• The Korean Journal of Mycology
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• v.36 no.2
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• pp.138-143
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• 2008

Botrytis cinerea, gray mold pathogen, causes serious losses in greenhouse tomato crop. In this study, a primer set was developed for identification and specific PCR detection of B. cinerea from tomato plants. The primer pair (BTF1/BTR1) was designed from polymorphic sequence region in pyruvate carboxylase gene (pyc) of B. cinerea. A PCR product (112 bp) was amplified on genomic DNA of 13 B. cinerea isolates from 10 different host plants, but not on those from 6 other Botrytis spp., 4 Botryotinia spp., 5 Sclerotinia spp. and 16 other genus of phytopathogenic fungi. The sensitivity limit of the primer set was 2 pg of genomic DNA of B. cinerea, approximately. The PCR assay using species-specific primer set was specifically able to detect the pathogen on naturally infected tomato plants and artificially infected plants. These results suggest that the sensitivity and specificity of this primer set can be applied in a rapid and accurate diagnosis of tomato disease caused by B. cinerea.

• KOREAN POULTRY JOURNAL
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• v.34 no.4 s.390
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• pp.104-106
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• 2002

미국이 자국내 철강제 수입에 대한 관세부과 조치가 세계적인 문제가 되고 있는 시점에서, 러시아에 수출되고 있는 미국산 닭고기에서 살모넬라 등 위생에 문제가 될 수 있는 병원균이 검출되었다는 보도를 미국은 무역보복조치로 해석하고 있어 무역전쟁을 실감케 하고 있다. 축산물에 대한 안전이 식품의 안전성에 직결된다는 의식을 갖고 있는 우리나라의 배합사료제조회사들은, 자발적으로 가축에게 급여하는 사료에 대한 안전과 위생관리를 위하여 ISO는 물론 까다로운 것으로 알려진 HACCP 제도를 도입하고 있다. 이러한 노력이 종합적이고 기본적인 단계에 해당하는 양계농가에서도 함께 진행될 때 더욱 효과적인 결과를 얻을 수 있으리라 생각되어 살모넬라에 대한 일본의 농가단위 위생대책을 소개한다.

• Journal of the Korean Chemical Society
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• v.55 no.2
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• pp.262-267
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• 2011

The denaturing high performance liquid chromatography(DHPLC) with fluorescence detector assay is very useful tool for detecting nucleic acids. Furthermore, loop-mediated isothermal amplification(LAMP) constitutes a potentially valuable tool for rapid diagnosis of pathogenic microorganisms. In this study, we evaluated the specificity, detection limit, and sensitivity of a LAMP method and DHPLC method for rapid detection of the hepatitis b virus(HBV). As a result, the LAMP assay reported here has the advantage of rapid detection whereas, DHPLC assay has more sensitivity than other assays. These findings suggest that LAMP and DHPLC assay may be good tool for rapid diagnosis of clinical HBV infection.

• The Korean Journal of Mycology
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• v.46 no.4
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• pp.467-477
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• 2018

Colletotrichum circinans, an anthracnose pathogen, causes serious damage to onions worldwide. In this study, specific molecular markers were developed to detect C. circinans accurately and quickly with both conventional and real-time PCR methods. The cirTef-F/cirTef-R and cirTu-F/cirTu-R primer sets, which are specific for C. circinans, were constructed by analyzing $tef-1{\alpha}$ and ${\beta}-tubulin$ genes in the fungus. Using the conventional PCR method, 100 pg and 1 ng of fungal DNA could be detected using the cirTef-F/cirTef-R and cirTu-F/cirTu-R sets, respectively. Using the real-time PCR method, 10 pg and 100 pg of fungal DNA could be detected more sensitively with the cirTef-F/cirTef-R and cirTu-F/cirTu-R sets, respectively. Detection of C. circinans from the artificially infected onion seeds was possible by using both conventional and real-time PCR methods and the developed cirTef-F/cirTef-R primer set. The PCR markers specific for C. circinans developed in this study may enhance the efficiency of fungal pathogen detection in imported vegetables and seeds.

• Journal of Food Hygiene and Safety
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• v.16 no.4
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• pp.251-257
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• 2001

Listeria monocytogenes and L. ivanovii are important food-pathogens for human and animal. The diagnostic of Listeria in food using culture medium requires time and laborwork, because there are many other non-pathogenic species like L. innocua, L welshimeri, L. seeligeri and L. grayi in Genus Listeria. For these reasons, Lismix multiplex PCR method was developed as a rapid method for the detection and identification of Listeria. In this study we developed a practical system of Lis-mix PCR detection for the application to food samples and new developed Siw-mix III PCR system overall 69 listerial strains were successful species-identified and confirmed. Also, the Siw-mix III PCR system allows the species-specific identification among L. ivanovii, L. welshimeri and L seeligeri in a single PCR.

• Korean Journal of Agricultural Science
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• v.9 no.1
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• pp.275-283
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• 1982

One hundred and sixteen seed samples of 16 different crops were tested by routine seed health testing methods. Seeds of rice, soybean and barley were found to be associated with 18, 15 and 7 species of fungi, respectively. Discoloration and spotting of various kinds were observed in rice seeds. Epicoccum purpurascens, Pyricularia oryzae and Trichoconiella pedwickii were higher in these discoloured than in normal seeds of rice. Macrophomina phaseolina, the charcoal-rot fungus is a new record for soybean in Korea. Seeds of sesame were infected by 13 fungi Alternaria sesami, A. sesarmicola and Corynespora cassiicola were detected predominantly. Twenty one species of fungi were detected in seed samples of 12 commercial vegetable crops. Alternaria, Colletotrichum and Fusarium were found predominantly. Some seed samples of commercial vegetable crops were heavily infected with pathogenic fungi and showed heavy seed and seedling rot.

• The Korean Journal of Pesticide Science
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• v.19 no.2
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• pp.119-124
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• 2015

Bacterial leaf spot, caused by Pseudomonas syringae pv. syrinage, is a very damaging disease to green pumpkin in Gong-ju and Non-san nursery. However, there is no good method to control the disease in Korea. Growth inhibition of pathogen on medium, control efficacy on seedling stage, and seed treatment effect of 6 anti-bacterial pesticides were investigated for selection of the best pesticide for seed treatment and control of the disease. Growth inhibition zone on King's B medium were the largest by oxytetracycline 170 ppm and oxytetracycline 15 ppm + streptomycin sulfate 188 ppm, oxolinic acid 200 ppm, streptomycin 200 ppm were next respectively. Control efficacy of oxytetracycline 1.5% + streptomycin sulfate 18.8% WP and oxytetracycline 17% WP on seedling stage were 71.4% and 49.4%, respectively. Seed treatment of oxytetracycline 15 ppm + streptomycin sulfate 188 ppm on the artificially inoculated seeds inhibits pathogen growth completely from the treated seeds and 96% control efficacy on grow-out test of the treated seeds. Seed treatment of streptomycin 100 ppm (2,000 dilution of streptomycin 20%) on the artificially inoculated seeds allow 280 cfu/g of pathogen growth from the treated seeds and 60% control efficacy on grow-out test of the treated seeds. Seed treatment of oxytetracycline 85 ppm (2000 dilution of oxytetracycline 17% WP) on the artificially inoculated seeds allow 80 cfu/g of pathogen growth from the treated seeds and 90% control efficacy on grow-out test of the treated seeds. These results suggested that oxytetracycline 1.5% + streptomycin sulfate 18.8% WP was the best pesticide for seed treatment to control of the bacterial spot disease by Pseudomonas syringae pv. syringae.

• Research in Plant Disease
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• v.17 no.2
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• pp.184-190
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• 2011

Ralstonia solanacearum, a causal agent of bacterium wilt is very difficult to control once the disease becomes endemic. Thus, Ralstonia solanacearum is a plant quarantine bacterium in many countries including Korea. In this study, we developed PCR assays, which can detect Ralstonia solanacearum from the Solanaceae seeds. Primers RS-JH-F and RS-JH-R amplified specifically a 401 bp fragment only from Ralstonia solanacearum race 1 and race 3. The nested PCR primers, RS-JH-F-ne and RS-JH-R-ne that were designed inside of 1st PCR amplicon amplified specifically a 131 bp fragment only from Ralstonia solanacearum race 1 and race 3. The primers did not amplify any non-specific DNA from the seed extracts of the Solanaceae including tomato and pepper. When detection sensitivity were compared using the Solanaceae seeds inoculated with target bacteria artificially, the nested PCR method developed in this study 100 times more sensitive than ELISA and selective medium. Therefore, we believe that the PCR assays developed in this work is very useful to detect Ralstonia solanacearum in the Solanaceae seeds.

• Research in Plant Disease
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• v.16 no.2
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• pp.129-135
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• 2010

PCR primers were developed to detect Pseudomonas savastanoi pv. phaseolicola, a causal agent of halo blight that occurs in all species of common bean (Phaseolus vulgaris L.), from the bean seeds. A primer set, Psp-JHF and Psp-JH-R, specifically amplified 513 bp fragment from Pseudomonas savastanoi pv. phaseolicola only. A nested primer set, psp-JH-F-ne and psp-JH-R-ne, designed from the $1^{st}$ PCR amplicon, amplified 169 bp fragment. The primer sets did not amplify any non-specific DNA from the seed extracts of Fabaceae including 4 beans, 2 soybeans, and 2 peas. The detection sensitivity of the nested PCR method developed in this study was much higher than that of ELISA and selective medium. PCR assays developed in this study should be useful to detect Pseudomonas savastanoi pv. phasolicola from the bean seeds.

• The Korean Journal of Pesticide Science
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• v.12 no.2
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• pp.168-176
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• 2008

Recently damage of rice bakanae disease disseminated by infected seeds increased in paddy field in Korea. For controlling rice bakanae disease, the efficacy of 17 fungicides was assessed by 5 kinds of bioassay, spore germination test (SGT), mycelial growth test, detection test on Komada's medium (KDT), pouch test (PT) and greenhouse test (GT). Among ergosterol biosynthesis inhibiting fungicides, prochloraz showed a high controlling activity in all the assay systems while the others showed very low activity except for $500\;{\mu}g/ml$ of hexaconazole in GT and $500\;{\mu}g/ml$ of triflumizole in KDT. Although benomyl and the mixture of benomyl and thiram showed a good activity at 100 and $500\;{\mu}g/ml$ in SGT and PT, respectively, in GT they did a middle activity. Trifloxystrobin and kresoxim-methyl included in strobilurins showed a good activity even at $20\;{\mu}g/ml$ in KDT as well as a middle activity in SGT. Also a high activity not only at $10\;{\mu}g/ml$ in SGT but also at $100\;{\mu}g/ml$ KDT was detected in thiram. The activity of fludioxonil was confirmed in SGT, KDT and PT. Based on these results, it is very important to determine a bioassay system, because the fungicidal activity against rice bakanae disease was fluctuated depending on a assay systems as well as the mechanism of fungicide.