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Abstract In the present work，floral aberrances appeared in some of CBF1 transformed tobacco plants. Comparisons of wild type plants, the main phenotype changes of transformed plants were aberrant petals, smaller flowers and shorter androeciums. Furthermore, the ratio of pollen germination was decreased and the capsule number was reduced. Using genomic DNA of each aberrant plant as a template, TAIL-PCR was performed with five arbitary degenerate (AD) primers pairing with 3 nested specific primers designed toward outside in pBI121 plasmid. After 3-step PCR reactions, the flanking sequence of each T-DNA inserting site was obtained. The PCR products were sequenced directly if it is longer than 300 bp. These sequences were searched against GenBank database using blast n or searched against pBI121 sequence using bl2seq. Analysis of flanking sequences demonstrated that they shared the same T-DNA inserting site. It was suggested that the aberrance of the transformed tobacco did not result from different sites of T-DNA insertion.
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Received: 27 March 2006
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