• Title/Summary/Keyword: hairy root callus

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Production of Saponin by Hairy Root Culture of Bupleurum falcatum L. I. Comparison of Saponin Content and Pattern in Callus, Adventitous Root, Hairy Root and Cultivated Root (시호(Bupleurum falcatum L.)의 모상근 배양을 통한 Saponin 생산 I. 캘러스, 부정근, 모상근과 재배근의 Saponin 양상과 함량 비교)

  • 안준철
    • Journal of Plant Biology
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    • v.36 no.1
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    • pp.43-49
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    • 1993
  • In order to survey possibility to produce saikosaponin from in vitro hairy root culture, culture of callus, adventitous root, and hairy root of Bupleurum falcatum L. were estabilished, and quantitative and qualitative aspects in saikosaponin extracted from these were compared with those of cultivated root. Callus grew well in MS medium containing 0.9 $\mu$M 2, 4-D. In contrast, both of adventitous root and hairy root grew well in hormone-free MS medium. However, hairy root showed more rapid growth with extensive lateral root branches, characteristics of lower content of water and softer than in adventitous root. Among the selected lines of adventitous root and hairy root were observed difference in the growth rate. Mannopine, one of opine synthesized in the transformed tissue with Agrobacterium rhizogenes. A4 were detected in the extract of hairy root lines. Pattern and content of crude saponin from adventitous and hairy root showed no difference, but somewhat difference from those of cultivated root. However, in callus, distinct production-aspect of saponin was not observed.

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Bioceramic Effects to Enhance Secondary Metabolites Production in Tissue Culture of Some Medicinal Plants

  • Kim, Yu-Jeong;Hwang, Baik;Ahn, Jun-Cheul
    • Korean Journal of Medicinal Crop Science
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    • v.12 no.2
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    • pp.118-122
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    • 2004
  • We have investigated that a couple of soft ferrite ceramic powders having a spinal structure have shown the effect on growth and secondary metabolites production of some medicinal plants cultured in vitro. The addition of the ceramic powders as bare state to culture medium has stimulated the growth of Achyranthes japonica callus and plantlet, adventitious root of Hyoscyamus niger and Platycodon grandiflorum hairy root about 65, 75, 150 and 50%, respectively. Whereas Centella asiatica callus and plantlet, Scopolia parviflora hairy root, and Hyoscyamus albus adventitious root were not affected markedly. Moreover, the ceramic powder has enhanced the growth of H. niger adventitious roots even under conditions of irradiating alone without any direct contact between ceramic powder and media. Based on growth stimulation effect, the ceramic powders have enhanced the gross production of tropane alkaloid in H. niger adventitious root, and polyacetylene in P. grandiflorum hairy root about 35 and 30%, respectively.

Production of Ginsenoside in Callus of Ginseng Hairy Roots (인삼 모상근의 캘러스를 이용한 ginsenoside 생산)

  • Kwon, Jung-Hee;Cheon, Hyun-Choon;Yang, Deok-Cho
    • 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.

Changes of Peroxidsse Isozyme Pattern and Formation of Cell Wall of Hairy Root Irlduced by Agrohacterium rhisogenes from Potato Tuber (감자(Solanum tuberosum. L) 괴경에 있어서 Agrobacterium rhizogenes에 의해 형질전환된 조직세포의 세포벽 형성과 peroxidase isozyme 양상의 변화)

  • 정현숙;김영희
    • KSBB Journal
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    • v.6 no.1
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    • pp.27-33
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    • 1991
  • Hairy root induced by A. rhizogenes from potato tuber (Solanm tuberosum L.) synthesized the agropine and mannopine which were demonstrated with paper electrophoresis. And the starch contents in hairy root were increased gradually following the developmental stage. But protein contents were decreased. The activity of ${\beta}-glucan$ synthetase II(GSII) which is related to the cell wall biosynthesis was stimulated in hairy root on the developmental stage. And chloropromazine did not influence the activity of GS II while verapamil inhibited about 60% of the activity GS II. Therefore, these results showed $Ca^{2+}$ to be effective factor in the cell wall formation. Isozyme pattern of peroxidase was investigated in the callus and hairy root induced from potato tuber.

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Physiological Studies on the Formation of Hairy Root by the Agrobacterium rhizogenes 1. Relationships between LAA content and morphological characteristics in carrot infected by A. rhizogenes (Agrobacterium rhizogenes에 의한 Hairy Root 형성에 대한 생리학적 연구)

  • Hwang, Baik;Cho, Duck-Yee;Hong, Sung-Sik
    • Journal of Plant Biology
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    • v.29 no.4
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    • pp.275-283
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    • 1986
  • Inoculation of carrot discs with Agrobacterium rhizogenes harboring Ri plasmid resulted in transformation of cells, as revealed by the tumors and hairy roots arising from them. Measurements of IAA content using HPLC indicate that it is higher in tumors and inoculated tissues than in uninoculated tissue. A lot of meristemoids and vessel elements formed I tumor tissue and the hairy roots differentiated from meristemoids. IAA content in tumor tissues is decreased with hairy root and vessel elements formation from them. Formation of wound callus in uninoculated tissues resulted on wound healing but no formation of vessel elements and hairy roots. Tumor tissues show continuous growing on hormone free medium.

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The Production of Anti-cancer Substances by in vitro Grown Cultures of Panax ginseng C.A. Meyer

  • Yang, Deok-Chun;Park, Kyung-Hwa;Kim, Yong-Hae;Yoon, Eui-Soo;Kang, Tae-Jin;Park, Kwang-Tae
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 1999.10a
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    • pp.46-57
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    • 1999
  • 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 callus and hairy roots grow vigorously and may Produce the same or more biologically active compounds for human health than natural ginseng roots. Therefore, ginseng callus and hairy roots can be used for commercial purposes. Polyacetylene, one of anti-cancer compounds in ginseng, was not detected in the callus cultured on the medium containing 2, 4-B, but cells derived from the callus growth was excellent, The ginseng calli cultured on the medium containing 2mg11 CPA and 0.05mg/1 BA was grown vigorously and produced panaxydol, one of ginseng polyacetylene. The biosynthesis of polyacetylene in callus was not affected by addition of NAA and sucrose in media. The SH medium was better than the MS medium for ginseng callus growth and biosynthesis of panaxydol. Another ginseng anti-cancer compounds, ginsenoside-Rg$_3$, Rh$_1$and Rh$_2$ were detected in ginseng hairy roots by heat treatment. Those of Panax ginseng were obtained after root disks of three-year old roots were infected with Agrobacterium rhizogenes Rl000 $A_4$T in dark condition after one month of culture. 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 5 ι Erlenmeyer flasks, 1ι roller drums, 10ι jar-fermenters, and especially in 20ι air-lift .culture vessels. All heat treatments had remarkably different ginsenoside contents. Eleven ginsenosides were determined in heat treatment, eight in freeze dried hairy roots. Contents of ginsenoside-Rbl , Rb2, Rc, Rd. Re, Rf, and Rg$_1$tested in all heat treatments were less than those of freeze dried hairy roots. Contents of glnsenoside-Rg$_2$ in heat treatment for 1 hour at 105$^{\circ}C$ was 4.92mg/g dry wt, 3.9 times higher than 1.27 mg/g dry wt of freeze dried hairy roots. The optimum condition of heat treatment for the production of ginsenoside-Rg$_3$and Rhl was 2 hours at 105$^{\circ}C$, and ginsenoside content was 2.58mg/g dry wt and 3.62mg/g dry wt, respectively. The production of ginsenoside-Rh2 was the highest in heat treatment for 2 hours at 105$^{\circ}C$ among treatments examined, and ginsenoside-Rh$_2$content was 1.08mg/g dry wt.

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Growth Characteristics of Transgenic Potato Using Wild-type Agrobacterium spp. (Agrobacterium spp. 에 의하여 형질전환된 감자조직의 생장특성)

  • 양덕춘
    • Korean Journal of Plant Resources
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    • v.11 no.2
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    • pp.176-176
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    • 1998
  • This study was carried out to obtain the information for growth characteristics of crown gall tumor and hairy root transformed by Agrobacterium spp,. on the media with phytohormones, casein hydrolysate and activated charoal. Crown gall tumors and hairly roots were formed respectively on potato tuber discs infected by tumerfaciens A ch 5 and A.rhizogenes ATCC15834. These tumors and roots could be grown on the phytohormone free media. PCR analysis of Rol C and Vir C gene fragments confirmed that crown gall root was prompted on the medium containing 2,4-D 2mg/l with casein hydrolysate lg/l. The survival ration of crown gall tumor callus derived from potato increased on medium containing the activated charcoal 0.5∼0.2mg/l because of the prevention, on the other hand, hairly roots were necrosis on the same medium. Callus derived from hairly root were excellently grown for a short time by suspension culture on liquid medium containing 2.4-d 2mg/L and casein hydrolysate lg/l.

Growth Characteristics of Transgenic Potato Using Wild-type Agrobacterium spp. (Agrobacterium spp. 에 의하여 형질전환된 감자조직의 생장특성)

  • 양덕춘
    • Korean Journal of Plant Resources
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    • v.11 no.2
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    • pp.179-187
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    • 1998
  • This study was carried out to obtain the information for growth characteristics of crown gall tumor and hairy root transformed by Agrobacterium spp,. on the media with phytohormones, casein hydrolysate and activated charoal. Crown gall tumors and hairly roots were formed respectively on potato tuber discs infected by tumerfaciens A ch 5 and A.rhizogenes ATCC15834. These tumors and roots could be grown on the phytohormone free media. PCR analysis of Rol C and Vir C gene fragments confirmed that crown gall root was prompted on the medium containing 2,4-D 2mg/l with casein hydrolysate lg/l. The survival ration of crown gall tumor callus derived from potato increased on medium containing the activated charcoal 0.5∼0.2mg/l because of the prevention, on the other hand, hairly roots were necrosis on the same medium. Callus derived from hairly root were excellently grown for a short time by suspension culture on liquid medium containing 2.4-d 2mg/L and casein hydrolysate lg/l.

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The Optimum Conditions for Induction of Ginseng Hairy Roots (인삼 모상근 유도를 위한 최적 조건)

  • 양덕춘;김용해;양덕조;신성련;최광태
    • 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.

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Transformation and Hairy Root Culture in Solanum nigrum by Agrobacterium rhizogenes (Agrobacterium rhizogenes를 이용한 까마중의 형질전환(形質轉換)과 모상근(毛狀根) 배양(培養))

  • Ko, Kyung-Soo;Heo, In-Ok;Yang, Kwan-Pal;Lee, Woon-Jin;Kim, Chang-Min;Jo, Pill-Hyeong
    • 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.

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