• The Plant Pathology Journal
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• 제32권3호
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• pp.266-272
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• 2016

A bacterial tyrosine sulfotransferase, RaxST, is required for activation of rice XA21-mediated immunity, and it catalyzes sulfation of tyrosine residues of Omp1X and RaxX in Xanthomonas oryzae pv. oryzae, a causal agent of bacterial blight in rice. Although RaxST is biochemically well-characterized, biological functions of tyrosine sulfation have not been fully elucidated. We compared protein expression patterns between the wildtype and a raxST knockout mutant using shotgun proteomic analysis. Forty nine proteins displayed a more than 1.5-fold difference in their expression between the wildtype and the mutant strains. Clusters of orthologous groups analysis revealed that proteins involved in cell motility were most abundant, and phenotypic observation also showed that the twitching motility of the mutant was dramatically changed. These results indicate that tyrosine sulfation by RaxST is essential for Xoo movement, and they provide new insights into the biological roles of RaxST in cellular processes.

• The Plant Pathology Journal
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• 제35권5호
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• pp.445-458
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• 2019

The lipopolysaccharide (LPS) composed of lipid A, core, and O-antigen is the fundamental constituent of the outer membrane in gram-negative bacteria. This study was conducted to investigate the roles of LPS in Burkholderia glumae, the phytopathogen causing bacterial panicle blight and seedling rot in rice. To study the roles of the core oligosaccharide (OS) and the O-antigen region, mutant strains targeting the waaC and the wbiFGHI genes were generated. The LPS profile was greatly affected by disruption of the waaC gene and slight reductions were observed in the O-antigen region following wbiFGHI deletions. The results indicated that disruption in the core OS biosynthesis-related gene, waaC, was associated with increased sensitivity to environmental stress conditions including acidic, osmotic, saline, and detergent stress, and to polymyxin B. Moreover, significant impairment in the swimming and swarming motility and attenuation of bacterial virulence to rice were also observed in the waaC-defective mutant. The motility and virulence of O-antigen mutants defective in any gene of the wbiFGHI operon, were not significantly different from the wild-type except in slight decrease in swimming and swarming motility with wbiH deletion. Altogether, the results of present study indicated that the LPS, particularly the core OS region, is required for tolerance to environmental stress and full virulence in B. glumae. To our knowledge, this is the first functional study of LPS in a plant pathogenic Burkholderia sp. and presents a step forward toward full understanding of B. glumae pathogenesis.

• 한국미생물·생명공학회지
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• 제32권1호
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• pp.1-10
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• 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.

• 대한기계학회:학술대회논문집
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• 대한기계학회 2008년도 추계학술대회A
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• pp.1433-1436
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• 2008

Flagellate bacteria such as Escherichia coli or Serratia marcescens possess a remarkable motility system based on a reversible rotary motor. We have employed S. marcescens as microactuators in low Reynolds number fluidic environments to move a larger engineering element around. Microstructures fabricated using conventional microfabrication techniques are blotted on the swarm plate, which leaves a bacterial monolayer on the surface of the microstructure. We have investigated microstructures powered by bacteria to determine how cell orientation on the microstructure surface relates to the swarming patterns as well as how the orientation is affected by the blotting process. This study will help to refine directional control of bacterial transporters by exploiting bacterial sensory mechanisms.

• The Plant Pathology Journal
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• 제34권5호
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• pp.412-425
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• 2018

The Hfq protein is a global small RNA chaperone that interacts with regulatory bacterial small RNAs (sRNA) and plays a role in the post-transcriptional regulation of gene expression. The roles of Hfq in the virulence and pathogenicity of several infectious bacteria have been reported. This study was conducted to elucidate the functions of two hfq genes in Burkholderia glumae, a causal agent of rice grain rot. Therefore, mutant strains of the rice-pathogenic B. glumae BGR1, targeting each of the two hfq genes, as well as the double defective mutant were constructed and tested for several phenotypic characteristics. Bacterial swarming motility, toxoflavin production, virulence in rice, siderophore production, sensitivity to $H_2O_2$, and lipase production assays were conducted to compare the mutant strains with the wild-type B. glumae BGR1 and complementation strains. The hfq1 gene showed more influence on bacterial motility and toxoflavin production than the hfq2 gene. Both genes were involved in the full virulence of B. glumae in rice plants. Other biochemical characteristics such as siderophore production and sensitivity to $H_2O_2$ induced oxidative stress were also found to be regulated by the hfq1 gene. However, lipase activity was shown to be unassociated with both tested genes. To the best of our knowledge, this is the first study to elucidate the functions of two hfq genes in B. glumae. Identification of virulence-related factors in B. glumae will facilitate the development of efficient control measures.

• The Plant Pathology Journal
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• 제29권3호
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• pp.249-259
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• 2013

Burkholderia glumae causes rice grain rot and sheath rot by producing toxoflavin, the expression of which is regulated by quorum sensing (QS). The QS systems of B. glumae rely on N-octanoyl homoserine lactone, synthesized by TofI and its cognate receptor TofR, to activate the genes for toxoflavin biosynthesis and an IclR-type transcriptional regulator gene, qsmR. To understand genome-wide transcriptional profiling of QS signaling, we employed RNAseq of the wild-type B. glumae BGR1 with QS-defective mutant, BGS2 (BGR1 tofI::${\Omega}$) and QS-dependent transcriptional regulator mutant, BGS9 (BGR1 qsmR::${\Omega}$). A comparison of gene expression profiling among the wild-type BGR1 and the two mutants before and after QS onset as well as gene ontology (GO) enrichment analysis from differential expressed genes (DEGs) revealed that genes involved in motility were highly enriched in TofI-dependent DEGs, whereas genes for transport and DNA polymerase were highly enriched in QsmR-dependent DEGs. Further, a combination of pathways with these DEGs and phenotype analysis of mutants pointed to a couple of metabolic processes, which are dependent on QS in B. glumae, that were directly or indirectly related with bacterial motility. The consistency of observed bacterial phenotypes with GOs or metabolic pathways in QS-regulated genes implied that integration RNAseq with GO enrichment or pathways would be useful to study bacterial physiology and phenotypes.

• 한국미생물·생명공학회지
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• 제49권4호
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• pp.587-593
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• 2021

Infertility is a key issue affecting mood and behavior in men. Microorganisms are one of the primary etiological agents that may be associated with infertility. The objective of the present study was to identify bacterial causative agents from the semen of infertile subjects and determine the effect of bacterial infection on sperm quality, as well as determine the susceptibility of these bacteria to drugs. Forty semen samples from 30 infertile patients and 10 fertile individuals were collected. The pH, volume, motility, and concentration of semen were analyzed. The samples were processed and identified by biochemical testing using API identification kits. The antibiotic susceptibility pattern was determined using the disc diffusion method. Abnormal sperm quality was observed. The mean age of the individual and their sperm morphology, concentration, progressive motility, pH level, and pus cell content were 31.9 years, 2.7%, 10.4 million/ml, 27.3%, 8.3, and 5.7, respectively. Among the tested samples, oligoasthenozoospermia was found to show the highest occurrence, at 27/30 samples, followed by teratozoospermia, at 25/30 samples, and asthenozoospermia, at 22/30 samples. Of the tested infertile patients' sperm, 19, 6, and 5 isolates were identified as Escherichia coli, Klebsiella pneumonia, and Staphylococcus epidermidis, respectively. The results also revealed multi-drug resistance in the bacteria. Compared to that shown by the other tested antibiotics, amikacin showed higher activity against all isolated bacteria. However, the bacteria exhibited maximum resistance against gentamicin, cefotaxime, levofloxacin, and ampicillin. In conclusion, leukocytospermia and bacterial infections are possibly responsible for sperm abnormalities. Multi-drug resistant bacteria were detected. Gentamicin, cefotaxime, levofloxacin and ampicillin were shown the highest resistance, while amikacin was the most effective antimicrobial agent against the isolated bacteria.

• The Plant Pathology Journal
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• 제39권1호
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• pp.123-135
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• 2023

Previously, Pseudomonas plecoglossicida YJR13 and Pseudomonas putida YJR92 from a sequential screening procedure were proven to effectively control Phytophthora blight caused by Phytophthora capsici. In this study, we further investigated the anti-oomycete activities of these strains against mycelial growth, zoospore germination, and germ tube elongation of P. capsici. We also investigated root colonization ability of the bacterial strains in square dishes, including cell motility (swimming and swarming motilities) and biofilm formation. Both strains significantly inhibited mycelial growth in liquid and solid V8 juice media and M9 minimal media, zoospore germination, and germ tube elongation compared with Bacillus vallismortis EXTN-1 (positive biocontrol strain), Sphingomonas aquatilis KU408 (negative biocontrol strain), and MgSO₄ solution (untreated control). In diluted (nutrient-deficient) V₈ juice broth, the tested strain populations were maintained at >10⁸ cells/ml, simultaneously providing mycelial inhibitory activity. Additionally, these strains colonized pepper roots at a 10⁶ cells/ml concentration for 7 days. The root colonization of the strains was supported by strong swimming and swarming activities, biofilm formation, and chemotactic activity towards exudate components (amino acids, organic acids, and sugars) of pepper roots. Collectively, these results suggest that strains YJR13 and YJR92 can effectively suppress Phytophthora blight of pepper through direct anti-oomycete activities against mycelial growth, zoospore germination and germ tube elongation. Bacterial colonization of pepper roots may be mediated by cell motility and biofilm formation together with chemotaxis to root exudates.

• 생명과학회지
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• 제21권10호
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• pp.1487-1493
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• 2011

염농도에 따른 E. tarda CK41의 운동성을 알아보기 위하여 1.0%와 3.5%의 염농도를 가지는 운동성 측정 배지에서 집락의 변화를 관찰한 결과, 3.5% 염농도 조건에서 운동성이 감소하는 것을 확인할 수 있었다. 1.0%과 3.5% 염농도 조건에서의 생육도를 측정해본 결과 각 염농도 조건에 따른 생균수의 차이는 매우 적은 것으로 보아, 높은 염농도에서의 운동성의 감소는 생육정체가 아닌 실질적인 운동성의 차이에 의함을 알 수 있었다. 이러한 염농도에 의한 운동성의 차이가 편모에 의한 것인지를 알아보기 위하여 투과 전자 현미경으로 형태학적 관찰을 해본 결과, 3.5% 염농도에서는 편모의 형성이 되지 않음을 확인하였다. E. tarda는 PFAD와 FDP 두개의 편모 유전자를 가지며 이들간의 아미노산 상동률은 93%로 높은 편이다. 편모의 발현양의 확인을 위하여 PFAD 특이적인 다클론성 항체를 제작하기 위하여, PFAD를 과발현시키는 재조합 플라스미드 pBP793을 구축하여 대장균 발현시스템으로 발현시켜 정제한 후, 토끼에서 면역반응을 유도하여 특이 항체를 제작하였다. PFAD 특이적인 다클론성 항체를 이용한 immunoblot assay 결과, 3.5% 염농도 조건에서 배양한 E. tarda CK41의 경우 1.0% 염농도에서 보다 반응하는 면역 활성 단백질 밴드가 낮은 것으로 측정되었다. 이러한 결과를 종합하여 볼 때, 염농도가 높은 해수환경에서의 운동성의 감소는 E. tarda CK41의 편모 단백질이 제대로 발현되지 않아 기능적인 편모의 형성이 이루어지지 않는다는 것을 예증하고 있다. 향후 연구에서 어떠한 메카니즘에 의해 염농도가 flagellin의 발현을 조절하는지를 밝힐 필요가 있다.

• 대한치과의료관리학회지
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• 제6권1호
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• pp.28-35
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• 2018

Halitosis is primarily caused by bacterial decay. The bacteria, which originate from biofilms such as dental plaque, show abnormal proliferation due to dental caries, periodontal diseases, soft tissue infections, and tongue diseases. Most studies on halitosis have exclusively focused on gram-negative bacteria in the oral cavity rather than on general oral microorganisms including oral fungi. This study analyzed oral fungal hyphae, as well as distribution and motility of oral microorganisms, and provided basic data on the control of halitosis. Our results revealed that the greater is the number of cocci bacteria, the higher is the halitosis value, or bad breath value (BBV), suggesting that cocci have a strongly positive correlation with halitosis (r=0.379, p=0.030). Moreover, there was no significant difference in the morphology or distribution of motile bacteria and motility score, with respect to BBV. Lastly, we investigated the relationship between halitosis and oral fungal hyphae. We found that a higher BBV corresponded with a greater number of fungal hyphae and that patients with fungal hyphae scored a higher BBV. However, this result was not statistically significant. In conclusion, this study provided the preliminary data on oral microorganisms and halitosis, but further studies are needed to analyze the relationship between oral microorganisms and halitosis.