Abstract
T-DNA integration and stability were assessed in Agrobacterium-derived transgenic lettuce lines carrying a chimaeric CaMV 35S promoter-driven gus-intron gene and a chimaeric nos.nptII.nos gene. T-DNA integration was predominantly complex in transgenic plants derived from an A. tumefaciens strain carrying the supervirulent plasmid ToK47. Truncation of the right side of the T-DNA was observed in first seed generation R1 plants from one line. Complex T-DNA integration patterns did not always correlate with low transgene expression. Despite a high T-DNA copy number, ca. 30% of the lines analysed showed high transgene expression in the R1 generation. High transgene expression was stable at least to the R4 seed generation in selected high-expressing lines. Transgene expression was lost in the R2 generation in a low expressing line, while complete, heritable transgene silencing from the R0 to R2 generations was also observed in another line. A 50-fold variation in β-glucuronidase (GUS) activity and a 16-fold variation in NPTII protein content were observed between R1 plants derived from different R0 parents. Reactivation of transgene expression with 5-azacytidine in partially silenced lines indicated that low expression was associated with DNA methylation.
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McCabe, M.S., Mohapatra, U.B., Debnath, S.C. et al. Integration, expression and inheritance of two linked T-DNA marker genes in transgenic lettuce. Molecular Breeding 5, 329–344 (1999). https://doi.org/10.1023/A:1009681615365
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DOI: https://doi.org/10.1023/A:1009681615365