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KMID : 0380019950100010023
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Korean Journal of Biotechnology and Bioengineering
1995 Volume.10 No. 1 p.23 ~ p.28
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Transformation of Rice Embryogenic Cells by Electroporation Mediated Plasmid Uptake into Protoplasts
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Abstract
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Calli were induced from leaf base region of germinated rice(Oryza sativa L. cv. Nakdong) with high frequency of up to 65% on LS medium supplemented with 2.5mg/.i ;4-D in the dark at 27 C. Embryogenic calli of pale yellow, globular type were selected and used for the initiation of cell suspension cultures in AA, liquid medium with 2mg/i 2,4-D, 0.2mg/Q kinetin and 0.lmg/Q GA3. Protoplasts were isolated from the embryogenic cell suspensions after 4 months of culture and then were electroporated with 400V/cm for 1 msec. Electroporated protoplasts divided with plating efficiency of 1.1% on PCM liquid medium supplemented with 2.5mg/h´ ;4-D, 0.lmg/P kinetin and 10mM proline. The protoplasts -derived microcalli were cultured on 0.2/an membrane filter placed onto LS2.5 solid medium containing fine suspension cells as a feeder cells, for 2 weeks in the dark at 27 C. After an additional 2 weeks of culture under fluorescent light of 30±3jEE ? m-Zs-;yellow calli of 2mm diameter were transferred to regeneration medium. Shoots were produced from the green spot of protoplasts-derived calli and plants were regenerated from protoplast-derived green calli with frequencies of 11-33%.
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