Association analysis of stem rust resistance in U.S. winter wheat

Citation: Zhang D, Bowden RL, Yu J, Carver BF, Bai G (2014) Association Analysis of Stem Rust Resistance in U.S. Winter Wheat. PLoS ONE 9(7): e103747. https://doi.org/10.1371/journal.pone.0103747Stem rust has become a renewed threat to global wheat production after the emergence and spread of race TTKSK (also known as Ug99) and related races from Africa. To elucidate U.S. winter wheat resistance genes to stem rust, association mapping was conducted using a panel of 137 lines from cooperative U.S. winter wheat nurseries from 2008 and simple sequence repeat (SSR) and sequence tagged site (STS) markers across the wheat genome. Seedling infection types were evaluated in a greenhouse experiment using six U.S. stem rust races (QFCSC, QTHJC, RCRSC, RKQQC, TPMKC and TTTTF) and TTKSK, and adult plant responses to bulked U.S. races were evaluated in a field experiment. A linearization algorithm was used to convert the qualitative Stakman scale seedling infection types for quantitative analysis. Association mapping successfully detected six known stem rust seedling resistance genes in U.S. winter wheat lines with frequencies: Sr6 (12%), Sr24 (9%), Sr31 (15%), Sr36 (9%), Sr38 (19%), and Sr1RS[superscript Amigo] (8%). Adult plant resistance gene Sr2 was present in 4% of lines. SrTmp was postulated to be present in several hard winter wheat lines, but the frequency could not be accurately determined. Sr38 was the most prevalent Sr gene in both hard and soft winter wheat and was the most effective Sr gene in the adult plant field test. Resistance to TTKSK was associated with nine markers on chromosome 2B that were in linkage disequilibrium and all of the resistance was attributed to the Triticum timopheevii chromosome segment carrying Sr36. Potential novel rust resistance alleles were associated with markers Xwmc326-203 on 3BL, Xgwm160-195 and Xwmc313-225 on 4AL near Sr7, Xgwm495-182 on 4BL, Xwmc622-147 and Xgwm624-146 on 4DL, and Xgwm334-123 on 6AS near Sr8. Xwmc326-203 was associated with adult plant resistance to bulked U.S. races and Xgwm495-182 was associated with seedling resistance to TTKSK

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

Stem 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 goat grass species Aegilops sharonesis (Sharon goatgrass) as a substantial 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 (one of the Ug99 lineage races), 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

Association mapping of stem rust race TTKSK resistance in US barley breeding germplasm

KEY MESSAGE: Loci conferring resistance to the highly virulent African stem rust race TTKSK were identified in advanced barley breeding germplasm and positioned to chromosomes 5H and 7H using an association mapping approach. ABSTRACT: African races of the stem rust pathogen (Puccinia graminis f. sp. tritici) are a serious threat to barley production worldwide because of their wide virulence. To discover and characterize resistance to African stem rust race TTKSK in US barley breeding germplasm, over 3,000 lines/cultivars were assessed for resistance at the seedling stage in the greenhouse and also the adult plant stage in the field in Kenya. Only 12 (0.3 %) and 64 (2.1 %) lines exhibited a resistance level comparable to the resistant control at the seedling and adult plant stage, respectively. To map quantitative trait loci (QTL) for resistance to race TTKSK, an association mapping approach was conducted, utilizing 3,072 single nucleotide polymorphism (SNP) markers. At the seedling stage, two neighboring SNP markers (0.8 cM apart) on chromosome 7H (11_21491 and 12_30528) were found significantly associated with resistance. The most significant one found was 12_30528; thus, the resistance QTL was named Rpg-qtl-7H-12_30528. At the adult plant stage, two SNP markers on chromosome 5H (11_11355 and 12_31427) were found significantly associated with resistance. This resistance QTL was named Rpg-qtl-5H-11_11355 for the most significant marker identified. Adult plant resistance is of paramount importance for stem rust. The marker associated with Rpg-qtl-5H-11_11355 for adult plant resistance explained only a small portion of the phenotypic variation (0.02); however, this QTL reduced disease severity up to 55.0 % under low disease pressure and up to 21.1 % under heavy disease pressure. SNP marker 11_11355 will be valuable for marker-assisted selection of adult plant stem rust resistance in barley breeding. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s00122-014-2297-8) contains supplementary material, which is available to authorized users

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

Quantitative and Qualitative Stem Rust Resistance Factors in Barley Are Associated with Transcriptional Suppression of Defense Regulons

Stem 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

Resistance to TTKSK and TTTSK Races of Puccinia graminis f.sp. tritici in Ethiopian Tetraploid Wheat Accessions

Ethiopia is the leading wheat producing country in East Africa. East Africa is a known hotspot for the evolution of new rust races of wheat. A case in point could be the evolution of a new race of stem rust called Ug99 that overcame the resistance deployed in wheat breeding for stem rust for many decades. Tetraploid wheat is an excellent source of stem rust resistance. In Ethiopia recent race analysis works showed that races TTKSK and TTTSK are the two dominant races distributed in major wheat producing areas of the country such as Arsi and Bale. Taking this into account, searching for sources of stem rust resistance to races TTKSK and TTTSK is an important breeding strategy. Therefore, this study was conducted to identify sources of resistance to the two major stem rust races from tetraploid wheat accessions. A total of 40 tetraploid wheat accessions comprising Triticcum durum, T. diccoccum and T. turgidum were evaluated both at the seedling stage at Kulumssa and adult plant growth stages at Arsi Robe under natural epidemics. In the seedling stage, most of the accessions were resistant to both races. The infection type range from ; to 2+. 2+ is the maximum infection type that was displayed by few of the accessions. All the tetraploid accessions displayed low infection types to both races. Stem rust occurred at epidemic level at Arsi Robe compared to other testing locations. Almost half of the accessions were completely susceptible to stem rust infection in the field condition. Five accessions exhibited a completely immune reaction to the prevailing races. The rest displayed a trace level of moderately susceptible reaction. Those accession that combined seedling and adult plant resistance to stem rust infection could be utilized in the durum wheat breeding program of Ethiopia.   Keywords: Accessions, infection type, tetraploid, race, TTKSK, TTTSK

Mining a Collection of triticum diccoccoid Landraces for Resistance to Races of Puccinia graminis f. sp. tritici at Seedling Stage

Stem rust caused by Puccinia graminis Pers. f. sp. tritici Eriks. and E. Henn. (Pgt), is one of the most destructive pathogen of wheat which causing  considerable yield losses. It has become a renewed threat to global wheat production after the emergence and spread of new race known as TTKSK or  Ug99 and related races from Africa. Races of the pathogen in the “Ug99 lineage” are of international concern due to their virulence for widely used stem  rust resistance genes and their spread throughout Africa. Seedlings of 183 diccocciod wheat accessions which were assembled by University of Bologna,  department of agricultural sciences were evaluated for their response to stem rust (Puccinia graminis f. sp. tritici) infection under greenhouse condition  at University of Minnesota St. Paul campus during 2012 with an objective of identifying diccoccoid wheat accessions that could serve as sources of  resistance to stem rust to enhance durable resistant variety development. Seedling infection types were evaluated using three stem rust races viz TTKSK,  TRTTF and TTTTF. A high level of phenotypic variation was observed in response to races TTKSK, TRTTF and TTTTF in the test entries, allowing for selection  in these germplasm as a pre-breeding work. Out of the tested accessions, 32 diccoccoid wheat accessions exhibited low infection types (0–2)  response to all the three races and hence selected as a source of resistance to stem rust. These wheat germplasm which are identified as resistant along  with adult plant resistant germplasm will be promising genetic stocks for accumulating resistances genes to acquire durable resistance and long lasting  variety/ies.&nbsp

Inheritance and mapping of stem rust resistance of wheat line PI 410966

Stem rust caused by Puccinia graminis f. sp tritici of wheat (Triticum aestivum L.) is one of the most destructive cereal diseases globally. Concern about the disease has increased since 1999 with the discovery in Uganda of a new virulent race of Pgt, designated as race TTKSK (also known as Ug99). The objectives of this experiment were to characterize the resistance and to determine the chromosomal location of the stem rust resistance in the spring wheat line PI 410966. A mapping population was developed from a cross between PI 410966 and a susceptible wheat line OK3040. An inoculation test with isolate 04KEN156/04 of race TTKSK was conducted at the USDA-ARS Cereal Disease Laboratory in the F6:7 generation, and the F6:7 phenotypic data were used to genetically map the resistance gene to the centromeric region on chromosome 2BS. The single locus explained the observed F6:7 resistant and susceptible scores. The location of the gene and molecular marker banding profiles of the diagnostic markers suggest that the stem rust resistance gene in PI 410966 could be a new gene, an allele of Sr36, or Sr36

Developing adapted wheat lines with broad-spectrum resistance to stem rust: Introgression of Sr59 through backcrossing and selections based on genotyping-by-sequencing data

Control of stem rust, caused by Puccinia graminis f.sp. tritici, a highly destructive fungal disease of wheat, faces continuous challenges from emergence of new virulent races across wheat-growing continents. Using combinations of broad-spectrum resistance genes could impart durable stem rust resistance. This study attempted transfer of Sr59 resistance gene from line TA5094 (developed through CSph1bM-induced T2DS center dot 2RL Robertsonian translocation conferring broad-spectrum resistance). Poor agronomic performance of line TA5094 necessitates Sr59 transfer to adapted genetic backgrounds and utility evaluations for wheat improvement. Based on combined stem rust seedling and molecular analyses, 2070 BC1F1 and 1230 BC2F1 plants were derived from backcrossing BAJ#1, KACHU#1, and REEDLING#1 with TA5094. Genotyping-by-sequencing (GBS) results revealed the physical positions of 15,116 SNPs on chromosome 2R. The adapted genotypes used for backcrossing were found not to possess broad-spectrum resistance to selected stem rust races, whereas Sr59-containing line TA5094 showed resistance to all races tested. Stem rust seedling assays combined with kompetitive allele-specific PCR (KASP) marker analysis successfully selected and generated the BC2F2 population, which contained the Sr59 gene, as confirmed by GBS. Early-generation data from backcrossing suggested deviations from the 3:1 segregation, suggesting that multiple genes may contribute to Sr59 resistance reactions. Using GBS marker data (40,584 SNPs in wheat chromosomes) to transfer the recurrent parent background to later-generation populations resulted in average genome recovery of 71.2% in BAJ#1*2/TA5094, 69.8% in KACHU#1*2/TA5094, and 70.5% in REEDLING#1*2/TA5094 populations. GBS data verified stable Sr59 introgression in BC2F2 populations, as evidenced by presence of the Ph1 locus and absence of the 50,936,209 bp deletion in CSph1bM. Combining phenotypic selections, stem rust seedling assays, KASP markers, and GBS data substantially accelerated transfer of broad-spectrum resistance into adapted genotypes. Thus, this study demonstrated that the Sr59 resistance gene can be introduced into elite genetic backgrounds to mitigate stem rust-related yield losses