Roles of amino acids,protein, and fiber in leaf-feeding resistance of corn to the fall armyworm

The free amino acids have been shown by isolational work and choice bioassays to be more important than all other factors evaluated in defining leaf-feeding resistance of corn (Zea mays L.) to fall armyworm (FAW) [(Spodoptera frugiperda J.E. Smith)] larvae. 6-MBOA (6-methoxybenzoxazolinone) and maysin, toxins present in corn, were shown not to be significant factors for leaf-feeding resistance to first-instar FAW larvae because of their low concentrations in the whorl. Amino acid analysis showed that while the ratios of the essential amino acids in susceptible (S) and resistant (R) lines were similar, there were differences in the nonessential amino acids, particularly aspartic acid, which was higher in R lines. Also, the ratio of essential amino acids to nonessential amino acids was important, being too low in expressed whorl leaf juice (obtained from V₈–V₁₀ growth stage plants) to support larval growth, although juice was stimulatory in choice tests. The total protein content of whorls in S lines was about 15% higher than in R lines, but the significance of this difference is uncertain, because nutritional tests showed that larval growth increased with total protein only up to 12% protein. Sugars were only slightly stimulatory. Thus, the amino acids along with higher hemicellulose content of R lines, established by us earlier, appear to explain much of the basis of resistance in corn to larval leaf-feeding of the FAW.     

Hemicellulose is an important leaf-feeding resistance factor in corn to the fall armyworm

The fall armyworm,Spodoptera frugiperda (J. E. Smith), (FAW) is a major pest of corn,Zea mays L., in the southeastern United States. The damage to pretassel corn is caused by larvae feeding primarily on immature inner whorls. In this study, resistant lines were found to contain more crude fiber in whorls, mostly hemicellulose and cellulose. While hemicellulose, chiefly an arabinoxylan, was higher in resistant (R) lines than in susceptible (S) lines, the distribution of constituent neutral sugars was very similar in the lines. Both lines also containedp-coumaric and ferulic acids. These phenolic acids are known to occur both in the free state and in the cell wall as complexes bound by ester linkages to the arabinose moiety of the arabinoxylan.¹³C NMR data showed that the intensity of the carbonyl carbon (184 ppm) in resistant hemicellulose was stronger, indicating a greater degree of cross-linking. Thus, resistant hemicellulose is both structurally different from susceptible hemicellulose and present in greater quantities. In two of three laboratory dietary tests, FAW larval weight gains were significantly higher on diets with (S) hemicellulose incorporated at the same level as (R) hemicellulose. Therefore, resistance to the FAW appears to be correlated with both a greater amount and a higher degree of cross-linking of the hemicellulose of (R) lines.Mention of a trademark, proprietary product, or vendor does not constitute a guarantee of warranty of the product by the U.S. Department of Agriculture and does not imply its approval to the exclusion of other products or vendors that may also be suitable.     

A Maize Inbred Exhibits Resistance Against Western Corn Rootwoorm, <Emphasis Type="Italic">Diabrotica virgifera virgifera</Emphasis>

Insect resistance against root herbivores like the western corn rootworm (WCR, Diabrotica virgifera virgifera) is not well understood in non-transgenic maize. We studied the responses of two American maize inbreds, Mp708 and Tx601, to WCR infestation using biomechanical, molecular, biochemical analyses, and laser ablation tomography. Previous studies performed on several inbreds indicated that these two maize genotypes differed in resistance to pests including fall armyworm (Spodoptera frugiperda) and WCR. Our data confirmed that Mp708 shows resistance against WCR, and demonstrates that the resistance mechanism is based in a multi-trait phenotype that includes increased resistance to cutting in nodal roots, stable root growth during insect infestation, constitutive and induced expression of known herbivore-defense genes, including ribosomal inhibitor protein 2 (rip2), terpene synthase 23 (tps23) and maize insect resistance cysteine protease-1 (mir1), as well high constitutive levels of jasmonic acid and production of (E)-β-caryophyllene. In contrast, Tx601 is susceptible to WCR. These findings will facilitate the use of Mp708 as a model to explore the wide variety of mechanisms and traits involved in plant defense responses and resistance to herbivory by insects with several different feeding habits.     

Sources of fall armyworm,Spodoptera frugiperda (Lepidoptera: Noctuidae), kairomones eliciting host-finding behavior inCotesia (=Apanteles) marginivenitris (Hymenoptera: Braconidae)

Bioassay responses inCotesia marginiventris (Cresson) females to materials derived from fall armyworm (FAW) larvae,Spodoptera frugiperda (J. E. Smith), were most intense for frass and somewhat less intense for larval and pupal cutical materials, scales, exuviae, silk, and oral secretion, with FAW larval hemolymph eliciting only a slight response. The highest percentage of ovipositor probing was caused by frass (100%) and moth scales (90%). Various types of corn-leaf damage when assayed alone did not produce responses as intense as when assayed in combination with frass, cuticle material, and oral secretion. Parasitoid response was somewhat better to frass derived from FAW larvae feeding on corn and peanut leaves than from larvae feeding on the foliage of soybeans, Bermuda grass, cowpeas, or laboratory diet. Hexane and chloroform were better than methanol and water for extracting active material from FAW frass, and chloroform was the best of these solvents for extracting corn leaves. Serial dilutions of frass extracts resulted in a reduction in parasitoid response.This paper was submitted in partial fulfillment of the requirements for Doctor of Philosophy, Department of Entomology and Nematology, University of Florida, Gainesville.     

Behavioral and biological responses ofCotesia marginiventris to kairomones of the fall armyworm,Spodoptera frugiperda

Potential kairomone sources of the fall armyworm (FAW),Spodoptera frugiperda (J.E. Smith), were bioassayed with females ofCotesia marginiventris (Cresson) in Petri dishes (10 cm diam). MatedC. marginiventris females, ranging in age from 1 to 3 days exhibited the most intense bioassay responses to potential sources of kairomone. Contacting a FAW-damaged corn leaf modified the pattern of movement inC. marginiventris from random to one exhibiting a significant increase in klinokinesis. No significant differences were present in kairomone responses of nonconditioned and conditioned parasitoids and parasitoid response to kairomones did not change throughout the photophase (0800–1800 hr). Removal of one, both, or the first eight antennal segments reduced or eliminated the response of the parasitoid to kairomones. Female parasitoids did not exhibit a preference for corn leaves damaged by a particular fall armyworm instar and parasitization rates were highest in larvae 48 hr old.Submitted in partial fulfillment of the requirements for Doctor of Philosophy, Department of Entomology and Nematology, University of Florida, Gainesville.     

Sex pheromone of fall armyworm

Two components of the fall armyworm,Spodoptera frugiperda (J.E. Smith), sex pheromone, (Z)-9-dodecen-1-ol acetate (DDA) and (Z)-9-tetradecen-1-ol (TDA), were tested alone and in combination to determine their effects on male sexual response and inhibition of mating in the laboratory. The threshold response for FAW males was lower for TDA than for DDA, and males responded to TDA over a wider range of dosages. Although TDA is not attractive to FAW males in the field, this compound was highly effective in reducing mating under laboratory conditions.cooperating with the Insect Attractants, Behavior, and Basic Biology Research Laboratory, Gainesville, Florida 32604.     

Enhanced maize (Zea mays L.) pericarp browning: Associations with insect resistance and involvement of oxidizing enzymes

The kernel pericarp of a maize (Zea mays L.) inbred, Mp313E, that browns rapidly at milk stage when damaged and that is resistant toAspergillus flavus Link and the dusky sap beetleCarpophilus lugubris Murray compared to a susceptible inbred, SC212M, was examined for differing oxidizing enzymes (peroxidases) and their substrate specificity. Additional pericarp enzymes were constitutively produced by the Mp313E inbred compared to the SC212M inbred, as indicated by gel electrophoresis and isoelectric focusing. These enzymes oxidized relevent pericarp substrates such as ferulic acid. Similar results were seen with two varieties of maize containing theCh mutant gene, which brown upon senescence in that enhanced oxidation of ferulic acid was seen in homogenates of browned pericarp compared to that which was cold-shocked and did not brown. Corn powder that was browned by mixing oxidizing enzymes with corn pericarp components ferulic acid and coumaric acid were typically less preferred/more toxic to caterpillars such asHelicoverpa zea (Boddie) and sap beetles such asC. lugubris. Thus, enhanced browning of maize pericarp can promote resistance to insects and is at least partly influenced by the presence of oxidizing enzymes. This mechanism may also promote resistance to maize pathogens, including those that produce mycotoxins.     

2-hydroxy-4,7-dimethoxy-1,4-benzoxazin-3-one (N-O-Me-DIMBOA), a possible toxic factor in corn to the southwestern corn borer

The southwestern corn borer (SWCB),Diatraea grandiosella Dyar, is a major pest of corn,Zea mays L., in the southern United States. The damage to corn is caused primarily by larval feeding on leaf, ear, and stem tissues. In this study, 2-hydroxy-4,7-dimethoxy-1,4-benzoxazin-3-one (N-O-Me-DIMBOA) was identified by MS and NMR as present in corn whorl surface waxes. This compound has evidently not been isolated previously, but its glucoside has been reported in corn, wheat, andCoix lachryma. It is present in the waxes in a higher concentration than DIMBOA (2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one) and 6-MBOA (6-methoxybenzoxazolinone). It was toxic to the SWCB in a stress diet, but it was less toxic to this insect than 6-MBOA when incorporated in the standard rearing diet. Nevertheless, it may have some role in the resistance of corn to the SWCB because the total surface wax content is higher in resistant lines than in susceptible lines.Mention of a trademark, proprietary product, or vendor does not constitute a guarantee or warranty of the product by the U.S. Department of Agriculture and does not imply its approval to the exclusion of other products or vendors that may also be suitable.     

Influence of pheromone trap color and design on capture of male velvetbean caterpillar and fall armyworm moths (Lepidoptera: Noctuidae)

Three pheromone traps were evaluated in paired field trials for effectiveness in capturing wild male velvetbean caterpillar moths (VBC),Anticarsia gemmatalis Hübner, and fall armyworm moths (FAW),Spodoptera frugiperda (J.E. Smith), using a wind-oriented trapping device. The traps were: (1) the standard multicolored bucket trap consisting of a forest green canopy, yellow funnel, white bucket and open (i.e., single wire) pheromone holder; (2) a forest green monocolored bucket trap; and (3) the Multi-Pher-1 trap consisting of a blue-green canopy, white funnel, white bucket, and white, multislotted pheromone holder. The Multi-Pher-1 trap differs primarily from the other two in that its entrance is smaller in diameter and is partially obstructed by the pheromone holder. Significantly fewer VBC and FAW males were captured in pheromone-baited monocolored (forest green) bucket traps than standard multicolored bucket traps. The Multi-Pher-1 trap also caught significantly fewer VBC moths than the standard multicolored bucket trap, but there was no significant difference in numbers of FAW moths caught in the two traps. The results further demonstrate the importance of considering visual cues, in this case color, in the design of pheromone traps for nocturnal insects.This article reports the results of research only. Mention of a proprietary product does not constitute an endorsement or the recommendation for its use by USDA.     

Herbivore- and Elicitor-Induced Resistance in Rice to the Rice Water Weevil (<Emphasis Type="Italic">Lissorhoptrus oryzophilus</Emphasis> Kuschel) in the Laboratory and Field

Feeding by herbivores can change plants in ways that make them more resistant to subsequent herbivory. Such induced responses are better-studied in a number of model dicots than in rice and other cereals. In a series of greenhouse and field experiments, we assessed the effects of prior herbivory by the fall armyworm, Spodoptera frugiperda (J.E. Smith) and of exogenous applications of jasmonic acid (JA) on the resistance of rice plants to the rice water weevil, Lissorhoptrus oryzophilus (Kuschel), the major pest of rice in the United States. Prior feeding by S. frugiperda and treatment of plants with exogenous JA resulted in increases in the resistance of plants to the weevil. Increases in resistance were manifested as reduced numbers of eggs and first-instars associated with armyworm-injured or JA-treated plants relative to control plants. In field experiments, there was a transient but significant reduction in the number of immature L. oryzophilus on JA-treated plants relative to untreated plants. To our knowledge, this is the first example of direct induced resistance in rice demonstrated in small-plot field experiments. We discuss the potential for the use of elicitor induced resistance in rice.     

Mating and oviposition byCotesia (=Apanteles) marginiventris (Hymenoptera: Braconidae) in presence of synthetic pheromone ofSpodoptera frugiperda (Lepidoptera: Noctuidae)

Two blends of fall armyworm,Spodoptera frugiperda (J.E. Smith), pheromonal components were evaluated in the laboratory for possible close-range effects on mating and oviposition byCotesia marginiventris (Cresson). Neither a two- nor a five-component blend had any significant effect on mating or parasitization ofS. frugiperda larvae by this parasitoid. Dosages of 200, 400, and 800 g were tested.     

The Attraction of <Emphasis Type="Italic">Spodoptera frugiperda</Emphasis> Neonates to Cowpea Seedlings is Mediated by Volatiles Induced by Conspecific Herbivory and the Elicitor Inceptin

Neonate fall armyworms [FAW; Spodoptera frugiperda (Smith)] often encounter conspecific herbivore damage as they disperse from an egg mass to an initial feeding site. We investigated the orientation responses of dispersing neonates to herbivore damage in cowpea seedlings, specifically examining whether neonate behaviors were affected by inceptin, the primary elicitor of FAW-induced defenses in cowpea leaves. We focused on responses to damage caused by conspecific first instars, as might occur during the dispersal of siblings from an egg mass. Inceptin contents of damaging first instar FAW were controlled through their diets, with leaf-fed FAW producing inceptins in their oral secretions, and root-fed or starved FAW lacking these elicitors. In a bioassay designed to evaluate neonate dispersal off a host plant, a higher percentage of neonates remained on herbivore-induced or inceptin-treated plants than on undamaged plants, mechanically damaged plants, freshly damaged plants, or on plants damaged by FAW lacking inceptins. Further investigations of neonate responses to plant odors with a four-arm olfactometer demonstrated that neonate attraction to odors from 4-h old FAW damage was strongly dependent on previous diet of the damaging larvae. Neonates were attracted to odors from 4-h old FAW damage over odors from undamaged plants or fresh FAW damage, provided that the damaging larvae had previously ingested leaf material. In a direct comparison of odors from induced plants, plants damaged by leaf-fed FAW were as attractive as plants treated with synthetic inceptin. GC-MS analysis confirmed that (E)-4,8-dimethyl-1,3,7-nonatriene (DMNT) was the major volatile induced by FAW herbivory. While both DMNT and undamaged plant odors were more attractive than air, neonates preferred DMNT-supplemented plant odors. These results suggest that neonate FAW exploit herbivore-induced plant volatiles as host plant location and recognition cues. The use of trade, firm, or corporation names in this publication (or page) is for the information and convenience of the reader. Such use does not constitute an official endorsement or approval by the United States Department of Agriculture or the Agricultural Research Service of any product or service to the exclusion of others that may be suitable.     

In vitro metabolism of a linear furanocoumarin (8-methoxypsoralen,xanthotoxin) by mixed-function oxidases of larvae of black swallowtail butterfly and fall armyworm

Studies were made of the comparative in vitro metabolism of [¹⁴C]xanthotoxin and [¹⁴C]aldrin by homogenate preparations of midguts and bodies (carcass minus digestive tract and head) of last-stage larvae of the black swallowtail butterfly (Papilio polyxenes Fabr.) and the fall armyworm [Spodoptera frugiperda (J. E. Smith)]. The two substrates were metabolized by 10,000g supernatant microsomal preparations from both species. Evidence gained through the use of a specific inhibitor and cofactor indicated that mixed-function microsomal oxidases were major factors in the metabolism and that the specific activity of this enzyme system was considerably higher in midgut preparations fromP. polyxenes than in similar preparations fromS. frugiperda. Aldrin was metabolized 3–4 times faster byP. polyxenes, and xanthotoxin 6–6.5 times faster.     

Controlled release plastic strips containing (Z)-9-dodecen-1-ol acetate for attractingSpodoptera frugiperda

Laminated plastic strips and polyethylene vials used for dispensing (Z)-9-dodecen-1-ol acetate attracted maleSpodoptera frugiperda (J.E. Smith) in similar numbers for comparable periods. Addition of an antioxidant to the pheromone dispensed from the polyethylene vials did not significantly increase the period of effectiveness. The laminated plastic strips were acceptable dispensers of pheromone bait for attractingS. frugiperda males to sticky traps in the field.Lepidoptera: Noctuidae.This paper reports the results of research only. Mention of a commercial or proprietary product or of a pesticide in this paper does not constitute a recommendation for use by the U.S. Department of Agriculture, nor does it imply registration under FIFRA as amended.     

Effects of Peruvian Maize Extracts on Growth, Development, and Fecundity of the European Corn Borer

Twelve Peruvian maize, Zea mays, accessions were selected because of their relatively high level of field resistance to first-generation European corn borer (ECB), Ostrinia nubilalis, larval leaf-feeding. Water extracts of freeze-dried, powdered, leaf tissue were incorporated into a standard ECB diet, fed to larvae, and the effects on larval growth, development, and fecundity were measured. Larval and pupal weights were monitored as were the time elapsed in the larval, pupal, and adult stages. Adult fecundity and egg fertility were recorded. The experiment was a randomized block design (larvae and pupae) or a completely randomized design (adults) and analyzed with ANOVA ( = 0.05). Pairwise comparisons were made between groups of insects grown on diets containing extracts from the Peruvian lines, a standard diet, or diets containing extracts of a known susceptible inbred, and a known resistant inbred line. Survival was analyzed with a chi-squared test ( = 0.05). Two Peruvian accessions significantly reduced female larval and pupal weights, extended pupal and adult development time, and decreased survival of pupae and adults. Water extracts also had a pronounced impact on males; two accessions significantly reduced pupal weight and extended the time required to pupate, and one reduced male survival to adults. The results indicate that water-soluble factors from resistant Peruvian accessions inhibit the growth, developmental time, and survival of ECB. These resistance factors could be useful in the development of maize germplasm with insect-resistant traits.     

Beneficial arthropod behavior mediated by airborne semiochemicals. IX. Differential response ofTrichogramma pretiosum,an egg parasitoid ofHeliothis zea,to various olfactory cues

The behavior ofTrichogramma pretiosum Nixon wasps when exposed to different olfactory cues was studied in a wind tunnel. Compared to clean air, the sex pheromone of its hostHeliothis zea (Boddie) increased wasp residence times, walking times, and path lengths on a platform and decreased walking velocity. If wasps were released on top of a glass rod above a platform, the odor caused the wasps to land shortly after takeoff. In addition, a clear dose effect with regard to total residence and walking times was found. These responses were not elicited by three dosages of the sex pheromone ofSpodoptera frugiperda (J.E. Smith) or by a blend of saturated acetates. These results correspond with the observation thatH. zea is a common field host ofT. pretiosum, whereas eggs ofS. frugiperda are rarely attacked by this parasitoid.     

Chemical and ultrastructural analysis of corn cuticular lipids and their effect on feeding by fall armyworm larvae

The cuticular lipid composition of lower and upper leaves of five genotypes of field-grown corn,Zea mays L., was determined by combined gas chromatography-mass spectrometry. Surface lipids of the upper leaves had a higher proportion ofn-alkanes (45–52%) than the lower leaves, while the lower leaves had higher percentages of fatty alcohols (12–18%) than the upper leaves. Scanning electron microscopy showed that the upper leaves of two corn genotypes, MpSWCB-4 and Cacahuacintle X's, had a smooth amorphous appearance, while the lower leaves had a dense array of wax crystals.Spodoptera frugiperda (J.E. Smith) larvae weighed more and developed more rapidly when they were reared on diet containing corn foliage from which the cuticular lipids had been removed than when they were fed untreated foliage. However, growth was not inhibited when larvae were fed diet containing the cuticular lipid extracts or individual cuticular lipid components.     

Phytotoxin Production and Phytoalexin Elicitation by the Phytopathogenic Fungus Sclerotinia sclerotiorum

The fungus Sclerotinia sclerotiorum (Lib.) de Bary causes rot disease in a vast range of plant families, including Cruciferae (Brassicaceae). We investigated the production of phytotoxins by S. sclerotiorum by using a bioassay-guided isolation, as well as the phytoalexins produced by the resistant wild crucifer Erucastrum gallicum under elicitation by S. sclerotiorum and other agents. We established for the first time that S. sclerotiorum produces a somewhat selective phytotoxin, sclerin, which is phytotoxic to three cruciferous species (Brassica napus, B. juncea, and Sinapis alba) susceptible to Sclerotinia stem rot disease, causing severe necrosis and chlorosis, but not to a resistant species (Erucastrum gallicum). In addition, we have shown that oleic acid, the major fatty acid isolated from sclerotia of S. sclerotiorum is responsible for the toxic activity of extracts of sclerotia to brine shrimp larvae (Artemia salina). Phytoalexin elicitation in leaves of E. gallicum led to the isolation of three known phytoalexins: indole-3-acetonitrile, arvelexin, and 1-methoxyspirobrassinin. Considering that resistance of E. gallicum to S. sclerotiorum is potentially transferable to B. rapa, a susceptible canola species, and that arvelexin, and 1-methoxyspirobrassinin are not produced by B. rapa, these phytoalexins may become useful markers for resistance against S. sclerotiorum.