Chloroplast DNA variation in a wild plant, tolmiea menziesii

Few studies of cpDNA have provided evolutionary and/or phylogenetic information at the intraspecific level. We analyzed restriction site variation using 19 endonucleases in 37 populations representing both diploid (2n = 14) and autotetraploid (2n = 28) Tolmiea menziesii. Seven restriction site mutations and five length mutations were observed. Although diploid and tetraploid Tolmiea have been intensively studied using nuclear markers, cpDNA variation provided additional evolutionary insights not revealed previously. The chloroplast genomes of diploid and tetraploid Tolmiea are as distinct as those of many pairs of congeneric species of angiosperms. Based on outgroup comparisons, the primitive chloroplast genome is present in tetraploid rather than diploid Tolmiea. These findings suggest that either: (1) diploid and tetraploid Tolmiea may have diverged since the origin of the autotetraploid, (2) the original diploid donor of the cytoplasm present in the tetraploid subsequently became extinct, or (3) the diploid was actually derived from the tetraploid via polyhaploidy. cpDNA variation also revealed that despite their close geographic proximity, diploid and tetraploid Tolmiea do not experience cytoplasmic gene flow. Last, three cytoplasmically distinct groups of diploid populations exist, two of which occupy distinct geographic areas. These findings demonstrate that, at least in some plant species, restriction fragment analysis of cpDNA can provide important evolutionary and phylogenetic information at low taxonomic levels.