Mole small and medium sized insectivores. Body length from about 5 to 21 cm. Tail length from 1.5 to 20 cm. Weight from 9 to 170 gr. Terrestrial running forms, underground and semi-aquatic floating. The body, as a rule, is elongated, smooth, limbs shortened, in most forms adapted for digging, in some for swimming. The brushes are wide, with a bristle of bristled hair and flat claws, palms turned outwards, the feet are narrow and thin. Fingers are armed with long claws flattened from top to bottom. The hind limbs are usually weaker than the front, their feet are narrow and long. The head is small, wide, with an elongated conical muzzle. The eyes are small, in some species they are closed by the skin. The outer auricles are usually absent or very small. The neck is short, sometimes completely invisible. HairlineAs a rule, it is poorly differentiated into categories, thick and soft, one-color, in most cases black, black-gray, black-brown or brown. In aquatic forms, the hair is long and divided into categories. Nipples 3 or 4 pairs.
Skull in a typical case of elongated shape, its greatest width is 2-2.5 times less than its length. Zygomatic arches are available, however very thin. The brain capsule is wide and flattened. There are bone auditory drums.
Teethmoles are small. Dental formula: I 2-3 / 1-3 C 1 / 0-1 P 3-3 / 3-3 M 3/3 = 34-44. Incisors, fangs and pre-radicals are highly variable in shape, located on the front edge of the upper and lower jaws. The vertebrae of the cervical 7, thoracic 13-14, lumbar 5-6, sacral 5-6, caudal 8-27. Two sacral and three first caudal vertebrae fuse together, their spinous processes form a highly developed crest. The collarbone is very short and wide. The sternum handle is long, with a high longitudinal crest and short lateral processes. The shoulder blade of the most adapted to digging is long, its greatest width is 5-6 times less than the length, and the humerus is short and wide throughout. The humerus in moles is extremely wide and shortened. The collarbone articulates directly with the humerus - a case unique to mammals. The pelvic bones are long and narrow. The pelvis is still connected to the spine. The pubic bones do not form a ventral symphysis. Males have os penis. The cecum is absent.
The inhabitants different landscapes. Most species lead an underground lifestyle. They dig tunnels at various depths and, as a rule, feed on underground. Some species in search of food periodically come to the surface of the soil. A number of species lead a terrestrial lifestyle, although burrows are a refuge for them. Finally, there are water forms that swim and dive beautifully, make burrows on the banks of water bodies. Activity round-the-clock or twilight-night. They usually stay alone.
Pregnancy about 5 weeks. The female appears to bring offspring once a year. Maturity occurs at the age of about a year. Eat various invertebrates: terrestrial, underground and aquatic. Some species are subject to fishing. Eating a large number of harmful invertebrates can be of substantial benefit to agriculture and forestry. They do some harm by eating earthworms.
Common in North America, from southern Canada to northern Mexico, in Europe and Asia, south to Malaya and the Mediterranean Sea, and north to about 63 ° C. w. The taxonomy of the mole family has some contentious issues.
Main stream family specializations well expressed in species adapted for digging underground. Their morphological features reflect the shape of the body and limbs that is most advantageous for digging and moving in narrow burrow passages. This type of movement is most common within the family and is characteristic of most species. A smaller number of species leads a land-based lifestyle, and their adaptations to digging have the character of the initial stages of the formation of these limb functions. Many of them run relatively quickly on the surface of the soil. The structure of the chewing apparatus of moles reflects the adaptation to feeding mainly small invertebrate animals, soft consistency.
There are 2 kinds in the CIS: desman and moles, common in the forest and forest-steppe zones of the European part of the USSR, Western and Central Siberia to southeastern Transbaikalia, in the Primorsky Territory.
Most species of moles are subject to fishing. Useful value they also consists in the extermination of harmful animals. Harmful activity expressed in soil degradation as a result of eating earthworms, contributing to loosening the soil layer, and in throwing the earth from deep layers to the soil surface. The latter leads to the formation of bumps, the deterioration of meadows and pastures. In addition, digging, moles damage the root system of plants.
Late Eocene - modernity, Europe, Asia, Oligocene - modernity, North America. It includes three subfamilies and seven tribes, two of which are extinct. There are two subfamilies in the USSR: Desmaninae Thomas, 1912 (Late Eocene - present) and Talpinae Fischer v. Waldheim, 1817 (Late Eocene - present). Probably finding fossil remains of representatives of the subfamily Urosilinae Thomas, 1912.
1. Sokolov Century E. Systematics of mammals. Textbook manual for universities. M., "Higher School", 1973. 432 pp. With ill.
2. Catalog of mammals of the USSR (Pliocene-modernity). L., "Science", 1981. 456 p.
3. Mammals of the USSR. Part 1. Publishing House of the USSR Academy of Sciences. Moscow-Leningrad, 1963
4. Naumov N. P., Kartashev N. N. Zoology of vertebrates. - Part 2. - Reptiles, birds, mammals: Textbook for biologist. specialist. un-com. - M .: Higher. School, 1979.- 272 s, ill.
- Mammals Big Encyclopedic Dictionary / Ed. AND I. Pavlinova. M .: ed. AST, 1999.416 s.
- Mammals of the USSR. T. 1. / Ed. I.I.Sokolova. L .: Nauka, 1963.638 p.
- Sokolov V.E., Tembotov A.K. Vertebrates of the Caucasus. Mammals: insectivores. M .: Nauka, 1989.554 s.
General characteristics of the tribe Talpini
The family of moles (Talpidae), together with hedgehogs, shrews, and alkalines, is part of the true insectivorous order (Eulipotyphla) (Murphy et al., 2001, Douady, Douzery, 2003).
In modern surveys on mole taxonomy, there are from 12 to 17 genera (Gureev, 1979, Corbet, Hill, 1992, Motokawa, 2004, Hutterer, 2005), the relationships of which are contradictory - and this applies both to systems based on morphological features of the skull and teeth ( Motokawa, 2004), osteological characteristics (Sanchez-Villagra et al., 2006), and the results of comparison of mitochondrial DNA sequences (Shinohara et al., 2003, 2004b, 2014, He et al., 2014). Thomas (Thomas, 1912, cited by Nikolsky, 2002) and Cabrera (Cabrera, 1925, cited by Nikolsky, 2002) distinguished five Talpidae subfamilies based on the structure of the skull and teeth: Uropsilinae, Desmaninae, Condylurinae, Scalopinae, Talpinae. Nikolsky (2002) adheres to Campbell's systematics (Campbell, 1939), in which the Talpidae family is divided into six subfamilies: Uropsilinae (shrew moles), Desmaninae (muskrats), Condylurinae (starfish), Urotrichinae (lower true moles), Scalopina North America), Talpinae (the highest real moles of Eurasia). According to the latest edition of Mammal Species of The World (Wilson and Reeder, 2005, hereinafter MSW3), the family includes 3 subfamilies: Talpinae (underground moles), Scalopinae (the highest true moles of North America) and Uropsilinae (shrew moles). The Talpinae subfamily, which includes species belonging to five tribes, is the most numerous in the number of genera and species: Desmanini, Neurotrichini, Scaptonychini, Urotrichini, and Talpini (Hutterer, 2005).
The tribe Talpini was first isolated by Fisher in 1817 (Fischer von Waldheim, 1817, cited by Gureev, 1979). Representations of the phylogeny and systematics of the Talpini tribe were based mainly on the morphology of the skull, teeth, and postcranial skeleton. According to morphological and odontological characteristics, Stroganov (1948) distinguished 6 genera in its composition: Talpa, Mogera, Scaptochirus, Euroscaptor, Eoscalops and Parascaptor. Schwarz (1948), based on the dental formula and believing that these are only variations of the same genus, brought the genera Asioscalops, Mogera, Scaptochirus, Euroscaptor, Eoscalops, and Parascaptor into synonyms for the genus Talpa. Stein (1960), using the allometric method for studying skull structures, singled out such genera as: Mogera, Talpa, Parascaptor, Scaptochiru s. According to Hutterer (2005), the Talpini tribe includes five genera: Euroscaptor, Mogera, Talpa, Parascaptor, Scaptochirus. In this case, the genus Talpa is distributed from Western Europe to Western Siberia, while the remaining four genera are in East and Southeast Asia.
The relationship between genera according to morphological data is debatable. Motokawa (Motokawa, 2004), based on the morphology of skulls, showed that representatives of the genus Talpa occupy a basal position in relation to the other genera of the tribe Talpini, in the same work paraphilia of the genus Euroscaptor was revealed. However, an analysis of osteological data (Sanchez-Villagra et al., 2006) revealed the basal position of the genus Parascaptor and sister relationships of the genera Mogera and Scaptochirus, later on the molecular data of Japanese and Chinese researchers (Shinohara et al., 2008, He et al., 2014) , as well as the results of our work (Zemlemerova et al., 2013), which are presented extensively in the “Results” and “Discussion” sections, confirmed Motokawa's morphological point of view (Motokawa, 2004) on the basal position of the Talpa genus and on the paraphilia of the Euroscaptor genus.
History of the study of the genus of common moles Talpa The central genus of the tribe Talpini is the most widespread genus Talpa. Its range covers the entire Palearctic region, from the Iberian Peninsula to Siberia. Some authors include Asian scaptor Miller, 1940 (Corbet, 1978, Gureev, 1979, Corbet, Hill, 1992) and Mogera (Corbet, 1978) as Asian subgenus. The number of species recognized in the Talpa genus has changed over time as new characters and research methods have been attracted. Many modern species were originally described as subspecies of T. europaea, T. romana, T. caeca (Filippucci et al., 1987).
According to qualitative morphological and morphometric characteristics (eyelids, open eyes, body length, M2 width, I1 height, claw width of the third finger of the hand, etc.), Stroganov (1948) identified six species in the genus Talpa: T. europaea, T .caucasica, T. orientalis (currently this species belongs to the subspecies T. caucasica), T. romana (as a subspecies of which Stroganov considered T. stankovici, which is currently isolated as a separate species), T. caeca (as a subspecies of which he considered T. occidentalis and T. levantis, which are currently isolated in separate species) and T. altaica.
Gureev (1979) distinguished 12 species: T. minuta, T. europaea, T. altaica, T. caeca, T. caucasica, T. romana, T. streeti (= Talpa davidiana), T. k lossi, T. grandis, T. parv idens, T. longirostris, T. micrura. Currently, the last five species belong to the genus Euro scaptor.
Currently, taking into account morphological, karyological, and molecular data, nine species of the genus Talpa are distinguished: four are distributed from the Middle East to Siberia and Altai: T. altaica Nikolasky, 1883, T. levantis Thomas, 1906, T. caucasica Satunin, 1908, T. davidiana Milne-Edwards, 1884, four from Western Europe: T. caeca Savi, 1882, T. romana Thomas, 1902, T. occidentalis Cabrera, 1907, T. stankov ici Martino and Martino, 1931 and distributed throughout Europe T europaea Linnaeus, 1758 (Pavlinov, 2003, Hutterer, 2005, Pavlinov, Lisovsky, 2012).
Amplification and sequencing of individual genes
Talpini is one of the most monotonous, both in appearance and in the nature of adaptations, a group of moles (Gureev, 1979). Unlike the semi-underground and terrestrial moles from the tribes Urotrichini, Neurotrichini, Scaptonychini, representatives of the tribe Talpini have a number of morphological features in connection with the underground lifestyle: the head is not separated from the body by neck interception, the auricles are not developed, the eyes are rudimentary or often completely covered by skin ( Gureev, 1979). The development of the locomotor system went in the direction of increasing digging efficiency, which made it possible to tear tunnels not only in light but also in heavy soil. The appearance of the clavicular-shoulder joint and subsequent transformations of the locomotor apparatus occurred. There was a direct connection between the shoulder and the axial skeleton (in addition to the scapula), which makes it possible to practically unlimitedly increase the lateral forces applied to the substrate (Nikolsky, 1978). In addition, there was hypertrophy of the muscles of the forelimb, expansion of the humerus and hand, shortening of the clavicle, lengthening of the sternum, shortening of the lower leg and foot (Nikolsky, 2002).
An interesting feature was found in the structure of the reproductive system in T. romana, T. europaea, T. occidentalis, and T. stankov ici, probably related to the peculiarities of the late lifestyle and behavior. Females of these species have the so-called ovotesticules - gonads containing tissues and eggs and testicles (Jimnez et al., 1993, Sanchez et al., 1996, Whitworth et al., 1999).
In this case, the females remain fertile, while the testes cells are sterile and function only for the production of testosterones. Among other representatives of the Talpidae family, this feature was found in M. wogura (Carmona et al., 2008), Galemys pyrenaicus (Peyre, 1961, cited by Carmona et al., 2008), Condilu racristata (Rubenstein et al., 2003 ), however, in Urotrichus talpoides (Carmona et al., 2008), Scalopus aquaticus, Scapanulus latimanus, and Scapanulus orariu s (Rubenstein et al., 2003), such a feature was not found. Beolchini et al. (Beolchini et al., 2000) suggest that there is a relationship between the presence of female androgens in the body and the absence of pronounced sexual behavioral demorphism in moles. In addition, the sensitivity of muscle tissue to the presence of androgens in the body of both males and females has been previously shown (Krieg, 1976, cited by Beolchini et al., 2000) and, most likely, this is due to the fact that females, like As males, strong musculature is required to optimize digging processes (Beolchini et al., 2000). However, Rubenstein et al. (Rubenstein et al., 2003) refuted this assumption, as muscular development was also observed in females with normal ovaries. Jimenets et al. (Jimnez et al., 1993) showed seasonal dependence of testosterone levels in both males and females, while females showed low hormone levels during the breeding season, while males showed the opposite situation. Wingfield (2005) suggested a relationship between testosterone levels and aggressiveness and territoriality. It is likely that seasonal changes in testosterone levels cause changes in the behavior of females during the breeding season (Carmona, 2006, cited by Carmona et al., 2008). To confirm this hypothesis, data on the level of the hormone in North American species are required. The fact that moles are highly territorial has been repeatedly confirmed by different authors (Ognev, 1928, Stroganov, 1948), and the boundaries of the territories of individual individuals do not overlap, overlapping areas are observed only between animals of different sexes (Loy et al., 1992). Ognev (1928) wrote: “this predatory, non-living animal, according to most authors, lives a hermit in his hunting possessions, hostile to his brother or any living creature that he encounters in an underground gallery. If he comes across a shrew, a mouse, a toad, or even so, the mole mercilessly kills and eats its victim. They say that two moles, especially if they are of the same sex, engage in a mortal struggle: the winner kills the vanquished and eats him. ” However, a direct relationship between the structural features of the female mole reproductive system and other morphological and molecular characters was not found (Carmon et al., 2008).
The development of moles associated with digging occurred in the direction of the development of a certain food resource - earthworms. The creation of moving systems in the thickness of the soil, sometimes at great depths, and not just moving in the plant litter, provided moles with a number of advantages. Their protection from predators and adverse abiotic conditions increased, and the severity of competitive relations decreased. The created moves still serve as peculiar traps for many invertebrates, and for immobilized earthworms as a place of long storage (Gureev, 1979). A common ecological feature of moles is their attachment to tree-shrub vegetation and the territories located close to them (Gureev, 1979). One of the important factors affecting the distribution of moles is the type of soil, its physical and chemical parameters. Moles prefer fertile soil: meadows, pastures and agricultural land, as well as gardens, deciduous and mixed forests. In coniferous forests are quite rare. Avoid sandy, stony soils where digging is difficult or impossible. In addition, the degree of soil moisture is important for moles. They choose a moist but not swampy environment. They are found in periodically flooded areas, but are absent in areas with a constant high level of groundwater (for a review see Zurawska-Seta, 2010,
urawska-Seta, Barczak, 2012). An essential factor for moles is the availability and accessibility of food. Mostly moles feed on Lumbricidae earthworms and insect larvae.Also, their diet may include pupae and adult insects, spiders, millipedes, snails, sometimes larger animals such as frogs, lizards, snakes, mice and other small rodents, and even representatives of their species (for a review see Beolchini, Loy, 2004, urawska-Seta, 2010). The ratio between earthworms and other invertebrates in the diet of the mole is quite constant and depends on the time of year and the environment. An important factor is the pH of the soil. Earthworms are numerous in soils with a pH above 4.0, and the optimal conditions for moles are soils with a pH close to neutral in value, so moles avoid acidic soils. A link has also been established between the distribution of moles and grazing sites for pets (for a review see urawska-Seta, 2010 and urawska-Seta, Barczak, 2012).
Summing up all the above, we can say that the peculiar way of life and the occupied ecological niche could not but affect the morphology and diversity of moles.
As was shown, modern ideas about the phylogeny and systematics of the Talpini tribe are based on the morphology of the skull, teeth, and postcranial skeleton. Attraction to the systematics of moles of sequencing data began relatively recently. The difficulty of determining, the subjectivity in assessing the significance of various morphological diagnostic characters and the instability of the main diagnostic character - teeth - have determined significant differences in the views on the volume, composition and relationships of mole taxa not only of species but also of genus.
The dental system (the structure of incisors and molars) is the basis of modern diagnosis of mole species. The very first classifications were based solely on odontological and craniological features. Not only Talpa moles, but also Asian births are classified mainly by the dental formula. The first mole taxonomy based on this trait was made by Gill (Gill, 1875). Subsequently, Dobson (Dobson, 1883) and Allen (Allen, 1938) followed this idea:
Phylogenetic analysis of individual nDNA genes
The sequences of the mitochondrial cytb gene (cytochrome b) and COI (I subunit of cytochrome oxidase) and fragments of five exons were analyzed: RAG1 - recombination activating gene 1), BRCA1 - breast cancer gene 1 (exon 11 of the breast cancer type 1 susceptibility protein), BRCA2 - gene for breast cancer 2 (exon 11 of the breast cancer type 2 susceptibility protein), ApoB - gene for apoliprotein B (apolipoprotein B), A2ab - gene for -2 adrenergic receptor (-2 b adrenergic receptor).
The polymerase chain amplification reaction (PCR) for six genes was carried out in a volume of 20 μl. For the reaction, a mixture was prepared for the required number of samples, consisting of two primers, Taq-buff, 25 mM MgCl2, dNTP 2.5 mM, Taq polymerase and water. Sileks Taq polymerase (5 units / μl) was used at a concentration of 0.2 μl per sample.
The primer sequences are shown in table 9. The complete sequence of the mitochondrial cytochrome b gene (1140 bp) was amplified with combinations of primers L14734 / H15906, L_talpa_glu / H_talpa_pro. To read the internal sections, additional primers L393 and H525 were used. A portion of the RAG1 nuclear gene sequence (1010 bp) was amplified with combinations of primers F1851 / R2951, and primers F2401 / R2486, F319 / R745, R320 / F730 were used to read the internal regions. A portion of the BRCA1 nuclear gene sequence (1263 bp) was amplified with combinations of primers F120 / R1020 / R1330; additional primers F437 / R700 were used to read the inner sections. Plots of the nuclear gene sequences BRCA2 (1113 bp), ApoB (975 bp) and A2ab (852 bp) were amplified with various combinations of the primers presented below.
DNA extracted from six museum specimens of T. caucasica ognev i, eight specimens of T. levantis minima, and also two specimens of M. robusta was severely degraded; therefore, for such specimens of the genus Talpa, only short cytb fragments were obtained using a combination of internal primers L393 / H525. To ensure the reliability of the resulting product, four additional internal primers were developed for amplification of fragments 260-400 bp long using a combination of primers: L148 / H510, L148 / H330a and L50 / H330a. For amplification of museum samples of M. wogura, primer combinations were used: L48 / R270A, F97 / R176 and F172 / R270A.
Amplification was performed on My Cycler (Bio-Rad, USA) and Mastercycler Gradient (Eppendorf, Germany) devices. Amplification was monitored on a 1% agarose gel. Purification of PCR products for sequencing was carried out by precipitation with a mixture of ammonium acetate with 70% ethanol or a ready-made set of reagents "Diatom DNA Elution" (OOO Isogen Laboratory).
The bulk of the experimental work was performed in the office of "Molecular Methods in Zoology" of the Department of Vertebrate Zoology of Moscow State University. M.V. Lomonosov.
Automatic sequencing was performed on an ABI 3100-Avant sequencer using the ABI PRISM BigDyeTM Terminator v. 3.1 in the laboratory of the CCU GENOM.
Sequence alignment was performed manually using DNAStar Lasergene SeqMan Pro v. 7.1.0 (Burland, 1999) and BioEdit v. 126.96.36.199 (Hall, 1999).
To partition the concatenated nucleic acid sequence into partitions, the PartitionFinder v1.0.1 program was used (Lanfear et al., 2012). Initially, five theoretically possible schemes were proposed: (1) a breakdown by genes, (2) a breakdown by codon positions, (3) a breakdown by genes and codon positions (three subgroups for a gene), (4) as in option 3, but 1- I and the 2nd position of the codons are combined (two subgroups for the gene), (5) without splitting. The best option for the Talpa and Euroscaptor genera and for determining the position of the genera in the Talpini tribe was the fourth type of partition. For the genus Mogera, the second type of partition was optimal. For cytb data, a codon splitting has always been used.
It is known that nucleotide bases in the 1st and especially in the 3rd positions are most subjected to multiple direct and reverse substitutions (Hassanin et al., 1998), which leads to saturation of the sequence and distortion of the phylogenetic signal (Bannikova, 2004). This is especially true for rapidly evolving sequences; in our study, such a gene is cytb. Therefore, in the analysis of the phylogenetic relationship of genera in the Talpini tribe for the cytb gene for the 1st and 2nd positions, all substitutions were taken into account, but for the 3rd position only transversions were taken into account, and when analyzing the concatenated sequence of 5 nuclear genes and cytb for nuclear genes, all substitutions at all positions of the codons were taken into account, and for cytb, transversions of the 1st and all substitutions of the 2nd position of the codons were taken into account.
Reconstruction of phylogenetic trees Reconstruction of phylogenetic trees was performed using several widely used algorithms: maximum parsimony (maximum parsimony, MP), maximum likelihood (maximum likelihood, ML) and Bayesian analysis (MrBayes, BI). Phylogenetic analysis using the maximum economy (MP) method was performed in the PAUP program, version 4.0b10 (Swofford, 2003). The construction of trees by the maximum likelihood (ML) method was performed using the program package Treefinder, version October 2008 (Jobb, 2008). Reconstructions of ML trees were preceded by determining the optimal model of sequence evolution using the PartitionFinder v.1.0.1 program (Lanfear et al., 2012) based on the BIC criterion (Bayesian information criteria). The optimal models for each genus and gene are presented in table 10.
To check the stability of the treasure, a bootstrap procedure with 1000 pseudo-replies for MP, NJ and ML algorithms was used.
Bayesian analysis (BI) was performed in MrBayes 3.1.2 software (Ronquist, Huelsenbeck, 2003). For this analysis, we used the MCMC algorithm and the following parameters: the number of independent analyzes 2, the number of generations of 20 million each, the 5,000th generation, the number of Markov chains were selected in the pseudo-sample 8. The convergence was estimated using the ESS statistics (effective sample size) in Tracer 1.4 (Rambaut, Drummond, 2007).
The species tree was built in the BEAST program (Heled, Drummond, 2010). The following set of a priori assumptions and parameters was used: the relative rate of formation of new lines was considered constant (Yule prior), nucleotide evolution models were used for different positions of the codon in accordance with Table 10. The division into groups was carried out in accordance with the division obtained using ABGD ( Automatic Barcode Gap Discovery method) (Puillandre et al., 2012) (see parameters below). The molecular evolution rate was analyzed in PAML 4.7 (Yang, 2007). According to the data obtained, for the genus Talpa, strict clock models were used for nuclear genes, and for non-strict clock models for cytb, while the distribution of velocities along the tree branches corresponds to the uncorrelated lognormal relaxed clock model, for the genus Euroscaptor - a model of non-strict watches was used for the cytb and BRCA1 genes, for the remaining nuclear genes - strict watches. The average rate of evolution was considered equal to unity, the number of independent analyzes was four, the chain length was 20 million generations, and every 5000th generation was selected in a pseudo-sample. Convergence was assessed using ESS statistics (effective sample size) in Tracer 1.4. (Rambaut, Drummond, 2007).
Comparison of the results of phylogenetic analysis of the mitochondrial cytb gene, the combined sequence of nuclear genes and the species tree
The final cytb alignment for 26 samples, including 5 external groups, was 1140 bp. Application of the PartitionFinder (Lanfear et al., 2012) to a set of five nuclear genes showed that the best scheme according to the BIC criterion corresponds to 10 partitions (partitions by genes and codon positions when combining the 1st and 2nd positions). The best models for each of the 10 partitions of nuclear genes and the three cytb positions are shown in Table 10 (section “Material and Methods”).
In a combined analysis of the mitochondrial cytb gene and five nuclear genes, the final alignment consisted of 5812 nucleotide positions, including 760 bp cytb, 960 bp ApoB, 1245 bp BRCA1, 807 bp A2ab, 1032 bp BRCA2 and 1008 bp RAG1. The total data set included 15 sequences, including 4 from the external group. Phylogenetic analysis of the mitochondrial cytb gene The principle topology of the trees obtained on the basis of the phylogenetic algorithms ML and BI is the same. In fig. 25 presents an ML tree with significant bootstrap support in BI / ML analyzes.
In the presented tree, the Talpini tribe shows a clear monophilia of groups corresponding to the genera Talpa and Mogera, while the genus Euroscaptor is paraphyletic due to the position of Scaptochirus and Parascaptor inside its radiation. The entire group, including Asian mole births (Mogera, Euroscaptor, Scaptochirus and Parascaptor), is monophyletic. Fig. 25. Results of ML analysis of 1140 bp cytb. The numbers near the tree nodes are indicators of bootstrap supports in BI / ML analyzes, respectively. An asterisk indicates absolute support in both analyzes. Combined analysis of nuclear genes and mitochondrial cytb gene
In fig. Figure 26 presents an ML tree constructed from a combined analysis of the total sequence of nuclear genes (ApoB + BRCA1 + BRCA2 + RAG1 + A2ab) and the mitochondrial cyt b gene with significant bootstrap supports in BI / ML analyzes. The results reproduce the monophilia of the genera Talpa and Mogera revealed in the mitochondrial analysis, as well as the group consisting of Asian species (Mogera + Euroscaptor + Parascaptor + Scaptochirus). The genus Euroscaptor is still paraphyletic due to the isolated position of E. mizura outside the cluster of other species of the genus.
ML tree of the combined gene sequence ApoB + BRCA1 + BRCA2 + RAG1 + A2ab + cytb. The numbers near the tree nodes are indicators of bootstrap supports in BI / ML analyzes, respectively. An asterisk indicates absolute support in both analyzes. Consider the relationship of species in each genus separately. Phylogenetic analysis of the mitochondrial cytb gene
In the analysis of the mitochondrial cytb gene (1140 bp), an extended sample of the Talpa genus was used compared to nuclear genes - 114 samples and 27 for the external group.
The obtained ML-tree with high supports confirms monophilia of almost all species of the Talpa genus, with the exception of two cases: (1) T. europaea, which turned out to be paraphyletic relative to T. occidentalis, (2) T. levantis, whose Talysh population, known as a subspecies T. levantis talyschensis Vereschagin, 1945, was far removed from the rest of the small mole. In this regard, in the rest of the work we will denote this grouping as Talpa sp.
T. europaea is paraphyletic due to the isolated position of seven samples from the four localities of Northern Spain - Haro, Fogas de Monclus, Montseny, Sant Pere de Vilamor. K2P distances between the Spanish group and other groups of T. europaea are 8% (Table 11). The high genetic distances that separate it from the others, the group and the peripheral geographical location indicate that the taxonomic status of this North Spanish population requires further close study. With high probability, it does not belong to the species T. europaea (Feuda et al., 2015).
The remaining samples of T. europaea comprise a monophyletic sister group with T. occidentalis. Another supported sister group is T. caeca + T. romana. Treasures T. europaea / T. occidentalis and T. caeca / T. romana in turn are sisterly. The position of T. stankov ici and T. levantis is not permitted.
T. altaica occupies a basal position, but bootstrap support is low (0.96 / 64 / in BI, ML, and MP analyzes, respectively), and the AU test rejects the hypothesis of the position of T. altaica inside genus radiation (p = 0.033).
Genetic distances (K2P) between species vary from 9.1 to 15.6% (Table 11): the minimum distance is observed between T. europaea and T. occidentalis (9.1 ± 0.9) and between T. stankov ici and T. levantis ( 9.1 ± 0.8), the maximum is between T. davidiana and T. altaica (17.0 ± 1.4). The DNA of T. davidiana was analyzed for the first time. T. dav idiana forms a monophyletic group with Talpa sp. from Talysh, the genetic distance between them is 13.2%.
Within most species, deep intraspecific groups can be distinguished that correspond to certain geographic localities. Genetic distances (Table 12) between which vary from 2% between the Italian and European groups of T. europaea to 10% between the groups of T. caucasica. Inside T. caucasica are samples from the North Caucasus and northeastern Turkey. Inside T. levantis is a group that includes specimens from the North Caucasus, Transcaucasia, northeastern Turkey and northern Turkey. T. caeca is divided into Italian and Balkan groups, T. romana - into groups from central and southern Italy, T. occidentalis - into a group from Portugal and southeastern Spain. Three groups are distinguished within T. stankovici: Peloponnese, Macedonia + western and northwestern Greece, and a group from central Greece. Within T. europaea (excluding the Spanish line), two intraspecific groups can be distinguished: Italian and Central-West European, although they have moderate bootstrap support. Within T. altaica with moderate bootstrap supports, groups are distinguished from the Yenisei coast (Mirnoye village) and from the coast of Lake. Teletskoye.
In addition to samples from Turkey and the Caucasus, we analyzed Caucasian moles from Georgia - T. caucasica ognevi, which were not included in the phylogenetic analysis due to the short length of the sequenced nucleotide sequences. For T. c. ognevi received a length of only 380 bp The resulting fragments T. c. ognevi were very similar (1.5%) to the corresponding fragments for the complete nucleotide sequence of samples from Turkey, including all diagnostic substitutions that distinguish populations of moles of the North Caucasus from moles of Turkey (Fig. 28 A).
The genetic distance between T. levantis minima from the village of Nikitino and samples from Kabardino-Balkaria, Armenia or northeastern Turkey (Ardahan) is 2.5%. In addition, we analyzed museum samples of T. levantis minima from Adygea, which were also not included in the phylogenetic analysis due to the short length of the sequenced nucleotide sequences — 253 bp. The genetic distance between the sequences of samples from Adygea and the corresponding sections of the sequences of small moles from Kabardino-Balkaria, Armenia, or northeastern Turkey (Ardahan) (Fig. 28 B) is 3%.