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.

How do pathogens evolve novel virulence activities?

We consider the state of knowledge on pathogen evolution of novel virulence activities, broadly defined as anything that increases pathogen fitness with the consequence of causing disease in either the qualitative or quantitative senses, including adaptation of pathogens to host immunity and physiology, host species, genotypes, or tissues, or the environment. The evolution of novel virulence activities as an adaptive trait is based on the selection exerted by hosts on variants that have been generated de novo or arrived from elsewhere. In addition, the biotic and abiotic environment a pathogen experiences beyond the host may influence pathogen virulence activities. We consider pathogen evolution in the context of host-pathogen evolution, host range expansion, and external factors that can mediate pathogen evolution. We then discuss the mechanisms by which pathogens generate and recombine the genetic variation that leads to novel virulence activities, including DNA point mutation, transposable element activity, gene duplication and neofunctionalization, and genetic exchange. In summary, if there is an (epi)genetic mechanism that can create variation in the genome, it will be used by pathogens to evolve virulence factors. Our knowledge of virulence evolution has been biased by pathogen evolution in response to major gene resistance, leaving other virulence activities underexplored. Understanding the key driving forces that give rise to novel virulence activities, and the integration of evolutionary concepts and methods with mechanistic research on plant–microbe interactions, can help inform crop protection.

Root Rot of Juniperus and Microbiota by Phytophthora lateralis in Oregon Horticultural Nurseries

Widespread symptoms of root rot and mortality on Juniperus communis and Microbiota decussata were observed in two horticultural nurseries in Oregon, leading to the isolation of a Phytophthora sp. from diseased roots. Based on morphology and sequencing the internal transcribed spacer ITS1-5.8S-ITS2 region, isolates were identified as the invasive pathogen Phytophthora lateralis, causal agent of Port-Orford-cedar (POC; Chamaecyparis lawsoniana) root disease. Additional sequencing of the cytochrome c oxidase subunit 1 and 2 genes identified all isolates as belonging to the PNW lineage. Utilizing recovered isolates plus a POC-wildlands isolate and susceptible POC as controls, we completed Koch’s postulates on potted Juniperus and Microbiota plants. Nursery isolates were more aggressive than the forest isolate, which was used in the POC resistance breeding program. Increased aggressiveness was confirmed using a branch stem dip assay with four POC clones that differed in resistance, although no isolate completely overcame major-gene resistance. Isolates were sensitive to mefenoxam, a fungicide commonly used to suppress Phytophthora spp. growth in commercial nurseries. Although POC resistance is durable against these more aggressive nursery isolates, the expanded host range of P. lateralis challenges POC conservation through the continued movement of P. lateralis by the nursery industry.

Pyramiding Quantitative Resistance with a Major Resistance Gene in Apple: From Ephemeral to Enduring Effectiveness in Controlling Scab

Genetic resistance is a useful strategy to control plant disease, but its effectiveness may be reduced over time due to the emergence of pathogens able to circumvent the defenses of the plant. However, the pyramiding of different resistance factors in the same plant can improve the effectiveness and durability of the resistance. To investigate the potential for this approach in apple to control scab disease we surveyed scab incidence in two experimental orchards located at a distance of more than 300 km planted with apple genotypes carrying quantitative resistance and major gene resistance alone or in combination. Our results showed that the effectiveness of pyramiding in controlling scab was dependent on the site and could not be completely explained by the effectiveness level of the resistances alone.

Genetic resistance to white pine blister rust in limber pine (Pinus flexilis): major gene resistance in a northern population

Limber pine, Pinus flexilis E. James, a wide-ranging tree species in western North America, is highly susceptible to white pine blister rust (WPBR), caused by the non-native fungal pathogen Cronartium ribicola J.C. Fisch. The Canadian populations in particular have been heavily impacted, and in 2014, limber pine was designated endangered in Canada by the Committee on the Status of Endangered Wildlife in Canada (COSEWIC). Little is known about genetic resistance to WPBR in limber pine, but major gene resistance (MGR) has been characterized in some populations in the United States. This study examines resistance in seedling families from 13 parent trees from British Columbia, Alberta, and Oregon, representing the northern- and northwestern-most populations. Most families were susceptible, with 100% of the seedlings cankered, but one family from Alberta segregated 1:1 for cankered and canker free. This is the first report of (a) MGR in Canada of any of the four species of five-needle pines native to Canada and (b) any resistance in limber pine in Canadian populations and is the northernmost known incidence of putative R-gene resistance in a natural stand of any five-needle pine species. Many of the Canadian selections were from stands with high incidence of WPBR infection, and their high susceptibility in this trial suggests that further infection and mortality is likely in the Canadian populations.

Marker-Assisted Selection for Disease Resistance in Wheat and Barley Breeding

Marker-assisted selection (MAS) provides opportunities for enhancing the response from selection because molecular markers can be applied at the seedling stage, with high precision and reductions in cost. About each of 50 genes conferring monogenic resistances and hundreds of quantitative trait loci (QTL) for quantitative disease resistances have been reported in wheat and barley. For detecting single-major gene resistance, MAS could be easily applied, but is often not necessary because the resistances are selected phenotypically. In quantitative disease resistances, MAS would be very useful, but the individual QTL often have small effects. Additionally, only a few monogenic resistances are durable and only a few QTL with high effects have been successfully transferred into elite breeding material. Further economic and biological constraints, e.g., a low return of investment in small-grain cereal breeding, lack of diagnostic markers, and the prevalence of QTL–background effects, hinder the broad implementation of MAS. Examples in which MAS has been successfully applied to practical breeding are the wheat rust resistance genes Lr34 and Yr36, the eyespot resistance gene Pch1, the recessive resistance genes rym4/rym5 to barley yellow mosaic viruses, mlo to barley powdery mildew, and two QTL for resistance to Fusarium head blight in wheat (Fhb1 and Qfhs.ifa-5A). Newly identified broad-spectrum resistance genes/QTL conferring resistance to multiple taxa of pathogens offer additional perspectives for MAS. In the future, chip-based, high-throughput genotyping platforms and the introduction of genomic selection will reduce the current problems of integrating MAS in practical breeding programs and open new avenues for a molecular-based resistance breeding.

Can We Enhance Durable Resistance Against Plasmopara Halstedii (Sunflower Downy Mildew)?

Can We Enhance Durable Resistance AgainstPlasmopara Halstedii(Sunflower Downy Mildew)?Sunflower downy mildew caused byPlasmopara halstediiis one of the most potentially important diseases. So far, a complete, major gene resistance (Pl) has been used successfully, but with the appearance of eight races in France since 2000, research on more durable resistance was undertaken. In this study, we present new results concerning the evolution of pathogenicity ofP. halstediiunder conditions of re-enforced infection and differentPlgene selection pressures. Moreover, we imagine the evolution of virulence and aggressiveness ofP. halstediiunder a mixture model of sunflower inbred lines carrying the two types of resistance which may enhance durable resistance against it. Examples of host-parasite interactions including the influence of plant mixture models against pathogens are equally presented to understand how the pathogen develops its pathogenicicity.

Influence of Host Resistance on the Genetic Structure of the White Pine Blister Rust Fungus in the Western United States

Cronartium ribicola, the causal agent of white pine blister rust, has been devastating to five-needled white pines in North America since its introduction nearly a century ago. However, dynamic and complex interactions occur among C. ribicola, five-needled white pines, and the environment. To examine potential evolutionary influences on genetic structure and diversity of C. ribicola in western United States, population genetic analyses of C. ribicola were conducted using amplified fragment length polymorphism (AFLP) molecular markers. The fungus was sampled at six sites. Collections for two of the six sites were from separate plantings of resistant-selected western white pine and sugar pine. Heterozygosity based on polymorphic loci among populations ranged from 0.28 to 0.40, with resistant-selected plantations at the extremes. Genetic differentiation was also highest between these two populations. Principal coordinates analysis and Bayesian assignment placed most isolates that are putative carriers of virulence to major-gene resistance into a discernable cluster, while other isolates showed no clustering by site or host species. These results indicate that C. ribicola in western North America is not genetically uniform, despite its presumed single site of introduction and relatively brief residence. Moreover, major-gene resistance appears to have imposed strong selection on the rust, resulting in reduced genetic diversity. In contrast, no evidence of selection was observed in C. ribicola from hosts that exhibit only multigenic resistance.