Genetic Differentiation and Clonality in a Local Population of the Caucasian Endemic Trifolium polyphyllum C.A. Mey. (Fabaceae)

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Abstract

Trifolium polyphyllum is a Caucasian endemic of the Fabaceae family peculiar for its inability of nitrogen fixation. Despite this unique trait, the species is insufficiently studied, in particular, little is known about its propagation and dispersal modes. Analyses of ISSR markers in a sample from a population at Malaya Khatipara Mountain revealed that the species is capable to both sexual and vegetative propagation; however, the former mode dominates. We found out that separate patches within a local population are considerably genetically differentiated within an area of about 2000 square meters (PhiPT = 0.349; p = 0.001). We suppose this may happen due to lack of adaptations to seed dispersal. We also suppose that the observed concentration of genetically admixed individuals in upper parts of slopes is due to peculiarities of pollinators’ behavior. The size of vegetative clones does not exceed 1 square meter.

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About the authors

O. B. Zelenova

Lomonosov Moscow State University

Author for correspondence.
Email: ischanzer@gmail.com
Russian Federation, Moscow

M. A. Galkina

Tsitsin Main Botanical Garden of Russian Academy of Sciences

Email: ischanzer@gmail.com
Russian Federation, Moscow

V. G. Onipchenko

Lomonosov Moscow State University

Email: ischanzer@gmail.com
Russian Federation, Moscow

I. A. Schanzer

Tsitsin Main Botanical Garden of Russian Academy of Sciences

Email: ischanzer@gmail.com
Russian Federation, Moscow

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2. Fig. 1. a – Position of the studied population in the Caucasus (red triangle), b – “cushion” of T. polyphyllum (photo by V. Onipchenko); c – location of sampling sites on Mount Malaya Khatipara: 1 – first transect, 2 – second transect, 3 – first “pillow”, 4 – second “pillow”. Transects were divided into rectangular areas of 1 × 0.5 m, from which samples were taken. The image of the mountain was obtained using the Google Earth service (https://earth.google.com/web/). Scale bar 60 m.

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3. Fig. 2. Results of the study of the first sample. a – Ordination using the NMDS method: 1 – first transect, 2 – second transect, 3 – first “pillow”, 4 – second “pillow”; b – clustering of samples of the first and second transects using the UPGMA method, Jaccard distance; c – clustering of samples of the first and second “pillows” using the UPGMA method, Jaccard distance. Bootstrap support values ​​above 60% are shown near the corresponding nodes. Samples from the first transect are indicated in red, from the second – in green; from the first “pillow” - blue, from the second “pillow” - crimson.

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4. Fig. 3. Results of the study of the first sample in the STRUCTURE program. a – Graph of ΔK; b – bar graph of posterior probabilities of assigning samples to genetic clusters for K = 4 and the location of samples on transects, circles indicate samples with a probability of assignment to the corresponding cluster >95%, triangles indicate genetically mixed samples, color corresponds to the cluster to which the sample belongs most probability, transects and the location of samples on them are depicted schematically; c – ordination using the NMDS method, sample designations are the same, numbers indicate cluster numbers, fractions indicate cluster numbers to which the sample belongs with the highest/lowest probability. Transects and the location of samples on them are shown schematically above the bar graph and are designated 1 and 2, respectively.

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5. Fig. 4. Results of analysis of samples of the second sample. a – Ordination by NMDS method, 1 – first transect, 2 – second transect; b – clustering of samples from the first and second transects using the UPGMA method, Jaccard distance; samples from the first transect are indicated in red, from the second – in green.

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