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Regeneration of Transgenic Picea glauca, P. mariana, and P. abies after Cocultivation of Embryogenic Tissue with Agrobacterium tumefaciens
Krystyna Klimaszewska, Denis Lachance, Gervais Pelletier, Marie-Anne Lelu and Armand Séguin
In Vitro Cellular & Developmental Biology. Plant
Vol. 37, No. 6 (Nov. - Dec., 2001), pp. 748-755
Published by: Society for In Vitro Biology
Stable URL: http://www.jstor.org/stable/4293542
Page Count: 8
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Transgenic plants of three Picea species were produced after coculture of embryogenic tissue with the disarmed strain of Agrobacterium tumefaciens C58/pMP90/pBIV10 and selection on medium containing kanamycin. In addition to the nptII selectable gene (conferring resistance to kanamycin), the vector carried the uidA (β-glucuronidase) marker gene. Transformation frequencies were dependent on the species, genotype, and post-cocultivation procedure. Of the three species tested, P. mariana was transformed at the highest frequency, followed by P. glauca and P. abies. The transgenic state of the embryogenic tissue was initially confirmed by histochemical β-glucuronidase (GUS) assay followed by Southern hybridization. One to over five copies of T-DNA were detected in various transgenic lines analyzed. Transgenic plants were regenerated for all species using modified protocols for maturation and germination of somatic embryos.
In Vitro Cellular & Developmental Biology. Plant © 2001 Society for In Vitro Biology