College of Agriculture & Natural Resources
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The collections in this community comprise faculty research works, as well as graduate theses and dissertations.
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Item SPRAY STRATEGIES AND SELECTION FOR FUNGICIDE RESISTANCE: FENHEXAMID RESISTANCE IN BOTRYTIS CINEREA AS A CASE STUDY(2023) Boushell, Stephen Carl; Hu, Mengjun; Plant Science and Landscape Architecture (PSLA); Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)Fungicide resistance is a limiting factor in sustainable crop production. Despite the wide adoption of general resistance management strategies by growers, the recent rate of resistance development in important fungal pathogens is concerning. In this study, Botrytis cinerea and the high-risk fungicide fenhexamid were used to determine the effects of fungicide dose, tank mixture, and application timing on resistance selection across varied frequencies of resistance via both detached fruit assays and greenhouse trials. The results showed that application of doses lower than the fungicide label dose, mixture with the low-risk fungicide captan, and application post-infection seem to be the most effective management strategies in our experimental settings. In addition, even a small resistant B. cinerea population can lead to a dramatic reduction of disease control efficacy. Our findings were largely consistent with the recent modeling studies which favored the use of the lowest possible fungicide dose for improved resistance management.Item STRUCTURE-FUNCTION ANALYSES OF AN ESSENTIAL VIRULENCE DETERMINANT OF THE LYME DISEASE PATHOGEN(2022) Foor, Shelby Dimity; Pal, Utpal; Veterinary Medical Science; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)Lyme Disease (LD) is a tick-borne disease caused by a group of gram-negative-like spirochetal pathogens called Borrelia burgdorferi sensu lato. The number of cases reported in the United States have dramatically increased with CDC estimating 476,000 cases annually. This multifaceted infection can spread throughout the entire body, causing clinical complications of the central nervous system, joint and heart. Early antibiotic treatment is available and effective; however, untreated patients can develop chronic symptoms, and even after antibiotics, symptoms of unknown etiology and pathogenesis can develop into post-treatment Lyme disease syndrome (PTLDS). The enzootic life cycle of B. burgdorferi is maintained typically between a small rodent and the Ixodes tick vector, where transmission occurs during tick feeding on a host. Infection establishes after B. burgdorferi is deposited in the dermis and undergoes the required shift in its protein expression profile necessary to support spirochete persistence and pathology, often highlighting protein targets for development of diagnostic, therapeutic, and preventative measures. Two such proteins identified, BB0238 and BB0323, serve as novel virulence determinants and are essential for mammalian infection. These two proteins directly interact, mutually stabilize each other post-translationally, and form an essential complex required for infection; however, their precise functions remain undetermined. In collaborative efforts, we predicted a two-domain structure of BB0238. The N-terminal domain was predicted by AI methods to harbor an antiparallel helix-turn-helix motif (HTH) followed by a third helix and a low-confidence predicted meandering segment. The C-terminal domain structure was determined by X-ray crystallography as well as predicted with high confidence to adopt an α+β fold that resembles closely that of the nuclear transport factor 2 (NTF2) superfamily. While full-length BB0238 lacks homology to singular proteins of known functions, the individual N- and C-terminal regions display structural homology to non-bacterial proteins, particularly to eukaryotic sorting, or transport proteins, suggesting that BB0238 supports an unconventional function in spirochetes. We discovered that BB0238 binds another borrelial protein annotated as BB0108, orthologs of two bacterial chaperones and foldases, the extracellular membrane anchored PrsA, and the periplasmic SurA. This identified interaction requires further investigation, however, may be important for BB0238 protein stability or assist with the novel BB0238 function discovered herein, which regulates proteolytic processing of BB0323. Furthermore, We show that key amino acid residues within the HTH stabilize BB0238 in an environment-specific manner, influence its oligomerization properties, and facilitate tick-to-mouse transmission by aiding spirochete evasion of host cellular immunity, underscoring BB0238’s ability to support microbial establishment during early mammalian infection. Together, these studies highlight the divergent evolution of multidomain spirochete proteins involved in multiplex protein-protein interactions, possibly facilitating multiple functions, which support pathogen survival and thus, represent novel targets for vaccine and therapeutic development against Lyme disease.Item The immunoregulation of interleukin-27 in African trypanosome infection(2018) Liu, Gongguan; Shi, Meiqing; Veterinary Medical Science; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)Interleukin (IL)-27 is a cytokine with diverse impacts on regulation of vertebrate T helper Type 1 (Th1) responses. Initially, it was predicted as a promoter of Th1 responses. However, it was lately identified as a potent negative regulator of T cell responses in a variety of disease models, including infection with viruses, bacteria, and intracellular parasites. The extracellular protozoan parasites, African trypanosomes, cause a chronic debilitating disease associated with persistent inflammation. Using this infection model, we aim to identify novel immunoregulatory functions of IL-27 on innate and adaptive immunity. Here we demonstrate that IL-27 receptor deficient (IL-27R-/-) mice infected with African trypanosomes display excessive production of IFN-γ by CD4+ T cells, exacerbated liver pathology, and dramatically shortened survival as compared with infected wild-type mice. Depletion of CD4+ T cells or neutralization of IFN-γ ameliorates the liver pathology and extends the survival of infected IL-27R-/- mice. Our further interest is in deciphering the mechanisms of how CD4+ T cells and IFN-γ shape the monocyte-featured innate immunity in African trypanosome infected IL-27R-/- mice. Blood monocytes typically consist of a heterogenous population of Ly6C+ and Ly6C- monocytes. Ly6C+ monocytes can give rise to inflammatory TNF-α/iNOS producing dendritic cells (Tip-DCs) and anti-inflammatory macrophages. Here we find that IL-27R-/- mice exhibit a higher frequency of Ly6C+ monocytes recruitment to the liver, where they preferentially differentiate into Tip-DCs. This is coincided with impaired development of Ly6C- monocytes and macrophages in the liver. Depletion of CD4+ T cells or neutralization of IFN-γ in infected IL-27R-/- mice diminishes the recruitment of Ly6C+ monocytes, and their differentiation into Tip-DCs in the liver. This is accompanied by the greatly enhanced counts of Ly6C- monocytes and macrophages following antibody treatments. Further evidences show that 1) IFN-γ produced by CD4+ T cells induces cell death of Ly6C- monocytes which perturb the development of Tip-DCs in infected IL-27R-/- mice and 2) cell intrinsic IFN-γ signaling drives Ly6C+ monocytes to differentiate into Tip-DCs in infected IL-27R-/- mice. Thus, our data identify IL-27 signaling as a novel immunoregulator to prevent Ly6C+ monocytes from differentiation into Tip-DCs through suppressing CD4+ T cells to secrete IFN-γ.Item EXPLORING IL-7R-ALPHA DEVELOPMENTAL EFFECTS AND ONCOGENIC COLLABORATIONS(2017) Cramer, Sarah Delia; Samal, Siba; Veterinary Medical Science; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)Acute lymphoblastic leukemia is the most common cancer of children. Individual cases of leukemia may have multiple genetic lesions, and identifying those that drive leukemogenesis will be important in the development of targeted therapy. Approximately 10% of pediatric T-cell acute lymphoblastic leukemia (T-ALL) cases have a mutation in IL-7Rα. These mutations are thought to be oncogenic, but little is known about the effects of the mutation on T-cell development. In addition, the mutation does not seem to induce leukemia in the absence of other genetic lesions, suggesting that collaborative mutations are required for leukemogenesis. Based on patient data, potential collaborators include TLX3 expression, HOXA gene cluster overexpression, and NRAS mutation. Given the current state of knowledge regarding mutant IL-7Rα, this project was developed with two specific aims. The first was to investigate the effects of mutant IL-7Rα gain-of-function (IL-7Rα-GOF) on T-cell development in vitro and in vivo. The second was to determine whether candidate collaborative genetic lesions would drive T-ALL formation when combined with mutated IL-7Rα. To address these aims, immature murine thymocytes were cultured on an OP9-DL4 stromal cell system, transduced with retroviral vectors, and injected into sub-lethally irradiated Rag1-/- mice. Resultant diseases were analyzed using a variety of techniques including flow cytometry, histology, immunohistochemistry, ligation-mediated PCR, TCRβ clonality assessment, RNA-sequencing, serial passage, and limiting dilution assay. Studies showed that IL-7Rα-GOF mutation caused an increase of CD8+ cells in vitro. When thymocytes transduced with IL-7Rα-GOF mutation were injected into mice, animals developed a multi-systemic inflammatory disease. This inflammation was not due to imbalance in populations of Treg and Th17 cells, as had been hypothesized. Assessing collaborations with TLX3 expression, HOXA overexpression, and NRAS mutation showed that combination of these genetic lesions with IL-7Rα-GOF mutation caused different neoplastic diseases. The combination of IL-7Rα-GOF mutation and TLX3 expression caused low-penetrance, late-onset T-cell lymphoma. Thymocytes overexpressing the HOXA gene cluster and transduced with IL-7Rα-GOF mutation caused a rapid-onset myeloid leukemia. Combination of IL-7Rα-GOF mutation with mutant NRAS yielded rapid-onset, full-penetrance T-cell lymphoblastic leukemia, suggesting that this combination of mutations was sufficient to induce T-ALL. These experimental results may help to lay the foundation for the development of targeted therapy for pediatric T-ALL.Item UNDERSTANDING LATE SEASON FRUIT ROT PATHOSYSTEMS AND INSECT INTERACTIONS IN MID-ATLANTIC VINEYARDS(2016) Kepner, Cody; Swett, Cassandra L; Plant Science and Landscape Architecture (PSLA); Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)Fungal fruit rots and insect pests are among the most important problems negatively affecting the yield and quality of mid-Atlantic wine. In pathogenicity trials of fungi recovered from diseased Chardonnay and Vidal blanc grapes, Alternaria alternata, Pestalotiopsis telopeae, and Aspergillus japonicus were found to be unreported fruit rot pathogens in the region. Additionally, P. telopeae and A. japonicus had comparable virulence to the region’s common fruit rot pathogens. Furthermore, a timed-exclusion field study was implemented to evaluate vineyard insect-fruit rot relationships. It was found that clusters exposed to early-season insect communities that included Paralobesia viteana had a significantly greater incidence of sour rot than clusters protected from insects all season. These results were contrary to the current assumption that fall insects are the primary drivers of sour rot in the region. This research provides diagnostic tools and information to develop management-strategies against fungal and insect pests for mid-Atlantic grape growers.