• Title/Summary/Keyword: 16S-rRNA

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Biocontrol Characteristics of Bacillus Species in Suppressing Stem Rot of Grafted Cactus Caused by Bipolaris cactivora

  • Bae, Sooil;Kim, Sang Gyu;Kim, Young Ho
    • The Plant Pathology Journal
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    • v.29 no.1
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    • pp.42-51
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    • 2013
  • One of the most important limiting factors for the production of the grafted cactus in Korea is the qualitative and quantitative yield loss derived from stem rots especially caused by Bipolaris cactivora. This study is aimed to develop microbial control agents useful for the control of the bipolaris stem rot. Two bacteria (GA1-23 and GA4-4) selected out of 943 microbial isolates because of their strong antibiotic activity against B. cactivora were identified as Bacillus subtilis and B. amyloliquefaciens, respectively, by the cultural characteristics, Biolog program and 16S rRNA sequencing analyses. Both bacterial isolates significantly inhibited the conidial germination and mycelial growth of the pathogen with no significant difference between the two, of which the inhibitory efficacies varied depending on the cultural conditions such as temperature, nutritional compositions and concentrations. Light and electron microscopy of the pathogen treated with the bacterial isolates showed the inhibition of spore germination with initial malformation of germ tubes and later formation of circle-like vesicles with no hyphal growth and hyphal disruption sometimes accompanied by hyphal swellings and shrinkages adjacent to the bacteria, suggesting their antibiotic mode of antagonistic activity. Control efficacy of B. subtilis GA1-23 and B. amyloliquefaciens GA4-4 on the cactus stem rot were not as high as but comparable to that of fungicide difenoconazole when they were treated simultaneously at the time of pathogen inoculation. All of these results suggest the two bacterial isolates have a good potential to be developed as biocontrol agents for the bipolaris stem rot of the grafted cactus.

Biological Control of Apple Ring Rot on Fruit by Bacillus amyloliquefaciens 9001

  • Li, Yan;Han, Li-Rong;Zhang, Yuanyuan;Fu, Xuechi;Chen, Xinyi;Zhang, Lixia;Mei, Ruhong;Wang, Qi
    • The Plant Pathology Journal
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    • v.29 no.2
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    • pp.168-173
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    • 2013
  • Apple ring rot disease, caused by Botryosphaeria dothidea (Moug. ex. Fr) Ces. et de Not., is one of the most important diseases on apple fruits. In this study, strain 9001 isolated from healthy apple fruits from an infested orchard was evaluated for its biocontrol activity against apple ring rot in vitro and in vivo. Strain 9001 showed obvious antagonistic activity to B. dothidea YL-1 when plated on potato dextrose agar. Soaking healthy apples in the bacterial suspensions of strain 9001 prior to artificial inoculation of fungal pathogen resulted in a dramatic decrease in disease incidence when compared to the control. Moreover, either field application in the growth season or postharvest treatment of apples from infected orchards with bacterial suspensions of strain 9001 resulted in significantly reduced disease incidence within the storage period for 4 months at room temperature. Based on the phylogenetic analysis of 16S rRNA and the gyrA gene, strain 9001 was identified as Bacillus amyloliquefaciens. These results indicated that B. amyloliquefaciens 9001 could be a promising agent in biocontrol of apple ring rot on fruit, which might help to minimize the yield loss of apple fruit during the long postharvest period.

Biocontrol of Late Blight (Phytophthora capsici) Disease and Growth Promotion of Pepper by Burkholderia cepacia MPC-7

  • Sopheareth, Mao;Chan, Sarun;Naing, Kyaw Wai;Lee, Yong Seong;Hyun, Hae Nam;Kim, Young Cheol;Kim, Kil Yong
    • The Plant Pathology Journal
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    • v.29 no.1
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    • pp.67-76
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    • 2013
  • A chitinolytic bacterial strain having strong antifungal activity was isolated and identified as Burkholderia cepacia MPC-7 based on 16S rRNA gene analysis. MPC-7 solubilized insoluble phosphorous in hydroxyapatite agar media. It produced gluconic acid and 2-keto-gluconic acid related to the decrease in pH of broth culture. The antagonist produced benzoic acid (BA) and phenylacetic acid (PA). The authentic compounds, BA and PA, showed a broad spectrum of antimicrobial activity against yeast, several bacterial and fungal pathogens in vitro. To demonstrate the biocontrol efficiency of MPC-7 on late blight disease caused by Phyto-phthora capsici, pepper plants in pot trials were treated with modified medium only (M), M plus zoospore inoculation (MP), MPC-7 cultured broth (B) and B plus zoospore inoculation (BP). With the sudden increase in root mortality, plants in MP wilted as early as five days after pathogen inoculation. However, plant in BP did not show any symptom of wilting until five days. Root mortality in BP was markedly reduced for as much as 50%. Plants in B had higher dry weight, P concentration in root, and larger leaf area compared to those in M and MP. These results suggested that B. cepacia MPC-7 should be considered as a candidate for the biological fertilizer as well as antimicrobial agent for pepper plants.

Isolation and molecular identification of Vibrio harveyi from the look down fish (Selene vomer)

  • Jun, Jin-Woo;Lee, Young-Ran;Giri, Sib Sankar;Kim, Hyoun-Joong;Yun, Sae-Kil;Chi, Cheng;Kim, Sang-Guen;Koh, Jeong-Rack;Jung, Ji-Yun;Lee, Byeong-Chun;Park, Se-Chang
    • Korean Journal of Veterinary Research
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    • v.56 no.2
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    • pp.125-127
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    • 2016
  • Gradual mortality of look down fish (Selene vomer) was observed in a private aquarium in Seoul, showing abnormal swimming behavior and lethargy. A bacterial pathogen from kidney was cultured, identified, and confirmed as Vibrio harveyi using Vitek System 2 and 16S rRNA gene sequencing. A predominant bacterial strain, SNUVh-LW2 was proved to be most closely related to isolates from China by phylogenetic analysis with minimum evolution method. Also, tetracycline was considered as the most sensitive antibiotic agent via antibiotic susceptibility test. The group of fish was treated according to the diagnostic result and no more mortality was observed.

Optimizing the fermentation condition of low salted squid jeotgal by lactic acid bacteria with enhanced antioxidant activity

  • Akther, Fahima;Le, Bao;Chung, Gyuhwa;Yang, Seung Hwan
    • Journal of Applied Biological Chemistry
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    • v.60 no.4
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    • pp.391-402
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    • 2017
  • Lactic acid bacteria (LAB) are widely used as starter culture in food fermentation due to their harmless entity and health beneficial properties along with the ability to change texture, aroma, flavor and acidity of food products. In this study, five different LAB (FB003, FB058, FB077, FB081, and FB111) isolated from different Korean traditional fermented foods, assigned to Lactobacillus plantarum, Pediococcus pentosaceus, Weissella viridescens, Lactobacillus sakei, and Leuconostoc mesenteroides, respectively, on the basis of their physiological properties and 16S rRNA sequence analysis, to use as fermentation starter and check their ability to fasten the ripening time as well as the overall optimization in the fermentation condition. To check their suitability as starters, their safety, acid and bile tolerance, NaCl and temperature resistance, susceptibility to common antibiotics, and antimicrobial activities were determined. Squid jeotgal samples were prepared by adding $10^8CFU/g$ of each strain in different samples, which were then kept for fermentation at $4^{\circ}C$ and checked for their antioxidant activities at 0, 7, 15, and 21-day intervals. The samples fermented with FB003 and FB077 displayed the highest antioxidant activity. This study revealed two effective starter cultures (FB003, FB077) for squid jeotgal fermentation, which presented increased functionalities. The results of this study will lead to the development of novel industrial-scale production avenues for jeotgal preparation, and offer new insights into the prevention and control of chronic diseases.

Optimization of \beta-mammanase Production from Bacillus subtilis JS-1. (\beta-Mannanase를 생산하는 Bacillus subtilis JS-1의 분리 및 효소 생산성)

  • 임지수;정진우;이종수;강대경;강하근
    • Microbiology and Biotechnology Letters
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    • v.31 no.1
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    • pp.57-62
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    • 2003
  • A bacteria strain producing extracellular $\beta$-mannanase was isolated from soil and was identified as Bacillus subtilis by 16S rRNA sequence comparison and biochemical determinations. The optimum pH and temperature for the $\beta$-mannanase activity were 5.0 and 5.5$^{\circ}C$, respectively. The zymogram technique revealed a single protein band exhibiting $\beta$-mannanase activity from the culture supernatant. The molecular mass of the enzyme was estimated at approximately 130 kDa. The addition of 0.5% lactose or 0.5% locust bean gum to the LB medium caused to Increase significantly the $\beta$-mannanase productivity from Bacillus subtilis JS-1. The cells grown on LB medium supplemented with lactose produced maximal enzyme activity at the stationary phase. In contrast to this, the $\beta$-mannanase was induced at the logarithmic phase from the cells grown on LB medium supplemented with locust bean gum. The discrepancy in induction times suggests that $\beta$-mannanase was induced by different induction mechanisms depending on the carbon sources in Bacillus subtilis JS-1 .

Validation of Morphology-based Identification of Two Cynoglossidae Larvae using Mitochondrial DNA (참서대과(Pisces: Cynoglossidae) 자어 2종의 미토콘드리아 DNA에 의한 형태동정의 타당성)

  • Kwun, Hyuck-Joon;Kim, Jin-Koo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.43 no.5
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    • pp.482-488
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    • 2010
  • Three specimens of Cynoglossidae larvae were collected from the southern Korean Sea in May and August of 2009, and were identified using morphological and molecular analysis. Specimens were divided into two groups based on the number of elongated dorsal fin rays on the top of the head: Cynoglossidae sp. A was defined as having two elongated dorsal fin rays, while Cynoglossidae sp. B possessed a single elongated dorsal fin ray. One specimen of Cynoglossidae sp. A, a post-larva with a notochord length (NL) of 5.8 mm was thought to be a Cynoglossus joyneri larva based on the presence of 115 dorsal pterogiophores, 85 anal pterogiophores, and 50 myomeres. Two specimens of Cynoglossidae sp. B, a 4.1 mm NL larva and a 11.3 mm NL juvenile, were thought to be Cynoglossus abbreviatus based on the presence of yolk in the former and 133 dorsal fin rays, 105 anal fin rays, and 63 myomeres in the latter. To test this morphology-based identification, molecular analysis was conducted using 419-422 bp of mitochondrial DNA 16S rRNA. Cynoglossidae sp. A was clearly matched to a Cynoglossus joyneri adult (d=0.000) and Cynoglossidae sp. B clustered closely with Cynoglossus abbreviatus adults (d=0.002). A neighbor-joining tree supported this robust relationship (bootstrap value=100%). Therefore, these molecular data validate the morphological identification of the two Cynoglossidae larval species.

Association between immunoglobulin G1 against Tannerella forsythia and reduction in the loss of attachment tissue

  • Ardila, Carlos Martin;Olarte-Sossa, Mariana;Guzman, Isabel Cristina
    • Journal of Periodontal and Implant Science
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    • v.44 no.6
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    • pp.274-279
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    • 2014
  • Purpose: To evaluate whether the levels of immunoglobulin G (IgG) antibody to Tanerella forsythia are associated with periodontal status. Methods: Patients with a diagnosis of chronic periodontitis were considered candidates for the study; thus 80 chronic periodontitis patients and 28 healthy persons (control group) were invited to participate in this investigation. The presence of T. forsythia was detected by polymerase chain reaction (PCR) analysis using primers designed to target the respective 16S rRNA gene sequences. Peripheral blood was collected from each subject to identify the IgG1 and IgG2 serum antibodies against T. forsythia. All microbiological and immunological laboratory processes were completed blindly, without awareness of the clinical status of the study patients or of the periodontal sites tested. Results: The bivariate analysis showed that lower mean levels of clinical attachment level (CAL) and probing depth were found in the presence of the IgG1 antibody titers against whole-cell T. forsythia; however, only the difference in CAL was statistically significant. In the presence of the IgG2 antibody titers against whole-cell T. forsythia, the periodontal parameters evaluated were higher but they did not show statistical differences, except for plaque. The unadjusted linear regression model showed that the IgG1 antibody against whole-cell T. forsythia in periodontitis patients was associated with a lower mean CAL (${\beta}=-0.654$; 95% confidence interval [CI], -1.27 to -0.28; P<0.05). This statistically significant association remained after adjusting for possible confounders (${\beta}=-0.655$; 95% CI, -1.28 to -0.29; P<0.05). On the other hand, smoking was a statistically significant risk factor in the model (${\beta}=0.704$; 95% CI, 0.24 to 1.38; P<0.05). Conclusions: Significantly lower mean levels of CAL were shown in the presence of the IgG1 antibody titers against whole-cell T. forsythia in periodontitis patients. Thus, the results of this study suggest that IgG1 antibody to T. forsythia may have been a protective factor from periodontitis in this sample.

Conjugated linoleic acid producing potential of lactobacilli isolated from goat (AXB) rumen fluid samples

  • Tyagi, Amrish Kumar;Kumar, Sachin;Choudhury, Prasanta Kumar;Tyagi, Bhawna;Tyagi, Nitin
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.8
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    • pp.1233-1241
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    • 2020
  • Objective: The present investigation was aimed to explore the potential of lactobacilli for conjugated linoleic acid (CLA) production, isolated from rumen fluid samples of lactating goats. Methods: A total of 64 isolates of lactobacilli were obtained using deMan-Rogosa-Sharpe (MRS) agar from rumen fluid of goats and further subjected to morphological and biochemical characterizations. Isolates found as gram-positive, catalase negative rods were presumptively identified as Lactobacillus species and further confirmed by genus specific polymerase chain reaction (PCR). The phylogenetic tree was constructed from the nucleotide sequences using MEGA6. Results: Out of the 64 isolates, 23 isolates were observed positive for CLA production by linoleate isomerase gene-based amplification and quantitatively by UV-spectrophotometric assay for the conversion of linoleic acid to CLA as well as gas chromatography-based assay. In all Lactobacillus species cis9, trans11 isomer was observed as the most predominant CLA isomer. These positive isolates were identified by 16S rRNA gene-based PCR sequencing and identified to be different species of L. ingluviei (2), L.salivarius (2), L. curvatus (15), and L. sakei (4). Conclusion: The findings of the present study concluded that lactic acid bacteria isolated from ruminal fluid samples of goat have the potential to produce bioactive CLA and may be applied as a direct fed microbial to enhance the nutraceutical value of animal food products.

PREVALENCE OF BLACK-PIGMENTED BACTERIA IN INFECTED ROOT CANALS IN KOREA (감염 근관의 흑색세균의 동정)

  • Chung, Ki-Soo;Lim, Sung-Sam;Bae, Kwang-Shik
    • Restorative Dentistry and Endodontics
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    • v.24 no.3
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    • pp.447-452
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    • 1999
  • The role of bacteria in root canals and periapical infections is well known and established. In these bacteria, black-pigmented bacteria(BPH) play important role in endodontic infection. BPB are Gram negative anaerobic rods which are closely related 50 clinical symptoms such as pain, percussion, tenderness, foul odor, etc. In America and Europe, many studies on BPB have been done and are continued. But, relatively few studies have been done in Korea, especially its prevalence in Korean population is not yet studied. The purpose of this study is to establish prevalence of BPB in infected root canals and periapical abscesses in Korean people. Microbial samples were collected from the root canals of 34 intact tooth with periapical rarefactions of endodontic origin and 3 periapical abscesses. All samples were incubated in an anaerobic chamber(Coy, Model No. 77. Ann Arbor, Michigan, USA.). Identification of In microorganism was based on its growth in the anaerobic chamber, colonial pigmentation, colonial morphology, Gram stain, and Rapid ID32A(BioMericux SA/69280 Marcy-l'Etoile/France) results. In addition, the polyme ase chain reaction using specific primers for 16S rRNA genes were used differentiate Prevotella nigrescens for Prevotella intermedia. The results were as follows : 1. In this study, thirteen (35%) of thirty seven samples were positive for the growth of BPB. In thirteen samples, sixteen strains of BPR were found. 2. The most frequently identified BPB in root canals and abscesses in Korean were P. nigrescens 5/37(14%) and P. intermedia 5/37(14%). Porphyromonas gingivalis 3/37(8%), Porphyromonas endodontalis 2/37(5%) and Prevotella loecheii 1/37(3%) were also found. 3. In this study, no significant differences were found between the prevalence of BPB in Korean and that of American and European.

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