Key message: Six new PCR-based markers for thePc39crown rust resistance gene inAvena sativaL. were developed.Pc39was mapped to Mrg11 of the oat consensus map using BLASTn analysis. The aim of this study was the identification of molecular markers for thePc39gene in cultivated oat (Avena sativaL.).Pc39is a major race-specific crown rust resistance gene originally found in an Israeli accession of the wild hexaploidAvena sterilis. The effectiveness of this gene in Europe has decreased in recent years, but is still relatively high and breeding programs would benefit from the availability of molecular markers to aid in its mapping and deployment. The complexity of the oat genome poses a significant obstacle to genetic research. No oat rust resistance genes have yet been cloned, and even the number of relevant molecular markers is very limited. Here, genotyping of a segregating population derived from a cross ‘Celer’ (Pc39)/STH9210 (susceptible) was conducted using RAPD- and SRAP-PCR-based methods, as well as microarray-based DArT™ and next-generation sequencing DArTseq™ techniques. Markers associated withPc39were placed on the hexaploid oat consensus linkage group Mrg11 at 3.7–6.7 cM. Six new PCR-based markers were developed to allow identification of the resistantPc39allele. These tightly linked markers will be useful in marker-assisted selection, with the closest, SCAR_3456624, being within 0.37 cM ofPc39. The newly developed markers could find applications in the fine mapping or positional cloning of this gene. Moreover, easy-to-use PCR-based markers linked toPc39could facilitate the utilization of this gene in oat breeding programs, especially as a component of crown rust resistance gene pyramids.

The crown rust fungus Puccinia coronata f. sp. avenae P. Syd. & Syd. (Pca) attacks cultivated oat and its wild relatives, causing significant losses to the crop worldwide. Although understanding the origin and dynamics of the pathogen’s diversity is critical to developing methods for its control, there are little relevant data on Pca virulence diversity in Europe, the global center of oat production. The goal of this study was to analyze the diversity of Pca populations in Poland in 2013 to 2015 based on their ability to overcome currently available host resistance Pc genes. Pca isolate virulence was evaluated on a panel of lines containing 26 major resistance genes of oat. The isolates were able to overcome from 1 to 16 resistance genes each, with most isolates being virulent on five to seven lines. In all years, a very high level of crown rust pathotype diversity was observed, with Simpson and Evenness indices of 0.99. In total, 156 different pathotypes were detected, with no prevalent pathotype in any of the 3 years analyzed. The results showed that the virulence level of P. coronata isolates was relatively low for each year studied (21% on average), most likely owing to the low take up of Pc genes in Polish oat cultivars, meaning that many sources of resistance are still effective against Pca races occurring in Poland. The long-range dispersal of Puccinia spores supported by the availability of wild, weedy, and cultivated Avena species makes it likely that the virulence profile seen in Poland is representative of much of central Europe and beyond.

Wild oat tetraploids of the section Pachycarpa have already been proven to be a rich source of useful genes but have largely been unexploited for Puccinia coronata resistance. In this study, accessions of Avena magna, A. murphyi, and A. insularis gathered from European and North American gene banks were evaluated at the seedling stage for crown rust reaction using the host–pathogen test and six highly diverse and virulent P. coronata isolates. Of the 92 Avena accessions analyzed, 58.7% were resistant to at least one crown rust race. In all, 37% of the tested accessions reacted nonuniformly, which indicated their heterogeneity. The highest level of resistance was observed in three of the accessions, one of which was verified by flow cytometry as being hexaploid and two of which were verified as being tetraploids. The infection profiles of 19 accessions corresponded to resistance determined by the genes Pc14, Pc39, Pc40, Pc48, Pc50, Pc54, Pc55, Pc61, Pc67, Pc68, Pc97, Pc101, or Pc104. The patterns of infection of the remaining resistant A. magna and A. murphyi accessions allowed us to postulate the presence of potentially novel crown rust resistance genes.

The aim of the present study was to identify Avena sterilis genotypes demonstrating a high level of resistance against oat powdery mildew, using host-pathogen tests. The study was conducted on 350 A. sterilis genotypes from different parts of the world. Six single-spore isolates of Blumeria graminis (DC.) f. sp. avenae, which demonstrated different levels of virulence to control lines and cultivars, were used in host-pathogen screening tests. To confirm the resistant response of selected genotypes, 13 other isolates were used. Reactions to the isolates were grouped into three classes: resistant, intermediate, and susceptible. Susceptible cultivars Sam and Fuchs were used as controls to estimate the degree of infection. The results of the screening test showed that 10 genotypes were classified as resistant. The second test based on 13 other isolates revealed that only four of the 10 genotypes were a valuable source of resistance against powdery mildew. The identified genotypes may be used in oat breeding programs to increase the level of resistance against powdery mildew. First, however, further studies aimed at identifying whether this resistance is conditioned by a single gene or combinations of different genes are required.

Crown rust, caused by Puccinia coronata f. sp. avenae, is one of the most destructive fungal diseases of oat worldwide. Growing disease-resistant oat cultivars is the preferred method of preventing the spread of rust and potential epidemics. The object of the study was Pc50-5, a race-specific seedling crown rust resistant gene, highly effective at all growth stages, selected from the differential line Pc50 (Avena sterilis L. CW 486-1 × Pendek). A comparison of crown rust reaction as well as an allelism test showed the distinctiveness of Pc50-5, whereas the proportions of phenotypes in segregating populations derived from a cross with two crown rust-susceptible Polish oat cultivars, Kasztan × Pc50-5 and Bingo × Pc50-5, confirmed monogenic inheritance of the gene, indicating its usefulness in oat breeding programs. Effective gene introgression depends on reliable gene identification in the early stages of plant development; thus, the aim of the study was to develop molecular markers that are tightly linked to Pc50-5. Segregating populations of Kasztan × Pc50-5 were genotyped using DArTseq technology based on next-generation Illumina short-read sequencing. Markers associated with Pc50-5 were located on chromosome 6A of the current version of the oat reference genome (Avena sativa OT3098 v2, PepsiCo) in the region between 434,234,214 and 440,149,046 bp and subsequently converted to PCR-based SCAR (sequence-characterized amplified region) markers. Furthermore, 5426978_SCAR and 24031809_SCAR co-segregated with the Pc50-5 resistance allele and were mapped to the partial linkage group at 0.6 and 4.0 cM, respectively. The co-dominant 58163643_SCAR marker was the best diagnostic and it was located closest to Pc50-5 at 0.1 cM. The newly discovered, very strong monogenic crown rust resistance may be useful for oat improvement. DArTseq sequences converted into specific PCR markers will be a valuable tool for marker-assisted selection in breeding programs.

Crown rust is the most widespread and damaging disease of oat (Avena species). Genetic resistance to the pathogen is the preferred method for crop protection but widespread deployment of limited numbers of major effect genes has promoted the rapid emergence and spread of pathogen races that are able to overcome these genes. Combining genes with even partial resistance may help develop durable cultivars that are less vulnerable to changes in pathogen virulence. Partial resistance is expected to be relatively common in populations of wild species where constant pathogen pressure encourages diversity in host resistance mechanisms, but it may be discarded in conventional screens for major gene resistance. Here, we used a detached leaf assay to detect resistance to the crown rust pathogen, Puccinia coronata Cda. f. sp. avenae, in previously uncharacterized collections of the hexaploid wild oat relative A. sterilis made by the Polish National Centre for Plant Genetic Resources. Many of the accessions were collected in Morocco, the center of diversity for the Avena genus. The detached leaf assessment allowed individual plants to be challenged with multiple pathotypes and their responses compared with 34 known differentials. Broad-spectrum resistance was identified within accession PL 51855, which behaved as a single major locus on crossing to three cultivars. The locus provided resistance to over 50 rust pathotypes, a greater range than seen for any of the known host resistance (Pc) genes. Strong resistance was identified in other accessions, and heterogeneity in response within accessions was common. Several accessions show multiple partial resistance responses that may be of value for developing durable resistance in cultivars. Because the sources of resistance in all but two differential lines were collected outside of Morocco, resistance in all accessions tested here are potentially novel. This study demonstrates that diversity within A. sterilis accessions collected in Morocco could be a very valuable source of resistance to crown rust, and it provides new germplasm for use in resistance breeding programs. Detached leaf assessment provides a valuable first step in the identification of promising candidates in complex gene bank accessions.