Genomic prediction of resistance to Hymenoscyphus fraxineus in common ash (Fraxinus excelsior L.) populations.

The increase in introduced insect pests and pathogens due to anthropogenic environmental changes has become a major concern for tree species worldwide. Common ash (Fraxinus excelsior L.) is one of such species facing a significant threat from the invasive fungal pathogen Hymenoscyphus fraxineus. Some studies have indicated that the susceptibility of ash to the pathogen is genetically determined, providing some hope for accelerated breeding programs that are aimed at increasing the resistance of ash populations. To address this challenge, we used a genomic selection strategy to identify potential genetic markers that are associated with resistance to the pathogen causing ash dieback. Through genome-wide association studies (GWAS) of 300 common ash individuals from 30 populations across Poland (ddRAD, dataset A), we identified six significant SNP loci with a p-value ≤1 × 10-4 associated with health status. To further evaluate the effectiveness of GWAS markers in predicting health status, we considered two genomic prediction scenarios. Firstly, we conducted cross-validation on dataset A. Secondly, we trained markers on dataset A and tested them on dataset B, which involved whole-genome sequencing of 20 individuals from two populations. Genomic prediction analysis revealed that the top SNPs identified via GWAS exhibited notably higher prediction accuracies compared to randomly selected SNPs, particularly with a larger number of SNPs. Cross-validation analyses using dataset A showcased high genomic prediction accuracy, predicting tree health status with over 90% accuracy across the top SNP sets ranging from 500 to 10,000 SNPs from the GWAS datasets. However, no significant results emerged for health status when the model trained on dataset A was tested on dataset B. Our findings illuminate potential genetic markers associated with resistance to ash dieback, offering support for future breeding programs in Poland aimed at combating ash dieback and bolstering conservation efforts for this invaluable tree species.

Genetic resources of common ash (Fraxinus excelsior L.) in Poland.

BACKGROUND: Knowledge of genetic structure and the factors that shape it has an impact on forest management practices. European ash (Fraxinus excelsior L.) has declined dramatically throughout its range as a result of a disease caused by the fungus Hymenoscyphus fraxineus. Despite the need for conservation and restoration of the species, genetic data required to guide these efforts at the country level are scarce. Thereofore, we studied the chloroplast and nuclear genetic diversity of 26 natural common ash populations (1269 trees) in Poland. RESULTS: Chloroplast polymorphisms grouped the populations into two geographically structured phylogenetic lineages ascribed to different glacial refugia (the Balkans and the Eastern Alps). However, the populations demonstrated high genetic diversity (mean AR = 12.35; mean Ho = 0.769; mean He = 0.542) but low differentiation based on nuclear microsatellites (FST = 0.045). Significant spatial genetic structure, consistent with models of isolation by distance, was detected in 14 out of 23 populations. Estimated effective population size was moderate-to-high, with a harmonic mean of 57.5 individuals per population. CONCLUSIONS: Genetic diversity was not homogeneously distributed among populations within phylogenetic gene pools, indicating that ash populations are not equal as potential sources of reproductive material. Genetic differences among populations could be related to their histories, including founder effects or gene flow between evolutionary lineages (admixture). Our results suggest that ash stands across Poland could be treated as two main management units (seed zones). Therefore, despite the homogenizing effect of pollen gene flow known for this species, the genetic structure should be taken into account in the management of the genetic resources of the common ash. Although ash dieback poses an additional challenge for the management of genetic resources, efforts should be directed towards protecting populations with high genetic diversity within defined phylogenetic units, as they may be an important source of adaptive variation for future stands.

Population history, genetic variation, and conservation status of European white elm (Ulmus laevis Pall.) in Poland.

Key message: The core populations of the European white elm (Ulmus laevis Pall.) located in Poland maintained slightly higher level of genetic diversity compared to the peripheral populations of this species. Context: The most severe threat to elms is the loss of natural habitat under the pressures of agriculture and forestry as well as urbanization. The reductions in European white elm populations as well as populations of other elm species have also been caused by Dutch elm disease (DED). Previous studies have indicated a low level of genetic variation in Ulmus leavis Pall. However, in Poland, the genetic resources and demographic history of U. laevis populations remain poorly documented. Aims: The genetic resources of U. laevis in Poland were identified and characterized. Additionally, tests were performed to identify potential bottleneck signatures and effective population sizes of the examined populations. Methods: Polymorphism was analyzed using a set of six nuclear microsatellite markers (nSSRs) for 1672 individuals from 41 populations throughout the species range in Poland. Results: (1) A moderate level of genetic variation was found. (2) A low genetic differentiation and lack of population structuring were identified. (3) Evidence of reduction in population size was found as a consequence of severe, past bottlenecks. Conclusion: The loss of genetic diversity of U. laevis probably occurred in their refugia or shortly after the postglacial recolonization. This loss may have been affected by past DED pandemics similar to those seen at present.

Successful cryopreservation of Quercus robur plumules.

Successful cryopreservation of Q. robur germplasm as plumules (i.e. shoot apical meristems of embryos) is described in this paper. After excision from the recalcitrant seeds and preliminary storage in 0.5 M sucrose solution (18 h), the plumules were subjected to cryoprotection (in 0.75 M sucrose, followed by 1.0 M sucrose and 1.5 M glycerol solutions), and next to desiccation (over silica gel or in nitrogen gas) and cooling (in slush at -210°C or in vials filled with liquid nitrogen, LN, -196°C), and were then cryostored for 24 h. High percentage of survival was obtained after cryostorage (21-67%, depending on pretreatment, assessed in vitro by greening plumules that increased in size). Desiccation of plumules over silica gel resulted in significantly higher survival after cryopreservation (58%) in comparison with desiccation in nitrogen gas (29%), with regrowth (shoots with leaves) 5-18%. The extent of plumule desiccation was comparable in both methods, in which drying of plumules for 20 min decreased the water content to 0.5-0.6 g H(2)O g(-1) dry weight before LN exposure. The type of LN exposure did not significantly influence plumule survival and regrowth after cryostorage. Plumules isolated from acorns of four provenances survived cryostorage after cryoprotection followed by desiccation over silica gel and direct cooling in vials with LN (survival 51-76%, regrowth 8-20%). Normal plants developed from the recovered shoots after rooting. The presented protocol for Q. robur plumule cryopreservation may offer a potential approach for establishing germplasm conservation in gene banks for Quercus species.