• Title/Summary/Keyword: minimal medium

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Detection of Auxotrophic Mutants form Valsa ceratosperma, the Causal Fungus of Apple Canker (사과나무 부란병균(腐爛病菌) Valsa ceratosperma에서의 Auxotrophic Mutants의 검출(檢出))

  • Hong, Yeon Gyu;Uhm, Jae Youl
    • Current Research on Agriculture and Life Sciences
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    • v.5
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    • pp.119-126
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    • 1987
  • This study was conducted to elucidate the most appropriate method to obtain auxotrophic mutants from Valsa ceratosperma, the causal fungus of apple canker, which may be used as a gene marker in detecting the transfer of the factors of avirulent strains to virulent strains. Among the 3 kinds of synthetic media tested, each have two formula for minimal and complete, the medium which has been used in study of Endothia parasitica (E. P medium) was turned out to be most appropriate for the growth of V. ceratosperma. A medium for single colony formation from pycnidiospore of this fungus was developed by adding 0.5% L - sorbose to the E. P minimal medium. The period of incubation in dark for preventing the photoreactivation after U. V irradiation was estimated as about 60hrs at which most of the spores become binucleate. Largest number of putative auxotrophs were obtained at about 50second of irradiation to the spores smeared on the medium for single colony formation, at which the survival rate of spores was 5 to 6 percent. With these method developed in this experiment, 161 isolates of putative auxotrophs were detected among which the nutrient requirement for 10 isolates were determined. Five out of 10 mutants were still virulent to apple tree and all but one could not sporulate.

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The Effect of Fibroblast Co-culture on In Vitro Maturation of Mouse Preantral Follicles

  • Kim, Chung-Hoon;Cheon, Yong-Pil;Lee, You-Jeong;Lee, Kyung-Hee;Kim, Sung-Hoon;Chae, Hee-Dong;Kang, Byung-Moon
    • Development and Reproduction
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    • v.17 no.3
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    • pp.269-274
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    • 2013
  • This study was performed to evaluate the effects of fibroblast co-culture on in vitro maturation (IVM) of prepubertal mouse preantral follicles. The intact preantral follicles were obtained from the ovaries of 12-14 day old mice and these were cultured individually in ${\alpha}$-minimal essential medium (${\alpha}$-MEM) supplemented with 5% fetal bovine serum (FBS), $100mIU/m{\ell}$ recombinant follicle stimulating hormone (rFSH), 1% insulin-transferrin-selenium, $100{\mu}g/ml$ penicillin and $50{\mu}g/m{\ell}$ streptomycin as base medium for 12 days. A total of 200 follicles were cultured in base medium co-cultured with mouse embryonic fibroblast (MEF) (MEF group) (n=100) or only base medium as control group (n=100). Survival rate of follicles on day 12 of culture were significantly higher in the MEF group of 90.0%, compared with 77.0% of the control group (p=0.021). Follicle diameters on day 6 and 8 of the culture period were significantly larger in the MEF group than those in the control group (p=0.021, p=0.007, respectively). Estradiol levels in culture media on day 4, 6, 8, 10 and 12 of the culture period were significantly higher in the MEF group (p=0.043, p=0.021, p=0.006, p<0.001 and p=0.008, retrospectively). Our data suggest that MEF cell co-culture on IVM of mouse preantral follicle increases survival rate and promotes follicular growth and steroid production.

Effect of Culture Media on Embryonic Cell Growth in Zebrafish, Danio rerio (배지에 따른 제브라피쉬(Danio rerio) 배아 유래세포의 성장 효과에 관한 연구)

  • Lee, Ki-Young;Kim, Jong-Yeon;Jo, Soo-Gun
    • Development and Reproduction
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    • v.12 no.1
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    • pp.51-56
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    • 2008
  • To optimize the cell culture conditions of zebrafish embryonic cells, we compared the efficiency of three types of medium, DMEM, K-NAC and D-NAC. In this study, we showed that the cells grown in K-NAC have better plating efficiency than DMEM, especially in the case of low cell seeding density. However, cells grew slower in K-NAC than those in DMEM in confluent cultures. The effect of 0.1% zebrafish embryo extracts was minimal. The presence of 1% trout serum in culture medium significantly increased the growth rate of cells(p<0.05). No difference was found at $2{\sim}3{\times}10^5$ cell seeding density(p<0.05). At $4{\sim}5{\times}10^5$ cell seeding density, cells grew better in DMEM than K-NAC (p<0.1). The results suggest that supplementation of NAC and A2P in Keratinocyte SFM may improves plating efficiency when cells are plated at low population. No difference was found for cell growth in either medium with 5%, 10% or 15% FBS supplemented (p<0.05). Cells culture in D-NAC grew significantly better than those in DMEM(p<0.05). Our results clearly showed that the use of NAC and A2P in the culture medium has a positive effect on cell growth regardless of the amount of FBS added.

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High Frequency Plant Regeneration from Leaf Explant Cultures of Domestic Cultivated Strawberry (Fragaria x ananassa Duch) (국내 딸기 재배품종의 잎절편 배양으로부터 고빈도 식물체 재생)

  • Cho Mi-Ae;Choi Kyu-Myeong;Ko Suck-Min;Min Sung-Ran;Chung Hwa-Ji;Liu Jang-Ryol;Choi Pil-Son
    • Journal of Plant Biotechnology
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    • v.32 no.3
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    • pp.175-179
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    • 2005
  • To develop a high efficiency plant regeneration system from in vitro cultures of strawberry, cv. Yeobong, petiole and leaf explants were cultured on MS basal medium containing a combination of 0.5 mg/L IBA and 3.2 mg/L kinetin or zeatin or benzyl amino purine (BAP) for 6 weeks, and leaf explants with dark pretreatment for a week ($T_1$), 2 weeks ($T_2$), and 4 weeks ($T_3$) were cultured on medium supplemented with 0.5 mg/L IBA and 3.2 mg/L zeatin under 16 hr photoperiod for 6 weeks. Shoot organogenesis was observed from the greenish calli containing minimal anthocyanin formed at proximal cutting edges of the leaf explant (57%) when cultured adaxial side on the medium, whereas was directly formed from a cutting edges of petiole explant (6.3%). Frequency of callus formation and shoot organogenesis at large size of leaf explant ($1.0{\sim}1.5\;cm^2$) was higher than small size ($0.5{\sim}1.0\;cm^2$), and dark pretreatment significantly improved the frequency of leaf explants that produced calli and shoots. The maximum frequency (87%) for shoot organogenesis was obtained from the leaf explants that transferred to a 16 hr photoperiod condition after the initial 4 weeks dark period. The improved frequency (87%) in comparision with control without dark pretreatment (27%). When the shoots were transferred to 1/2 MS basal medium, formed roots with 20 d of culture. The rooted plants were subsequently transferred to the pots and to the field.

Effect of oil and aqueous extract of Neem (Azadirachta indica) seeds on growth of Aspergillus species and biosynthesis of aflatoxin

  • Rashid, Faraz;Naaz, Farah;Abdin, MZ;Zafar, Shadab;Javed, Saleem
    • Advances in Traditional Medicine
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    • v.5 no.4
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    • pp.308-315
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    • 2005
  • Aflatoxin contamination is a major problem in several food crops. Aflatoxin, a mycotoxin, produced by Aspergillus flavus has gained immense concern in the scientific world because of its tremendous harmful effects. The study was focused to see the effect of oil and aqueous extract of neem (Azadirachta indica) seeds on the growth of Aspergillus and production of aflatoxin by the mold. Various amounts of neem oil $(5\;-\;50\;{\mu}l/ml)$ and aqueous extract of neem (5 - 50 mg/ml) were used both in the broth as well as the solid medium. Fungistatic (MIC) and minimal fungicidal concentrations (MFC) were found to be $10\;{\mu}l/ml$ and $50\;{\mu}l/ml$ respectively for neem seed oil. At the concentration of $5\;{\mu}l/ml$ neem oil and 5 mg/ml of aqueous extract, a significant decrease in the aflatoxin content was found in broth medium. Aflatoxin production was totally inhibited at $50\;{\mu}l/ml$ and 50 mg/ml for neem oil and aqueous extract of neem respectively, in both treatments. There was significant inhibition of mycelium dry weight by the neem seed oil. Mycelial growth was totally inhibited at $20\;{\mu}l/ml$ of neem seed oil concentration in broth, whereas it was not affected at all by aqueous extract. It can therefore be inferred that the oil and extract from the neem seed leads to inhibition of aflatoxin production while neem seed oil also significantly inhibits the mycelial growth. Neem seed oil thus can be used as potent, natural and easily available anti-aflatoxigenic agent.

The Cultural Characteristics and Antibacterial Activities of Cordyceps militaris and Paecilomyces tenuipes (번데기동충하초와 눈꽃동충하초의 배양특성 및 항균활성)

  • Lee, Ki-Man;Hong, In-Pyo;Nam, Sung-Hee;Sung, Gyoo-Byung;Bae, Yoon-Hwan
    • Korean journal of applied entomology
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    • v.47 no.4
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    • pp.479-486
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    • 2008
  • The cultural characteristics and antibacterial activities of Cordyceps militaris and Paecilomyces tenuipes were compared. The mycelial growth was the highest on MCM (Mushroom Complete Medium) for C. militaris and on YMA (Yeast Malt Agar) for P. tenuipes. But the mycelial density on MMM (Mushroom Minimal Medium) was lower than other on media. The optimum mycelial growth was observed at $25^{\circ}C$. C. militaris was low mycelial growth when it was transferred over 5 times generation. The carbon source for the optimum mycelial growth was fructose of monosaccharide, maltose of disaccharide and dextrin of polysaccharide. The calcium nitrate of organonitrogen was found the best mycelial growth on C. militaris, while the sodium nitrate observed to be well for mycelial growth on P. tenuipes. The ammonium tartrate was observed to be the best among the inorganonitrogen used for mycelial growth. Antibacterial activities were found out just C. militaris against Bacillus cereus of Gram (+).

Plan for Countermeasure and Prospect of Demand and Supply about Seamen in Korea (우리나라 어선원 수급 전망과 대응방안)

  • Lee, Kwang-Nam;Cheong, Jin-Ho
    • The Journal of Fisheries Business Administration
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    • v.44 no.3
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    • pp.47-60
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    • 2013
  • A study on the seamen's demand - supply has been focused on early 1990s, and never studied yet since 2007. Because previous studies related to the seamen are very different in conditions the environments around the fishery recently. It is serious problem that the number of domestic semen are decreasing continually. To depend on foreign seamen because of lacking of domestic seamen should be linked closely with the fishing industry environment in our country. Therefore, this study is limited to domestic seamen. Lacking recent domestic seamen, it is the most likely to rise imbalance between demand - supply in the future. Through medium and long term its analysis based on data, we are willing to discover various political subjects for seamen's welfare, the training of manpower, education, etc. To solve these issues, it can be made institutional changes as follows ; First, it is necessary policy-making related the deteriorating employment situation based on its analysis of medium and long term for seamen. Second, it is necessary to overcome a dual system for its management for seamen. Third, it is necessary to improve problems of statistics data for fishery and build a statistical system for seamen. Also, it will be improved a insurance system under 5 ton and strengthen the function for Korean seamen welfare and employment center and must be urgently countermeasures for the minimal unemployment rate through a content development and management of home page. Finally, it will be supplied reliably seamen to improve a practice system for apprentices according to the international regulations(STCW-F) for fishery students.

Effects of Increased NADPH Concentration by Metabolic Engineering of the Pentose Phosphate Pathway on Antibiotic Production and Sporulation in Streptomyces lividans TK24

  • Jin, Xue-Mei;Chang, Yong-Keun;Lee, Jae Hag;Hong, Soon-Kwang
    • Journal of Microbiology and Biotechnology
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    • v.27 no.10
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    • pp.1867-1876
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    • 2017
  • Most of the biosynthetic pathways for secondary metabolites are influenced by carbon metabolism and supply of cytosolic NADPH. We engineered carbon distribution to the pentose phosphate pathway (PPP) and redesigned the host to produce high levels of NADPH and primary intermediates from the PPP. The main enzymes producing NADPH in the PPP, glucose 6-phosphate dehydrogenase (encoded by zwf1 and zwf2) and 6-phosphogluconate dehydrogenase (encoded by zwf3), were overexpressed with opc encoding a positive allosteric effector essential for Zwf activity in various combinations in Streptomyces lividans TK24. Most S. lividans transformants showed better cell growth and higher concentration of cytosolic NADPH than those of the control, and S. lividans TK24/pWHM3-Z23O2 containing zwf2+zwf3+opc2 showed the highest NADPH concentration but poor sporulation in R2YE medium. S. lividans TK24/pWHM3-Z23O2 in minimal medium showed the maximum growth (6.2 mg/ml) at day 4. Thereafter, a gradual decrease of biomass and a sharp increase of cytosolic NADPH and sedoheptulose 7-phosphate between days 2 and 4 and between days 1 and 3, respectively, were observed. Moreover, S. lividans TK24/pWHM3-Z23O2 produced 0.9 times less actinorhodin but 1.8 times more undecylprodigiosin than the control. These results suggested that the increased NADPH concentration and various intermediates from the PPP specifically triggered undecylprodigiosin biosynthesis that required many precursors and NADPH-dependent reduction reaction. This study is the first report on bespoke metabolic engineering of PPP routes especially suitable for producing secondary metabolites that need diverse primary precursors and NADPH, which is useful information for metabolic engineering in Streptomyces.

Evaluation of the Luminous Intensity Distribution of Stop Lamp for Agricultural Tractors (농업용 트랙터 제동등 광도 성능 평가)

  • Kim, Hyung-Kweon;Kim, Yu-Yong;Shin, Seoung-Yeoup;Kim, Byoung-Gap;Lee, Ho-Sang
    • Journal of Biosystems Engineering
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    • v.36 no.6
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    • pp.416-421
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    • 2011
  • This study was initiated to prevent rear-end collision which causes the most frequent human damage by agricultural tractors. Therefore, this study was aimed to evaluate the luminous performance of a stop lamp for an agricultural tractor which could reduce rear-end collision. The performance test of luminous intensity distribution was conducted for a total of 12 tractor models classified by size (above 44 kW for large, 30~43 kW for medium, and below 29 kW for small) and production area (domestic, foreign manufacture) in accordance with Regulation No.7 of UNECE Vehicle Regulations-1958 Agreement considering ${\pm}$20 standard deviation. Three out of 5 large, 3 out of 4 medium, and all small tractors were resulted as inadequate for the test criterion. This study found that the luminous performance results were better as the size of agricultural tractors became bigger. One out of 4 foreign and 2 out of 8 domestic manufacturers were suitable for the test criterion, and overall no difference from the luminosity test results was observed between domestic and foreign manufacturers. In addition, the measuring points, HV, 5L, 5R, 5U and 5D with the minimal luminosity revealed flaws (80% of the proportion of faulty products). Therefore, it is urgent to improve the quality for measuring points (HV, 5U, 5L, 5R, 5D) of stop lamps to apply Regulation No. 7.

Physiological Activity of Alliin and Ethanol Extract from Korean Garlic (Allium Sativum, L.) (한국산 마늘로 부터 분리한 Alliin과 에탄올 추출물의 In Vitro계 생리 활성)

  • Lim, Seung-Woo;Kim, Tae-Hyeo
    • Korean Journal of Food Science and Technology
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    • v.29 no.2
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    • pp.348-354
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    • 1997
  • This study was designed to evaluate the effects of garlic extracts of various concentrations on the growth of various pathogens and human colon cancer cell lines in vitro. For antibacterial effects against microorganisms, minimal inhibition concentrations (MIC) of alliin were from 5,000 to 20,000 ppm. MIC of ethanol extract were from 1,250 to 10,000 ppm. For cytotoxic effect of alliin and ethanol extract against human colon cancer cell lines (HCT-15), the growth rates of the cancer cells in medium containing alliin and ethanol extract were inhibited gradually to a significant degree in proportion to the increase of these concentration. Morphology of HCT-15 cells in medium containing alliin and ethanol extract were seen to be shrinked and fragmented. The results show that the causes of the antibacterial and cytotoxic effect against a wide range are thermostable substances isolated by the ethanol.

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