• The Plant Pathology Journal
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• v.24 no.2
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• pp.143-151
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• 2008

To define the functions of the rpf genes in Xanthomonas oryzae pv. oryzae (Xoo), which regulates pathogenicity factors in Xanthomonas campestris pv. campestris (Xcc), marker-exchange mutants of each rpf gene were generated. When the mutants were inoculated on a susceptible cultivar, the lesion lengths caused by the rpfB, rpfC, rpfF, and rpfG mutants were significantly smaller than those caused by the wild type, whereas those caused by the rpfA, rpfD, and rpfI mutants were not. Several virulence determinants, including extracellular polysaccharide (EPS) production, xylanase production, and motility, were significantly decreased in the four mutants. However, the cellulase activity in the mutants was unchanged. Complementation of the rpfB and rpfC mutations restored the virulence and the expression of the virulence determinants. Expression analysis of 14 virulence genes revealed that the expression of genes related to EPS production (gumG and gumM), LPS (xanA, xanB, wxoD, and wxoC), phytase (phyA), xylanase (xynB), lipase (lipA), and motility (pitA) were reduced significantly in the mutants rpfB, rpfC, rpfF, and rpfG. In contrast, the expression of genes related to cellulase (eglxob, clsA), cellobiosidase (cbsA), and iron metabolism (fur) was unchanged. The results of this study clearly show that rpfB, rpfC, rpfF, and rpfG are important for the virulence of Xoo KACC10859, and that virulence genes are regulated differently by the Rpfs.

• Food Science and Biotechnology
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• v.16 no.5
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• pp.778-782
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• 2007

This study assessed the effects of the antimicrobial substances sulforaphane, grapefruit seed extracts (GSE), and reuterin on the expression of Salmonella HilA and InvF virulence gene using a LacZY assay (${\beta}$-galactosidase assay) with hilA:lacZY and invF:lacZY fusion strains of Salmonella typhimurium SL1344. Salmonella was grown for 8 hr at $37^{\circ}C$ in the presence of diluted antimicrobial substances ($2\;{\mu}g/mL$ sulforaphane, $20\{\mu}g/mL$ GSE, and 0.26 mM reuterin) at concentrations that did not inhibit the cellular growth of Salmonella. Sulforaphane inhibited the expression of HilA and InvF by 50-90 and 20-80%, respectively. GSE also inhibited the expression of both genes, but to a lesser degree. Among the 3 antimicrobial substances, reuterin showed the least inhibition, which was abolished after 3-4 hr. None of the antimicrobial substances inhibited the ${\beta}$-galactosidase enzyme activity of S. typhimurium. The assay used in this study represents a very sensitive method for screening bioactive substances that inhibit the expression of virulence genes in Salmonella.

• The Plant Pathology Journal
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• v.36 no.5
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• pp.450-458
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• 2020

Pepper and tomato plants infected with two Clavibacter species, C. capsici and C. michiganensis have shown different patterns of disease development depending on their virulence. Here, we investigated how pepper and tomato plants respond to infection by the high-virulent or low-virulent Clavibacter strains. For this, we chose two strains of each Clavibacter species to show different virulence level in the host plants. Although low-virulent strains showed less disease symptoms, they grew almost the same level as the high-virulent strains in both plants. To further examine the response of host plants to Clavibacter infection, we analyzed the expression patterns of plant defense-related genes in the leaves inoculated with different strains of C. capsici and C. michiganensis. Pepper plants infected with high-virulent C. capsici strain highly induced the expression of CaPR1, CaDEF, CaPR4b, CaPR10, and CaLOX1 at 5 days after inoculation (dai), but their expression was much less in low-virulent Clavibacter infection. Expression of CaSAR8.2 was induced at 2 dai, regardless of virulence level. Expression of GluA, Pin2, and PR2 in tomato plants infected with high-virulent C. michiganensis were much higher at 5 dai, compared with mock or low-virulent strain. Expression of PR1a, Osmotin-like, Chitinase, and Chitinase class 2 was increased, regardless of virulence level. Expression of LoxA gene was not affected by Clavibacter inoculation. These results suggested that Clavibacter infection promotes induction of certain defense-related genes in host plants and that differential expression of those genes by low-virulent Clavibacter infection might be affected by their endophytic lifestyle in plants.

• Korean Journal of Microbiology
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• v.54 no.1
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• pp.1-8
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• 2018

A prokaryotic cell has various histone-like proteins also known as nucleoid-associated proteins (NAPs). These proteins bind AT-rich sequence at DNA, which induce DNA wrapping, bending, and bridging, and subsequently regulate the gene expression in bacteria. Because NAPs function in transcriptional silencing of virulence genes, it is important to study their roles in gene silencing and specific mechanisms of these proteins. In this review, we discussed two well-known NAPs, H-NS, and HU, and summarized their roles for gene expression in Escherichia coli and Salmonella Typhimurium. Through the oligomerization and filamentation of H-NS, it represses the expression of virulence genes in human pathogenic bacteria, such as Salmonella Typhimurium, and it works with other NAPs positively or negatively. Recently, H-NS also regulates typhoid toxin expression, which causes typhoid fever and systemic disease in human. Additionally, HU regulates the expression of genes related to both virulence and physiology of Salmonella. Therefore, we suggest that NAPs like H-NS and HU are crucial factors to reveal the molecular mechanisms of virulence gene expression in bacteria.

• Journal of Microbiology and Biotechnology
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• v.19 no.12
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• pp.1520-1526
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• 2009

We have identified the iscR (PA3815) gene encoding an iron-sulfur cluster assembly regulator homolog as one of the genes required for peroxide resistance in Pseudomonas aeruginosa PA14. Here, we present the phenotypic characterization of an iscR deletion mutant in terms of KatA expression, stress responses, and virulence. The iscR null mutant exhibited reduced KatA activity at the posttranslational level, hypersensitivity to hydrogen peroxide, and virulence-attenuation in Drosophila melanogaster and mouse peritonitis models. These phenotypes were fully restored by multicopy-based expression of katA. These results suggest that the requirement of IscR in P. aeruginosa is related to the proper activity of KatA, which is crucial for peroxide resistance and full virulence of this bacterium.

• Journal of Microbiology and Biotechnology
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• v.14 no.2
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• pp.367-372
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• 2004

Infection of Drosophila melanogaster adults with Pseudomonas aeruginosa (PA14) can kill the flies within 48h. We found that the virulence of PA14 was significantly attenuated when treated with a reducing agent, dithiothreitol (DTT), prior to infection. Infection with DTT-treated PA14 elevated Metchnikowin expression at 22 h post-infection and the virulence of DTT-treated PA14 was not attenuated in Dif and Relish mutants. These results suggest that DTT pre-treatment of PA14 can aggravate certain virulence factors that may be required to paralyze fly immune responses, triggering Metchnikowin expression via Dif and Relish activations.

• Journal of Microbiology
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• v.38 no.3
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• pp.117-121
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• 2000

Recent advances in studies of bacterial gene expression and light microscopy show that cell-to cell communication and communication and community behavior are the rule rather than the exception. One type of cell-cell communication, quorum sensing in Gram-negative bacteria involves acyl-homoserine lactone signals. This type of quorum sension represents a dedicated communication system that enables a given species to sense when it has reached a critical population density. and to respond by activating expression of specific genes. The LuxR and LuxI proteins of Vibrio fisheri are the founding members of the acyl-homoserine lactone quorum sensing signal receptor and signal generator families of proteins. Acyl-homeserine lactone signaling in Pseudomonas aeruginosa is one model for the relationship between quorum sensing community behavior, and virulence. In the P. aeruginosa model. quorum sensing is required for normal biofilm maturation and virulence. There are multiple quorum-sensing circuits that control the expression of dozens of specific genes in P. aeruginosa.

• Microbiology and Biotechnology Letters
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• v.47 no.4
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• pp.651-661
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• 2019

Targeting the pathogen viability using drugs is associated with development of drug resistance due to selective pressure. Hence, there is an increased interest in developing agents that target bacterial virulence. In this study, the inhibitory effect of ciclopirox, an antifungal agent with iron chelation potential, on the microbial virulence factors was evaluated in 26 clinical MDR Pseudomonas aeruginosa isolates collected from Alexandria Main University Hospital, a tertiary hospital in Egypt. Treatment with 9 ㎍/ml ciclopirox inhibited the hemolytic activity in 70% isolates, reduced pyocyanin production, decreased protease secretion in 46% isolates, lowered twitching and swarming motility, and decreased biofilm formation by 1.5- to 4.5-fold. The quantitative real-time PCR analysis revealed that treatment with ciclopirox downregulated the expression levels of alkaline protease (aprA) and pyocyanin (phzA1). Ciclopirox is used to treat hematological malignancies and the systemic administration of ciclopirox is reported to have adequate oral absorption with a satisfactory drug safety profile. It is important to calculate the appropriate clinical dose and therapeutic index to reposition ciclopirox from a topical antifungal agent to a promising virulence-modifying agent agent against P. aeruginosa, a problematic Gram-negative pathogen.

• Journal of Microbiology and Biotechnology
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• v.19 no.8
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• pp.765-773
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• 2009

Edwardsiella tarda is a pathogen with a broad host range that includes human and animals. The E. tarda hemolysin (Eth) system, which comprises EthA and EthB, is a noted virulence element that is widely distributed in pathogenic isolates of E. tarda. Previous study has shown that the expression of ethB is regulated by iron, which suggests the possibility that the ferric uptake regulator (Fur) is involved in the regulation of ethB. The work presented in this report supports the previous findings and demonstrates that ethB expression was decreased under conditions when the E. tarda Fur ($Fur_{Et}$) was overproduced, and enhanced when $Fur_{Et}$ was inactivated. We also identified a second ethB regulator, EthR, which is a transcription regulator of the GntR family. EthR represses ethB expression by direct interaction with the ethB promoter region. In addition to ethB, EthR also modulates, but positively, luxS expression and AI-2 production by binding to the luxS promoter region. The expression of ethR itself is subject to negative autoregulation; interference with this regulation by overexpressing ethR during the process of infection caused (i) drastic changes in ethB and luxS expressions, (ii) vitiation in the tissue dissemination and survival ability of the bacterium, and (iii) significant attenuation of the overall bacterial virulence. These results not only provide new insights into the regulation mechanisms of the Eth hemolysin and LuxS/AI-2 quorum sensing systems but also highlight the importance of these systems in bacterial virulence.

• International Journal of Oral Biology
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• v.43 no.4
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• pp.171-175
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• 2018

Quorum sensing (QS) is a cell density-dependent communication mechanism between bacteria through small signaling molecules. When the number of QS signaling molecules reaches a threshold, they are transported back into the cells or recognized by membrane-bound receptors, triggering gene expression which affects various phenotypes including bioluminescence, virulence, adhesion, and biofilm formation. These phenotypes are beneficial for bacterial survival in harsh environments. This review summarizes the application of QS inhibitors for control of biofilm formation and virulence expression of periodontal pathogens.