• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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• pp.130-130
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• 2003

Gummy stem blight pathogen is very difficult not only to monitor the inoculum levels prior to host infection, and also it is destructive and hard to control in field condition. We have applied RAPD technique to elucidate the genetic diversity of the genomic DNA of Didymella bryoniae and also to generate specific diagnostic DNA probe useful for identification and detection. The 40 primers produced clear bands consistently from the genomic DNA of twenty isolates of Didymella bryoniae, and two hundred seventy-three amplified fragments were produced with 40 primers. The combined data from 273 bands was analyzed by a cluster analysis using UPGMA method with an arithmetic average program of NTSYS-PC (Version 1.80) to generate a dendrogram. At the distance level of 0.7, two major RAPD groups were differentiated among 20 strains. RAPD group (RG) I included 8 isolates from watermelon except one isolate from melon. RAPD group (RG) IV included 12 isolates from squash, cucumber, watermelon and melon.. In amplification experiment with SCAR specific primer RG1F-RG1R resulted in a single band of 650bp fragment only for 8 isolates out of 20 isolates that should be designated as RAPD Group 1. However, same set of experiment done with RGIIF-RGIIR did not result in any amplified product.. Our attempts to detect intraspecific diversity of ITS region of rDNA by amplifying ITS region and 17s rDNA region for 20 isolates and restriction digestion of amplified fragment with 12 enzymes did not reveal polymorphic band. In order to develop RAPD markers for RGIV specific primer, a candidate PCR fragment( ≒1.4kb) was purified and Southern hybridized to the amplified fragment RGIV isolates. This promising candidate probe recognized only RGIV isolates

• Journal of Microbiology and Biotechnology
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• 제24권2호
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• pp.168-176
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• 2014

The morphology of filamentous fungi closely correlates with the productivity in submerged culture. Using itaconic acid (IA) production by Aspergillus terreus as a research model, the quantitative relationship between the growth form of A. terreus and IA production was investigated. IA fermentation was scaled up from shake flasks to a 7 L stirred tank bioreactor based on the quantitative relationship. Our results demonstrated the following: (1) Three morphologies of A. terreus were formed by changing the inoculum level and shape of the flask. (2) Investigation of the effects of the three morphologies on broth rheology and IA production revealed the higher yield of IA on dry cell weight (DCW, IA/DCW) and yield of glucose on DCW (consumed glucose/DCW) were achieved during clump growth of A. terreus. (3) By varying the $KH_2PO_4$ concentration and culture temperature, the relationships between clump diameter and IA production were established, demonstrating that the yield of IA on DCW ($R^2$ = 0.9809) and yield of glucose on DCW ($R^2$ = 0.9421) were closely correlated with clump diameter. The optimum clump diameter range for higher IA production was 0.40-0.50 mm. (4) When the clump diameter was controlled at 0.45 mm by manipulating the mechanical stress in a 7 L fermentor, the yield of IA on DCW and yield of glucose on DCW were increased by 25.1% and 16.3%, respectively. The results presented in this study provide a potential approach for further enhancement of metabolite production by filamentous fungi.

• 대한미생물학회지
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• 제22권1호
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• pp.1-8
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• 1987

Hantaan virus(HV) 76-118 strain was inoculated into suckling ICR mice by intra-nasal route with an inoculum of $10LD_{50}$. Mortality was 65% at the 3rd week after inoculation, but declined to 35% at the 4th week. Infectivity was determined by the measuring immuno-fluorescent antibody in sera. The peak of infectivity was 80% at the 4'th week after inoculation. Viremia was reached peak level of $1.7{\times}10^4\;PFU/ml$ by day 10. Immunofluorescent antibody and neutralizing antibody appeared by 2 weeks and 15-17 days respectively, but achieved similar titer by 35 days. By using a monoclonal antibody to HV 76-118, viral antigens were initially detected in inguinal and axillary lymph node by 2 days. Viral antigens in bone marrow and lung were delayed much more than in those of lymph node. These were similar with those of intra-peritoneal and intra-muscular route. Immune complex against IgG, IgM and C3 appeared by 16 days, 14 days, and 18 days respectively. The pattern of immunofluorescence in the basement membrane of glomeruli was diffuse membranous. Spotted pattern was also observed in the tissue stained with anti-mouse C3 antibody. By 20 days, control tissue was also shown immune complex in the glomeruli.

• 식물병연구
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• 제20권2호
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• pp.107-111
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• 2014

나선선충 접종 밀도가 토마토의 생육에 미치는 영향을 조사하고자, 시판 토마토 6품종을 직경 10-cm 토화분에 심고, 나선선충을 0, 0.02, 0.2, 2마리/g 토양 수준으로 접종하여 60일간 온실에서 재배하였다. 나선선충은 식물체의 무게나 뿌리의 무게에는 영향을 미치지 않았으나 키에는 영향을 주었는데(P < 0.05), 토양 g 당 나선선충 2마리 접종 시 포세이돈 품종은 무처리에 비하여 키가 24% 위축되었다. 나선선충을 접종하였을 때(2마리/g 토양), 가장 나선선충의 선충의 증식이 많았던 토마토 품종은 호용으로 최종 선충 밀도는 7.0마리/g 토양 이었고, 가장 나선선충의 선충의 증식이 적었던 토마토 품종은 미니흑수품종으로 최종 선충 밀도는 2.2마리/g 토양 이었다.

• Asian-Australasian Journal of Animal Sciences
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• 제28권3호
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• pp.343-350
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• 2015

Cocoa pod is among the by-products of cocoa (Theobroma cacao) plantations. The aim of this study was to apply a number of treatments in order to improve nutritional quality of cocoa pod for feeding of ruminants. Cocoa pod was subjected to different treatments, i.e. C (cocoa pod without any treatment or control), CAm (cocoa pod+1.5% urea), CMo (cocoa pod+3% molasses), CRu (cocoa pod+3% rumen content) and CPh (cocoa pod+3% molasses+Phanerochaete chrysosporium inoculum). Analysis of proximate and Van Soest's fiber fraction were performed on the respective treatments. The pods were then subjected to an in vitro digestibility evaluation by incubation in rumen fluid-buffer medium, employing a randomized complete block design (n = 3 replicates). Further, an in vivo evaluation of the pods (35% inclusion level in total mixed ration) was conducted by feeding to young Holstein steers (average body weight of $145{\pm}3.6kg$) with a $5{\times}5$ latin square design arrangement (n = 5 replicates). Each experimental period lasted for 30 d; the first 20 d was for feed adaptation, the next 3 d was for sampling of rumen liquid, and the last 7 d was for measurements of digestibility and N balance. Results revealed that lignin content was reduced significantly when cocoa pod was treated with urea, molasses, rumen content or P. chrysosporium (p<0.01) with the following order of effectiveness: CPh>CAm>CRu>CMo. Among all treatments, CAm and CPh treatments significantly improved the in vitro dry matter and organic matter digestibility (p<0.05) of cocoa pod. Average daily gain of steers receiving CAm or CPh treatment was significantly higher than that of control (p<0.01) with an increase of 105% and 92%, respectively. Such higher daily gain was concomitant with higher N retention and proportion of N retention to N intake in CAm and CPh treatments than those of control (p<0.05). It can be concluded from this study that treatment with either urea or P. chrysosporium is effective in improving the nutritive value of cocoa pod.

• 한국축산식품학회지
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• 제37권6호
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• pp.799-803
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• 2017

In the present study, centrifugation and filtration pretreatments were evaluated to decrease sample preparation time and to improve the sensitivity and specificity of multiplex polymerase chain reaction (PCR) for the detection of low levels of pathogenic Escherichia coli in various foods. Pathogenic E. coli (E. coli NCCP11142, E. coli NCCP14037, E. coli NCCP 14038, E. coli NCCP14039, and E. coli NCCP15661) was inoculated into pork, beef, and baby leafy vegetables at 1, 2, and 3 Log CFU/g. The samples were shaken 30 times (control), then centrifuged or filtered. DNA extracts from the samples were subjected to PCR using the $Powerchek^{TM}$ Diarrheal E. coli 8-plex Detection Kit. In the pork samples, no E. coli was detected in the control samples, while E. coli were detected in 100% of 3-Log CFU/g inoculated and centrifuged samples, and in 100% of 2 and 3-Log CFU/g inoculated, and filtered samples. In the beef samples, all control samples appeared to be E. coli-negative, while E. coli was detected in 50-75% of centrifuged samples, regardless of inoculated level, and in 100% of 2 and 3-Log CFU/g inoculated, and filtered samples. In baby leafy vegetables, E. coli were not detected in 25-50% of the control samples, while E. coli were detected in 0-25% of the centrifuged samples, and 75-100% of the filtered samples, depending on the inoculum amount. In conclusion, filtration pretreatment can be used to minimize sample preparation time, and improve the sensitivity and specificity of rapid detection of pathogenic E. coli in various foods.

• 한국식품과학회지
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• 제22권6호
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• pp.668-674
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• 1990

대두, 수수, 대두-수수 혼합곡물 템페를 인도네시아의 전통적인 종균(LARU ; mixed cultures of Rhizopus oligosporus)를 사용하여 제조하였다. 발효가 끝난 후 대두 템페와 대두-수수 혼합템페는 단백질과 섬유질의 양이 발효시키지 않은 대조군 시료보다 증가하였고, 지방 함량은 수수템페와 대두-수수 혼합템페의 경우 약간 증가하였다. 발효가 진행함에 따라 3가지 형태의 템페 모두가 pH, 수용성 고형분, 수용성 질소 함량이 증가한 반면 총고형분 함량에는 큰 변화가 없었다. 트립신억제제 활성(TIA)과 피트산 함량은 발효 32시간 후 감소되었다. 이는 Rhizopus oligosporus 균이 대두를 발효시키는 과정에서 트립신억제제와 피트산을 분해하는 분해능을 가지고 있음을 제시하는 결과이다. 티아민과 나이아신 함량이 3가지 형태의 템페 모두에서 증가하였다. 32시간 발효 후 대두, 수수, 대두-수수 혼합템페 모두에서 총아미노산 함량은 감소하였고 아미노산 중 리신, 발린, 티로신, 알라닌 등이 증가하였고 세린과 글루탐산 이 감소하였다.

• Journal of Microbiology and Biotechnology
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• 제22권7호
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• pp.960-966
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• 2012

The diversity and composition of yeast populations may greatly impact wine quality. This study investigated the yeast microbiota in two different types of wine fermentations: direct inoculation of a commercial starter versus pied de cuve method at an industrial scale. The pied de cuve fermentation entailed growth of the commercial inoculum used in the direct inoculation fermentation for further inoculation of additional fermentations. Yeast isolates were collected from different stages of wine fermentation and identified to the species level using Wallersterin Laboratory nutrient (WLN) agar followed by analysis of the 26S rDNA D1/D2 domain. Genetic characteristics of the Saccharomyces cerevisiae strains were assessed by a rapid PCR-based method, relying on the amplification of interdelta sequences. A total of 412 yeast colonies were obtained from all fermentations and eight different WL morphotypes were observed. Non-Saccharomyces yeast mainly appeared in the grape must and at the early stages of wine fermentation. S. cerevisiae was the dominant yeast species using both fermentation techniques. Seven distinguishing interdelta sequence patterns were found among S. cerevisiae strains, and the inoculated commercial starter, AWRI 796, dominated all stages in both direct inoculation and pied de cuve fermentations. This study revealed that S. cerevisiae was the dominant species and an inoculated starter could dominate fermentations with the pied de cuve method under controlled conditions.

• 미생물학회지
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• 제8권1호
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• pp.21-34
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• 1970

This experiment was undertaken to examine the injurious environment conditions for occuring of the virus disease, grasserie and cytoplasmic polyhedrosis in rearing of silk worms, to observe of cytoplsamic polyhedrosis diseased silkworms with histological preparation and to define the virus origin on the gattine and the disease of shrinked form after moulting (Okichijimi). The results obtained are as follows. 1) The grasserie in spring season rearing was remarkably infected in highly percent with 20.1 % in high temperature condition during 3rd to 4th instar, the high temperature during 1st to 2nd instar and 5th instar in 16.5% and 16.3%, respectively. In the fall season rearing, the disease was infected by the feeding of soft leaves plot in 5.3% and 4.8%, respectively with significant difference in 5% level, accordingly, it was thought to the nutritional condition is a factor in occuring of the disease. 2) In spring season rearing, the number ofl infected silk worms of cytoplasmic polyhedrosis was increased in the high temperautre and high humidity conditions, and in fall season rearing, order of the low temperature and high humidity plot, first feeding plot and feeded with hard leaves plot were found insome high infected ratio of the disease than control plot. 3) The occuring of cytoplasmic polyhedrosis was observed even in control rearing plot with the examining of anatomical and histological preparation in spring and fall. 4) It was found that the high diseased ratio of the gattine and disease of shrinked form after moulting in 21.8% of control and 93.2% in feeded with inocylated plot in the biosassay of inoculum. It was defined as a virus flacherie acoording to the Danaka and Shimizu's examine method.

• Journal of Microbiology and Biotechnology
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• 제33권10호
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• pp.1337-1350
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• 2023

Caproic acid is a precursor substance for the synthesis of ethyl caproate, the main flavor substance of nongxiangxing baijiu liquor. In this study, Clostridium butyricum GD1-1, a strain with high caproic acid concentration (3.86 g/l), was isolated from the storage pit mud of nongxiangxing baijiu for sequencing and analysis. The strain's genome was 3,840,048 bp in length with 4,050 open reading frames. In addition, virulence factor annotation analysis showed C. butyricum GD1-1 to be safe at the genetic level. However, the annotation results using the Kyoto Encyclopedia of Genes and Genomes Automatic Annotation Server predicted a deficiency in the strain's synthesis of alanine, methionine, and biotin. These results were confirmed by essential nutrient factor validation experiments. Furthermore, the optimized medium conditions for caproic acid concentration by strain GD1-1 were (g/l): glucose 30, NaCl 5, yeast extract 10, peptone 10, beef paste 10, sodium acetate 11, L-cysteine 0.6, biotin 0.004, starch 2, and 2.0% ethanol. The optimized fermentation conditions for caproic acid production by C. butyricum GD1-1 on a single-factor basis were: 5% inoculum volume, 35℃, pH 7, and 90% loading volume. Under optimal conditions, the caproic acid concentration of strain GD1-1 reached 5.42 g/l, which was 1.40 times higher than the initial concentration. C. butyricum GD1-1 could be further used in caproic acid production, NXXB pit mud strengthening and maintenance, and artificial pit mud preparation.