Phylogeny of the European species of the genus Pellia (Hepaticae; Metzgeriales) based on the molecular data from nuclear tRNA(CAA)(Leu) intergenic sequences

Species of the genus Pellia are similar to such an extent that their proper recognition based on morphological and anatomical features is difficult. T o solve this problem isozyme methods were developed. As a result of these s tudies new cryptic species were recognized and new hypotheses concerning ph ylogenetic relationship in Pellia were formed. To examine hypotheses derive d from isozyme data we decided to study species of the genus Pellia at the DNA level. Total DNA from all Polish Pellia species was isolated. The taxon omic and phylogenetic relationships in Pellia were examined using intergeni c spacer sequences between nuclear rRNA(Leu) genes organised in tandem arra ys. PCR (polymerase chain reaction) amplification of tRNA(Leu) gene spacers produced fragments of different sizes in all species and their restriction analysis showed species-specific patterns. PCR products were cloned and se quenced. Nucleotide sequence data were used for phylogenetic reconstruction . Sequence analysis confirmed previous results based on isozyme studies tha t P. endiviifolia- species A and species B as well as P. epiphylla-species S and species N (having different isozyme multilocus genotypes) are separat e sibling species. Our results also confirmed the allopolyploid character o f the only polyploid species in Pellia, P. borealis which was formed by hyb ridization of P. epiphylla- species N and P. epiphylla- species S cryptic s pecies. (C) 2001 Elsevier Science B.V. All rights reserved.