• Journal of Microbiology and Biotechnology
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• 제15권2호
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• pp.368-375
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• 2005

Batch cultures were carried out to optimize the exo-polysaccharide production by liquid cultures of Pleurotus ferulae. Among the various carbon sources, when $5\%$ of glucose was used, the maximum mycelial growth and exo-polysaccharide concentration reached were 8.78 g/l and 3.59 g/l, respectively. Yeast extract and polypeptone were identified as the most suitable nitrogen sources. In particular, when a mixture of $1\%$ of polypeptone and $0.8\%$ of yeast extract was used, 9.52 g/l of mycelial growth and 4.09 g/l of exo-polysaccharide were obtained. In the case of mineral sources, K$_2$HPO$_4$ and MgSO$_4$$\codt$7H$_2$0 were found to be the best mineral sources for mycelial growth and exo-polysaccharide production. Under the optimized culture conditions, the agitation speed and aeration were investigated for mycelial growth and exo­polysaccharide production in a jar fermentor. The maximum mycelial growth and exo-polysaccharide concentration at 1.5 vvm and 200 rpm obtained were 13.2 g/l and 4.95 g/l, respectively, after 10 days of culture, which were $76\%$ and $79\%$ higher than those of the basal medium. The specific growth rate was decreased with the increase of mycelial growth. However, the specific production rate of the exo-polysaccharide was proportionally increased with the specific growth rate. The proposed model profiles showed good agreement with the experimental results for the mycelial growth and exo-polysaccharide production. The specific production rate using the optimized medium was higher than that of basal medium.

• Journal of Microbiology
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• 제33권3호
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• pp.208-214
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• 1995

The effects of various 2, 4-D-degradative plasmids on the axenic growth patterns, the degradation phenotypes, and the competitiveness of different host bacteria were evaluated in liquid cultures; the organisms and plasmids used were Alcaligenes eutrophus JMP134/pJP4, Alcaligenes paradoxus/p2811, Pseudomonas pickettii/p712, pJP4, and p712 or p 2811 exhibited very different restriction fragment profiles in restriction endonuclease digests. These plasmids were transferred to the recipients (P. cepacia and Alcaligenes JMP228) at relatively high frequencies ranging from 8.9 $\times$ 10$^3$ to 1.6 $\times$ 10$^5$ per donar cell. In the axenic liquid cultures the fast-growing strains, such as P. pseudomallei/p745 and P. cepacia/pJP4, exhibited short lag periods, high specific growth rates, and high relative fitness coefficients, while the slow-growing strains, such as P. pickettii/p712 and A. paradoxus/p2811, had long lag periods, low specific growth rates, and low relative fitness coefficients. Depending on the type of plasmid containing the genes for the 2, 4-D pathway, some transconjugants exhibited intermediate grwoth patterns between the fast-growing strains and the slow-growing strains. The plasmid and plasmid-host interactions determined specific growth rate and lag time, respectively, which were shown to be principal determinants of competitiveness among the strains, but relative fitness coefficient derived from the axenic culture was not always predictive for the mixed culture condition.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2003년도 춘계 학술발표대회
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• pp.61-62
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• 2003

Clonal propagation of high-value forest trees through somatic embryogenesis (SE) has the potential to rapidly capture the benefits of breeding or genetic engineering programs and to improve raw material uniformity and quality. A major barrier to the commercialization of this technology is the low quality of the resulting embryos. Several factors limit commercialization of SE for Corsican pine, including low initiation rates, low culture survival, culture decline causing low or no embryo production, and inability of somatic embryos to fully mature, resulting in low germination and reduced vigour of somatic seedlings. The objective was to develop a Corsican pine maturation medium that would produce cotyledonary embryos capable of germination. Treatments were arranged in a completely randomized design. Data were analyzed by analysis of variance, and significant differences between treatments determined by multiple range test at P=0.05. Corsican pine (Pinus nigra var. maritima) cultures were initiated on modified !P6 medium. Modifications of the same media were used for culture multiplication and maintenance. Embryogenic cultures were maintained on the same medium semi solidified with 2.5 g/l Gelrite. A maturation medium, capable of promoting the development of Corsican pine somatic embryos that can germinate, is a combination of iP6 modified salts, 2% maltose, 13% polyethylene glycol (PEG), 5 mg!l abscisic acid (ABA), and 2.5 g/l Gelrite. After initiation and once enough tissue developed they were grown in liquid medium. Embryogenic cell suspensions were established by adding 0.951.05 g of 10- to 14-day-old semisolid-grown embryogenic tissue to 9 ml of liquid maintenance media in a 250ml Erlenmeyer flask. Cultures were then incubated in the dark at 2022$^{\circ}$C and rotated at 120 rpm. After 2.53 months on maturation medium, somatic embryos were selected that exhibited normal embryo shape. Ten embryos were placed horizontally on 20 ml of either germination medium ($\frac{2}{1}$strength Murashige and Skoog (1962) salts with 2.5 g/l activated charcoal) or same medium with copper sulphate adjusted to 0.25 mg/1 to compensate for copper adsorption by activated carbon. 2% and 4% maltose was substituted by 7.5% and 13% PEG respectively to improve the yield of the embryos. Substitution of' maltose with PEG was clearly beneficial to embryo development. When 2% of the maltose was replaced with 7.5% PEG, many embryos developed to large bullet-shaped embryos. At latter stages of development most embryos callused and stopped development. A few short, barrel-shaped cotyledonary embryos formed that were covered by callus on the sides and base. When 4% of the maltose was removed and substituted with 13% PEG, the embryos developed further, emerging from the callus and increasing yield slightly. Microscopic examination of the cultures showed differing morphologies, varying from mostly single cells or clumps to well-formed somatic embryos that resembled early zygotic embryos only liquid cultures with organized early-stag. A procedure for converting and acclimating germinants to growth in soil and greenhouse conditions is also tested. Seedling conversion and growth were highly related to the quality of the germinant at the time of planting. Germinants with larger shoots, longer, straighter hypocotyls and longer roots performed best. When mature zygotic embryos germinate the root emerges, before or coincident with the shoot. In contrast, somatic embryos germinate in reverse sequence, with the cotyledons greening first, then shoot emergence and then, much later, if at all, the appearance of the root. Somatic seedlings, produced from the maturation medium, showed 100% survival when planted in a field setting. Somatic seedlings showed normal yearly growth relative to standard seedlings from natural seed.

• 미생물학회지
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• 제45권3호
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• pp.281-285
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• 2009

본 연구에서는 객담 내 결핵균의 배양시험과 약제 감수성 시험 방법으로서 BacT/Alert 액체 배지, Ogawa 배지 및 $L\ddot{o}wenstein$-Jensen 배지 배양법을 비교하였다. 신속 액체배양시스템은 기존의 Egg-based media (Ogawa, $L\ddot{o}wenstein$-Jensen 배지)에 비해 높은 민감도와 빠른 배양으로 결핵환자의 초기 객담 검사와 약제 감수성 시험에 널리 사용되어 왔다. 객담은 N-Acetyl-L-cystine과 4% NaOH로 처리한 후 BacT/Alert 액체 배지 시스템과 Ogawa 배지에 접종하고 배양하여 비교하였다. 결핵환자 객담 검체 135개 중 도말검사결과 양성은 95개였고, Ogawa 배지 배양결과 양성으로 확인된 수는 89개(65.9%)였으며, BacT/Alert 액체 배지에서는 97개(71.9%)가 양성으로 확인되었다. 결핵균 배양 양성 검체의 평균배양일은 Ogawa 배지에서 22.4일, BacT/Alert 액체 배지는 11.3일로 단축되는 것을 확인하였다. 약제 감수성 시험에는 32개 결핵균주를 이용하였으며, isoniazid를 첨가한 BacT/Alert 액체 배지와 $L\ddot{o}wenstein$-Jensen 배지에서의 약제 감수성 시험 검사결과 일치률은 87.5%를 보였고, rifampicin에서의 일치률은 90.6%로 나타났다.

• 한국작물학회지
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• 제51권spc1호
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• pp.247-250
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• 2006

본 연구는 polyamine이 쪽 모상근 배양에서 뿌리의 생장과 indigo 생합성에 미치는 영향을 조사하였다. 1. 토양미생물인 Agrobacterium rhizogenes R1000을 이용하여 쪽 자엽으로부터 모상근을 효과적으로 유도하여 배양하였다. 2. 모상근 생장과 인디고 생합성이 polyamines(putrescine, spaermidine, spermine) 처리에 의하여 향상되었으며, 그 중 putrescine이 다른 polyamines에 비하여 효과적이었다. 3. Putrescine 70 mg/l 처리가 모상근의 생육(4.4 g/flask)과 인디고 생합성(216.3 ug/g)의 향상에 최적조건으로 조사되었다.

• 미생물학회지
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• 제30권2호
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• pp.96-100
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• 1992

토양세균의 혼합배양에 의한 MCPP [2-(2-Methyl-4-Chlorophenoxy) Propionic Acid]의 미생물학적 분해를 조사하였다. 혼합배양은 Pseudomonas 종, Flavobacterium 종, 그리고 Achromobacter종을 포함하였다. 세균들은 유일한 탄소 및 에너지원으로서 MCPP를 이용하였으나, 부분적인 분해만을 부여주었다. MCPP의 분해는 2-methyl-4-chlorophenol의 형성에 의하여 진행되었으며, 이는 HPLC 에 의하여 발견되었고, GC-MS에 의하여 확인되었다. 이 중간대사물질은 일시적으로 생성되었으며, reverse phase HPLC와 Uv absorbance에 의하여 다른 대사산물의 존재는 관찰되지 않았다.

• 생약학회지
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• 제26권4호
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• pp.419-425
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• 1995

Corydalis remota Fish. ex Max. (Papaveraceae) is a well known medicinal plant being used as analgesics or anticonvulsive in oriental medicine. As the alkaloid content is known to vary depending on the environmental factors, the technology of plant tissue culture can be adopted as source of Corydalis-alkaloids. The present study describes an establishment of tissue cultures of Corydalis which produce alkaloids consistently. Callus were induced from immature seeds of Corydalis remota by placing the seeds on MS static media containing NAA(0.25, 1.0 and 4.0 mg/l, respectively). The combined treatment of NAA(1.0 mg/l) with cytokinin(BAP 0.5 mg/l) improved the induction of callus. TLC scanning data followed by sequential extraction and purification revealed that the induced callus contains a significant amount of alkaloids. Cell suspension cultures were established by transferring the induced callus into the liquid media with the same condition of plant growth regulators as the callus culture.

• KSBB Journal
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• 제6권1호
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• pp.1-7
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• 1991

To produce economically important indole alkaloids by cell cultures, we selected protoplastsderived clones (protoclones) of vinca (Catharanthus roseus) for high yields of catharanthine and ajmalicine. Protoplasts were enzymatically isolated from suspension-cultured cells. The highest plating efficiency (1%) was obtained when protoplasts were plated at a density of 1$\times$105 protoplasts/ml in a culture medium solidified with 0.4% Seaplaque agarose. The growth rates of 40 protoclones subcultured on a solid medium varied over a wide range. Protoclone VPC-6, which had the highest growth rate, was observed to produce relatively high yields of catharanthine and ajmalicine when cultured in a liquid medium. Although the original cell line did not produce catharanthine at a detectable level by HPLC, protoclone VPC-10 produced it at a level of 5.9$\mu\textrm{g}$/g fresh weight of cells for 10 days of culture. Under the same conditions, protoclone VPC-15 produced ajmalicine at a level of 133.6$\mu\textrm{g}$/g, of which productivity was improved about ,3 times than that of the original cell line. The results indicate that differences in the growth rate and indole alkaloid yield among the protoclones reflect the somaclonal variation in suspnsion-cultured cells.

• 한국식물병리학회지
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• 제14권6호
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• pp.700-704
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• 1998

A total of 26 and 55 isolates of fungi were isolated from corn and wheat samples collected from different markets in Korea, respectively. The number of Penicillium isolates from corn and wheat was 9 and 33, respectively. The Penicillium species isolated from corn were P. chrysogenum (3 isolates) and P. oxalicum (6 isolates), and from wheat were P. aurantiogriseum (16 isolates), P. citrinum (1 isolate), P. commun (4 isolates), P. griseofulvum (1 isolate), P. verrucosum (7 isolates), and P. viridicatum (4 isolates). Production of major mycotoxins in the yeast extract sucrose medium cultures of Penicillium isolates was analysed. Penicillium cultures were extracted with chloroform and purified by thin-layer chromatograhy (TLC), and high performance liquid chromatography (HPLC). Among 9 isolates of Penicillium from corn, 2 isolates of P. chrysogenum produced patulin, 1 isolate of the fungus produced patulin and citrinin, 2 isolates of P. oxalicum produced penicillic acid, 4 isolates produced pencillic acid and griseofulvin. Of the 33 isolates of Penicillium from wheat, 6 isolates of P. aurantiogriseum produced patulin, 8 isolates produced penicillic acid, 1 isolate produced patulin and penicillic acid, 1 isolate of P. citrinum produced citrinin and patulin, 2 isolates of P. commun produced brefeldin A and patulin, 1 isolate of P. griseofulvum produced brefeldin A, griseofulvin and patulin. Five isolates of P. verrucosum produced patulin, 1 isolate of the fungus produced penicillic acid, and 3 isolates of P. viridicatium produced penicillic acid.

• Journal of Plant Biotechnology
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• 제5권2호
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• pp.131-135
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• 2003

Callus and cell suspension cultures of Eurycoma longifolia Jack were initiated from leaves of different trees. The leaf explant of tree Eu9 produced the most calli and also induced high cell biomass in the cell suspension culture. Optimum production of cell biomass could be initiated in proliferating culture medium with a pH of 5.75 prior to autoclaving. The effects of macronutrient inorganic salts of Murashige and Skoog (MS) liquid medium supplemented with X on production of cell biomass of Eurycoma longifolia were also investigated. The highest cell biomass was produced in MS medium containing macronutrients of $21\;mM\;NH_4NO_3,\;12.25\;mM\;KNO_3,\;3.00\;mM\;CaCl_2.2H_2O,\;0.575\;mM\;MgSO_4.7H_2O$, and $1.83\;mM\;KH_2PO_4$. A new medium labeled as TAM was formulated for the production of Eurycoma longifolia cell biomass in the cell suspension culture.