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THE PLANT CELL, Vol 8, Issue 3 447-461, Copyright © 1996 by American Society of Plant Biologists
Size and Structure of Replicating Mitochondrial DNA in Cultured Tobacco Cells
D. J. Oldenburg and A. J. Bendich
Department of Botany, University of Washington, Seattle, Washington 98195-5325
The BY-2 tobacco cell line was used to study the size and structure of
replicating mitochondrial DNA (mtDNA). Approximately 70 to 90% of the newly
synthesized mtDNA did not migrate during pulsed-field gel electrophoresis.
Moving pictures of the fluorescently labeled molecules showed that most of
the immobile well-bound DNA was in structures larger than the size of the
BY-2 mitochondrial genome of ~270 kb. Most of the structures appeared as
complex forms with multiple DNA fibers. The sizes of the circular molecules
that were also observed ranged continuously from ~20 to 560 kb without
prominent size classes. Pulse-chase and mung bean nuclease experiments
showed that the well-bound DNA contained single-stranded regions and was
converted to linear molecules of between 50 and 150 kb. MtDNA replication
in plants may be initiated by recombination events that create branched
structures of multigenomic concatemers that are then processed to 50- to
150-kb subgenomic fragments.
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