We identify the wheat stem rust resistance gene Sr50 (using physical mapping, mutation and complementation) as homologous to barley Mla, encoding a coiled-coil nucleotide-binding leucine-rich repeat (CC-NB-LRR) protein. We show that Sr50 confers a unique resistance specificity different from Sr31 and other genes on rye chromosome 1RS, and is effective against the broadly virulent Ug99 race lineage. Extensive haplotype diversity at the rye Sr50 locus holds promise for mining effective resistance genes.
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In much of the world, resistance to stem rust in wheat, caused by Puccinia graminis f. sp. tritici, is based at least in part on the gene Sr31. During February 1999, high levels of stem rust infection were observed on entries in wheat (Triticum aestivum) grown in a nursery at Kalengyere Research Station in Uganda. Because several of the rusted entries were known to carry the 1BL-1RS chromosome translocation containing the Sr31, Lr26, and Yr9 genes for rust resistance, virulence to Sr31 was suspected. Urediniospores, collected in bulk from rusted stems of seven entries containing Sr31, were suspended in light mineral oil and sprayed on primary leaves of 7-day-old seedlings of South African wheat cv. Gamtoos (=Veery #3, pedigree: Kvz/Buho‘S’//Kal/BB). Plants were kept overnight at 19 to 21°C in a dew chamber before placement in a greenhouse at 18 to 25°C. After ≈14 days, urediniospores were collected from large, susceptible-type stem rust pustules and subsequently increased on Gamtoos, which served as a selective host for the new rust culture, designated Pgt-Ug99. Pathogenicity of Pgt-Ug99 was studied in seedling tests of available wheats containing Sr31, as well as other stem rust differential lines. All seedling tests were conducted at least three times in independent inoculations. Isolate Pgt-Ug99 was not virulent to Avocet‘S’/Yr9 (Australian line containing Sr26) or Oom Charl (South African cultivar) but was virulent to the other Sr31 testers: Alondra ‘S’, Bobwhite, Chokka, Clement, Federation/Kavkaz, Gamtoos, Grebe, Kavkaz, Letaba, Line E/Kavkaz, RL6078, and Veery ‘S’. Virulence to Sr31 (infection types [ITs] 3-3 to 3++4) was clearly contrasted by the low reactions (ITs 0; to 1) produced by UVPgt53, a South African pathotype avirulent to Sr31. According to the reactions of the differential lines, Pgt-Ug99 is avirulent to Sr21, -22, -24, -25, -26, -27, -29, -32, -33, -34, -35, -36, -39, -40, -42, and -43, Agi, and Em and virulent to Sr5, -6, -7b, -8a, -8b, -9b, -9e, -9g, -11, -15, -17, -30, -31, and -38. Virulence to the T. ventricosum-derived gene Sr38, which is linked to Lr37 and Yr17 and occurs in cultivars from Australia, the United Kingdom, and the United States, was not known previously (1). Both Pgt-Ug99 and UVPgt53 produced a continuum of ITs (; to 2+3) on Petkus rye (obtained from the USDA-ARS National Small Grains Collection, Aberdeen, ID), the original Sr31 donor source. Pgt-Ug99 did not appear more virulent than UVPgt53 on Petkus. All triticales tested, as well as oat cv. Overberg, were highly resistant to Pgt-Ug99. According to McIntosh et al. (1), Huerta-Espino mentioned a Sr31-virulent culture from Turkey, but this could not be confirmed. Should the Sr31-virulent pathotype migrate out of Uganda, it poses a major threat to wheat production in countries where the leading cultivars have resistance based on this gene.
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.
Aegilops tauschii Coss., the D genome donor of hexaploid wheat, Triticum aestivum L., has been used extensively for the transfer of agronomically important traits to wheat, including stem rust resistance genes Sr33, Sr45, and Sr46. To identify potentially new stem rust resistance genes in A. tauschii germplasm, we evaluated 456 nonduplicated accessions deposited in the USDA National Small Grains Collection (Aberdeen, ID) and the Wheat Genetic and Genomic Resources Center collection (Kansas State University, Manhattan, KS), with races TTKSK (Ug99), TRTTF, TTTTF, TPMKC, RKQQC, and QTHJC of Puccinia graminis Pers.:Pers. f. sp. tritici Eriks. & E. Henn. Ninety-eight accessions (22%) were identified as resistant to race TTKSK. A broad range of resistant infection types (; to 2+) were found in reaction to race TTKSK. Resistance was significantly associated among most of the races in pairwise comparisons. However, resistance was largely race specific. Only 12 of the accessions resistant to race TTKSK were also resistant to the other five races. Results from this germplasm screening will facilitate further studies on the genetic characterization of accessions with potentially novel sources of resistance to race TTKSK.
Stem rust resistance gene Sr45, discovered in Aegilops tauschii, the progenitor of the D -genome of wheat, is effective against commercially important Puccinia graminis f. sp. tritici races prevalent in Australia, South Africa and the Ug99 race group. A synthetic hexaploid wheat (RL5406) generated by crossing Ae. tauschii accession RL5289 (carrying Sr45 and the leaf rust resistance gene Lr21) with a tetraploid experimental line ‘TetraCanthatch’ was previously used as the source in the transfer of these rust resistance genes to other hexaploid cultivars. Previous genetic studies on hexaploid wheats mapped Sr45 on the short arm of chromosome 1D with the following gene order: centromere–Sr45–Sr33–Lr21–telomere. To identify closely linked markers, we fine mapped the Sr45 region in a large mapping population generated by crossing CS1D5406 (disomic substitution line with chromosome 1D of RL5406 substituted for Chinese Spring 1D) with Chinese Spring. Closely linked markers based on 1DS-specific microsatellites, expressed sequence tags and AFLP were useful in the delineation of the Sr45 region. Sequences from an AFLP marker amplified a fragment that was linked with Sr45 at a distance of 0.39 cM. The fragment was located in a bacterial artificial chromosome clone of contig (ctg)2981 of the Ae. tauschii accession AL8/78 physical map. A PCR marker derived from clone MI221O11 of ctg2981 amplified 1DS-specific sequence that harboured an 18-bp indel polymorphism that specifically tagged the Sr45 carrying haplotype. This new Sr45 marker can be combined with a previously reported marker for Lr21, which will facilitate selecting Sr45 and Lr21 in breeding populations.
As there are numerous pathogen species that cause disease and limit yields of crops, such as wheat (Triticum aestivum), single genes that provide resistance to multiple pathogens are valuable in crop improvement. The mechanistic basis of multi-pathogen resistance is largely unknown. Here we use comparative genomics, mutagenesis and transformation to isolate the wheat Lr67 gene, which confers partial resistance to all three wheat rust pathogen species and powdery mildew. The Lr67 resistance gene encodes a predicted hexose transporter (LR67res) that differs from the susceptible form of the same protein (LR67sus) by two amino acids that are conserved in orthologous hexose transporters. Sugar uptake assays show that LR67sus, and related proteins encoded by homeoalleles, function as high-affinity glucose transporters. LR67res exerts a dominant-negative effect through heterodimerization with these functional transporters to reduce glucose uptake. Alterations in hexose transport in infected leaves may explain its ability to reduce the growth of multiple biotrophic pathogen species.
The stem rust fungus, particularly race TTKSK (Ug99), poses a serious threat to world wheat production. Gene Sr42 or SrCad (which could be the same gene or an allele of Sr42) is effective against race TTKSK. However, known genetic markers for Sr42 are mostly SSR markers which are generally labor intensive to use. In this study, we mapped a race TTKSK resistance gene derived from PI 595667 at the same locus as Sr42 on chromosome 6DS. Based on position, pedigree and infection-type information, we propose that this gene is SrCad (Sr42). We enriched the genetic map for the Sr42 region using genotyping by sequencing (GBS) and array-derived SNP markers. In total, 21 SNP markers were discovered, spanning a genetic distance of 27.2 cM. Nine of them are derived from GBS and twelve from the Illumina iSelect 90K SNP assay. Ten of the twenty-one SNP markers are closely linked (<2.2 cM, or co-segregating) with Sr42. We converted five of the closely linked SNP markers into uniplex KASP assays which will better facilitate marker-assisted selection. We validated the KASP assay in a doubled haploid wheat population derived from a three-way cross between accessions PI 410954, RB07, and Faller that shared an uncharacterized resistance gene mapped at approximately the same locus as PI 595667. The development of closely linked (co-segregating), codominant, sequence-based SNP assays will aid marker-assisted selection and map-based cloning of Sr42.
The transfer of alien genes to crop plants using chromosome engineering has been attempted infrequently in tetraploid durum wheat (Triticum turgidum L. subsp. durum). Here, we report a highly efficient approach for the transfer of two genes conferring resistance to stem rust race Pgt-TTKSK (Ug99) from goatgrass (Aegilops speltoides) to tetraploid wheat. The durum line DAS15, carrying the stem rust resistance gene Sr47 derived from Ae. speltoides, was crossed, and backcrossed, to durum 5D(5B) aneuploids to induce homeologous pairing. After a final cross to ‘Rusty’ durum, allosyndetic recombinants were recovered. The Ae. speltoides chromosomal segment carrying Sr47 was found to have two stem rust resistance genes. One gene conditioning an infection type (IT) 2 was located in the same chromosomal region of 2BS as Sr39 and was assigned the temporary gene symbol SrAes7t. Based on ITs observed on a diverse set of rust races, SrAes7t may be the same as Sr39. The second gene conditioned an IT 0; and was located on chromosome arm 2BL. This gene retained the symbol Sr47 because it had a different IT and map location from other stem rust resistance genes derived from Ae. speltoides. Allosyndetic recombinant lines carrying each gene on minimal alien chromosomal segments were identified as were molecular markers distinguishing each alien segment. This study demonstrated that chromosome engineering of Ae. speltoides segments is feasible in tetraploid wheat. The Sr47 gene confers high-level and broad spectrum resistance to stem rust and should be very useful in efforts to control TTKSK.
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.
Widely virulent races of the stem rust pathogen (Puccinia graminis f. sp. tritici) such as those isolated from Africa (e.g., TTKSK, isolate synonym Ug99) threaten wheat production worldwide. To identify Aegilops accessions with effective resistance to such virulent stem rust races, up to 10 different species from Israel were evaluated against African races TTKSK, TTKST, and TTTSK and the Israeli race TTTTC as seedlings in the greenhouse. A wide diversity of stem rust reactions was observed across the Aegilops spp. and ranged from highly resistant (i.e., infection type 0) to highly susceptible (infection type 4). The frequency of resistance within a species to races TTTTC and TTKSK ranged from 7 and 14%, respectively, in Aegilops searsii to 98 and 100% in AE. speltoides. In all, 346 accessions were found resistant to the three African races and 138 accessions were resistant (or heterogeneous with a resistant component) to all four races. The species with broadly resistant accessions included Ae. longissima (59 accessions), Ae. peregrina (47 accessions), Ae. sharonensis (15 accessions), Ae. geniculata (9 accessions), Ae. kotschyi (5 accessions), and Ae. bicornis (3 accessions). Few geographical trends or correlations with climatic variables were observed with respect to stem rust resistance in the Aegilops spp. The exception was Ae. longissima infected with race TTTTC, where a high frequency of resistance was found in central and northern Israel and a very low frequency in southern Israel (Negev desert region). This geographical trend followed a pattern of annual precipitation in Israel, and a significant correlation was found between this variable and resistance in Ae. longissima. Although difficult, it is feasible to transfer resistance genes from Aegilops spp. into wheat through conventional wide-crossing schemes or, alternatively, a cloning and transformation approach. The broadly resistant accessions identified in this study will be valuable in these research programs.