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At least 91 records · Page 5

Overexpression of a plasma membrane protein generated broad-spectrum immunity in soybean

Plants fight-off pathogens and pests by manifesting an array of defence responses using their innate immunity mechanisms. Here we report the identification of a novel soybean gene encoding a plasma membrane protein, transcription of which is suppressed following infection with the fungal pathogen, Fusarium virguliforme. Overexpression of the protein led to enhanced resistance against not only against F. virguliforme, but also against spider mites (Tetranychus urticae, Koch), soybean aphids (Aphis glycines, Matsumura) and soybean cyst nematode (Heterodera glycines). We, therefore, name this protein as Glycine max disease resistance 1 (GmDR1; Glyma.10g094800). The homologues of GmDR1 have been detected only in legumes, cocoa, jute and cotton. The deduced GmDR1 protein contains 73 amino acids. GmDR1 is predicted to contain an ecto- and two transmembrane domains. Transient expression of the green fluorescent protein fused GmDR1 protein in soybean leaves showed that it is a plasma membrane protein. We investigated if chitin, a pathogen-associated molecular pattern (PAMP), common to all pathogen and pests considered in this study, can significantly enhance defence pathways among the GmDR1-overexpressed transgenic soybean lines. Chitin induces marker genes of the salicylic- and jasmonic acid-mediated defence pathways, but suppresses the defence pathway regulated by ethylene. Chitin induced SA- and JA-regulated defence pathways may be one of the mechanisms involved in generating broad-spectrum resistance among the GmDR1-overexpressed transgenic soybean lines against two serious pathogens and two pests including spider mites, against which no known resistance genes have been identified in soybean and among the most other crop species.

plasma membrane↗

α-Copaene is an attractant, synergistic with quercivorol, for improved detection of Euwallacea nr. fornicatus (Coleoptera: Curculionidae: Scolytinae)

The tea shot-hole borer, Euwallacea fornicatus Eichhoff, is an ambrosia beetle endemic to Asia and a pest of commercial tea, Camellia sinensis (L.) Kuntze. Recently, a complex of species morphologically similar to E. fornicatus has been recognized, which includes new pests established in Israel and the USA, both in California and Florida. Collectively termed E. nr. fornicatus, these cryptic species carry symbiotic Fusarium spp. fungi, some of which cause dieback disease in susceptible hosts, which include avocado, Persea americana Miller. Due to the threat to this economically important crop, research was initiated to evaluate efficacy of kairomone-based lures for detection of the beetle in Florida (termed the Florida tea shot hole borer, FL-TSHB). A series of field tests were conducted in 2016 in commercial avocado groves known to have FL-TSHB at various population levels. All tests evaluated lures containing quercivorol (p-menth-2-en-1-ol) andα-copaene, presented separately and in combination; and one test evaluated effect of trap type on beetle captures. In addition, electroantennography (EAG) was used to quantify female olfactory responses to lure emissions. This study identified (-)-α-copaene as a new attractant for FL-TSHB, equivalent in efficacy to quercivorol (the standard lure for Euwallacea detection in the USA); however, the combination of lures captured significantly more FL-TSHB than either lure alone. This combination resulted in synergistic attraction at two field sites and additive attraction at a third site. Sticky panel traps captured more FL-TSHB than comparably-baited Lindgren funnel traps. Females engaged in host-seeking flight from 11:00 to 16:00 hr (EST), with peak numbers observed between 12:00 and 13:00 hr. EAG analyses confirmed olfactory chemoreception of both kairomones, with a higher response elicited with the combination of volatiles. Results indicate that detection of pest E. nr. fornicatus in Florida can be improved by using a two-component lure consisting of p-menth-2-en-1-ol and (-)-α-copaene.

59 BASIC BIOLOGICAL SCIENCES↗

Calibration and Uncertainty Estimation Using the Ensemble Kalman Filter with a Large Subsurface Flow and Transport Model - 20321

At routinely monitored groundwater contamination sites, periodically measured environmental conditions such as groundwater levels and contaminant concentrations are used to inform and confirm a conceptual site model (CSM) and guide the development and calibration of a numerical groundwater flow and transport model. The calibration of groundwater flow and transport models after each measurement (sampling) event can illuminate deficiencies in a CSM, identify areas where additional monitoring is warranted, and predict the behavior of the system to guide decision making. However, manual and automated (e.g. PEST) model calibration tools can be time-consuming and computationally expensive to implement after each sampling event. Perhaps as a result, such calibration tools generally utilize all available monitoring data simultaneously rather than sequentially assimilating monitoring data one sampling event at a time as the results from sampling become available. A more real-time data assimilation approach may reduce parameter uncertainty, quantify the value of additional monitoring data, and produce a usable model more quickly and with less effort. To mitigate the potential time-consuming aspects of manual and widely applied automated calibration techniques, a data assimilation algorithm called the ensemble Kalman filter (EnKF) was evaluated as a relatively efficient method of model calibration and uncertainty assessment via the sequential integration of monitoring data into a model. The EnKF was able to successfully and efficiently assimilate monitoring and modeling data to calibrate a complex flow and transport model at a real-world site with significant subsurface heterogeneity, uncertainty, and 12 years of monitoring data (over 4,000 individual measurements of groundwater levels and over 2,500 measurements of contaminant concentrations). Starting with an uncalibrated model data from annual sampling events were sequentially assimilated, and the resultant predication errors and estimated parameter uncertainties were tracked. After all monitoring data were assimilated, both flow and transport residuals at the end of the EnKF process were comparable to those produced via a concurrent PEST calibration effort but required fewer model simulations. Both uncertainty and prediction errors decreased over time. In a real-time application, the adequacy of the model could be assessed after each sampling event. The benefits of such a real-time approach to utilizing monitoring data include reduced costs (in the form of model updates or site characterization efforts), early flagging of possible errors in the CSM, and a reduced risk of overfitting and corresponding increased confidence in model predictions. This tool may be particularly useful compared to other calibration techniques (e.g. manual, PEST) when model runtimes are long, calibration parameters are many, or parameter uncertainty is large. (authors)

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

MODFLOW6 models used to evaluate potential stresses and hydrologic conditions driving water-level fluctuations in well ER-5-3-2, Frenchman Flat, Southern Nevada

The hydrograph for well ER-5-3-2 in Frenchman Flat, southern Nevada, has previously unexplained water-level fluctuations. Four, three-dimensional, groundwater models (MODFLOW 6) were developed to evaluate potential stresses and hydrologic conditions affecting the well ER-5-3-2 hydrograph. Four model scenarios were developed that simulated: (1) wellbore leakage without recharge, (2) wellbore leakage with recharge, (3) shallow (low transmissivity) and deep (high transmissivity) carbonate rocks, and (4) lateral heterogeneity of carbonate rocks. Input and output files for the four model scenarios are in the model and output directories, respectively. Hydraulic conductivity, specific storage, and wellbore-leakage rates (when simulated) were estimated with parameter estimation (PEST) by minimizing a weighted composite, sum-of-squares objective function. The objective function was informed by measurement and Tikhonov regularization observations. Measurement observations included drawdowns from the constant-rate aquifer test and water-level altitudes measured in well ER-5-3-2 from 2001-2021. Tikhonov regularization informed hydraulic conductivity and specific storage parameters that were insensitive to measurement observations, where homogeneity was the preferred relation. Batch files, executables, and MODFLOW 6, PEST, and post-processing utilities are in the ancillary directory. Supplementary data also are included in the ancillary directory, including site information, high-frequency water-level and aquifer-test data, transmissivity estimates, water-chemistry data, and water-temperature analyses. This USGS data release contains data, analyses, and model files for the simulations and analysis results described in U.S. Geological Survey Scientific Investigations Report (https://doi.org/10.3133/sir20225132).

54 ENVIRONMENTAL SCIENCES↗

Climate-Induced Northerly Expansion of Siberian Silkmoth Range

Siberian silkmoth (Dendrolimus sibiricus Tschetv.) is a dangerous pest that has affected nearly 2.5 x 10(exp 6) ha of “dark taiga” stands (composed of Abies sibirica, Pinus sibirica and Picea obovata) within the latitude range of 52°–59° N. Here we describe a current silkmoth outbreak that is occurring about half degree northward of its formerly documented outbreak range. This outbreak has covered an area of about 800 thousand ha with mortality of conifer stands within an area of about 300 thousand ha. The primary outbreak originated in the year 2014 within stands located on gentle relatively dry southwest slopes at elevations up to 200 m above sea level (a.s.l.) Then the outbreak spread to the mesic areas including northern slopes and the low-elevation forest belts along the Yenisei ridge. Within the outbreak area, the northern Siberian silkmoth population has reduced generation length from two to one year. Our study showed that the outbreak was promoted by droughts in prior years, an increase of the sum of daily temperatures (t > +10 °C), and a decrease in ground cover moisture. Within the outbreak area, secondary pests were also active, including the aggressive Polygraphus proximus bark borer beetle. The outbreak considered here is part of the wide-spread (panzonal) Siberian silkmoth outbreak that originated during 2014–2015 with a range of up to 1000 km in southern Siberia. Our work concludes that observed climate warming opens opportunities for Siberian silkmoth migration into historically outbreak free northern “dark taiga” stands.

Siberian taiga↗

Genetic mapping of sugarcane aphid resistance in sorghum line SC112-14

Sugarcane aphid [Melanaphis sacchari (Zehntner)] is a destructive pest that has had an economic effect on sorghum in North America since 2013. The identification, development, and use of resistant sorghum germplasm is the most feasible strategy to control the pest. Nevertheless, the genetic control of sugarcane aphid (SCA) resistance is unknown for most sorghum resistant lines. To identify the genetic regions that confer SCA resistance in sorghum line SC112-14, 103 recombinant inbred lines (RILs) derived by its cross with the susceptible line PI 609251 were evaluated for their SCA resistance response in Georgia during two consecutive years. The resistance response was determined based on two ratings (2 wk apart) for aphid population size (APS) and aphid-induced plant damage (APD) each year. Segregation for SCA resistance was observed for the first APS and both APD ratings, and the broad-sense heritability estimate ranged from .71 to .76, respectively. A quantitative trait locus analysis using a high-density linkage map of 3,852 single nucleotide polymorphisms (SNPs) detected an 81-kb genomic region on chromosome 6 that explained 50–55% of the phenotypic variation. Comparative mapping analysis found that the resistance locus in SC112-14 is located 8- and 10-cM upstream of the Henong 16 (RMES1) and Tx2783 resistance loci, respectively, and encloses the SNP Sbv3.1_06_2316351 associated in Haitian resistant lines. Therefore, the line SC112-14 is an additional SCA resistance source that can be combined or strategically used with other resistance sources to assure a more robust host plant resistance to the SCA.

60 APPLIED LIFE SCIENCES↗

Resist! : Sustaining forest carbon sequestration and wood production after insect disturbance

Disturbances from insect pests threaten ecologically and economically important goods and services supplied by forests, including wood production and carbon sequestration. We highlight the factors that influence these services’ resistance, a term quantifying the initial response to disturbance. Insects inflict damage through a range of mechanisms, prompting distinct plant physiological responses that scale to influence ecosystem processes and, with time, goods and services. The degree and timing of tree mortality and defoliation affect the amount of residual vegetation available to support compensatory wood production and influence carbon sequestration by changing rates of detritus-fueled decomposition. Compounding, or sequential, insect attacks may prime a forest for additional disturbance, further eroding wood production and carbon sequestration. Forest management practices that promote biological and structural diversity, and augment or retain limiting biological and nutrient resources, may buffer against the effects of insect pests on wood production and carbon sequestration.

Gough, Christopher M. [Virginia Commonwealth Univ.↗

The binding properties of cycloxaprid on insect native nAChRs partially explain the low cross‐resistance with imidacloprid in Nilaparvata lugens

Abstract BACKGROUND Neonicotinoids, such as imidacloprid, are selective agonists of insect nicotinic acetylcholine receptors (nAChRs) used to control Nilaparvata lugens , a major rice insect pest. High imidacloprid resistance has been reported in N. lugens both in the laboratory and in the field. Cycloxaprid (CYC), an oxa‐bridged cis ‐nitromethylene neonicotinoid, showed high insecticidal activity against N. lugens and low cross‐resistance in imidacloprid‐resistant strains and field populations. RESULTS Binding studies demonstrated that imidacloprid has two binding sites with different affinities ( K d = 3.18 ± 0.43 p m and 1.78 ± 0.19 n m ) in N. lugens nAChRs. CYC was poor at displacing [ 3 H]imidacloprid at its high‐affinity binding site ( K i = 159.38 ± 20.43 n m ), but quite efficient at the low‐affinity binding site ( K i = 1.27 ± 0.35 n m ). These data showed that CYC had overlapping binding sites with imidacloprid only at its low‐affinity binding site. Therefore, the low displacement ability of CYC against imidacloprid binding at its high‐affinity site could partially explain the low cross‐resistance of CYC in imidacloprid‐resistant populations. CONCLUSION The high insecticidal activity, low cross‐resistance and different binding properties on insect nAChRs of CYC show that it is a potential insecticide for the control of N. lugens and related insect pests, especially ones with high resistance to neonicotinoids. © 2018 Society of Chemical Industry

Zhang, Yixi↗

The synergistic effects of rosehip oil and matrine against Icerya aegyptiaca (Douglas) (Hemiptera: Coccoidea) and the underlying mechanisms

Abstract BACKGROUND Icerya aegyptiaca (Douglas) is an important agricultural pest that has a protective wax shell. Insecticides generally fail to achieve the desired control of I. aegyptiaca because of difficulties penetrating this wax shell. Plant essential oils are rich in terpenoids and have strong penetrability and expansibility, which can enable the rapid penetration and absorption of insecticides, thereby improving the control effect. Matrine is a botanical insecticide with contact and stomach toxicity, low toxicity toward non‐target organisms and is environmentally friendly. In this study, we tested the insecticidal activity of rosehip oil (Ro)/matrine combinations and revealed the synergistic mechanism of Ro and its components with matrine in terms of physiology and biochemistry. RESULTS Ro/matrine combinations have strong penetrating power, enabling matrine to quickly penetrate the wax shell of I. aegyptiaca and enter the insect body. This improves the insecticidal activity and enhances inhibition of acetylcholinesterase activity. Ro and its main chemical constituents, cineole and (+)‐camphor, showed synergistic effects on matrine with synergic ratios of 4.79, 3.49 and 4.21, respectively. CONCLUSION Combinations of Ro, cineole and (+)‐camphor with matrine have good insecticidal effects on I. aegyptiaca while remaining safe to the environment. These combinations of biological insecticides have excellent development prospects and provide a new reference for the pest management of scale insects. © 2022 Society of Chemical Industry.

Zhou, Yi↗

Effects of trunk injection with emamectin benzoate on arthropod diversity

Abstract BACKGROUND Pine wood nematode is a major plant quarantine object in the world. Trunk injection is an effective method for controlling pests that cause disease. To evaluate the ecological safety of trunk injection with emamectin benzoates in forests of Pinus massoniana , the community diversity and community composition of soil arthropods and flying insects (Hymenoptera) were studied at different stages of trunk injection. RESULTS The dominant taxonomic groups of soil arthropods were Collembola (30.80%), Insecta (26.42%), and Arachnida (23.84%). The taxonomic groups of flying insects (Hymenoptera) were Ichneumonidae (48.94%), Formicidae (14.10%), and Braconidae (8.44%). Trunk injection with emamectin benzoate has no significant effect on the community diversity indices of total soil arthropods and flying insects (Hymenoptera). However, it has a significant effect on the community diversity indices of detritivores for soil arthropods. It changed the community composition of soil arthropods but did not impact the community composition of flying insects (Hymenoptera). Redundancy analysis of arthropod community structure and environmental variables showed that total potassium, residual of green leaf, and residual of litter leaf have a significant impact on the community structure of soil arthropods, and total phosphorus, total nitrogen, water content, organic matter, and total potassium have a significant impact on the community structure of flying insects (Hymenoptera). CONCLUSION Trunk injection with emamectin benzoate is safe for the ecological environment. This study provides a new insight into the field for the prevention and control of pine wood nematode disease, which is of great significance to forest management and pest control. © 2022 Society of Chemical Industry.

Ouyang, Xianheng↗

Role of Zr in strengthening MoSi 2 from density functional theory calculations

MoSi 2 is an important intermetallic with excellent oxidation resistance at high temperatures above1000C. However, its application at lower temperatures is limited by oxygen embrittlement, a phenomenon known as “pesting”. In this work, we comprehensively investigate the role of Zr in mitigating pesting in MoSi2using density functional theory calculations. We show that Zr dopants reduce the embrittling effects of oxygen interstitials at MoSi 2 grain boundaries by being a charge donor to oxygen. However, a more substantial effect is observed when Zr is present as a secondary getter nanoparticle phase. Oxygen interstitials have a strong thermodynamic driving force to migrate into the Zr subsurface at the Zr/MoSi 2 interface, and the work of separation of the clean and oxygen-contaminated Zr/MoSi 2 interfaces are much higher than that of MoSi 2 grain boundaries. So we present an efficient screening approach to identify other potential getter elements using simple thermodynamic descriptors, which can be extended to other alloy systems of interest. These findings provide crucial fundamental insight s and further avenues to optimize Mo and other alloys.

36 MATERIALS SCIENCE↗

An algal-bacterial symbiotic system of carbon fixation using formate as a carbon source

The commercial implementation of algal cultures for efficient capture of CO 2 from fossil-derived flue gases is not a reality yet due to several major challenges, particularly low gas-liquid mass transport efficiency along with loss of productivity due to pests. This study has explored an algal-bacterial symbiotic system to utilize formate as a carbon source. The algal-bacterial assemblage, after a 400-day adaptive evolution using the formate medium, has demonstrated a new capability to assimilate both formate and CO 2 to promote biomass production. The formate tolerant culture system also addresses CO 2 mass transfer limitation under high light conditions. Continuous cultivation of the assemblage on formate led to a carbon capture efficiency of 90 % with a biomass concentration of 0.92 g L -1 and biomass productivity of 0.31 g L -1 day -1 , and the system did not require CO 2 aeration. In addition, 13 C isotope tracing, proteomics, and microbial community analysis reveal a uniquely evolved community for formate utilizations and robust biomass synthesis, in which Chlorella sorokiniana is a dominant phototroph companying with highly diverse bacterial subpopulations. In the presence of formate, evidence has shown that Chlorella is able to form mutualism with formate utilizing bacteria in semi-continuous cultures, leading to a highly effective photomixotrophic metabolism. Furthermore, this study demonstrates a new route of using electrochemical-derived formate to support mutualistic algae-bacteria biorefinery while the formate as an alternative carbon source could repel pests for outdoor algal cultivations.

59 BASIC BIOLOGICAL SCIENCES↗

Predicting plant attractiveness to coccinellids with plant trait profiling, citizen science, and common garden surveys

Lady beetles are key predators in agricultural systems and their impact can be enhanced by plant resources. Facilitation of lady beetles is a potentially effective pest management strategy but a lack of data on which plant species attract and nurture them limits implementation. Here we investigate which plant species might optimize lady beetle impact by developing a profile of plant characteristics most likely to attract lady beetles and testing if the subset of “potentially preferred” plant families exhibiting those characteristics are significantly more attractive to lady beetles than other families. This testing encompassed a combination of observations submitted to a citizen science program, the Lost Ladybug Project, and two repeated site surveys. Traits reported to be most attractive to lady beetles included the presence of trichomes that provide protection from predation, nutritional resources such as nectar or prey, and visual or chemical cues associated with those traits. Significantly more lady beetles were observed on the three plant families which most often exhibit these traits, Apiaceae, Asteraceae, and Rosaceae, than on other families. Furthermore, strategies for incorporating plants from these families into production systems to attract lady beetles and limit pest damage are discussed.

59 BASIC BIOLOGICAL SCIENCES↗

Solving high-dimensional inverse problems using amortized likelihood-free inference with noisy and incomplete data

Here, we present a likelihood-free probabilistic inversion method based on normalizing flows for high-dimensional inverse problems. The proposed method is composed of two complementary networks: a summary network for data compression and an inference network for parameter estimation. The summary network encodes raw observations into a fixed-size vector of summary features, while the inference network generates samples of the approximate posterior distribution of the model parameters based on these summary features. The posterior samples are produced in a deep generative fashion by sampling from a latent Gaussian distribution and passing these samples through an invertible transformation. We construct this invertible transformation by sequentially alternating conditional invertible neural network and conditional neural spline flow layers. The summary and inference networks are trained simultaneously. We apply the proposed method to an inversion problem in groundwater hydrology to estimate the posterior distribution of the log-conductivity field conditioned on spatially sparse time-series observations of the system’s hydraulic head responses. The conductivity field is represented with 706 degrees of freedom in the considered problem. Comparison with the likelihood-based iterative ensemble smoother PEST-IES method demonstrates that the proposed method accurately estimates the parameter posterior distribution and the observations’ predictive posterior distribution at a fraction of the inference time of PEST-IES.

conditional invertible neural network↗

A numerical study of process complexity in permafrost dominated regions

Numerical modeling of permafrost dynamics requires adequate representation of atmospheric and surface processes, a reasonable parameter estimation strategy, and site-specific model development. The three main research objectives of the study are: (i) to propose a novel methodology that determines the required level of surface process complexity of permafrost models by conducting parameter sensitivity and calibration, (ii) to design and compare three numerical models of increasing surface process complexity, and (iii) to calibrate and validate the numerical models at the Yakou catchment on the Qinghai-Tibet Plateau as an exemplary study site. The calibration was carried out by coupling the Advanced Terrestrial Simulator (numerical model) and PEST (calibration tool). Simulation results showed that (i) A simple numerical model that considers only subsurface processes can simulate active layer development with the same accuracy as other more complex models that include surface processes. (ii) Peat and mineral soil layer permeability, Van Genuchten alpha, and porosity are highly sensitive. (iii) Liquid precipitation aids in increasing the rate of permafrost degradation. (iv) Deposition of snow insulated the subsurface during the thaw initiation period. We have developed and released an integrated code that couples the numerical software ATS to the calibration software PEST. The numerical model can be further used to determine the impacts of climate change on permafrost degradation.

Calibration↗

The movement of small insects in the convective boundary layer: linking patterns to processes

In fine warm weather, the daytime convective atmosphere over land areas is full of small migrant insects, among them serious pests (e.g. some species of aphid), but also many beneficial species (e.g. natural enemies of pests). For many years intensive aerial trapping studies were the only way of determining the density profiles of these small insects, and for taxon-specific studies trapping is still necessary. However, if we wish to determine generic behavioural responses to air movements shown by small day-migrating insects as a whole, the combination of millimetre-wavelength ‘cloud radars’ and Doppler lidar now provides virtually ideal instrumentation. Here we examine the net vertical velocities of >1 million insect targets, relative to the vertical motion of the air in which they are flying, as a succession of fair-weather convective cells pass over the recording site in Oklahoma, USA. The resulting velocity measurements are interpreted in terms of the flight behaviours of small insects. These behaviours are accounted for by a newly-developed Lagrangian stochastic model of weakly-flying insect movements in the convective boundary layer; a model which is consistent with classic characterisations of small insect aerial density profiles. We thereby link patterns to processes.

54 ENVIRONMENTAL SCIENCES↗

Linking Small-Scale Flight Manoeuvers and Density Profiles to the Vertical Movement of Insects in the Nocturnal Stable Boundary Layer

Huge numbers of insects migrate over considerable distances in the stably-stratified night-time atmosphere with great consequences for ecological processes, biodiversity, ecosystem services and pest management. We used a combination of meteorological radar and lidar instrumentation at a site in Oklahoma, USA, to take a new look at the general assistance migrants receive from both vertical and horizontal airstreams during their long-distance flights. Movement in the nocturnal boundary layer (NBL) presents very different challenges for migrants compared to those prevailing in the daytime convective boundary layer, but we found that Lagrangian stochastic modelling is effective at predicting flight manoeuvers in both cases. A key feature for insect transport in the NBL is the frequent formation of a thin layer of fast-moving air – the low-level jet. Modelling suggests that insects can react rapidly to counteract vertical air movements and this mechanism explains how migrants are retained in the jet for long periods (e.g. overnight, and perhaps for several hours early in the morning). This results in movements over much longer distances than are likely in convective conditions, and is particularly significant for the reintroduction of pests to northern regions where they are seasonally absent due to low winter temperatures.

54 ENVIRONMENTAL SCIENCES↗

Knockout of ABC transporter gene ABCA2 confers resistance to Bt toxin Cry2Ab in Helicoverpa zea

Evolution of pest resistance reduces the benefits of widely cultivated genetically engineered crops that produce insecticidal proteins derived from Bacillus thuringiensis (Bt). Better understanding of the genetic basis of pest resistance to Bt crops is needed to monitor, manage, and counter resistance. Previous work shows that in several lepidopterans, resistance to Bt toxin Cry2Ab is associated with mutations in the gene encoding the ATP-binding cassette protein ABCA2. The results here show that mutations introduced by CRISPR/Cas9 gene editing in the Helicoverpa zea (corn earworm or bollworm) gene encoding ABCA2 (HzABCA2) can cause resistance to Cry2Ab. Disruptive mutations in HzABCA2 facilitated the creation of two Cry2Ab-resistant strains. A multiple concentration bioassay with one of these strains revealed it had > 200-fold resistance to Cry2Ab relative to its parental susceptible strain. All Cry2Ab-resistant individuals tested had disruptive mutations in HzABCA2. We identified five disruptive mutations in HzABCA2 gDNA. The most common mutation was a 4-bp deletion in the expected Cas9 guide RNA target site. The results here indicate that HzABCA2 is a leading candidate for monitoring Cry2Ab resistance in field populations of H. zea.

60 APPLIED LIFE SCIENCES↗