D. V. Politov

The extent and meaning of hybridization and introgression between Siberian spruce (Picea obovata) and Norway spruce (Picea abies): cryptic refugia as stepping stones to the west?

Boreal species were repeatedly exposed to ice ages and went through cycles of contraction and expansion while sister species alternated periods of contact and isolation. The resulting genetic structure is consequently complex, and demographic inferences are intrinsically challenging. The range of Norway spruce (Picea abies) and Siberian spruce (Picea obovata) covers most of northern Eurasia; yet their geographical limits and histories remain poorly understood. To delineate the hybrid zone between the two species and reconstruct their joint demographic history, we analysed variation at nuclear SSR and mitochondrial DNA in 102 and 88 populations, respectively. The dynamics of the hybrid zone was analysed with approximate Bayesian computation (ABC) followed by posterior predictive structure plot reconstruction and the presence of barriers across the range tested with estimated effective migration surfaces. To estimate the divergence time between the two species, nuclear sequences from two well‐separated populations of each species were analysed with ABC. Two main barriers divide the range of the two species: one corresponds to the hybrid zone between them, and the other separates the southern and northern domains of Norway spruce. The hybrid zone is centred on the Urals, but the genetic impact of Siberian spruce extends further west. The joint distribution of mitochondrial and nuclear variation indicates an introgression of mitochondrial DNA from Norway spruce into Siberian spruce. Overall, our data reveal a demographic history where the two species interacted frequently and where migrants originating from the Urals and the West Siberian Plain recolonized northern Russia and Scandinavia using scattered refugial populations of Norway spruce as stepping stones towards the west.

Application of Microsatellite Loci for Molecular Identification of Elite Genotypes, Analysis of Clonality, and Genetic Diversity in Aspen Populus tremula L. (Salicaceae)

Testing systems for molecular identification of micropropagated elite aspen (Populus tremula L.) genotypes were developed on the base on microsatellite (SSR) loci. Out of 33 tested microsatellite loci, 14 were selected due to sustainable PCR amplification and substantial variability in elite clones of aspen aimed for establishment of fast-rotated forest plantations. All eight tested clones had different multilocus genotypes. Among 114 trees from three reference native stands located near the established plantations, 80 haplotypes were identified while some repeated genotypes were attributed to natural clones which appeared as a result of sprouting. The selected set of SSR markers showed reliable individual identification with low probability of appearance of identical aspen genotypes (a minimum of 4.8 · 10−10 and 1 × 10−4 for unrelated and related individuals, resp.). Case studies demonstrating practical applications of the test system are described including analysis of clonal structure and levels of genetic diversity in three natural aspen stands growing in the regions where plantations made of elite clones were established.

[Distribution of the genetic diversity of the Siberian stone pine, Pinus sibirica Du Tour, along the latitudinal and longitudinal profiles].

The Siberian stone pine (Pinus sibirica Du Tour) is one of the main forest-forming coniferous species of the boreal ecosystems of Western Siberia. We used the isozyme method to analyze 11 ecotypes representing the latitudinal and longitudinal profiles within the species range, including samples from the geographic boundaries of the distribution. The genetic structure of the ecotypes is described on the basis of the variability for 26 isozyme loci encoding for 16 enzyme systems. The greatest genetic diversity was observed in the taiga ecotypes in the central part of the studied area, while the ecotypes along the species range boundaries were shown to be genetically depauperized. Approximately 8.1 % of the observed genetic diversity is attributed to differences between the studied ecotypes. We detected high levels of genetic diversity for the Fest-2, Pgm-1, Sod-4, and a few other loci, as well as a correlation between allele frequencies and geographical locations of the populations. The results of multivariate analysis of allelic frequencies as well as cluster analysis allowed us to discriminate three major groups of ecotypes: north-eastern, central and south-western. In view of our results, we compare two hypotheses: one which attributes the spatial distribution of genetic variations to the selectivity for some of the polymorphic allozyme loci, and the other based in the history of the formation of the range of the Siberian stone pine.

Genetic Diversity and Relatedness in Different Generations of the Siberian Crane (Grus leucogeranus Pallas) Captive Population

Eight variable microsatellite loci were analyzed in terms of studying the genetic structure of different generations of a captive population of a rare endemic Russian species, the Siberian crane (Grus leucogeranus Pallas). It was shown that the founding population of natural origin (15 birds) is characterized by high genetic diversity (NA = 6.625, HO = 0.767, HE = 0.731) and a lack of relatedness (R = −0.079). In the total sample of descendents of the founders (122 individuals from generations F1, F1/F2, F1/F3, F2, F2/F3), this characteristic level of genetic variation is being maintained; however, we observed a decrease in allelic richness in some generations (F1/F2, F1/F3, F2). We found a low level of relatedness in the sample of descendents of the founders (F1, F1/F2, F1/F3), while the relatedness was maximal (R = 0.302) in the descendents of the breeders of the first generation. A small sample of breeders related to each other of generations F1 and F2 (eight birds) does not represent the entire gene pool of the founders of the Siberian Crane captive population. In view of this, we discuss the need to form a new genetically heterogeneous generation of breeders that would also include Siberian Cranes from the almost extinct Western Siberian population.

Gene pool state and degree of infestation by bark beetle ( Ips tipographus L.) of Norway spruce ( Picea abies L. Karst.) natural populations and planted stands in Moscow region

A comparative analysis of the gene pool state in natural populations and planted stands of Norway spruce and the degree of their infestation by the bark beetle in the Moscow region was conducted taking into account the dynamic state of communities (4 populations, 148 samples, 24 isoenzyme loci). The degree of infestation by the bark beetle of conditionally native communities is 0%; for planted stands, it is 90–100%; and for a short-term community, it is 15–20%. The comparison of “healthy” populations and those infested with bark beetle by average values of observed heterozygosity (HO) detected no significant differences. However, the test on allelic frequency heterogeneity demonstrated the difference of planted stands from conditionally native populations both by three loci (Fe-2, Idh-1, Mdh-3) and by the totality of 18 polymorphic isoenzyme loci; the short-term population differs from conditionally native population only by two loci. The value of the inbreeding coefficient by the Idh-1 locus is significantly higher in both populations infested with the bark beetle than in “healthy” populations. The results of conducted studies demonstrate the necessity of continuation of the study on the gene pool state in Norway spruce populations owing to the degree of their infestation by the bark beetle along with the study on the dynamic state of the communities; this can provide a key to solving the problem of the forest preservation from pests.

Development of microsatellite genetic markers in Siberian stone pine ( Pinus sibirica Du Tour) based on the de novo whole genome sequencing

Siberian stone pine, Pinus sibirica Du Tour is one of the most economically and environmentally important forest-forming species of conifers in Russia. To study these forests a large number of highly polymorphic molecular genetic markers, such as microsatellite loci, are required. Prior to the new high-throughput next generation sequencing (NGS) methods, discovery of microsatellite loci and development of micro-satellite markers were very time consuming and laborious. The recently developed draft assembly of the Siberian stone pine genome, sequenced using the NGS methods, allowed us to identify a large number of microsatellite loci in the Siberian stone pine genome and to develop species-specific PCR primers for amplification and genotyping of 70 microsatellite loci. The primers were designed using contigs containing short simple sequence tandem repeats from the Siberian stone pine whole genome draft assembly. Based on the testing of primers for 70 microsatellite loci with tri-, tetra- or pentanucleotide repeats, 18 most promising, reliable and polymorphic loci were selected that can be used further as molecular genetic markers in population genetic studies of Siberian stone pine.

Spatial distribution of intron 2 of nad1 gene haplotypes in populations of Norway and Siberian spruce (Picea abies–P. obovata) species complex

The length and sequence variations among intron 2 haplotypes of the mitochondrial nad1 gene have been studied in the Norway and Siberian spruce (Picea abies (L.) H. Karst.–P. obovata Ledeb.) species complex. Twenty-two native populations and 15 provenances were analyzed. The distribution of the northern European haplogroup (haplotypes 721, 755, 789, 823, 857, 891, and 925) is delimited in the east by the Ural region inclusively. Haplotype 712 is widespread in populations of Siberia and in the Far East and in northeastern Russia. A novel variant of the Siberian haplogroup (780) containing three copies of the first minisatellite motif (34 bp) was found for the first time. The absence of an admixture of the northern European and Siberian haplotypes in the zone of spruce species introgression previously marked by morphological traits and nuclear allozyme loci was demonstrated. This may be evidence of the existence of a sharper geographic boundary between the two haplogroups, as compared to a boundary based on phenotypic and allozyme data. A high proportion of the interpopulation component of variation (65%) estimated by AMOVA indicates a substantial genetic subdivision of European and Siberian populations of the Palearctic spruce complex by mtDNA, which can be putatively explained by natural barriers to gene flow with seeds related, for instance, to the woodless regions of the western Siberian Plain in the Pleistocene and perhaps to the floodplains of large rivers.

Siberian Whitefish (Coregonus lavaretus pidschian, Coregonidae) from the Anabar River: Morphogenetic Structure of the Population

Data on the morphology, biology, and genetics of two ecological forms of pidschian-like whitefishes of the genus Coregonus, corresponding to the descriptions of East Siberian whitefish, C. lavaretus pidschian natio brachymystax, and glacial-plain whitefish, C. lavaretus pidschian natio glacialis, from the Anabar River of western Sakha (Yakutia) Republic (Russia), are presented. According to meristic characters, no differences are found between these ecological forms, while on the basis of plastic traits and occupied ecological niches these whitefishes differ substantially. We demonstrate that, on the basis of the sequences of the ND1 gene of the mtDNA, the East Siberian whitefishes are only slightly different from the whitefishes from the lower Ob River and Lake Teletskoye (Upper the Ob River basin), and probably have a common origin with them. Glacial-plain whitefishes from the Anabar River have a common origin with whitefishes from the water bodies of the Taimyr Peninsula and represent a complex of forms/species.

Taxonomic status and origin of some ecological forms of whitefish Coregonus lavaretus (L.) from water bodies of Siberia

Molecular genetic analysis of some ecological forms of whitefish belonging to the Coregonus lavaretus (L.) complex from different water bodies of Siberia revealed a high degree of differentiation between them. On the basis of the analysis of the mitochondrial DNA protein-coding ND1 gene, it was demonstrated that the previously described ecological forms of C. anaulorum, Yenisei River whitefish, or Issatschenko’s whitefish (C. fluviatilis), and cisco-like whitefish from Lake Baunt were good biological species. Moreover, each ecological form was represented by a number of phylogenetic lineages, one of which was species-specific. The remaining lineages were characterized by the mitochondrial DNA of another whitefish species, usually, the most common in each geographic region. The results of genetic analysis demonstrated that the formation of modern ecological forms of Siberian pidschian-like whitefish was accompanied by natural introgressive hybridization. It is suggested that most of the modern pidschian-like whitefish species originated as a result of reticulate evolution.

Molecular Identification and Karyological Analysis of a Rampant Aspen Populus tremula L. (Salicaceae) Clone

A rampant highly heterozygous aspen (Populus tremula L.) clone “Meshabash” has been revealed in course of population genetic diversity analysis in a native stand in the Republic of Tatarstan, Russia. Here we report the results of karyological analysis showing that this highly vigorous clone is diploid (2n = 38) while typically triploid aspen demonstrates increased growth rate and resistance to aspen trunk rot caused by fungus Phellinus tremulae. By means of DNA identification of a series of model trees using 14 SSR loci we outlined the area occupied by this clone (at least 1.94 ha) and demonstrated that its ramets constitute 40 out of 48 genotyped trunks on the plot with the maximal distance between ramets 254 m. Since aspen is able to regenerate after cutting or die-off of maternal tree by root suckers at a distance up to 20–35 m this assumed that current stand appeared as a result of such spreading from an ortet tree during at least 5 generations. Trunk rot damage in the wood of model trees indicated low influence of this pathogen on viability and performance of the studied clone that can be associated with its extreme heterozygosity level (0.926) exceeding all the studied trees in this research plot and in three other control samples.