[Microsatellite loci variation and investigation of gene flow between two karyoforms of Cricetulus barabensis sensu lato (Rodentia, Cricetidae)].

We examine the diversity of six microsatellite loci and partial RAG1 exon of "barabensis" and "pseudogriseus" karyoforms in Cricetulus barabensis sensu lato species complex. A total of 435 specimens from 68 localities ranging from Altai to the Far East are investigated. The results of the population structure analysis (factor analysis and NJ tree based on Nei genetic distances) support subdivision into two well-differentiated clusters corresponding to the two karyoforms. These karyoforms are also well differentiated by the level of microsatellite variability. In several "barabensis" specimens, we found microsatellite alleles that are common in "pseudogriseus" populations but are otherwise absent in "barabensis." Most of these specimens originate from a single population in one of the zones of potential contact between karyoforms, Kharkhorin in Central Mongolia. These molecular results are consistent with previously published karyological data in suggesting that rare hybridization events between the two chromosomal races occur in nature.

[DNA CONTENT IN INDIVIDUAL CHROMOSOMES OF NORMAL CHINESE HAMSTER KARYOTYPE AND CHROMOSOMES OF CONSTANT HAMSTER CELL LINES CHL V-79 RJK AND Vebr-5].

Using cytometry and an microfluorimetry, we have determined the genome size in Chinese hamster Cricetulus griseus, as well as absolute and relative DNA content of its individual chromosomes and of chromosomes in the transformed Chinese hamster cell lines V-79 RJK and Vebr-5 after prolonged cultivation. It has been shown that the genome size in male and female Chinese hamster is 6.660 and 6.746 pg, respectively. Absolute content of chromosomal DNA of both studied cell lines differed significantly from the content of the corresponding chromosomal DNA of the Chinese hamster normal karyotype. During long-term cellular cultivation, changes in the DNA content of certain chromosomes in both cell lines (generally upward) reached 20-25 %. The level of DNA amplification in the p-arm of chromosome Z6, registered at the beginning of the experiment, in the course of further cellular cultivation (over 20 years) remained stable. The data obtained allow us to conclude that the malignant transformation of cells and subsequent adaptation to the conditions in vitro leads to a profound restructuring of its genome, which affects almost all chromosomes.

[Genetic diversity and phylogenetic relationships of the Manchurian zokor Myospalax psilurus (Rodentia, Muridae) according to RAPD-PCR analysis].

The population structuring and low genetic diversity of the Manchurian zokor Myospalax psilurus Milne-Edwards, 1874, an East Asian endemic included in the Red List of Russia, were demonstrated. Two separate geographical groups differing in the level of their genetic diversity were found on the territory of the Primorskii krai. The subpopulation located closest to the main area of this species was determined as ancestral. A subspecies differentiation of the Primorskii krai and Transbaikal M. psilurus populations was shown, as was the monophyletic origin of M. psilurus and its high divergence from M. aspalax. The animals from northern localities are recommended for reintroduction in nature under species recovery programs in Primorskii krai.

[Genetic diversity and phylogenetic relationships in the zokor subfamily Myospalacinae (Rodentia, Muridae) inferred from RAPD-PCR].

Zokors (Myospalacinae) is a group of rodents specialized for underground life, endemics of eastern Asia, which is taxonomically and evolutionarily poorly studied. We examined genetic diversity and phylogenetic relationships among zokors (Myospalax myospalax, Myospalax aspalax, Myospalax armandii, Myospalax psilurus, Myospalax smithii) using RAPD-PCR. The subfamily Myospalacinae was shown to be monophyletic and contain four evolutionary branches: M. myospalax, M. aspalax-M. armandii, M. smithii and M. psilurus. Genetic differences and high differentiation were found among the species and between two geographic forms of Manjurian zokor M. psilurus from the marginal parts of the range, Transbaikalia and Primorye. The psiluris phylogroup was shown to be dichotomically divided into two clades according to the geographical distribution of animals from Transbaikalia and Primorye. The genetic differentiation between the geographic forms of M. psilurus corresponded to the differentiation between morphologically similar species M. aspalax and M. armandii. M. armandii is a sister taxon with regard to M. aspalax. This new evidence on the evolutionary relationships among zokors does not contradict the traditional views inferred from morphological, karyological, and allozyme data, on isolation of M. myospalax and the character of form development in this group. The species status of Myospalax psilurus Milne-Edwards, 1874, M. epsilanus Thomas, 1912, M. armandii Milne-Edwards, 1867, which had been suggested earlier on the basis of biochemical and karyological data, was confirmed.

[Molecular genetic relationships among east Palearctic ground squirrels of the genus Spermophilus (Sciuridae, Rodentia)].

Genetic diversity in the four east Palearctic ground squirrel species of the genus Spermophilus--S. undulatus, S. parryi (subgenus Urocitellus), S. dauricus, and S. relictus (subgenus Citellus)--- was investigated using RAPD PCR with ten random primers. Siberian chipmunk, Tamias sibiricus, was used as an outgroup. Molecular markers for different taxonomic ranks were identified, including those for the genera Spermophilus and Tamias, subgenera Urocitellus and Citellus, as well as for each of the four species, S. undulatus, S. parryi, S. dauricus, and S. relictus. For the ground squirrel species and subgenera, genetic differentiation indices (H(t), H(s), D(st), G(st), Nm, and D) were calculated. In addition, for these groups the NJ phylogenetic reconstructions and UPGMA dendrograms of genetic similarity of the individuals and combined populations were constructed. Comparative molecular genetic analysis revealed a high genetic differentiation between S. undulates, S. dauricus, S. relicts, and S. parryi (G(st) = 0.58 to 0.82; D = 0.53 to 1.06), along with a low level of genetic differentiation of the subgenera Citellus and Urocitellus (G(st) = 0.33; D = 0.27), distinguished in accordance with the existing taxonomic systems of the genus Spermophilus.

[RAPD-PCR analysis of ground squirrels from the Tobol-Ishim interfluve: evidence for interspecific hybridization between ground squirrel species Spermophilus major and S. erythrogenys].

Populations of two ground squirrel species, Spermophilus major and S. erythrogenys, from the interfluvial area of the Tobol and Ishim rivers, where their ranges overlap, have been examined using RAPD-PCR. We have identified 253 loci, which included taxon-specific markers for S. major and S. erythrogenys as well as markers for geographic populations. Estimation of genetic diversity and construction of phylogenetic relationships were performed using software programs POPGENE, TEPGA, and TREECON. In all, based on morphological traits, animals from the Tobol-Ishim interfluve were assigned to the two parental morphotypes and showed similar levels of genetic variability (H, n(a), n(e)). However, the total polymorphism level proved to be higher in ground squirrels with the major morphotype (P = 40.32%, P95 = 27.27%) than in animals with the erythrogenys morphotype (P = 32%, P95 = 22.13%). Nevertheless, the number of rare alleles was high in both cases, constituting about 70% of the total number. By contrast, interpopulation differentiation was considerably higher in S. major (6 = 0.50) than in S. erythrogenys (delta = 0.41). The genetic differentiation between local samples from the Tobol-Ishim interfluvial area was lower than that between the parental species. A significant part of the genetic diversity of the species examined and animals from the zone of overlapping ranges was accounted for by intrapopulation variability. Animals from the northern and southern parts of the Tobol-Ishim interfluve were characterized by the core traits of S. major and S. erythrogenys, respectively, falling into two distinct clusters in the UPGMA and NJ reconstructions. In addition to three hybrid individuals, identified by the bioacoustic method, three hybrid animals were distinguished using RAPD analysis. These animals earlier were thought to be "pure" species and formed their own clusters in phylogenetic reconstructions. Thus, the RAPD-PCR results directly showed the existence of stable hybridization (20% genetic hybrids) between S. major and S. erythrogenys in the Tobol-Ishim interfluvial area, which is more extensive than inferred previously from morphological and bioacoustic data.

[Allozyme differentiation of various chromosomal forms of the spotted suslik (Spermophilus suslicus Guld, 1770, Rodentia)]. / Allozimnaia differenttsiatsiia raznokhromosomnykh form krapchatogo suslika (Spermophilus suslicus Guld, 1770, Rodentia).

Eleven enzyme and four nonenzyme protein systems controlled by 25 loci were electrophoretically analyzed in the allopatric karyotypic forms (2n = 34 and 2n = 36) of the spotted souslik Spermophilus suslicus. Genetic variability and differentiation for the forms with different chromosome sets were estimated. Two discriminative loci (Alb and Tf) for the studied chromosome forms were found. The UPGMA dendrogram was constructed, which summarizes genetic (allozyme) relationships found for the forms of the spotted souslik with different chromosome sets. Subdividing the species into two karyotypic forms was shown to be followed by differentiating these forms at the allozyme level.

[Variability of cytogenetic characteristics of the spotted ground squirrel Guld. 1770]. / Izmenchivost' tsitogeneticheskikh kharakteristik krapchatogo suslika Spermophilus suslicus Guld. 1770.

Females with abnormal karyotype have been found during studies of genetic variability in the Spermophilus suslicus application of the methods of differential staining has shown that the chromosome aberration are due to deletions of heterochromatin arms. Possible causes of chromosome in the S. suslicus are discussed.

Phylogenetic relationship between the Iriomote cat and the leopard cat, Felis bengalensis, based on the ribosomal DNA.

We analyzed the restriction fragment length polymorphisms in the spacer regions of ribosomal DNA (rDNA), using twelve restriction enzymes, to examine whether the Iriomote cat is related to the leopard cat (Felis bengalensis). A restriction map for each taxon was constructed and the major taxon-specific types of repeating unit (repetypes) were characterized on the basis of the arrangements of restriction sites. The Iriomote cat and the leopard cat share a common repetype but this repetype is different from that of the domestic cat (F. catus) with an estimated sequence divergence of 1.5% and from that of the ocelot (F. paradalis) with an estimated sequence divergence of 2.5%. These results indicate that, phylogenetically, the Iriomote cat is closely related to the leopard cat and that the ancestral population moved from the continent to Iriomote Island quite recently. The rDNA arrays of the leopard cat exhibit considerable intragenomic size-variation, which is thought to have emerged as a result of differences in numbers of repeated DNA segments, whereas the extent of such size-variation is much lower in the rDNA of the Iriomote cat. It appears that, even though migration of the Iriomote cat occurred relatively recently, the population has diverged to some extent from its continental counterpart, perhaps via fixation of preexistent intraspecific variations rather than by generation of new variations.

[Detection of tandem repeats in the genome of Citellus genus using restrictases]. / Obnaruzhenie tandemnykh povtorakh v genomakh suslikov roda Citellus s pomoshch'iu restriktaz.

DNA from ground squirrels of the Citellus genus (Rodentia, Sciuridae) were analysed by centrifugation in the presence of CsCl followed by digestion by restriction endonucleases. Digestion of DNA of two species C. undulatus and C. fulvus by 10 of the 16 restriction endonucleases used led to formation of electrophoretically discrete fragments that are multiple to 330 b.p. in length which points out the tandem organization of repetitive sequences similar to the satellite DNA of many mammal species. However, upon centrifugation we failed to reveal a satellite band in these species; hence the tandem repeats refer to the class of cryptic satellites in the ground squirrels and do not differ in base composition from the remaining part of DNA. The main fraction of the genome was revealed in the form of discrete fragments by cleavage with HindIII and AluI. Both of these restriction endonucleases were used for comparative analysis of DNA of 12 Citellus species. It has been shown that DNA of all species can be digested by HindIII and yields a series of fragments that are multiple to 330-30 b.p. in length and the total content of which varies from species to species within 4-22%. The fraction of the tandem repeats does not correlate with the systematic position of species nor with the amount of heterochromatin in the chromosomes. AluI cuts the DNA of 11 species yielding 110 and 220 b.p. fragments compared to only 60 and 280 b.p. in the DNA of C. dauricus. Under HindIII digestion we can also reveal the tandem repeats in marmot, which is phylogenetically close to the Citellus of the Marmota genus, but they have another periodicity--180 b.p. We propose that the age of ground squirrels repeats is 2-3 million years and they are significantly younger than the marmot repeats.