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Salvia officinalis survived in situ Pleistocene glaciation in ‘refugia within refugia’ as inferred from AFLP markers


Salvia officinalis is a perennial species, native and endemic to the Western Balkans and the Apennine Peninsula. Due to its medicinal and aromatic properties, it is used in pharmaceutical, cosmetic, and food industries. The main objectives of the study were to infer the genetic structure of S. officinalis populations in the northern and central parts of the eastern Adriatic coast, to detect the phylogeographical barriers among the putative microrefugia and to assess the genetic diversity among the resulting ancestral clusters. Twenty-five populations were assessed using amplified fragment length polymorphism markers. High polymorphism and high diversity within populations were typical for this outcrossing long-lived species. The Fitch–Margoliash tree based on Nei’s genetic distance matrix showed that most of the populations tended to group in accordance with the geographical position of their collecting sites. Spatial analysis of the genetic diversity revealed a typical pattern of isolation by distance. Very low overall among-population differentiation and detection of only three private alleles indicate that there has been high gene flow among populations. By using Bayesian Analysis of Population Structure on population level, two distinct ancestral clusters were obtained. It is likely that these two ancestral clusters were separated for a longer period by Pleistocene glaciation, although the subsequent fast recolonization resulted in diminished genetic differences. High rarity of northern and southern populations of the investigated area indicates that S. officinalis presumably survived in both northern and southern microrefugia and expanded from there resulting in secondary contact zones, characterized by lower rarity and equal genetic diversity.

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This research has been funded by the Project KK. Biodiversity and Molecular Plant Breeding, Centre of Excellence for Biodiversity and Molecular Plant Breeding (CoE CroP-BioDiv), Zagreb, Croatia.

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Conceived and designed the experiments: Zlatko Šatović, Zlatko Liber, and Marija Jug-Dujaković. Performed the experiments: Marija Jug-Dujaković and Zlatko Liber. Analyzed the data: Zlatko Šatović. Contributed reagents/materials/analysis tools: Zlatko Šatović, Marija Jug-Dujaković, and Zlatko Liber. Contributed to the writing of the manuscript: Marija Jug-Dujaković, Tonka Ninčević, Zlatko Šatović, Zlatko Liber, and Martina Grdiša. All the authors have read and approved the final version of the manuscript.

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Correspondence to Tonka Ninčević.

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The authors declare that they have no conflict of interest.

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Collection of cuttings from protected areas was permitted by the authority of Directorate for Nature Protection of the Ministry of Culture of the Republic of Croatia (532-08-02-1/8-06-2; 20.04.2006) and from Javna ustanova Kamenjak (005/2006; 25.05.2006). For other locations, no specific permits were required because cuttings collection did not involve endangered or protected species.

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Handling Editor: Stanislav Španiel.

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Contribution to “Plants of the Balkan Peninsula in Space and Time.”

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Jug-Dujaković, M., Ninčević, T., Liber, Z. et al. Salvia officinalis survived in situ Pleistocene glaciation in ‘refugia within refugia’ as inferred from AFLP markers. Plant Syst Evol 306, 38 (2020).

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  • AFLP
  • Balkan Peninsula
  • Glaciation
  • Pleistocene
  • Refugium
  • Salvia officinalis