Rapid evolution of ribosomal RNA (rRNA) gene promoters often prevents their recognition in a foreign species. Unlike animal systems, we show that foreign plant rRNA gene promoters are recognized in an alien species, but tend to program transcription by a different polymerase. In plants, RNA polymerase I transcripts initiate at a TATATA element (+1 is underlined) important for promoter strength and start-site selection, However, transcripts initiate from +32 following transfection of a tomato promoter into Arabidopsis, The rRNA gene promoter of a more closely related species, Brassica oleracea, programs both +1 and +29 transcription, A paint mutation at +2 improving the identity between the Brassica and Arabidopsis promoters increases +1 transcription, indicating a role for the initiator element in species-specificity Brassica +29 transcripts can be translated to express a luciferase reporter gene, implicating RNA polymerase II, TATA mutations that disrupt TATA-binding protein (TBP) interactions inhibit +29 transcription and luciferase expression, Co-expressed TBP proteins bearing compensatory mutations restore +29 transcription and luciferase activity, suggesting a direct TBP-TATA interaction, Importantly, +1 transcription is unaffected by the TATA mutations, suggesting that in the context of pol I recognition, the TATA-containing initiator element serves a function other than TBP binding.