IMPROVED ELECTROPORATION BUFFER ENHANCES TRANSIENT GENE-EXPRESSION INARACHIS-HYPOGAEA PROTOPLASTS

An electroporation medium containing 50 mM glycine or 10 mM glycylglyc ine (glygly), 70 mM potassium glutamate, and 0.4 M mannitol was evalua ted for its ability to improve transient beta-glucuronidase (GUS) expr ession in immature cotyledonary protoplasts of Arachis hypogaea L. GUS activity in electroporated protoplasts was 8- to 430-fold greater tha n that obtained using any of other four commonly employed poration med ia. Analysis of viability and histochemical staining of protoplasts in dicated that electroporation using the glycine- or glygly-based porati on medium resulted in increased protoplast viability and GUS expressio n when compared with other poration media. Replacement of glygly with MES or HEPES buffers significantly reduced the level of GUS expression in electroporated protoplasts.