Mitochondrial DNA Data

Mitochondrial DNA sequences largely confirmed the results from allozyme studies, also indicating minimal within-population variation and the separation of the South Slovenian samples. We amplified a fragment of the cytochrome oxidase I (COI)

gene using primers C1-J-2183 (Jerry) and TL2-N-3014 (Pat) (Simon et al. 1994) and successfully sequenced 608 bp for 114 specimens of C. v. nodulosus and six individuals from three Romanian populations of C. v. variolosus. Only nine different haplotypes were identified (Fig. 3). There was hardly any haplotype variation within populations of C. v. nodulosus. With the exception of two individuals deviating by one and three nucleotide changes, respectively, all remaining 92 individuals

Fig. 3 Neighbor-joining tree (Saitou and Nei 1987) based on the number of nucleotide changes between 120 sequences of COI fragment. Numbers at nodes refer to support from 1,000 bootstrap replicates. Central European and Southern Slovenian specimens: C. v. nodulosus, Romanian specimens: C. v. variolosus. Phylogenetic analyses were conducted in MEGA4 (Tamura et al. 2007)

Fig. 3 Neighbor-joining tree (Saitou and Nei 1987) based on the number of nucleotide changes between 120 sequences of COI fragment. Numbers at nodes refer to support from 1,000 bootstrap replicates. Central European and Southern Slovenian specimens: C. v. nodulosus, Romanian specimens: C. v. variolosus. Phylogenetic analyses were conducted in MEGA4 (Tamura et al. 2007)

in the Central European populations (1-10) shared the same haplotype. The 20 individuals from Southern Slovenian (populations 11 and 12) shared a single haplotype that differed from the former by four nucleotide changes. All of these were separated from C. v. variolosus haplotypes by at least four nucleotides (Fig. 3). This finding corroborates that the Southern Slovenian populations of C. v. nodulosus survived in a different glacial refugium than the remaining European samples (cf. Chap. 5.1, this contribution).

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