[Geographical structure of the sable (Martes zibellina L.) gene pool on the basis of microsatellite loci analysis].

The genetic structure of seven natural sable populations was investigated with the use of the original panel of 10 microsatellite loci. The populations were selected on the basis of the historical data on sable numbers fluctuations for the last 300 years, as well as on data on natural and artificial migrations affecting neighboring populations. We have demonstrated that the populations are in a state of genetic equilibrium for the majority of the loci. The genetic differences between three samplings from Central Siberia populations were insignificant, and the fixation index values were relatively low. At the same time, populations from the margins of the species habitation areal were characterized by the highest fixation index values. We have shown for the first time that populations from different regions of the sable habitation areal maintain their specific features despite the influence of natural and artificial migrations. The current study, performed with nuclear genetic markers, made it possible to get insight into the genetic structure of the analyzed species as a whole.

[The 3'-UTR polymorphism of dopamine transporter gene in hadza and datoga males].

The study of VNTR-polymorphism and the molecular struc ture of 3'-UTR of the dopamine transporter gene (DAT1/SLC6A3) was performed in hadza and datoga males. It was shown that hadza and datoga differed in allele and genotype frequencies. Allele with 9 repeats in 3'-UTR is more common in hadza as well as homozygous genotype DAT19/9. Allele with 10 repeats is more common in datoga as well as homozygous genotype DAT1 10/10. The molecular structure of the DAT alleles with 3, 8 and 12 repeats was determined for the first time. In addition it was found that DAT1 allele with 11 repeats in datoga significantly differed from previously described ones in other populations in repeats type and arrangement. We suggest that variations of the repeats num ber and type in the 3'-UTR of allelic variants may affect the dopamine transporter gene function.

[Polymorphism of 5-HTTLPR and Stin2 loci of the serotonin transporter gene in males of African ethnic populations Hadza and Datoga].

A molecular-genetic study of 5-HTTLPR and the Stin2 loci of the serotonin transporter gene (5-HTTL) in males of the African ethnic populations Hadza and Datoga, which differ in the level of culturally acceptable aggression, was carried out. The distribution of allele and genotype frequencies of these two loci was established. It was shown that the frequency distribution of genotypes and alleles among Hadza and Datoga in the examined samples is practically identical by the VNTR-polymorphism of both loci. However, the Hadza populations, as compared to Datoga, showed a significant (p = 0.006) increase in the frequency of the transcriptionally less active allele L(G) of the 5-HTTLPR locus. For the first time, the structure of the allelic variant of locus Stin2 with eight repetitions (Stin2.8) is described and established for African populations. The test for independence of the frequency distribution of the alleles of the studied loci showed highly significant linkage disequilibrium among Hadza (p << 0.001) and Datoga (p = 0.021). In analysis of the genotype combinations of two loci, it was revealed that thestudied populations differed significantly by the L(A)L(G) 10/12 genotype (p << 0.001). When combining the genotypes, no significant differences between the populations based on their expression activity were identified. We assume that the identified combined genotypes reflect the effects of similar behavioral traits for both populations.

[Interspecies variability of number of bristles on dorsal surface of aedeagus in D. virilis species group and its genetic mapping with interspecies hybrids of D. virilis and D. lummei].

The results of morphologic and hybrid analyses of the feature of the male reproductive system of sibling species in the virilis group were presented. Bristles appeared on the surfaces of male genitals (aedeagus). The occurrence of a specific expression of the examined feature in the phyllades of D. virilis group, the correspondence of both the number and distribution pattern of the bristles on surfaces ofthe aedeagus and developmental temperature in D. virilis and D. lummei, as well as the link between feature and sexual behavior, have been shown. Dominance of D. lummei phenotype in the interspecies D. virilis x D. lummei was found. The interspecies hybrids D. virilis and D. lummei were used for a genetic analysis of the variability of the examined feature. The significant influence of chromosomes 2 and 6 on the number of bristles on the aedeagus in hybrid males was shown. Furthermore, the correspondence between the effects of the autosomes 2 and 6 on the variability of the examined feature and the genetic status of the other chromosomes (the effect of interaction between genetic factors, chromosomes here) was revealed. The adaptive value of the examined feature related to the involvement in the formation of isolating barriers at the copulation stage is under discussion.

[Genetic structure of sable (Martes zibellina L.) from Eurasia based on distribution of mitochondrial lineages].

The phylogeography of the sable, which is a commercially valuable species, is extremely complicated and poorly investigated. Specifically, the effects of factors such as the range dynamics of the sable during the Pleistocene Epoch, the localization of glacial refugia, species distribution pattern in Holocene, and recent dramatic population decline, along with massive reacclimatization measures, on the species phylogeography remain unclear. Based on the sequence analysis of the control region of mitochondrial DNA from sables that inhabit different parts of the species range, a suggestion was made of the considerably high Pleistocene genetic diversity in sable, which was subsequently lost. The initial diversity of mitochondrial lineages is mostly preserved in the Urals, while in the eastern part of the range, it seems to have been depleted as early as before the last glacial maximum. On the other hand, the even greater depletion of the mitochondrial lineages observed in some populations of central Siberia can be associated with the dramatic population decline at the turn of the 20th century.

[Molecular genetic polymorphism of androgen receptor gene (AR) in African populations of Hadza and Datoga].

The molecular genetic analysis of the polymorphic variants of the CAG repeat-containing locus of the andro- gen receptor (AR) gene was performed in the populations of Hadza and Datoga. Allele frequency distribution patterns were established. Alleles containing 20-25 repeats were the most abundant in both populations were the. The populations studied were compared with Asians (Han), white Americans, and Africans (Ariaal). Sta- tistically significant difference between populations of Hadza and Datoga in the distribution of the AR allelic variants was demonstrated.

[Polymorphism of the dopamine D4 receptor (DRD4) and serotonin transporter (5-HTTL) gene promoter regions in African tribes of Hadza and Datoga].

Molecular genetic analysis of the allelic variants of the DRD4 and 5-HTTL gene promoter regions was performed in African tribes of Hadza and Datoga, characterized by different levels of socially acceptable aggression. It was demonstrated that Hadza and Datoga people differed in the structural organization of one of the 5-HTTL alleles (extra long allele xL). Analysis of the allele length polymorphism of both genes showed that in the Hadza and Datoga samples examined, variation parameters, as well as the genotype and allele frequency distribution pattern were almost the same. At the same time, analysis of the SNP polymorphism at the A/G substitutions of the 5-HTTL locus revealed a substantial decrease of the active allele L(A) frequency in the population of Hadza compared to the population of Datoga (chi2 = 3.77; d.f = 1; P = 0.052).

[Microsatellite analysis of two captive populations of sable (Martes zibellina L.)].

The high value of sable (Martes zibellina L.) fur and stable demand for it over the centuries have led to suboptimal hunting patterns and, as a result, considerable fluctuations in the sizes of natural populations of this species. To maintain the traditional export of sable fur, efforts towards commercial domestication of sable have been made in Russia. The first farm population of sable consisted of animal from eight natural populations in 1929. After the problems related to breeding in captivity were solved, directional selection began. Eighty years of breeding have resulted in sable herds with homogeneous quantitative characters. Prospects for further breeding depend on the current level of genetic diversity in the captive populations of sables formed during the first stages of domestication. The sable populations of the Pushkinsky and Saltykovsky fur farms located in Moscow oblast, which were the objects of this study, are the progenitors of the existing captive populations. The first estimation of genetic variation of this species by means of a panel of microsatellite markers was developed for this study. Two captive sable populations were analyzed using ten microsatellite loci; a total of 75 alleles were found in both populations. Population-specific alleles were identified (6 and 13 in the Pushkinsky and Saltykovsky populations, respectively). The populations studied were found to be differentiated with respect to four microsatellite loci.

[The use of the ISSR-PCR method for identifying domesticated animal breeds and species, inferring their population structures, and assessing gene pool similarity].

The use of multilocus intermicrosatellite assay (ISSR-PCR) in combination with the method of k-clustering of population structures (Structure v2.2. program) has become a basis for a battery of tests of molecular genetic examination and for studying the gene pool of domesticated animal species. Possibilities are reviewed for analysis of the population structures of domesticated species at different levels (interspecific, interbreed, and intrabreed levels), for assessment of similarity of gene pools of breeds (intrabreed groups), as well as for identification of breeds and estimation of their consolidation, purity, and genealogical relationships.

[Study of genetic variation in Yakutian cattle (Bos taurus L.) using the prolactin bPRL, growth hormone bGH, and transcription factor bPit-1 genes].

The genetic structure of the Yakutian cattle breed was studied using the following genes: bPRL (RsaI site in exon 3), bGH (AluI site in exon 5), and bPit-1 (HinfI site in exon 6). The values of observed heterozygosity were 0.36 for bPRL, 0.29 for bGH, and 0.16 for bPit-1. These values are within the range of values for this parameter established for a number of Bos taurus breeds. The results obtained show that genetic variation is preserved in this aboriginal Russian breed, despite a catastrophic reduction of the number of animals.

[Nonuniformity in the evolutionary rate in the virilis: II. group of Drosophilas: application of the method of Tajima's test].

The steadiness of the molecular clock was estimated in 11 Drosophila species of the virilis group by sequences of five genes by applying Tajima's Simple Method. The main characteristic of this method is the independence of its phylogenetic constructions. The obtained results have completely confirmed the conclusions drawn relying on the application of the two-cluster test and the Takezaki branch-length test. In addition, the deviation of the molecular clock has found confirmation in D. virilis evolutionary lineages.

[Irregular evolutionary rate of the fruit fly from the Virilis group: I. Use of phylogeny-mediated Takezaki tests].

The aim of this study is to test the hypothesis about an inconsistency in time estimation of the divergence and topology of species Drosophila kanekoi, D. ezoana, and D. littoralis which is caused by the irregular accumulation rate of replacements in different DNA sequences used for analysis and in evolutionary lineages. The phylogenetic relationships based on sequences of five genes among 11 fruit fly species of virilis group are revised, and estimation of molecular clock regularity in several phylogenetic lineages of this group is given. It was shown that Drosophila kanekoi, D. ezoana, and D. littoralis assembled in a single cluster, which was most related to the subfilade montana. The irregularity of molecular clocks was shown for the high conservative sequences of mt 12S-16S.

[The effect of artificial selection for coat color on fitness in a farm population of the sable (Martes zibellina)].

The relationship between the response to artificial selection for darker coat color and fitness in a farm population of the sable (Martes zibellina L.) from the Pushkinskoe Fur Farm (Moscow oblast) was studied. The selection was performed during 41 years. By the moment of the study, a response to the selection for this character had been obtained: the coat color in the selected population had become darker, and the proportion of black animals in it increased. In addition, sables with black heads, which were absent in the original population, had appeared. Artificial selection was accompanied by a decrease in the fitness of the selected population, which was expressed in decreased female reproductive capacity parameters (the fertility, maturation rate, and duration of the reproductive period). A selection technique consisting in the use of only highly fertile animals in the selection originally made it possible to restore the fitness parameters to the initial level almost without a decrease in the dark shade of the fur. However, further selection led to a drastic decrease in fitness that could not be precluded by any selection method used. The possible ways to overcome this unfavorable effect of artificial selection are discussed.

[The relationships among the species of the Drosophila virilis group inferred from the gene Ras1 sequences].

Comparative analysis of a group of closely related Drosophila species (D. virilis, D. lummei, D. novamexicana, D. americana texana, D. flavomontana, D. montana, D. borealis, D. lacicola, D. littoralis, D. kanekoi, and D. ezoana) was conducted based on an incomplete sequence of gene Ras1. The pattern of the relationships among the species corresponded to that expected from analysis of morphological and cytogenetic characters. Statistical data favoring neutrality of the substitutions examined in the Ras1 gene are presented. This character of the gene Ras1 evolution confers more reliability to reconstruction of phylogenetic relationships among closely related species. The resultant tree for main phylads of the group is as follows: (D. virilis, D. lummei, D. montana, D. ezoana).

[The endosymbiotic bacterium Wolbachia enhances the nonspecific resistance to insect pathogens and alters behavior of Drosophila melanogaster].

To determine biologically important effects of the cytoplasmic endosymbiont Wolbachia, two substrains of the same Drosophila melanogaster strain have been studied, one of them infected with Wolbachia and the other treated with tetracycline to eliminate the bacterium. Female D. melanogaster infected with Wolbachia are more resistant to the fungus Bauveria bassiana (an insect pathogen) than uninfected females; infected females also exhibited changes in oviposition substrate preference. Males infected with the bacterium are more competitive than uninfected males. The possible role of Wolbachia in the formation of alternative ecological strategies of D. melanogaster is discussed.

Intraspecific variation and population structure of the German cockroach, Blattella germanica, revealed with RFLP analysis of the non-transcribed spacer region of ribosomal DNA.

Little information is available on genetic variation within and between populations of pest cockroaches. In this study, intraspecific HindIII polymorphism was investigated in the German cockroach, Blattella germanica (Linnaeus) (Dictyoptera, Blattaria: Blattellidae), using restriction fragment length polymorphisms (RFLP) of the non-transcribed spacer (NTS) region of ribosomal DNA (rDNA). Individual male insects were collected from infestations at three different pig farms. Each population was characterized by HindIII restriction fragment frequencies and haplotype (a particular X-chromosome pattern) frequencies. The inheritance of the X-chromosome HindIII rDNA patterns over 12 generations (3 years) follows Mendelian patterns, and the stability of this polymorphic marker indicates infrequent genetic recombination of variable sites. Although pairwise genetic distance measures were uncorrelated with geographical distance, the pattern of genetic differentiation of the three cockroach populations suggests that human-mediated transport of cockroaches is an important force in shaping the population genetic structure of cockroach infestations, at least at the regional scale of 10-100 km. Sequence variation in the ribosomal NTS is a useful marker, and RFLP of rDNA is a simple, robust and reproducible technique for differentiating recently diverged cockroach populations.

[Allelic polymorphism of kappa-casein gene (CSN3) in Russian cattle breeds and its informative value as a genetic marker].

The frequencies of the kappa-casein gene (CSN3) alleles and genotypes have been determined in five Russian cattle breeds (Bestuzhev, Kalmyk, Russian Black Pied, Yaroslavl, and Yakut breeds) by means of PCR-RFLP analysis using two independent restriction nucleases (HinfI and TaqI) and by allele-specific PCR. Typing alleles A and B of CSN3 is of practical importance, because allele B is correlated with commercially valuable parameters of milk productivity (protein content and milk yield) and improves the cheese yielding capacity. The frequencies of the B allele of CSN3 in the breeds studied vary from 0.16 to 0.50; and those of the AB and BB genotypes, from 0.27 to 0.60 and from 0.02 to 0.23, respectively. The Yaroslavl breed had the highest frequencies of CSN3 allele B and genotype BB (0.50 and 0.23, respectively). The frequencies of the B allele and BB genotype in other breeds studied varied from 0.25 to 0.32 and from 0.03 to 0.09, respectively. In none of the breeds studied have the observed and expected heterozygosities been found to differ from each other significantly. However, the observed genotype distributions significantly differ from the expected one in some herds (in most such cases, an excess of heterozygotes is observed). Two herds of the Yaroslavl breed dramatically differ from each other in the heterozygosity level: a deficit (D = -0.14) and an excess (D = 0.20) of heterozygotes have been observed at the Mikhailovskoe and Gorshikha farms, respectively. In general, however, the heterozygosity of the Yaroslavl breed corresponds to the expected level (D = 0.04). Analysis of breeds for homogeneity with the use of Kulback's test has shown that all cattle breeds studied are heterogeneous, the CSN3 diversity within breeds being higher than that among different breeds, which is confirmed by low Fst values (0.0025-0.0431). Thus, a DNA marker based on CSN3 gene polymorphism is extremely important for breeding practice as a marker of milk quality; however, it is inapplicable to marking differences between breeds or phylogenetic relationships between cattle breeds because of the high diversity with respect to this locus within breeds.

[A RAPD fingerprinting of sibling species of the Drosophila virilis group].

A comparative analysis of the sibling species of Drosophila virilis was performed by RAPD-PCR technique using a set of random primers. The degree of relatedness was studied by cluster analysis (UPGMA) and multi-dimensional scaling. The resulting pattern of species relationships contradicts the classical taxonomy. The main result of the cluster analysis is that D. virilis does not cluster with the remaining three species of its phylad, while according to multidimensional scaling, D. virilis is equidistant from all the species of its group, from both the species of its phylad and the species of the montana phylad. The montana phylad is extremely heterogeneous; moreover, the species D. littoralis, D. ezoana, and D. kanekoi appear to be closer to the virilis phylad than to the other species of the montana phylad, wherein these species are traditionally included. The phylogenetic relationships between the studied species discovered using RAPD fingerprinting comply with the results obtained using protein markers and quantitative traits.

[Analysis of secondary sex ratio in the fox (Vulpes vulpes L.)].

Secondary sex ratio and its variability in relation to some paratypic and genetic factors were studied in the silver fox by analysis of data obtained at the Pushkinskii fur farm (Moscow oblast) in 1980-1989. A total of 17285 whelps were examined. The mean proportion of males over the ten years of observation was 0.536+/-0.004. No effect of parent age or litter size on this proportion was found. Individual analysis of the progeny of a single parent revealed 44 males and 49 females showing significant deviations from the expected sex ratio (l male : 1 female). These results can be used for reconstruction of pedigrees whose progeny yields regular deviations from the expected sex ratio.

[Identification of individual genotype parameters, informative for medical care in piloted space missions: modern and future capabilities].

The factors of space flight provoke some negative shifts in crew health. Improvement of the methods of diagnostics, evaluation of predisposition to diseases and correction of functional deviations is within the scope of the crew medical operations personnel. It is also a new focus of biomedical research referred to as "personalized medicine" the corner stone of which is genetic analysis. Investigations in this area are prediscovery by character; population genetics seems to be the most adequate approach. The authors give examples of candidate genes the genotypes of which could be of significance to medical operations and discuss the genetic safety of space flight. Aside from the applied aspects, the genetic investigations of space crews are of their own concernment in understanding the genetic basis of differences in biological resistance to unhealthy environment.