Supplementary Material for: Strong Accumulation of Chloroplast DNA in the Y Chromosomes of <b><i>Rumex acetosa</i></b> and <b><i>Silene latifolia</i></b> P.Steflova R.Hobza B.Vyskot E.Kejnovsky 2013 Chloroplast DNA (cpDNA) sequences are often found in plant nuclear genomes, but patterns of their chromosomal distribution are not fully understood. The distribution of cpDNA on the sex chromosomes can only be studied in dioecious plant species possessing heteromorphic sex chromosomes. We reconstructed the whole chloroplast genome of <i>Rumex acetosa</i> (sorrel, XY<sub>1</sub>Y<sub>2</sub> system) from next generation sequencing data. We systematically mapped the chromosomal localization of various regions of cpDNA in <i>R. acetosa</i> and in <i>Silene latifolia</i> (white campion, XY system) using fluorescence in situ hybridization. We found that cpDNA was accumulated on the Y chromosomes of both studied species. In <i>R. acetosa</i>, the entire Y chromosome gathered all parts of cpDNA equally. On the contrary, in <i>S. latifolia</i>, the majority of the cpDNA, corresponding to the single copy regions, was localized in the centromere of the Y chromosome, while the inverted repeat region was present also in other loci. We found a stronger accumulation of cpDNA on the more degenerated Y<sub>1</sub> and Y<sub>2</sub> chromosomes of <i>R. acetosa</i> than in evolutionary younger <i>S. latifolia</i> Y chromosome. Our data stressed the prominent role of the Y chromosome centromere in cpDNA accumulation.