• Title/Summary/Keyword: Hairy root culture

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Induction of Sesquiterpene Cyclase During Integrated Extraction of Sesquiterpenes from Hairy Root Cultures of Hyoscyamus muticus (Hyoscyamus muticus의 모상근배양으로부터 Sesquiterpene 화합물의 Intergration 추출시 Sesquiterpene Cyclase의 유도)

  • BACK, Kyoungwhan;SHIN, Dong Hyun;KIM, Kil Ung;De HAAS, Cynthia R.;CHAPPELL, Joseph;CURTIS Wayne R.
    • Korean Journal of Plant Tissue Culture
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    • v.24 no.5
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    • pp.273-277
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    • 1997
  • The sesquiterpene cyclase (SC) was induced and its products were accumulated in the culture media of Hyoscyamus muticus hairy roots by addition of Rhizoctonia solani extracts. The cumulative production of solavetivone was nearly doubled by integrated extraction of the products from the media during the 24 h accumulation period. Western blots with monoclonal antibodies against SC show that the enzyme levels are the same for both extracted and non-extracted cultures. SC activities measured in vitro with radioactive substrate are not significantly different. These results suggest that productivity is controlled by substrate availability within the terpenoid pathway, and feedback regulation precedes the branch-point enzyme sesquiterpene cyclase.

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Effects of compounds related to signal transduction on anthraquinone biosynthesis in transformed cells of Rubia cordifolia var. pratensis (형질전환시킨 갈퀴꼭두서니 세포의 색소생합성에 미치는 수종의 신호전달 cascade 관련물질의 효과)

  • Lyu, Lee;Shin, Seung-Won
    • Korean Journal of Pharmacognosy
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    • v.31 no.2
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    • pp.235-239
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    • 2000
  • The effects of several compounds related to signal transduction cascade were determined to induce the production of alizarin and purpurin in the hairy root culture system of Rubia cordifolia var. pratensis. It was found that out of five tested compounds jasmonic acid(1 mg/l) and methyl jasmonate(1 mg/l) stimulated strongly the biosynthesis of the pigments while linolenic acid(1 mg/l) induced no significant increase of the product. Yeast extract(600 mg/l) and arachidonic acid(1 mg/l) showed relatively mild inducing effects on production of alizarin. The effects of jasmonic acid and methyl jasmonate were reduced by treatment with cycloheximide(2.8 mg/l).

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Induction of Hariy Root and Bioreactor Culture of Lycium chinense (구기자나무 (Lycium chinense)의 모상근 유도 및 생물반응기 배양)

  • Bae, Ki-Hwa;Kim, Yun-Soo;Jeong, Jae-Hun;Kim, Young-Seon;Choi, Yong-Eui;Yoon, Eui-Soo
    • Journal of Plant Biotechnology
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    • v.31 no.4
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    • pp.295-300
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    • 2004
  • This article was conducted to induce the transgenic hairy roots and determine the effect of culture conditions on optimum growth of hairy roots by Agrobacterium rhizogenes strain, 15834 in Lycium chinense Miller. Hairy roots of L. chinense Miller. were induced from leaf segments by co-cultivation with A. rhizogenes. When the hairy roots were cultured in various MS medium strength and sucrose concentrations, the highest growth of hairy roots was observed in half-strength MS media supplemented with 3% sucrose, respectively. In air lift bioreactor cultures, the liquid medium contained with 1/2 MS and 3% sucrose was also the best for optimum growth of hairy roots.

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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Effects of IAA, IBA, and Media on Growth and Saikosaponin Biosynthesis in Bupleurum falcatum Hairy Root Culture (시호 모상근 배양에서 생장과 시호 사포닌 생합성에 미치는 배지와 IAA 및 IBA의 영향)

  • Ahn, Jun Cheul;Kim, Eung Sik;Lee, Hyun Jin;Hwang, Baik
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.3
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    • pp.171-175
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    • 1999
  • We examined the effects of various basal media and exogenous auxins on the growth and saikosaponin production in hairy root cultures of Bupleurum falcatum. Of the various media tested, 2RCM medium was superior for growth and saikosaponin content, in which saikosaponin a, c, and d was produced, however MS medium was resulted in extremely low level in saikosaponin production. By the treatment of 0.5 mg/L IBA the growth rate was increased by about 60% in 2RCM medium, whereas both IAA and IBA showed a potent inhibition effect in saikosaponin biosynthesis by the treatment of 0.01 mg/L~5mg/L.

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Physiological Studies on the Formation of Hairy Root by the A. rhizogenes. III. Attachment of A. rhizogenes strain A4 to Carrot(Daucus carota L.) Cells (Agrobacterium rhizogense에 Hairy Root 형성에 대한 생리학적 연구. III. 당근 세포에의 A. rhizogenes의 부착)

  • Hwang, B.;Hwang, S. J.;Ann, J. C.;Jo, H. S.
    • KSBB Journal
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    • v.4 no.2
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    • pp.94-98
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    • 1989
  • In vitro attachment experiments of bacteria to surface of host plant cell were carried out using C14 labeled cells of A. rhizogenes strain A4 and carrot protoplasts isolated from suspension culture of cells. Protoplasts were cocultivated with A. rhizogenes at various times after their isolation. Attachment kinetics showed that adherence of bacteria to protoplasts attained a maximum level within 120mins of co-cultivation. Maximum attachment occured at pH 6.0 and 24-35$^{\circ}C$. Bacterial attachment was observed at botg carrot cells with and without primary cell wall. The inhibition of transformation on the carrot root discs by A. rhizogenes was observed when non-related strain and heat inactivated bacterial strain cells were pretreated.

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The Effect of NaCI on the Growth and Ginsenoside Production from Ginseng Hairy Root (인삼모상근의 생장과 Ginsenoside 생산에 미치는 NaCl의 영향)

  • Kim, Yu-Jin;Sim, Ju-Sun;;Lee, Chung-Hyae;In, Jun-Gyo;Lee, Bum-Soo;Yang, Deok-Chun
    • Korean Journal of Medicinal Crop Science
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    • v.16 no.2
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    • pp.94-99
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    • 2008
  • Korean ginseng (Panax ginseng C.A. Meyer) is very difficult to obtain stable production of qualified ginseng roots because of variable stresses in soil environments. High salt concentrations in the ginseng nursery soil environment of Korea is one of important reducing factors for the stable production of quality ginseng. These studies were accomplished to identify the growth rate and production of ginsenoside from ginseng hairy root against NaCI. In the MS liquid culture, the highest contents and productivity of ginsenosides were appeared at 4 week after onset of the treatment of 0.1 M NaCI. And 0.24 M NaCI was more effective on the growth of ginseng hairy root under light condition than dark condition. Plants generally produce secondary metabolites in nature as a defense mechanism against pathogenic and insect attack. In this study, NaCI acts as a kind of stress as well as elicitor for production of ginsenosides.

Protoplast Fusion of Nicotiana glauca and Solanum tuberosum Using Selectable Marker Genes (표식유전자를 이용한 담배와 감자의 원형질체 융합)

  • Park, Tae-Eun;Chung, Hae-Joun
    • The Journal of Natural Sciences
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    • v.4
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    • pp.103-142
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    • 1991
  • These studies were carried out to select somatic hybrid using selectable marker genes of Nicotiana glauca transformed by NPTII gene and Solanum tuberosum transformed by T- DNA, and to study characteristics of transformant. The results are summarized as follows. 1. Crown gall tumors and hairy roots were formed on potato tuber disc infected by A. tumefaciens Ach5 and A. rhizogenes ATCC15834. These tumors and roots could be grown on the phytohormone free media. 2. Callus formation from hairy root was prompted on the medium containing 2, 4 D 2mg/I with casein hydrolysate lg/l. 3. The survival ratio of crown gall tumor callus derived from potato increased on the medium containing the activated charcoal 0. 5-2. 0mg/I because of the preventions on the other hand, hairy roots were necrosis on the same medium. 4. Callus derived from hairy root were excellently grown for a short time by suspension culture on liquid medium containing 2, 4-D 2mg/I and casein hydrolysate lg/l. 5. The binary vector pGA643 was mobilized from E. coli MC1000 into wild type Agrobacteriurn tumefaciens Ach5, A. tumefaciens $A_4T$ and disarmed A. tuniefaciens LBA4404 using a triparental mating method with E. ccli HB1O1/pRK2013. Transconjugants were obtained on the minimal media containing tetracycline and kanamycin. pGA643 vectors were confirmed by electrophoresis on 0.7% agarose gel. 6. Kanamycin resistant calli were selected on the media supplemented with 2, 4-D 0.5mg/1 and kanamycin $100\mug$/ml after co- cultivating with tobacco stem explants and A. tumefaciens LBA4404/pGA643, and selected calli propagated on the same medium. 7. The multiple shoots were regenerated from kanamycin resistant calli on the MS medium containing BA 2mg/l. 8. Leaf segments of transformed shoot were able to grow vigorusly on the medium supplemented with high concentration of kanamycin $1000\mug$/ml. 9. Kanamycin resistant shoots were rooting and elongated on medium containing kanamycin $100\mug$/ml, but normal shoot were not. 10. For the production of protoplast from potato calli transformed by T-DNA and mesophyll tissue transformed by NPTII gene, the former was isolated in the enzyme mixture of 2.0% celluase Onozuka R-10, 1.0% dricelase, 1.0% macerozyme. and 0.5M mannitol, the latter was isolated in the enzyme mixture 1.0% Celluase Onozuka R-10, 0.3% macerozyme, and 0.7M mannitol. 11. The optimal concentrationn of mannitol in the enzyme mixture for high protoplast yield was 0.8M at both transformed tobacco mesophyll and potato callus. The viabilities of protoplast were shown above 90%, respectively. 12. Both tobacco mesophyll and potato callus protoplasts were fused by using PEG solution. Cell walls were regenerated on hormone free media supplemented with kanamycin after 5 days, and colonies were observed after 4 weeks culture.

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