Yana Sboeva

The Global warming processes and the large-scale impact of invasive species, such as the Ussuriysk polygraph, lead to the degradation of forest ecosystems and their biological diversity. The purpose of the research was to identify the genetic factors of the resistance of coniferous plant species to drought using the example of Scots pine (Pinus sylvestris L.) and a hybrid complex of Norway and Siberian spruce (Picea abies (L.) H. Karst.–P. obovata Ledeb.) and to analyze the genetic determinants of the resistance of coniferous plant species to pathogens and phytophages in the Ural region of the Ussuriysky polygraph invasion and in the adjacent territories. 16 pairs of primers for the loci of adaptively significant genes of P. sylvestris and P. abies–P. obovata have been developed and tested. P. sylvestrisassociated with drought resistance; 97 and 110 polymorphic positions in the studied loci of the species, respectively, were identified. It was found that the studied species have an average level of nucleotide diversity in the region for both P. sylvestris (Hd = 0,662; π = 0,004; θW = 0,013) and P. abies–P. obovata (Hd= 0,761; π = 0,005; θW = 0,008). Loci have been identified whose polymorphism is potentially adaptive in relation to abiotic (drought resistance) and biotic (impact of pathogenic organisms) environmental factors, which are promising for further study. The polymorphism of the loci of adaptively significant genes associated with resistance to drought and the effects of pathogenic organisms and phytophages is recommended to be taken into account when assessing the state of the gene pools of populations of coniferous plant species in the research region.

Background/Objectives: The spruces of the Picea abies–P. obovata complex have a total range that is the most extensive in the world flora of woody conifers. Hybridization between the nominative species has led to the formation of a wide introgression zone, which probably increases the adaptive potential of the entire species complex. This study aimed to search the genes associated with drought resistance, develop primers for the informative loci of these genes, identify and analyze SNPs, and establish the parameters of nucleotide diversity in the studied populations. Methods: The objects of this study were eight natural populations of the spruce complex in the Urals. Nucleotide sequences related to drought resistance spruce genes with pronounced single-nucleotide substitutions were selected, based on which 16 pairs of primers to their loci were developed and tested. Results: Based on the developed primers, six pairs of primers were chosen to identify SNPs and assess the nucleotide diversity of the studied populations. All selected loci were highly polymorphic (6 to 27 SNPs per locus). It was found that the Pic01 locus is the most variable (Hd = 0.947; π = 0.011) and selectively neutral, and the Pic06 locus is the most conservative (Hd = 0.516; π = 0.002) and has the most significant adaptive value. Conclusions: The nucleotide diversity data for the studied populations reveal similar values among the populations and are consistent with the literature data. The discovered SNPs can be used to identify adaptive genetic changes in spruce populations, which is essential for predicting the effects of climate change.

Background: Scots Pine is one of the main forest-forming species in boreal forests; it has great economic and ecological significance. This study aimed to develop and test primers for detecting nucleotide polymorphisms in genes that are promising for detecting adaptive genetic variability in populations of Pinus sylvestris in the Urals and adjacent territories. Objectives: The objects of the study were 13 populations of Scots Pine located in the Perm Territory, Chelyabinsk Region, and the Republic of Bashkortostan. Results: Sixteen pairs of primers to loci of potentially adaptively significant genes were developed, from which three pairs of primers were selected to detect the nucleotide diversity of the studied populations. The indicator of total haplotype diversity determined in the three studied loci varied from 0.620 (Pinus-12 locus) to 0.737 (Pinus-11 locus) and, on average, amounted to 0.662. The nucleotide diversity indicators in P. sylvestris in the study region were, on average, low (π = 0.004, θ_W = 0.013). Their highest values were found at the Pinus-12 locus (π = 0.005; θ_W = 0.032), and the lowest were found at the Pinus-15 locus (π = 0.003; θ_W = 0.002). This indicates that Pinus-15 is the most conserved of the three studied loci. In the three studied P. sylvestris loci associated with adaptation to environmental factors, 97 polymorphic positions were identified. The 13 populations of P. sylvestris are characterized by an average level of genetic diversity (Hd = 0.662; π = 0.004; θ = 0.013). Conclusions: The polymorphic loci of adaptively significant genes of P. sylvestris can help identify the adaptive potential of pine forests in conditions of increasing ambient temperatures.

Genetic diversity is important for the long-term survival of species and plays a critical role in their conservation. To manifest the adaptive potential, it is necessary to preserve the allelic diversity of populations, including both typical and region-specific alleles. Molecular genetic analysis of 22 populations of Scotch pine (Pinus sylvestris L.; Pinaceae) in 10 subjects of the Russian Federation in the East European Plain and the Middle Urals was carried out. Molecular genetic analysis of 22 populations of P. sylvestris revealed 182 polymorphic PCR fragments. The studied populations are characterized by a medium level of genetic diversity. A high subdivision coefficient (G_ST) of the studied populations was established; the intensity was 0.559. At the same time, the level of subdivision differed for different regions; for the populations from the Middle Urals, it was 15.5% (G_ST = 0.155), and for the populations from the East European Plain, it was 55.8% (G_ST = 0.558). The dendrogram of genetic similarity shows five clusters of the studied populations of P. sylvestris according to their geographical location. The populations from the East European Plain are mostly characterized by typicality, while the populations from the Middle Urals, on the contrary, are more specific in gene pools. The use of the coefficient of genetic originality to identify populations with typical and specific alleles allows for solving the problem of selecting populations for the conservation of forest genetic resources. The data obtained on genetic diversity, and the structure of populations growing in areas of active logging, are important for determining the geographical origin of plant samples, which is an integral part of the control of illegal logging.

74 ISSR-PCR markers were isolated in four studied populations of Scots pine in the Perm Territory, of which 67 (P95=0.905) were polymorphic. The studied populations are characterized by high rates of genetic diversity (P95=0.905; HE=0.187; ne=1.402; R=12). It was found that among the studied 4 populations of P. sylvestris, the population from the Gainskiy forestry (P95=0.842; HE=0.212; ne=1.358; R=1) possesses the highest genetic diversity, and the population from the Perm forestry (P95=0.800; HE=0.173; ne=1.282; R=1). A comparative analysis of the literature and the data obtained showed that the indicators of the genetic diversity of the studied populations of Scots pine in the Perm region have average values for the species (P95=0.905; HE=0.187; na=1.905; ne=1.402).

Based on the analysis of the polymorphism of two types of microsatellite markers (ISSR and SSR), the state of gene pools of the fifteen of Siberian larch populations from three regions of the Urals: Northern, Middle and Southern was estimated. The parameters of genetic diversity were revealed, its structure was established at the intrapopulation level. To assess the uniqueness of the gene pool, we used the coefficient of genetic originality (KGO), the analysis of which revealed populations with typical and region-specific gene pools. It was established that the studied samples are generally characterized by a high level of genetic diversity. It was found that the gene pools of samples from the South Urals are characterized by the greatest specificity, the lowest values of KGO, i. e. more typical gene pools, are noted in the North Urals samples of L. sibirica, the average values of KGO are from the samples of the Middle Urals. Also, 3 unique alleles were found in the ZIL, BND, and KCH samples, in the rest, no unique markers were detected. For a comprehensive assessment of the state of the gene pools of populations, all established indicators of genetic diversity have been transferred to the scale for assessing the status of gene pools developed on the example of the studied L. sibirica populations. Based on data on genetic diversity obtained using two types of molecular markers, it was found that the gene pools of ten studied L. sibirica populations are in satisfactory condition, and five have signs of gene pool degradation. Based on the results of the study, recommendations are made on the conservation of L. sibirica genetic resources in the Urals.

A study of the genetic structure and differentiation of eight populations of Pinus sylvestris L. on the Russian Plain showed that the populations of PsI and PsII (D=0.066) are located at the smallest genetic distance, and between the populations of PsI and PsIV (D=0.308) at the greatest genetic distance. On the dendrogram, the studied populations formed four clusters: PsI and PsII; PsIII and PsIV; PsV and PsVI; PsVII and PsVIII. Analysis of the genetic structure of eight populations of P. sylvestris showed that the expected proportion of heterozygous genotypes (HT) for the total sample was 0.320, the expected proportion of heterozygous genotypes in a single population for all loci (HS) was 0.170, therefore, the population subdivision (GST) was high and amounted to 0.468. The studied populations are highly differentiated, since the interpopulation component accounts for 46.8% of the genetic diversity. In all studied populations, the indicator h has values less than 0.3. An analysis of the fraction of rare alleles showed that the genetic structure is less balanced in the populations PsIII (h=0.254) and PsIV (h=0.273). The most balanced genetic structure in the populations of PsVII (h=0.112) and PsVIII (h=0.127). Data on the genetic structure and differentiation of the common pine populations should be taken into account when developing recommendations for preserving their gene pools.