Journal of Plant Biology

, Volume 41, Issue 3, pp 193–200 | Cite as

Transformation system of rice suspension-cultured microcolonies by Electroporation

  • Joon Chul Kim
  • Seong Jin Choi
Article
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Accepted:

Abstract

For establishing a transformation system of rice (Oryza sativa), 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 μm stainless mesh. Filtered embryogenic microcolonies of 100–250 μm with pBI121 were electroporated at 400 V/cm for 1.2 ms. Four weeks after the electroporation, stable transformed calli were obtained at a frequency of 72% on the selection medium containing 100 mg/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.

Keywords

transformation system embryogenic microcolonies electroporation Oryza sativa PCR GUS expression 
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Copyright information

© The Botanical Society of Korea 1998

Authors and Affiliations

  • Joon Chul Kim
    • 1
  • Seong Jin Choi
    • 1
  1. 1.Division of Life ScienceKangwon National UniversityChunchonKorea

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