Best Management Practices for Corn Production in South Dakota: Corn Insect Pests

Historically, the major corn insect pests have been corn rootworms (northern and western), European corn borer, and black cutworm. Bt-corn hybrids are effective against most of these pests. However, Bt-corn hybrids are not effective against corn leaf aphid, corn root aphid, sap beetles, corn rootworm adults, grasshoppers, white grubs, wireworms, seed corn beetle, and seed corn maggots. These insect pests can reduce corn yields. This chapter discusses the management and biology of important corn insect pests commonly observed in South Dakota

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

Utility of EST-Derived SSRs as Population Genetics Markers in a Beetle

Microsatellite, or simple sequence repeat (SSR), loci can be identified by mining expressed sequence tag (EST) databases, and where these are available, marker development time and expense can be decreased considerably over conventional strategies of probing the entire genome. However, it is unclear whether they provide information on population structure similar to that generated by anonymous genomic SSRs. We performed comparative population genetic analyses between EST-derived SSRs (EST-SSRs) and anonymous SSRs developed from genomic DNA for the same set of populations of the insect Diabrotica virgifera, a beetle in the family Chrysomelidae. Compared with noncoding, nontranscribed regions, EST-SSRs were generally less polymorphic but had reduced occurrence of null alleles and greater cross-species amplification. Neutrality tests suggested the loci were not under positive selection. Across all populations and all loci, the genomic and EST-SSRs performed similarly in estimating genetic diversity, FIS, FST, population assignment and exclusion tests, and detection of distinct populations. These findings, therefore, indicate that the EST-SSRs examined can be used with confidence in future genetic studies of Diabrotica populations and suggest that EST libraries can be added as a valuable source of markers for population genetics studies in insects and other animals

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

Toxicity of insecticide-bait mixtures to insecticide resistant and susceptible western corn rootworms (Coleoptera: Chrysomelidae)

Organophosphate resistant and susceptible populations of the western corn rootworm, Diabrotica virgifera virgifera LeConte. were subjected to adult feeding bioassays with different combinations of insecticide and a cucurbitacin bait. Five technical grade insecticides (methyl-parathion, carbaryl, fipronil, bifenthrin and indoxacarb) were used in combination with Invite ECTM as the feeding stimulant. Differences in susceptibility to the insecticide/bait combinations were observed among the resistant and susceptible populations for methyl-parathion and carbaryl. Susceptibility to fipronil, bifenthrin and indoxacarb was similar among the resistant and susceptible populations. Assays in which response to the insecticide/bait combination was compared with the bait alone indicated that methyl-parathion and bifenthrin were deterrent when compared to other treatments. These results suggest that the efficacy of Invite as a feeding stimulant in combination with certain insecticides may be compromised by previously identified resistance and by insecticides that antagonize the feeding stimulation of the cucurbitacin bait

Distribution of Genes and Repetitive Elements in the Diabrotica virgifera virgifera Genome Estimated Using BAC Sequencing

Feeding damage caused by the western corn rootworm, Diabrotica virgifera virgifera, is destructive to corn plants in North America and Europe where control remains challenging due to evolution of resistance to chemical and transgenic toxins. A BAC library, DvvBAC1, containing 109,486 clones with 1 0 4 ± 3 4 . 5 kb inserts was created, which has an ~4.56X genome coverage based upon a 2.58 Gb (2.80 pg) flow cytometry-estimated haploid genome size. Paired end sequencing of 1037 BAC inserts produced 1.17 Mb of data (~0.05% genome coverage) and indicated ~9.4 and 16.0% of reads encode, respectively, endogenous genes and transposable elements (TEs). Sequencing genes within BAC full inserts demonstrated that TE densities are high within intergenic and intron regions and contribute to the increased gene size. Comparison of homologous genome regions cloned within different BAC clones indicated that TE movement may cause haplotype variation within the inbred strain. The data presented here indicate that the D. virgifera virgifera genome is large in size and contains a high proportion of repetitive sequence. These BAC sequencing methods that are applicable for characterization of genomes prior to sequencing may likely be valuable resources for genome annotation as well as scaffolding

Comparison of Six Artificial Diets for Western Corn Rootworm Bioassays and Rearing

The western corn rootworm, Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae), is considered the most important maize (Zea mays L.) pest in the U.S. Corn Belt. Bioassays testing susceptibility to Bacillus thuringiensis Berliner (Bt) and other toxins of corn rootworm larvae often rely on artificial diet formulations. Successful bioassays on artificial diet for corn rootworm have sometimes been challenging because of microbial contamination. Toward the long-term goal of developing a universal artificial diet for western corn rootworm larvae, we compared larval survival, dry weight, and percentage of molt in 10-d bioassays from six current diets of which we were aware. In addition, as part of longer term rearing efforts, we recorded molting over an extended period of development (60 d). Six different artificial diets, including four proprietary industry diets (A, B, C, and D), the first published artificial diet for western corn rootworm (Pleau), and a new diet (WCRMO-1) were evaluated. Western corn rootworm larval survival was above 90% and contamination was 0% on all diets for 10 d. Diet D resulted in the greatest dry weight and percentage molting when compared with the other diets. Although fourth-instar western corn rootworm larvae have not been documented previously (only three instars have been previously documented), as many as 10% of the larvae from Diet B molted into a fourth instar prior to pupating. Overall, significant differences were found among artificial diets currently used to screen western corn rootworm. In order for data from differing toxins to be compared, a single, reliable and high-quality western corn rootworm artificial diet should eventually be chosen by industry, academia, and the public as a standard for bioassays

Evidence of Resistance to Cry34/35Ab1 Corn by Western Corn Rootworm (Coleoptera: Chrysomelidae): Root Injury in the Field and Larval Survival in Plant-Based Bioassays

Western corn rootworm, Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae), is a serious pest of corn in the United States, and recent management of western corn rootworm has included planting of Bt corn. Beginning in 2009, western corn rootworm populations with resistance to Cry3Bb1 corn and mCry3A corn were found in Iowa and elsewhere. To date, western corn rootworm populations have remained susceptible to corn producing Bt toxin Cry34/35Ab1. In this study, we used single-plant bioassays to test field populations of western corn rootworm for resistance to Cry34/35Ab1 corn, Cry3Bb1 corn, and mCry3A corn. Bioassays included nine rootworm populations collected from fields where severe injury to Bt corn had been observed and six control populations that had never been exposed to Bt corn. We found incomplete resistance to Cry34/35Ab1 corn among field populations collected from fields where severe injury to corn producing Cry34/35Ab1, either singly or as a pyramid, had been observed. Additionally, resistance to Cry3Bb1 corn and mCry3A corn was found among the majority of populations tested. These first cases of resistance to Cry34/35Ab1 corn, and the presence of resistance to multiple Bt toxins by western corn rootworm, highlight the potential vulnerability of Bt corn to the evolution of resistance by western corn rootworm. The use of more diversified management practices, in addition to insect resistance management, likely will be essential to sustain the viability of Bt corn for management of western corn rootworm