(2+)) and various pesticides. Taken together, these results indicate that SeGSTo plays an important role in the antioxidation and detoxification of pesticides. PMID:27230212

  1. Two different Bacillus thuringiensis toxin genes confer resistance to beet armyworm (Spodoptera exigua Hübner) in transgenic Bt-shallots (Allium cepa L.).

    PubMed

    Zheng, Si-Jun; Henken, Betty; de Maagd, Ruud A; Purwito, Agus; Krens, Frans A; Kik, Chris

    2005-06-01

    Agrobacterium-mediated genetic transformation was applied to produce beet armyworm (Spodoptera exigua Hübner) resistant tropical shallots (Allium cepa L. group Aggregatum). A cry1Ca or a H04 hybrid gene from Bacillus thuringiensis, driven by the chrysanthemum ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit (Rubisco SSU) promoter, along with the hygromycin phosphotransferase gene (hpt) driven by the CaMV 35S promoter, was employed for genetic transformation. An average transformation frequency of 3.68% was obtained from two shallot cultivars, Tropix and Kuning. After transfer of the in vitro plants to the greenhouse 69% of the cry1Ca and 39% of the H04 transgenic shallots survived the first half year. After one year of cultivation in the greenhouse the remaining cry1Ca and H04 transgenic plants grew vigorously and had a normal bulb formation, although the cry1Ca transgenic plants (and controls) had darker green leaves compared to their H04 counterparts. Standard PCR, adaptor ligation PCR and Southern analyses confirmed the integration of T-DNA into the shallot genome. Northern blot and ELISA analyses revealed expression of the cry1Ca or H04 gene in the transgenic plants. The amount of Cry1Ca expressed in transgenic plants was higher than the expression levels of H04 (0.39 vs. 0.16% of the total soluble leaf proteins, respectively). There was a good correlation between protein expression and beet armyworm resistance. Cry1Ca or H04 gene expression of at least 0.22 or 0.08% of the total soluble protein in shallot leaves was sufficient to give a complete resistance against beet armyworm. This confirms earlier observations that the H04 toxin is more toxic to S. exigua than the Cry1Ca toxin. The results from this study suggest that the cry1Ca and H04 transgenic shallots developed could be used for introducing resistance to beet armyworm in (sub) tropical shallot. PMID:16145834

  2. Identification of Metabolically Active Bacteria in the Gut of the Generalist Spodoptera littoralis via DNA Stable Isotope Probing Using 13C-Glucose

    PubMed Central

    Boland, Wilhelm

    2013-01-01

    Guts of most insects are inhabited by complex communities of symbiotic nonpathogenic bacteria. Within such microbial communities it is possible to identify commensal or mutualistic bacteria species. The latter ones, have been observed to serve multiple functions to the insect, i.e. helping in insect reproduction1, boosting the immune response2, pheromone production3, as well as nutrition, including the synthesis of essential amino acids4, among others.     Due to the importance of these associations, many efforts have been made to characterize the communities down to the individual members. However, most of these efforts were either based on cultivation methods or relied on the generation of 16S rRNA gene fragments which were sequenced for final identification. Unfortunately, these approaches only identified the bacterial species present in the gut and provided no information on the metabolic activity of the microorganisms. To characterize the metabolically active bacterial species in the gut of an insect, we used stable isotope probing (SIP) in vivo employing 13C-glucose as a universal substrate. This is a promising culture-free technique that allows the linkage of microbial phylogenies to their particular metabolic activity. This is possible by tracking stable, isotope labeled atoms from substrates into microbial biomarkers, such as DNA and RNA5. The incorporation of 13C isotopes into DNA increases the density of the labeled DNA compared to the unlabeled (12C) one. In the end, the 13C-labeled DNA or RNA is separated by density-gradient ultracentrifugation from the 12C-unlabeled similar one6. Subsequent molecular analysis of the separated nucleic acid isotopomers provides the connection between metabolic activity and identity of the species. Here, we present the protocol used to characterize the metabolically active bacteria in the gut of a generalist insect (our model system), Spodoptera littoralis (Lepidoptera, Noctuidae). The phylogenetic analysis of the DNA

  3. Secretions from the ventral eversible gland of Spodoptera exigua caterpillars activate defense-related genes and induce emission of volatile organic compounds in tomato, Solanum lycopersicum

    PubMed Central

    2014-01-01

    Background Plant induced defense against herbivory are generally associated with metabolic costs that result in the allocation of photosynthates from growth and reproduction to the synthesis of defense compounds. Therefore, it is essential that plants are capable of sensing and differentiating mechanical injury from herbivore injury. Studies have shown that oral secretions (OS) from caterpillars contain elicitors of induced plant responses. However, studies that shows whether these elicitors originated from salivary glands or from other organs associated with feeding, such as the ventral eversible gland (VEG) are limited. Here, we tested the hypothesis that the secretions from the VEG gland of Spodoptera exigua caterpillars contain elicitors that induce plant defenses by regulating the expression of genes involved in the biosynthesis of volatile organic compounds (VOCs) and other defense-related genes. To test this hypothesis, we quantified and compared the activity of defense-related enzymes, transcript levels of defense-related genes and VOC emission in tomato plants damaged by S. exigua caterpillars with the VEG intact (VEGI) versus plants damaged by caterpillars with the VEG ablated (VEGA). Results The quantified defense-related enzymes (i.e. peroxidase, polyphenol oxidase, and lipoxigenase) were expressed in significantly higher amounts in plants damaged by VEGI caterpillars than in plants damaged by VEGA caterpillars. Similarly, the genes that encode for the key enzymes involved in the biosynthesis of jasmonic acid and terpene synthase genes that regulate production of terpene VOCs, were up-regulated in plants damaged by VEGI caterpillars. Moreover, the OS of VEGA caterpillars were less active in inducing the expression of defense genes in tomato plants. Increased emissions of VOCs were detected in the headspace of plants damaged by VEGI caterpillars compared to plants damaged by VEGA caterpillars. Conclusion These results suggest that the VEG of S. exigua

  4. Consequences of POR mutations and polymorphisms.

    PubMed

    Miller, Walter L; Agrawal, Vishal; Sandee, Duanpen; Tee, Meng Kian; Huang, Ningwu; Choi, Ji Ha; Morrissey, Kari; Giacomini, Kathleen M

    2011-04-10

    P450 oxidoreductase (POR) transports electrons from NADPH to all microsomal cytochrome P450 enzymes, including steroidogenic P450c17, P450c21 and P450aro. Severe POR mutations A287P (in Europeans) and R457H (in Japanese) cause the Antley-Bixler skeletal malformation syndrome (ABS) plus impaired steroidogenesis (causing genital anomalies), but the basis of ABS is unclear. We have characterized the activities of ∼40 POR variants, showing that assays based on P450c17 activities, but not cytochrome c assays, correlate with the clinical phenotype. The human POR gene is highly polymorphic: the A503V sequence variant, which decreases P450c17 activities to ∼60%, is found on ∼28% of human alleles. A promoter polymorphism (∼8% of Asians and ∼13% of Caucasians) at -152 reduces transcriptional activity by half. Screening of 35 POR variants showed that most mutants lacking activity with P450c17 or cytochrome c also lacked activity to support CYP1A2 and CYP2C19 metabolism of EOMCC (a fluorogenic non-drug substrate), although there were some remarkable differences: Q153R causes ABS and has ∼30% of wild-type activity with P450c17 but had 144% of WT activity with CYP1A2 and 284% with CYP2C19. The effects of POR variants on CYP3A4, which metabolizes nearly 50% of clinically used drugs, was examined with multiple, clinically relevant drug substrates, showing that A287P and R457H dramatically reduce drug metabolism, and that A503V variably impairs drug metabolism. The degree of activity can vary with the drug substrate assayed, as the drugs can influence the conformation of the P450. POR is probably an important contributor to genetic variation in both steroidogenesis and drug metabolism. PMID:21070833

  5. Consequences of POR mutations and polymorphisms

    PubMed Central

    Miller, Walter L.; Agrawal, Vishal; Sandee, Duanpen; Tee, Meng Kian; Huang, Ningwu; Choi, Ji Ha; Morrissey, Kari; Giacomini, Kathleen M.

    2015-01-01

    P450 oxidoreductase (POR) transports electrons from NADPH to all microsomal cytochrome P450 enzymes, including steroidogenic P450c17, P450c21 and P450aro. Severe POR mutations A287P (in Europeans) and R457H (in Japanese) cause the Antley-Bixler skeletal malformation syndrome (ABS) plus impaired steroidogenesis (causing genital anomalies), but the basis of ABS is unclear. We have characterized the activities of ~40 POR variants, showing that assays based on P450c17 activities, but not cytochrome c assays, correlate with the clinical phenotype. The human POR gene is highly polymorphic: the A503V sequence variant, which decreases P450c17 activities to ~60%, is found on ~28% of human alleles. A promoter polymorphism (~8% of Asians and ~13% of Caucasians) at −152 reduces transcriptional activity by half. Screening of 35 POR variants showed that most mutants lacking activity with P450c17 or cytochrome c also lacked activity to support CYP1A2 and CYP2C19 metabolism of EOMCC (a fluorogenic non-drug substrate), although there were some remarkable differences: Q153R causes ABS and has ~30% of wild-type activity with P450c17 but had 144% of WT activity with CYP1A2 and 284% with CYP2C19. The effects of POR variants on CYP3A4, which metabolizes nearly 50% of clinically used drugs, was examined with multiple, clinically-relevant drug substrates, showing that A287P and R457H dramatically reduce drug metabolism, and that A503V variably impairs drug metabolism. The degree of activity can vary with the drug substrate assayed, as the drugs can influence the conformation of the P450. POR is probably an important contributor to genetic variation in both steroidogenesis and drug metabolism. PMID:21070833

  6. Desigualdades por cáncer

    Cancer.gov

    Información básica de las desigualdades en salud por cáncer en EE. UU., factores que contribuyen a la carga desproporcionada del cáncer en algunos grupos y ejemplos de desigualdades en incidencia y mortalidad entre ciertos grupos de la población.

  7. Effects of juvenile hormone (JH) analog insecticides on larval development and JH esterase activity in two spodopterans.

    PubMed

    El-Sheikh, El-Sayed A; Kamita, Shizuo G; Hammock, Bruce D

    2016-03-01

    Juvenile hormone analog (JHA) insecticides are biological and structural mimics of JH, a key insect developmental hormone. Toxic and anti-developmental effects of the JHA insecticides methoprene, fenoxycarb, and pyriproxyfen were investigated on the larval and pupal stages of Spodoptera littoralis and Spodoptera frugiperda. Bioassays showed that fenoxycarb has the highest toxicity and fastest speed of kill in 2nd instar S. littoralis. All three JHAs affected the development of 6th instar (i.e., final instar) and pupal S. frugiperda. JH esterase (JHE) is a critical enzyme that helps to regulate JH levels during insect development. JHE activity in the last instar S. littoralis and S. frugiperda was 11 and 23 nmol min(-1) ml(-1) hemolymph, respectively. Methoprene and pyriproxyfen showed poor inhibition of JHE activity from these insects, whereas fenoxycarb showed stronger inhibition. The inhibitory activity of fenoxycarb, however, was more than 1000-fold lower than that of OTFP, a highly potent inhibitor of JHEs. Surprisingly, topical application of methoprene, fenoxycarb or pyriproxyfen on 6th instars of S. littoralis and S. frugiperda prevented the dramatic reduction in JHE activity that was found in control insects. Our findings suggest that JHAs may function as JH agonists that play a disruptive role or a hormonal replacement role in S. littoralis and S. frugiperda. PMID:26969437

  8. Structural Insights into the PorK and PorN Components of the Porphyromonas gingivalis Type IX Secretion System

    PubMed Central

    Gorasia, Dhana G.; Veith, Paul D.; Hanssen, Eric G.; Glew, Michelle D.; Sato, Keiko; Yukitake, Hideharu; Nakayama, Koji; Reynolds, Eric C.

    2016-01-01

    The type IX secretion system (T9SS) has been recently discovered and is specific to Bacteroidetes species. Porphyromonas gingivalis, a keystone pathogen for periodontitis, utilizes the T9SS to transport many proteins including the gingipain virulence factors across the outer membrane and attach them to the cell surface via a sortase-like mechanism. At least 11 proteins have been identified as components of the T9SS including PorK, PorL, PorM, PorN and PorP, however the precise roles of most of these proteins have not been elucidated and the structural organization of these components is unknown. In this study, we purified PorK and PorN complexes from P. gingivalis and using electron microscopy we have shown that PorN and the PorK lipoprotein interact to form a 50 nm diameter ring-shaped structure containing approximately 32–36 subunits of each protein. The formation of these rings was dependent on both PorK and PorN, but was independent of PorL, PorM and PorP. PorL and PorM were found to form a separate stable complex. PorK and PorN were protected from proteinase K cleavage when present in undisrupted cells, but were rapidly degraded when the cells were lysed, which together with bioinformatic analyses suggests that these proteins are exposed in the periplasm and anchored to the outer membrane via the PorK lipid. Chemical cross-linking and mass spectrometry analyses confirmed the interaction between PorK and PorN and further revealed that they interact with the PG0189 outer membrane protein. Furthermore, we established that PorN was required for the stable expression of PorK, PorL and PorM. Collectively, these results suggest that the ring-shaped PorK/N complex may form part of the secretion channel of the T9SS. This is the first report showing the structural organization of any T9SS component. PMID:27509186

  9. Structural Insights into the PorK and PorN Components of the Porphyromonas gingivalis Type IX Secretion System.

    PubMed

    Gorasia, Dhana G; Veith, Paul D; Hanssen, Eric G; Glew, Michelle D; Sato, Keiko; Yukitake, Hideharu; Nakayama, Koji; Reynolds, Eric C

    2016-08-01

    The type IX secretion system (T9SS) has been recently discovered and is specific to Bacteroidetes species. Porphyromonas gingivalis, a keystone pathogen for periodontitis, utilizes the T9SS to transport many proteins including the gingipain virulence factors across the outer membrane and attach them to the cell surface via a sortase-like mechanism. At least 11 proteins have been identified as components of the T9SS including PorK, PorL, PorM, PorN and PorP, however the precise roles of most of these proteins have not been elucidated and the structural organization of these components is unknown. In this study, we purified PorK and PorN complexes from P. gingivalis and using electron microscopy we have shown that PorN and the PorK lipoprotein interact to form a 50 nm diameter ring-shaped structure containing approximately 32-36 subunits of each protein. The formation of these rings was dependent on both PorK and PorN, but was independent of PorL, PorM and PorP. PorL and PorM were found to form a separate stable complex. PorK and PorN were protected from proteinase K cleavage when present in undisrupted cells, but were rapidly degraded when the cells were lysed, which together with bioinformatic analyses suggests that these proteins are exposed in the periplasm and anchored to the outer membrane via the PorK lipid. Chemical cross-linking and mass spectrometry analyses confirmed the interaction between PorK and PorN and further revealed that they interact with the PG0189 outer membrane protein. Furthermore, we established that PorN was required for the stable expression of PorK, PorL and PorM. Collectively, these results suggest that the ring-shaped PorK/N complex may form part of the secretion channel of the T9SS. This is the first report showing the structural organization of any T9SS component. PMID:27509186

  10. Activity of oil-formulated conidia of the fungal entomopathogens Nomuraea rileyi and Isaria tenuipes against lepidopterous larvae.

    PubMed

    Vega-Aquino, Paulina; Sanchez-Peña, Sergio; Blanco, Carlos A

    2010-03-01

    The fungi Nomuraea rileyi and Isaria tenuipes (=Paecilomyces tenuipes) are ecologically obligate, widespread pathogens of lepidopterans. Bioassays were carried out to evaluate the activity of oil-suspended conidia of N. rileyi and I. tenuipes against larvae of Spodoptera frugiperda, Spodoptera exigua, Helicoverpa zea, and Heliothis virescens. The tests consisted of two bioassay sets. In the first set, conidia of N. rileyi and I. tenuipes were suspended in water+Tween 80, and in vegetable (canola, soybean) and mineral (proprietary mixture of alkanes and cyclic paraffins) oils, and tested against S. frugiperda. Both fungi were highly compatible with oils and caused mortalities near 100% in all oil treatments; the lowest LT(50) values were 4.7 days for N. rileyi in mineral oil and 6.0 days for I. tenuipes in soybean oil. The second set included additional fungal strains and oil formulations (mineral, canola, sunflower, olive and peanut oils) tested against larvae of S. exigua, S. frugiperda, H. zea and H. virescens. The highest activity was that of N. rileyi in mineral oil against Spodoptera spp., with LT(50) values of 2.5 days (strain ARSEF 135) and 3 days (strain ARSEF 762) respectively. For two different isolates of I. tenuipes the lowest LT(50) values (5.1-5.6 days respectively) were obtained with mineral oil formulations against Spodoptera spp. and H. zea respectively. Additionally, we tested both fungi against prepupae of all four lepidopteran species. Mortalities with I. tenuipes against S. exigua ranged from 90% to 100% (strains ARSEF 2488 and 4096); N. rileyi caused 95% mortality on S. frugiperda. The activity of formulations depended on host species and oil used; Spodoptera spp. was more susceptible to these fungi than Heliothis and Helicoverpa. The results indicate that a comprehensive evaluation of these entomopathogens in agriculture using oil application technologies is advisable, particularly, in organic and sustainable settings. PMID:20025883

  11. Comparison of field-collected ascovirus isolates by DNA hybridization, host range, and histopathology.

    PubMed

    Hamm, J J; Styer, E L; Federici, B A

    1998-09-01

    Six field-collected ascovirus isolates obtained from five noctuid species in the continental United States were compared with respect to the general relatedness of their DNA, host range, and histopathology. Two isolates were from Spodoptera frugiperda, and the other four were from Autographa precationis, Heliothis virescens, Helicoverpa zea, and Trichoplusia ni. DNA-DNA hybridization studies showed that the six isolates belonged to three distinct viral species, with the isolates from S. frugiperda composing one species, those from A. precationis and H. virescens a second species, and those from H. zea and T. ni a third species. The host range and histopathology of each isolate was studied in eight noctuid species, S. frugiperda, Spodoptera ornithogalli, Spodoptera exigua, Spodoptera eridania, H. virescens, H. zea, A. precationis, and Feltia subterranea. Though some variation existed between the different isolates of each viral species, distinct patterns were apparent for each. The viral species from S. frugiperda had a host range that was limited primarily to Spodoptera species and both isolates of this virus only replicated and caused significant pathology in the fat body, whereas the viral species from A. precationis and H. virescens had a much broader host range that included most of the species tested, but also had a tissue tropism primarily restricted to the fat body. The viral species from T. ni and H. zea readily infected all the hosts tested, where the principal site of replication and significant pathology was the epidermis. In many test hosts, however, this viral species also replicated and caused significant pathology in the tracheal epithelium and to a lesser extent in the fat body. Aside from contributing to knowledge of ascovirus biology, these studies indicate that DNA hybridization profiles combined with studies of host range and tissue tropism can be used as characters for defining ascovirus species. PMID:9709014

  12. Nicotiana attenuata MPK4 suppresses a novel JA signaling-independent defense pathway against the specialist insect Manduca sexta but is not required for the resistance to the generalist Spodoptera littoralis

    PubMed Central

    Hettenhausen, Christian; Baldwin, Ian T.; Wu, Jianqiang

    2014-01-01

    Summary How plants tailor their defense responses to attack from different insects remains largely unknown. Here we studied the role of a mitogen-activated protein kinase (MAPK), MPK4, in the resistance of a wild tobacco Nicotiana attenuata to two herbivores, the specialist Manduca sexta and the generalist Spodoptera littoralis. Stably transformed N. attenuata plants silenced in MPK4 (irMPK4) were generated and characterized for traits important for defense against herbivores. Only the oral secretions (OS) from M. sexta, but not the OS from S. littoralis or mechanical wounding, induced elevated levels of jasmonic acid (JA) in irMPK4 plants compared to the wild-type plants. Moreover, silencing MPK4 highly increased the resistance of N. attenuata to M. sexta in a fashion that was independent of COI1 (CORONATINE INSENSITIVE 1)-mediated JA signaling. Untargeted metabolomic screening identified several new MPK4-dependent putative defensive compounds against M. sexta. In contrast, silencing MPK4 did not affect the growth of the generalist insect S. littoralis, and we propose that this was due to the very low levels of fatty acid-amino acid conjugates (FACs) in S. littoralis OS. Thus, MPK4 is likely to be a key signaling element that enables plants to tailor defense responses to different attackers. PMID:23672856

  13. Limonoids from Swietenia macrophylla and S. aubrevilleana.

    PubMed

    Mootoo, B S; Ali, A; Motilal, R; Pingal, R; Ramlal, A; Khan, A; Reynolds, W F; McLean, S

    1999-11-01

    An investigation of the seeds of Swietenia macrophylla and S. aubrevilleana (Meliaceae) is reported. Three new compounds, augustineolide (1) and 3beta,6-dihydroxydihydrocarapin (2) from S. macrophylla and 6-acetoxyhumilinolide C (3) from S. aubrevilleana were isolated and characterized along with fifteen known compounds. Four of the compounds were subjected to an antifeedant bioassay on the final instar larvae of Spodoptera frugiperda. The antifeedant activity was comparable to that of bicyclononanolides previously tested. PMID:10579863

  14. Identification of a delta-Endotoxin Gene Product Specifically Active against Spodoptera littoralis Bdv. among Proteolysed Fractions of the Insecticidal Crystals of Bacillus thuringiensis subsp. aizawai 7.29.

    PubMed

    Lecadet, M M; Sanchis, V; Menou, G; Rabot, P; Lereclus, D; Chaufaux, J; Martouret, D

    1988-11-01

    At least three different insecticidal crystal protein genes were shown to be expressed in Bacillus thuringiensis subsp. aizawai 7.29, a strain that is potentially active against the cotton leafworm Spodoptera littoralis Bdv. Among crude K-60 fractions (60- to 70-kilodalton [kDa] molecules) that were products of proteolysed crystals containing the active domains of the protoxin molecules, we were able to distinguish several distinct components on the basis of their antigenic relationship and their larvicidal properties. A purified fraction designated SF2 was a 61-kDa component specifically active against Pieris brassicae L. and homologous to the B. thuringiensis subsp. berliner 1715 plasmid-encoded crystal protein. A second fraction designated SF1 was composed of 63- and 65-kDa polypeptides and was specifically active against S. littoralis. The SF1 fraction and particularly the 65-kDa component were not antigenically related to the 61-kDa component. The purified fractions were compared with the products of three different crystal protein genes we previously cloned from total DNA of B. thuringiensis subsp. aizawai, among them a new type of crystal protein gene encoding a protein that is specifically active against S. littoralis and other insects of the Noctuidae family. This approach led us to consider the 65-kDa component as a minimum active part of a delta-endotoxin that is encoded by this new gene. Products of the two other cloned genes can be correlated with the 61- and 63-kDa components, respectively. Thus, in B. thuringiensis subsp. aizawai 7.29, multiple delta-endotoxin genes of different structural types direct the synthesis of several delta-endotoxins with different host specificities which were identified as components of the insecticidal crystals. PMID:16347771

  15. The PorX Response Regulator of the Porphyromonas gingivalis PorXY Two-Component System Does Not Directly Regulate the Type IX Secretion Genes but Binds the PorL Subunit

    PubMed Central

    Vincent, Maxence S.; Durand, Eric; Cascales, Eric

    2016-01-01

    The Type IX secretion system (T9SS) is a versatile multi-protein complex restricted to bacteria of the Bacteriodetes phylum and responsible for the secretion or cell surface exposition of diverse proteins that participate to S-layer formation, gliding motility or pathogenesis. The T9SS is poorly characterized but a number of proteins involved in the assembly of the secretion apparatus in the oral pathogen Porphyromonas gingivalis have been identified based on genome substractive analyses. Among these proteins, PorY, and PorX encode typical two-component system (TCS) sensor and CheY-like response regulator respectively. Although the porX and porY genes do not localize at the same genetic locus, it has been proposed that PorXY form a bona fide TCS. Deletion of porX in P. gingivalis causes a slight decrease of the expression of a number of other T9SS genes, including sov, porT, porP, porK, porL, porM, porN, and porY. Here, we show that PorX and the soluble cytoplasmic domain of PorY interact. Using electrophoretic mobility shift, DNA-protein co-purification and heterologous host expression assays, we demonstrate that PorX does not bind T9SS gene promoters and does not directly regulate expression of the T9SS genes. Finally, we show that PorX interacts with the cytoplasmic domain of PorL, a component of the T9SS membrane core complex and propose that the CheY-like PorX protein might be involved in the dynamics of the T9SS.

  16. Expression of recombinant and mosaic Cry1Ac receptors from Helicoverpa armigera and their influences on the cytotoxicity of activated Cry1Ac to Spodoptera litura Sl-HP cells.

    PubMed

    Xu, Peng; Islam, Mayira; Xiao, Yutao; He, Fei; Li, Yi; Peng, Jianxin; Hong, Huazhu; Liu, Chenxi; Liu, Kaiyu

    2016-05-01

    Bacillus thuringiensis (Bt) toxin receptors play important roles in the killing of pests, and investigation on characterization of the receptors is essential for utilization of Bt and management of insect resistance. Here, recombinant and mosaic receptors of Bt Cry1Ac toxin from Helicoverpa armigera were expressed in Spodoptera litura Sl-HP cells and their influences on cytotoxicity of activated Cry1Ac toxin were investigated. When H. armigera aminopeptidase N1 (APN1), alkaline phosphatase 2 (ALP2) and cadherin fused with or without GFP tag were, respectively, expressed in Sl-HP cells, live cell-immunofluorescence staining detection revealed that the quantity of the toxin binding to cadherin or cadherin-GFP was much more than that binding to ALP2 and APN1 or their fusion proteins with GFP, and only the cadherin- or cadherin-GFP-expressing cells showed aberrant cell morphology after the treatment of the toxin at low concentrations. ALP2 and APN1 fused with or without GFP tag did not significantly enhance the cadherin-mediated cytotoxicity of the toxin. The mosaic ALP-TBR-GFP-GPI was located on cell membrane, but did not bind to the toxin. The mosaic truncated cadherin-GFP-GPI was not located on cell membrane even if the signal peptide was sustained. The concentrations of the toxin resulting in swelling of 50 % cells for noncadherin-expressing Sl-HP cells and cadherin-expressing Hi5 cells were 5.08 and 9.50 µg/ml within 1 h, respectively. Taken together, our data have indicated that the binding affinity of ALP2 and APN1 to activated Cry1Ac toxin is much weaker than that of cadherin and both ALP2 and APN1 do not enhance the cytotoxicity of the toxin even though cadherin is co-expressed, and the mosaic receptor of ALP2 inserted with cadherin toxin binding domain does not mediate cytotoxicity of the toxin. In addition, the noncadherin-expressing Sl-HP cells are more susceptible to activated Cry1Ac than the cadherin-expressing Hi5 cells. PMID:25412589

  17. Temperature Impacts the Development and Survival of Common Cutworm (Spodoptera litura): Simulation and Visualization of Potential Population Growth in India under Warmer Temperatures through Life Cycle Modelling and Spatial Mapping

    PubMed Central

    Fand, Babasaheb B.; Sul, Nitin T.; Bal, Santanu K.; Minhas, P. S.

    2015-01-01

    The common cutworm, Spodoptera litura, has become a major pest of soybean (Glycine max) throughout its Indian range. With a changing climate, there is the potential for this insect to become an increasingly severe pest in certain regions due to increased habitat suitability. To examine this possibility, we developed temperature-based phenology model for S. litura, by constructing thermal reaction norms for cohorts of single life stages, at both constant and fluctuating temperatures within the ecologically relevant range (15–38°C) for its development. Life table parameters were estimated stochastically using cohort updating and rate summation approach. The model was implemented in the geographic information system to examine the potential future pest status of S. litura using temperature change projections from SRES A1B climate change scenario for the year 2050. The changes were visualized by means of three spatial indices demonstrating the risks for establishment, number of generations per year and pest abundance according to the temperature conditions. The results revealed that the development rate as a function of temperature increased linearly for all the immature stages of S. litura until approximately 34–36°C, after which it became non-linear. The extreme temperature of 38°C was found lethal to larval and pupal stages of S. litura wherein no development to the next stage occurred. Females could lay no eggs at the extreme low (15°C) and high (> 35°C) test temperatures, demonstrating the importance of optimum temperature in determining the suitability of climate for the mating and reproduction in S. litura. The risk mapping predicts that due to temperature increase under future climate change, much of the soybean areas in Indian states like Madhya Pradesh, Maharashtra and Rajasthan, will become suitable for S. litura establishment and increased pest activity, indicating the expansion of the suitable and favourable areas over time. This has serious

  18. Identification of Porphyromonas gingivalis proteins secreted by the Por secretion system.

    PubMed

    Sato, Keiko; Yukitake, Hideharu; Narita, Yuka; Shoji, Mikio; Naito, Mariko; Nakayama, Koji

    2013-01-01

    The Gram-negative bacterium Porphyromonas gingivalis possesses a number of potential virulence factors for periodontopathogenicity. In particular, cysteine proteinases named gingipains are of interest given their abilities to degrade host proteins and process other virulence factors such as fimbriae. Gingipains are translocated on the cell surface or into the extracellular milieu by the Por secretion system (PorSS), which consists of a number of membrane or periplasmic proteins including PorK, PorL, PorM, PorN, PorO, PorP, PorQ, PorT, PorU, PorV (PG27, LptO), PorW and Sov. To identify proteins other than gingipains secreted by the PorSS, we compared the proteomes of P. gingivalis strains kgp rgpA rgpB (PorSS-proficient strain) and kgp rgpA rgpB porK (PorSS-deficient strain) using two-dimensional gel electrophoresis and peptide-mass fingerprinting. Sixteen spots representing 10 different proteins were present in the particle-free culture supernatant of the PorSS-proficient strain but were absent or faint in that of the PorSS-deficient strain. These identified proteins possessed the C-terminal domains (CTDs), which had been suggested to form the CTD protein family. These results indicate that the PorSS is used for secretion of a number of proteins other than gingipains and that the CTDs of the proteins are associated with the PorSS-dependent secretion. PMID:23075153

  19. System design description for portable 1,000 CFM exhauster Skids POR-007/Skid E and POR-008/Skid F

    SciTech Connect

    Nelson, O.D.

    1998-07-25

    The primary purpose of the two 1,000 CFM Exhauster Skids, POR-007-SKID E and POR-008-SKID F, is to provide backup to the waste tank primary ventilation systems for tanks 241-C-106 and 241-AY-102, and the AY-102 annulus in the event of a failure during the sluicing of tank 241-C-106 and subsequent transfer of sluiced waste to 241-AY-102. This redundancy is required since both of the tank ventilation systems have been declared as Safety Class systems.

  20. Disminuyen en los Estados Unidos las infecciones por VPH.

    Cancer.gov

    La infección por los tipos del virus del papiloma humano (VPH) en el blanco de la vacuna cuadrivalente se redujo en casi dos tercios en las adolescentes desde que se recomendó la vacunación en los Estados Unidos.

  1. Centros oncológicos designados por el NCI

    Cancer.gov

    El programa de centros oncológicos designados por el Instituto Nacional del Cáncer (NCI) reconoce a los centros de todo el país que cumplen con rigurosos criterios para participar en proyectos avanzados de primer nivel para la investigación multidisciplinaria del cáncer.

  2. Se evitaron casi 800 000 muertes por descenso del tabaquismo

    Cancer.gov

    Programas y estrategias de control del tabaco del siglo XX fueron responsables de la prevención de más de 795 000 muertes por cáncer de pulmón en Estados Unidos de 1975 al 2000. Si todo el tabaquismo en este país hubiera cesado después de la publicación d

  3. Functional response of the tiger beetle Megacephala carolina carolina (Coleoptera: Carabidae) on twolined spittlebug (Hemiptera: Cercopidae) and fall armyworm (Lepidoptera: Noctuidae).

    PubMed

    Nachappa, Punya; Braman, S K; Guillebeau, L P; All, J N

    2006-10-01

    The functional response of the tiger beetle Megacephala carolina carolina L. (Coleoptera: Carabidae) was determined on adult twolined spittlebug, Prosapia bicincta (Say) (Hemiptera: Cercopidae), and fourth instars of fall armyworm, Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae), in single-prey and two-prey systems. In the laboratory, M. carolina carolina demonstrated a type II functional response for P. bicincta and S. frugiperda in both single- and two-prey systems. Search efficiency of M. carolina declined for both prey as the initial number of prey increased. Of the total prey consumed, M. carolina carolina killed significantly more S. frugiperda than P. bicincta in the single-prey system (8.0 and 4.5, respectively) and the two-prey system (5.0 and 2.0, respectively). Estimates of attack coefficient, a, were not significantly different for P. bicincta and S. frugiperda in the single-prey (0.07 and 0.02) and two-prey systems (0.04 and 0.06), respectively. The handling time, T(h), was significantly greater for P. bicincta (5.02 and 10.64 h) than for S. frugiperda (2.66 and 4.41 h) in single- and two-prey systems, respectively. Estimations of attack coefficient and handling time in the single-prey system were used to predict prey preference of M. carolina carolina. No strong prey switching response was observed. M. carolina carolina showed no preference for either prey. However, in the presence of S. frugiperda, the functional response of the predator for P. bicincta was reduced. M. carolina carolina is a potential predator of one or more turfgrass pests and should be considered in conservation efforts. PMID:17066786

  4. 3-β-D-Glucopyranosyl-6-methoxy-2-benzoxazolinone (MBOA-N-Glc) is an insect detoxification product of maize 1,4-benzoxazin-3-ones.

    PubMed

    Maag, Daniel; Dalvit, Claudio; Thevenet, Damien; Köhler, Angela; Wouters, Felipe C; Vassão, Daniel G; Gershenzon, Jonathan; Wolfender, Jean-Luc; Turlings, Ted C J; Erb, Matthias; Glauser, Gaetan

    2014-06-01

    In order to defend themselves against arthropod herbivores, maize plants produce 1,4-benzoxazin-3-ones (BXs), which are stored as weakly active glucosides in the vacuole. Upon tissue disruption, BXs come into contact with β-glucosidases, resulting in the release of active aglycones and their breakdown products. While some aglycones can be reglucosylated by specialist herbivores, little is known about how they detoxify BX breakdown products. Here we report on the structure of an N-glucoside, 3-β-d-glucopyranosyl-6-methoxy-2-benzoxazolinone (MBOA-N-Glc), purified from Spodoptera frugiperda faeces. In vitro assays showed that MBOA-N-Glc is formed enzymatically in the insect gut using the BX breakdown product 6-methoxy-2-benzoxazolinone (MBOA) as precursor. While Spodoptera littoralis and S. frugiperda caterpillars readily glucosylated MBOA, larvae of the European corn borer Ostrinia nubilalis were hardly able to process the molecule. Accordingly, Spodoptera caterpillar growth was unaffected by the presence of MBOA, while O. nubilalis growth was reduced. We conclude that glucosylation of MBOA is an important detoxification mechanism that helps insects tolerate maize BXs. PMID:24713572

  5. Neisseria meningitidis Lacking the Major Porins PorA and PorB Is Viable and Modulates Apoptosis and the Oxidative Burst of Neutrophils.

    PubMed

    Peak, Ian R; Chen, Adrienne; Jen, Freda E-C; Jennings, Courtney; Schulz, Benjamin L; Saunders, Nigel J; Khan, Arshad; Seifert, H Steven; Jennings, Michael P

    2016-08-01

    The bacterial pathogen Neisseria meningitidis expresses two major outer-membrane porins. PorA expression is subject to phase-variation (high frequency, random, on-off switching), and both PorA and PorB are antigenically variable between strains. PorA expression is variable and not correlated with meningococcal colonisation or invasive disease, whereas all naturally-occurring strains express PorB suggesting strong selection for expression. We have generated N. meningitidis strains lacking expression of both major porins, demonstrating that they are dispensable for bacterial growth in vitro. The porAB mutant strain has an exponential growth rate similar to the parental strain, as do the single porA or porB mutants, but the porAB mutant strain does not reach the same cell density in stationary phase. Proteomic analysis suggests that the double mutant strain exhibits compensatory expression changes in proteins associated with cellular redox state, energy/nutrient metabolism, and membrane stability. On solid media, there is obvious growth impairment that is rescued by addition of blood or serum from mammalian species, particularly heme. These porin mutants are not impaired in their capacity to inhibit both staurosporine-induced apoptosis and a phorbol 12-myristate 13-acetate-induced oxidative burst in human neutrophils suggesting that the porins are not the only bacterial factors that can modulate these processes in host cells. PMID:26562068

  6. Compton imaging with the PorGamRays spectrometer

    NASA Astrophysics Data System (ADS)

    Judson, D. S.; Boston, A. J.; Coleman-Smith, P. J.; Cullen, D. M.; Hardie, A.; Harkness, L. J.; Jones, L. L.; Jones, M.; Lazarus, I.; Nolan, P. J.; Pucknell, V.; Rigby, S. V.; Seller, P.; Scraggs, D. P.; Simpson, J.; Slee, M.; Sweeney, A.; PorGamRays Collaboration

    2011-10-01

    The PorGamRays project aims to develop a portable gamma-ray detection system with both spectroscopic and imaging capabilities. The system is designed around a stack of thin Cadmium Zinc Telluride (CZT) detectors. The imaging capability utilises the Compton camera principle. Each detector is segmented into 100 pixels which are read out through custom designed Application Specific Integrated Circuits (ASICs). This device has potential applications in the security, decommissioning and medical fields. This work focuses on the near-field imaging performance of a lab-based demonstrator consisting of two pixelated CZT detectors, each of which is bonded to a NUCAM II ASIC. Measurements have been made with point 133Ba and 57Co sources located ˜35 mm from the surface of the scattering detector. Position resolution of ˜20 mm FWHM in the x and y planes is demonstrated.

  7. Acceptance test report for portable exhauster POR-007/Skid E

    SciTech Connect

    Kriskovich, J.R.

    1998-07-24

    This document describes Acceptance Testing performed on Portable Exhauster POR-007/Skid E. It includes measurements of bearing vibration levels, pressure decay testing, programmable logic controller interlocks, high vacuum, flow and pressure control functional testing. The purpose of Acceptance testing documented by this report was to demonstrate compliance of the exhausters with the performance criteria established within HNF-0490, Rev. 1 following a repair and upgrade effort at Hanford. In addition, data obtained during this testing is required for the resolution of outstanding Non-conformance Reports (NCR), and finally, to demonstrate the functionality of the associated software for the pressure control and high vacuum exhauster operating modes provided for by W-320. Additional testing not required by the ATP was also performed to assist in the disposition and close out of receiving inspection report and for application design information (system curve). Results of this testing are also captured within this document.

  8. Typing and surface charges of the variable loop regions of PorB from Neisseria meningitidis.

    PubMed

    Stefanelli, Paola; Neri, Arianna; Tanabe, Mikio; Fazio, Cecilia; Massari, Paola

    2016-06-01

    PorB is a pan-Neisserial major outer membrane protein with a trimeric β-barrel structure. Each monomer presents eight periplasmic turns and eight surface exposed loop regions with sequence variability. PorB induces activation of host cell responses via a TLR2-dependent mechanism likely mediated by electrostatic interactions between TLR2 and PorB surface exposed loops. Variability in the loop amino acid sequence is known to influence cell responses to PorB in vitro, particularly for the residues in L5 and L7. In this work, the sequence of the porB gene and the electrostatic surface charges of PorB from 35 invasive meningococcal isolates belonging to the main clonal complexes identified in Italy and from five carriage genomes available on the website http://pubmlst.org/neisseria/ were examined. Analysis of the porB encoding regions from the invasive meningococci has identified four new alleles and a potential association between porB alleles, serogroup, and clonal complexes. Through computer-based modeling and analysis of the electrostatic surface charges of PorB from these strains, loop charge segregation between PorB from invasive serogroups B and C was observed. Specifically, loops 1, 4, and 7 were negatively charged and L2 and L8 were mostly neutral in serogroup B isolates, while an overall homogeneous positive surface charge was present in PorB from invasive serogroup C strains. A higher PorB sequence variability was observed among carriage genomes, and a general prevalence of negative loop surface charges. The surface charge differences in PorB from serogroups B and C invasive and carriage strains may, in part, influence the outcomes of Neisseriae interactions with host cells. © 2016 IUBMB Life, 68(6):488-495, 2016. PMID:27156582

  9. [Pathogenicity and genetic divergence of two isolates of microsporidia Nosema bombycis].

    PubMed

    WANG, Min; XU, Jin-Shan; WANG, Lin-Ling; ZHANG, Xiao-Yan; ZHOU, Ze-Yang

    2009-11-01

    Microsporidia Nosema bombycis is a fungal pathogen that causes epidemic pebrine disease in Bombyx mori. Two N. bombycis isolates were obtained from two areas in China and showed different pathogenicity after Spodoptera frugiperda Sf9 cell cultivation. The regions of rDNAs from different isolates were analyzed, suggesting no relationship between the genetic divergence and their geographic distributions. Further analysis showed that several copies of SSU rDNA units in N. bombycis were interrupted by a MITE-like transposon, indicating the complexity of genomic structure in Nosema bombycis. PMID:19933093

  10. Effects of limonoids from Cipadessa fruticosa on fall armyworm.

    PubMed

    Matos, Andreia P; Leite, Ana C; Batista-Pereira, Luciane G; Vieira, Paulo C; Fernandes, Josäo B; da Silva, Maria Fátima das G F

    2009-01-01

    Six mexicanolide limonoids isolated from the dichloromethane extract of the fruits of Cipadessa fruticosa Blume (Meliaceae) were evaluated against Spodoptera frugiperda (J. E. Smith). Gedunin was used as a positive control. When incorporated into an artificial diet of neonates at 50.0 mg kg(-1), febrifugin A showed 73.3% mortality. All the compounds showed moderate insecticidal activity, except for ruageanin A, when compared with the control. Febrifugin also showed growth inhibition and antifeedant activities (at 100.0 mg kg(-1)). The correlation between the insecticidal activity of the isolated compounds and their chemical structure was discussed. PMID:19678552

  11. Acceptance test report for portable exhauster POR-008/Skid F

    SciTech Connect

    Kriskovich, J.R.

    1998-07-24

    Portable Exhauster POR-008 was procured via HNF-0490, Specification for a Portable Exhausted System for Waste Tank Ventilation. Prior to taking ownership, acceptance testing was performed at the vendors. However at the conclusion of testing a number of issues remained that required resolution before the exhausters could be used by Project W-320. The purpose of acceptance testing documented by this report was to demonstrate compliance of the exhausters with the performance criteria established within HNF-O49O, Rev. 1 following a repair and upgrade effort at Hanford. In addition, data obtained during this testing is required for the resolution of outstanding Non-conformance Reports (NCR), and finally, to demonstrate the functionality of the associated software for the pressure control and high vacuum exhauster operating modes provided for by W-320. Additional testing not required by the ATP was also performed to assist in the disposition and close out of receiving inspection report and for application design information (system curve). Results of this testing are also captured within this document.

  12. VDAC and the bacterial porin PorB of Neisseria gonorrhoeae share mitochondrial import pathways.

    PubMed

    Müller, Anne; Rassow, Joachim; Grimm, Jan; Machuy, Nikolaus; Meyer, Thomas F; Rudel, Thomas

    2002-04-15

    The human pathogen Neisseria gonorrhoeae induces host cell apoptosis during infection by delivering the outer membrane protein PorB to the host cell's mitochondria. PorB is a pore-forming beta-barrel protein sharing several features with the mitochondrial voltage-dependent anion channel (VDAC), which is involved in the regulation of apoptosis. Here we show that PorB of pathogenic Neisseria species produced by host cells is efficiently targeted to mitochondria. Imported PorB resides in the mitochondrial outer membrane and forms multimers with similar sizes as in the outer bacterial membrane. The mitochondria completely lose their membrane potential, a characteristic previously observed in cells infected with gonococci or treated with purified PorB. Closely related bacterial porins of non-pathogenic Neisseria mucosa or Escherichia coli remain in the cytosol. Import of PorB into mitochondria in vivo is independent of a linear signal sequence. Insertion of PorB into the mitochondrial outer membrane in vitro depends on the activity of Tom5, Tom20 and Tom40, but is independent of Tom70. Our data show that human VDAC and bacterial PorB are imported into mitochondria by a similar mechanism. PMID:11953311

  13. Comparative analysis of expression profiling of the trypsin and chymotrypsin genes from Lepidoptera species with different levels of sensitivity to soybean peptidase inhibitors.

    PubMed

    Souza, Thais P; Dias, Renata O; Castelhano, Elaine C; Brandão, Marcelo M; Moura, Daniel S; Silva-Filho, Marcio C

    2016-01-01

    Peptidase inhibitors (PIs) are essential proteins involved in plant resistance to herbivorous insects, yet many insect species are able to escape the negative effects of these molecules. We compared the effects of acute and chronic ingestion of soybean peptidase inhibitors (SPIs) on Spodoptera frugiperda and Diatraea saccharalis, two Lepidoptera species with different sensitivities to SPI ingestion. We analyzed the trypsin and chymotrypsin gene expression profiles in both species. Acute exposure of S. frugiperda to the inhibitors activated seven genes (SfChy5, SfChy9, SfChy19, SfChy22, SfTry6, SfTry8, and SfTry10), whereas chronic exposure activated 16 genes (SfChy2, SfChy4, SfChy5, SfChy8, SfChy9, SfChy11, SfChy12, SfChy15, SfChy17, SfChy21, SfChy22, SfTry6, SfTry8, SfTry9, SfTry10, and SfTry12). By contrast, the challenge of D. saccharalis with SPIs did not differentially induce the expression of trypsin- or chymotrypsin-encoding genes, with the exception of DsChy7. Bayesian phylogenetic analysis of S. frugiperda trypsin protein sequences revealed two gene clades: one composed of genes responsive to the SPIs and a second composed of the unresponsive genes. D. saccharalis trypsin proteins were clustered nearest to the S. frugiperda unresponsive genes. Overall, our findings support a hypothesized mechanism of resistance of Noctuidae moths to SPIs, involving gene number expansion of trypsin and chymotrypsin families and regulation of gene expression, which could also explain the variable susceptibility between S. frugiperda and D. saccharalis to these plant inhibitors. PMID:26944308

  14. Intraguild Competition of Three Noctuid Maize Pests.

    PubMed

    Bentivenha, J P F; Baldin, E L L; Hunt, T E; Paula-Moraes, S V; Blankenship, E E

    2016-08-01

    The western bean cutworm Striacosta albicosta (Smith), the fall armyworm Spodoptera frugiperda (J. E. Smith), and the corn earworm Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae) are among the major lepidopteran pests of maize in the United States, belonging to the same guild and injuring the reproductive tissues of this crop. Here, intraguild competition of these lepidopterans on non-Bt maize was evaluated through survival analysis of each species under laboratory and field conditions. Competition scenarios were carried out in arenas containing maize silk or ear tissue, using larvae on different stadium of development. Fitness cost competition studies were conducted to examine the influence of intraguild competition and cannibalism and predation rates on larval development. The survival of S. albicosta competing with the other species was significantly lower than in intraspecific competition, even when the larvae were more developed than the competitor. For S. frugiperda, survival remained high in the different competition scenarios, except when competing in a smaller stadium with H. zea Larvae of H. zea had a high rate of cannibalism, higher survival when competing against S. albicosta than S. frugiperda, and reduced survival when the H. zea larvae were at the same development stadium or smaller than the competitors. Based on fitness cost results, the absence of a competitor for the feeding source may confer an advantage to the larval development of S. frugiperda and H. zea Our data suggest that S. frugiperda has a competitive advantage against the other species, while S. albicosta has the disadvantage in the intraguild competition on non-Bt maize. PMID:27330146

  15. Effects of bacillus thuringiensis transgenic corn on corn earworm and fall armyworm (Lepidoptera: Noctuidae) densities.

    PubMed

    Chilcutt, Charles F; Odvody, Gary N; Correa, J Carlos; Remmers, Jeff

    2007-04-01

    We examined 17 pairs of near-isogenic hybrids of Bacillus thuringiensis (Bt) (176, Mon810, and Bt11) and non-Bt corn, Zea mays L., to examine the effects of Bt on larval densities of Helicoverpa zea (Boddie) and Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae) during 2 yr. During ear formation, instar densities of H. zea and S. frugiperda were recorded for each hybrid. We found that H. zea first, second, and fifth instar densities were each affected by Mon810 and Bt11 Bt corn but not by 176 corn. Surprisingly, first and second instars were found in higher numbers on ears of Mon810 and Bt11 corn than on non-Bt corn. Densities of third and fourth instars were equal on Bt and non-Bt hybrids, whereas densities of fifth instars were lower on Bt plants. S. frugiperda larval densities were only affected during 1 yr when second, and fourth to sixth instars were lower on ears of Mon810 and Bt11 hybrids compared with their non-Bt counterparts. Two likely explanations for early instar H. zea densities being higher on Bt corn than non-Bt corn are that (1) Bt toxins delay development, creating a greater abundance of early instars that eventually die, and (2) reduced survival of H. zea to later instars on Bt corn decreased the normal asymmetric cannibalism or H. zea-S. frugiperda intraguild predation of late instars on early instars. Either explanation could explain why differences between Bt and non-Bt plants were greater for H. zea than S. frugiperda, because H. zea is more strongly affected by Bt toxins and more cannibalistic. PMID:17461054

  16. Successful Mnemonics for "por"/"para" and Affirmative Commands with Pronouns.

    ERIC Educational Resources Information Center

    Mason, Keith

    1992-01-01

    Two mnemonic devices, "4A Rule" and "PERFECT," are described to simplify the learning of two grammar points: the placement of object pronouns with respect to commands and the distinction between "por" and "para." (five references) (LB)

  17. Differential requirements of two insect cell lines for growth in serum-free medium.

    PubMed

    Vaughn, J L; Fan, F

    1997-06-01

    The development of a serum-free medium that supports the growth of cells from a Spodoptera frugiperda and a Lymantria dispar cell line is reported. A yeast hydrolysate provided the B-vitamin complex, and a combination of a meat hydrolysate and tryptose provided most of the free amino acids required for cell growth. Supplemental cystine and methionine were required to achieve maximum cell growth. The serum or serum replacements used in earlier formulations were replaced with commercial lipid preparations and increased levels of iron salts. Although the cell growth cycle had a somewhat extended lag phase and the population doubling time of the S. frugiperda cells was longer than on serum-containing medium, the saturation densities were much higher. Spodoptera cells grown in this medium replicated the Autographa californica nuclear polyhedrosis virus well, producing 8.71 x 10(6) TCID50 extracellular virus and 4.4 x 10(6) polyhedra/ml culture. The specific activity of the polyhedra was somewhat less than that of polyhedra produced in insects. PMID:9201517

  18. Field and laboratory evaluations of transgenic cottons expressing one or two Bacillus thuringiensis var. kurstaki Berliner proteins for management of noctuid (Lepidoptera) pests.

    PubMed

    Chitkowski, R L; Turnipseed, S G; Sullivan, M J; Bridges, W C

    2003-06-01

    Field studies were conducted from 1999 to 2001 to evaluate the efficacy of the transgenic cotton, Gossypium hirsutum (L.), genotype, Bollgard II (Monsanto 15985), which expresses two Bacillus thuringiensis Berliner (Bt) proteins (Cry1Ac + Cry2Ab) that are active against lepidopterous pests. Bollgard II was compared with Bollgard (DP50B), which expresses only one Bt protein (Cry1Ac), and, in all tests, the conventional variety, DP50, was used as a non-Bt control. Larval populations of the bollworm, Helicoverpa zea (Boddie), and the soybean looper, Pseudoplusia includens (Walker), were significantly lower in Bollgard II than in Bollgard and conventional cotton, and the proportion of fruit damaged by H. zea was also lower. Fall armyworm, Spodoptera frugiperda (J. E. Smith), populations were lower in Bollgard II than in Bollgard, although not significantly. Field tests were supplemented with laboratory bioassays in 2001 to compare mortality of S. frugiperda, and beet armyworms, Spodoptera exigua (Hübner), feeding on these genotypes. Mortality of both species was significantly greater on Bollgard II plant material than on either Bollgard or conventional cotton. This study demonstrated that the dual-toxin Bollgard II genotype is highly effective against lepidopterous pests that are not adequately controlled by the current single-toxin Bollgard varieties. If toxin expression in future Bollgard II varieties remains consistent with that of Monsanto 15985, supplemental insecticides will be reduced, and may be eliminated for lepidopterous pests in South Carolina. PMID:12852613

  19. Genome Sequences of Pseudomonas oryzihabitans Phage POR1 and Pseudomonas aeruginosa Phage PAE1.

    PubMed

    Dyson, Zoe A; Seviour, Robert J; Tucci, Joseph; Petrovski, Steve

    2016-01-01

    We report the genome sequences of two double-stranded DNA siphoviruses, POR1 infective for Pseudomonas oryzihabitans and PAE1 infective for Pseudomonas aeruginosa The phage POR1 genome showed no nucleotide sequence homology to any other DNA phage sequence in the GenBank database, while phage PAE1 displayed synteny to P. aeruginosa phages M6, MP1412, and YuA. PMID:27313312

  20. Genome Sequences of Pseudomonas oryzihabitans Phage POR1 and Pseudomonas aeruginosa Phage PAE1

    PubMed Central

    Dyson, Zoe A.; Seviour, Robert J.; Tucci, Joseph

    2016-01-01

    We report the genome sequences of two double-stranded DNA siphoviruses, POR1 infective for Pseudomonas oryzihabitans and PAE1 infective for Pseudomonas aeruginosa. The phage POR1 genome showed no nucleotide sequence homology to any other DNA phage sequence in the GenBank database, while phage PAE1 displayed synteny to P. aeruginosa phages M6, MP1412, and YuA. PMID:27313312

  1. Structural basis for solute transport, nucleotide regulation, and immunological recognition of Neisseria meningitidis PorB

    PubMed Central

    Tanabe, Mikio; Nimigean, Crina M.; Iverson, T. M.

    2010-01-01

    PorB is the second most prevalent outer membrane protein in Neisseria meningitidis. PorB is required for neisserial pathogenesis and can elicit a Toll-like receptor mediated host immune response. Here, the x-ray crystal structure of PorB has been determined to 2.3 Å resolution. Structural analysis and cocrystallization studies identify three putative solute translocation pathways through the channel pore: One pathway transports anions nonselectively, one transports cations nonselectively, and one facilitates the specific uptake of sugars. During infection, PorB likely binds host mitochondrial ATP, and cocrystallization with the ATP analog AMP–PNP suggests that binding of nucleotides regulates these translocation pathways both by partial occlusion of the pore and by restricting the motion of a putative voltage gating loop. PorB is located on the surface of N. meningitidis and can be recognized by receptors of the host innate immune system. Features of PorB suggest that Toll-like receptor mediated recognition outer membrane proteins may be initiated with a nonspecific electrostatic attraction. PMID:20351243

  2. An investigation of exploitation versus exploration in GBEA optimization of PORS 15 and 16 Problems

    SciTech Connect

    Koch, Kaelynn

    2012-01-01

    It was hypothesized that the variations in time to solution are driven by the competing mechanisms of exploration and exploitation.This thesis explores this hypothesis by examining two contrasting problems that embody the hypothesized tradeoff between exploration and exploitation. Plus one recall store (PORS) is an optimization problem based on the idea of a simple calculator with four buttons: plus, one, store, and recall. Integer addition and store are classified as operations, and one and memory recall are classified as terminals. The goal is to arrange a fixed number of keystrokes in a way that maximizes the numerical result. PORS 15 (15 keystrokes) represents the subset of difficult PORS problems and PORS 16 (16 keystrokes) represents the subset of PORS problems that are easiest to optimize. The goal of this work is to examine the tradeoff between exploitation and exploration in graph based evolutionary algorithm (GBEA) optimization. To do this, computational experiments are used to examine how solutions evolve in PORS 15 and 16 problems when solved using GBEAs. The experiment is comprised of three components; the graphs and the population, the evolutionary algorithm rule set, and the example problems. The complete, hypercube, and cycle graphs were used for this experiment. A fixed population size was used.

  3. Crystallographic analysis of Neisseria meningitidis PorB extracellular loops potentially implicated in TLR2 recognition.

    PubMed

    Kattner, Christof; Toussi, Deana N; Zaucha, Jan; Wetzler, Lee M; Rüppel, Nadine; Zachariae, Ulrich; Massari, Paola; Tanabe, Mikio

    2014-03-01

    Among all Neisseriae species, Neisseria meningitidis and Neisseria gonorrhoeae are the only human pathogens, causative agents of bacterial meningitis and gonorrhoea, respectively. PorB, a pan-Neisseriae trimeric porin that mediates diffusive transport of essential molecules across the bacterial outer membrane, is also known to activate host innate immunity via Toll-like receptor 2 (TLR2)-mediated signaling. The molecular mechanism of PorB binding to TLR2 is not known, but it has been hypothesized that electrostatic interactions contribute to ligand/receptor binding. Strain-specific sequence variability in the surface-exposed loops of PorB which are potentially implicated in TLR2 binding, may explain the difference in TLR2-mediated cell activation in vitro by PorB homologs from the commensal Neisseriae lactamica and the pathogen N. meningitidis. Here, we report a comparative structural analysis of PorB from N. meningitidis serogroup B strain 8765 (63% sequence homology with PorB from N. meningitidis serogroup W135) and a mutant in which amino acid substitutions in the extracellular loop 7 lead to significantly reduced TLR2-dependent activity in vitro. We observe that this mutation both alters the loop conformation and causes dramatic changes of electrostatic surface charge, both of which may affect TLR2 recognition and signaling. PMID:24361688

  4. Structural basis for solute transport, nucleotide regulation, and immunological recognition of Neisseria meningitidis PorB

    SciTech Connect

    Tanabe, Mikio; Nimigean, Crina M.; Iverson, T.M.

    2010-06-25

    PorB is the second most prevalent outer membrane protein in Neisseria meningitidis. PorB is required for neisserial pathogenesis and can elicit a Toll-like receptor mediated host immune response. Here, the x-ray crystal structure of PorB has been determined to 2.3 {angstrom} resolution. Structural analysis and cocrystallization studies identify three putative solute translocation pathways through the channel pore: One pathway transports anions nonselectively, one transports cations nonselectively, and one facilitates the specific uptake of sugars. During infection, PorB likely binds host mitochondrial ATP, and cocrystallization with the ATP analog AMP-PNP suggests that binding of nucleotides regulates these translocation pathways both by partial occlusion of the pore and by restricting the motion of a putative voltage gating loop. PorB is located on the surface of N. meningitidis and can be recognized by receptors of the host innate immune system. Features of PorB suggest that Toll-like receptor mediated recognition outer membrane proteins may be initiated with a nonspecific electrostatic attraction.

  5. Insecticidal Properties of a Highly Potent Wax Isolated from Dolichandra cynanchoides Cham.

    PubMed

    Díaz Napal, Georgina; Carpinella, María C; Palacios, Sara M

    2016-01-01

    Bioassay-guided fractionation of an ethanolic extract of the aerial parts of Dolichandra cynanchoides Cham. (Bignoniaceae) led to the isolation of a natural wax with anti-insect activity against Spodoptera frugiperda (Noctuidae) and Epilachna paenulata (Coleptera). The compound was identified spectroscopically as an ester of a C27 fatty acid and a C25 alcohol, pentacosyl heptacosanoate (1). The effective doses of 1 for 50% feeding inhibition (ED50) of S. frugiperda and E. paenulata were 0.82 and 8.53 µg/cm², respectively, in a choice test, while azadirachtin showed ED50 of 0.10 and 0.59 µg/cm², respectively. In a no-choice test, both insects refused to feed on leaves treated with 1 at doses of 0.1 µg/cm² or greater inhibiting larval growth and dramatically reducing survival. The lethal doses 50 (LD50) of 1 were 0.39 and 0.68 µg/cm² for S. frugiperda and E. paenulata, respectively. These results indicate that 1 has potential for development as botanical insecticides. Similar esters might be obtainable in large quantities as many edible crops produce wax esters that are discarded during food processing. Research on these materials could lead to the detection of similar waxes with insecticidal activity. PMID:27529199

  6. Fragments of ATP synthase mediate plant perception of insect attack

    PubMed Central

    Schmelz, Eric A.; Carroll, Mark J.; LeClere, Sherry; Phipps, Stephen M.; Meredith, Julia; Chourey, Prem S.; Alborn, Hans T.; Teal, Peter E. A.

    2006-01-01

    Plants can perceive a wide range of biotic attackers and respond with targeted induced defenses. Specificity in plant non-self-recognition occurs either directly by perception of pest-derived elicitors or indirectly through resistance protein recognition of host targets that are inappropriately proteolyzed. Indirect plant perception can occur during interactions with pathogens, yet evidence for analogous events mediating the detection of insect herbivores remains elusive. Here we report indirect perception of herbivory in cowpea (Vigna unguiculata) plants attacked by fall armyworm (Spodoptera frugiperda) larvae. We isolated and identified a disulfide-bridged peptide (+ICDINGVCVDA−), termed inceptin, from S. frugiperda larval oral secretions that promotes cowpea ethylene production at 1 fmol leaf−1 and triggers increases in the defense-related phytohormones salicylic acid and jasmonic acid. Inceptins are proteolytic fragments of chloroplastic ATP synthase γ-subunit regulatory regions that mediate plant perception of herbivory through the induction of volatile, phenylpropanoid, and protease inhibitor defenses. Only S. frugiperda larvae that previously ingested chloroplastic ATP synthase γ-subunit proteins and produced inceptins significantly induced cowpea defenses after herbivory. Digestive fragments of an ancient and essential plant enzyme, inceptin functions as a potent indirect signal initiating specific plant responses to insect attack. PMID:16720701

  7. Cry1F resistance among lepidopteran pests: a model for improved resistance management?

    PubMed

    Vélez, Ana M; Vellichirammal, Neetha Nanoth; Jurat-Fuentes, Juan Luis; Siegfried, Blair D

    2016-06-01

    The Cry1Fa protein from the bacterium Bacillus thuringiensis (Bt) is known for its potential to control lepidopteran pests, especially through transgenic expression in maize and cotton. The maize event TC1507 expressing the cry1Fa toxin gene became commercially available in the United States in 2003 for the management of key lepidopteran pests including the European corn borer, Ostrinia nubilalis, and the fall armyworm, Spodoptera frugiperda. A high-dose/refuge strategy has been widely adopted to delay evolution of resistance to event TC1507 and other transgenic Bt crops. Efficacy of this strategy depends on the crops expressing a high dose of the Bt toxin to targeted pests and adjacent refuges of non-Bt host plants serving as a source of abundant susceptible insects. While this strategy has proved effective in delaying O. nubilalis resistance, field-evolved resistance to event TC1507 has been reported in S. frugiperda populations in Puerto Rico, Brazil, and the southeastern United States. This paper examines available information on resistance to Cry1Fa in O. nubilalis and S. frugiperda and discusses how this information identifies opportunities to refine resistance management recommendations for Bt maize. PMID:27436741

  8. Population dynamics of caterpillars on three cover crops before sowing cotton in Mato Grosso (Brazil).

    PubMed

    Silvie, P J; Menzel, C A; Mello, A; Coelho, A G

    2010-01-01

    Direct seeding mulch-based cropping systems under a preliminary cover crop such as millet are common in some areas of Brazil. Lepidopteran pests that damage cotton, soybean and maize crops can proliferate on cover crops, so preventive chemical treatments are necessary. Very little data is available on these pests on cover crops. This paper presents the dynamics of Spodoptera frugiperda, S. eridania, Mocis latipes and Diatraea saccharalis caterpillars monitored at Primavera do Leste, Mato Grosso state (Brazil) during the of 2005/2006 and 2006/2007 cropping seasons on four cover crops, i.e. finger millet (Eleusine coracana), pearl millet (Pennisetum glaucum), sorghum (Sorghum bicolor) and ruzigrass (Brachiaria ruziziensis). The pests were visually counted on plants within a 1 m2 transect (wooden frame). Caterpillars were reared to facilitate identification of collected species and parasitoids. Many S. frugiperda caterpillars were observed on millet in 2005, with a maximum of 37 caterpillars/m2. On sorghum, we found 30 caterpillars/m2, or 0.83 caterpillars/plant. The Diatraea borer attacked sorghum later than the other pests. M. latipes was also observed on millet. The millet cover crop had to be dried for at least 1 month before direct drilling the main cotton crop in order to impede S. frugiperda infestations on cotton plantlets, thus avoiding the need for substantial resowing. The comparative methodological aspects are discussed. PMID:21539250

  9. Characterization of Nuclear Polyhedrosis Virus DNAs

    PubMed Central

    Summers, Max D.; Anderson, David L.

    1973-01-01

    The nuclear polyhedrosis virus DNAs characterized and compared in this study consist of the singly-enveloped nucleocapsids (SNPV) of Trichoplusia ni and the bundles of nucleocapsids common to a single envelope (MNPV) from Spodoptera frugiperda and Rachiplusia ou. The SNPV and MNPV DNAs are very similar in hydrodynamic properties and molecular weights. In addition, the NPV DNAs are similar in size to those extracted from the granulosis viruses that infect T. ni and S. frugiperda. As isolated from purified virus or directly from occluded virus, the nuclear polyhedrosis virus DNAs consist of a mixture of about 20 to 30% double-stranded covalently closed molecules and approximately 60% relaxed circles, with less than 10% in linear duplex form. The molecular weights of all nuclear polyhedrosis virus DNAs as compared in this study are slightly smaller than those of T4 bacteriophage DNA and perhaps slightly smaller than those of the granulosis virus DNAs. The best estimates of these molecular weights by neutral sucrose sedimentation for the nuclear polyhedrosis viruses range from 90 to 100 × 106 relative to a size of 108 × 106 for T4 DNA. The base compositions of the nuclear polyhedrosis viruses that infect T. ni and S. frugiperda are compared with the respective insect host DNAs. Images PMID:4761726

  10. Molecular characterisation of Porcine rubulavirus (PorPV) isolates from different outbreaks in Mexico.

    PubMed

    Cuevas-Romero, S; Rivera-Benítez, J F; Blomström, A-L; Ramliden, M; Hernández-Baumgarten, E; Hernández-Jáuregui, P; Ramírez-Mendoza, H; Berg, M

    2016-02-01

    Since the report of the initial outbreak of Porcine rubulavirus (PorPV) infection in pigs, only one full-length genome from 1984 (PorPV-LPMV/1984) has been characterised. To investigate the overall genetic variation, full-length gene nucleotide sequences of current PorPV isolates were obtained from different clinical cases of infected swine. Genome organisation and sequence analysis of the encoded proteins (NP, P, F, M, HN and L) revealed high sequence conservation of the NP protein and the expression of the P and V proteins in all PorPV isolates. The V protein of one isolate displayed a mutation that has been implicated to antagonise the antiviral immune responses of the host. The M protein indicated a variation in a short region that could affect the electrostatic charge and the interaction with the membrane. One PorPV isolate recovered from the lungs showed a mutation at the cleavage site (HRKKR) of the F protein that could represent an important factor to determine the tissue tropism and pathogenicity of this virus. The HN protein showed high sequence identity through the years (up to 2013). Additionally, a number of sequence motifs of very high amino acid conservation among the PorPV isolates important for polymerase activity of the L protein have been identified. In summary, genetic comparisons and phylogenetic analyses indicated that three different genetic variants of PorPV are currently spreading within the swine population, and a new generation of circulating virus with different characteristics has begun to emerge. PMID:26728078

  11. Por Secretion System-Dependent Secretion and Glycosylation of Porphyromonas gingivalis Hemin-Binding Protein 35

    PubMed Central

    Shoji, Mikio; Sato, Keiko; Yukitake, Hideharu; Kondo, Yoshio; Narita, Yuka; Kadowaki, Tomoko; Naito, Mariko; Nakayama, Koji

    2011-01-01

    The anaerobic Gram-negative bacterium Porphyromonas gingivalis is a major pathogen in severe forms of periodontal disease and refractory periapical perodontitis. We have recently found that P. gingivalis has a novel secretion system named the Por secretion system (PorSS), which is responsible for secretion of major extracellular proteinases, Arg-gingipains (Rgps) and Lys-gingipain. These proteinases contain conserved C-terminal domains (CTDs) in their C-termini. Hemin-binding protein 35 (HBP35), which is one of the outer membrane proteins of P. gingivalis and contributes to its haem utilization, also contains a CTD, suggesting that HBP35 is translocated to the cell surface via the PorSS. In this study, immunoblot analysis of P. gingivalis mutants deficient in the PorSS or in the biosynthesis of anionic polysaccharide-lipopolysaccharide (A-LPS) revealed that HBP35 is translocated to the cell surface via the PorSS and is glycosylated with A-LPS. From deletion analysis with a GFP-CTD[HBP35] green fluorescent protein fusion, the C-terminal 22 amino acid residues of CTD[HBP35] were found to be required for cell surface translocation and glycosylation. The GFP-CTD fusion study also revealed that the CTDs of CPG70, peptidylarginine deiminase, P27 and RgpB play roles in PorSS-dependent translocation and glycosylation. However, CTD-region peptides were not found in samples of glycosylated HBP35 protein by peptide map fingerprinting analysis, and antibodies against CTD-regions peptides did not react with glycosylated HBP35 protein. These results suggest both that the CTD region functions as a recognition signal for the PorSS and that glycosylation of CTD proteins occurs after removal of the CTD region. Rabbits were used for making antisera against bacterial proteins in this study. PMID:21731719

  12. Gliding Motility and Por Secretion System Genes Are Widespread among Members of the Phylum Bacteroidetes

    PubMed Central

    Zhu, Yongtao

    2013-01-01

    The phylum Bacteroidetes is large and diverse, with rapid gliding motility and the ability to digest macromolecules associated with many genera and species. Recently, a novel protein secretion system, the Por secretion system (PorSS), was identified in two members of the phylum, the gliding bacterium Flavobacterium johnsoniae and the nonmotile oral pathogen Porphyromonas gingivalis. The components of the PorSS are not similar in sequence to those of other well-studied bacterial secretion systems. The F. johnsoniae PorSS genes are a subset of the gliding motility genes, suggesting a role for the secretion system in motility. The F. johnsoniae PorSS is needed for assembly of the gliding motility apparatus and for secretion of a chitinase, and the P. gingivalis PorSS is involved in secretion of gingipain protease virulence factors. Comparative analysis of 37 genomes of members of the phylum Bacteroidetes revealed the widespread occurrence of gliding motility genes and PorSS genes. Genes associated with other bacterial protein secretion systems were less common. The results suggest that gliding motility is more common than previously reported. Microscopic observations confirmed that organisms previously described as nonmotile, including Croceibacter atlanticus, “Gramella forsetii,” Paludibacter propionicigenes, Riemerella anatipestifer, and Robiginitalea biformata, exhibit gliding motility. Three genes (gldA, gldF, and gldG) that encode an apparent ATP-binding cassette transporter required for F. johnsoniae gliding were absent from two related gliding bacteria, suggesting that the transporter may not be central to gliding motility. PMID:23123910

  13. Oral administration of recombinant Neisseria meningitidis PorA genetically fused to H. pylori HpaA antigen increases antibody levels in mouse serum, suggesting that PorA behaves as a putative adjuvant.

    PubMed

    Vasquez, Abel E; Manzo, Ricardo A; Soto, Daniel A; Barrientos, Magaly J; Maldonado, Aurora E; Mosqueira, Macarena; Avila, Anastasia; Touma, Jorge; Bruce, Elsa; Harris, Paul R; Venegas, Alejandro

    2015-01-01

    The Neisseria meningitidis outer membrane protein PorA from a Chilean strain was purified as a recombinant protein. PorA mixed with AbISCO induced bactericidal antibodies against N. meningitidis in mice. When PorA was fused to the Helicobacter pylori HpaA antigen gene, the specific response against H. pylori protein increased. Splenocytes from PorA-immunized mice were stimulated with PorA, and an increase in the secretion of IL-4 was observed compared with that of IFN-γ. Moreover, in an immunoglobulin sub-typing analysis, a substantially higher IgG1 level was found compared with IgG2a levels, suggesting a Th2-type immune response. This study revealed a peculiar behavior of the purified recombinant PorA protein per se in the absence of AbISCO as an adjuvant. Therefore, the resistance of PorA to proteolytic enzymes, such as those in the gastrointestinal tract, was analyzed, because this is an important feature for an oral protein adjuvant. Finally, we found that PorA fused to the H. pylori HpaA antigen, when expressed in Lactococcus lactis and administered orally, could enhance the antibody response against the HpaA antigen approximately 3 fold. These observations strongly suggest that PorA behaves as an effective oral adjuvant. PMID:25750999

  14. Oral administration of recombinant Neisseria meningitidis PorA genetically fused to H. pylori HpaA antigen increases antibody levels in mouse serum, suggesting that PorA behaves as a putative adjuvant

    PubMed Central

    Vasquez, Abel E; Manzo, Ricardo A; Soto, Daniel A; Barrientos, Magaly J; Maldonado, Aurora E; Mosqueira, Macarena; Avila, Anastasia; Touma, Jorge; Bruce, Elsa; Harris, Paul R; Venegas, Alejandro

    2015-01-01

    The Neisseria meningitidis outer membrane protein PorA from a Chilean strain was purified as a recombinant protein. PorA mixed with AbISCO induced bactericidal antibodies against N. meningitidis in mice. When PorA was fused to the Helicobacter pylori HpaA antigen gene, the specific response against H. pylori protein increased. Splenocytes from PorA-immunized mice were stimulated with PorA, and an increase in the secretion of IL-4 was observed compared with that of IFN-γ. Moreover, in an immunoglobulin sub-typing analysis, a substantially higher IgG1 level was found compared with IgG2a levels, suggesting a Th2-type immune response. This study revealed a peculiar behavior of the purified recombinant PorA protein per se in the absence of AbISCO as an adjuvant. Therefore, the resistance of PorA to proteolytic enzymes, such as those in the gastrointestinal tract, was analyzed, because this is an important feature for an oral protein adjuvant. Finally, we found that PorA fused to the H. pylori HpaA antigen, when expressed in Lactococcus lactis and administered orally, could enhance the antibody response against the HpaA antigen approximately 3 fold. These observations strongly suggest that PorA behaves as an effective oral adjuvant. PMID:25750999

  15. Field-Evolved Mode 1 Resistance of the Fall Armyworm to Transgenic Cry1Fa-Expressing Corn Associated with Reduced Cry1Fa Toxin Binding and Midgut Alkaline Phosphatase Expression.

    PubMed

    Jakka, Siva R K; Gong, Liang; Hasler, James; Banerjee, Rahul; Sheets, Joel J; Narva, Kenneth; Blanco, Carlos A; Jurat-Fuentes, Juan L

    2016-02-01

    Insecticidal protein genes from the bacterium Bacillus thuringiensis (Bt) are expressed by transgenic Bt crops (Bt crops) for effective and environmentally safe pest control. The development of resistance to these insecticidal proteins is considered the most serious threat to the sustainability of Bt crops. Resistance in fall armyworm (Spodoptera frugiperda) populations from Puerto Rico to transgenic corn producing the Cry1Fa insecticidal protein resulted, for the first time in the United States, in practical resistance, and Bt corn was withdrawn from the local market. In this study, we used a field-collected Cry1Fa corn-resistant strain (456) of S. frugiperda to identify the mechanism responsible for field-evolved resistance. Binding assays detected reduced Cry1Fa, Cry1Ab, and Cry1Ac but not Cry1Ca toxin binding to midgut brush border membrane vesicles (BBMV) from the larvae of strain 456 compared to that from the larvae of a susceptible (Ben) strain. This binding phenotype is descriptive of the mode 1 type of resistance to Bt toxins. A comparison of the transcript levels for putative Cry1 toxin receptor genes identified a significant downregulation (>90%) of a membrane-bound alkaline phosphatase (ALP), which translated to reduced ALP protein levels and a 75% reduction in ALP activity in BBMV from 456 compared to that of Ben larvae. We cloned and heterologously expressed this ALP from susceptible S. frugiperda larvae and demonstrated that it specifically binds with Cry1Fa toxin. This study provides a thorough mechanistic description of field-evolved resistance to a transgenic Bt crop and supports an association between resistance and reduced Cry1Fa toxin binding and levels of a putative Cry1Fa toxin receptor, ALP, in the midguts of S. frugiperda larvae. PMID:26637593

  16. Development and feeding of fall armyworm on Miscanthus x giganteus and switchgrass.

    PubMed

    Prasifka, J R; Bradshaw, J D; Meagher, R L; Nagoshi, R N; Steffey, K L; Gray, M E

    2009-12-01

    Observations of fall armyworm, Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae), larvae infesting plots of Miscanthus x giganteus Greef and Deuter ex Hodkinson and Renvoize prompted laboratory-based tests of survival, development, and feeding preferences on leaf tissue from M. x giganteus and switchgrass, Panicum virgatum L. Survival from hatch to pupation was >70 and 50% for fall armyworms reared on switchgrass and M. x giganteus, respectively, although survival of the S. frugiperda rice strain was significantly greater than the corn strain on both crops. Developmental times from hatch to pupation or adult emergence showed effects of crop and S. frugiperda host strain, but analysis of an interaction revealed developmental times for the rice strain were similar on both crops, whereas corn strain larvae showed delayed development on M. x giganteus relative to switchgrass. Analysis of larval (10 d) and pupal masses showed a similar pattern, with effects of crop and an interaction (at 10 d), but only the mass of corn strain larvae feeding on M. x giganteus was reduced relative to the other crop and strain combinations. In choice tests, neonates of both corn and rice strains showed a strong preference for feeding on young tissues rather than mature leaves of M. x giganteus or switchgrass, but they also clearly favored corn, Zea mays L., leaves over either of the perennial grasses. Results indicate both plants are potential hosts for S. frugiperda, but additional information is needed to understand under which scenarios and to what degree fall armyworms may damage perennial grasses grown for biofuel production. PMID:20069844

  17. Oral toxicity of beta-N-acetyl hexosaminidase to insects.

    PubMed

    Dowd, Patrick F; Johnson, Eric T; Pinkerton, T Scott

    2007-05-01

    Insect chitin is a potential target for resistance plant proteins, but plant-derived chitin-degrading enzymes active against insects are virtually unknown. Commercial beta-N-acetylhexosaminidase (NAHA), a chitin-degrading enzyme from jack bean Canavalia ensiformis, caused significant mortality of fall armyworm Spodoptera frugiperda larvae at 75 microg/gm, but no significant mortality was noted with Aspergillus niger NAHA. Maize Zea mays callus transformed to express an Arabidopsis thaliana clone that putatively codes for NAHA caused significantly higher mortality of cigarette beetle Lasioderma serricorne larvae and significantly reduced growth rates (as reflected by survivor weights) of S. frugiperda as compared to callus that expressed control cDNAs. Tassels from model line Hi-II maize (Z. mays) plants transformed with the NAHA gene fed to S. frugiperda caused significantly higher mortality than tassels transformed to express glucuronidase; mortality was significantly correlated with NAHA expression levels detected histochemically. Leaf disks from inbred Oh43 maize plants transformed with the NAHA gene on average had significantly less feeding by caterpillars than null transformants. Leaf disks of Oh43 transformants caused significant mortality of both S. frugiperda and corn earworm Helicoverpa zea larvae, which was associated with higher expression levels of NAHA detected by isoelectric focusing, histochemically, or with antibody. Overall, these results suggest that plant NAHA has a role in insect resistance. Introduction of NAHA genes or enhancement of activity through breeding or genetic engineering has the potential to significantly reduce insect damage and thereby indirectly reduce mycotoxins that are harmful to animals and people. PMID:17417870

  18. Influence of infection route on the infectivity of baculovirus mutants lacking the apoptosis-inhibiting gene p35 and the adjacent gene p94.

    PubMed Central

    Clem, R J; Robson, M; Miller, L K

    1994-01-01

    The infectivity of Autographa californica nuclear polyhedrosis virus mutants lacking the apoptosis-inhibiting gene p35 is decreased 1,000-fold or more in larvae of the insect Spodoptera frugiperda if the budded form of the virus is administered by hemocoelic injection; this decrease is correlated with the antiviral effects of apoptosis (R. J. Clem and L. K. Miller, J. Virol. 67:3730-3738, 1993). We have extended this correlation by showing that the infectivity of p35 mutant budded virus is restored to wild-type levels by expression of an unrelated baculovirus apoptosis-inhibiting gene, Cp-iap. We have also examined the oral infectivity of the occluded form of mutants lacking p35, the neighboring p94 gene, or both genes by feeding insects occluded virus. The oral infectivity of the p35 mutant was significantly reduced in S. frugiperda larvae, but this reduction (25-fold) was less than that observed for the hemocoelic route of infection (1,000-fold). The disruption of p94 alone had no apparent effect on infectivity by either route. Unexpectedly, however, the disruption of both p35 and p94 restored oral infectivity to nearly wild-type levels but did not exert this compensatory effect on infectivity by hemocoelic injection. Thus, the infectivity of the double p35/p94 mutant is affected in a route-specific manner in S. frugiperda larvae, suggesting a tissue-specific response to p35 and/or p94. Infectivity in a different host, Trichoplusia ni, was unaffected by all the mutants tested, consistent with previous studies indicating a lack of sensitivity to apoptosis in this species. However, T. ni and S. frugiperda larvae infected with p35 mutants failed to exhibit the symptom of morphological disintegration ("melting") typical of a wild-type infection, suggesting that p35 is required for the infection of some tissues in both species. PMID:8084009

  19. Using field-evolved resistance to Cry1F maize in a lepidopteran pest to demonstrate no adverse effects of Cry1F on one of its major predators.

    PubMed

    Tian, Jun-Ce; Collins, Hilda L; Romeis, Jörg; Naranjo, Steven E; Hellmich, Richard L; Shelton, Anthony M

    2012-12-01

    Spodoptera frugiperda (JE Smith) represents the first documented case of field-evolved resistance to a genetically engineered crop expressing an insecticidal protein from Bacillus thuringiensis (Bt). In this case it was Cry1F-expressing maize (Mycogen 2A517). The ladybird beetle, Coleomegilla maculata, is a common and abundant predator that suppresses pest populations in maize and many other cropping systems. Its larvae and adults are polyphagous, feeding on aphids, thrips, lepidopteran eggs and larvae, as well as plant tissues. Thus, C. maculata may be exposed to Bt proteins expressed in genetically engineered crops by several pathways. Using Cry1F-resistant S. frugiperda larvae as prey, we evaluated the potential impact of Cry1F-expressing maize on several fitness parameters of C. maculata over two generations. Using Cry1F resistant prey removed any potential prey-mediated effects. Duration of larval and pupal stages, adult weight and female fecundity of C. maculata were not different when they were fed resistant S. frugiperda larvae reared on either Bt or control maize leaves during both generations. ELISA and insect-sensitive bioassays showed C. maculata were exposed to bioactive Cry1F protein. The insecticidal protein had no effect on C. maculata larvae, even though larvae contained 20-32 ng of Cry1F/g by fresh weight. Over all, our results demonstrated that the Cry1F protein did not affect important fitness parameters of one of S. frugiperda's major predators and that Cry1F protein did not accumulate but was strongly diluted when transferred during trophic interactions. PMID:22373893

  20. An Analysis of Interlanguage Development Over Time: Part 1, "por" and "para".

    ERIC Educational Resources Information Center

    Guntermann, Gail

    1992-01-01

    The first part of a larger planned investigation, this study examines the use of "por" and "para" by nine Peace Corps volunteers in oral interviews at the end of training and roughly one year later, to trace their acquisition over time, in two learning contexts. (24 references) (LB)

  1. The Acquisition of Lexical Meaning in a Study Abroad Context: The Spanish Prepositions "por" and "para."

    ERIC Educational Resources Information Center

    Lafford, Barbara A.; Ryan, John M.

    1995-01-01

    Examination of the development of form/function relations of the prepositions "por" and "para" at different levels of proficiency in the interlanguage of study-abroad students in Granada, Spain, revealed "noncanonical" as well as "canonical" uses of these prepositions. The most common noncanonical uses were as substitutions for other prepositions…

  2. The porA gene in serogroup A meningococci: evolutionary stability and mechanism of genetic variation.

    PubMed

    Suker, J; Feavers, I M; Achtman, M; Morelli, G; Wang, J F; Maiden, M C

    1994-04-01

    Molecular analyses were applied to the genes encoding variants of the serosubtyping antigen, the class 1 outer membrane protein (PorA), from 55 serogroup A Neisseria meningitidis strains. These genes were evolutionarily stable and exhibited a limited range of genetic variation, primarily generated by recombination. Translation of the gene sequences revealed a total of 19 distinct amino acid sequences in the variable regions of the protein, 6 of which were not recognized by currently available serosubtyping monoclonal antibodies. Knowledge of these amino acid sequences permitted a rational re-assignment of serosubtype names. Comparison of the complete genes with porA gene sequences from serogroup B and C meningococci showed that serogroup A possessed a limited number of the possible porA genes from a globally distributed gene pool. Each serogroup A subgroup was characterized by one of four porA gene types, probably acquired upon subgroup divergence, which was stable over periods of decades and during epidemiological spread. Comparison with other variable genes (pil and iga) indicated that the three alleles were independently assorted within the subgroup, suggesting that their gene types were older than the subgroups in which they occurred. PMID:8057850

  3. Informe a la Nación de mortalidad por cáncer sigue bajando

    Cancer.gov

    El Informe Anual a la Nación sobre el Estado del Cáncer, de 1975 a 2009, indica que los índices generales de mortalidad por cáncer siguen bajando en los Estados Unidos en hombres y mujeres, entre todos los grupos raciales y étnicos principales y para todo

  4. Detección y estudio mediante Fluorescencia Inducida por Láser de radicales libres formados por Disociación Multifotónica Infrarroja

    NASA Astrophysics Data System (ADS)

    Santos, M.; Díaz, L.; Torresano, J. A.; Rubio, L.; Samoudi, B.

    Una de las principales aplicaciones actuales de los procesos de disociación multifotónica inducidos por radiación láser infrarroja (DMI) es la producción de radiales libres, con el fin de estudiar sus propiedades cinéticas y espectroscópicas. La disociación de moléculas poliatómicas en el IR con láseres de CO2 tiene lugar desde la superficie de energía molecular mas baja y conduce generalmente a la formación de fragmentos en el estado electrónico fundamental, con diversos grados de excitación vibracional. En el Grupo de Procesos Multifotónicos del Instituto de Estructura de la Materia del C.S.I.C. hemos puesto a punto la técnica de Fluorescencia Inducida por Láser (LIF) para la detección y análisis en tiempo real de los fragmentos producidos en la DMI inducida mediante uno o dos campos láseres de diferentes longitudes de onda. Objetivos de nuestro trabajo han sido el estudio de los canales de disociación mayoritarios y de las especies transitoria producidas, así como de la distribución de energía interna con que éstas son generadas. En particular hemos detectado mediante LIF las especies: C2, CF, CH, SiH2, CF2, CH2, SiHCl, y CF3 a partir de la disociación de, entre otras, las siguientes moléculas: C2H3Br, C3F6, C4H8Si, C2H5ClSi y CH5ClSi. En este trabajo presentamos algunos de los resultados obtenidos mediante el estudio por LIF de estos radicales: estudio temporal de la señal LIF obtenida con determinación de tiempos de vida, espectros de excitación y fluorescencia, temperaturas vibracionales de formación, variación de la intensidad LIF con el tiempo de retraso entre los láseres de disociación y prueba, etc.

  5. ATP for the portable 500 CFM exhauster POR-005 skid C

    SciTech Connect

    Keller, C.M.

    1997-06-27

    This Acceptance Test Plan is for a 500 CFM Portable Exhauster POR-005 to be used for saltwell pumping. The Portable Exhauster System will be utilized to eliminate potential flammable gases that may exist within the dome space of the tank. This Acceptance Plan will test and verify that the exhauster meets the specified design criteria, safety requirements, operations requirements, and will provide a record of the functional test results.

  6. ATP for the portable 500 CFM exhauster POR-006 skid D

    SciTech Connect

    Keller, C.M.

    1997-07-29

    This Acceptance Test Plan is for a 500 CFM Portable Exhauster POR-006 to be used for saltwell pumping. The Portable Exhauster System will be utilized to eliminate potential flammable gases that may exist within the dome space of the tank. This Acceptance Plan will test and verify that the exhauster meets the specified design criteria, safety requirements, operations requirements, and will provide a record of the functional test results.

  7. ATP for the portable 500 CFM exhauster POR-004 skid B

    SciTech Connect

    Keller, C.M.

    1997-05-06

    This Acceptance Test Plan is for a 500 CFM Portable Exhauster POR-004 to be used for saltwell pumping. The Portable Exhauster System will be utilized to eliminate potential flammable gases that may exist within the dome space of the tank. This Acceptance Plan will test and verify that the exhauster meets the specified design criteria, safety requirements, operations requirements, and will provide a record of the functional test results.

  8. Correlation between Serological and Sequencing Analyses of the PorB Outer Membrane Protein in the Neisseria meningitidis Serotyping System

    PubMed Central

    Sacchi, Claudio T.; Lemos, Ana P. S.; Whitney, Anne M.; Solari, Claude A.; Brandt, Mary E.; Melles, Carmo E. A.; Frasch, Carl E.; Mayer, Leonard W.

    1998-01-01

    The current serological typing scheme for Neisseria meningitidis is not comprehensive; a proportion of isolates are not serotypeable. DNA sequence analysis and predicted amino acid sequences were used to characterize the structures of variable-region (VR) epitopes on N. meningitidis PorB proteins (PorB VR typing). Twenty-six porB gene sequences were obtained from GenBank and aligned with 41 new sequences. Primary amino acid structures predicted from those genes were grouped into 30 VR families of related variants that displayed at least 60% similarity. We correlated VR families with monoclonal antibody (MAb) reactivities, establishing a relationship between VR families and epitope locations for 15 serotype-defining MAbs. The current panel of serotype-defining MAbs underestimates by at least 50% the PorB VR variability because reagents for several major VR families are lacking or because a number of VR variants within some families are not recognized by serotype-defining MAbs. These difficulties, also reported for serosubtyping based on the PorA protein, are shown as inconsistent results between serological and sequence analyses, leading to inaccurate strain identification and incomplete epidemiological data. The information from this study enabled the expansion of the panel of MAbs currently available for serotyping, by including MAbs of previously undetermined specificities. Use of the expanded serotype panel enabled us to improve the sensitivity of serotyping by resolving a number of formerly nonserotypeable strains. In most cases, this information can be used to predict the VR family placement of unknown PorB proteins without sequencing the entire porB gene. PorB VR typing complements serotyping, and a combination of both techniques may be used for full characterization of meningococcal strains. The present work represents the most complete and integrated data set of PorB VR sequences and MAb reactivities of serogroup B and C meningococci produced to date. PMID

  9. Zinc Finger Nuclease Knock-out of NADPH:Cytochrome P450 Oxidoreductase (POR) in Human Tumor Cell Lines Demonstrates That Hypoxia-activated Prodrugs Differ in POR Dependence*

    PubMed Central

    Su, Jiechuang; Gu, Yongchuan; Pruijn, Frederik B.; Smaill, Jeff B.; Patterson, Adam V.; Guise, Christopher P.; Wilson, William R.

    2013-01-01

    Hypoxia, a ubiquitous feature of tumors, can be exploited by hypoxia-activated prodrugs (HAP) that are substrates for one-electron reduction in the absence of oxygen. NADPH:cytochrome P450 oxidoreductase (POR) is considered one of the major enzymes responsible, based on studies using purified enzyme or forced overexpression in cell lines. To examine the role of POR in HAP activation at endogenous levels of expression, POR knock-outs were generated in HCT116 and SiHa cells by targeted mutation of exon 8 using zinc finger nucleases. Absolute quantitation by proteotypic peptide mass spectrometry of DNA sequence-confirmed multiallelic mutants demonstrated expression of proteins with residual one-electron reductase activity in some clones and identified two (Hko2 from HCT116 and S2ko1 from SiHa) that were functionally null by multiple criteria. Sensitivities of the clones to 11 HAP (six nitroaromatics, three benzotriazine N-oxides, and two quinones) were compared with wild-type and POR-overexpressing cells. All except the quinones were potentiated by POR overexpression. Knocking out POR had a marked effect on antiproliferative activity of the 5-nitroquinoline SN24349 in both genetic backgrounds after anoxic exposure but little or no effect on activity of most other HAP, including the clinical stage 2-nitroimidazole mustard TH-302, dinitrobenzamide mustard PR-104A, and benzotriazine N-oxide SN30000. Clonogenic cell killing and reductive metabolism of PR-104A and SN30000 under anoxia also showed little change in the POR knock-outs. Thus, although POR expression is a potential biomarker of sensitivity to some HAP, identification of other one-electron reductases responsible for HAP activation is needed for their rational clinical development. PMID:24196959

  10. Characterization of Plasmid pOR1 from Ornithobacterium rhinotracheale and Construction of a Shuttle Plasmid

    PubMed Central

    Jansen, Ruud; Chansiripornchai, Niwat; Gaastra, Wim; van Putten, Jos P. M.

    2004-01-01

    The bacterium Ornithobacterium rhinotracheale has been recognized as an emerging pathogen in poultry since about 10 years ago. Knowledge of this bacterium and its mechanisms of virulence is still very limited. Here we report the development of a transformation system that enables genetic modification of O. rhinotracheale. The system is based on a cryptic plasmid, pOR1, that was derived from an O. rhinotracheale strain of serotype K. Sequencing indicated that the plasmid consisted of 14,787 nucleotides. Sequence analysis revealed one replication origin and several rep genes that control plasmid replication and copy number, respectively. In addition, pOR1 contains genes with similarity to a heavy-metal-transporting ATPase, a TonB-linked siderophore receptor, and a laccase. Reverse transcription-PCR demonstrated that these genes were transcribed. Other putative open reading frames exhibited similarities with a virulence-associated protein in Actinobacillus actinomycetemcomitans and a number of genes coding for proteins with unknown function. An Escherichia coli-O. rhinotracheale shuttle plasmid (pOREC1) was constructed by cloning the replication origin and rep genes from pOR1 and the cfxA gene from Bacteroides vulgatus, which codes for resistance to the antibiotic cefoxitin, into plasmid pGEM7 by using E. coli as a host. pOREC1 was electroporated into O. rhinotracheale and yielded cefoxitin-resistant transformants. The pOREC1 isolated from these transformants was reintroduced into E. coli, demonstrating that pOREC1 acts as an independent replicon in both E. coli and O. rhinotracheale, fulfilling the criteria for a shuttle plasmid that can be used for transformation, targeted mutagenesis, and the construction of defined attenuated vaccine strains. PMID:15466524

  11. Expression, purification and preliminary X-ray analysis of the Neisseria meningitidis outer membrane protein PorB

    SciTech Connect

    Tanabe, Mikio; Iverson, Tina M.

    2010-01-28

    The Neisseria meningitidis outer membrane protein PorB was expressed in Escherichia coli and purified from inclusion bodies by denaturation in urea followed by refolding in buffered LDAO on a size-exclusion column. PorB has been crystallized in three different crystal forms: C222, R32 and P6{sub 3}. The C222 crystal form may contain either one or two PorB monomers in the asymmetric unit, while both the R32 and P6{sub 3} crystal forms contained one PorB monomer in the asymmetric unit. Of the three, the P6{sub 3} crystal form had the best diffraction quality, yielding data extending to 2.3 {angstrom} resolution.

  12. Application of POR-Tveks to the radiochemical recovery of yttrium-90

    SciTech Connect

    Maksimova, A.M.; Kvasnitskii, I.B.

    1988-01-01

    The authors describe a method for the radiochemical analysis of fish bones for the accumulation of strontium 90 and yttrium 90 from power plant contamination of surface waters which involves labelling the sample with isotopes and subsequent adsorption of the yttrium component with the use of POR-Tveks, an adsorbent based on a copolymer of styrene and divinylbenzene with heteroradical phosphine oxide. The yield of yttrium is determined from the mass of the oxide and from the half-life of the yttrium isotope.

  13. Antigenic topology of chlamydial PorB protein and identification of targets for immune neutralization of infectivity.

    PubMed

    Kawa, Diane E; Stephens, Richard S

    2002-05-15

    The outer membrane protein PorB is a conserved chlamydial protein that functions as a porin and is capable of eliciting neutralizing Abs. A topological antigenic map was developed using overlapping synthetic peptides representing the Chlamydia trachomatis PorB sequence and polyclonal immune sera. To identify which antigenic determinants were surface accessible, monospecific antisera were raised to the PorB peptides and were used in dot-blot and ELISA-based absorption studies with viable chlamydial elementary bodies (EBs). The ability of the surface-accessible antigenic determinants to direct neutralizing Ab responses was investigated using standardized in vitro neutralization assays. Four major antigenic clusters corresponding to Phe(34)-Leu(59) (B1-2 and B1-3), Asp(112) -Glu(145) (B2-3 and B2-4), Gly(179)-Ala(225) (B3-2 to B3-4), and Val(261)-Asn(305) (B4-4 to B5-2) were identified. Collectively, the EB absorption and dot-blot assays established that the immunoreactive PorB Ags were exposed on the surface of chlamydial EBs. Peptide-specific antisera raised to the surface-accessible Ags neutralized chlamydial infectivity and demonstrated cross-reactivity to synthetic peptides representing analogous C. pneumoniae PorB sequences. Furthermore, neutralization of chlamydial infectivity by C. trachomatis PorB antisera was inhibited by synthetic peptides representing the surface-exposed PorB antigenic determinants. These findings demonstrate that PorB Ags may be useful for development of chlamydial vaccines. PMID:11994474

  14. Multiple active site residues are important for photochemical efficiency in the light-activated enzyme protochlorophyllide oxidoreductase (POR).

    PubMed

    Menon, Binuraj R K; Hardman, Samantha J O; Scrutton, Nigel S; Heyes, Derren J

    2016-08-01

    Protochlorophyllide oxidoreductase (POR) catalyzes the light-driven reduction of protochlorophyllide (Pchlide), an essential, regulatory step in chlorophyll biosynthesis. The unique requirement of the enzyme for light has provided the opportunity to investigate how light energy can be harnessed to power biological catalysis and enzyme dynamics. Excited state interactions between the Pchlide molecule and the protein are known to drive the subsequent reaction chemistry. However, the structural features of POR and active site residues that are important for photochemistry and catalysis are currently unknown, because there is no crystal structure for POR. Here, we have used static and time-resolved spectroscopic measurements of a number of active site variants to study the role of a number of residues, which are located in the proposed NADPH/Pchlide binding site based on previous homology models, in the reaction mechanism of POR. Our findings, which are interpreted in the context of a new improved structural model, have identified several residues that are predicted to interact with the coenzyme or substrate. Several of the POR variants have a profound effect on the photochemistry, suggesting that multiple residues are important in stabilizing the excited state required for catalysis. Our work offers insight into how the POR active site geometry is finely tuned by multiple active site residues to support enzyme-mediated photochemistry and reduction of Pchlide, both of which are crucial to the existence of life on Earth. PMID:27285815

  15. Diagnóstico diferencial en la encefalitis por anticuerpos contra el receptor NMDA

    PubMed Central

    González-Valcárcel, J.; Rosenfeld, M.R.; Dalmau, J.

    2011-01-01

    Resumen Introducción La encefalitis por anticuerpos contra el receptor de NMDA (NMDAR) suele desarrollarse como un síndrome característico de evolución multifásica y diagnóstico diferencial amplio. Pacientes Presentamos a 2 pacientes diagnosticadas de encefalitis por anticuerpos NMDAR con un cuadro clínico típico, pero que inicialmente señaló otras etiologías. Discusión La afectación frecuente de pacientes jóvenes con manifestaciones psiquiátricas prominentes indica frecuentemente otras consideraciones diagnósticas; las más frecuentes son las encefalitis virales, los procesos psiquiátricos y el síndrome neuroléptico maligno. Varios síndromes previamente definidos de manera parcial o descriptiva en adultos y pacientes pediátricos probablemente eran casos de encefalitis anti-NMDAR. Conclusiones La encefalitis anti-NMDAR debe considerarse en pacientes jóvenes con manifestaciones psiquiátricas subagudas, movimientos anormales y alteraciones autonómicas. La caracterización clínica e inmunológica de esta enfermedad ha llevado a la identificación de nuevos anticuerpos que afectan a procesos de memoria, aprendizaje, conducta y psicosis. PMID:20964986

  16. Effect of SPM-based cleaning POR on EUV mask performance

    NASA Astrophysics Data System (ADS)

    Choi, Jaehyuck; Lee, Han-shin; Yoon, Jinsang; Shimomura, Takeya; Friz, Alex; Montgomery, Cecilia; Ma, Andy; Goodwin, Frank; Kang, Daehyuk; Chung, Paul; Shin, Inkyun; Cho, H.

    2011-11-01

    EUV masks include many different layers of various materials rarely used in optical masks, and each layer of material has a particular role in enhancing the performance of EUV lithography. Therefore, it is crucial to understand how the mask quality and patterning performance can change during mask fabrication, EUV exposure, maintenance cleaning, shipping, or storage. The fact that a pellicle is not used to protect the mask surface in EUV lithography suggests that EUV masks may have to undergo more cleaning cycles during their lifetime. More frequent cleaning, combined with the adoption of new materials for EUV masks, necessitates that mask manufacturers closely examine the performance change of EUV masks during cleaning process. We have investigated EUV mask quality and patterning performance during 30 cycles of Samsung's EUV mask SPM-based cleaning and 20 cycles of SEMATECH ADT exposure. We have observed that the quality and patterning performance of EUV masks does not significantly change during these processes except mask pattern CD change. To resolve this issue, we have developed an acid-free cleaning POR and substantially improved EUV mask film loss compared to the SPM-based cleaning POR.

  17. Methodology for obtaining stakeholder assessments of obesity policy options in the PorGrow project.

    PubMed

    Stirling, A; Lobstein, T; Millstone, E

    2007-05-01

    The Policy Options for Responding to the Growing Challenge of Obesity Research Project (PorGrow) study provided a unique opportunity to develop a large-scale application of a semi-quantitative technique for exploring interviewees' views on options to tackle obesity, using multi-criteria mapping. This 'heuristic' approach utilizes the advantages of a structured interviews framework by predefining a set of options for appraisal, while leaving interviewees free to select their own criteria for making their judgements. Additional information can be gleaned from the interview transcripts and related materials to set the appraisals in their policy context, and allowing interviewees to express their views on the options presented and their own appraisals. The PorGrow study team agreed a predefined set of 20 options for appraisal, and interviewed sets of stakeholders representing more than 20 aspects of policy development in each of the nine participating countries. The details of the methodology adopted are set out in this paper. PMID:17371304

  18. Geometry sensing through POR1 regulates Rac1 activity controlling early osteoblast differentiation in response to nanofiber diameter.

    PubMed

    Higgins, A M; Banik, B L; Brown, J L

    2015-02-01

    Bone grafting procedures in the United States rely heavily upon autografts and allografts, which are donor-dependent, cause donor site pain, and can transmit disease. Synthetic bone grafts can reduce these risks; however, synthetics lack the bone differentiating (osteoinductive) abilities of auto- and allografts. Achieving innate osteoinductive properties of synthetics through surface modifications is currently under investigation. This study focuses on nanofibers, with emphasis on how fiber diameter and the potential curvature sensor POR1 affect the activation of the signaling molecules Rac1 and Arf1, and leading to expression of alkaline phosphatase (ALP), an osteoinductive marker. Diameters of 0.1, 0.3, and 1.0 μm were compared against a flat control. The highest level of Rac1 activation was achieved on the smallest fibers (0.1 μm), a trend that was lost in POR1 knockdowns. This supports the hypothesis that on small nanofibers, POR1 favorably binds to highly curved cell membranes, which allows Rac1 to subsequently dissociate and activate. When the curvature is insufficient to bind POR1, POR1 binds to inactive Rac1 and competitively inhibits its activation. Arf1 activation followed an opposite trend, with the largest nanofibers exhibiting the highest activity. This trend reinforces the known interaction between Rac1 and Arf1 through the GIT-PIX complex, an Arf1 GAP and Rac1 GEF, respectively. Large, (1.0 μm), nanofibers demonstrated the highest ALP activity, indicating that ALP expression is inversely dependent on Rac1 activation. Knockdown of POR1 resulted in increased ALP activity across the substrates but without regard to the curvature sensing trend seen previously. Thus, POR1 senses curvature and increases Rac1 activity, which negatively regulates bone differentiation. PMID:25539497

  19. Substrate-specific modulation of CYP3A4 activity by genetic variants of cytochrome P450 oxidoreductase (POR)

    PubMed Central

    Agrawal, Vishal; Choi, Ji Ha; Giacomini, Kathleen M.; Miller, Walter L.

    2010-01-01

    Objectives CYP3A4 receives electrons from P450 oxidoreductase (POR) to metabolize about 50% of clinically used drugs. There is substantial inter-individual variation in CYP3A4 catalytic activity that is not explained by CYP3A4 genetic variants. CYP3A4 is flexible and distensible, permitting it to accommodate substrates varying in shape and size. To elucidate mechanisms of variability in CYP3A4 catalysis, we examined the effects of genetic variants of POR, and explored the possibility that substrate-induced conformational changes in CYP3A4 differentially affect the ability of POR variants to support catalysis. Methods We expressed human CYP3A4 and four POR variants (Q153R, A287P, R457H, A503V) in bacteria, reconstituted them in vitro and measured the Michaelis constant and maximum velocity with testosterone, midazolam, quinidine and erythromycin as substrates. Results POR A287P and R457H had low activity with all substrates; Q153R had 76–94% of wild type (WT) activity with midazolam and erythromycin, but 129–150% activity with testosterone and quinidine. The A503V polymorphism reduced CYP3A4 activity to 61–77% of wild type with testosterone and midazolam, but had nearly wild type activity with quinidine and erythromycin. Conclusion POR variants affect CYP3A4 activities. The impact of a POR variant on catalysis by CYP3A4 is substrate-specific, probably due to substrate-induced conformational changes in CYP3A4. PMID:20697309

  20. Intermitência alfvênica gerada por caos na atmosfera solar e no vento solar

    NASA Astrophysics Data System (ADS)

    Rempel, E. L.; Chian, A. C.-L.; Macau, E. E. N.; Rosa, R. R.

    2003-08-01

    Dados medidos no vento solar rápido proveniente dos buracos coronais revelam que os plasmas no meio interplanetário são dominados por flutuações Alfvênicas, caracterizadas por uma alta correlação entre as variações do campo magnético e da velocidade do plasma. As flutuações exibem muitas características esperadas em turbulência magneto-hidrodinâmica totalmente desenvolvida, tais como intermitência e espectros contínuos. Contudo, os mecanismos responsáveis pela evolução de turbulência Alfvênica intermitente não são completamente compreendidos. Neste trabalho a teoria de caos é usada para explicar como sistemas Alfvênicos, modelados pela equação Schrödinger não-linear derivativa e pela equação Kuramoto-Sivashinsky, podem se tornar fortemente caóticos à medida em que parâmetros do plasma são variados. Pequenas perturbações no parâmetro de dissipação podem fazer com que o sistema mude bruscamente de um regime periódico, ou fracamente caótico, para um regime fortemente caótico. As séries temporais das flutuações do campo magnético nos regimes fortemente caóticos exibem comportamento intermitente, em que fases laminares ou fracamente caóticas são interrompidas por fortes estouros caóticos. É mostrado que o regime fortemente caótico é atingido quando as soluções periódicas ou fracamente caóticas globalmente estáveis interagem com soluções do sistema que são fortemente caóticas, mas globalmente instáveis. Estas soluções globalmente instáveis são conjuntos caóticos não-atrativos conhecidos como selas caóticas, e são responsáveis pelos fortes estouros nos regimes intermitentes. Selas caóticas têm sido detectadas experimentalmente em uma grande variedade de sistemas, sendo provável que elas desempenhem um papel importante na turbulência intermitente observada em plasmas espaciais.