Canadian Forest Service Publications

Stable genetic transformation of Picea mariana (black spruce) via particle bombardment. 1996. Charest, P.J.; Devantier, Y.; Lachance, D.A. In Vitro Cellular & Developmental Biology Plant 32(2): 91-99.

Year: 1996

Issued by: National Capital Region

Catalog ID: 10270

Language: English

Availability: Not available through the CFS (click for more information).

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Abstract

Stable genetic transformation of Picea mariana (black spruce) was obtained via particle bombardment into two target tissues, mature cotyledonary somatic embryos and suspensions from embryonal masses, with the Biolistic PDS-1000/He device. Seven transgenic embryogenic cell lines were obtained from the mature cotyledonary somatic embryos after secondary somatic embryogenesis from two different cell lines (R4F14 and 119794-014). The suspension cultures from embryonal masses produced five transgenic cell lines from one cell line (R4F14). Expression of the introduced ß-glucuronidase (GUS) and neomycin phosphotransferase II (NPT II) genes was detected by histochemistry and fluorometry, and by ELISA in 10 of the lines. Two lines showed only NPT II gene expression. Four of the five lines obtained after bombardment of suspensions of embryonal masses showed lower levels of expression of GUS and NPT II. The integration of the foreign gene was confirmed by polymerase chain reaction analyses and Southern hybridization for GUS and NPT II. The integration of the foreign genes was confirmed by polymerase chain reaction analyses and Southern hybridization for GUS and NPT II, and complex hybridization patterns were observed. The 12 transgenic lines obtained had a typical embryogenic morphology and were capable of maturation and germination. Over 40 transgenic trees were regenerated from one of the transgenic lines, and they have a normal phenotype.