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Suitable Conditions for Mycelial Culture of Tremella fuciformis (흰목이 균사체 배양 적합 조건 설정)

  • Lee, Eun-ji;Park, Hye-Sung;Lee, Chan-Jung;Kong, Won-Sik;Koo, Chang-Duck
    • The Korean Journal of Mycology
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    • v.47 no.1
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    • pp.1-12
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    • 2019
  • This study was conducted to investigate the optimum culture conditions, for mass production of Tremella fuciformis in M9 basic medium. The strain KMCC04674, used in this study was identified T. fuciformis by internal transcribed spacer (ITS). To define the optimum conditions for the mass production of T. fuciformis, we investigated the effects of different culture conditions and various nutrient sources on the fungal growth. The optimum initial pH and temperature for the fungal growth were 5.0 and $25^{\circ}C$, respectively. The optimal composition of the growth medium was 4.0% mannitol, 3.0% $NH_4H_2PO_4$, 1.0% malt extract, 1.0% glutamic acid, 5mM $CaCl_2$, and 0.5% glucanic acid.

Isolation of Wild Yeasts from Riversides in Geumgang Upstream, Taean and Suncheonman Seashores and Microbiological Characteristics of the Unrecorded Wild Yeasts (금강 상류와 태안, 순천만 해변들로부터 야생효모의 분리 및 국내 미기록 효모들의 균학적 특성)

  • Park, Seon-Jeong;Jang, Ji-Eun;Lee, Hyang-Burm;Lee, Jong-Soo
    • The Korean Journal of Mycology
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    • v.49 no.1
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    • pp.67-79
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    • 2021
  • This study aimed to isolate wild yeasts from waters and soils on the riversides of Geumgang upstream in Geumsan, seashores of Cheongpodae in Taean and Suncheonman in Suncheon, Korea, and to investigate their unrecorded microbiological characteristics. Forty-nine species of wild yeasts, including Aureobasidium pullans YH 4-3, were isolated from 40 samples of Cheonnaegang in Geumgang upstream. Fifty-six species and 36 species of wild yeasts were also isolated from 80 samples of Cheongpodae Beach and Suncheonman Mashland, respectively. Species from Candida genus were isolated from all three locations. Among them, Candida michaelii NNIBRFG28278, Sporobolomyces japonicus NNIBRFG28271, Dioszegia buhagiarii NNIBRFG28279, Ustilago spermophora NNIBRFG28273, Nakazawaea pomicola NNIBRFG31590, Candida natalensis NNIBRFG31591, Candida pseudorhagii NNIBRFG31592, Candida santamariae NNIBRFG31593, Cutaneotrichosporon terricola NNIBRFG31594, and Meira nashicola NNIBRFG31595 represent newly recorded yeast strains in Korea. Almost all of these unrecorded yeasts were oval shaped, and Sporobolomyces japonicus NNIBRFG28271 and Candida natalensis NNIBRFG31591 have ascospores. All the strains grew well in yeast extract-peptone-dextrose (YPD) and yeast extract-malt extract (YM) media, and Ustilago spermophora NNIBRFG28273 grew well in vitamin-free medium. Sporobolomyces japonicus NNIBRFG28271 grew well in 15% NaCl-containing YPD medium, and Ustilago spermophora NNIBRFG28273 and Dioszegia buhagiarii NNIBRFG28279 assimilated starch, and Ustilago spermophora NNIBRFG28273 produced ethanol.

Medium optimization for growth of Bacillus amyloliquefaciens ISP-5 strain and evaluation of plant growth promotion using lettuce (Bacillus amyloliquefaciens ISP-5 균주의 배지 최적화 및 상추를 이용한 식물 생장 촉진 평가)

  • Kang-Hyun Choi;Sun Il Seo;Haeseong Park;Ji-hwan Lim;Pyoung Il Kim
    • Journal of Plant Biotechnology
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    • v.49 no.4
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    • pp.356-361
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    • 2022
  • Bacillus sp. is a useful strain for agriculture because it promotes plant growth and controls plant pathogens through a variety of mechanisms. In this study, we obtained a microbial preparation with a high number of viable cells by culturing newly isolated soil bacteria on an optimized medium. Subsequently, we applied this preparation to lettuce to enhance its growth and yield. First, B. amyloliquefaciens ISP-5 was isolated from soil. Next, optimization of culture medium was carried out using 5 L scale fermenters. When culturing B. amyloliquefaciens ISP-5 on this optimized medium, the number of viable cells was approximately 1000 times higher than that obtained from culturing on the commercial medium. Afterwards, the plant growth promotion properties of the ISP-5 strain were evaluated using lettuce as a test plant. Foliar spray treatment of lettuce was carried out by inoculating half the standard concentration suspension (0.5 × 107 cfu/ml). As a result, leaf width increased by 8.6% and leaf length increased by 12.9% compared to the control group. Live weight also increased by 24.2% and dry weight by 23.9%. Considering the results from field test, B. amyloliquefaciens ISP-5 showed potential as a plant growth-promoting bacteria.

Six Yeast Strains Isolated from Freshwaters for the First Record in Korea (국내 담수환경에서 분리된 국내 미기록 효모 6종 보고)

  • Yoosun Oh;Min Jeong Seo;Jaeduk Goh;Hye Yeon Mun
    • The Korean Journal of Mycology
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    • v.50 no.3
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    • pp.217-224
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    • 2022
  • This study aimed to isolated wild yeasts from freshwater in Korea. The strains were identified by using the D1/D2 domains of the 26S rDNA regions. Consequently, six strains were named as Apiotrichum chiropterorum (NNIBRFG36995), A. domesticum (NNIBRFG32938), A. dulcitum (NNIBRFG33144), A. laibachii (NNIBRFG36991), Saprochaete quercus (NNIBRFG33183) and Tausonia pullulans (NNIBRFG33181). These yeasts have not previously been recorded in Korea, this paper is the first report. We were investigated to the morphological and cultural characteristics of these yeasts. All the strains grew well on Yeast extract peptone dextrose (YPD), Potato dextrose (PD), and Yeast mold (YM) media and in temperature range of 15-30℃. A. domesticum (NNIBRFG32938), A. laibachii (NNIBRFG36991) and T. pullulans (NNIBRFG33181) grew even in 20% glucose containing YPD medium, so they had glucose tolerance. A. domesticum (NNIBRFG32938) and A. laibachii (NNIBRFG36991) had salt resistance as growing even in 5% NaCl containing YPD medium.

Isolating and characterizing the unrecorded Wild Yeasts from Seawater and Soil in Haeundae and Mongdol Beaches on the Southern Coast of, Korea (남해안 해운대와 몽돌 해수욕장 주변환경으로부터 야생 효모의 분리 및 국내 미기록 효모들의 균학적 특성)

  • Seon-Jeong Park;Ji-Eun Jang;Jeong-Su Moon;Hyang-Burm Lee;Jong-Soo Lee
    • The Korean Journal of Mycology
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    • v.50 no.1
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    • pp.65-73
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    • 2022
  • This study aimed to isolate wild yeasts from seawaters and soils samples of the Haeundae and Mongdol beaches on the southern coast of Korea, and to characterize these unrecorded wild yeast strains. In total, 41 strains, representing 37 different species of wild yeast were isolated from 70 samples collected from the beaches. Among these, 14 strains were isolated from the alkalophilic medium of yeast extract-peptone-dextrose (YPD) medium (pH 9.0), and 27 strains were isolated concurrently on general YPD medium (pH 6.5). Among the 41 isolated wild yeast strains, Candida insectorum HUD 16-3(JSL-KSS-002) and Metschnikowia citriensis HUD 12-5(JSL-KSS-001) had not previously been recorded. We investigated the microbiological characteristics of these two unrecorded yeast strains and three other strains-, Cystobasidium lysinophilum JSC 52-2(JSL-GGU-019), Candida takata NMD 11-1(JSL-GGU-017) and Candida panamensis ASG 58M-2(JSL-GGU-018) from Jangseoncheon in Jellabuk-do and Jangtaesan in Deajeon city. All five previously unrecorded yeasts were oval and did not form spores. All strains grew well in YPD and yeast extract-malt extract media in a vitamin-free medium. Two strains, including C. insectorum HUD 16-3(JSL-KSS-002) grew well in a 15% NaCl-containing YPD medium. Three strains, including Cys. lysinophilum JSC52-2(JSL-GGU-019) assimilated lactose, and all strains assimilated starch.

Microbiological Characteristics, Enzyme Activity, and Physiological Functionality of Unrecorded Wild Yeasts from the Wolsung Valley of South Deogyu Mountain and Jinjamcheon in Daejeon, Korea (남덕유산 월성계곡과 대전광역시 진잠천주변 등으로부터 선발된 국내 미기록 야생효모들의 균학적 특성과 주요 효소활성 및 생리활성)

  • Jeong-Su Moon;Min-Young An;Mal-Geum Park;Jong-Soo Lee
    • The Korean Journal of Mycology
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    • v.51 no.1
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    • pp.39-49
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    • 2023
  • This study aimed to screen and investigate the microbiological characteristics, enzyme activity, and physiological functionality of unrecorded wild yeasts from the Wolsung valley of South Deogyu mountain and Jinjamcheon in Daejeon city, Korea. Candida sorboxylosa WSC25-4 (NNIBRFG47336), Arxula adeninivorans WSC6-3 (NNIBRFG47335), Farysizyma taiwaniana JRC6-2 (NNIBRFG47339), Saturnispora dispora JTS19-1 (NNIBRFG47334), Vanderwaltozyma polyspora JJCH5-3 (NNIBRFG47337), and Vanrija fragicola SW-9 (NNIBRFG47338) have not been previously recorded. None of these formed spores or pseudo-mycelium, and four strains, including C. sorboxylosa WSC25-4, were global in shaped. Except for V. polyspora JJCH5-3, all strains grew well in the yeast extract-peptone-dextrose (YPD), potato-dextrose and yeast malt extract media. A. adeninivorans WSC6-3 and S. dispora JTS19-1 grew well in 40% glucose-containing YPD medium and V. fragicola SW-9 in 5% NaCl-containing YPD medium. The supernatant of A. adeninivorans WSC6-3, F. taiwaniana JRC6-2, and V. fragicola SW-9 exhibited 58.0, 59.0, and 59.0% of glucoamylase activity, respectively. C. sorboxylosa WSC25-4 also produced extracellular alkaline protease. Antioxidant activity of the supernatant from F. taiwanian JRC6-2 was high (79.8%), while the other five unrecorded yeasts demonstrated elevated antiaging superoxide dismutase (SOD)-like activity (99%). Anti-gout xanthin oxidase inhibitory activity of S. dispora JTS19-1 and V. fragicola SW-9 was also elevated at 79 and 80%, respectively.

Physiological Functionalities and Enzyme Activities of Non-pathogenic Pigmented Wild Yeasts (비병원성 야생 색소 효모들의 생리활성과 효소활성)

  • Jeong-Su Moon;Seung-A Shin;Seung-Eun Baek;Jong-Soo Lee
    • The Korean Journal of Mycology
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    • v.51 no.3
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    • pp.205-217
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    • 2023
  • This study aimed to produce novel bioactive compounds from non-pathogenic pigmented wild yeasts. Culture supernatants and cell-free extracts of non-pathogenic pigmented yeast strains were prepared, and their physiological functionalities and enzyme activities were measured. Cell-free extracts from Rhodosporidium paludigenum HHGG35-1 and culture supernatants from Rhodosporidium diobovatum NMD18-1 demonstrated very high antioxidant activity (76.6%) and anti-gout xanthin oxidase inhibitory activity (86.2%), respectively. Maximal production of the antioxidants (76.9%) was obtained when Rh. Paludigenum HHGG35-1 was cultured in a yeasts extract-peptone-dextrose (YPD) medium (pH 6.5) at 30℃ for 24 h. The xanthin oxidase inhibitor was also maximally produced (91.6%) when Rh. Diobovatum NMD18-1 was cultured at 30℃ for 96h in a YPD medium (pH 6.5). Rh. Paludigenum HHGG35-1 was oval in shape and formed ascospre. The Rh.diobovatum NMD18-1 specimen displayed dimensions of 1.6 × 1.6 ㎛ and produced ascospores; however, it did not form pseudomycelium. Both of Rh. Paludigenum HHGG35-1 and Rh. Diobovatum NMD18-1 grew well in a 40%-glucose-containing YPD medium and 10%-NaCl-containing YPD medium.

Studies on the physio-chemical properties and the cultivation of oyster mushroom(Pleurotus ostreatus) (느타리버섯의 생리화학적성질(生理化學的性質) 및 재배(栽培)에 관(關)한 연구(硏究))

  • Hong, Jai-Sik
    • Applied Biological Chemistry
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    • v.21 no.3
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    • pp.150-184
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    • 1978
  • Nutritional characteristics and physio-chemical properties of mycelial growth and fruitbody formation of oyster mushroom(Pleurotus ostreatus)in synthetic media, the curtural condition for the commerical production in the rice straw and poplar sawdust media, and the changes of the chemical components of the media and mushroom during the cultivation were investigated. The results can be summarized as follows: 1. Among the carbon sources mannitol and sucrose gave rapid mycelial growth and rapid formation of fruit-body with higher yield, while lactose and rhamnose gave no mycelial growth. Also, citric acid, succinic acid, ethyl alcohol and glycerol gave poor fruit-body formation, and acetic acid, formic acid, fumaric acid, n-butyl alcohol, n-propyl alcohol and iso-butyl alcohol inhibited mycelial growth. 2. Among the nitrogen sources peptone gave rapid mycelial growth and rapid formation of fruit-body with higher yield, while D,L-alanine, asparatic acid, glycine and serine gave very poor fruit-body formation, and nitrite nitrogens, L-tryptophan and L-tyrosine inhibited mycelial growth. Inorganic nitrogens and amino acids added to peptone were effective for fruit-body growth, and thus addition of ammonium sulfate, ammonium tartarate, D,L-alanine and L-leucine resulted in about 10% increase fruit-body yield. L-asparic acid about 15%, L-arginine about 20%, L-glutamic acid, and L-lysine about 25%. 3. At C/N ratio of 15.23 fruit-body formation was fast, but the yield decreased, and at C/N ratio of 11.42 fruit-body formation was slow, but the yield increased. Also, at the same C/N ratio the higher the concentration of mannitol and petone, the higher yield was produced. Thus, from the view point of both yield of fruit-body and time required for fruiting the optimum C/N ratio would be 30. 46. 4. Thiamine, potassium dihydrogen phosphate and magnecium sulfate at the concentration of $50{\mu}g%$. 0.2% and 0.02-0.03%, respectively, gave excellent mycelial and fruit-body growth. Among the micronutrients ferrous sulfate, zinc sulfate and manganese sulfate showed synergetic growth promoting effect but lack of manganese resulted in a little reduction in mycelial and fruit-body growth. The optimum concentrati on of each these nutrients was 0.02mg%. 5. Cytosine and indole acetic acid at 0.2-1mg% and 0.01mg%, respectively, increased amount of mycelia, but had no effect on yield of fruit-body. The other purine and pyrimidine bases and plant hormones also had no effect on mycelial and fruit-belly yield. 6. Illumination inhibited mycelial growth, but illumination during the latter part of vegetative growth induced primordia formation. The optimum light intensity and exposure time was 100 to 500 lux and 6-12 hours per day, respectively. Higher intensity of light was injurous, and in darkness only vegetative growth without primordia formation was continued. 7. The optimum temperature for mycelial growth was $25^{\circ}C$ and for fruit-body formation 10 to $15^{\circi}C$. The optimum pH range was from 5.0 to 6.5. The most excellent fry it-body formation were produced from the mycelium grown for 7 to 10 days. The lesser the volume of media, the more rapid the formation of fruit-body; and the lower the yield of fruit-body; and the more the volume of media, the slower the formation of fruit-body, and the higher the yield of fruit-body. The primordia formation was inhibited by $CO_2$. 8. The optimum moisture content for mycelial growth was over 70% in the bottle media of rice straw and poplar sawdust. 10% addition of rice bran to the media exhibited excellent mycelial growth and fruit-body formation, and the addition of calciumcarbonate alone was effective, but the addition of calcium carbonate was ineffective in the presence of rice bran. 9. In the cultivation experiments the total yield of mushroom from the rice straw media was $14.99kg/m^2$, and from the sawdust media $6.52kg/m^2$, 90% of which was produced from the first and second cropping period. The total yield from the rice straw media was about 2.3 times as high as that from the sawdust media. 10. Among the chemical components of the media little change was observed in the content of ash on the dry weight basis, and organic matter content decreased as the cultivation progressed. Moisture content, which was about 79% at the time of spawning, decreased a little during the period of mycelial propagation, after which no change was observed. 11. During the period from spawning to the fourth cropping about 16.7% of the dry matter, about 19.3% of organic matter, and about 40% of nitrogen were lost from the rice straw media; about 7.5% of dry mallet, about 7.6% of organic matter, and about 20% of nitrogen were lost from the sawdust media. For the production of 1kg of mushroom about 232g of organic matter and about 7.0g of nitrogen were consumed from the rice straw media; about 235g of organic matter and about 6.8g of nitrogen were consumed from the sawdust media, 1㎏ of mushroom from either of media contains 82.4 and 82.3g of organic matter and 5.6 and 5.4g of nitrogen, respectively. 12. Total nitrogen content of the two media decreased gradually as the cultivation progressed, and total loss of insoluble nitrogen was greater than that of soluble nitrogen. Content of amino nitrogen continued to increase up to the third cropping time, after which it decreased. 13. In the rice straw media 28.0 and 13.8% of the total pentosan and ${\alpha}$-cellulose, respectively, lost during the whole cultivation period was lost during the period of mycelial growth; in the sawdust media 24.1 and 11.9% of the total pentosan and ${\alpha}$-cellulose, respectively, was lost during the period of mycelial growth. Lignin content in the media began to decrease slightly from the second cropping time, while the content of reduced sugar, trehalose and mannitol continued to increase. C/N ratio of the rice straw media decreased from 33.2 at spawining to 30.0 at ending; that of the sawdust media decreased from 61.3 to 60.0. 14. In both media phosphorus, potassium, manganese and zinc decreased, at magnesium, calcium and copper showed irregular changes, and iron had a tendency to be increased. 15. Enzyme activities are much higher in the rice straw media than in the sawdust media. CMC saccharifying and liquefying activity gradually increased from after mycelial propagation to the second cropping, after which it decreased in both media. Xylanase activity rapidly and greatly increased during the second cropping period rather than the first period. At the start of the third cropping period the activity decreased rapidly in the rice straw media, which was not observed in the sawdust media. Protease activity was highest after mycelial propagation, after which it gradually decreased. The pH of the rice straw media decreased from 6.3 at spawning to 5.0 after fourth cropping; that of the sawdust media decreased from 5.7 to 4.9. 16. The contents of all the components except crude fibre of the mushroom from the rice straw media were higher than those from the sawdust media. Little change was observed in the content of the components of mushroom cropped from the first to the third period, but slight decrease was noticed at the fourth cropping.

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Incubation Conditions Affecting Bacteriocin Production of Lactobacillus plantarum K11 Isolated from Dongchimi (동치미에서 분리된 Lactobacillus plantarum K11의 박테리오신 생산에 영향을 미치는 배양 조건)

  • Lim, Sung-Mee;Lee, Gun-Ja;Park, Sun-Mee;Im, Dong-Soon
    • Journal of Food Hygiene and Safety
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    • v.23 no.2
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    • pp.113-120
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    • 2008
  • The influence of incubation temperature, pH and media components on bacteriocin production by Lactobacillus plantarum K11 were investigated. The highest activity was observed in MRS broth, but no bacteriocin activity was obtained in TSB. The bacteriocin was produced from the exponential growth phase and its activity also reached a maximum in MRS broth, but then dropped after 16 hr because of degradation by extracellular proteolytic enzymes or exhaustion of medium nutrients. The optimal temperature and pH for production of bacteriocin were $37^{\circ}C$ and pH 7.0 in MRS broth, respectively. The addition of 0.5 or 1.0% glucose and $0.5{\sim}1.5%$ lactose to MRS resulted in the increase of the bacteriocin production. With 0.5% NaCl and $K_2HPO_4$, the activities were significantly higher than that of control, respectively. However, increasing nitrogen sources such as beef extract, casein, and tryptone and salts such as $NH_4PO_4$, $MgSO_47H_2O$, and $MnSO_4H_2O$, had detected a negative influence upon the bacteriocin production. Consequently, because the bacteriocin produced by L. plantarum K11 was affected by various incubation conditions, the bacteriocin activity of L. plantarum K11 applied in food as a novel starter will be dependent on environmental factors such as fermentation conditions and food ingredients.

Comparison of the Standard Culture Method and Real-time PCR for the Detection of Vibrio parahaemolyticus in Seafoods and Vegetables (해산식품과 채소에서 Vibrio parahaemolyticus 검출을 위한 배지배양법과 real-time PCR의 비교검증)

  • Chon, Jung-Whan;Hyeon, Ji-Yeon;Hwang, In-Gyun;Kwak, Hyo-Sun;Han, Jeong-A;Chung, Yun-Hee;Song, Kwang-Young;Seo, Kun-Ho
    • Korean Journal of Food Science and Technology
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    • v.42 no.3
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    • pp.355-360
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    • 2010
  • Vibrio parahaemolyticus (V. parahaemolyticus), which is commonly found in raw seafood, causes gastroenteritis in humans. Rapid and effective methods have been developed as culture methods require up to 5-7 days. In this study, real-time PCR was compared with the standard culture method for detecting V. parahaemolyticus in seafood and radish sprout samples. Five hundred grams of the samples were artificially contaminated with V. parahaemolyticus then divided into 20 samples. The samples were incubated in alkaline peptone water and then streaked onto thiosulfate-citrate-bile saltssucrose agar. Biochemical tests for suspicious colonies were performed using the API 20NE strip. In parallel, real-time PCR was performed targeting the toxR gene using the enrichment broth. The real-time PCR was sensitive in discriminating V. parahaemolyticus from other foodborne pathogens. The detection limit of the real-time PCR was $10^3\;CFU/mL$ in phosphate-buffered saline. Although the real-time PCR detected more positive samples (76 out of 180, 42%) than the culture method (66 out of 180, 37%), there was no significant statistical difference (p>0.05) between the two methods. In conclusion, real-time PCR assays could be an alternative to the standard culture method for detecting V. parahaemolyticus in seafood and radish sprouts, which has many advantages in terms of detection time, labor, and sensitivity.