Transient gene expression in electroporated intact tissues of Stylosanthes guianensis (Aubl.) Sw.

Abstract

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.