The immune Rysto receptor recognizes a broadly conserved feature of potyviral coat proteins

Potyviruses are the largest group of plant RNA viruses, causing significant losses in many crops. Among them, potato virus Y (PVY) is particularly important, and enhances the severity of infections by other viruses. The Rysto gene confers PVY resistance and encodes a TIR-NLR intracellular immune receptors that recognizes PVY coat protein (CP). To define a minimal CP fragment sensed by Rysto, we created a series of truncated CP variants and expressed these CP derivatives in Rysto transgenic plants. Deletions that affect the 149 amino acid CP core region lose the ability to trigger Rysto-dependent defence activation. Furthermore, point mutations in the amino acid residues Ser126, Arg157, and Asp201 of the highly conserved RNA-binding pocket of potyviral CP, reduce or abolish Rysto-dependent responses, demonstrating that appropriate folding of the CP core is required for Rysto-mediated recognition. Consistent with these data, we found Rysto recognises CPs of various viruses that share a similar core region, but not those lacking it. Finally, we demonstrated that Rysto provides immunity to plum pox virus and turnip mosaic virus, demonstrating its wide range of applications in disease-resistant crop engineering. In parallel, we showed that CP triggered Rysto activation is SAG101- but not PAD4- or SA- level dependent. Our findings shed new light on how R proteins can detect viruses by sensing highly conserved structural patterns.

Genome Enhanced Marker Improvement for Potato Virus Y Disease Resistance in Potato

Potato is an important food crop worldwide and is grown in a large number of countries. As such, the crop is under disease pressures and the need for selecting disease resistance genes during breeding programs is essential. Of particular importance within Australia and other parts of the world is the potyvirus, Potato virus Y (PVY). In this paper, three commonly used PVY resistance markers, M45, RYSC3 and M6, were evaluated using existing genomic resources and phenotypic data from the Australian potato breeding program to identify a region where the PVY resistance gene, Ryadg may reside. A region of Chromosome XI was investigated, and a cluster of disease resistance genes was identified that the resistance gene Ryadg is suspected to reside within. Protein characterization was also performed on the putative resistant gene. A specific variant that had complete association with the resistance gene was identified and a single nucleotide polymorphism (SNP) assay was designed to avoid dissociation of marker and gene in future breeding programs. This SNP marker (SNP37279) was validated as a Kompetitive Allele-specific PCR (KASP) genotyping assay and was found to perform more accurately than all previously used markers for detecting Ryadg.

Screening for PVYN-Wi Resistance in Kenyan Potato Cultivars

Over the past decades, two new strains of potato virus Y (PVYN-Wi (Wilga) and PVYNTN) have been the most widely distributed worldwide in potato crops (Solanum tuberosum L.). Effective resistance genes comprehensive against all the known strains of the virus are available in some commercial cultivars. However, not all resistance genes in cultivars have been challenged by the new PVY strains to confirm effectiveness of the resistances available. This study was set to identify PVY resistant genotypes and estimate yield losses in potato cultivars in Kenya. PVYN-Wi was used for PVY resistance screening because it was the most prevalent strain in Kenya and also was found to produce more severe infection in Kenyan potato genotypes compared with PVYO (the second most prevalent strain). Three cultivars, Kenya Karibu, Sherekea and Unica, were identified as resistant because they did not get PVY infections over three seasons of both greenhouse and field screening experiments. In addition, two early generation breeding clones were identified as resistant at 28 days post-inoculation. In susceptible cultivars Dutch Robyjn, Nyayo and Kenya Mpya, PVYN-Wi infection caused yield losses of 21.6, 39.0 and 53.1%, respectively, after three seasons. This study demonstrated the necessity to screen for PVY resistance based on the most prevalent PVY strains and to make an informed decision on the usefulness of current levels of resistance in potato cultivars.

Marker assisted selection of potato breeding lines with combination of PVY resistance genes from different wild species

Potato virus Y (PVY) is considered as one of the most harmful virus infections of this crop. Thus, it is a topical problem to breed potato varieties resistant against a wide range of PVY strains and to create initial breeding material that will have a combination of resistance genes from different species. The aim of the study was: (1) to genotype a collection of 376 breeding lines (BL), developed from complex interspecific hybrids, using DNA markers of PVY resistance genes, (2) to identify accessions with markers of resistance genes from different species for subsequent use in marker assisted selection (MAS), (3) to evaluate the suitability of DNA markers of PVY resistance genes for genotyping BL developed through interspecific hybridization. It was ascertained that the markers most widely represented in the collection were RYSC3 of the Ryadg gene (49.7%), Ry364 and RAPD38-530 of the Rychc gene (50.5% and 45.2%, respectively), and Yes3-3A of the Rysto gene (29.8%). The markers Ry186 of Rychc and GP122/EcoRV780 of Ryf-sto were found only in some accessions. The frequency of occurrence of BL that had markers of PVY resistance genes from two different species varied between 2.7% (Yes3-3a marker of Rysto and both two markers of Rychc) and 8.5-9.0% (RYSC3 marker of Ryadg and both two markers of Rychc, or only Ry364 marker of this gene). In total, the collection was found to contain 134 BL (47.6%) with markers of resistance genes from two different species. A combination of four markers for three genes of different origin (Ryadg, Rysto and Rychc) was found in 27 BL (7.2%). Extreme resistance to PVY of most BL (302 out of 357) was obviously determined by the presence in them of the currently used resistance genes detected by DNA markers applied in the study. Nevertheless, a significant part of accessions (55 of 61) that did not have any markers was resistant to PVY. At the same time, 13 BL (3.5%) with the markers were susceptible to the virus. Such a level of discrepancies is considered as acceptable for the initial MAS of breeding material. The obtained data on the presence of the markers of PVY resistance genes of different origin and their combination in BL ensures a more effective use of such BL in breeding in comparison with the BL resistant to the virus, though lacking corresponding markers.