Retroposon insertions provide insights into deep lagomorph evolution.

The homogenous mammalian order Lagomorpha comprises about 80 species in two families, Ochotonidae (pikas) and Leporidae (rabbits and hares). However, the phylogenetic relationships among leporids are controversial. Molecular data, particularly from mitochondrial sequences, give highly homoplasious signals. To resolve the controversy between mitochondrial and nuclear data, we analyzed genomic orthologous retroposon insertion sites, a virtually homoplasy-free marker system. From a differential screen of rabbit genomic data for intronic retroposon insertions of CSINE elements, we polymerase chain reaction-amplified and sequenced 11 retroposons in eight representative lagomorphs. We found three retroposons shared among all lagomorphs but absent in outgroups, four confirmed the monophyly of leporids, and three significantly supported Pronolagus as the sister group to all other leporids. One retroposon supported the monophyly of Lepus. The position of Pronolagus outside of the remaining leporids supports the sequence-based signals of nuclear genes and clearly refutes the misleading signals of mitochondrial genes.

The complete mitochondrial sequence of Tarsius bancanus: evidence for an extensive nucleotide compositional plasticity of primate mitochondrial DNA.

Inconsistencies between phylogenetic interpretations obtained from independent sources of molecular data occasionally hamper the recovery of the true evolutionary history of certain taxa. One prominent example concerns the primate infraordinal relationships. Phylogenetic analyses based on nuclear DNA sequences traditionally represent Tarsius as a sister group to anthropoids. In contrast, mitochondrial DNA (mtDNA) data only marginally support this affiliation or even exclude Tarsius from primates. Two possible scenarios might cause this conflict: a period of adaptive molecular evolution or a shift in the nucleotide composition of higher primate mtDNAs through directional mutation pressure. To test these options, the entire mt genome of Tarsius bancanus was sequenced and compared with mtDNA of representatives of all major primate groups and mammals. Phylogenetic reconstructions at both the amino acid (AA) and DNA level of the protein-coding genes led to faulty tree topologies depending on the algorithms used for reconstruction. We propose that these artifactual affiliations rather reflect the nucleotide compositional similarity than phylogenetic relatedness and favor the directional mutation pressure hypothesis because: (1) the overall nucleotide composition changes dramatically on the lineage leading to higher primates at both silent and nonsilent sites, and (2) a highly significant correlation exists between codon usage and the nucleotide composition at the third, silent codon position. Comparisons of mt genes with mt pseudogenes that presumably transferred to the nucleus before the directional mutation pressure took place indicate that the ancestral DNA composition is retained in the relatively fossilized mtDNA-like sequences, and that the directed acceleration of the substitution rate in higher primates is restricted to mtDNA.

The colugo (Cynocephalus variegatus, Dermoptera): the primates' gliding sister?

Although a general agreement on the major groups of eutherian orders and their phylogenetic affiliations is emerging, the evolutionary affiliations among the members constituting these groups are still subject to debate. A prominent example is the recently published molecular evidence that challenges the long assumed monophyly of primates, displaying the colugo or flying lemur (Cynocephalus, Dermoptera) as a sister to anthropoid primates (Arnason et al. 2002 ) and positioning them after the prosimian primates (tarsiers and strepsirhines) split off. The phylogenetic analysis of the complete mitochondrial (mt) genome sequence of Cynocephalus variegatus presented in this study first appears to corroborate interpretations of primates as a paraphyletic group. However, more detailed analyses disclosed that mt nucleotide composition and consequently amino acid (AA) composition varied considerably among the species analyzed. This led us to assume that the flying lemur may be incorrectly grouped with anthropoids on the basis of similar mt nucleotide and AA compositions, rather than reflecting the true evolutionary relationship. To reanalyze the flying lemur's evolutionary association with other eutherian orders from a completely different molecular perspective, a molecular cladistic approach was applied. To this end, we determined the presence/absence pattern of transposable elements that provide a nearly homoplasy-free and copious source of molecular evolutionary markers, with well-defined character polarity. We could identify transposable elements, both on a multilocus and single-locus level, being present in all extant primate infraorders but absent in the flying lemur, thus clearly supporting the monophyly of primates by retropositional evidence.