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Scientia Agricola

On-line version ISSN 1678-992X


QUECINI, Vera Maria  and  VIEIRA, Maria Lúcia Carneiro. Transient gene expression in electroporated intact tissues of Stylosanthes guianensis (Aubl.) Sw.. Sci. agric. [online]. 2001, vol.58, n.4, pp.759-765. ISSN 1678-992X.

Genetic transformation though protoplast electroporation has been established for commercially important plant species. In this work, explant sources, electric field strengths, electroporation buffers, DNA forms and osmotic pretreatment were assayed in order to optimize transient reporter gene expression in electroporated tissues of Stylosanthes guianensis, a tropical forage legume. Higher transformation rates were obtained employing cotyledonary explants and an electric field strength of 250 V cm-1. Linear plasmid DNA, chloridefree electroporation buffer and osmotic pretreatment with 1.6 mol L-1 mannitol also improved transient transformation but non-significantly. Transgene specific PCR amplification was employed to prove the transformed status of the tissues.

Keywords : GUS expression; direct DNA transfer; plant transformation; forage legume.

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