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Finally, we examine the hypothesis of isolation, which if accepted would discredit the human-mediated dispersal hypothesis.
Mantel's test results found significant correlation between Fst values and geographic distance (p = 0.028) corroborating the hypothesis of isolation by distance suggested by intron + HEG network.
To test a hypothesis of isolation by distance, geographical distances (km) among pairs of S. australis population samples were linearly regressed against their pairwise FST estimates.
Following the recommendations of Rousset (1997), the hypothesis of isolation by distance was tested by calculating the correlation between the matrices representing pairwise Slatkin genetic distances FST/(1− FST; Slatkin 1995) and the natural logarithm of geographic distance for each pair of P. infestans populations.
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To verify the hypothesis of isolation-by-distance (IBD), correlation between pair-wise linearized Φst/ 1-Φst) and river distances (Km) between populations were analyzed using the Mantel test [ 83] with 10,000 permutations.
The same program was used to run a Mantel test (Mantel 1967), with 10 000 permutations, to estimate the correlation of pairwise linearized distances with a matrix of linear geographic distances to test the hypothesis of genetic isolation by geographic distance (isolation by distance, IBD).
This lends strong support to the hypothesis of long isolation of the Spanish populations and independent evolution of snail ecotypes in Spain and in the two other regions.
While mtDNA and microsatellite divergence estimates reported above support the hypothesis of considerable isolation and differentiation between guppies inhabiting streams on the opposite sides of the Cordillera de la Costa, STRUCTURE results do not corroborate such a clear distinction.
Distances between sites were designed to account for a null hypothesis of genetic isolation by geographic distance, with gradients representing three geographic scales termed microgeographic (<2 km), mesogeographic (>2 km, <120 km), and macrogeographic (>120 km).
To test whether the degree of relatedness between individuals depends on the geographical distance, we used a procedure implemented in SPAGeDI which performs permutations of individual locations to determine the relationship between genetic ancestry and spatial distance expected under the null hypothesis of no isolation by distance.
In agreement with the hypothesis for isolation of inland populations, we found lower levels of genetic differentiation, as measured by Nei's DA, FST and standardized FST, in coastal sites and very high levels of genetic differentiation including the fixation of different alleles in many inland populations.
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