• 제목/요약/키워드: two stage cultures

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냉동보존이 진주조개(Pinctada fucata martensii) 유생의 미세구조에 미치는 영향 (The Effects of Cryopreservation on Fine Structures of Pearl Oyster(Pinctada fucata martensii) Larvae)

  • 최윤희;조필규;김태익;배승철;장영진
    • 한국발생생물학회지:발생과생식
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    • 제11권2호
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    • pp.79-84
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    • 2007
  • 냉동보존이 진주조개(Pincata fucata martensii) 유생의 형태 및 구조에 미치는 영향을 알아보기 위하여 냉동전후 D형 및 각정기 유생을 광학 및 전자현미경으로 조사하였다. 동해방지제는 0.2 M sucrose를 첨가한 2.0 M $Me_2SO$를 사용하였다. 냉동후 유생은 일부 패각이 손상되긴 했지만 hinge와 prodissoconch가 뚜렷하게 나타났으며, 소포체, 지질 과립, 미토콘드리아, 핵 등을 포함한 세포내 소기관들이 고르게 분포되어 있었다. 또한, 섬모가 규칙적으로 배열되어 있었고, 섬모 아래 미토콘드리아와 지질과립이 위치해 있는 것이 관찰되었으나, 일부 해동된 유생에서 섬모의 불규칙적인 배열과 섬모환이 둥글게 뭉쳐져 있는 모습이 관찰되었다. 이러한 결과는 진주조개의 D형 유생과 각정기 유생이 냉동에 쉽게 영향을 받을 수 있다는 것을 보여준다. 따라서 냉동보존 시 세포의 손상을 감소시킬 수 있는 연구가 이루어져야 할 것으로 사료된다.

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더덕의 체세포배발생에서 전형층분화와 자엽수의 관계 (Relationship of Cotyledon Number with Procambium Differentiation in Somatic Embryogenesis of Codonopsis lanceolata L.)

  • 최필선;소웅영;조덕이;유장렬
    • Journal of Plant Biotechnology
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    • 제32권2호
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    • pp.135-138
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    • 2005
  • 더덕 자엽절편을 1 mg/L 2,4-D가 첨가된 MS기본배지에 배양하여 배발생캘러스를 얻었고, 호르몬이 첨가되지 않은 MS액체배지에서 구형기의 체세포배를 얻었다. 구형기에서 초기 심장형기로 발달할 때 자엽의 시원세포는 전형성층조직으로부터 분화되기 시작 하였으며, 하배축에서 관찰되는 원통형 전형성층대는 2개의 자엽을 형성할 경우 2개의 전형성층대가, 3개의 자엽은 3개의 전형성층대가, 4개의 자엽은 4개의 전형층대가 각각 독립적으로 분화되어 자엽절과 자엽부위까지 연결되어 있었다. 만약 구형기 체세포배에서 자엽시원세포가 원통형으로 분화될 경우 합생자엽을 갖는 체세포배가 형성되었다.

Distinctive response of maize (Zea mays L.) genotypes in vitro with the acceleration of phytohormones

  • Muppala, Sridevi;Gudlavalleti, Pavan Kumar;Pagidoju, Sreenu;Malireddy, Kodandarami Reddy;Puligandla, Sateesh Kumar;Dasari, Premalatha
    • Journal of Plant Biotechnology
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    • 제47권1호
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    • pp.26-39
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    • 2020
  • In maize, immature embryos (IEs) are highly regenerative explants most suitable for producing high frequencies of plantlet regeneration in vitro. Apart from media, explants, and hormones, genotypic variation also influences in vitro characters to a great extent. In the present study, IEs were used to study the distinctive effect of variation of size/stage and hormones in different genotypes on five in vitro characters viz., frequency of callus induction, growth rate of total callus, frequency of E. callus induction, and volume and number of regenerated plantlets. LS medium with different concentrations of 2,4-D (0.5, 1.5, 2.5, 4.0 and 5.0 mg/L) were used to study the former four in vitro characters, and medium with 6-benzylaminopurine and kinetin (0.5 mg/L, each) was used for plantlet regeneration. IEs of 1.0, 1.5, 2.0, 2.5 and 3.0 mm in size were isolated from four inbred lines viz., NM74C, NM81A, NM5883 and NM5884. Two-way ANOVA revealed that explant size and genotypes, as well as hormonal concentrations showed significant effects on in vitro characters. Two millimeter IEs were found to be suitable for in vitro cultures. LS medium with 1.5 mg/L 2,4-D and LS with BAP and Kn (0.5 mg/L, each) were found to be the best hormonal concentrations for callus induction, maintenance, and regeneration, respectively. Among the four genotypes, NM81A and NM5883 yielded more non-embryogenic and Type I E. calli. In contrast, NM74C and NM5884 yielded more highly regenerative Type II calli. Inbred line NM5884 was found to be the best among these four genotypes.

Poxvirus 감염(感染)에 있어서의 Virus-숙주세포(宿主細胞)의 상호관계(相互關係) 1. Cowpox Virus-FL 세포계(細胞系)의 세포화학적(細胞化學的) Autoradiography 및 세포면역학적해석(細胞免疫學的解析) (Studies on Host-Virus Interaction of Poxviruses 1. Cytochemical, Autoradiographic and Immunocytological Analysis in Cowpox Virus-FL Cell System)

  • 김우호
    • 대한수의학회지
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    • 제15권1호
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    • pp.57-67
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    • 1975
  • The poxvirus group is considered to be a typical cytoplasmic inclusion forming virus. Every poxvirus has been reported to produce only one kind of inclusion in the infected tissues. A vague concept that inclusions of poxviruses are eosinophilic or acidophilic has prevailed. Although many papers and theories about the nature of the inclusion have been presented, most of them are not quite convincing on the point of the relations with virus multiplication, and an analysis of papers published showed that there seem to be many discrepancies in the descriptions of the nature of the poxvirus inclusions. Comparative studies on host-virus interaction with cowpox, orf, swinepox and fowlpox viruses which selected from each Group (I-IV) of poxviruses were performed from the morphological and virological standpoints. At first, in cowpox virus-FL cell system, as a comparative model, cytoplasmic inclusion, nucleic acid metabolism by autoradiography and detection of viral antigen by immunofluorescence were studied and obtained the results as follows: 1. The focus-like cytopathic effect (CPE) at early stage developed to entire culture at terminal stage of infection, and also the developing status of CPE was correlated to viral doses for inoculation. Two kinds of cytoplasmic inclusions which named A and B type were easily observed by Giemsa, hematoxylin-eosin (H & E) and May-Greenwald Giemsa (MGG) stainings in the infected cells. The B type inclusions were formed at early stage of infection and the A type inclusions were produced subsequently the B type formation. The B type which common type inclusion in poxviruses was a small compact or aggregate at early stage and developed to a large diffuse body at terminal stage of infection. On the other hand, the A type inclusion which depend upon the kind of virus was appeared as round and discrete shape, and its size and number was increased gradually during the culture period. It was characteristic to form distinct halos around the both types of inclusions in acid fixed, H & E stained preparations of infected cultures. The B type inclusion was always positive in Feulgen reaction and showed as DNA containing body but the A type inclusion was not. 2. In the relationship between inclusion and DNA metabolism of infected cells by the qualitative autoradiography using 3H-thymidine, the appearance of silver grains was coincided with B type inclusion but not with A type inclusion. This showed that the DNA synthesis was proceeded in all B type inclusions except those in the terminal stage with a diffuse form. This suggested that the B type inclusions are only sites of DNA synthesis and this was proceeded after the cell infection independently. The activity of DNA synthesis of the inclusions was nearly the same as that of the nucleic of normal cells and non-inclusion bearing cells. and non-inclusion bearing cells. Regardless of the size of the degree of DNA synthesis of the B type inclusion, inclusion bearing cells all showed remarkable suppression of nuclear DNA synthesis. 3. By the direct fluorescent antibody technique viral antigen in infected cells was detected. The B type inclusions have been proved to contain a great deal of viral antigen, whereas the basic substance of A type inclusion did not show antigenicity except the round edge. It was suggested that the round edge fluorescence might be caused by the glare of cytoplasmic viral antigen which pushed out and concentrated by the A type inclusion development. 4. Hemorrhagic red pock formations on chorioallantoic membrane of embryonated chicken egg had proved the characteristic of used viral strain. 5. By the above studies on the nature of two types of inclusions and the role they play in virus multiplication, it was concluded that the B type inclusion must be the site of the synthesis of viral DNA and protein as well as the site of the virus.

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콩과 벼 현탁배양(懸濁培養) 중 PCP 수용성대사물(水溶性代謝物)의 동정(同定);3. PCP glucose conjugates의 동정(同定) (Identification of Water Soluble Metabolites of Pentachlorophenol(PCP) in the Suspension Cultures of Soybean and Rice Cells;3. Identification of PCP Glucose conjugates)

  • 김필제;박창규
    • 한국환경농학회지
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    • 제15권2호
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    • pp.167-178
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    • 1996
  • 전보에서 대두(大豆)와 벼의 현탁배양세포(懸濁培養細胞)에서 생성된 PCP의 주요 수용성대사물은 PCP를 비롯한 여러 종류의 염화페놀과 glucose간 ${\beta}-linkage$로 연결된 ${\beta}-glucoside$ 구조를 가질 것으로 추정한 바 있다. 본 연구에서는 conjugates를 가수분해시키지 않는 조건에서 기기적인 방법으로 그 구조를 확인하고자 하였다. Acetyl화(化) 한 glucose conjugates는 aglycones 분석에서 예상한 대로 3종류 이상이 존재하는 것으로 확인할 수 있었으며, 충분히 정제한 각각의 acetyl화(化) 대사물을 FAB-MS로 분석한 결과 분자량과 aglycones part와 관련된 특징적 spectral data 등 conjugates 구조 확인에 필수적인 정보를 얻을 수 있었다. 이러한 구조적 정보와 지금까지의 연구 결과, 즉 현탁배양초기에 생성된 PCP의 glucose conjugates는 ${\beta}-glucosidase$에 의해 특징적으로 가수분해되고, aglycones이 PCP 외에도 tetrachlorophenol 및 tetrachlorocatechol 등이라는 사실을 종합할 때 현탁배양에서 생성된 conjugates는 주로 pentachlorophenyl ${\beta}-D-glucopyranoside$, tetrachlorophenyl ${\beta}-D-gluco$ pyranoside 및 2-hydroxy-3,4,5,6-tetrachlorophenyl ${\beta}-D-glucopyranoside$인 것으로 확인할 수 있었다.

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Large-scale Culture of Plant Cell and Tissue by Bioreactor System

  • Son, Sung-Ho;Park, Sung-Mee;Park, Seung -Yun;Kwon, Oh-Woung;Lee, Yun-Hee;Paek, Kee-Yoeup
    • Journal of Plant Biotechnology
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    • 제1권1호
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    • pp.1-7
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    • 1999
  • Large-scale cultures of plant cell, tissue, and organ have been achieved by using BTBB. When different sized BTBBs (5 L, 20 L, 100 L, 300 L, and 500 L) were tested for the culture of yew cells (Taxus cuspidata Sieb. et Zucc.), cell growth increment reached to 94.5% in SCV after 24 days of culture with 30% of inoculation cell density. However, there were some variations in the production of taxol and its derivatives among the BTBBs of different size. Approximate 4 ㎎/l of taxol and 84 ㎎/l of total taxanes were obtained by using a 500L BTBB after 6 weeks of culture. With a 20L BTBB, about 20,000 cuttings of virus-free potatoes (cv. Dejima) could be obtained by inoculating 128 explants and maintaining 8 weeks under 16 hr light illumination. The frequency of ex vitro rooting of the cuttings revealed as more than 99% under 30% shade. By incorporating two-stage culture process consisting of multiple bulblet formation in solid medium and bulblet development in liquid medium, mass propagation of lily through bioreactor seemed to be possible. In the case of 'Marcopolo', the growth of mini-bulblets in BTBB was nearly 10 folds faster than that of the solid medium. Time course study revealed that maximum MAR yield of ginseng (Panax ginseng C. A. Meyer) in a 5 L and 20 L BTBB after 8 weeks of culture was 500 g and 2.2 ㎏, respectively. By cutting the MAR once and/or twice during the culture, the yield of root biomass could be increased more than 50% in fresh weight at the time of harvest. With initial inoculum of 500 g of sliced MAR in a 500 L BTBB, 74.8 ㎏ of adventitious root mass was obtained after 8 weeks of culture. The average content of total ginseng saponin obtained from small-scale and/or pilotscale BTBBs was approximately 1% per gram dry weight. Based on our results, we suggest that large-scale cultures of plant cell, tissue, and organ using BTBB system should be quite a feasible approach when compared with conventional method of tissue culture.

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활성슬러지내 Nocardia 거품현상 진단을 위한 Fatty Acid Methyl Ester (FAME) 기술 (Fatty Acid Methyl Ester (FAME) Technology for Diagnosing Nocardia Foaming in Activated Sludge)

  • 이재우;김일규;이석헌;안규홍
    • 상하수도학회지
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    • 제18권4호
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    • pp.480-485
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    • 2004
  • Fatty acid methyl ester (FAME) technology was evaluated as a monitoring tool for quantification of Nocardia amarae causing a nuisance foaming problem in activated sludge process. The identified signature peak was 19:1 alcohol as a reliable unique peak to N. amarae. Chemostat study revealed that the distribution and quantity of fatty acid peaks were dependent on the growth stage of Nocardia. The FAME results were similar for two relatively high dilution rates; however, the amounts of signature peaks extracted from the 4 and 6 day cultures were significantly higher. This dependence of signature peaks on the physiological state of the organism may be a useful information to assess the health of microbial populations in activated sludge. A laboratory scale batch foaming potential experiment provided a critical foaming level depending on Nocardia population. This critical Nocardia level determined in this study was in terms of either the threshold filament intersections number or the threshold signature FAME amount. The threshold peak area of signature FAME (19:1 alcohol) and corresponding filament counts were 430PA/mg VSS and $1.45{\times}10^6$ intersections/g VSS, respectively. The threshold signature FAME level could be effectively applied as a criterion for diagnosing foam occurrence in activated sludge system.

쌀보리 약배양을 위한 약치상 방법별 배양효율 (Anther Culture Efficiency According to Plating Method in Naked Barley)

  • 박태일;정선옥;김영진;김현순;서재환;박기훈;김정곤;윤성중
    • Journal of Plant Biotechnology
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    • 제34권3호
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    • pp.237-242
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    • 2007
  • Barley anther culture is hard working to plating picking out anther from the glume and demand long time comparing to be short available development stage for effective culture. Also, it has been treatment massive materials due to low plantlet comparing to get desirable plants intensively. Consequently, this experiment was carried out trying to be more high barley anther culture effectively in terms of save plating effort. Plating materials and culture temperature affected anther culture efficiency are among the inoculation tissues or organs such as anthers, spikelets and whole panicles, culture efficiency was higher with spikelets in two-rowed than six-rowed barley due primarily to a lower contamination, and calli were induced within 30 to 50 days. Callus induction and plant regeneration rates were higher in cultures at $25^{\circ}C$ than at $15^{\circ}C$ and $20^{\circ}C$. Days to callus induction were 25 to 50 days at $25^{\circ}C$ and 50 to 60 days at $20^{\circ}C$.

한국 코스프레 집단의 문화기술지적 연구 I -한국의 코스프레 집단의 분석 - (An Ethnographic Study on CosPlay Group in Korea I - Analysis on CosPlay Groups in Korea -)

  • 고애란;신미란
    • 복식문화연구
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    • 제13권6호통권59호
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    • pp.919-933
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    • 2005
  • This study goes into the field of CosPlay that takes place in Korea, and directly observes the people who engage in CosPlays. Based on in-depth interviews, this research identified the reasons why the CosPlay aficionados participate in this practice and their cultural preferences. Ethnography methodology was used to understand the behavior of the cultural entities of CosPlays. Moreover, this research attempted to understand their daily formalities through their own perspective and language instead of superficial language. In general, CosPlay form that they manifest is classified into two main categories: those who enjoy posing by becoming their favorite characters and those like to wear the clothes of their favorite characters and go up on the stage to perform an act of animation or game where the applicable character appears. Some of these groups are: 1) 'Bishojo CosPlay' that CosPlays mostly Bishojo cartoon characters and wants to just stand out, 2) 'Aekyo' group that CosPlays merely for the sake of the enjoyment of wearing costumes instead of having the mania-like characteristics, 3) 'CosPlayer Group' who enjoy doing organized team CosPlay in order to present a performance, 4) 'J-Rock CosPlay' group that CosPlay Japanese visual rock groups and prepare to be among a professional CosPlay team, and 5) 'People related to Cospre.com' who try to make CosPlay rooted in as one of new cultures.

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고구마 배양세포에서 Peroxiredoxin cDNA의 분리 및 발현 특성 (Molecular Cloning and Characterization of a Peroxiredoxin cDNA from Cell Cultures of Sweetpotato)

  • 박수영;류선화;권석윤;김종국;곽상수
    • Journal of Plant Biotechnology
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    • 제30권2호
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    • pp.135-141
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    • 2003
  • Peroxiredoxin(Pix) are large family of peroxidases that reduce alkyl hydroperoxides and hydrogen peroxide. A cDNA clone (referred to as swPrxl) encoding Pix was from a sweetpotato cDNA library constructed from suspension-sultured cells, and its expression was investigated in terms of stress. The swPrxl contained an open reading frame (ORF) encoding mature protein of 193 amino acids with calculated molecular mass of 20.8kDa. The predicted amino acid sequence of swPrxl has two conserved cysteines that are essential resicues for the reduction of peroxides. It showed high amino acid sequence homology ot PixIIF of Arabidopsis (77%) and putative Prx of rice(72%). RNA gel-blot analysis showed that swPrxl gene was expressed dominantly in leave among intact tissues, and also highly detect in suspension-cultured cells. Interestingly, the level of swPrxl transcripts was almost the same regardless of the growth stage in suspension culture. Furthermore, the transcription level of swPrxl gene was not significantly changed in response to various stress treatments such as wounding, extreme temperature and stress-related chemicals RT-PCR analyses.