The use of glufosinate as a selective agent in Agrobacterium-mediated transformation of soybean

The soybean transformation procedure using the Agrobacterium-cotyledonary n ode transformation system and the bar gene as the selectable marker coupled with glufosinate as a selective agent is described. Soybean cotyledonary e xplants were derived from 5 day old seedlings and co-cultivated with Agroba cterium tumefaciens for 3 days. Explants were cultured on Gamborg's B5 medi um supplemented with 1.67 mg l(-1) BAP and glufosinate at levels of 3.3 mg l(-1) or 5.0 mg l(-1) for 4 weeks. After 4 weeks explants were subcultured to medium containing MS major and minor salts and B-5 vitamins (MS/B5) supp lemented with 1.0 mg l(-1) zeatin-riboside, 0.5 mg l(-1) GA(3) and 0.1 mg l (-1) IAA amended with 1.7 mg l(-1) or 2.0 mg l(-1) glufosinate. Elongated s hoots were rooted on a MS/B5 rooting medium supplemented with 0.5 mg l(-)1 NAA without further glufosinate selection. Plantlets were transplanted to s oil and grown to maturity and set seed in the greenhouse. Primary transform ants and their progeny were characterized by Southern blot analysis and a l eaf paint assay.