Cereal Disease Laboratory

Saint Paul, MN, United States

Cereal Disease Laboratory

Saint Paul, MN, United States

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Hiebert C.W.,Agriculture and Agri Food Canada | Kolmer J.A.,University of Minnesota | Kolmer J.A.,Cereal Disease Laboratory | McCartney C.A.,Agriculture and Agri Food Canada | And 7 more authors.
PLoS ONE | Year: 2016

Stem rust, caused by Puccinia graminis (Pgt), is a damaging disease of wheat that can be controlled by utilizing effective stem rust resistance genes. 'Thatcher' wheat carries complex resistance to stem rust that is enhanced in the presence of the resistance gene Lr34.The purpose of this study was to examine APR in 'Thatcher' and look for genetic interactions with Lr34. A RIL population was tested for stem rust resistance in field nurseries in Canada, USA, and Kenya. BSA was used to find SNP markers associated with reduced stem rust severity. A major QTL was identified on chromosome 3BL near the centromere in all environments. Seedling testing showed that Sr12 mapped to the same region as the QTL for APR. The SNP markers were physically mapped and the region carrying the resistance was searched for sequences with homology to members of the NB-LRR resistance gene family. SNP marker from one NB-LRR-like sequence, NB-LRR3 co-segregated with Sr12.Two additional populations, including one that lacked Lr34, were tested in field nurseries. NB-LRR3 mapped near the maximum LOD for reduction in stem rust severity in both populations. Lines from a population that segregated for Sr12 and Lr34 were tested for seedling Pgt biomass and infection type, as well as APR to field stem rust which showed an interaction between the genes. We concluded that Sr12, or a gene closely linked to Sr12, was responsible for 'Thatcher'-derived APR in several environments and this resistance was enhanced in the presence of Lr34. © This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.


Gill U.,Washington State University | Gill U.,Samuel Roberts Noble Foundation | Brueggeman R.,North Dakota State University | Nirmala J.,Washington State University | And 4 more authors.
Theoretical and Applied Genetics | Year: 2016

Key message: This study describes the generation, screening, genetic and molecular characterization, and high-resolution mapping of barley mutants susceptible to stem rust (Puccinia graminisf. sp.tritici) races MCCF and HKHJ.Abstract: A single gene, Rpg1, has protected barley cultivars against many races of stem rust pathogen (Puccinia graminis f. sp. tritici) for the last 70 years in the United States and Canada. To identify signaling components of protein product RPG1, we employed a mutagenesis approach. Using this approach, six mutants exhibiting susceptibility to Puccinia graminis f. sp. tritici races MCCF and HKHJ were identified in the gamma irradiated M2 population of resistant cultivar Morex, which carries Rpg1 on chromosome 7H. The mutants retained a functional Rpg1 gene and an apparently functional protein, suggesting that the mutated genes were required for downstream or upstream signaling. Selected mutants were non-allelic, hence each mutant represents a unique gene. Low and high-resolution genetic mapping of the rpr2 mutant identified chromosome 6H (bin 6) as the location of the mutated gene. The target region was reduced to 0.6 cM and gene content analyzed. Based on the published barley genomic sequence, the target region contains approximately 157 genes, including a set that encodes putative leucine-rich receptor-like protein kinases, which may be strong candidates for the gene of interest. Overall, this study presents a strong platform for future map-based cloning of genes identified in this mutant screen. © 2016 Springer-Verlag Berlin Heidelberg


Babiker E.M.,Small Grains and Potato Germplasm Research Unit | Gordon T.C.,Small Grains and Potato Germplasm Research Unit | Chao S.,Cereal Crops Research Unit | Rouse M.N.,Cereal Disease Laboratory | And 5 more authors.
Theoretical and Applied Genetics | Year: 2016

Key message: A gene for Ug99 resistance from wheat landrace CItr 4311 was detected on the long arm of chromosome 2B.Abstract: Wheat landrace CItr 4311 has seedling resistance to stem rust caused by Puccinia graminis f. sp. tritici race TTKSK and field resistance to the Ug99 race group. Parents, F1 seedlings, 121 doubled haploid (DH) lines, and 124 recombinant inbred lines (RILs) developed from a cross between CItr 4311 and the susceptible line LMPG-6 were evaluated for seedling resistance to race TTKSK. Goodness-of-fit tests indicated that a single dominant gene in CItr 4311 conditioned the TTKSK resistance. The 90 K wheat iSelect SNP platform was used to genotype parents and the DH population. The seedling resistance locus was mapped to the chromosome arm 2BL. Parents and the DH population were evaluated for field resistance in Kenya. One major QTL for the field resistance was consistently detected in the same region on 2BL as the seedling resistance. Using KASP assays, five linked SNP markers were used to verify the result in the 124 RIL, 35 wheat accessions, 46 DH lines from the LMPG-6/PI 165194 cross and F1 seedlings, and susceptible bulks derived from crosses between six resistant landraces with LMPG-6. Race specificity, mapping results, and haplotype similarity with lines with Sr9h (Gabo 56, Timstein, and PI 670015), support the hypothesis that the Sr gene in CItr 4311 and the landraces is Sr9h. The KASP assays developed in this study will be useful for pyramiding the TTKSK resistance from CItr 4311 with other Sr genes effective against Ug99. © 2016 Springer-Verlag Berlin Heidelberg (outside the USA)


Basnet B.R.,International Maize and Wheat Improvement Center | Singh S.,International Maize and Wheat Improvement Center | Lopez-Vera E.E.,CINVESTAV | Huerta-Espino J.,INIFAP CEVAMEX | And 4 more authors.
Phytopathology | Year: 2015

This study reports the identification of a new gene conferring resistance to the Ug99 lineage of races of Puccinia graminis f. sp. tritici in wheat (Triticum aestivum L.). Because the virulent races of stem rust pathogen continue to pose a serious threat in global wheat production, identification and molecular characterization of new resistance genes remains of utmost important to enhance resistance diversity and durability in wheat germplasm. Advanced wheat breeding line 'ND643/2∗ Weebill1' carries a stem rust resistance gene, temporarily designated as SrND643, effective against the Ug99 group of P. graminis f. sp. tritici races at both seedling and adult growth stages. This study was conducted to map the chromosomal location of SrND643 and identify closely linked molecular markers to allow its selection in breeding populations. In total, 123 recombinant inbred lines, developed by crossing ND643/2∗ Weebill1 with susceptible line 'Cacuke', were evaluated for stem rust response in field nurseries at Njoro, Kenya, during two growing seasons in 2010, and were genotyped with DNA markers, including Diversity Arrays Technology, simple sequence repeats (SSR), and single-nucleotide polymorphisms. Linkage mapping tagged SrND643 at the distal end of chromosome 4AL, showing close association with SSR markers Xgwm350 (0.5 centimorgans [cM]), Xwmc219 (4.1 cM), and Xwmc776 (2.9 cM). The race specificity of SrND643 is different from that of Sr7a and Sr7b, indicating that the resistance is conferred by a gene at a new locus or by a new allele of Sr7. The flanking markers Xgwm350 and Xwmc219 were predictive of the presence of SrND643 in advanced germplasm, thus validating the map location and their use in marker-assisted selection. © 2015 The American Phytopathological Society.


Olivera P.D.,University of Minnesota | Jin Y.,Cereal Disease Laboratory | Rouse M.,Cereal Disease Laboratory | Badebo A.,Ethiopian Institute of Agricultural Research | And 3 more authors.
Plant Disease | Year: 2012

North American durum lines, selected for resistance to TTKSK (Ug99) and related races of Puccinia graminis f. sp. tritici in Kenya, became susceptible in Debre Zeit, Ethiopia, suggesting the presence of stem rust races that were virulent to the TTKSK-effective genes in durum. The objective of this study was to characterize races of P. graminis f. sp. tritici present in the Debre Zeit, Ethiopia stem rust nursery. Three races of P. graminis f. sp. tritici were identified from 34 isolates: JRCQC, TRTTF, and TTKSK. Both races JRCQC and TRTTF possess virulence on stem rust resistance genes Sr13 and Sr9e, which may explain why many TTKSK-resistant durum lines tested in Kenya became susceptible in Debre Zeit. The Sr9e-Sr13 virulence combination is of particular concern because these two genes constitute major components of stem rust resistance in North American durum cultivars. In addition to Sr9e and Sr13 virulence, race TRTTF is virulent to at least three stem rust resistance genes that are effective to race TTKSK, including Sr36, SrTmp, and resistance conferred by the 1AL.1RS rye translocation. Race TRTTF is the first known race with virulence to the stem rust resistance carried by the 1AL.1RS translocation, which represents one of the few effective genes against TTKSK in winter wheat cultivars in the United States. Durum entries exhibiting resistant to moderately susceptible infection response at the Debre Zeit nursery in 2009 were evaluated for reaction to races JRCQC, TRTTF, and TTKSK at the seedling stage. In all, 47 entries were resistant to the three races evaluated at the seedling stage, whereas 26 entries exhibited a susceptible reaction. These results suggest the presence of both major and adult plant resistance genes, which would be useful in durumwheat breeding programs. A thorough survey of virulence in the population of P. graminis f. sp. tritici in Ethiopia will allow characterization of the geographic distribution of the races identified in the Debre Zeit field nursery. © 2012 The American Phytopathological Society.


Newcomb M.,U.S. Department of Agriculture | Acevedo M.,North Dakota State University | Bockelman H.E.,U.S. Department of Agriculture | Brown-Guedira G.,U.S. Department of Agriculture | And 8 more authors.
Plant Disease | Year: 2013

Wheat landraces provide a source of genetic variability for breeding. The emergence and spread of highly virulent races of the stem rust pathogen (Ug99 race group of Puccinia graminis f. sp. tritici) threaten wheat production globally. Spring wheat landraces were screened for resistance in eight field seasons at the Kenya Agricultural Research Institute, Njoro, where the Ug99 race group has become endemic. Accessions showing resistance in one season were retested and screened with molecular markers associated with resistance genes Sr2, Sr24, Sr36, and Lr34/Yr18; two height-reducing genes; and a photoperiod insensitivity allele. Of 2,509 accessions tested, 278 were categorized as resistant based on results from at least two seasons. Of these resistant accessions, 32 were positive for one or more markers for Sr2, Sr36, Rht-B1b, or Rht-D1b, indicating that they do not fit the definition of "landrace" because these genes were likely introduced via modern breeding practices. Thus, 246 resistant "landrace" accessions were identified. Of countries with more than five tested accessions, Afghanistan, Iran, Portugal, Ethiopia, Uzbekistan, Greece, Tajikistan, Bosnia and Herzegovina, and Serbia had at least 10% of tested accessions that were resistant to the Ug99 race group. Future research will characterize the resistance to determine its novelty and incorporate novel genes into improved lines.


Tsilo T.J.,Agricultural Research Council Small Grain Institute | Kolmer J.A.,Cereal Disease Laboratory | Anderson J.A.,University of Minnesota
Phytopathology | Year: 2014

Leaf rust, caused by Puccinia triticina, is the most common and widespread disease of wheat (Triticum aestivum) worldwide. Deployment of host-plant resistance is one of the strategies to reduce losses due to leaf rust disease. The objective of this study was to map genes for adult-plant resistance to leaf rust in a recombinant inbred line (RIL) population originating from MN98550-5/MN99394-1. The mapping population of 139 RILs and five checks were evaluated in 2005, 2009, and 2010 in five environments. Natural infection occurred in the 2005 trials and trials in 2009 and 2010 were inoculated with leaf rust. Four quantitative trait loci (QTL) on chromosomes 2BS, 2DS, 7AL, and 7DS were detected. The QTL on 2BS explained up to 33.6% of the phenotypic variation in leaf rust response, whereas the QTL on 2DS, 7AL, and 7DS explained up to 15.7, 8.1, and 34.2%, respectively. Seedling infection type tests conducted with P. triticina races BBBD and SBDG confirmed that the QTL on 2BS and 2DS were Lr16 and Lr2a, respectively, and these genes were expressed in the seedling and field plot tests. The Lr2a gene mapped at the same location as Sr6. The QTL on 7DS was Lr34. The QTL on 7AL is a new QTL for leaf rust resistance. The joint effects of all four QTL explained 74% of the total phenotypic variation in leaf rust severity. Analysis of different combinations of QTL showed that the RILs containing all four or three of the QTL had the lowest average leaf rust severity in all five environments. Deployment of these QTL in combination or with other effective genes will lead to successful control of leaf rust. © 2014 The American Phytopathological Society.


Fellers J.P.,U.S. Department of Agriculture | Soltani B.M.,Agriculture and Agri Food Canada | Bruce M.,U.S. Department of Agriculture | Linning R.,Agriculture and Agri Food Canada | And 3 more authors.
BMC Genomics | Year: 2013

Background: Wheat leaf rust (Puccinia triticina Eriks; Pt) and stem rust fungi (P. graminis f.sp. tritici; Pgt) are significant economic pathogens having similar host ranges and life cycles, but different alternate hosts. The Pt genome, currently estimated at 135 Mb, is significantly larger than Pgt, at 88 Mb, but the reason for the expansion is unknown. Three genomic loci of Pt conserved proteins were characterized to gain insight into gene content, genome complexity and expansion.Results: A bacterial artificial chromosome (BAC) library was made from P. triticina race 1, BBBD and probed with Pt homologs of genes encoding two predicted Pgt secreted effectors and a DNA marker mapping to a region of avirulence. Three BACs, 103 Kb, 112 Kb, and 166 Kb, were sequenced, assembled, and open reading frames were identified. Orthologous genes were identified in Pgt and local conservation of gene order (microsynteny) was observed. Pairwise protein identities ranged from 26 to 99%. One Pt BAC, containing a RAD18 ortholog, shares syntenic regions with two Pgt scaffolds, which could represent both haplotypes of Pgt. Gene sequence is diverged between the species as well as within the two haplotypes. In all three BAC clones, gene order is locally conserved, however, gene shuffling has occurred relative to Pgt. These regions are further diverged by differing insertion loci of LTR-retrotransposon, Gypsy, Copia, Mutator, and Harbinger mobile elements. Uncharacterized Pt open reading frames were also found; these proteins are high in lysine and similar to multiple proteins in Pgt.Conclusions: The three Pt loci are conserved in gene order, with a range of gene sequence divergence. Conservation of predicted haustoria expressed secreted protein genes between Pt and Pgt is extended to the more distant poplar rust, Melampsora larici-populina. The loci also reveal that genome expansion in Pt is in part due to higher occurrence of repeat-elements in this species. © 2013 Fellers et al.; licensee BioMed Central Ltd.


Lysoe E.,Norwegian Institute for Agricultural And Environmental Research Bioforsk | Seong K.-Y.,University of Minnesota | Kistler H.C.,University of Minnesota | Kistler H.C.,Cereal Disease Laboratory
Molecular Plant-Microbe Interactions | Year: 2011

Fusarium graminearum causes head blight disease in wheat and barley. To help understand the infection process on wheat, we studied global gene expression of F. graminearum in a time series from 24 to 196 h after inoculation, compared with a noninoculated control. The infection was rapid and, after 48 h, over 4,000 fungal genes were expressed. The number of genes expressed increased over time up to 96 h (>8,000 genes), and then declined at the 144- and 192-h post-inoculation time points. After subtraction of genes found expressed on complete medium, during carbon or nitrogen starvation, and on barley, only 355 were found exclusively expressed in wheat, mostly genes with unknown function (72.6%). These genes were mainly found in single-nucleotide polymorphism-enriched islands on the chromosomes, suggesting a higher evolutionary selection pressure. The annotated genes were enriched in functional groups predicted to be involved in allantoin and allantoate transport, detoxification, nitrogen, sulfur and selenium metabolism, secondary metabolism, carbohydrate metabolism, and degradation of polysaccharides and ester compounds. Several putative secreted virulence factors were also found expressed in wheat. © 2011 The American Phytopathological Society.


Kolmer J.A.,Cereal Disease Laboratory
Plant Disease | Year: 2010

Leaf rust, caused by the fungus Puccinia triticina, is an important disease of soft red winter wheat cultivars that are grown in the southern and eastern United States. The objectives of this study were to identify the leaf rust resistance genes in two soft red winter wheat cultivars, Coker 9663 and Pioneer 26R61, that have been widely grown and were initially highly resistant to leaf rust. Both cultivars were crossed with the leaf-rust-susceptible spring wheat cv. Thatcher and the F1 plants were crossed to Thatcher to obtain backcross (BC1) F2 families. In seedlings, the Thatcher/Coker 9663 BC1F2 families segregated for three independent seedling resistance genes when tested with different leaf rust isolates. The leaf rust infection types of selected BC1F3 lines, when tested with different leaf rust isolates, indicated that seedling resistance genes Lr9, Lr10, and Lr14a were present. In field plot tests, BC1F4 lines that were seedling susceptible had some adult plant resistance to leaf rust. Seedlings of the Thatcher/Pioneer 26R61 BC1F2 families segregated for two independent resistance genes. Infection types of selected BC1F3 lines indicated the presence of Lr14b and Lr26. The adult plant gene Lr13 was determined to be present in selected BC 1F4 lines that were tested with different leaf rust isolates in greenhouse tests.

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