Differentiation and Taxonomic Identification of Roburoid Oaks in the Caucasian and Crimean Regions Using Nuclear Microsatellite Markers

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Abstract

The inter- and intra-specific structure of genetic variability was studied using 18 microsatellite loci (nSSRs) in closely related roburoid oaks in the Crimean-Caucasian region.The seven most widespread Quercus taxa in the region were studied in 29 morphologically pure populations from different parts of the North Caucasus, Transcaucasia, Crimea and northeastern Europe. Most taxa were studied using nSSR markers for the first time. Among the 492 trees studied, Bayesian clustering method implemented in STRUCTURE identified clusters corresponding to the pedunculate oak Quercus robur, the Hartwiss oak Q. hartwissiana, the Caucasian oak Q. macranthera, the downy oak Q. pubescens and three subspecies of sessile oak: Q. petraea ssp. petraea, Q. petraea ssp. iberica, Q. petraea ssp. medwediewii. Geographic structure was identified within Q. robur, Q. pubescens and Q. p. ssp. petraea. The 18 nSSR loci used are efficient in the taxonomic assignment of individuals, and identifying hybrids. The close relationship between the “long-pedunculate” roburoid oaks (Q. robur and Q. hartwissiana) is shown, with a greater difference from other species. For one of the subspecies of sessile oak, widespread in the North Caucasus and Crimea Q. petraea ssp. medwediewii (syn. Q. calcarea), or limestone oak, significant differences from other taxa were found, reaching the species level. The assumption of a possible hybrid origin of this taxon as a result of hybridization of Q. petraea and Q. pubescens is not confirmed by genetic analysis. The other two subspecies of Q. petraea (Q. p. ssp. petraea and Georgian oak Q. p. ssp. iberica) are differentiated to a lesser extent and are related to each other, which confirms the legitimacy of distinguishing two geographically isolated taxa at the rank of subspecies. The highest variability was observed in Q. pubescens (He = 0.777). In Q. p. ssp. medwediewii variability was lower than in other widespread taxa (He = 0.652), and was approximately at the level of variability of Q. hartwissiana (He = 0.633) and Q. macranthera (He = 0.659). Clear differentiation of taxa by nuclear markers shows the limited introgression in closely related oak species in the Caucasus and Crimea. The identified genetic clusters can be used as reference groups for further population genetic studies of oaks in the Crimean-Caucasian region.

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S. A. Semerikova

Institute of Plant and Animal Ecology, Ural Branch of Russian Academy of Science

Author for correspondence.
Email: s.a.semerikova@ipae.uran.ru
Russian Federation, Ekaterinburg, 620144

Kh. U. Aliev

Mountain Botanical Garden of Dagestan Federal Research Centre of the Russian Academy of Sciences

Email: s.a.semerikova@ipae.uran.ru
Russian Federation, Makhachkala, 367000

V. L. Semerikov

Institute of Plant and Animal Ecology, Ural Branch of Russian Academy of Science

Email: s.a.semerikova@ipae.uran.ru
Russian Federation, Ekaterinburg, 620144

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2. Fig. 1, a. Geographic distribution of the studied samples of seven taxa of roburoid oaks in the Crimean-Caucasian and northern European parts of the range. The sample numbers correspond to those indicated in Table 1. Taxon designations: Rb – Q. robur, Pb – Q. pubescens, Mc – Q. macranthera, Htw – Q. hartwissiana, Pt_pt – Q. petraea ssp. petraea, Pt_ib – Q. petraea ssp. iberica, Pt_med – Q. petraea ssp. medwediewii. The color designations of the samples correspond to the predominant genetic clusters STRUCTURE at K = 6 (Fig. 1, b). The dotted line indicates the northern boundary of the oak range in the region.

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3. Fig. 1, b. Distribution of genetic clusters calculated using STRUCTURE based on 18 microsatellite loci for population 29 (492 individuals) of five Quercus species with K = 2.6. Each sample is represented by a vertical column divided into two or six colored segments proportional to the contribution of each genetic cluster.

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4. Fig. 2. PCoA ordination based on the variability of 18 nSSR loci. a – populations of seven taxa of roburoid oaks; b – populations of sessile oaks Q. petraea s. l., Q. macranthera, Q. pubescens. Populations are designated by color in accordance with the dominant cluster STRUCTURE with the number of clusters K = 6 for the set 1 (see Fig. 1, b).

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5. Fig. 3. Distribution of genetic clusters calculated using STRUCTURE based on 18 microsatellite loci for 364 individuals of Q. petraea s. l., Q. macranthera, Q. pubescens (set 2) with K = 4, 5, 6, 7. Each sample is represented by a vertical column divided into segments proportional to the participation of each genetic cluster.

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