Development of molecular genetic markers of genes encoding adaptive response methods in pine

Under the conditions of global climate change, which leads to increased frequency of droughts, frosts, and pest outbreaks, the breeding of Scots pine (Pinus sylvestris L.) for enhanced resilience is an urgent task. Marker-assisted selection (MAS) is a promising tool for accelerating the breeding process, requiring the development of molecular markers associated with adaptive traits. The aim of this study was to develop and preliminarily test such markers. Due to the lack of a whole-genome reference assembly for Scots pine, a candidate-gene approach was employed. The study focused on genes whose homologs in related species are associated with adaptation: PRR1 and PRR7 (components of the circadian clock involved in response to photoperiod and temperature stress), dhn1 (a dehydrin conferring resistance to dehydration), as well as the mitochondrial genes COX1 and ATP-A (phylogenetic markers). Based on polymorphisms (SNPs and a deletion) identified by sequencing, 4 KASP and 1 STS markers were developed. Their testing was conducted on geographical cultures of pine using a sample of 100 trees from four climotypes (Leningrad Region, Karelia, Estonia, Udmurtia). Statistical analysis using Fisher's exact test revealed a significant difference (p ≤ 0.05) in allele frequencies of the PRR7 gene between the Karelian and Udmurtian climotypes, indicating a potential longitudinal cline for this locus and its potential informativeness for assessing adaptive population differentiation. For the other genes, no clear associations were established within this sample. The results demonstrate the effectiveness of the candidate-gene approach for the initial stage of developing molecular markers for adaptation in Scots pine and lay the groundwork for further research to establish reliable phenotype-genotype linkages with the aim of subsequent application in breeding programs to create resilient planting material.

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