Spatial distribution of plants infested with Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) on corn crop

The importance of the fall armyworm, Spodoptera frugiperda (J.E. Smith), is not only due to the damage it can cause, but also to its difficult control. It is essential to know the pest population parameters, such as its standard scattering in the crop, because depending on the space arrangement of the insects in the area, different sampling methods are required. For the present work, three experimental fields were installed in different areas: the first one had 66 plots with seven plants each; the second one had 55 plots with four plants each; and the third field had 55 plots with six plants each. The presence or absence of the pest in the plant was evaluated in the samples. According to the data observed, either to the scattering index or to the theoretical scattering models, S. frugiperda dispersion is scattered at random and strongly depends on the infestation level.     

Haplotype identification within Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) corn and rice strains from Colombia

The fall army worm Spodoptera frugiperda (Smith) is a migratory important pest of corn, sorghum, rice, grass and bermudagrass in North and South America. This species has diverged into two genetically differentiated but morphologically identical strains, "the rice" and "the corn". They have been analyzed by sequencing the genes cytochrome oxydase I, II and ITS1 from populations from the United States and Brazil. However, no such studies were performed in Colombia. In here, we identified 43 haplotypes by sequencing a fragment of the COI gene from 102 individuals, of which 40 had already been identified as the "corn" and "rice" strains or to their hybrids from Tolima, and the rest were collected from corn, cotton, sorghum, grass and rice fields in other regions of Colombia. The corn strain haplotype H1 was the most frequently found in this country, representing the main target for FAW monitoring programs. AMOVA analysis confirmed the population structure between Colombian and North American S. frugiperda haplotypes (F(ST) = 0.76812, P < 0.001), but not within the different Colombian regions, suggesting high gene flow within the country. The ML trees obtained for Tolima and for Colombia as a whole did not generate clustering amongst S. frugiperda sequences, neither via host-plant association nor by geographical areas. The minimum spanning network for Colombia corroborated our finding that the haplotype H1 has the highest frequency in the country. Our data suggest that haplotype frequency determination will be useful in the establishment of a monitoring system for this species.     

Plasmid patterns of efficient and inefficient strains of Bacillus thuringiensis against Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae)

Bacillus thuringiensis harbors genes encoding Cry proteins found in chromosomes or plasmids of different sizes (4-150 Mb). Although the smaller plasmids are more abundant in B. thuringiensis, their specific function is unknown. As for the megaplasmids, their main recognized function is to harbor cry genes, although the sequencing of some of these plasmids indicates the occurrence of other important genes. This work used a new protocol for practical and rapid extraction of plasmid DNA in order to characterize the plasmid patterns of Brazilian strains belonging to Embrapa Milho e Sorgo research center B. thuringiensis bank. We tried to further assess the relationship of plasmid patterns with strains belonging to the same serovars and strains causing 100% and no mortality to Spodoptera frugiperda (J.E. Smith) larvae. It was possible to characterize 59 strains based on the migration of bands in agarose gel. Strains belonging to the same serovars showed different plasmid sizes (from 1,636 bp to 23,200 bp), with the exception of two strains belonging to serovar galleriae. The strain T09 Bt tolworthi showed a plasmid migration pattern identical to strains belonging to serovar galleriae. Plasmid patterns differed for 46 strains, confirming that this is a useful tool to discriminate specific strains. However, it was not possible to associate the plasmid pattern or the occurrence of particular plasmids with the pathogenicity of a given species towards S. frugiperda larvae.     

Oviposition, development, and reproduction of Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) fed on different hosts of economic importance

The host selection for oviposition by Spodoptera frugiperda (J.E. Smith) among corn, millet, cotton and soybean, and its relationship with the biological characteristics were investigated. Free and non-choice tests for oviposition using plots containing five plants each, from each host in plastic greenhouse, resulted in similar oviposition preference among the host plants. In addition, selected biological characteristics of S. frugiperda were determined in the laboratory with larvae feeding on host leaves, and the combination of leaf and cotton boll. Neonate larvae exhibited low success of colonization on cotton boll compared to the leaves of all other hosts. Spodoptera frugiperda fed only on cotton bolls exhibited longer larval and pupal development, and longer adult life span; however with similar egg production. Larvae fed cotton leaves during six days and then transferred to cotton bolls, however, exhibited development and reproduction similar to those reared on corn or only on cotton leaves. Therefore, the variations on immature stages of S. frugiperda were not related with host selection for oviposition which was similar among the studied hosts. Based on our data, the millet as a winter, rotational, and cover crop is a potential host for S. frugiperda, while leaves and cotton bolls were diets of intermediate suitability as compared to corn and soybean leaves.     

Association of bioassays and molecular characterization to select new Bacillus thuringiensis isolates effective against Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae)

The fall armyworm, Spodoptera frugiperda (J. E. Smith), is one of the main corn pests and Bacillus thuringiensis is important in its control because of its entomopathogenic property. The objective of this study was the molecular characterization of B. thuringiensis isolates for cry1 locus presence and the assessment of the efficiency of these isolates in controlling S. frugiperda caterpillars. Gral-cry1 was used in the PCR analyses to confirm the presence of the cry1 locus in 15 isolates. A 3 x 10(8) spore/ml suspension bathed the diet used to feed 30 caterpillars per isolate, with three replications. The cry1 locus type genes of the different isolates were identified for five gene subclasses; linear regression analyses were carried out to ascertain possible associations between the presence of an individual cry1 locus gene and high levels of toxicity. All the DNAs amplified with Gral-cry1 presented an amplification product with the expected size. Regarding the levels of insecticide efficiency against the cob worm, 41 isolates presented 100% mortality and 16 presented an index between 70% and 90%. The cry1Ab gene was present in 80 isolates, cryb in 69 isolates, cry1Ac in all the isolates and cryv and cry1E in 93 and 27 isolates, respectively. The values regarding the individual effect of each gene on caterpillar mortality were significant at 1% probability for the cry1Ac and cry1E genes.     

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

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

Toxicity of Porella chilensis Sesqui- and Diterpenoids against Larvae of the Corn Pest Spodoptera frugiperda (J.E. Smith) (Lepidotera: Noctuidae)

Porella, the largest genus of the family Porellaceae (Hepaticae) is widespread in the tropical and subtropical regions of South America. Most Porella species are rich sources of sesqui- and diterpenoids, many of which show interesting biological activities. Secondary metabolites produced by plants can interact with insects and act as antifeedants and growth regulators affecting hormone and nervous systems as well as stomach and muscle tissues. A previous chemical investigation of a Patagonian collection of Porella chilensis yielded sesqui- and diterpenoids that were now evaluated for their effects against Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae), a serious pest affecting corn crops mainly in the Americas. Four pinguisanes (1?C4), three fusicoccanes (5?C7), and one aromadendrane (8) from P. chilensis displayed larvicidal activity against S. frugiperda when incorporated to the larval diet at 100 and 200???g/g of diet with a significant decrease in the larval growing rate. The observed effects were in part produced by severe alterations of the epithelial cells of the midgut as indicated by our histological studies.     

Spatial analysis of the distribution of Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) and losses in maize crop productivity using geostatistics

The fall armyworm, Spodoptera frugiperda (J.E. Smith), is one of the chief pests of maize in the Americas. The study of its spatial distribution is fundamental for designing correct control strategies, improving sampling methods, determining actual and potential crop losses, and adopting precise agricultural techniques. In S?o Paulo state, Brazil, a maize field was sampled at weekly intervals, from germination through harvest, for caterpillar densities, using quadrates. In each of 200 quadrates, 10 plants were sampled per week. Harvest weights were obtained in the field for each quadrate, and ear diameters and lengths were also sampled (15 ears per quadrate) and used to estimate potential productivity of the quadrate. Geostatistical analyses of caterpillar densities showed greatest ranges for small caterpillars when semivariograms were adjusted for a spherical model that showed greatest fit. As the caterpillars developed in the field, their spatial distribution became increasingly random, as shown by a model adjusted to a straight line, indicating a lack of spatial dependence among samples. Harvest weight and ear length followed the spherical model, indicating the existence of spatial variability of the production parameters in the maize field. Geostatistics shows promise for the application of precise methods in the integrated control of pests.     

Influence of parasitism by Chelonus insularis Cresson (Hymenoptera: Braconidae) on the susceptibility of Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) to insecticides

The egg-larval parasitoid Chelonus insularis Cresson is a key parasitoid of the fall armyworm Spodoptera frugiperda (J. E. Smith) during the second field generation in the upper Magdalena River Basin, Colombia. In spite of selection pressure, the higher susceptibility of the second generation fall armyworm larvae to insecticides, compared with that of the first generation, suggests that the parasitism may be responsible for the apparent difference in susceptibility. Parasitized and non-parasitized 2nd-instar larvae of the fall armyworm were tested for susceptibility to chlorpyriphos, methomyl, cypermethrin, and Bacillus thuringiensis in the laboratory, using the leaf dip test. Parasitized larvae were up to 3.93 times more susceptible to chlorpyriphos, 3.71 times to methomyl, and 14.11 times to cypermethrin than non-parasitized larvae. The least effect of parasitism on susceptibility was found for B. thuringiensis. We discuss the negative influence of synthetic insecticide on the parasitoid population dynamics and its impact on insecticide resistance.     

Effects of a nucleopolyhedrovirus in mixtures with azadirachtin on Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae) larvae and viral occlusion body production

Abstract

The combined and alone effects of azadirachtin (AZA) and Spodoptera frugiperda multicapsid nucleopolyhedrovirus (SfMNPV) on the mortality of S. frugiperda (J. E. Smith) (Lepidoptera: Noctuidae) were evaluated in the laboratory. For this, diet surface contamination bioassays were performed on S. frugiperda in the third instar. LC₅₀ values for SfMNPV alone were determined to be 430 and 373 viral occlusion bodies (OBs)/mm² at 192 and 216 h after treatment, respectively. LC₅₀ values for AZA alone were estimated for two periods of continuous exposure (4 or 5 days). In this case, LC₅₀ values were 45.5 and 16.8 mg L^?1 at 216 h after treatment (4 or 5 days of larval exposition to insecticide, respectively). We observed that although the interaction of AZA with SfMNPV increased viral pathogenicity, such improvements were of greater magnitude and more consistent at the lower OB concentration used (177 OBs/mm²). Application of SfMNPV (430 OBs/mm²), AZA (26.4 mg L^?1) or SfMNPV–AZA mixtures resulted in a significant reduction in the mean weight of larvae treated in the third instar across the experiment, by 23–41%, 17–95% and 26–97%, respectively, compared to control. The duration of larval development during the third and fourth instars increased significantly in larvae exposed to SfMNPV–AZA mixtures and AZA alone compared to SfMNPV alone and control treatments. The yield of OBs/mg weight of larvae treated with SfMNPV alone was 1.8-fold higher than OB yields from insects inoculated with SfMNPV–AZA mixtures. We conclude that AZA + SfMNPV mixtures are unlikely to be useful for the mass production of this virus and laboratory observations on the value of AZA + SfMNPV mixtures as a potentiator of biological insecticides require validation in field studies under commercial growing conditions.     

Lethal and sublethal effects of methoxyfenozide on the development, survival and reproduction of the fall armyworm, Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae)

The lethal and sublethal effects of the ecdysone agonist methoxyfenozide on the fall armyworm, Spodoptera frugiperda (J. E. Smith), were investigated by feeding a methoxyfenozide-treated diet to fifth instars until pupation in doses corresponding to the LC10 and LC25 for the compound. Larval mortality reached 8% and 26% in the low and high concentration groups, respectively, on the seventh day of the experiment. A progressive larval mortality of 12% for the LC10 and 60% for the LC25 was observed before pupation. Treated larvae exhibited lower pupal weights, higher pupal mortality, presence of deformed pupae, and more deformed adults than untreated larvae. The incorporation of methoxyfenozide into the diet had a significant effect on the timing of larval development. The development period for males and females was about seven days longer than the controls for both concentrations tested. In contrast, the compound affected neither pupae nor adult longevity. Finally, S. frugiperda adults that resulted from fifth instars treated with methoxyfenozide were not affected in their mean cumulative number of eggs laid per female (fecundity), nor percentages of eggs hatched (fertility), or the sex ratio. Our results suggest that the combination of lethal and sublethal effects of methoxyfenozide may have important implications for the population dynamics of the fall armyworm.     

Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) eggs as alternative food for rearing of lady beetles Eriopis connexa (Germar) (Coleoptera: Coccinellidae)

Eriopis connexa (Germar) (Coleoptera: Coccinellidae) is an important predator with potential for biological control of insect pests. This research evaluated the development of E. connexa larvae fed on fresh eggs of Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) without (T1) or with (T2) scales or one-day (T3) or six-month (T4) frozen, or newly-hatched larvae of S. frugiperda (T5). The percentage of E. connexa adults was higher when larvae feeding on fresh S. frugiperda eggs with or without scales, or one-day frozen eggs of this prey and lower with eggs of this Lepidoptera after frozen for six months or with newly-hatched larvae of S. frugiperda. Duration of the larval period of E. connexa was 15.7, 15.8, 16.0, 17.6, and 17.3 days, respectively, with these diets. The high survival of E. connexa fed with eggs of S. frugiperda shows the potential use of this prey in the laboratory to maintain this natural enemy.     

Oviposition Choice of Two Fall Armyworm (Lepidoptera: Noctuidae) Host Strains

Fall armyworm, Spodoptera frugiperda (J. E. Smith), is a noctuid species that is composed of two morphologically identical sympatric host strains (corn and rice) that differ in their distribution among plant hosts. In an effort to explain observations of host fidelity in the field, ovipositional preference of the two strains on corn (Zea mays L.) or pasture grass (Cynodon nlemfuensis Vanderyst var. nlemfuensis) was determined using two greenhouse bioassays. In the first bioassay, corn strain females placed more eggmasses on the screen enclosure than on corn plants while grass plants contained an intermediate number of eggmasses. Rice strain females placed most of their eggmasses on grass plants. In the second bioassay, corn strain females placed an equal number of eggmasses on corn and grass plants in comparison to rice strain females which placed >3.5× more eggmasses on grass plants than on corn plants. Individual eggs as part of the eggmasses were also counted on plants and on the screen enclosure. Corn strain females equally placed eggs on the two host plants and on the screen enclosures, however rice strain females placed more eggs on grass plants compared to corn plants or the screen enclosure. This is the first report of consistent differential oviposition between corn and rice strain fall armyworm females.     

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

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

Behavior of Trichogramma atopovirilia Oatman & Platner and T. pretiosum Riley (Hymenoptera: Trichogrammatidae) on Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae) egg masses.

In this study we analyzed the impact of physical barriers of Spodoptera frugiperda (J. E. Smith) egg-masses on the behavior of Trichogramma atopovirilia Oatman & Platner and Trichogramma pretiosum Riley. The duration of drumming, drilling, oviposition, period spent over the egg-mass, and interval between parasitized eggs were timed, and the number of parasitized eggs were recorded. The presence of scales on the egg-masses caused a significant increase in the time spent by both parasitoids on each process and a decrease in the residence time over the egg-mass and in the number of parasitized eggs, with an increase in the number of egg layers. There was a significant decrease in the number of parasitized eggs in relation to egg-masses with one layer and no scales. We observed that the physical barriers in fall armyworm egg-masses changed the behavior of T. atopovirilia and T. pretiosum, affecting their parasitization capacity.     

Juvenile hormone biosynthesis in the fall armyworm, Spodoptera frugiperda (Lepidoptera, Noctuidae)

The in vitro production of juvenile hormones (JH) was investigated by using corpora allata (CA) of larvae and corpora cardiaca-corpora allata (CC-CA) complexes of adult females of the fall armyworm Spodoptera frugiperda. In female moths, JH release was high compared to that in 5th and 6th instar larvae. Concentrations of 0.11-0.12 mM methionine, 180-200 mM Na(+), 5.8-8.3 mM K(+), 10-50 mM Ca(2+) and a pH range of 5.7-6.3 yielded optimal incorporation of L-[methyl-(3)H] methionine in vitro by CC-CA complexes. The highest hourly incorporation occurred during a 9-h incubation period following a 1.5-h lag-phase. JH release from CC-CA complexes of adult females was shown to be age-dependent with a peak value on day 2 (approx. 4 pmol h(-1) CA(-1)). By a combination of reversed phase (RP)- and normal phase (NP)-high performance liquid chromatography (HPLC), two major labelled products released by the complex were separated. One compound co-migrated with chemically synthesized JH II diol, the second compound with JH III diol. Only traces of JH II and III could be detected in some samples. Gland extracts also contained both the major radiolabelled products. Double labelling experiments using [3H]methionine and [14C]acetate confirmed their de novo synthesis in CC-CA complexes of female moths. The nature of chemically synthesized reference JH III diol was proved by LC-MS (ESI mass spectrometry) and 1H-NMR (nuclear magnetic resonance spectroscopy).     

Biochemical Analysis of a Castor Bean Leaf Extract and its Insecticidal Effects Against Spodoptera frugiperda (Smith) (Lepidoptera: Noctuidae)

Plant extracts represent a great source of molecules, with insecticidal activity, which are used for pest control in several crop production systems. This work aimed to evaluate the toxicity of an aqueous extract of leaves of castor bean against larvae of Spodoptera frugiperda (Smith) (Lepidoptera: Noctuidae) in search for different classes of molecules with insecticidal activities by using in vitro assays. The effects of the castor bean leaf extract on the food utilization, development, and survival of S. frugiperda larvae was evaluated by feeding the larvae an artificial diet supplemented with different concentrations of the extract (0%, 1%, 2.5%, 5%, and 10% w/v). The effects observed were dose-dependent, and the highest concentration evaluated (10% w/v) was the one the most affected food utilization by altering the nutritional indices, as well as larval weight gain, development time, and survivorship. In vitro assays to detect saponins, lectins, and trypsin inhibitors in the castor bean leaf extract were performed, but only trypsin inhibitors were detected. No preference for the diet source was detected in S. frugiperda by feeding the larvae in choice experiments with diets containing different concentrations of the castor bean extract tested. The data obtained indicate the existence of a potential molecule in the tested extract of castor bean to be used as an alternative insecticide to be integrated in the management of S. frugiperda.     

Host plant association and genetic differentiation of corn and rice strains of Spodoptera frugiperda Smith (Lepidoptera: Noctuidae) in Colombia

Spodoptera frugiperda (Smith) is a polifagous insect of major economic impact in the western hemisphere and exhibits two strains (i.e., corn and rice) that are morphologically identical but differ in ecology, genetics and physiology. In this work we identified these strains and their respective hybrids by using a PCR-RFLP of the COI gene and PCR of the tandem region FR. Moreover, we performed a population structure analysis by using 253 larvae from Tolima, a region where S. frugiperda is a pest on corn, rice, sorghum and cotton. Corn strain was found on 42% in corn, 34% in cotton, 19% in sorghum and 0.04 % in rice and rice strain on 35% in corn, 0.06% in cotton, 0.06% in sorghum and 53% in rice, demonstrating that corn strain specificity is superior to rice strain. Hybrids between these strains were more abundant in corn. The distributions on their host plants reflect a population genetic differentiation in S. frugiperda with values of PhiPT (COI) = 0.31, P < 0.0001, PhiPT (FR) = 0.17, P < 0.0001 for all crops and PhiPT (COI) = 0.42, P < 0.01, PhiPT (FR) = 0.13, P < 0.01 for the sixteen sampled farms. The dendrograms showed two clusters representing both strains. The results obtained in this study suggest that the management of this insect must differ on each host plant, given the specialization that both strains present, particularly in corn and rice.     

Assortative Mating and Lack of Temporality Between Corn and Rice Strains of Spodoptera frugiperda (Lepidoptera,Noctuidae) from Central Colombia

Spodoptera frugiperda is a Neotropical moth that has diverged into “corn” and “rice” strains. In the US, prezygotic isolation studies have shown that both populations mate assortatively. In addition, recent studies have demonstrated that mating by the same strain individuals is enhanced by an allochronic shift in mating activity and male pheromones are important during courtship. In Colombia, studies on mate choice have never been performed previously, although earlier analyses made in populations from Central Colombia showed no significant differences in time of first mating and copula duration between the strains. Here, we performed multiple choice experiments using a tetrad design composed of individuals of the corn and the rice strains. We found that corn strain females rarely mate with rice strain males, but rice strain females mate with both strains of males. In addition, no temporal isolation was found. A ML (maximum likelihood) approach was used to discriminate between mating propensity and mate choice behaviors in S. frugiperda strains. This approach showed that mating propensity of corn strain males is three times greater than rice strain males. In contrast, in females, the propensity of mating was slightly higher for the corn strain. Finally, the isolation index between the corn and the rice strains from Colombia produced a value of I?=?0.33. Our results suggest that prezygotic isolation at the behavioral and temporal levels differ between the US and Colombia. Moreover, in the US temporal isolation appears to have an important role in behavioral isolation but in Colombia temporality is not necessary to reduce encounters between S. frugiperda strains.     

Circadian Locomotor Activity in the Larva and Adult Fall Armyworm, Spodoptera frugiperda (Noctuidae): Effect of Feeding with the Resistant Variety of Maize CML67

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 artificial light-dark conditions, suggesting that the maize variety CML67 may possess some active substance(s) that affect the maturation of the circadian system, controlling locomotor activity rhythms in larvae and adult armyworms.