Corn Rootworm: Biology, Ecology, Behavior, and Integrated Management.

Species of the beetle genus Diabrotica (Coleoptera: Chrysomelidae) are native to North and South America, with their greatest diversity occurring in neotropical areas [...].

Movement Ecology of Adult Western Corn Rootworm: Implications for Management.

Movement of adult western corn rootworm, Diabrotica virgifera virgifera LeConte, is of fundamental importance to this species' population dynamics, ecology, evolution, and interactions with its environment, including cultivated cornfields. Realistic parameterization of dispersal components of models is needed to predict rates of range expansion, development, and spread of resistance to control measures and improve pest and resistance management strategies. However, a coherent understanding of western corn rootworm movement ecology has remained elusive because of conflicting evidence for both short- and long-distance lifetime dispersal, a type of dilemma observed in many species called Reid's paradox. Attempts to resolve this paradox using population genetic strategies to estimate rates of gene flow over space likewise imply greater dispersal distances than direct observations of short-range movement suggest, a dilemma called Slatkin's paradox. Based on the wide-array of available evidence, we present a conceptual model of adult western corn rootworm movement ecology under the premise it is a partially migratory species. We propose that rootworm populations consist of two behavioral phenotypes, resident and migrant. Both engage in local, appetitive flights, but only the migrant phenotype also makes non-appetitive migratory flights, resulting in observed patterns of bimodal dispersal distances and resolution of Reid's and Slatkin's paradoxes.

Soybean Foliage Consumption Reduces Adult Western Corn Rootworm (Diabrotica virgifera virgifera)(Coleoptera: Chrysomelidae) Survival and Stimulates Flight.

Western corn rootworm, Diabrotica virgifera virgifera LeConte, biology is tied to the continuous availability of its host (corn, Zea mays L.). Annual rotation of corn with a nonhost, like soybean (Glycine max (L.) Merrill) was a reliable tactic to manage western corn rootworm. Behavioral resistance to annual crop rotation (rotation resistance) allowed some eastern U.S. Corn Belt populations to circumvent rotation by laying eggs in soybean and in cornfields. When active in soybean, rotation-resistant adults commonly consume foliage, in spite of detrimental effects on beetle survival. Rotation-resistant beetle activity in soybean is enabled by the expression of certain proteinases and an adapted gut microbiota that provide limited protection from soybean antiherbivore defenses. We investigated the effects of corn and soybean herbivory on rotation-resistant female survival and initiation of flight using mortality assays and wind tunnel flight tests. Among field-collected females tested with mortality assays, beetles from collection sites in a cornfield survived longer than those from collection sites in a soybean field. However, reduced survival due to soybean herbivory could be restored by consuming corn tissues. Field-collected beetles that fed on a soybean tissue laboratory diet or only water were more likely to fly in a wind tunnel than corn-feeding beetles. Regardless of collection site and laboratory diet, 90.5% of beetles that flew oriented their flights upwind. Diet-related changes in the probability of flight provide a proximate mechanism for interfield movement that facilitates restorative feeding and the survival of females previously engaged in soybean herbivory.

Selinene Volatiles Are Essential Precursors for Maize Defense Promoting Fungal Pathogen Resistance.

To ensure food security, maize (Zea mays) is a model crop for understanding useful traits underlying stress resistance. In contrast to foliar biochemicals, root defenses limiting the spread of disease remain poorly described. To better understand belowground defenses in the field, we performed root metabolomic profiling and uncovered unexpectedly high levels of the sesquiterpene volatile ß-selinene and the corresponding nonvolatile antibiotic derivative ß-costic acid. The application of metabolite-based quantitative trait locus mapping using biparental populations, genome-wide association studies, and near-isogenic lines enabled the identification of terpene synthase21 (ZmTps21) on chromosome 9 as a ß-costic acid pathway candidate gene. Numerous closely examined ß-costic acid-deficient inbred lines were found to harbor Zmtps21 pseudogenes lacking conserved motifs required for farnesyl diphosphate cyclase activity. For biochemical validation, a full-length ZmTps21 was cloned, heterologously expressed in Escherichia coli, and demonstrated to cyclize farnesyl diphosphate, yielding ß-selinene as the dominant product. Consistent with microbial defense pathways, ZmTps21 transcripts strongly accumulate following fungal elicitation. Challenged field roots containing functional ZmTps21 alleles displayed ß-costic acid levels over 100 µg g-1 fresh weight, greatly exceeding in vitro concentrations required to inhibit the growth of five different fungal pathogens and rootworm larvae (Diabrotica balteata). In vivo disease resistance assays, using ZmTps21 and Zmtps21 near-isogenic lines, further support the endogenous antifungal role of selinene-derived metabolites. Involved in the biosynthesis of nonvolatile antibiotics, ZmTps21 exists as a useful gene for germplasm improvement programs targeting optimized biotic stress resistance.

Emergence and Abundance of Western Corn Rootworm (Coleoptera: Chrysomelidae) in Bt Cornfields With Structured and Seed Blend Refuges.

To slow evolution of western corn rootworm (Diabrotica virgifera virgifera LeConte) resistance to Bt (Bacillus thuringiensis Berliner) corn hybrids, non-Bt "refuges" must be planted within or adjacent to Bt cornfields, allowing susceptible insects to develop without exposure to Bt toxins. Bt-susceptible adults from refuges are expected to find and mate with resistant adults that have emerged from Bt corn, reducing the likelihood that Bt-resistant offspring are produced. The spatial and temporal distribution of adults in four refuge treatments (20, 5, and 0% structured refuges and 5% seed blend) and adjacent soybean fields was compared from 2010 to 2012. Adult emergence (adults/trap/day) from refuge corn in structured refuge treatments was greater than that from Bt corn, except during the post-pollination period of corn phenology when emergence from refuge and Bt plants was often the same. Abundance of free-moving adults was greatest in and near refuge rows in structured refuge treatments during vegetative and pollination periods. By post-pollination, adult abundance became evenly distributed. In contrast, adult abundance in 5% seed blends and 0% refuges was evenly distributed, or nearly so, across plots throughout the season. The persistent concentration of adults in refuge rows suggests that structured refuge configurations may not facilitate the expected mixing of adults from refuge and Bt corn. Seed blends produce uniform distributions of adults across the field that may facilitate mating between Bt and refuge adults and ultimately delay the evolution of Bt resistance.

Patterns of differential gene expression in adult rotation-resistant and wild-type western corn rootworm digestive tracts.

The western corn rootworm (WCR,Diabrotica virgifera virgifera LeConte) is an important pest of corn. Annual crop rotation between corn and soybean disrupts the corn-dependent WCR life cycle and is widely adopted to manage this pest. This strategy selected for rotation-resistant (RR) WCR with reduced ovipositional fidelity to corn. Previous studies revealed that RR-WCR adults exhibit greater tolerance of soybean diets, different gut physiology, and host-microbe interactions compared to rotation-susceptible wild types (WT). To identify the genetic mechanisms underlying these phenotypic changes, a de novo assembly of the WCR adult gut transcriptome was constructed and used for RNA-sequencing analyses of RNA libraries from different WCR phenotypes fed with corn or soybean diets. Global gene expression profiles of WT- and RR-WCR were similar when feeding on corn diets, but different when feeding on soybean. Using network-based methods, we identified gene modules transcriptionally correlated with the RR phenotype. Gene ontology enrichment analyses indicated that the functions of these modules were related to metabolic processes, immune responses, biological adhesion, and other functions/processes that appear to correlate to documented traits in RR populations. These results suggest that gut transcriptomic divergence correlated with brief soybean feeding and other physiological traits may exist between RR- and WT-WCR adults.

Differential effects of RNAi treatments on field populations of the western corn rootworm.

RNA interference (RNAi) mediated crop protection against insect pests is a technology that is greatly anticipated by the academic and industrial pest control communities. Prior to commercialization, factors influencing the potential for evolution of insect resistance to RNAi should be evaluated. While mutations in genes encoding the RNAi machinery or the sequences targeted for interference may serve as a prominent mechanism of resistance evolution, differential effects of RNAi on target pests may also facilitate such evolution. However, to date, little is known about how variation of field insect populations could influence the effectiveness of RNAi treatments. To approach this question, we evaluated the effects of RNAi treatments on adults of three western corn rootworm (WCR; Diabrotica virgifera virgifera LeConte) populations exhibiting different levels of gut cysteine protease activity, tolerance of soybean herbivory, and immune gene expression; two populations were collected from crop rotation-resistant (RR) problem areas and one from a location where RR was not observed (wild type; WT). Our results demonstrated that RNAi targeting DvRS5 (a highly expressed cysteine protease gene) reduced gut cysteine protease activity in all three WCR populations. However, the proportion of the cysteine protease activity that was inhibited varied across populations. When WCR adults were treated with double-stranded RNA of an immune gene att1, different changes in survival among WT and RR populations on soybean diets occurred. Notably, for both genes, the sequences targeted for RNAi were the same across all populations examined. These findings indicate that the effectiveness of RNAi treatments could vary among field populations depending on their physiological and genetic backgrounds and that the consistency of an RNAi trait's effectiveness on phenotypically different populations should be considered or tested prior to wide deployment. Also, genes that are potentially subjected to differential selection in the field should be avoided for RNAi-based pest control.

Modeling a western corn rootworm, Diabrotica virgifera virgifera (Coleoptera: Chrysomelidae), maturation delay and resistance evolution in Bt corn.

BACKGROUND: Emergence delay and female-skewed sex ratios among adults of Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae) from Bt corn have been reported in field studies. The authors used a simulation model to study the effect of a maturation delay and a female-skewed sex ratio for D. v. virgifera emerging from Bt corn on the evolution of Bt resistance. RESULTS: The effect of skewed toxin mortality in one sex on evolution of Bt resistance was insignificant. An emergence delay among resistant beetles from Bt corn slowed resistance evolution. A shift in the time of emergence for homozygous susceptible beetles from Bt corn did not have a significant effect on the evolution of Bt resistance in D. v. virgifera. CONCLUSION: This simulation study suggested that skewed toxin mortality in one sex and an emergence delay for beetles in Bt corn are not major concerns for managing resistance by D. v. virgifera to single-toxin or pyramided Bt corn.

Gut bacteria facilitate adaptation to crop rotation in the western corn rootworm.

Insects are constantly adapting to human-driven landscape changes; however, the roles of their gut microbiota in these processes remain largely unknown. The western corn rootworm (WCR, Diabrotica virgifera virgifera LeConte) (Coleoptera: Chrysomelidae) is a major corn pest that has been controlled via annual rotation between corn (Zea mays) and nonhost soybean (Glycine max) in the United States. This practice selected for a "rotation-resistant" variant (RR-WCR) with reduced ovipositional fidelity to cornfields. When in soybean fields, RR-WCRs also exhibit an elevated tolerance of antiherbivory defenses (i.e., cysteine protease inhibitors) expressed in soybean foliage. Here we show that gut bacterial microbiota is an important factor facilitating this corn specialist's (WCR's) physiological adaptation to brief soybean herbivory. Comparisons of gut microbiota between RR- and wild-type WCR (WT-WCR) revealed concomitant shifts in bacterial community structure with host adaptation to soybean diets. Antibiotic suppression of gut bacteria significantly reduced RR-WCR tolerance of soybean herbivory to the level of WT-WCR, whereas WT-WCR were unaffected. Our findings demonstrate that gut bacteria help to facilitate rapid adaptation of insects in managed ecosystems.

Abnormally high digestive enzyme activity and gene expression explain the contemporary evolution of a Diabrotica biotype able to feed on soybeans.

Western corn rootworm (Diabrotica virgifera) (WCR) depends on the continuous availability of corn. Broad adoption of annual crop rotation between corn (Zea mays) and nonhost soybean (Glycine max) exploited WCR biology to provide excellent WCR control, but this practice dramatically reduced landscape heterogeneity in East-central Illinois and imposed intense selection pressure. This selection resulted in behavioral changes and "rotation-resistant" (RR) WCR adults. Although soybeans are well defended against Coleopteran insects by cysteine protease inhibitors, RR-WCR feed on soybean foliage and remain long enough to deposit eggs that will hatch the following spring and larvae will feed on roots of planted corn. Other than documenting changes in insect mobility and egg laying behavior, 15 years of research have failed to identify any diagnostic differences between wild-type (WT)- and RR-WCR or a mechanism that allows for prolonged RR-WCR feeding and survival in soybean fields. We documented differences in behavior, physiology, digestive protease activity (threefold to fourfold increases), and protease gene expression in the gut of RR-WCR adults. Our data suggest that higher constitutive activity levels of cathepsin L are part of the mechanism that enables populations of WCR to circumvent soybean defenses, and thus, crop rotation. These new insights into the mechanism of WCR tolerance of soybean herbivory transcend the issue of RR-WCR diagnostics and management to link changes in insect gut proteolytic activity and behavior with landscape heterogeneity. The RR-WCR illustrates how agro-ecological factors can affect the evolution of insects in human-altered ecosystems.

Seeds of change: corn seed mixtures for resistance management and integrated pest management.

The use of mixtures of transgenic insecticidal seed and nontransgenic seed to provide an in-field refuge for susceptible insects in insect-resistance-management (IRM) plans has been considered for at least two decades. However, the U.S. Environmental Protection Agency has only recently authorized the practice. This commentary explores issues that regulators, industry, and other stakeholders should consider as the use of biotechnology increases and seed mixtures are implemented as a major tactic for IRM. We discuss how block refuges and seed mixtures in transgenic insecticidal corn, Zea mays L., production will influence integrated pest management (IPM) and the evolution of pest resistance. We conclude that seed mixtures will make pest monitoring more difficult and that seed mixtures may make IRM riskier because of larval behavior and greater adoption of insecticidal corn. Conversely, block refuges present a different suite of risks because of adult pest behavior and the lower compliance with IRM rules expected from farmers. It is likely that secondary pests not targeted by the insecticidal corn as well as natural enemies will respond differently to block refuges and seed mixtures.

Refuge or reservoir? The potential impacts of the biofuel crop Miscanthus x giganteus on a major pest of maize.

BACKGROUND: Interest in the cultivation of biomass crops like the C4 grass Miscanthus x giganteus (Miscanthus) is increasing as global demand for biofuel grows. In the US, Miscanthus is promoted as a crop well-suited to the Corn Belt where it could be cultivated on marginal land interposed with maize and soybean. Interactions (direct and indirect) of Miscanthus, maize, and the major Corn Belt pest of maize, the western corn rootworm, (Diabrotica virgifera virgifera LeConte, WCR) are unknown. Adding a perennial grass/biomass crop to this system is concerning since WCR is adapted to the continuous availability of its grass host, maize (Zea mays). METHODOLOGY/PRINCIPAL FINDINGS: In a greenhouse and field study, we investigated WCR development and oviposition on Miscanthus. The suitability of Miscanthus for WCR development varied across different WCR populations. Data trends indicate that WCR populations that express behavioural resistance to crop rotation performed as well on Miscanthus as on maize. Over the entire study, total adult WCR emergence from Miscanthus (212 WCR) was 29.6% of that from maize (717 WCR). Adult dry weight was 75-80% that of WCR from maize; female emergence patterns on Miscanthus were similar to females developing on maize. There was no difference in the mean no. of WCR eggs laid at the base of Miscanthus and maize in the field. CONCLUSIONS/SIGNIFICANCE: Field oviposition and significant WCR emergence from Miscanthus raises many questions about the nature of likely interactions between Miscanthus, maize and WCR and the potential for Miscanthus to act as a refuge or reservoir for Corn Belt WCR. Responsible consideration of the benefits and risks associated with Corn Belt Miscanthus are critical to protecting an agroecosystem that we depend on for food, feed, and increasingly, fuel. Implications for European agroecosystems in which Miscanthus is being proposed are also discussed in light of the WCR's recent invasion into Europe.

Use of transgenic plants to measure insect herbivore movement.

Use of ingested transgenic corn tissue as a marker for measuring movement of adult Diabrotica virgifera virgifera (LeConte) (Coleoptera: Chrysomelidae; western corn rootworm) was investigated. Laboratory observations of beetles feeding on corn foliage, pollen, silks, or soybean foliage provided background on feeding patterns. The interval between food consumption and its appearance in feces (gut passage time) ranged from 102.7 +/- 11 min for soybean foliage to 56.7 +/- 2.9 min for corn silks. In a laboratory assay, protein expression tests identified the presence of Cry3Bb1 protein inside 50% of adult D. virgifera for up to 16 h after they had last consumed Cry3Bb1 protein-expressing corn silks from 'YieldGard Rootworm' corn plants (Monsanto Co.). Cry3Bb1 protein could not be detected by 32 h postfeeding. The proportion of Cry3Bb1 protein-positive beetles declined linearly with increasing time since feeding on 'YieldGard Rootworm' tissue. Approximately 20% of adult D. virgifera collected near 'YieldGard Rootworm' corn plots tested positive for Cry3Bb1 protein, indicating 'YieldGard Rootworm' tissue consumption within the last 16-32 h. Based on a 16- to 32-h postfeeding detection interval for Cry3Bb1 protein and the distance between 'YieldGard Rootworm' sources and sites where Cry3Bb1-positive insects were collected, 85.3% of males and females moved < or = 4.6-9.1 m/d through R2-R3 stage corn. Among Cry3Bb1-positive adults that left corn and were captured in an adjacent soybean field, 86.4% of males and 93.1% of females moved < or = 4.6-9.1 m/d through soybean. Detection of transgenic plant tissues in mobile insect herbivores is a novel application of biotechnology to the study of insect movement.