Pinus cembra samples were from a previous comparative study (European larch and Swiss stone pine) (Jahn and Heinze, unpublished), for which material was sampled from six sites along the Austrian Alps (Table S1), 15 individuals from each population. sylvestris, and (3) genotype by PCR and Sanger sequencing Scots pine samples of different habitat types by revealing possible nucleotide variation at these loci. cembra transcriptome to formerly annotated stress responsive genes, (2) test on the applicability of primers designed on these gene fragments in P. The study was performed to describe (1) homologues in the P. Evaluating genetic variation of the species with focus on the stress-adaptive genes by appropriate genetic tools could provide useful information for the conservation of native remnant populations as a biodiversity resource for the future. Scots pine, as a widely tolerant pioneer species, surviving close to the permafrost during the Pleistocene has adapted to different climates, being able to colonize even recently man-disturbed areas. Haploxylon Swiss stone pine ( Pinus cembra L.), a soft pine, which has one fibrovascular bundle and diploxylon Scots pine ( Pinus sylvestris L.), a hard pine, which has two, both belonging to subgenus Pinus but belonging to different subsections namely Strobus and Pinus and the latter tending to have harder timber and a larger amount of resin, have formerly been described as highly sensitive to climatic changes. As an alternative, massively parallel transcriptome sequencing is an efficient route to develop genetic markers, which can be used in genetic analysis of different conifer species. This is an obstacle to the genetic evaluation of a large number of species. Nevertheless, in case of conifers, due to the large genomes, only a few species have been sequenced, so far. Īn important way to obtain genetic information is by genome sequencing. In these conditions, taking into consideration the genetic information related to adaptation is fundamental in developing conservation strategies. Aridity influences drought-response trait differentiation and genetic divergence among populations. The SNPs detected may have important applications in further studies of adaptive genetic variation, providing tools to study relevant genes important in the long-term adaptation of pine species.įocus on plant response to environmental stress is becoming increasingly important, as most future climate scenarios suggest climate change that imply an increase in aridity of many areas, causing abiotic stress and seriously threatening natural ecosystems. cembra and tested for transferability in Scots pine, our results give support for the use of de novo markers targeting conserved regions across different pines. By providing de novo molecular markers developed in P. In a total of 3735 sites no indels, eight synonymous and 11 non-synonymous SNPs were found. After sequencing and BLAST search of the amplified products, parts of 19 different candidate genes were analysed by considering the polymorphic sites, insertions/deletions as well as synonymous and non-synonymous SNPs. sylvestris DNA samples originating from three different types of habitat. In this study novel PCR markers, based on stress responsive genes, were designed from the transcriptome of the haploxylon Swiss stone pine ( Pinus cembra L.) and tested for SNPs in the diploxylon Scots pine ( Pinus sylvestris L.). The analysis of their sequence helps in developing molecular markers, such as SNPs, which represent a useful tool in detecting adaptive signals in populations. Massively parallel sequencing of cDNA is an efficient route for generating sequence collections that represent expressed genes under different environmental control.

Codoncode aligner ver. 2