## Reconstructing phylogenies from molecular data

Traditionally, phylogenies have been constructed from morphological data, but following the growth of genetic information it has become common practice to construct phylogenies based on molecular data, known as*molecular phylogeny*. The data is most commonly represented in the form of DNA or protein sequences, but can also be in the form of e.g. restriction fragment length polymorphism (RFLP).

Methods for constructing molecular phylogenies can be distance based or character based.

**Distance based methods**

Two common algorithms, both based on pairwise
distances, are the UPGMA and the Neighbor Joining algorithms. Thus, the first step in these analyses is to
compute a matrix of pairwise distances between OTUs from their sequence differences. To correct for multiple substitutions it is common to use distances corrected by a model of molecular evolution such as the Jukes-Cantor model [Jukes and Cantor, 1969].

**UPGMA.** A simple but popular
clustering algorithm for distance data is Unweighted Pair Group
Method using Arithmetic averages (UPGMA) ([Michener and Sokal, 1957],
[Sneath and Sokal, 1973]). This method works by initially having all
sequences in separate clusters and continuously joining these. The
tree is constructed by considering all initial clusters as leaf
nodes in the tree, and each time two clusters are joined, a node is
added to the tree as the parent of the two chosen nodes. The
clusters to be joined are chosen as those with minimal pairwise
distance. The branch lengths are set corresponding to the distance
between clusters, which is calculated as the average distance
between pairs of sequences in each cluster.

The algorithm assumes that the distance data has
the so-called *molecular clock* property i.e. the divergence of
sequences occur at the same constant rate at all parts of the tree. This means that the leaves of
UPGMA trees all line up at the extant sequences and that a root is estimated as part of the procedure.

**Figure 21.8:** *Algorithm choices for phylogenetic inference. The bottom shows a tree found by the neighbor joining algorithm, while the top shows a tree found by the UPGMA algorithm. The latter algorithm assumes that the evolution occurs at a constant rate in different lineages.*

**Neighbor Joining.**
The neighbor joining
algorithm,[Saitou and Nei, 1987], on the other hand, builds a tree
where the evolutionary rates are free to differ in different
lineages, i.e., the tree does not have a particular root. Some
programs always draw trees with roots for practical reasons, but for
neighbor joining trees, no particular biological hypothesis is
postulated by the placement of the root. The method works very much
like UPGMA. The main difference is that instead of using pairwise
distance, this method subtracts the distance to all other nodes from
the pairwise distance. This is done to take care of situations where
the two closest nodes are not neighbors in the "real" tree. The
neighbor join algorithm is generally considered to be fairly good
and is widely used. Algorithms that improves its cubic time
performance exist. The improvement is only significant for quite
large datasets.

**Character based methods.** Whereas the distance based methods
compress all sequence information into a single number, the
character based methods attempt to infer the phylogeny based on all
the individual characters (nucleotides or amino acids).

**Parsimony.** In parsimony based methods a number of sites are
defined which are informative about the topology of the tree. Based
on these, the best topology is found by minimizing the number of
substitutions needed to explain the informative sites. Parsimony
methods are not based on explicit evolutionary models.

**Maximum Likelihood.** Maximum likelihood and Bayesian methods
(see below) are probabilistic methods of inference. Both have the
pleasing properties of using explicit models of molecular evolution
and allowing for rigorous statistical inference. However, both
approaches are very computer intensive.

A stochastic model of molecular evolution is used to assign a probability (likelihood) to each phylogeny, given the sequence data of the OTUs. Maximum likelihood inference [Felsenstein, 1981] then consists of finding the tree which assign the highest probability to the data.

**Bayesian inference.** The objective of Bayesian phylogenetic
inference is not to infer a single "correct" phylogeny, but rather
to obtain the full posterior probability distribution of all
possible phylogenies. This is obtained by combining the likelihood
and the prior probability distribution of evolutionary parameters.
The vast number of possible trees means that bayesian phylogenetics
must be performed by approximative Monte Carlo based
methods.[Larget and Simon, 1999], [Yang and Rannala, 1997].