Landscape-level variation in Bt crops predict Helicoverpa zea (Lepidoptera: Noctuidae) resistance in cotton agroecosystems.

BACKGROUND: Helicoverpa zea (Boddie) damage to Bt cotton and maize has increased as a result of widespread Bt resistance across the USA Cotton Belt. Our objective was to link Bt crop production patterns to cotton damage through a series of spatial and temporal surveys of commercial fields to understand how Bt crop production relates to greater than expected H. zea damage to Bt cotton. To do this, we assembled longitudinal cotton damage data that spanned the Bt adoption period, collected cotton damage data since Bt resistance has been detected, and estimated local population susceptibility using replicated on-farm studies that included all Bt pyramids marketed in cotton. RESULTS: Significant year effects of H. zea damage frequency in commercial cotton were observed throughout the Bt adoption period, with a recent damage increase after 2012. Landscape-level Bt crop production intensity over time was positively associated with the risk of H. zea damage in two- and three-toxin pyramided Bt cotton. Helicoverpa zea damage also varied across Bt toxin types in spatially replicated on-farm studies. CONCLUSIONS: Landscape-level predictors of H. zea damage in Bt cotton can be used to identify heightened Bt resistance risk areas and serves as a model to understand factors that drive pest resistance evolution to Bt toxins in the southeastern United States. These results provide a framework for more effective insect resistance management strategies to be used in combination with conventional pest management practices that improve Bt trait durability while minimizing the environmental footprint of row crop agriculture. © 2021 Society of Chemical Industry. This article has been contributed to by US Government employees and their work is in the public domain in the USA.

Screening Germplasm and Quantification of Components Contributing to Thrips Resistance in Cotton.

Three hundred and ninety-one Gossypium hirsutum and 34 Gossypium barbadense accessions were screened for thrips resistance under field conditions at the Upper Coastal Plain Research Station in Rocky Mount, North Carolina in years 2014 and 2015. Visual damage ratings, thrips counts, and seedling dry weights were recorded at 2.5, 3.5, and 4.5 wk after planting, respectively. Population density and thrips arrival times varied between years. Data from the three separate damage scoring dates provided a better estimate of resistance or susceptibility to thrips than ratings from the individual dates over the season. Tobacco thrips [Frankliniella fusca (Hinds) (Thysanoptera: Thripidae)], followed by western flower thrips [Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae)], were the dominant thrips species observed in the study. Five resistant G. barbadense accessions and five moderately resistant upland cotton accessions were identified from field evaluations. Greenhouse experiments were conducted in Fall 2015 and Spring 2016 to determine if plant height, growth rate, leaf pubescence, and leaf area were significantly different in resistant and susceptible groups of G. hirsutum and G. barbadense accessions identified from the field screenings. Leaf pubescence and relative growth rate were significantly higher in resistant accessions compared with susceptible accessions in absence of thrips. There was no difference for plant height and leaf area between resistant and susceptible groups. Results suggest thrips-resistant plants have a possible competitive advantage through faster growth and higher trichome density, which limits thrips movement.

Long-Term Empirical and Observational Evidence of Practical Helicoverpa zea Resistance to Cotton With Pyramided Bt Toxins.

Evidence of practical resistance of Helicoverpa zea (Boddie) (Lepidoptera: Noctuidae) to Bt cotton in the United States is debatable, supported with occasional reports of boll damage in the field. Our objective was to provide both empirical and long-term observational evidence of practical resistance by linking both in-season and end-of-season measurements of H. zea damage to pyramided Bt cotton bolls and to provide Cry1Ac diet-based bioassay data in support of these damage estimates. In-season boll damage from H. zea was highly correlated to end-of-season damaged bolls. Across North Carolina, Bt cotton fields with end-of-season bolls damaged by H. zea increased during 2016 compared to previous years. Elevated damage was coupled with an increase in field sprays targeting H. zea during 2016, but not related to an increase in H. zea abundance. Bioassay data indicated that there was a range of Cry1Ac susceptibility across the southeastern United States. Given the range of susceptibility to Cry1Ac across the southeastern United States, it is probable that resistant populations are common. Since H. zea is resistant to cotton expressing pyramided Cry toxins, the adoption of new cotton varieties expressing Vip3Aa will be rapid. Efforts should be made to delay resistance of H. zea to the Vip3Aa toxin to avoid foliar insecticide use.

Cereal Leaf Beetle (Coleoptera: Chrysomelidae) Regional Dispersion and Relationship With Wheat Stand Denseness.

Cereal leaf beetle, Oulema melanopus L., is a pest of small grains and the literature conflicts on whether it is more abundant in sparse or dense stands of wheat. Our objectives were to determine the impact of stand denseness on cereal leaf beetle abundance and to investigate the regional dispersion of cereal leaf beetles across North Carolina and Virginia. One-hundred twenty fields were sampled across North Carolina and Virginia during 2011 for stand denseness, and cereal leaf beetle eggs, larvae, and adults. Two small-plot wheat experiments were planted in North Carolina using a low and a high seeding rate. Main plots were split, with one receiving a single nitrogen application and one receiving two. Egg density, but not larva or adult density, was positively correlated with stand denseness in the regional survey. Furthermore, regional spatial patterns of aggregation were noted for both stand denseness and egg number. In the small-plot experiments, seeding rate influenced stand denseness, but not nitrogen application. In one experiment, egg densities per unit area were higher in denser wheat, while in the other experiment, egg densities per tiller were lower in denser wheat. Larvae were not influenced by any factor. Overall, there were more cereal leaf beetle eggs in denser wheat stands. Previous observations that sparse stands of wheat are more prone to cereal leaf beetle infestation can be attributed to the fact that sparser stands have fewer tillers, which increases the cereal leaf beetle to tiller ratio compared with denser stands.

Does Florivory by Helicoverpa zea (Lepidoptera: Noctuidae) Cause Yield Loss in Soybeans?

Helicoverpa zea (Boddie), corn earworm, is a damaging insect pest of many crops, including soybeans. An economic threshold for soybeans during the pod-filling stages exists to prevent economic damage to seeds. However, the impact of florivory (flower feeding) by H. zea larvae on seed yield is poorly understood and there is no economic threshold for flowering-stage soybeans. Four small plot experiments were conducted in North Carolina during 2011 and 2012 to assess the impact of H. zea feeding during the flowering stages of determinate soybeans on various yield components. Helicoverpa zea densities were manipulated with insecticides and various planting dates of soybeans and monitored weekly. Helicoverpa zea naturally infested the plots after flowering began and were allowed to feed until R3; they were eliminated from all plots from R3 to maturity. In some sites, H. zea densities exceeded the podding economic threshold during the flowering stages, but yield did not differ among treatments. During 2012, florivory from H. zea was measured directly by counting injured flowers. There was a negative yield relationship between both injured flower number and cumulative flower number. Moreover, H. zea densities were related to both a decrease in cumulative flowers and an increase in injured flowers, even though a direct linkage between H. zea density and yield loss was not observed. Without knowing the preferred tissue types and performance of early-instar larvae on soybeans, it is possible that H. zea density may not be the best measurement for developing an economic threshold in flowering soybeans.

Impacts of Tillage, Maturity Group, and Insecticide Use on Megacopta cribraria (Hemiptera: Plataspidae) Populations in Double-Cropped Soybean.

Megacopta cribraria (F.), also known as the kudzu bug, is a soybean pest in the United States, and it can cause up to a 60% yield reduction if not controlled. Insecticides are commonly used to manage this pest in commercial soybean fields. However, other soybean production practices may also affect kudzu bug populations. This study investigated the effect of soil tillage, maturity group selection, and insecticide use on kudzu bug densities in soybean. During 2012 and 2013, at two locations each year in North Carolina, four varieties of soybean maturity groups were planted in June into conventionally tilled plots and into plots with cereal crop residue under reduced tillage conditions (mimicking double-crop production). Plots were further split as insecticide-protected and untreated. Four times more kudzu bugs were found in conventionally tilled than reduced till plots throughout the growing season. Selection of the maturity group influenced the attractiveness of the kudzu bug to oviposit on soybean. A 56% reduction of kudzu bug densities was achieved through insecticide treatment, with an ∼6% increase in yield. Information on how production practices, including soil tillage, affect kudzu bug populations in soybean may help growers select practices to minimize kudzu bug injury and protect yield.

Biology, Pest Status, Microbiome and Control of Kudzu Bug (Hemiptera: Heteroptera: Plataspidae): A New Invasive Pest in the U.S.

Soybean is an important food crop, and insect integrated pest management (IPM) is critical to the sustainability of this production system. In recent years, the introduction into the United States of the kudzu bug currently identified as Megacopta cribraria (F.), poses a threat to soybean production. The kudzu bug was first discovered in the state of Georgia, U.S. in 2009 and since then has spread to most of the southeastern states. Because it was not found in the North American subcontinent before this time, much of our knowledge of this insect comes from research done in its native habitat. However, since the U.S. introduction, studies have been undertaken to improve our understanding of the kudzu bug basic biology, microbiome, migration patterns, host selection and management in its expanding new range. Researchers are not only looking at developing IPM strategies for the kudzu bug in soybean, but also at its unique relationship with symbiotic bacteria. Adult females deposit bacterial packets with their eggs, and the neonates feed on these packets to acquire the bacteria, Candidatus Ishikawaella capsulata. The kudzu bug should be an informative model to study the co-evolution of insect function and behavior with that of a single bacteria species. We review kudzu bug trapping and survey methods, the development of bioassays for insecticide susceptibility, insecticide efficacy, host preferences, impact of the pest on urban environments, population expansion, and the occurrence of natural enemies. The identity of the kudzu bug in the U.S. is not clear. We propose that the kudzu bug currently accepted as M. cribraria in the U.S. is actually Megacopta punctatissima, with more work needed to confirm this hypothesis.

Megacopta cribraria (Hemiptera: Plataspidae) Population Dynamics in Soybeans as Influenced by Planting Date, Maturity Group, and Insecticide Use.

Since its unintentional introduction during 2009, Megacopta cribraria (F.) has spread rapidly throughout the southeastern United States, mainly feeding and reproducing on kudzu, Pueraria montana Loureiro (Merr.) variety lobata (Willdenow), and soybeans, Glycine max (L.) Merr. Megacopta cribraria has become a serious economic pest in soybeans, forcing growers to rely solely on insecticide applications to control this insect. The main objective of this study was to investigate if variation in planting date and maturity group of soybeans had an impact on management of M. cribraria populations. Three experimental fields were located in North Carolina (2) and South Carolina (1), and the tests replicated during 2012 and 2013. Treatments consisted of three planting dates, four maturity groups, and insecticide treated versus untreated, at each location. More M. cribraria were found in untreated early planted soybeans than late planted soybeans. Generally, maturity group did not influence population densities of M. cribraria. Yield was significantly influenced by the interaction between planting date and maturity group. There was a negative linear relationship between M. cribraria populations and soybean yield. Although early planted soybeans may avoid drought conditions and potentially large populations of defoliators, these fields may be at greater risk for infestation by M. cribraria.

Systemic Imidacloprid Affects Intraguild Parasitoids Differently.

Toxoneuron nigriceps (Viereck) (Hymenoptera, Braconidae) and Campoletis sonorensis (Cameron) (Hymenoptera, Ichneumonidae) are solitary endoparasitoids of the tobacco budworm, Heliothis virescens (Fabricius) (Lepidoptera, Noctuidae). They provide biological control of H. virescens populations in Southeastern US agricultural production systems. Field and greenhouse experiments conducted from 2011-2014 compared parasitism rates of parasitoids that developed inside H. virescens larvae fed on tobacco plants treated with and without imidacloprid. The parasitoids in our study did not have a similar response. Toxoneuron nigriceps had reduced parasitism rates, but parasitism rates of C. sonorensis were unaffected. Preliminary data indicate that adult female lifespans of T. nigriceps are also reduced. ELISA was used to measure concentrations of neonicotinoids, imidacloprid and imidacloprid metabolites in H. virescens larvae that fed on imidacloprid-treated plants and in the parasitoids that fed on these larvae. Concentrations were detectable in the whole bodies of parasitized H. virescens larvae, T. nigriceps larvae and T. nigriceps adults, but not in C. sonorensis larvae and adults. These findings suggest that there are effects of imidacloprid on multiple trophic levels, and that insecticide use may differentially affect natural enemies with similar feeding niches.

Relationship Between Piercing-Sucking Insect Control and Internal Lint and Seed Rot in Southeastern Cotton (Gossypium hirsutum L.).

In 1999, crop consultants scouting for stink bugs (Hemiptera spp.) in South Carolina discovered a formerly unobserved seed rot of cotton that caused yield losses ranging from 10 to 15% in certain fields. The disease has subsequently been reported in fields throughout the southeastern Cotton Belt. Externally, diseased bolls appeared undamaged; internally, green fruit contain pink to dark brown, damp, deformed lint, and necrotic seeds. In greenhouse experiments, we demonstrated transmission of the opportunistic bacterium Pantoea agglomerans by the southern green stink bug, Nezara viridula (L.). Here, green bolls were sampled from stink bug management plots (insecticide protected or nontreated) from four South Atlantic coast states (North Carolina, South Carolina, Georgia, and Florida) to determine disease incidence in the field and its association with piercing-sucking insects feeding. A logistic regression analysis of the boll damage data revealed that disease was 24 times more likely to occur (P = 0.004) in bolls collected from plots in Florida, where evidence of pest pressure was highest, than in bolls harvested in NC with the lowest detected insect pressure. Fruit from plots treated with insecticide, a treatment which reduced transmission agent numbers, were 4 times less likely to be diseased than bolls from unprotected sites (P = 0.002). Overall, punctured bolls were 125 times more likely to also have disease symptoms than nonpunctured bolls, irrespective of whether or not plots were protected with insecticides (P = 0.0001). Much of the damage to cotton bolls that is commonly attributed to stink bug feeding is likely the resulting effect of vectored pathogens.

Efficacy and value of prophylactic vs. integrated pest management approaches for management of cereal leaf beetle (Coleoptera: Chrysomelidae) in wheat and ramifications for adoption by growers.

Cereal leaf beetle, Oulema melanopus L., can be effectively managed in southeastern U.S. wheat, Triticum aestivum L., with scouting and a single insecticide treatment, applied at the recommended economic threshold. However, many growers eschew this approach for a prophylactic treatment, often tank mixed with a nitrogen application before wheat growth stage 30. The efficacy of a prophylactic and an integrated pest management (IPM) approach was compared for 2 yr using small plot studies in North Carolina and regional surveys across North Carolina and Virginia. Economic analyses were performed, comparing the total cost of management of each approach using the regional survey data. From a cost perspective, the prophylactic approach was riskier, because when cereal leafbeetle densities were high, economic loss was also high. However, fields under the prophylactic approach did not exceed threshold as often as fields using IPM. Total cost of prophylactic management was also $20.72 less per hectare, giving this approach an economic advantage over IPM. The majority of fields under the IPM approach did not exceed the economic threshold. Hence, from an economic perspective, both the prophylactic and IPM approaches have advantages and disadvantages. This helps explains the partial, rather than complete, adoption of IPM by southeastern U.S. wheat growers. Cereal leaf beetle was spatially aggregated across the region in 2010, but not in 2011. As a result, from an economic standpoint, prophylaxis or IPM may have a better fit in localized areas of the region than others. Finally, because IPM adoption is favored when it has a strong economic advantage over alternative management approaches, more emphasis should be placed on research to reduce costs within the IPM approach.

Relationship between external stink bug (Hemiptera: Pentatomidae) boll-feeding symptoms and internal boll damage with respect to cotton lint gin-out and fiber quality.

Cotton, Gossypium hirsutum L., bolls from 17 field locations in northeastern North Carolina and southeastern Virginia, having 20% or greater internal boll damage, were studied to determine the relationship between external feeding symptoms and internal damage caused by stink bug (Hemiptera: Pentatomidae) feeding. In 2006 and 2007, two cohorts of 100 bolls each were sampled at all field locations. The first cohort was removed as bolls reached approximately quarter size in diameter (2.4 cm). External and internal symptoms of stink bug feeding were assessed and tabulated. Concurrent to when the first cohort was collected, a second cohort of quarter-size-diameter bolls was identified, tagged, examined in situ for external feeding symptoms (sunken lesions), and harvested at the black seed coat stage. Harvested bolls were assessed for internal damage and locks were categorized (undamaged, minor damage, or major damage), dried, and ginned. Lint samples from each damage category were submitted for high volume instrument and advanced fiber information system quality analyses. Significant, moderately strong Pearson correlation coefficients existed between number of external stink bug feeding lesions and internal damage. Pearson correlation of total external lesions with total internal damage was stronger than any correlation among the other single components compared. Predictability plots indicated a rapid increase in relationship strength when relating external stink bug lesions to internal damage as the number of external lesions increased. Approximately 90% predictability of internal damage was achieved with four (2006) or six (2007) external lesions per boll. Gin-turnout and fiber quality decreased with increasing intensity of internal stink bug damage.

Fidelity of external boll feeding lesions to internal damage for assessing stink bug damage in cotton.

Previous research showed that the most precise estimates of stink bug damage in developing cotton bolls are obtained by collecting soft quarter-sized bolls and dissecting them for signs of internal feeding damage, such as internal boll wall warts and/or stained lint. However, this method requires considerable time and effort; therefore, scouts and growers are unwilling to invest adequate resources to make sound pest management decisions. Here, the authors evaluated enumeration of external feeding lesions on groups of 10, 15, 20, or 25 bolls per sample as an alternative sampling procedure. Results relate the similarity of external boll feeding lesions to internal damage as a function of boll sample size and external lesion tally. Inverse prediction confidence intervals (CIs) were also calculated to predict internal boll damage on a new sample based only on external feeding lesions. Results show that linear regression model fit increased when examining at least 20 bolls per sample, and only one external lesion per boll provided as good of model fit as using a minimum of two, three, or four lesions per boll. Inverse prediction CIs suggested that more than one sample (20 bolls per sample) will be required to make external lesions an acceptable method for making reasonably accurate management decisions. F-test lack of fit and significant regression models suggest that examination of external lesions is a promising a method for estimating stink bug damage in cotton.

Potential for Nezara viridula (Hemiptera: Pentatomidae) to transmit bacterial and fungal pathogens into cotton bolls.

Recently, we showed that the southern green stink bug (SGSB), Nezara viridula (L.), can transmit Pantoea agglomerans (Ewing and Fife), an opportunistic bacterium, into green cotton bolls resulting in plant disease. Here, we hypothesized that our established model could be used to determine if the SGSB was a general, non-discriminate vector by using two other opportunistic bacterial pathogens of bolls (Pantoea ananatis [Serano] and Klebsiella pneumoniae [Schroeter]) and the known fungal pathogen Nematospora coryli (Peglion). Variants of P. ananatis (strain Pa-1R) and K. pneumoniae (strain Kp 5-1R) selected for rifampicin (Rif) resistance were used as bacterial opportunists. N. coryli was detected only from laboratory-reared SGSB directly exposed to the fungus. Both Pa-1R and Kp 5-1R were recovered from SGSB previously provided a contaminated food source (2 days), sterile food (5 days), and then harvested after being caged on bolls (2 days) at levels reaching 10(3) and 10(4) colony forming units (cfus) per insect, respectively. However, bolls caged with insects infected with Pa-1R or Kp 5-1R and with evidence of feeding did not become diseased nor were either opportunists detected from boll tissues. Insects infected with N. coryli transmitted the pathogen, which resulted in diseased bolls and fungi concentrations reached 10(6) cfus/g locule tissue at 2 weeks following the caging period. Notably, each of the three pathogens independently caused boll disease when mechanically inoculated using a needle puncture. Generally, these results suggest that cotton pathogen acquisition by the SGSB was not sufficient to determine whether the insects were disease vectors of the opportunists.