Transformation and regeneration of Brassica rapa using Agrobacterium tumefaciens

• Published in: Plant cell reports Vol. 11; no. 10; pp. 499 - 505
• Main Authors: RADKE, S. E, TURNER, J. C, FACCIOTI, D
• Format: Journal Article
• Language: English
• Published: Berlin Springer 01-09-1992
• Subjects: Biological and medical sciences; Biotechnology; Fundamental and applied biological sciences. Psychology; Genetic engineering; Genetic technics; Methods. Procedures. Technologies; Transgenic animals and transgenic plants; Transgenic plants; Vegetals; Brassica rapa; Enzyme; β-D-Glucuronidase; Agrobacterium tumefaciens; Tissue culture; Kanamycin kinase; Genetic transfer; Regeneration; Plant; Hypocotyl; Cruciferae; Dicotyledones; Angiospermae; Bacteria; Spermatophyta; Transgenic plant; Rhizobiaceae; Silver Nitrates
• ISSN: 0721-7714; 1432-203X
• DOI: 10.1007/bf00236265

Transformation and regeneration procedures for obtaining transgenic Brassica rapa ssp. oleifera plants are described. Regeneration frequencies were increasedby using silver nitrate and by adjusting the duration of exposure to 2,4-D. For transformation, Agrobacterium tumefaciens strain EHA101 containing a binary plasmid with the neomycin phosphotransferase gene (NPT II) and the b-glucuronidase gene (GUS) was cocultivated with hypocotyl explants from the oilseed B. rapa cvs. Tobin and Emma. Transformed plants were obtained within three months of cocultivation. Transformation frequencies for the cultivars Tobin and Emma were 1-9%. Evidence for transformation was shown by NPT II dot blot assay, the GUS fluorometric assay, Southern analysis, and segregation of the kanamycin-resistance trait in the progeny. The transformation and regeneration procedure described here has been used routinely to transform two cultivars of B. rapa and 18 cultivars of B. napus.