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Relict populations of Quercus calliprinos Webb on Sardinia island identified by chloroplast DNA sequences

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Among species of the genus Quercus L. in temperate regions, the horny oak is indigenous in Southern Europe and from Northern Africa to the Middle East. Several authors have classified the "horny oak complex" in different way, taking into account the variability of different morphological traits. Horny oak shows great polymorphism, and the exact geographic distribution of the populations is not easily defined. Moreover, another complication is the unclear systematics, and some authors suggest that the horny oak complex split in two different species: Quercus calliprinos Webb, which has an eastern distribution (the Mediterranean basin), and Quercus coccifera L., which has a more western distribution. To contribute to the systematics of three relict populations of horny oak on Sardinia, we studied the trnL-trnF region of chloroplast DNA (cpDNA). We evaluated eleven horny oak populations sampled throughout the geographic distribution of the species, as well as six additional species of Quercus and Fagus sylvatica L. for comparison. In the Fagaceae family, the variable and informative sites at the interspecific level are localised in the sequences of the introns, whereas the intergenic region has a higher variability at the intraspecific level, indicating that the intergenic region evolves at a faster mutation rate than the intron. It was possible to distinguish between the different species of Fagaceae and the two systematic entities of horny oaks studied with cpDNA analysis. Moreover, it was possible to identify the relict horny oak populations on Sardinia as Q. calliprinos. These populations represent an isolated group of Q. calliprinos in the western Mediterranean basin. The geographic isolation permitted a mutation to arise in the intergenic region near a microsatellite, and this mutation was fixed in the group and identifies the relict Sardinia populations.
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... The effectiveness of this region in distinguishing the systematic entities in Quercus spp. has been previously demonstrated (PAFFETTI et al., 2001). This cpDNA region is constituted by the trnL intron, the second trnL exon, and the tmL-trnF intergenic spacer. ...
... Amplifications were performed in a 20 pl volume as reported by PAFFETTI et al. (2001) using the universal primers (c and d) for the tmL-tmF cpDNA region described by TABERLET et al. (1991). A Perkin Elmer 9700 thermocycler was used. ...
... The intergenic region had a higher variability at the intraspecific level. These data confirm the results of previous studies, conducted in the genus Quercus, in the potential of this sequence in phylogenetic study (PAFFETTI et al. 2001). The trees obtained with the different methods of phylogenetic tree reconstruction (parsimony, and maximum-likelihood) show two separated clusters of the Fagus spp., one clustering with I;: grandijolia and the other with I;: sylvatica. ...
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... The geographic distribution of the two entities is not clear [Paffetti et al., 2001]. A. Camus [1936Camus [ -1938 has indicated that the western Mediterranean basin is the distribution range for Q. coccifera and the eastern zone for Q. calliprinos. ...
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