• Title/Summary/Keyword: Plant pathogenic bacteria

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A New Approach Using the SYBR Green-Based Real-Time PCR Method for Detection of Soft Rot Pectobacterium odoriferum Associated with Kimchi Cabbage

  • Yong Ju, Jin;Dawon, Jo;Soon-Wo, Kwon;Samnyu, Jee;Jeong-Seon, Kim;Jegadeesh, Raman;Soo-Jin, Kim
    • The Plant Pathology Journal
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    • v.38 no.6
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    • pp.656-664
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    • 2022
  • Pectobacterium odoriferum is the primary causative agent in Kimchi cabbage soft-rot diseases. The pathogenic bacteria Pectobacterium genera are responsible for significant yield losses in crops. However, P. odoriferum shares a vast range of hosts with P. carotovorum, P. versatile, and P. brasiliense, and has similar biochemical, phenotypic, and genetic characteristics to these species. Therefore, it is essential to develop a P. odoriferumspecific diagnostic method for soft-rot disease because of the complicated diagnostic process and management as described above. Therefore, in this study, to select P. odoriferum-specific genes, species-specific genes were selected using the data of the P. odoriferum JK2.1 whole genome and similar bacterial species registered with NCBI. Thereafter, the specificity of the selected gene was tested through blast analysis. We identified novel species-specific genes to detect and quantify targeted P. odoriferum and designed specific primer sets targeting HAD family hydrolases. It was confirmed that the selected primer set formed a specific amplicon of 360 bp only in the DNA of P. odoriferum using 29 Pectobacterium species and related species. Furthermore, the population density of P. odoriferum can be estimated without genomic DNA extraction through SYBR Green-based real-time quantitative PCR using a primer set in plants. As a result, the newly developed diagnostic method enables rapid and accurate diagnosis and continuous monitoring of soft-rot disease in Kimchi cabbage without additional procedures from the plant tissue.

Investigation on Antibacterial and Antioxidant Activities, Phenolic and Flavonoid Contents of Some Thai Edible Plants as an Alternative for Antibiotics

  • Lee, J.H.;Cho, S.;Paik, H.D.;Choi, C.W.;Nam, K.T.;Hwang, S.G.;Kim, Soo-Ki
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.10
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    • pp.1461-1468
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    • 2014
  • This study was aimed to examine the antibacterial and antioxidative properties of seven edible plants from Thailand to develop alternative antibiotics as feed additives. The plants include Citrus aurantifolia Swingle (Lime) fruits and its leaves, Sesbania grandiflora L. (Agati sesbania) leaves, Piper sarmentosum Roxb (Wild betal) leaves, Curcuma domestica Valeton (Turmeric) roots, Morinda citrifolia L. (Beach mulberry) leaves, Cassia siamea britt (Siamea cassia) leaves, and Cocos nucifera L. (Coconut) peels. The plants were extracted by methanol, n-hexane, chloroform, ethyl acetate, butanol and water. Antibacterial activities with minimum inhibitory concentration (MIC) were determined by agar diffusion assay against Escherichia coli, Burkholderia sp., Haemopilus somnus, Haemopilus parasuis, and Clostridium perfringens that were considered pathogenic strains in livestock infection. Methanol extracts of C. aurantifolia Swingle fruits and leaves showed the broadest spectrum of antibacterial activities except for C. perfringens. Butanol extract of S. grandiflora L. leaves showed the strongest activity against Burkholderia sp. with MIC, $135{\mu}g/mL$. P. sarmentosum Roxb leaves showed antibacterial activities against E. coli, Burkholderia sp. and H. parasuis. Ethyl acetate and water extracts from C. domesitca Valeton roots showed MIC of $306{\mu}g/mL$ and $183{\mu}g/mL$, respectively against only C. perfringens. Antioxidative activity was determined by 2-diphenyl-2-picryl hydrazyl photometric assay. The methanol extracts of C. aurantifolia Swingle fruits and P. sarmentosum Roxb leaves showed the highest antioxidant activity among all the extracts with 3.46 mg/mL and 2.70 mg/mL effective concentration 50% ($EC_{50}$) values, respectively. Total contents of phenolics and flavonoids were measured from the plant extracts. Methanol extracts of S. grandiflora L. and chloroform extracts of C. domestica Valeton were found to have the highest amount of total phenolics, 41.7 and $47.8{\mu}g/mL$, respectively. Flavonoid content of methanol extracts in S. grandiflora L. T was $22.5{\mu}g/mL$ and the highest among plant extracts tested. These results indicated that C. aurantifolia Swingle, S. grandiflora L., P. sarmentosum Roxb, and C. domestica Valeton have antibacterial and antioxidant activities and can be used as alternative antibiotics or potential feed additives for the control of animal pathogenic bacteria.

Development of non-destructive measurement method for discriminating disease-infected seed potato using visible/near-Infrared reflectance technique (광 반사방식을 이용한 감염 씨감자 비파괴 선별 기술 개발)

  • Kim, Dae-Yong;Cho, Byoung-Kwan;Lee, Youn-Su
    • Korean Journal of Agricultural Science
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    • v.39 no.1
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    • pp.117-123
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    • 2012
  • Pathogenic fungi and bacteria such as Pectobacterium atrosepticum, Clavibacter michiganensis subsp. sepedonicus, Verticillium albo-atrum, and Rhizoctonia solani were the major microorganism which causes diseases in seed potato during postharvest process. Current detection method for disease-infected seed potato relies on human inspection, which is subjective, inaccurate and labor-intensive method. In this study, a reflectance spectroscopy was used to classify sound and disease-infected seed potatoes with the spectral range from 400 to 1100 nm. Partial least square discriminant analysis (PLS-DA) with various preprocessing methods was used to investigate the feasibility of classification between sound and disease-infected seed potatoes. The classification accuracy was above 97 % for discriminating disease seed potatoes from sound ones. The results show that Vis/NIR reflectance method has good potential for non-destructive sorting for disease-infected seed potatoes.

Molecular Differentiation of Bacillus spp. Antagonistic Against Phytopathogenic Fungi Causing Damping-off Disease

  • Cho, Min-Jeong;Kim, Young-Kwon;Ka, Jong-Ok
    • Journal of Microbiology and Biotechnology
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    • v.14 no.3
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    • pp.599-606
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    • 2004
  • Gram-positive antagonistic bacilli were isolated from agricultural soils for possible use in biocontrol of plant pathogenic fungi, Fusarium oxysporum, Rhizoctonia solani, and/or Pythium ultimum. Among the 65 antagonistic Gram-positive soil isolates, 22 strains were identified as Bacillus species by 16S rDNA sequence analyses. Four strains, including DF14, especially exhibited multiple antagonistic properties against the three damping-off fungi. Genotypic properties of the Bacillus isolates were characterized by rapid molecular fingerprinting methods using repetitive extragenic palindromic-PCR (REP-PCR), ribosomal intergenic spacer-length polymorphisms (RIS-LP), 16S rDNA PCR-restriction fragment length polymorphisms (PCR-RFLP), and strain-specific PCR assays. The results indicated that the REP-PCR method was more valuable than the RIS-LP and 16S rDNA PCR-RFLP analyses as a rapid and reliable approach for bacilli typing and identification. The use of strain-specific primers designed based on 16S rDNA sequence comparisons enabled it to be possible to selectively detect a strain, DF14, which is being used as a biocontrol agent against damping-off fungi.

Standard Methods for the Detection and Assessment of Safety in Milk and Dairy Products in Korea (우유 및 유제품의 안전성 평가를 위한 병원성미생물 검사법)

  • Kim, Hyoun-Wook;Seol, Kuk-Hwan;Ham, Jun-Sang;Jang, Ae-Ra;Kim, Dong-Hun;Oh, Mi-Hwa
    • Journal of Dairy Science and Biotechnology
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    • v.29 no.2
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    • pp.59-68
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    • 2011
  • In Korea, there are a couple of risk assessment organizations: The Animal Plant and Fisheries Quarantine and Inspection Agency(QIA) and the Korea Food and Drug Administration (KFDA). The major food laws include the Food Sanitation Act (FSA), the Livestock Product Processing Act (LPPA), and the Agricultural Products Quality Control Act (APQCA). Milk and dairy products are mostly controlled by the Food Sanitation Act and the Livestock Product Processing Act. This study was carried out to estimate the current standard methods of foodborne pathogens for dairy products, comparing the Livestock Products Processing Act with the Food Sanitation Act. The standard methods of foodborne pathogens for dairy products are composed by growth culture, isolation culture, and identification, however, standard methods of QIA and KFDA are different at the using of medium and inspection stage. Therefore, consolidation of risk management and risk assessment methods are regarded important to provide safe dairy products to consumer.

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Study on Pharmaceutical Byproducts and Sludges for Practical Application as Raw materials of organic compost (제약업종 부산물 및 오니의 퇴비원료로 활용 가능성 연구)

  • Lim, Dong-Kyu;Lee, Sang-Beom;Nam, Jae-Jag;Na, Young-Eun;Kwon, Jang-Sik;Cho, Nam-Sung;Park, Myoung-Han
    • Journal of the Korea Organic Resources Recycling Association
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    • v.11 no.2
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    • pp.86-96
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    • 2003
  • This study was conducted to investigate use possibility of pharmaceutical byproducts(process sludge) and waste water sludges as raw materials of organic compost at Fertilizer Official Regulation of Fertilizer Management Law in 2002. All pharmaceutical byproducts were satisfied the standard levels of raw material regulated in organic compost, some waste water sludges were deficient in the level. The content of n-hexane extractable material(HEM) was in byproduct higher than in waste water sludge. This was presumed that the sludge with containing a lot of organic matter was high in HEM content. Of the whole microorganism flora, bacteria was mainly detected, and yeast and filamentous fungi took up less population which was fluctuated depending on the source of sludges. Most dominated bacteria were identified into Genus of Pseudomonas. Pseudomonas syringae and Rathayibacter bathayi was classified as plant pathogenic bacteria.

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In vitro Evaluation of the Mechanism of Antagonism and Phosphate Solubilization by the Insect Gut Bacteria Pseudomonas sp. PRGB06 that Exhibits Plant Growth Promotion and Bio-Fertilizing Traits (배추좀나방 내장에서 분리한 식물생장촉진미생물 Pseudomonas sp. PRGB06의 길항기작과 인산가용화의 기내 평가)

  • Kim, Kyoung-A;Indiragandhi, P.;Anandham, R.;Palaniappan, P.;Trivedi, P.;Madhaiyan, M.;Han, Gwang-Hyun;Sa, Tong-Min
    • Korean Journal of Soil Science and Fertilizer
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    • v.41 no.1
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    • pp.18-25
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    • 2008
  • Pseudomonas sp. PRGB06, a bacterial strain isolated from diamondback moth (Plutella xylostella) gut, was examined for its plant growth promotion and biofertilizing traits. The bacteria growth was observed under various conditions of carbon sources, temperature, pH and salt concentrations. In addition, the mechanisms of antagonism and phosphate solubilization were investigated. The bacterial strain PRGB06, grew well using most of the tested carbon sources. The best growth was observed at $30^{\circ}C$ and pH 7. The inhibition of the pathogenic fungi was likely due to the volatile antifungal metabolite and ammonia gas produced by the bacteria. A significant positive relationship was found between the phosphate solubilization and acid production. When inoculated with PRGB06 in vitro and in gnotobiotic condition, red pepper and maize showed increase in root length, seedling vigor and dry bio-mass.

Microbial Contamination in a Fresh-Cut Onion Processing Facility (신선편이 양파 가공작업장 내의 시설 및 공정별 미생물 오염 실태)

  • Lee, Hye-Ok;Kim, Ji-Young;Yoon, Doo-Hyun;Cha, Hwan-Soo;Kim, Gun-Hee;Kim, Byeong-Sam
    • Food Science and Preservation
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    • v.16 no.4
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    • pp.567-572
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    • 2009
  • We evaluated the microbiological quality of a facility in which freshly cut onions were prepared. The total plate counts on walls, equipment, and raw materials were ND (not detected) to $10^1$ CFU/100 $cm^2$, $10^0{\sim}10^3$ CFU/100 $cm^2$, and $10^3{\sim}10^4$ CFU/g, respectively. No coliforms were detected on walls however, coliforms were detected at concentrations of ND to $10^3$ CFU/100 $cm^2$ and $10^3{\sim}10^4$ CFU/g on equipment and raw materials, respectively. The total plate counts for falling and floating bacteria in the processing plant were ND to $10^0$CFU/plate and $10^1{\sim}10^2$ $CFU/m^3$, respectively. Pathogenic microorganisms such as Escherichia coli, Salmonella spp, Staphylococcus aureus, and Listeria monocytogenes were not detected on walls, equipment, or raw materials. Overall, the results of the study indicate that hygiene control at the fresh-cut processing plant should be improved.

A Putative Early Response of Antifungal Bacillus lentimorbus WJ5 Against the Plant Pathogenic Fungus, Colletotrichum gloeosporioides, Analyzed by a DNA Microarray

  • Lee Young-Keun;Jang Yu-Sin;Chang Hwa-Hyoung;Hyung Seok Won;Chung Hye-Young
    • Journal of Microbiology
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    • v.43 no.3
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    • pp.308-312
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    • 2005
  • The global RNA transcription profiles of Bacillus lentimorbus WJ5 under an in vitro co-culture with Colletotrichum gloeosporioides were analyzed in order to study the antagonistic bacteria-fungi interactions. Using a filter membrane system, B. lentimorhus WJ5 was exposed to the spores of C. gloeosporioides at the late exponential stage. The transcription profiles of the B. lentimorhus WJ5, both with and without a challenge from C. gloeosporioides, were analyzed using custom DNA chips containing 2,000 genome fragments. A total of 337 genes were expressed, with 87 and 47 up- and down-regulated, respectively. Of these, 12 genes, which were involved in central carbon metabolisms, and 7 from minor catabolism were relatively highly up-regulated (> 10 fold) and down-regulated (< 0.2 fold), respectively. Nine genes, which were thought to be related to the antifungal activity, were also up-regulated, but their levels were not so high (2.0 - 9.7 folds). From the results, during the early stage of the co-culture of B. lentimorbus WJ5 and C. gloeosporioides, nutrient competition seemed to occur; therefore, the genes from central carbon metabolisms could be up-regulated, while those from minor catabolism could be down-regulated.

Antioxidant and Antibacterial Effects of Carnosic Acid on White Sausage (카르노스산에 의한 화이트소시지의 항산화 및 항균 효과)

  • Lee, Jo-Won;Choe, Il-Shin;Kim, Woan-Sub
    • Korean Journal of Organic Agriculture
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    • v.21 no.2
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    • pp.219-232
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    • 2013
  • The relevant main constituents of rosemary are compound of a vast number of polyphenolics, including carnosic acid, carnosol, rosmarinic acid, and ursolic acid. Recently, phenolic compounds of plant origin have attracted considerable attention due to their beneficial functional and nutritional effects including antioxidant and antibacterial activity. This study was carried out to investigated effect of carnosic acid on pH, thiobarbituric acid reactive substance (TBARS), volatile basic nitrogen (VBN), 2,2-diphenyl-1-picryhydrazyl (DPPH) radical scavenging activity, and total bacteria counts in white sausage during the storage at room temperature. Addition of carnosic acid was decreased of pH, TBARS, VBN, and enhanced antioxidant and antibacterial activity in white sausage. These activities increased with increasing concentrations of carnosic acid. Furthermore, the potent antibacterial activities of carnosic acid against pathogenic bacteria (Escherichia coli KCCM 11234 and Salmonella enteritidis KCCM 12021) were measured. Carnosic acid showed noticeable inhibitory effects on E. coli and S. enteritidis. In conclusion, carnosic acid might be used as a natural preservative in white sausage.