• Title/Summary/Keyword: Sterile medium

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Cultivation of Nostoc flagelliforme on Solid Medium

  • Su Jianyu;Jia Shiru;Qiao Changsheng;Kim Jung-Gyu;Hong Wan-Hae;Cho Ki-An;Choi DuBok
    • Korean Journal of Environmental Biology
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    • v.23 no.2 s.58
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    • pp.135-140
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    • 2005
  • In order to construct an artificial cultivation of Nostoc flagelliforme on solid medium, we attempted to assess the viability of approaches, which utilized either BG-11 agar or sand medium using both sterile and non-sterile algal segments. In the trial in which the BG -11 agar medium was inoculated with the non-sterile algal segments, the algae exhibited the rapid growth in the initial 4 days of cultivation. However, after 4 days of cultivation, the growth rate of the algae slowed, and the algal growth was completely stopped by 7 days of cultivation. When the BG -11 medium was inoculated with the sterile algal segments, the algae exhibited the rapid growth for a longer period of 8 days, reaching a length of 24.9 mm. The growth rate during this period was measured to be $24.5\%$. After the 8 days of cultivation, the algal growth rate began to slow and had almost stopped by the 13 days of cultivation. On the other hand, when the sterile algal segments were inoculated onto a sand plate, the algal segments decomposed, reaching total decomposition after 11 days of cultivation. By way of contrast, the desiccation treatment samples continued to grow for 14 days of cultivation. After 14 days of cultivation, the algae achieved a length of 26.1 mm, with a growth rate of $30.6\%$. Our results indicate that periodic desiccation may constitute an effective strategy for the prevention of algal decomposition.

Methods of temperature measurement of medium in bottle during sterilization (병재배 배지살균 시 배지내부의 실시간 온도측정 방법)

  • Cheong, Jong-Chun;Jhune, Chang-Sung;Lee, Chan-Jung;Oh, Jin-A;Shin, Pyeong-Gyun
    • Journal of Mushroom
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    • v.9 no.2
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    • pp.74-79
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    • 2011
  • This study was carried out to develop a method related to measuring the temperature of sterile medium in bottle cultivation. When the medium is sterilized, the device is able to be inserted inside of the medium and the temperature can be directly measured in real time although high temperature and pressure are detected in the sterilization. This device can be able to measure the sterilization temperature in short intervals inside of autoclave and medium used in bottle cultivation. As the method were applied to the field cultivation of mushroom, we could produce mushroom in consistent manner through the optimum sterilization of the medium.

A novel method for high-frequency transgenic shoot regeneration via Agrobacterium tumefaciens in flax (Linum usitatissimum L.)

  • Beyaz, Ramazan;Darcin, E. Selcen;Aycan, Murat;Kayan, Mustafa;Yildiz, Mustafa
    • Journal of Plant Biotechnology
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    • v.43 no.2
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    • pp.240-247
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    • 2016
  • In this study, routinely used transformation method, which includes transferring explants onto co-cultivation medium after inoculating them with bacterial solution for a while, was compared with 3 different inoculation methods. In every 3 methods, hypocotyl explants excised from 7-day-old sterile flax seedlings having cotyledon leaves and no root system dried under air flow in sterile cabin for 35 min were inoculated with different volumes of bacterial solution at different inoculation periods. GV2260 line of Agrobacterium tumefaciens having 'pBIN 19' plasmid containing npt II (neomycin phosphotransferase II) gene and GUS reporter gene was used in transformation studies. After inoculation, hypocotyl segments of seedlings (0.5 cm in length) - were excised and left to co-cultivation for 2 days. Then, explants were transferred to regeneration medium supplemented with different antibiotics. The presence of npt-II and GUS genes in transformants was confirmed by PCR and GUS analysis. The highest results in all characters examined in all cultivars were obtained from the 2 inoculation method in which hypocotyls excised from seedlings inoculated with $500{\mu}l$ of bacterial solution after drying in sterile cabin for 35 min were used.

Isolation and Drug Sensitivity of Gram-Negative Rods from Bovine Udder Infections (젖소 유방염(乳房炎)으로부터 Gram 음성간균(陰性桿菌)의 분리(分離) 및 약제감수성(藥劑感受性)에 관(關)한 연구(硏究))

  • Lee, Cha Soo;Park, Cheong Kyu
    • Korean Journal of Veterinary Research
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    • v.20 no.2
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    • pp.79-84
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    • 1980
  • One hundred and eighteen cultures of Gram-negative rods isolated from cases of clinical bovine mastitis during lactation were examined for distribution of specific types, and activity of several antimicrobial agents to the isolates was determined by two-fold tube dilution method employing sterile whole milk as fluid medium. Of the isolates, 59.2% were Escherichia coli. Most of the remaining isolates were Klebsiella pneumoniae and Enterobacter aerogenes. Most Gram-negative rods(89.8%) were isolated from acute local and chronic mastitis. The cases of peracute systemic form with a marked symptoms of toxemia were associated with Escherichia coli and Klebsiella pneumoniae. The minimal lethal concentrations (MLC) of gentamicin and oxolinic acid in sterile whole milk were 16-128 times higher than the MLC obtained in trypticase soy broth (TSB), while the MLC of ampicillin and tetracycline in milk increased 2-4 times compared with TSB. Of the drugs tested, gentamicin was the most active antibiotics with MLC of $100{\mu}g/ml$ in sterile whole milk against all of Gram-negative rods isolated from bovine udder infections.

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Interspecific Hybrid of Nicotiana trigonophyllo X N. tabacum through in vitro Culture of Fertilized Ovules (배주배양에 의한 Nicotiana trigonophylla와 N. tabacum의 종간잡종 육성)

  • 최상주;이승철
    • Journal of the Korean Society of Tobacco Science
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    • v.13 no.2
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    • pp.52-58
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    • 1991
  • Interspecific cross between Nicotiana trigonophylla and N. tabacum cv. BY4 is highly sterile because of abnormal ovule and embryo development. In vitro culture of excised N. trigonophylla ovules after polination by N. tabacum allows significant numbers of hybrid embryos to develop into mature plants. Total yield of seedlings and number of normal seedlings were produced following in vitro culture of individual fertilized ovules of N. trigonophylla X N. tabacum at four days post-pollination on B5 medium containing 6% sucrose. Hybrids were uniform in morphology and peroxidase isozyme composition and the majority were cytologically stable: flower characteristics were generally intermediate between those of the parents. All hybrids evaluated were self-sterile.

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Breeding Hybrid Rice with Good Quality and High Yield I. Breeding of Cytoplasmic-Genetic Male Sterile and Restorer Lines with Backgrounds of Korean Rice Varieties (양질 다수성 일대잡종 벼 육성 연구 I. 우리나라 품종 배경의 세포질-유전자적 웅성불임 및 임성회복 계통 육성)

  • Hak-Soo SUH
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.38 no.5
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    • pp.413-417
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    • 1993
  • This study was conducted to breed various cytoplasmic genetic male sterile (CGMS) and restorer lines with backgrounds of Korean japonica rice varieties. The CGMS line BT-CMS was crossed with the restorer line AR-3. The fertile $F_1 was emasculated and crossed with the Korean japonica rice varieties of the early maturing Sobaegbyeo, Odaebyeo, Gwanagbyeo and Daeseongbyeo, and of the medium maturing Hwajinbyeo, Paldal, Suwon 224 and Iri 386, and of the late maturing Nagdongbyeo, Palkweng, Hwacheongbyeo and Milyang 97. Each of the three way cross $F_{1S} was segregated into fertile and sterile individuals. The sterile individuals in each cross were discarded and the fertile individuals were emasculated and backcrossed with the Korean japonica rice varieties. The same process was applied from BC$_1$F$_1$ to BC$_4$F$_1$ generation. In the ${BC_4}{F_1}of each cross, the male sterile individual was crossed with the recurrent Korean japonica variety which was maintainer of male sterility. The male sterile lines of ${BC_6}{F_1}were named as Sobaegbyeo R, Odaebyeo R, Gwanagbyeo R, etc. The fertile individuals homozygous in pollen fertility were selected from the ${BC_4}{F_2}generation and named as Sobaegbyeo R, Odaebyeo R, Gwanagbyeo R, etc. Agronomic characteristics of the CGMS, restorer lines with backgrounds of Korean japonica, and the recurrent Korean japonica rice varieties grown in the field condition were compared. Culm length of the CGMS lines tended to be shorter than that of the recurrent parent, however no significant differences in heading date, panicle length and yield component were found among the CGMS, restorer and recurrent lines.

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Detection of Xanthomonas axonopodis pv. citri, the causal agent of bacterial canker on Unshiu orange fruits using bacteriophage in Korea.

  • Myung, Inn-Shik;Lee, Young-Hee
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.135.1-135
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    • 2003
  • A technique for detection of Xanthomonas axonopodis pv. citri, a causal bacterium of canker on Unshiu orange fruits, was developed using bacteriophage. Procedure for the detection was designed on the basis of the previous reports that one group(CPI) of X. axonopodis pv. citri bacteriophage and corresponding two Iysotypes distributed in Korea. First, fruit surface was washed with sterile distilled water and pellet was obtained from centrifugation. The pellet was resuspended in Wakimoto's potato semi-synthetic broth medium and divided equally into two parts. One part was heated in boiling water to kill bacterial cells. Bacteriophages(CP$_1$) were respectively added into two parts and 0.1 ml from each part was mixed with soft agar medium. After incubation for 18 hrs at 25$^{\circ}C$, the causal bacterium of canker was determined based on plaques formed on the medium. This procedure can be effectively used for detection of living bacterial pathogen on fruit surfaces of Unshiu orange.

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A METHOD OF MUCOSA CULTURE (구강점막의 배양에 관한 연구)

  • Choi, Byung-Ho;Yoo, Jae-Ha
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.17 no.4
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    • pp.331-336
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    • 1995
  • To use cultured mucosa as a graft of full thickness, our laboratory has been involved in the development of techniques to grow epidermis together with connective tissue. Human oral mucosa was obtained at dental surgery. Under sterile conditions the tissues were cut into explants of 0.1 $cm^2$ which were placed in the center of 24 well tissue culture dishes and incubated in a growth medium. The growth medium used for epithelial was MEM(Minimum Essential Medium) supplemented with 10% fetal calf serum, 0.5% dimethyl sulfoxide, glutamine (0.292 g/l), epidermal growth factor (40 ug/ml), cholera toxin (30 ng/ml), hydrocortisone (2 ug/ml), insulin (40 ug/ml) and transferin (5 ug/ml). The medium for stratification of epithelial cells was MEM supplemented with 10% fetal calf serum, 0.5% dimethyl sulfoxide and glutamine (0.292 g/l). The medium used for fibroblasts was MEM supplemented with 10% fetal calf serum. With the three types of media used alternatively, a mucosa composed of epidermis and connective tissue was obtained after 3 weeks of culture.

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Production of Artificial Seeds by Alginate-encapsulation of Rice Somatic Embryos (벼의 수화겔 인공종자 생산)

  • 정원중;민성란;송남희;유장렬
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.3
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    • pp.183-186
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    • 1994
  • Somatic embryos derived from cell suspension cultures of rice were singly alginate-encapsulated to be used as artificial seeds. When placed on half strength MS solid medium,73% of the encapsulated somatic embryos were capable of germination Encapsulation per se did not affect the germination frequency of embryos. When incubated by wrapping with moistured non-sterile filter paper, 60% of the encapsulated somatic embryos germinated. However encapsulated zygotic embryos without endosperm showed a high germination frequency regardless of the sterility of the incubation conditions. The results suggest that a greater susceptibility of somatic embryos to contaminants is attributed to lower germination frequency of encapsulated somatic embryos in non-sterile conditions.

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Production of Sexual Hybrids Nicotiana rustica X N. tobacum via in vitro Culture .of Fertilized ovules. (배주배양에 의한 Nicotiana rustica와 N. tabacum의 잡종식물 육성)

  • 최상주;홍병희
    • Journal of the Korean Society of Tobacco Science
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    • v.14 no.1
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    • pp.3-11
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    • 1992
  • The present study examined various environmental and cultural media conditions for in vitro "rescue" of cross-fertilized ovules formed through sexual crosses between Nicotiana rustica and N. tabacum cv. BY4. The response ovules to two cultural procedures was compared; ovules were cultured either separately or left attached to the placenta. Total yield of seedlings and percent of normal seedlings were increased by culturing individual ovules separately, rather than on excised placenta. Total yield of seedlings and number of normal seedlings were produced following in vitro culture of individual fertilized ovules of N. rustica X M tabacum cv. BY4 at four days post-pollination on NN medium containing 2% sucrose. In the in vitro culture of fertilized ovules, high sucrose concentration increased the frequency of seedlings of abnormal appearance. Therefore, sucrose should be supplied to developing ovules at gradually decreased concentrations. Culture of fertilized ovules from three to eight days after pollination gave increased number of seedlings, but with delayed cultral time the number of morphologically normal seedling were decreased. Hybrids were uniform in appearance and showed vegetative heterosis but flower characteristics were generally intermediate between those of the parents. All hvbrids evaluated were self-sterile.f-sterile.

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