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PLANT PHYSIOLOGY , Vol 101, Issue 3 751-757, Copyright © 1993 by American Society of Plant Biologists
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MOLECULAR BIOLOGY AND GENE REGULATION |
Transformation and Regeneration of Two Cultivars of Pea (Pisum sativum L.)
H. E. Schroeder, A. H. Schotz, T. Wardley-Richardson, D. Spencer and TJV. Higgins
Division of Plant Industry, Commonwealth Scientific and Industrial Research Organization, GPO Box 1600, Canberra, ACT 2601, Australia
A reproducible transformation system was developed for pea (Pisum sativum
L.) using as explants sections from the embryonic axis of immature seeds. A
construct containing two chimeric genes, nopaline synthase-phosphinothricin
acetyl transferase (bar) and cauliflower mosaic virus 35S-neomycin
phosphotransferase (nptII), was introduced into two pea cultivars using
Agrobacterium tumefaciens-mediated transformation procedures. Regeneration
was via organogenesis, and transformed plants were selected on medium
containing 15 mg/L of phosphinothricin. Transgenic peas were raised in the
glasshouse to produce flowers and viable seeds. The bar and nptII genes
were expressed in both the primary transgenic pea plants and in the next
generation progeny, in which they showed a typical 3:1 Mendelian
inheritance pattern. Transformation of regenerated plants was confirmed by
assays for neomycin phosphotransferase and phosphinothricin acetyl
transferase activity and by northern blot analyses. Transformed plants were
resistant to the herbicide Basta when sprayed at rates used in field
practice.
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