• Title/Summary/Keyword: Agar 농도

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Optimization of Genetic Transformation Conditions for Korean Soybean Cultivars (국내 콩(Glycine max) 품종 형질전환 초기조건 확립)

  • Lee Ki-Jung;Seo Jen-Kyung;Lee Hye-Young;Jeon Eun-Hee;Shin Sang-Hyun;Lee Jai-Heon;Kim Doh-Hoon;Ko Jong-Min;Hahn Won Young;Baek In-Youl;Oh Boung-Jun;Chung Young-Soo
    • Journal of Life Science
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    • v.16 no.2 s.75
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    • pp.289-296
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    • 2006
  • In order to establish highly efficient gene transfer condition at early stage of soybean transformation, various experiments were performed and compared their efficiencies by transient GUS analysis; those conditions are genotype determination of Korean soybean cultivars for amenability to Agro-infection, appropriate agar and selective agent concentration, orientation of explant placement, hormone pre-culture, and liquid selection condition. In the genotype screen of Korean soybean varieties, 14 amenable genotypes were selected. For efficient Agrobacterium washing, cefotaxime was chosen and hygromycin at the concentration of 10 and 15 ppm was used as selection agent in the media. Agar concentration was slightly better in 0.6% and 0.8% for both shoot and callus formation, and explant placement with adaxial side down showed high frequency of GUS expression. For wounding treatment, oriental needle was efficient than scalpel for shoot formation and gene transfer. To increase the frequency of gene transfer, hormone pre-treatment was applied. BA at the concentration of 5 and 10 ppm resulted in better survival at the late stage of selection in shoot elongation media. Selection in liquid media after hormone pre-treatment seemed to be effective to remove the escaped non-transformants at early stage of procedure. Considering the results obtained, Eunhakong could be the first choice as a material for soybean transformation among Korean soybean genotypes.

Residual Activity and Effect of Soil Applied Herbicides on Succeeding Crops in Vegetable Field 3. Growth Inhibition of Test Plant According to Concentration Level of Herbicides in Sterile Agar Medium (주요채소용제초제(主要菜蔬用除草劑)의 토양중(土壤中)에서의 잔효(殘效)과 후작물(後作物)에 미치는 영향(影響) 제(第)3보(報) 무균한천배지(無菌寒天培地)에 있어서 제초제(除草劑)의 농도수준(濃度水準)에 따른 검정식물(檢定植物)의 생육저해(生育沮害))

  • Ryang, H.S.;Moon, Y.H.;Choi, E.S.;Jang, M.S.;Lee, J.H.;Chang, Y.N.
    • Korean Journal of Weed Science
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    • v.11 no.2
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    • pp.117-121
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    • 1991
  • This experiment was conducted to identify the relationships of concentration with phytotoxic rate of dinitroamide and acid amide herbicides which have showed the longest residual period in soil. Four herbicides treated showed greater inhibition on roots than shoots, greater inhibition by herbicides was obtained for Italian ryegrass than for Radish. Nitralin, pendimethalin, and ethalfluralin at 0.01ppm gave about 20% inhibition of Italian ryegrass roots, whereas about 47% inhibition was obtained with napropamide. Fifty percent inhibition($I_{50}$) of roots and shoots of Italian ryegrass was 0.036 and 0.132ppm for nitralin. 0.063 and 0.097ppm for pendimethalin. 0.042 and 0.092ppm for ethalfluralin. 0.027 and 0.071ppm for napropamide respectively. On the other hand, $I_{50}$ of roots and shoots of Radish was 1.028 and over 10ppm for nitralin. 1.925 and 4.885ppm for pendimethalin, 2.669 and over 10ppm for ethalfluralin, and 0.515 and over 10ppm for napropamide respectively. There was positive correlation between the concentration of herbicides and growth inhibition of Italian ryegrass and radish.

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Evaluation of Dry Rehydratable Film Method for Detection of Coliform Bacteria and Escherichia coli (대장균군과 대장균의 검출을 위한 건조필름 방법의 평가)

  • Park, Heon-Kuk
    • The Korean Journal of Food And Nutrition
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    • v.22 no.4
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    • pp.696-700
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    • 2009
  • The adaptability of dry rehydratable film for the qualitative evaluation of coliform bacteria and Escherichia coli was tested. In general, culture methods that employ lactose broth or desoxycholate lactose agar are used for qualitative tests of coliform bacteria. Culture using lactose broth showed a high detection yield and low selectivity when compared to the dry rehydratable film. However, culture methods that employ lactose broth required a long time(48 hrs) for qualitative tests of coliform bacteria and complicated procedures were required to prepare the medium. The detection of coliforms using desoxycholate lactose agar had a slightly higher selectivity than the dry rehydratable film method, but this difference was not statistically significant. However, the preparation of the desoxycholate lactose agar was complicated. EC broth for the detection of E. coli showed the highest detection yield and lowest selectivity; however, this method required complicated procedures for preparation of the medium as well. Overall, the dry rehydratable film had a slightly lower detection yield than the other methods. The detection yield of dry rehydratable film method was over 37.1% at a concentration of 1 cfu/$m{\ell}$. Additionally, the dry rehydratable film method showed high selectivity and did not require preparation. However, because the selectivity of the dry rehydratable film was high, it took a long time(36 hrs) to detect E. coli. Overall, these findings indicate that dry rehydratable film can be used for qualitative detection of coliforms and E. coli.

Effect of green tea on the survival of Escherichia coli O157:H7 and Salmonella typhimurium in mayonnaise (마요네즈에 첨가한 녹차가 Escherichia coli O157:H7과 Salmonella typhimurium의 생존에 미치는 영향)

  • 박찬성;박금순
    • Korean journal of food and cookery science
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    • v.18 no.1
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    • pp.57-62
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    • 2002
  • The purpose of this study was to investigate the antibacterial activity of green tea in mayonnaise against pathogenic bacteria (Escherichia coli O157:H7 and Salmonella typhimurium). Mayonnaise was prepared with salad oil, egg yolk, sugar, salt and vinegar, and green tea powder was added to the mayonnaise at 0.1, 0.3 and 0.5% for the experiment. Then, the mayonnaise samples with various levels of green tea were inoculated with about 10$\^$6/ cells/g of E. coli and S. typhimurium per 1 gram of mayonnaise and stored at 5$^{\circ}C$, 15$^{\circ}C$ and 25$^{\circ}C$ for 3∼9 days. Antibacterial activity of green tea was tested by the colony counting method for E. coli on violet red bile agar(VRBA) and S. typhimurium on xylose lysine desoxycholate agar(XLDA). The D-values of E. coli controls were 2.89, 2.73 and 2.13 days while those of S. typhimurium controls were 0.58, 0.53 and 0.52 days at 5$^{\circ}C$, 15$^{\circ}C$ and 25$^{\circ}C$, respectively Inhibitory effect of green tea in mayonnaise on the survival of E. coli and S. typhimurium was increased with increasing concentration of green tea and/or increasing storage temperature. The most effective antibacterial activity of green tea was shown against E. coli in mayonnaise during storage at 25$^{\circ}C$.

Development of L-Lysine Producing Strains from Cellulosic Substrate by the Intergeneric Protoplast Fusion- Conditions for Formation and Regeneration of Protoplast - (속간 원형질체 융합에 의한 섬유질 기질로부터 L-lysine 생산균주 개발 -원형질체의 형성 및 재생 -)

  • 성낙계;정덕화;이무영;정영철
    • Microbiology and Biotechnology Letters
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    • v.16 no.2
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    • pp.150-155
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    • 1988
  • In order to produce L-lysine from cellulosic substrates by the intergeneric protoplast fusion between cellulolytic bacteria, Cellulomonas flavigena KFCC31221 and amino acid producing bacteria, Brevibacterium flavum ATCC14067, Corynebacteriurn glutamicum ATCC13032, conditions for protoplast formation and regeneration of these strains were investigated. After the strains were mutated with 500$\mu\textrm{g}$/$m\ell$ N-methyl-N'-nitro N-nitrosoguanidine for 30 min and the mutants were enriched by treating 300$\mu\textrm{g}$/$m\ell$ penicillin-G for 2 hrs, B. flavum Hse- Str$^{r}$ , C. glutamicum Met$^{-}$Thr$^{-}$ Rif$^{r}$ and Cellulomonas flavigena Thr$^{-}$Val$^{-}$Kan$^{r}$ were isolated. The rate of protoplast formation ranged from 95 to 98% when strains were treated at the concentration of 500$\mu\textrm{g}$/$m\ell$ of lysozyme, pH 6.5, 33$^{\circ}C$, for 6 hrs. in Tris- malate buffer supplemented with 0.4M sucrose as osmotic stabilizer. Approximately 30-33% protoplast was regenerated on the regeneration complete medium(RCM) containing 1.5% agar and 0.5M sodium succinate overlaid with the same medium except 0.7% agar.

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Isolation of an Agarolytic Bacteria, Cellvibrio mixtus SC-22 and The Enzymatic Properties (한천분해세균 Cellvibrio mixtus SC-22의 분리 및 효소적 특성)

  • Cha, Jeong-Ah;Kim, Yoo-Jin;Seo, Yung-Bum;Yoon, Min-Ho
    • Journal of Applied Biological Chemistry
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    • v.52 no.4
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    • pp.157-162
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    • 2009
  • An agar-liquefying bacteria (SC-22), which produces a diffusible agarase that caused agar softening around the colony was isolated from Daecheong lake in Korea. Chemotaxanomic and phylogenetic analyses based on 16S rRNA gene sequences revealed the strain was classified as Cellvibrio mixtus SC-22. The isolate SC-22 showed maximal extracellular agarase activity with 58.5 U/mL after 48 h cultivation in the presence of 0.2% agar. It was observed that the isolate produced two kinds of extracellular and three kinds of intracellular isoenzymes. The major agarase was purified from the culture filtrate of agarolytic bacteria by ammonium sulfate precipitation, anion exchange and gel filtration column chromatographic methods. The molecular mass of the purified enzyme was estimated to be 25 kDa by SDS-PAGE. The optimum pH and temperature of the purified enzyme were pH 7.0 and $50^{\circ}C$, respectively. The agarase activity was activated by $Fe^{2+}$, $Na^+$ and $Ca^{2+}$ ions while it was inhibited by $Hg^{2+}$, $Mn^{2+}$ and $Cu^{2+}$ at 1 mM concentration. The predominant hydrolysis product of agarose by the enzyme was galactose and disaccharide on TLC, indicating the cleavage of $\beta$-1,4 linkage in a random manner. The enzyme showed high substrate specificity for only agar and agarose among various polysaccharides.

An Optimal Standardized in vitro Bioassay to Evaluate Susceptibility of Green Peach Aphid, Myzus persicae (Sulzer)(Insecta: Hemoptera: Aphididae), to Aphicides (복숭아혹진딧물, Myzus persicae (Sulzer)(Insecta: Hemoptera: Aphididae), 살진딧물 최적 in vitro 살충력 검정 방법 확립)

  • Ka Hee Cho;Hyo Jung Kim;Young Cheol Kim
    • Korean journal of applied entomology
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    • v.62 no.3
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    • pp.139-147
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    • 2023
  • Leaf-spray in vitro bioassays appraise new aphicidal formulations for managing deleterious plant-feeding aphids. The formulation may utilize alternative and integrated strategies. However, leaf spraying even under controlled conditions may affect aphid reproduction and mortality. This study examines leaf spray applications for optimum and reproducible aphicidal results using tobacco leaves overlaid on cotton fabric or water agar surfaces. Infestation of the undersides of tobacco leaves with nymphs of green peach aphids was used in the assays. Spray distance and volume were optimized using water-sensitive paper to ascertain the best surface coverage. Overlays of the leaves on water agar caused less mortality and greater reproduction than the use of cotton fabric. The relative humidity of the insect-rearing chambers changed with the watering regime for the insect - rearing chambers with cotton fabric; 60% relative humidity was optimal. Relative humidity was not affected by the concentration of agar in the water agar chambers. Applications of the chemical aphicidal standard, Sulfoxaflor, under the optimized conditions exhibited similar times for lethality although the rate was faster with leaves on the cotton fabric than on water agar. These studies establish reproducible and sensitive techniques for assessing the lethality and effects on reproduction of potential aphicidal products.

Effect of Growth Medium and Extract Sterilization on a Seedling Bioassay of Alfalfa Autotoxicity (생장배지 및 추출액의 멸균이 알팔파 Autotoxicity 의 유묘 생물검정에 미치는 영향)

  • Chon, Sang-Uk;Nelson, C.Jerry;Han, Sung-Uk;Guh, Ja-Ock
    • Korean Journal of Environmental Agriculture
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    • v.18 no.2
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    • pp.185-190
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    • 1999
  • Autotoxic substance(s) from alfalfa(Medicago sativa L.) plants reduces germination and growth of adjacent new alfalfa after alfalfa. The autotoxic chemical(s) in alfalfa are clearly unknown. Our objective was to improve the sensitivity of an alfalfa seedling bioassay for evaluating autotoxic leaf extracts. We determined critical extract concentrations that inhibit seed germination and seedling growth, compared two different culture media, and evaluated the effects of extract sterilization on the sensitivity of the assay, by using streptomycin and autoclaving method. An agar medium in petri plate gave better responses of germination and seedling growth to the extracts than using filter paper in the plate. On agar medium, the concentration of extract required to reach 50% inhibition of root length was 2.7 g $kg^{-1}$, and of germination and hypocotyl length were 3.8 and 9.9 g $kg^{-1}$, respectively. Leaf extracts with 100 ppm streptomycin stimulated germination significantly compared to Leaf extract alone but reduced root length of control by 43%. Root length was more sensitive to the autotoxin(s) than was germination or hypocotyl length. These results suggest that agar medium mixed with extract and sterilization by autoclaving could be improved the consistency and precision of bioassay, and that root length was the best parameter of autotoxic effect of alfalfa leaf extract.

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Effects of Media and Temperatures on Micro Stem Cutting of Dendrobium nobile 'Hamana Lake Dream' × 'No. 55' in Vitro (Dendrobium nobile 'Hamana Lake Dream' × 'No.55'의 기내 줄기삽목에 미치는 배지의 종류와 온도의 영향)

  • Yoon, Jin-Young;Nam, Yu-Kyeong;Lee, Jong-Suk;Kim, Hyun-Jin
    • Journal of agriculture & life science
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    • v.44 no.3
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    • pp.23-30
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    • 2010
  • The optimal growth condition of in vitro stem cutting of Dendrobium nobile 'Hamana Lake Dream' ${\times}$ 'No. 55' was investigated. Among various media and their concentrations, MS media had better effect on the growth of micro stem cutting than Hyponex media in all concentration levels except stem length. The activated charcoal concentration in MS media showed different effects on number of stem and root, leaf length, and fresh weight: the most effective in the range of 0.1 to 1.0 g/L and barely effective above 2.0 g/L. Addition of agar 5 g/L, sucrose at 40 g/L, and peptone at 1 g/L to MS media increased significantly stem length, leaf width, and fresh weight, internode length and number of roots, and the number of stem and leaves. On the other hand, addition of gelite with any concentration had no effect on the growth of micro stem cutting compare to that of control. The optimal temperature for growth of micro stem cutting was $28^{\circ}C$. Under the same temperature, MS medium was better than Hyponex medium for the growth of stem. In addition, sucrose at 40 g/L was the most effective on growth at $28^{\circ}C$.

Efficiency of PetrifilmTM Staph Express Count Plate for the Enumeration of Staphylococcus aureus in Meat, Fishery Product, and Korean Traditional Foods (축·수산·전통식품 중 황색포도상 구균의 정량적 분석을 위한 PetrifilmTM Staph Express Count Plate의 성능 평가)

  • Yoo, Yoonjeong;Choi, Yuna;Choi, Seungho;Bang, Hyunjo;Yoon, Yohan;Ha, Jimyeong;Lee, Soomin
    • Journal of Food Hygiene and Safety
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    • v.34 no.3
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    • pp.309-315
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    • 2019
  • To enumerate Staphylococcus aureus in food, Baird-Parker Agar (BPA) is usually used in the conventional method, However it requires time and space for the preparation and plating, and incubation. Thus, use of the $3M^{TM}$ $Petrifilm^{TM}$ Staph Express Count Plate (STX Petrifilm) might be appropriate to solve these challenging problems. The purpose of this study was to compare the efficiency of STX Petrifilm with BPA for enumeration of S. aureus in various foods. A mixture of S. aureus strains ATCC29213, ATCC25923, and ATCC13565 was inoculated on marinated pork chop, beef (chuck tender), dried filefish, semi-dried squid, rice cake, and Japchae (stir-fried glass noodles) at 2, 3, 5, and 7 Log CFU/g. S. aureus cell counts were enumerated by spread-plating on STX Petrifilm and BPA after 0 and 24 hours at $4^{\circ}C$ (marinated pork chop, beef, semi-dried squid, and stir-fried glass noodles) and $25^{\circ}C$ (dried filefish and rice cake). Recovery of STX Petrifilm for S. aureus from various food samples was compared with BPA, and the results showed that there were no significant differences between two selective media in all cases. The results indicated that STX Petrifilm had enough efficiency to recover S. aureus from various foods as well as saving time and space.