Honors College Thesis
 

A Genetic Investigation of Proline Induction of Virulence Genes in Pseudomonas syringae DC3000

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https://ir.library.oregonstate.edu/concern/honors_college_theses/6m311x17t

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  • Pseudomonas syringae is a species of bacteria with plant pathogenic members that infect their hosts by colonizing the interior of leaves. Plants can recognize pathogen-associated molecular patterns with receptors and can prevent the growth of pathogens in response. To overcome plant defenses, P. syringae produce defense-suppressing proteins that are delivered into host cells via a type III secretion system (T3SS). P. syringae genes that encode for the T3SS are induced by host chemical signals, yet how these signals are perceived by P. syringae is poorly understood. In this study, we used the transposon Tn5 to randomly mutagenize the genome of P. syringae strain DC3000 and screened 20,000 of the resulting Tn5+ mutants for loss-of-response to proline, a T3SS-inducing amino acid found in the apoplast of Arabidopsis thaliana leaves. The parental strain carried a green fluorescent protein (GFP) transcriptional reporter consisting of the promotor of the T3SS regulator hrpL fused to gfp so we could screen for mutants altered in proline-dependent T3SS expression. From this screen we identified 84 and 22 mutants with reduced and increased expression of GFP, respectively. We identified 76 genes interrupted by the Tn5 transposon, including putA with a predicted role in proline catabolism, as well as several genes with predicted roles in metabolism, signal transduction and transport. Our results suggest that proline catabolism inside the P. syringae cell may be linked to T3SS deployment.
  • Key Words: Type III Secretion System, Pseudomonas syringae, DC3000, transposon mutagenesis, proline, putA
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  • National Science Foundation
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  • Ongoing Research
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  • 2022-06-01 to 2024-07-02

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