Phylogenetically informative length polymorphism and sequence variability in mitochondrial DNA of Australian songbirds (Pomatostomus)

Abstract

A combination of restriction analysis and direct sequencing via the polymerase chain reaction (PCR) was used to build trees relating mitochondrial DNAs (mtDNAs) from 50 individuals belonging to five species of Australian babblers (Pomatostomus). The trees served as a quantitative framework for analyzing the direction and tempo of evolution of an intraspecific length polymorphism from a third mitochondrial ancestor. The length polymorphism lies between the cytochrome b and 12S rRNA (srRNA) genes. Screening of mtDNAs within and between the five species with restriction enzymes showed that Pomatosomus temporalis was polymorphic for two smaller size classes (M and S) that are completely segregated geographically, whereas mtDNAs from the other four species were exclusively of a third, larger size (L). Inter- and intraspecific phylogenetic trees relating mtDNAs based on restriction maps, cytochrome b sequences obtained via PCR, and the two data sets combined were compared to one another statistically and were broadly similar except for the phylogenetic position of Pomatosomus halli. Both sets of phylogenies imply that only two deletion events can account for the observed intraspecific distribution of the three length types. High levels of base-substitutional divergence were detected within and between northern and southern lineages of P. temporalis, which implies a low level of gene flow between northern and southern regions as well as a low rate of length mutation. These conclusions were confirmed by applying coalescent theory to the statistical framework provided by the phylogenetic analyses.