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Differential crop damage by healthy and nucleopolyhedrovirus-infected Mamestra brassicae L. (Lepidoptera: Noctuidae) larvae: a field examination.

Baculovirus infection in Lepidoptera can alter both larval mobility and feeding rates, which can in turn affect pathogen transmission and dispersal in the field. We compared the damage to cabbage plants in the field caused by healthy and nucleopolyhedrovirus-infected Mamestra brassicae L. (Lepidoptera: Noctuidae) larvae released as second and fourth instars. There was no significant difference in plant consumption by healthy and infected larvae for the first 4 days after release. From day 5 onwards, infected larvae caused significantly less defoliation. This pattern was similar for larvae at both larval instars. Defoliation was greater for fourth instars throughout the experiment.

Animals↗

New cell lines from larval fat bodies of Spodoptera exigua: characterization and susceptibility to baculoviruses (Lepidoptera: Noctuidae).

Two new cell lines, designated IOZCAS-Spex-II and IOZCAS-Spex-III, were initiated from the fat bodies of larvae of the beet armyworm, Spodoptera exigua (Lepidoptera: Noctuidae) in TNM-FH medium containing 10% fetal bovine serum. The spherical cells were predominant among the various cell types and measures approximately 15 microm in diameter. The cell lines were mainly composed of tetraploid cells with chromosome numbers ranging from 116 to 131 (n=31). The cell lines were confirmed to have originated from the S. exigua by DAF-PCR technique. They were susceptible to the multiple nucleocapsid nuclear polyhedrosis viruses from S. exigua.

Animals↗

Antennal lobe projection patterns of olfactory receptor neurons involved in sex pheromone detection in Spodoptera littoralis (Lepidoptera: Noctuidae).

Pheromone-specific receptor neurons in male and female cotton leafworms, Spodoptera littoralis (Boisd.) (Lepidoptera: Noctuidae) were typed physiologically and traced into the antennal lobe using cobalt lysine as a marker. In male S. littoralis, the macroglomerular complex (MGC), which is responsible for the initial integration of information concerning sex pheromone components, contains three glomerular compartments as revealed in a morphological study. No such specialized structure was seen in the female. In the male, olfactory receptor neurons that responded selectively to stimulation with the major sex pheromone component, (Z)9, (E)11-tetradecadienyl acetate (Z9E11-14:OAc), had arborizations that were restricted to a large glomerulus of the MGC (designated a), situated near the antennal nerve entrance into the antennal lobe. Neurons that were stimulated by (Z)9,(E)12-tetradecadienyl acetate (Z9E12-14:OAc), a second pheromone component, had arborizations in a lateral, smaller glomerulus of the MGC (designated c), while receptor neurons specifically tuned to a behavioural antagonist, (Z)-9-tetradecenol (Z9-14:OH), projected to a medial glomerulus of the MGC (b). In the female, receptor neurons tuned to the major pheromone component projected to a glomerulus situated at the entrance of the antennal nerve. This glomerulus did, however, not have the size or the structure of the male MGC. A second neuron housed in the same sensillum projected its axon to an ordinary glomerulus situated medially in the antennal lobe.

Animals↗

The role of allatostatic and allatotropic neuropeptides in the regulation of juvenile hormone biosynthesis in Lacanobia oleracea (Lepidoptera: Noctuidae).

In the sphinghid moth Manduca sexta, two allatoactive neuropeptides appear to be responsible for regulating juvenile hormone (JH) production by the corpora allata (CA). These peptides (M. sexta allatostatin, Mas-AS, and M. sexta allatotropin, Mas-AT) respectively inhibit and stimulate in vitro JH biosynthesis by CA in this insect. However, although Mas-AS inhibits CA in both larval and adult insects, Mas-AT is active only in adult M. sexta. The situation in other lepidopteran species is less clear-cut and, although both peptides have been detected (usually by immunologic and/or molecular techniques) in several other moths (including noctuids), their function as regulators of JH production remains uncertain. In the tomato moth Lacanobia oleracea (Lepidoptera: Noctuidae), we have previously demonstrated the occurrence of Mas-AS and/or Mas-AT in extracts of CA, brain and other organs, and have shown that both peptides are present in larval and adult forms. However, in L. oleracea, although Mas-AS inhibits larval and adult CA in vitro, it does so only at relatively high concentrations, and to a maximum of only approximately 70%. By contrast, Mas-AT (which is also present in larval and adult L. oleracea) stimulates larval and adult CA, but is substantially more potent ( approximately 100 fold) than the allatostatin. In this paper we present the results of paired, concurrent measurements (using ELISA) of levels of Mas-AS and Mas-AT in brains, CA and hemolymph (plasma and hemocytes) of L. oleracea at times when there are marked changes in JH titers. We also present data on the in vitro rates of JH biosynthesis by isolated CA, and on hemolymph JH esterase activity measured at the same critical developmental times, and discuss all of these data in relation to the putative allatoregulatory roles of the M. sexta allatotropic and allatostatic neuropeptides in L. oleracea.

Animals↗

Antifeedant effects of the limonoids from Entandrophragma candolei (Meliaceae) on the gram pod borer, Helicoverpa armigera (Lepidoptera: Noctuidae).

The biological activity of the limonoids prieurianin and epoxyprieurianin isolated from Entandrophragma candolei (Harms) (Meliaceae) and their respective acetates was assessed using the gram pod borer, Helicoverpa armigera (Hubner) (Lepidoptera: Noctuidae). The compounds exhibited strong antifeedant activity in a diet choice bioassay with epoxyprieurianin acetate being most effective with 48.3 ppm deterring feeding by 50% (DI50) and prieurianin the least effective (DI50 = 91.4 ppm). The effect on growth of larvae was concomitant with the reduced feeding by neonate and third instar larvae. In nutritional assays, all the compounds reduced growth and consumption when fed to larvae without any effect on efficiency of conversion of ingested food (ECI), suggesting antifeedant activity alone. No toxicity was observed nor was there any significant affect on nutritional indices following topical application, further suggesting that prieurianin-type limonoids act specifically as feeding deterrents.

Animals↗

Metabolism of L-canavanine and L-canaline in the tobacco budworm, Heliothis virescens [Noctuidae].

The metabolism of L-canavanine and L-canaline were investigated in larvae of the tobacco budworm, Heliothis virescens [Noctuidae]. H. virescens larvae were treated with L-[1,2,3,4-14C]canavanine or L-[U-14C]canaline with sufficient cold carrier to provide 5 mg g-1 canavanine or a molar equivalent of canaline (3.81 mg g-1). The preponderant catabolite in both canavanine- and canaline-treated larvae was [14C]homoserine. Other minor metabolites derived from canavanine included [14C]aspartate/asparagine, [14C]glutamate/glutamine, [14C]2-aminobutyrate, [14C]ornithine, [14C]proline, and [14C]isoleucine. Canaline yielded [14C]glutamate/glutamine, [14C]aspartate/asparagine, and [14C]-2-aminobutyrate. Our current studies support the belief that this destructive insect tolerates L-canavanine and L-canaline because of its ability to reductively cleave these potentially insecticidal natural products to L-homoserine and guanidine or ammonia, respectively.

Aminobutyrates↗

Adaptation of Spodoptera exigua (Lepidoptera: Noctuidae) to barley trypsin inhibitor BTI-CMe expressed in transgenic tobacco.

Nicotiana tabacum plants were transformed with the cDNA of barley trypsin inhibitor BTI-CMe under the control of the 35S CaMV promoter. Although the transgene was expressed and the protein was active in the homozygous lines selected, growth of Spodoptera exigua (Lepidoptera: Noctuidae) larvae reared on transgenic plants was not affected. The protease activity in larval midgut extracts after 2 days feeding on transformed tobacco leaves from the highest expressing plant showed a reduction of 25% in the trypsin-like activity compared to that from insects fed on non-transformed controls. The susceptibility of digestive serine-proteases to inhibition by BTI-CMe was confirmed by activity staining gels. This decrease was compensated with a significant induction of leucine aminopeptidase-like and carboxipeptidase A-like activities, while chymotrypsin-, elastase-, and carboxipeptidase B-like proteases were not affected.

Adaptation, Physiological↗

The biochemical basis for L-canavanine tolerance by the tobacco budworm Heliothis virescens (Noctuidae).

The tobacco budworm, Heliothis virescens (Noctuidae), a destructive insect pest, is remarkably resistant to L-canavanine, L-2-amino-4-(guanidinooxy)butyric acid, an arginine antimetabolite that is a potent insecticide for nonadapted species. H. virescens employs a constitutive enzyme of the larval gut, known trivially as canavanine hydrolase (CH), to catalyze an irreversible hydrolysis of L-canavanine to L-homoserine and hydroxyguanidine. As such, it represents a new type of hydrolase, one acting on oxygen-nitrogen bonds (EC 3.13.1.1). This enzyme has been isolated from the excised gut of H. virescens and purified to homogeneity; it exhibits an apparent Km value for L-canavanine of 1.1 mM and a turnover number of 21.1 micromol x min(-1)x micromol(-1). This enzyme has a mass of 285 kDa and is composed of two subunits with a mass of 50 kDa or 47.5 kDa. CH has a high degree of specificity for L-canavanine as it cannot function effectively with either L-2-amino-5-(guanidinooxy)pentanoate or L-2-amino-3-(guanidinooxy)propionate, the higher or lower homolog of L-canavanine, respectively. L-Canavanine derivatives such as methyl-L-canavanine, or L-canaline and O-ureido-L-homoserine, are not metabolized significantly by CH.

Acetone↗

Selection of a nucleopolyhedrovirus for control of Spodoptera frugiperda (Lepidoptera: Noctuidae): structural, genetic, and biological comparison of four isolates from the Americas.

Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae) is the principal pest of maize in tropical and subtropical regions of the Americas. Larvae of this species are susceptible to a nucleopolyhedrovirus (NPV) which has attracted interest as a potential biocontrol agent. Four strains of NPV isolated from infected S. frugiperda larvae in the United States, Nicaragua, and Argentina were subjected to a structural, genetic, and biological comparison to select a candidate isolate for use in biocontrol experiments in Mexico and Honduras. All isolates had an occlusion body polyhedrin protein of 32 kDa, but the virions of each isolate differed subtly in the pattern and abundance of certain structural polypeptides revealed by SDS-PAGE analysis. Restriction endonuclease analysis of viral DNA confirmed that these isolates were strains of a single virus species but showed that they were not genetically homogeneous; each isolate could be differentiated from the others using common restriction enzymes. Droplet feeding bioassays indicated that an isolate from Nicaragua (Sf-NIC) and an isolate from the United States (Sf-US) had the highest infectivity when tested against 2nd instars originating from a Honduran S. frugiperda colony. No significant differences were detected in the speed of kill of Sf-NIC (102.7 h), Sf-US (102.3 h) and Sf-AR (103.4 h), whereas that of Sf-2 (97.3 h) was significantly shorter. Additional bioassays of the Sf-NIC isolate against 2nd to 6th instars demonstrated that LC50 values increased with larval stage from 2.03 x 10(5) OBs/ml for 2nd instars to 1.84 x 10(8) OBs/ml for 5th instars. The concentration required to elicit a lethal infection of 6th instars was so high that a reliable estimate of LC50 could not be obtained. The mean time to death for each stage challenged with the Sf-NIC isolate increased with instar from an average of 102.7 h in 2nd instars to 136.9 h in 5th instars.

Analysis of Variance↗

Feeding and maturation by soybean looper (Lepidoptera: Noctuidae) larvae on soybean affected by weed, fungus, and nematode pests.

Feeding and maturation by the soybean looper, Pseudoplusia includens (Walker) (Lepidoptera: Noctuidae), were investigated in a 2-yr study on 'Davis' soybean, Glycine max (L.) Merr., grown alone and combined with the weed hemp sesbania, Sesbania exaltata (Raf.) Rybd. ex. A. W. Hill, the root-knot nematode, Meloidogyne incognita (Kofoid & White) Chitwood, and the charcoal rot fungus, Macrophomina phaseolina (Tassi) Goid. Of the three pests, hemp sesbania had the greatest effects on plant growth and insect feeding and maturation. When fed foliage from soybean stressed by hemp sesbania, soybean looper larvae remained longer in feeding stages, consumed more foliage, and showed altered weight gain compared with larvae fed control foliage. Results suggest that nutrient (s) critical for proper development of larvae may have been limited in weed-stressed soybean foliage. Less dramatic results were observed when larvae fed on foliage from soybean with roots colonized by the charcoal rot fungus. Such larvae consumed more foliage, weighed more, and showed a slight increase in larval feeding period, but only in 1 yr of the study. Colonization of soybean roots by the root-knot nematode had no consistent effects on either the soybean host or insect.

Animals↗

Phenology of Lacanobia subjuncta (Lepidoptera: Noctuidae) in Washington and Oregon apple orchards.

The phenology of Lacanobia subjuncta (Grote & Robinson) (Lepidoptera: Noctuidae) was investigated in 30 apple orchards in central Washington state and northeastern Oregon from 1998 to 2001 (57 total orchard-yr). Adult captures in pheromone-baited traps were fit to a Weibull distribution to model emergence of the first and second generations. Initial capture of first generation adults was observed at 216.2 +/- 2.6 degree-days (DD) (mean +/- SEM) from 1 March by using a base temperature of 6.7 degrees C. The model predicted that flight was 5 and 95% complete by 240 and 700 degree-days (DD), respectively. Monitoring of oviposition and hatch was used to establish a protandry plus preoviposition degree-day requirement of 160.0 +/- 7.7 DD, as well as to provide data to describe the entire hatch period. Egg hatch was 5 and 95% complete by 395 and 630 DD, respectively. The start of the second flight was observed at 1217.1 +/- 8.3 DD by using an upper threshold for development of 32 degrees C and a horizontal cutoff. The model indicated that the second flight was 5 and 95% complete by 1220 and 1690 DD, respectively. Second generation hatch was 5 and 95% complete by 1440 and 1740 DD, respectively. A discussion of the potential uses of these detailed phenology data in optimizing management strategies is presented.

Agriculture↗

Population dynamics and gene flow of Helicoverpa armigera (Lepidoptera: Noctuidae) on cotton and grain crops in the Murrumbidgee Valley, Australia.

The population dynamics of Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) in the Murrumbidgee Valley, Australia, has been characterized using five highly variable microsatellite loci. In the 2001-2002 growing season, there were very high levels of migration into the Murrumbidgee Valley with no detectable genetic structuring, consistent with previous analyses on a national scale. By contrast, there was significant genetic structuring over the 2002-2003 growing season, with three distinct genetic types detected. The first type corresponded to the first two generations and was derived from local individuals emerging from diapause and their progeny. The second genetic type corresponded to generation 3 and resulted from substantial immigration into the region. There was another genetic shift in generation 4, which accounts for the third genetic type of the season. This genetic shift occurred despite low levels of immigration. During the third generation of the 2002-2003 growing season, different population dynamics was characterized for H. armigera on maize, Zea mays L., and cotton Gossipium hirsutum L. Populations on cotton tended to cycle independently with very little immigration from outside the region or from maize within the region. Maize acted as a major sink for immigrants from cotton and from outside the region. If resistance were to develop on cotton under these circumstances, susceptible individuals from maize or from other regions would not dilute this resistance. In addition, resistance is likely to be transferred to maize and be perpetuated until diapause, from where it may reemerge next season. If low levels of immigration were to occur on transgenic cotton, this may undermine the effectiveness of refugia, especially noncotton refugia.

Animals↗

Bacterially produced dsRNA targeting SePGRP-LB reduces population fitness of Spodoptera exigua (Lepidoptera: Noctuidae) and increases its susceptibility to SeMNPV.

The beet armyworm, Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae), is an important agricultural pest, and S. exigua multiple nucleopolyhedrovirus (SeMNPV) is a host-specific biological control agent. However, baculovirus efficacy can be limited by host antiviral responses. S. exigua peptidoglycan recognition protein LB (SePGRP-LB) has been identified as an antiviral immune factor, suggesting that its suppression may increase larval susceptibility to SeMNPV. In this study, bacterially produced double-stranded RNA targeting SePGRP-LB (bac-dsPGRP-LB) was orally delivered to larvae to induce RNA interference. Feeding bac-dsPGRP-LB reduced SePGRP-LB transcript levels by 24.0% to 65.7% over 7 d. SePGRP-LB knockdown prolonged fifth-instar larval development, reduced female pupal weight, shortened male adult longevity and the oviposition period, and decreased fecundity by approximately 51%. Life table analysis further showed significant reductions in the intrinsic rate of increase (r), finite rate of increase (λ), and net reproductive rate (R0) following bac-dsPGRP-LB treatment. During SeMNPV infection, co-feeding with bac-dsPGRP-LB significantly suppressed SePGRP-LB expression, increased the SeMNPV genomic load, and reduced larval survival compared with the SeMNPV + bac-dsGFP treatment. These findings identify SePGRP-LB as a promising RNAi target for simultaneously reducing S. exigua fitness and enhancing its susceptibility to SeMNPV under laboratory conditions.

SePGRP-LB↗

Molecular and insecticidal characterization of a Cry1I protein toxic to insects of the families Noctuidae, Tortricidae, Plutellidae, and Chrysomelidae.

The most notable characteristic of Bacillus thuringiensis is its ability to produce insecticidal proteins. More than 300 different proteins have been described with specific activity against insect species. We report the molecular and insecticidal characterization of a novel cry gene encoding a protein of the Cry1I group with toxic activity towards insects of the families Noctuidae, Tortricidae, Plutellidae, and Chrysomelidae. PCR analysis detected a DNA sequence with an open reading frame of 2.2 kb which encodes a protein with a molecular mass of 80.9 kDa. Trypsin digestion of this protein resulted in a fragment of ca. 60 kDa, typical of activated Cry1 proteins. The deduced sequence of the protein has homologies of 96.1% with Cry1Ia1, 92.8% with Cry1Ib1, and 89.6% with Cry1Ic1. According to the Cry protein classification criteria, this protein was named Cry1Ia7. The expression of the gene in Escherichia coli resulted in a protein that was water soluble and toxic to several insect species. The 50% lethal concentrations for larvae of Earias insulana, Lobesia botrana, Plutella xylostella, and Leptinotarsa decemlineata were 21.1, 8.6, 12.3, and 10.0 microg/ml, respectively. Binding assays with biotinylated toxins to E. insulana and L. botrana midgut membrane vesicles revealed that Cry1Ia7 does not share binding sites with Cry1Ab or Cry1Ac proteins, which are commonly present in B. thuringiensis-treated crops and commercial B. thuringiensis-based bioinsecticides. We discuss the potential of Cry1Ia7 as an active ingredient which can be used in combination with Cry1Ab or Cry1Ac in pest control and the management of resistance to B. thuringiensis toxins.

Amino Acid Sequence↗

A Bacillus thuringiensis insecticidal crystal protein with a high activity against members of the family Noctuidae.

The full characterization of a novel insecticidal crystal protein, named Cry9Ca1 according to the revised nomenclature for Cry proteins, from Bacillus thuringiensis serovar tolworthi is reported. The crystal protein has 1,157 amino acids and a molecular mass of 129.8 kDa. It has the typical features of the Lepidoptera-active crystal proteins such as five conserved sequence blocks. Also, it is truncated upon trypsin digestion to a toxic fragment of 68.7 kDa by removal of 43 amino acids at the N terminus and the complete C-terminal half after conserved sequence block 5. The 68.7-kDa fragment is further degraded to a nontoxic 55-kDa fragment. The crystal protein has a fairly broad spectrum of activity against lepidopteran insects, including members of the families Pyralidae, Plutellidae, Sphingidae, and Noctuidae. A 50% lethal concentration of less than 100 ng/cm2 of diet agar was found for diamondback moth, European corn borer, cotton bollworm, and beet armyworm. It is the first insecticidal crystal protein with activity against cutworms. No activity was observed against some beetles, such as Colorado potato beetle. The protein recognizes a receptor different from that recognized by Cry1Ab5 in Ostrinia nubilalis and Plutella xylostella. In Spodoptera exigua and P. xylostella, it binds to a receptor which is also recognized by Cry1Cax but with a lower affinity. In these insects, Cry1Cax probably binds with a higher affinity to an additional receptor which is not recognized by Cry9Ca1. Elimination of a trypsin cleavage site which is responsible for the degradation to a nontoxic fragment did result in protease resistance but not in increased toxicity against O. nubilalis.

Amino Acid Sequence↗

The genome sequence of the Treble Lines, Charanyca trigrammica (Hufnagel, 1766) (Lepidoptera: Noctuidae).

We present a genome assembly from an individual male Charanyca trigrammica (Treble Lines; Arthropoda; Insecta; Lepidoptera; Noctuidae). The assembly contains two haplotypes with total lengths of 546.43 megabases and 546.58 megabases. Most of haplotype 1 (99.97%) is scaffolded into 31 chromosomal pseudomolecules, including the Z sex chromosome. Haplotype 2 was assembled to scaffold level. The mitochondrial genome has also been assembled, with a length of 15.44 kilobases. This assembly was generated as part of the Darwin Tree of Life project, which produces reference genomes for eukaryotic species found in Britain and Ireland.

Charanyca trigrammica; Treble Lines; genome sequen↗

The genome sequence of the Golden Plusia, Polychrysia moneta (Fabricius, 1787) (Lepidoptera: Noctuidae).

We present a genome assembly from a male specimen of Polychrysia moneta (Golden Plusia; Arthropoda; Insecta; Lepidoptera; Noctuidae). The assembly contains two haplotypes with total lengths of 430.64 megabases and 425.60 megabases. Most of haplotype 1 (98.46%) is scaffolded into 31 chromosomal pseudomolecules, including the Z sex chromosome. Haplotype 2 was assembled to scaffold level. The mitochondrial genome has also been assembled, with a length of 15.26 kilobases. This work is part of Project Psyche, a collaborative programme generating genomes for European butterflies and moths.

Polychrysia moneta; Golden Plusia; genome sequence↗

The genome sequence of the Common Quaker, Orthosia cerasi (Fabricius, 1775) (Lepidoptera: Noctuidae).

We present a genome assembly from an individual female Orthosia cerasi (Common Quaker; Arthropoda; Insecta; Lepidoptera; Noctuidae). The genome sequence has a total length of 1 022.21 megabases. Most of the assembly (99.88%) is scaffolded into 29 chromosomal pseudomolecules, including the Z sex chromosome. The mitochondrial genome has also been assembled, with a length of 15.39 kilobases. Gene annotation of this assembly on Ensembl identified 15 702 protein-coding genes. This assembly was generated as part of the Darwin Tree of Life project, which produces genomes for eukaryotic species found in Britain and Ireland.

Common Quaker↗