Phylogenetic Analyses
The nexus format of the aligned data matrix was imported to phylo-
genetic analysis software PAUP, and executed for the sequence
data analyses. Strict consensus tree and bootstrap strict consensus
tree [Fig-4], [Fig-5] resulted on completion of the analyses were
saved for presentation. In addition with parsimony analyses, NJ
analysis were also performed using PAUP to infer NJ tree [Fig-5]
based on branch length.
The parsimony analysis (using PAUP) of the entire ITS region re-
sulted in 1280 maximally parsimonious trees (MPTs) with a total
length of 480 steps, a consistency index (CI) of 0.754 (0.730 CI
excluding uninformative characters), a homoplasy index (HI) of
0.456 (0.432 HI excluding uninformative characters), rescaled con-
sistency index (RC) of 0.579 and a retention index (RI) of 0.767.
One of the MPTs has been shown in [Fig-5] in which number above
the lines indicate the bootstrap support in 100 replicates.
A perusal of all trees (Strict Consensus Tree, Bootstrap Strict Con-
sensus Tree, and NJ tree) clearly indicates that Limonium carno-
sum included in this study from geographic region Saudi Arabia
phylogenetically very closely related with Limonium narbonense
and Limonium vulgare. It is also clearly evident that Limonium cylin-
drifolium and Limonium axillare does not nested deeply within the
phylogenetic tree, rather were found at the base of the phylogenetic
tree and L. lobatum occupies most basal position in phylogenetic
tree. Limonium cylindrifolium and Limonium axillare shows sister
relationships; In ITS sequences both these two species differs in 5
base pairs in ITS2 region. Therefore, ITS2 secondary structure for
these two species, and additionally all taxon included in the analysis
were predicted to bring molecular morphological signature of the
Limonium distributed in Saudi Arabia.
Average Evolutionary Divergence over all Sequence Pairs was
estimated. The number of base substitutions per site from averag-
ing over all sequence pairs was 0.129. The result was based on the
pairwise analysis of 24 sequences included in the analyses. Anal-
yses were conducted using the Maximum Composite Likelihood
method in MEGA4 [46]. Maximum Composite Likelihood Estimate
of the Pattern of Nucleotide Substitution [Table-7] was performed
using MEGA4 [46].
Table 7- Maximum Composite Likelihood Estimate of the Pattern of
Nucleotide Substitution
Each entry shows the probability of substitution from one base (row) to
another base (column) instantaneously.
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