• Journal of Ginseng Research
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• v.31 no.3
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• pp.154-158
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• 2007

In order to increase the content of germanium in ginseng adventitious roots, the effects of chelating agents on germanium content and root growth were investigated in the submerged cultures of ginseng adventitious roots. Chelating agents such as citric acid, oxalic acid, phosphoric acid, EDTA (Ethylenediamine tetraacetic acid) or EGTA (Ethylene glycol-bis $({\beta}-aminoethylether)-tetraacetic$ acid) were administrated in the submerged culture of ginseng root containing 50 ppm $GeO_2$. After 6 weeks of cultivation, fresh weight, germanium and saponin contents in the roots were analyzed. Among chelating agents, addition of 1.0mM phosphoric acid was found to be best for germanium accumulation. Under this condition, germanium content increased 1.4 times as compared to that of the control. The germanium content in the adventitious roots also increased with addition of EDTA or EGTA, while they inhibited the growth of ginseng adventitious root. Citric and oxalic acids were not effective for increasing germanium content in adventitious roots. As the results, it suggests that the phosphoric acid can be proved as the optimal agent for the enhancement of germanium accumulation in ginseng adventitious roots. These results can be served as a guideline for the mass production of ginseng adventitious roots containing germanium by large-scale production.

• Korean Journal of Plant Resources
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• v.22 no.5
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• pp.454-460
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• 2009

Phyllanthus urinaria was an important species in Korea and distributed in all around of Korea. The roots and stems of this plant have been used for natural medicine for the treatment of diabetes, the hepatitis B virus and disturbances of the kidney and urinary bladder. Production of adventitious roots in P. urinaria by in vitro cultures could be used as alternatives materials. Shoot and root segments from P. urinaria seedling were cultured on Murashige and Skoog (MS) medium supplemented with 3.0 mg/L IBA and 30 g/L sucrose. After 4 weeks of culture, the highest induction of adventitious roots was obtained from the shoot part. Frequency of adventitious root formation on medium with various kinds of auxins (IAA, NAA, 2,4-D, and IBA) and various concentrations of IBA (0, 0.1, 0.5, 1.0, 3.0, and 5.0 mg/L) was tested. The maximun induction of adventitious root was obtained on medium with 0.5 mg/L IBA. In liquid culture, growth of root was best on medium supplemented with 30 g/L sucrose. Adventitious roots were cultured in 5 L bioreactor containing 1/2 MS medium supplemented with 0.5 mg/L IBA and 30 g/L sucrose and mass-production of adventitious roots was successfully achieved. These results revealed the first attempt for the production of adventitious roots in P. urinaria.

• Journal of Plant Biotechnology
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• v.49 no.3
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• pp.207-212
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• 2022

The optimal culture conditions for root organogenesis from the callus of peonies (Paeonia lactiflora Pall.) were investigated. Root explants with vascular bundles were cultured in Murashige and Skoog (MS) medium combined with 0.5-4.0 mg/L auxins (indole acetic acid [IAA], naphthalene acetic acid [NAA], indolebutyric acid [IBA], and 2,4-dichlorophenoxyacetic acid [2,4-D]) and 0.0-2.0 mg/L cytokinins (kinetin, zeatin, and benzylaminopurine [BAP]) to induce callus formation. The callus was then cultured in MS medium combined with three concentrations (0.1, 0.5, and 1.0 mg/L) of IAA, NAA, IBA, kinetin, zeatin, and BAP in the dark for 6 weeks. Based on the results, the effects of dark and light conditions on the callus cultured in MS medium with combinations of 0.1-1.0 mg/L IBA and zeatin for 6 weeks were studied. Callus formation was most effective (>+++) in the medium with a combination of 1.0 mg/L NAA and 1.0 mg/L zeatin. A high number of long adventitious roots were observed in the mediums with 0.1 mg/L IBA (6.66 and 4.82 cm) and 0.5 mg/L zeatin (2.32 and 0.72 cm) among auxins and cytokinins, respectively. The highest number (14.06) of adventitious roots were formed from the callus cultured in light in the MS medium combined with 0.1 mg/L IBA and 0.5 mg/L zeatin. This same medium induced the formation of the longest adventitious root (5.45 cm) in the dark. Thus, optimization of in vitro culture conditions may be possible for the mass propagation of adventitious roots in peonies.

• Journal of Ginseng Research
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• v.35 no.3
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• pp.283-293
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• 2011

With the purpose of improving ginsenoside content in adventitious root cultures of Korean wild ginseng (Panax ginseng Meyer), the roots were treated with different dosages of ${\gamma}$-ray (5, 10, 25, 50, 75, 100, and 200 Gy). The growth of adventitious roots was inhibited at over 100 Gy. The irradiated adventitious roots showed significant variation in the morphological parameters and crude saponin content at 50 to100 Gy. Therefore, four mutant cell lines out of the propagation of 35 cell lines treated with 50 Gy and 100 Gy were selected on the basis of phenotypic morphology and crude saponin contents relative to the wild type control. The contents of 7 major ginsenosides ($Rg_1$, Re, $Rb_1$, $Rb_2$, Rc, Rf, and Rd) were determined for cell lines 1 and 3 from 100 Gy and lines 2 and 4 from 50 Gy treatments. Cell line 2 showed more secondary roots, longer length and superior growth rate than the root controls in flasks and bioreactors. Cell line 1 showed larger average diameter and the growth rate in the bioreactor was comparable with that of the control but greater in the flask cultured roots. Cell lines 1 and 2, especially the former, showed much more ginsenoside contents than the control in flasks and bioreactors. Therefore, we chose cell line 1 for further study of ginsenoside contents. The crude saponin content of line 1 in flask and bioreactor cultures increased by 1.4 and 1.8-fold, respectively, compared to the control. Total contents of 7 ginsenoside types ($Rg_1$, Re, $Rb_1$, $Rb_2$, Rc, Rf, and Rd) increased by 1.8 and 2.3-fold, respectively compared to the control. Crude saponin and ginsenoside contents in the bioreactor culture increased by about 1.4-fold compared to that the flask culture.

• Journal of Plant Biotechnology
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• v.32 no.1
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• pp.57-61
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• 2005

The influence of different growth regulators on biomass of adventitious roots and secondary metabolites accumulation in bioreactor cultures of Eleutherococcus koreanum was studied. The maximum growth of adventitious roots was observed in the presence of $2.0\;\cal{mg/L}$ IBA (6.7 g DW/L). However the productivity of eleutheroside E was not significantly different among IBA levels. High level of thidiazuron (TDZ) efficiently increased both root growth and secondary metabolites production. Especially when $0.1\;\cal{mg/L}$ TDZ was combined with $3.0\;\cal{mg/L}$ IBA, the highest eleutheroside E accumulation ($3,327{\mu}g/g $DW) was observed. When the same TDZ level was treated in combination with $5.0\;\cal{mg/L}$ IBA, both chlorogrnic acid and eleutheroside B were accumulated to maximum level. In contrast, zeatin remarkably suppressed the accumulation of both eleutheroside B and chlorogenic acid.

• Journal of Korean Society of Forest Science
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• v.98 no.2
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• pp.142-147
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• 2009

Hyoscyamus species is sources of the hypnotic and sedative drugs hyoscyamine and scopolamine. Single cells of Hyoscyamus niger were dissociated from suspension cultures and adventitious roots obtained from single-cell clones which were cultured on B5 medium containing 3% (w/v) sucrose, 0.1 mg/L IBA and 0.4% (w/v) gelrite. H. niger adventitious root lines showed wide variation in tropane alkaloids production and growth. An effective selection of 200 root lines was made possible by the application of the 'Dragendorff's reagent' for qualitative detection of the alkaloids from root. A high correlation coefficient (r=0.9390) was observed between the values obtained with the two methods based on HPLC and Dragendorff's reagent analysis. Among the selected roots, the highest scopolamine content was 16.64 mg/g DW (Hn-59), which was 8.82-fold more productive than the lowest alkaloid producing line (Hn-25). Here, we established an efficient selection method on tropane alkaloids production and suggest that the Dragendorff's reagent is of great practical value in selection of invisible compounds.

• Korean Journal of Medicinal Crop Science
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• v.12 no.5
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• pp.372-377
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• 2004

Scopolia parviflora Nakai, a rare and endangered species, is the sole plant producing tropane alkaloids (TA) among the Korean native species. In order to enhance TA productivity the SP72 root line was selected by screening 100 of root line, and the optimal culture media for root growth and TA production were investigated with the SP72 roots. Based on the several media, SH and 2B5 medium were determined as growth medium and White and NN medium as production medium. Among the four combinations of two-stage culture, 2BN (2B5 as growth medium plus NN as production medium) showed more enhanced root growth and TA production as compared with production media of White and NN medium and growth media of SH and 2B5 medium, respectively. However, bubble column bioreactor (BCB) cultures applying two-stage culture did not reveal the effective results despite of the each successful operation of two-stage culture in conical flasks and BCB cultures.

• Journal of Plant Biotechnology
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• v.32 no.2
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• pp.111-116
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• 2005

The two different ways to supply air inside the bioreactor were examined in the adventitious root cultures of Panax ginseng C.A. Meyer. First, the aeration rate varied at 0.05, 0.1, 0.2 and 0.3 vvm, respectively which were supplied during the whole culture period. Second, the amount of air supply was increased from 0.05 to 0.3 vvm at 10-day intervals in proportion to the root growth. Both the root growth and the ginsenoside accumulation were maximized to 175.8 g dry wt. of root growth and 4.3 mg/g dry wt. of ginsenoside accumulation when the aeration rate was increased gradually. The effect of the sparger pore size (15, 30 and $60\;{\mu}m$) in the bioreactor was also investigated, which suggested the greatest root growth (175.9 g dry wt.) in the $15{\mu}m$-sized sparger and the highest ginsenoside content (4.3 mg/g dry wt.) in the $60\;{\mu}m$ size. Finally, the diameter of a sparger ($15\;{\mu}m$-sized) varied at 1.5, 3.0, 5.0 and 8.0 cm, respectively. The highest root growth (191.9 g dry wt.) and the ginsenoside content (4.9 mg/g dry wt.) were obtained in the sparger diameter of 8.0 cm.

• Journal of Korean Society of Forest Science
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• v.98 no.4
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• pp.363-369
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• 2009

This study was carried out to investigate the effect of ${NO_3}^-$ and ${NH_4}^+$ on the adventitious root growth and eleuthroside synthesis of Eleutherococcus koreanum in 5 L-bioreactor culture. The change in the medium components was also measured during culture. The fresh weignt of adventitious root reached to the highest level of 30.8 g FW/L in the presence of both 50 mM ${NO_3}^-$ and 10 mM $NH_4^+$, representing 3.6-fold increase compared to the 60 mM ${NH_4}^+$ alone. However, as the increase of the portion of ${NH_4}^+$, the root growth was decreased. However, the maximum eleutheroside B, E and E1 contents were $57.3{\mu}g/g$ DW, $188.4{\mu}g/g$ DW and $47.3{\mu}g/g$ DW, with 30 mM, 60 mM and 15 mM total nitrogen source, respectively. Fresh weight of adventitious root increased up to 6.8-fold of inoculum size within 9 weeks. The amounts of ${NH_4}^+$, $K^+$, ${NO_3}^-$ and ${PO_4}^-$ were decreased during culture periods. Based on these results, we suggest that various further studies are required to increase the biomass and the useful secondary metabilites.

• Proceedings of the Korean Society of Plant Biotechnology Conference
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• 2005.04a
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• pp.178-178
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• 2005