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 [...].

Resistance management and integrated pest management insights from deployment of a Cry3Bb1+ Gpp34Ab1/Tpp35Ab1 pyramid in a resistant western corn rootworm landscape.

In Nebraska USA, many populations of western corn rootworm (WCR), Diabrotica virgifera virgifera LeConte, now exhibit some level of resistance to all corn rootworm-active Bacillus thuringiensis Berliner (Bt) proteins expressed in commercial hybrids. Therefore, a study was conducted in northeast Nebraska from 2020-2022 to reevaluate current corn rootworm management options in continuous maize (consecutive planting for ≥2 years). Results from on-farm experiments to evaluate a standard soil-applied insecticide (Aztec® 4.67G) in combination with non-rootworm Bt or rootworm-active Bt pyramided maize (Cry3Bb1 + Gpp34Ab1/Tpp35Ab1) are reported within the context of WCR Bt resistance levels present. Corrected survival from Bt pyramid single-plant bioassays (<0.3, 0.3-0.49, >0.5) was used to place populations into 3 resistance categories. Variables evaluated included root injury, adult emergence, proportion lodged maize, and grain yield. Key results: A composite analysis of all populations across resistance levels indicated that addition of soil insecticide to Bt pyramid significantly reduced adult emergence and lodging but did not significantly increase root protection or yield. Within and among resistance category analyses of root injury revealed that the Bt pyramid remained highly efficacious at any non-rootworm Bt root injury level when resistance was absent or low. When corrected survival was >0.3, mean Bt pyramid root injury tracked more closely in a positive linear fashion with mean non-rootworm Bt root injury (rootworm density x level of resistance interaction). Similar trends were obtained for adult emergence but not yield. Mean Bt pyramid root injury rating was <0.75 in most populations with Bt resistance, which contributed to no significant yield differences among categories. Results are discussed within the context of IPM:IRM tradeoffs and the need to reduce WCR densities in this system to decrease the impact of the density x resistance interaction to bridge use of current pyramids with new technologies introduced over the next decade.

Susceptibility of northern corn rootworm (Diabrotica barberi) populations to Cry3Bb1 and Gpp34/Tpp35Ab1 proteins in seedling and diet overlay toxicity assays.

The northern corn rootworm, Diabrotica barberi Smith & Lawrence (Coleoptera: Chrysomelidae) is a major pest of maize in the United States Corn Belt. Recently, resistance to Bacillus thuringiensis (Bt) maize was reported in North Dakota and increased use of Bt maize hybrids could facilitate resistance evolution in other maize-producing states. In this study, susceptibility to Bt proteins was evaluated in wild D. barberi populations from 8 fields collected in 5 different states (Minnesota, Missouri, Nebraska, Iowa, and North Dakota). Field populations were compared to a susceptible D. barberi colony in seedling and diet toxicity assays conducted with 3 concentrations of Cry3Bb1 (0.4, 4.0, and 40.0 µg/cm2) and Gpp34/Tpp35Ab1 (previously called Cry34/35Ab1; 1.4, 14.0, and 140.0 µg/cm2). The 2019 population from Meeker Co., Minnesota (MN-2019), exhibited the lowest mortality to Cry3Bb1 and also had nominally lowest mortality to Gpp34/Tpp35Ab1 at the highest concentrations tested in diet toxicity assays. Percent second instar was also highest for larvae of the Minnesota population surviving Cry3Bb1. In seedling assays, MN and IA-2018 populations exhibited the highest proportion survival and dry weight to both proteins expressed in corn. No significant differences in mortality, percent second instar, and dry weight were observed at the highest concentration for both proteins among the populations collected in in 2020. Most D. barberi populations were still highly susceptible to Cry3Bb1 and Gpp34/Tpp35Ab1 proteins based on diet and seedling assays, but resistance appears to be developing in some D. barberi populations. Now that methods are available, resistance monitoring may also be needed for D. barberi in some regions.

Effects of SmartStax® and SmartStax® PRO maize on western corn rootworm (Diabrotica virgifera virgifera LeConte) larval feeding injury and adult life history parameters.

Field-evolved resistance of the western corn rootworm (WCR), Diabrotica virgifera virgifera LeConte, to Bacillus thuringiensis Berliner (Bt) proteins Cry3Bb1 and Cry34/35Ab1 (now classified as Gpp34Ab1/Tpp35Ab1) expressed in the pyramid SmartStax® has been documented in areas of the United States (U.S.) Corn Belt. SmartStax® PRO is a recently registered rootworm-active pyramid containing the same Bt proteins expressed in SmartStax® plus DvSnf7 dsRNA. Little to no published data is available comparing efficacy of the technologies or potential effects of dietary exposure on adult WCR fitness. Therefore, experiments were conducted to compare effects of adult WCR dietary exposure to SmartStax® and SmartStax® PRO on life history parameters and efficacy of the technologies in the field with both Bt-susceptible and Bt-resistant WCR populations. WCR life history parameters evaluated included adult longevity, head capsule width, egg production, and egg viability. Results of small-plot field trials indicated that both technologies provided a high level of root protection when a Bt-susceptible WCR population was present. Root protection was reduced on SmartStax® but maintained on SmartStax® PRO when WCR Bt resistance occurred. Lifetime egg production was the key life history parameter that was significantly reduced when either Bt-susceptible or Bt-resistant adult WCR were fed SmartStax® or SmartStax® PRO diet. A potential fitness advantage was apparent as egg production was significantly higher in the Bt-resistant than Bt-susceptible population. The similar response by the Bt-susceptible WCR population to SmartStax® and SmartStax® PRO indicates that results were caused by sublethal dietary exposure to Bt proteins. Adult size (males < females) and egg viability (high: >95%) were not significantly different among treatments but longevity results were inconsistent between years. Collectively, the field efficacy and life history parameter data expand existing knowledge of SmartStax® and SmartStax® PRO technologies, which will inform practical WCR resistance management programs.

Characterization of western corn rootworm (Coleoptera: Chrysomelidae) susceptibility to foliar insecticides in northeast Nebraska.

Foliar-applied insecticides are commonly used for adult western corn rootworm (WCR), Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae), control in Nebraska but little efficacy data is available. Anecdotal reports of reduced efficacy in areas of northeast Nebraska led to the conduct of this study. Objectives were to (i) evaluate the efficacy of commercial applications of commonly used formulated insecticides (bifenthrin, lambda-cyhalothrin, chlorpyrifos, or tank mixes) for WCR control in 7 northeast Nebraska counties during 2019 and 2020 and (ii) conduct adult WCR concentration-response vial bioassays with bifenthrin, chlorpyrifos, and dimethoate active ingredients on a subset of field populations. Whole plant counts (WPC) were used to measure WCR densities in insecticide-treated and untreated maize fields before and after insecticide application. Field control was excellent with organophosphate/pyrethroid tank mixes as proportional change in mean WPC of treated fields was significantly reduced (>0.90) versus untreated fields where little change in WPC occurred. The exception was one treated Boone County field where proportional reduction in WPC was ≤0.78. Bioassays revealed LC50s and resistance ratios of most populations exposed to bifenthrin and dimethoate were not significantly different than the susceptible control. Most populations exhibited a low level of chlorpyrifos resistance when compared to the susceptible control. Field and lab data suggest the local onset of practical WCR field-evolved resistance to bifenthrin in Boone County and chlorpyrifos in Boone and Colfax counties. Results of this study will increase our understanding of WCR resistance evolution, serve as a comprehensive baseline for future research, and inform WCR management programs.

A draft Diabrotica virgifera virgifera genome: insights into control and host plant adaption by a major maize pest insect.

BACKGROUND: Adaptations by arthropod pests to host plant defenses of crops determine their impacts on agricultural production. The larval host range of western corn rootworm, Diabrotica virgifera virgifera (Coleoptera: Chrysomelidae), is restricted to maize and a few grasses. Resistance of D. v. virgifera to crop rotation practices and multiple insecticides contributes to its status as the most damaging pest of cultivated maize in North America and Europe. The extent to which adaptations by this pest contributes to host plant specialization remains unknown. RESULTS: A 2.42 Gb draft D. v. virgifera genome, Dvir_v2.0, was assembled from short shotgun reads and scaffolded using long-insert mate-pair, transcriptome and linked read data. K-mer analysis predicted a repeat content of ≥ 61.5%. Ortholog assignments for Dvir_2.0 RefSeq models predict a greater number of species-specific gene duplications, including expansions in ATP binding cassette transporter and chemosensory gene families, than in other Coleoptera. A majority of annotated D. v. virgifera cytochrome P450s belong to CYP4, 6, and 9 clades. A total of 5,404 transcripts were differentially-expressed between D. v. virgifera larvae fed maize roots compared to alternative host (Miscanthus), a marginal host (Panicum virgatum), a poor host (Sorghum bicolor) and starvation treatments; Among differentially-expressed transcripts, 1,908 were shared across treatments and the least number were between Miscanthus compared to maize. Differentially-expressed transcripts were enriched for putative spliceosome, proteosome, and intracellular transport functions. General stress pathway functions were unique and enriched among up-regulated transcripts in marginal host, poor host, and starvation responses compared to responses on primary (maize) and alternate hosts. CONCLUSIONS: Manual annotation of D. v. virgifera Dvir_2.0 RefSeq models predicted expansion of paralogs with gene families putatively involved in insecticide resistance and chemosensory perception. Our study also suggests that adaptations of D. v. virgifera larvae to feeding on an alternate host plant invoke fewer transcriptional changes compared to marginal or poor hosts. The shared up-regulation of stress response pathways between marginal host and poor host, and starvation treatments may reflect nutrient deprivation. This study provides insight into transcriptomic responses of larval feeding on different host plants and resources for genomic research on this economically significant pest of maize.

Reduced susceptibility of western corn rootworm (Diabrotica virgifera virgifera LeConte) populations to Cry34/35Ab1-expressing maize in northeast Nebraska.

The western corn rootworm (WCR; Diabrotica virgifera virgifera LeConte) is a significant pest of maize (Zea mays L.) across the United States Corn Belt. Transgenic maize hybrids expressing insecticidal proteins derived from Bacillus thuringiensis (Bt) have been used to manage WCR since 2003. Widespread resistance to Cry3Bb1 (and associated cross-resistance to mCry3A and eCry3.1Ab) has placed increased selection pressure on Cry34/35Ab1 in single-protein and pyramided transgenic maize hybrids. Data on the susceptibility of Nebraska WCR populations to Cry34/35Ab1 has not been published since 2015 and plant-based bioassays conducted in 2017-2018 confirmed resistance to Cry3Bb1 + Cry34/35Ab1 maize, suggesting resistance to Cry34/35Ab1 has evolved in the Nebraska landscape. Therefore, plant-based bioassays were conducted on F1 progeny of WCR populations collected from northeast Nebraska in 2018 and 2019. Larval survival and development were used to classify resistance to Cry34/35Ab1 in each WCR population. Bioassays confirmed incomplete resistance to Cry34/35Ab1 maize in 21 of 30 WCR populations; 9 of 30 WCR populations remained susceptible to Cry34/35Ab1. Collectively, results indicate that northeast Nebraska WCR populations were in the initial stages of resistance evolution to Cry34/35Ab1 during 2018-2019. Appropriate resistance management strategies are needed to mitigate resistance and preserve efficacy of rootworm-active products containing Cry34/35Ab1.

Characterizing the sublethal effects of SmartStax PRO dietary exposure on life history traits of the western corn rootworm, Diabrotica virgifera virgifera LeConte.

The western corn rootworm (WCR), Diabrotica virgifera virgifera LeConte, is an economically important pest of field corn (Zea mays L.) across the United States (U.S.) Corn Belt. Repeated use of transgenic hybrids expressing Bacillus thuringiensis (Bt) proteins has selected for field-evolved resistance to all current rootworm-active Bt proteins. The newest product available for WCR management is SmartStax® PRO, a rootworm-active pyramid containing Cry3Bb1, Cry34/35Ab1 [now reclassified as Gpp34Ab1/Tpp35Ab1] and a new mode of action, DvSnf7 dsRNA. Understanding the fitness of adult WCR after dietary exposure to SmartStax® PRO will identify potential impacts on WCR population dynamics and inform efforts to optimize resistance management strategies. Therefore, the objective of the present study was to characterize the effect of SmartStax® PRO dietary exposure on WCR life history traits. Adult WCR were collected during 2018 and 2019 from emergence tents placed over replicated field plots of SmartStax® PRO or non-rootworm Bt corn at a site with a history of rootworm-Bt trait use and suspected resistance to Cry3Bb1 and Cry34/35Ab1. Adult survival was reduced by 97.1-99.7% in SmartStax® PRO plots relative to the non-rootworm Bt corn plots during the study. Individual male/female pairs were fed different diets of ear tissue to simulate lifetime or adult exposure. Life history parameters measured included adult longevity, adult head capsule width, lifetime female egg production, and egg viability. Results indicate that lifetime or adult exposure to SmartStax® PRO significantly reduced adult longevity and lifetime egg production. Larval exposure to SmartStax® PRO significantly reduced WCR adult size. Results from this study collectively suggest that SmartStax® PRO may negatively impact WCR life history traits, which may lead to reduced population growth when deployed in an area with WCR resistance to Bt traits.

Evidence of western corn rootworm (Diabrotica virgifera virgifera LeConte) field-evolved resistance to Cry3Bb1 + Cry34/35Ab1 maize in Nebraska.

BACKGROUND: Western corn rootworm (WCR; Diabrotica virgifera virgifera) field-evolved resistance to transgenic maize expressing the Cry3Bb1 protein derived from Bacillus thuringiensis (Bt) has been confirmed across the United States Corn Belt. Although use of pyramided hybrids expressing Cry3Bb1 + Cry34/35Ab1 has increased in recent years to mitigate existing WCR Bt resistance, susceptibility of Nebraska WCR populations to this rootworm-Bt pyramid has not been assessed. Plant-based bioassays were used to characterize the susceptibility of WCR populations to Cry3Bb1 and Cry3Bb1 + Cry34/35Ab1 maize. Populations were collected from areas of northeastern Nebraska with a history of planting Bt maize that expressed Cry3Bb1 and Cry34/35Ab1. RESULTS: Significant differences in mean corrected survival among populations within Bt hybrids indicated a mosaic of WCR susceptibility to Cry3Bb1 + Cry34/35Ab1 and Cry3Bb1 maize occurred in the landscape. All field populations exhibited some level of resistance to one or both Bt hybrids when compared to susceptible laboratory control populations in bioassays. Most WCR populations exhibited incomplete resistance to Cry3Bb1 + Cry34/35Ab1 maize (92%) and complete resistance to Cry3Bb1 maize (79%). CONCLUSION: The present study confirms the first cases of field-evolved resistance to Cry3Bb1 + Cry34/35Ab1 maize in Nebraska and documents a landscape-wide WCR Cry3Bb1 resistance pattern in areas characterized by long-term continuous maize production and associated planting of Cry3Bb1 hybrids. Use of a multi-tactic integrated pest management approach is needed in areas of continuous maize production to slow or mitigate resistance evolution to Bt maize. © 2021 Society of Chemical Industry.

Up-regulation of apoptotic- and cell survival-related gene pathways following exposures of western corn rootworm to B. thuringiensis crystalline pesticidal proteins in transgenic maize roots.

BACKGROUND: Resistance of pest insect species to insecticides, including B. thuringiensis (Bt) pesticidal proteins expressed by transgenic plants, is a threat to global food security. Despite the western corn rootworm, Diabrotica virgifera virgifera, being a major pest of maize and having populations showing increasing levels of resistance to hybrids expressing Bt pesticidal proteins, the cell mechanisms leading to mortality are not fully understood. RESULTS: Twenty unique RNA-seq libraries from the Bt susceptible D. v. virgifera inbred line Ped12, representing all growth stages and a range of different adult and larval exposures, were assembled into a reference transcriptome. Ten-day exposures of Ped12 larvae to transgenic Bt Cry3Bb1 and Gpp34/Tpp35Ab1 maize roots showed significant differential expression of 1055 and 1374 transcripts, respectively, compared to cohorts on non-Bt maize. Among these, 696 were differentially expressed in both Cry3Bb1 and Gpp34/Tpp35Ab1 maize exposures. Differentially-expressed transcripts encoded protein domains putatively involved in detoxification, metabolism, binding, and transport, were, in part, shared among transcripts that changed significantly following exposures to the entomopathogens Heterorhabditis bacteriophora and Metarhizium anisopliae. Differentially expressed transcripts in common between Bt and entomopathogen treatments encode proteins in general stress response pathways, including putative Bt binding receptors from the ATP binding cassette transporter superfamily. Putative caspases, pro- and anti-apoptotic factors, as well as endoplasmic reticulum (ER) stress-response factors were identified among transcripts uniquely up-regulated following exposure to either Bt protein. CONCLUSIONS: Our study suggests that the up-regulation of genes involved in ER stress management and apoptotic progression may be important in determining cell fate following exposure of susceptible D. v. virgifera larvae to Bt maize roots. This study provides novel insights into insect response to Bt intoxication, and a possible framework for future investigations of resistance mechanisms.

Characterizing the Relationship Between Western Corn Rootworm (Coleoptera: Chrysomelidae) Larval Survival on Cry3Bb1-Expressing Corn and Larval Development Metrics.

The western corn rootworm, Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae), is a significant pest of field corn, Zea mays L. (Poales: Poaceae), across the United States Corn Belt. Widespread adoption and continuous use of corn hybrids expressing the Cry3Bb1 protein to manage the western corn rootworm has resulted in greater than expected injury to Cry3Bb1-expressing hybrids in multiple areas of Nebraska. Single-plant bioassays were conducted on larval western corn rootworm populations to determine the level of resistance present in various Nebraska counties. The results confirmed a mosaic of susceptibility to Cry3Bb1 across Nebraska. Larval development metrics, including head capsule width and fresh weight, were measured to quantify the relationship between the level of resistance to Cry3Bb1 and larval developmental rate. Regression and correlation analyses indicate a significant positive relationship between Cry3Bb1 corrected survival and both larval development metrics. Results indicate that as the level of resistance to Cry3Bb1 within field populations increases, mean head capsule width and larval fresh weight also increase. This increases our understanding of western corn rootworm population dynamics and age structure variability present in the transgenic landscape that is part of the complex interaction of factors that drives resistance evolution. This collective variability and complexity within the landscape reinforces the importance of making corn rootworm management decisions based on information collected at the local level.

The Use of Insecticides to Manage the Western Corn Rootworm, Diabrotica virgifera virgifera, LeConte: History, Field-Evolved Resistance, and Associated Mechanisms.

The western corn rootworm, Diabrotica virgifera virgifera LeConte (Dvv) is a significant insect pest of maize in the United States (U.S.). This paper reviews the history of insecticide use in Dvv management programs, Dvv adaptation to insecticides, i.e., field-evolved resistance and associated mechanisms of resistance, plus the current role of insecticides in the transgenic era. In the western U.S. Corn Belt where continuous maize is commonly grown in large irrigated monocultures, broadcast-applied soil or foliar insecticides have been extensively used over time to manage annual densities of Dvv and other secondary insect pests. This has contributed to the sequential occurrence of Dvv resistance evolution to cyclodiene, organophosphate, carbamate, and pyrethroid insecticides since the 1950s. Mechanisms of resistance are complex, but both oxidative and hydrolytic metabolism contribute to organophosphate, carbamate, and pyrethroid resistance facilitating cross-resistance between insecticide classes. History shows that Dvv insecticide resistance can evolve quickly and may persist in field populations even in the absence of selection. This suggests minimal fitness costs associated with Dvv resistance. In the transgenic era, insecticides function primarily as complementary tools with other Dvv management tactics to manage annual Dvv densities/crop injury and resistance over time.

Molecular characterization of western corn rootworm pyrethroid resistance.

BACKGROUND: Western corn rootworm (WCR) pyrethroid resistance has been confirmed in the western US Corn Belt. Toxicological and biochemical studies indicated that multiple mechanisms of resistance might be involved in the resistance trait, such as enhanced metabolism and/or kdr target-site mutation(s) in the voltage-gated sodium channels. To characterize the mechanisms of WCR pyrethroid resistance at the molecular level, pairwise comparisons were made between RNA-Seq data collected from pyrethroid-resistant and -susceptible WCR populations. Gene expression levels and sodium channel sequences were evaluated. RESULTS: Seven transcripts exhibited significantly different expression (q ≤ 0.05) when comparing field-collected pyrethroid-resistant (R-Field) and -susceptible (S-Field) WCR populations. Three of the differentially expressed transcripts were P450s overexpressed in R-Field (9.2-26.2-fold). A higher number (99) of differentially expressed transcripts was found when comparing laboratory-derived pyrethroid-resistant (R-Lab) and -susceptible (S-Lab) WCR populations. Eight of the significant transcripts were P450s overexpressed in R-Lab (2.7-39.8-fold). This study did not detect kdr mutations in pyrethroid-resistant WCR populations. Other differentially expressed transcripts that may play a role in WCR pyrethroid resistance are discussed. CONCLUSION: This study revealed that P450-mediated metabolism is likely to be a major mechanism of WCR pyrethroid resistance, which could affect the efficacy of other insecticides sharing similar metabolic pathways. Additionally, results suggested that although laboratory selection of a pyrethroid-resistant WCR population may help to characterize resistance mechanisms, a field-selected population provided rare and perhaps major variants corresponding to the resistance trait.

Enhanced metabolism and selection of pyrethroid-resistant western corn rootworms (Diabrotica virgifera virgifera LeConte).

Western corn rootworm (WCR) pyrethroid resistance has been previously reported in the United States (US) western Corn Belt, and cross-resistance and synergism studies suggested that both target site insensitivity and enhanced metabolism may be conferring WCR resistance to pyrethroids. The present study aimed to investigate the potential mechanisms of WCR pyrethroid resistance and to estimate the heritability of the resistance trait. Biochemical assays using model substrates and spectrophotometry revealed 2-4-fold higher activity of P450s and esterases in pyrethroid-resistant WCR populations, whereas the biological activity of glutathione S-transferase was similar between populations tested. No mutation in the voltage-gated sodium channel was detected in pyrethroid-resistant WCR individuals by sequencing PCR products containing the para-homologous L1014, T929, and M918 amino acid positions that are commonly associated with target site mutations in other pyrethroid-resistant insects. A pilot estimation of pyrethroid resistance heritability obtained during laboratory selection of a WCR population suggested a major genetic component of the resistance trait and predicted a 10-fold increase in WCR bifenthrin resistance within ~7 generations of insecticide lethal exposure. Results support earlier indirect evidence that enhanced metabolism may be contributing to WCR resistance to pyrethroids and illustrates the potential of WCR pyrethroid resistance evolution.

Field efficacy of soil insecticides on pyrethroid-resistant western corn rootworms (Diabrotica virgifera virgifera LeConte).

BACKGROUND: Field-evolved pyrethroid resistance has been confirmed in western corn rootworm (WCR) populations collected from the United States (US) western Corn Belt. Resistance levels of WCR adults estimated in lab bioassays were confirmed to significantly reduce the efficacy of foliar-applied bifenthrin. The objective of the present study was to investigate the impact of WCR pyrethroid resistance levels on the performance of common soil-applied insecticide formulations (23.4% tefluthrin, 17.15% bifenthrin, and 0.1% cyfluthrin + 2.0% tebupirimphos). Field trials were conducted in 2016 and 2017 in three Nebraska, US, counties (Saunders, Clay, and Keith) where distinct levels of WCR susceptibility to pyrethroids (susceptible, moderately resistant, and highly resistant) had been previously reported in adult and larval bioassays. RESULTS: All soil insecticide treatments effectively protected maize roots from a pyrethroid-susceptible WCR population at Saunders. In contrast, the efficacy of bifenthrin and tefluthrin soil insecticides was significantly reduced at Clay and Keith, where pyrethroid-resistant WCR populations were reported. At Keith, where an additional failure of the cyfluthrin + tebupirimphos soil insecticide was observed, WCR laboratory dose-response bioassays showed a consistent ∼5-fold resistance level to the active ingredients bifenthrin, tefluthrin, and cyfluthrin. CONCLUSION: The efficacy of common soil insecticides used in the US for WCR management was significantly reduced in populations exhibiting relatively low levels of WCR pyrethroid resistance. Using a multitactical approach to manage WCR within an integrated pest management framework may mitigate resistance evolution and prolong the usefulness of WCR insecticides within the system. © 2019 Society of Chemical Industry.

Optimizing Egg Recovery From Wild Northern Corn Rootworm Beetles (Coleoptera: Chrysomelidae).

The northern corn rootworm, Diabrotica barberi Smith & Lawrence (Coleoptera: Chrysomelidae), is one of the most important insect pests in the U.S. Corn Belt. Efforts to obtain eggs from wild northern corn rootworm populations using techniques developed for other rootworm species have been unsuccessful due to lack of oviposition. In 2016, we evaluated four oviposition media in choice tests within each of three female densities in 30.5 × 30.5 × 30.5 cm BugDorm cages. The number of eggs laid per female was significantly affected by female density and the interaction of female density × oviposition media, but oviposition was relatively poor in all oviposition media (1.2 eggs per female when averaging the three female densities and all oviposition media). Single females were also evaluated in nonchoice assays in 6 cm × 6 cm × 8 cm clear plastic boxes and averaged up to 108 eggs per female depending on the oviposition media. In 2017, the cumulative number of eggs laid per female in boxes with one female was not significantly different from the number of eggs laid per female in boxes with 3 females. In 2018, the cumulative number of eggs laid per female was not significantly different between female densities of 1, 3, 5, or 10 females per box. Total egg production per box therefore increased as female density increased. More than 27,000 wild northern corn rootworm eggs were collected from just 190 females when collected relatively early in the field season. We now have an efficient and robust system for obtaining eggs from wild northern corn rootworm females.

Western corn rootworm pyrethroid resistance confirmed by aerial application simulations of commercial insecticides.

The western corn rootworm (Diabrotica virgifera virgifera LeConte) (WCR) is a major insect pest of corn (Zea mays L.) in the United States (US) and is highly adaptable to multiple management tactics. A low level of WCR field-evolved resistance to pyrethroid insecticides has been confirmed in the US western Corn Belt by laboratory dose-response bioassays. Further investigation has identified detoxification enzymes as a potential part of the WCR resistance mechanism, which could affect the performance of insecticides that are structurally related to pyrethroids, such as organophosphates. Thus, the responses of pyrethroid-resistant and -susceptible WCR populations to the commonly used pyrethroid bifenthrin and organophosphate dimethoate were compared in active ingredient bioassays. Results revealed a relatively low level of WCR resistance to both active ingredients. Therefore, a simulated aerial application bioassay technique was developed to evaluate how the estimated resistance levels would affect performance of registered rates of formulated products. The simulated aerial application technique confirmed pyrethroid resistance to formulated rates of bifenthrin whereas formulated dimethoate provided optimal control. Results suggest that the relationship between levels of resistance observed in dose-response bioassays and actual efficacy of formulated product needs to be further explored to understand the practical implications of resistance.

Southern Corn Rootworm (Coleoptera: Chrysomelidae) Adult Emergence and Population Growth Assessment After Selection With Vacuolar ATPase-A double-stranded RNA Over Multiple Generations.

The southern corn rootworm, Diabrotica undecimpunctata howardi Barber (Coleoptera: Chrysomelidae), was exposed over multiple generations to vacuolar (v)ATPase-A double-stranded (ds)RNA, first as adults and later, as neonate larvae. During adult selection, high mortality and lower fecundity were observed in the RNAi-selected cages after beetles were exposed to sublethal dsRNA concentrations that varied between LC40 and LC75. During larval selection, a delay in adult emergence and effects on population growth parameters were observed after neonates were exposed to sublethal dsRNA concentrations that varied between LC50 and LC70. Some of the parameters measured for adult emergence such as time to reach maximum linear adult emergence, time elapsed before attaining linear emergence, termination point of the linear emergence, and total days of linear emergence increase, were significantly different between RNAi-selected and control colonies for at least one generation. Significant differences were also observed in population growth parameters such as growth rate, net reproductive rate, doubling time, and generation time. After seven generations of selection, there was no indication that resistance evolved. The sublethal effects caused by exposures of southern corn rootworm to dsRNAs can affect important life history traits and fitness especially through delays in adult emergence and reduction in population growth. Although changes in susceptibility did not occur, the observation of sublethal effects suggests important responses to potential selection pressure. Assuming resistance involves a recessive trait, random mating between susceptible and resistant individuals is an important factor that allows sustainable use of transgenic plants, and delays in adult emergence observed in our studies could potentially compromise this assumption.

Spatial variation in western corn rootworm (Coleoptera: Chrysomelidae) susceptibility to Cry3 toxins in Nebraska.

Repeated use of field corn (Zea mays L.) hybrids expressing the Cry3Bb1 and mCry3A traits in Nebraska has selected for field-evolved resistance in some western corn rootworm (WCR; Diabrotica virgifera virgifera LeConte) populations. Therefore, this study was conducted to characterize spatial variation in local WCR susceptibility to Cry3Bb1 and mCry3A traits in Keith and Buffalo counties, Nebraska, and determine the relationship between past management practices and current WCR susceptibility. Adult WCR populations were collected from sampling grids during 2015 and 2016 and single-plant larval bioassays conducted with F1 progeny documented significant variation in WCR susceptibility to Cry3Bb1 and mCry3A on different spatial scales in both sampling grids. At the local level, results revealed that neighboring cornfields may support WCR populations with very different susceptibility levels, indicating that gene flow of resistant alleles from high trait survival sites is not inundating large areas. A field history index, comprised of additive and weighted variables including past WCR management tactics and agronomic practices, was developed to quantify relative selection pressure in individual fields. The field history index-Cry3 trait survivorship relationship from year 1 data was highly predictive of year 2 Cry3 trait survivorship when year 2 field history indices were inserted into the year 1 base model. Sensitivity analyses indicated years of trait use and associated selection pressure at the local level were the key drivers of WCR susceptibility to Cry3 traits in this system. Retrospective case histories from this study will inform development of optimal resistance management programs and increase understanding of plant-insect interactions that may occur when transgenic corn is deployed in the landscape. Results from this study also support current recommendations to slow or mitigate the evolution of resistance by using a multi-tactic approach to manage WCR densities in individual fields within an integrated pest management framework.

Seasonality of Colaspis crinicornis (Coleoptera: Chrysomelidae) and its Injury Potential to Corn in Southeastern Nebraska.

Colaspis crinicornis Schaeffer (Coleoptera: Chrysomelidae) primarily occurs in the Great Plains, United States. Although C. crinicornis has historically been considered a non-pest and is rarely found in agricultural systems, population densities of this species have been increasing in corn, Zea mays L., and soybean, Glycine max (L.) Merrill, over the last decade in southeastern Nebraska. As part of a comprehensive project to understand the life history and pest potential of C. crinicornis, a field study was conducted to: understand adult seasonality of C. crinicornis using emergence cages and whole-plant-count sampling in cornfields and sweep-net sampling in soybean fields; confirm voltinism and the overwintering stage; and evaluate the potential of larvae to cause economic injury to corn roots. Data indicate that C. crinicornis is univoltine in southeastern Nebraska and overwinters as medium-large larvae at least 20 cm deep in the soil. Adults were present from June through August with peak emergence in July. The C. crinicornis lifecycle is similar to related Colaspis species. Root injury to corn was minor at population densities encountered in the field, and therefore, C. crinicornis is unlikely to cause economic loss. C. crinicornis may be an example of an insect species that has exploited open niches in crops that have been created by changes in agricultural and pest management practices. The lifecycle and polyphagous nature of the insect, annual crop rotation, the shift to minimum tillage, and replacement of insecticides with Bacillus thuringiensis Berliner (Bt) traits may have collectively facilitated establishment and increased survival in agroecosystems.