GENETIC FEATURES OF TARPAN-LIKE HORSES OF THE POLISH PRIMITIVE (THE KONIK) BREED AND ESTABLISHMENT OF PHYLOGENETIC OF ANCIENT EGUIDS USING THE ISSR-PCR MARK
Authors: Dzitsuik V. , Starodub L. , Dyman T. 
Published: March 2023

In the development of human society, the horse, compared to other domestic animals, played a central role. It became an integral part of the economic and intellectual development of ancient human settlements, a symbol of the transformation of subsequent great epochs. The last wild horse that survived in Ukraine until the end of the XIX century, according to many paleontologists, was the tarpan. However, this statement is still questioned.

In the books and documents of old chroniclers and researchers we find descriptions of wild horses called tarpans, which could be found in the wooded areas of Poland, Lithuania and Prussia in the XVIII century. Emphasizing the forest existence of these horses, they received the Latin name Equii silvestris (forest horses). According to historical records, tarpans survived the longest in the wild in the vicinity of Bialowieza Forest, more or less until 1780, when they were finally caught and transferred to the Zoo of the Counts of Zamoyski (an influential Polish count) in the town of Zwierzyniec near the town of Bilhora in Lublin Voivodeship in eastern Poland. Around 1806, the zoo ceased to exist, and the horses were distributed to local peasants. Thus, we can assume that this is the last case of domestication of wild horses in Poland.

The World Wide Fund for Nature (WWF) has launched a program to return the tarpan-like Polish horse to nature. Therefore, tarpan-like horses were brought to the Belarusian part of Belovezhskaya Pushcha and to the south-west of Latvia (1999) [11]. In 2009, with the consent of the Ministry of Environment of Ukraine and the Ministry of Nature Protection of Poland, horses of the descendants of the wild tarpan horse were brought to the territory of Yavoriv National Nature Park from Roztochansky National Park to preserve and reproduce the gene pool of these horses.

However, the origin of the domestic horse and the history of most modern breeds remains unclear to this day. This is due to the fact that the bones of wild and domesticated horses are almost identical. The lack of diagnostic, anatomical and biometric criteria does not allow to determine the archaeozoological remains of the horse.

To solve these problems molecular genetic analysis of horse DNA is used. With the use of molecular markers in research, there are new opportunities to study genetic diversity, as well as the definition of phylogenetic relationships at both intra- and interspecific levels. One of the types of genetic polymorphism research is ISSR-PCR - amplification of intermicrosatellite DNA fragments, the purpose of which is to conduct genetic monitoring in rocks in order to preserve the allelofund of a few rocks. (3)

The aim of the work was to identify intraspecific genetic variability of tarpan-like horses of the Polish konik breed and to establish phylogenetic links between ancient equids (Pleistocene horse, real tarpan) using ISSR - fingerprinting.

The material for the assessment of intraspecific and interspecific genetic variability and the establishment of phylogenetic relationships between modern horses and fossil remains of ancient horses were horses of the Polish konik breed (10 heads) of the Yavoriv National Nature Park of the Lviv Region. and fossil bones of horses of the Pleistocene period (about 10 thousand years BC), metacarpal bone (os. tarsicentral). The bone was found in Novgorod-Siversky, Chernihiv region during quarry construction work. The excavations were carried out by PI Borisovsky in 1935. A tooth found in the village of Tarpan was used to study a wild tarpan horse (4.5 thousand years BC). Skibnytsia, Trostyanets district, Vinnytsia region. Excavations were conducted in 1959 by VM Danylenko. The paleontological material that was studied was provided by the Kyiv National Museum of Natural History of the National Academy of Sciences of Ukraine, Department of Paleontology.

It was found that the species-specific spectra of PCR products obtained by ISSR-PCR for species E.caballus are fragments of 380-400 np and 500-520 np.

The breed-specific fragments of the DNA molecule inherent in horses of the Polish konik breed include spectra of amplification products with a size of 680-710 np.

It was found that the genetic distances between the amplicons of ISSR-PCR markers of Polish horses and fossil remains of a Pleistocene horse were 0.0881 and between the remains of a real tarpan - 0.0845.

The low value of the share of polymorphic loci (P) and the index of polymorphic information content (PIC) was revealed, which indicates a high degree of genetic consolidation of horses of the Polish konik breed and possible reproductive isolation of the population.

It was established that the spectra of amplification products of horses of the Polish horse and fossil remains of Pleistocene horse and real tarpan by 97% on the universal scale of dimension of ISSR fragments in np belonged to "light" (A-35 - A-25) and "medium" -25 - A-13), which is a specific characteristic of E. caballus species.