Development of wheat lines carrying stem rust resistance gene <Emphasis Type="Italic">Sr39</Emphasis> with reduced <Emphasis Type="Italic">Aegilops speltoides</Emphasis> chromatin and simple PCR markers for marker-assisted selection

The use of major resistance genes is a cost-effective strategy for preventing stem rust epidemics in wheat crops. The stem rust resistance gene Sr39 provides resistance to all currently known pathotypes of Puccinia graminis f. sp. tritici (Pgt) including Ug99 (TTKSK) and was introgressed together with leaf rust resistance gene Lr35 conferring adult plant resistance to P. triticina (Pt), into wheat from Aegilops speltoides. It has not been used extensively in wheat breeding because of the presumed but as yet undocumented negative agronomic effects associated with Ae. speltoides chromatin. This investigation reports the production of a set of recombinants with shortened Ae. speltoides segments through induction of homoeologous recombination between the wheat and the Ae. speltoides chromosome. Simple PCR-based DNA markers were developed for resistant and susceptible genotypes (Sr39#22r and Sr39#50s) and validated across a set of recombinant lines and wheat cultivars. These markers will facilitate the pyramiding of ameliorated sources of Sr39 with other stem rust resistance genes that are effective against the Pgt pathotype TTKSK and its variants.     

Genetics and mapping of seedling resistance to Ug99 stem rust in Canadian wheat cultivars ‘Peace’ and ‘AC Cadillac’

Stem rust (caused by Puccinia graminis Pers.:Pers. f. sp. tritici Eriks. & E. Henn.) has re-emerged as a threat to wheat production with the evolution of new pathogen races, namely TTKSK (Ug99) and its variants, in Africa. Deployment of resistant wheat cultivars has provided long-term control of stem rust. Identification of new resistance genes will contribute to future cultivars with broad resistance to stem rust. The related Canadian cultivars Peace and AC Cadillac show resistance to Ug99 at the seedling stage and in the field. The purpose of this study was to elucidate the inheritance and genetically map resistance to Ug99 in these two cultivars. Two populations were produced, an F_2:3 population from LMPG/AC Cadillac and a doubled haploid (DH) population from RL6071/Peace. Both populations showed segregation at the seedling stage for a single stem rust resistance (Sr) gene, temporarily named SrCad. SrCad was mapped to chromosome 6DS in both populations with microsatellite markers and a marker (FSD_RSA) that is tightly linked to the common bunt resistance gene Bt10. FSD_RSA was the closest marker to SrCad (≈1.6 cM). Evaluation of the RL6071/Peace DH population and a second DH population, AC Karma/87E03-S2B1, in Kenya showed that the combination of SrCad and leaf rust resistance gene Lr34 provided a high level of resistance to Ug99-type races in the field, whereas in the absence of Lr34 SrCad conferred moderate resistance. A survey confirmed that SrCad is the basis for all of the seedling resistance to Ug99 in Canadian wheat cultivars. While further study is needed to determine the relationship between SrCad and other Sr genes on chromosome 6DS, SrCad represents a valuable genetic resource for producing stem rust resistant wheat cultivars.     

Genetic mapping of stem rust resistance gene <Emphasis Type="Italic">Sr13</Emphasis> in tetraploid wheat (<Emphasis Type="Italic">Triticum turgidum</Emphasis> ssp. <Emphasis Type="Italic">durum</Emphasis> L.)

Wheat stem rust caused by Puccinia graminis f. sp. tritici, can cause significant yield losses. To combat the disease, breeders have deployed resistance genes both individually and in combinations to increase resistance durability. A new race, TTKSK (Ug99), identified in Uganda in 1999 is virulent on most of the resistance genes currently deployed, and is rapidly spreading to other regions of the world. It is therefore important to identify, map, and deploy resistance genes that are still effective against TTKSK. One of these resistance genes, Sr13, was previously assigned to the long arm of chromosome 6A, but its precise map location was not known. In this study, the genome location of Sr13 was determined in four tetraploid wheat (T. turgidum ssp. durum) mapping populations involving the TTKSK resistant varieties Kronos, Kofa, Medora and Sceptre. Our results showed that resistance was linked to common molecular markers in all four populations, suggesting that these durum lines carry the same resistance gene. Based on its chromosome location and infection types against different races of stem rust, this gene is postulated to be Sr13. Sr13 was mapped within a 1.2–2.8 cM interval (depending on the mapping population) between EST markers CD926040 and BE471213, which corresponds to a 285-kb region in rice chromosome 2, and a 3.1-Mb region in Brachypodium chromosome 3. These maps will be the foundation for developing high-density maps, identifying diagnostic markers, and positional cloning of Sr13.     

Characterization of Sr9h,a wheat stem rust resistance allele effective to Ug99

Key message

Wheat stem rust resistance gene SrWeb is an allele at the Sr9 locus that confers resistance to Ug99.

Abstract

Race TTKSK (Ug99) of Puccinia graminis f. sp. tritici, the causal fungus of stem rust, threatens global wheat production because of its broad virulence to current wheat cultivars. A recently identified Ug99 resistance gene from cultivar Webster, temporarily designated as SrWeb, mapped near the stem rust resistance gene locus Sr9. We determined that SrWeb is also present in Ug99 resistant cultivar Gabo 56 by comparative mapping and an allelism test. Analysis of resistance in a population segregating for both Sr9e and SrWeb demonstrated that SrWeb is an allele at the Sr9 locus, which subsequently was designated as Sr9h. Webster and Gabo 56 were susceptible to the Ug99-related race TTKSF+ from South Africa. Race TTKSF+ possesses unique virulence to uncharacterized Ug99 resistance in cultivar Matlabas. This result validated that resistance to Ug99 in Webster and Gabo 56 is conferred by the same gene: Sr9h. The emergence of pathogen virulence to several resistance genes that are effective to the original Ug99 race TTKSK, including Sr9h, suggests that resistance genes should be used in combinations in order to increase resistance durability.     

QTL mapping of resistance to race Ug99 of Puccinia graminis f. sp. tritici in durum wheat (Triticum durum Desf.)

Stem rust caused by Puccinia graminis f. sp. tritici was historically one of the most destructive diseases of wheat worldwide. The evolution and rapid migration of race TTKSK (Ug99) and derivatives, first detected in Uganda in 1999, are of international concern due to the virulence of these races to widely used stem rust resistance genes. In attempts to identify quantitative trait loci (QTL) linked with resistance to stem rust race Ug99, 95 recombinant inbred lines that were developed from a cross between two durum wheat varieties, Kristal and Sebatel, were evaluated for reaction to stem rust. Seven field trials at two locations were carried out in main and off seasons. In addition to the natural infection, the nursery was also artificially inoculated with urediniospores of stem rust race Ug99 and a mixture of locally collected stem rust urediniospores. A genetic map was constructed based on 207 simple sequence repeat (SSR) and two sequence tagged site loci. Using composite interval mapping, nine QTL for resistance to stem rust were identified on chromosomes 1AL, 2AS, 3BS, 4BL, 5BL, 6AL 7A, 7AL and 7BL. These results suggest that durum wheat resistance to stem rust is oligogenic and that there is potential to identify previously uncharacterized resistance genes with minor effects. The SSR markers that are closely linked to the QTL can be used for marker-assisted selection for stem rust resistance in durum wheat.     

A consensus map for Ug99 stem rust resistance loci in wheat

Key message

This consensus map of stem rust genes, QTLs, and molecular markers will facilitate the identification of new resistance genes and provide a resource of in formation for development of new markers for breeding wheat varieties resistant to Ug99.

Abstract

The global effort to identify new sources of resistance to wheat stem rust, caused by Puccinia graminis f. sp. tritici race group Ug99 has resulted in numerous studies reporting both qualitative genes and quantitative trait loci. The purpose of our study was to assemble all available information on loci associated with stem rust resistance from 21 recent studies on Triticum aestivum L. (bread wheat) and Triticum turgidum subsp. durum desf. (durum wheat). The software LPmerge was used to construct a stem rust resistance loci consensus wheat map with 1,433 markers incorporating Single Nucleotide Polymorphism, Diversity Arrays Technology, Genotyping-by-Sequencing as well as Simple Sequence Repeat marker information. Most of the markers associated with stem rust resistance have been identified in more than one population. Several loci identified in these populations map to the same regions with known Sr genes including Sr2, SrND643, Sr25 and Sr57 (Lr34/Yr18/Pm38), while other significant markers were located in chromosome regions where no Sr genes have been previously reported. This consensus map provides a comprehensive source of information on 141 stem rust resistance loci conferring resistance to stem rust Ug99 as well as linked markers for use in marker-assisted selection.     

Identification and validation of SSR markers linked to the stem rust resistance gene Sr6 on the short arm of chromosome 2D in wheat

The wheat stem rust resistance gene Sr6, present in several wheat cultivars, confers a high level of resistance against a wide range of races of Puccinia graminis f. sp. tritici. Resistance conferred by Sr6 is influenced by temperature, light intensity, and genetic background of the recipient genotype. Here, we report the identification and validation of molecular markers linked to Sr6 that can be used for the detection of this gene in wheat breeding programs. A mapping population of 136 F₂ plants and their F_2:3 families derived from a cross between near-isogenic lines, ‘Chinese Spring’ and ISr6-Ra, were screened for stem rust reaction in the seedling stage. Bulked segregant analysis (BSA) based on seedling tests was used to screen 418 SSR markers that covered the entire genome of wheat. Four markers, Xwmc453, Xcfd43, Xcfd77, and Xgwm484, were mapped within a chromosome region that spanned 9.7 cM from Sr6. The closest markers, Xwmc453 and Xcfd43, were linked to Sr6 at a distance of 1.1 and 1.5 cM, respectively. The markers Xwmc453 and Xcfd43 amplified Sr6-specific marker alleles that were diagnostic for Sr6 in a diverse set of 46 wheat accessions and breeding lines developed and/or collected in Australia, Canada, China, Egypt, Ethiopia, Kenya, Mexico, South Africa, and USA. These markers can now be used for marker-assisted selection of Sr6 and for pyramiding it with other stem rust resistance genes.     

The application of DNA marker and doubled-haploid technology for stacking multiple stem rust resistance genes in wheat

In wheat, the use of gene “pyramids” or “stacks” of major genes that confer resistance to all local strains of the fungal stem rust pathogen Puccinia graminis f. sp. tritici (Pgt) can increase durability of resistance where wheat cultivars with the single gene components are not widely deployed. Stacking two or more resistance genes becomes a breeding challenge, particularly when pathogen races that discriminate the genes are not available. The use of DNA markers and doubled-haploid technology provides a route for producing lines homozygous for multiple resistance genes. We have applied this approach to produce gene pyramids of two or more of the stem rust resistance genes Sr24 and new sources of SrR, Sr31 and Sr26 on reduced alien chromatin in the genetic backgrounds of Westonia and Pavon wheat. These genes, which are all derived from “alien” sources (SrR and Sr31 from rye, Sr24 and Sr26 from Agropyron elongatum) each provide resistance to all currently known pathotypes of Pgt in Australia, and SrR and Sr26 also provide resistance against all the variants of stem rust race Ug99 (TTKS group).     

Searching for novel sources of field resistance to Ug99 and Ethiopian stem rust races in durum wheat via association mapping

Puccinia graminis f. sp. tritici, the causative agent of stem rust in wheat, is a devastating disease of durum wheat. While more than 50 stem rust resistance (Sr) loci have been identified in wheat, only a few of them have remained effective against Ug99 (TTKSK race) and other durum-specific Ethiopian races. An association mapping (AM) approach based on 183 diverse durum wheat accessions was utilized to identify resistance loci for stem rust response in Ethiopia over four field-evaluation seasons and artificial inoculation with Ug99 and a mixture of durum-specific races. The panel was profiled with simple sequence repeat, Diversity Arrays Technology and sequence-tagged site markers (1,253 in total). The resistance turned out to be oligogenic, with twelve QTL-tagging markers that were significant (P < 0.05) across three or four seasons. R ² values ranged from 1.1 to 11.3 %.Twenty-four additional single-marker/QTL regions were found to be significant over two seasons. The AM results confirmed the role of Sr13, previously described in bi-parental mapping studies, and the role of chromosome regions putatively harbouring Sr9, Sr14, Sr17 and Sr28. Three minor QTLs were coincident with those reported in hexaploid wheat and five overlapped with those recently reported in the Sebatel × Kristal durum mapping population. Thirteen single-marker/QTL regions were located in chromosome regions where no Sr genes/QTLs have been previously reported. The allelic variation identified in this study is readily available and can be exploited for marker-assisted selection, thus providing additional opportunities for a more durable stem rust resistance under field conditions.     

Development and characterization of a compensating wheat-Thinopyrum intermedium Robertsonian translocation with Sr44 resistance to stem rust (Ug99)

The emergence of the highly virulent Ug99 race complex of the stem rust fungus (Puccinia graminis Pers. f. sp. tritici Eriks. and Henn.) threatens wheat (Triticum aestivum L.) production worldwide. One of the effective genes against the Ug99 race complex is Sr44, which was derived from Thinopyrum intermedium (Host) Barkworth and D.R. Dewey and mapped to the short arm of 7J (designated 7J#1S) present in the noncompensating T7DS-7J#1L?7J#1S translocation. Noncompensating wheat-alien translocations are known to cause genomic duplications and deficiencies leading to poor agronomic performance, precluding their direct use in wheat improvement. The present study was initiated to produce compensating wheat-Th. intermedium Robertsonian translocations with Sr44 resistance. One compensating RobT was identified consisting of the wheat 7DL arm translocated to the Th. intermedium 7J#1S arm resulting in T7DL?7J#1S. The T7DL?7J#1S stock was designated as TA5657. The 7DL?7J#1S stock carries Sr44 and has resistance to the Ug99 race complex. This compensating RobT with Sr44 resistance may be useful in wheat improvement. In addition, we identified an unnamed stem rust resistance gene located on the 7J#1L arm that confers resistance not only to Ug99, but also to race TRTTF, which is virulent to Sr44. However, the action of the second gene can be modified by the presence of suppressors in the recipient wheat cultivars.     

A novel Robertsonian translocation event leads to transfer of a stem rust resistance gene (<Emphasis Type="Italic">Sr52</Emphasis>) effective against race Ug99 from <Emphasis Type="Italic">Dasypyrum</Emphasis><Emphasis Type="Italic">villosum</Emphasis> into bread wheat

Stem rust (Puccinia graminis f. sp. tritici Eriks. & E. Henn.) (the causal agent of wheat stem rust) race Ug99 (also designated TTKSK) and its derivatives have defeated several important stem rust resistance genes widely used in wheat (Triticum aestivum L.) production, rendering much of the worldwide wheat acreage susceptible. In order to identify new resistance sources, a large collection of wheat relatives and genetic stocks maintained at the Wheat Genetic and Genomic Resources Center was screened. The results revealed that most accessions of the diploid relative Dasypyrum villosum (L.) Candargy were highly resistant. The screening of a set of wheat–D. villosum chromosome addition lines revealed that the wheat–D. villosum disomic addition line DA6V#3 was moderately resistant to race Ug99. The objective of the present study was to produce and characterize compensating wheat–D. villosum whole arm Robertsonian translocations (RobTs) involving chromosomes 6D of wheat and 6V#3 of D. villosum through the mechanism of centric breakage-fusion. Seven 6V#3-specific EST–STS markers were developed for screening F₂ progeny derived from plants double-monosomic for chromosomes 6D and 6V#3. Surprisingly, although 6D was the target chromosome, all recovered RobTs involved chromosome 6A implying a novel mechanism for the origin of RobTs. Homozygous translocations (T6AS·6V#3L and T6AL·6V#3S) with good plant vigor and full fertility were selected from F₃ families. A stem rust resistance gene was mapped to the long arm 6V#3L in T6AS·6V#3L and was designated as Sr52. Sr52 is temperature-sensitive and is most effective at 16°C, partially effective at 24°C, and ineffective at 28°C. The T6AS·6V#3L stock is a new source of resistance to Ug99, is cytogenetically stable, and may be useful in wheat improvement.     

Diagnostic and co-dominant PCR markers for wheat stem rust resistance genes Sr25 and Sr26

Wheat stem rust, caused by Puccinia graminis f. sp. tritici, is one of the most destructive diseases of wheat. A new race of the pathogen named TTKSK (syn. Ug99) and its derivatives detected in East Africa are virulent to many designated and undesignated stem rust resistance genes. The emergence and spread of those races pose an imminent threat to wheat production worldwide. Genes Sr25 and Sr26 transferred into wheat from Thinopyrum ponticum are effective against these new races. DNA markers for Sr25 and Sr26 are needed to pyramid both genes into adapted germplasm. The previously published dominant markers Gb for Sr25 and Sr26#43 for Sr26 were validated with eight wheat lines with or without Sr25 or Sr26. We tested six published STS (sequence tagged site) markers amplifying diagnostic bands of Th. ponticum. Marker BF145935 consistently amplified well and can be used as a co-dominant marker for Sr25. Among 16 STS markers developed from wheat ESTs mapped to deletion bin 6AL8-0.90-1.00, none was co-dominant for tagging Sr26. However, five 6A-specific markers were identified. Multiplex PCR with marker Sr26#43 and 6A-specific marker BE518379 can be used as a co-dominant marker for Sr26. The co-dominant markers for Sr25 and Sr26 were validated with 37 lines with known stem rust resistance genes. A diverse set of germplasm consisting 170 lines from CIMMYT, China, USA and other counties were screened with the co-dominant markers for Sr25 and Sr26. Five lines with the diagnostic fragment for Sr25 were identified, and they all have ‘Wheatear’ in their pedigrees, which is known to carry Sr25. None of the 170 lines tested had Sr26, as expected.     

Inheritance of resistance to Ug99 stem rust in wheat cultivar Norin 40 and genetic mapping of Sr42

Stem rust, caused by Puccinia graminis f. sp. tritici, is a devastating disease of wheat. The emergence of race TTKSK (Ug99) and new variants in Africa threatens wheat production worldwide. The best method of controlling stem rust is to deploy effective resistance genes in wheat cultivars. Few stem rust resistance (Sr) genes derived from the primary gene pool of wheat confer resistance to TTKSK. Norin 40, which carries Sr42, is resistant to TTKSK and variants TTKST and TTTSK. The goal of this study was to elucidate the inheritance of resistance to Ug99 in Norin 40 and map the Sr gene(s). A doubled haploid (DH) population of LMPG-6/Norin 40 was evaluated for resistance to the race TTKST. Segregation of 248 DH lines fitted a 1:1 ratio (χ (2) 1:1= 0.58, p = 0.45), indicating a single gene in Norin 40 conditioned resistance to Ug99. This was confirmed by an independent F(2:3) population also derived from the cross LMPG-6/Norin 40 where a 1:2:1 ratio (χ (2)1:2:1 = 0.69, p = 0.71) was observed following the inoculation with race TTKSK. Mapping with DNA markers located this gene to chromosome 6DS, the known location of Sr42. PCR marker FSD_RSA co-segregated with Sr42, and simple sequence repeat (SSR) marker BARC183 was closely linked (0.5 cM) to Sr42. A previous study found close linkage between FSD_RSA and SrCad, a temporarily designated gene that also confers resistance to Ug99, thus Sr42 may be the same gene or allelic. Marker FSD_RSA is suitable for marker-assisted selection (MAS) in wheat breeding programs to improve stem rust resistance, including Ug99.     

Fine genetic mapping fails to dissociate durable stem rust resistance gene Sr2 from pseudo-black chaff in common wheat (Triticum aestivum L.)

The broad-spectrum stem rust resistance gene Sr2 has provided protection in wheat against Puccinia graminis Pers. f. sp. tritici for over 80 years. The Sr2 gene and an associated dark pigmentation trait, pseudo-black chaff (PBC), have previously been localized to the short arm of chromosome 3B. In a first step towards the positional-based cloning of Sr2, we constructed a high-resolution map of this region. The wheat EST (wEST) deletion bin mapping project provided tightly linked cDNA markers. The rice genome sequence was used to infer the putative gene order for orthologous wheat genes and provide additional markers once the syntenic interval in rice was identified. We used this approach to map six wESTs that were collinear with the physical order of the corresponding genes on rice chromosome 1 suggesting there are no major re-arrangements between wheat and rice in this region. We were unable to separate by recombination the tightly linked morphological trait, PBC from the stem rust resistance gene suggesting that either a single gene or two tightly linked genes control both traits.     

Introgression of stem rust resistance genes SrTA10187 and SrTA10171 from Aegilops tauschii to wheat

Aegilops tauschii, the diploid progenitor of the wheat D genome, is a readily accessible germplasm pool for wheat breeding as genes can be transferred to elite wheat cultivars through direct hybridization followed by backcrossing. Gene transfer and genetic mapping can be integrated by developing mapping populations during backcrossing. Using direct crossing, two genes for resistance to the African stem rust fungus race TTKSK (Ug99), were transferred from the Ae. tauschii accessions TA10187 and TA10171 to an elite hard winter wheat line, KS05HW14. BC₂ mapping populations were created concurrently with developing advanced backcross lines carrying rust resistance. Bulked segregant analysis on the BC₂ populations identified marker loci on 6DS and 7DS linked to stem rust resistance genes transferred from TA10187 and TA10171, respectively. Linkage maps were developed for both genes and closely linked markers reported in this study will be useful for selection and pyramiding with other Ug99-effective stem rust resistance genes. The Ae. tauschii-derived resistance genes were temporarily designated SrTA10187 and SrTA10171 and will serve as valuable resources for stem rust resistance breeding.     

Development and characterization of wheat-<Emphasis Type="Italic">Ae. searsii</Emphasis> Robertsonian translocations and a recombinant chromosome conferring resistance to stem rust

The emergence of a new highly virulent race of stem rust (Puccinia graminis tritici), Ug99, rapid evolution of new Ug99 derivative races overcoming resistance of widely deployed genes, and spread towards important wheat growing areas now potentially threaten world food security. Exploiting novel genes effective against Ug99 from wild relatives of wheat is one of the most promising strategies for the protection of the wheat crop. A new source of resistance to Ug99 was identified in the short arm of the Aegilops searsii chromosome 3S^s by screening wheat- Ae. searsii introgression libraries available as individual chromosome and chromosome arm additions to the wheat genome. For transferring this resistance gene into common wheat, we produced three double-monosomic chromosome populations (3A/3S^s, 3B/3S^s and 3D/3S^s) and then applied integrated stem rust screening, molecular maker analysis, and cytogenetic analysis to identify resistant wheat-Ae. searsii Robertsonian translocation. Three Robertsonian translocations (T3AL·3S^sS, T3BL·3S^sS and T3DL·3S^sS) and one recombinant (T3DS-3S^sS·3S^sL) with stem rust resistance were identified and confirmed to be genetically compensating on the basis of genomic in situ hybridization, analysis of 3A, 3B, 3D and 3S^sS-specific SSR/STS-PCR markers, and C-banding. In addition, nine SSR/STS-PCR markers of 3S^sS-specific were developed for marker-assisted selection of the resistant gene. Efforts to reduce potential linkage drag associated with 3S^sS of Ae. searsii are currently under way.     

Protecting South Asian Wheat Production from Stem Rust (Ug99) Epidemic

The Ug99 group of stem rust races (Puccinia graminis Pers. f. sp. tritici Eriks. & E. Henn.) has evolved and migrated. While the original variant overcame the widely deployed gene Sr31, and Sr21 (in Chinese Spring background), but not Sr21 in Einkorn, a new strain of Ug99, virulent on Sr24, was detected in 2006 and caused a severe epidemic in 2007 in Kenya. Forms virulent on Sr36 and Sr21 were identified in 2007. Likewise, an Ug99 variant virulent to both Sr21 and Sr24 was identified in 2008 in Kenya. Simultaneously, the original strain spread to Yemen and Sudan in 2006. Fears of a spread into Asia were confirmed when this race was detected in Iran in 2007. This has raised serious concerns that Ug99 could follow the same migratory route from Africa to Asia as Yr9 and cause major epidemics across the epidemiological region of South Asia. In 2005–06, screening in Kenya and Ethiopia of wheat materials from Asian countries revealed a very low frequency of lines resistant to Ug99 and its variants. Under the umbrella of the Borlaug Global Rust Initiative (BGRI), significant efforts have been made to counter the challenges posed by Ug99 and its derivative races. Diverse sources of resistance to the pathogen have been identified and incorporated in high‐yielding wheat backgrounds. The most promising strategy has been to deploy spring wheat varieties possessing adult plant resistance (APR) in infested and bordering areas to decrease inoculum amounts and slow down the development of new virulence, for example four CIMMYT genotypes with Sr2+ have been released in Afghanistan and their seed is also distributed in region bordering Iran. For an immediate remedy, race‐specific resistance genes can be deployed in combinations using marker‐assisted selection. Several Ug99‐resistant varieties have already been released in South Asian countries (Afghanistan, India, Nepal, Bangladesh and Pakistan), and seed dissemination is underway. The Ug99 risk in the region can be reduced to minimum levels by identifying, releasing and providing seed of high‐yielding and resistant cultivars.     

Association mapping and gene-gene interaction for stem rust resistance in CIMMYT spring wheat germplasm

The recent emergence of wheat stem rust Ug99 and evolution of new races within the lineage threatens global wheat production because they overcome widely deployed stem rust resistance (Sr) genes that had been effective for many years. To identify loci conferring adult plant resistance to races of Ug99 in wheat, we employed an association mapping approach for 276 current spring wheat breeding lines from the International Maize and Wheat Improvement Center (CIMMYT). Breeding lines were genotyped with Diversity Array Technology (DArT) and microsatellite markers. Phenotypic data was collected on these lines for stem rust race Ug99 resistance at the adult plant stage in the stem rust resistance screening nursery in Njoro, Kenya in seasons 2008, 2009 and 2010. Fifteen marker loci were found to be significantly associated with stem rust resistance. Several markers appeared to be linked to known Sr genes, while other significant markers were located in chromosome regions where no Sr genes have been previously reported. Most of these new loci colocalized with QTLs identified recently in different biparental populations. Using the same data and Q?+?K covariate matrices, we investigated the interactions among marker loci using linear regression models to calculate P values for pairwise marker interactions. Resistance marker loci including the Sr2 locus on 3BS and the wPt1859 locus on 7DL had significant interaction effects with other loci in the same chromosome arm and with markers on chromosome 6B. Other resistance marker loci had significant pairwise interactions with markers on different chromosomes. Based on these results, we propose that a complex network of gene-gene interactions is, in part, responsible for resistance to Ug99. Further investigation may provide insight for understanding mechanisms that contribute to this resistance gene network.     

Haplotype diversity of stem rust resistance loci in uncharacterized wheat lines

Stem rust is one of the most destructive diseases of wheat worldwide. The recent emergence of wheat stem rust race Ug99 (TTKS based on the North American stem rust race nomenclature system) and related strains threaten global wheat production because they overcome widely used genes that had been effective for many years. Host resistance is likely to be more durable when several stem rust resistance genes are pyramided in a single wheat variety; however, little is known about the resistance genotypes of widely used wheat germplasm. In this study, a diverse collection of wheat germplasm was haplotyped for stem rust resistance genes Sr2, Sr22, Sr24, Sr25, Sr26, Sr36, Sr40, and 1A.1R using linked microsatellite or simple sequence repeat (SSR) and sequence tagged site (STS) markers. Haplotype analysis indicated that 83 out of 115 current wheat breeding lines from the International Maize and Wheat Improvement Center (CIMMYT) likely carry Sr2. Among those, five out of 94 CIMMYT spring lines tested had both Sr2 and Sr25 haplotypes. Five out of 22 Agriculture Research Service (ARS) lines likely have Sr2 and a few have Sr24, Sr36, and 1A.1R. Two out of 43 Chinese accessions have Sr2. No line was found to have the Sr26 and Sr40 haplotypes in this panel of accessions. DArT genotyping was used to identify new markers associated with the major stem resistance genes. Four DArT markers were significantly associated with Sr2 and one with Sr25. Principal component analysis grouped wheat lines from similar origins. Almost all CIMMYT spring wheats were clustered together as a large group and separated from the winter wheats. The results provide useful information for stem rust resistance breeding and pyramiding.     

Identification of Ug99 stem rust resistance loci in winter wheat germplasm using genome-wide association analysis

The evolution of a new race of stem rust, generally referred to as Ug99, threatens global wheat production because it can overcome widely deployed resistance genes that had been effective for many years. To identify loci conferring resistance to Ug99 in wheat, a genome-wide association study was conducted using 232 winter wheat breeding lines from the International Winter Wheat Improvement Program. Breeding lines were genotyped with diversity array technology, simple sequence repeat and sequence-tagged site markers, and phenotyped at the adult plant stage for resistance to stem rust in the stem rust resistance screening nursery at Njoro, Kenya during 2009-2011. A mixed linear model was used for detecting marker-trait associations. Twelve loci associated with Ug99 resistance were identified including markers linked to known genes Sr2 and Lr34. Other markers were located in the chromosome regions where no Sr genes have been previously reported, including one each on chromosomes 1A, 2B, 4A and 7B, two on chromosome 5B and four on chromosome 6B. The same data were used for investigating epistatic interactions between markers with or without main effects. The marker csSr2 linked to Sr2 interacted with wPt4930 on 6BS and wPt729773 in an unknown location. Another marker, csLV34 linked to Lr34, also interacted with wPt4930 on 6BS and wPt4916 on 2BS. The frequent involvement of wPt4916 on 2BS and wPt4930 on 6BS in interactions with other significant loci on the same or different chromosomes suggested complex genetic control for adult plant resistance to Ug99 in winter wheat germplasm.