Presence-Absence Sampling Plans for Stink Bugs (Hemiptera: Pentatomidae) in the Midwest Region of the United States.

Stink bugs represent an increasing risk to soybean production in the Midwest region of the United States. The current sampling protocol for stink bugs in this region is tailored for population density estimation and thus is more relevant to research purposes. A practical decision-making framework with more efficient sampling effort for management of herbivorous stink bugs is needed. Therefore, a binomial sequential sampling plan was developed for herbivorous stink bugs in the Midwest region. A total of 146 soybean fields were sampled across 11 states using sweep nets in 2016, 2017, and 2018. The binomial sequential sampling plans were developed using combinations of five tally thresholds at two proportion infested action thresholds to identify those that provided the best sampling outcomes. Final assessment of the operating characteristic curves for each plan indicated that a tally threshold of 3 stink bugs per 25 sweeps, and proportion infested action thresholds of 0.75 and 0.95 corresponding to the action thresholds of 5 and 10 stink bugs per 25 sweeps, provided the optimal balance between highest probability of correct decisions (≥ 99%) and lowest probability of incorrect decisions (≤ 1%). In addition, the average sample size for both plans (18 and 12 sets of 25 sweeps, respectively) was lower than that for the other proposed plans. The binomial sequential sampling plan can reduce the number of sample units required to achieve a management decision, which is important because it can potentially reduce risk/cost of management for stink bugs in soybean in this region.

Parasitism of Adult Pentatomidae by Tachinidae in Soybean in the North Central Region of the United States.

Stink bugs (Hemiptera: Pentatomidae) are agricultural pests of increasing significance in the North Central Region of the United States, posing a threat to major crops such as soybean. Biological control can reduce the need for insecticides to manage these pests, but the parasitism of stink bugs by Tachinidae (Diptera) is poorly characterized in this region. The objective of this study was to evaluate the rate of parasitism of stink bugs by tachinids over 2 yr from nine states across the North Central Region. Parasitism was assessed by quantifying tachinid eggs on the integument of stink bug adults. Parasitism rates (i.e., percent of adult stink bugs with tachinid eggs) were compared across stink bug species, states, stink bug sex, and years. The mean percent parasitism of stink bugs by tachinids was about 6% across the region and did not differ among stink bug species. Mean percent parasitism was significantly higher in Missouri than in northern and western states. In addition, male stink bugs had significantly higher mean percent parasitism than females. Stink bug species commonly found in soybean in the region showed some parasitism and are therefore potentially vulnerable to oviposition by these parasitoids. This is the first study to characterize the level of parasitism of stink bugs by tachinids across the North Central Region.

Implementation of a Diagnostic-Concentration Bioassay for Detection of Susceptibility to Pyrethroids in Soybean Aphid (Hemiptera: Aphididae).

Soybean aphid, Aphis glycines Matsumura, remains the most economically damaging arthropod pest of soybean in the midwestern United States and southern Canada. Foliar applications of a limited number of insecticide modes of action have been the primary management tactic, and pyrethroid resistance was documented recently with full concentration-response leaf-dip and glass-vial bioassays. Full concentration-response bioassays can be cumbersome, and a more efficient assessment tool was needed. In this study, we implemented a diagnostic-concentration glass-vial bioassay using bifenthrin and λ-cyhalothrin. Bioassays were conducted with field-collected soybean aphid populations to assess the geographic extent and severity of resistance to pyrethroids. In 2017, 10 of 18 and 11 of 21 field populations tested with bifenthrin and λ-cyhalothrin, respectively, had mean proportion mortalities less than the susceptible laboratory population. In 2018, 17 of 23 and 13 of 23 field populations tested with bifenthrin and λ-cyhalothrin, respectively, had mean proportion mortalities less than the susceptible laboratory population. Populations collected after reported field failures of a pyrethroid insecticide generally had mean proportion mortalities less than the susceptible laboratory population. In both years, there was a strong correlation between chemistries, which suggests cross-resistance between these insecticides. The diagnostic-concentration glass-vial bioassays reported here will provide the foundation for an insecticide resistance monitoring program with the ability to determine practical levels and geographic extent of insecticide resistance.

Molecular Basis of Soybean Resistance to Soybean Aphids and Soybean Cyst Nematodes.

Soybean aphid (SBA; Aphis glycines Matsumura) and soybean cyst nematode (SCN; Heterodera glycines Ichninohe) are major pests of the soybean (Glycine max [L.] Merr.). Substantial progress has been made in identifying the genetic basis of limiting these pests in both model and non-model plant systems. Classical linkage mapping and genome-wide association studies (GWAS) have identified major and minor quantitative trait loci (QTLs) in soybean. Studies on interactions of SBA and SCN effectors with host proteins have identified molecular cues in various signaling pathways, including those involved in plant disease resistance and phytohormone regulations. In this paper, we review the molecular basis of soybean resistance to SBA and SCN, and we provide a synthesis of recent studies of soybean QTLs/genes that could mitigate the effects of virulent SBA and SCN populations. We also review relevant studies of aphid-nematode interactions, particularly in the soybean-SBA-SCN system.

Neonicotinoid seed treatments of soybean provide negligible benefits to US farmers.

Neonicotinoids are the most widely used insecticides worldwide and are typically deployed as seed treatments (hereafter NST) in many grain and oilseed crops, including soybeans. However, there is a surprising dearth of information regarding NST effectiveness in increasing soybean seed yield, and most published data suggest weak, or inconsistent yield benefit. The US is the key soybean-producing nation worldwide and this work includes soybean yield data from 194 randomized and replicated field studies conducted specifically to evaluate the effect of NSTs on soybean seed yield at sites within 14 states from 2006 through 2017. Here we show that across the principal soybean-growing region of the country, there are negligible and management-specific yield benefits attributed to NSTs. Across the entire region, the maximum observed yield benefits due to fungicide (FST = fungicide seed treatment) + neonicotinoid use (FST + NST) reached 0.13 Mg/ha. Across the entire region, combinations of management practices affected the effectiveness of FST + NST to increase yield but benefits were minimal ranging between 0.01 to 0.22 Mg/ha. Despite widespread use, this practice appears to have little benefit for most of soybean producers; across the entire region, a partial economic analysis further showed inconsistent evidence of a break-even cost of FST or FST + NST. These results demonstrate that the current widespread prophylactic use of NST in the key soybean-producing areas of the US should be re-evaluated by producers and regulators alike.

Transcriptome profiling of interaction effects of soybean cyst nematodes and soybean aphids on soybean.

Soybean aphid (Aphis glycines; SBA) and soybean cyst nematode (Heterodera glycines; SCN) are two major pests of soybean (Glycine max) in the United States of America. This study aims to characterize three-way interactions among soybean, SBA, and SCN using both demographic and genetic datasets. SCN-resistant and SCN-susceptible soybean cultivars with a combination of soybean aphids (biotype 1) and SCN (HG type 0) in a randomized complete block design (RCBD) with six blocks were used to evaluate the three-way interactions in a greenhouse setup. Treatments receiving SCN were infested at planting with 2000 nematode eggs, and the treatments with soybean aphids were infested at second trifoliate growth stage (V2) with 15 soybean aphids. The whole roots were sampled from plants at 5 and 30 days post SBA infestation for RNA sequencing using Illumina Hiseq. 3000. The data comprises of 47 libraries that are useful for further analyses of important genes, which are involved in interaction effects of SBA and SCN on soybean.

Transcriptome profiling of induced susceptibility effects on soybean-soybean aphid (Hemiptera: Aphididae) interaction.

OBJECTIVES: Soybean aphid (Aphis glycines Matsumura; SBA) is the most economically damaging insect of soybean (Glycine max) in the United States. One previous study demonstrated that avirulent (biotype 1) and virulent (biotype 2) biotypes could co-occur and interact on resistant (i.e., Rag1) and susceptible soybean resulting in induced susceptibility after 11 days of feeding. The main objective of this research was to employ RNA sequencing (RNA-seq) technique to compare the induced susceptibility effect of biotype 2 on susceptible and resistant soybean at day 1 and day 11 (i.e., both susceptible and resistant soybean were initially challenged by biotype 2 and the effect was monitored through biotype 1 populations). DATA DESCRIPTION: We investigated susceptible and Rag1 transcriptome response to SBA feeding in soybean plants colonized by biotype 1 in the presence or absence of an inducer population (i.e., biotype 2). Ten RNA datasets are reported with 266,535,654 sequence reads (55.2 GB) obtained from pooled samples derived from the leaves collected at day 1 and day 11 post SBA infestation. A comprehensive understanding of these transcriptome data will enhance our understanding of interactions among soybean and two different biotypes of soybean aphids at the molecular level.

Community Composition, Abundance, and Phenology of Stink Bugs (Hemiptera: Pentatomidae) in Soybean in the North Central Region of the United States.

Stink bugs (Hemiptera: Pentatomidae) are an increasing threat to soybean (Fabales: Fabaceae) production in the North Central Region of the United States, which accounts for 80% of the country's total soybean production. Characterization of the stink bug community is essential for development of management programs for these pests. However, the composition of the stink bug community in the region is not well defined. This study aimed to address this gap with a 2-yr, 9-state survey. Specifically, we characterized the relative abundance, richness, and diversity of taxa in this community, and assessed phenological differences in abundance of herbivorous and predatory stink bugs. Overall, the stink bug community was dominated by Euschistus spp. (Hemiptera: Pentatomidae) and Chinavia hilaris (Say) (Hemiptera: Pentatomidae). Euschistus variolarius (Palisot de Beauvois) (Hemiptera: Pentatomidae), C. hilaris and Halyomorpha halys (Stål) (Hemiptera: Pentatomidae) were more abundant in the northwestern, southeastern and eastern parts, respectively, of the North Central Region of the United States. Economically significant infestations of herbivorous species occurred in fields in southern parts of the region. Species richness differed across states, while diversity was the same across the region. Herbivorous and predatory species were more abundant during later soybean growth stages. Our results represent the first regional characterization of the stink bug community in soybean fields and will be fundamental for the development of state- and region-specific management programs for these pests in the North Central Region of the United States.

Spatial Patterns and Sequential Sampling Plans for Estimating Densities of Stink Bugs (Hemiptera: Pentatomidae) in Soybean in the North Central Region of the United States.

Stink bugs are an emerging threat to soybean (Fabales: Fabaceae) in the North Central Region of the United States. Consequently, region-specific scouting recommendations for stink bugs are needed. The aim of this study was to characterize the spatial pattern and to develop sampling plans to estimate stink bug population density in soybean fields. In 2016 and 2017, 125 fields distributed across nine states were sampled using sweep nets. Regression analyses were used to determine the effects of stink bug species [Chinavia hilaris (Say) (Hemiptera: Pentatomidae) and Euschistus spp. (Hemiptera: Pentatomidae)], life stages (nymphs and adults), and field locations (edge and interior) on spatial pattern as represented by variance-mean relationships. Results showed that stink bugs were aggregated. Sequential sampling plans were developed for each combination of species, life stage, and location and for all the data combined. Results for required sample size showed that an average of 40-42 sample units (sets of 25 sweeps) would be necessary to achieve a precision of 0.25 for stink bug densities commonly encountered across the region. However, based on the observed geographic gradient of stink bug densities, more practical sample sizes (5-10 sample units) may be sufficient in states in the southeastern part of the region, whereas impractical sample sizes (>100 sample units) may be required in the northwestern part of the region. Our findings provide research-based sampling recommendations for estimating densities of these emerging pests in soybean.

Resistance of Soybean Plant Introductions to Three Colonies of Soybean Aphid (Hemiptera: Aphididae) Biotype 4.

Soybean aphid, Aphis glycines Matsumura (Hemiptera: Aphididae), infestations of soybean, Glycine max (L.) Merr. (Fabales: Fabaceae), and the associated yield loss have led to a large dependence on insecticidal management in soybean throughout the Midwestern United States. However, several populations of pyrethroid-resistant soybean aphids have recently been found in Iowa, Minnesota, North Dakota and South Dakota, which highlights the importance of alternative management approaches. One such alternative method is host-plant resistance, which uses naturally occurring plant defenses in crop cultivars to reduce the potential for yield loss from a pest population. Current soybean aphid-resistant cultivars do not protect against all soybean aphids due to the presence of virulent biotypes. In particular, soybean aphid biotype 4 is virulent to Rag1 and Rag2 resistance genes both individually and in combination. However, we hypothesized that resistance to biotype 4 may exist in previously identified, but uncharacterized resistant soybean plant introductions (PIs). To test this, we evaluated 51 previously identified but uncharacterized soybean aphid-resistant PIs for their resistance to colonies of soybean aphid biotype 4 collected in separate site-years (Lomira, WI 2013; Volga, SD 2015, 2016). Free-choice tests identified 14 PIs with putative resistance to 'Lomira13', two to 'Volga15', and eight to 'Volga16' soybean aphid colonies. Follow-up, no-choice tests corroborated two to three resistant PIs per colony, and PI 437696, which was resistant to each of the three colonies and could aid in breeding efforts and an integrated approach to soybean aphid management.

Native Solitary Bees Provide Economically Significant Pollination Services to Confection Sunflowers (Helianthus annuus L.) (Asterales: Asteraceae) Grown Across the Northern Great Plains.

The benefits of insect pollination to crop yields depend on genetic and environmental factors including plant self-fertility, pollinator visitation rates, and pollinator efficacy. While many crops benefit from insect pollination, such variation in pollinator benefits across both plant cultivars and growing regions is not well documented. In this study, across three states in the northern Great Plains, United States, from 2016 to 2017, we evaluated the pollinator-mediated yield increases for 10 varieties of confection sunflowers, Helianthus annuus L. (Asterales: Asteraceae), a plant that is naturally pollinator-dependent but was bred for self-fertility. We additionally measured pollinator visitation rates and compared per-visit seed set across pollinator taxa in order to determine the most efficacious sunflower pollinators. Across all locations and hybrids, insect pollination increased sunflower yields by 45%, which is a regional economic value of over $40 million and a national value of over $56 million. There was, however, some variation in the extent of pollinator benefits across locations and plant genotypes, and such variation was significantly related to pollinator visitation rates, further highlighting the value of pollinators for confection sunflowers. Female Andrena helianthi Robertson (Hymenoptera: Andrenidae) and Melissodes spp. (Hymenoptera: Apidae) were the most common and effective pollinators, while other bees including managed honey bees (Hymenoptera: Halictidae), Apis mellifera L. (Hymenoptera: Apidae), small-bodied sweat bees (Hymenoptera: Halictidae), bumble bees Bombus spp. (Hymenoptera: Apidae), and male bees were either infrequent or less effective on a per-visit basis. Our results illustrate that wild bees, in particular the sunflower specialists A. helianthi and Melissodes spp., provide significant economic benefits to confection sunflower production.

Rapid evolution to host plant resistance by an invasive herbivore: soybean aphid (Aphis glycines) virulence in North America to aphid resistant cultivars.

Preventing rapid evolution of herbivores to plant traits that confer resistance is an area of active research for applied entomologists. The subfield of insect resistance management (IRM) uses elements of population genetics and ecology to prevent increases in the frequency of virulent (i.e. resistant) sub-populations of an insect pest. Efforts to delay such an increase include using highly lethal toxins (i.e., a high dose), combining multiple resistance traits in one cultivar (i.e., pyramids), and using susceptible plants (i.e. a refuge) within or near plantings of the resistant crop. Even if fully implemented, theoretical models suggest that IRM plans for asexually-reproducing insects (e.g. aphids) cannot limit the frequency of resistance to provide sustainable use of a pest-resistant cultivar. We discuss how feeding by conspecifics aphids induces susceptibility such that a "within plant" refuge is created, allowing both virulent and avirulent (i.e. susceptible) populations to persist. We use the soybean aphid (Aphis glycines Matsumura), and the rapid occurrence of virulence in the US to resistant cultivars of soybean (Glycine max). We describe how feeding by A. glycines on soybeans alters the quality of the plant as a host. These systemic changes to the plants' physiology allow avirulent A. glycines to thrive on resistant cultivars. We explore how the induction of susceptibility by a herbivore can slow an increase in the frequency of virulent populations to resistant host plants. We suggest that a within plant refuge, combined with standard IRM practices, can allow for sustainable use of plant resistance to asexually-reproducing insect pests.

Evolutionary Divergence of TNL Disease-Resistant Proteins in Soybean (Glycine max) and Common Bean (Phaseolus vulgaris).

Disease-resistant genes (R genes) encode proteins that are involved in protecting plants from their pathogens and pests. Availability of complete genome sequences from soybean and common bean allowed us to perform a genome-wide identification and analysis of the Toll interleukin-1 receptor-like nucleotide-binding site leucine-rich repeat (TNL) proteins. Hidden Markov model (HMM) profiling of all protein sequences resulted in the identification of 117 and 77 regular TNL genes in soybean and common bean, respectively. We also identified TNL gene homologs with unique domains, and signal peptides as well as nuclear localization signals. The TNL genes in soybean formed 28 clusters located on 10 of the 20 chromosomes, with the majority found on chromosome 3, 6 and 16. Similarly, the TNL genes in common bean formed 14 clusters located on five of the 11 chromosomes, with the majority found on chromosome 10. Phylogenetic analyses of the TNL genes from Arabidopsis, soybean and common bean revealed less divergence within legumes relative to the divergence between legumes and Arabidopsis. Syntenic blocks were found between chromosomes Pv10 and Gm03, Pv07 and Gm10, as well as Pv01 and Gm14. The gene expression data revealed basal level expression and tissue specificity, while analysis of available microRNA data showed 37 predicted microRNA families involved in targeting the identified TNL genes in soybean and common bean.

Reduced Insecticide Susceptibility in Aedes vexans (Diptera: Culicidae) Where Agricultural Pest Management Overlaps With Mosquito Abatement.

Mosquito abatement programs in Midwestern communities frequently exist within landscapes dominated by agriculture. Although separately managed, both agricultural pests and mosquitoes are targeted by similar classes of insecticides. As a result, there is the potential for unintended insecticide exposure to mosquito populations from agricultural pest management. To determine the impact that agricultural management practices have on mosquito insecticide susceptibility we compared the mortality of Aedes vexans (Meigen; Diptera: Culicidae) between populations sampled from locations with and without mosquito abatement in South Dakota, a region dominated by agricultural production. Collection locations were either within towns with mosquito abatement programs (n = 2; Brookings and Sioux Falls, SD) or located > 16 km from towns with mosquito abatement programs (n = 2; areas near Harrold and Willow Lake, SD). WHO bioassays were used to test susceptibly of adults to differing insecticide classes relative to their respective controls; 1) an organochlorine (dieldrin 4%), 2) an organophosphate (malathion 5%), and 3) a pyrethroid (lambda-cyhalothrin 0.05%). Corrected mortality did not significantly differ between locations with or without abatement; however, when locations were analized by proportion of developed land within the surrounding landscape pyrethroid mortality was significantly lower where crop production dominated the surrounding landscape and mosquito abatement was present. These data suggest that agricultural pest management may incidentally contribute to reduced mosquito susceptibility where overlap between agricultural pest management and mosquito abatement exists. Decoupling insecticide classes used by both agricultural and public health pest management programs may be necessary to ensure continued efficacy of pest management tools.

Reduced Fitness of Virulent Aphis glycines (Hemiptera: Aphididae) Biotypes May Influence the Longevity of Resistance Genes in Soybean.

Sustainable use of insect resistance in crops require insect resistance management plans that may include a refuge to limit the spread of virulence to this resistance. However, without a loss of fitness associated with virulence, a refuge may not prevent virulence from becoming fixed within a population of parthenogenetically reproducing insects like aphids. Aphid-resistance in soybeans (i.e., Rag genes) prevent outbreaks of soybean aphid (Aphis glycines), yet four biotypes defined by their capacity to survive on aphid-resistant soybeans (e.g., biotype-2 survives on Rag1 soybean) are found in North America. Although fitness costs are reported for biotype-3 on aphid susceptible and Rag1 soybean, it is not clear if virulence to aphid resistance in general is associated with a decrease in fitness on aphid susceptible soybeans. In laboratory assays, we measured fitness costs for biotype 2, 3 and 4 on an aphid-susceptible soybean cultivar. In addition, we also observed negative cross-resistance for biotype-2 on Rag3, and biotype-3 on Rag1 soybean. We utilized a simple deterministic, single-locus, four compartment genetic model to account for the impact of these findings on the frequency of virulence alleles. When a refuge of aphid susceptible was included within this model, fitness costs and negative cross-resistance delayed the increase of virulence alleles when virulence was inherited recessively or additively. If virulence were inherited additively, fitness costs decreased the frequency of virulence. Combined, these results suggest that a refuge may prevent virulent A. glycines biotypes from overcoming Rag genes if this aphid-resistance were used commercially in North America.