• Title/Summary/Keyword: Cell suspension

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Identification of Salidroside from Rhodiola sachalinensis A. Bor. and its Production through Cell Suspension Culture (참돌꽃에서 Salidroside의 동정 및 현탁세포배양을 통한 분리)

  • Kim, Soo-Jung;Kim, Kwang-Soo;Hwang, Sung-Jin;Chon, Sang-Uk;Kim, Young-Ho;Ahn, Jun-Cheul;Hwang, Baik
    • Korean Journal of Medicinal Crop Science
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    • v.12 no.3
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    • pp.203-208
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    • 2004
  • Salidroside was isolated and purified from R. sachalinensis A. Bor. roots. Purified salidroside was obtained from repeated silicagel column chromatography and preparative HPLC, and identified by $^1H-NMR,\;^{13}C-NMR\;and\;^1H-^1H$ COSY spectra analyzer. Callus induction and cell suspension from R. sachalinensis leaf segments were established on 1/2MS solid medium and in $2B_5$ liquid medium containing 0.5 mg/l NAA and 1 mg/l, BA in the dark condition, respectively. The contents of salidroside for suspension culture were ranging from 0.12% to 0.41% in comparison with 0.17% for natural roots.

Plant Regeneration from Protoplasts of Suspension Cultured Cells in Arabidopsis thaliana (애기장대(Arabidopsis thaliana) 현탁배양세포의 원형질체로부터 식물체 재분화)

  • 김명덕;김준철;진창덕;임창진;한태진
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.2
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    • pp.125-131
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    • 2000
  • Protoplasts of Arabidopsis thaliana were easily isolated from the shoot-forming (SF) suspension-cultured cell clusters with 4 hours-shaking condition (40 rpm) on CPD enzyme solution containing 1% cellulase R-10, 0.25% pectolyase Y-23 and 0.5% driselase. Protoplasts were cultured on liquid KAO medium supplemented with 1 mg/L 2,4-D, 0.5 mg/L kinetin, 200 mg/L spermidine and 68 g/L glucose. Also, protoplasts were cultured on 0.2 $\mu$M membrane filter placed onto CP solid medium containing the suspension cells as feeder cells in the dark at $25^{\circ}C$ for 4 weeks. Protoplast-derived-SF calli were cultured on MS medium containing 0.05 mg/L IAA, 7 mg/L 2 ip and 30 g/L sucrose under the continuous illumination for four weeks. The frequency of shoot formation was about 60%. The regenerants were transferred into potting soil to grow mature plants. The regenerants formed the silques with seeds after 8 weeks of cultures.

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Embryogenic cell suspension culture and plant regeneration in zoysiagrass (Zoysia japonica Steud) (한국들잔디 배아세포의 부유배양과 식물체 재생)

  • Fang, Wenjuan;Han, Liebao;Qi, Chunhui;Li, Deying;Park, Tae-Yun
    • Asian Journal of Turfgrass Science
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    • v.23 no.2
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    • pp.345-352
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    • 2009
  • Zoysiagrass (Zoysia japonica Steud) is a warm season turfgrass species widely used for sports field and golf courses. Many cultivars are propagated through vegetative methods. This study was conducted to develop an optimum culture medium and culture conditions for embryogenic callus induction and plant regeneration, and to establish a cell suspension culture system for use in zoysiagrass breeding and propagation. The results indicated that adding $Cu^{++}$ at 2.5 mg $L^{-1}$ to the induction medium was optimum for callus induction. Increasing the numbers of sub-culture cycles improved the quality of calli. The optimum dosage for cell suspension culture ranged from 2.5 to 10 mL. The embryogenic callus suspension used in this study had a plant regeneration rate of 58%.

Production of ${\gamma}$-Linolenic Acid by Cell Suspension Cultures of Lithospermum erythrorhizon (지치세포 배양에 의한 ${\gamma}$-Linolenic Acid 생산)

  • 김용환;김정봉;류태훈;이철희;황영수
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.2
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    • pp.111-114
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    • 1995
  • To produce ${\gamma}$-linolenic acid (GLA) by cell cultures of Lithospermum erythrorhizon, we optimized medium compositions including carbon sources, nitrogen sources and growth regulators. MS basal medium supplemented with 1.0 mg/L 2, 4-D was effective for callus induction from mesophyll tissue. Addition of sucrose at 88mM concentration induced active proliferation of suspension cells and increased GLA content. Increased supplement of potassium nitrate as nitrogen source resulted in proliferous cell growth and increased total fatty acid content Abscisic acid increased cell growth and fatty acid content in callus culture, whereas as it had an inhibitory effect in suspension cell culture.

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Protoplast Culture and Plant Regeneration of Rice (벼의 원형질체 배양과 식물체 재분화)

  • 이성호;이수인;김주현;코킹 에드워드 씨
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.42 no.3
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    • pp.306-316
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    • 1997
  • Embryogenic calli were induced from mature seed scutella of anther culture-derived rice variety Zhonghua 8. Cell suspension cultures were initiated from friable embryogenic calli and utilized as source material for protoplast isolation. Generally, the older and finer cell suspensions gave higher protoplast yields than younger suspension cultures. Protoplasts exhibited sustained cell division and formed microcalli when cultured in KPR medium supplemented with 0.5 mg $l^{-1}$ 2,4-D, 1.0 mg $l^{-1}$ NAA and 0.5 mg $l^{-1}$ zeatin using the agarose embedding procedure without feeder cells. Protoplast plating efficiencies ranged from 0.20 to 0.54%. Microcalli were transferred to MS medium supplemented with 2.0 mg $l^{-1}$ kinetin and 0.5mg $l^{-1}$ NAA for plant regeneration. The regeneration frequencies were 2 to 12%, depending on the cell suspension lines of Zhonghua 8. The plants were transferred to the glasshouse and were fertile.

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Effect of Suspension Culture Conditions on Cell Activity of Wild Viola(Viola partrinii DC.) Callus (야생 흰 제비꽃(viola patrinii DC.)callus의 현탁배양 방법이 세포 활성에 미치는 영향)

  • Kim, Du-Hyun;Chung, Yong-Mo;Chung, Chung-Han;Yeeh, Yeehn;Kwon, Oh-Chang
    • Journal of Life Science
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    • v.6 no.2
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    • pp.94-103
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    • 1996
  • To understand effect of inoculum size, cell density, sucrose concentration and concentrations of MS basal on suspension culture and protoplast isolation of wild viola(Viola patrinii DC.) callus from petiole segments this experiment was conducted. In the lot of 30 mesh inoculum size, two observations were; One was that a considerable increase in the fresh and dry weight of callus was determined. Another was that the callus mass was relatively compact compared with others. A recommendable cell density was 0.4g for 20ml culture medium and the higher sucrose concentration, the higher fresh and dry weight were obtained. The dilution of MS basal salt was differently affected on fresh and dry weight; the highest fresh weight was found in 1x MS salt, while the higest dry weight was in 1/3x dilution.The addition of casein hydrolysate(3g/L) was more effective to increase of both fresh and dry weight. THe contents of protein was great in the inoculum lots with larger inoculum sizeand higher concentration of MS basal salts contenting 3g/L of casein hydrolysate and higher sucrose compared with others. The greatest protoplasts were isolated from the lot of 10 mesh size treated with 1%pectinase SE-150 and 2% cellulase YC. In general, for optimal protoplast isolation the followingconditions were recommended; 1) Use of smaller cell size cultured for 2-5 weeks, and 2) more than 5 hours incubation using the combined mixture of the enzymes with proper concentrations.

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Establishment of suspension culture condition for embryogenic callus proliferation and somatic embryo development of Kalopanax septemlobus (음나무 배발생 캘러스의 증식 및 체세포배 발달을 위한 액체 현탁 배양조건 확립)

  • Kim, Sun-Ja;Moon, Heung-Kyu
    • Journal of Plant Biotechnology
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    • v.36 no.1
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    • pp.7-12
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    • 2009
  • This study was conducted to establish the optimal suspension culture system for both the propagation of embryogenic cells (ECs) and the induction of somatic embryos (SEs) of Kalopanax septemlobus. The proliferation rate of ECs was reduced as the inoculum density was increased; the highest rate was obtained when 0.1 g/100 ml of cells was initially inoculated. According to the analysis of cell growth pattern and cell growth cycle (G1, Sand G2/M), the cell growth started in 5 days culture initiation, grew rapidly until 15 days and then decreased gradually. Distinctive changes of the cell growth cycle by the culture periods was also observed; the growth cycle was doubled from initial 5.6% to 11.7% of S stage in 5 days culture and then reached in stable stages again. Therefore, the results indicated that a 15-day-cycle was the optimal culture period for the propagation of the ECs through the suspension culture. Furthermore, the cell inoculum density was also important for the induction of SE; more than 65% of SEs at the torpedo stage was induced by using the low level of cell inoculum (0.5 g/L), while the higher inoculum densities were rapidly reduced the proportion of SEs at that stage. Although the higher inoculum density delayed the development of SE, it did not affect the proportion of SEs at the globular and heart stage. In conclusion, this study showed that the suspension culture of the Kalopanax septemlobus ECs through the control of inoculum density was an efficient way for both the propagation of ECs and the induction of SEs, suggesting that the development of this system might help to reduce the culture period for the somatic embryo production.

The Various Suspension Culture Methods on the Growth of Culture Cells of Wild Viola (Viola patrinii Dc.) (야생 흰 제비꽃 (Viola patrinii DC) 배양세포의 활성에 미치는 다양한 현탁배양 방법)

  • 임현희;정용모;조영수;정정한;서정해;권오창
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.3
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    • pp.155-161
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    • 2000
  • This experiment was carried out to examine the effects of culture medium on cell growth of the viola (Viola patrinii DC.) suspension culture. The results are as follows: The greatest cell growth rates were found with MS medium suggesting that this medium could be recommendable for the viola suspension cell culture. When the nitrogen sources (NH$_4$NO$_3$ and KNO$_3$) of MS salts were diluted at half concentrations, the cell growth rates were slightly increased, but when the combined concentration rations of NH$_4$+ and NO$_3$ions were 25 to 75 the greatest cell growth rates were obtained. This result imply that the nitrogen ion sources had slight influence on the rates. Another feature was obtained. This result implys that the nitrogen ion sources had slight influence on the rates. Another feature was that as the concentration of NH$_4$+ ion lowered, the callus color changed to pale yellow with some red spots. The addition of casein hydrolysate (5 g/L) was more effective for the cell growth. On the basis of microscopic observation, the highest cell growth rates were detected during 2-4 weeks culture and after 6 weeks of the culture, some elongated and vacuolated cells were determined.

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Effects of Pluronic F-68 on Cell Growth of Digitalis lanata in Aqueous Two-Phase Systems

  • LEE , SANG-YOON;KIM, DONG-IL
    • Journal of Microbiology and Biotechnology
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    • v.14 no.6
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    • pp.1129-1133
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    • 2004
  • The effects of Pluronic F-68, a non-ionic surfactant, on the growth and physical characteristics of Digitalis lanata suspension cultures were investigated in aqueous two-phase systems (ATPSs) composed of $4.5\%$ polyethylene glycol (PEG) 20,000 and $2.8\%$ crude dextran. In the range of 0.1-10.0 g $1^{-1}$, Pluronic F-68 enhanced the maximum cell density in a medium with ATPSs, even though Pluronic F-68 did not affect cell growth in a normal growth medium. In terms of physical properties of ATPSs with cell suspension cultures, 0.2 g $1^{-1}$ of Pluronic F-68 reduced viscosity by up to $40\%$, while 0.1 g $1^{-1}$ of Pluronic F-68 significantly enhanced the oxygen transfer rate. In addition, we successfully performed aqueous two-phase cultivation in a 5-1 stirred tank bioreactor with 0.5 g $1^{-1}$ of Pluronic F-68, and discovered that cell growth in ATPSs was similar to that in normal growth medium.