• 한국식품위생안전성학회지
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• 제37권1호
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• pp.9-14
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• 2022

본 연구에서는 가래떡에 오염된 곰팡이를 분리 동정한 후 곰팡이독소 생산 가능성을 조사하여 식품 이물 중 곰팡이에 대한 위험성을 분석하였다. 실험에 사용된 가래떡은 전라남도 소재 떡 제조업체에 곰팡이로 추정되는 이물이 발견되어 반품된 제품을 사용하였다. 곰팡이 이물을 채취한 후 Potato dextrose agar, Malt extract agar, Czapek yeast extract agar에 접종하고 25℃에서 7일간 배양하여 순수 분리 결과를 판단하였다. 순수 분리가 확인된 집락에 대해 광학현미경으로 포자의 미세구조를 관찰하여 형태학적으로 동정하였다. 유전학적 동정은 ITS primer를 이용하여 증폭된 PCR 산물의 유전자 염기서열을 분석하였다. Colony-1과 2는 각각 배지에서 동일한 성상을 나타내어 순수분리 되었음을 확인하였다. Colony-1은 출아세포가 관찰되어 효모로 판정하였다. Colony-2는 방추형의 분생자가 관찰되어 Fusarium 속 곰팡이로 판정하였다. 유전자 염기서열 분석 결과 Fusarium 속 곰팡이가 총 76건으로 나타났으며, 그중 Fusarium solani가 53건으로 가장 높게 나타났다. 형태학적 동정 결과와 유전학적 동정 결과를 바탕으로 Colony-2는 Fusarium spp.로 동정되었고 그 중 Fusarium solani로 추정되었다. Fusarium 속 곰팡이는 Nivalenol, Zearalenone, Fumonisin 등의 곰팡이독소를 생산하여 구토, 설사, 암 등을 유발한다. 이러한 결과를 종합해 볼 때 가래떡에서 분리된 Fusarium 속 곰팡이에 의한 곰팡이독소 생산 가능성은 상존하는 것으로 판단되었다. 따라서 곰팡이 이물이 보고되면 곰팡이를 분리 동정하여 곰팡이독소를 생산할 수 있는 균종인지 판단하고, 그 결과에 따라 행정조치를 강화하여야 할 것으로 판단된다.

• The Plant Pathology Journal
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• 제15권1호
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• pp.53-56
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• 1999

Fifty-three isolates of Fusarium graminearum were obtained from corn and barley samples in several provinces of Korea. Gas chromatography-mass spectrometric analysis of trichothecenes produced by these isolates revealed that 37 and 16 isolates were nivalenol (NIV)- and deoxynivalenol (DON)-chemotypes, respectively. Two hundred and seventy-five nitrate-nonutilizing (nit) mutants were obtained from the isolates. Of these mutants, 187 were identified as nit1, nit3, or NitM, but 88 could not be identified as one of these classes. The highest frequency of nit mutant was nit1 (65%), followed by nit3 (20%) and NitM (15%). Higher frequency of NitM was observed in DON-chemotypes than in NIV-chemotypes. The mutants were used for vegetative compatibility group (VCG) analysis by examining heterokaryosis using complementary mutant pairs. No heterokaryon formation was observed among all 1,248 pairwise combinations, suggesting that all isolates tested belong to different VCGs. Higher frequency of self-incompatibility was observed in NIV-chemotypes than in DON-chemotypes. These results suggest that the like-lihood of asexual genetic recombination may be very low I F. graminearum under the field condition.

• 한국미생물·생명공학회지
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• 제25권4호
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• pp.414-419
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• 1997

In order to develop an enzyme-linked immunosorbent assay (ELISA) for deoxynivalenol (DON) in com, we produced a specific monocl- onal antibody and established ELISA conditions. After the spleen cells from mice immunized with DON-bovine serum albumin conjugate were fused with S$_{p}$2/0 myeloma cells, a hybridoma cell 3G7 producing anti-DON antibody was screened by ELISA. From the standard curve of competitive direct ELISA (cdELISA) using 3G7 monoclonal antibody and DON-HRP conjugate, the detection range of DON showed 3-3,000 ng/ml (ppb). The monoclonal antibody showed some cross-reactivities against DON analogues such as 15 acetyl-DON (110%), nivalenol (5.0%), 3 acetyl-DON (1.7%), fusarenon-x (0.72%), and T-2 (0.59%). When the cdELISA was applied to the spiked coms after extracting with 60% methanol and diluting 5- fold with washing buffer, the assay recoveries of DON were 313, 163, 106, and 88.9% (av., 168%) in the levels of 200, 600, 2,000, and 6,000ng/g, respectively. For the quantitation of DON in coms, 30 samples kept under two different storage conditions of cold and room temperature were assayed by cdELISA. The mean detection concentrations were 595 (detection range, 0-2,750) and 2,448 (detection range, 0-4,500) ppb, respectively.

• The Plant Pathology Journal
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• 제26권4호
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• pp.321-327
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• 2010

We performed diagnostic PCR assays and a phylogenetic analysis using partial sequences of TEF1 (translation elongation factor-1) to determine the trichothecene chemotypes and genetic diversity of F. graminearum isolates from maize and rice samples collected in 2009 in Korea. PCR using a species-specific primer set revealed a total of 324 isolates belonging to the putative F. graminearum species complex. PCR with trichothecene chemotypespecific primers revealed that the nivalenol (NIV) chemotype was predominant among the fungal isolates from rice (95%) in all provinces examined. In contrast, the predominant chemotype among the corn isolates varied according to region. The deoxynivalenol (DON) chemotype was found more frequently (66%) than the NIV chemotype in Gangwon Province, whereas the NIV chemotype (70%) was predominant in Chungbuk Province. Phylogenetic analysis showed that all DON isolates examined were clustered into lineage 7, while the NIV isolates resided within lineage 6 (F. asiaticum). Compared with previous studies, the lineage 6 isolates in rice have been predominantly maintained in southern provinces, while the dominance of lineage 7 in maize has been evident in Gangwon at a slightly reduced level.

• 식물병연구
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• 제18권4호
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• pp.261-267
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• 2012

국내산 변질미와 저장 중 미곡에 발생하는 곰팡이로는 Aspergillus 속이 가장 많이 관찰되었으며, 분리한 곰팡이들의 일부는 아플라톡신 생성능을 보유하고 있었다. 대부분의 미곡시료에서는 Fusarium, Aspergillus, Penicillium, Alternaria 속 등의 다양한 곰팡이가 발생했으나 시료 내 아플라톡신 발생이나 오염수준은 허용기준치 이하로 미미한 반면 Fusarium 독소인 데옥시니발레놀, 니발레놀, 제랄레논, 푸모니신 등의 발생은 빈번하였다. 미곡종합처리장의 벼와 가공부산물의 경우, 곰팡이 및 곰팡이독소의 발생수준이 가공단계를 거칠수록 감소하여 백미에서 가장 낮았다. 현재, 벼의 가공부산물을 포함한 국내산 미곡의 경우, 아플라톡신 보다 Fusarium 곰팡이독소에 대한 안전관리가 더욱 필요한 것으로 판단된다.

• 식물병연구
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• 제19권4호
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• pp.259-264
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• 2013

미곡종합처리장에서 수집한 2011년산 쌀 시료 중 Fusarium 곰팡이독소의 자연발생을 조사하기 위해서, 벼, 왕겨, 현미, 청치미, 싸라기, 미강, 색채미, 백미 등의 시료를 2012년에 전국적으로 수집하였다. 371점의 시료에 대해 DON, NIV과 ZEA의 자연발생을 LC/MS로 분석하였다. 이중 색채미가 오염율과 오염도가 가장 높았고 다음이 싸라기였다. 백미는 3점이 DON, NIV에 안전한 수준으로 오염되었었지만 나머지는 곰팡이독소가 검출되지 않았다. 부산물은 백미보다 오염도나 오염율이 높았다.

• 한국균학회소식:학술대회논문집
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• 한국균학회 2016년도 춘계학술대회 및 임시총회
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• pp.33-33
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• 2016

Mycotoxins are toxic secondary metabolites produced by fungi. They can be present in where agricultural-based commodities are contaminated with toxigenic fungi. These mycotoxins cause various toxicoses in human and livestock when consumed. Small grains including corn, barley, rice or wheat are frequently contaminated with mycotoxins due to infection mainly by toxigenic Fusarium species and/or under environment favorable to fungal growth. One of the most well-known Fusarium toxin groups in cereals is trichothecenes consisting of many toxic compounds. Deoxynivalenol (DON), nivalenol (NIV), T-2 toxin, and various derivatives belong to this group. Zearalenone and fumonisin (FB) are also frequently produced by many species of the same genus. In order to monitor Korean cereals for contamination with Fusarium and other mycotoxigenic fungal species as well, barley, corn, maize, rice grains, and soybean were collected from fields at harvest or during storage for several years. The fungal colonies outgrown from the grain samples were identified based on morphological and molecular characteristics. Trichothecene chemotypes of Fusarium species or presence of FB biosynthetic gene were determined using respective diagnostic PCR to predict possible toxin production. Heavy grain contamination with fungi was detected in barley, rice and wheat. Predominant fungal genus of barley and wheat was Alternaria (up to 90%) while that of rice was Fusarium (~40%). Epicoccum also appeared frequently in barley, rice and wheat. While frequency of Fusarium species in barley and wheat was less than 20%, the genus mainly consisted of Fusarium graminearum species complex (FGSC) which known to be head blight pathogen and mycotoxin producer. Fusarium composition of rice was more diverse as FGSC, Fusarium incarnatum-equiseti species complex (FIESC), and Fusarium fujikuroi species complex (FFSC) appeared all at considerable frequencies. Prevalent fungal species of corn was FFSC (~50%), followed by FGSC (<30%). Most of FFSC isolates of corn tested appeared to be FB producer. In corn, Fusarium graminearum and DON chemotype dominate within FGSC, which was different from other cereals. Soybeans were contaminated with fungi less than other crops and Cercospora, Cladosporium, Alternaria, Fusarium etc. were detected at low frequencies (up to 14%). Other toxigenic species such as Aspergillus and Penicillium were irregularly detected at very low frequencies. Multi-year survey of small grains revealed dominant fungal species of Korea (barley, rice and wheat) is Fusarium asiaticum having NIV chemotype.

• The Plant Pathology Journal
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• 제32권5호
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• pp.407-413
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• 2016

Fusarium graminearum species complex (FGSC) causes Fusarium head blight in small grain cereals. To date, four species (F. graminearum, F. asiaticum, F. boothii, and F. meridionale ) belonging to FGSC frequently occur in Korean cereals. In addition, we first reported the occurrence of additional species (F. vorosii ) within FGSC, which was isolated from barley, corn, and rice in Korea. Phylogenetic analysis of the Fusarium isolates of this group using combined multigene sequences confirmed species identification. Moreover, the macroconidia produced by these isolates were morphologically similar to those of the F. vorosii holotype. Chemical analysis indicated that the F. vorosii isolates produced various trichothecenes such as nivalenol and deoxynivalenol with their acetyl derivatives along with zearalenone. Pathogenicity tests demonstrated that all of the F. vorosii isolates examined were pathogenic on barley, corn, and rice with variation in aggressiveness. This study is the first report of F. vorosii in Korean cereals, their pathogenicity towards barley and corn, and their ability to produce trichothecenes and zearalenone.

• The Plant Pathology Journal
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• 제38권6호
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• pp.637-645
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• 2022

Fusarium head blight (FHB) is one of the most serious diseases in barley and wheat, as it is usually accompanied by the production of harmful mycotoxins in the grains. To identify FHB-resistant breeding resources, we evaluated 60 elite germplasm accessions of barley (24) and wheat (36) for FHB and mycotoxin accumulation. Assessments were performed in a greenhouse and five heads per accession were inoculated with both Fusarium asiaticum (Fa73, nivalenol producer) and F. graminearum (Fg39, deoxynivalenol producer) strains. While the accessions varied in disease severity and mycotoxin production, four wheat and one barley showed <20% FHB severity repeatedly by both strains. Mycotoxin levels in these accessions ranged up to 3.9 mg/kg. FHB severity was generally higher in barley than in wheat, and Fa73 was more aggressive in both crops than Fg39. Fg39 itself, however, was more aggressive toward wheat and produced more mycotoxin in wheat than in barley. FHB severity by Fa73 and Fg39 were moderately correlated in both crops (r = 0.57/0.60 in barley and 0.42/0.58 in wheat). FHB severity and toxin production were also correlated in both crops, with a stronger correlation for Fa73 (r = 0.42/0.82 in barley, 0.70 in wheat) than for Fg39.

• 식물병연구
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• 제17권3호
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• pp.272-279
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• 2011

2010년산 쌀보리 43시료, 겉보리 17시료, 맥주보리 11시료, 청보리 14시료, 밀 41시료 등 총 126개 시료를 농가로부터 채집하였다. 이들 시료를 대상으로 Fusarium균 오염정도와 이들 균이 생산하는 주요 독소인 DON, NIV, ZEA의 자연발생을 조사하였다. 2010년산 맥류의 전국 평균 오염율은 23.0%로 2009년 전국 평균 오염율(Yeh 등, 2010)에 비해 10% 이상 높았다. 지역별 오염율은 전남지방의 맥류가 35.8%로 가장 높았고 전북 20.8%, 경기 20.5% 순이었다. 각 독소별 오염빈도와 오염수준을 조사한 결과, DON은 43시료(34%), 0.89 mg/kg 이었고, NIV는 107시료 (84.9%), 1.86 mg/kg, ZEA은 13시료(10.3%), 0.06 mg/kg이 었다. NIV의 오염빈도가 DON과 ZEA에 비해 높았다. 한편 40시료(31.7%)가 DON과 NIV가 중복오염되어 있었다. 맥류 종류별 곰팡이독소의 오염수준을 보면, 밀에서는 DON의 오염수준이 NIV보다 높았으며, 보리에서는 NIV 오염수준이 DON보다 높았다. 보리 종류별 NIV의 오염수준은 청보리, 맥주보리, 쌀보리, 겉보리 순으로 높았다. 독소오염에 대해 지역별로 분석하여보면 경기, 충남북의 중부지방은 DON의 오염수준이 NIV보다 높았으며, 전남북과 경남북의 남부지방은 NIV의 오염수준이 높았다. DON과 NIV 독소의 오염빈도 또한 중부지방에 비해 남부지방이 많았다. 지역별 맥류 곰팡이독소 자연발생 양상의 차이를 처음으로 보고한다.