Pablo Olivera

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.

Emergence of Virulence to SrTmp in the Ug99 Race Group of Wheat Stem Rust, Puccinia graminis f. sp. tritici, in Africa

detected at two locations in Uganda (Rubaya and Muko in Kabale region) and at five locations in Rwanda (Kinigi, Rwerere, Rufungo, Gatebe, and Kamenyo). Three isolates derived from stem rust samples collected on cv. PBW343 (carrying Sr31) in Sakha in the Nile Delta region in Egypt were also typed as TTKTK. In addition, DNA from isolates of race TTKTK were analyzed using a diagnostic qPCR assay (Ug99 RG stage-1, Szabo, unpublished data), which confirmed that these samples belong to the Ug99 lineage. The identification of SrTmp virulence in the Ug99 race group in several countries in one year emphasizes the relevance of coordinated international surveillance efforts and utilization of diverse sources of resistance to control stem rust in wheat. Further studies are in progress to determine the detailed relationship of the newly emerged races and other Pgt isolates identified in the Ug99 group.

Resistance of Sharon goatgrass (Aegilops sharonensis) to fungal diseases of wheat

Sharon goatgrass (Aegilops sharonensis) is a wild relative of wheat that is native to Israel and Lebanon. The importance of A. sharonensis as a source of new resistance genes for wheat warrants additional research on the characterization of accessions for economically important genes. Thus, the objectives of this study were to evaluate a collection of A. sharonensis accessions for resistance to seven important fungal diseases of wheat and assess the phenotypic diversity of the germplasm for disease reaction. The frequency of resistance in A. sharonensis was highest to powdery mildew (79 to 83%) and leaf rust (60 to 77%). Resistance to stem rust also was common, although the percentage of resistant accessions varied markedly depending on the pathogen race-from 13% to race TTTT to 72% to race QCCJ. The frequency of resistance was intermediate to stripe rust (45%) and low to tan spot (15 to 29%) and spot blotch (0 to 34%). None of the A. sharonensis accessions was resistant to Fusarium head blight. Many of the accessions tested exhibited heterogeneous reactions (i.e., had both resistant and susceptible plants) to one or more of the diseases, suggesting that heterozygosity may be present at some resistance loci. Substantial variation was observed in the level of diversity to individual diseases because Shannons Equitability index ranged from 0.116 (for Fusarium head blight) to 0.994 (for tan spot). A high level of diversity was found both between and within collection sites. Moreover, differences in the geographic distribution of resistant accessions were observed. For example, accessions from northern Israel generally were less diverse and less resistant to leaf rust and stripe rust than accessions from more southern locations. Four A. sharonensis accessions were highly resistant to most of the diseases evaluated and may provide a source of unique resistance genes for introgression into cultivated wheat.

Kenyan Isolates of Puccinia graminis f. sp. tritici from 2008 to 2014: Virulence to SrTmp in the Ug99 Race Group and Implications for Breeding Programs

Frequent emergence of new variants in the Puccinia graminis f. sp. tritici Ug99 race group in Kenya has made pathogen survey a priority. We analyzed 140 isolates from 78 P. graminis f. sp. tritici samples collected in Kenya between 2008 and 2014 and identified six races, including three not detected prior to 2013. Genotypic analysis of 20 isolates from 2013 and 2014 collections showed that the new races TTHST, TTKTK, and TTKTT belong to the Ug99 race group. International advanced breeding lines were evaluated against an isolate of TTKTT (Sr31, Sr24, and SrTmp virulence) at the seedling stage. From 169 advanced lines from Kenya, 23% of lines with resistance to races TTKSK and TTKST were susceptible to TTKTT and, from two North American regional nurseries, 44 and 91% of resistant lines were susceptible. Three lines with combined resistance genes were developed to facilitate pathogen monitoring and race identification. These results indicate the increasing virulence and variability in the Kenyan P. graminis f. sp. tritici population and reveal vulnerabilities of elite germplasm to new races.

Association Mapping Reveals Novel Stem Rust Resistance Loci in Durum Wheat at the Seedling Stage

Wheat stem rust rapidly evolves new virulence to resistance genes. Recently emerged races in East Africa, such as TTKSK (or Ug99), possess broad virulence to durum cultivars, and only a limited number of genes provide resistance. An association mapping (AM) study conducted on 183 durum wheat accessions has allowed us to identify 41 quantitative trait loci (QTLs; determination coefficient [R2] values from 1.1 to 23.1%) for seedling resistance to one or more of four highly virulent stem rust races: TRTTF, TTTTF, TTKSK (Ug99), and JRCQC, two of which (TRTTF and JRCQC) were isolated from Ethiopia. Among these loci, 24 are novel, while the remaining 17 overlapped with loci previously shown to provide field resistance in Ethiopia and/or chromosome regions known to harbor designated stem rust resistance designated loci (Sr). The identified loci were either effective against multiple races or race specific, particularly for race JRCQC. Our results highlight that stem rust resistance in durum wheat is governed in part by loci for resistance across multiple races, and in part by race‐specific ones (23 and 18, respectively). Collectively, these results provide useful information to improve the effectiveness of marker‐assisted selection towards the release of durum wheat cultivars with durable stem rust resistance.

First Report of the Ug99 Race Group of Wheat Stem Rust, Puccinia graminis f. sp. tritici, in Egypt in 2014

American stem rust differential lines following standard race-typing procedure and infection type (IT) criteria determining virulence and avirulence (Jin et al. 2008). In addition, three supplemental tester lines of Siouxland (carrying Sr24+Sr31), Sisson (carrying Sr31+Sr36), and Triumph 64 (donor of SrTmp) were included to confirm virulence/avirulence to Sr24, Sr31, Sr36, and SrTmp. The experiments were repeated two to three times. Three races in the Ug99 race group were detected; TTKST (four isolates, IT 3+4 for Sr24, Sr31, and cv. Siouxland) from Al-Sharqia, TTKTK (13 isolates, IT 4 for Sr31, SrTmp, and cv. Triumph 64) from Sakha, and TTKSK (2 isolates, IT 4 for Sr31) from Nubaria. This is the first confirmation of races in the Ug99 race group in Egypt, thereby extending the geographical distribution of Ug99-related races. Since Egypt may play a role as green-bridge for P. graminis f. sp. tritici between East and North African countries and the wheat belts in the Middle East and Mediterranean regions, the rust surveillance efforts should be intensified in affected countries as well as in neighboring regions.

Genetics of resistance to wheat leaf rust, stem rust, and powdery mildew in Aegilops sharonensis

Aegilops sharonensis (Sharon goatgrass) is a wild relative of wheat and a rich source of genetic diversity for disease resistance. The objectives of this study were to determine the genetic basis of leaf rust, stem rust, and powdery mildew resistance in A. sharonensis and also the allelic relationships between genes controlling resistance to each disease. Progeny from crosses between resistant and susceptible accessions were evaluated for their disease reaction at the seedling and/or adult plant stage to determine the number and action of genes conferring resistance. Two different genes conferring resistance to leaf rust races THBJ and BBBB were identified in accessions 1644 and 603. For stem rust, the same single gene was found to confer resistance to race TTTT in accessions 1644 and 2229. Resistance to stem rust race TPMK was conferred by two genes in accessions 1644 and 603. A contingency test revealed no association between genes conferring resistance to leaf rust race THBJ and stem rust race TTTT or between genes conferring resistance to stem rust race TTTT and powdery mildew isolate UM06-01, indicating that the respective resistance genes are not linked. Three accessions (1644, 2229, and 1193) were found to carry a single gene for resistance to powdery mildew. Allelism tests revealed that the resistance gene in accession 1644 is different from the respective single genes present in either 2229 or 1193. The simple inheritance of leaf rust, stem rust, and powdery mildew resistance in A. sharonensis should simplify the transfer of resistance to wheat in wide crosses.

Identification of Resistance to Races of Puccinia graminis f. sp. tritici with Broad Virulence in Triticale (×Triticosecale)

Triticale (×Triticosecale), an amphiploid of wheat (mainly Triticum turgidum) and cereal rye (Secale cereale), is an excellent source of resistance to wheat stem rust, caused by Puccinia graminis f. sp. tritici. A collection of 567 triticale accessions originating from 21 countries was evaluated at the seedling stage for reaction to races of P. graminis f. sp. tritici with broad virulence, including TTKSK, TRTTF, and TTTTF. A high frequency (78.4%) of accessions was resistant to race TTKSK, with low infection types ranging from 0; to X. A selection of 353 TTKSK-resistant accessions was evaluated for reaction to three South African isolates of P. graminis f. sp. tritici with single and/or combined virulences to stem rust resistance genes SrSatu, Sr27, and SrKw present in triticale. Genes SrSatu, Sr27, and SrKw were postulated to be present in 141 accessions and contributed to TTKSK resistance. The remaining 212 resistant accessions may possess uncharacterized genes or combinations of known genes that could not be determined with these isolates. These accessions were further evaluated for resistance to races TTKST, TPMKC, RKQQC, RCRSC, QTHJC, QCCSM, and MCCFC. Resistance remained effective across the entire set of races in the majority of the accessions (n = 200), suggesting that the resistances are effective against a broad spectrum of virulence. In all, 129 (79.6%) resistant accessions with noncharacterized genes were resistant to moderately resistant in field stem rust nurseries at Debre Zeit (Ethiopia) and St. Paul (Minnesota). Results from evaluating F2 populations derived from resistant-susceptible crosses revealed that resistance to TTKSK in triticale was conferred mostly by single genes with dominant effects.

Genotype-by-sequencing facilitates genetic mapping of a stem rust resistance locus in Aegilops umbellulata, a wild relative of cultivated wheat

BackgroundWild relatives of wheat play a significant role in wheat improvement as a source of genetic diversity. Stem rust disease of wheat causes significant yield losses at the global level and stem rust pathogen race TTKSK (Ug99) is virulent to most previously deployed resistance genes. Therefore, the objective of this study was to identify loci conferring resistance to stem rust pathogen races including Ug99 in an Aegilops umbelluata bi-parental mapping population using genotype-by-sequencing (GBS) SNP markers.ResultsA bi-parental F2:3 population derived from a cross made between stem rust resistant accession PI 298905 and stem rust susceptible accession PI 542369 was used for this study. F2 individuals were evaluated with stem rust race TTTTF followed by testing F2:3 families with races TTTTF and TTKSK. The segregation pattern of resistance to both stem rust races suggested the presence of one resistance gene. A genetic linkage map, comprised 1,933 SNP markers, was created for all seven chromosomes of Ae. umbellulata using GBS. A major stem rust resistance QTL that explained 80% and 52% of the phenotypic variations for TTTTF and TTKSK, respectively, was detected on chromosome 2U of Ae. umbellulata.ConclusionThe novel resistance gene for stem rust identified in this study can be transferred to commercial wheat varieties assisted by the tightly linked markers identified here. These markers identified through our mapping approach can be a useful strategy to identify and track the resistance gene in marker-assisted breeding in wheat.

Discovery and characterization of two new stem rust resistance genes in Aegilops sharonensis

Key messageWe identified two novel wheat stem rust resistance genes, Sr-1644-1Sh and Sr-1644-5Sh in Aegilops sharonensis that are effective against widely virulent African races of the wheat stem rust pathogen.AbstractStem rust is one of the most important diseases of wheat in the world. When single stem rust resistance (Sr) genes are deployed in wheat, they are often rapidly overcome by the pathogen. To this end, we initiated a search for novel sources of resistance in diverse wheat relatives and identified the wild goatgrass species Aegilops sharonesis (Sharon goatgrass) as a rich reservoir of resistance to wheat stem rust. The objectives of this study were to discover and map novel Sr genes in Ae. sharonensis and to explore the possibility of identifying new Sr genes by genome-wide association study (GWAS). We developed two biparental populations between resistant and susceptible accessions of Ae. sharonensis and performed QTL and linkage analysis. In an F6 recombinant inbred line and an F2 population, two genes were identified that mapped to the short arm of chromosome 1Ssh, designated as Sr-1644-1Sh, and the long arm of chromosome 5Ssh, designated as Sr-1644-5Sh. The gene Sr-1644-1Sh confers a high level of resistance to race TTKSK (a member of the Ug99 race group), while the gene Sr-1644-5Sh conditions strong resistance to TRTTF, another widely virulent race found in Yemen. Additionally, GWAS was conducted on 125 diverse Ae. sharonensis accessions for stem rust resistance. The gene Sr-1644-1Sh was detected by GWAS, while Sr-1644-5Sh was not detected, indicating that the effectiveness of GWAS might be affected by marker density, population structure, low allele frequency and other factors.