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Shin-Yi  Marzano    Research Molecular Biologist /ARSUserFiles/55604/Marzano, Shin-Yi.jpg **Shin-Yi Lee Marzano, Ph.D.**2801 W. Bancroft St., Mail Stop 604, 4271B Wolfe HallToledo OH 43606Office (419) 530-5053 I have been interested in beneficial microbes that are useful in managing diseases, whether in plant health or human health. I am motivated to study microbiomes and viromes because antimicrobial resistance is a substantial cause of death by infectious disease in the United States, and anti-fungicidal resistance is a big concern in agriculture. With a training in Environmental Microbiology, I am also interested in manipulating microbial pathways that can eliminate nutrient loss in runoff. As part of my postdoc work, I surveyed the fungal virome in *Sclerotinia sclerotiorum*(white mold). The genome of *S. sclerotiorum* has been sequenced and is suitable as a model system from which the most diverse lineages of mycoviruses have been discovered. These mycoviruses include positive- and negative-stranded ssRNA, dsRNA viruses and a ssDNA virus capable of extracellular transmission. My current lab and the others have found that disruption mutants of *S. sclerotiorum* genes are reasonably easy to obtain. Therefore, as a continuation of a previous genome-wide investigation, my lab is dissecting the RNA silencing pathways of *S*. *sclerotiorum*, validating a short list of fungal genes targeted by small RNA-mediated degradation, either induced by mycovirus infection causing hypovirulence (reduced virulence) or by endogenous small RNAs. We approach this line of investigation by establishing reverse genetic systems to look at the effect of virus suppressor of gene silencing (VSR), the effect of different viral infections by synthetic biology, and aiming to optimize the viral systems to deliver transgenes to induce gene silencing (VIGS). Building on what we’ve learned in the past years, we are also interested in learning how secondary small RNAs are produced, and what biological significance they have in fungi. Shin-Yi Marzano Google Scholar The Marzano Lab at USDA-ARS is interested in researching alternative disease control strategies applicable in both controlled environment agriculture and field crops. 1. **Exploring RNAi-based management strategies to confer plant resistance to white mold infection**Growers lack effective genetic tools to manage losses caused by *Sclerotinia sclerotiorum* because of a lack of resistance to the pathogen in germplasms. This necessitates the identification of alternative sources of resistance for the disease. We have identified strong candidate genes in the *Sclerotinia sclerotiorum* RNA silencing pathway as the targets for the development of an RNAi-based pesticide. This study will develop methods for spray-induced gene silencing (SIGS) delivery of small RNAs or dsRNA to reduce the impact of *Sclerotinia sclerotiorum* on some of the dicots. Eventually, we will compare the effectiveness of SIGS on sunflower, edible snapbean, and dry beans for which transgenic approaches such as host-induced gene silencing are not feasible. The **goal** of this project is to develop molecular tools for SIGS control of white mold by three interrelated objectives: **1)** Comparing dsRNA segments generated from different regions of Ago2 and their corresponding RNA secondary structures for SIGS; **2)** Determining the effects of perturbing Ago2 homeostasis by a miRNA with varying base-pairing schemes; **3)** Validating predicted cross-species miRNAs generated from a common mycoparasite that can target genes of members of the family Sclerotiniaceae simultaneously. The proposed work will allow us to better design SIGS for white mold control based on RNA secondary structure and secondary amplification of sRNA. 2. **Developing gemycircularvirus-based pesticide for the control of Sclerotinia sclerotiorum****Sclerotinia sclerotiorum** is a fungal pathogen that causes economically important diseases (e.g., white mold) in several crop species, many of which have not been adequately controlled by conventional technologies. Mycoviruses have been used successfully to reduce losses caused by some fungal plant pathogens. Uniquely, a group of mycovirus, gemycircularviruses, are stable, ubiquitous, and most importantly, infect fungi by extracellular transmission. For example, the prototype, Sclerotinia sclerotiorum hypovirulence-associated DNA virus 1, can control *S. sclerotiorum* as a spray on leaves and has been shown to be field-effective. Unlike many well-studied mycoviruses, there is no need to rely on hyphal-fusion or intracellular transmission. The most well-studied mycovirus, Cryphonectria parasitica hypovirus 1, with an intracellular lifestyle, is not readily transmitted into most of the US strains of *Cryphonectria parasitica*, which has plagued the utility of mycoviruses overall. To overturn the generalized stigma and truly contribute to white mold control, the goal of this study is to determine the fungal host range of Soybean leaf-associated Gemygorvirus-1 (SlaGemV-1) that attenuates fungal virulence, and to develop novel strategies for the delivery of gemycircularvirus-based mycoviral fungicides. We have confirmed the infectivity of SlaGemV-1 on other evolutionarily related fungal pathogens of white mold, such as *Botrytis cinerea* and *Monilinia fruticola*, all belong to Sclerotiniaceae family, but does not infect distantly related fungal pathogens such as *Fusarium oxysporum* and Fusarium *graminearum*. We are currently developing ways to inoculate plants, including various dicots such as tomato, canola, sunflower, dry beans, lettuce, and basil.  **Professional Expereince**TT Assistant Professor (80% research, 20% teaching), 2016-2020. Biology and Microbiology (60%), Agronomy, Horticulture, and Plant Science (40%), South Dakota State UniversityAdjunct faculty, 2020-present, Plant Pathology, Ohio State UniversityAdjunct associate professor, 2020-present, Biological Sciences, University of Toledo **Educational Background**PhD, Crop Sciences, University of Illinois, Urbana-Champaign, USAMSc, Environmental Engineering, University of Illinois, Urbana-Champaign, USAMSc, Environmental Microbiology, Aberdeen University, UKBSc, Environmental Health, National Taiwan University ProjectsSustainable Production and Pest Management Practices for Nursery, Greenhouse, and Protected Culture CropsIn-House Appropriated (D)Accession Number:438340Exploring the Virome and RNAi Strategies for Disease ControlNon-Assistance Cooperative Agreement (S)Accession Number:440636Investigating Effects of Algal Toxins on PlantsNon-Assistance Cooperative Agreement (S)Accession Number:440713Exploring RNAi-Based Management Strategies to Confer Plant Resistance to White Mold InfectionNon-Assistance Cooperative Agreement (S)Accession Number:441603Characterizing Plant- and Virus-Derived Proteins in Important Crops that Enhance the Resistance to White MoldNon-Assistance Cooperative Agreement (S)Accession Number:444401Investigating Leaf Angle Response to Temperature and Carbon DioxideNon-Assistance Cooperative Agreement (S)Accession Number:437048
 Publications(Clicking on the reprint iconReprint Icon  will take you to the publication reprint.)Plants reduced nitrous oxide emissions from northern great plains saline/sodic soil-(Peer Reviewed Journal) Clay, S., Nleya, T., Clay, D., Joshi, D., Bhattarai, D., Marzano, S.L., Petla, B. 2024. Plants reduced nitrous oxide emissions from northern great plains saline/sodic soil. Journal of Environmental Quality. 116(3):1343-1356. https://doi.org/10.1002/agj2.21573.Can phytoremediation-induced changes in the microbiome improve saline/sodic soil and plant health?-(Peer Reviewed Journal) Neupane, A., Jakubowski, D., Fiedler, D., Gu, L., Clay, S., Clay, D., Marzano, S.L. 2023. Can phytoremediation-induced changes in the microbiome improve saline/sodic soil and plant health?. Agronomy Journal. 14(1). Article 29. https://doi.org/10.3390/agronomy14010029.Double-stranded RNA targeting white mold Sclerotinia sclerotiorum argonaute 2 for disease control via spray-induced gene silencing-(Peer Reviewed Journal) Mukherjee, S., Beligala, G., Feng, C., Marzano, S.L. 2024. Double-stranded RNA targeting white mold Sclerotinia sclerotiorum argonaute 2 for disease control via spray-induced gene silencing. Phytopathology. https://doi.org/10.1094/PHYTO-11-23-0431-R.Mechanisms of primed defense: plant immunity induced by endophytic colonization of a mycovirus-induced hypovirulent fungal pathogen-(Peer Reviewed Journal) Pedersen, C.J., Marzano, S.L. 2023. Mechanisms of primed defense: plant immunity induced by endophytic colonization of a mycovirus-induced hypovirulent fungal pathogen. Molecular Plant-Microbe Interactions. 36(11):726-736. https://doi.org/10.1094/mpmi-06-23-0083-r.Characterization of transcriptional responses to genomovirus infection of the white mold fungus, Sclerotinia sclerotiorum-(Peer Reviewed Journal) Pedersen, C.J., Marzano, S.L. 2022. Characterization of transcriptional responses to genomovirus infection of the white mold fungus, Sclerotinia sclerotiorum. Viruses. 14(9). Article #1892. https://doi.org/10.3390/v14091892.Fusarium graminearum virus-1 strain FgV1-SD4 infection eliminates mycotoxin Deoxynivalenol synthesis by F. graminearum in FHB-(Peer Reviewed Journal) Paudel, B., Pedersen, C.J., Yen, Y., Marzano, S.L. 2022. Fusarium graminearum virus-1 strain FgV1-SD4 infection eliminates mycotoxin Deoxynivalenol synthesis by F. graminearum in FHB. Microorganisms. 10: Article 1484. https://doi.org/10.3390/microorganisms10081484.Phytoremediation and high rainfall combine to improve soil and plant health in a North America northern great plains saline sodic soil-(Peer Reviewed Journal) Fiedler, D.J., Clay, S.A., Joshi, D., Westhoff, S., Reese, C., Bruggeman, S., Morelis-Miller, J., Perkins, L.B., Marzano, S.L., Clay, D.E. 2022. Phytoremediation and high rainfall combine to improve soil and plant health in a North America northern great plains saline sodic soil. Journal of Soil and Water Conservation Society. 77(3). Article 00112. https://doi.org/10.2489/jswc.2022.00112.Direct metatranscriptomic survey of sunflower Microbiome and Virome-(Peer Reviewed Journal) Wang, Z., Neupane, A., Feng, J., Pedersen, C., Marzano, S.L. 2021. Direct metatranscriptomic survey of sunflower Microbiome and Virome. Viruses. 13(9). Article 1867. https://doi.org/10.3390/v13091867.Identification of the viral determinant of hypovirulence and host range in Sclerotiniaceae of a genomovirus reconstructed from the plant metagenome-(Peer Reviewed Journal) Feng, C., Feng, J., Wang, Z., Pedersen, C., Wang, X., Saleem, H., Domier, L.L., Marzano, S.L. 2021. Identification of the viral determinant of hypovirulence and host range in Sclerotiniaceae of a genomovirus reconstructed from the plant metagenome. Journal of Virology. 95(17). Article e00264-21. https://doi.org/10.1128/JVI.00264-21.Long term crop rotation effect on subsequent soybean yield explained by soil and root-associated microbiome and soil health indicators-(Peer Reviewed Journal) Neupane, A., Bulbul, I., Wang, Z., Lehman, R.M., Nafziger, E.D., Marzano, S.L. 2021. Long term crop rotation effect on subsequent soybean yield explained by soil and root-associated microbiome and soil health indicators. Scientific Reports. 11. Article 9200. https://doi.org/10.1038/s41598-021-88784-6.CO2 and N2O emissions and microbial community structure from fields that include salt-affected soils-(Peer Reviewed Journal) Fielder, D.J., Clay, D.E., Joshi, D.R., Engel, A., Marzano, S.L., Jakubowski, D., Bhattarai, D., Reese, C.L., Bruggeman, S.A., Clay, S. 2021. CO2 and N2O emissions and microbial community structure from fields that include salt-affected soils. Journal of Environmental Quality. 50(3):567-579. https://doi.org/10.1002/jeq2.20223.Interannual dynamics, diversity and evolution of the virome in Sclerotinia sclerotiorum from a single crop field-(Peer Reviewed Journal) Jia, J., Fu, Y., Jiang, D., Mu, F., Cheng, J., Lin, Y., Li, B., Marzano, S.L., Xie, J. 2021. Interannual dynamics, diversity and evolution of the virome in Sclerotinia sclerotiorum from a single crop field. Virus Evolution. 7(1). Article veab032. https://doi.org/10.1093/ve/veab032.


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