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Electroporation‐mediated transient gene expression in isolated scutella of Hordeum vulgare
Author(s) -
Hänsch Robert,
Koprek Thomas,
Heydemann Henrike,
Mendel Ralf R.,
Schulze Jutta
Publication year - 1996
Publication title -
physiologia plantarum
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.351
H-Index - 146
eISSN - 1399-3054
pISSN - 0031-9317
DOI - 10.1111/j.1399-3054.1996.tb00671.x
Subject(s) - electroporation , hordeum vulgare , biology , gene expression , reporter gene , microbiology and biotechnology , callus , gene , protoplast , luciferase , dna , transfection , biochemistry , genetics , botany , poaceae
Electroporation was used to introduce DNA into excised scutella of immature embryos of Hordeum vulgare L. cvs Golden Promise and Delita. Using the firefly luciferase gene as reporter, parameters were analyzed for high transient gene expression while maintaining tissue viability. Enzymatic wounding was necessary for DNA uptake. The optimized protocol involves use of linearized DNA and addition of 15% (w/v) polyethylene glycol at a field strength of 950 V cm −1 and approximately 56 ms pulse length. A one‐day preculture was required for obtaining callus after electroporation. Transient gene expression was further demonstrated using the β‐glucuronidase gene. Blue spots were detected at the abaxial scutellar surface, indicating that cells competent for somatic embryogenesis are also amenable to transfection by electroporation.