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KMID : 0368419980410030193
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Journal of Plant Biology
1998 Volume.41 No. 3 p.193 ~ p.200
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Transformation System of Rice Suspension-Cultured Microcolonies by Electroporation
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Kim Joon-Chul
Choi Seong-Jin
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Abstract
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For establishing a transformation system of rice (Oryza saliva), after three days of culture embryogenic suspension-cultured cell clusters were enzymatically macerated for 2 hours in electroporation buffer containing 2% cellulase and filtered through 550, 400, 250 and 100 urn stainless mesh. Filtered embryogenic microcolonies of 100-250㎛ with pBI121 were electroporated at 400 V/㎝ for 1.2㎳. Four weeks after the electroporation, stable transformed calli were obtained at a frequency of 72% on the selection medium containing 100㎎/L kanamycin. GUS gene in the genomic DNA among 20 out of 22 putative transformed calli lines were detected by PCR analysis. The expression of GUS gene into the kanamycin-resistance calli was confirmed by spectrophotometric assay and histochemical assay of GUS activity. In a histochemical study of the transgenic rice regenerants, it was shown that the GUS activity directed by the CaMV 35S promoter was localized mainly in leaf vein and root apex.
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