• 한국미생물·생명공학회지
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• 제39권4호
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• pp.357-363
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• 2011

고역가 액체종국 제조 및 배양기술을 개발하기 위해, Aspergillus 속의 곰팡이(A. oryzae, A. niger)를 이용한 밀기울 첨가율 별(0, 5, 10, 15 및 20%) 배지를 이용한 액체종국을 제조하여 이들의 품질 특성을 조사하였다. 액체종국의 일반성분 및 효소활성(glucoamylase, acidic protease)을 조사한 결과, 배지에 밀기울 첨가량이 많을수록 균사체량이 많아졌을 뿐만 아니라 산도, 아미노산도도 증가하는 것을 알 수 있었고 액체종국에 첨가하는 밀기울 농도가 달라짐에 따라 각 균주의 효소활성이 바뀌는 것을 알 수 있었다. 또한, 효소활성에 따른 최적 배양시간은 곰팡이 종류에 따라 차이가 있었고 A. oryzae와 A. niger 곰팡이의 액체종국 배양은 10~15% 밀기울을 첨가하여 48~72시간 배양하는 최적조건을 규명하였다. 기존에 사용되었던 고체종국과 비교하여 밀입국제조에 종국으로써 적합한지를 구명하고자 A. oryzae와 A. niger를 사용한 고체종국을 접종한 밀누룩을 대조구로 하여 밀기울 10% 액체배지로 만든 액체종국의 접종량 별(1, 3, 5, 10%) 밀누룩의 배양시간 별(0, 15, 20, 36, 38, 40 hrs) 및 이화학적 특성과 효소활성(${\alpha}$-amylase, glucoamylase, acidic protease)변화를 비교하였다. 결론적으로 시간과 인력등의 경제적 가치가 많이 투입되는 고체종국 보다는 누룩 제조시 액체종국을 사용하는 것이 여러 가지 측면에서도 유리 하다고 여겨진다.

• 한국약용작물학회지
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• 제9권4호
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• pp.310-317
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• 2001

1. Bulb 유래 식물체 재분화는 식물생장조절물질이 첨가되지 않은 1/4MS 액체 배지에서 잎과 뿌리분화 및 생장, bulb 분화가 양호하였다. 2. 캘러스 현탁배양의 경우 2,4-D 1mg/ l 를 처리한 액체배지는 배발생 캘러스를 증가하였다. 식물생장조절제가 처리되지 않은 액체배지는 캘러스가 활성을 잃었고, 활성 감소는 salt strength가 감소함에 따라 급격히 감소하였다. 그리고 생장조절제가 처리되지 않는 액체배지에서 소수의 캘러스로부터 잎의 분화가 관찰되었다. 3. 솔나리 잎, 뿌리, 인편 중 재분화 식물체 유도는 인편이 가장 적합하며, 인편을 치상하였을 때, 잎의 분화는 MS 기본 배지에 NAA 1mg/ l 첨가와 1.5%의 sucrose 첨가한 곳에서 가장 양호하였다. 잎의 생장은 MS 기본 배지에 NAA 1mg / l를 첨가와 3.0%의 sucrose를 처리한 곳에서 관찰되었다. 뿌리의 분화 및 생장, 인편의 분화는 MS기본 배지에 NAA 1mg / l 를 첨가와 6%의 sucrose를 첨가한 배지에서 가장 양호함을 나타내었다. 4. 고체배지에서 잎의 분화는 spermidine에 비해 spermine첨가가 효과적이었고, 잎의 신장은 spermidine처리가 보다 효과적이었다. 뿌리의 분화 및 생장은 spermine처리에 의해 양호한 결과를 나타내었고, 인편의 분화는 spermidine첨가에서만이 나타났다. 액체배지에서 spermine과 spermidine 1mg/ l 를 처리하였을 때 잎, 뿌리 모두 분화하였다. Spermine 1mg/ l 를 첨가한 한 플라스크에서는 체세포 배 발생 이 관찰되었는데, 이는 2, 4-D 1mg/ l 를 처리한 액체배지에서 나타나는 것과 유사함을 나타내었다. 솔나리 인편배양에서 polyamine의 처리는 액체배지보다 고체배지가 효과적이었다. 5. 솔나리 인편배양을 통해 분화된 식물체의 토양순화 실험에서, 재분화된 식물체의 생존률은 vermiculite + perlite (1 : 1 by volume)에서 96.3%를 나타내어 가장 양호한 결과를 얻었고, 순화된 식물체의 뿌리 생장은 peat moss에서 가장 양호하였다.

• 한국약용작물학회지
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• 제15권5호
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• pp.346-351
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• 2007

An efficient somatic embryogenesis and plant regeneration protocol was developed for Schisandra chinensis Baill, using embryogenic cell suspensions and optimized media conditions. Friable embryogenic callus was induced from cotyledonary leaf and hypocotyl explants of 7 days old seedlings on MS agar medium supplemented with 1.0 to $4.0\;mg\;l^{-1}$ of 2,4-dichlorophenoxyacetic acid (2,4-D). Fast growing and well dispersed embryogenic cell suspensions were developed within two months when embryogenic calli were transferred to MS liquid medium containing $1.0\;mg\;l^{-1}\;2,4-D$. One third strength of MS medium was the best for both overall growth and development of somatic embryos in liquid culture. Over 3400 viable somatic embryos were produced from each 150 ml flask with an initial cell density of 30 mg in 30 ml medium. Germinated somatic embryos developed in liquid medium converted into plantlets after transferred to half-strength MS semi-solid medium. Approximately 90% of the converted plantlets were successfully transplanted to soil and grew into fertile plants.

• The Plant Pathology Journal
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• 제24권3호
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• pp.309-316
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• 2008

Mycelium of Ophiostoma quercus albino strain cultured in liquid culture media was harvested, lyophilized, and stored for examining biocontrol efficacy against wood discoloration by staining fungi in the laboratory and field conditions. Dry weight of mycelium grown in brown sugar yeast extract broth(BYB) showed 3.8 times higher than that grown in potato dextrose broth(PDB). The optimum culture period in BYB was 4 weeks. In vitality test of the albino strain, the lyophilized mycelium stored in liquid nitrogen($-196^{\circ}C$) or in a refrigerator($4^{\circ}C$) kept the vitality until 13 months after storage; however, the mycelium stored at room temperature lost the vitality completely after 13 months. The mycelium stored in liquid nitrogen or in a refrigerator protected wood chips from the discoloration by pretreating mycelial suspension on pine wood chips. The mycelium stored at room temperature for 7 months also showed complete protection. These results suggest that the lyophilized mycelium have a biocontrol efficacy only if it keeps the least vitality. In the field conditions, both albino strain and $Woodguard^{(R)}$(commercial chemical protectant) showed significant differences(p=0.05) in discoloration rate as compared to the non-treated control when these were treated on the wood logs of Pinus rigida. The albino strain showed better protection than $Woodguard^{(R)}$. Isolation frequency of blue stain fungi from the chips of wood logs treated with the albino strain was 0% at three months after treatment, while that treated with $Woodguard^{(R)}$ was 76.7%. In another experiment, pre-treatment of mycelial suspension on the cut surface of wood logs also showed significant protection from wood discoloration. Spraying of both albino strain on the cut surface and insecticides on the bark also showed relatively good control effects as compared to insecticide alone on the bark or nontreated control.

• Journal of Plant Biotechnology
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• 제6권4호
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• pp.241-246
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• 2004

The bulb scales and shoot sections ($7\;\cal{mm}\;\times\;15\;\cal{mm}$) of Lilium oriental hybrid 'Casablanca' were cultured to compare bulblet growth in vitro. Shoots were induced from in vitro grown bulbscales on MS medium with $1.0\;\cal{mg/L}\;BA,\;0.5\;\cal{mg/L}$ IAA, and 30 g/L sucrose. The regenerated shoots were cut into shoot sections, and cultured on MS medium with $2.0\;\cal{mg/L}\;BA,\;0.5\;\cal{mg/L}$ IAA and 30 g/L sucrose for shoot proliferation. Culture of shoot sections stimulated bulblet growth significantly than the bulb scales on MS medium with 60 g/L sucrose. However, the bulblets from shoot sections did not reach ideal size to produce stems with several leaves. Therefore, liquid medium was added into the same vessels to stimulate bulblet growth further. After shoot sections were cultured on MS medium with 60 g/L sucrose and 2 g/L activated charcoal for two months in dark, $20\;\cal{ml}$ liquid media containing various concentrations of sucrose and MS salts were added. Two months later, the added liquid medium stimulated bulblet growth remarkably as compared to bulblets grown without added liquid medium. The added $25\;\cal{ml}$ liquid medium containing 120 g/L sucrose and double strength of MS salts were the most effective for growth of in vitro bulblets. More than $94\%$ bulblets produced by this method sprouted stems with several leaves after cold treatment at $5^{\circ}C$ for three months.

• 한국미생물학회:학술대회논문집
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• 한국미생물학회 2003년도 International Meeting of the Microbiological Society of Korea
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• pp.26-33
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• 2003

We isolated and identifed culturable Arctic bacteria that have inhabited around Korean Arctic Research Station Dasan located at Ny-Alsund, Svalbard, Norway $(79^{\circ}N,\;12^{\circ}E)$. The pure colonies were inoculated into nutrient liquid media, genomic DNA was extracted, and phylogenetic analysis was performed on the basis of 16S rDNA sequences. Out of total 227 strains, 198 strains were overlapped or unidentified, and 43 bacteria were finally identified: 31 strains belonged to Pseudomonas, 7 strains Arthrobacter, two Flavobacterium sp., an Achromobacter sp., a Pedobacter sp., and a Psychrobacter sp. For isolation of diverse bacteria, we need more effective transport method than 3M petri-films, which were used for convenience of transportation that was restricted by volume. We also need to use other culture media than nutrient media. We expect these Arctic bacteria can be used for screening to develop new antibiotics or industrial enzymes that are active at low temperature.

• Journal of Microbiology and Biotechnology
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• 제9권3호
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• pp.282-291
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• 1999

Lactic acid bacteria were isolated from Kimchi and screened for bacteriocin. A total of 99 strains showed antimicrobial activity when grown on solid media, yet only 10 showed antimicrobial activity in liquid media. Strain H-559, identified as Lactococcus lactis subsp. lactis, exhibited the strongest inhibitory activity and was active against pathogenic bacteria including Listeria monocytogenes, Staphylococcus aureus, and Bacillus cereus as well as other lactic acid bacteria. The antimicrobial substance produced by L. lactis subsp. lactis H-559 was confirmed to be a bacteriocin by the treatment of $\alpha$-chymotrypsin, and protease type Ⅸ and ⅩIV. The bacteriocin activity remained stable between pH 2.0 and pH 11.0 and during heating for 10 min at $100^{\circ}C$. The bacteriocin production started in the exponential phase and stopped in the stationary phase. L. lactis subsp. lactis H-559 showed the highest bacteriocin activity at a culture temperature of $25^{\circ}C$, and an inverse relationship between the bacteriocin productivity and mean growth rate at different culture temperatures was observed. The mean growth rate and bacteriocin productivity of L. lactis subsp. lactis H-559 increased as the initial pH of the media increased.

• 한국약용작물학회지
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• 제12권5호
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• pp.366-371
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• 2004

Aconitine alkaloids produced from cell suspension cultures of Aconitum napellus for the first time. The effects of various culture conditions on cell biomass and aconitine accumulation in cell suspension cultures were investigated. Suspension cell cultures of A. napellus were established by transferring callus tissues from leaf explants onto liquid MS medium supplemented with $1\;mg/l$ NAA and $0.1\;mg/l$ kinetin. Among the culture media tested, MS medium had a pronounced effect on cell growth and aconitine accumulation. The maximum dry cell weight was obtained at inoculum size of 3 g (FCW) per flask and in MS medium supplemented with 5% sucrose after 8 weeks. The addition of salicylic acid (SA) and yeast extract (YE) in the MS medium enhanced aconitine accumulation. Using a proper combination of culture condition and supplements, aconitine content could reach 0.043% (dry weight basis), that was $2.5{\sim}3$ fold higher that detected in control cultures.

• 한국토양비료학회지
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• 제49권5호
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• pp.461-468
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• 2016

One of the important phytohormones produced by plant growth promoting bacteria is the auxin; indole-3-acetic acid (IAA), with L-tryptophan as the precursor. In this study, we focused on the investigation of optimal conditions for the production of IAA by Bacillus megaterium BM5. We investigated culturing conditions, such as incubation temperature, pH of the culture medium and incubation period, with varying media components such as inoculation volume, tryptophan concentration and carbon and nitrogen source. Besides, optimization study intended for high IAA production was carried out with fermentation parameters such as rpm and aeration. The initial yield of $42{\mu}g\;IAA\;ml^{-1}$ after 24 hr increased to $85{\mu}g\;ml^{-1}$ when 5% (v/v) of L-tryptophan was used in the culture broth. The maximum yield of $320{\mu}g\;IAA\;ml^{-1}$ was observed in trypticase soy broth (TSB) supplemented with starch and soybean meal as C and N sources with a C/N ratio of 3:1 (v/v) at $30^{\circ}C$, pH 8.0 for 48 hrs with 1.0 vvm and 250 rpm in 5 L working volume using 10 L scale fermenter. The bacterial auxin extracted from the culture broth was confirmed by thin layer chromatography and high-performance liquid chromatography and effect on plant growth was confirmed by root elongation test.

• 식물조직배양학회지
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• 제28권4호
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• pp.215-219
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• 2001

팔레높시스 대량생산시 PLB 증식에 미치는 배지와 배양환경에 대해 조사하였다. PLB 증식량은 MS배지에서 높았지만, 증식된 PLB상태는 NDM배지가 더욱 양호하였다 천연첨가 물인 CW 10% 또는 사과와 감자의 사용은 PLB 증식효율을 향상시키는 데 필수적이었다. 배지내 지지물에 따른 PLB 증신률은 gelrite가 첨가된 고체배지가 흰색과 분홍색계통 모두 액체탈지면 배지보다 증식이 양호하였다. PLB 증식시 적정 sucrose 농도는 10g/L이며 농도가 높아질수록 PLB가 유식물체로 재분화되었고 저농도 일수록 증식과 유식물체의 재분화가 모두 억제되었다. PLB증식을 위한 적정 PPF와 온도는 각각 14.3 $\mu$mol.s$^{-1}$m$^{-2}$ , $25^{\circ}C$이었다.