Corn Earworm Larvae Research Articles

ENTOMOPATHOGENIC NEMATODES ASSOCIATION WITH SOIL- DWELLING BACTERIA: THE CASE OF STEINERNEMA FELTIAE

The symbiosis of entomopathogenic nematodes (EPNs) with bacteria in the genera Photorhabdus and Xenorhabdus is well known. However, other soil-dwelling bacteria can occasionally be isolated from EPNs collected from diseased larvae. Accordingly, through several studies, we have detected the presence of diverse bacterial species including isolates of Pseudomonas protegens and Serratia spp., in the body of different strains of the entomopathogenic nematode Steinernema feltiae. After documenting the presence of such bacteria inside the nematodes, both nematodes and EPN-associated bacteria were cultured, and bioassays were conducted to determine their potential against different insect targets. An isolate of P. protegens caused over 60% and 90% mortality of house fly and corn earworm larvae, respectively. These insecticidal effects were dose-dependent. The presence of the bacterium confers virulence to the nematode. According to our studies, it appears that these non-core bacteria can establish occasional or stable associations with S. feltiae. Key Words: Biological control, EPN, bacteria, symbiosis, Pseudomonas, soil, bioinsecticide.

Competition between brown stink bug (Hemiptera: Pentatomidae) and corn earworm (Lepidoptera: Noctuidae) in field corn.

Interspecific competition is an important ecological concept which can play a major role in insect population dynamics. In the southeastern United States, a complex of stink bugs (Hemiptera: Pentatomidae), primarily the brown stink bug, Euschistus servus (Say), and corn earworm, Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae), are the 2 most common pests of field corn, Zea mays L. (Poales: Poaceae). Stink bugs have the greatest potential for economic injury during the late stages of vegetative corn development when feeding can result in deformed or "banana-shaped" ears and reduced grain yield. Corn earworm moths lay eggs on corn silks during the first stages of reproductive development. A 2-year field study was conducted to determine the impact of feeding by the brown stink bug during late-vegetative stages on subsequent corn earworm oviposition, larval infestations, and grain yield. Brown stink bug feeding prior to tasseling caused deformed ears and reduced overall grain yield by up to 92%. Across all trials, varying levels of brown stink bug density and injury reduced the number of corn earworm larvae by 29-100% and larval feeding by 46-85%. Averaged across brown stink bug densities, later planted corn experienced a 9-fold increase in number of corn earworm larvae. This is the first study demonstrating a competitive interaction between these major pests in a field corn setting, and these results have potential implications for insect resistance management.

Influence of varieties of hemp, Cannabis sativa (Rosales: Cannabaceae), and fertilization rates on damage caused by corn earworm, Helicoverpa zea (Lepidoptera: Noctuidae).

Industrial hemp, Cannabis sativa L., production has been negatively impacted by larvae of corn earworm, Helicoverpa zea (Boddie), which feed on developing inflorescences. Adult H. zea oviposit on hemp once flowers develop, and late-instar larvae can cause serious loss to both quality and yield. A 2-year study to assess the influence of hemp variety and fertilization practices on damage caused by H. zea was conducted. Differences in damage ratings among varieties were observed in both years; however, the rate of nitrogen applied did not influence biomass yield or damage rating. These results indicate that increasing nitrogen fertility may not be an effective means of cultural control for mitigating damage from H. zea. Floral maturity was very influential on damage caused by H. zea as late-maturing varieties had much less floral injury than those which matured early in outdoor field trials. Some cannabinoids were also correlated to damage rating, but this relationship was due to late-maturing plants with immature flowers low in cannabinoid concentrations receiving less floral injury. Based on these results, the selection of high-yielding varieties that flower when ovipositional activity of H. zea is expected to decline should be the first step in an integrated pest management program for hemp production. This research expanded our knowledge of the role of fertility rate, varietal characteristics, cannabinoid profile, and floral maturity on damage caused by H. zea to hemp. Findings from this research will allow growers to make more informed agronomic decisions before planting to improve hemp production.

Intestinal Bacterial Diversity and Functional Analysis of Three Lepidopteran Corn Ear Worm Larvae

Simple SummaryCorn is one of the most important food crops in the world and comprises a large proportion of crops in China. Pests are one of the important factors affecting corn yield. Conogethes punctiferalis, Ostrinia furnacalis and Helicoverpa armigera are three main pests in the ear stages of corn, which significantly affect the yield and quality of corn. The three co-occurring lepidopteran pests at the ear stage occur frequently. Recently, the amount of Conogethes punctiferalis increased, even becoming the most serious pest in the Huang-Huai-Hai summer corn region of China, which is the second-largest corn producing area in China. Gut bacteria play important roles in insect adaptation to various environments. This study aimed to compare the diversity and function of the intestinal bacteria in three co-occurring lepidopteran pests, and to explore the reason for their prevalence. The results might provide insight into the prevalence of corn earworm larvae from the perspective of gut microbiota and function prediction.Insects, as the most abundant animal group on earth, and their symbionts help their hosts to adapt to various environments. Conogethes punctiferalis, Ostrinia furnacalis and Helicoverpa armigera are three main pests co-occurring in the ear stage of corn, which significantly affect the yield and quality of corn. The purpose of this study was to compare the diversity and function of the intestinal bacteria of the three co-occurring lepidopteran pests, C. punctiferalis, O. furnacalis and H. armigera, and to explore the reason of their prevalence from the microbiota’s view. Our results showed the difference of diversity and abundance of the gut bacteria of three co-occurring lepidopteran pests at the ear stage. Proteobacteria and Firmicutes were the dominant phyla, and the Enterobacteriaceae and Enterococcaceae were the dominant families in the three pests. Compared with the other two pests, Bacteroidetes was found much more in C. punctiferalis. In addition, C. punctiferalis showed more correlation and similarity in bacteria composition with corn endophytic bacteria, as well as had obvious advantages in metabolic, environmental information processing, cellular processes and organic systems function pathways. Our findings may provide insight into the prevalence of corn earworm larvae from the perspective of gut microbiota and function prediction.

Parasitism of Corn Earworm, Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae), by Tachinid Flies in Cultivated Hemp.

Simple SummaryHemp has become a rapidly growing industry in the United States in recent years. However, due to many decades of prohibition, there has been relatively little research on insect pests and their interactions with natural enemies in hemp production systems. Here, we provide the first quantitative assessment of corn earworm (CEW) Helicoverpa zea parasitism in a hemp system. Corn earworm larvae exhibited high parasitism rates by tachinid flies resulting in elevated mortality. Host mortality increased with the number of tachinid eggs per larva even though typically only one parasitoid successfully developed per host. Larger CEW larvae were more likely to survive parasitism, but frequently, neither parasitoid nor host larvae successfully developed. Our results suggest that tachinid flies hold promise as biological control agents for populations of this important pest attacking hemp.In a survey on hemp grown in western Kentucky we found an average of 27.8 CEW larvae per plant. We recorded 45% parasitism of CEW in these fields by two species of tachinid flies, Winthemia rufopicta and Lespesia aletiae. Most parasitized larvae were third to sixth instars at the time of collection. We found up to 22 tachinid eggs per host larva, 89% of which typically bore between 1 and 5 eggs on the thorax. 45.9% of CEW bearing eggs died. The number of tachinid eggs per host was unrelated to host body mass, but both the number of tachinid eggs and caterpillar body mass influenced CEW survival. Larger CEW often survived parasitism and the number of fly eggs was negatively related to survival rate. The emergence of adult flies was positively correlated with the number of eggs, but no influence of the host size was found. High mortality of CEW larvae and the parasitoids developing within them in this system suggests that secondary chemicals (or poor nutrition) of the hemp diet may be negatively affecting host and parasitoid development and influencing their interactions.

Timely Application of Four Insecticides to Control Corn Earworm and Fall Armyworm Larvae in Sweet Corn.

Simple SummaryCorn earworm and fall armyworm are the most important Lepidopteran pests in sweet corn worldwide. Timely application of insecticides during pollination is crucial to control both species and produce kernels with higher quality. Our results showed that applications of insecticides should be conducted at 48 h after pollination. Emamectin benzoate and chlorantraniliprole provided an adequate control of fall armyworm and corn earworm larvae. Furthermore, emamectin benzoate should be applied to larvae in later instars (fourth–sixth) while chlorantraniliprole and spinetoram should be sprayed when infestations of larvae in earlier instars (first–third) are observed.Insecticide sprays are a common practice to control corn earworm, Helicoverpa zea (Boddie), and fall armyworm, Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae), in corn (Zea mays L.) at reproductive stages. Our objectives were to determine (1) the most appropriate time for insecticide applications and (2) the effect of four insecticides on the survival of larvae as well as their weight. ß-cyfluthrin (0.4 mL/L), chlorantraniliprole (0.6 mL/L), emamectin benzoate (0.2 g/L), and spinetoram (1.5 mL/L) were sprayed on silks of sweet corn planted in Isabela and Lajas, Puerto Rico 3 h before and 24 and 48 h after pollination. The number of kernels produced and the damage of larvae on kernels were quantified at harvest. In addition, percentages of mortality and changes on larval weight were noted at 96 h after insecticide applications. Insecticide sprays at 3 h before pollination reduced the number of kernels or were similar to the control in all treatments. However, emamectin benzoate sprayed in Lajas and chlorantraniliprole applied in Isabela at 48 h after pollination increased the number of kernels (281–294) and reduced the damage of larvae on kernels (<0.5%) compared to the control (201–229; >7%). Furthermore, applications of emamectin benzoate caused higher percentages of fall armyworm larval mortality (>70%). Conversely, ß-cyfluthrin and chlorantraniliprole caused lower percentages of mortality (<30%) and only chlorantraniliprole and spinetoram reduced the weight of corn earworm and fall armyworm larvae collected in both locations. This information may help pest management programs and corn breeders to schedule insecticide sprays and pollination in the field.

Neonicotinoids can cause arrested pupal ecdysis in Lepidoptera

Recently, we reported a novel mode of action in monarch butterfly (Danaus plexippus) larvae exposed to neonicotinoid insecticides: arrest in pupal ecdysis following successful larval ecdysis. In this paper, we explore arrested pupal ecdysis in greater detail and propose adverse outcome pathways to explain how neonicotinoids cause this effect. Using imidacloprid as a model compound, we determined that final-instar monarchs, corn earworms (Helicoverpa zea), and wax moths (Galleria mellonella) showed high susceptibility to arrested pupal ecdysis while painted ladies (Vanessa cardui) and red admirals (Vanessa atalanta) showed low susceptibility. Fall armyworms (Spodoptera frugiperda) and European corn borers (Ostrinia nubilalis) were recalcitrant. All larvae with arrested ecdysis developed pupal cuticle, but with incomplete shedding of larval cuticle and unexpanded pupal appendages; corn earworm larvae successfully developed into adults with unexpanded appendages. Delayed initiation of pupal ecdysis was also observed with treated larvae. Imidacloprid exposure was required at least 26 h prior to pupal ecdysis to disrupt the molt. These observations suggest neonicotinoids may disrupt the function of crustacean cardioactive peptide (CCAP) neurons, either by directly acting on their nicotinic acetylcholine receptors or by acting on receptors of inhibitory neurons that regulate CCAP activity.

Presence of Corn Earworm and Fall Armyworm (Lepidoptera: Noctuidae) Populations in Sweet Corn and their Susceptibility to Insecticides in Puerto Rico

Corn earworm, Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae), and fall armyworm, Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae), are important pests in sweet corn. Our objectives were to assess the occurrence of the Lepidopteran species affecting sweet corn in Puerto Rico, and to evaluate the efficacy of 9 insecticides to control larvae of corn earworm and fall armyworm. Spodoptera frugiperda was observed in all plant stages, whereas H. zea and Diatraea saccharalis (F.) (Lepidoptera: Crambidae) affected only ears. Larvae of corn earworm and fall armyworm were susceptible (mortality > 80% at 96 h) to Steinernema carpocapsae (Weiser) (Nematoda: Steinernematidae) + oil and to methomyl, respectively, whereas both species were susceptible to chlorpyrifos. The LC50 values for chlorpyrifos was 248 ppm, whereas 312,500 S. carpocapsae nematodes per L + 625 ppm of rapeseed oil caused 53% of larval mortality at 120 h post-treatment for corn earworm larvae. Sumario El gusano de la mazorca del maiz, Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae), y el gusano cogollero, Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae), son plagas de importancia economica en el maiz dulce. Los objetivos de esta investigacion fueron identificar las especies de Lepidopteros que afectan el maiz dulce y evaluar la eficacia de 9 insecticidas para el control de las larvas del gusano de la mazorca y el gusano cogollero. Spodoptera frugiperda fue observado en todos los estados fenologicos del maiz mientras que H. zea y Diatraea saccharalis (F.) (Lepidoptera: Crambidae) afectaron solo las mazorcas. Las larvas del gusano de la mazorca y cogollero fueron susceptibles (mortalidad sobre el 80%) a Steinernema carpocapsae (Weiser) (Nematoda: Steinernematidae) + aceite y methomyl, respectivamente; mientras que las 2 especies fueron susceptibles a chlorpyrifos. Las CL50 para chlorpyrifos fue de 248 ppm, mientras que 312,500 nematodos de S. carpocapsae por L + 625 ppm de aceite de colza causo 53% de mortalidad en larvas a las 120 h despues de la aplicacion para el gusano de la mazorca. Key Words: Helicoverpa zea (Boddie); larval mortality; lethal concentrations; Spodoptera frugiperda (J. E. Smith); vegetative and reproductive stages View this article in BioOne https://doi.org/10.1653/024.102.0228

Development and application of a quantitative bioassay to evaluate maize silk resistance to corn earworm herbivory among progenies derived from Peruvian landrace Piura.

Corn earworm (CEW), Helicoverpa zea (Boddie), (Lepidoptera: Noctuidae), is a major insect pest of corn (Zea mays spp. mays L.). CEW larvae feed on silks, kernels and cobs, causing substantial yield and quality losses both through herbivory and by vectoring pathogens. The long-term goal of this work is to elucidate the genetic and biochemical basis of a potentially novel CEW resistance source discovered in silk tissue of Piura 208, a Peruvian landrace of maize (PI 503849). We developed a quantitative CEW bioassay and tested it on four populations that contrast alleles from Piura 208 with those from GT119, a CEW-susceptible maize inbred line. In replicated analyses of two populations of F1:2 families, corn genotype accounts for 84% and 68% of the variance in CEW larval weights, and up to 60% of the variance in CEW pupation percentage, demonstrating both the success of the quantitative bioassay and the strength of the Piura 208 resistance mechanism. Analyses of two corresponding populations of BC1:2 families revealed substantially diminished effects of corn genotype on CEW weight gain and pupation. This loss of Piura 208-derived CEW resistance during backcrossing suggests complex (multi-genic) inheritance of a threshold-dependent mechanism. Technical factors in bioassay performance were also assessed, often by analyzing the 1,641 CEW larvae that were raised on control diet (meridic with no corn silks added). Minor, but statistically significant impacts on CEW weight gain, pupation, and mortality were attributable to multiple technical factors in the preparation, incubation and evaluation phases of the bioassay, demonstrating the importance of randomization, stratification, replication, and variable-tracking across the many steps of this quantitative CEW bioassay. Overall, these findings indicate that this scaled-up, quantitative CEW bioassay is fundamentally sound and that Piura 208-derived resistance alleles are experimentally tractable for genetic and mechanistic research using this approach.

Simple Methods to Devitalize Eggs and Larvae ofHeliothis virescensandHelicoverpa zeaunder Laboratory Conditions

Heliothine moths are economically damaging insect pests, significantly affecting world crop production each year. Several species in the subfamily are commonly used in agricultural research laboratories. Research protocols should include methods to ensure that excess individuals are completely devitalized before being discarded, preventing further spread of the pests. Processes such as ultracold temperature freezing, autoclaving, or holding insects under adverse conditions for a long period of time are commonly used devitalization methods. Each method adds significant amounts of time and cost; also increasing competition for space in already crowded laboratories. While these frequently used methods might be necessary for some very resilient species, other species can succumb under less elaborate methods of devitalization. One hour of exposure to undiluted household bleach, brief exposure (>20 seconds) of wetted individuals to microwave radiation, storage in a household freezer for 48 hours, and contact with 70% isopropyl alcohol for 24 hours completely devitalized eggs and L1 larvae of corn earworm, Helicoverpa zea (Boddie), and tobacco budworm, Heliothis virescens (Fab.). These very effective methods might also be applicable for other noctuid species.

Impact of Corn Earworm (Lepidoptera: Noctuidae) on Field Corn (Poales: Poaceae) Yield and Grain Quality.

Corn earworm, Helicoverpa zea (Boddie), commonly infests field corn, Zea mays (L.). The combination of corn plant biology, corn earworm behavior in corn ecosystems, and field corn value renders corn earworm management with foliar insecticides noneconomical. Corn technologies containing Bacillus thuringiensis (Bt) Berliner (Bacillales: Bacillaceae) were introduced that exhibit substantial efficacy against corn earworm and may reduce mycotoxin contamination in grain. The first generation Bt traits in field corn demonstrated limited activity on corn earworm feeding on grain. The pyramided corn technologies have greater cumulative protein concentrations and higher expression throughout the plant, so these corn traits should provide effective management of this pest. Additionally, reduced kernel injury may affect physical grain quality. Experiments were conducted during 2011-2012 to investigate corn earworm impact on field corn yield and grain quality. Treatments included field corn hybrids expressing the Herculex, YieldGard, and Genuity VT Triple Pro technologies. Supplemental insecticide treatments were applied every 1-2 d from silk emergence until silk senescence to create a range of injured kernels for each technology. No significant relationship between the number of corn earworm damaged kernels and yield was observed for any technology/hybrid. In these studies, corn earworm larvae did not cause enough damage to impact yield. Additionally, no consistent relationship between corn earworm damage and aflatoxin contamination was observed. Based on these data, the economic value of pyramided Bt corn traits to corn producers, in the southern United States, appears to be from management of other lepidopteran insect pests including European and southwestern corn borer.

Residual and Systemic Efficacy of Chlorantraniliprole and Flubendiamide Against Corn Earworm (Lepidoptera: Noctuidae) in Soybean.

Experiments were conducted in Mississippi from 2013 to 2015 to determine the systemic and residual efficacy of chlorantraniliprole and flubendiamide against corn earworm, Helicoverpa zea (Boddie), in soybean. Both insecticides were applied at V4 and R3. Ten leaves that were present at the time of application and 10 newly emerged leaves that were not present at the time of application were collected to measure residual and systemic efficacy, respectively. Ten pods were removed from each plot at R5.5. For all assays, corn earworm larvae were placed on plant material. Chlorantraniliprole appeared to provide systemic control of H. zea, but was dependent on soybean growth stage at the time of application. In the V4 experiment, chlorantraniliprole resulted in greater mortality than the control on new leaves at 7 d after treatment, but not at 14 d. In the R3 experiment, chlorantraniliprole resulted in greater than 90% mortality on new leaves at all evaluation intervals. Mortality of H. zea on new leaves was <17% for flubendiamide and was not different than the control. Both insecticides resulted in significant mortality of H. zea on leaves that were present at the time of application for at least 31 d after application. Chlorantraniliprole resulted in greater mortality than flubendiamide at 24 and 31 d. Neither insecticide resulted in mortality of H. zea feeding on reproductive structures. These results suggest that chlorantraniliprole moves to new vegetative structures but not to reproductive structures of soybean, and that flubendiamide does not move systemically.

Three sorghum serpin recombinant proteins inhibit midgut trypsin activity and growth of corn earworm

The sorghum (Sorghum bicolor) genome contains a number of putative serpins (serine protease inhibitors) that are expressed under varying conditions, but little is known about their biological function. One of the sorghum serpin genes encodes a protein that contains reactive center residues Leu-Arg-X (X=small residue), called a LR serpin. Similar plant-derived LR serpin proteins can inactivate mammalian trypsins and have activity against insect trypsins. In this study the sorghum LR serpin, and two non-LR sorghum serpins, which were expressed in Escherichia coli and purified using immobilized metal-affinity chromatography, were shown to inhibit in vitro trypsin activity from larval midgut extract of corn earworm (Helicoverpa zea) and fall armyworm (Spodoptera frugiperda). Each serpin was added individually to sorghum leaf insect diet that was fed to corn earworm and fall armyworm larvae. Statistically significant reductions (30–53%) in the mean weight of corn earworm larvae, but not for fall armyworm larvae, were found in the larvae feeding on diet containing each of the serpins compared to the mean weight of those feeding on control diet. These studies suggest that the sorghum serpin genes could be utilized for corn earworm larvae resistance in sorghum breeding but fall armyworm larvae have compensatory mechanisms to counter the tested sorghum serpins.

Multifaceted effects of host plants on entomopathogenic nematodes

The success of parasites can be impacted by multi-trophic interactions. Tritrophic interactions have been observed in parasite-herbivore-host plant systems. Here we investigate aspects of multi-trophic interactions in a system involving an entomopathogenic nematode (EPN), its insect host, and host plant. Novel issues investigated include the impact of tritrophic interactions on nematode foraging behavior, the ability of EPNs to overcome negative tritrophic effects through genetic selection, and interactions with a fourth trophic level (nematode predators). We tested infectivity of the nematode, Steinernema riobrave, to corn earworm larvae (Helicoverpa zea) in three host plants, tobacco, eggplant and tomato. Tobacco reduced nematode virulence and reproduction relative to tomato and eggplant. However, successive selection (5 passages) overcame the deficiency; selected nematodes no longer exhibited reductions in phenotypic traits. Despite the loss in virulence and reproduction nematodes, first passage S. riobrave was more attracted to frass from insects fed tobacco than insects fed on other host plants. Therefore, we hypothesized the reduced virulence and reproduction in S. riobrave infecting tobacco fed insects would be based on a self-medicating tradeoff, such as deterring predation. We tested this hypothesis by assessing predatory success of the mite Sancassania polyphyllae and the springtail Sinella curviseta on nematodes reared on tobacco-fed larvae versus those fed on greater wax moth, Galleria mellonella, tomato fed larvae, or eggplant fed larvae. No advantage was observed in nematodes derived from tobacco fed larvae. In conclusion, our results indicated that insect-host plant diet has an important effect on nematode foraging, infectivity and reproduction. However, negative host plant effects, might be overcome through directed selection. We propose that host plant species should be considered when designing biocontrol programs using EPNs.

Effect of Native Mexican Isolates ofIsaria fumosorosea(Wize) Brown &amp; Smith1onSpodoptera exigua(Hübner) andHelicoverpa zea(Boddie)2

Abstract. The aim of this study was to evaluate pathogenicity and effect of four isolated native and a strain of Isaria fumosorosea on beet armyworm, Spodoptera exigua (Hubner), and corn earworm, Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae). Bioassays in a laboratory used neonate larvae of beet armyworm and corn earworm reared on artificial diet, which were inoculated by spraying or immersing with suspensions of 1 × 108 ml-1 conidia of I. fumosorosea, and subsequently incubated at 26°C, 65 ± 5% relative humidity, and 14:10 light: dark hours. Isolated HIB-30 killed the most immersed corn earworms, but fewest beet armyworms. HIB-30 most disrupted metamorphosis, 40%, by immersing beet armyworms, while HIB-29 and Pfr-612 least interrupted metamorphosis (8%), by spraying and immersing, respectively.

The Impact of Inter-Kernel Movement in the Evolution of Resistance to Dual-Toxin Bt-Corn Varieties in Helicoverpa zea (Lepidoptera: Noctuidae).

Seeds or kernels on hybrid plants are primarily F(2) tissue and will segregate for heterozygous alleles present in the parental F(1) hybrids. In the case of plants expressing Bt-toxins, the F(2) tissue in the kernels will express toxins as they would segregate in any F(2) tissue. In the case of plants expressing two unlinked toxins, the kernels on a Bt plant fertilized by another Bt plant would express anywhere from 0 to 2 toxins. Larvae of corn earworm [Helicoverpa zea (Boddie)] feed on a number of kernels during development and would therefore be exposed to local habitats (kernels) that varied in their toxin expression. Three models were developed for plants expressing two Bt-toxins, one where the traits are unlinked, a second where the traits were linked and a third model assuming that maternal traits were expressed in all kernels as well as paternally inherited traits. Results suggest that increasing larval movement rates off of expressing kernels tended to increase durability while increasing movement rates off of nonexpressing kernels always decreased durability. An ideal block refuge (no pollen flow between blocks and refuges) was more durable than a seed blend because the refuge expressed no toxins, while pollen contamination from plants expressing toxins in a seed blend reduced durability. A linked-trait model in an ideal refuge model predicted the longest durability. The results suggest that using a seed-blend strategy for a kernel feeding insect on a hybrid crop could dramatically reduce durability through the loss of refuge due to extensive cross-pollination.

Identification of a Bioactive Bowman–Birk Inhibitor from an Insect-Resistant Early Maize Inbred

Breeding of maize, Zea mays, has improved insect resistance, but the genetic and biochemical basis of many of these improvements is unknown. Maize oligonucleotide microarrays were utilized to identify differentially expressed genes in leaves of three maize inbreds, parents Oh40B and W8 and progeny Oh43, developed in the 1940s. Oh43 had enhanced leaf resistance to corn earworm larvae, Helicoverpa zea, and fall armyworm larvae, Spodoptera frugiperda, compared to one or both parents. Among ca. 100 significantly differentially expressed genes, expression of a Bowman-Birk trypsin inhibitor (BBI) gene was at least ca. 8-fold higher in Oh43 than in either parent. The Oh43 BBI gene was expressed as a recombinant protein. Purified BBI inhibited trypsin and the growth of fall armyworm larvae when added to insect diet. These experiments indicate that comparative gene expression analysis combined with insect resistance measurements of early inbreds can identify previously unrecognized resistance genes.

Expression of a wolf spider toxin in tobacco inhibits the growth of microbes and insects

Lycotoxin I, from the wolf spider (Lycosa carolinensis), is an amphipathic pore-forming peptide that has antimicrobial and anti-insect activity. Constitutive expression of a lycotoxin I modified for oral toxicity to insects in tobacco (Nicotiana tabacum) conferred significantly enhanced resistance to larvae of the corn earworm (Helicoverpa zea) and cigarette beetle (Lasioderma serricorne). Gene expression levels of modified lycotoxin I were negatively correlated to the survival of corn earworm larvae. In addition, pathogenic symptoms caused by Pseudomonas syringae pathovar tabaci and Alternaria alternata on the modified lycotoxin I-expressing leaves were significantly less severe than on wild type leaves. These results indicate that modified lycotoxin I expression in tobacco can potentially protect leaf tissue from a broad spectrum of pests and pathogens.

Corn Earworm Larvae (&lt;i&gt;Helicoverpa zea&lt;/i&gt;) Bounded on Warmer Direction (Thermotropism)

Thermotropism of corn earworm neonates was observed during Bt ( Bacillus thuringiensis ) toxicity bioassay against corn earworm ( Helicoverpa zea ). Six photos provided in this report convinced us that the corn earworm larvae moved to a warmer area within 24 hours of hatching. Possible reasons for the thermotropism are discussed but are not yet clear.

Expression of flavonoid 3’-hydroxylase is controlled by P1, the regulator of 3-deoxyflavonoid biosynthesis in maize

BackgroundThe maize (Zea mays) red aleurone1 (pr1) encodes a CYP450-dependent flavonoid 3’-hydroxylase (ZmF3’H1) required for the biosynthesis of purple and red anthocyanin pigments. We previously showed that Zmf3’h1 is regulated by C1 (Colorless1) and R1 (Red1) transcription factors. The current study demonstrates that, in addition to its role in anthocyanin biosynthesis, the Zmf3’h1 gene also participates in the biosynthesis of 3-deoxyflavonoids and phlobaphenes that accumulate in maize pericarps, cob glumes, and silks. Biosynthesis of 3-deoxyflavonoids is regulated by P1 (Pericarp color1) and is independent from the action of C1 and R1 transcription factors.ResultsIn maize, apiforol and luteoforol are the precursors of condensed phlobaphenes. Maize lines with functional alleles of pr1 and p1 (Pr1;P1) accumulate luteoforol, while null pr1 lines with a functional or non-functional p1 allele (pr1;P1 or pr1;p1) accumulate apiforol. Apiforol lacks a hydroxyl group at the 3’-position of the flavylium B-ring, while luteoforol has this hydroxyl group. Our biochemical analysis of accumulated compounds in different pr1 genotypes showed that the pr1 encoded ZmF3’H1 has a role in the conversion of mono-hydroxylated to bi-hydroxylated compounds in the B-ring. Steady state RNA analyses demonstrated that Zmf3’h1 mRNA accumulation requires a functional p1 allele. Using a combination of EMSA and ChIP experiments, we established that the Zmf3’h1 gene is a direct target of P1. Highlighting the significance of the Zmf3’h1 gene for resistance against biotic stress, we also show here that the p1 controlled 3-deoxyanthocyanidin and C-glycosyl flavone (maysin) defence compounds accumulate at significantly higher levels in Pr1 silks as compared to pr1 silks. By virtue of increased maysin synthesis in Pr1 plants, corn ear worm larvae fed on Pr1; P1 silks showed slower growth as compared to pr1; P1 silks.ConclusionsOur results show that the Zmf3’h1 gene participates in the biosynthesis of phlobaphenes and agronomically important 3-deoxyflavonoid compounds under the regulatory control of P1.