Production of transgenic pea (Pisum sativum L.) plants by Agrobacterium tumefaciens - mediated gene transfer
J Puonti-Kaerlas1, T Eriksson, P Engström
1Department of Physiological Botany, University of Uppsala, Box 540, S-751 21, Uppsala, Sweden.
Abstract:
A transformation system that allows regeneration of transgenic pea plants from calli selected for antibiotic resistance was developed. Explants from axenic shoot cultures and seedling epicotyls were cocultivated with nononcogenic Agrobacterium tumefaciens strains, and transformed callus could be selected on callus-inducing media containing either 15 mg/l hygromycin or 75 mg/l kanamycin. After several passages on regeneration medium, shoot organogenesis could be reproducibly induced on hygromycin-resistant calli, but not on the calli selected for kanamycin resistance. Regenerated shoots could subsequently be rooted and transferred into the greenhouse. In addition, the effects of different callus-inducing and growth media on organogenesis were investigated. The transformation of the calli and regenerated plants was confirmed by DNA analysis.
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