Chromosome and breeding system evolution of the genus <i>Mercurialis</i> (Euphorbiaceae): implications of ITS molecular phylogeny

The internal transcribed spacers (ITS1 and ITS2) of nuclear ribosomal DNA were amplified and sequenced from 19 samples representing all species of the genus <i>Mercurialis</i> and two outgroup species, <i>Ricinus communis</i> and <i>Acalypha hispida</i>. The length of ITS1 in the ingroups ranged from 223 to 246 bp and ITS2 from 210 to 218 bp. Sequence divergence between pairs of species ranged from 1.15% to 25.88% among the ingroup species in the combined data of ITS1 and ITS2. Heuristic phylogenetic analyses using Fitch parsimony on the combined data of ITS1 and ITS2 with gaps treated as missing generated 45 equally parsimonious trees. The strict consensus tree was principally concordant with morphological classification. Within the genus, the ITS sequences recognised two main infrageneric clades: the <i>M. perennis</i> complex including three Eurasian stoloniferous species (<i>M.␣leiocarpa, M. ovata and M. perennis</i>) and the western Mediterranean group including eight both annual and perennial species. Of the western Mediterranean clade, the annual and perennial species grouped respectively into two different groups, and the annual life form is revealed as a synapomorphic character derived from perennial, whereas in the Eurasian clade ITS phylogeny suggested <i>M. leiocarpa</i> as basal clade sister to <i>M.␣perennis</i> and <i>M. ovata</i>. ITS phylogeny failed to resolve the relationships among the different cytotypes of <i>M. ovata</i> and <i>M. perennis</i>. ITS phylogeny also suggested rapid karyotypic evolution for the genus. The karyotypic divergence among the perennial species of western Mediterranean region did not corroborate the nucleotide sequence divergence among the species. Optimisation of chromosome numbers onto the ITS phylogeny suggested x=8 to be the ancestral basic chromosome number of the genus. ITS phylogeny confirmed that the androdioecy of <i>M. ambigua</i> is derived from dioecy. The nucleotide heterozygosity and additivity in ITS sequences clearly confirm the interspecific hybridisation in the genus <i>Mercurialis</i>.