• 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.

• 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.

• 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.

• 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.

• Plant Resources
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• v.6 no.3
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• pp.170-174
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• 2003

The important component for medical effect in ginseng is ginsenoside. Korea Ginseng & Tobacco Research Institute contains approximately 200 lines produced by inbred selection. It is assumed that ginseng lines containing high level of ginsenoside should be included in those lines. Besides, new breeding methods such as cell line selection in vitro and hairy root were recently developed. Therefore, this study was carried out to detect genes related to ginsenoside, and to use it for selection marker to select and distribute lines containing high level of ginsenoside. DNA was extracted from both ginseng roots and hairy roots, and the difference between the line containing high ginsenoside(KG101) and normal ginsenoside(KG103) were analysed. As a result, 28 out of 36 primers showed bands, and many primers showed band difference between ginseng lines. It is considered that the bands should be analysed using DNA sequence comparison to check if those are related to ginsenoside. In case of hairy roots of ginseng, almost no differences were found between two lines.

• Journal of Plant Biotechnology
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• v.48 no.2
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• pp.100-105
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• 2021

Genetic engineering is a potential approach to improve secondary metabolism in plants. In order to elucidate the effect of production of anthocyanin pigment 1 (PAP1) overexpression on the bioactivity of ginseng, we analyzed its antioxidant, antimicrobial, and anti-elastase activities in this study. Our results showed that PAP1 overexpression increased the production of polyphenolic compounds including anthocyanins. The antioxidant, antimicrobial, and anti-elastase activities were stronger in anthocyanin-overproducing ginseng hairy roots (AOX) than in wild ginseng hairy roots. Using a different solvent system (0, 30, 70, and 100% (v/v) EtOH), we revealed that variations in the contents of the polyphenolic compounds were highly correlated with changes in the antioxidant and antimicrobial activities of AOX. The antioxidant, antimicrobial, and anti-elastase effects of AOX highlight genetic engineering as a powerful approach to enhance the therapeutic properties of plants. Our results show that AOX could potentially have various functional applications in the cosmetic and pharmaceutical industries.

• Korean Journal of Plant Tissue Culture
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• v.27 no.1
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• pp.31-37
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• 2000

Proton extrusion by hairy roots of Panax ginseng C.A. Meyer was enhanced by iron-deficiency or single salt solutions of nitrate or ammonium salts. The enhancement effect of iron-deficiency was confined to the apical zones, whereas that of nitrate and ammonium was confined to the basal root zones. tinder iron-deficiency medium, the proton extrusion of ginseng hairy roots was higher at two times than that grown in the medium of iron-sufficiency. According to pH in the medium, uptake of inorganic elements showed various patterns for the kind of inorganic elements. $Ca^{2＋}$, $Mg^{2＋}$ and Fe$^{2＋}$ were highest uptake at the pH 4.8 and Mn$^{2＋}$, Zn$^{2＋}$ and Cu$^{2＋}$ were highest uptake at the pH 3.8. Free sugars and ginsenosides were analyzed by HPLC. Five kinds of free sugars were indentified in ginseng hairy roots. Glucose, fructose and sucrose were a major sugar in ginseng hairy roots and contents of free sugars were highest at pH 6.8 and pH 4.8, respectively. The contents of ginsenosides and phenolic compounds were highest at pH 5.8. This physiological responses in ginseng hairy root indicated that it may well be a good model system for the study of mechanism of mobilization and uptake of inorganic elements related with the red-colored phenomenon in ginseng roots.

• Proceedings of the Korean Society of Plant Biotechnology Conference
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• 2005.11a
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• pp.88-94
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• 2005

Plant tissue culture studies have been done for the preservation of medicinal plant resources and efficient production of pharmaceutically important secondary metabolites. Micropropagation methods for Cephaelis ipecacuanha have been established and these methods enabled much more efficient propagation of the plants than the conventional methods using seedling or layering. The C. ipecacuanha plants derived from tissue culture grew uniformly in the field and they showed higher alkaloid contents compared to the plants grown from seedlings. Hairy root cultures of C. ipecacuanha and Panax ginseng have been established by infection with Agrobacterium rhizogenes, and the production of important pharmaceuticals by these cultures have been successfully demonstrated. In the case of C. ipecacuanha, the highest alkaloid yields from the hairy roots cultured for 8 weeks were 2.75-fold cephaeline (5.5 mg) and one third emetine (0.7 mg) compared with those from the roots of one-year old plant propagated through shoot-tip culture and cultivated in a greenhouse (2.0 mg cephaeline and 2.0 mg emetine). In the case of P. ginseng, ginsenoside contents in the hairy roots optimally cultured for 4 weeks were much higher than those in the roots of 4-year old field-grown plant. Thus our medicinal plant tissue cultures demonstrate desirable properties. However, they are always exposed to danger of microbial contamination or unexpected trouble of culture facilities. Cryopreservation of plant tissue cultures is a reliable method for long-term preservation. Cryopreservation studies on these cultures are also presented.

• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.250-253
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• 2000

We researched effects of growth on initial inoculum size, liquid volume, and medium feeding rate etc. Cell suspension inoculated at low cell concentrations showed a typical growth reduction, whereas root cultures showed an improvement in growth. In this paper, Hairy roots showed high growth rate at 0.4 % inoculum size and 100 mL liquid volume in 250 mL flask cultures.

• 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.