• Title/Summary/Keyword: Hairy root

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Mass Production of Gain-of-Function Mutants of Hair Roots in Ginseng (기능획득 돌연변이 인삼 모상근의 대량생산)

  • Ko, Suk-Min;In, Dong-Soo;Chung, Hwa-Jee;Choi, Dong-Woog;Liu, Jang-Ryol
    • 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.

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.

Effect of elicited by methyl jasmonate on the saponin contents of Codonopsis lanceolata (Elicitor처리가 더덕사포닌 함량에 미치는 영향)

  • Kim, Ji-Ah;Bae, Kee-Hwa;Choi, Yong-Eui
    • Journal of Plant Biotechnology
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    • v.42 no.3
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    • pp.265-270
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    • 2015
  • The roots of Codonopsis lanceolata (Campanulaceae) contain several kinds of triterpenoid saponin with high medicinal values, which have been used in traditional medicines. This study investigates the impacts of methyl jasmonate (MeJA) - adding time on the saponin synthesis and the hairy root growth of C. lanceolata. A significant decrease in major saponin (lancemaside of three kinds) content of hairy roots was observed with MeJA treatments. Contents of lancemaside A, B and E decreased about 15% more than non-treated hairy roots. In contrast, minor saponin (foetidissimoside A and aster saponin Hb) accumulation was about 15% higher than the non-treated hairy roots. These results suggest that MeJA treatment could be used in the production of teriterpene saponins.

High-Yield Production of Astragalosides from Transgenic Hairy Root Cultures of Astragalus membranaceus (형질전환된 황기 모상근으로부터 Astragalosides의 생산을 위한 연구)

  • Hwang, Sung-Jin
    • KSBB Journal
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    • v.21 no.2
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    • pp.123-128
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    • 2006
  • A transgenic hairy root clone AG-04 of Astragalus membranaceus was obtained following co-cultivation of leaf explants with Agrobacterium rhizogenes ATCC15834. This clone was examined for its growth and production of cyclolanostane-type saponins, astragalosides I, II, and III, under various culture conditions. Among the five basal media tested, Shenk and Hildebrandt(SH)(18) medium was best for roots growth and astragalosides production. The maximum root biomass was obtained at inoculum size of 500 mg FRW per flask, initial sucrose concentration of 3%, and shaking speeds of 90 rpm. The astagalosides production was promoted when the hairy root clone AG-04 was cultured at shaking speeds of 120 rpm and light irradiation of 18 h. Astragaloside contents was also stimulated with high initial sucrose concentration, and the maximum astargalosides contents of 6.21 %/g DRW was obtained at initial sucrose concentration of 6%. The addition of chitosan(100 mg/L) to the culture medium was significantly increased astragalosides production. This was 2.1 times higher than that obtained in a control culture without chitosan.

Stimulation Effect of a Soft Ferrite Ceramic Powder on Growth in Plant Cell and Tissue Cultures (연자성 세라믹 분말에 의한 식물세포 및 조직의 생장촉진 효과)

  • Ahn, Jun-Cheul;Kim, Yu-Jung;Park, Chan-Young;Hwang, Baik
    • KSBB Journal
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    • v.13 no.5
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    • pp.530-534
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    • 1998
  • The addition of the ceramic powder as state of bare in culture medium has stimulated the growth of Achyranthes japonica in both the disorganized cell and the plantlet. The grwoth rate of Hyoscyamus niger adventitious root and Pylatycodon grandiflorum hairy root was enhanced up to 100 and 250%, respectively, even though Scopolia parviflora hairy root and Hyoscyamus albus adventitious root were not. The ceramic powder has enhanced the growth of H. niger adventitious root even in a test tube immersed into its culture medium to irradiate alone without any direct contact. The ceramic powder seems to have a significant role on both the growth and the physiological action of some plants.

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Effects of Media and Elicitor on Betalain and Phytolaccosides Production in Hairy Root Cultures of Phytolacca esculenta van Houtte (자리공(Phytolacca esculenta van Houtte) 모상근의 Betalain 및 Phytolaccosides 생성에 미치는 배지 및 Elicitor 영향)

  • 양덕조;김용해;윤길영
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.6
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    • pp.507-514
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    • 1998
  • The effects of media and elicitors on betalain, phytolaccoside G and D2 production were examined in the hairy roots of Phytolacca esculenta van Houtte induced by Agrobacterium tumefaciens $A_4$T. Phytolaccoside G and D2 from Phytolacca hairy roots PEH2 were identified by TLC, HPLC, IR, Mass, $^1$H-NMR, and ^(13)C-NMR. Among the culture media tested, SH medium was the best for hairy root growth of hairy roots. White medium was the most suitable medium for betalain production, while MS medium was for phytolaccoside G and D2 production. Although the growth of hairy roots was supped by light (1,000 Lux), the production of betalain, phytolaccoside G and D2 was enhanced by the same light treatment. Addition of elicitors such as NaF, chitosan, and yeast extract to the culture medium increased the content of betalain, phytolaccoside G and D2, suggesting the importance of culture condition for the production of those componds.

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Optimum Condition for Mass Culture of Hairy Roots from Artemisia sylvatica MAX (국내 자생 그늘쑥 (Artemisia sylvatica MAX) 모상근의 대량배양을 위한 최적조건)

  • Shin, Sun-Hee;Yang, Deok-Cho
    • Journal of Plant Biotechnology
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    • v.30 no.1
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    • pp.65-71
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    • 2003
  • This research aims the production of anti-tumor substances through in vitro culture of hairy roots transformed by Agrobacterium rhizogenes in Artemisia sylvatica MAX and the effect of culture conditions on optimum growth of hairy roots. We investigated the optimum medium, pH, carbon source, sucrose, light, Fe and polyamine conditions of various lines of hairy roots (NK3, NK4, YX. NK3-10) induced from Artemisia sylvatica to increase the optimum growth of hairy roots. MS medium was the best for optimum growth of hairy root clone, NK3-S10. The optimum culture period was 4 weeks for NK3-S10. The optimum sucrose concentration was 3.5%. The optimum concentration of FeSO$_4$, spermine and spermidine was 0.1 mM, 10 mM and 100 mM, respectively.

Effects of Incorporation of Green Manure Crops on Growth and Quality in Cynanchum wilfordii Hemsley (녹비작물 토양환원이 백수오 생육 및 품질에 미치는 영향)

  • Youn, Cheol Ku;Kim, Ki Hyun;Kim, In Jae;Hong, Song Taeg;Hong, Eui Yon;Kim, Young Kuk
    • Korean Journal of Medicinal Crop Science
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    • v.25 no.2
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    • pp.115-120
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    • 2017
  • Background: The study aimed to obtain data on the effects of cultivation and soil reduction of green manure crop on the quantity and quality of organically cultivated Cynanchum wilfordii Hemsley. Methods and Results: The experiment comprised four treatments: control, hairy vetch, barley, and hairy vetch + barley (3 : 2). The plant height in the hairy vetch treatment (86.3 cm) was significantly different from that in the other treatments, whereas the stem diameter leaf area, and special product analysis division (SPAD) value did not differ across the treatments. The largest soil reduction of green manure crop was recorded in the barley treatment (440 kg/10 a), whereas the smallest was recorded in the single treatment with hairy vetch (80 kg/10 a). The hairy vetch + barley (60 : 40) treatment showed 63% more soil microorganisms than control. Radical scavenging activity estimation revealed that the total polyphenol content was highest (1,740 mg/kg), and the 2,2-diphenyl-1-picrylhydrazyl (DPPH) was 92.6% in the barley treatment. The 2,2′-azino-bis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) activation was highest in the control (51.1%), and the root yield was the highest in the barley treatment (310 kg/10 a). Conclusions: The root yield, total polyphenol content, and antioxidant activity of Cynanchum wilfordii (Maxim.) Hemsley increased in presence of the green manure crop barley.

Plant Regeneration from Hairy Root of Rehmannia glutinosa Liboschitz Transformed by Agrobacterium rhizogenes (형질전환된 지황의 모상근으로부터 식물체의 재분화)

  • Hwang, Sung-Jin
    • Korean Journal of Medicinal Crop Science
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    • v.14 no.1
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    • pp.31-35
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    • 2006
  • A protocol for plant regeneration from hairy root of Rehmannia glutinosa transformed by Agrobacterium rhizogenes ATCC15834 has been developed. Transgenic shoots were regenerated from hairy roots within 6 weeks after culture on the SH medium supplemented with 0.5 mg/l BA. Shoots were rooted on plant growth regulator free SH medium successfully. The transformed plants, which were regenerated from hairy roots, had thiner roots with extensive lateral branches, wrinkled leaves, shorter node, and grew faster compared with non-transformed plants. The biomass of the transformed plant was 1.28 g (F.W) per plant, significantly higher than the non-transformed plant (0.54 g F.W). The catalpol content in the transformed plant (0.56%) was also higher than that of the non-transformed plants (0.43%).