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KMID : 0368419930360030211
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Journal of Plant Biology
1993 Volume.36 No. 3 p.211 ~ p.218
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Plant Regeneration from Protoplasts Isolated through Embryogenic Cell Suspension Culture in Rice
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Jung, Byung Gyun/鄭炳均
Ahn, Jun Cheul/Ko, Kyeong Min/Kim, Young Jun/Hwang, Sung Jin/Hwang, Baik/安俊撤/高庚珉/金永俊/黃聲振/황백
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Abstract
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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℃ 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㎎/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. Multiple shoots of 4∼5 per callus emerged and were transeferred 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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