• KSBB Journal
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• v.4 no.3
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• pp.288-292
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• 1989

Induction and culture of hairy roots from ginseng(Panax ginseng C. A. Meyer) roots discs by A. rhizogenes strain $A_4$ were studied. After 6-12 weeks infected with A. rhizogenes tumor and hairy roots emerged from the root discs. The ratio of hairy root induction on root discs was higher in 5-year old than in 3, 4, and 6-year old ginseng. On treatment with IAA, IBA, 2, 4-D and tryptophan, hairy roots formation showed a significant increase at 15-30mg/1 tryptophan treated. Subsequently, hairy roots were cultured on hormone-free RCM medium(pH 4.5, sucrose 30g/1).

• 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.

• Korean Journal of Pharmacognosy
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• v.27 no.4
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• pp.301-308
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• 1996

Hairy roots induced from stems of Rubia cordifolia var. pratensis were cultured in the liquid medium under a variety of auxins to find the optimal condition for the growth and production of pigments. Culture of the hairy roots on NN liquid medium containing NAA 0.5 mg/l was best for growth of hairy roots. Production of yellow anthraquinone derivatives and purpurin in hairy roots was enhanced by the culture on NN liquid medium without auxins. Effects of L-phenylalanine, L-tyrosine and juglone, synthesized via the shikimic acid pathway, on growth and production of pigments in hairy roots were studied in the present study. Concentration of exogeneous L-phenylalanine. L-tyrosine and juglone in liquid culture system of hairy root containing NAA 0.1 mg/l was decreased quickly in its early stages of the culture period. Addition of juglone to NN liquid medium containing NAA 0.1 mg/l enhanced the productivity of pigments in hairy roots.

• Plant Biotechnology Reports
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• v.5 no.3
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• pp.255-263
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• 2011

Codonopsis lanceolata (Campanulaceae) has been used in traditional medicines, as its roots contain several kinds of triterpenoid saponin with high medicinal values. In this work, we induced transgenic hairy roots of C. lanceolata and analyzed triterpenoid saponins from the hairy roots and hairy root-derived transgenic plants. Hairy roots were obtained from leaf explants by the transformation of Agrobacterium rhizogenes R1000. Transgenic hairy root lines were confirmed by the transcriptional activities of rolA, B, C, and D genes by RT-PCR. Transgenic root lines actively proliferated on hormone-free medium but not in nontransformed roots. Hairy roots contained richer triterpenoids (lancemaside A, foetidissimoside A, and aster saponin Hb) than nontransformed roots. Transgenic plants were regenerated from the hairy roots via somatic embryogenesis. They showed phenotypic alterations such as shortened shoots and an increased number of axillary buds and adventitious roots. The transgenic plants also contained higher triterpenoid levels than wild-type plants. These results suggest that hairy roots and transgenic plants of C. lanceolata could be used as medicinal materials for the production of triterpene saponins.

• Korean Journal of Plant Tissue Culture
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• v.25 no.6
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• pp.525-530
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• 1998

These studies were carried out to select the active grow hairy root lines induced from various ginseng(Panax ginseng C. A. Meyer) parts. Hairy roots were induced in root explants, stem and petiole in vitro by A. rhizogenes R1000 or A. rhizogenes $A_4$. These hairy roots could be grown on the phytohormone free medium, and PCR analysis of rol C and vir C gene fragments confirmed that hairy roots were transgenic tissues. We have selected 11 hairy root lines with active growing characters among 300 hairy root lines selected based on growth and morphological characteristics on 1/2MS solid media with 250 mg/L carbenicillin. Morphological characteristics of selected 11 hairy root lines were thickness and thiness of main roots, and many projection for lateral roots, active grow of lateral roots. Among selected 11 hair root lines prominent characteristics of hairy roots with active growing characters were thiness of main roots and active grow of lateral roots. But characteristics of low growing hairy roots were thickness of main roots and low grow of lateral roots. Finally we have selected actively growing hairy roots, KGHR-1, KGHR-5, KGHR-8 among 11 hairy root lines.

• Journal of Ginseng Research
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• v.20 no.3
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• pp.318-324
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• 1996

The effects of light on the growth and ginsenosides production were examined in the hairy roots of Panax ginsen C.A. Meyer induced by Agrobacterium rhizogines A4. The 9rowth of ginseng hairy roots in 1/2MS liquid medium was significantly decreased with an increment of light intensity (1,000~7,000 lux). The growth of hairy roots under 7,000 lux condition was decreased at 17% compared to the dark condition. The production of 7 ginsenosides in hairy root was very high in 3,500 lux condition. The production of ginsenoside-Rg, and Rf increased 3.3 and, 2.4 times respectively as compared to dark condition. The growth of hairy roots was inhibited by blue light, while ginsenosides production was increased. The sucrose demands of hairy roots was examined in light condition(3,500 lux). The growth of hairy roots in 1/2MS liquid medium with various sucrose concentrations(1~4%) was high in IVp sucrose, while ginsenosides production was high in 3% sucrose condition. The growth and ginsenosides production were high when hairy roots were cultured in dark condition for 1 week and then transferred to light condition(3,500 lux) for 4 weeks. It is suggested that ginsenosides production could be accelerated by light intensity of specific wavelength in cultures of ginseng hairy roots.

• Journal of Photoscience
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• v.8 no.1
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• pp.19-22
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• 2001

It was reported that Red ginseng contains specific ginsenoside-Rg$_2$,-Rg$_3$,-Rh$_1$and -Rh$_2$, which show various pharmacological effects. However, production of these specific ginsenosides from Red ginseng is not commercially applicable because of high cost of the raw material, roots. This work was carried out to examine the production of Red ginseng specific ginsenosides from Agrobacterium-transformed hairy roots. Hairy roots were induced from 3 year-old root segment of Korean ginseng (Panax ginseng C.A. Meyer) after infection with Agrobacterium rhizogenes A4. Among many lines of hairybroots, KGHR-8A was selected. Steam heat treatment of hairy roots was resulted in the changes of ginsenoside composition. Eleven ginsenosides were detected in heat-treated hairy roots but eight in freeze dried hairy roots. In heat treated hairy root, content of ginsenoside-Rb$_1$,Rb$_2$,Rc, Rd, Re, Rf, and Rg$_1$were decreased compared to those of freeze dried hairy roots. However, heat treatment strongly enhanced the amount of Red ginseng specific ginsenogides (ginsenoside-Rg$_2$,-Rg$_3$,-Rh$_1$and -Rh$_2$). Amounts of ginsenoside-Rg$_3$,-Rh$_1$and -Rh$_2$ in heat-treated hairy roots were 2.58, 3.62 and 1.08 mg/g dry wt, respectively, but these were detected as trace amount in hairy roots without heat treatment. Optimum condition of heat treatment for the production of Red ginseng specific ginsenoside was 2 h at 105$^{\circ}C$. This result represents that Red ginseng specific ginsenoside can be producted from hairy roots by steam heat treatment.

• Journal of Plant Biology
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• v.33 no.3
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• pp.183-188
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• 1990

Induction and culture of hairy roots by Agrobacterium rhizogenes A4 were carried out in Platycodon grandiflorum DC. After 2-4 weeks of inoculation with Agrobacterium rhizogenes hairy roots were formed at root segments in the balloon flower. Optimized growth of hairy roots was obtained in hormone-free MS medium, 6% sucrose and pH 5.8. The pattern of ginsenoside in the transformed roots was not different with that in the ordinary roots.

• 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.

• Molecules and Cells
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• v.22 no.3
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• pp.269-274
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• 2006

In order to characterize saikosaponin biosynthesis in Bupleurum falcatum, the expression of five isoprenoid pathway genes and their relationship to saikosaponin accumulation in the hairy roots were analyzed. The hairy roots exhibited a rapid accumulation of saikosaponins when incubated in a root culture medium (3XRCM). Homology-based RT-PCR was used to isolate core fragments of five genes, HMGR, IPPI, FPS, SS, and OSC, from the hairy roots. The deduced amino acid sequences exhibited amino acid identities of more than 85% to previously reported genes. Using the fragments as probes, the expression of these five genes in the hairy roots during incubation in 3XRCM medium was examined. Expression of all five genes in the hairy roots increased soon after incubation. In particular, the SS and OSC genes were coordinately induced at 8 days of incubation, and their expression persisted throughout the incubation period. A quantitative HPLC analysis showed that the saikosaponin content of the hairy root culture also began to increase at 8 days of culture. The correlation between SS transcript level and saikosaponin content in the hairy roots suggests that transcriptional regulation plays a regulatory role in saikosaponin biosynthesis.