Bayesian phylogenetic analysis in BEAST [36] of all unique mainland and
island haplotypes (Fig. 3) used the earliest island fox radiocarbon date as a prior estimate for the coalescence of all island fox lineages.
Santa Catalina and Santa Cruz are the only islands that share a haplotype, which is more closely related to the northern
island haplotypes than the southern.
Not exact matches
All 10 of the novel
haplotypes were found on the outer Scottish
islands.
The only locality with both
haplotypes was in the Solomon
Islands.
The mitogenome sequences revealed a total of 35
haplotypes with 14 found exclusively on the
islands and 21 found only in mainland California.
All of the
islands had a least one private
haplotype, but 19 of 41 Santa Catalina foxes (southern
island) shared a
haplotype with Santa Cruz foxes (northern
island).
The northern
islands had nine closely related
haplotypes while the southern
islands had five
haplotypes that were more distant from each other (Fig. 2).
Haplotype and nucleotide diversity in the
island populations was markedly lower than in the mainland populations, with only one to five
haplotypes per
island.
No other
islands shared a
haplotype, although San Clemente and San Nicolas each had two
haplotypes that were separated by a single base pair.
A positive correlation was identified between
island area and the number of
haplotypes recovered and
haplotype diversity (Pearson's r = 0.80 p - value = 0.03 and r = 0.77 p - value = 0.04, respectively).
Remarkably,
island fox
haplotypes formed a monophyletic clade nested within clade B, rather than with foxes from southern California, closest to the Channel
Islands.
Among the gray foxes sampled, the
haplotypes that were more closely related to the
island fox were from Lassen / Shasta counties in northern California.
The aligned mitogenomes were visually examined and when a single
island individual or an ambiguous base generated a unique
haplotype, Sanger sequencing was conducted to verify the basecalls (S1 Text and S4 Table), which did not change except for two samples with conflicting
haplotypes that was discarded from all analyses.
We identified two pairs of closely related
haplotypes on each of the two most remote
islands in the archipelago (San Nicolas and San Clemente), which indicates in situ evolution of
island fox genetic variation.
This pattern is strikingly different from the southern
islands, where San Clemente and San Nicolas each have two
haplotypes just a single base pair away from each other.
Island foxes show greater mitogenome diversity in the northern
islands with nine
haplotypes that differ from 2 — 5 base pairs from the central node of the star - shaped radiation (Fig. 2).
Previous analyses using mtDNA RFLP's identified more mtDNA
haplotypes than detected by our whole mitogenome (WMG) sequencing analysis on San Miguel (2 RFLP: 1 WMG genotypes) and Santa Catalina (3 RFLP: 2 WMG genotypes), suggesting a possible loss of mtDNA diversity on these
islands as a result of population bottlenecks over the past 25 years [15].