• Title/Summary/Keyword: Cultures hairy roots

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The Production of Anti-cancer Substances by in vitro Grown Cultures of Panax ginseng C.A. Meyer

  • Yang, Deok-Chun;Park, Kyung-Hwa;Kim, Yong-Hae;Yoon, Eui-Soo;Kang, Tae-Jin;Park, Kwang-Tae
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 1999.10a
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    • pp.46-57
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    • 1999
  • Ginseng(Panax ginseng C.A. Meyer) is important medicinal plant but requires 4-year cultivation for root harvest because of slow growth. In contrast, ginseng callus and hairy roots grow vigorously and may Produce the same or more biologically active compounds for human health than natural ginseng roots. Therefore, ginseng callus and hairy roots can be used for commercial purposes. Polyacetylene, one of anti-cancer compounds in ginseng, was not detected in the callus cultured on the medium containing 2, 4-B, but cells derived from the callus growth was excellent, The ginseng calli cultured on the medium containing 2mg11 CPA and 0.05mg/1 BA was grown vigorously and produced panaxydol, one of ginseng polyacetylene. The biosynthesis of polyacetylene in callus was not affected by addition of NAA and sucrose in media. The SH medium was better than the MS medium for ginseng callus growth and biosynthesis of panaxydol. Another ginseng anti-cancer compounds, ginsenoside-Rg$_3$, Rh$_1$and Rh$_2$ were detected in ginseng hairy roots by heat treatment. Those of Panax ginseng were obtained after root disks of three-year old roots were infected with Agrobacterium rhizogenes Rl000 $A_4$T in dark condition after one month of culture. The optimum growth of hairy roots was achieved in the culture of 1/2 MS liquid medium in dark(22$^{\circ}C$) under 60 rpm gyratory shaking. Hairy roots grew well in 5 ι Erlenmeyer flasks, 1ι roller drums, 10ι jar-fermenters, and especially in 20ι air-lift .culture vessels. All heat treatments had remarkably different ginsenoside contents. Eleven ginsenosides were determined in heat treatment, eight in freeze dried hairy roots. Contents of ginsenoside-Rbl , Rb2, Rc, Rd. Re, Rf, and Rg$_1$tested in all heat treatments were less than those of freeze dried hairy roots. Contents of glnsenoside-Rg$_2$ in heat treatment for 1 hour at 105$^{\circ}C$ was 4.92mg/g dry wt, 3.9 times higher than 1.27 mg/g dry wt of freeze dried hairy roots. The optimum condition of heat treatment for the production of ginsenoside-Rg$_3$and Rhl was 2 hours at 105$^{\circ}C$, and ginsenoside content was 2.58mg/g dry wt and 3.62mg/g dry wt, respectively. The production of ginsenoside-Rh2 was the highest in heat treatment for 2 hours at 105$^{\circ}C$ among treatments examined, and ginsenoside-Rh$_2$content was 1.08mg/g dry wt.

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Production of Tropane Alkaloids by Hairy Root Cultures of Scopolia parviflora (미치광이풀(Scopolia parviflora)의 모상근 배양에 의한 Tropane Alkaloid 생산)

  • 안준철
    • Journal of Plant Biology
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    • v.36 no.3
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    • pp.225-231
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    • 1993
  • Transformed hairy roots of Scopolia parviflora, producing tropane alkaloids and native to Korea, were obtained following infection of rhizome segments with Agrobacterium rhizogenes A4. Each root tip induced from inoculum sites excised and cultured in MS agar or liquid medium. About seventy of hairy root clones were established. Among them, several fast growing hairy root clones were examined for alkaliod content. Two dimensional TLC analysis showed that the tropane alkaloid pattern of hairy root was more complicated than that in the rhizome of mother plant. On the other hand, some hairy root clones did not produce scopolamine and hyoscyamine. In HPLC analysis, some hairy root clones yield higher levels of scopolamine and hyoscyamine than those of mother plant rhizome which used for infection. Scopolamine and hyoscyamine were identified by comparison of their retention times and of their spectra data with those of authentic compounds.

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Improvement of Tropane Alkaloid Productivity by Optimization of Sucrose Concentration and Addition of Hydroxyapatite in Hairy root Cultures of Scopolia parviflora (미치광이풀 모상근 배양에서 적정 sucrose 농도 및 hydroxyapatite 첨가에 의한 tropane alkaloid 생산성 향상)

  • An, Jun-Chul;Yang, Sun-Ju;Pyo, Byung-Sik;Choi, Ji-Won;Hwang, Baik
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.1
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    • pp.21-25
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    • 1998
  • The effects of sucrose concentration and some absorbents on growth and tropane alkaloid production in hairy root cultures of Scopolia parviflora were investigated. The maximum effect on growth and tropane alkaloid production in hairy root clone SP11 was obtained for 1/2 B5 medium containing 5% sucrose. The production pattern of tropane alkaloid in hairy roots was some different from that of rhizome of mother plant, particulary showing high littorine contents, which was not found in ordinary roots. Among absorbents examined, charcoal 0.01% and XAD-II 1% made a slight growth promotion effect, whereas the other concentration of charcoal, XAD-II and absorbents (amberlite and chitosan) showed inhibition or no significant effect. The addition of hydroxyapatite enhanced the production of tropane alkaloids significantly than control cultures.

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Effect of Polyamines on Indigo Biosynthesis in Hairy Root Cultures of Polygonum tinctorium Lour. (Polyamine이 쪽 모상근배양에서 인디고 생합성에 미치는 영향)

  • Kim, Jin-Nam;Jang, Hong-Gi;Park, Sang-Un;Ryu, Hwa-Won
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.51 no.spc1
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    • pp.247-250
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    • 2006
  • We herein studied the effect of polyamines (putrescine, spermidine and spermine) on growth and indigo biosynthesis in hairy root cultures of Polygonum tinctorium Lour. Our results revealed that polyamine treatment increased hairy root growth and indigo biosynthesisat all tested concentrations, with the highest growth rate (4.4 g/ flask) and indigo yield (216 ug/g) induced by 70 mg/L putrescine. These results show far the first time that the growth rates and indigo biosynthesis of Polygonum tinctorium hairy roots may be improved by addition of polyamines to the liquid culture medium.

Enhancement of Growth and Secondary Metabolite Biosynthesis: Effect of Elicitors Derived from Plants and Insects

  • Jeong Gwi-Taek;Park Don-Hee
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.1
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    • pp.73-77
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    • 2005
  • Plant-derived natural products have been and will continue to be valuable sources. Elicitors have been employed to modify cell metabolism in order to enhance the productivity of useful metabolites in plant cell/tissue cultures. In this study, several elicitors were used to improve the productivity of useful metabolites and to reduce culture time for archiving high concentration in P. ginseng hairy root cultures. The addition of chitosan, chitosan oligosaccharide and alginate oligosaccharide to the culture of P. ginseng hairy roots caused growth to be inhibited with the increase in elicitor concentration. The usage of the chitosan elicitor and D-glucosamine caused a slight decrease in hairy root growth, whereas total ginseng saponin accumulated slightly with the increase in elicitor concentration. When gel beads were added to the culture medium at the initial period, hairy root growth was enhanced. The maximum growth was 1.35 times higher than that of the control at $1\%$ (w/v). Total ginseng saponin content decreased due to the addition of alginate beads. This would result in consistent diffusion of lower levels of calcium ions during the culture period that promotes biomass growth.

Patterns and Contents of Ginsenoside in Normal Root Parts and Hairy Root Lines of Panax ginseng C. A. Meyer (인삼 뿌리 부위별 및 모상근 세포주간 ginsenoside 양상 및 함량)

  • 양덕춘;양계진
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.6
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    • pp.485-489
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    • 2000
  • The patterns and contents of ginsenosides were examined in normal root parts and hairy root lines of Panax ginseng C. A. Meyer. Ginsenoside-Rb$_1$, -Rb$_2$, -Rc, -Rd, -Re, -Rf, -Rg$_1$, -Rg$_2$ were detected in normal roots and hairy roots of ginseng. The patterns and contents of ginsenosides in that were very difference each other. The contents of total ginsenoside of hairy root (KGHR-1) was 17.42 mg/g dry wt, it's highest compared to others. Ginsenoside contents of hairy root (KGHR-1) was higher on ginsenoside-Rd, Rg$_1$, KGHR-5 was higher on ginsenoside-Rb$_1$, Rg$_1$, and KGHR-8 was higher on ginsenoside-Rd, Re than others. The contents of total ginsenosides on 6 years old ginseng cultured in the field were high in the order of main root, lateral root and fine roots, and content of ginsenosides in fine roots was 3.2 times higher than that in main root. The ratio of ginsenoside-Rg$_1$to total ginsenosides were about 3.43%, 8.68% and 14.18% respectively on fine root, lateral root and main root, it's very lower than that in hairy roots. It is suggested that specific ginsenosides can be produce in cultures of ginseng hairy roots.

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Effect of Electron Acceptor, Electron Transport Inhibitors and Antioxidants on Growth and Ginsenosides Production in Hairy Root Cultures of Panax ginseng C.A. Meyer (인삼모상근의 생장과 Ginsenosides 생성에 미치는 Electron Acceptor, Electron Transport Inhibitors 및 Antioxidants의 효과)

  • Kim Yong-Hae;Choi Kyu-Myung;Yang Deok-Chun;Yun Kil-Young;Yang Deok-Cho
    • Journal of Ginseng Research
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    • v.23 no.4
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    • pp.205-210
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    • 1999
  • The growth of hairy roots were increased $69\%$ by 0.1 mM DCPIP under light conditions. In these conditions, the contents of seven ginsenosides were none significant variation. The influence of electron inhibitors on growth and ginsenoside contents in ginseng hairy roots was tested. The growth was inhibited $71\%\;and\;22\%$ respectively by CCCP and methylamine. The ginsenoside contents were as decreased above $45\%$ in all treatment tested except triazine treatment. In antioxidants treatment, the growth of hairy roots was increased about $68\%$ by propylgallic acid, about $23\~25\%$ by ascorbic acid or 2,5-dimethylfuran, while the contents of seven ginsenosides were none significant variation. The ginsenoside productivity was high when hairy roots were cultured in $\circledR^{\wedge}MS$ medium for 4 weeks and then transferred to 1/2MS medium with ascorbic acid or 2,5-dimethylfuran for 1 weeks in light conditions. It is suggested that ginsenoside productivity could be accelerated by some antioxidants in hairy root cultures of ginseng.

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A New Chemical Constituent from the Hairy Root Cultures of Catharanthus roseus

  • Chung, Ill-Min;Park, Han-Young;Ali, Mohd;San, Ka Yiu;Peebles, Christie A. M.;Hong, Seung-Beom;Ahmad, Ateeque
    • Bulletin of the Korean Chemical Society
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    • v.28 no.2
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    • pp.229-234
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    • 2007
  • One new compound, 3,7,11,19,23,27-hexamethyl-15-hydroxymethylene-n-octacos-5,8,20-triene-10β,18α- diol-10β-D-glucopyranoside (1), along with the three known compounds, 3-epibetulinic acid (2), n-pentadecanyl octa-dec-19-en-oate (3) and β-sitosterol (4) were isolated from the methanolic extract of the cultured Catharanthus roseus hairy roots. The structures of the one new and three known compounds were elucidated using one- and two-dimensional NMR in combination with IR, EI/MS, FAB/MS. To the best of our knowledge, 3,7,11,19,23,27-hexamethyl-15-hydroxymethylene-n-octacos-5,8,20-triene-10β,18α-diol-10β-D-glucopyranoside, 3-epibetulinic acid and n-pentadecanyl octa-dec-19-en-oate were identified for the first time from the hairy roots of C. roseus.

Regulation of Growth and Catharanthine Production by the Intracellular Phosphate Level in Hairy Root Cultures of Catharanthus roseus (세포내 인산농도에 의한 일일초 모상근생장 및 Catharanthine 생산의 조절)

  • Kyung-Hee JUNG;Sang-Soo KWAK;Cha Yong Choi;Jang R. LIU
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.4
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    • pp.183-187
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    • 1995
  • The relationship between the intracellular phosphate level and catharanthine production in hairy root cultures of Catharanthus roseus was investigated The growth of hairy root increased in proportion to the phosphate concentration in the growth medium. When hairy roots were cultured in a phosphate-free medium, the catharanthine content was increased to the highest level As the phosphate concentration in the medium was enhanced, the catharanthine content decreased proportionally. Hairy root cultures with elevated intracellular phosphate concentrations indicated that growth was proportional to the level of intracellular phosphate. Catharanthine production increased abruptly below an intracellular phosphate level of $100\;\mumoles/g$ dry wt. The intracellular phosphate level may play a key role in the regulation of growth and secondary metabolite production.

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