Sr36- and Sr5-Mediated Resistance Response to Puccinia graminis f. sp. tritici Is Associated with Callose Deposition in Wheat Guard Cells.

Race-specific resistance of wheat to Puccinia graminis f. sp. tritici is primarily posthaustorial and often involves the induction of a hypersensitive response (HR). The aim of this study was to investigate host defense responses induced in interactions between P. graminis f. sp. tritici races and wheat lines carrying different race-specific stem rust resistance (Sr) genes. In incompatible interactions between wheat lines carrying Sr36 in three genetic backgrounds (LMPG, Prelude, or W2691) and avirulent P. graminis f. sp. tritici races MCCFC or RCCDM, callose accumulated within 24 h in wheat guard cells contacted by a P. graminis f. sp. tritici appressorium, and P. graminis f. sp. tritici ingress was inhibited following appressorium formation. Accordingly, the expression of transcripts encoding a callose synthase increased in the incompatible interaction between LMPG-Sr36 and avirulent P. graminis f. sp. tritici race MCCFC. Furthermore, the inhibition of callose synthesis through the infiltration of 2-deoxy-D-glucose (DDG) increased the ability of P. graminis f. sp. tritici race MCCFC to infect LMPG-Sr36. A similar induction of callose deposition in wheat guard cells was also observed within 24 h after inoculation (hai) with avirulent P. graminis f. sp. tritici race HKCJC on LMPG-Sr5 plants. In contrast, this defense response was not induced in incompatible interactions involving Sr6, Sr24, or Sr30. Instead, the induction of an HR and cellular lignification were noted. The manifestation of the HR and cellular lignification was induced earlier (24 hai) and was more extensive in the resistance response mediated by Sr6 compared with those mediated by Sr24 or Sr30. These results indicate that the resistance mediated by Sr36 is similar to that mediated by Sr5 but different from those triggered by Sr6, Sr24, or Sr30. Resistance responses mediated by Sr5 and Sr36 are prehaustorial, and are a result of very rapid recognition of molecules derived from avirulent isolates of P. graminis f. sp. tritici, in contrast to the responses triggered in lines with Sr6, Sr24, and Sr30.

Analysis of Stem Rust Resistance in Australian Barley Cultivars.

Eighty-two Australian and five exotic barley cultivars were evaluated at the seedling stage for resistance to the Australian stem rust pathotype 98-1,2,3,5,6. Although most of these cultivars exhibited mesothetic (mixed infection type) reactions that were associated with a high level of chlorosis, two ('O'Connor' and 'Pacific Ranger') were highly resistant. Marker analysis indicated that four Australian cultivars ('Empress', 'Vlamingh', Pacific Ranger, and 'Yerong') possess the stem rust resistance gene Rpg1. Tests conducted using North American Puccinia graminis f. sp. tritici pathotypes MCCJ and QCCJ supported marker results and indicated that 'Pacific Ranger' and 'Vlamingh' likely carry additional stem rust resistance genes. Based on pedigree information and results from multipathotype tests, these genes are believed to be uncharacterized and, therefore, new. The resistance in Australian barley 'Franklin' conferred resistance against all pathotypes tested in this study. Studies of inheritance to MCCJ revealed that it possessed an unknown seedling resistance, which was independent of and displayed additivity to Rpg1.

Identification and mapping in spring wheat of genetic factors controlling stem rust resistance and the study of their epistatic interactions across multiple environments.

Stem rust (Puccinia graminis f. sp. tritici) is responsible for major production losses in hexaploid wheat (Triticum aestivum L.) around the world. The spread of stem rust race Ug99 and variants is a threat to worldwide wheat production and efforts are ongoing to identify and incorporate resistance. The objectives of this research were to identify quantitative trait loci (QTL) and to study their epistatic interactions for stem rust resistance in a population derived from the Canadian wheat cultivars AC Cadillac and Carberry. A doubled haploid (DH) population was developed and genotyped with DArT(®) and SSR markers. The parents and DH lines were phenotyped for stem rust severity and infection response to Ug99 and variant races in 2009, 2010 and 2011 in field rust nurseries near Njoro, Kenya, and to North American races in 2011 and 2012 near Swift Current, SK, Canada. Seedling infection type to race TTKSK was assessed in a bio-containment facility in 2009 and 2012 near Morden, MB. Eight QTL for stem rust resistance and three QTL for pseudo-black chaff on nine wheat chromosomes were identified. The phenotypic variance (PV) explained by the stem rust resistance QTL ranged from 2.4 to 48.8 %. AC Cadillac contributed stem rust resistance QTL on chromosomes 2B, 3B, 5B, 6D, 7B and 7D. Carberry contributed resistance QTL on 4B and 5A. Epistatic interactions were observed between loci on 4B and 5B, 4B and 7B, 6D and 3B, 6D and 5B, and 6D and 7B. The stem rust resistance locus on 6D interacted synergistically with 5B to improve the disease resistance through both crossover and non-crossover interactions depending on the environment. Results from this study will assist in planning breeding for stem rust resistance by maximizing QTL main effects and epistatic interactions.

Molecular cytological characterization of two novel durum--Thinopyrum intermedium partial amphiploids with resistance to leaf rust, stem rust and Fusarium head blight.

Thinopyrum intermedium, a wild relative of wheat, is an excellent source of disease resistance. Two novel partial amphiploids, 08-47-50 and 08-53-55 (2n = 6x = 42), were developed from wide crosses between durum wheat and Th. intermedium. Meiotic analysis showed that pollen mother cells of the two partial amphiploids formed an average 20.49 bivalents for 08-47-50 and 20.67 bivalents for 08-53-55, indicating that they are basically cytologically stable. GISH analysis revealed that the two partial amphiploids carried different chromosome compositions. 08-47-50 had fourteen chromosomes from Th. intermedium and its alien chromosomes included six St-, four E(e) - and four E(e)-St translocated chromosomes, whereas 08-53-55 had four St- and ten E(e)-St translocated chromosomes. Fungal disease evaluation indicated that both partial amphiploids had a high level of resistance to FHB, leaf rust and stem rust race Ug99. These two novel partial amphiploids with multiple disease resistance could be used as a new source of multiple disease resistance in bread wheat and durum wheat breeding programs.

Races of Puccinia graminis f. sp. tritici with Combined Virulence to Sr13 and Sr9e in a Field Stem Rust Screening Nursery in Ethiopia.

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 durum-wheat-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.

First Report of Puccinia graminis f. sp. tritici Races with Virulence to Wheat Stem Rust Resistance Genes Sr31 and Sr24 in Eritrea.

Wheat stem rust, caused by Puccinia graminis f. sp. tritici, has historically been a major limiting factor in wheat production. Identification of isolate Ug99 in Uganda in 1999 highlighted the vulnerability of a large proportion of the global wheat crop and raised international concerns. Since initial detection, seven races have been identified in the Ug99 lineage and occurrence has been confirmed in nine countries (4). During rust surveys of Eritrea undertaken in October 2010, stem rust was found to be widespread throughout the highland wheat-growing areas. Presence of P. graminis f. sp. tritici was recorded in 95% of the 92 cereal fields surveyed, with high disease severity (>40%) recorded at 50 sites. Collected stem rust samples were analyzed for race identity in a level 3 biocontainment laboratory in Canada. Nine collections yielded viable spores for infection studies. Virulence analysis with 20 differentials in the letter-code nomenclature system (1) identified two races from repeated experiments; TTKST (four confirmed isolates) and PTKST (five confirmed isolates). Both races belong to the Ug99 lineage and both exhibit combined Sr31 and Sr24 virulence. TTKST and PTKST differ only in their virulence or avirulence to Sr21, respectively. This first confirmation of TTKST and PTKST in Eritrea is important because it represents further geographical spread of Ug99-related races. Since first detection of a Sr24 variant of Ug99 (race TTKST) in Kenya in 2006 (1), these variants have become the predominant P. graminis f. sp. tritici pathotypes in most of eastern Africa. Race TTKST caused epidemics in Kenya in 2007, and race PTKST was first detected in Ethiopia the same year (T. Fetch, unpublished data). Recent detection of race PTKST in three additional southern Africa countries (South Africa, Zimbabwe, and Mozambique [2,3]) indicates on-going range expansion within the African continent. Sr24-virulent variants of Ug99 are a cause for concern since a high frequency of cultivars from South America, Australia, the United States, and the CIMMYT are known to possess the Sr24 resistance gene. On the basis of observed occurrence and postulated migration routes of the original Ug99 (race TTKSK), the confirmed presence of TTKST and PTKST in Eritrea increases the possibility for range expansion out of Africa by crossing the Red Sea and into the Arabian Peninsula. Future spread of TTKST and PTKST to western Asia is considered highly likely. References: (1) Y. Jin et al. Plant Dis. 92:923, 2008. (2) F. Mukoyi et al. Plant Dis. 95:1188, 2011. (3) Z. A. Pretorius et al. Plant Dis. 94:784, 2010. (4) R. P. Singh et al. Annu. Rev. Phytopathol. 49:465, 2011.

Chromosomal location and inheritance of stem rust resistance transferred from Hordeum bulbosum into cultivated barley (H. vulgare).

Stem rust, caused by Puccinia graminis f. sp. tritici, is an important disease on barley (Hordeum vulgare). Host resistance has effectively controlled stem rust, primarily through use of gene Rpg1. However, virulence to Rpg1 is present in North America, and a new race (TTKSK, or Ug99) from eastern Africa threatens barley production. A search for novel resistance was previously conducted, and an interspecific barley breeding line (212Y1) with introgressed chromatin from H. bulbosum was identified as carrying resistance to races MCCF and QCCJ. This study evaluated the inheritance of resistance in 212Y1 using populations from crosses to Morex (Rpg1 donor) and Q21861 (rpg4 donor) and the pathogen races MCCF (avirulent on Rpg1 and rpg4) and QCCJ (virulent on Rpg1 and avirulent on rpg4), and determined the chromosomal position of the introgression using genomic in situ hybridization (GISH) and chromosome-specific polymerase chain reaction (PCR)-based markers. Progeny from the 212Y1/Q21861 F(2) population segregated for resistant and susceptible plants, indicating different gene loci. Genetic analyses of Morex/212Y1 F(3) families fit a 7 homozygous resistant (HR):8 segregating:1 homozygous susceptible (HS) family segregation ratio to race MCCF, indicating that two genes controlled resistance. Plants in segregating families were in 3R:1S (Rpg1), 13R:3S (Rpg1+212Y1), and 1R:3S (212Y1) ratios. Genetic analyses of the same F(3) families fit a 1HR:2 segregating:1HS family segregation ratio to race QCCJ, indicating monogenic inheritance. Plants in segregating families were in a 1R: 3S ratio, indicating recessive inheritance in 212Y1. The introgression from H. bulbosum into H. vulgare was positioned on chromosome 6HS based on GISH and the PCR-based markers. No known stem rust resistance gene has previously been mapped to that region. Thus, it is proposed to name this novel gene from H. bulbosum as rpg6.

Detection of Virulence to Resistance Gene Sr36 Within the TTKS Race Lineage of Puccinia graminis f. sp. tritici.

The stem rust resistance gene Sr36 confers a near-immune resistance reaction to many races of Puccinia graminis f. sp. tritici and is highly effective against race TTKSK (syn. Ug99), which possesses unusually broad virulence combinations. Because this gene is widely used in United States soft winter wheat germplasm and cultivars, it has been considered to be an important source of resistance to TTKSK. In 2007, moderately susceptible infection responses were observed on wheat lines and cultivars carrying Sr36 in a field screening nursery for stem rust at Njoro, Kenya. We derived 18 single-pustule isolates from stem rust samples collected from the 2007 Njoro nursery. The isolates were evaluated for virulence on 20 North American stem rust differential lines and on wheat lines and cultivars carrying Sr36, Sr31+Sr36, and Sr24+Sr31. Of the 18 isolates, 10 produced infection types 3+ to 4 on line W2691SrTt-1 (monogenic for Sr36) and other lines that carry Sr36 and belonged to a new virulence phenotype that was not detected in previous years. These isolates were identified as race TTTSK. The remaining eight isolates were identified as races TTKSK (five isolates) and TTKST (three isolates), with avirulence and virulence, respectively, to Sr24. Thirteen simple sequence repeat (SSR) markers were used to examine the genetic relationships among the three races in the TTKS lineage. All isolates in the lineage shared an identical SSR genotype and were clearly different from North American races. In all, 16 wheat cultivars and 60 elite breeding lines, postulated to possess Sr36, were susceptible to race TTTSK. The occurrence of race TTTSK with combined virulence on Sr31 and Sr36 has further broadened the virulence spectrum of the TTKS lineage and rendered an important source of resistance ineffective.

Detection of Virulence to Resistance Gene Sr24 Within Race TTKS of Puccinia graminis f. sp. tritici.

The stem rust resistance gene Sr24 is effective against most races of Puccinia graminis f. sp. tritici, including race TTKS (syn. Ug99), and is used widely in commercial wheat cultivars worldwide. In 2006, susceptible infection responses were observed on wheat lines and cultivars carrying Sr24 in a field stem rust screening nursery at Njoro, Kenya. We derived 28 single-pustule isolates from stem rust samples collected from the 2006 Njoro nursery. The isolates were evaluated for virulence on 16 North American stem rust differential lines; on wheat lines carrying Sr24, Sr31, Sr38, and SrMcN; and on a wheat cultivar with a combination of Sr24 and Sr31. All isolates were identified as race TTKS with additional virulence on Sr31 and Sr38. These isolates were divided into two groups: group A (seven isolates and the two control isolates), producing a low infection type, and group B (21 isolates), producing a high infection type on Sr24, respectively. Isolates of group B represented a new variant of race TTKS with virulence to Sr24. Eighteen simple sequence repeat (SSR) markers were used to examine the genetic relationship between these two groups of isolates in race TTKS and five North American races (MCCF, QCCQ, RCRS, RTHS, and TPMK) that are representative of distinct lineage groups. All isolates of race TTKS shared an identical SSR genotype and were clearly different from North American races. The virulence and SSR data indicated that the new variant of race TTKS with Sr24 virulence likely has arisen via mutation within the TTKS genetic lineage. We propose to revise the North American stem rust nomenclature system by the addition of four genes (Sr24, Sr31, Sr38, and SrMcN) as the fifth set. This revision recognizes the virulence on Sr31 and differentiates isolates within race TTKS into two separate races: TTKSK and TTKST, with avirulence and virulence on Sr24, respectively. The occurrence of race TTKST with combined virulence on Sr24 and Sr31 has substantially increased the vulnerability of wheat to stem rust worldwide.

Letter Code System of Nomenclature for Puccinia graminis f. sp. avenae.

Current systems that describe the virulence phenotype in Puccinia graminis f. sp. avenae lack a systematic approach for the naming of races or to provide easily made comparisons of virulence among races. A new nomenclature system that simply and systematically characterizes virulence in P. graminis f. sp. avenae is described. The new system has the distinct advantage of providing easily seen relationships among races in contrast to previous nomenclature systems. This allows for easier interpretation of virulence relationships in the oat stem rust population and provides a large amount of virulence information with a minimum of written characters. This system uses single-gene differential lines with the resistance genes Pg1, Pg2, Pg3, Pg4, Pg6, Pg8, Pg9, Pg10, Pg12, Pg13, Pg15, and Pg16, grouped into three subsets of four lines in sequential Pg gene order. By grouping in sequential gene number order, the relationship of the new system to the "standard" system is easily seen. Each race is designated by a three-letter code, based on the seedling reaction (low or high) on 12 differential lines. The letter code nomenclature system is open ended and can be updated easily as new differential genes are identified. This system simply and precisely describes the virulence phenotypes of isolates of P. graminis f. sp. avenae, and allows for easily made comparisons of virulence of isolates collected over time and across geographical locations worldwide.

Characterization of Seedling Infection Types and Adult Plant Infection Responses of Monogenic Sr Gene Lines to Race TTKS of Puccinia graminis f. sp. tritici.

Stem rust, caused by Puccinia graminis f. sp. tritici, historically was one of the most destructive diseases of wheat and barley. The disease has been under effective control worldwide through the widespread use of host resistance. A number of stem rust resistance genes in wheat have been characterized for their reactions to specific races of P. graminis f. sp. tritici. Adult plant responses to race TTKS (also known as Ug99) of monogenic lines for Sr genes, a direct measurement of the effectiveness for a given gene, have not been investigated to any extent. This report summarizes adult plant infection responses and seedling infection types for monogenic lines of designated Sr genes challenged with race TTKS. High infection types at the seedling stage and susceptible infection responses in adult plants were observed on monogenic lines carrying Sr5, 6, 7a, 7b, 8a, 8b, 9a, 9b, 9d, 9g, 10, 11, 12, 15, 16, 17, 18, 19, 20, 23, 30, 31, 34, 38, and Wld-1. Monogenic lines of resistance genes Sr13, 22, 24, 25, 26, 27, 28, 32, 33, 35, 36, 37, 39, 40, 44, Tmp, and Tt-3 were effective against TTKS both at the seedling and adult plant stages. The low infection types to race TTKS observed for these resistance genes corresponded to the expected low infections of these genes to other incompatible races of P. graminis f. sp. tritici. The level of resistance conferred by these genes at the adult plant stage varied between highly resistant to moderately susceptible. The results from this study were inconclusive for determining the effectiveness of resistance genes Sr9e, 14, 21, and 29 against race TTKS. The understanding of the effectiveness of individual Sr genes against race TTKS will facilitate the utilization of these genes in breeding for stem rust resistance in wheat.

Evaluation of Avena spp. Accessions for Resistance to Oat Stem Rust.

Oat stem rust, caused by Puccinia graminis f. sp. avenae, can cause significant yield losses in the eastern prairie region of western Canada. Currently, the predominant race of P. graminis f. sp. avenae in this region is NA67. Few genes confer resistance to NA67, and none are present in any oat cultivars registered for production in Canada. To detect lines exhibiting resistance to race NA67, we evaluated 9,978 accessions from 22 Avena spp. in field nurseries from 2001 to 2004. In all, 35 accessions were highly resistant and 12 were moderately resistant, comprised mostly of the species A. strigosa. Seventy-one accessions had an intermediate response, comprised mostly of A. abyssinica, A. barbata, A. sterilis, and A. vaviloviana. All other accessions (9,860 = 98.8%) were susceptible to race NA67. Some highly resistant accessions were found to have been incorrectly classified previously as hexaploid species A. sativa or A. sterilis, and were confirmed by chromosome counts to be diploid or tetraploid. The most promising source of novel stem rust resistance is from the diploid species A. strigosa. Transfer of resistance from diploid and tetraploid species to A. sativa is very difficult, but the lines identified in this study should prove useful as new sources of resistance to oat stem rust.

Diversity and Sources of Multiple Disease Resistance in Hordeum spontaneum.

Hordeum spontaneum, the progenitor of cultivated barley, is known to be a rich source of disease resistance genes. The objective of this study was to assess the diversity of H. spontaneum accessions from Israel and Jordan for their reaction to six fungal pathogens of importance to cultivated barley in the United States and Canada. Overall, a high level of macro-scale (across collection sites) and micro-scale (within a collection site) diversity for disease reaction was found in the 116 accessions of H. spontaneum evaluated at the seedling stage. Additionally, genetic heterozygosity for resistance loci was common in H. spontaneum. The frequency of resistance in accessions from Jordan and Israel was high for Septoria speckled leaf blotch (77 and 98%, respectively), leaf rust (70 and 90%), net blotch (72 and 68%), and powdery mildew (58 and 70%); intermediate for spot blotch (53 and 46%); and low for stem rust (2 and 26%). The level of disease resistance in H. spontaneum was not strongly correlated with any of the weather variables (temperature, precipitation, and humidity) monitored near the collection sites. However, in general, resistance was more often found in germ plasm from mesic (e.g., Mediterranean coast) than in xeric (e.g., Negev Desert) areas. Two H. spontaneum accessions (Shechem 12-32 and Damon 11-11) were resistant to all six pathogens and may be useful parents in programs breeding barley for multiple disease resistance. The high level of diversity and heterozygosity for disease reaction found in this study indicates that H. spontaneum is an extraordinarily rich and largely untapped source of unique disease resistance alleles for cultivated barley improvement.

Fusarium Species Pathogenic to Barley and Their Associated Mycotoxins.

Epidemics of Fusarium head blight (FHB) occurred on barley in Minnesota, North Dakota, and South Dakota from 1993 to 1998. The Red River Valley region was most severely impacted by the disease based on assessments of FHB severity in grain samples harvested from commercial fields. Fusarium graminearum was the primary pathogen causing these FHB epidemics. It comprised from 62 to 64% of all Fusarium species isolated from infected kernels from 1994 to 1996. Fusarium poae (range of isolation 13 to 20%),F. sporotrichioides (10 to 17%), and F. avenaceum (6 to 10%) also were isolated from barley kernels and were likely involved in causing some FHB infection, but to a very limited extent. All four Fusarium species were pathogenic on barley in inoculation tests conducted in both the greenhouse and the field. Mycotoxin screens were performed on barley spikes inoculated with the respective species in the greenhouse. Spikes infected with F. graminearum contained deoxynivalenol and 15-acetyldeoxyni-valenol; those infected with F. sporotrichioides contained T-2 toxin, HT-2 toxin, and T-2 tetraol; and those infected with F. poae contained nivalenol. Some isolates of F. poae also produced 15-acetoxyscirpenol and scirpentriol. Although F. graminearum and DON are recognized as the primary FHB pathogen and mycotoxin, respectively, in barley, the possible presence of other Fusarium species and mycotoxins should not be overlooked.

Identification and mapping of leaf, stem and stripe rust resistance quantitative trait loci and their interactions in durum wheat.

Leaf rust (Puccinia triticina Eriks.), stripe rust (Puccinia striiformis f. tritici Eriks.) and stem rust (Puccinia graminis f. sp. tritici) cause major production losses in durum wheat (Triticum turgidum L. var. durum). The objective of this research was to identify and map leaf, stripe and stem rust resistance loci from the French cultivar Sachem and Canadian cultivar Strongfield. A doubled haploid population from Sachem/Strongfield and parents were phenotyped for seedling reaction to leaf rust races BBG/BN and BBG/BP and adult plant response was determined in three field rust nurseries near El Batan, Obregon and Toluca, Mexico. Stripe rust response was recorded in 2009 and 2011 nurseries near Toluca and near Njoro, Kenya in 2010. Response to stem rust was recorded in field nurseries near Njoro, Kenya, in 2010 and 2011. Sachem was resistant to leaf, stripe and stem rust. A major leaf rust quantitative trait locus (QTL) was identified on chromosome 7B at Xgwm146 in Sachem. In the same region on 7B, a stripe rust QTL was identified in Strongfield. Leaf and stripe rust QTL around DArT marker wPt3451 were identified on chromosome 1B. On chromosome 2B, a significant leaf rust QTL was detected conferred by Strongfield, and at the same QTL, a Yr gene derived from Sachem conferred resistance. Significant stem rust resistance QTL were detected on chromosome 4B. Consistent interactions among loci for resistance to each rust type across nurseries were detected, especially for leaf rust QTL on 7B. Sachem and Strongfield offer useful sources of rust resistance genes for durum rust breeding.

Rpr1, a gene required for Rpg1-dependent resistance to stem rust in barley.

Rpg1 is a stem rust resistance gene that has protected barley from severe losses for over 60 years in the US and Canada. It confers resistance to many, but not all, pathotypes of the stem rust fungus Puccinia graminis f. sp. tritici. A fast neutron induced deletion mutant, showing susceptibility to stem rust pathotype Pgt-MCC, was identified in barley cv. Morex, which carries Rpg1. Genetic and Rpg1 mRNA and protein expression level analyses showed that the mutation was a suppressor of Rpg1 and was designated Rpr1 (Required for P. graminis resistance). Genome-wide expression profiling, using the Affymetrix Barley1 GeneChip containing approximately 22,840 probe sets, was conducted with Morex and the rpr1 mutant. Of the genes represented on the Barley1 microarray, 20 were up-regulated and 33 were down-regulated by greater than twofold in the mutant, while the Rpg1 mRNA level remained constant. Among the highly down-regulated genes (greater than fourfold), genomic PCR, RT-PCR and Southern analyses identified that three genes (Contig4901_s_at, HU03D17U_s_at, and Contig7061_s_at), were deleted in the rpr1 mutant. These three genes mapped to chromosome 4(4H) bin 5 and co-segregated with the rpr1-mediated susceptible phenotype. The loss of resistance was presumed to be due to a mutation in one or more of these genes. However, the possibility exists that there are other genes within the deletions, which are not represented on the Barley1 GeneChip. The Rpr1 gene was not required for Rpg5- and rpg4-mediated stem rust resistance, indicating that it shows specificity to the Rpg1-mediated resistance pathway.