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환기조절 및 약제적기살포에 의한 비닐하우스재배 오이에 발생하는 노균병 방제
김용기(Yong-Ki Kim),류재당(Jae-Dang Ryu),류재기(Jae-Gee Ryu),이상엽(Sang Yeob Lee),심홍식(Hong-Sik Shim) 한국농약과학회 2003 농약과학회지 Vol.7 No.3
Survey on plant diseases occurring on cucumber cultivated in plastic film house of experimental farm in Suwon was conducted. Through the survey, occurrence of damping-off, downey mildew, powdery mildew and Fusarium wilt was observed. Especially downey mildew caused by Pseudoperonospora cubensis was the most severe foliar disease of cucumber. To control the disease effectively, effects of installation of ventilation fan and optimal spray timing of a chemical, dimethomorph+copper oxychloride WP, were investigated. Two ventilation fans installed at the front and at the back of plastic film house reduced air relative humidity by about 6.4% and downey mildew incidence by 55.7%. Downey mildew incidence on cucumber from untreated chemicals plot in plastic film house installed with ventilation fan was on a equal level with that from treated chemicals plot with three times application of dimethomorph+copper oxychloride WP in plastic film house without ventilation fan. Meanwhile in order to select optimal chemical application time, dimethomorth+copper oxychloride WP was treated three times at 7 days-interval from three days before the disease occurred, right after the disease occurred, and two days after the disease occurred, respectively. The result showed that dimethomorth+copper oxychloride WP applied to cucumber leaves and stems from three days prior to, right after, two days after occurrence of downey mildew reduced downey mildew incidence by 72.9, 61.8, and 23.7%, respectively. The above results showed that regulation of environmental factors like air relative humidity and preventive application of chemicals should be considered to establish control strategy to downey mildew.
UHPLC를 이용한 볏짚 사일리지와 동계사료작물의 오크라톡신과 제랄레논 분석법 최적화
함현희,문혜연,이경아,이수형,홍성기,이데레사,류재기,Ham, Hyeonheui,Mun, Hye Yeon,Lee, Kyung Ah,Lee, Soohyung,Hong, Sung Kee,Lee, Theresa,Ryu, Jae-Gee 한국초지조사료학회 2016 한국초지조사료학회지 Vol.36 No.4
조사료에서 오크라톡신 A (OTA) 및 제랄레논(ZEA)의 단성분 분석법을 최적화하기 위하여 곰팡이 독소에 오염되지 않은 볏짚 사일리지, 이탈리안 라이그라스, 청보리, 호밀을 대상으로 각 시료 별로 OTA는 50, 250, $5,000{\mu}g/kg$, ZEA는 300, 1,500, $3,000{\mu}g/kg$의 농도로 스파이킹하고 면역친화성 컬럼을 이용하여 독소를 정제한 후 UHPLC-FLD를 이용하여 분석하였다. 분석 결과 표준용액의 검량선은 OTA가 결정계수 0.9999, ZEA가 0.9995로 높은 직선성을 나타냈으며, 검출 및 정량한계는 OTA가 각각 $0.1{\mu}g/kg$, $0.3{\mu}g/kg$, ZEA는 각각 $5{\mu}g/kg$, $16.7{\mu}g/kg$을 나타냈다. 유효성 및 정밀성 검정결과 OTA는 볏짚이 회수율 84.23~95.33%, 상대표준편차 (RSD) 2.59~4.77%, 이탈리안 라이그라스는 회수율 79.02~95%, RSD 0.86~5.83%, 청보리는 회수율 74.93~97%, RSD 0.85~9.19%, 호밀은 회수율 77.99~96.67%, RSD 0.33~6.26%를 나타냈으며, ZEA는 볏짚이 회수율 109.6~114.22%, RSD 0.67~7.15%, 이탈리안 라이그라스는 회수율 98.01~109.44%, RSD 1.65~4.81%, 청보리는 회수율 98~113.53%, RSD 0.25~5.85%, 호밀은 회수율 90.44~108.56%, RSD 2.5~4.66%를 나타내어 유럽연합에서 제시한 기준을 만족하였다. 따라서 본 연구에서 제시한 분석법은 국내의 볏짚 등 조사료 4종에서 OTA와 ZEA의 분석에 활용할 수 있음을 시사하였다. The objective of this study was to optimize analytical methods for ochratoxin A (OTA) and zearalenone (ZEA) in rice straw silage and winter forage crops using ultra-high performance liquid chromatography (UHPLC). Samples free of mycotoxins were spiked with $50{\mu}g/kg$, $250{\mu}g/kg$, or $500{\mu}g/kg$ of OTA and $300{\mu}g/kg$, $1500{\mu}g/kg$, or $3000{\mu}g/kg$ of ZEA. OTA and ZEA were extracted by acetonitrile and cleaned-up using an immunoaffinity column. They were then subjected to analysis with UHPLC equipped with a fluorescence detector. The correlation coefficients of calibration curves showed high linearity ($R^2{\geq_-}0.9999$ for OTA and $R^2{\geq_-}0.9995$ for ZEA). The limit of detection and quantification were $0.1{\mu}g/kg$ and $0.3{\mu}g/kg$, respectively, for OTA and $5{\mu}g/kg$ and $16.7{\mu}g/kg$, respectively, for ZEA. The recovery and relative standard deviation (RSD) of OTA were as follows: rice straw = 84.23~95.33%, 2.59~4.77%; Italian ryegrass = 79.02~95%, 0.86~5.83%; barley = 74.93~97%, 0.85~9.19%; rye = 77.99~96.67%, 0.33~6.26%. The recovery and RSD of ZEA were: rice straw = 109.6~114.22%, 0.67~7.15%; Italian ryegrass = 98.01~109.44%, 1.65~4.81%; barley = 98~113.53%, 0.25~5.85%; rye = 90.44~108.56%, 2.5~4.66%. They both satisfied the standards of European Commission criteria (EC 401-2006) for quantitative analysis. These results showed that the optimized methods could be used for mycotoxin analysis of forages.
후자린산(Fusaric acid) 생성 Fusarium 종의 신속 검출 PCR
이데레사,김소수,함현희,이수형,홍성기,류재기,Lee, Theresa,Kim, Sosoo,Busman, Mark,Proctor, Robert H.,Ham, Hyeonhui,Lee, Soohyung,Hong, Sung Kee,Ryu, Jae-Gee 한국식물병리학회 2015 식물병연구 Vol.21 No.4
후자린산은 Fusarium이 생성하는 독소로서 다른 Fusarium 독소와 함께 독성을 증진시킬 수 있다. 후자린산 독소를 특이적으로 검출하기 위해 후자린산의 생합성유전자 중 전사인자인 FUB10을 증폭하는 프라이머를 제작하였다. Fub10-f와 Fub10-r 프라이머쌍으로 PCR을 수행했을 때, F. oxysporum, F. proliferatum, F. verticillioides, F. anthophilum, F. bulbicola, F. circinatum, F. fujikuroi, F. redolens, F. sacchari, F. subglutinans, F. thapsinum에서 약 550 bp의 단일밴드가 증폭되었으며 이들은 모두 후자린산을 생성하는 것으로 알려졌다. 반면 트라이코쎄신을 생성하는 종에서는 FUB10 특이 밴드가 증폭되지 않았다. 후자린산은 푸모니신을 생성하는 종에서 함께 생성될 수 있기 때문에 FUB10 프라이머와 푸모니신 생합성유전자인 FUM1 프라이머를 이용한 multiplex PCR을 수행하였다. 그 결과 푸모니신 생성종인 F. proliferatum과 F. verticillioides에서 밴드가 모두 증폭되었으며 이는 두 가지 독소를 생성할 수 있는 종에서 동시 검출이 가능함을 시사하였다. Fusaric acid is a mycotoxin produced by species of the fungus Fusarium and can act synergistically with other Fusarium toxins. In order to develop a specific detection method for fusaric acid-producing fungus, PCR primers were designed to amplify FUB10, a transcription factor gene in fusaric acid biosynthetic gene cluster. When PCR with Fub10-f and Fub10-r was performed, a single band (~550 bp) was amplified from F. oxysporum, F. proliferatum, F. verticillioides, F. anthophilum, F. bulbicola, F. circinatum, F. fujikuroi, F. redolens, F. sacchari, F. subglutinans, and F. thapsinum, all of which were known for fusaric acid production. Whereas the FUB10 specific band was not amplified from Fusarium species known to be trichothecene producer. Because production of fusaric acid can co-occur in species that also produce fumonisin mycotoxins, we developed a multiplex PCR assay using the FUB10 primers as well as primers for the fumonisin biosynthetic gene FUM1. The assay yielded amplicons from fumonisin producers such as F. proliferatum and F. verticillioides, allowing for the simultaneous detection of species with the genetic potential to produce both types of mycotoxins.
오이와 고추생산 환경에서의 GAP 모델 개발을 위한 위해요소 조사
심원보(Won-Bo Shim),이채원(Chae-Won Lee),정명진(Myeong-Jin Jeong),김정숙(Jeong-Sook Kim),류재기(Jae-Gee Ryu),정덕화(Duck-Hwa Chung) 한국식품과학회 2014 한국식품과학회지 Vol.46 No.1
오이와 고추의 안전성에 영향을 미치는 위해요소 분석의 기초자료로 활용하기 위해 오이와 고추 재배지에서 총 72점의 시료를 채취하여 미생물학적(위생지표세균, 병원성미생물, 곰팡이) 위해요소와 화학적(중금속, 잔류농약) 위해요소를 조사하였다. 미생물학적 위해요소 중 위생지표세균과 곰팡이는 오이 재배지에서 ND-7.2와 ND-4.8 log CFU/g (g, mL, hand, or 100 cm²) 범위로 검출되었고, 고추 재배지에서는 ND-6.8와 0.4-5.3 log CFU/g (g,mL, hand, or 100 cm²) 범위로 검출되었다. 특히 대장균군은 고추재배지의 토양에서 최대 5.6 log CFU/g까지 검출되었다. 병원성 미생물은 S. aureus는 오이 농장의 장갑에서만 1.4 log CFU/100 cm²으로 검출되었고, B. cereus는 대부분의 시료에서 ND-4.8 log CFU/g (g, mL, hand, or 100 cm²) 범위로 검출되었다. L. monocytogenes, E. coli O157:H7 및 Salmonella spp.는 검출되지 않았다. 화학적 위해요소에 해당하는 중금속은 Zn, Cu, Ni, Pb 및 Hg이 국내 허용기준치 이하로 검출되었고, 잔류농약의 경우 오이에서는 모든 성분이 불검출 되었으나 고추에서 hexaconazole 성분이 0.016 mg/kg (기준치: 0.3 mg/kg)으로 검출되었다. To analyze the hazards associated with cucumber and hot pepper cultivation areas, a total of 72 samples were obtained and tested to detect the presence of biological (sanitary indicative, pathogenic bacteria and fungi) and chemical hazards (heavy metals and pesticide residues). The levels of sanitary indicative bacteria (aerobic plate counts and coliforms) and fungi were ND-7.2 and ND-4.8 log CFU/(g, mL, hand, or 100 cm²) in cucumber cultivation areas, and ND-6.8 and 0.4-5.3 log CFU/(g, mL, hand, or 100 cm²) in hot pepper cultivation areas. More specifically, the soil of hot pepper cultivation areas was contaminated with coliforms at a maximum level of 5.6 log CFU/g. Staphylococcus aureus was detected only in glove samples at a level of 1.4 log CFU/100 cm² and Bacillus cereus was detected in the majority of samples at a level of ND-4.8 log CFU/(g, mL, hand, or 100 cm²). Listeria monocytogenes, Escherichia coli O157:H7, and Salmonella spp. were not detected. Heavy metal (Zn, Cu, Ni, Pb, and Hg) chemical hazards were detected at levels lower than the regulation limit. Residual insecticides were not detected in cucumbers; however, hexaconazole was detected at a level of 0.016 mg/kg (maximum residue limit: 0.3 mg/kg) in hot peppers.