Electroporation of isolated higher-plant mitochondria: transcripts of an introduced cox2 gene, but not an atp6 gene, are edited in organello.
To facilitate the analysis of RNA processing in plant mitochondria, a method was established for introducing foreign DNA into mitochondria isolated from maize and sorghum. This method permits the uptake of DNA of up to 11 kb into the mitochondrial matrix. In vitro incubation of maize mitochondria in a specific buffer system was found to permit splicing and editing of newly synthesized RNAs for a period of at least 7 h. This was shown both for transcripts of endogenous mitochondrial genes (atp6, cox2) and for transcripts derived from an introduced Arabidopsis thaliana cox2 gene. In contrast, when a Sorghum bicolor atp6 gene was introduced into isolated maize mitochondria, the gene was transcribed, but the RNA was not edited, although all the editing sites in maize and sorghum atp6 RNA are identical. This may indicate the presence of transcript-specific cis -acting regions in the up- or downstream untranslated sequences of the mRNA. The system described here should allow further dissection of the mechanism of RNA editing in plant mitochondria.
Staudinger
M
M
Abteilung Botanische Genetik und Molekularbiologie, Botanisches Institut und Botanischer Garten, Christian-Albrechts-Universität zu Kiel, Olshausenstrasse 40, 24098 Kiel, Germany.
Kempken
F
F
D051546:Cyclooxygenase 2; D018274:Electroporation; D018014:Gene Transfer Techniques; D007527:Isoenzymes / Q000235:genetics*; D008928:Mitochondria / Q000235:genetics*; D010940:Plant Proteins / Q000235:genetics*; D011451:Prostaglandin-Endoperoxide Synthases / Q000235:genetics*; D006180:Proton-Translocating ATPases; D012323:RNA Processing, Post-Transcriptional; D003313:Zea mays / Q000235:genetics
D007527:Isoenzymes; D010940:Plant Proteins; D051546:Cyclooxygenase 2; D011451:Prostaglandin-Endoperoxide Synthases; D006180:Proton-Translocating ATPases