• Title/Summary/Keyword: ginseng root culture

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STUDIES ON THE TISSUE CULTURE OF PANAX GINSENG

  • Harn C
    • Proceedings of the Ginseng society Conference
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    • 1974.09a
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    • pp.9-22
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    • 1974
  • Unlike the tissue culture in animals and human being, in higher plants various parts of the plant are cultured for varied purposes, and they are named variously depending on which parts are used as explants or what purposes they are cultured for. Followings are some of the names of culture used frequently: organ culture, tissue culture, callus culture, single cell culture, meristem culture, mericlone culture, ovary culture, ovule culture, embryo culture, endosperm culture, anther culture, pollen culture, protoplast culture, etc.. As the names of the culture indicate, in some kinds of culture the explants used for culture are actually not tissues, but organs, single cells, or protoplasts. It seems, however, convenient to call all of the above-mentioned cultures grossly as tissue culture. Several kinds of tissue culture were attempted using Panax ginseng as material and some of the results were summarized below. 1. Callus culture After dormancy of the sed was broken, whole embryo or parts (hypocotyl, cotyledon and epicotyl) of partly grown embryo were cultured in the media supplemented with growth regulators. Rapid swelling occurred in a few weeks, but most of the swelling was observed only in the basal part of epicotyl, changes in the other parts of embryo appearing in much later stages. The swelling or increase in size, however, was resulted not from the divisions of cells, but from the mere expansion of cell. Real calli were formed about two months after inoculation of explants. Callus tissues developed from cortex, pith, and vascular bundle in the cases of hypo- and epicotyl, from mesophyl tissue in the case of cotyledon. Shoots developed more easily from cotyledons regardless of whether they are detached from or attached to the embryo proper. 2. Culture in the Knudson C medium When cotyledons, detached from or attached to the embryo proper, were cultured in the growth regulator-free Knudson C medium comprision only several kinds of mineral compounds and sucrose, shoot primordium or callus developed profusely and finally plantlets were produced directly from shoot primordium or indirectly through callus. In this medium epidermal cells as well as mesophyl cells of the cotyledon became meristematic and divided, changing into multinucleate cells or multicellular bodies, developing eventually into either shoot primordia or calli. 3. Anther culture Anthers were cultured in the media supplemented with various growth regulators applied singly or in combinations. Callus was formed mostly in the connective tissue of anther. Cells of anther wall layers changed in appearance, but no division occurred. Microspores of all stages in development were not changed, ruling out the possibility that microspore-originated callus might be formed. 4. Isolation of protoplast Protoplasts were isolated from young root, leaf, and epicotyl, using 0.7M D-mannitols as osmoticum and using macerozyme and cellulase respectively for maceration and digestion of the cell wall. Production in large number of naked intact protoplast was rather difficult as compared with other plant species. Fusion of protoplasts occurred infrequently mainly due to the fewer number of naked protoplasts in the solution.

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Effect of water potential of culture solution on water uptake, transpiration and photosynthesis of Panax ginseng (배양액(培養液)의 수분장력(水分張力)이 인삼(人蔘)의 수분흡수(水分吸收) 증산(蒸散) 및 광합성(光合成)에 미치는 영향(影響))

  • Mok, Sung-Kyun;Park, Hoon;Lee, Chong-Hwa;Son, Suk-Yeong
    • Korean Journal of Soil Science and Fertilizer
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    • v.13 no.4
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    • pp.115-118
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    • 1981
  • Effect of water potential of culture solution on photosynthesis, transpiration and water uptake was investigated using polyethylene glycol 6000. 1. Even at -0.5 bar of culture solution phothosynthesis was decreased by 20% within 1 hour. Plant in control showed 3.26% loss of initial water for 13 hours suggesting very sensitive in water uptake. 2. Relation between water potential of culture solution (${\psi}$) and water uptake amount (W) 2-year root was ${\psi}=-2.890/e^{2.796W}$ indicating that permanent wilting point will be greater than -2.89 bar. 3. Transpiration considerably decreased with the decrease of water potential and thus by 23.9% at -0.5 bar after 4 hours. 4. From the above results ginseng plant appears to have high root water potential at permanent wilting point and thus very week to water stress due to drought or high salt content in soils.

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Tissue culture of medicinal plants: micropropagation, transformation and production of useful secondary metabolites

  • Yoshimatsu, Kayo
    • 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.

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Production of taxadiene from cultured ginseng roots transformed with taxadiene synthase gene

  • Cha, Mi-Jeong;Shim, Sang-Hee;Kim, Sung-Hong;Kim, Ok-Tae;Lee, Se-Weon;Kwon, Suk-Yoon;Baek, Kwang-Hyun
    • BMB Reports
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    • v.45 no.10
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    • pp.589-594
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    • 2012
  • Paclitaxel is produced by various species of yew trees and has been extensively used to treat tumors. In our research, a taxadiene synthase (TS) gene from Taxus brevifolia was used to transform the roots of cultured ginseng (Panax ginseng C.A. Meyer) to produce taxadiene, the unique skeletal precursor to taxol. The TS gene was successfully introduced into the ginseng genome, and the de novo formation of taxadiene was identified by mass spectroscopy profiling. Without any change in phenotypes or growth difference in a TS-transgenic ginseng line, the transgenic TSS3-2 line accumulated $9.1{\mu}g$ taxadiene per gram of dry weight. In response to the treatment of methyl jasmonate for 3 or 6 days, the accumulation was 14.6 and $15.9{\mu}g$ per g of dry weight, respectively. This is the first report of the production of taxadiene by engineering ginseng roots with a taxadiene synthase gene.

Identification of Streptomyces species antagonistic to Fusarium solani or Cylindrocarpon destructans causing ginseng root rots (인삼근부(人蔘根腐) 병원균(病原菌), Fusarium solani 및 Cylindrocarpon destructans에 길항적(拮抗的)인 Streptomyces species의 분류동정(分類同定))

  • Shim, Jae-Ouk;Lee, Min-Woong
    • The Korean Journal of Mycology
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    • v.19 no.1
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    • pp.66-73
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    • 1991
  • Among 110 isolates of actinomycetes isolated from ginseng pathogen-suppressive soils, the three actinomycetes showing the effective controls to Fusarium solani or Cylindrocarpon destruc­tans causing ginseng root rots were identified according to their morphological, cultural and physio­logical characteristics on various culture media. Spore chains of K 6-2, S 2-1 and Y 2-2 were Spira (S), Retinaculum-apertum (RA) and Rectus-flexibilis (RF), respectively. Spore surfaces of K 6-2 were spiny, whereas S 2-1 and Y 2-2 were all smooth. Aerial mass colors of 3 isolates were gray series. As a result of various tests, they were identified as Streptomyces variabilis, Streptomyces virgi­niae and Streptomyces griseo/us, respectively.

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The Therapeutic Effect of Tissue Cultured Root of Wild Panax ginseng C.A. Mayer on Spermatogenetic Disorder

  • Park, Jeong-Sook;Hwang, Seock-Yeon;Lee, Won-Suk;Yu, Kee-Won;Paek, Kee-Yoeup;Hwang, Bang-Yeon;Han, Kun
    • Archives of Pharmacal Research
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    • v.29 no.9
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    • pp.800-807
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    • 2006
  • This study examined the possibility of using a tissue cultured root of wild Panax ginseng (tcwPG) as a fertility agent. The effect of tcwPG on spermatogenesis was studied using male rats. The tcwPG crude powder was administered orally to 7-week-old rats over a 6-week period. The number of sperm in the testes and epididymides was significantly higher than the control. A histological examination did not reveal any morphological changes in the testes from the tcwPG powder treated rats. Moreover, there were no significant differences in the weights of the heart, spleen, liver, kidney, brain, testes and epididymides. Oligospermia was also induced by administering 2,3,7,8-tetrachlorodaibenzo-p-dioxin (TCDD) to the rats in order to estimate the feasibility of using tcwPG as treatment for infertility caused by spermatogenic disorders. After exposing the rats to TCDD, the tcwPG saponin fraction treated rats showed some improvement in the body weight, sperm number and testis morphology. It was estimated that tcwPG had feasibility as a therapeutic agent on spermatogenic disorder.

Cell Growth Inhibitory Effect of Tissue Cultured Root of Wild Panax ginseng C.A. Mayer Extract on Various Cancer Cell Lines

  • Park, Jeong-Sook;Lee, Tae-Woong;Han, Kun
    • Natural Product Sciences
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    • v.15 no.1
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    • pp.1-7
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    • 2009
  • This study was performed to investigate the cell growth inhibitory effect of tissue cultured root of wild Panax ginseng C.A. Mayer (tcwPG). The human stomach carcinoma cell line, MKN 74, was incubated with 70% EtOH extract of tcwPG or Panax ginseng C.A. Mayer (PG) for 24 hrs. tcwPG inhibited cell growth at a concentration of $250{\mu}g/ml$. However, Panax ginseng extract did not inhibit cell growth at the same concentration. We also tested the ethyl acetate and $H_2O$ fractions of tcwPG. The inhibitory effect of the ethyl acetate fraction on cell proliferation in MKN 74 cells was more potent than that of the crude extract, and the inhibitory effect of the $H_2O$ fraction was less than that of the ethyl acetate fraction. When we separated tcwPG into polar and non-polar saponin fractions and then measured cell growth inhibition, the non-polar saponin in tcwPG exhibited cytotoxicity. To compare the effects of tcwPG on various cancer cell lines, we measured cytotoxicity in MKN 74 (stomach cancer cell line), SW 620 (colon cancer cell line) and PC 3 (prostate cancer cell line). All three cell lines showed cell growth inhibition, and the cell growth inhibitory effects were not quite different in the various cell lines. The non-polar saponins of tcwPG arrested PC 3 cells at G1-phase as did Panax ginseng.

Isolation and Phylogenetic Analysis of Acyl-CoA-binding Protein Gene from Panax ginseng C.A. Meyer (인삼으로부터 Acyl-CoA-binding Protein 유전자의 동정 및 계통적 분석)

  • 인준교;류명현;최광태;최관삼;김세영;양덕춘
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.4
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    • pp.201-204
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    • 2001
  • Acyl-CoA binding proteins (ACBP) are small highly-conserved cytosolic proteins that bind long-chain acyl-CoAs. A cDNA encoding ACBP was identified from cDNA library constructed from hairy root poly $A^{+}$ RNA in expressed sequence tags (EST) analysis. The cDNA clone was 453 bp long and carried an open reading frame of 264 bp (10 kDa). The ginseng ACBP amino acid sequence was compared with other reported plant ACBPs using the CLUSTALW. Ginseng ACBP is 89%, 81%, 80%, and 73% identical with ACBP from castor bean, lilly, Digitalis and Arabidopsis, respectively. However, ginseng ACBP is 5 amino acids smaller than Arabidopsis and rape seed ACBPs. Also there is no any known signal peptide sequence in ginseng ACBP.

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In vitro grown thickened taproots, a new type of soil transplanting source in Panax ginseng

  • Kim, Jong Youn;Kim, Dong Hwi;Kim, Young Chang;Kim, Kee Hong;Han, Jung Yeon;Choi, Yong Eui
    • Journal of Ginseng Research
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    • v.40 no.4
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    • pp.409-414
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    • 2016
  • Background: The low survival rate of in vitro regenerated Panax ginseng plantlets after transfer to soil is the main obstacle for their successful micropropagation and molecular breeding. In most cases, young plantlets converted from somatic embryos are transferred to soil. Methods: In vitro thickened taproots, which were produced after prolonged culture of ginseng plantlets, were transferred to soil. Results: Taproot thickening of plantlets occurred near hypocotyl and primary roots. Elevated concentration of sucrose in the medium stimulated the root thickening of plantlets. Senescence of shoots occurred following the prolonged culture of plantlets. Once the leaves of plantlets senesced, the buds on taproots developed a dormant tendency. Gibberellic acid treatment was required for dormancy breaking of the buds. Analysis of endogenous abscisic acid revealed that the content of abscisic acid in taproots with senescent shoots was comparatively higher than that of taproots with green shoots. Thickened taproots were transferred to soil, followed by exposure to gibberellic acid or a cold temperature of $2^{\circ}C$ for 4 mo. Cold treatment of roots at $2^{\circ}C$ for 4 mo resulted in bud sprouting in 84% of roots. Spraying of 100 mg/L gibberellic acid also induced the bud sprouting in 81% roots. Conclusion: Soil transfer of dormant taproots of P. ginseng has advantages since they do not require an acclimatization procedure, humidity control of plants, and photoautotrophic growth, and a high soil survival rate was attained.

Increasement of Secondary Metabolites and Antioxidative Activity in Panax ginseng Adventitious Root by Methyl Jasmonate (Methyl jasmonate 처리에 의한 인삼 (Panax ginseng C.A. Meyer) 부정근의 이차대사산물 및 항산화활성 증가)

  • Lim Soon;Bae Ki-Hwa;Shin Cha-Gyun;Kim Yoon-Young;Kim Yun-Soo
    • Journal of Plant Biotechnology
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    • v.32 no.3
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    • pp.225-231
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    • 2005
  • This study was initiated to investigate the impacts of methyl jasmonate (MeJA) on adventitious root growth of Panax ginseng, the production of secondary metabolites, such as ginsenosides and phenolic compounds, and antioxidative activity. Among various concentrations of MeJA, $100\;{\mu}M$ MeJA increased the ginsenosides accumulation to 26.6 mg/g dry wt, about 8 times higher than the control in ginseng adventitious roots (GAR). In addition, $50\;{\mu}M$ MeJA increased the accumulation of phenolic compounds to 0.38 mg/g dry wt, about 3 times higher than control in GAR. This MeJA treatment was more effective in conditioned medium (CM) which obtained in bioreactor after 40 days of culture than in fresh medium (FM). Treatment of $100\;{\mu}M$ MeJA in CM increased the accumulation of ginsenosides (1.7 times) and phenolic compounds (1.2 times) more than in FM, respectively. Consequently, these high accumulation of ginsenosides and phenolic compounds by MeJA led to increase the antioxidative activities expressed to the DPPH scavenging activity (over $78.3\%$). The DPPH scavenging activity in control was $45.5\%$.