Identification of cryptic forms of the hybridogenic complex of European water frogs (Pelophylax esculentus complex) in the conditions of transformed biotopes of the south of the Central Russian Upland based on DNA markers

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Abstract

AIM: The aim of the study was to carry out species identification of the hybridogenic complex of European water frogs (Pelophylax esculentus complex) in the conditions of transformed biotopes of the south of the Central Russian Upland based on molecular genetic markers

MATERIALS AND METHODS: The study included 36 local populations (770 individuals) living in conditions of transformed biotopes of the south of the Central Russian Upland. Identification of cryptic forms was carried out by a Multiplex PCR. Two molecular genetic markers were used for amplification: intron 1 of the SAI-1 DNA serum albumin gene to determine hybrids and cryptic forms, and a fragment of the first subunit of the cytochrome oxidase COI mtDNA gene to determine maternal lines.

RESULTS: According to the data obtained, “pure” R-type population systems predominate (58.33%) in the study region. Mixed RE-type population systems were identified in 14 localities, REL-type in the region is extremely rare and is noted only in one locality. No pure L-type, E-type, or mixed LE-type population systems have been identified. The study revealed a statistically significant (p < 0.001) predominance of haplotypes of the “Western” form (Pelophylax ridibundus).

CONCLUSIONS: The data obtained indicate active adaptive changes in the population structure of European water frogs in the study area. Degradation of water bodies caused by abiotic and anthropogenic factors forces amphibians to migrate to neighboring reservoirs in which hybridization of representatives of this complex occurs. Introgressive and hybrid forms of the marsh frog, as well as hybrid edible individuals with greater ecological plasticity and tolerance to anthropogenic pressure, displace populations of Pelophylax lessonae. Based on the above, we consider it necessary to include the pond frog (P. lessonae) in the Red Book of the Belgorod region.

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About the authors

Anatoliy S. Barkhatov

Belgorod State National Research University

Author for correspondence.
Email: barkhatov@bsu.edu.ru
ORCID iD: 0000-0001-9996-7251
SPIN-code: 3833-2940

Junior Research Associate, Research Center of Genomic Selection

Russian Federation, Belgorod

Eduard A. Snegin

Belgorod State National Research University

Email: snegin@bsu.edu.ru
ORCID iD: 0000-0002-7574-6910
SPIN-code: 5655-7828

Dr. Sci. (Biol.), Director of the Research Center of Genomic Selection

Russian Federation, Belgorod

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Supplementary files

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2. Fig. 1. Collection points. The numbering corresponds to Table 1

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3. Fig. 2. Fragment of an electrophoregram of multiplex PCR of genes. Row No. 1: SAI-1 YADNA. Wells 1, 4 — Pelophylax cf. bedriagae (the “eastern” form of the marsh frog); 2, 3, 14, 17 — P. ridibundus (the “western” form of the marsh frog); 5, 7, 13, 15, 16, 18 — P. esculentus (parent species P. lessonae and P. ridibundus, “western” form); 8 — P. esculentus (parent species P. lessonae and P. cf. bedriagae, “eastern” form); 10 — hybrid form of P. ridibundus and P. cf. bedriagae; 11 — DNA length marker 50 + bp; 12 — Pelophylax lessonae (pond frog). Row No. 2: mtDNA COI. Wells 1–4, 6–10, 12–18 — haplotypes of P. ridibundus; 5 — haplotype of P. cf. bedriagae; 11 — is a marker of DNA lengths of 50 + bp

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4. Fig. 3. Dried-up reservoir, Krasnogvardeysky District, Belgorod Region

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5. Fig. 4. Lake Bannoye, point No. 11 “Grafovka”

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