• Title/Summary/Keyword: SF calli

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Plant regeneration from suspension-cultured cell clusters of Arabidopsis thaliana (애기장대(Arabidopsis thaliana)의 현탁배양세포괴로부터 식물체 재분화)

  • 김명덕;김준철;진창덕;임창진;한태진
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.3
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    • pp.195-200
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    • 1998
  • Callus induction from leaf and stem explants of Arabidopsis thaliana was successfully obtained when leaf explants were cultured on MS medium containing 2.0 mg/L 2,4-D in the dark and also, when stem explants were cultured on CP medium containing 0.5 mg/L 2,4-D and 0.1 mg/L BAP. Explant-derived sliced calli were suspension-subcultured every week in CP liquid medium with 0.5 mg/L 2,4-D and 0.1 mg/L BAP in the dark, and shoot-forming cell clusters of nodular, pale yellow and knobby type were selected after 7-8 weeks of culture. Shoots were initiated from the green spots of the selected shoot forming calli cultured on MS regeneration medium containing 0.05 mg/L IAA, 7.0 mg/L 2-iP and 30 g/L sucrose under continous illumination for four weeks. Shoot regeneration frequency (calli regenerating at least one shoot) was more than 50%. For plant regeneration, excised shoots were trnasferred to hormone free medium for root initiation after 4 weeks of culture. The regenerants were bolting after 2 weeks of culture and formed in vitro flowering buds within bracts after 4 weeks of culture.

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Plant Regeneration from Protoplasts of Suspension Cultured Cells in Arabidopsis thaliana (애기장대(Arabidopsis thaliana) 현탁배양세포의 원형질체로부터 식물체 재분화)

  • 김명덕;김준철;진창덕;임창진;한태진
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.2
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    • pp.125-131
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    • 2000
  • Protoplasts of Arabidopsis thaliana were easily isolated from the shoot-forming (SF) suspension-cultured cell clusters with 4 hours-shaking condition (40 rpm) on CPD enzyme solution containing 1% cellulase R-10, 0.25% pectolyase Y-23 and 0.5% driselase. Protoplasts were cultured on liquid KAO medium supplemented with 1 mg/L 2,4-D, 0.5 mg/L kinetin, 200 mg/L spermidine and 68 g/L glucose. Also, protoplasts were cultured on 0.2 $\mu$M membrane filter placed onto CP solid medium containing the suspension cells as feeder cells in the dark at $25^{\circ}C$ for 4 weeks. Protoplast-derived-SF calli were cultured on MS medium containing 0.05 mg/L IAA, 7 mg/L 2 ip and 30 g/L sucrose under the continuous illumination for four weeks. The frequency of shoot formation was about 60%. The regenerants were transferred into potting soil to grow mature plants. The regenerants formed the silques with seeds after 8 weeks of cultures.

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Shoot Proliferation and Plant Regeneration from Suspension-Cultured Cells of Dianthus gratianopol (패랭이꽃속 Dianthus gratianopol의 현탁배양세포로부터 Shoot 증식과 식물체 재분화)

  • Kim Joon-Chul
    • Journal of Plant Biotechnology
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    • v.32 no.4
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    • pp.301-306
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    • 2005
  • Conditions for efficient organogenesis and plant regeneration from Dianthus gratianopol suspension cultured cells were established. Shoot-forming calli of glossy surface, pale green and knobby type were selected from leaf explant-derived calli and were suspension-subcultured every week in CP liquid medium with 1.0 mg/L 2,4-D and 0.5 mg/L BAP. Combinations of 1.0 mg/L 2,4-D and 0.5 mg/L BAP, and 1.5 mg/L 2,4-D and 0.5 mg/L BAP were effective for the induction of regenerative callus from the suspension cultured cell clusters. Multiple shoot primordia were initiated from the green spots of these regenerative callus and formed shoots on MS medium with 1.0 mg/L TDZ and 0.5 mg/L PAA. Shoot regeneration frequency (calli regenerating at least one shoot) was about 87%. For plant regeneration, proliferated shoots were excised and transferred to MS medium with 0.1 mg/L NAA for root initiation after 9 weeks of culture. The regenerants were potted in soil and formed the flowering buds and petals. Also, adventitious shoots were formed from the excised green shoot primordia of regenerative callus and these shoots proliferated successfully and regenerated to whole plants.

Efficient Plant Regeneration from Mesophyll Protoplast of Arabidopsis thaliana and Morphological Characterization of Regenerants (애기장대 (Arabidopsis thaliana)의 엽육원형질체로부터 효율적인 식물체 재분화와 이들의 형태적 특성)

  • 김명덕;김준철;진창덕;임창진;한태진
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.2
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    • pp.127-132
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    • 1999
  • Protoplasts were isolated from the leaf mesophyll tissue of in vitro 4-weeks-old Arabidopsis thaliana and cultured in MS liquid medium supplemented with 2.0 mg/L NAA, 0.5 mg/L BAP and 9% mannitol in the dark at $25^{\circ}C$. When protoplast-derived microcolonies were dehydrated, the frequency of callus induction enhanced approximately 7-fold higher compared with non-dehydrated microcolonies in CP medium. Fifty callus lines were selected from dehydrated microcolonies. Shoots were efficiently initiated from the green spots of the selected shoot forming calli cultured on MS regeneration medium supplemented with 0.05 mg/L IAA, 7.0 mg/L 2-iP and 30 g/L sucrose under continous illumination for 4 weeks. Shoot regeneration frequencies (calli regenerating at least one shoot) were 3.5%~56%. Histological observations of shoot forming callus revealed that tracheary elements initiated from inner compact cells, and that meristemoids developed to shoot primordia and shoots. Roots were induced from these regenerating shoots on MS medium without phytohormones. These regenerants were successfully transplanted into potting soil. Morphological characterization of 50 protoplast-derived plants showed that the frequency of normal type was 78%.

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