• Title/Summary/Keyword: Liquid cultures

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Establishment and Characterization of Clonal Cell Lines from Zebrafish, Danio rerio (제브라피쉬(Danio rerio) 배아로부터 동형세포주 확립)

  • Lee, Ki-Young
    • Korean Journal of Ichthyology
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    • v.20 no.1
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    • pp.1-6
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    • 2008
  • Three types of clonal cell lines were isolated according to their size and phenotype from the adherent cell populations in long-term liquid cultures from the embryonic fibroblast cells of Zebrafish, Danio rerio. All kind of cell lines were well proliferated. The size and number of clonal cell lines derived colonies from stable embryonic cells were significantly increased in the presence of NAC and A2P conditioned medium from the cell lines. The stable cell lines and clonal cell lines were cap-able of well proliferation in vitro. These cell lines have been maintained in continuous culture without change in characteristics. A majority of the clonal cells (80%) was shown a normal chromosomal complement (50 chromosomes, 2N) in according with FACs analysis. Majority of cells were positive to vimentin staining and none of them were positive for nestin and Oct -4 by immunocytochemistry. These results indicate that the clonal cell lines obtained from cultured cells are fibroblasts and may be extremely useful in genetic manipulation for further nuclear transfer and fish cloning.

Antifibrotic Effect of Extracellular Biopolymer from Submerged Mycelial Cultures of Cordyceps militaris on Liver Fibrosis Induced by Bile Duct Ligation and Scission in Rats

  • Nan, Ji-Xing;Park, Eun-Jeon;Yang, Byung-Keun;Song, Chi-Hyun;Ko, Geonil;Sohn, Dong-Hwan
    • Archives of Pharmacal Research
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    • v.24 no.4
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    • pp.327-332
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    • 2001
  • The antifibrotic effects of hot water extract (WEC), intracellular biopolymer (IPC) and extracellular biopolymers (EPC) from mycelial liquid culture of Cordyceps militaris on liver fibrosis were studied. Liver fibrosis was induced by a bile duct ligation and scission (BDL/S) operation, duration of 4 weeks in rats. In BDL/S rats, the levels of aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (ALP), total bilirubin in serum and hydroxyproline content in liver were dramatically increased. The WEC or IPC treatment (30mg/kg/day for 4 weeks, p.o.) in BDL/S rats reduced the serum AST, ALT and ALP levels significantly (p<0.01). The EPC treatment (30 mg/kg /day for 4 weeks, p.o.) reduced the serum ALT, AST and ALP levels significantly (p<0.01). Malondialdehyde contents in liver treated with WEC, IPC or EPC were significantly reduced (p <0.05). But Liver hydroxyproline content was decreased only in EPC treated BDL/S rats to 55% that of BDL/S control rats (p < 0.01). The morphological characteristics and expression of alpha smooth muscle like actin in fibrotic liver, which appeared in BDL/S control group were improved in EPC treated fibrotic liver. These results indicate that IPC (30 mg/kg /day for 4 weeks, p.o.) has an antifibrotic effect on fibrotic rats induced by BDL/S.

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The Plant Growth-Promoting Fungus Aspergillus ustus Promotes Growth and Induces Resistance Against Different Lifestyle Pathogens in Arabidopsis thaliana

  • Salas-Marina, Miguel Angel;Silva-Flores, Miguel Angel;Cervantes-Badillo, Mayte Guadalupe;Rosales-Saavedra, Maria Teresa;Islas-Osuna, Maria Auxiliadora;Casas-Flores, Sergio
    • Journal of Microbiology and Biotechnology
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    • v.21 no.7
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    • pp.686-696
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    • 2011
  • To deal with pathogens, plants have evolved sophisticated mechanisms including constitutive and induced defense mechanisms. Phytohormones play important roles in plant growth and development, as well as in the systemic response induced by beneficial and pathogen microorganisms. In this work, we identified an Aspergillus ustus isolate that promotes growth and induces developmental changes in Solanum tuberosum and Arabidopsis thaliana. A. ustus inoculation on A. thaliana and S. tuberosum roots induced an increase in shoot and root growth, and lateral root and root hair numbers. Assays performed on Arabidopsis lines to measure reporter gene expression of auxin-induced/ repressed or cell cycle controlled genes (DR5 and CycB1, respectively) showed enhanced GUS activity, when compared with mock-inoculated seedlings. To determine the contribution of phytohormone signaling pathways in the effect elicited by A. ustus, we evaluated the response of a collection of hormone mutants of Arabidopsis defective in auxin, ethylene, cytokinin, or abscisic acid signaling to the inoculation with this fungus. All mutant lines inoculated with A. ustus showed increased biomass production, suggesting that these genes are not required to respond to this fungus. Moreover, we demonstrated that A. ustus synthesizes auxins and gibberellins in liquid cultures. In addition, A. ustus induced systemic resistance against the necrotrophic fungus Botrytis cinerea and the hemibiotrophic bacterium Pseudomonas syringae DC3000, probably through the induction of the expression of salicylic acid, jasmonic acid/ethylene, and camalexin defense-related genes in Arabidopsis.

Phytotoxicity of dehydrofusaric acid isolated from Fusarium oxysporum against several plants (Fusarium oxysporum이 생산하는 dehydrofusaric acid의 몇 가지 식물 생육에 대한 활성)

  • Hong, Kyung-Sik;Choi, Gyung-Ja;Kim, Heung-Tae;Hwang, In-Taek;Cho, Kwang-Yun;Kim, Jin-Cheol
    • The Korean Journal of Pesticide Science
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    • v.6 no.3
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    • pp.202-208
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    • 2002
  • A phytotoxin was purified by repeated chromatography from liquid cultures of Fusarium oxysporum BG isolated from barnyardgrass. Its chemical structure was determined to be dehydrofusaric acid by mass and NMR spectral analyses. The substance showed a potent phytotoxic activity against growth of duckweed with a $EC_{50}$ value of $1.5{\mu}g/ml$. It also inhibited the root growth of barnyard millet, cress, barnyard grass, and rice cultivar 'Dongjin'. However, it had no inhibitory activity against seed germination of barnyard millet and cress, and the shoot growth of the four plant species.

Growth and Production of Insecticidal Crystal Proteins of Bacillus thuringiensis as Affected by Carbon Sources (Bacillus thuringiensis 생장과 살충성 결정단백질 생성에 대한 탄소원의 영향)

  • Kim, Moo-Key;Ahn, Byung-Koo
    • Applied Biological Chemistry
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    • v.39 no.3
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    • pp.177-182
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    • 1996
  • Effects of 14 carbohydrates supplied as carbon sources on cell growth and sporulation of, and the production of insecticidal crystal proteins by Bacillus thuringiensis strains were investigated in liquid cultures. Strains grew well in media containing any one of the 14 carbohydrates supplied, reaching maximum cell densities of $10^7{\sim}10^8\;cells/ml$ in 16.7 to 22 hours after inoculation depending on the strain. Spores first appeared in 16.7 to 24.7 hours after inoculation, and 80% sporulation was reached in 28 to 51.3 hours after inoculation depending on the strain. No change in pH of media was observed after cell multiplication. The production of total protein was highest when supplied with sucrose and was lowest with starch. More insecticidal crystal proteins were produced when supplied with glucose, lactose, maltose, or sucrose. The amount of insecticidal crystal proteins produced by the strains was proportional to that of the total protein. The relative amount of individual insecticidal crystal protein species produced by B.t. kurstaki and B.t. israelensis was not influenced by the carbohydrates supplied.

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Effect of Developmental Stage, Temperature, and Medium on Embryogenesis in Anther Cultures of Chinese Cabbage (Brassica campestris ssp. pekinensis) (배추(Brassica campestris ssp. pekinensis ) 약 배양시 약의 발달 단계, 온도 처리 및 배지의 종류가 배 발생에 미치는 영향)

  • 김태일;황주광;백기엽
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.4
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    • pp.229-234
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    • 1995
  • Anthers of hybrids and inbred lines of Chinese cabbage (Brassica campestris ssp. pekinensis) were inoculated on the modified solid or liquid B$_{5}$ medium after several pre-treatments. Anthers were then maintained at 25 $^{\circ}C$ after being subjected to various post-treatment. Somatic embryos began to appear 9 days after inoculation and ended at 13th days. Low temperature pre-treatment did not increase embryoid production whereas high post-treatment temperature of 4$0^{\circ}C$ or 45$^{\circ}C$ enhance the production. There were considerable levels differences in embryogenesis between lines used, but not between the culture methods. Somatic embryo yield was also increased by subjecting anthers to one day at 35$^{\circ}C$ and then another one day at 3$0^{\circ}C$ after plating. Histological observations showed several stages in haploid development ranging from a few celled to large multiple-celled embryoids.s.

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Isolation and Characterization of Burkholderia cepacia EB215, an Endophytic Bacterium Showing a Potent Antifungal Activity Against Colletotrichum Species (탄저병균에 길항력이 우수한 식물내생세균 Burkholderia cepacia EB215의 분리 및 특성 규명)

  • Park Ji Hyun;Choi Gyung Ja;Lee Seon-Woo;Jang Kyoung Soo;Lim He Kyoung;Chung Young Ryun;Cho Kwang Yun;Kim Jin-Cheol
    • Microbiology and Biotechnology Letters
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    • v.33 no.1
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    • pp.16-23
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    • 2005
  • In order to develop a new microbial fungicide using endophytic bacteria for the control of anthracnoses occurring on various crops, a total of 260 bacterial strains were isolated from fresh tissues of 5 plant species. After they were cultured in broth medium, their antifungal activities were tested for in vivo antifungal activity against cucumber anthracnose caused by Colletotrichum orbiculare. As the results, liquid cultures of 28 strains showed potent antifungal activities more than $90\%$ against cucumber anthracnose. At 3-fold dilutions of liquid cultures, 18 strains inhibited the development of cucumber anthracnose of more than $70\%$. They were further tested for in vivo antifungal activity against red pepper anthracnose caused by C. coccodes and in vitro antifungal activity against C. acutatum, a fungal agent causing red pepper anthracnose. Among 18 strains, a bacterial strain EB215 isolated from cucumber roots displayed the most potent antifungal activity against Colletotrichum species. It was identified as Burkholderia cepacia based on its physiological and biochemical characteristics, Biolog test and 16S rDNA gene sequence. It also controlled effectively the development of rice blast (Magnaporthe grisea), rice sheath blight (Corticium sasaki), tomato gray mold (Botrytis cinerea), and tomato late blight (Phytophthora infestans). Studies on the characterization of antifungal substances produced by B. cepacia EB215 are in progress.

Effects of Liquid Culture of Agaricus blazei Murill on Lipid Metabolism and Enzyme Activities in Rats Fed High Fat Diet (신령버섯 균사체 액체배양액이 고지방 식이를 급여한 흰쥐의 지질대사 및 효소활성에 미치는 영향)

  • 이현정;고진복
    • Journal of Nutrition and Health
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    • v.36 no.4
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    • pp.352-358
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    • 2003
  • This study was performed to investigate the effects of liquid culture of Agaricus blazei on the lipid metabolism and enzyme activities in growing male rats. Sprague-Dawley rats were given four different types of diets for a succeeding period of 5 weeks, respectively: a normal diet group (7% corn oil), a high fat diet group (7% com oil + 15% lard), a 20 or 30% Agaricus diet groups (high fat diet + 20 or 30% Agaricus in water) according to the levels of Agaricus supplementation. The body weight gains, food intake, food efficiency ratios, and hepatic, kidney, spleen and pancreas weights of the rats fed 20 or 30% Agaricus diets were similar to those of the rats fed high fat diet. The epididymal fat pad weight of the rats fed high fat diet and 20 or 30% Agaricus diets were significantly higher than that of the rats fed normal diet. The concentrations of serum triglyceride, total cholesterol, LDL-cholesterol and HDL-cholesterol, and the activity of glutamic pyruvic transaminase in the rats fed 30% Agaricus diet were significantly lower than those in the rats 114 high fat diet. But the concentrations of hepatic total cholesterol and triglyceride of rats fed the 20 or 30% Agaricus diets were similar to those of rats fed the high fat diet. The HDL-oholesterol/total-cholesterol ratio of the rat fed 30% Agaricus diet was significantly higher than that of the rats fed high fat diet, The activity of glutamic oxaloacetic transaminase in the rats fed 20 or 30% Agaricus diets were similar to those in the rats fed high fat diet. There were no differences in the concentrations of total protein, hemoglobin and glucose, and the activities of alkaline phosphatase, and the atherogenic index in the serum among the experimental groups. These results showed that the 30% Agaricus diet feeding decreased the total cholesterol, the triglyceride and the LDL-cholesterol, and increased the HDL-oholesterol/total cholesterol ratio in serum of rats.

Quality characteristics of commercial liquid type yogurt in Korea (국내 시판 액상요구르트의 품질특성)

  • Lee, Ji Hyun;Park, Hye Young;Won, Ju In;Park, Hye In;Choi, In Duck;Lee, Seuk Ki;Park, Ji Young;Joe, Dong Hwa;Jeon, Yong Hee;Oh, Sea Kwan;Han, Sang Ik;Choi, Hye Sun
    • Food Science and Preservation
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    • v.24 no.6
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    • pp.865-870
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    • 2017
  • Yogurt is produced by fermentation of milk using bacteria known as "yogurt cultures". Most of these bacteria are probiotics such as Lactobacillus delbrueckii subsp. bulgaricus, Lb. rhamnosus, Streptococcus thermophilus, and Bifidobacterium. The domestic fermented milk market is increasing, and about 30 companies are producing yogurt. The purpose of this study was to analyze the quality characteristics of domestic commercial liquid yogurt. Total 30types of commercial yogurts were sampled and their physicochemicial properties, including pH, sugar content, acidity, viscosity, and microbial characteristics of lactic acid bacteria counts, were measured. Commercial liquid type yogurt showed a pH of 4.5, sugar content of 7.4-21.2%, total acid content of 0.4-0.9%, and viscosity of 0.1-250 cP. In terms of microbial populations, lactic acid bacteria counts were 7.2-11.3 log CFU/mL and anaerobic lactic acid bacteria counts were 8.0-11.5 log CFU/mL. The quality characteristics differed depending on the constituents of the sample and the microorganisms used. These results are related to the quality characteristics of yogurts and are useful for identifying new trends in the domestic fermented milk industry.

Effect of Scrapping Aerial Mycelia and Light on the Production of Macroconidia and Chlamydospores of Cylindrocarpon destructans Causing Root Rot of Panax ginseng (기중균사 제거와 광처리가 인삼 뿌리썩음병균 Cylindrocarpon destructans의 대형분생포자 및 후막포자 생성에 미치는 영향)

  • Cho Dae-Hui;Yu Yun-Hyun;Ohh Seung-Hwan
    • Journal of Ginseng Research
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    • v.23 no.3 s.55
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    • pp.123-129
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    • 1999
  • Under the light condition of 25,000 Lux (12 hrs dark and light cycle) with scrapping treatment of aerial mycelia of Cylindrocarpon destructans on potato dextrose agar (PDA), V-8 juice agar, and ginseng extract agar, production of the macroconidia was increased to $3.7\~8.1$ fold over them produced in the dark. They were also produced $7.7\~18.0$ times more in the liquid cultures under the light condition than under the dark as well. PDA and V-8 juice agar among the tested were the best for the macroconidium production. On PDA, 1,585 $macroconidia/mm^2$ were produced under the light of 25,000 Lux with scrapping treatment of aerial mycelia of C. destructans, which is 3.2 and 1.4 times more than those produced under 3,000 and 10,000 Lux, respectively. Meanwhile, $20\~99$ macroconidia/$mm^2$ were produced by the non-scrapping under the light condition between 3,000 Lux and 25,000 Lux. The macroconidia were, however, lysed at $6\~7$ days after being incubated under the above range of the light. They were consisted of $1\~3$ cells in a macroconidium while $69.4\~100\%$ of them were the two-celled and the number did not seem to be affected by either the scrapping or the light. Production of chlamydospore converted from mycelia of C. destructans seemed to be promoted by the light and the scrapping as well. The 1,285 chlamydospres/$mm^2$ were produced with the light (25,000 Lux), which is 2.8 and 1.2 times more than those with 3,000 and 10,000 Lux, respectively. Scrapping the aerial mycelia of the cultures increased the chlamydospore formation to 1.9, 2.5 and 1.4 times more than the non-scrapping under the light intensity of 3,000 Lux, 10,000 Lux, and 25,000 Lux, respectively. On PDA, 1 to 8 chlamydospore(s) per catena were formed by all treatments tested and $34.2\~58.9\%$ of them was a single chlamydospore, However, the numbers was affected by neither the light ($3,000\~25,000$ Lux) nor the scrapping the aerial mycelia.

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