Methodologies for Phylogenetic Inference


Phylogenetic inference from homologous molecular sequences is key to hypothesis testing and problem solving not only in evolutionary biology but also in a wide variety of other fields – from medicine to ecology. Model‐based phylogenetic methods rely on Markov substitution models to describe the molecular evolution as a stochastic process of character substitution over time on a phylogenetic tree relating the sequences. Model parameters are estimated with standard statistical inference methods, namely, Bayesian and maximum likelihood approaches. A typical phylogenetic analysis first infers a multiple sequence alignment. Given this alignment, a phylogenetic tree is then estimated together with branch lengths and model parameters. Ideally, the alignment and phylogeny should be estimated simultaneously, amongst others to take alignment uncertainty into account.

Key Concepts

  • Sequence Alignment estimates an assignment of homologous molecular characters, that is, nucleotides, amino acids or codons related by common ancestry.
  • Phylogenetic Tree is the hierarchical representation of evolutionary relationships between homologous molecular sequences. The leaves of the tree usually represent the present day sequences, while the internal nodes represent the common ancestors.
  • Model of Molecular Evolution is the mathematical description of the process of sequence change through time, such as character substitutions, insertions and deletions.
  • Phylogenetic Likelihood is the probability function of observing the sequence data given the model of molecular evolution and phylogenetic tree.
  • Frequentist Phylogenetic Inference relies on optimised phylogenetic likelihood to estimate parameters of a model of molecular evolution and a phylogenetic tree.
  • Bayesian Phylogenetic Inference relies on phylogenetic likelihood and a prior probability distribution of parameters to obtain a posterior probability distribution of parameters of a model of molecular evolution and a phylogenetic tree.
  • Branch Support quantifies the uncertainty of phylogenetic inference by assigning statistical confidence to the inferred partitions (i.e. clades) on a phylogenetic tree.
  • Alignment Uncertainty quantifies the statistical confidence of sequence alignment, which compounds the uncertainty of phylogenetic inference.

Keywords: sequence alignment; phylogenetic tree; substitution model; insertion–deletion model; molecular evolution; alignment uncertainty; likelihood; branch support

Figure 1.

Overview of a standard work‐flow for phylogenetic inference. Molecular sequences are modelled to be evolving on a phylogenetic tree (phylogeny) according to a character substitution and insertion–deletion (indel) process. The tree topology describes common ancestry by speciation and gene duplication. The branch lengths represent the amount of change. Characters in the observed sequences related by substitutions only are termed homologous. A multiple sequence alignment (MSA) is a matrix where each row consists of one sequence, enriched by gaps to reflect indels, so that homologous characters are matched in the same column.

Ideally, owing to their interrelation, homology and phylogeny (blue box) should be estimated jointly using a model of substitution and indel (J). Amongst other advantages, the joint approach allows taking uncertainty in the MSA into account (for instance, by marginalising it out).

However, a sequential approach (green boxes) is prevalent. Here, first an MSA is reconstructed (A), often followed by filtering (F) – that is, removing unreliable columns – promoted as a way to increase the signal‐to‐noise ratio of the MSA. The actual tree estimation step (T) typically assumes a substitution model and treats the gaps in the MSA as missing data.



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Further Reading

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Yang Z (2006) Computational Molecular Evolution. Oxford Series in Ecology and Evolution. Oxford University Press (376 pages) ISBN‐10: 0‐19‐856702‐2.

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Gil, Manuel, and Anisimova, Maria(Apr 2015) Methodologies for Phylogenetic Inference. In: eLS. John Wiley & Sons Ltd, Chichester. [doi: 10.1002/9780470015902.a0025545]