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Since its introduction to North America in the early 1900s, white pine blister rust (WPBR) caused by the fungal pathogen has resulted in substantial economic losses and ecological damage to native North American five-needle pine species. The high susceptibility and mortality of these species, including limber pine ( ), creates an urgent need for the development and deployment of resistant germplasm to support recovery of impacted populations. Extensive screening for genetic resistance to WPBR has been underway for decades in some species but has only started recently in limber pine using seed families collected from wild parental trees in the USA and Canada. This study was conducted to characterize Alberta limber pine seed families for WPBR resistance and to develop reliable molecular tools for marker-assisted selection (MAS). Open-pollinated seed families were evaluated for host reaction following controlled infection using basidiospores. Phenotypic segregation for presence/absence of stem symptoms was observed in four seed families. The segregation ratios of these families were consistent with expression of major gene resistance (MGR) controlled by a dominant R locus. Based on linkage disequilibrium (LD)-based association mapping used to detect single nucleotide polymorphism (SNP) markers associated with MGR against , MGR in these seed families appears to be controlled by or other R genes in very close proximity to . These associated SNPs were located in genes involved in multiple molecular mechanisms potentially underlying limber pine MGR to , including NBS-LRR genes for recognition of effectors, signaling components, and a large set of defense-responsive genes with potential functions in plant effector-triggered immunity (ETI). Interactions of associated loci were identified for MGR selection in trees with complex genetic backgrounds. SNPs with tight Cr4-linkage were further converted to TaqMan assays to confirm their effectiveness as MAS tools. This work demonstrates the successful translation and deployment of molecular genetic knowledge into specific MAS tools that can be easily applied in a selection or breeding program to efficiently screen MGR against WPBR in Alberta limber pine populations.

Designer niches in which environmental variables are controlled are useful in forest restoration to enhance survival of planted tree seedlings. Here, we evaluate particular manipulated habitats, on site variables, and pre-seedling conditions hypothesized to improve the survival rate of whitebark pine (Pinus albicaulis) seedlings out-planted in Waterton Lakes National Park. The tree species is in peril due to blister rust and mountain pine beetle infestations in its range; and is a restoration priority in Waterton Lakes because populations in the park are highly infected with blister rust (up to 90%). At Summit Lake, 21 plots were set up and half of each was terra-torched; 1000 seedlings were planted in clusters of three, under four conditions: on burned areas in burned beargrass mats, in burned areas where beargrass mats were not present, in unburned areas where beargrass was present, and in unburned areas without beargrass. This study reports data for the seventh year after planting, and overall, survival was 53% for individual seedlings and at least one seedling survived in 60.8% of clusters. Planting in burned areas increased cluster survival (by 34.3%, p ˂ 0.0001) and planting near microsites increased cluster survival (by 19.3%, p ˂ 0.0001); the type of microsite did not make a difference. Planting in beargrass mats decreased survival, but not significantly (8.9%, p = 0.11) and this was true for burns, not unburned areas. Inoculation with native ectomycorrhizal fungi did not enhance survival most likely because controls on lightly terra-torched and unburned areas had access to local native fungi. This is the first study to report statistics on the planting of seedlings in clusters; the results need to be compared with studies where seedlings are planted individually.