Quantitative Disease Resistance to White Pine Blister Rust at Southwestern White Pine’s (Pinus strobiformis) Northern Range

White pine blister rust, caused by the non-native, invasive fungal pathogen Cronartium ribicola, is a significant cause of mortality in white pines (Pinus subgenus Strobus) in North America. Along with climate-driven range contraction, mortality from blister rust can seriously impact the abundance and distribution of the nine white pine species native to the United States and Canada. Very little evaluation of this disease in southwestern white pine (Pinus strobiformis) has been previously undertaken, but genetic resistance to the disease has been documented, including major gene resistance (MGR) conferred by a dominant R gene. Data is emerging suggesting that the species also has quantitative disease resistance (QR). Our results suggest QR occurs at low frequency, with perhaps 10% of trees having a moderate level (> 35% survival). We assessed progeny arrays from 40 P. strobiformis families (1873 seedlings), originating from three populations, inoculated with C. ribicola. Subsequently, the seedlings were assessed for signs, symptoms and resulting impact in a common garden trial over a 7.5-year period to determine the types and frequency of resistance in a portion of this species’ range. There was a high incidence of both stem symptoms and mortality in the P. strobiformis families tested, and families ranged in survival from 0 to 84.6%. Three families had > 70% survival, representing perhaps the highest documented QR to date in a North American white pine species. Approximately 29.1% of the 441 surviving seedlings showed no stem symptoms, and of the approximately 70.8% of seedlings surviving with infections only few (24 of 316) had infections of moderate to high severity. QR traits associated with improved survival were primarily related to lower severity of infection, a reduced number of stem symptoms, and an increased number of bark reactions. Despite the high overall susceptibility, the presence of QR appears to be at a frequency and level useful to forest managers involved in restoration and reforestation efforts.

Comparative association mapping reveals conservation of major gene resistance to white pine blister rust in southwestern white pine (Pinus strobiformis) and limber pine (P. flexilis)

All native North American white pines are highly susceptible to white pine blister rust (WPBR) caused by Cronartium ribicola. Understanding genomic diversity and molecular mechanisms underlying genetic resistance to WPBR remains one of the great challenges in improvement of white pines. To compare major gene resistance (MGR) present in two species, southwestern white pine (Pinus strobiformis) Cr3 and limber pine (P. flexilis) Cr4, we performed association analyses of Cr3-controlled resistant traits using SNP assays designed with Cr4-linked polymorphic genes. We found that ~ 70% of P. flexilis SNPs were transferable to P. strobiformis. Furthermore, several Cr4-linked SNPs were significantly associated with the Cr3-controlled traits in P. strobiformis families. The most significantly associated SNP (M326511_1126R) almost co-localized with Cr4 on the Pinus consensus linkage group 8 (LG-8), suggesting that Cr3 and Cr4 might be the same R locus, or have localizations very close to each other in the syntenic region of the P. strobiformis and P. flexilis genomes. M326511_1126R was identified as a non-synonymous SNP, causing amino acid change (Val376Ile) in a putative pectin acetylesterase (PAE), with coding sequences identical between the two species. Moreover, top Cr3-associated SNPs were further developed as TaqMan genotyping assays, suggesting their usefulness as marker-assisted selection (MAS) tools to distinguish genotypes between quantitative resistance (QR) and MGR. This work demonstrates the successful transferability of SNP markers between two closely related white pine species in the hybrid zone, and the possibility for deployment of MAS tools to facilitate long-term WPBR management in P. strobiformis breeding and conservation.

Verbenone and Green-Leaf Volatiles Reduce Whitebark Pine Mortality in a Northern Range-Expanding Mountain Pine Beetle Outbreak

This is the first study testing effectiveness of semiochemical treatments to protect individual trees from a range-expanding mountain pine beetle (MPB, Dendroctonus ponderosae Hopkins) attack into newly exposed host populations of endangered whitebark pine (Pinus albicaulis Engelmann). We investigated the effectiveness of a combination of verbenone and Green-Leaf Volatiles (GLV) to protect rare and valuable disease-resistant trees during a MPB epidemic from 2015 to 2018 in Jasper National Park, Canada. Treatments reduced the proportion of trees attacked by MPB for all diameter classes, across all stands, from 46 to 60%. We also evaluated the effect of the exotic disease white pine blister rust (caused by the fungus Cronartium ribicola J.C. Fisch), the species’ other main regional threat. MPB were less likely to attack large, rust infected trees than healthy trees, emphasizing the value of the semiochemical treatment. Protecting large, cone-bearing disease-resistant whitebark pine trees is fundamental to whitebark pine recovery. Maintaining reproductive trees on the landscape increases the frequency and diversity of rust-resistant genotypes more effectively than just planting seedlings to replace MPB-killed trees, because this slow-growing species takes over 80 years to reproduce. Our study confirmed protecting large rust-resistant trees with verbenone and GLV is a proactive and effective treatment against MPB for whitebark pine in naïve populations.

Using Hyperspectral Imagery to Detect an Invasive Fungal Pathogen and Symptom Severity in Pinus strobiformis Seedlings of Different Genotypes

Finding trees that are resistant to pathogens is key in preparing for current and future disease threats such as the invasive white pine blister rust. In this study, we analyzed the potential of using hyperspectral imaging to find and diagnose the degree of infection of the non-native white pine blister rust in southwestern white pine seedlings from different seed-source families. A support vector machine was able to automatically detect infection with a classification accuracy of 87% (κ = 0.75) over 16 image collection dates. Hyperspectral imaging only missed 4% of infected seedlings that were impacted in terms of vigor according to expert’s assessments. Classification accuracy per family was highly correlated with mortality rate within a family. Moreover, classifying seedlings into a ‘growth vigor’ grouping used to identify the degree of impact of the disease was possible with 79.7% (κ = 0.69) accuracy. We ranked hyperspectral features for their importance in both classification tasks using the following features: 84 vegetation indices, simple ratios, normalized difference indices, and first derivatives. The most informative features were identified using a ‘new search algorithm’ that combines both the p-value of a 2-sample t-test and the Bhattacharyya distance. We ranked the normalized photochemical reflectance index (PRIn) first for infection detection. This index also had the highest classification accuracy (83.6%). Indices such as PRIn use only a small subset of the reflectance bands. This could be used for future developments of less expensive and more data-parsimonious multispectral cameras.