• Title/Summary/Keyword: hairy root culture

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Carotenoid Production of Transformed Carrot Hairy Root and Its Culture in Bioreactor (형질전환된 당근 모상근의 Carotenoid 생산과 생물반응기에서의 배양)

  • 고경민
    • Journal of Plant Biology
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    • v.35 no.4
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    • pp.365-370
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    • 1992
  • 다양한 배양조건을 이용하여 당근 모상근의 생장과 carotenoid 함량을 조사하였으며, 이러한 배양조건을 airlift형생물 반응기에 적용하여 모상근의 다량 배양을 시도하였다. 고형배지 배양에서의 carotenoid 함량은 광조건에서 배양된 모상근이 암조건에서 배양된 모상근보다 10배 정도 증가하였으며, 모상근의 생장은 광조건이 암조건보다 낮은 수준이었다. 공기 유출기 배양에서는 모상근이 광조건과 암조건 모두에서 매우 빠르게 생장하여 생줄량의 차이가 없었으며, 모상근의 생장과 carotenoid 함량에 있어서 빛의 조사는 모상근의 전체적인 생중량을 증가시키기 보다는 배양 후기 생장단계에서 carotenoid 함량을 증가시켜서 암조건에 비하여 10배 정도의 carotenoid(11 $\mu\textrm{g}$/g)를 생산하였다. 모상근을 생물 반응기에서 배양하였을 때, 모상근이 지수함수적으로 빠르게 생장하여 배양 25일 동안에 처음 무게의 11배를 나타내었다. 그리고 pH는 처음 5.5에서 5일후 4.9로 떨어지다가 그 이후에 일정한 수준(pH 4.8)을 보여주었으며, 광주기와 광도을 조절하지 않은 상태에서의 carotenoid 함량은 2$\mu\textrm{g}$/g으로 나타났다.

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Analysis and Culture Conditions for Biosynthesis of Polyacetylene from Callus of Ginseng Superior lines (인삼 육성계통 캘러스부터 Polyacetylene의 분석 및 생합성에 미치는 배양조건)

  • 양덕춘;송남현;양계진;배창휴
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.3
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    • pp.123-128
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    • 2001
  • In order to develop the biotechnological methods for the mass production of anticancer compounds from tissue culture of Panax ginseng C.A. Mayer, these studies were carried out for the selection of ginseng cell lines containing higher concentration of polyacetylene compounds and optimal condition for their biosynthesis. Panaxynol, one of ginseng polyacetylene, was not detected in any callus induced from ginseng superior cell lines cultured on MS medium supplemented with $\beta$-chlorophenoxy acetic acid (CPA). Panaxydol, another one of polyacetylene and anticancer compounds, were detected in calli of 5 cell lines by thin layer chromatogram and gas chromatogram. Among the 18 ginseng superior lines, the cell line 30201 has higher content of panaxydol. Especially, panaxydol was not detected in the callus induced from cell line 10301 which cultured on the medium containing CPA only, however, it was detected on the same callus cultured on mixed medium containing CAP 2 mg/L and BA 0.05 mg/L. SH medium was better than MS medium for ginseng callus growth and biosynthesis of polyacetylene, and also found that it was not effected by NAA and sucrose concentration in the culture medium.

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Optimization of Betacyanin Production by Red Beet (Beta vulgaris L.) Hairy Root Cultures. (Red Beet의 모상근 배양을 이용한 천연색소인 Betacyanin 생산의 최적화)

  • Kim, Sun-Hee;Kim, Sung-Hoon;Lee, Jo-No;An, Sang-Wook;Kim, Kwang-Soo;Hwnag, Baik;Lee, Hyeong-Yong
    • Microbiology and Biotechnology Letters
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    • v.26 no.5
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    • pp.435-441
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    • 1998
  • Optimal conditions for the production of natural color, betacyanin were investigated by varying light intensity, C/N ratio, concentrations of phosphate and kinds of elicitors. Batch cultivation was employed to characterize cell growth and betacyanin production of 32 days. The maximum specific growth rate, ${\mu}$$\sub$max/, was 0.3 (1/day) for batch cultivation. The maximum specific production rate, q$\^$max/$\sub$p/, was enhanced 0.11 (mg/g-cell/day) at 3 klux. A light intensity of 3 klux was shown to the best for both cell growth and betacyanin production. The maximum specific production rate was 0.125 (mg/g-cell/day) at 0.242 (1/day), the maximum specific growth rate. The dependence of specific growth rate on the light lintensity is fit to the photoinhibition model. The correlation between ${\mu}$ and q$\sub$p/ showed that the product formation parameters, ${\alpha}$ and ${\beta}$$\sub$p/ were 0.3756 (mg/cell) and 0.001 (mg/g-cell/day), respectively. The betacyanin production was partially cell growth related process, which is different from the production of a typical product in plant cell cultures. In C/N ratio experiment, high carbon concentration, 42.1 (w/w) improved cell growth rate while lower concentration, 31.6 (w/w) increased the betacyanin production rate. The ${\mu}$$\sub$max/ and q$\^$max/$\sub$p/ were 0.26 (1/day) and 0.075 (mg/g-cell/day), respectively. Beta vulgaris L. cells under 1.25 mM phosphate concentration produced 10.15 mg/L betacyanin with 13.46 (g-dry wt./L) of maximum cell density. The production of betacyanin was elongated by adding 0.1 ${\mu}$M of kinetin. This also increased the cell growth. Optimum culture conditions of light intensity, C/N, phosphate concentration were obtained as 5.5 klux, 27 (w/w), 1.25 mM, respectively by the response surface methodology. The maximum cell density, X$\sub$max/, and maximum production, P$\sub$max/, in optimized conditions were 16 (g-dry wt./L), 12.5 (mg/L) which were higher than 8 (g-dry wt./L), 4.48 (mg/L) in normal conditions. The ${\mu}$$\sub$max/ and q$\^$max/$\sub$p/ were 0.376 (1/day) and 0.134 (mg/g-cell/day) at the optimal condition. The overall results may be useful in scaling up hairy root cell culture system for commercial production of betacyanin.

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Effects of Organic Fertilizers and Green Manure Crops on Growth Responses and Yields of Astragalus membranaceus Bunge (황기의 녹비작물 작부체계 활용 유기재배 시 생장반응과 수량에 관한 연구)

  • Song, Beom-Heon;Lee, Kyung-A;Chang, Yoon-Kee;Kim, Young-Gook;Ahn, Tae-Jin;Ahn, Young-Seob;Park, Chung-Beom
    • Korean Journal of Medicinal Crop Science
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    • v.19 no.2
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    • pp.83-89
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    • 2011
  • This studies were conducted to investigate the growth responses and yields of Astragalus membranaceus Bunge which were grown with organic cultivation using two organic fertilizers and two green manure crops of hairy vetch and rye in 2008 and 2009. The lengths of stems and roots were not clearly different between organic and conventional cultivations. The root diameters and the branch numbers were slightly thicker and more in all treatments of organic cultivation than that of conventional cultivation in 2008, although they were not significantly different between two cultivating methods in 2009. The dry weights of shoot and root were heavier on organic cultivation than those on conventional cultivation. The T/R ratios were commonly higher in organic culture, showing more clearly in 2008 than 2009. The yields were generally higher in organic cultivation than those in conventional cultivation in 2008 although the effect did not clear on yield in 2009. The organic cultivation using of organic fertilizers and green manure crops would be new method to have safety and qualitative products. The techniques of organic cultivation for Astragalus membranaceus Bunge would be studied more on utilizing the natural organic resources.

Isolation of Hypervirulent Agrobacterium spp from Korea and Application for Transformation of Tobacco (한국산 고감염 Agrobacterium spp의 분리 및 연초의 형질전환에 이용)

  • 양덕춘;정재훈;이정명
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.3
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    • pp.207-217
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    • 1998
  • Total of 78 strains were characterized based on the morphological characteristics of colonies isolated on Schroth, and New & Kerr's media for selection of hypervirulent wild-type Agrobacterium spp from galls, hairy root-like process and soil of Populus, Malus, Salix and Diopyros in Korea. Among them, 48 strains were able to induce tumors in carrot disc. Hypervirulent A. tumefaciens SP101 and SM042 were identified as biotype 1 and biotype 2, respectively, These strains formed fast growing, larger tumors as compared to those induced by other strains. The binary vector pGA643 with kanamycin resistant gene was mobilized from E. coli MC100 into A. tumefaciens strain SM042 isolated from soil, and/or disarmed vector PC2760 using a triparental mating method with E. coli HB101/pRK2013, and transconjugants, A. tumefaciens SM643 and PC643 were obtained in minimal media containing kanamycin and tetracycline. Tobacco tissues were cocultivated with conjugant Agrobacterium and then transferred to selective medium with 2,4-D and kanamycin to induce the transformants. Calli were formed more efficiently in cocultivation with A. tumefaciens SM643 than that with A. tumefaciens PC643. Most of calli transformed with A. tumefaciens PC643 were friable and regenerated into normal plantlets, while the calli transformed with A. tumefaciens SM643 were compact, hard, and mixed with friable calli. The friable calli formed normal shoots, while compact calli did not form shoots but only grew to typical compact tumor calli. When the shoots formed directly from tobacco stems without callus induction after transformation by A. tumefaciens SM643 with wild-type Ti-plasmid, normal transformed plants can be induced without using disarmed Ti-plasmid.

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