Biolistic transformation of broccoli (<i>Brassica oleracea</i>var.<i>italica</i>) for transient expression of the b-glucuronidase gene

Abstract

SummaryWe report for the first time conditions for the biolistic transformation of broccoli. Efficient transient expression of the b-glucuronidase (gus) gene has been obtained following transformation of broccoli cv. Marathon F1 (Brassica oleracea var. italica) cotyledons via microparticle bombardment. The influence of several particle gun delivery parameters and pre-treatment of cotyledon leaf discs on rates of transient GUS expression have been investigated. Consistently high rates of transformation were obtained using M17 tungsten particles fired at 900 psi with a gap distance of 20 mm and a target distance of 55 mm. Bombardment of cotyledon leaf discs placed on callus induction media doubled rates of transient transformation events. Pre-culturing cotyledon leaf discs on either hormone-free media or callus induction media resulted in a decrease in transient transformation events.