Phylogenetic relationships of colubroid snakes based on mitochondrial DNA sequences

We used a 694 bp length ofthe mitochondrial ND4 gene from 40 genera to infer phylogenetic relationships among colubroid snakes.The goals of this study were to identify conserved subsets of ND4 sequence data that could be used to address (1) which nominal higher-level colubroid taxa are monophyletic, and (2) the relationships among the monophyletic lineages identified.Use of transversions only proved the most reliable and efficient means of retrieving colubroid relationships.Transversion parsimony and neighbour-joining analyses iden* similar monophyletic higher-level taxa, but relationships among these lineages differ considerably between the two analyses.These differences were affected by the inclusion/exclusion of (1) transitions, (2) autapomorphies, and (3) the boid outgroups.Saturation effects among the transitions, uninformativeness of autapomorphies for clustering taxa, and long-branch and base-compositional problems among the boids lead us to regard the tree resulting from transversion parsimony analysis rooted with Acrochordus as the best current estimate of colubroid phylogenetic relationships.However, several aspects of this proposed phylogeny need further testing (e.g. the apparent diphyly of Natricinae is especially controversial).Relationships retrieved using all colubroid taxa are not obtained when sparsely or unevenly sampled experimental subsets of taxa are used instead, suggesting that long-branch problems can severely compromise elucidation of colubroid relationships if limited taxonomic sampling strategies are followed.We discuss the importance of this finding for previous molecular attempts to assess colubroid relationships.Our analyses confirm the historical validity of several nominal colubroid families and subfamilies, establish polyphyly of a few, but generally fail to resolve relationships among the monophyletic taxa we identify.More conservative character information will be required to confidently resolve the last issue.