Canadian Forest Service Publications
Stable genetic transformation of white pine (Pinus strobus L.) after cocultivation of embryogenic tissues with Agrobacterium tumefaciens. 1999. Levée, V.; Garin, E.; Klimaszewska, K.; Séguin, A. Molecular Breeding 5: 429-440.
Year: 1999
Issued by: Laurentian Forestry Centre
Catalog ID: 16865
Language: English
Availability: PDF (request by e-mail)
Abstract
A genetic transformation procedure for white pine has been developed after cocultivation of embryogenic tissues with Agrobacterium tumefaciens. This efficient transformation procedure led to an average of four independent transformed lines per gram of cocultivated embryogenic tissue and up to 50 transformed lines can be obtained in a routine experiment. Constructs bearing the uidA gene or the green fluorescent protein (GFP) gene were introduced and β-glucuronidase (GUS) activity was followed over time. The expression of the uidA gene was lowest with 35S-gus-intron construct and was 20-fold higher with a 35S-35S-AMV_gus::nptII_ construct. The addition of scaffold attachment region (SAR) sequences surrounding the gus:nptII fusion did not significantly enhance the GUS activity. Transformed mature somatic embryos have been germinated and plantlets are presently being acclimatized.