[Genetic diversity of Anser albifrons Scopoli, 1769 and Anser fabalis Latham, 1787 in the Russian Far East].

Using RAPD PCR analysis and sequencing of the 5' end segment of the mtDNA control region, the genetic diversity and differentiation of Far Eastern populations of greater white-fronted goose Anser albifrons Scopoli, 1769 and bean goose Anser fabalis Latham, 1787 were examined. Based on RAPD PCR data, the level of gene diversity (h) for A. albifrons (0.3634) and A. fabalis (0.3899) was calculated. Sequence data showed considerably higher level of inter-population diversity in A. fabalis (26.4%), compared to A. albifrons (1.88%). Similarly, the nucleotide and haplotype diversity parameters were somewhat higher in A. fabalis (0.01852 and 0.955). Phylogenetic reconstructions generated using neighbor-joining and maximum parsimony algorithms divided each of the examined species into two clusters that differ in the number of haplotypes included. These clusters can correspond to the subspecies that live in the Far East.

[Phylogeography of the mallard Anas platyrhynchos from Eurasia inferred from sequencing of the mtDNA control region].

Phylogenetic relationships, demographic history, and geographic distribution of the mtDNA haplotypes of the mallard Anas platyrhynchos were examined in three populations, Indian, Northern European, and Far Eastern. Two divergent haplotype groups, A and B, were found in the Far Eastern population, while haplotypes identified in Northern European and Far Eastern populations were exclusively of the A group. The presence of B group haplotypes in the Far Eastern population can be explained either in terms of hybridization of the mallard with spot-billed duck Anas zonorhyncha at the south of the Russian Far East, or by the mtDNA paraphyly in mallards. In general, mallards from Eurasia were characterized by low genetic population differentiation along with slightly expressed phylogeographic structure. The most differentiated was the population from India (Phi(st) = 0.076-0.077), while the difference between Northern European and Far Eastern populations was extremely low (Phi(st) = 0.0029). Differentiation of Anas platyrhynchos Indian population was determined by the fact that a part of the population, inhabiting southern and eastern coasts of the Hindustan Peninsula, was resident.

[Population genetic structure of wild-growing ginseng (Planax ginseng C. A. Meyer) assessed using AFLP markers].

Genetic variability in ten populations of wild-growing ginseng was assessed using AFLP markers with the application of fragment analysis on a genetic analyzer. The variation indices were high in the populations (P = 55.68%; H(S) = 0.1891) and for the species (P = 99.65%; H(S) = 0.2857). Considerable and statistically significant population differentiation was demonstrated (theta = 0.363; Bayesian approach, "full model"; F(ST) = 0.36, AMOVA). The results of AMOVA and Bayesian analysis indicate that 64.46% of variability is found within the populations. Mantel test showed no correlation between the genetic and geographic distances among the populations (r = -0.174; P = 0.817). Hierarchical AMOVA and analysis of genetic relationships based on Euclidean distances (NJ, PCoA, and MST) identified two divergent population groups of ginseng. Low gene flow between these groups (N(m) = 0.4) suggests their demographic independence. In accordance to the concept of evolutionary significant units (ESU), these population groups, in terms of the strategy and tactics for conservation and management of natural resources, should be treated as management units (MUs). The MS tree topology suggests recolonization of southern Sikhote-Alin by ginseng along two directions, from south and west.

[Genetic diversity of a rare species Aristolochia contorta bungae (aristolochiaceae) in Primorsky Krai].

The herbaceous vine, twisted birthwort Aristolochia contorta Bunge, is a rare species listed in the Red Data Book of Primorsky krai (2008). On the northern boundary of its range (south of the Russian Far East), the species is represented by small isolated populations confined to the river drainages. Using allozyme analysis, genetic variation of nine natural populations ofA. contorta (247 accessions), which represented the main part of the species range in Russia, was examined. The values of genetic variation indices (P = 22.7%; A = 1.28; Ho = 0.129; He = 0.101) were low and comparable with the data obtained for other rare plants. The proportion of unique genotypes (G/N) and Simpson's genotypic diversity index (D) ranged from 0.32 to 0.64 and from 0.60 to 0.98, respectively. This means that A. contorta is characterized by sexual and asexual reproduction. Moreover, the ratio between these types of reproduction varied among the populations. Complete absence of inbreeding and excess of heterozygotes (F(IS) = -0.282), which was low probable in case of free mating, was observed. Evidently, clonal growth and (or) apomixis enables the species to maintain certain level of heterozygosity despite of small population sizes and non-regular gene exchange.

[Heterogeneity of the spatial distribution of the primordial organic substance as an initial stage of biological evolution].

An approach to describe the emergence of the primordial evolving system has been developed. The dynamics of polymerization/depolymerization of some spatially distributed prebiological structures has been analyzed, and two phases of the development of the system have been identified. In the first phase, the polymerization of organic monomers occurs by the influence of external factors, and in the second one depolymerization takes place. Both processes are accompanied by "diffuse mixing" of reaction products. The dynamic equations of the system are presented. The numerical examination of the space nonuniform solution of model equations has shown that, in conditions of low stability of uniform space distribution, these solutions resolve into a number of discrete peaks of non-zero density, which are isolated from each other by free space. Such nonuniform distributions are stable when being close to the bifurcation point; yet in other conditions, they can lose their stability, which entails a more pronounced nonuniformity of space dynamics. Thus, interaction of polymerization/depolymerization processes results in the chaotic self-organization and leads to the origination of complex and nomhomogeneous (putchy) spatial structures. These structures can reflect the emergency of the spatial nonuniformity in primordial associations, in physical space, in the distributive space of characters can correspond to the initial steps of individualization of the first discrete domains fixed in the biological evolution.

[Polymorphism of RAPD, ISSR and AFLP markers of the Panax ginseng C. A. Meyer (Araliaceae) genome].

The genus Panax (Araliaceae) is world-famous because many its members have important medicinal properties. Panax ginseng C. A. Meyer is more popular than other species of the genus because remedies prepared from this plant stimulate immunity, help to prevent diseases, and have antistress effects. In addition, the ginseng root extract is traditionally used as a means against aging. At present, this species is found in the wild only in Primorsky krai, Russia, but its populations are extremely exhausted and need to be restored. In this study, effectiveness of molecular DNA markers in detecting genetic variation and differentiation of the ginseng populations was tested. Genetic variation of ginseng, identified using RAPD (P = 4%; H(pop) = 0.0130) and ISSR (P = 9.3%; H(pop) = 0.0139) markers was low. The AFLP* approach, according to which amplicons are separated in polyacrylamide gel and visualized by means of silver staining, showed somewhat higher variability (P = 21.8%; H(pop) = 0.0509), while its effectiveness in population differentiation was as low as that of RAPD and ISSR. The AFLP** technique, which included analysis of the fragments using genetic analyzer, revealed high genetic differentiation of ginseng (P = 94.4%; H(pop) = 0.3246). All populations examined using the AFLP** markers were statistically significantly differentiated based on the AMOVA results. Our result suggest effectiveness of AFLP** markers for characterization of the genetic structure and genetic relationships of the ginseng populations. These markers are recommended for use in large-scale population genetic studies of this species to develop measures of its conservation.

[Genetic structure of the Far Eastern population of Eurasian wigeon Anas penelope inferred from sequencing of the mitochondrial DNA control region].

Sequence variation of the 5' end of the mitochondrial DNA control region (600 bp) was examined in the population samples of Eurasian wigeon Anas penelope from Anadyr' and Primorye. A total of 11 different mtDNA haplotypes were identified, with one of these belonging to American wigeon Anas americana. The presence of the mtDNA haplotype from the species closely relative to A. penelope in the Anadyr' sample can be considered as the genetic evidence in favor of interspecific hybridization. This suggestion is in the good agreement with ornithological data. Genetic differentiation of the Primorye and Anadyr' populations was low (phi(ST) = 0.096). The phylogeographic structure was not pronounced.

[Genetic variation of Oplopanax elatus (Nakai) Nakai (Araliaceae)].

Using allozyme analysis, genetic and genotypic variation of rare relict species, a member of one of ancient angiosperm families, Araliaceae, the clone-forming plant Oplopanax elatus (Nakai) Nakai, was evaluated. Electrophoretic separation of the enzymes is described, and genetic interpretation of the enzymes variation patterns is presented. The values of genetic variation indices obtained were low (P = 25%; A = 1.45: Ho = 0.131; He = 0.113) and comparable with the data reported for the rare plant species and the representatives of the family Araliaceae. The main factors responsible for the polymorphism level observed might be the evolutionary history of the species and gene drift. The level of genotypic diversity (G/N = 0.76; D = 0.97) was substantially higher, compared to the values reported for species with vegetative reproduction (D = 0.62). These results suggest certain contribution of the propagation by seeds in the formation of the species of interest.

[Polymorphism of a short fragment of the mitochondrial genome control region (D-loop) in the Siberian roe deer Capreolus pygargus Pallas, 1771 (Artiodactyla, Cervidae) from the Russian Far East].

Genetic diversity of Siberian roe deer Capreolus pygargus, 1771 from the Russian Far East was studied based on polymorphism analysis of a mtDNA control region fragment (390 bp). Three phylogenetic lines were found in the animals examined. The trend for change of haplotypes of different phylogroups was shown to occur from north to south in Sikhote-Alin. The haplotype distribution of Siberian roe deer in the Russian Far East correlated with data on morphologiocal variability.

[Larch chromosomal mosaicism].

Karyological study of larch population members from different parts of the Larix olgensis L. Henry areal in the Primorskii krai has been carried out. The main amount of chromosomes for larch as n = 12 (2n = 24) has been confirmed. Mixoploidy was observed in all studied populations. The difference of individuals from the larch areal based on the cell amount with a different level of ploidity has been found in the population of L. olgensis locus classicus and both L. sibirica and L. gmelinii, which is probably a consequence of their hybrid nature.

[Characteristics of gene pools of three Pinus pumila (Pall.) Regel populations at the range margins].

Genetic variation of Siberian dwarf pine Pinus pumila (Pall.) Regel was characterized in three marginal populations in southwestern, southern and eastern parts of the natural species range (Transbaikalia, Primorye, Kamchatka) using isozyme analysis. Analysis involving 16 isozyme loci encoding ten enzyme systems was conducted. Our results confirm that P. pumila is among the most polymorphic species in the family Pinus. Three marginal populations exhibited high genetic variation (P95 = 68.8%, Ho = 0.247, He = 0.291). Populations heterogeneity and significantly high level of divergence in coniferous (F(ST) = 0.050, D(N) = 0.044) reflect their genetic originality. In summary, it was shown that the level of genetic variation characteristic for P. pumila in other parts of the not only is reproduced in the populations examined but even is close to maximum there.

[Genetic structure of natural populations of the relict species Aristolochia manshuriensis (Aristolochiaceae) in disturbed and intact habitats].

Genetic structure of natural populations of the rare relict plant Manchurian birthwort (Aristolochia manshuriensis Kom.) in the Russian part of its area was analyzed using allozyme markers. The studied A. manshuriensis populations differed in the degree of their intrapopulation differentiation. The populations Nezhinka and Anan'evka were more differentiated (F(ST) = 0.1209 and 0.0576, respectively); these populations are located close in the regions of intense economic activity and are exposed to the strongest anthropogenic impact. A low degree of differentiation was detected in the population Malaya Borisovka (F(ST) = 0.0393), localized to intact habitats. The overall heterogeneity test has demonstrated that the population Malaya Anan'evka, exposed at present to small anthropogenic stress yet growing in disturbed habitats, displays no differentiation. These results suggest that at least three populations are influenced by genetic drift connected with a decrease in the reproductive and effective population sizes, which is caused, in particular, by anthropogenic impact. A high level of genetic similarity between the A. manshuriensis populations in Primor'e is discussed in connection with the evolutionary history of this species.

[Genetic variation of Manchurian pheasant (Phasianus colchicus pallasi Rotshild, 1903) inferred from mitochondrial DNA control region sequences].

Sequence variation of the mitochondrial DNA control region was studied in Manchurian pheasants (Phasianus colchicus pallasi Rotshild, 1903) representing three geographic populations from the southern part of the Russian Far East. Extremely low population genetic differentiation (F(ST) = 0.0003) pointed to a very high gene exchange between the populations. Combination of such characters as high haplotype diversity (0.884 to 0.913), low nucleotide diversity (0.0016 to 0.0022), low R2 values (0.1235 to 0.1337), certain patterns of pairwise-difference distributions, and the absence of phylogenetic structure suggested that the phylogenetic history of Ph. C. pallasi included passing through a bottleneck with further expansion in the postglacial period. According to the data obtained, it was suggested that differentiation between the mitochondrial lineages started approximately 100 000 years ago.

[Genetic structure and differentiation of populations of the tetraploid species Oxytropis chankaensis (Fabaceae)].

The population genetic variation of the tetraploid species Oxytropis chankaensis Jurtz. (Fabaceae), a local endemic of the western coast of Khanka Lake (Primorye), was examined. Five populations were analyzed using 28 isozyme loci encoding 16 enzyme systems. Significant allelic heterogeneity among the populations was found for six out of twelve polymorphic loci. The heterozygosity of the samples (total sample size 294 plants) He = 0.301 was considerable higher than the mean values in populations of endemic species (0.076). Based on the results of this study, we identified two groups of O. chankaensis populations (southern and northern), in spite of the absence of marked hiatus between them. Of special interest is the population from Przhewalski Spit, which is a natural reserve of genetic diversity of the species and the putative center of formation of the autotetraploid O. chankaensis.

[Independent species status of Iris vorobievii N.S. Pavlova, Iris mandshurica Maxim., and Iris humilis Georgi (Iridaceae): evidence from the nuclear and chloroplast genomes].

Nuclear (RAPD) and chloroplast (trnH-psbA, atpB-rbcL, rps4, trnL-trnF, and trnS-trnG) molecular DNA markers were used to study the populations of closely related species of the genus Iris L.: Iris vorobievii N.S. Pavlova, I. mandshurica Maxim., and I. humilis Georgi. An analysis of 243 RAPD fragments made it possible to calculate the main population parameters and to identify species-specific RAPD markers. Differences that discriminated the species were found in all but one (trnL-trnF) cpDNA region. The total length of the four variable cpDNA regions was 3640 bp, indels included. Their variation was low. A total of 22 variable sites (0.63%) were found; of these, 13 were parsimony informative and 9 were noninformative. Analysis of molecular variance (AMOVA) revealed significantly (P < 0.0001) high differentiation of both nuclear (F(ST) = 0.681) and chloroplast (F(ST) = 0.854) genomes for the species examined. Thus, a multigene approach confirmed the independent species status of I. vorobievii, I. mandshurica, and I. humilis and showed that I. vorobievii and I. mandshurica, but not I. humilis, are found in Primorye.

[Specific features of pollination in the Manchurian birthwort Aristolochia manshuriensis].

An analysis of pollination system in Aristolochia manshuriensis has shown that flower structure in this species is strictly adapted to cross-pollination, but the possibility of an autogamous or geitonogamous type of self-pollination with the involvement of insects is not excluded. The flowers of A. manshuriensis are most frequently visited by flies of the family Anthomyiidae, which markedly contribute to their pollination. Males account for 65% of pollinator insects collected from the flowers.

[Calcium-dependent mechanism of somatic embryogenesis in oncogene rolC expressing cell cultures of Panax ginseng].

It was shown earlier, that ginseng embryogenic cell culture 2c3 was obtained as a result of callus cells transformation with the Agrobacterium rhizogenes rolC oncogene. In the present report we determine that inhibitors of Ca2+-channels (LaCl3, verapamil, niflumic acid) certainly lowered the quantity of somatic embryos in the 2c3 cell culture. This is the evidence of the influence of calcium-dependent signal system on plant embryogenesis. Protein kinases inhibitors W7 and H7 also caused the lowering of somatic embryos quantity in the 2c3 cell culture. We analysed changes of CDPK genes expression in embryogenic 2c3 cell culture. Total expression decreased 1.2-1.5 times comparing with the control callus culture. CDPK expression in the 2c3 embryogenic culture lowered by the inhibition of expression of the gene subfamilies PgCDPK1 (PgCDPK1a and PgCDPK1b) and PgCDPK3 (PgCDPK3a). At the same time, expression of PgCDPK2 gene subfamily (PgCDPK2b and PgCDPK2d) was increased. We suppose that genes of PgCDPK2 subfamily might be responsible for the embryogenesis initiation in the 2c3 ginseng cell culture. It was shown for the first time that the rolC gene and the process of embryogenesis could change expression of particular forms of CDPK genes.

[Genetic variability and differentiation in the larch populations within the range of Larix olgensis A. Henry in Primor'e region].

The level of within- and among-population variation of larch inhabiting the range of Larix olgensis A. Henry in Primorye was estimated on the basis of 440 RAPD loci identified by means of 12 random primers. In ten population examined, the proportion of polymorphic loci was 35-60%, the average expected heterozygosity varied from 0.1340 to 2169, and the average gene flow estimate was 1.38. According to Fisher's test for heterogeneity, the pairwise differences of the fragment frequencies between the populations were statistically significant. The subdivision index G(ST) = 0.2663 indicated that the interpopulation variation component accounted for approximately 27% of the total variation. Coefficients of Nei's genetic distance between the populations varied from 0.0137 to 0.0934. Most of the samples with high support clustered according to the geographic positions relative to one another within the range. These results suggest that the populations examined are characterized by high genetic variation, like the larch populations of Siberia and the Russian Far East studied earlier, but, in contrast to the latter, exhibit higher among-population variability.

[Allozyme variation of the relict plant Aristolochia manshuriensis Kom. (Aristolochiaceae)].

Allozyme variation of a rare relict plant, birthwort Aristolochia manshuriensis Kom., was examined. The main parameters of genetic variation in natural populations of A. manshuriensis from the Anan'evka, Nezhinka, and Malaya Borisovka river basins (Primorskii Krai) were inferred from analysis of nine enzyme systems, presumably encoded by 18 loci. At 99% polymorphism criterion, 24.4% of A. manshuriensis loci were shown to be polymorphic. The mean number of alleles per locus was 1.24; the mean observed and expected heterozygosities, 0.12 and 0.10, respectively.

[Phylogenetic relationships of the Far Eastern Araliaceae inferred from ITS sequences of nuclear rDNA].

In eight species of the family Araliaceae, inhabiting the territory of the Russian Far East, the sequences of ITS regions of nuclear rDNA were determined. A comparison of these sequences enabled establishment of phylogenetic relationships between the Far Eastern and other members of the family. It was demonstrated that Aralia sensu populations from Primorye and Sakhalin were genetically different and, hereby, could be classified as interspecific taxa. Aralia continentalis along with A. cordata were attributed to the section Aralia sensu. Oplopanax elaus and O. horridus were found to be very close to each other, possibly being the subspecies of one species or relatively young species. Legitimacy of the isolation of two sections within the genus Eleutherococcus was confirmed.