Abstract

The transport of DNA into plant mitochondria is a critical, yet incompletely understood, process essential for mitochondrial genome maintenance and function. This study investigates the role of tonoplast-derived lipid microdomains, or 'rafts,' in regulating the import of DNA into mitochondria isolated from **potato tubers** (*Solanum tuberosum*). Using an *in organello* assay, we demonstrate that the addition of tonoplast rafts significantly stimulates the activity of DNA import into the mitochondria, increasing the rate by up to eight-fold compared to control fractions. This stimulatory effect is specific to the raft fraction, suggesting that these specialized membrane microdomains, which are rich in specific lipids and proteins, carry factors that actively promote the mitochondrial DNA uptake mechanism. The findings highlight a novel regulatory link between the vacuolar membrane (tonoplast) and mitochondrial function, proposing that tonoplast rafts may serve as a source or carrier of essential components—potentially specific lipids or membrane-associated proteins—that are required for the efficient and active transport of DNA across the mitochondrial membranes. This discovery provides new insights into the complex inter-organelle communication pathways governing organelle biogenesis and genome stability in plant cells.