Diabrotica Research Articles

Field-Based Assessment of Resistance to Bt Corn by Western Corn Rootworm (Coleoptera: Chrysomelidae).

Western corn rootworm, Diabrotica virgifera virgifera LeConte, is a serious pest of corn and is managed with corn that produces insecticidal toxins derived from the bacterium Bacillus thuringiensis (Bt). Beginning in 2009, resistance to Cry3Bb1 corn, and severe injury to Cry3Bb1 corn in the field, was observed in Iowa. However, few data exist on how Cry3Bb1-resistant western corn rootworm interact with various management practices in the field. Using a field experiment, we measured adult emergence and feeding injury to corn roots for both Cry3Bb1-resistant and Cry3Bb1-susceptible populations of western corn rootworm when tested against various Bt corn hybrids and a soil-applied insecticide. Between 2012 and 2013, we evaluated five fields that were associated with greater than one node of feeding injury to Cry3Bb1 corn by western corn rootworm (i.e., problem-field populations), and a laboratory strain that had never been exposed to Bt corn (i.e., control population). Adult emergence for western corn rootworm and root injury to corn were significantly higher in problem-field populations than control populations for both Cry3Bb1 corn and mCry3A corn. By contrast, corn with Cry34/35Ab1, either alone or pyramided with Cry3Bb1, significantly reduced adult emergence and root injury in both problem fields and control fields. In problem fields, application of the soil-applied insecticide to Cry3Bb1 corn significantly reduced root injury, but not adult emergence. Our results are discussed in terms of developing strategies for managing western corn rootworm with resistance to Cry3Bb1 and mCry3A, and delaying the additional evolution of Bt resistance by this pest.

Multiple Assays Indicate Varying Levels of Cross Resistance in Cry3Bb1-Selected Field Populations of the Western Corn Rootworm to mCry3A, eCry3.1Ab, and Cry34/35Ab1.

Minnesota populations of Diabrotica virgifera virgifera LeConte, the western corn rootworm, surviving Cry3Bb1-expressing corn in the field and western corn rootworm populations assumed to be susceptible to all Bt proteins were evaluated for susceptibility to Cry3Bb1, mCry3A, eCry3.1Ab, and Cry34/35Ab1 in diet assays and three different plant-based assays. Rootworm populations originating from Cry3Bb1 fields and that consistently experienced greater than expected damage had increased survival and larval growth compared to control populations assayed on Cry3Bb1 as well as mCry3a and eCry3.1Ab. Cross resistance was documented between Cry3Bb1 and both mCry3A and eCry3.1Ab as single toxins. Despite very high resistance ratios in some comparisons, cross resistance was not complete and also varied with the population being evaluated, the trait measured, and the susceptible rootworm population used for comparison. Regardless of resistance and cross resistance, all proteins, even Cry3Bb1, retained some efficacy in terms of either reducing rootworm larval growth, protecting plants from damage, or both, for all rootworm populations evaluated. For one Cry3Bb1-selected population, a resistance ratio of 9.1-fold was found to Cry34/35Ab1 when evaluating EC 50 values relative to a susceptible control population; however, resistance to Cry34/35Ab1 was not evident in all assays in this population. The United States Environmental Protection Agency recently suggested eliminating diet assays as part of the Bt resistance monitoring process. However, given the variability of responses of western corn rootworm populations to different proteins in different assays, both plant and diet assays are needed as options for detecting and fully characterizing resistance.

IPM Use With the Deployment of a Non-High Dose Bt Pyramid and Mitigation of Resistance for Western Corn Rootworm (Diabrotica virgifera virgifera).

Recent detection of western corn rootworm resistance to Bt (Bacillus thuringiensis) corn prompted recommendations for the use of integrated pest management (IPM) with planting refuges to prolong the durability of Bt technologies. We conducted a simulation experiment exploring the effectiveness of various IPM tools at extending durability of pyramided Bt traits. Results indicate that some IPM practices have greater merits than others. Crop rotation was the most effective strategy, followed by increasing the non-Bt refuge size from 5 to 20%. Soil-applied insecticide use for Bt corn did not increase the durability compared with planting Bt with refuges alone, and both projected lower durabilities. When IPM participation with randomly selected management tools was increased at the time of Bt commercialization, durability of pyramided traits increased as well. When non-corn rootworm expressing corn was incorporated as an IPM option, the durability further increased. For corn rootworm, a local resistance phenomenon appeared immediately surrounding the resistant field (hotspot) and spread throughout the local neighborhood in six generations in absence of mitigation. Hotspot mitigation with random selection of strategies was ineffective at slowing resistance, unless crop rotation occurred immediately; regional mitigation was superior to random mitigation in the hotspot and reduced observed resistance allele frequencies in the neighborhood. As resistance alleles of mobile pests can escape hotspots, the scope of mitigation should extend beyond resistant sites. In the case of widespread resistance, regional mitigation was less effective at prolonging the life of the pyramid than IPM with Bt deployment at the time of commercialization.

A socioeconomic analysis of biocontrol in integrated pest management: A review of the effects of uncertainty, irreversibility and flexibility

European regulations on the sustainable use of pesticides aim to promote integrated pest management (IPM) strategy and the use of biological control agents. However, uncertainty over benefits and costs, irreversibility effects as well as flexibility in adoption of this technology needs to be considered. Economic evaluation of IPM using simple cost-benefit analysis may be inadequate. Therefore, the need to develop evaluation tools that takes the aforementioned constraints into consideration is imperative. To this end, we introduce the maximum incremental social tolerable irreversible costs (MISTICs) as a tool for such evaluation. Only when the incremental reversible benefits of the IPM strategy outweigh possible irreversible costs of such a strategy by a minimum threshold (hurdle rate) should introduction of biocontrol be considered. Our approach allows assessment of an IPM strategy from a private (farmers) point of view to be extended to a social context.We aim to evaluate the MISTICs value of biocontrol adoption for Western Corn Rootworm in maize cultivation and wireworms in potato cultivation for selected European Union member states (Germany, France, Austria, Spain and Italy) based on the option and net present value using the McDonalds-Siegel Stimulation. We also evaluate the potential irreversible costs associated with biocontrol introduction.The farm-level MISTICs per hectare for biocontrol of Western Corn Rootworm in maize cultivation in the selected countries were estimated to be less than €150/ha while biocontrol of wireworms in potato cultivation, well above €200/ha. These results suggest that the introduction of biocontrol for wireworms in potato cultivation, given the higher MISTICs values, is more economically viable compared to the biocontrol of Western Corn Rootworm in maize. The model can also be extended to include other factors such as regulatory hurdles.

MCry3A-Selected Western Corn Rootworm (Coleoptera: Chrysomelidae) Colony Exhibits High Resistance and Has Reduced Binding of mCry3A to Midgut Tissue.

Several Bt maize events expressing various insecticidal Cry protein genes have been commercialized for management of western corn rootworm, Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae). We used high efficacy (>99.7%) experimental maize events that express mCry3A for selections under laboratory conditions to develop a western corn rootworm colony resistant to mCry3A at higher levels than published results. The resistance ratio (RR) to mCry3A was >97-fold based on LC 50 values in diet-based bioassays after six generations of selections when compared to that of an unselected Control colony. Using a sublethal seedling assay (SSA) method, we confirmed that the colony had no cross-resistance to maize event DAS-59122-7, which expresses Cry34/35Ab. Reciprocal crosses between the mCry3A-resistant colony and the susceptible colony were performed to test the inheritance of resistance. Larval survival and development evaluated by the SSA method indicated that resistance to mCry3A was inherited autosomally and was incompletely recessive (h = 0.23-0.25). Specific binding of mCry3A to brush border membrane vesicles of midgut tissue revealed reduced binding in the resistant colony when compared to binding in the susceptible colony. This is the first report where resistance in western corn rootworm has been shown to involve reduced binding of a Cry3-class protein in midgut tissue.

Development of a CO2 -releasing coformulation based on starch, Saccharomyces cerevisiae and Beauveria bassiana attractive towards western corn rootworm larvae.

CO2 is known as an attractant for many soil-dwelling pests. To implement an attract-and-kill strategy for soil pest control, CO2 -emitting formulations need to be developed. The aim of the present work was to develop a slow-release bead system in order to bridge the gap between application and hatching of western corn rootworm larvae. We compared different Ca-alginate beads containing Saccharomyces cerevisiae for their potential to release CO2 over a period of several weeks. The addition of starch improved CO2 release, resulting in significantly higher CO2 concentrations in soil for at least 4 weeks. The missing amylase activity was compensated for either by microorganisms present in the soil or by coencapsulation of Beauveria bassiana. Formulations containing S. cerevisiae, starch and B. bassiana were attractive for western corn rootworm larvae within the first 4 h following exposure; however, when considering the whole testing period, the maize root systems remained more attractive for the larvae. Coencapsulation of S. cerevisiae, starch and B. bassiana is a promising approach for the development of attractive formulations for soil applications. For biological control strategies, the attractiveness needs to be increased by phagostimuli to extend contact between larvae and the entomopathogenic fungus growing out of these formulations. © 2016 Society of Chemical Industry.

The Effect of Western Corn Rootworm (Coleoptera: Chrysomelidae) and Water Deficit on Maize Performance Under Controlled Conditions.

The western corn rootworm, Diabrotica virgifera virgifera LeConte, is the most important insect of maize, Zea mays L., but knowledge of its interaction with water deficit on maize production is lacking. A series of greenhouse experiments using three infestation levels of the western corn rootworm, D. virgifera virgifera, under well-watered, moderately dry, and very dry soil moisture levels were conducted to quantify the interaction of western corn rootworm and soil water deficit on B73×Mo17 maize growth and physiology. Three separate experiments were conducted. Soil moisture regimes were initiated 30 d postplanting for experiments using neonate and second-instar larvae and 30 d postinfestation in the experiment using eggs. In the neonate and second-instar experiments, there were no significant differences among western corn rootworm levels in their effects on leaf water potential, shoot dry weight, and root dry weight. The interaction of western corn rootworm and soil moisture significantly impacted the larval recovery in the neonate experiment, but no other significant interactions were documented between soil moisture levels and rootworm infestation levels. Overall, the results indicate that under the conditions of these experiments, the effect of water deficit was much greater on plants than the effect of western corn rootworm and that the interactions between water deficit and western corn rootworm levels minimally affected the measured parameters of plant performance.

Influence of Planting Pattern on Pest Management in Virginia Market Type Peanut (Arachis hypogaea L.)

ABSTRACT Twelve field trials were completed in North Carolina from 2005 to 2007 to determine the impact of planting pattern on peanut yield, market grade characteristics, and pest reaction for the Virginia market type cultivars Perry and VA 98R. The first set of experiments was designed to evaluate incidence of Cylindrocladium black rot (caused by Cylindrocladium parasiticum) using a factorial treatment arrangement of two cultivars, two levels of fumigation (none or metam sodium at 112 L/ha), and two levels of planting pattern (single rows spaced 91-cm apart or twin rows spaced 18 cm apart on 91-cm centers). Visible symptoms associated with Cylindrocladium black rot were not affected by planting pattern. However, in 1 of 3 trials pod scarring as a result of feeding by southern corn rootworm (Diabrotica undecimpunctata Howardi) was greater in twin rows compared with single rows. Damage from tobacco thrips (Frankliniella fusca) was also higher in the cultivar VA 98R planted in twin rows than in single rows or for the cultivar Perry. In the second set of experiments designed to evaluate incidence of Sclerotinia blight (caused by Sclerotinia minor), a factorial treatment arrangement consisting of two cultivars, two levels of boscalid (none or boscalid at 0.44 kg ai/ha), and two levels of planting pattern was implemented. Boscalid decreased Sclerotinia blight incidence at one of 4 locations, and increased pod yield. Yield of Perry was lower when grown in twin rows versus single rows, but yield of VA 98R was higher in twin rows. Peanut planted in twin rows yielded more than peanut planted in single rows regardless of cultivar, fumigation, or disease history in most trials. Results from these experiments suggest that planting pattern can have a minor impact on response to pests and that yield and market grades are often improved when peanut is planted in twin-row planting patterns compared with single rows.

Use of chromatin remodeling ATPases as RNAi targets for parental control of western corn rootworm (Diabrotica virgifera virgifera) and Neotropical brown stink bug (Euschistus heros)

RNA interference (RNAi) is a gene silencing mechanism that is present in animals and plants and is triggered by double stranded RNA (dsRNA) or small interfering RNA (siRNA), depending on the organism. In the western corn rootworm (WCR), Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae), RNAi can be achieved by feeding rootworms dsRNA added to artificial diet or plant tissues transformed to express dsRNA. The effect of RNAi depends on the targeted gene function and can range from an absence of phenotypic response to readily apparent responses, including lethality. Furthermore, RNAi can directly affect individuals that consume dsRNA or the effect may be transferred to the next generation. Our previous work described the potential use of genes involved in embryonic development as a parental RNAi technology for the control of WCR. In this study, we describe the use of chromatin-remodeling ATPases as target genes to achieve parental gene silencing in two insect pests, a coleopteran, WCR, and a hemipteran, the Neotropical brown stink bug, Euschistus heros Fabricius (Hemiptera: Pentatomidae). Our results show that dsRNA targeting chromatin-remodeling ATPase transcripts, brahma, mi-2, and iswi strongly reduced the fecundity of the exposed females in both insect species. Additionally, knockdown of chd1 reduced the fecundity of E. heros.

Development and Characterization of MIR604 Resistance in a Western Corn Rootworm Population (Coleoptera: Chrysomelidae).

mCry3A is one of only four proteins licensed for commercial use in Diabrotica control. Utilizing a colony of western corn rootworm, Diabrotica virgifera virgifera LeConte, selected for resistance to mCry3A, we evaluated how mCry3A resistance was inherited and whether fitness costs were associated with mCry3A resistance. Reciprocal crosses between a selected colony and a control colony were performed; resulting progeny along with parent colonies were evaluated in dose toxicity assays, greenhouse assays, and seedling assays. Dose toxicity assay results were inconclusive, as the highest dose of protein tested did not produce sufficient mortality for accurate LC50 calculation. In whole-plant greenhouse assays on mCry3A-expressing corn, larval relative survival of the selected female × control male reciprocal cross was similar to that of the selected colony, while that of the control female × selected male was intermediate between the mCry3-selected colony and the control colony. However, when adult relative survival in whole-plant greenhouse assays was examined, no significant difference between the reciprocal crosses and the two parent colonies was detected. Heritability calculations based on both larval (0.66) and adult (1.03) survival data indicate that resistance to mCry3A is not inherited in a recessive manner. The selected colony was removed from selection pressure and evaluated after three or eight generations of removal. At three generations of removal from selection, a slight decrease in larval relative survival was detected compared with the selected colony. At eight generations of removal from selection, larval relative survival was comparable with that of the selected colony.

Sequential sampling plan for assessing corn rootworm (Coleoptera: Chrysomelidae) larval injury to Bt maize

In-field product performance assessments are an essential component of corn rootworm (Diabrotica spp.; CRW) resistance management plans for transgenic maize (Zea mays L.) products expressing proteins derived from the bacterium Bacillus thuringiensis (Bt). The goal of a successful field sampling program is to accurately characterize in-field product performance while also minimizing resource demand, as collection of maize root samples to evaluate CRW injury can present resource challenges such as labor intensiveness, potential safety issues, and a limited time window available for sampling. A resource-efficient sequential sampling plan was developed that utilizes data-driven root injury threshold values derived from benchmark product performance data for both single and pyramided Bt maize traits for CRW control. This sequential sampling methodology incorporates unbiased sampling and controlled false positive and false negative error rates, enabling accurate assessment decisions to be made with efficient resource use. Our proposed approach enables systematic and effective classification of in-field Bt maize product performance, with applications to other CRW control technologies besides Bt maize products.

Carbon dioxide receptor genes and their expression profile in Diabrotica virgifera virgifera.

BackgroundDiabrotica virgifera virgifera, western corn rootworm, is one of the most devastating species in North America. D. v. virgifera neonates crawl through the soil to locate the roots on which they feed. Carbon dioxide (CO2) is one of the important volatile cues that attract D. v. virgifera larvae to roots. ResultsIn this study, we identified three putative D. v. virgifera gustatory receptor genes (Dvv_Gr1, Dvv_Gr2, and Dvv_Gr3). Phylogenetic analyses confirmed their orthologous relationships with known insect CO2 receptor genes from Drosophila, mosquitoes, and Tribolium. The phylogenetic reconstruction of insect CO2 receptor proteins and the gene expression profiles were analyzed. Quantitative analysis of gene expression indicated that the patterns of expression of these three candidate genes vary among larval tissues (i.e., head, integument, fat body, and midgut) and different development stages (i.e., egg, three larval stages, adult male and female).Conclusion The Dvv_Gr2 gene exhibited highest expression in heads and neonates, suggesting its importance in allowing neonate larvae to orient to its host plant. Similar expression patterns across tissues and developmental stages for Dvv_Gr1 and Dvv_Gr3 suggest a potentially different role. Findings from this study will allow further exploration of the functional role of specific CO2 receptor proteins in D. v. virgifera.

Effectiveness of Bt Maize against Corn Rootworm (Coleoptera: Chrysomelidae) and Species Composition in South Dakota Ten Years Following the Introduction of Transgenic Maize

Corn rootworms, Diabrotica spp. (Coleoptera: Chrysomelidae), are the most economically important pest of maize in the United States. The larvae consume maize roots, contributing to poor nutrient uptake, lodging of the plants, and severe yield loss. Transgenic maize expressing toxins from the bacterium Bacillus thuringiensis (Bt) have been employed to combat the pest. However, corn rootworm resistance to Bt maize has recently been reported and spurred research on the effectiveness of Bt toxins against corn rootworm across the Corn Belt. Thus, our objectives were twofold. First, we quantified the efficacy of Bt maize hybrids against corn rootworm in South Dakota. Second, we surveyed populations of Diabrotica species in areas where Bt maize had been grown since its commercialization in 2003. Maize hybrids expressing Cry3Bb1, Cry34/35Ab1, and mCry3A were evaluated at two locations with previous corn rootworm infestations or in fields that were in continuous maize for three or more consecutive years. We reported damage to maize expressing Cry3Bb1 and mCry3A toxins. We also observed significantly higher numbers of western corn rootworm than northern corn rootworm in our Diabrotica species surveys, suggesting that western corn rootworm are the dominant species in the region. This research has implications for maize production and sustainable corn rootworm management in South Dakota and the Northern Plains, and will advance our knowledge of the incidence of resistance to Bt toxins and long-term impact of Bt hybrids on species diversity of Diabrotica.

Evaluation of Foliar Applied Pyrethroid Insecticides for Control of Adult Corn Rootworm in Corn, 2015: Table 1

The efficacy of foliar-applied insecticides was evaluated against adult corn rootworm (CRW) populations in field corn near Clay Center, NE, during 2015. In continuous corn environments high populations of CRW adults feed on corn silks and may interfere with pollination, reducing grain yield. Information is needed on the efficacy of foliar insecticides against adult CRW in reproductive stage corn. Adult CRW populations consisted of >95% WCR and <5% SCR. Experimental design was an RCB with four replicates. Plot …

Evaluation of Near Isoline Corn Rootworm Traited and Refuge Corn Hybrids in Combination With Soil Insecticides at Planting for Larval Corn Rootworm Control, 2015: Table 1

Near isoline corn rootworm (CRW), traited and refuge corn hybrids in combination with soil insecticides were evaluated for effectiveness of larval CRW control near Clay Center, NE during 2015. Trial site was late-planted corn and pumpkins (insecticide free) during 2014. Experimental design was a RCB with four replicates. Plot size was 4 rows × ∼70 ft length with 30-inch row spacing. Soil type was a Crete silt loam. ‘NK N68B-3122’ (contains AgriSure 3122 insect and herbicide traits), ‘NK N68B-3000GT’ (contains AgriSure 3000GT insect and herbicide traits), and ‘NK N68B-GT’ (contains AgriSure …

Rootworm management: status of GM traits, insecticides and potential new tools.

Abstract Western corn rootworm ( Diabrotica virgifera virgifera LeConte) and northern corn rootworm ( Diabrotica barberi (Smith and Lawrence)) are major pests of maize in the USA. These pests have been managed with a variety of tactics over the last century. Both Diabrotica spp. have adapted to crop rotation in different ways in certain regions of the USA as well as to some of the insecticides targeted at them. D. v. virgifera has adapted to more of the chemical control measures and transgenic control methods. Discussed in this review are the challenges associated with managing both species, and how current management strategies might be combined and implemented to help manage damage from these species. Also, we discuss the potential for new technologies, such as RNA interference, to be used in the future.

Evaluation of Foliar-Applied Insecticides for Control of Adult Corn Rootworm in Corn, 2015C: Table 1

The efficacy of foliar applied insecticides was evaluated against adult corn rootworm (CRW) populations in field corn near Clay Center, NE during 2015. Adult CRW populations consisted of > 95% WCR and <5% SCR. Experimental design was a RCB with four replicates. Plot size was 8 rows × 30 ft length with 30-inch row spacing. ‘Pioneer P0157R’ corn hybrid was planted on 28 May …

Evaluation of Foliar Applied Insecticides for Control of Adult Corn Rootworm in Corn, 2015: Table 1

The efficacy of foliar applied insecticides was evaluated against adult corn rootworm (CRW) populations in field corn near Clay Center, NE during 2015. Adult CRW populations consisted of > 95% WCR and <5% SCR. Experimental design was a RCB with four replicates. Plot size was 8 rows × 30 ft length with 30-inch row spacing. ‘Pioneer P0157R’ corn hybrid was planted on 28 May …

Evaluation of Corn Rootworm Traited and Refuge Hybrids in Combination With Aztec 4.67G Insecticide at Planting for Control of Larval Corn Rootworm, 2015: Table 1

Single corn rootworm (CRW) traited and refuge hybrids in combination with Aztec 4.67G insecticide at planting were evaluated for effectiveness of larval CRW control near Clay Center, NE during 2015. The trial site was late-planted corn and pumpkins (insecticide free) during 2014. The experimental design was a RCB with four replicates. Plot size was 4 rows × ∼70 ft length with 30-inch row spacing. Soil type was a Crete silt loam. ‘DeKalb DKC62-95’ (contains Roundup Ready 2 herbicide trait [refuge hybrid]), ‘DeKalb DKC62-97RIB’ (contains 90% GENVT3PRIB traited seed and 10% refuge seed), ‘Pioneer P1395AM1’ (contains 90% Herculex XTRA traited seed and 10% Herculex I traited seed), and ‘NK N68B-3111’ …

Evaluation of Rootworm Management Options Using Different Corn Hybrids with Insecticides and Without, 2015.: Table 1

Corn rootworm (NCR/WCR) management efficacy trials were conducted to evaluate transgenic insect-resistant hybrids and granular insecticide combination treatments near Bryant, Cavour and Colman in South Dakota. The experimental plots were arranged in an RCBD with four ∼50 foot long replications. Four row treatment plots were established using rows that were spaced 30 inches apart at planting-time. Granular insecticides were applied with Noble metering units or the SmartBox delivery system. All metering units were mounted on a 4-row Kinze corn …