Stable genetic transformation ofPicea mariana (black spruce) via particle bombardment |
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Authors: | Pierre J Charest Yvonne Devantier Denis Lachance |
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Institution: | (1) Molecular Genetics and Tissue Culture Group, Petawawa National Forestry Institute, Natural Resources Canada, K0J 1J0 Chalk River, Ontario, Canada |
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Abstract: | Stable genetic transformation ofPicea 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 line were obtained from the
mature cotyledonary somatic embryos after secondary somatic embryogenesis from two different cell lines (R4F14 and 119794-014).
The suspension culture from embryonal masses produced five transgenic cell lines from one cell line (R4F14). Expression of
the introduced β-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 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. |
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Keywords: | β -glucuronidase neomycin phosphotransferase transgenic trees Biolistic |
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