• Title/Summary/Keyword: Plant culture medium

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Plant Regeneration of B.juncea Through Plant Tissue and Protoplast Culture

  • Lian, Yu-Ji;Lim, Hak-Tae
    • Journal of Plant Biotechnology
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    • v.3 no.1
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    • pp.27-31
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    • 2001
  • New types of cytoplasmic male sterility in Brassica species would be very useful for the production of F$_1$, hybrid seeds. Leaves and stems of rapid cycling stock of B.juncea (CrGC4-3) containing Anand CMS were used as experimental materials for plant regeneration from protoplast culture. Very high plant regeneration rate (85%) was found in the Kao & Michayluk medium supplemented with 2 mg/L zeatin, 0.5 mg/L BAP, and 1 mg/L NAA when only leaf, not stem, segments were cultured. Protoplasts were isolated from leaves using mixtures of enzymes (1% Cellulycin, 0.5% Macerozyme) in 0.4 M mannitol and 50 mM $CaCl_2$.$2H_2$O. Mcrocalli induced from protoplasts were transferred to the shoot regeneration medium containing 2 mg/L BAP, 2 mg/L zeatin, and 0.5 mg/L NAA. After 60 days of initial protoplast culture, regenerated plantlets were obtained, acclimatized, transplanted into the pots, and grown up to the flowering stage.

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Expression of Chitinase Gene in Solanum tuberosum L.

  • Park, Kyung-Hwa;Yang, Deok-Chun;Jeon, Jae-Heung;Kim, Hyun-Soon;Joung, Young-Hee;Hyouk Joung
    • Journal of Plant Biotechnology
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    • v.1 no.2
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    • pp.85-90
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    • 1999
  • In order to protect fungal diseases, leaf disc explants of Solanum tuberosum cultivar, Belchip, was infected with an Agrobacterium MP90 strain containing chimeric gene construct, consisting of antibiotic resistance and chitinase gene driven by the CaMV 35S promoter, for transformation. Regenerated multiple shoots were selected on a medium containing kanamycin and carbenicillin after exposure to Agrobacterium. The presence and integration of the npt II and chitinase gene were confirmed by polymerase chain reaction(PCR). Northern blot analysis indicated that the genes coding for the enzyme could be expressed in potato plants. The chitinase activity of transgenic potato plants was higher than the control potato.

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The Effect of Cultural Factors on Anther Culture in Spring Wheat (Triticum aestivum)

  • Kang, Tae-Jin;Yang, Deok-Chun
    • Plant Resources
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    • v.6 no.2
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    • pp.153-158
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    • 2003
  • Effects of shaking, medium consistency and anther density on polyhaploid production in two wheat cultivars, Pavon and Chris, were studied using a modified 85D12 medium. Pavon produced more calli in shaking and more albino plants tban Chris. However, Chris produced threefold more green plants than Pavon in non-shaking treatment. More calli and green plants were derived from non-shaking treatment than those from shaking treatment. Anthers were cultured on both liquid and semi-solid 85D12 media, using two anther densities, 48 and 96 anthers per plate. Although Pavon generally produced more calli and albino plants than Chris, Chris produced more green plants than Pavon. More green plants were derived from semi-solid medium than those from liquid medium. A factor that may affect plant regeneration from anthers is the length of time on initiation medium. Most of the calli for both genotypes were transferred during the first two time periods. Fertility, as measured by seed set, was determined for all surviving regenerated plants. About 24% of Chris and Pavon anther-derived green plants in the experiment of medium consistency and anther density produced seed.

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Plant Regeneration from Shoot Tip-Derived Embryogenic Callus of Dianthus superbus

  • Lee, Eun-Ae;Kim, Joon-Chul;Kim, Won-Bae;Kim, Byeong-Hyeon;Kim, Jeong-Kan
    • Journal of Plant Biology
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    • v.37 no.3
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    • pp.381-385
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    • 1994
  • The highest degree of callus formation was obtained from the shoot tips of Dianthus superbus when cultured on the MS medium supplemented with 2.0 mg/L NAA and 0.5 mg/L BAP. Embryogenic calluses were obtained from the seperated friable calluses on MS medium containing 2.0 mg/L 2,4-D after 7-8 wk of culture. For plant regeneration, embryogenic calluses were selected and cultured on te proliferation medium. After 3 wk, somatic embryos appeared on MSK medium (0.5 mg/L NAA, 2.0 mg/L kinetin) and N6 medium (2.0 mg/L kinetin, 0.1 mg/LNAA, 0.1 mg/L 2,4-D and 2.0 g/L casein hydrolysate). When these somatic embryos were kept under continuous illumination, shoots were successfully regenerated on the both media. The shoots were rooted on MS medium supplemented with 2.0 mg/L NAA.

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Studies on The Anther Culture of Solanum nigrum L. II (Solanum nigrum L.의 약배양에 관한 연구 II)

  • 한창열
    • Journal of Plant Biology
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    • v.14 no.2
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    • pp.7-10
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    • 1971
  • Haploid cell obta-ined from microspores of Solanum nigrum were cultured on two kinds of medium, "Callus-inducing medium" and "Differentiation medium", in order to conduct histological studies of callus and examine differentiation of plantlets. On the callus-inducing medium the calli grew rapidly. The bulk of callus mass was light brown colored "Wet callus" covered on the surface with thin layers of rough and gleaming "White callus". The wet callus was consisted of parenchyma and meristematic tissues, while the white callus had no meristematic tissues. Large parenchyma cells, by successive divisions, became multicellular or poly nucleate cells which developed later to be meristematic tissues. The calli embedded on the differentiation medium quickly turned to dark brown color. Plantlets, however, came out later from these blackened callus mass. In the callus sectioned about ten weeks after imbedding on the differentiation medium, radially elongated tissue, concentric tissue, epidermis, tracheid-like structure, and plant jprimordia were observed.ure, and plant jprimordia were observed.

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Effects of Cytokinins on Secondary Embryogenesis and Plant Regeneration from Somatic Embryos of Aralia cordata Thunb. (땅두릅의 체세포배로부터 2차배 발생과 식물체 재생에 미치는 싸이토카이닌의 영향)

  • 이종천;소웅영
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.2
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    • pp.149-154
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    • 2000
  • Embryogenic suspension cultures were initiated using embryogenic callus from immature inflorescence explants (Aralia cordata Thunb.) cultured on solid MS medium containing 1 mg/L 2,4-D for 8 weeks and then the embryogenic callus was proliferated in liquid MS medium containing 1 mg/L 2,4-D. After sieving the suspensions (pore size 270$\mu$m), embryogenic cells were cultured in liquid MS medium with cytokinins (kinetin, BA, zeatin) for two weeks. When the embryogenic cells were transferred to liquid MS basal medium, primary somatic embryos were developed after 5 weeks of culture. Secondary embryos were developed directly from the primary torpedo and cotyledonary embryos cultured in solid MS basal medium. Frequency of secondary embryogenesis was higher on medium containing 2 mg/L kinetin than the other cytokinins. Plant regeneration was highly recorded by placing secondary cotyledonary embryos induced from primary cotyledonary embryos in MS medium containing 2 mg/L kinetin or 2 mg/L zeatin (25.4% and 28.6%, respectively). The plant regeneration from secordary embryos was prohibited by tertiary embryogenesis.

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Induction of Ginseng Hairy Roots And Their Possible Application To Large Scale Culture

  • Yang, Deok-Chun
    • Plant Resources
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    • v.6 no.1
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    • pp.1-6
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    • 2003
  • 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 hairy roots induced by introducing Ri-plasmid of Agrobacterium rhizogenes into genomic DNA of plant cells show vigorous growth, and the hairy roots produce the same or more saponins than natural ginseng roots. Therefore, hairy roots can be used for commercial purposes. The present study was carried out to induce hairy roots with both active growth and high saponin contents. Numerous hairy roots of Panax ginseng were obtained after root disks of three-year old roots were infected with Agrobacterium rhizogenes R1000 A4T in dark condition after one month of culture. About 3 hundred lines of hairy roots were selected according as morphological characters on medium with carbenicillin. After pre-selection of fifteen lines of hairy roots with active growth, KGHR-l and KGHR-8 lines were finally selected which had characters of high content of ginsenoside-Rd and ginsenoside-Re, respectively. 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 5L Erlenmeyer flasks, lL roller drums, 10L jar-fermenters, and especially in 20L air-lift culture vessels.

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Plant Regeneration from Embryogenic Suspension Culture of Orchardgrass (Dactylis glomerata L.) (오차드그래스의 현탁배양으로부터 부정배 형성과 식물체 재분화)

  • 이효신;권용삼;이병현;원성혜;김기용;조진기
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.20 no.1
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    • pp.7-12
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    • 2000
  • This study was carried out to improve the ability of embryo formation and the efficiency of plant regeneration from suspension cultured cells of seed derived calli of orchardgrass (Dactylis glomerata L.). The frequency of formation of round cell and cell colonies was highest at 50 days after suspension culture in $N_6$ medium supplemented with $4\;g/{\ell}$ casein hydrolysate (CH), $20\;g/{\ell}$ sucrose and $30\;g/{\ell}$ sorbitol. The highest frequency of plant regeneration and somatic embryo formation was obtained from suspension cultured cells of 60 days. Addition of CH ($4\;g/{\ell}$) in suspension culture medium gave the highest frequency of embryo formation (39.6%) and plant regeneration (73.0%).

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Effects of Phosphate, Precursor and Exogenous Berberine on the Production of Alkaloid in Plant Cell Cultures

  • Kim, Dong-Il
    • Journal of Microbiology and Biotechnology
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    • v.1 no.1
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    • pp.79-83
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    • 1991
  • The effects of phosphate concentration in the medium, feeding of biosynthetic precursor, and the addition of exogenous berberine on cell growth and berberine production were studied in cell suspension cultures of Thalictrum rugosum. The depletion of phosphate in the medium enhanced the specific productivity up to twofold with significant release of berberine into the medium. Extracellular berberine was 19% of the total in the culture without phosphate while it was 2-5% of total berberine in the culture with even low amounts of phosphate. Precursor feeding was not effective in enhancing alkaloid formation. Initial presence of exogenous berberine did not have much effect on cell growth and alkaloid production. It was found that the cells have the capacity to take up large quantities of berberine. When $500{\;}mg{\cdot}l^{-1}$ of berberine was added exogenously at the beginning, 81% of total berberine was found in the cells.

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