Browsing by Author "Duong, Duy An"

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    Analyzing the Transcriptomes of Two Quorum-Sensing Controlled Transcription Factors, RcsA and LrhA, Important for Pantoea stewartii Virulence
    Burke, Alison K.; Duong, Duy An; Jensen, Roderick V.; Stevens, Ann M. (Public Library of Science, 2015-12-23)
    The Gram-negative proteobacterium Pantoea stewartii subsp. stewartii causes wilt disease in corn plants. Wilting is primarily due to bacterial exopolysaccharide (EPS) production that blocks water transport in the xylem during the late stages of infection. EsaR, the master quorum-sensing (QS) regulator in P. stewartii, modulates EPS levels. At low cell densities EsaR represses or activates expression of a number of genes in the absence of its acyl homoserine lactone (AHL) ligand. At high cell densities, binding of AHL inactivates EsaR leading to derepression or deactivation of its direct targets. Two of these direct targets are the key transcription regulators RcsA and LrhA, which in turn control EPS production and surface motility/adhesion, respectively. In this study, RNA-Seq was used to further examine the physiological impact of deleting the genes encoding these two second-tier regulators. Quantitative reverse transcription PCR (qRT-PCR) was used to validate the regulation observed in the RNA-Seq data. A GFP transcriptional fusion reporter confirmed the existence of a regulatory feedback loop in the system between LrhA and RcsA. Plant virulence assays carried out with rcsA and lrhA deletion and complementation strains demonstrated that both transcription factors play roles during establishment of wilt disease in corn. These efforts further define the hierarchy of the QS-regulated network controlling plant virulence in P. stewartii.
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    Complete Genome Assembly of Pantoea stewartii subsp. stewartii DC283, a Corn Pathogen
    Duong, Duy An; Stevens, Ann M.; Jensen, Roderick V. (2017-06)
    The phytopathogen Pantoea stewartii subsp. stewartii DC283 causes Stewart's wilt disease in corn after transmission from the corn flea beetle insect vector. Here, we report that the complete annotated genome of P. stewartii DC283 has been fully assembled into one circular chromosome, 10 circular plasmids, and one linear phage.
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    Integrated downstream regulation by the quorum-sensing controlled transcription factors LrhA and RcsA impacts phenotypic outputs associated with virulence in the phytopathogen Pantoea stewartii subsp. stewartii
    Duong, Duy An; Stevens, Ann M. (PeerJ, 2017-12-06)
    Pantoea stewartii subsp. stewartii is a Gram-negative proteobacterium that causes leaf blight and Stewart’s wilt disease in corn. Quorum sensing (QS) controls bacterial exopolysaccharide production that blocks water transport in the plant xylem at high bacterial densities during the later stage of the infection, resulting in wilt. At low cell density the key master QS regulator in P. stewartii, EsaR, directly represses rcsA, encoding an activator of capsule biosynthesis genes, but activates lrhA, encoding a transcription factor that regulates surface motility. Both RcsA and LrhA have been shown to play a role in plant virulence. In this study, additional information about the downstream targets of LrhA and its interaction with RcsA was determined. A transcriptional fusion assay revealed autorepression of LrhA in P. stewartii and electrophoretic mobility shift assays (EMSA) using purified LrhA confirmed that LrhA binds to its own promoter. In addition, LrhA binds to the promoter for the RcsA gene, as well as those for putative fimbrial subunits and biosurfactant production enzymes in P. stewartii, but not to the flhDC promoter, which is the main direct target of LrhA in Escherichia coli. This work led to a reexamination of the physiological function of RcsA in P. stewartii and the discovery that it also plays a role in surface motility. These findings are broadening our understanding of the coordinated regulatory cascades utilized in the phytopathogen P. stewartii.