Frogeye leaf spot, caused by Cercospora sojina Hara, is a foliar disease affecting soybean (Glycine max (L.) Merr.), often managed by applications of quinone outside inhibitor (QoI) fungicides. In 2013 and 2014, symptomatic leaf samples were collected from Mississippi soybean fields leading to the collection of 634 mono-conidial C. sojina isolates. In vitro bioassays were performed to evaluate the sensitivity of 14 isolates plus a baseline. Resistant and sensitive isolates were characterized by determining the effective fungicide concentrations at which 50% of conidial germination was inhibited (EC50). Additionally, the molecular mechanism of resistance was determined for all 634 isolates. Greater than 93% of C. sojina isolates collected in Mississippi carried the G143A amino acid substitution indicating a shift to a QoI-resistant population throughout Mississippi soybean fields. Greenhouse studies confirmed that due to this amino acid substitution, symptoms caused by QoI-resistant isolates developed in spite of a QoI fungicide application.

Soybean is an economically important crop. It is a selfertilized species grown on vast contiguous acres. These facts predispose soybean to disease epidemics. Cercospora sojina, causal agent of frogeye leaf spot, has reduced United States soybean productivity 0.3 percent on average per year between 2008 and 2010. Several states have reported the pathogen developing resistance to the strobilurin class of fungicides. To date genetic host resistance has been identified as single dominant genes (Rcs1, Rcs2, and Rcs3). However, the lifespans of Rcs1 and Rcs2 were 10 and 16 years respectively. Currently, the Rcs3 locus has been utilized in all major soybean breeding programs of the US and has been for over 20 years. Seventyive accessions of soybean were found to exhibit resistance to multiple races of C. sojina while not exhibiting the Rcs3 haplotype. Twenty of these plant introductions (PIs) were screened by six races within the new race classification system of C. sojina representing all domestic variability of the pathogen. Two agronomically favorable PIs, PI398993 and PI399068, were found in this research to exhibit broad resistance to sources documented to contain most domestic variability of the pathogen. Two segregating populations were developed by crossing PI398993 x ‘Blackhawk’ and PI399068 x Blackhawk. Segregation ratios of F2 as well as F2:3 family seedling screens of both populations indicating single dominant gene action in both resistance sources. Single marker analysis indicated markers associated with the phenotype were indeed on chromosome 16 (MLG J), but possibly beyond Rcs3 in both sources. Interval mapping placed the highest probability of the resistance loci near SNP_171 and SNP_368, 72.86 and 72.48 cM respectively, but distal to the Rcs3 locus. Analysis of reaction ratings also indicated significant influence on phenotype was also associated with markers located at or beyond the published Rcs3 locus. The evidence in this research supports the hypothesis that both PIs may contain a resistance loci, potentially different than Davis, but within the same gene cluster. Equally as likely, the resistance could prove allelic to Davis.

Cercospora sojina, the causal agent of frogeye leaf spot, is an important pathogen of soybean that reduces soybean yield by an average of 9.1 million bushels each year in the United States. Management strategies include using resistant cultivars and fungicide applications. Hill plots were used to evaluate over 600 commercial and public varieties for resistance or susceptibility to C. sojina. Each hill plot was spaced 0.6 m apart, planted with 10 seeds from a single cultivar, and was replicated three times. At flowering, the plants were inoculated by spraying a spore solution across all plants in the field. Hill plots were rated for disease severity at 14, 28 and 35 days after inoculation. There were 259 varieties that were susceptible to FLS, with 181 varieties with intermediate resistance, and 161 resistant varieties. A greenhouse trial evaluated fungicides and their ability to hinder symptom production and the duration of this suppression. Spores of the pathogen were applied at 7, 14 and 21 days after application of fungicides. Disease ratings were collected at 5 weekly intervals following inoculation. The significantly different treatment with the lowest disease ratings was the fungicide Stratego YLD (a strobilurin and triazole mix). The fungicide Tilt was found to be significantly different from all other treatments in the number of lesions on a single tracked leaf, including the non-treated control. These results can be used to recommend to producers available varieties.

Frogeye leaf spot is a foliar disease of soybean caused by Cercospora sojina Hara, which until recently had been successfully managed by quinone outside inhibitor (QoI) fungicides. After widespread resistance to the QoI fungicides was reported throughout Mississippi, the next step in characterizing C. sojina was to study the fitness of selected isolates. Fitness measurements of resistance stability, colony growth, conidia production and germination, and virulence were assessed. A phylogenetic analysis was also conducted to assess the genetic similarity of the QoI-resistant and -sensitive C. sojina isolates. All isolates remained stable in terms of QoI resistance. Results of all fitness measurements indicated no significant differences between the QoI-resistant and -sensitive C. sojina isolates. The phylogenetic analysis supported these results revealing similarity between QoI-resistant and -sensitive C. sojina isolates. Based on these results no indication of a fitness cost is associated with QoI resistance in C. sojina isolates from Mississippi soybean.

Fungi in the genus Trichoderma have been characterized as biocontrol agents of plant pathogens since the 1930s. The use of biologicals for disease management has increased in recent years, typically marketed as a safer alternative to chemical applications. However, biologicals often lack consistent control across varying environmental conditions. To overcome the loss in efficacy due to environmental conditions, biologicals can be combined with common fungicide seed-treatments to provide improved control. Additionally, the presence of a biological organism could slow the development of a pathogen population. Greenhouse trials were conducted to determine the baseline root colonization of three Trichoderma spp. used in conjunction with five commonly used seed treatments. In field trials, a stand-alone treatment of the Trichoderma isolates was assessed for management of Rhizoctonia root rot (caused by Rhizoctonia solani) and frogeye leaf spot (caused by Cercospora sojina). The greenhouse trial provided evidence that isolates of T. virens and T. hamatum can colonize the roots of plants in which seeds were treated with metalaxyl + prothioconazole + penflufen or metalaxyl + prothioconazole + penflufen + fluopyram. Surprisingly, in the Rhizoctonia root rot trials, the soybean seedlings treated with Trichoderma spp. had significantly reduced stand compared to the R. solani inoculated control. For the frogeye leaf spot trial, an application of T. virens conidial suspensions as a foliar treatment significantly (P ≤ 0.10) reduced frogeye leaf spot severity of soybean compared to a non-treated control. Future research is warranted to better understand the potential efficacy in additional environments and the mechanism(s) of action used by the Trichoderma isolates evaluated in these experiments.

Soybeans (Glycine max L.) are an important crop globally for its food, feed, and oilpurposes. It is impacted by many diseases, including Cercospora sojina, the causal agent of Frogeye Leaf Spot (FLS). Chemical and cultural controls to this fungal pathogen are insufficient, so genetic resistance must be acquired for adequate control. To this end, two recombinant inbred populations were screened in a greenhouse setting for their relative resistance to FLS, and their genomes were analyzed for contributing quantitative trait loci (QTL). In the Essex ́ Forrest population, one QTL was discovered on chromosome 13, and in the Forrest ́ Williams 82 population, two QTL were identified on chromosomes 6 and 11, respectively. These populations were then also screened in a field setting for agronomic traits. These traits were analyzed to detect one superior line for both FLS resistance and advanced agronomic traits, F ́W 125. This line should be used in future breeding projects to increase FLS resistance and reduce linkage drag for other desired characteristics.

Soybean is an important source of protein in animal feed, and growing demand for meat consumption worldwide has led to increased soybean production. Over 120 million metric tons of soybean were harvested in the United States in 2018, approximately one-third of the world production. In the Mid-Atlantic region, soybean is one of the most valuable field crops. Major foliar diseases that reduce soybean yield in the Mid-Atlantic region are frogeye leaf spot (FLS) and Cercospora leaf blight. In addition to crop rotation and host resistance, foliar fungicides, often with quinone outside inhibitor (QoI) active ingredients, are used to manage these soybean foliar diseases. Yield benefits of foliar fungicides have been inconsistent and this may be the result of low disease pressure, unfavorable environmental conditions for disease development, or the presence of fungal pathogen populations that have developed resistance to fungicides. The objectives of this research were 1) to develop a pyrosequencing-based assay to rapidly quantify QoI resistance frequencies in Cercospora sojina, the causal agent of FLS, 2) to examine the effects of fungicide application timings, disease pressure, and environmental factors on soybean yield, and 3) to develop a weather-based soybean foliar fungicide application decision aid for the Mid-Atlantic U.S. using a threshold decision rule. A pyrosequencing assay targeting the G143A mutation was designed, and a Virginia survey of C. sojina populations indicated that the G143A mutation conferring QoI resistance is widespread. In small plot fungicide application timing experiments, five weekly fungicide applications starting at beginning pod (R3) resulted in the greatest yield, but for single fungicide applications, R3 or 1 week after R3 resulted in the greatest yields. There was positive relationship between the cumulative number of disease favorable days (mean daily temperature 20-30°C and ≥ 10 hours of relative humidity >90%) from planting to R3 and disease severity at the full pod stage (r = 0.97, P = <0.01). Higher disease severity was associated with greater yield loss (r2 =0.53, P = 0.10) suggesting foliar fungicide applications are more likely to have yield benefits as the number of disease favorable days prior to R3 increase. A disease favorable-days threshold (FDT) using the environmental parameters indicated above was evaluated in on-farm experiments throughout Virginia, Maryland, and Delaware. Based on decision rules, FDT = 8 three weeks prior to R3 was the best predictor of a yield benefit with an R3 fungicide application. The decision aid was also able to correctly predict when a fungicide application would not be profitable ≥90% of the time. This weather-based decision aid along with monitoring of fungicide resistance development within the region will provide soybean growers in the Mid-Atlantic U.S. with tools to maximize yields and profitability. / Doctor of Philosophy / Soybean is the third most valuable field crop in the world, ranked only behind rice and wheat in value. Over 98% of the soybean crop is used for animal feed due to its high protein content. The United States is the largest soybean producer in the world, responsible for one-third of global production. Soybean is the top cash crop in the Mid-Atlantic region. Foliar fungal diseases can reduce the soybean yield by causing lesions on the leaves that reduce photosynthesis and cause premature defoliation. Frogeye leaf spot (FLS) caused by Cercospora sojina is a major yield reducing soybean foliar diseases in the Mid-Atlantic region. Foliar fungicides, often with quinone outside inhibitor (QoI) active ingredients, are used to manage the disease. However, fungicide efficacy has been inconsistent. Inconsistencies may be due to low disease pressure, improper application timing, or fungicide resistance. The purpose of this research was to investigate the fungicide efficacy inconsistencies and to develop management tools to improve yield and maximize profitability. Our objectives were to 1) develop a molecular assay to quantify frequencies of the mutation conferring fungicide resistance in Virginia populations of C. sojina, 2) examine the effects of fungicide application timings, disease severity, and weather on soybean yield, and 3) develop a weather-based soybean foliar fungicide application decision aid for the Mid-Atlantic U.S. The C. sojina fungicide resistance mutation was widespread in Virginia, but overall frequencies were relatively low compared to findings from Midwest and Southern states. In fungicide timing experiments, beginning pod (R3) applications resulted in the most consistent yield benefits, and disease severity and yield loss increased as the number of weather-based disease favorable days prior to R3 increased. We used data from on-farm experiments in Virginia, Maryland, and Delaware to develop a weather-based disease favorable-days threshold that increased the probability that a fungicide application at R3 would have a yield benefit in soybean. The results of our research have led improved fungal disease management recommendations for soybean in the Mid-Atlantic that will maximize yields and profitability.

La combinación de la siembra directa con el monocultivo, junto con los cambios climáticos y la difusión de genotipos susceptibles, han generado las condiciones óptimas para que los patógenos necrotróficos, que sobreviven en los rastrojos sean anualmente responsables por daños y pérdidas significativas en el cultivo de soja. Dentro de este grupo de enfermedades, deben destacarse por su prevalencia y severidad, a las enfermedades de fin de ciclo (EFC) y a la mancha ojo de rana (MOR), Entre las prácticas para su manejo, deben mencionarse a la rotación de cultivos, tratamiento químico de semillas, resistencia genética, y el uso de fungicidas en órganos aéreos. Esta última medida se ha incrementado considerablemente como respuesta al aumento del potencial de inóculo sobre variedades susceptibles y bajo monocultivo. Sin embargo, la determinación del momento oportuno de la aplicación de fungicidas para el manejo de estas enfermedades no resulta una tarea sencilla y práctica. Existe una gran complejidad en el abordaje de su estudio con variadas dificultades que deben enfrentarse principalmente cuando el objetivo es asistir a los productores y asesores con metodologías prácticas que aseguren la sustentabilidad económica y ambiental. A pesar de que muchos investigadores están de acuerdo en que los fungicidas deberían ser aplicados durante el período crítico de generación del rendimiento del cultivo de soja, no fue posible encontrar información científica publicada sobre el momento más apropiado de aplicación de fungicidas, dentro de esa amplia ventana fisiológica de aplicación, que relacione ambiente, epidemiología, daños y umbrales. Los objetivos de este trabajo fueron a) generar criterios de uso de fungicidas para el manejo químico sustentable de las EFC y la MOR en el cultivo de soja, b) cuantificar sus daños, c) elaborar y validar un sistema de puntuación que permita definir el momento más eficiente de aplicación para las EFC, y d) determinar los umbrales de control para la MOR. Para ello, se realizaron y analizaron numerosos ensayos de campo provenientes de zonas agro-ecológicas diferentes. Para el caso de las EFC, los resultados demostraron que la respuesta al uso de fungicidas (kg/ha) dependió de la ocurrencia de lluvias entre R3-R5 y no de las que ocurrieron entre R1-R3, por lo tanto dependiendo de las condiciones ambientales, la aplicación de fungicida podría realizarse en R3, R4 o R5. Una vez que se logró identificar el factor ambiental y el período fenológico que estadísticamente se asociaron con el impacto en los rendimientos por el uso de fungicidas, la siguiente etapa fue la de desarrollar un sistema de puntuación basado en la ponderación de diferentes factores agronómicos. Dicho sistema contó con 10 factores de valoración relativa y permitió proponer una herramienta fácil, práctica y dinámica para orientar la decisión de control químico. Los ensayos de validación del sistema de puntuación llevados a cabo en localidades y campañas agrícolas diferentes, permitieron analizar el impacto en el rendimiento agronómico y en la rentabilidad lograda, cuando se utilizó el sistema de puntuación en comparación con las aplicaciones fenológicas fijas de R3, R5 y con un testigo sin aplicación química. Cuando el sistema recomendó no aplicar, los rendimientos que fueron obtenidos en los tratamientos de R3 y R5 no se diferenciaron de los del testigo, indicando que no se justificaba aplicar. Por otro lado, se demostró que algunos ensayos mostraron el mayor impacto por el uso de fungicidas cuando se realizaron las aplicaciones en R3 ó R4 ó R5 y que su decisión final dependió de la puntuación arrojada por el sistema en cada ensayo, demostrando que la aplicación regida exclusivamente por fenología rígida y estricta, no resulta en una alternativa técnica fundamentada para orientar el momento de la pulverización. Para el estudio de la MOR, se lograron estimar los daños y generar recomendaciones para su manejo mediante umbrales de control. El número de lesiones por folíolo central constituyó una medida objetiva, rápida y sencilla para cuantificar la epidemia y desarrollar los diferentes UDE. De esta manera los resultados epidemiológicos obtenidos indican que la MOR no debe ser considerada una clásica EFC debido a su comportamiento policíclico. Para diferentes estadios reproductivos de la soja y para cada estrato foliar considerado, se propusieron umbrales de daño móviles que serán de utilidad para enmarcar el uso de fungicidas dentro de un programa de manejo Integrado.

No description available.

<p>  </p> <p>Protecting crops from disease requires continuous research because plant pathogen incidence, geographical range, and pathogenicity, are constantly shifting variables as agronomic practices and climate continue to evolve. The objectives of this research are to i.) evaluate field-scale fungicide timing programs for corn (<em>Zea mays L.</em>) diseases at multiple locations in Indiana; ii.) evaluate field-scale fungicide timing programs for soybean (<em>Glycine Max</em> (L.) Merr.) diseases at multiple locations in Indiana; iii.) continue to identify, document, and confirm the distribution of populations of the soybean frogeye leaf spot pathogen (<em>Cercospora sojina)</em> that contain the G143A mutation conferring resistance to quinone outside inhibitor (QoI) fungicides in Indiana; and iv.) assess the incidence, severity, and prevalence of tar spot (<em>Phyllachora maydis</em>) in Indiana. For the first and second objectives, field scale trials were established at three locations in Indiana from 2019 to 2022. No application timings at any location provided significant yield protection for corn or soybeans. To achieve the third objective, 165 isolates of <em>C. Sojina </em>were tested. In total, 24 out of the 32 counties sampled in 2021 and 2022 were documented with QoI-resistance. The fourth objective was accomplished by surveying Indiana counties for incidence and severity of tar spot. As of 2022, 86 out of 92 Indiana counties have been confirmed for the presence of tar spot.</p>