• Title/Summary/Keyword: hairy roots

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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.

Mass Culture and Ginsenoside Production of Ginseng Hairy Root by Two-Step Culture Process (2계단 배양방법을 이용한 인삼 모상근의 대량배양과 Ginsenoside 생산)

  • Ko, Kyeong-Min;Yang, Deok-Chun;Park, Ji-Chang;Choi, Kang-Ju;Choi, Kwang-Tae;Hwang, Baik
    • Journal of Plant Biology
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    • v.39 no.1
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    • pp.63-69
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    • 1996
  • A hairy root clone of Panax ginseng C.A. Meyer, HRB-15 was cultured iu various conditions with 3 L bubble type bioreactor to enhance both growth and ginsenoside production. The hairy roots were more rapidly grown under the dark condition than under the light condition. However, total amount of ginsenoside of hairy roots cultured under the light for 30 days increased 2 folds as compared with the dark condition and was 1.10% based on 6 ginsenosides. Especially, ginsenoside-Re was significantly increased and some ginsenosides except for ginsenoside-Re was slightly reduced. Also, the growth of hairy roots decreased about 30% as compared with the dark condition. In contrast, addition of sodium acetate led to decreased production of ginsenoside and growth of hairy roots under light condition. The influence of potassium dihydrogenphosphate concentration was examined in MS medium and a 1.25 mM concentration was found to be the most appropriate for growth and ginsenoside production under light condition. Two-step process of hairy roots culture with yeast elicitation or without ammonia in culture medium was developed to enhance growth and giusenoside synthesis. $50\;\mu\textrm{g}$ of yeast elicitor per g of fresh weight showed a synergistic effect on the ginsenoside synthesis of hairy roots on 20 days after culture. At that time, the content of total ginsenoside was 1.15%, while the growth of hairy roots decreased 21 % as compared with the dark condition. In addition, when elimination of ammonia on 20 days after culture, the content of total ginsenoside was 1.26% with significant increment of ginsenoside-Rd (0.27%) in addition to ginsenoside-Re and the growth of hairy roots decreased 10% as compared with the dark condition. In this system, we have demonstrated a unique two-step process of hairy root cultures to maximize biomass and secondary metabolites. It has found possibility to enhance ginsenosides production by growing hairy roots in this method.

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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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Mass Production of Gain-of-Function Mutants of Hairy Roots in Catharanthus roseus (일일초에서의 기능획득 돌연변이 모상근의 대량생산)

  • Ko, Suk-Min;Chung, Hwa-Jee;Lee, Hyo-Yeon
    • Korean Journal of Plant Resources
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    • v.24 no.5
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    • pp.514-520
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    • 2011
  • This study describes conditions for the mass production of mutant hairy root lines by co-cultivation with A. rhizogenes harboring the activation tagging vector pHC7. Various sources of explants were subjected to genetic transformation with A. rhizogenes to determine optimum conditions and cultivar for the highest frequency of hairy root formation on explants. Hairy root formation also were investigated in transformed hairy roots grown in various culture media. Finally, a total of approximately 2,500 lines of hairy root mutants were produced in this study. A managing system for metabolomics in hairy root lines also were established. These hairy root lines will be useful to determine functions of genes relating biosynthesis pathway of secondary metabolites.

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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Growth Pattern and Content of Tropane Alkaloids of Metabolic Engineered Scopolia parviflora Hairy Root Lines

  • Kang, Young-Min;Min, Ji-Yun;Kim, Won-Jung;Kim, Yong-Duck;Lee, Byung-Hynn;Choi, Myung-Suk
    • Korean Journal of Medicinal Crop Science
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    • v.12 no.2
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    • pp.123-128
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    • 2004
  • Hyoscyamine and scopolamine are two most common tropane alkaloids found in the Solanaceae. The pEB expression vector carrying Nspmt gene was transformed to Agrobacterium rhizogenes. The growth of transgenic hairy roots were approximately to 80% of the wild type root. Transgenic hairy roots are less developed on only their branch roots than those of the wild type root. The extracts from Nspmt transgenic hairy root lines, 3 and 5 contained between 3.52 and 4.23 mg/g dry weight as hyoscyamine and did between 5.23 to 6.40 mg/g dry weight as scopolamine. These results showed that the overexpression of the pmt gene enhanced tropane alkaloids production of S. parviflora transformed roots and this improvement affected both hyoscyamine and scopolamine production.

Hairy Root Culture of Daucus carota for Anthocyanin Production in a Fluidized-bed Bioreactor (유동층 생물반응기에서 anthocyanin 생산을 위한 당근의 모상근 배양)

  • Kim, C.H.;Lee, S.W.;Chung, I.S.
    • Applied Biological Chemistry
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    • v.37 no.4
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    • pp.237-242
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    • 1994
  • Hairy root culture of Daucus carota was investigated for anthocyanin production in a fluidized-bed bioreactor. The growth of hairy roots in this bioreactor increased 2.5 fold while anthocyanin production was lower. However, the anthocyanin production of hairy roots in a fluidized-bed bioreactor was enhanced 2.3 fold in response to the treatment of the fungal elicitor.

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Induction and Culture of High Polyacetylene-Yielding Hairy Roots in Ballon Flower (Pathycodon grandiflorum) (도라지(Platycodon grandirorum) 뿌리조직에서 고농도 함유 모상근의 유도 및 배양)

  • Hwang, Baik
    • Journal of Plant Biology
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    • v.38 no.4
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    • pp.337-341
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    • 1995
  • Hairy roots of Korean ballon flower (Platycodon grandiflorum A. DC) were induced from the root tissues infected with Agrobacterium rhizogenes ATCC 15834. Growth and polyacetylene [lobetyol (1), lobetyolin (2) and lobetyolinin (3)] production fo ten hairy root clones cultured in 1/4 Gamborg B5 (B5) liquid medium were determined. One selected hairy root clone (D6) grew well in hormone free-B5 liquid medium and showed maximum content of polyacetylenes at week 6 for 1 (0.375% dry wt) and at week 7 for 2 and 3 (3.030% and 0.206% dry wt, respectively) whose levels were much higher than those of the intact plant root (1:0.019%, 2:0.077% dry wt, 3 was not detected).

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Effects of Antioxidants on Growth and Betalain Production in Hairy Root Cultures of Phytolacca esculenta van Houtte (자리공(Phytolacca esculenta van Houtte) 모상근의 생장과 Betalain 생산에 미치는 항산화제의 효과)

  • 양덕조;김용해;최혜연;최철희;양덕춘
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.2
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    • pp.65-70
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    • 1995
  • The synthesis of betalain in hairy root cultures of Phytolacca esculenta van Houtte required the light, but the growth of hairy roots was inhibited 5 times under light condition(1500 1X) compared to dark condition. To investigate the growth inhibition of hairy roots under light condition, we surveyed the effects of several antioxidants on growth and betalain production if suspension cultures of hairy roots was increased 1.2-1.4 times under dark condition and 1.3-1.9 times under light condition by the treatment of ascorbic acid, glutathione, $\alpha$-tocopherol, sodium pyrosulfate and propylgallic acid. The betalain production was increased 1.2-2.1 times by antioxidants under light condition. The combination treatment of antioxidants didn't have any significant effect on the growth of hairy roots and betalain synthesis. The antioxidants of the betalain production were more effect under blue light than others. It was discussed that the endogenous oxidants may be produced under blue light.

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In vitro induction of hairy root from isoflavones-producing Korean wild arrowroot Pueraria lobata

  • Kim, Soo-Jung;Cha, Min-Seok;Lee, Eun-Ji;Kim, In-Hye;Kwon, Jung-Eun;Kang, Se-Chan;Park, Tae-Ho
    • Journal of Plant Biotechnology
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    • v.39 no.3
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    • pp.205-211
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    • 2012
  • Pueraria lobata is a perennial legume plant, widely distributed in the countries of East Asia. It is a medicinally important leguminous plant and produces various isoflavones such as puerarin, daidzein etc which have potential for preventing several chronic diseases including osteoporosis, cardiovascular disease and cancer. In this study, we tried to induce hairy roots in vitro from Korean wild arrowroot P. lobata and investigated the effects of hormones and light conditions. Initially leaf and stem segments were infected with Agrobacterium rhizogenes and incubated in different conditions. Hairy roots were induced from only stem segments and the induction was best at dark condition and the presence of IBA during incubation. Secondary roots were also significantly much more induced at the dark condition than at the 16 hours light condition. Among plant growth regulators of auxin, IBA was best for secondary root formation while 2,4-D, IAA and NAA produced callus or less hairy roots. The presence of the foreign gene rolC transferred by A. rhizogenes that plays a major role in hairy root induction was confirmed by PCR. The accumulation of isoflavones such as puerarin and daidzin was also confirmed. These results will facilitate mass production of hairy root and can be used for the production of functional substances from wild arrowroots.