Stable genetic transformation of embryogenic cultures of Abies nordmanniana (Nordmann fir or Caucasian fir) was achieved using the Biolistic® transformation technology, followed by regeneration of transgenic plants. Selection of the transgenic tissue was based on the antibiotic resistance induced by the neomycin phosphotransferase II gene (nptII), in combination with the antibiotic geneticin. Six transclones were recovered from a total of 215 bombardments. Expression of the β-glucuronidase gene (uidA) was confirmed by histochemical analysis, and expression of nptII was verified by quantification of NPTII protein by enzyme linked immunosorbent assay (ELISA). Both genes were still expressed in the embryogenic tissue after 5 yr of in vitro culture and in mature somatic embryos and plants produced from these cultures. The integration of nptII was confirmed by Southern hybridization in embryogenic tissue after 5 yr of culture. After 5 yr of growth, uidA was still expressed in needles from the transformed trees.
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1 November 2005
STABLE GENETIC TRANSFORMATION OF EMBRYOGENIC CULTURES OF ABIES NORDMANNIANA (NORDMANN FIR) AND REGENERATION OF TRANSGENIC PLANTS
JENS I. FIND,
JULIA A. CHARITY,
LYNETTE J. GRACE,
MICHEL M.M.H. KRISTENSEN,
PETER KROGSTRUP,
CHRISTIAN WALTER
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In Vitro Cellular and Developmental Biology - Plant
Vol. 41 • No. 6
November 2005
Vol. 41 • No. 6
November 2005
Abies nordmanniana
Biolistic® transformation
gene expression
genetic engineering
Nordmann fir