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Microbiome research is currently gaining tremendous interest on the impact of diets and environment to influence life traits of several hosts. Full understanding of the insect ecology and characterization of gut microbial community including their natural enemies is vital for development of novel insect pest management strategies. This study examined the gut bacterial communities of Fall armyworm (FAW) Spodoptera frugiperda reared in the laboratory with three monocot plant diets (Sugarcane [M1], maize [M2] and onion [M3]) using 16S rRNA gene sequencing. The study revealed variations in the structure and community of the gut bacterial biota of FAW larvae reared with three monocot plants. Diet sources greatly altered the structure of the gut bacterial community. Three alpha diversity indices (observed species richness Chao1 and ACE) showed significant differences among the various diet groups. The principal component analysis revealed a moderate distinction in bacterial communities of the FAW larvae reared with the three monocot plants. Higher numbers of operational taxonomic units (OTUs) were observed in sugarcane diet (M1). The bacterial communities of all the larvae groups were dominated by phylum Firmicutes. A redundancy analysis (RDA) indicated that neutral detergent fibre (NDF) (p < 0.0005) was the most significant factors influencing the community distribution of gut bacteria of FAW larvae. Functional Annotation of Prokaryotic Taxa (FAPROTAX) analysis predicted that chemoheterotrophy, fermentation, aerobic chemoheterotrophy and aromatic compound degradation were the prominent putative functions of FAW larvae gut bacteria communities. Chemoheterotrophy was the most dominant putative functions predicted and was higher in sugarcane diet. Furthermore, the complexity of the network structure was higher in M1 while the modularity was higher in maize diet. The different monocot diets had a considerable impact on bacterial composition in the gut of Fall armyworm larvae and their ecological and functional relevance are discussed.

Plants are equipped with various defensive attributes against herbivores, including volatile and nonvolatile compounds. In maize plants, benzoxazinoids mediate resistance against some herbivores, with the most abundant being (2R)-2-β-D-glucopyranosyloxy-4-hydroxy-7-methoxy-2H-1,4-benzoxazin3(4H)-one (DIMBOA-Glc), and its corresponding aglucone 2,4-dihydroxy-7-methoxy-2H-1,4-benzoxazin-3(4H)-one (DIMBOA). Both compounds have been shown to interfere in the larval development of generalist herbivores but are less effective on specialist, that is, grass-feeding, herbivores. Using a Brazilian population of Spodoptera frugiperda, we investigated (i) the level of constitutive benzoxazinoids in Neotropical maize genotypes, that is, Zapalote Chico, Mirt 2A, Sintético Spodoptera, L3, BRS 4103 and BRS 1040 (ii) the effect of S. frugiperda herbivory on benzoxazinoid levels in these genotypes and (iii) the impact of the genotypes on the development of S. frugiperda larvae. The results showed that the six maize genotypes produce different levels of benzoxazinoids, with Mirt 2A and BRS 1040 producing constitutively higher levels of HDMBOA-Glc and DIMBOA-Glc respectively compared to the other genotypes. When feeding on BRS 1040 and Mirt 2A, S. frugiperda larvae took an additional week to pupate, but this effect does not affect larval survival, what was the same and high on all the genotypes (>70%). Furthermore, production of DIMBOA-Glc and HDMBOA-Glc in these genotypes was suppressed, suggesting that S. frugiperda larvae can alter maize defence plant responses. In summary, our results demonstrate that Neotropical maize genotypes produce varying levels of benzoxazinoids, genotypes respond differently to S.frugiperda herbivory and S. frugiperda is able to cope with secondary metabolite-based defence in Neotropical maize. Made available in DSpace on 2022-05-16T13:13:18Z (GMT). No. of bitstreams: 1 Physiological-Entomology-2022-Michereff-Neotropical-maize-genotypes-with-different-levels-of-benzoxazinoids-affect.pdf: 962114 bytes, checksum: cbbcdb1f0711b41ceaa6476cd8972046 (MD5) Previous issue date: 2022 Na publicação - Maria Carolina Blassioli-Moraes.

Crop damage caused by fall armyworm (FAW), Spodoptera frugipera J.E. Smith (1797) (Lepidoptera: Noctuidae), has generated concern among agriculturists globally. In 2019, FAW was first reported in Sri Lanka, where it caused significant losses to corn crops. However, given that the two FAW biotypes – “rice strain” (R-FAW) and “corn strain” (C-FAW) – are morphologically identical, the biotype(s) present in Sri Lanka were unknown. The current study used the mitochondrial cytochrome c oxidase subunit I gene (mt-CO1) of FAW to biotype nine FAW samples collected in Sri Lanka. The resulting molecular phylogeny revealed that both R-FAW and C-FAW biotypes were present among the samples. In addition, we used the temperature-switch polymerase chain reaction (PCR) technique to develop a gel-based molecular marker. Two fragments were successfully amplified by the newly developed marker ABUOP0002, with fragment sizes of 341 bp from R-FAW and 204 bp from C-FAW. This demonstrates that ABUOP0002 can serve as a diagnostic gel-based molecular marker to identify the R-FAW and C-FAW biotypes from samples already identified to species level as S. frugiperda through taxonomical keys and provides a possible alternative to more expensive sequencing-based assays.

Insects manifest phenotypic plasticity in their development and behavior in response to plant defenses, via molecular mechanisms that produce tissue-specific changes. Phenotypic changes might vary between species that differ in their preferred hosts and these effects could extend beyond larval stages. To test this, we manipulated the diet of southern armyworm (SAW; Spodoptera eridania) and fall armyworm (FAW; Spodoptera frugiperda) using a tomato mutant for jasmonic acid plant defense pathway (def1), and wild type plants, and then quantified gene expression of Troponin t (Tnt) and flight muscle metabolism of the adult insects. Differences in Tnt spliceform ratios in insect flight muscles correlate with changes to flight muscle metabolism and flight muscle output. We found that SAW adults reared on induced def1 plants had higher relative abundances (RA) of the A isoform of Troponin t (Tnt A) in their flight muscles; in contrast, FAW adults reared on induced def1 plants had lower RAs of Tnt A in their flight muscles compared to adults reared on def1 and controls. Although mass-adjusted flight metabolic rates showed no independent host plant effects in either species, higher flight metabolic rates in SAW correlated with increased RAs of Tnt A. Flight muscle metabolism also showed an interaction of host plants with Tnt A in both species, suggesting that host plants might be influencing flight muscle metabolic output by altering Tnt. This study illustrates how insects respond to variation in host plant's chemical defense by phenotypic modifications to their flight muscle proteins, with possible implications for dispersal.

RNA interference (RNAi) is a promising technology for the development of next-generation insect pest control products. Though RNAi is efficient and systemic in coleopteran insects, it is inefficient and variable in lepidopteron insects. In this study, we explored the possibility of improving RNAi in the fall armyworm (FAW), Spodoptera frugiperda by conjugating double-stranded RNA (dsRNA) with biodegradable chitosan (Chi). dsRNA conjugated with chitosan was protected from degradation by endonucleases present in Sf9 cell-conditioned medium, hemolymph, and midgut lumen contents collected from the FAW larvae. Chi-dsRNA complexes showed reduced accumulation in the endosomes of Sf9 cells and FAW tissues. Exposing chitosan formulated dsRNA in Sf9 cells and the tissues induced a significant knockdown of endogenous genes. Chi-dsIAP fed to FAW larvae induced knockdown of iap gene, growth retardation, and mortality. Processing of dsRNA into small interfering RNA was detected with chitosan-conjugated 32 P-UTP-labeled ds green fluorescent protein in Sf9 cells and FAW larval tissues. Overall, these data suggest that dsRNA conjugated with chitosan helps dsRNA escape from the endosomes and improves RNAi efficiency in FAW cells and tissues.

Fall armyworm (FAW; Spodoptera frugiperda), an exotic moth which recently invaded Africa, is a highly destructive pest of cereals especially maize a highly valued staple crop in Nigeria. The use of natural enemies such as predators or parasitoids for FAW control is more economically viable and environmentally safer than currently recommended synthetic insecticides. Natural enemies to combat the pest have not yet been reported in Nigeria. An exploration for the pests’ natural enemies was undertaken by collecting FAW eggs and larvae from maize fields. These were reared in the laboratory for emergence, identification and efficacy as natural enemies. This yielded Euplectrus laphygmae (Hymenoptera: Eulophidae); Telenomus remus (Hymenoptera: Platygastridae) and Trombidium sp. (Acari.: Trombidiidae). Cotesia or Apanteles spp. were inferred to occur since Stictopisthus sp. (Hym.: Ichneumonidae), a secondary parasitoid, that attacks cocoons of Microgasterinae (e.g. Cotesia, Apanteles etc.) also emerged. Species of yet-to-be identified predators were also observed in various niches of maize plants. A positive relationship was found between FAW instar and the number of E. laphygmae eggs/instar ranging, on average, from 1.5 on second instar to 5.5 on fourth instars hosts. Parasitism rate of T. remus on FAW eggs was 100%. Parasitic mite infestation resulted in increasing paleness, reduced feeding, growth and movement as well as death of FAW 1st instars. Thus, the occurrence of FAW natural enemies in Nigeria calls for advocacy campaign to incorporate their use into integrated pest management strategies that attract and allow natural enemies to thrive for FAW management.

Mechanical damage caused by insect feeding along with components present in insect saliva and oral secretions are known to induce jasmonic acid–mediated defense responses in plants. This study investigated the effects of bacteria from oral secretions of the fall armyworm Spodoptera frugiperda on herbivore-induced defenses in tomato and maize plants. Using culture-dependent methods, we identified seven different bacterial isolates belonging to the family Enterobacteriacea from the oral secretions of field-collected caterpillars. Two isolates, Pantoea ananatis and Enterobacteriaceae-1, downregulated the activity of the plant defensive proteins polyphenol oxidase and trypsin proteinase inhibitors (trypsin PI) but upregulated peroxidase (POX) activity in tomato. A Raoultella sp. and a Klebsiella sp. downregulated POX but upregulated trypsin PI in this plant species. Conversely, all of these bacterial isolates upregulated the expression of the herbivore-induced maize proteinase inhibitor (mpi) gene in maize. Plant treatment with P. ananatis and Enterobacteriaceae-1 enhanced caterpillar growth on tomato but diminished their growth on maize plants. Our results highlight the importance of herbivore-associated microbes and their ability to mediate insect plant interactions differently in host plants fed on by the same herbivore.

To grow and thrive plants must be able to adapt to both adverse environmental conditions and attack by a variety of pests. Elucidating the sophisticated mechanisms plants have developed to achieve this has been the focus of many studies. What is less well understood is how plants respond when faced with multiple stressors simultaneously. In this study, we assess the response of Zea mays (maize) to the combinatorial stress of flooding and infestation with the insect pest Spodoptera frugiperda (fall armyworm). This combined stress leads to elevated production of the defence hormone salicylic acid, which does not occur in the individual stresses, and the resultant salicylic acid-dependent increase in S. frugiperda resistance. Remodelling of phenylpropanoid pathways also occurs in response to this combinatorial stress leading to increased production of the anti-insect C-glycosyl flavones (maysins) and the herbivore-induced volatile phenolics, benzyl acetate, and phenethyl acetate. Furthermore, changes in cellular redox status also occur, as indicated by reductions in peroxidase and polyphenol oxidase activity. These data suggest that metabolite changes important for flooding tolerance and anti-insect defence may act both additively and synergistically to provide extra protection to the plant.

The perception of herbivory by plants is known to be triggered by the deposition of insect-derived factors such as saliva and oral secretions, oviposition materials, and even feces. Such insect-derived materials harbor chemical cues that may elicit herbivore and/or pathogen-induced defenses in plants. Several insect-derived molecules that trigger herbivore-induced defenses in plants are known; however, insect-derived molecules suppressing them are largely unknown. In this study, we identified two plant chitinases from fall armyworm (Spodoptera frugiperda) larval frass that suppress herbivore defenses while simultaneously inducing pathogen defenses in maize (Zea mays). Fall armyworm larvae feed in enclosed whorls of maize plants, where frass accumulates over extended periods of time in close proximity to damaged leaf tissue. Our study shows that maize chitinases, Pr4 and Endochitinase A, are induced during herbivory and subsequently deposited on the host with the feces. These plant chitinases mediate the suppression of herbivore-induced defenses, thereby increasing the performance of the insect on the host. Pr4 and Endochitinase A also trigger the antagonistic pathogen defense pathway in maize and suppress fungal pathogen growth on maize leaves. Frass-induced suppression of herbivore defenses by deposition of the plant-derived chitinases Pr4 and Endochitinase A is a unique way an insect can co-opt the plant's defense proteins for its own benefit. It is also a phenomenon unlike the induction of herbivore defenses by insect oral secretions in most host-herbivore systems.

Insect immunity includes a surveillance system that detects and signals infections, coupled with hemocytic and humoral immune functions. These functions are signaled and coordinated by several biochemicals, including biogenic amines, insect cytokines, peptides, and prostaglandins (PGs). The actions of these mediators are coordinated within cells by various forms of cross-talk among the signaling systems and they result in effective reactions to infection. While this is well understood, we lack information on how immune-mediated recovery influences subsequent juvenile development in surviving insects. We investigated this point by posing the hypothesis that PG signaling is necessary for larval recovery, although the recovery imposes biological costs, registered in developmental delays and failures in surviving individuals. Here, we report that nodulation responses to infections by the bacterium, Serratia marcescens, increased over time up to 5 h postinfection, with no further nodulation; it increased in a linear manner with increasing bacterial dosages. Larval survivorship decreased with increasing bacterial doses. Treating larvae with the PG-biosynthesis inhibitor, indomethacin, led to sharply decreased nodulation reactions to infection, which were rescued in larvae cotreated with indomethacin and the PG-precursor, arachidonic acid. Although nodulation was fully rescued, all bacterial challenged larvae suffered reduced survivorship compared to controls. Bacterial infection led to reduced developmental rates in larvae, but not pupae. Adult emergence from pupae that developed from experimental larvae was also decreased. Taken together, our data potently bolster our hypothesis.

Selecting optimal host plants is critical for herbivorous insects, such as fall armyworm (Spodoptera frugiperda), an important maize pest in the Americas and Africa. Fall armyworm larvae are presumed to have limited mobility, hence female moths are presumed to be largely responsible for selecting hosts. We addressed host selection by fall armyworm moths and neonate and older (3rd-instar) larvae, as mediated by resistance and herbivory in maize plants. Thus, we compared discrimination among three maize cultivars with varying degrees of resistance to fall armyworm, and between plants subjected or not to two types of herbivory. The cultivars were: (i) susceptible, and deficient in jasmonic acid (JA) production and green leaf volatiles (GLV) emissions (inbred line B73-lox10); (ii) modestly resistant (B73), and; (iii) highly resistant (Mp708). The herbivory types were: (i) ongoing (= fall armyworm larvae present), and; (ii) future (= fall armyworm eggs present). In choice tests, moths laid more eggs on the highly resistant cultivar, and least on the susceptible cultivar, though on those cultivars larvae performed poorest and best, respectively. In the context of herbivory, moths laid more eggs: (i) on plants subject to versus free of future herbivory, regardless of whether plants were deficient or not in JA and GLV production; (ii) on plants subject versus free of ongoing herbivory, and; (iii) on plants not deficient in compared to deficient in JA and GLV production. Neonate larvae dispersed aerially from host plants (i.e. ballooned), and most larvae colonized the modestly resistant cultivar, and fewest the highly resistant cultivar, suggesting quasi-directional, directed aerial descent. Finally, dispersing older larvae did not discriminate among the three maize cultivars, nor between maize plants and (plastic) model maize plants, suggesting random, visually-oriented dispersal. Our results were used to assemble a model of host selection by fall armyworm moths and larvae, including recommendations for future research.

The underlying adaptive mechanisms by which insect strains are associated with specific plants are largely unknown. In this study, we investigated the role of herbivore-induced defenses in the host plant association of fall armyworm (Spodoptera frugiperda) strains. We tested the expression of herbivore-induced defense-related genes and the activity of plant-defensive proteins in maize and Bermuda grass upon feeding by fall armyworm strains. The rice strain caterpillars induced greater accumulation of proteinase inhibitors in maize than the corn strain caterpillars. In Bermuda grass, feeding by the corn strain suppressed induction of trypsin inhibitor activity whereas the rice strain induced greater activity levels. Differences in elicitation of these plant defenses by the two strains seems to be due to differences in the activity levels of the salivary enzyme phospholipase C. The levels of plant defense responses were negatively correlated with caterpillar growth, indicating a fitness effect. Our results indicate that specific elicitors in the saliva of fall armyworm stains trigger differential levels of plant defense responses that affect caterpillar growth and thus may influence host plant associations in field conditions. The composition and secretion of plant defense elicitors may have a strong influence in the host plant association of insect herbivores.

The effect of the insecticide endosulfan on the predatory efficiency of the green lacewing, Chrysoperla carnea Stephens (Neuroptera: Chrysopidae) on the eggs of tobacco budworm, Heliothis virescens (Fabricius) (Lepidoptera: Noctuidae) and fall armyworm, Spodoptera frugiperda (Smith) (Lepidoptera: Noctuidae) was measured with Holling’s disc equation. Though the type II functional response was maintained in C. carnea exposed to endosulfan, the functional response parameters: attack rate (a′), handling time (Th), total handling time (Tht), searching time (Ts), and search efficiency (E) were affected for both prey offered. The predator took more time to identify, pursue, capture, consume, and digest the prey, and in general, the efficiency of the predator as a biological control agent was adversely affected.

The interference imposed the by weeds on corn decreases practically all vegetative characteristics. As consequence, the green ear and grain yield are also reduced. Losses due to the fall armyworm (Spodoptera frugiperda) attack can reduce corn grain yield up to 34%. In general, weed and insect control issues are addressed separately in research papers. Nevertheless, interaction between weeds and insects may exist. This study aimed to evaluate green ear and corn grain yield response to weed and fall armyworm control. A completely randomized block design with split-plots and five replicates was adopted. Corn cultivar AG 1051 was grown under weedy conditions or with control by hand hoeings performed at 20 and 40 days after planting. Fall armyworm control (applied to subplots) was performed with sprays of water (control), deltamethrin (5g active ingredient ha-1); neem oil, at 0.5% (diluted in water), and neem leaf extract at 5%. Each product was sprayed three times, at seven-day intervals, starting at the 7th day after planting, using 150 L ha-1 of the tank solution. Dry mass of the above-ground part, internode diameter, leaf length, leaf width, leaf area, green ear yield and grain yield of corn were reduced due to the lack of weed control. Fall armyworm control in the weeded plots did not influence green ear yield and grain yield, except green mass of marketable, husked ears, which was reduced when the caterpillar was not controlled. Without weed control, neem extracts and deltamethrin sprays provided highest yields of number and total weight of green ears with husks, number and weight of marketable ears with husks and number of marketable ears without husks. The best results for husked ear mass and for grain yield were obtained with neem extract and deltamethrin, respectively.

The signaling pathways that enable plants to mount defenses against insect herbivores are known to be complex. It was previously demonstrated that the insect-resistant maize (Zea mays L.) genotype Mp708 accumulates a unique defense cysteine proteinase, Mir1-CP, in response to caterpillar feeding. In this study, the role of ethylene in insect defense in Mp708 and an insect-susceptible line Tx601 was explored. Ethylene synthesis was blocked with either cobalt chloride or aminoethoxyvinylglycine. Alternatively, ethylene perception was inhibited with 1-methylcyclopropene. Blocking ethylene synthesis and perception resulted in Mp708 plants that were more susceptible to caterpillar feeding. In addition, fall armyworm (Spodoptera frugiperda) larvae that fed on inhibitor-treated Mp708 plants had signifycantly higher growth rates than those reared on untreated plants. In contrast, these responses were not significantly altered in Tx601. The ethylene synthesis and perception inhibitors also reduced the accumulation of Mir1-CP and its transcript mir1 in response to herbivory. These results indicate that ethylene is a component of the signal transduction pathway leading to defense against insect herbivory in the resistant genotype Mp708.

Department of Entomology, 333 Leon Johnson Hall, Montana State University, Bozeman, Montana 59717, United States of America Macedo et al. 500 The wheat stem sawfly, Cephus cinctus Norton (Hymenoptera: Cephidae), is an insect pest in dryland wheat cropping systems in the southern Canadian Prairies and the northern Great Plains of the United States (Morrill 1997). Yield losses caused by C. cinctus are due to reduced head weight (Holmes 1977; Morrill et al. 1992) and lodging, which decreases harvest efficiency. Estimates of yield losses in Montana alone are about US$25 million per year. The biology of C. cinctus is well known. It has a wide host range including nearly all large-stemmed grasses except oats, Avena sativa L. (Farstad 1940). Adults emerge from fallow fields and migrate into crop fields as wheat stems elongate, from mid-May to late July (Morrill and Kushnak 1999). Eggs are laid in stems, and larvae feed on the parenchyma (Ainslie 1920) of growing wheat plants. Plants are susceptible from stem elongation through flowering. Mature larvae move to the base of the plant and cut notches around the inner perimeter of the stem near ground level, which usually causes lodging and loss of wheat heads at harvest (Ainslie 1920). The larvae overwinter in diapause within the lower section of the cut stem (Ainslie 1920). Diapause is completed after about 3 months at low temperature (Salt 1947), and pupation occurs in the spring. New basic research studies would be possible if C. cinctus larvae could be reared on artificial media. Currently, larvae can be reared only in plant stems, which must be reared in a glasshouse or be collected from the field. Both topical and consumption dose-response studies are difficult to quantify with insects that are feeding within plant stems. Off-season adults can be obtained by refrigerating field-collected diapausing larvae. Pupation and adult eclosion will occur following incubation at room temperature. Adults readily mate in the laboratory, and artificial oviposition sites can be provided to obtain eggs. The first reported attempt to rear C. cinctus larvae on artificial diet used natural foods such as wheat juice and cellulose powder (Kasting and McGinnis 1958). Although C. cinctus were reared on a diet mixture (Kasting and McGinnis 1958), larval survival was limited to 14 days, and it was necessary to provide fresh diet every 24–48 h (McGinnis and Kasting 1962). Identified problems included mold contamination and the configuration of the diet (McGinnis and Kasting 1962). The legless C. cinctus larvae are adapted to feeding within an enclosed tube, or plant stem. To simulate these conditions, McGinnis and Kasting (1962) improved larval survival by placing evenly spaced plastic plates with grooves in the media. Since then, there have been no reports of improved methods for rearing C. cinctus larvae. Development of a satisfactory artificial medium, based on a better understanding of nutritional requirements and presentation of diet, would open new opportunities for our current research on larval antibiosis and fungal pathogens. This paper presents a new diet medium for the wheat stem sawfly that was successfully used to rear neonate larvae to maturation. In the first experiment, a commercial diet for fall armyworm, Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae), consisting of sucrose, cholesterol, methylparaben, sorbic acid, vitamin mix (Vanderzant), ascorbic acid, stabilized wheat germ, casein, sterile corn cob grits, Wesson salt mix, and raw linseed oil (diet F9179B, BioServ, Frenchtown, New Jersey), was tested. The dry mixture was combined with 1 L of boiling agar (A-7002, SIGMA Chemical Co., St. Louis, Missouri) and blended for 2 min. The final mixture was dispensed immediately into 60 mm × 15 mm sterilized disposable plastic petri dishes (Fisher brand, Fisher

The circadian rhythms of locomotor activity were studied in larvae and adult Fall Armyworms, Spodoptera frugiperda, fed a nutritional diet (control) or a diet containing the maize variety CML67 (experimental). Activity was monitored using infrared-light crossings and rhythms were evaluated using actograms, average waveform, and X2 periodograms. Results show that larvae grown on the experimental diet did not display conspicuous circadian rhythms in constant darkness before pupation and showed poor responses to light-dark cycles. Significant differences between the two groups were observed in their maximal activity and activity-rest ratios. Adults fed either diet displayed circadian rhythms of locomotor activity, however, differences were still found in their activity-rest ratios. Results obtained indicate that animals fed with diet containing the maize variety CLM67, have significant differences in the expression of circadian locomotor activity in larvae under constant darkness and in their responses to ar...

In addition to feeding damage, herbivores release cues that are recognized by plants to elicit defenses. Caterpillar oral secretions have been shown to trigger herbivore defense responses in several different plant species. In this study, the effects of two fall armyworm (Spodoptera frugiperda) oral secretions (saliva and regurgitant) on caterpillar defense responses in maize (Zea mays) were examined. Only minute amounts of regurgitant were deposited on the maize leaf during larval feeding bouts and its application to leaves failed to induce the expression of several herbivore defense genes. On the other hand, caterpillars consistently deposited saliva on leaves during feeding and the expression of several maize defense genes significantly increased in response to saliva application and larval feeding. However, feeding by ablated caterpillars with impaired salivation did not induce these defenses. Furthermore, bioassays indicated that feeding by unablated caterpillars significantly enhanced defenses when compared with that of ablated caterpillars. Another critical finding was that the maize genotype and stage of development affected the expression of defense genes in response to wounding and regurgitant treatments. These results demonstrate that fall armyworm saliva contains elicitors that trigger herbivore defenses in maize.

Summary Some plant-derived anti-herbivore defensive proteins are induced by insect feeding, resist digestion in the caterpillar gut and are eliminated in the frass. We have identified several maize proteins in fall armyworm (Spodoptera frugiperda) frass that potentially play a role in herbivore defense. Furthermore, the toxicity of one of these proteins, ribosome-inactivating protein 2 (RIP2), was assessed and factors regulating its accumulation were determined. To understand factors regulating RIP2 protein accumulation, maize (Zea mays) plants were infested with fall armyworm larvae or treated with exogenous hormones. The toxicity of recombinant RIP2 protein against fall armyworm was tested. The results show that RIP2 protein is synthesized as an inactive proenzyme that can be processed in the caterpillar gut. Also, caterpillar feeding, but not mechanical wounding, induced foliar RIP2 protein accumulation. Quantitative real-time PCR indicated that RIP2 transcripts were rapidly induced (1 h) and immunoblot analysis indicated that RIP2 protein accumulated soon after attack and was present in the leaf for up to 4 d after caterpillar removal. Several phytohormones, including methyl jasmonate, ethylene, and abscisic acid, regulated RIP2 protein expression. Furthermore, bioassays of purified recombinant RIP2 protein against fall armyworm significantly retarded caterpillar growth. We conclude that the toxic protein RIP2 is induced by caterpillar feeding and is one of a potential suite of proteins that defend maize against chewing herbivores.

Plants respond to insect herbivory through the production of biochemicals that function as either direct defenses or indirect defenses via the attraction of natural enemies. While attack by closely related insect pests can result in distinctive levels of induced plant defenses, precise biochemical mechanisms responsible for differing responses remain largely unknown. Cowpea (Vigna unguiculata) responds to Fall armyworm (Spodoptera frugiperda) herbivory through the detection of fragments of chloroplastic ATP synthase γ-subunit proteins, termed inceptin-related peptides, present in larval oral secretions (OS). In contrast to generalists like Fall armyworm, OS of the legume-specializing velvetbean caterpillar (VBC; Anticarsia gemmatalis) do not elicit ethylene production and demonstrate significantly lower induced volatile emission in direct herbivory comparisons. Unlike all other Lepidoptera OS examined, which preferentially contain inceptin (Vu-In; +ICDINGVCVDA−), VBC OS contain predominantly a C-terminal truncated peptide, Vu-In−A (+ICDINGVCVD−). Vu-In−A is both inactive and functions as a potent naturally occurring antagonist of Vu-In-induced responses. To block antagonist production, amino acid substitutions at the C terminus were screened for differences in VBC gut proteolysis. A valine-substituted peptide (Vu-InƊV; +ICDINGVCVDV−) retaining full elicitor activity was found to accumulate in VBC OS. Compared with the native polypeptide, VBC that previously ingested 500 pmol of the valine-modified chloroplastic ATP synthase γ-subunit precursor elicited significantly stronger plant responses in herbivory assays. We demonstrate that a specialist herbivore minimizes the activation of defenses by converting an elicitor into an antagonist effector and identify an amino acid substitution that recovers these induced plant defenses to a level observed with generalist herbivores.

The influence of dietary allelochemical on ecdysone 20-monooxygenase activity was studied in the fall armyworm, Spodoptera frugiperda (J.E. Smith). Feeding the indoles (indole-3-carbinol, indole-3-acetonitrile), flavonoids (flavone, β-naphthoflavone), monoterpenes (menthol, menthone, peppermint oil), and a coumarin (xanthotoxin) to the larvae stimulated midgut microsomal ecdysone 20-monooxygenase activity from 28 to 200% as compared with the controls. β-Naphthoflavone was the most potent inducer among those tested. Phenobarbital, a well-known cytochrome P450 inducer, also caused a 2-fold increase in the microsomal ecdysone 20-monooxygenase activity. Ecdysone 20-monooxygenase activity was 2.7-fold higher in the microsomal fraction than in the mitochondrial fraction isolated from larval midguts. Microsomal ecdysone 20-monooxygenase activity was highest in the fat body, followed by the midgut and Malpighian tubules. Tissue localization and enzyme inducibility were different between ecdysone 20-monooxygenase and xenobiotic-metabolizing cytochrome P450 monooxygenases, including aldrin epoxidase, biphenyl hydroxylase, methoxyresorufin O-demethylase, 7-ethoxycoumarin O-deethylase, p-chloro-N-methylaniline N-demethylase, and phorate sulfoxidase in fall armyworm larvae. © 1995 Wiley-Liss, Inc.

Five insecticide synergists, all of which were either methylenedioxyphenyl compounds or analogs, were compared as to their effect on cytochrome P450 monooxygenase induction caused by an allelochemical in fall armyworm larvae. Feeding the synergists (piperonyl butoxide, safrole, isosafrole, MGK 264, and myristicin) individually to the larvae caused decreases in the microsomal aldrin epoxidase activities ranging from 38% to 74% when compared with controls. Feeding indole-3-carbinol resulted in a 4-fold increase in the microsomal epoxidase activity. However, cotreatment of any of the synergists and the inducer completely eliminated the induction. Sixth instar larvae were more inducible than second instar larvae with respect to microsomal epoxidase and glutathione transferase in the fall armyworm. Enzyme inducibility varied widely among the seven phytophagous Lepidoptera examined. When indole-3-carbinol was used as an inducer of microsomal epoxidase, the extent of inducibility of the enzyme was fall armyworm > velvetbean caterpillar > corn earworm > beet armyworm > tobacco budworm > cabbage looper > diamondback moth. When indole-3-acetonitrile was used as an inducer, the inducibility of glutathione transferase was fall armyworm > beet armyworm > corn earworm > cabbage looper > velvetbean caterpillar > tobacco budworm > diamondback moth. Inducibility of five microsomal oxidase systems also varied considerably in the corn earworm, indicating the multiplicity of cytochrome P450 in this species. Microsomal epoxidase and glutathione transferase were induced by cruciferous host plants such as cabbage and their allelochemicals in diamondback moth larve. © 1993 Wiley-Liss, Inc.

Mitochondrial DNA from the fall armyworm, Spodoptera frugiperda (J.E. Smith), was cloned and characterized using restriction enzyme mapping. Genome size is approximately 16.3 kilobase (Kb), consistent with that of most animals. Three fragments, 8.1 Kb, 6.2 Kb, and 2.0 Kb, were produced by digestion with restriction enzyme Xbal and cloned into a pUC19 vector. Twenty-nine restriction enzymes were used to generate a detailed physical restriction enzyme map of the three cloned fragments. These data represent the first detailed characterization of a lepidopteran mitochondrial genome. © 1992 Wiley-Liss, Inc.

In plants, ethylene and jasmonate control the defense responses to multiple stressors, including insect predation. Among the defense proteins known to be regulated by ethylene is maize insect resistance 1-cysteine protease (Mir1-CP). This protein is constitutively expressed in the insect-resistant maize (Zea mays) genotype Mp708; however, its abundance significantly increases during fall armyworm (Spodoptera frugiperda) herbivory. Within 1 h of herbivory by fall armyworm, Mir1-CP accumulates at the feeding site and continues to increase in abundance until 24 h without any increase in its transcript (mir1) levels. To resolve this discrepancy and elucidate the role of ethylene and jasmonate in the signaling of Mir1-CP expression, the effects of phytohormone biosynthesis and perception inhibitors on Mir1-CP expression were tested. Immunoblot analysis of Mir1-CP accumulation and quantitative reverse-transcriptase polymerase chain reaction examination of mir1 levels in these treated plants demonstrate that Mir1-CP accumulation is regulated by both transcript abundance and protein expression levels. The results also suggest that jasmonate functions upstream of ethylene in the Mir1-CP expression pathway, allowing for both low-level constitutive expression and a two-stage defensive response, an immediate response involving Mir1-CP accumulation and a delayed response inducing mir1 transcript expression.

Caterpillars of the noctuid moth, Spodoptera frugiperda (J. E. Smith), reared on artificial diets diluted with cellulose and water, increased fresh weight (fw) consumption 2.2–2.5-fold over those on the undiluted diet. At moderate levels of water- or cellulose-dilution, the increased consumption, combined with increased digestion and absorption of nutrients (ADNU), sufficiently compensated for the reduced nutrient intake to achieve pupal biomass equivalent to that of pupae from the undiluted diet. At higher levels of water- and cellulose-dilution, fw consumption and ADNU increased further but pupal dry weight declined on the water-diluted diets. At each level of dilution fw consumption and ADNU increased similarly on the water- and cellulose-diluted diets, but biomass gain was reduced on the water- compared with the cellulose-diluted diets. This was due in part to lowered food conversion efficiency on the water-diluted diets, possibly caused by increased costs of metabolizing the wetter diets. Our data support the hypothesis that consumption rates are regulated by a nutrient feedback mechanism. Our data also suggest that digestive enzyme activity is correlated with consumption. Furthermore, the cost of increased consumption rates on diets of reduced energetic value may constitute a substantially greater energy expenditure than previously believed. However, this cost was insufficient to reduce relative growth rates but was apparently manifested in lowered lipid accumulation.

Submitted by (edna.saturno@ufrpe.br) on 2016-11-23T13:13:53Z No. of bitstreams: 1 Alicely Araujo Correia.pdf: 1178729 bytes, checksum: 007e1cb0c61d3a8f7fdcf0344b043ecf (MD5) Made available in DSpace on 2016-11-23T13:13:53Z (GMT). No. of bitstreams: 1 Alicely Araujo Correia.pdf: 1178729 bytes, checksum: 007e1cb0c61d3a8f7fdcf0344b043ecf (MD5) Previous issue date: 2008-02-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) is the main pest of corn crops. The control of S. frugiperda requires various insecticide applications that might result on selection of resistant populations and environmental contamination. A promising alternative toward the control of this pest is the use of vegetal insecticides such as neem, Azadirachta indica A. Juss (Meliaceae). Few data exist related to the immunologic and histophysiologic reactions of S. frugiperda larvae under neem treatment. Thus, the present research had as objectives: (a) to characterize and to quantify the hemocytes of S. frugiperda larvae pre- and post-treatment with concentrations 0.5 and 1.0% of neem (Neemseto®) on the intervals of 48, 96, 144, 192 and 240h; (b) to describe, histologically, the food channel of the larvae; (c) to characterize the effect of the concentrations 0.5 and 1.0% of neem, in the same evaluation intervals after treatment, on the histophysiology of midgut; (d) to quantify the regenerative cells and; (e) to analyze,histochemically, the secretion product of midgut. Six types of hemocytes were identified in the hemolymph of S. frugiperda larvae: Adipohemocytes, Spherulocytes, Granulocytes, Eonocytoids, Plasmatocytes and Prohemocytes. And, the most abundant hemocytes were Granulocytes and Plasmatocytes. The counting of hemocytes showed effects as function of neem concentrations onthe hemocytary dynamics and possivelmente the endocrine events, contributing for mortality and observed morphologic abnormalities. Despite of expressive larval mortality and of effects with neem at concentration 1.0%, both concentrations (0.5 and 1.0%) modified the cellular immunity of the larvae. Histophysiological alterations occurred as function of tested neem concentrations and time after treatment observed as: degeneration of the epithelium and musculature of the midgut, reduction of regenerative cells and the secretory activity of this region, in the two concentrations of neem. These alterations, however, were observed for larvae treated with neem at concentration 1.0% already from 96h after treatment. These results demonstrate the insecticide activity of the commercial formulation neem (Neemseto®) in both tested concentrations, on thehistophysiology of midgut and on the immunologic reactions of S. frugiperda larvae. Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) é a principal praga da cultura do milho. Seu controle demanda elevado número de aplicações de inseticidas sintéticos, podendo ocasionar seleção de populações resistentes e contaminação ambiental. Uma alternativa promissora para o controle deste inseto é a utilização de substâncias com ação inseticida oriundas de plantas como o nim, Azadirachta indica A. Juss (Meliaceae). Contudo, os aspectos relacionados às reações imunológicas e histofisiológicas de lagartas de S. frugiperda, promovidos pela interação com o nim, ainda são pouco estudados. Assim, a presente pesquisa teve como objetivos: (a) caracterizar e realizar a contagem diferencial dos hemócitos de lagartas de S. frugiperda, antes e após o tratamento com nim (Neemseto®) nas concentrações 0,5 e 1,0%, nos intervalos de 48, 96, 144, 192 e 240h; (b )descrever, histologicamente, o canal alimentar dessas lagartas; (c) caracterizar o efeito das concentrações 0,5 e 1,0% do nim, nos mesmos intervalos deavaliação após tratamento, sobre a histofisiologia do mesêntero; (d) quantificar as células regenerativas e; (e) analisar, histoquimicamente, o produto de secreção do mesêntero. Na hemolinfa das lagartas de S. frugiperda foram identificados seis tipos de hemócitos: Adipohemócitos, Esferulócitos, Granulócitos, Oenocitóides, Plasmatócitos e Prohemócitos, sendoos Granulócitos e Plasmatócitos mais freqüentes. A contagem diferencial dos hemócitos revelou um efeito concentração-dependente do nim, afetando a dinâmica hemocitária e possivelmente os eventos endócrinos, contribuindo para a mortalidade e anormalidades morfológicas observadas. Apesar da mortalidade e dos efeitos ocorrerem de forma mais expressiva no tratamento com nim a 1,0%, ambas as concentrações (0,5 e 1,0%) alteraram a imunidade celular das lagartas. Foram observadas alterações histofisiológicas que variaram de intensidade de acordo com o tempo e as concentrações, tais como: degeneração do epitélio e da musculatura do mesêntero, redução de células regenerativas e da atividade secretora desta região, nas duas concentrações do nim. Porém, estas alterações foram observadas nas lagartas tratadas com nim a 1,0% já a partir de 96h após o tratamento. Esses resultados demonstram a atividade inseticida da formulação comercial de nim(Neemseto) em ambas as concentrações testadas, na histofisiologia do mesêntero e nas reações imunológicas de lagartas de S. frugiperda.

Plants have multiple strategies to deal with herbivory, ranging from chemical or physical defenses to tolerating damage and allocating resources for regrowth. Grasses usually tolerate herbivory, but for some cool-season grasses, their strategy may depend upon their interactions with intracellular symbionts. Neotyphodium endophytes are common symbionts in pooid grasses, and, for some host species, they provide chemical defenses against both vertebrate and invertebrate herbivores. Here, it was tested whether defenses provided by Neotyphodium coenophialum in Lolium arundinaceum (tall fescue) are inducible by both mechanical damage and herbivory from an invertebrate herbivore, Spodoptera frugiperda (fall armyworm), via a bioassay and by quantifying mRNA expression for lolC, a gene required for loline biosysnthesis. Both mechanical and herbivore damage had a negative effect on the reproduction of a subsequent herbivore, Rhopalosiphum padi (bird cherry-oat aphid), and herbivore damage caused an up-regulation of lolC. Uninfected grass hosts also had significantly higher foliar N% and lower C:N ratio compared with infected hosts, suggesting greater allocation to growth rather than defense. For L. arundinaceum, N. coenophialum appears to switch its host's defensive strategy from tolerance via compensation to resistance.

O objetivo deste trabalho foi avaliar o efeito da mistura de herbicidas nicosulfuron + atrazine e o inseticida chlorpirifos sobre o cultivar de milho-pipoca UFVM2, as plantas daninhas e a lagarta-do-cartucho. Utilizou-se o delineamento experimental de blocos completos ao acaso, em esquema fatorial (5x2)+3, com quatro repetições. O primeiro fator foi constituído pelas doses de nicosulfuron (0, 10, 20, 30 e 40 g ha-1) + atrazine (1.200 g ha-1) + óleo mineral (900 g ha-1) e o segundo pelas doses do inseticida chlorpirifos (0 e 240 g ha-1). Foram avaliados três tratamentos adicionais: duas testemunhas, com e sem capina, ambas sem inseticida, e uma testemunha com capina e com inseticida. Não se observou interação significativa entre doses de nicosulfuron e do chlorpirifos no controle da lagarta-do-cartucho. A mistura no tanque formada pelo chlorpirifos com os herbicidas promoveu intoxicação às plantas de milho-pipoca, principalmente nas maiores doses do nicosulfuron (20, 30 e 40 g ha-1). O chlorpirifos não interferiu na eficiência de controle de plantas daninhas pelos herbicidas nem no estande final, no número de espigas por planta, no rendimento e na capacidade de expansão dos grãos de milho-pipoca. This study aimed to evaluate the effect of the tank-mixture herbicide nicosulfuron combined with the insecticide chlorpirifos on the popcorn cultivar UFVM2, weeds and fall armyworm. The experiment was arranged in a randomized complete block design, in a (5x2)+3 factorial scheme and four replications. The first factor consisted of nicosulfuron doses (0, 10, 20, 30, and 40 g ha-1) + atrazine (1.200 g ha-1) + oil (900 g ha-1) and the second of doses of the insecticide chlorpirifos (0 and 240 g ha-1). Three additional treatments were evaluated: two controls, with and without weeding, both without insecticide, and one control with weeding and insecticide. The interaction between the nicosulfuron and chlorpirifos doses in the control of the fall armyworm was not significant. The tank-mixture formed by the chlorpirifos and herbicides caused intoxication to popcorn plants, mainly at nicosulfuron rates (10, 20 and 40 g ha-1). Chlorpirifos did not interfere in herbicide efficiency in controlling weed, nor in the final stand, number of ears, and production and expansion capacity of the popcorn grains.