• KSBB Journal
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• 제4권3호
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• pp.288-292
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• 1989

인참 근 조직에 Agrobacterium rhizogenes strain $A_4$를 접종하여 hairy roots 유도와 유도된 hairy roots의 배양 조건을 조사하였다. 48시간 배양된 균($2{\times}A92$bacteria/ml)을 접종하여 암조건($26{pm}1^{\circ}C$)하에서 배양하였을때 6-7주후 tumor가 형성되었으며, 10-12주후 hairy roots가 유도되었다. 근 년별 hairy roots 유도율은 5년근이 4, 6년근에 비하여 높았으며, 배지에 IAA, 2, 4-D, IBA 및 tryptophan 을 각각 첨가시켰을때 15-30mg/l tryptophan에서 tumor 및 hairy roots 유도율이 증가되었음을 나타내었다. 또한 유도된 hairy roots hormone-free인 RCM배지(sucrose 3%, pH 4.5)에서 배양하였다.

• 한국약용작물학회지
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• 제12권2호
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• pp.154-162
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• 2004

Hairy roots were induced from Korean ginseng (Panax ginseng C. A. Meyer) root explants and studied for their gene expression. A total of 3,000 ESTs (expressed sequence tags) from ginseng hairy root were determined and about 2,700 ESTs have a length of readable sequence, which result in 1,352 unique ESTs sequences. The 879 ESTs showed significant similarities to known nucleotide or amino acid sequences in other plant species, which were divided into eleven categories depending upon gene function. The remaining 473 sequences showed no significant matches, which are likely to be transcripts or to be matched to other organisms. The results indicated that the analysis of the ginseng hairy root ESTs by partial sequencing of random cDNA clones may be an efficient approach to isolate genes that are functional in ginseng root in a large scale. Our extensive EST analysis of genes expressed in ginseng hairy root not only contributes to the understanding of the dynamics of genome expression patterns in root organ but also adds data to the repertoire of all genomic genes.

• Biotechnology and Bioprocess Engineering:BBE
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• 제10권6호
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• pp.528-534
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• 2005

Hairy root cultures have demonstrated great promise in terms of their biosynthetic capability toward the production of secondary metabolites, but continue to constitute a major challenge with regard to large-scale cultures. In order to assess the possibility of conducting mass production of biomass, and the extraction of useful metabolites from Panax ginseng. P. ginseng hairy roots, transformed by Rhizobium rhizogenes KCTC 2744, were used in bioreactors of different types and sizes. The most effective mass production of hairy roots was achieved in several differently Sized air bubble bioreactors compared to all other bioreactor types. Hairy root growth was enhanced by aeration, and the production increased with increasing aeration rate in a 1 L bioreactor culture. It was determined that the hairy root growth rate could be substantially enhanced by increases in the aeration rate upto 0.5vvm, but at aeration rates above 0.5vvm, only slight promotions in growth rates were observed. In 20 L air bubble bioreactors, with a variety of inoculum sizes, the hairy roots exhibited the most robust growth rates with an inoculum size of 0.1% (w/v), within the range 0.1 to 0.7% (w/v). The specific growth rates of the hairy root decreased with increases in the inoculum size.

• Journal of Plant Biology
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• 제34권4호
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• pp.289-296
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• 1991

Cultures of hairy root induced from ginseng(Panax C.A. Meyer) transformed with Agrobacterium rhizogenes (strain A4, ATCC 15834) were established and morphologically two different hairy root strains (HB1, HB2) were obtained. To determine the optimum growth rate, the hairy root (HB2) was cultured in various liquid medium supplemented with or without plant growth hormone. The growth rate of hairy root cultured on MS medium was 1.3-3.1 times higher than those cultured on other media, and the optimum sucrose concentration and pH were 3-6%, 5.5-6.5, respectively. Also, the growth rate of hairy root was increased when 0.02 M ammonium nitrate, 1.2 mM potassium phosphate (monobasic) and 0.5 mg/l IBA were supplied to liquid medium. The saponin patterns and contents of hairy root (HB2) were determined by TLC and HPLC. The crude saponin contents were 4.67% and the total saponin contents were 1.0%, on dry weight basis.

• Journal of Ginseng Research
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• 제20권3호
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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 Plant Biology
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• 제36권1호
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• pp.75-81
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• 1993

Ginseng (Panax ginseng C.A. Meyer) hairy root transformed with Agrobacterium rhizogenes (strain A4) was examined cytogenetically and histologically to assess its characteristics. The optimum growth of hairy root obtained in hormone-free MS medium (sucrose 30 g/L, pH5-6) for long period cultures. All hairy root strains (HB1, HB2, HB3) had the 2n diploid number (2n=48) of chromosomes in root tip cells. There were no alteration in chromosome structure except in one cell of HB3 strain. Results of SDS-PAGE showed a few difference in pattern and number of bands between normal and hairy root of ginseng. The root anatomy of normal root and hairy root differed each other. The hairy root had a clearly defined vascular strand, and the morphology of cortical cells were disorganised with large intercellular spaces.

• Plant Resources
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• 제6권1호
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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.

• 식물조직배양학회지
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• 제28권6호
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• pp.347-351
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• 2001

인삼 (Panax ginseng C.A. Meyer)은 우리나라의 전통약재로 수 세기 동안 사용되어 왔으며, 약효 및 성분에 대한 연구가 매우 활발하게 이루어져 왔으나, 인삼의 분자생물학적, 단백질 화학적 측면에서 생리연구는 매우 미미하였다. 본 연구에서는 프로테옴믹스를 이용한 인삼 단백질 군을 연구하기 위한 첫 단계로 인삼 (천풍)및 모상근의 단백질 추출 방법을 확립하고, 이차원 전기영동을 통하여 추출방법의 유용성을 확인하였다. Homogenizer와 황산암모늄 침전을 통한 단백질 추출과 액체질소 및 TCA를 이용한 단백질 추출 방법을 수행하여 비교해 본 결과 추출 단백질 양에는 큰 차이가 없으나 이차원 전기영동 수행 시 액체질소 및 TCA를 통한 방법이 프로테옴 연구에 적합하여 훨씬 해상도가 높은 gel 이미지를 얻었을 뿐 아니라, gel당 660개 이상의 단백질 sopt을 확인하였다. 또한 인삼과 모상근의 이차원 전기영동을 비교한 결과 상당히 다른 발현 패턴을 보여, 생장조건 등 외부 환경과 생리상태의 차이에 따라, 같은 유전자를 가지고 있는 조직체라 할지라도, 매우 다른 프로테옴 (proteome)을 보일 수 있음을 보여주었다.

• Journal of Photoscience
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• 제8권1호
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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.

• KSBB Journal
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• 제5권3호
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• pp.263-268
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• 1990

인삼의 조직에 Agrobacterium rhizogenes strain 15834와 A4을 감염하여 형질전화체를 얻었다. 이는 인삼에서처럼 무균식물을 얻기어려운 경우 leaf disk 방법으로 모상근을 유도할 수 있었다. 모상근의 ginsenoside(Rg2,Rg1,Rf,Rd,Rc,Rbl, and Rb2)는 HPLC에 의해 定量하였으며, 진탕배양한 모상근의 ginsenoside의 함량은 0.34-1.19% 건량이었다. 이러한 결과는 재배 인삼과 배양 인삼의 ginsenoside의 함량에 비해 좋은 성과라고 사료된다.