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Introduction of a bean phenylalanine ammonia-lyase (PAL) transgene into tobacco plants results in epigenetic post-transcriptional gene silencing which is unstable, such that after self-pollination first generation progeny may become PAL over-expressors. The change from gene silencing to PAL over-expression is accompanied by a loss of cytosine methylation of the  PAL  transgene and reduced methylation of the endogenous tobacco  PAL2  gene, but not the  PAL1  gene. These changes are associated with the appearance of high levels of bean  PAL  and tobacco  PAL2  transcripts in the total RNA fraction from PAL over-expressing plants. However, tobacco  PAL2  transcripts are inefficiently recruited into polysomes, and tobacco PAL2 protein is not detected in leaves of PAL over-expressing or wild-type lines. Thus, in spite of the post-transcriptionally controlled increase in tobacco  PAL2  transcripts in PAL over-expressors, the increased PAL activity is primarily the result of the increase in bean  PAL  transcripts and corresponding enzymatic activity. These results reveal a complex cross-talk between expression of the  PAL  transgene and the corresponding endogenous  PAL  genes at the levels of transcription, transcript stability and polysomal recruitment during sense transgene-mediated silencing and subsequent over-expresson of PAL in tobacco.

Post-Transcriptional Regulation of Phenylalanine Ammonia-Lyase Expression in Tobacco Following Recovery from Gene Silencing