Field-Scale Spatial Distribution and Genotypic Diversity of Sclerotinia Sclerotiorum in Soybeans


Book Description

White mold (WM) in soybeans, caused by Sclerotinia sclerotium (S.s.), is the second most devastating disease of soybeans in the world. The disease causes an average of 2.5% yield loss in soybean production annually in Pennsylvania (PA), which equates to approximately $6,000,000. However, many growers still do not scout for white mold or have not been able to manage it well. My thesis aims to increase our understanding of the biology of S.s. at the field-scale, which will improve sampling strategies, crop loss estimations, and field experimental designs. Studying the genotypic diversity of S.s. populations at a field scale will help to improve management strategies like fungicide applications. The first objective of my thesis was to determine the spatial distribution of WM disease incidence and of S.s. sclerotia in the soil. My second objective was to determine if there was evidence of clonality in field-scale populations of S.s. Third, we determined if the state-scale Pennsylvania population of S.s. was clonal and compared that population to New York and Minas Gerais, Brazilian populations. Fourth, we validated the Sporecaster mobile application for use by soybean growers to forecast WM risk in the Northeast United States. Eight fields were selected for soil sampling of thirty-five quadrats. S.s. sclerotia were manually removed and isolated in the lab. DNA was extracted for 286 isolates and genotyped by fragment analysis. Microsatellite regions of the DNA were amplified at 10 loci and PCR products were analyzed by capillary electrophoresis. The same fields were also scouted to quantify the disease incidence in each quadrat. Lastly, soybean fields in PA and New York were monitored and scouted to conduct a validation of the Sporecaster mobile application for forecasting white mold risk. Across both years, the maximum number of sclerotia in a field was 3.3 sclerotia/kg soil and the maximum disease incidence for a field was 14% due to warmer and drier weather conditions at most locations. The spatial distribution of the pathogen at a field-scale was randomly distributed and only one field showed aggregation. Genotypic results indicated 83 multilocus genotypes were present across PA. Despite a high genotypic diversity, populations at a field scale were clonal and showed little evidence of outcrossing. The Sporecaster mobile application had a two-year average accuracy of 57-74% at predicting white mold disease incidence. Our increased knowledge of the pathogen and the use of the Sporecaster app will help to improve management recommendations and guide future research of white mold management tactics.




Development and Refinement of Integrated Management Strategies for Sclerotinia Stem Rot of Soybean


Book Description

Sclerotinia stem rot (SSR), or white mold, is caused by Sclerotinia sclerotiorum and is one of the top ten yield reducing diseases of soybean worldwide. Complete commercial resistance is confounded by an incomplete understanding of resistance mechanisms and the diversity within S. sclerotiorum populations. Characterization of a diverse isolate collection facilitated the selection of a representative panel of isolates for use in ongoing germplasm resistance evaluations. These isolates were used to further identify candidate germplasm lines exhibiting durable SSR resistance for further integration into commercial soybean cultivars. Monitoring infection of resistant and susceptible lines further helped to understand S. sclerotiorum infection and potential resistance mechanisms. Chemical control is also currently incomplete and, in some cases, unnecessary as disease development requires conditions conducive for simultaneous apothecial germination, ascospore release, and soybean flowering. Multiple site-years of fungicide efficacy trials were evaluated using meta-analyses to identify effective control programs, which will assist growers in making economical management decisions. Risk assessment tools are sometimes used to more accurately predict the timing of effective fungicide applications; however, reliable tools were not available in soybean systems prior to this research. In the SSR pathosystem, dense canopies, cool temperatures, high relative humidity, and moist soil conditions have been shown to favor S. sclerotiorum infection and subsequent disease development. Additionally, studies have shown that apothecial development is sensitive to a narrow range of ultraviolet wavelengths (276-319 nm). Virtual weather data and detailed epidemiological studies have led to the development and validation of models for non-irrigated and irrigated fields which predict apothecial presence based on 30-day averages of mean air temperature, relative humidity, and maximum leaf wetness. These models will be incorporated into a mobile web-based recommendation tool. Moreover, controlled environment and in-field light studies have identified a crucial range of light (295-330 nm) which are required for apothecial development and could be targeted to further improvement of SSR management. Overall, the development of germplasm with broad-spectrum resistance, the study of resistance mechanisms, multiple site-year fungicide efficacy evaluations, and an optimized spray advisory system will provide a new, fresh approach to integrated management of SSR in soybean.




Modernizing Integrated Management Practices for Sclerotinia Stem Rot of Soybean (Glycine Max)


Book Description

Soybean production in the Upper Midwest region of the United States is consistently threatened by Sclerotinia stem rot (SSR), caused by the fungal pathogen Sclerotinia sclerotiorum. Management practices for this disease have historically been studied in depth. However, a reevaluation and modernization of these practices is greatly needed. To achieve this, many methods of control are being examined such as the assessment of integrating multiple cultural practices, the reassessment of chemical controls, the development of improved genetic resistance through both classical breeding and reverse genetics, and the integration of resistant genotypes into predictive models. From this research, the optimum cultural and chemical practices were identified for controlling SSR across the Midwest. These practices were examined for their effect on partial profits to understand their economic feasibility. Through breeding efforts, three elite soybean genotypes were identified with high levels of resistance to SSR, and more early-generation breeding lines are being progressed from additional breeding populations. Furthermore, four breeding lines were identified as having differential consistent responses to S. sclerotiorum, and these four lines were established as standardized checks. Using these four check lines, the integration of resistance into S. sclerotiorum apothecial models is being studied to improve predictive accuracy. Collectively, these research efforts will help to modernize management and control of SSR from both research and production perspectives.







Development of Multiple Resistance Strategies to [Sclerotinia Sclerotiorum] in Soybean ([Glycine Max])


Book Description

Soybean resistance to the necrotrophic pathogen [Sclerotinia sclerotiorum] is incomplete, and yield losses from Sclerotinia stem rot (SSR) can surmount to over 300 kg/ha for every 10% increase in SSR severity. Through this work, I aimed to enhance resistance to SSR through novel breeding methods and RNAi. Germplasm identified through our selections in the greenhouse and disease nurseries have served as valuable sources of high physiological resistance to SSR and resulted in a food grade soybean. Subsequent crosses integrating this germplasm aimed to enhance the agronomics of resistant parents and have been expediated through winter selections in the greenhouse and Chile. Additionally, the pathogenic success of [S. sclerotiorum] requires the secretion key virulence factors such as oxalic acid (OA). Virus-induced gene silencing (VIGS) using Bean pod mottle virus (BPMV) was used to target OA biogenesis in [S. sclerotiorum]. A sequence of 366 bp, corresponding to the fungal [oxaloacetate acetylhydrolase] ([Ssoah1]), was cloned into a BPMV vector and biolistically introduced into [Glycine max]. Plants containing BPMV vectors targeting [Ssoah1] substantially enhanced resistance to [S. sclerotiorum] and lowered expression of [Ssoah1] compared to empty-vector control plants in three replicated experiments (P




Trichoderma as a Biocontrol Agent Against Sclerotinia Stem Rot Or White Mold on Soybeans in Brazil


Book Description

Biological control agents are alternatives to chemical pesticides in the management of plant diseases. Currently, hundreds of bioproducts are commercially available in international market varying mainly in antagonistic microorganisms and formulation. We screened four Trichoderma-based products as to their efficacy in controlling Sclerotinia stem rot (SSR) under protected and field environments and their effect on soybean seeds,Äô sanity and physiological qualities. We also tested application technologies through seed microbiolization and foliar spraying to deliver the microorganisms, and their compatibility with chemical fungicides. In vitro assays showed that all Trichoderma strains were antagonistic to S. sclerotiorum evidencing hyperparasitic activity. Moreover, the bioproducts reduced fungi incidence on soybean seeds, promoted faster seedling emergence and did not hamper seeds,Äô vigor. Increases of 14 and 37% were registered for root length and shoot fresh weight over that of the untreated control indicating potential application of the bioproducts as soybean growth promoters. Thiophanate-methyl and procymidone were the most compatible, without drastically affecting spore germination or mycelium growth. Under field conditions, all Trichoderma strains reduced SSR incidence and increased soybean grain yield. Formulation interferes on bioproducts,Äô viability and efficacy deserving special attention upon development.




Seed Transmission and Control of Sclerotinia Sclerotiorum in Soybean Seeds


Book Description

The fourth objective was the seed conditioning as a part of disease management. The fungus was analyzed in relation to the seed size. Seeds that pass through a 10/64 sieve represent the fraction of seeds with the highest incidence of infection. The seed conditioning process (particularly the air screen cleaner) can be very effective in eliminating sclerotia, which represents an important source of inoculum for Sclerotinia sclerotiorum, The fifth objective was to investigate the effectiveness of seed treatment fungicides in eradicating seedborne infection of soybeans with Sclerotinia sclerotiorum. The best results were obtained using carboxim + thiram which reduced the fungus expression by 99% for inoculated seeds, and captan + PCNB + thiabendazole, which reduced the fungus expression by 89% for inoculated seeds.