• Title/Summary/Keyword: Plant cell cultures

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Isolation and Characteristics of Brachybacterium sp. SY -97 Killing the Harmful Dinoflagellate Cochlodinium polykrikoides (유해적조생물 Cochlodinum polykrikoides를 살멸하는 Brachybacterium sp. SY-97의 분리 및 특성)

  • Kim, Yun-Sook;Jeong, Seong-Yun;Lee, Sang-Joon;Lee, Won-Jae
    • Journal of Environmental Science International
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    • v.18 no.4
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    • pp.435-443
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    • 2009
  • A bacterial strain SY-97 that showed algicidal activity against Cochlodinium polykrikoides was isolated from coastal water of Uljin (eastern coast of Korea) in August, 2005. The isolated strain was identified as Brachybacterium sp. by morphological and biological tests, and analysis of 16S rDNA sequence. The optimal culture conditions for the growth of strain SY-97 were $30^{\circ}C$, initial pH 7.0, and salinity 2.0%. From the result of cell culture insert experiment, Brachybacterium sp. SY-97 is assumed to produce secondary metabolites which have algicidal activity. When 10% culture filtrate of this strain was applied to C. polykrikoides ($1.2{\times}10^4\;cells/m{\ell}$) cultures, 100% of C. polykrikoides cells was destroyed within 15 hours. The released algicides were heat-tolerant to $100^{\circ}C$ and stable in pH $6.0{\sim}10.0$. These results suggest that Brachybacterium sp. SY-97 is potentially useful for controlling outbreaks of C. polykrikoides.

Optimization of liquid-liquid extraction conditions for paclitaxel separation from plant cell cultures (식물세포배양으로부터 Paclitaxel 분리를 위한 액-액 추출 조건의 최적화)

  • Kim, Jin-Hyun
    • KSBB Journal
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    • v.24 no.2
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    • pp.212-215
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    • 2009
  • In this study, the process parameters of liquid-liquid extraction were optimized to obtain a high purity and yield of paclitaxel in a pre-purification step. The optimal solvent ratio (methylene chloride/concentrated methanol extract ratio), extraction times, mixing time, and standing time for liquid-liquid extraction were 0.28 (v/v), 3(times), 30 min, and 40 min, respectively. The polar impurities from the biomass extraction were efficiently removed by liquid-liquid extraction. The complete concentration of liquid-liquid extract by rotary evaporator was reliable enough to obtain a high purity and yield of paclitaxel for subsequent purification steps.

Decreasing Particle Size of Paclitaxel Using Polymer in Fractional Precipitation Process (고분자물질을 이용한 분별침전 공정에서 파클리탁셀의 입자크기 감소)

  • Kim, Min Jae;Kim, Jin-Hyun
    • Korean Chemical Engineering Research
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    • v.54 no.2
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    • pp.278-283
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    • 2016
  • In this study, we have for the first time applied fractional precipitation with hydrophilic polymer in order to decrease the particle size of the anticancer agent paclitaxel from plant cell cultures. When compared with the case where no hydrophilic polymer was employed, the addition of hydrophilic polymer in fractional precipitation resulted in a decrease in the size of the paclitaxel precipitate. Among the polymers used, HPMC 2910 was the most effective for inhibition of precipitate growth. A polymer concentration of 0.2% (w/v) obtained the smallest particle size. The particle size was reduced by ~35% compared to control. In addition, the precipitate size was inversely correlated with the absolute value of the zeta potential.

Plant Production from Desiccated Somatic Embryos of Acanthopanax chiisanensis (지리오가피 (Acanthopanax chiisanensis) 체세포배의 건조처리를 통한 식물체 증식)

  • Lee, Kang-Seop;Bang, Keuk-Soo;Choi, Yong-Eui;Ahn, Byung-Yong
    • Journal of Plant Biotechnology
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    • v.30 no.4
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    • pp.381-385
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    • 2003
  • An efficient method of plant regeneration from Acanthopanax chiisanensis somatic embryos was developed. Cotyledonary somatic embryos were obtained in liquid Murashige and Skoog (MS) medium from embryogenic cell suspension cultures. They were desiccated for 0 to 72 hr and then cultured on MS medium containing NAA, BA, GA$_3$, (0-0.5mg/L). The highest multiple shoots formation (100%) was obtained from 72 hr desiccated somatic embryos on ifs medium with 0.5mg/L NAA+0.5mg/L BA or 0.5 mg/L NAA+0.5mg/L BA+0.5mg/L GA$_3$ after 6 weeks culture. Plant conversion from multiple shoots was not high. The highest plant conversion from multiple shoots was obtained on 1/3MS medium with 1.0mg/L GA$_3$. Converted plantlets were transferred to ex vitro condition and the highest survival rate (70%) of the plantlets was obtained on plastic pots containing vermiculite and sand. These results indicate that micropropagation procedure can be applied for an efficient mass propagation of Acanthopanax chiisanensis.

Cultures of Ginkgo biloba, Effect of Nutritional and Hormonal Factors on the Growth of Cultured Cells Derived from Ginkgo biloba

  • Jeon, Mee-Hee;Sung, Sang-Hyun;Jeon, Soon-wha;Huh, Hoon;Kim, Jin-woong;Kim, Young-Choong
    • Archives of Pharmacal Research
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    • v.16 no.3
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    • pp.244-250
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    • 1993
  • Calli and suspension cultures were obtained following inoculation of the explant from leaves of Ginkgo biloba L on the supplemented MS basal medium. The obtained calli and suspension cultured cells were able to produce detectable amounts of ginkgolides which are known as natural specific PAF antagonists. The production of ginkgolides in the calli and suspension cultured celles were identified using GC/MS, GC and HPLC with authentic ocmpounds. Since the production of ginkgolides A and B the calli and suspension cultured cells had been confirmed, effects of types and concentration of plant growth regulators, media and illumination on the induction and growth of the callus were studied. The concentrations of growth regulators for optimal callus were studied. The concentrations of growth regulators for optimal callus induction were studied. The concentrations of growth regulators for optimal callus induction were 1.0 to 2.0 mg/L for NAA and o.1 mg/L for kinetin. The growth of the Callus seemed to be more simnultaed with the combination of NAA and kinetin than NAA and BA with illumination at all concentration ranges of 1.0 to 4.0 mg/l for NAA and o.1 to 1.0 mg/L for kinetin or BA studied. Amogn 8 different media used, the induction rate of callus on Anderson, Eriksson, and Shenk and Hildebrant at 4 weeks after the innoculation was almost the same as that of MS. However, callus was rarely induced on Heller or White medium. Suspension cultures were easily initiated with 3 g of callus (fresh weight) derived from ginkgo leaves on supplemented MS medium. A typical growth curve of suspension cultured cells could be obtained by measuring the fresh weight of the suspension cultured cells at every 3 days. To improve the growth of suspension cultured cells of ginkgo, effects of concentrations of NAA, sucrose, phosphate ions and molar ratio of $NH_{4}^+\;to\;NO_{3}^-$ ions in the culture medium were studied. The maximum growth of the cells was achieved when the culture medium contained 1.0 mg/L of NAA, 30 g/L sucrose, 1.75 mM phosphate ions and 1:5 molar ratio of $NH_{4}\;to\;NO_{3}^-$ ions.

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Scopolamine Production in Suspension Cultures of Tumor Calli from Datura metel L. (흰독말풀(Datura metel L.)종양 캘러스의 현탁배양으로부터 Scopolamine 생성)

  • 이수경;윤길영;김용해;양덕조
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.3
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    • pp.203-211
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    • 2000
  • In this study to produce large-scale scopolamine we were examined in the tumor calli of Datura metel L. induced by Agrobacterium tumefaciens $Ery{101}$. The growth and scopolamine contents of tumor calli were higher under light condition than in dark. The optimum condition of growth and scopolamine production were fluence rate of 16 $\mu$mol $m^{-2}s^{-1}$, spectra of red light region and 16 hour light periods on 50 mL SH liquid medium in 4 weeks culture. To increase of the scopolamine contents in tumor calli, the optimum concentration of nitrogen source were 1.8 mM NH$_4$+ and 40 mM NO$_3$. The optimum elicitor concentration for production of scopolamine were 10 mg/L chitosan and 15 mg/L yeast extract. The effect of precursors were good at the concentration of 0.2 mM tropine and 0.3 mM tropic acid, respectively. In order to increase of growth and scopolamine contents. we induced mutant from Datura metel L. tumor callus. Mutants of tumor calli were obtained by 3 Krad, 4 Krad and 6 Krad of ${60}^Cor-ray$. Among them, 3 Krad tumor callus was excellent on the growth and teratoma induction. The 4 Krad tumor callus was negligible for both growth and teratoma induction. But the 6 Krad tumor callus was the best in growth and teratoma induction. The formation of the mutant calli can be enhanced through hormonal combination of 1 mg/L 2,4-dichlorophenoxyacetic acid and 0.5 mg/L benzyladenine. We carry out selection mutant tumor calli for high content tropane alkaloid and suspension cultures for scopolamine production.

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Characterization of Anionic Peroxidase Induced by Low Host-Specific Elicitor in Suspension Cultures of Rose (Rosa sp.) (장미(Rosa sp.) 현탁배양세포에서 숙주 특이성이 낮은 Elicitor에 의해 유도되는 Anionic Peroxidase의 특성)

  • 신미선;양은진;이인철
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.4
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    • pp.277-282
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    • 1998
  • Whereas cationic extracellular peroxidases (PODs) were observed in the suspension cultures of rose (Rosa sp. L. cv Pual's scarlet) grown under normal conditions, new anionic isozymes were induced within 24 hr by the treatment of low host-specific elicitor (10 mg glucan/L media) prepared from yeast cell wall. Prominent anionic (pI 6.1) and cationic POD (pI 8.4) were purified and characterized to understand the physiological role of the enzymes. Both enzymes were purified (ca.200 fold) by the ammonium sulfate precipitation, ion exchange chromate-graphy and gel filtration chromatography. The Km values of the purified anionic POD for ferulic acid and $\textrm{H}_2\textrm{O}_2$ were 4.64 mM and 0.72 mM, whereas those of the cationic POD were 1.38 mM and 0.48 mM, respetively. The activity of the anionic POD as NADH oxidase was twice higher than that of cationic POD. The NADH oxidation in the anionic POD fraction was inhibited by 60% on the addition of 0.1 mM coniferyl alcohol, while that in the cationic fraction was inhibited by 15%.

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Plant Regeneration and Protein Analysis from Cadmium Resistant Callus of Tobacco (Nicotiana tabacum cv. BY4) (담배 (Nicotiana tabacum cv. BY4)카드뮴 저항성 캘러스로부터 식물체 재생과 단백질 분석)

  • 오승철;소웅영;조덕이;양덕춘
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.1
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    • pp.7-13
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    • 2001
  • Calli were induced from diploid and haploid tobacco after 4 weeks and maintained on MS medium with combination of 2.0 mg/L 2,4-D,0.1 mg/L BAP and 2.0 mg/L kinetin. Suspension cells were screened through 65 $\mu$m-nylon mesh and 100 $\mu$m-mesh, then they were smeared on selection medium combined with cadmium and PFP by using the low melting agarose of 0.8%. After 30days smeared cultures of the medium the cell was treated with 500 $\mu$M and 1000 $\mu$M to select the resistant cell line were selected. Plant regeneration was induced from the selected cell lines on medium with 0.5, 1.5, 2.0 mg/L BAP and on media with combination of auxin and BAP under 500 $\mu$M and 1000 $\mu$M cadmium. At this time, plant regeneration was achived on cadmium free medium. In case of haploid, occurred from the cell line which is selected in medium with cadmium and PFP. In case of diploid regeneration occurred is in the medium with cadmium alone. The plantlet regenerated from cadmium resistant calli grew well in cadmium 500 $\mu$M. Protein pattern of leaf, root, stem of regenerated plants was analyzed. The quantum was 6.5188 ug/mg.fr.wt in the leaf of plant, 5.3611 ug/mg.fr.wt in the stem, 3.0213 ug/mg.fr.wt in the root. On the other hand, 5.9652 ug/mg.fr.wt. in the leaf of control, 3.5974 ug/mg.fr.wt in the stem of the control, 4.3766 ug/mg.fr.wt. in the root of the control. The one dimension bends regenerated from cadmium resistant calli resistant to cadmium in leaf were 49 involving 198.7KD etc. Disappeared were 4 involving 160.5KD etc, The protein bends were combinized were 3 involving 83.4KD etc. The bends resistant to cadmium stress in stem were 41 involving 4.3KD etc. Disappeared were 5 involving 114.8KD etc. The protein bends combinized were 6 involving 128.7KD etc. The bends which had the resistance to cadmium stress in root is 27 in volving 166,9KD etc. The bends which disappeared were 198.7KD etc. There were 5 involving 83.4KD etc.

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Expressions of A-type and C-type Cyclins Induced by Exogenous Cytokinin Treatment on Leaf Blades and Calli of Rice (Oryza sativa L.) (벼의 엽신 및 캘러스에서 Cytokinin 유도성 A-type 및 C-type Cyclin 유전자의 발현 분석)

  • Lee Honggun;Choi Seungho;Hwang Hyunsik;park Jungan;Lee Taekkyun;Park Jongbum;Auh Chungkyoon;Lee Sukchan
    • Journal of Plant Biotechnology
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    • v.32 no.1
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    • pp.15-21
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    • 2005
  • The expression patterns of cyclin genes, which play a crucial role on cell cycle control, were analyzed with rice calli and leaf blades from seedlings. When callus was transferred from media containing the combinations of 2,4-D and kinetin under the dark conditions to medium supplemented with cytokinin-only on 7 days after the cultures, the expression levels of A-, B- and C-type cyclins from callus were increased significantly. Despite the fact that cyclin genes were well expressed on leaf blades rather than other organs in rice seedlings, rice leaf blades grown on the medium containing various combinations and concentrations of cytokinin for 24 hours had no major effect on the expression patterns of cyclins except zeatin. The relation between cytokinin regulation and the expression of cyclins of rice is discussed.

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

  • Park, Soo-Young;Ryu, Sun-Hwa;Kwon, Suk-Yoon;Kim, Jong-Guk;Kwak, Sang-Soo
    • Journal of Plant Biotechnology
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    • v.30 no.2
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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.