• Title/Summary/Keyword: Plant culture medium

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Plant Regeneration from Protoplasts Isolated through Embryogenic Cell suspension Culture in Rice (벼 현탁배양을 통하여 분리된 원형질체로부터 식물체 재분화)

  • 정병균
    • Journal of Plant Biology
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    • v.36 no.3
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    • pp.211-218
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    • 1993
  • Plant regeneration was accomplished from protoplast culture of rice (Oryza sativa L. cv. Taebaeg). Embryogenic callus was induced from mature seed on MS medium containing 5 mM proline, 2.5 mg/L 2,4-D, 30 g/L sucrose in the dark at 28$^{\circ}C$ and used to establish embryogenic cell suspension culture. Suspension cells were subcultured every one week in N6 medium supplemented with 5 mM proline, 200 mg/L casein hydrolysate, 2.5 mg/L 2,4-D and amino acids of AA medium. Suspension cultures were composed of cells that were densely cytoplasmic, potentially embryogenic and were at least maintained for more than 6 months in liquid medium. Protoplasts were isolated from fast-growing suspension culture cells and cultured in a slightly modified KpR medium by mixed nurse culture. Isolated protoplasts began to divide within 5~7 days and thereafter, protoplast-derived calli were sequentially transferred to callus proliferating medium that soft agar MS medium contained 2 mg/L 2,4-D and produced distinct embryogenic cells. Microcolonies were then transferred to solid medium which consisted of MS medium containing 5 mg/L kinetin, 1 mg/L NAA, 1 mg/L ABA, 30 g/L sucrose and 10 g/L sorbitol under fluorescent light. Mulitple shoots of 4~5 per callus emerged and were transferred to hormone-free MS medium for root initiation. Thereafter, The plantlets were transferred to pots of soil to mature in the culture room.

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Production of Tropane Alkaloids by Two-stage Culture of Scopolia parviflora Nakai Adventitious Root

  • Kim, Won-Jung;Jung, Hee-Young;Min, Ji-Yun;Chung, Young-Gwan;Lee, Cheol-Ho;Choi, Myung-Suk
    • Korean Journal of Medicinal Crop Science
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    • v.12 no.5
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    • pp.372-377
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    • 2004
  • Scopolia parviflora Nakai, a rare and endangered species, is the sole plant producing tropane alkaloids (TA) among the Korean native species. In order to enhance TA productivity the SP72 root line was selected by screening 100 of root line, and the optimal culture media for root growth and TA production were investigated with the SP72 roots. Based on the several media, SH and 2B5 medium were determined as growth medium and White and NN medium as production medium. Among the four combinations of two-stage culture, 2BN (2B5 as growth medium plus NN as production medium) showed more enhanced root growth and TA production as compared with production media of White and NN medium and growth media of SH and 2B5 medium, respectively. However, bubble column bioreactor (BCB) cultures applying two-stage culture did not reveal the effective results despite of the each successful operation of two-stage culture in conical flasks and BCB cultures.

Effect of Cytokinins on the Proliferation of Multiple Shoots in Horsegrgm [Macrotyloma uniflorum (Lam.) 'Verdc.]

  • Mohamed, Shamsudeen Varisai;Jawahar, Manikam;Thiruvengadam, Muthu;Jeyakumar, Masilamani;Jayabalan, Narayanasamy Pillai
    • Journal of Plant Biotechnology
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    • v.1 no.2
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    • pp.79-83
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    • 1999
  • A method for induction of multiple shoots using cotyledonary nodes and shoot tips of Macrotyloma uniflorum (Lam.) Verdc. was described. The experiment was conducted in which shoot induction was noticed on Murashige and Skoog (MS) medium supplemented with different concentrations and combinations of four cytokinins (KIN, 2iP, Ads, BAP). These multiple shoots were later developed into normal shoots. The highest rate of shoot proliferation came from MS medium added with BAP 1.5 mg/L. The multiple shoot buds were subcultured into MS medium with BAP (0.5-1.5 mg/L) along with Ads (1.0 mg/L) and GA$_3$ (0.5 mg/L), which gave rise to the highest frequency of shoot proliferation and elongation. The shoots were rooted on MS medium supplemented with 1.75 mg/L IBA.

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Effects of Embryo Development Stage and Gelrite Concentration on Plant Regeneration in Seed Culture of Rice (벼의 종자배양에서 배의 성숙정도와 Celrie 농도가 캘러스 형성 및 식물체 재분화에 미치는 영향)

  • 권용삼;김경민;김도훈;손재근
    • Journal of Life Science
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    • v.11 no.4
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    • pp.311-315
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    • 2001
  • To identify the effects embryo developmental stage and gelrite concentration on plant regeneration in seed culture of rice, mature and immature seeds of rice were cultured on the $N_{6}$ medium supplemented with2 mg/$\ell$ 2.4-D and different levels of gelrite(0.2~1.0%). The calli formed immature embryos were produced more plants than those from mature embryos. The maximum frequency of plant regeneration was achieved in the culture of the calli of immature embryos which was harvested at the 21$^{th}$ day after pollination. The plant regeneration on the medium with gelrite was more accelerate than that on the medium with agar. The highest frequency(55%) of plant regeneration was obtained from the calli transferred to the medium with 6g/$\ell$ gelrite.

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In vitro Regeneration of Phragmites australis through Embryogenic Cultures

  • Lee Jeong-Sun;Kim Chang-Kyun;Kim In-Sung;Lee Eun-Ju;Choi Hong-Keun
    • Journal of Plant Biotechnology
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    • v.8 no.1
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    • pp.21-25
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    • 2006
  • Phragmites australis (reed) has received much attention as being one of the principle emergent aquatic plants for treating industrial and civil wastewater. Plant regeneration via plant tissue culture in p. australis was investigated. Three types of callus were identified from seeds on N6 medium plus 4.5 UM 2,4-dichlorophenoxyacetic acid (2,4-D). Yellow compact type showed the best redifferentiation, whereas white compact type and yellow friable were not competent to differentiate into plane. Solid medium culture was better than liquid suspension culture for enhancing callus growth when N6 medium supplemented with 4.5 ${\mu}M$ 2,4-D was used. Phytagel, as a gelling agent, was superior to agar in plant regeneration on N6 medium, supplemented with 9.4 ${\mu}M$ kinetin and 0.54 ${\mu}M$ $\alpha$-naphthaleneacetic acid (NAA). Transfer of the plantlets regenerated from kinetin and NAA-supplemented N6 medium to growth regulator-free MS medium enhanced the further development of the plantlets. Plantlets on subsequently grown to maturity when tansferred to potting soil. The regenerated plants exhibited morphologically normal. The system for plant regeneration of P. australis enables to propagate elite lines on a large scale for water purification in the ecosystem

Influence of pretreatment medium, fresh medium addition, and culture plate size on the production of embryos in isolated microspore culture of hot pepper (Capsicum annuum L.) (고추의 소포자 배양 시 전처리 배지, 새 배지의 첨가, 및 배양 용기의 크기가 배의 생산에 미치는 영향)

  • Park, Eun-Joon;Kim, Jin-Ae;Kim, Moon-Za
    • Journal of Plant Biotechnology
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    • v.36 no.2
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    • pp.184-192
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    • 2009
  • The influences of pretreatment medium, the addition of fresh medium, and the size of culture plate on the production of embryos were investigated in isolated microspore culture of hot pepper (Capsicum annuum L.). Among the media used for heat shock pretreatment ($32{\pm}1{^{\circ}C}$), high frequency embryo production was obtained when the sucrosestarvation medium A was used. On the other hand, neither 0.37 M mannitol solution nor NLNS medium supplemented with sucrose was not efficient for embryo production. The addition of culture medium to pretreatment media considerably decreased the embryo production even though embryo development proceeded further. The embryo production was not improved by the addition of fresh medium after 2 or 3 weeks from starting culture. Increase in the size of the culture plate from $3.5{\times}1.0$ cm to $6.0{\times}1.5$ cm improved embryo quality. These results will provide valuable information for developing an efficient microspore culture system of hot pepper for high frequency embryo production.

Callus Induction and Embryogenesis Through Pollen Culture in Paeonia albiflora PALL (작약의 화분배양에 의한 캘러스 및 배발생)

  • 김영숙;이병기
    • Korean Journal of Plant Tissue Culture
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    • v.22 no.1
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    • pp.13-17
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    • 1995
  • In order to induce haploid plant through pollen culture, pollens of Paeonia albiflora were cultured on MS liquid medium The development of micospore through pollen culture was examined The effect of low temperature (5$^{\circ}C$, 10 days) pretreatment on callus induction and embryogenesis in pollen culture was not evident Calli derived from pollen gave rise to globular embryos when transferred onto solid medium containing 0.5 mg/, 2,4-L. The effect of low temperature pretreatment and medium. combination to pollen viability was unrecognized. Pollen viability was reduced as the culture proceeded.

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Establishment of a novel plant regeneration system from suspension-derived callus in the halophytic Leymus chinensis (Trin.)

  • Sun, Yan-Lin;Hong, Soon-Kwan
    • Journal of Plant Biotechnology
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    • v.37 no.2
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    • pp.228-235
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    • 2010
  • The establishment of cell suspension culture and plant regeneration of the halophytic Leymus chinensis (Trin.) are described in this study for the first time. Callus induction solid medium containing Murashige and Shoog (MS) basic salt, $2.0\;mg\;l^{-1}$ 2,4-dichlorophenoxyacetic acid (2,4-D), and $5.0\;mg\;l^{-1}$ L-glutamic acid with $30.0\;g\;l^{-1}$ sucrose and $4.0\;g\;l^{-1}$ gelrite for solidification induced the highest rate of cell division in Type 1 callus among calli of various types. Liquid medium with the same hormone distribution was therefore, used for cell suspension culture from Type 1 callus. Over a 30 d suspension culture at 100 rpm, great amounts of biomass were accumulated, with 71.07% average daily increment and 22.32-fold total fresh weight increment. Comparison of before and after suspension culture, the distribution of different size callus pieces and the maintenance of callus type were basically unaltered, but a slight increase in relative water contents was observed. To induce the potential of plant regeneration, the directly transferring on plant regeneration solid medium containing MS basic salt, $0.2\;mg\;l^{-1}$ $\alpha$-naphthalene acetic acid (NAA), $2.0\;mg\;l^{-1}$ kinetin (Kn), and $2.0\;g\;l^{-1}$ casamino acid and indirectly transferring were simultaneously performed. Even now growth rates of suspension-derived callus on solid medium were approximately half of those of Type 1 callus, but faster somatic embryogenesis was observed. Rooting of all regenerated shoots was successfully performed on half-strength MS medium. All plants appeared phenotypically normal.

Effect of Nitrogen Source on the Cell Growth and Production of Secondary Metabolites in Bioreactor Cultures of Eleutherococcus senticosus (가시오갈피 생물반응기 배양에서 질소급원이 세포생장과 이차대사 생산에 미치는 영향)

  • Ahn, Jin-Kwon;Lee, Wi-Young;Park, So-Young
    • Journal of Plant Biotechnology
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    • v.30 no.3
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    • pp.301-305
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    • 2003
  • The effects of inorganic nitrogen sources such as KNO$_3$ and NH$_4$ NO$_3$ on cell growth and production of chlorogenic acid and eleutheroside E derivative were investigated in 5L bioreactor cultures of Eleutherococcus senticosus. The cell growth in the 1/2MS medium containing 15mMKNO$_{3}$. The fresh weight of cells harvested from bioreactor was affected by the concentration ratio of NO$_3$$^{[-10]}$ and NH$_4$$^{+}$ in culture medium. At the viewpoint of secondary metabolite production, the production of chlorogenic acid was affected by the concentration of NH$_4$$^{+}$ in the culture medium, but not by the total concentration of nitrogen sources in the culture medium. Futhermore, eleutheroside E derivative production was also affected by the concentration ratio of NO$_3$$^{[-10]}$ and NH$_4$$^{+}$ in the culture medium. Base on those results, it is suggested that cell growth and production of secondary metabolite(chlorogenic acid and eleutheroside E derivative) could be manipulated by controlling the total concentration of nitrogen sources and the concentration ratio of NO$_3$$^{[-10]}$ and NH$_4$$^{+}$ in the culture medium. medium.

Effect of Copper on Callus Formation and Plant Regeneration in Seed Culture of Rice (벼 종자배양에서 Copper가 캘러스 형성 및 식물체 재분화에 미치는 영향)

  • 권용삼;손재근
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.4
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    • pp.205-208
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    • 2001
  • This study was conducted to improve the regeneration efficiency from seed-derived calli of rice by optimizing the copper concentrations in the media. Mature seeds were cultured on MS medium supplemented with copper sulphate (0 to 5.0 mg/L) and 2 mg/L 2,4-D. Callus growth was influenced by the levels of copper sulphate containing with medium, The addition of copper sulphate (2.5 mg/L) in regeneration medium enhanced dramatically the ability of plant regeneration from seed-derived calli. The mean frequency of plant regeneration of 6 indica rices was 27.4% on medium containing copper sulphate, whereas that of the cultivars on copper-free medium was 2.4%. These results suggest that copper sulphate may have an important role in improving regeneration ability of indica rices.

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