The Altai falcon from Central Asia always attracted the attention of humans. Long considered a totemic bird in its native area, modern falconers still much appreciated this large-bodied and mighty bird of prey due to its rarity and unique look. The peculiar body characteristics halfway between the saker falcon (Falco cherrug) and the gyrfalcon (F. rusticolus) triggered debates about its contentious taxonomy. The weak phylogenetic signal associated with traditional genetic methods could not resolve this uncertainty. Here, we address the controversial evolutionary origin of Altai falcons by means of a genome-wide approach, Restriction-site Associated DNA sequencing, using sympatric eastern sakers falcons, allopatric western saker falcons and gyrfalcons as outgroup. This approach provided an unprecedented insight into the phylogenetic relationships of the studied populations by delivering 17,095 unlinked SNPs shedding light on the polyphyletic nature of Altai falcons within eastern sakers. Thus we concluded that the former must correspond to a low taxonomic rank, probably an ecotype or form of the latter. Also, we found that eastern sakers are paraphyletic without gyrfalcons, thus, these latter birds are best regarded as the direct sister lineage of the eastern sakers. This evolutionary relationship, corroborated also by re-analyzing the dataset with the inclusion of outgroup samples (F. biarmicus and F. peregrinus), put eastern sakers into a new light as the potential ancestral genetic source of high latitude and altitude adaptation in descendent populations. Finally, conservation genomic values hint at the stable genetic background of the studied saker populations.

Результаты апробирования методики восстановления популяции балобана путём подсадки в естественные гнёзда птенцов, выращенных в питомнике (с результатами GPS/GSM-трекинга слётков)

Mitochondrial DNA variability affects the characteristics of cell metabolism and serves as marker of evolutionary processes. The mitochondrial cytochrome b gene sequences of saker and gyrfalcon were examined. Five amino acid substitutions were identified, and for two of these, changes in amino acid physicochemical properties were revealed. Suggestions on the phylogenetic origin and functional influence of the detected amino acid substitutions were made.

Previous molecular studies have demonstrated significant differentiation of the Strand’s birch mouse (Sicista strandi) into two groups; differences between them are comparable with those observed between species. However, these studies were based on different, not numerous samples and the small number of individuals. To clarify the differentiation pattern and structure of the Strand’s birch mouse, we analyzed variability of the total mitochondrial cytb gene and the nuclear IRBP gene fragment using additional material on this species. Our results confirmed its division into two main genetically distinct forms: the northern form includes Strand’s birch mice from Belgorod and Kursk regions, while the southern form incorporates specimens from the Greater Caucasus as well as Rostov, Saratov, and Lugansk regions. In turn, within the southern form, moderate differences were first revealed by the cytb gene analysis between individuals from Rostov region and samples from the Greater Caucasus and Saratov region. One specimen from Belgorod region had the IRBP haplotypes, typical of this population, but an ‘alien’ mitotype, close to those of Strand’s birch mice from Rostov region. This result may indicate existence of a modern or ancient hybrid zone between the northern form and the ‘Don’ line of the southern form.

DNA analysis is a “gold standard” for individual identification and parentage studies as for humans, so for animals (Saks et al., 1991). First forensic testing of the rare animal species DNA occurred in 1991 in Great Britain, when the case of illegal capture of the Peregrine Falcon (Falco peregrinus) nestlings was proved by parentage testing using DNA fingerprints of birds in question and other peregrines in captivity (Shorrock, 1998). In Russia, illegal capture for foreign sales is the main threat for Saker (Falco cherrug) and Gyrfalcon (Falco rusticolus) populations (Kovács et al., 2014; Lobkov et al., 2020). In 2021, a Comprehensive action plan for creation of Reintroduction and conservation centres for falcons in the Kamchatka region and bustards in the Republic of Kalmykia was signed into law by the Vice-Prime Minister V. Abramchenko. While this statute is primarily addressed to falcon species reproduction and reintroduction, some action items there are coming from falcon species conservation to prevent illegal trade: investigating the orders of marking birds in captivity and tracing them individually from hatching to selling and transferring abroad. Genetic methods were supposed to be the heart of the orders. Accuracy of any DNA identification highly depends on genetic markers used for it. Usually, for humans and farm animals, individual identification and paternity testing are based on nuclear microsatellites (short tandem repeats) panels. These markers are codominant, multiallelic, hypervariable and allow running analyses quickly and automatically. Some studies have already shown their possibilities in forensic DNA testing of falcons (Beasley et al., 2021). We tested different microsatellite panels for genetic identification of Sakers, Gyrfalcons and Peregrines and established a simple and cheap protocol for falcon genotyping using 10 microsatellite loci primarily described for saker natural populations (Hou et al., 2018). Using a first dataset contained of 99 samples (85 Sakers, 12 Gyrfalcons and two hybids), we showed the possibility to estimate the significance of genotyping for these loci and obtained values of probability of identity (PI) less than 10-9 in total and less than 10-3 for siblings. Whereas total population number both for the Saker and Gyrfalcon over the area is estimated by the IUCN in less than 106 individuals, and the offspring of one bird pair can hardly amount to much more than a hundred, the probability of accidental appearance of two birds with same genotypes for these loci may be regarded as negligible. The accuracy of parentage testing was also shown to be sufficient as for the pair and their offspring, so for one parent and the offspring. The microsatellite panel showed specificity for Peregrines, but more investigation is needed to check if these loci are fully convenient also for Peregrine Falcon identification. Basing on these preliminary results we used this test system starting a voluntary programme of breeding stock genotyping for Russian falconries. To the date, five falconries have already provided genetic material and supporting data for their breeding stock, about 380 samples from Sakers, Gyrfalcons, Peregrines and their hybrids. Together with data from natural populations (about 300 samples including museum specimens), this dataset provides opportunities to perform species and population analysis even though our preliminary results like previous data (Nittinger et al., 2007) showed low sensitivity of microsatellites to distinguish Sakers from Gyrfalcons without supporting geographic data. For falconries, full analysis of genetic data, analysis of paternity etc. provided a probability to check the breeding stock and breeding programs and exclude occasional mistakes in identification even for old cases. But the main aim of the program is to obtain a unique genetic identifier for each bird and its offspring to show the legitimacy of rare species keeping. While our microsatellite panel showed genotype uniqueness like CODIS (Budowle et al., 2016) and other forensic test systems for human identification, we propose it as a base of the genetic passport of falcon which can be put into a pedigree sertificate as a plain text or a QR-code. For such purposes, the method should be fully reproducible using any equipment at any laboratory. Together with Russian companies specialized in forensic DNA test systems manufacturing, we plan to develop a ready-to-use kit for genetic passportization of sakers and gyrfalcons in captivity and forensic DNA testing in case these rare species illegal outtake from nature or trafficking is suspected.

Studying intraspecific differentiation in closely related species is essential to clarify the phylogenetic relationships and mechanisms of early stage speciation, particularly in evolutionarily young lineages affected by human-driven population declines. The endangered saker falcon (Falco cherrug), with its ambiguous phylogenetic links to the gyrfalcon (F. rusticolus), exemplifies this scenario. This study presents a comprehensive genetic analysis of F. cherrug and F. rusticolus using mtDNA markers and microsatellite loci, focusing on the diversity of southern Siberian saker falcon populations. The genotyping results for these populations were correlated with phenotypic data obtained from long-term monitoring (1999–2021). Our findings provide novel insights into the current subspecific differentiation and the remnants of a nascent subspecies structure that existed before the recent demographic collapse. Furthermore, our results support the hypothesis of the gyrfalcon’s origin as a descendant species of the Asian saker falcon, i.e., an evolutionarily young lineage undergoing divergence. Our data contribute to the understanding of the Hierofalco evolutionary history, particularly through the analysis of heterogeneous mutation rates among mitochondrial haplogroups. This study underscores the critical importance of conservation efforts for wild endangered populations through long-term monitoring integrated with combined genetic approaches.