• Title/Summary/Keyword: transformed root cultures

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Detection of Lignans from Transformed Root Cultures of Schisandra chinensis Baillon (오미자의 형질전환된 근으로부터 리그난 화합물의 검출)

  • Hwang, Sung-Jin;Pyo, Byoung-Sik;Hwang, Baik
    • Korean Journal of Medicinal Crop Science
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    • v.12 no.6
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    • pp.448-453
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    • 2004
  • Transformed roots of Schisandra chinensis were obtained following co-cultivation of in vitro cultivated plantlet segments with Agrobaterium rhizogens ATCC15834. This root was examined for its growth and gomisin J contents under various culture conditions. Among the six basal culture media tested, WPM (Lloyd & McCown, 1980) medium supplemented with 5% sucrose was the best roots growth 6.2 (g D.W/flask) and gomisin J accumulation 1.56 $(X10^{-3}\;ug/g\;D.W)$. Initial inoculum size correlated with the yield of biomass while gomisin J contents was not affect. Gomisin J production was influenced by the initial sucrose concentration and the highest production yield was achieved at the concentration of 7%. The optimal shaking speeds for roots growth and gomisin J production was 120 and 140 rpm, respectively.

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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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.

Production of Saponin by Hairy Root Cultures of Ginseng (Panax ginseng C.A. Meyer) Transformed with Agrobacterium rhizogenes (Agrobacterium rhizogenes에 의하여 형질전환된 인삼(Panax ginseng C.A. Meyer)의 모상근 배양에 의한 Saponin 생산)

  • Hwang, Baik;Ko, Kyeong-Min;Hwang, Kyeong-Hwa;Hwang, Sung-Jin;Kang, Young-Hee
    • Journal of Plant Biology
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    • v.34 no.4
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    • pp.289-296
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    • 1991
  • Cultures of hairy root induced from ginseng(Panax C.A. Meyer) transformed with Agrobacterium rhizogenes (strain A4, ATCC 15834) were established and morphologically two different hairy root strains (HB1, HB2) were obtained. To determine the optimum growth rate, the hairy root (HB2) was cultured in various liquid medium supplemented with or without plant growth hormone. The growth rate of hairy root cultured on MS medium was 1.3-3.1 times higher than those cultured on other media, and the optimum sucrose concentration and pH were 3-6%, 5.5-6.5, respectively. Also, the growth rate of hairy root was increased when 0.02 M ammonium nitrate, 1.2 mM potassium phosphate (monobasic) and 0.5 mg/l IBA were supplied to liquid medium. The saponin patterns and contents of hairy root (HB2) were determined by TLC and HPLC. The crude saponin contents were 4.67% and the total saponin contents were 1.0%, on dry weight basis.

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Cytogenetic and Histological Characteristics of Ginseng Hairy Root Transformed by Agrobacterium rhizogenes (Agrobacterium rhizogenes에 의하여 형질전환된 인삼 모상근의 세포 유전학적 및 조직학적 특성)

  • 고경민
    • Journal of Plant Biology
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    • v.36 no.1
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    • pp.75-81
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    • 1993
  • Ginseng (Panax ginseng C.A. Meyer) hairy root transformed with Agrobacterium rhizogenes (strain A4) was examined cytogenetically and histologically to assess its characteristics. The optimum growth of hairy root obtained in hormone-free MS medium (sucrose 30 g/L, pH5-6) for long period cultures. All hairy root strains (HB1, HB2, HB3) had the 2n diploid number (2n=48) of chromosomes in root tip cells. There were no alteration in chromosome structure except in one cell of HB3 strain. Results of SDS-PAGE showed a few difference in pattern and number of bands between normal and hairy root of ginseng. The root anatomy of normal root and hairy root differed each other. The hairy root had a clearly defined vascular strand, and the morphology of cortical cells were disorganised with large intercellular spaces.

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Rutin production in hairy root cultures of buckwheat(Fagopyrum esculentum Moench)

  • Park, Sang-Un;Kim, Yeon-Bok;Park, Cheol-Ho
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2002.11b
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    • pp.23-23
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    • 2002
  • We established hairy root cultures of F. esculentum transformed with A. rhizogenes for in vitro rutin production. Additionally, we describe the effects of different media and plant growth regulators on growth and rutin biosynthesis in buckwheat hairy root cultures. Excised leaves of P. tinctorium from 10-day-old seedlings were used as the explant material for co-cultivation with A. rhizogenes 15834. The hairy culture of Fagopyrum esculentum Moench. was established by infecting leaf explants with Agrobacterium rhizogenes 15834. About four to five weeks after co-cultivation with A. rhizogenes, 10 hairy roots were excised from the necrotic explant tissues. After repeated transfer to fresh medium for three months, ten clones were transferred to MS liquid culture medium. The growth and rutin production of each clone differently response to the MS liquid medium. Among these clones, H8, which had exhibited good growth rate and one of the highest rutin productivity, was selected for the following experimment.(중략)

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Multiplication and Transformation of Medicinal Plants for Production of Useful Secondary Metabolites II. Establishment of Hairy Root Cultures of Centella asiatica

  • Paek, Yun-Woong;Hwang, Sung-Jin;Park, Don-Hee;Hwang, Baik
    • Journal of Plant Biology
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    • v.39 no.3
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    • pp.161-166
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    • 1996
  • The hairy root cultures of Centella asiatica were established by infection leaf explants with Agrobacterium rhizogenes A4, 15834 in 1/2 Murashing and skoog liquid medium supplemented with 50 $\mu$M acetosyringone. The induced hairy roots were subjected to paper electrophoresis for the detection of opine and opine-positive clones which were considered to have been transformed. Five hairy root clones were selected according to the different bacterial strains used, growth rate and pattern. Among media tested, MS basal medium substituted phosphate concentration by 2.5mM K2HPO4 showed the highest growth rate in the dark condition.

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Ginsenoside Production by Hairy Root Cultures of Panax ginseng Transformed With Agrobacterium rhizogenes (Agrobacterium rhisogense에 의해 형질 전환된 인삼의 모상근 배양에서 Ginsenoside의 생산)

  • 고경수;허인옥고정삼이윤진
    • KSBB Journal
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    • v.5 no.3
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    • pp.263-268
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    • 1990
  • New methods have been developed to transform Panax ginseng with Ri plasmids of Agrobacterium rhizogenes 15834 and A. rhizogenes A4. Modified leaf disc method was made feasible to establish hairy root culture even when an axonic plantlet was not available as in the case of P. ginseng. The contents of ginsenosides (Rgl, Rf, Rc, Rbl, and Rb2) in hairy roots. were determined by HPLC. Hairy root cultures, established as liquid culture in MS medium, was produced 0.34~1.19% ginsenosides on dry weight basis, and this result is significantly higher level than that of normal P. ginseng.

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Comparative Evaluation of Modified Bioreactors for Enhancement of Growth and Secondary Metabolite Biosynthesis Using Panax ginseng Hairy Roots

  • Jeong, Gwi-Taek;Park, Don-Hee
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.6
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    • pp.528-534
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    • 2005
  • Hairy root cultures have demonstrated great promise in terms of their biosynthetic capability toward the production of secondary metabolites, but continue to constitute a major challenge with regard to large-scale cultures. In order to assess the possibility of conducting mass production of biomass, and the extraction of useful metabolites from Panax ginseng. P. ginseng hairy roots, transformed by Rhizobium rhizogenes KCTC 2744, were used in bioreactors of different types and sizes. The most effective mass production of hairy roots was achieved in several differently Sized air bubble bioreactors compared to all other bioreactor types. Hairy root growth was enhanced by aeration, and the production increased with increasing aeration rate in a 1 L bioreactor culture. It was determined that the hairy root growth rate could be substantially enhanced by increases in the aeration rate upto 0.5vvm, but at aeration rates above 0.5vvm, only slight promotions in growth rates were observed. In 20 L air bubble bioreactors, with a variety of inoculum sizes, the hairy roots exhibited the most robust growth rates with an inoculum size of 0.1% (w/v), within the range 0.1 to 0.7% (w/v). The specific growth rates of the hairy root decreased with increases in the inoculum size.