The study presents the results of the modern phylogeographic structure of Anemone baicalensis Turcz. on the Khamar-Daban Ridge using trnL-trnF plastid DNA marker. Being one of nemoral relict species with the longest ranges along the Khamar-Daban Ridge, A. baicalensis is a convenient model species to study the possible isolation of relict species populations in microrefugia during the last Pleistocene glaciation, followed by their recolonization during subsequent climate change. The complete trnL-trnF region was amplified using GoTaq Flexi DNA Polymerase (Promega) and the universal primers for the flank regions of trnL and trnF plastid DNA genes. Gel-purified amplicons were sequenced by the Sanger method using 3500 Genetic Analyzer. The multiple alignments of nucleotide sequences were prepared by the MUSCLE algorithm followed by manual editing. The indels were considered as evolutionary events and included as a separate binary data partition to the end of matrix. Phylogenetic reconstructions were performed by both the maximum likelihood and Bayesian inference methods. Three trnL-trnF haplotypes were revealed in A. baicalensis populations on the Khamar-Daban Ridge. The first haplotype was typical for the populations from the eastern part of the range, two others – for populations from the western part. The haplotypes found in the populations from the western part of the range formed a well-supported haplogroup according Bayesian inference analysis. The described phylogeographic patterns confirm the partial fragmentation of A. baicalensis range and preservation of populations in microrefugia on the Khamar-Daban Ridge during the latest Pleistocene cooling. The zone of secondary contact and possible hybridization of the populations from the different phylogeographic groups appeared after the onset of favorable conditions is located on the site between the Bolshoi Mamai and Vydrinaya rivers, where both ‘eastern’ and ‘western’ haplotype variants were revealed. The presence of a common haplotype in populations in the eastern border (the Bolshaya Rechka river) and the central part of the range suggests that the geographical isolation of eastern populations is currently not absolute and they can contact with the main part of the range in mountain taiga and subalpine zones.

Protopopova Marina Vladimirovna, Candidate of Science (Biology), Senior Research Scientist, Siberian Institute of Plant Physiology and Biochemistry SB RAS, 132, Lermontov st., Irkutsk, 664033, Russian Federation; Associate Professor, Irkutsk State University, 1, K. Marx st., Irkutsk, 664003, Russian Federation, e-mail: marina.v.protopopova@gmail.com 

Pavlichenko Vasiliy Valeryevich, Candidate of Science (Biology), Senior Research Scientist, Siberian Institute of Plant Physiology and Biochemistry SB RAS, 132, Lermontov st., Irkutsk, 664033, Russian Federation; Associate Professor, Irkutsk State University, 1, Karl Marx st., Irkutsk, 664003, Russian Federation, e-mail: vpavlichenko@gmail.com 

Orlova Daria Alekseevna, Student, Irkutsk State University, 1, K. Marx st., Irkutsk, 664003, Russian Federation; Engineer, Siberian Institute of Plant Physiology and Biochemistry SB RAS, 132, Lermontov st., Irkutsk, 664033, Russian Federation 

Chepinoga Victor Vladimirovich, Doctor of Sciences (Biology), Leading Research Scientist, V. B. Sochava Institute of Geography SB RAS, 1, Ulan-Batorskaya st., Irkutsk, 664033, Russian Federation; Professor, Irkutsk State University, 1, K. Marx st., Irkutsk, 664003, Russian Federation, e-mail: victor.chepinoga@gmail.com

Protopopova M.V., Pavlichenko V.V., Orlova D.A., Chepinoga V.V. Phylogeographic Structure of Anemone baicalensis (Ranunculaceae) Based on Plastid DNA Polymorphism (trnL-trnF) as an Evidence of Pleistocene Microrefugia Existence on the Khamar-Daban Ridge (Southern Baikal Region). The Bulletin of Irkutsk State University. Series Biology. Ecology, 2019, vol. 30, pp. 3-15. https://doi.org/10.26516/2073-3372.2019.30.3 (in Russian)

nemoral relict, refugium, glaciation, molecular genetic markers, techniques of molecular biolo-gy, trnL-trnF, Anemone baicalensis

Ivanovsky L.N. Psevdolednikovye formy rel'efa v doline r. Vydrinoi [Pseudo-glacial landforms in the valley of the Vydrina river (Southern Baikal region)]. Geography and natural resources, 2006, no. 4, pp. 161-167. (in Russian)

Iljin M.M. Tretichnye reliktovye elementy v taezhnoi flore Sibiri i ikh vozmozhnoe proiskhozhdenie [The Tertiary relict elements in the Siberian Taiga flora and their possible origin]. Materialy po istorii flory i rastitel'nosti SSSR [Materials on the history of the flora and vegetation of the USSR], is. 1. Moscow-St.-Petersb., AS USSR Publ., 1941, pp. 257-292. (in Russian)

Protopopova M.V., Pavlichenko V.V., Gnutikov A.A., Adelshin R.V., Chepinoga V.V. Ispol'zovanie geneticheskikh markerov dlya otsenki sostoyaniya reliktovykh vidov rastenii Baikal'skoi Sibiri [Application of genetic markers for ecological status assessment of the relict plant species of Baikal Siberia]. Bull. RUDN Univ. Series Ecology and life safety, 2015, no. 4, pp. 28-36. (in Russian)

Krasnaya kniga Krasnoyarskogo kraya. Redkie i nakhodyashchiesya pod ugrozoi ischeznoveniya vidy dikorastushchikh rasteniĭ i gribov [Red Book of the Krasnoyarsk territory. The Rare and Endangered Species of Wild Plants and Fungi]. Krasnoyarsk, Siberian Fed. Univ. Publ., 2012, 576 p. (in Russian)

Krasnaya kniga Respubliki Buryatiya: redkie i nakhodyashchiesya pod ugrozoi ischeznoveniya vidy zhivotnykh, rastenii i gribov [Red Data Book of Republic of Buryatia: Rare and Endangered Species of Animals, Plants and Fungi]. Ulan-Ude, Buryat SC SB RAS Publ., 2013, 688 p. (in Russian)

Chepinoga V.V., Mishina A.V., Protopopova M.V., Pavlichenko V.V., Bystrov S.O., Vilor M.A. Novye dannye o rasprostranenii nekotorykh nemoral'nykh reliktovykh rastenii v predgor'yakh khrebta Khamar-Daban (Yuzhnoe Pribaikal'e) [New data on distribution of several nemoral relict plant species on the foothills of the Khamar-Daban ridge (Southern Baikal)]. Botanicheskii Zhurnal, 2015, vol. 100, no. 5, pp. 478-489. (in Russian)

Protopopova M.V., Pavlichenko V.V., Konovalov A.D., Zolotovskaya E.D., Bairamova E.M., Chepinoga V.V. Perspektivy ispol'zovaniya vnutrennikh transkribiruemykh speiserov (ITS1 i ITS2) dlya identifikatsii redkikh vidov rastenii na primere roda Waldsteinia (Rosaceae) [Perspectives for the Internal Transcribed Spacer (ITS1 and ITS2) application for the endangered plant species identification with Waldsteinia (Rosaceae) as an example]. Bull. Irkutsk St. Univ. Series Biology. Ecology, 2016, vol. 17, pp. 5-11. (in Russian)

Polozhii A.V., Krapivkina E.D. Relikty tretichnykh shirokolistvennykh lesov vo flore Sibiri [Relicts of Tertiary Broad-Leaved Forests in Siberian Flora]. Tomsk, Tomsk St. Univ. Publ., 1985, 158 p. (in Russian)

Belov A.V., Bezrukova E.V., Sokolova L.P., Abzayeva A.A., Letunova P.P., Fisher E.E., Orlova L.A. Rastitelnost Pribaikal'ya kak indikator global'nykh i regionalnykh izmeneniĭ prirodnykh usloviĭ severnoi Azii v pozdnem kainozoe [Vegetation of the Baikal region as an indicator of global and regional changes in natural conditions of North Asia in the late Cainozoic]. Geography and Natural Resources, 2006, no. 6, pp. 5-18. (in Russian)

Krestov P.V., Barkalov V.Y., Omelko A.M., Yakubov V.V., Nakamura Y., Sato K. Reliktovye kompleksy rastitel'nosti sovremennykh refugiumov severo-vostochnoi Azii [Relic vegetation complexes in the modern refugia of Northeast Asia]. Komarovskie Chtenia [V.L.Komarov Memorial Lectures], 2009, vol. 56, pp. 5-63. 

Chepinoga V.V., Protopopova M.V., Pavlichenko V.V. Detection of the most probable pleistocene microrefugia on the northern macroslope of the Khamar-Daban Ridge (Southern Prebaikalia). Contemporary Problems of Ecology, 2017, vol. 10, no. 1, pp. 38-42. https://doi.org/10.1134/S1995425517010036

Protopopova M., Pavlichenko V., Gnutikov A., Chepinoga V. DNA barcoding of Waldsteinia Willd. (Rosaceae) species based on ITS and trnH-psbA nucleotide sequences. Information Technologies in the Research of Biodiversity. Springer Proc. Earth Environ. Sci., Springer Nature Switzerland, 2019, pp. 107-115. https://doi.org/10.1007/978-3-030-11720-7_15

Doyle J.J., Doyle J.L. A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem. Bull., 1987, vol. 19, pp. 11-15.

Felsenstein J. Evolutionary trees from DNA sequences: a maximum likelihood approach. J. Mol. Evol., 1981, vol. 17, no. 6, pp. 368-376.

Gnutikov A.A., Protopopova M.V., Pavlichenko V.V., Chepinoga V.V. IAPT chromosome data 28. Taxon, 2018, vol. 67, no. 6, pp. 1236-1237. https://doi.org/10.12705/676.39

Gnutikov A.A., Protopopova M.V., Pavlichenko V.V., Chepinoga V.V. IAPT/IOPB chromosome data 22, Taxon, 2016, vol. 65, no. 5, pp. 1201. http://dx.doi.org/10.12705/655.40

Gnutikov A.A., Protopopova M.V., Chepinoga V.V., Konovalov A.D., Zolotovskaya E.D., Pavlichenko V.V. IAPT/IOPB chromosome data 26. Taxon, 2017, vol. 66, no. 6, pp. 1488-1489. https://doi.org/10.12705/666.30

Kumar S., Stecher G., Tamura K. MEGA7: Molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol. Biol. Evol., 2016, vol. 33, no. 7, pp. 1870-1874. https://doi.org/10.1093/molbev/msw054

Mosyakin S.L. Further new combinations in Anemonastrum (Ranunculaceae) for Asian and North American taxa. Phytoneuron, 2018, vol. 55, pp. 1-11.

Tamura K. Estimation of the number of nucleotide substitutions when there are strong transition-transversion and G+C-content bases. Mol. Biol. Evol., 1992, vol. 9, pp. 678-687.

Taberlet P., Gielly L., Pautou G., Bouvet J. Universal primers for amplification of three non-coding regions of chloroplast DNA. Plant Mol. Biol., 1991, vol. 17, no. 5, pp. 1105-1109.