GENETIC HETEROGENEITY OF THE MARSH FROG
(PELOPHYLAX (RIDIBUNDUS) COMPLEX; ANURA, RANIDAE)
POPULATION IN CRIMEA REVEALED BY MITOCHONDRIAL AND NUCLEAR DNA ANALYSES 

Kukushkin Oleg Vital'evich, Researcher, department of biodiversity studies and ecological monitoring, T. I. Vyazemsky Karadag Research Station – RAS Nature Reserve (24 Nauki street, Kurortnoe settlement, Theodosia, Russia); applicant, Department of Herpetology, Zoological Institute of RAS (1 Universitetskaya embankment, Saint-Petersburg, Russia), 
E-mail: mtasketi2018@gmail.com
Ivanov Aleksandr Yur'evich, Teacher, sub-departmnet of zoology and ecology, Penza State University (40 Krasnaya street, Penza, Russia), E-mail: akella58@mail.ru
Ermakov Oleg Aleksandrovich, Candidate of biological sciences, associate professor, sub-departmnet of zoology and ecology, Penza State University (40 Krasnaya street, Penza, Russia), E-mail: oaermakov@list.ru 

597.851 (477.75) 

DOI

10.21685/2307-9150-2018-3-3 

Background. According to current concept, the marsh frog, Pelophylax ridibundus (Pallas, 1771), is a diverse complex of cryptic species and evolutionary lineages with an unelucidated taxonomical status. The low level of reproductive isolation beetwen the forms of P. (ridibundus) complex defines their hybridizing potential, and besides the introgression zones could embrace quite large areas. The objectives of the current study was to dissect the genetic structure and features of spatialbiotopic distribution of “western” and “eastern” forms of a marsh frog (the middle-European lineage P. ridibundus s. l. and the Anatolian lineage P. cf. bedriagae s. str.,
respectively) throughout the Crimea.
Materials and methods. Molecular analysis of 188 individuals of a marsh frog from 55 sites at the Crimean Peninsula was based on two genetic markers: a fragment of the first subunit of cytochrome oxydase (COI) mtDNA gene, and the first intron of the serum albumin (SAI-1) nuDNA gene. Differences in allele frequencies in different landscape areas of the peninsula (Plain Crimea, Piedmont area, Main Range, Southern Coast) were assessed using Pearcon’s chi-squared (χ2) and Fisher’s two-tailed tests. The analysis of the biotopic distribution of the of the marsh frog populations with one and two mitotypes was performed with considering of an altitude range above the sea level, water body type and location, vegetation type as well as the degree of anthropogenic disturbance.
Results. MtDNA marker-based analysis revealed the presence of “eastern” form specific mitotype in 87 % of marsh frog individuals, while the “western” form comprised only 13 %. In turn, nuDNA marker-based analysis showed that the frequency of alleles specific for P. cf. bedriagae and P. ridibundus was in 4:1 ratio. Frogs with mtDNA marker specific for the “western” form reside predominantly on northern macroslope of the Main Range, where this mitotype was found in almost 1/5 of the total sample of the assessed individuals, while frogs belonging to the “western” form are mainly confined to the Chernaya River basin in the south-west of the Crimean Peninsula and the pre-yaila (highland) terrains of the central-eastern part of the Main Range. It is noteworthy that the populations including individuals characterized by the “western” form mitotype prefer mostly undisturbed wooded habitats – the gorges of mountain rivers, cold-water springs and lakes under the forest canopy.
Conclusions. The discordant mt- and nuDNA markers distribution patterns in the Crimea along with the data on “western” form habitat distribution suggests a complex colonization history of the peninsula by representatives of P. (ridibundus) complex. Apparently, the hybridization of “western” and “eastern” forms of a marsh frog occured throughout the Crimea. Thus, the “western” form should be considered as a late Pleistocene relic, because its distribution in the Crimean Peninsula is restricted and associated with most isolated mountains areas.

Key words

Pelophylax ridibundus, Pelophylax cf. bedriagae, cytochrome oxydase mtDNA gene, serum albumin nuDNA gene, the Crimean Peninsula, introgressive hybridization, relics, Pleistocene, Holocene 

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