Plant Science & Landscape Architecture Theses and Dissertations
Permanent URI for this collectionhttp://hdl.handle.net/1903/2797
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Item SALMONELLA ENTERICA STRATEGIES FOR PERSISTENCE ON TOMATO (SOLANUM LYCOPERSICUM) AND SEROVAR DYNAMICS IN SURFACE AND RECLAIMED WATER(2019) Ferelli, Angela Marie Cecelia; Micallef, Shirley A.; Plant Science and Landscape Architecture (PSLA); Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)While select aspects of Salmonella enterica subspecies enterica persistence in agricultural matrices have been illustrated, serovar specific survival strategies in surface water, transmission, and persistence on plants are multifaceted and remain only partially examined. In the present work, we utilized an interdisciplinary approach to illustrate novel mechanisms by which S. enterica may adapt to plants as an alternative host. Furthermore, we leveraged the wealth of diversity in S. enterica serovars to investigate specific dynamics and drivers of persistence in water and transfer onto produce crops. Through biochemical, gene expression, and plant challenge assays of both tomato (Solanum lycopersicum) vegetative and fruit organs, we found that plant-derived NO was generated in response to S. Newport recognition. Furthermore, bacterial gene expression on both leaves and fruit was indicative of adaptation to a novel environment including upregulation in NO detoxification machinery, indicating plant-derived NO as a novel bacterial stress. NO tolerance of various S. enterica was then evaluated to investigate drivers of “produce associated’ S. enterica adaptation to the plant niche. We identified that plant derived NO can negatively affect titers of all S. enterica serovars tested and that serovar specific tolerance to NO in vitro was apparent in a concentration and exposure time dependent manner. Finally, the survival of various S. enterica in surface and reclaimed water was investigated while evaluating the potential for transition to viable but non-culturable (VBNC) organisms. Furthermore, surface water used for irrigation, a common water environment for S. enterica, was investigated as a priming reservoir for various S. enterica serovars for enhanced transmission onto tomato crops. Persistence in water included VBNC subpopulations and was driven by water type. Transfer success onto tomato was driven by serovar, and prolonged incubation in water increased the transfer ability of serovars that initially transferred poorly onto tomato. Finally, attachment to polystyrene and water oxidation-reduction potential were identified as possible indicators of foodborne pathogen transfer success onto tomato. Moving forward, a greater understanding of the environmental queues used by S. enterica subspecies enterica responding to the agricultural environment will aid researchers in developing S. enterica targeted on-farm management techniques to ensure safe yet sustainable fresh produce cultivation practices.Item Bacterial communities of the specialty crop phyllosphere: response to biological soil amendment use, rainfall, and insect visitation(2016) Allard, Sarah Michelle; Micallef, Shirley A; Plant Science and Landscape Architecture (PSLA); Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)Microorganisms in the plant rhizosphere, the zone under the influence of roots, and phyllosphere, the aboveground plant habitat, exert a strong influence on plant growth, health, and protection. Tomatoes and cucumbers are important players in produce safety, and the microbial life on their surfaces may contribute to their fitness as hosts for foodborne pathogens such as Salmonella enterica and Listeria monocytogenes. External factors such as agricultural inputs and environmental conditions likely also play a major role. However, the relative contributions of the various factors at play concerning the plant surface microbiome remain obscure, although this knowledge could be applied to crop protection from plant and human pathogens. Recent advances in genomic technology have made investigations into the diversity and structure of microbial communities possible in many systems and at multiple scales. Using Illumina sequencing to profile particular regions of the 16S rRNA gene, this study investigates the influences of climate and crop management practices on the field-grown tomato and cucumber microbiome. The first research chapter (Chapter 3) involved application of 4 different soil amendments to a tomato field and profiling of harvest-time phyllosphere and rhizosphere microbial communities. Factors such as water activity, soil texture, and field location influenced microbial community structure more than soil amendment use, indicating that field conditions may exert more influence on the tomato microbiome than certain agricultural inputs. In Chapter 4, the impact of rain on tomato and cucumber-associated microbial community structures was evaluated. Shifts in bacterial community composition and structure were recorded immediately following rain events, an effect which was partially reversed after 4 days and was strongest on cucumber fruit surfaces. Chapter 5 focused on the contribution of insect visitors to the tomato microbiota, finding that insects introduced diverse bacterial taxa to the blossom and green tomato fruit microbiome. This study advances our understanding of the factors that influence the microbiomes of tomato and cucumber. Farms are complex environments, and untangling the interactions between farming practices, the environment, and microbial diversity will help us develop a comprehensive understanding of how microbial life, including foodborne pathogens, may be influenced by agricultural conditions.Item SALMONELLA ENTERICA INTERACTIONS WITH TOMATO: PLANT GENOTYPE EFFECTS AND SALMONELLA GENETIC RESPONSES(2015) Han, Sanghyun; Micallef, Shirley A.; Plant Science and Landscape Architecture (PSLA); Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)Several outbreaks of Salmonella enterica infections have been linked to tomatoes. One cost-effective way to complement on-farm preventive Good Agricultural Practices would be to identify cultivars with inherent decreased susceptibility to Salmonella colonization. Various tomato cultivars with distinct phenotypes were screened to evaluate their susceptibility to Salmonella epiphytic colonization. The potential role of plant exudates, collected from the same cultivars, on the growth kinetics of Salmonella was examined. These investigations were supplemented with Salmonella genome-wide transcriptomics that showed bacterial responses to colonization of tomato shoots and roots. Epiphytic colonization of fruit by S. enterica was cultivar-dependent and serotype-specific, but did not correlate with leaf colonization. Fruit and leaves of the same cultivar differed in their ability to support Salmonella growth. Quantitative and qualitative analysis of tomato exudates provided a possible explanation for the differential susceptibility to bacterial colonization among tomato cultivars. Tomato exudates alone were capable of supporting Salmonella growth, and the growth kinetics of Salmonella in tomato exudates differed by cultivar. Characterization of the chemical composition of primary and secondary metabolites in tomato exudates pointed to potential causes for the differential growth of Salmonella observed in the exudates of various tomato cultivars. Key transcriptomic signals that were down- and up-regulated in Salmonella upon interacting with tomato were identified, enabling us to elucidate the molecular mechanisms underlying this enteric pathogen-plant interaction. Overall, the identified signals lead to a proposed model that depicts the cellular processes needed to preserve cell viability when multiple abiotic stresses in conjunction with low nutrient availability are encountered, while simultaneously repressing unnecessary energy demands or maintaining them at a level equivalent to growth in a nutritious medium. These findings strongly support the hypothesis that plant-regulated mechanisms influence enteric pathogen colonization. It is clear that Salmonella can sense subtle environmental cues brought about by the genotype or physiological state of plants and can respond with distinct patterns of gene expression. Future work should focus on whether this bacterial behavior on plants results from an evolutionary adaptation to use plants as a vector to re-enter animal hosts.