• 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.

• Korean Journal of Plant Resources
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• v.18 no.2
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• pp.207-215
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• 2005

Korean ginseng(Panax ginseng C.A. Meyer) roots have long been known as the best medicinal plant and its pharmaceutical bio-activities have been proven by scientific analyses of their components - ginsenosides, acidic polysaccharides, phenolic compounds, fatty acids and organic acids etc. Ginseng hairy roots and callus have been cultured in vitro for stable supply of ginseng material. In this study, the amount of ginsenosides, fatty acids, acidic polysaccharides, phenolic compounds and organic acids in ginseng hairy roots and callus were compared. Higher amount of ginsenoside was found in ginseng hairy roots than ginseng callus. Higher amount of saturated fatty acid (palmitic acid) was found in callus and higher amount of unsaturated fatty acid (linoleic acid) was found in hairy roots. Acidic polysaccharide and phenolic compounds were contained by the same amount in both hairy roots and callus. Organic acids were found more in hairy roots.

• KSBB Journal
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• v.5 no.3
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• pp.263-268
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• 1990

New methods have been developed to transform Panax ginseng with Ri plasmids of Agrobacterium rhizogenes 15834 and A. rhizogenes A4. Modified leaf disc method was made feasible to establish hairy root culture even when an axonic plantlet was not available as in the case of P. ginseng. The contents of ginsenosides (Rgl, Rf, Rc, Rbl, and Rb2) in hairy roots. were determined by HPLC. Hairy root cultures, established as liquid culture in MS medium, was produced 0.34~1.19% ginsenosides on dry weight basis, and this result is significantly higher level than that of normal P. ginseng.

• 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.

• Journal of Plant Biotechnology
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• v.34 no.4
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• pp.285-291
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• 2007

This study describes conditions for the mass production of activation-tagged mutant hairy root lines of ginseng by cocultivation with Agrobacterium rhizogenes. Because it is not currently possible to produce progeny from transgenic ginseng, a loss-of-function approach for functional genomics cannot be appliable to this species. A gain-of-function approach is alternatively the choice and hairy root production by cocultivation of A. rhizogenes would be most practical to obtain a large number of mutants. Various sources of explants were subjected to genetic transformation with various strains of A. rhizogenes harboring the activation-tagging vector pKH01 to determine optimum conditions for the highest frequency of hairy root formation on explants. Petiole explants cocultivated with A. rhizogenes R1000 produced hairy roots at a frequency of 85.9% after 4 weeks of culture. Conditions for maximum growth or branching rate of hairy roots were also investigated by using various culture media. Petiole explants cultured on half strength Schenk and Hildebrandt medium produced vigorously growing branched roots at a rate of 2.6 after 4 weeks of culture. A total of 1,989 lines of hairy root mutants were established in this study. These hairy root lines will be useful to determine functions of genes for biosynthesis of ginsenosides.

• Korean Journal of Plant Resources
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• v.12 no.1
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• pp.1-9
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• 1999

The experiments were carried out to determine the optimum conditions for the induction of hairy roots in ginseng(Panax ginseng C.A. Meyer) by Agrobacterium spp. We were examined the antibiotics resistance of Agrobacterium spp and various ginseng parts, and the media for induction of hairy roots. The optimum concentration of NaOCl for sterilization of ginseng root segments without tissue damage with reduce of contamination was 7% NaOCl for 15-20 min and 9% NaOCl for 5 min, respectively. The more ginseng ages, the more contamination of ginseng root segment by sterilized in 7% NaOCl for 20 min, and especially in ginseng root segments with epidermis in six-year old roots. The growth of Agrobacterium spp were inhibited, but ginseng root segments was death in 30mg/L tetracycline. In 500mg/L cefotaxime or 500mg/L carbenicillin, the growth of Agrobacterium sup were inhibited, and root segments was grown normally. The optimum conditions for induction of hairy roots were using the root segments of three-year old ginseng cultured in 1/2MS medium supplemented with 500mg/L cefotaxime, and inoculation of Agrobacterium to root segments were better co-culture than smear method. After 2 weeks co-culture, the callus induced in cambium of root segments cultured in 1/2MS solid medium with 500mg/L cefotaxime. And then after 2 weeks, ginseng hairy roots were induced in callus of root segments. PCR analysis of rot C gene fragment confirmed that hairy roots were transgenic tissues.

• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.254-255
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• 2000

형질전환된 인삼 모상근의 여러 생물반응기에서의 대량배양 가능성을 알아보기 위하여 1/2 MS 배지를 기본배지(호르몬 무첨가, 3% sucrose)로 하여 실험을 수행하였다. 그 결과 플라스크 배양에 비하여 $2{\sim}3.5$배 높은 성장률을 보였으며, 그 성장이 왕성함을 알 수 있었다. 배양 후반기에 모상근의 뭉치 형성으로 물질전달의 어려움으로 인하여 성장에 장해를 미치는 것으로 사려된다.

• Journal of Ginseng Research
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• v.27 no.2
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• pp.78-85
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• 2003

By the Agrobacterium rhizogenes A$_4$ were induced a transformed callus of ginseng hairy root and examine to find the possibility whether it can produce certain ginsenoside. Investigations for a finding out to optimal culture medium showed that BA application is better than more factorial composition between auxins and cytokinins. For the induction of hairy root callus of ginseng, l/2 MS medium containing 1 to 3 mg of benzyladenine(BA) per liter gave the best result. The growth of ginseng hairy root callus(GHC) cultured with the 1/2MS medium supplemented with 2 mg BA/L was selected for best suspension cultures. The optimum concentration of BA for ginsenosides production was found to be 2 mg/L. Probably the inoculum size of callus plays a role with the ginsenoside production in suspension culture. AS for inoculum size of callus, 50 mg was superior to 150 mg for growth and ginsenoside production. Ginsenoside contents were highest in the suspension culture grown for four weeks under continuous light condition. In fact that continous light treatment promote strongly the synthesis of ginsenoside of the hairy root callus is first result in the world and the numerously induced root hairs of the callus leads a new method for ginsenoside production.

• 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.

• Korean Journal of Plant Tissue Culture
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• v.28 no.6
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• pp.347-351
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• 2001

Panax ginseng C.A. Meyer is a well-known Korean traditional medicine. Until now, even though major research of ginseng has been focused on the pharmacological effect, clinical application and chemical analysis of extracted secondary metabolite for several years, the physiology and gene functions of ginseng were not well known. In this research, we have developed the protein extraction methods of ginseng root and hairy root for proteome analysis in order to elucidate the gene(s) function of ginseng. Using the liquid nitrogen (equation omitted) TCA method as protein extraction method, about 660 protein spots were detected on the 2-DE gel of hairy root. Additionally, comparative analysis result of 2-DEs of ginseng root (equation omitted) hairy root suggested that proteomes of same organism could be changeable according to the culture condition, growth stages and other stimulus.