• Title/Summary/Keyword: Hairy root

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Induction of Ginseng Hairy Roots And Their Possible Application To Large Scale Culture

  • Yang, Deok-Chun
    • Plant Resources
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    • v.6 no.1
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    • pp.1-6
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    • 2003
  • 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 hairy roots induced by introducing Ri-plasmid of Agrobacterium rhizogenes into genomic DNA of plant cells show vigorous growth, and the hairy roots produce the same or more saponins than natural ginseng roots. Therefore, hairy roots can be used for commercial purposes. The present study was carried out to induce hairy roots with both active growth and high saponin contents. Numerous hairy roots of Panax ginseng were obtained after root disks of three-year old roots were infected with Agrobacterium rhizogenes R1000 A4T in dark condition after one month of culture. About 3 hundred lines of hairy roots were selected according as morphological characters on medium with carbenicillin. After pre-selection of fifteen lines of hairy roots with active growth, KGHR-l and KGHR-8 lines were finally selected which had characters of high content of ginsenoside-Rd and ginsenoside-Re, respectively. 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 5L Erlenmeyer flasks, lL roller drums, 10L jar-fermenters, and especially in 20L air-lift culture vessels.

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Continuous Production of Natural Colorant, Betacyanin, by Beta vulgaris L. Hairy Root

  • Kim, Sun-Hee;Ahn, Sang-Wook;Bai, Dong-Kyu;Kim, Kwang-Soo;Hwang, Baik;Lee, Hyeon-Yong
    • Journal of Microbiology and Biotechnology
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    • v.9 no.6
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    • pp.716-721
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    • 1999
  • It has been known that continuous cultivation of hairy root is difficult to maintain for a long period of time compared to the microbial and callus cultures. Chemostat cultivation was successfully carried out in order to economically produce a plant-based colorant, betacyanin, from red beet hairy root for more than 85 days in a 14-1 fermentor. The result from the chemostat cultivation was compared to those of the batch and fed-batch cultivations of red beet hairy roots. It was shown that hairy root reached its steady state within 50 days of the cultivation, and then maintained for about 25-30 days in a wide range of dilution rates. Total betacyanin production from the continuous process was also calculated to be 2.65g at 0.28(l/d) of dilution rate, compared to 0.196g from fed-batch cultivation. It was found that betacyanin production was a partially growth related process, yielding 0.376 mg/g-fresh wt. cell and $1.89{\times}10^{-5}$ mg/g-fresh wt. cell/d, with 0.92 of correlation factor in a partial growth-product model. It was also shown that the cell growth required was relatively large for maintenance amount of energy at a low dilution rate. The growth of hairy root was inhibited by high light intensity in following a photo-inhibition model. The growth parameters were estimated to be 0.3(l/d), $10.56kcal/\textrm{m}^2/h$,{\;}and{\;}35.81kcal/\textrm{m}^2/h$ for the maximum specific growth rate, half saturation light intensity, and inhibition light intensity, respectively.

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Biosynthesis of Sesquiterpene in Hairy Root and Cell Suspension Cultures of Hyoscyamus muticus by Elicitation Using Rhizoctonia solani Extracts (Rhizoctonia solani 추출액 첨가에 의한 Hyoscyamus muticus의 현탁세포배양 및 모상근배양에서 Sesquiterpene 생합성)

  • 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.279-284
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    • 1997
  • The extracellular sesquiterpenoids were accumulated in cell and hairy root cultures of Hyoscyamus muticus by elicitation using extracts of Rhizoctonia solani. The vetispiradiene synthase (VS) which is the first committed step in biosynthetic pathway leading to formation of solavetivone, lubimin, and rishitin from isoprenoid intermediate farnesyl pyrophosphate was induced upon elicitation, whereas no sesquiterpenoids and VS activity were detected in both control cell and hairy root cultures. VS activity increased rapidly and reached its maximum 12 h in both cell and hairy root cultures upon elicitor treatment. VS activities were paralleled with the absolute levels of VS polypeptide(s). Interestingly, the profiles of sesquiterpenoid accumulation in hairy root cultures were different from those in cell cultures. The hairy root culture seemed to fail to metabolize solavetivone further to lubimin.

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Production and Identification of Anthocyanin in Hairy Root Cultures of Ginseng (인삼 모상근 배양에 의한 Anthocyanin의 생산과 동정)

  • Ko, Kyeong-Min;Choi, Yang-Soon;Hwang, Baik
    • Journal of Plant Biology
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    • v.37 no.1
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    • pp.85-91
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    • 1994
  • In hairy root cultures of ginseng (Panax ginseng C.A. Meyer) transfonned by Agrobacterium rhizogenes, the effects of light, carbon source and various honnone on hairy root growth and anthocyanin production were investigated. Anthocyanin synthesis began to first occur 5 days after exposure to light, and then maximum yield of anthocyanin was 0.36 mg/g(fr wt) in MS medium after 30 days. Of the nutritional factors concentration of 60 mM nitrogen and sucrose as a carbon source showed marked effects on the growth and anthocyanin productiom MS medium supplemented with 0.5 mg/L IAA was most suitable for the hairy root proliferation, and the best accumulation of anthocyanin was obtained at 1 mg/L IAA treatment (0.41 mg/g, fr wt). Whereas 2,4-D tended to restrain the pigment synthesis. From the isolation and identification of anthocyanin pigments, main anthocyanin in ginseng hairy root was identified as pelargonidin-glucoside.coside.

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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.

Antibody-Mediated Resistance to Rhizomania Disease in Sugar Beet Hairy Roots

  • Jafarzade, M.;Ramezani, M.;Hedayati, F.;Mokhtarzade, Z.;Zare, B.;Sabet, M.S.;Norouzi, P.;Malboobi, M.A.
    • The Plant Pathology Journal
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    • v.35 no.6
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    • pp.692-697
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    • 2019
  • Agrobacterium rhizogenes-mediated transformation of sugar beet hairy roots expressing single-chain variable fragment (scFv) was exploited to evaluate the efficacy of four antibody-based constructs for interfering with the Beet necrotic yellow vein virus infection. The scFv specific to a major coat protein of virus, p21, was targeted to various cellular compartments including the cytosol (pIC and pICC constructs), apoplast (pIA), and mitochondrion (pIM). After mechanical virus inoculation, most of the hairy root clones expressing scFv in the cytosol displayed low virus titers while the majority of transgenic hairy root clones accumulated antibody in outer membrane of mitochondria or apoplast were infected. This hairy root system provided an efficient and rapid approach to initially investigating root disease resistance like rhizomania prior to transform whole recalcitrant plants such as sugar beet.

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.

The Production of Ginsenosides from Ginseng Hairy Root by Treatment of the Chitin and Chitosan (Chitin과 Chitosan 처리에 의한 인삼모상근으로부터 Ginsenosides 생산)

  • 오승용;박효진;최경화;맹성주;양계진;양덕춘
    • Journal of Ginseng Research
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    • v.24 no.2
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    • pp.68-73
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    • 2000
  • To elucidate the effect of chitin and chitosan on the production of ginsenosides and growth, ginseng hairy root was cultured on the 1/2 MS medium supplemented with chitin and chitosan of various concentrations and culture period. The highest growth was obtained with 1 mg/L of chitin. However, the growth was inhibited by 20 mg/L or above. The contents and productivity of ginsenosides were the highest when ginseng hairy roots were cultured on 40 mg/L chitin and applied of the third-weeks of culture period. Ginseng hairy root culture with 1 mg/L of chitosan resulted in the best growth, but the highest ginsenosides level was appeared in 30 mg/L chitosan. Ginsenosides content was increased when it was treated at the forth-week after culture as 30 mg/L of chitosan.

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Optimization of SOD Biosynthesis by Controlling Sucrose Concentration in the Culture of Carrot Hairy Root

  • Kim, Ji-Hyeon;Yoo, Young-Je
    • Journal of Microbiology and Biotechnology
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    • v.12 no.4
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    • pp.617-621
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    • 2002
  • In order to optimize the carrot hairy root culture for SOD production, a fed-batch culture of hairy roots was performed in a bioreactor. Maximum SOD activity was obtained when the hairy roots were transferred to the MS medium containing 110 g/1 concentration of sucrose. By controlling the sucrose concentration (70 g/1 sucrose for growth and 110 g/1 sucrose far production, respectively) In a two-stage fed-batch culture, 29 g/1 of the hairy roots was obtained based on the final dry mass. The volumetrically determined SOD activity and productivity in the fed-batch culture were about 6 times higher than those from the flask culture containing sucrose at 30 g/1 concentration.

Transformation and Hairy Root Culture in Solanum nigrum by Agrobacterium rhizogenes (Agrobacterium rhizogenes를 이용한 까마중의 형질전환(形質轉換)과 모상근(毛狀根) 배양(培養))

  • Ko, Kyung-Soo;Heo, In-Ok;Yang, Kwan-Pal;Lee, Woon-Jin;Kim, Chang-Min;Jo, Pill-Hyeong
    • Korean Journal of Pharmacognosy
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    • v.22 no.1
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    • pp.26-32
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    • 1991
  • The study aimed to confirm transformation, morphology, steroid alkaloidal TLC pattern and growth rate of hairy roots. The Solanum nigrum plantlets were inoculated with Agrobacterium rhizogenes strain 15834. Hairy roots were induced by plasmid. Agropine and mannopine were detected in the hairy roots. The organization of hairy roots on the transactional morphology was undifferentiated. Culture on the medium containing hormone(IBA 2, kinetin 0.1mg/l) altered hairy roots into callus. The growth rate of hairy roots on the NN30 liquid medium was 52 times heavier than in the original state and this was higher than on the other media. The results of TLC analysis indicated that the hairy roots produced steroidal alkaloids resembling those of normal roots.

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