Canadian Forest Service Publications

Regeneration of transgenic Picea glauca, P. mariana, and P. abies after cocultivation of embryogenic tissue with Agrobacterium tumefaciens. 2001. Klimaszewska, K.; Lachance, D.A.; Pelletier, G.; Lelu, M.-A.; Séguin, A. In Vitro Cellular and Developmental Biology Plant 37: 748-755.

Year: 2001

Issued by: Laurentian Forestry Centre

Catalog ID: 19310

Language: English

Availability: PDF (request by e-mail)

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Abstract

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 _uid_A (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.