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Prior to the recent implementation of the Mpp51Aa2 pesticidal protein (ThryvOn), transgenic cotton cultivars have historically offered no control of the cotton fleahopper ( Pseudatomocelis seriatus (Reuter)). To evaluate the feeding behavior of cotton fleahoppers on ThryvOn cotton, electropenetrography (EPG) using a Giga-8 DC instrument was used to monitor the probing activity of fourth- and fifth-instar cotton fleahopper nymphs on both ThryvOn and non-ThryvOn cotton squares. Nymphs were individually placed on an excised cotton square for 8 h of EPG recording, after which resulting waveforms were classified as non-probing, cell rupturing, or ingestion. Although there were significantly more cell rupturing events per insect on ThryvOn (mean ± SEM, 14.8 ± 1.7) than on non-ThryvOn squares (mean ± SEM, 10.3 ± 1.6), there was no difference attributable to ThryvOn in the average number of ingestion events per insect. However, the average duration of ingestion events was significantly shorter on squares with ThryvOn (mean ± SEM, 509 ± 148 s) than on squares without (mean ± SEM, 914 ± 135 s). This suggests that cotton fleahoppers continued to probe despite their inability to sustain ingestion. These results provide conclusive evidence that the Mpp51Aa2 pesticidal protein affects the feeding behavior of cotton fleahopper nymphs.

Extensive genome structure variations, such as copy number variations (CNVs) and presence/absence variations, are the basis for the remarkable genetic diversity of maize; however, the effect of CNVs on maize herbivory defense remains largely underexplored. Here, we report that the naturally occurring duplication of the maize 9-lipoxygenase gene ZmLOX5 leads to increased resistance of maize to herbivory by fall armyworms (FAWs). Previously, we showed that ZmLOX5-derived oxylipins are required for defense against chewing insect herbivores and identified several inbred lines, including Yu796, that contained duplicated CNVs of ZmLOX5 , referred to as Yu796-2×LOX5 . To test whether introgression of the Yu796-2×LOX5 locus into a herbivore-susceptible B73 background that contains a single ZmLOX5 gene is a feasible approach to increase resistance, we generated a series of near-isogenic lines that contained either two, one, or zero copies of the Yu796-2×LOX5 locus in the B73 background via six backcrosses (BC6). Droplet digital PCR (ddPCR) confirmed the successful introgression of the Yu796-2×LOX5 locus in B73. The resulting B73-2× LOX5 inbred line displayed increased resistance against FAW, associated with increased expression of ZmLOX5 , increased wound-induced production of its primary oxylipin product, the α-ketol, 9-hydroxy-10-oxo-12(Z),15(Z)-octadecadienoic acid (9,10-KODA), and the downstream defense hormones regulated by this molecule, 12-oxo-phytodienoic acid (12-OPDA) and abscisic acid (ABA). Surprisingly, wound-induced JA-Ile production was not increased in B73-2× LOX5 , resulting from the increased JA catabolism. Furthermore, B73-2× LOX5 displayed reduced water loss in response to drought stress, likely due to increased ABA and 12-OPDA content. Taken together, this study revealed that the duplicated CNV of ZmLOX5 quantitively contributes to maize antiherbivore defense and presents proof-of-concept evidence that the introgression of naturally occurring duplicated CNVs of a defensive gene into productive but susceptible crop varieties is a feasible breeding approach for enhancing plant resistance to herbivory and tolerance to abiotic stress.

Fusarium wilt of cotton caused by the soilborne fungal pathogen Fusarium oxysporum f. sp. vasinfectum race 4 (FOV4) is a contemporary epidemic affecting cotton production in Far West Texas. The spatial distribution of soilborne FOV4 can be heterogeneous at small scales, and the factors that lead to this heterogeneity require investigation. Hypothetical causes include dissemination of spores through soils and variable saprophytic growth of the fungus. In the field, FOV4 DNA was quantified from soil during and after the cotton-growing season, and though the average amounts of DNA were not different between these time points, the variances of DNA across space were significantly different. Variability was higher when pathogenic growth of the fungus was expected owing to the presence of live cotton plants and lower when saprophytic growth was expected after cropping. In sterile-environment growth chamber experiments, the abundance of organic matter influenced the fungal vegetative growth rate and maximum amount as measured through quantitative PCR and the timing of the fungus' increasing its rate of spore production as measured through dilution plating. To investigate movement of spores in soils, spore mobility in experimental columns was quantified. Soil composition and organic matter abundance affected spore mobility, indicating that the timing of spore production relative to the availability of growth resources will affect the spatial spread of FOV4 and suggesting that soil properties affect the retention of conidia. The spatial spread of FOV4 through soil varies temporally and is affected by the shift between pathogenic and saprophytic growth modes of the fungus., Competing Interests: The author(s) declare no conflict of interest.

Auxotrophic strains of Agrobacterium tumefaciens can contribute to the development of more efficient transformation systems, especially for crops historically considered recalcitrant. Homologous recombination was used to derive methionine auxotrophs of two common A. tumefaciens strains, LBA4404 and EHA105. The EHA105 strains were more efficient for switchgrass transformation, while both the EHA105 and LBA4404 strains worked equally well for the rice control. Event quality, as measured by transgene copy number, was not affected by auxotrophy, but was higher for the LBA4404 strains than the EHA105 strains. Ultimately, the use of auxotrophs reduced bacterial overgrowth during co-cultivation and decreased the need for antibiotics., (© 2022. The Author(s).)

Colletotrichum graminicola is an economically significant fungal pathogen of maize. The primary infective conidia of the fungus, falcate conidia, are splash-dispersed during rain events. The adhesion of the falcate conidia triggers germination and is required for the development of infection structures. Falcate conidia are capable of immediate adhesion upon encountering the substrate. We report that rapid adhesion in C. graminicola is polarized, with a single-sided strip of adhesive material running the length of a single side (or face) of the conidium between the tips. This strip of adhesive is co-localized with dynamic transverse actin cables, and both the adhesive strip and actin cables are formed after liberation of the conidium from its conidiogenous cell but prior to adhesion to the infection court. Orientation of conidia upon contact with substrate determines whether they will rapidly adhere, and those which do not initially adhere can be induced to do so by applying force to reorient or "flip" the conidia. We propose that C. graminicola possesses an adhesive mechanism resulting in an adhesion efficiency of approximately 50% upon initial contact with substrata, and that an increase in adhesion efficiency can be induced by disturbance., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Fusarium wilt of cotton, caused by the soilborne fungal pathogen Fusarium oxysporum f. sp. vasinfectum (FOV), occurs in regions of the United States where cotton ( Gossypium spp.) is grown. Race 4 of this pathogen (FOV4) is especially aggressive, and does not require the co-occurrence of the root knot nematode ( Meloidogyne incognita ) to infect cotton. Its sudden appearance in far-west Texas in 2016 after many years of being restricted to California is of great concern, as is the threat of its continued spread through the cotton-producing regions of the United States. The aim of this research was to analyze the spatial variability of FOV4 inoculum density in the location where FOV4 is locally emerging, using quantitative and droplet digital PCR methods. Soil samples collected from a field with known FOV4 incidence in Fabens, Texas, were analyzed. Appreciable variation in inoculum density was found to occur at spatial scales smaller than the size of plots involved in cultivar trial research, and was spatially autocorrelated (Moran's I, Z = 17.73, P < 0.0001). These findings indicate that, for cultivar trials, accounting for the spatial distribution of inoculum, either by directly quantifying it or through the use of densely distributed calibration checks, is important to the interpretation of results.

Background: Thrips (order Thysanoptera) infestations of cotton seedlings result in plant injury, increasing the detrimental consequences of other challenges to production agriculture, such as abiotic stress or infestation by other pests. Using Frankliniella fusca as a thrips species of focus, we empirically developed a composite model of thrips phenology and cotton seedling susceptibility to predict site-specific infestation risk so that monitoring and other resources can be allocated efficiently, to optimize the timing of thrips control measures to maximize effectiveness, and to inform stakeholders about the dynamics of thrips infestation and cotton seedling injury at a time when thrips are evolving resistance to commonly-used pesticides., Results: A mixture distribution model of thrips infestation potential, fit to data describing F. fusca adult dispersal in time, proved best for predicting infestations of F. fusca on cotton seedlings. Thrips generations occurring each year as a function of weather are represented as a probability distribution. A model of cotton seedling growth was also developed to predict susceptibility as a function of weather. Combining these two models resulted in a model of seedling injury, which was validated and developed for implementation as a software tool., Conclusions: Experimental validation of the implemented model demonstrated the utility of its output in predicting infestation risk. Successful implementation and use of the software tool derived from this model was enabled by close cooperation with university extension personnel, agricultural consultants, and growers, underscoring the importance of stakeholder and expert input to the success of applied analytical research. © 2020 Society of Chemical Industry., (© 2020 Society of Chemical Industry.)

Management of disease affecting peanut in the southeastern United States has benefited from extensive field research identifying disease-associated risk factors since the 1990s. An assessment of risk factors associated with tomato spotted wilt (TSW), caused by tomato spotted wilt virus and spread exclusively by thrips, is available to growers through Peanut Rx, a tool developed to inform peanut management decisions. Peanut Rx provides an assessment of relative TSW risk as an index. The assessment provides information about the relative degree to which a field characterized by a specified suite of practices is at risk of crop loss caused by TSW. Loss results when infection occurs, and infection rates are determined, in part, by factors outside a grower's control, primarily the abundance of dispersing, viruliferous thrips. In this study, we incorporated meteorological variables useful for predicting thrips dispersal, increasing the robustness of the Peanut Rx framework in relation to variation in the weather. We used data from field experiments and a large grower survey to estimate the relationships between weather and TSW risk mediated by thrips vectors, and developed an addition to Peanut Rx that proved informative and easy to implement. The expected temporal occurrence of major thrips flights, as a function of heat and precipitation, was translated into the existing risk-point system of Peanut Rx. Results from the grower survey further demonstrated the validity of Peanut Rx for guiding growers' decisions to minimize risk of TSW.

Agricultural industrialization and the subsequent reliance on pesticides has resulted in numerous unintended consequences, such as impacts upon the environment and by extension human health. Eco-efficiency is a strategy for sustainably increasing production, while simultaneously decreasing these externalities on ecological systems. Eco-efficiency is defined as the ratio of production to environmental impacts. It has been widely adopted to improve chemical production, but we investigate the challenges of applying eco-efficiency to pesticide use. Eco-efficiency strategies include technological innovation, investment in research and development, improvement of business processes, and accounting for market forces. These components are often part of integrated pest management (IPM) systems that include alternatives to pesticides, but its implementation is often thwarted by commercial realities and technical challenges. We propose the creation and adoption of an eco-efficiency index for pesticide use so that the broad benefits of eco-efficient strategies such as IPM can be more readily quantified. We propose an index based upon the ratio of crop yield to a risk quotient (RQ) calculated from pesticide toxicity. Eco-efficiency is an operational basis for optimizing pest management for sustainability. It naturally favors adoption of IPM and should be considered by regulators, researchers, and practitioners involved in pest management. © 2019 Society of Chemical Industry., (© 2019 Society of Chemical Industry.)

There is increasing evidence that acylsugars deter insect pests and plant virus vectors, including the western flower thrips (WFT), Frankliniella occidentalis (Pergande), vector of tomato spotted wilt virus (TSWV). Acylsugars are sugar-polyesters composed of saturated, un-saturated, and variously branched short and long chain fatty acids (FAs) esterified to a glucose (acylglucose) or sucrose (acylsucrose) moiety. We sought to understand how acylsucrose amount and composition of associated FA profiles interacted to mediate resistance to WFT oviposition and TSWV inoculation on tomato leaves. Towards this goal, we examined WFT oviposition and TSWV inoculation behavior on tomato lines bred to exude varying amounts of acylsucrose in association with diverse FA profiles. Our data show that as acylsucrose amounts increased, WFT egg-laying (oviposition) decreased and TSWV inoculation was suppressed. Western flower thrips also responded to FA profiles that included iC4, iC11, nC12 and nC10 FA. These findings support improving acylsugar-mediated resistance against WFT by breeding tomatoes exuding greater amounts of acylsucrose associated with specific FA profiles. We show that increasing acylsucrose amount output by type IV trichomes and selecting for particular FA profiles through advanced breeding profoundly affects WFT behavior in ways that benefit management of WFT as direct pests and as TSWV vectors.

Several problems limit the productivity and acceptance of crop protection, including pesticide overuse, pesticide resistance, poor adoption of integrated pest management (IPM), declining funding for research and extension, and inefficiencies of scale. We discuss the proposition that alternative business models for crop protection can address these problems by incentivizing and benefiting from efficiency of pesticide use. Currently, business models are not linked to the adoption of IPM and are sometimes at odds with IPM practices. We explore a business model based on the provision of pest management adequacy through services rather than the sale of pesticide products. Specifically, we advocate for establishment of crop protection adequacy standards that would allow a market system to maximize efficiency. Changing some of the relationships between agricultural companies and producers from one based on products to one based on services is an idea worthy of debate and evaluation for improving the efficiency of pest management. Contemporary information technology enhancing monitoring and coordination warrants attention in this debate. © 2019 Society of Chemical Industry., (© 2019 Society of Chemical Industry.)

Background: Limb length discrepancy (LLD) in the setting of concurrent hindfoot and ankle deformity poses an added level of complexity to the reconstructive surgeon. Regardless of etiology, a clinically significant LLD poses additional challenges without a forthright and validated solution. The purpose of the current study is to determine whether reconstructive hindfoot and ankle surgery with concurrent lengthening through a distal tibial corticotomy is comparable to other treatment alternatives in the literature., Patients and Methods: A retrospective review of hindfoot and ankle deformity correction utilizing Ilizarov circular external fixation with concurrent distal tibial distraction osteogenesis from July 2009 to September 2014 was conducted., Results: This study included 19 patients with a mean age of 47.47 ± 13.36 years with a mean follow up of 576.13 ± 341.89 days. The mean preoperative LLD was 2.70 ± 1.22 cm and the mean operatively induced LLD was 2.53 ± 0.59 cm. The mean latency period was 9.33 ± 3.47 days and distraction rate was 0.55 ± 0.16 mm/day. The mean distraction length was 2.14 ± 0.83 cm and mean duration of external fixation was 146.42 ± 58.69 days. The time to union of all hindfoot and ankle fusions was 121.00 ± 25.66 days with an overall fusion rate of 85.71%., Conclusions: The successful treatment of hindfoot and ankle deformity correction in the setting of LLD using the technique of a distal tibial corticotomy and distraction osteogenesis is reported and illustrates an additional treatment technique with comparable measured outcomes to those previously described. We urge that each patient presentation be evaluated with consideration of all described approaches and associated literature to determine the current best reconstructive approach as future studies may validate or replace the accepted options at present.

Background: Insecticide resistance arises at a given location in response to selection acting on novel genotypes or standing variation, or allelic migration. Fitness costs of resistance may slow resistance evolution or result in reversion to susceptibility, but consistent and geographically widespread use of insecticides may provide sufficient selection to offset the fitness costs of resistance. Understanding this relationship is important to the success of insecticide resistance management. We report the existence of fitness costs of neonicotinoid resistance in field-collected populations of the tobacco thrips (Frankliniella fusca), which increasingly challenge upland cotton production in the southeastern USA., Results: Populations (14 of 15 in 2015; 4 of 5 in 2016) investigated showed a loss of resistance to imidacloprid after multiple generations without exposure to the insecticide. Populations studied in 2016 were each split into two colonies, and one of each pair was repeatedly exposed to imidacloprid. In three of the four populations that lost resistance, imidacloprid-exposed colonies lost resistance significantly more slowly than did corresponding unexposed colonies., Conclusion: For imidacloprid resistance to be broadly increasing in the landscapes of the southeastern USA despite fitness costs of resistance, selection for resistance must be sufficient to overcome the costs. Findings encourage investigation into why costs are overcome in this system, potentially including geographic extent of neonicotinoid use or prevalence of low-dose exposure. © 2019 Society of Chemical Industry., (© 2019 Society of Chemical Industry.)

Imidacloprid is widely used to manage tomato spotted wilt disease (TSW) in tobacco, tomato, and pepper, caused by Tomato spotted wilt orthotospovirus (TSWV) and spread by the tobacco thrips, Frankliniella fusca Hinds (Thysanoptera: Thripidae). Imidacloprid suppresses transmission of TSWV by reducing probing and feeding by adult thrips on treated plants, thereby reducing the probability of transmission by infectious thrips. Because imidacloprid does not reduce probing and feeding on treated plants to zero, the reduction in transmission probability per viruliferous thrips can be offset by an increase in the number of viruliferous thrips challenging treated plants. A composite of these effects which we call 'pathogen pressure' experienced by plants is a function of thrips population size, the proportion of those thrips that are viruliferous, and the probability that viruliferous thrips successfully inoculate plants. To better understand the relationship between imidacloprid's effect on virus transmission, pathogen pressure, and TSW incidence in tobacco, we modeled TSW incidence as a function of the two most important variables affecting components of pathogen pressure, temperature, and precipitation, and the dependence of imidacloprid's effect on pathogen pressure. A model incorporating imidacloprid's effect as a reduction in pathogen pressure was found to be more descriptive than models incorporating the effect as a reduction in TSW incidence. Results reveal maximum proportional reduction in TSW incidence resulting from imidacloprid use is associated with minimal potential TSW incidence. As pathogen pressure increases, potential TSW incidence approaches 100%, and the benefits of imidacloprid use are highest at intermediate levels of pathogen pressure.

Tomatoes (Solanum lycopersicum L.) have been bred to exude higher amounts or different types of the specialized plant metabolites, acylsugars, from type IV trichomes. Acylsugars are known to deter several herbivorous insect pests, including the western flower thrips (WFT), Frankliniella occidentalis (Pergande); however, all previous studies investigated the effect of acylsugars on leaves, or acylsugar extracts obtained from leaves. In spite of the WFT predilection for flowers, there is a gap in knowledge about flower defenses against thrips damage. This is especially important in light of their capacity to acquire and inoculate viruses in the genus Orthotospovirus, such as Tomato spotted wilt orthotospovirus (TSWV), in flowers. Therefore, we turned our attention to assessing thrips oviposition differences on flowers of 14 entries, including 8 interspecific hybrids, 5 tomato lines bred for specific acylsugar-related characteristics (type IV trichome densities, acylsugar amount, sugar moiety and fatty acid profile), and a fresh market tomato hybrid, Mt. Spring, which only produces trace amounts of acylsugars. Our results show that the density of the acylsugar droplet bearing type IV trichomes is greatest on sepals, relative to other flower structures, and accordingly, WFT avoids oviposition on sepals in favor of trichome-sparse petals. In concordance with past studies, acylsugar amount was the most important acylsugar-related characteristic suppressing WFT oviposition. Certain acylsugar fatty acids, specifically i-C5, i-C9 and i-C11, were also significantly associated with changes in WFT oviposition. These results support continued breeding efforts to increase acylsugar amounts and explore modifications of fatty acid profile and their roles in deterring thrips oviposition. The finding that acylsugar production occurs and reduces thrips oviposition in tomato flowers will be important in efforts to use acylsugar-mediated resistance to reduce incidence of orthotospoviruses such as TSWV in tomato by deterring virus transmission and development of thrips vector populations in the crop., Competing Interests: The authors have declared that no competing interests exist.

[This corrects the article DOI: 10.1371/journal.pone.0073321.].

The intensification of industrial agriculture has been enabled by improved crop varieties, genetically engineered crops, fertilizers, and pesticides. Over the past 15 years, neonicotinoid seed treatments have been adopted worldwide and are used on a large proportion of U.S. field crops. Although neonicotinoids are used widely, little is known about how large-scale deployment affects pest populations over long periods. Here, we report a positive relationship between the deployment of neonicotinoid seed-dressings on multiple crops and the emergence of insecticide resistance in tobacco thrips (Frankliniella fusca), a polyphagous insect herbivore that is an important pest of seedling cotton but not soybean or maize. Using a geospatial approach, we studied the relationship between neonicotinoid resistance measured in 301 F. fusca populations to landscape-scale crop production patterns across nine states in the southeastern U.S. cotton production region, in which soybean, maize and cotton are the dominant crops. Our research linked the spatiotemporal abundance of cotton and soybean production to neonicotinoid resistance in F. fusca that is leading to a dramatic increase in insecticide use in cotton. Results demonstrate that cross-crop resistance selection has important effects on pests and, in turn, drives pesticide use and increases environmental impacts associated with their use.

Background: Over the past two decades, neonicotinoid seed treatments have become the primary method to manage tobacco thrips, Frankliniella fusca Hinds, on seedling cotton. Because this insect is highly polyphagous and the window of insecticide exposure is short, neonicotinoid resistance was expected to pose a minimal risk. However, reports of higher than expected F. fusca seedling damage in seed-treated cotton fields throughout the Mid-South and Southeast US production regions suggested neonicotinoid resistance had developed. To document this change, F. fusca populations from 86 different locations in the eastern United States were assayed in 2014 and 2015 for imidacloprid and thiamethoxam resistance to determine the extent of the issue in the region., Results: Approximately 57 and 65% of the F. fusca populations surveyed had reduced imidacloprid and thiamethoxam sensitivity respectively. Survivorship in diagnostic bioassays was significantly different at both the state and regional scales. Multiple-dose bioassays conducted on 37 of the populations documented up to 55- and 39-fold resistance ratios for imidacloprid and thiamethoxam respectively., Conclusion: Estimates of neonicotinoid resistance indicate an emerging issue for management of F. fusca in the eastern United States. Significant variation in survivorship within states and regions indicated that finer-scale surveys were needed to determine factors (genetic, insecticide use) driving resistance evolution. © 2016 Society of Chemical Industry., (© 2016 Society of Chemical Industry.)

Plants and their pathogens coevolve locally. Previous investigations of one host-one pathogen systems have demonstrated that natural selection favors pathogen genotypes that are virulent on a broad range of host genotypes. In the present study, we examine a system consisting of one pathogen species that infects three host species in the morning glory genus Ipomoea. We show that many pathogen genotypes can infect two or three of the host species when tested on plants from nonlocal communities. By contrast, pathogen genotypes are highly host-specific, infecting only one host species, when tested on host species from the local community. This pattern indicates that within-community evolution narrows the host breadth of pathogen genotypes. Possible evolutionary mechanisms include direct selection for narrow host breadth due to costs of virulence and evolution of ipomoea resistance in the host species.

Diabetic Charcot neuroarthropathy is a complex, limb-threatening disease process with major lifestyle-altering repercussions for patients. When Charcot neuroarthropathy leads to unstable deformity, ulceration, and potential infection despite conservative therapies, foot and ankle surgeons often consider reconstructive limb salvage procedures to restore function. The purpose of the present study was to evaluate the clinical and radiographic outcomes of diabetic Charcot reconstruction using combined internal and external fixation. A total of 22 patients were reviewed; 16 (72.73%) midfoot and 6 (27.27%) tibiotalocalcaneal arthrodesis procedures were consecutively performed from March 2009 to May 2013. All surgical procedures were performed in nonacute phases of the Charcot process in patients with diagnosed diabetes mellitus and documented peripheral neuropathy. Patients were excluded from the study if they were not diabetic despite having undergone Charcot reconstruction, regardless of the fixation method, or if they did not complete radiographic imaging. During a mean follow-up period of 58.60 ± 42.37 (range 16 to 164) weeks, limb salvage was achieved in 20 patients (90.91%), and 2 (9.09%) required below-the-knee amputation at a mean of 42 ± 14.14 weeks. Wound dehiscence occurred in 8 (36.36%), pin tract infection in 10 (45.45%), and superficial wound infection in 9 (40.91%) and peaked in bimodal fashion at 4 and 8 weeks postoperatively. Radiographic analysis of the pre- versus postoperative alignment showed statistically significant changes in the lateral talo-first metatarsal angle (p = .02) and lateral talar declination angle (p = .01). The limb salvage rates with diabetic Charcot reconstruction are improving in part because of the continued development of increasingly superior modalities for both internal and external fixation., (Copyright © 2016 American College of Foot and Ankle Surgeons. Published by Elsevier Inc. All rights reserved.)

Acylsugars are secondary metabolites exuded from type IV glandular trichomes that provide broad-spectrum insect suppression for Solanum pennellii Correll, a wild relative of cultivated tomato. Acylsugars produced by different S. pennellii accessions vary by sugar moieties (glucose or sucrose) and fatty acid side chains (lengths and branching patterns). Our objective was to determine which acylsugar compositions more effectively suppressed oviposition of the whitefly Bemisia tabaci (Gennadius) (Middle East--Asia Minor 1 Group), tobacco thrips, Frankliniella fusca (Hinds), and western flower thrips, Frankliniella occidentalis (Pergande). We extracted and characterized acylsugars from four S. pennellii accessions with different compositions, as well as from an acylsugar-producing tomato breeding line. We also fractionated the acylsugars of one S. pennellii accession to examine the effects of its components. Effects of acylsugars on oviposition were evaluated by administering a range of doses to oviposition sites of adult whiteflies and thrips in non-choice and choice bioassays, respectively. The acylsugars from S. pennellii accessions and the tomato breeding line demonstrated differential functionality in their ability to alter the distribution of whitefly oviposition and suppress oviposition on acylsugar treated substrates. Tobacco thrips were sensitive to all compositions while western flower thrips and whiteflies were more sensitive to acylsugars from a subset of S. pennellii accessions. It follows that acylsugars could thus mediate plant-enemy interactions in such a way as to affect evolution of host specialization, resistance specificity, and potentially host differentiation or local adaptation. The acylsugars from S. pennellii LA1376 were separated by polarity into two fractions that differed sharply for their sugar moieties and fatty acid side chains. These fractions had different efficacies, with neither having activity approaching that of the original exudate. When these two fractions were recombined, the effect on both whiteflies and thrips exceeded the sum of the two fractions' effects, and was similar to that of the original exudate. These results suggest that increasing diversity of components within a mixture may increase suppression through synergistic interactions. This study demonstrates the potential for composition-specific deployment of acylsugars for herbivore oviposition suppression, either through in planta production by tomato lines, or as biocides applied by a foliar spray.

Background: The codling moth, Cydia pomonella (L.) (Lepidoptera: Tortricidae), is a major worldwide pest of apples, pears and walnuts. A temperature-driven phenological model of codling moth, developed in Michigan, has been utilized in North Carolina and other states for decades. Systematic inaccuracy of this model in predicting moth emergence in North Carolina suggests that the relationship between emergence and temperature differs between the American midwest and southeast, or that additional factors may influence the system., Results: A method was developed to optimize the estimation of generation turnover intervals. Emergence was modeled as a function of heat unit accumulation. Significant differences between emergence predictions based on the resultant model and the existing model developed in Michigan were found., Conclusion: A new model of codling moth emergence, incorporating improved estimates for generation turnover for North Carolina, offers predictive improvement with practical importance to management. Differences between the emergence of susceptible and resistant moth populations were also investigated, leading to the suggestion that resistance to insecticides should be considered in future studies of emergence phenology., (© 2014 Society of Chemical Industry.)

Mutator phenotypes accelerate the evolutionary process of neoplastic transformation. Historically, the measurement of mutation rates has relied on scoring the occurrence of rare mutations in target genes in large populations of cells. Averaging mutation rates over large cell populations assumes that new mutations arise at a constant rate during each cell division. If the mutation rate is not constant, an expanding mutator population may contain subclones with widely divergent rates of evolution. Here, we report mutation rate measurements of individual cell divisions of mutator yeast deficient in DNA polymerase ε proofreading and base-base mismatch repair. Our data are best fit by a model in which cells can assume one of two distinct mutator states, with mutation rates that differ by an order of magnitude. In error-prone cell divisions, mutations occurred on the same chromosome more frequently than expected by chance, often in DNA with similar predicted replication timing, consistent with a spatiotemporal dimension to the hypermutator state. Mapping of mutations onto predicted replicons revealed that mutations were enriched in the first half of the replicon as well as near termination zones. Taken together, our findings show that individual genome replication events exhibit an unexpected volatility that may deepen our understanding of the evolution of mutator-driven malignancies.

Amphibians are suffering unprecedented global declines. A leading cause is the infectious disease chytridiomycosis caused by the chytrid fungus Batrachochytrium dendrobatidis. Chytridiomycosis is a skin disease which disrupts transport of essential ions leading to death. Soluble factors produced by B. dendrobatidis impair amphibian and mammalian lymphocytes in vitro, but previous studies have not shown the effects of these inhibitory factors in vivo. To demonstrate in vivo inhibition of immunity by B. dendrobatidis, a modified delayed-type-hypersensitivity (DTH) protocol was developed to induce innate and adaptive inflammatory swelling in the feet of Xenopus laevis by injection of killed bacteria or phytohemagglutinin (PHA). Compared to previous protocols for PHA injection in amphibians, this method induced up to 20-fold greater inflammatory swelling. Using this new protocol, we measured DTH responses induced by killed bacteria or PHA in the presence of B. dendrobatidis supernatants. Swelling induced by single injection of PHA or killed bacteria was not significantly affected by B. dendrobatidis supernatants. However, swelling caused by a secondary injection of PHA, was significantly reduced by B. dendrobatidis supernatants. As previously described in vitro, factors from B. dendrobatidis appear to inhibit lymphocyte-mediated inflammatory swelling but not swelling caused by an inducer of innate leukocytes. This suggests that B. dendrobatidis is capable of inhibiting lymphocytes in a localized response to prevent adaptive immune responses in the skin. The modified protocol used to induce inflammatory swelling in the present study may be more effective than previous methods to investigate amphibian immune competence, particularly in nonmodel species., (Copyright © 2014, American Society for Microbiology. All Rights Reserved.)

There is no consensus in the literature related to the morphology, bony attachments, and variations of the fibular collateral ligament (FCL) of the knee. Our purpose was to investigate FCL morphology and to review reports in the literature regarding this structure. Seventy knees from formalin-fixed, adult cadavers were dissected and a digital caliper was used to measure FCL length, width, distance from proximal attachment to articular surface (PA→AS), and distance from distal attachment to articular surface (DA→AS). The mean (SE) length and width of all FCLs was 48.3 (1.1) mm and 4 (0.16) mm, respectively. The mean (SE) PA→AS and DA→AS of all FCLs was 22 (0.8) mm and 24.8 (1) mm, respectively. We found a direct relationship between the PA→AS and DA→AS distances (Spearman rho = 0.527, P = 0.002) and this association was independent of age and sex. Two FCL variations were found: a bifurcate ligament with two distal bands and a trifurcate ligament with three distal bands, all of which attached to the fibular head. Our literature review revealed that only 2/10 cadaveric FCL morphology studies reported variations similar to the current study. Further, there was variability in the reported location of the FCL proximal attachment. When combining our data with these studies, 105/219 FCLs directly attached to the apex of the lateral epicondyle (LE), 10/219 to a fovea posterior to the LE, and 104/219 posterior and proximal to the LE. These data may have implications related to FCL injury and repair., (© 2014 Wiley Periodicals, Inc.)

Background: There is heterogeneity in the literature regarding the anatomy and number of ligamentous bands that form the deltoid ligament (DL). Anatomic knowledge of the DL and its variations are important for surgeons who repair ankle fractures., Methods: The DL was dissected in 33 ankles from 17 formalin-fixed cadavers (mean age at death, 76.6 years) to examine its morphology. The length, width, and thickness of its constituent bands were recorded with a digital caliper. Descriptive and correlational statistics were used to investigate the relationships between band size, age at death, and sex. A literature review was conducted to compare our data to those of previous studies., Results: The DL has superficial and deep layers with up to 8 different bands., Conclusion: The DL stabilizes the medial ankle and should be evaluated in flatfoot deformities and severe ankle fractures., Clinical Relevance: Anatomic knowledge of DL variations should aid the surgeon in repairing torn DLs., (© The Author(s) 2014.)

Background: Many cadaver-based anatomy courses and surgical workshops use prosections to help podiatry students and residents learn clinically relevant anatomy. The quality of these prosections is variable and dependent upon the methods used to prepare them. These methods have not been adequately described in the literature, and few studies describe the use of chemicals to prepare prosections of the cadaveric foot and ankle. Recognizing the need for better teaching prosections in podiatric education, we developed a chemical application method with underwater dissection to better preserve anatomic structures of the cadaveric foot and ankle., Methods: We used inexpensive chemicals before, during, and after each step, which ultimately resulted in high-quality prosections that improved identification of anatomic structures relevant to the practice of podiatric medicine., Results: Careful preservation of clinically important nerves, vessels, muscles, ligaments, and joints was achieved with these prosections., Conclusions: Although this method required additional preparation time, the resultant prosections have been repeatedly used for several years to facilitate learning among podiatry students and residents, and they have held up well. This method can be used by educators to teach podiatry students throughout their medical training and even into residency.

Through a modeling approach, we investigated weather factors that affect the summer incidence of Tomato spotted wilt virus (TSWV), a virus vectored exclusively by thrips, in cultivated tobacco. Aspects of thrips and plant biology that affect disease spread were treated as functions of weather, leading to a model of disease incidence informed by thrips and plant biology, and dependent on weather input variables. We found that disease incidence during the summer was influenced by weather affecting thrips activity during the preceding year, especially during a time when thrips transmit TSWV to and from the plant hosts that constitute the virus' natural reservoir. We identified an interaction between spring precipitation and earlier weather affecting thrips, relating this to virus abundance and transmission intensity as interacting factors affecting disease incidence. Throughout, weather is the basic driver of epidemiology in the system, and our findings allowed us to detect associations between atypically high- or low-incidence years and the local climatic deviations from normal weather patterns, brought about by El Niño Southern Oscillation transitions.

We examined the genetic basis of resistance to the rust pathogen Coleosporium ipomoea in three host species: Ipomoea purpurea, I. hederacea, and I. coccinea (Convolvulaceae). In crosses between resistant and susceptible individuals, second-generation selfed offspring segregated in ratios that did not differ statistically from the 3:1 ratio indicative of single-gene resistance with the resistant allele dominant. One out of three crosses between resistant individuals from two different populations revealed that resistance loci differed in the two populations, as evidenced by the production of susceptible individuals among the S(2) generation. These results suggest that gene-for-gene interactions contribute substantially to the dynamics of coevolution in this natural pathosystem. They also suggest that evolution of resistance to the same pathogen strain may involve different loci in different Ipomoea populations.