Genetic transformation of Norway spruce (Picea abies (L.) Karst) using somatic embryo explants by microprojectile bombardment.
Stable transformation of Norway spruce tissue has been obtained following bombardment of mature somatic embryos with pRT99gus, a plasmid that contains neo coding for NPTII, and gusA, coding for beta-glucuronidase, both fused to the CaMV 35S promoter. At least 8 lines have been stably transformed (over 15 months in culture) following bombardment and selection on kanamycin. Polymerase chain reaction analyses showed a high frequency of cotransformation of the gusA and neo genes. The frequency of coexpression of the selected and unselected markers was 100%. DNA/DNA hybridization of one transformed line provided conclusive evidence of stable integration and showed copy numbers of over 10 plasmid sequences per genome. None of the transformed lines has remained embryogenic.
Robertson
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Dept. of Crop Science, North Carolina State University, Raleigh 27695.
Weissinger
A K
AK
Ackley
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R
Glover
S
S
Sederoff
R R
RR
D001483:Base Sequence; D004274:DNA, Recombinant / Q000235:genetics; D005821:Genetic Techniques; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D014170:Transformation, Genetic; D014197:Trees / Q000196:embryology / Q000235:genetics*
D004274:DNA, Recombinant