• Title/Summary/Keyword: transformed hairy roots

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Culture of Hairy Roots Induced by Agrobacterium rhizogenes in Platycodon grandiflorum DC. (Balloon Flower) (Agrobacterium rhizogenes 에 의하여 유도된 도라지 (Platycodon grandiflorum DC.) Hairy Root 의 배양)

  • 김병노
    • Journal of Plant Biology
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    • v.33 no.3
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    • pp.183-188
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    • 1990
  • Induction and culture of hairy roots by Agrobacterium rhizogenes A4 were carried out in Platycodon grandiflorum DC. After 2-4 weeks of inoculation with Agrobacterium rhizogenes hairy roots were formed at root segments in the balloon flower. Optimized growth of hairy roots was obtained in hormone-free MS medium, 6% sucrose and pH 5.8. The pattern of ginsenoside in the transformed roots was not different with that in the ordinary roots.

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Production of Red Ginseng Specific Ginsenosides $(Rg_2, Rg_3, Rh_1 and Rh_2)$ from Agrobacterium-transformed hairy Roots of Panax ginseng by Heat Treatment

  • Yang, Deok-Chun;Yang, Kye-Jin;Park, Yong-Eui
    • Journal of Photoscience
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    • v.8 no.1
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    • pp.19-22
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    • 2001
  • It was reported that Red ginseng contains specific ginsenoside-Rg$_2$,-Rg$_3$,-Rh$_1$and -Rh$_2$, which show various pharmacological effects. However, production of these specific ginsenosides from Red ginseng is not commercially applicable because of high cost of the raw material, roots. This work was carried out to examine the production of Red ginseng specific ginsenosides from Agrobacterium-transformed hairy roots. Hairy roots were induced from 3 year-old root segment of Korean ginseng (Panax ginseng C.A. Meyer) after infection with Agrobacterium rhizogenes A4. Among many lines of hairybroots, KGHR-8A was selected. Steam heat treatment of hairy roots was resulted in the changes of ginsenoside composition. Eleven ginsenosides were detected in heat-treated hairy roots but eight in freeze dried hairy roots. In heat treated hairy root, content of ginsenoside-Rb$_1$,Rb$_2$,Rc, Rd, Re, Rf, and Rg$_1$were decreased compared to those of freeze dried hairy roots. However, heat treatment strongly enhanced the amount of Red ginseng specific ginsenogides (ginsenoside-Rg$_2$,-Rg$_3$,-Rh$_1$and -Rh$_2$). Amounts of ginsenoside-Rg$_3$,-Rh$_1$and -Rh$_2$ in heat-treated hairy roots were 2.58, 3.62 and 1.08 mg/g dry wt, respectively, but these were detected as trace amount in hairy roots without heat treatment. Optimum condition of heat treatment for the production of Red ginseng specific ginsenoside was 2 h at 105$^{\circ}C$. This result represents that Red ginseng specific ginsenoside can be producted from hairy roots by steam heat treatment.

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Baicalin Production in Transformed Hairy Root Clones of Scutellaria baicalensis

  • Hwang, Sung-Jin
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.2
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    • pp.105-109
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    • 2006
  • A transformed hairy root clone of Scutellaria baicalensis was established following infection with Agrobacterium rhizogenes ATCC15834. Three root clones of S baicalensis were selected by growth habit and baicalin content. The most active strain-the SR-03 clone-was examined for its growth and baicalin content under various culture conditions. The root growth and baicalin content were maximized in a Schenk and Hildebrandt medium supplemented with 4 and 6% sucrose, respectively. The accumulation of baicalin in transformed hairy roots was enhanced through exposure to various elicitors. Elicitation was attained by the addition of methyl jasmonate, salicylic acid, and various concentrations of fungal cell wall elicitors to the medium. The accumulation of baicalin in the elicited cultures ranged from 10.5 to 18.3 mg/g dry weight of the roots, which was 1.5- to 3-fold the amount attained in controls.

Physiological Studies on the Formation of Hairy Root by the Agrobacterium rhizogenes ; IV. Culture of Hairy Root and Survey of the Culture Condition. (Agrobacterium rhizogenes 에 의한 hairy root 형성에 대한 생리학적 연구 ; IV. Hairy root 배양 및 배양 조건에 관한 조사)

  • Hwang, Baik;An, Jun-Cheul;lee, Jae-Hyuk
    • KSBB Journal
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    • v.4 no.3
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    • pp.246-253
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    • 1989
  • Hairy roots of carrot were induced by Agrobacterium rhizogenes $A_4$ strain within about 2-4 weeks after inoculated from root disc. Early axonic culture is established in RCM agar medium and following is in MS rigid medium. After 15 days culture, the hairy roots were vigrous growth in about 10 times of initial inoculum. Anthocyanin contents of hairy roots were more than of ordinary roots. 2, 4-D ($10^{-4}mg/ l$), sucrose (5%), nitrogen source (0.03M) contained medium was optimized to growth of hairy root and contents of anthocyanin. Phenotypic alterations of leaves are observed in transformed plants and determined the transformation of hairy roots and the transformed plants by opine assay.

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Plant Regeneration from Hairy Root of Rehmannia glutinosa Liboschitz Transformed by Agrobacterium rhizogenes (형질전환된 지황의 모상근으로부터 식물체의 재분화)

  • Hwang, Sung-Jin
    • Korean Journal of Medicinal Crop Science
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    • v.14 no.1
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    • pp.31-35
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    • 2006
  • A protocol for plant regeneration from hairy root of Rehmannia glutinosa transformed by Agrobacterium rhizogenes ATCC15834 has been developed. Transgenic shoots were regenerated from hairy roots within 6 weeks after culture on the SH medium supplemented with 0.5 mg/l BA. Shoots were rooted on plant growth regulator free SH medium successfully. The transformed plants, which were regenerated from hairy roots, had thiner roots with extensive lateral branches, wrinkled leaves, shorter node, and grew faster compared with non-transformed plants. The biomass of the transformed plant was 1.28 g (F.W) per plant, significantly higher than the non-transformed plant (0.54 g F.W). The catalpol content in the transformed plant (0.56%) was also higher than that of the non-transformed plants (0.43%).

High Yield Saponin Production by Mass Cultures of Ginseng Transformed Tissue I. Induction, Culture of Transformed Tissue and Selection of High-Saponin-Producing Clones in Ginseng (인삼 형질전환 조직의 다량배양에 의한 Saponin 고 생산 I. 인삼에서 형질전환 조직의 유도, 배양과 Saponin 고 생산능주 선발)

  • 이정석;고경민
    • KSBB Journal
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    • v.9 no.2
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    • pp.157-164
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    • 1994
  • Hairy root clones of Panax ginseng were established by selection of some hairy roots formed on the leaf, stem and root segments transformed with Agrobacterium rhizogenes strain $A_4$. The transformed roots grew well in MS medium under the dark condition. To confirm the transformation with Ri-T-DNA, dot blot hybridization and opine analysis were Performed. Among four hairy roots induced from different part of ginseng, the HB3 hairy roots were examined for selection of high-saponin-producing clones. Four clones isolated from HB3 hairy root cultures displayed various phenotypes characterized by growth and total saponin content. Maximum growth was obtained for cultures of HB3-10 clone and the content of total saponin was 0.55 wt%. However, higher amount of total saponin was obtained with HB3-2 clone cultures(0.74 wt%) in spite of lower growth. Dot blot hybridization confirmed the introduction of Ri-T-DNA in the plant genome. In the opine test, agropine and mannopine were detected from all hairy root clones.

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Production of Anthocyanin by Culture of Hairy Roots of Raphanus sativus cv. Chungpihongsim (청피홍심무우(Raphanus sativus cv. Chungpihongsim)의 모상근 배양에 의한 안토시아닌 생성)

  • 안준철
    • Journal of Plant Biology
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    • v.35 no.1
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    • pp.37-43
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    • 1992
  • The hairy root culture of Raphanus sativus cv. Chungpihongsim was established by transformation with Agrobacterium rhizogenes $A_4$. The transformed roots grew well in adjusted Murashige and Skoog medium to 1/2 basal salts, pH 5.2, 3% sucrose. Agropine and mannopine, opine synthesized in the transformed tissue were detected in the extract of hairy roots. When 2, 4-D and kinetin were added in culture medium of hairy roots, the synthesis of anthocyanin was induced with disorganization of hairy root. Especially, addition of $0.45\;\mu\textrm{M}$ 2, 4-D and $2.3\;\mu\textrm{M}$ kinetin showed the maximum synthesis of anthocyanin. Pattern of anthocyanin synthesized in transformed roots was somewhat different from that of ordinary roots. However, aglycone part of all anthocyanin was identified as pelargonidin. The content of total anthocyanin in this sample was tentatively calculated 0.49 mg/g fresh weight.weight.

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Optimal Culture Conditions for Transformed Root Growth and Trichosanthin Formation in Trichosanthes kirilowii Max. (하늘타리 형질전환근의 생장 및 Trichosanthin의 생합성을 위한 최적화)

  • Hwang, Sung-Jin;Na, Myung-Suk
    • Korean Journal of Medicinal Crop Science
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    • v.15 no.1
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    • pp.46-50
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    • 2007
  • Transformed hairy roots were induced from in vitro grown plantlets of Trichosanthes kirilowii by infection with Agrobacterium rhizogenes strain ATCC15834. Transformed hairy roots exhibited active growth with high branching of roots on plant growth regulators-free medium. Cloned line (TR-03) of hairy root was tested for its growth and extracellular protein accumulation in medium under various culture conditions. Among the culture media tested, a full-strength MS medium had a pronounced effect on root biomass and extracelluar protein accumulation in medium. The maximum root biomass (2.4 g DRW/flask) and extracellular total protein contents $(28.3ug/m\ell)$ in medium was obtained at inoculum size of 2 g (FRW) and in MS medium supplemented with 4% sucrose. In addition, the optimal shaking speed for root growth and extracellular protein accumulation in medium were 100 rpm. The total extracellualr protein concentration reached a maximum of $28.3ug/m\ell$ at 4 weeks and decreased thereafter. Protein translation inhibitory activity was observed in culture broths and reached levels of 21.3 unit. These studies demonstrate that the transformed hairy roots can be utilized for the in vitro production of ribosome-inactivating proteins.

형질전환된 인삼 모상근의 배양조건 규명

  • Jeong, Gwi-Taek;Ryu, Hwa-Won;Gang, Chun-Hyeong;Park, Don-Hui;Hwang, Baek
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.413-416
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    • 2000
  • Panax ginseng hairy roots, transformed by Agrobacterium rhizogenes KTCT 2744, was incubated flask, and investigated its optimal growth conditions. Transformed P. ginseng hairy roots showed active branching pattern and fast growth at free-hormone medium. Optimal conditions of hairy roots showed maximum growth at $23^{\circ}C$, pH 5.8, 1/2 MS medium, 3% sucrose. Effects of combined sugar showed that it had a better growth at only sucrose without glucose or fructose than with glucose or fructose. Also the case of sucrose with glucose had a better growth than with fructose.

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Basic Studies on Cultivation of Transformed Plant Tissue in Bioreactor (생물반응기에서 형질전환된 식물조직 배양을 위한 기초연구)

  • 박성화;박돈희
    • KSBB Journal
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    • v.5 no.3
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    • pp.207-214
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    • 1990
  • Growth properties of carrot hairy roots transformed by Agrobacterium rhizogenes were compared in flask and bioreactor. Oxygen transfer coefficient KLa was measured during the cultivation in bioreactor. In flask cultures, initially sucrose 30g/l was nearly exhausted after 20days. pH was dropped from initially 5.8 to 4.79 after 4 days, but it is stable after that time. Finally, after 28 days, hairy roots were grown about twelve times. In view of the results studied optimum conditions, hairy roots were maintained high growth rates in sucrose 50g/l, pH 5.8, total nitrogen 60mM. Also in bioreactor cultures, fixed stainless sieve in bottom and aerated 0.31 vvm, the results of cultivation by the use of sucrose 50g/l had grown about twenty-eight times and pH variations were liked in flask. As a results, growth rate of 1.756g fresh weight/day/g inoculum in bioreactor were higher about three times than 0.57g fresh weight/day/g inoculum in flask culture. KLa values were showed a tendency to decrease from 0.209 min-1 to 0.068 min-1.

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