• Title/Summary/Keyword: 계대 배양

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In vitro micropropagation of radish (Raphanus sativus L.) using callus induction and plant regeneration (캘러스 유기와 식물체 재분화를 이용한 무의 기내 대량증식)

  • You Kyoung Kim;Sug Youn Mo;Su Bin Choi;Han Yong Park
    • Journal of Plant Biotechnology
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    • v.50
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    • pp.155-162
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    • 2023
  • Radish (Raphanus sativus L.), a root vegetable grown worldwide, is consumed in several ways. In the cross between parental lines to produce F1 seeds of radish, the problem of low purity may arise because of pollen contamination. Therefore, we aimed to establish conditions for callus induction and regeneration so that in vitro cultured plants could be used for the propagation of stock seeds. The most effective hormone combination containing various concentrations of 2,4-D, TDZ, and kinetin was selected for callus induction using radish hypocotyl, and the induced calli were transferred to two types of hormone media to investigate the optimal conditions for shoot regeneration of the callus. The combination of 1 mg/L 2,4-D + 0.05 mg/L kin was the most effective for callus induction of RA2 and RA10, 1 mg/L 2,4-D + 0.1 mg/L kin + 0.025 mg/L TDZ of RA4, and 1 mg/L 2,4-D + 0.2 mg/L kin of RA30. Shoot regeneration of the RA4 callus occurred in both shoot regeneration media, but the frequency was much higher in the 5H+1B medium (1 mg/L NAA + 0.1 mg/L 2,4-D + 1 mg/L IPA + 0.02 mg/L GA3 + 2 mg/L zeatin + 1 mg/L BA). For the in vitro micropropagation of radish, the conditions selected in this study can assist in the propagation and maintenance of stock seeds to produce F1 seeds.

Bacterial Studies on the Subsidiary Materials of Fish Sausage (어육소시지 부원료에 대한 세균학적 연구)

  • 조갑숙;김성준;이응호
    • Microbiology and Biotechnology Letters
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    • v.8 no.3
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    • pp.155-166
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    • 1980
  • Studies have teen undertaken to investigate the degree of microbial contamination in the subsidiary materials which have been known as an important source of microorganisms associated with spoilage of fish sausage and fish paste products. Twenty hinds of food ingredients including starch, spices and condiments, 59 samples in total collected from commercial fish sausage processing plants and supermarket in the period of July to October 1979, were examined for standard plate count, coliform and fecal coliform, mold and yeast, thermoduric microorganisms, aerobic sporeformers (mesophilic and thermophilic), anaerobic sporeformers (mesophilic and thermophilic) and sulfide spoilage anaerobes. The results obtained are summarized as follows. 1. Among the food ingredients examined, corn starch, black pepper, hot pepper, onion, garlic, ginger, beef extract and frank marked high bacterial contamination with general and sporeforming microorganisms. And bacterial content of marked samples were generally higher than that of the samples from plants. 2. The high standard plate count caused by high content of these bacteria like thermoduric, mesophilic or thermophilic sporeforming aerobes. 3. Bacterial content of food ingredients such as black pepper and beef extract being used in plants, and black pepper, hot pepper, onion and garlic from the market were exceeded the bacterial standards being enforced in Japan and U. S. A. 4. Average standard plate count was in the range of 10$^4$to 10$^{5}$ /g for black pepper, wheat flour, onion and garlic collected from plants, and 10$^{5}$ to 10$^{7}$ /g for black pepper, hot pepper, onion and garlic from market. No plate count was observed in pepper essence and coloring material. 5. Coliform organism was detected in starch, black pepper, hot pepper, onion, garlic, ginger and gluten that showed high standard plate but no fecal coliform in the samples except black pepper and hot pepper. 6. Average mold and yeast count was 140 to 460/g for corn starch, wheat flour and black pepper from plants, and 10$^3$/g for black pepper and hot pepper from market. No count was observed in the other ingredients. 7. Sulfide spoilage sporeforming anaerobes boiled for 5 min. at 10$0^{\circ}C$ and incubated at 55$^{\circ}C$ was not detected in all the samples examined.

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The Study on Rough Colony Type Mutant of Pseudomoms mori(Boyer et Lambert) Stevens, caused Mulberry Bacterial Blight: Pathogenicity and General Characteristics (뽕나무 세균성위축병균 Pseudomonas mori (Boyer et Lambert) STEVENS의 Rough Colony Type Mutant에 관한 연구 ; 병원성 및 일반적 성질)

  • Yi Young Keun;Kim Jong Wan;Cho Yong Sup
    • Korean journal of applied entomology
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    • v.16 no.1 s.30
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    • pp.21-31
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    • 1977
  • The study has been carried out to compare the pathogenicity, physiological characteristics and genetic reliability between rough colony type strain and smooth colony type strains of Pseudomonas mori (Boyer et Lambert) Stevens which were isolated from diseased plant parts in 5 different areas throughout country. The results are summarized as follow. 1. The rough colony type strain showed more agressive reactions to tested host plant varieties than smooth colony type strains though there was no differences in the appearence of lesion types caused by both strains. 2. Both colony types were differentiated morphologically in that the rough colony type strain was having more than 200r long filamentous body without flagella where as the smooth colony type strains have short rods with one or several polar flagella. 3. The colony of smooth type strains was circular, entire, smooth and opaque, while the rough type strain shelved endulated, irregular margin, rough and wrinkled colony on nutrient agar media. 4. There were no differences between both colony types in the physiological and serological test. 5. Both of smooth and rough colony type strains showed genetic reliability through more than 100 succeeding cultures on the media, and were stable to various chemicals such as 1 to 3 percent of NaCl, 5 kinds of organic acid and 4 kinds of antibiotics.

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Growth Characteristics and Physiological Properties in Milk of Lactobacillus casei CU2604 Isolated from Adult Feces (성인으로부터 분리된 Lactobacillus casei CU2604의 우유배지에서의 생장 특성 및 생리적 특성)

  • Kim, Hee-Jin;Choi, Jae-Kyoung;Lee, Kyung-Min;Im, Jung-Hyun;Eom, Seok-Jin;Kim, Geun-Bae
    • Food Science of Animal Resources
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    • v.29 no.5
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    • pp.619-626
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    • 2009
  • As a trial for the development of a new starter culture for yogurt products, more than two hundred lactic acid bacteria strains were isolated from raw milk and healthy human feces. The strains that showed excellent growth and acid production ability in the 10% skim milk media were selected and identified as Lactobacillus casei through the API carbohydrate fermentation pattern and 16S rDNA sequence analysis. L. casei CU2604 was further investigated for its physiological characteristics as a starter culture compared with a commercial strain. The CU2604 strain showed good acid production and growth characteristics in milk, which were comparable to those of the L. casei Shirota strain. Despite the fact that both these strains displayed the same sugar fermenting pattern and PFGE band pattern, and had similar growth characteristic in milk, L. casei CU2604 exhibited different fatty acid composition in the cell wall, showed more tolerance to bile and to pH, and presented better growth inhibition activity against pathogenic bacteria. Based on these results, the L. casei CU2604 strain holds great promise for use as a novel and efficient starter culture in the production of yogurt. Additional studies on the probiotic characteristics of this strain are currently being conducted.

Somatic embryo induction and plant regeneration from cold-stored embryogenic callus of K. septemlobus (저온저장 음나무 배발생 캘러스로부터 체세포배 유도와 식물체 재생)

  • Lee, Na Nyum;Choi, Yong Eui;Moon, Heung Kyu
    • Journal of Plant Biotechnology
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    • v.42 no.4
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    • pp.388-395
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    • 2015
  • Somatic embryogenesis is as an excellent technology for potential use in plant mass production, germplasm conservation, or genetic engineering. We examined the effect of cold storage using 3 embryogenic callus lines with different levels of embryogenesis competence derived from immature zygotic embryo cultures of Kalopanax setemlobus. Somatic embryo induction, germination and plant conversion were evaluated after 1, 3 and 6 months storage at $4^{\circ}C$ in the dark. Most cold-stored embryogenic calli formed somatic embryos normally even after 6 months; however, the induction rate was gradually decreased by increasing the storage period. The most competent line tended to show a slight decline in somatic embryo induction rate, as compared with other lines after cold storage. In general, cold storage resulted in reduced somatic embryo germination and plant regeneration, although 93% somatic embryo germination and 91% plant conversion were achieved regardless of the storage period. Cold storage led to cell browning and degradation. Additionally, the cell structures were confirmed by the aceto-carmine and evans blue dye evaluation. Collectively, our results showed that embryogenic callus of K. septemlobus could be preserved at $4^{\circ}C$ without subculture for 6 months, and suggested the need for storage of relatively more competent embryogenic calli lines to support somatic embryo induction.

Development of Herbicide(BIALAPHOS) Tolerant Tobacco through Tissue Culture (제초제(除草劑) BIALAPHOS에 대(對)한 연초(煙草)의 내성(耐性) 증대(增大))

  • Bae, Y.Z.;Kim, K.U.
    • Korean Journal of Weed Science
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    • v.8 no.2
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    • pp.182-186
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    • 1988
  • This study was conducted to level up the tolerance of tobacco plant against bialaphos herbicide through tissue culture. The relatively good shoot regeneration from the subcultured calli treated with bialaphos at 0.5 ppm was observed in old the tobacco varieties tested such as NC 82, BY 4 and KA 101. However, at the treatment of bialaphos 1.0 ppm, shoot regeneration was only made in KA 101 variety, showing better regeneration than that of untreated one, When these shoots were transfered to the medium containing of bialaphos 10.0 ppm, the percentage of living shoots (i.e. tolerant plant) was very low, showing 2.43% in NC 82, 2.76% in KA 101 and 0.78% BY 4. Calli were induced and multiplied from leaf petiole of the above tolerant plants even under 2.5ppm of bialaphos, showing an average of 9% in NC 82 and 16% in KA 101 as compared with the untreated control. No calli were induced from tolerant plants as bialaphos concentration increased up to 5.0 ppm. Direct shooting from leaves of the above tolerant plants, that is selected at 10.0ppm of bialaphos treatment, was observed even under 10.0ppm of bialaphos treatment both in NC 82 and in KA 101 varieties, indicating that tolerance of tobacco plants against bialaphos can be greatly increased.

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Transgenic Sweetpotato (Ipomoea batatas) Expressing Spike Gene of Porcine Epidemic Diarrhea Virus (돼지 유행성 설사병 바이러스의 스파이크 유전자 발현 형질전환 고구마)

  • Yang Kyoung-Sil;Lim Soon;Kwon Suk-Yoon;Kwak Sang-Soo;Kim Hyun-Soo;Lee Haeng-Soon
    • Journal of Plant Biotechnology
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    • v.32 no.4
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    • pp.263-268
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    • 2005
  • Porcine epidemic diarrhea virus (PEDV) causes acute enteritis in pigs of all ages and is often fatal for neonates. In order to develop sweetpotato plants expressing PEDV antigen, we constructed the vector expressing spike gene of PEDV under the control of sweetpotato sporamin promoter or constitutive CaMV 35S promoter. The spike protein region of PEDV was synthesized by PCR and linked to each promoter, Transgenic sweetpotato [Ipomoea batatas (L.) Lam. cv. Yulmi] plants were developed from embryogenic calli following Agrobacterium tumefaciens-mediated transformation. The co-cultured embryogenic calli transferred to selective MS medium containing 1 mg/L 2,4-D, 100 mg/L kanamycin, and 400 mg/L claforan. These embryogenic calli were subcultured to the same selection medium at 3 weeks interval. Kanamycin-resistant calli transferred to hormone-free MS medium with kanamycin gave rise to somatic embryos and then converted into plantlets in the same medium. Southern blot analysis confirmed that the spike gene of PEDV was inserted into the genome of the sweetpotato plants. RT-PCR revealed that the spike gene of PEDV was highly expressed in transgenic sweetpotato plants.

Differential Potential of Stem Cells Following Their Origin - Subacromial Bursa, Bone Marrow, Umbilical Cord Blood - (줄기세포의 분화능의 기원에 따른 비교 - 견봉하 점액낭, 골수, 탯줄 혈액 -)

  • Sim, Sung Woo;Moon, Young Lae;Kang, Jung Hun
    • Clinics in Shoulder and Elbow
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    • v.15 no.2
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    • pp.65-72
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    • 2012
  • Purpose: To evaluate the differentiation potential of stem cells and their immunophenotype from 3 different sources. Methods: Our study involved three stem cell sources-subacromial bursal tissue, bone marrow, and umbilical cord blood. We obtained the subacromial bursal tissue and bone marrow from the patients undergoing shoulder surgery. After collecting the sample, we applied specific induction media for neurogenic, adipogenic and osteogenic differentiation. Also, flow-cytometry analysis was done to reveal the cell surface antigens. Results: We obtained 100% (8 cases) neural and adipogenic differentiation, but 62.5% (5 of 8 cases) osseous differentiation among the subacromial bursal tissue group. Bone marrow derived cells showed 100% neural (6 cases) and adipogenic (5 cases) differentiation, but 80% (4 of 5 cases) osseous differentiation. Umbilical cord blood derived cells revealed 97% (65 of 67 cases) neural, 53.7% (29 of 54 cases) adipogenic and 68.4% (39 of 57 cases) osseous differentiation. Immunophenotype analysis revealed that surface markers of bone marrow, subacromial bursal cell and umbilical cord blood derived mesenchymal stem cells are different from each other. Conclusions: Mesenchymal stem cells are potential agents in regenerative medicine and are characterized by expression of surface markers and by their differentiation potential. Our study with stem cells from subacromial bursal tissue, bone marrow and umbilical cord discovered that each stem cell has unique differentiation potential and function based on its origin. Various stem cells show multi-lineage differentiations in vitro which can be correlated to in vivo conditions.

The Use of the Pathogen-specific Bacteriophage BCP8-2 to Develop a Rice Straw-derived Bacillus cereus-free Starter Culture (단일 박테리오파지를 이용한 볏짚 유래 Bacillus cereus free 스타터 컬쳐의 개발)

  • Bandara, Nadeeka;Chung, Seo-Jin;Jeong, Do-Youn;Kim, Kwang-Pyo
    • Korean Journal of Food Science and Technology
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    • v.46 no.1
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    • pp.115-120
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    • 2014
  • The purpose of this study was to develop a rice straw-derived Bacillus cereus (B. cereus)-free starter culture for traditional soybean fermented products using a B. cereus-specific bacteriophage, BCP8-2. To determine the optimal medium that supports the growth of rice straw-derived microorganisms and BCP8-2 activity, 5 different culture media were tested. The 5% ground bean (GB) medium was selected for further study. No B. cereus was detected in the BCP8-2-treated rice straw in GB medium, whereas B. cereus at a level of $10^7$ CFU/mL was recovered in the no-phage control. The total bacterial count reached approximately $10^9$ CFU/mL regardless of phage addition. When the 16S rRNA sequence-based microbial community was monitored using denaturing gradient gel electrophoresis (DGGE) and pyrosequencing, a similar microbial community was observed in the phage-treated and control samples. In conclusion, we demonstrate that phage can be used to prepare a rice straw-derived B. cereus-free starter culture with minimal effect on natural microflora.

Breeding of Yeast Strain with Starch Utilizing and Alcohol Fermenting Ability by Protoplast Fusion (전분분해활성과 알코올 발효능을 보유한 효모의 육종)

  • Ju, Min-No;Hong, Sung-Wook;Kim, Kwan-Tae;Yum, Sung-Kwan;Kim, Gye-Won;Chung, Kun-Sub
    • Microbiology and Biotechnology Letters
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    • v.36 no.2
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    • pp.158-164
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    • 2008
  • The fusants which contain starch utilizing ability and alcohol fermenting ability were developed by protoplast fusion of Saccharomyces cerevisiae KOY-1 and Saccharomyces diastaticus KCTC 1804. Sacharomyces cerevisiae KH-12 was obtained by haploid induction from Saccharomyces cerevisiae KOY-1. The auxotropic mutants of yeast were obtained by using an ethylmethane sulfonate (EMS). The frequency of protoplast formation in Saccharomyces cerevisiae KOY-1 $(Met^-)$ and Saccharomyces diastaticus KCTC 1804 $(Trp^-)$ were 90.5% and 97.7%, respectively. The frequency of fusant formation was $1.79{\times}10^{-4 }$ for the regenerated protoplast and the 1,000 fusants were obtained. Fusant FA 776 was selected as a potential yeast which contain an alcohol fermenting ability in the starch medium. The genetic stability was 4.64% for 10 passages of generation. Fusant FA 776 produced 13mg/ml of alcohol in 24% starch medium and showed 1.86-fold higher alcohol fermenting ability than Saccharomyces diastaticus KCTC 1804.