Figure 9.

Aminoacylation of trnI.2 is reduced in mterf6 plants. Analysis of trnI.2 and trnI.1 aminoacylation in soil-grown wild-type (Col-0), mterf6-1, and complemented mterf6-1 (35S:MTERF6.1 mterf6-1) plants and, furthermore, in MS medium-grown Col-0, mterf6-2, and, as a control, rnr1-3 seedlings. Northern-blot analyses were carried out with total RNA isolated with the Trizol method or isolated under conditions that preserve the aminoacylation of tRNAs. In the gel system used here (see “Materials and Methods”), the aminoacylated tRNA (loaded tRNA) migrates slower than its corresponding deacylated pre-tRNA. To visualize the difference in electrophoretic mobility, aliquots of Trizol-prepared wild-type and mterf6-1 RNA were included on the gel. Samples of 5 µg of RNA were separated by electrophoresis, blotted, and probed with [γ-³²P]ATP end-labeled oligonucleotide probes specific for trnI.2, trnI.1, and trnT.1.