Short High-Voltage Pulses Promote Adventitious Shoot Differentiation from Intact Tobacco Seedlings

A modified electroporation protocol was used for studying the influence of short-duration, high-voltage pulses on the differentiation of adventitious shoots. Intact tobacco seedlings with two expanded leaves were subjected to 78-ms rectangular pulses, with field strengths varying between 50 and 400 V/cm. After electric treatment, the seedlings were explanted onto shoot differentiation medium. Inoculum final mass and regeneration capacities were significantly enhanced by electric treatment, showing a “window response” that was dependent on the field strength and had a maximum at 300 V/cm. Shoot regeneration dynamics were also accelerated in treated seedlings compared with controls. Our results revealed that short high-voltage pulses stimulate in vitro morphogenetic processes in plant systems more complex than protoplasts.