Browsing by Subject "Puccinia graminis f. sp. tritici"
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Item Amplified fragment length polymorphism and virulence polymorphism in Puccinia hordei(Canadian Journal of Plant Pathology, 2007) Sun, Y; Neate, S.M.; Zhong, S; Steffenson, Brian; Friesen, T. L.Puccinia hordei is the causal agent of barley leaf rust. To study the genetic diversity in P. hordei, 45 isolates with diverse virulence patterns and geographical origins were analyzed using amplified fragment length polymorphism markers. Two pathotypes of Puccinia graminis f. sp. tritici and one isolate of P. graminis f. sp. secalis were included in the analysis for comparison. Six primer-pair combinations of amplified fragment length polymorphism were used and a total of 782 polymorphic markers were generated. Cluster analysis showed that P. graminis f. sp. tritici and P. graminis f. sp. secalis were distinctly different from P. hordei. The P. hordei isolates were clustered into five groups: group I contained a single, rare isolate that was virulent on all resistance genes except Rph13 and Rph15 ; group II contained a single isolate found to be virulent on the resistance gene Rph15 ; group III contained 2 isolates; group IV contained 24 isolates, 11 from the United States and 13 from diverse locations around the world; and group V contained 17 isolates, 7 from California, 7 from other states of United States, and 3 from central Europe. The study revealed that molecular diversity in P. hordei can be associated with virulence, but not well with geographic origin.Item The barley stem rust-resistance gene Rpg1 is a novel disease-resistance gene with homology to receptor kinases(Proceedings of the National Academy of Sciences of the United States of America, 2002-07-09) Steffenson, Brian; Brueggeman, R.; Rostoks, N.; Kudrna, D.; Kilian, A.; Han, F.; Chen, J.; Druka, A.; Kleinhofs, A.Stem rust caused by Puccinia graminis f. sp. tritici was among the most devastating diseases of barley in the northern Great Plains of the U.S. and Canada before the deployment of the stem rust-resistance gene Rpg1 in 1942. Since then, Rpg1 has provided durable protection against stem rust losses in widely grown barley cultivars (cvs.). Extensive efforts to clone Rpg1 by synteny with rice provided excellent flanking markers but failed to yield the gene because it does not seem to exist in rice. Here we report the map-based cloning and characterization of Rpg1. A high-resolution genetic map constructed with 8,518 gametes and a 330-kb bacterial artificial chromosome contig physical map positioned the gene between two crossovers ≈0.21 centimorgan and 110 kb apart. The region including Rpg1 was searched for potential candidate genes by sequencing low-copy probes. Two receptor kinase-like genes were identified. The candidate gene alleles were sequenced from resistant and susceptible cvs. Only one of the candidate genes showed a pattern of apparently functional gene structure in the resistant cvs. and defective gene structure in the susceptible cvs. identifying it as the Rpg1 gene. Rpg1 encodes a receptor kinase-like protein with two tandem protein kinase domains, a novel structure for a plant disease-resistance gene. Thus, it may represent a new class of plant resistance genes.Item Changing the Game: Using Integrative Genomics to Probe Virulence Mechanisms of the Stem Rust Pathogen Puccinia graminis f. sp. tritici(Frontiers in Plant Science, 2016-02-24) Steffenson, Brian; Figueroa, Melania; Upadhyaya, Narayana M; Sperschneider, Jana; Park, Robert F; Szabo, Les J; Ellis, Jeff G; Dodds, Peter NThe recent resurgence of wheat stem rust caused by new virulent races of Puccinia graminis f. sp. tritici (Pgt) poses a threat to food security. These concerns have catalyzed an extensive global effort toward controlling this disease. Substantial research and breeding programs target the identification and introduction of new stem rust resistance (Sr) genes in cultivars for genetic protection against the disease. Such resistance genes typically encode immune receptor proteins that recognize specific components of the pathogen, known as avirulence (Avr) proteins. A significant drawback to deploying cultivars with single Sr genes is that they are often overcome by evolution of the pathogen to escape recognition through alterations in Avr genes. Thus, a key element in achieving durable rust control is the deployment of multiple effective Sr genes in combination, either through conventional breeding or transgenic approaches, to minimize the risk of resistance breakdown. In this situation, evolution of pathogen virulence would require changes in multiple Avr genes in order to bypass recognition. However, choosing the optimal Sr gene combinations to deploy is a challenge that requires detailed knowledge of the pathogen Avr genes with which they interact and the virulence phenotypes of Pgt existing in nature. Identifying specific Avr genes from Pgt will provide screening tools to enhance pathogen virulence monitoring, assess heterozygosity and propensity for mutation in pathogen populations, and confirm individual Sr gene functions in crop varieties carrying multiple effective resistance genes. Toward this goal, much progress has been made in assembling a high quality reference genome sequence for Pgt, as well as a Pan-genome encompassing variation between multiple field isolates with diverse virulence spectra. In turn this has allowed prediction of Pgt effector gene candidates based on known features of Avr genes in other plant pathogens, including the related flax rust fungus. Upregulation of gene expression in haustoria and evidence for diversifying selection are two useful parameters to identify candidate Avr genes. Recently, we have also applied machine learning approaches to agnostically predict candidate effectors. Here, we review progress in stem rust pathogenomics and approaches currently underway to identify Avr genes recognized by wheat Sr genes.Item Effect of incubation time and temperature on the phenotypic expression of rpg4 to Puccinia graminis f. sp. tritici in barley(Canadian Journal of Plant Pathology, 1997-03-01) Sun, Yonglian; Steffenson, BrianTo study the effect of incubation time and temperature on the phenotypic expression of rpg4, five barley genotypes with this resistance gene were infected with pathotype QCCJ of Puccinia graminis f. sp. tritici at the seedling stage, then subjected to various times of incubation at either 18-19°C or 27~28°C. Genotypes with rpg4 exhibited low (0, 0;, and 1), mesothetic (e.g. 3-210;, 120;3), and high (3,3) infection types at 18-19°C after initial incubation at 27-28°C for 0-28, 40-76, and 88 or more hours, respectively. A period of 88 or more hours of initial incubation at high temperature rendered the rpg4 resistance completely ineffective against this pathotype of P. g. f. sp. tritici. In contrast, high, mesothetic, and low infection types were found for the same genotypes at 27-28°C after initial incubation at 18-19°C for 0-40, 52-100, and 112 or more hours, respectively. The resistant infection types conferred by rpg4 are apparently established within the first 112 hours after the end of the infection period since subsequent shifts to higher temperature did not result in marked changes in the resistance response. These data indicate the critical importance of maintaining precise temperature control when assessing the infection phenotypes of barley genotypes carrying the stem rust resistance gene rpg4.Item Genetically engineered stem rust resistance in barley using the Rpg1 gene(Proceedings of the National Academy of Sciences of the United States of America, 2003-01-07) Steffenson, Brian; Horvath, Henriette; Rostoks, Nils; Brueggeman, Robert; Wettstein, Diter von; Kleinhofs, AndrisThe stem-rust-susceptible barley cv. Golden Promise was transformed by Agrobacterium-mediated transformation of immature zygotic embryos with the Rpg1 genomic clone of cv. Morex containing a 520-bp 5′ promoter region, 4,919-bp gene region, and 547-bp 3′ nontranscribed sequence. Representatives of 42 transgenic barley lines obtained were characterized for their seedling infection response to pathotype Pgt-MCC of the stem rust fungus Puccinia graminis f. sp. tritici. Golden Promise was converted from a highly susceptible cultivar into a highly resistant one by transformation with the dominant Rpg1 gene. A single copy of the gene was sufficient to confer resistance against stem rust, and progenies from several transformants segregated in a 3:1 ratio for resistance/susceptibility as expected for Mendelian inheritance. These results unequivocally demonstrate that the DNA segment isolated by map-based cloning is the functional Rpg1 gene for stem rust, resistance. One of the remarkable aspects about the transformants is that they exhibit a higher level of resistance than the original sources of Rpg1 (cvs. Chevron and Peatland). In most cases, the Golden Promise transformants exhibited a highly resistant reaction where no visible sign of infection was evident. Hypersensitive necrotic “fleck” reactions were also observed, but less frequently. With both infection types, pathogen sporulation was prevented. Southern blot and RT-PCR analysis revealed that neither Rpg1 gene copy number nor expression levels could account for the increased resistance observed in Golden Promise transformants. Nevertheless, this research demonstrates that stem-rust-susceptible barley can be made resistant by transformation with the cloned Rpg1 gene.Item New resistance in old places: resistance to the Ug99 race group of Puccinia graminis f. sp. tritici in wheat intra/inter-generic hybrids and historic germplasm(2015-01) Kielsmeier-Cook, JoshuaWheat stem rust caused by the rust fungus, Puccinia graminis f. sp. tritici, threatens global wheat (Triticum aestivum) production. New races originating in Eastern Africa have overcome many existing stem rust resistance genes. The W. J. Sando collection of wheat intra/inter-generic hybrids is a valuable source of stem rust resistance. The entire collection was characterized for seedling stem rust resistance to 8 races of the stem rust pathogen and cytogenetic analysis was performed on select lines. Several accessions are postulated to contain new sources of resistance. Full screening results are displayed in Supplementary Table S1 and the pedigrees of 29 resistant lines are displayed in Supplementary Table S2. South African accession PI 410954 displayed strong resistance to stem rust race TTKSK at the seedling stage and under field conditions. The source of new resistance was located and material suitable for integration into modern spring wheat breeding programs was produced.Item Proteolysis of the barley receptor-like protein kinase RPG1 by a proteasome pathway is correlated with Rpg1-mediated stem rust resistance(Proceedings of the National Academy of Sciences of the United States of America, 2007-06-12) Steffenson, Brian; Nirmala, Jayaveeramuthu; Dahl, Stephanie; Kannangara, C.Gamini; Wettstein, Diter von; Chen, Xianming; Kleinhofs, AndrisIn plants, disease resistance mediated by the gene-for-gene mechanism involves the recognition of specific effector molecules produced by the pathogen either directly or indirectly by the resistance-gene products. This recognition triggers a series of signals, thereby serving as a molecular switch in regulating defense mechanisms by the plants. To understand the mechanism of action of the barley stem rust resistance gene Rpg1, we investigated the fate of the RPG1 protein in response to infection with the stem rust fungus, Puccinia graminis f. sp. tritici. The investigations revealed that RPG1 disappears to undetectable limits only in the infected tissues in response to avirulent, but not virulent pathotypes. The RPG1 protein disappearance is rapid and appears to be due to specific protein degradation via the proteasome-mediated pathway as indicated by inhibition with the proteasomal inhibitor MG132, but not by other protease inhibitors.Item Quantitative and Qualitative Stem Rust Resistance Factors in Barley Are Associated with Transcriptional Suppression of Defense Regulons(PLoS Genetics, 2011-07-28) Steffenson, Brian; Moscou, Matthew J; Lauter, Nick; Wise, Roger PStem rust (Puccinia graminis f. sp. tritici; Pgt) is a devastating fungal disease of wheat and barley. Pgt race TTKSK (isolate Ug99) is a serious threat to these Triticeae grain crops because resistance is rare. In barley, the complex Rpg-TTKSK locus on chromosome 5H is presently the only known source of qualitative resistance to this aggressive Pgt race. Segregation for resistance observed on seedlings of the Q21861 × SM89010 (QSM) doubled-haploid (DH) population was found to be predominantly qualitative, with little of the remaining variance explained by loci other than Rpg-TTKSK. In contrast, analysis of adult QSM DH plants infected by field inoculum of Pgt race TTKSK in Njoro, Kenya, revealed several additional quantitative trait loci that contribute to resistance. To molecularly characterize these loci, Barley1 GeneChips were used to measure the expression of 22,792 genes in the QSM population after inoculation with Pgt race TTKSK or mock-inoculation. Comparison of expression Quantitative Trait Loci (eQTL) between treatments revealed an inoculation-dependent expression polymorphism implicating Actin depolymerizing factor3 (within the Rpg-TTKSK locus) as a candidate susceptibility gene. In parallel, we identified a chromosome 2H trans-eQTL hotspot that co-segregates with an enhancer of Rpg-TTKSK-mediated, adult plant resistance discovered through the Njoro field trials. Our genome-wide eQTL studies demonstrate that transcript accumulation of 25% of barley genes is altered following challenge by Pgt race TTKSK, but that few of these genes are regulated by the qualitative Rpg-TTKSK on chromosome 5H. It is instead the chromosome 2H trans-eQTL hotspot that orchestrates the largest inoculation-specific responses, where enhanced resistance is associated with transcriptional suppression of hundreds of genes scattered throughout the genome. Hence, the present study associates the early suppression of genes expressed in this host–pathogen interaction with enhancement of R-gene mediated resistance.Item Reaction of barley seedlings with different stem rust resistance genes to Puccinia graminis f. sp. tritici and P. g. f. sp. secalis(Canadian Journal of Plant Pathology, 2005-03-01) Sun, Y; Steffenson, BrianSeedling tests are desirable for disease resistance screening in breeding programs because a large number of lines can be evaluated in a short period of time and with modest space requirements. This study was undertaken to identify pathotypes of the wheat stem rust pathogen (Puccinia graminis f. sp. tritici) and cultures of the rye stem rust pathogen (Puccinia graminis f. sp. secalis) that might be useful for detecting resistance genes in barley (Hordeum vulgare) at the seedling stage. Barley accessions with different resistance genes (Rpg1, Rpg2, Rpg3, rpg4, and rpgBH) were assessed for their infection types (ITs) to 13 pathotypes of P. graminis f. sp. tritici and two cultures of P. graminis f. sp. secalis at two temperature regimes (18–21 °C and 24–25 °C). The P. graminis f. sp. tritici pathotypes HKHJ and QCCJ were effective for specifically detecting Rpg1 and rpg4, respectively, and will facilitate resistance breeding efforts. No cultures were found to be effective for specifically detecting the resistance of Hietpas-5, PI 382313, or Black Hulless at the seedling stage. Some pathotypes or cultures did, however, produce low ITs on these sources and will therefore be useful for genetic studies involving crosses with susceptible barleysItem Receptivity of barley to Puccinia graminis f. sp. tritici(Canadian Journal of Plant Pathology, 2009-12-29) Steffenson, Brian; Wilcoxson, RoyThe receptivity of barley genotypes {Hordeum vulgare) was studied in seedlings and adult plants in the greenhouse and in adult plants in the field to races I13-RTQ and 151-QSH of Puccinia graminis f. sp. tritici. In the greenhouse, significant differences in number of uredia/cm2 of leaf were detected due to the effects of races, host genotypes, and their interaction. The cultivar Hiproly was most receptive (had the most uredia) and 80-TT-29 was least receptive (had the fewest uredia) to both races at both growth stages. With race 151-QSH, genotypes with the T-gene, 80-TT-29 and Manker had low weighted infection types (seedling stage), moderately resistant host responses (adult stage), and lower receptivity (both growth stages) than cultivars lacking this gene. With race 113-RTQ, the T-gene was associated with low receptivity only in 80-TT-29. The data suggest that gene(s) other than the T-gene may confer receptivity to P. graminis f. sp. tritici. The ranking of genotypes and the relative differences in receptivity were similar in seedling and adult plants. In the field, genotypes with the T-gene had mostly moderately resistant reactions and fewer uredia than those without the gene. The significant race x host genotype interaction in this study suggests that receptivity in barley varies due to the specific host-parasite combination.Item The rpg4/Rpg5 stem rust resistance locus in barley; resistance genes and cytoskeleton dynamics(Cell Cycle, 2009-04-01) Brueggeman, Robert; Steffenson, Brian; Kleinhofs, AndrisTwo closely linked resistance genes, rpg4 and Rpg5, conferring resistance to several races of Puccinia graminis, were cloned and characterized. The Rpg5 gene confers resistance to an isolate of Puccinia graminis f. sp. secalis (Pgs), while rpg4 confers resistance to Puccinia graminis f. sp. tritici (Pgt). Rpg5 is a novel gene containing nucleotide binding site-leucine rich repeat domains in combination with a serine threonine protein kinase domain. High-resolution mapping plus allele and recombinant sequencing identified the rpg4 gene, which encodes an actin depolymerizing factor-like protein (ADF2). Resistance against the Pgt races QCCJ, MCCF, TTKSK (aka Ug99) and RCRS requires both Rpg5 and rpg4, while Rpg5 alone confers resistance to Pgs isolate 92-MN-90. The dependency on the actin modifying protein ADF2 indicates cytoskeleton reorganization or redirection plays a role in pathogen-host interactions. Rpg5 may interact with ADF2 to activate or deactivate its function in the resistance response. Alternatively, Rpg5 could initiate signal transduction leading to resistance in response to detecting ADF2 protein modification. Pgt may redirect the actin cytoskeleton by inducing modifications of ADF2. The redirection of actin could possibly enable the pathogen to develop a haustoria-plant cell cytoskeleton interface for acquisition of nutrients.Item SNP data of 18SPA092-1 and 25 reference isolates of the wheat stem rust fungus, Puccinia graminis f. sp. tritici(2022-02-04) Olivera Firpo, Pablo D; Villegas, Dolors; Cantero-Martinez, Carlos; Szabo, Les J; Rouse, Matthew N; Luster, Douglas; Bartaula, Radhika; Lopes, Marta; Jin, Yue; oliv0132@umn.edu; Olivera Firpo, Pablo D; Wheat stem rust research project / USDA-ARS Cereal Disease Laboratory and University of MinnesotaThis dataset includes 1838 SNP loci of 26 isolates of the wheat stem rust fungus, Puccina graminis f .sp. tritici. This data was used to assess the genetic relationship of a unique fungal isolate (18SPA092-1, race TKHBK) derived from a wheat stem rust sample collected in Spain in 2018 with a set of 25 reference isolates from previously defined genetic clades. Results indicate that isolate 18SPA092-1 had a unique multilocus genotype and constitutes a previously unknown lineage. Data is being released now in conjunction with publication of a primary research paper that describes this work.Item SNP data of 56 Kazakhstani and 23 reference isolates of the wheat stem rust fungus, Puccinia graminis f. sp. tritici(2022-03-09) Olivera Firpo, Pablo; Szabo, Les J; Kokhmetova, Alma; Morgunov, Alexey; Luster, Douglas; Jin, Yue; oliv0132@umn.edu; Olivera Firpo, Pablo, D; Wheat stem rust research project | USDA ARS Cereal Disease Lab and University of MinnesotaThis dataset includes 2310 SNP loci of 79 isolates of the wheat stem rust fungus, Puccina graminis f .sp. tritici (Pgt). This data was used to assess the genetic diversity of a Pgt population (56 isolates) from Kazakhstan derived from stem rust samples collected in 2015-2017 wheat growing seasons. Twenty-three reference isolates from previously defined genetic clades were included in the analysis. Results indicate that the Pgt population form Kazakhstan is highly diverse and most of the isolates are of sexual origin. Data is being released now in conjunction with publication of a primary research paper that describes this work.Item Sources of resistance to pathotype QCC of Puccinia graminis f. sp. tritici in barley(Crop Science, 1994) Steffenson, Brian; Jin, Yue; Fetch, Thomas G.The occurrence of a wheat stem rust (Puccinia graminis Pers.:Pers. f. sp. tritici Eriks. & E. Henn.) pathotype (Pgt-QCC) with virulence for the Rpg1 gene in barley (Hordeum vulgare L.) necessitated the search for resistant barley germplasm. From preliminary screenings of over 18 000 barley accessions, 13 lines were identified as possessing resistance to pathotype QCC: ‘Diamond’, ‘Hietpas 5’, Q21861, PC 11, PC 84, PC 249, PC 250, CI 5541, PI 452406, PI 452421, PI 477843, PI 477854, and PI 477860. This study was conducted to further characterize the reaction of the selected lines to pathotype QCC. The reaction was assessed by evaluating infection types at the seedling stage and infection responses at the adult plant stage in the greenhouse, and by evaluating disease severity and infection responses at the adult plant stage in the field compared to susceptible cultivars. Most lines exhibited low to intermediate infection types at the seedlings stage and moderately resistant to moderately susceptible infection responses at the adult plant stage in the greenhouse experiments. Among the selected lines, Q21861 exhibited the highest level of resistance at both the seedling and adult plant stages. These lines may provide an adequate level of resistance to pathotype QCC for cultivar development.