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Improvement of Catechin Productivity in Callus Cultures of Camellia sinensis Leaves (차나무 잎의 캘러스 배양을 통한 카테킨류의 생산성 개선)

  • Oh, Soon-Ja;Koh, Seok-Chan
    • Korean Journal of Plant Resources
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    • v.18 no.2
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    • pp.351-358
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    • 2005
  • The effects of thiamine-HCl or polyvinylpyrrolidone (PVP) on growth and catechin contents of calli from Camellia sinensis leaves were investigated to improve catechin productivity in callus cultures on the selective medium. The growth of calli was great on the proliferation medium (the MS medium with 0.2 mg/L 2,4-D and 1.0 mg/L TDZ) supplemented with $20{\sim}30\;mg/L$ thiamine-HCl. Although the caffeine content was low in calli compared to young leaves, the total content of catechins was high in calli. Particularly,(-)-epicatechin (EC) which not detected in young loaves was also detected in calli on the propagation medium containing thiamine-HCl or PVP. In addition, the (-)-epicatechingallate (ECG) content was significantly higher in calli than in young leaves. In conclusion, the proliferation medium supplemented with 30 mg/L thiamine-HCl as a vitamin source seemed to be optimal condition for the growth and catechin production in callus culture.

Electrophoresis Techniques for Identification of Callus Induced from Pinella ternata (Thunb.) Breit 1. Analysis of protein and Enzymes of Callus Induced from Pinella ternata (Thunb.) Breit (기내 배양된 반하의 전기영동에 의한 생약학적 연구 1. 전기영동법에 의한 반하 Callus의 단백질 및 효소분석)

  • Choi, Jung-Sik;Ryu, Jeom-Ho;Park, Hak-Bong;Kim, Hyung-Moo
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.33 no.2
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    • pp.122-125
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    • 1988
  • A comparative electrophoretic study on protein and several important enzymes of calli induced from stem, intercostal area and minor vein area were conducted in Pinella ternata (Thunb.) Breit. Soluble protein band patterns of calli induced from stem, intercostal area and minor vein area were distinctly different from those of the corresponding plant parts. Esterase isozyme patterns of calli induced from stem, intercostal area and minor vein area were different from those of the corresponding plant parts. Glutamate oxalo-acetate transaminase isozyme patterns of calli grown for 4 weeks induced from 3 plant parts were similar to those of the corresponding plant parts. But a high molecular weight isozyme band appeared in the calli grown for 8 weeks. Alow molecular weight isozyme band disappeared on the peroxidase isozyme patterns of calli grown for 4 weeks appeared on those of the corresponding plant parts.

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Plant regeneration from hypocotyls explants of Astragalus sinicus L. (자운영(Astragalus sinicus L.) 배축절편으로부터 식물체 재생)

  • Park, Min Sun;Choi, Pil Son
    • Journal of Plant Biotechnology
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    • v.42 no.4
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    • pp.396-400
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    • 2015
  • To investigate the optimal conditions for shoot organogenesis in Astragalus sinicus L., hypocotyl explants were cultured in Murashige & Skoog's (MS) medium supplemented with 0.1, 1.0, 2.0, or 4.0 mg/L 2,4-dichlorophenoxy acetic acid (2,4-D) for 6 weeks. 2,4-D concentration significantly effected morphogenesis: some produced calli with adventitious shoots and roots, some produced calli with adventitious roots, some produced only calli, and some produced deep-brownish calli with roots. The formation of calli with shoots and/or roots was observed at lower levels of 2,4-D, whereas calli without shoots or with deep-brownish roots were formed after treatment with higher levels of 2,4-D. Also, a shoot organogenesis ability of callus clones was observed after treatment with medium with 0.1 or 1.0 mg/L 2,4-D grown in MS medium with combinations of benzyl adenine (BA) and 2,4-D for 4 weeks. Medium with a combination of BA and 2,4-D was effective for shoot formation, whereas root organogenesis from calli decreased. The greatest amount of shoot formation was obtained when calli were cultured in MS medium containing 1.0 mg/L 2,4-D and 0.5 mg/L BA. Upon shoot transfer into 1/2 MS basal medium, plantlets developed, and the plantlets grew well in soil in a greenhouse.

Rapid Agrobacterium-mediated genetic rice transformation method using liquid media (액체배양을 이용한 단기 벼 형질전환 방법)

  • Yang, Dae-Hwa;Chang, Ahn-Cheol;Ahn, Il-Pyung;Kim, Hae-Jung;Kim, Dong-Hern;Lee, Hyo-Yeon;Suh, Seok Cheol
    • Journal of Plant Biotechnology
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    • v.40 no.1
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    • pp.37-42
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    • 2013
  • Rice is one of the most important cereal crops as a model plant for functional genomics of monocotyledons and usually transformed using Agrobacterium tumefaciens. However, the transformation's process using previous method is still time consuming and uneconomical, low efficiency. In this study, we established a new method by modifying the general Agrobacterium protocol especially in the infection and co-cultivation, Agrobacterium elimination, infected calli's selection steps using liquid media. We directly inoculated Agrobacterium containing a ZjLsL gene under the control of constitutive promoter into the 1- to 3-week-old rice calli derived from mature seeds. After 3 days of co-cultivation, the infected calli were transferred onto liquid media of Agrobacterium elimination and calli's selection for 3 days. The calli were transferred to calli's growth solid media for 14 days and then the calli transferred to shoot induction and root induction media. Putative transformants were initially selected on the medium containing phosphinothricin, and the PAT protein verified by PAT strip test. This method in this study would lead to reduction of substantial labor and time to generate transgenic plants.

Scopolamine Production in Suspension Cultures of Tumor Calli from Datura metel L. (흰독말풀(Datura metel L.)종양 캘러스의 현탁배양으로부터 Scopolamine 생성)

  • 이수경;윤길영;김용해;양덕조
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.3
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    • pp.203-211
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    • 2000
  • In this study to produce large-scale scopolamine we were examined in the tumor calli of Datura metel L. induced by Agrobacterium tumefaciens $Ery{101}$. The growth and scopolamine contents of tumor calli were higher under light condition than in dark. The optimum condition of growth and scopolamine production were fluence rate of 16 $\mu$mol $m^{-2}s^{-1}$, spectra of red light region and 16 hour light periods on 50 mL SH liquid medium in 4 weeks culture. To increase of the scopolamine contents in tumor calli, the optimum concentration of nitrogen source were 1.8 mM NH$_4$+ and 40 mM NO$_3$. The optimum elicitor concentration for production of scopolamine were 10 mg/L chitosan and 15 mg/L yeast extract. The effect of precursors were good at the concentration of 0.2 mM tropine and 0.3 mM tropic acid, respectively. In order to increase of growth and scopolamine contents. we induced mutant from Datura metel L. tumor callus. Mutants of tumor calli were obtained by 3 Krad, 4 Krad and 6 Krad of ${60}^Cor-ray$. Among them, 3 Krad tumor callus was excellent on the growth and teratoma induction. The 4 Krad tumor callus was negligible for both growth and teratoma induction. But the 6 Krad tumor callus was the best in growth and teratoma induction. The formation of the mutant calli can be enhanced through hormonal combination of 1 mg/L 2,4-dichlorophenoxyacetic acid and 0.5 mg/L benzyladenine. We carry out selection mutant tumor calli for high content tropane alkaloid and suspension cultures for scopolamine production.

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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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Light Influences, Morphogenesis and Protein Content on Callus Differentiation of Cucumber (Cucumis sativus L.) (오이 캘러스 분화시 광의 영향, 형태발생 및 단백질함량)

  • 이은모;조만현;송남현;우인식;이영복;곽상수
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.3
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    • pp.213-217
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    • 2000
  • To investigate the cucumber regeneration from embryogenic calli, shoot tips of aseptically-grown cucumber seedlings were used as explants for establishing tissue cultures. Growth and differentiation of callus were studied by using Murashige and Skoog's (MS) medium containing 0.5 to 2 mg/L 2,4-D. Plantlets were induced from shoot tip culture on the plant growth regulators-free MS medium. Non-embryogenic calli and viscous calli were induced on the medium supplemented with 0.5 to 2 mg/L 2,4-D, but embryogenic callus was not induced on the same medium. Segments (ca. 5∼10 mm) of aseptically-grown hypocotyl from five to seven days old seedlings after germination were placed on MS medium supplemented with 1 mg/L 2,4-D for 50 days. Embryogenic calli and embryoids were induced only from the seedlings grown in dark condition, and hypocotyl was placed on the media explanted in light condition. Foully-five point one percent of white fragile calli and 0.6% yellowish compact calli formed roots. Yellowish callus lines were investigated to have a considerably higher concentration of crude proteins than white callus lines. Plantlets derived from embryogenic calli or embryoids have been transferred to pots containing sterile vermiculite and perlite. Normal fruits were harvested from nutrient culture on aggregated hydroponics in the F-clean house.

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Introduction and Expression of a Thaumatin-like Protein from Rice in American Ginseng Following Agrobacterium-mediated Transformation

  • Chen, W.P.;Punja, Z.K.
    • Journal of Ginseng Research
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    • v.27 no.1
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    • pp.17-23
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    • 2003
  • Agrobacterium-mediated transformation of American ginseng (Panax quinquefolius L.) with strain LBA 4404 containing a rice thaumatin-like protein gene is described. The selectable markers used were phosphinothricin acetyltransferase and hygromycin phosphotransferase genes. Epicotyl explants from seedlings were precultured for 5-7 days on Murashige and Skoog medium with ${\alpha}$-naphthaleneacetic acid and 2,4 dichlorophenoxyacetic acid at 10 ${\mu}$M and 9 ${\mu}$M, respectively (ND medium), prior to Agrobacterium infection. The explants were immersed in a bacterial suspension for 20 min. A post-infection co-culture period of 3-4 days was provided on ND medium. Selection for transformed calli was conducted on ND medium with 20 mg/L phosphinothricin followed by 100 mg/L hygromycin over an 8-month period. it transformation frequency of 24.8% was achieved at the callusing phase. The presence of the transgenes in calli was confirmed by Southern hybridization and polymerase chain reaction analysis. The expression of the thaumatin-like protein gene in ginseng calli was demonstrated by Western blot analysis. Somatic embryos were produced from both transgenic calli and suspension cultures, and plantlets were recovered that expressed the transgenic thaumatin-like protein gene.

Design of Bi-directional Recommend Calligraphy Contents Open-market Platform (양방향 추천 캘리콘텐츠 오픈마켓 플랫폼 설계)

  • So, Kyoungyoung;Lee, Yoonhan;Moon, Kyounghee;Ko, Kwangman
    • Journal of Korea Multimedia Society
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    • v.18 no.12
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    • pp.1586-1593
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    • 2015
  • Calligraphy contents(shortly called, CalliContents) depict the feature of communication media with artistic sentences or drawings before being processed into digital contents to become printed advertisement, visual design and entertainment products. As a fast growing business model, they can be applied to every single scope of all fields these days and each application case presented excellent effects to grab consumers' attention immediately. In this paper, we designed and produced an emotional bi-directional recommendation DIY calligraphy contents platform to consume created cultural contents and boost personalized contents industry that meets consumer's needs through both wired and wireless-based software with convergence of artistic and emotional calligraphy contents and ICT. For this works, we established for DIY calligraphy consumers a foundation of a virtuous circle of the CalliContents where various CalliContents are provided in on and offline environment and a third party target is opened at the CalliContents platform