• Journal of Microbiology and Biotechnology
• /
• v.28 no.11
• /
• pp.1896-1907
• /
• 2018

Salmonellosis is commonly associated with meat and poultry products, but an increasing number of Salmonella outbreaks have been attributed to contaminated vegetables and fruits. Enteric pathogens including Salmonella enterica spp. can colonize diverse produce and persist for a long time. Considering that fresh vegetables and fruits are usually consumed raw without heat treatments, Salmonella contamination may subsequently lead to serious human infections. In order to understand the underlying mechanism of Salmonella adaptation to produce, we investigated the transcriptomics of Salmonella in contact with green vegetables, namely cabbage and napa cabbage. Interestingly, Salmonella pathogenicity island (SPI)-1 genes, which are required for Salmonella invasion into host cells, were up-regulated upon contact with vegetables, suggesting that SPI-1 may be implicated in Salmonella colonization of plant tissues as well as animal tissues. Furthermore, Salmonella transcriptomic profiling revealed several genetic loci that showed significant changes in their expression in response to vegetables and were associated with bacterial adaptation to unfavorable niches, including STM14_0818 and STM14_0817 (speF/potE), STM14_0880 (nadA), STM14_1894 to STM14_1892 (fdnGHI), STM14_2006 (ogt), STM14_2269, and STM14_2513 to STM14_2523 (cbi operon). Here, we show that nadA was required for bacterial growth under nutrient-restricted conditions, while the other genes were required for bacterial invasion into host cells. The transcriptomes of Salmonella in contact with cabbage and napa cabbage provided insights into the comprehensive bacterial transcriptional response to produce and also suggested diverse virulence determinants relevant to Salmonella survival and adaptation.

• Korean Journal of Veterinary Research
• /
• v.39 no.2
• /
• pp.327-337
• /
• 1999

The transposon TnphoA was used to generate avirulent mutants from a type A Pasteurella multocida. A suicide vector plasmid pRT733 carrying TnphoA, having the kanamycin resistant gene and harbored in Escherichia coli K-12 strain SM10(${\lambda}pir$), was mated with streptomycin resistant P. multocida P-1059 strain as recipient. This resulted in the generation of two TnphoA insertion mutants (transconjugants, tc95-a and tc95-b) which were resistant both to kanamycin ($Km^{R}$) and streptomycin ($Sm^{R}$), secreted alkaline phosphatase, and were avirulent to turkeys. Southern blot hybridization using two probes derived from internal fragments of TnphoA, confirmed the insertion of TnphoA into 12.9kb or 13.7kb DNA fragment from the EcoRV digested genomic fragments of transconjugants. The two transconjugants, tc95-a and tc95-b, were distinguishable from their parent strains by differences in ribotypes, and outer membrane protein profiles. TnphoA insertion in both transconjugants also resulted in constitutive expression of a 33Kd iron regulated outer membrane protein (IROMP). The gene encoding $Sm^{R}$ was also located within the same 12.9kb EcoRV genomic fragment from both transconjugants. Furthermore, our finding that the recipient P. multocida P-1059 $Sm^{R}$ strain and both transconjugants were avirulent to turkeys suggest that the either 12.9kb or 13.7kb genomic DNA contains the virulence gene and speculate that the presence of $Sm^{R}$ gene or TnphoA insertion may be responsible for regulating and inactivating the gene(s) encoding virulence in P. multocida.

• Journal of Microbiology and Biotechnology
• /
• v.17 no.2
• /
• pp.234-243
• /
• 2007

Vibrio vulnificus is an opportunistic pathogen that causes septicemia in humans. To identify the genes associated with its pathogenicity, in vivo expression technology (IVET) was used to select genes specifically expressed in a host, yet not significantly in vitro. Random lacZ-fusions in the genome of V vulnificus strain MO6-24/O were constructed using an IVET vector, pSG3, which is a suicide vector containing promoterless-aph and -lacZ as reporter genes. A total of ${\sim}18,000$ resulting library clones were then intraperitoneally injected into BALB/c mice using a colony forming unit (CFU) of $1.6{\times}10^6$. Two hours after infection, kanamycin was administered at $200{mu}g$ per gram of mouse weight. After two selection cycles, 11 genes were eventually isolated, which were expressed only in the host. Among these genes, VV20781 and VV21007 exhibiting a homology to a hemagglutinin gene and tolC, respectively, were selected based on having the highest frequency. When compared to wild-type cells, mutants with lesions in these genes showed no difference in the rate of growth rate, yet a significant decrease in cytotoxicity and the capability to form a biofilm.

• Journal of Microbiology and Biotechnology
• /
• v.22 no.7
• /
• pp.889-893
• /
• 2012

Appressorium is a specialized infection structure of filamentous pathogenic fungi and plays an important role in establishing a pathogenic relationship with the host. The Egh16/Egh16H family members are involved in appressorium formation and pathogenesis in pathogenic filamentous fungi. In this study, a homolog of Egh16H, Magas1, was identified from an entomopathogenic fungus, Metarhizium acridum. The Magas1 protein shared a number of conserved motifs with other Egh16/Egh16H family members and specifically expressed during the appressorium development period. Magas1-EGFP fusion expression showed that Magas1 protein was not localized inside the cell. Deletion of the Magas1 gene had no impact on vegetative growth, conidiation and appressorium formation, but resulted in a decreased mortality of host insect when topically inoculated. However, the mortality was not significant between the Magas1 deletion mutant and wild-type treatment when the cuticle was bypassed by injecting conidia directly into the hemocoel. Our results suggested that Magas1 may influence virulence by affecting the penetration of the insects' cuticle.

• The Korean Journal of Mycology
• /
• v.38 no.2
• /
• pp.89-94
• /
• 2010

Plant pathogenic fungi are the most diverse and drastic causal agents of crop diseases threatening stable food production all over the world. Plant have evolved efficient innate immune system to scout and counterattack fungal invasion and pathogenic fungi also developed virulence system to nullify plant resistance machinery or signaling pathways and to propagate and dominate within their niche. A growing body of evidences suggests that post translational modifications (PTMs) and selective/nonselective degradations of proteins involved in virulence expression of plant pathogenic fungi and plant defense machinery should play pivotal roles during the compatible and incompatible interactions. This review elucidates recent investigations about the effects of PTMs and protein degradations on host defense and fungal pathogens' invasions.

• Korean Journal of Food Science and Technology
• /
• v.54 no.2
• /
• pp.241-246
• /
• 2022

Antibiotic resistance is a serious problem to food safety as well as human healthcare. To avoid this, there are several approaches for a new class of antibiotic agents that target only production of virulence factors such as biofilm without bacterial growth defect. The objective of this study was to investigate the antibiofilm activity of tuberostemonine in Staphylococcus aureus. Tuberostemonine significantly reduced the biofilm formation (26.07-47.02%) in the crystal violet assay whereas there were no effect on S. aureus growth. The dispersion in preformed biofilm was also observed by confocal laser scanning microscopy (CLSM). Quantification real-time PCR revealed that the icaA and agrA expression having an important role in biofilm production of S. aureus were strongly affected with tuberostemonine. These results suggest that tuberostemonine has potential for controlling biofilm formation and dispersion by effect on virulence regulation of S. aureus.

• Journal of Microbiology and Biotechnology
• /
• v.34 no.3
• /
• pp.562-569
• /
• 2024

Xanthomonas oryzae pv. oryzae (Xoo) causes a devastating bacterial leaf blight in rice. Here, the antimicrobial effects of ᴰ-limonene, ᴸ-limonene, and its oxidative derivative carveol against Xoo were investigated. We revealed that carveol treatment at ≥ 0.1 mM in liquid culture resulted in significant decrease in Xoo growth rate (> 40%) in a concentration-dependent manner, and over 1 mM, no growth was observed. The treatment with ᴰ-limonene and ᴸ-limonene also inhibited the Xoo growth but to a lesser extent compared to carveol. These results were further elaborated with the assays of motility, biofilm formation and xanthomonadin production. The carveol treatment over 1 mM caused no motilities, basal level of biofilm formation (< 10%), and significantly reduced xanthomonadin production. The biofilm formation after the treatment with two limonene isomers was decreased in a concentration-dependent manner, but the degree of the effect was not comparable to carveol. In addition, there was negligible effect on the xanthomonadin production mediated by the treatment of two limonene isomers. Field emission-scanning electron microscope (FE-SEM) unveiled that all three compounds used in this study cause severe ultrastructural morphological changes in Xoo cells, showing shrinking, shriveling, and holes on their surface. Moreover, quantitative real-time PCR revealed that carveol and ᴰ-limonene treatment significantly down-regulated the expression levels of genes involved in virulence and biofilm formation of Xoo, but not with ᴸ-limonene. Together, we suggest that limonenes and carveol will be the candidates of interest in the development of biological pesticides.

• Journal of Microbiology and Biotechnology
• /
• v.34 no.9
• /
• pp.1757-1768
• /
• 2024

The expression of two major virulence factors of Vibrio cholerae, cholera toxin (CT) and toxin co-regulated pilus (TCP), is induced by environmental stimuli through a cascade of interactions among regulatory proteins known as the ToxR regulon when the bacteria reach the human small intestine. ToxT is produced via the ToxR regulon and acts as the direct transcriptional activator of CT (ctxAB), TCP (tcp gene cluster), and other virulence genes. Unsaturated fatty acids (UFAs) and several small-molecule inhibitors of ToxT have been developed as antivirulence agents against V. cholerae. This study reports the inhibitory effects of fatty acids and virstatin (a small-molecule inhibitor of ToxT) on the transcriptional activation functions of ToxT in isogenic derivatives of V. cholerae strains containing various toxT alleles. The fatty acids and virstatin had discrete effects depending on the ToxT allele (different by 2 amino acids), V. cholerae strain, and culture conditions, indicating that V. cholerae strains could overcome the effects of UFAs and small-molecule inhibitors by acquiring point mutations in toxT. Our results suggest that small-molecule inhibitors should be examined thoroughly against various V. cholerae strains and toxT alleles during development.

• Journal of Photoscience
• /
• v.9 no.2
• /
• pp.323-325
• /
• 2002

Bacillus Anthracis is the causative agent of anthrax. The major virulence factors are a poly-D glutamic acid capsule and three-protein component exotoxin, which is collectively known as anthrax toxin, protective antigen (PA, 83 kDa), lethal factor (LF, 90 kDa), and edema factor (EF, 89 kDa). These three proteins individually have no known toxic activities, but in combination with PA form two toxins (lethal toxin and edema toxin), causing different pathogenic responses in animals and cultured cells. However, it remains to be elucidated for pathogenic mechanism of anthrax toxin. In this study, we constructed toxin component in bacterial overexpression system and purified the native toxin from Bacillus anthracis delta sterne F32 using FPLC system. Recombinant toxin showed high homogeneity and rapid purification processes. Also, this recombinant toxin was comparable to B. anthracis native toxin in terms of cytotoxic effects on cultured cell lines.

• Microbiology and Biotechnology Letters
• /
• v.32 no.1
• /
• pp.1-10
• /
• 2004

Many bacteria monitor their population density and control the expression of specialized gene sets in response to bacterial cell density based on a mechanism referred to as quorum sensing. In all cases, quorum sensing involves the production and detection of extracellular signaling molecules, auto inducers, as which Gram-negative and Gram-positive bacteria use most prevalently acylated homoserine lactones and processed oligo-peptides, respectively. Through quorum-sensing communication circuits, bacteria regulate a diverse array of physiological functions, including virulence, symbiosis, competence, conjugation, antibiotic production, motility, sporulation, and biofilm formation. Many pathogens have evolved quorum-sensing mechanisms to mount population-density-dependent attacks to over-whelm the defense responses of plants, animals, and humans. Since these AHL-mediated signaling mechanisms are widespread and highly conserved in many pathogenic bacteria, the disruption of quorum-sensing system might be an attractive target for novel anti-infective therapy. To control AHL-mediated pathogenicity, several promising strategies to disrupt bacterial quorum sensing have been reported, and several chemicals and enzymes have been also investigated for years. These studies indicate that anti-quorum sensing strategies could be developed as possible alternatives of antibiotics.