RNA editing in higher plant mitochondria frequently results in the post-transcriptional conversion of specific cytidine residues to uridine residues and infrequently results in the reverse conversion. The mechanisms by which this transition could occur are deamination or transamination of the amide at C-4 of cytosine, transglycosylation of the ribosyl residue, or deletion of a CMP residue and insertion of a UMP residue. Intact maize or petunia mitochondria were supplied with [alpha]-32P-CTP to radiolabel CMP residues in the nascent transcripts, and the fate of the [alpha]-phosphate was examined by digestion of the RNA to nucleotide monophosphates and analysis by two-dimensional chromatography. A small fraction of radioactivity comigrated with UMP on two different two-dimensional thin-layer chromatography systems, and the amount of radiolabeled UMP increased between l0-min and 2-hr incubations. The conversion of cytidine-to-uridine residues was detected in the highly edited mRNA fraction but was not detected in the rRNA fraction. Recovery of radiolabeled UMP residues suggests that the [alpha]-phosphate is retained during the editing reaction. These results are consistent with either deamination or transamination, or transglycosylation mechanisms for RNA editing.