Validation and identification of molecular markers linked to the leaf rust resistance gene <Emphasis Type="Italic">Lr</Emphasis>28 in wheat

Leaf rust resistance gene Lr28 controls one of the important resistances in the Indian subcontinent against the most prevalent Puccinia triticina pathotype 77-5. Pyramiding Lr28 with other resistance genes would therefore, provide durable resistance against rust, a process that can be facilitated by DNA markers. A microsatellite marker wmc313 linked to Lr28 at a distance of 5.0 cM was identified in the population HD2329 × HW2037. The marker was validated in another population developed from WL711× CS + Lr28: 2D/2M 3/8 (acc. 2956) as well as in a few near- isogenic lines (NILs) carrying gene Lr28. Compared to the previously reported marker TPSCAR SCS421₅₇₀, wmc313 is more closely linked to Lr28. Both these markers flanking the gene should be useful in the deployment of Lr28 into the breeding program using marker-assisted selection allowing pyramiding with other effective genes to confer durable resistance.     

Fine mapping of LrSV2, a race-specific adult plant leaf rust resistance gene on wheat chromosome 3BS

Key message

Fine mapping permits the precise positioning of genes within chromosomes, prerequisite for positional cloning that will allow its rational use and the study of the underlying molecular action mechanism.

Abstract

Three leaf rust resistance genes were identified in the durable leaf rust resistant Argentinean wheat variety Sinvalocho MA: the seedling resistance gene Lr3 on distal 6BL and two adult plant resistance genes, LrSV1 and LrSV2, on chromosomes 2DS and 3BS, respectively. To develop a high-resolution genetic map for LrSV2, 10 markers were genotyped on 343 F2 individuals from a cross between Sinvalocho MA and Gama6. The closest co-dominant markers on both sides of the gene (3 microsatellites and 2 STMs) were analyzed on 965 additional F2s from the same cross. Microsatellite marker cfb5010 cosegregated with LrSV2 whereas flanking markers were found at 1 cM distal and at 0.3 cM proximal to the gene. SSR markers designed from the sequences of cv Chinese Spring BAC clones spanning the LrSV2 genetic interval were tested on the recombinants, allowing the identification of microsatellite swm13 at 0.15 cM distal to LrSV2. This delimited an interval of 0.45 cM around the gene flanked by the SSR markers swm13 and gwm533 at the subtelomeric end of chromosome 3BS.     

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.     

Genetics and mapping of stem rust resistance to Ug99 in the wheat cultivar Webster

New races of wheat stem rust, namely TTKSK (Ug99) and its variants, pose a threat to wheat production in the regions where they are found. The accession of the wheat cultivar Webster (RL6201) maintained at the Cereal Research Centre in Winnipeg, Canada, shows resistance to TTKSK and other races of stem rust. The purpose of this study was to study the inheritance of seedling resistance to stem rust in RL6201 and genetically map the resistance genes using microsatellite (SSR) markers. A population was produced by crossing the stem rust susceptible line RL6071 with Webster. The F₂ and F₃ were tested with TPMK, a stem rust race native to North America. The F₃ was also tested with TTKSK. Two independently assorting genes were identified in RL6201. Resistance to TPMK was conferred by Sr30, which was mapped with microsatellites on chromosome 5DL. The second gene, temporarily designated SrWeb, conferred resistance to TTKSK. SrWeb was mapped to chromosome 2BL using SSR markers. Comparison with previous genetic maps showed that SrWeb occupies a locus near Sr9. Further analysis will be required to determine if SrWeb is a new gene or an allele of a previously identified gene.     

Development of COS-SNP and HRM markers for high-throughput and reliable haplotype-based detection of Lr14a in durum wheat (Triticum durum Desf.)

Leaf rust (Puccinia triticina Eriks. & Henn.) is a major disease affecting durum wheat production. The Lr14a-resistant gene present in the durum wheat cv. Creso and its derivative cv. Colosseo is one of the best characterized leaf-rust resistance sources deployed in durum wheat breeding. Lr14a has been mapped close to the simple sequence repeat markers gwm146, gwm344 and wmc10 in the distal portion of the chromosome arm 7BL, a gene-dense region. The objectives of this study were: (1) to enrich the Lr14a region with single nucleotide polymorphisms (SNPs) and high-resolution melting (HRM)-based markers developed from conserved ortholog set (COS) genes and from sequenced Diversity Array Technology (DArT^®) markers; (2) to further investigate the gene content and colinearity of this region with the Brachypodium and rice genomes. Ten new COS-SNP and five HRM markers were mapped within an 8.0 cM interval spanning Lr14a. Two HRM markers pinpointed the locus in an interval of <1.0 cM and eight COS-SNPs were mapped 2.1–4.1 cM distal to Lr14a. Each marker was tested for its capacity to predict the state of Lr14a alleles (in particular, Lr14-Creso associated to resistance) in a panel of durum wheat elite germplasm including 164 accessions. Two of the most informative markers were converted into KASPar^® markers. Single assay markers ubw14 and wPt-4038-HRM designed for agarose gel electrophoresis/KASPar^® assays and high-resolution melting analysis, respectively, as well as the double-marker combinations ubw14/ubw18, ubw14/ubw35 and wPt-4038-HRM–ubw35 will be useful for germplasm haplotyping and for molecular-assisted breeding.     

Sequence tagged microsatellites for the Xgwm533 locus provide new diagnostic markers to select for the presence of stem rust resistance gene Sr2 in bread wheat ( Triticum aestivum L.)

The stem rust resistance gene Sr2 has provided durable broad-spectrum, adult-plant resistance to the fungal pathogen Puccinia graminis Pers. f. sp. tritici throughout wheat-growing regions of the world for more than 50 years. The ability to select for Sr2 in wheat breeding programs was recently improved by the identification of a tightly linked microsatellite marker gwm533. This marker typically amplifies a 120-bp polymerase chain reaction fragment from wheat lines carrying Sr2. In instances where the 120-bp fragment is not associated with the presence of Sr2, DNA sequence analysis has shown that a second allele was amplified, differing in the structure of the microsatellite repeat. To discriminate this allelic homoplasy (alleles identical in size, but not identical by descent), sequence-tagged microsatellites (STM) markers were developed for the Xgwm533 locus. These markers were shown to be diagnostic for the presence of Sr2 in a wide range of germplasm, representative of all major wheat varieties historically grown in Australia. The STMs will be particularly useful for marker-assisted selection in Southern Australian breeding programs, where the use of the marker gwm533 is often precluded by the presence of the non-Sr2-associated 120-bp allele in the pedigree of current breeding germplasm. The STMs also revealed a high incidence of previously undetected allelic homoplasy at the Xgwm533 locus and may have broader utility in genetic research and breeding, as this locus is also reported to be strongly associated with a major gene conferring resistance to Fusarium head blight.     

Microsatellite Marker Linked to a Leaf Rust Resistance Gene from Triticum monococcum L Transferred to Bread Wheat

Triticum monococcum L, a diploid wheat species closely related to the A genome of cultivated wheats, is highly resistant to leaf rust. A synthetic amphiploid, T. monococcum — T. durum was crossed with T. aestivum cv WL711, highly susceptible to leaf rust. Leaf rust resistant derivatives were selected among backcross generations with the recurrent parent WL711 and cytologically analysed. Chromosome number of the leaf rust resistant BC₁F₃ progenies varied from 39 to 44. Six leaf rust resistant and susceptible bulks from different BC₁F₃ progenies were analysed using 29 wheat microsatellite(WMS) markers already mapped on A genome of bread wheat and found polymorphic among parents. One T. monococcum specific allele of WMS gwm136 locus was found to be closely linked to the leaf rust resistance gene in all the resistant bulks. Differential chromosome number, frequency of univalents and multivalents, however, indicated that the critical T. monococcum chromosome might be present in addition to the A genome chromosomes of wheat, substituted either for the B or D genome chromosome of wheat or translocated to chromosome 1A of wheat in one or the other bulks. The association of the T. monococcum specific allele of WMS gwm136 locus to leaf rust resistance was further confirmed from bulked segregant analysis in BC₂F₁ generation.     

Genetic analysis and molecular mapping of a stripe rust resistance gene derived from Psathynrostachys huashanica Keng in wheat line H9014-121-5-5-9

Stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), is one of the most devastating diseases worldwide and is also an important disease in China. The wheat translocation line H9014-121-5-5-9 was originally developed from interspecific hybridization between wheat (Triticum aestivum L.) line 7182 and Psathyrostachys huashanica Keng. This translocation line showed resistance to predominant stripe rust races in China when it was tested with nine races of Pst. To determine the inheritance and map the resistance gene, segregating populations were developed from the cross between H9014-121-5-5-9 and the susceptible cultivar Mingxian 169. The seedlings of the F1, F2, and F2:3 generations were tested with race CYR31. The results showed that the resistance in H9014-121-5-5-9 was conferred by a single dominant gene. Bulked segregant analysis and simple sequence repeat (SSR) markers were used to identify polymorphic markers associated with the resistance gene locus. Seven polymorphic SSR markers were linked to the resistance gene. A linkage map was constructed according to the genotypes of the seven SSR markers and the resistance gene. Based on the SSR marker positions on the wheat chromosome, the resistance gene was assigned on chromosome 1AL, temporarily designated YrHA. Based on chromosomal location, reaction patterns and pedigree analysis, YrHA should be a novel resistance gene to stripe rust. The molecular markers of the new resistance gene in H9014-121-5-5-9 could be useful for marker-assisted selection in breeding programs against stripe rust.     

Characterisation and mapping of adult plant stripe rust resistance in wheat accession Aus27284

Key message

A new adult plant stripe rust resistance gene, Yr80, was identified in a common wheat landrace Aus27284. Linked markers were developed and validated for their utility in marker-assisted selection.

Abstract

Stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), is among the most important constraints to global wheat production. The identification and characterisation of new sources of host plant resistance enrich the gene pool and underpin deployment of resistance gene pyramids in new cultivars. Aus27284 exhibited resistance at the adult plant stage against predominant Pst pathotypes and was crossed with a susceptible genotype Avocet S. A recombinant inbred line (RIL) population comprising 121 lines was developed and tested in the field at three locations in 2016 and two in 2017 crop seasons. Monogenic segregation for adult plant stripe rust response was observed among the Aus27284/Avocet S RIL population and the underlying locus was temporarily designated YrAW11. Bulked-segregant analysis using the Infinium iSelect 90K SNP wheat array placed YrAW11 in chromosome 3B. Kompetitive allele specific PCR (KASP) primers were designed for the linked SNPs and YrAW11 was flanked by KASP_65624 and KASP_53292 (3 cM) proximally and KASP_53113 (4.9 cM) distally. A partial linkage map of the genomic region carrying YrAW11 comprised nine KASP and two SSR markers. The physical position of KASP markers in the pseudomolecule of chromosome 3B placed YrAW11 in the long arm and the location of markers gwm108 and gwm376 in the deletion bin 3BL2-0.22 supported this conclusion. As no other stripe rust resistance locus has been reported in chromosome 3BL, YrAW11 was formally designated Yr80. Marker KASP_ 53113 was polymorphic among 94% of 81 Australian wheat cultivars used for validation.

     

Genetic analysis of leaf rust resistance genes and associated markers in the durable resistant wheat cultivar Sinvalocho MA

In the cross of the durable leaf rust resistant wheat Sinvalocho MA and the susceptible line Gama6, four specific genes were identified: the seedling resistance gene Lr3, the adult plant resistance (APR) genes LrSV1 and LrSV2 coming from Sinvalocho MA, and the seedling resistance gene LrG6 coming from Gama6. Lr3 was previously mapped on 6BL in the same cross. LrSV1 was mapped on chromosome 2DS where resistance genes Lr22a and Lr22b have been reported. Results from rust reaction have shown that LrSV1 from Sinvalocho is not the same allele as Lr22b and an allelism test with Lr22a showed that they could be alleles or closely linked genes. LrSV1 was mapped in an 8.5-cM interval delimited by markers gwm296 distal and gwm261 proximal. Adult gene LrSV2 was mapped on chromosome 3BS, cosegregating with gwm533 in a 7.2-cM interval encompassed by markers gwm389 and gwm493, where other disease resistance genes are located, such as seedling gene Lr27 for leaf rust, Sr2 for stem rust, QTL Qfhs.ndsu-3BS for resistance to Fusarium gramineum and wheat powdery mildew resistance. The gene LrG6 was mapped on chromosome 2BL, with the closest marker gwm382 at 0.6 cM. Lines carrying LrSV1, LrSV2 and LrG6 tested under field natural infection conditions, showed low disease infection type and severity, suggesting that this kind of resistance can be explained by additive effects of APR and seedling resistance genes. The identification of new sources of resistance from South American land races and old varieties, supported by modern DNA technology, contributes to sustainability of agriculture through plant breeding.     

Identification and mapping of resistance to stem rust in the European winter wheat cultivars Spark and Rialto

A mapping population of 126 doubled haploid (DH) lines derived from a cross between the English winter wheat cultivars Spark and Rialto was evaluated for response to Puccinia graminis f. sp. tritici in the greenhouse and in artificially inoculated field plots at two locations over 3 years (2011, 2012 and 2013). Genetic analysis indicated the involvement of two seedling genes (Sr5 and Sr31, contributed by Rialto) and three adult plant resistance genes. QTL analyses of field data showed the involvement of three consistent effects QTL on chromosome arms 1BS (contributed by Rialto), and 3BS and chromosome 5A (contributed by Spark) in the observed resistance to stem rust. These QTLs explained average phenotypic variation of 78.5, 9.0 and 5.9 %, respectively. With the presence of virulence for Sr5 and absence of Sr31 virulence in the field, the QTL detected on 1BS (QSr.sun-1BS) was attributed to the major seedling resistance gene Sr31. The QTL located on chromosome arm 3BS (QSr.sun-3BS) was closely associated with SSR marker gwm1034, and the QTL detected on 5A (QSr.sun-5A) was closely linked with SSR marker gwm443. DH lines carrying the combination of QSr.sun-3BS and QSr.sun-5A exhibited lower stem rust responses indicating the additive effects of the two APR genes in reducing disease severity. The markers identified in this study can be useful in pyramiding these QTLs with other major or minor genes and marker assisted selection for stem rust resistance in wheat.     

Resistance to recombinant stem rust race TPPKC in hard red spring wheat

The wheat (Triticum aestivum L.) stem rust (Puccinia graminis Pers.:Pers. f.sp. tritici Eriks. and Henn.) resistance gene SrWld1 conditions resistance to all North American stem rust races and is an important gene in hard red spring (HRS) wheat cultivars. A sexually recombined race having virulence to SrWld1 was isolated in the 1980s. Our objective was to determine the genetics of resistance to the race. The recombinant race was tested with the set of stem rust differentials and with a set of 36 HRS and 6 durum cultivars. Chromosomal location studies in cultivars Len, Coteau, and Stoa were completed using aneuploid analysis, molecular markers, and allelism tests. Stem rust differential tests coded the race as TPPKC, indicating it differed from TPMKC by having added virulence on Sr30 as well as SrWld1. Genes effective against TPPKC were Sr6, Sr9a, Sr9b, Sr13, Sr24, Sr31, and Sr38. Genetic studies of resistance to TPPKC indicated that Len, Coteau, and Stoa likely carried Sr9b, that Coteau and Stoa carried Sr6, and Stoa carried Sr24. Tests of HRS and durum cultivars indicated that five HRS and one durum cultivar were susceptible to TPPKC. Susceptible HRS cultivars were postulated to have SrWld1 as their major stem rust resistance gene. Divide, the susceptible durum cultivar, was postulated to lack Sr13. We concluded that although TPPKC does not constitute a threat similar to TTKSK and its variants, some cultivars would be lost from production if TPPKC became established in the field.     

Marker Assisted Transfer of Two Powdery Mildew Resistance Genes PmTb7A.1 and PmTb7A.2 from Triticum boeoticum (Boiss.) to Triticum aestivum (L.)

Powdery mildew (PM), caused by Blumeria graminis f. sp. tritici, is one of the important wheat diseases, worldwide. Two PM resistance genes, designated as PmTb7A.1 and PmTb7A.2, were identified in T. boeoticum acc. pau5088 and mapped on chromosome 7AL approximately 48cM apart. Two resistance gene analogue (RGA)-STS markers Ta7AL-4556232 and 7AL-4426363 were identified to be linked to the PmTb7A.1 and PmTb7A.2, at a distance of 0.6cM and 6.0cM, respectively. In the present study, following marker assisted selection (MAS), the two genes were transferred to T. aestivum using T. durum as bridging species. As many as 12,317 florets of F₁ of the cross T. durum /T. boeoticum were pollinated with T. aestivum lines PBW343-IL and PBW621 to produce 61 and 65 seeds, respectively, of three-way F₁. The resulting F₁s of the cross T. durum/T. boeoticum//T. aestivum were screened with marker flanking both the PM resistance genes PmTb7A.1 and PmTb7A.2 (foreground selection) and the selected plants were backcrossed to generate BC₁F₁. Marker assisted selection was carried both in BC₁F₁ and the BC₂F₁ generations. Introgression of alien chromatin in BC₂F₁ plants varied from 15.4 - 62.9 percent. Out of more than 110 BC₂F₁ plants showing introgression for markers linked to the two PM resistance genes, 40 agronomically desirable plants were selected for background selection for the carrier chromosome to identify the plants with minimum of the alien introgression. Cytological analysis showed that most plants have chromosome number ranging from 40-42. The BC₂F₂ plants homozygous for the two genes have been identified. These will be crossed to generate lines combining both the PM resistance genes but with minimal of the alien introgression. The PM resistance gene PmTb7A.1 maps in a region very close to Sr22, a stem rust resistance gene effective against the race Ug99. Analysis of selected plants with markers linked to Sr22 showed introgression of Sr22 from T. boeoticum in several BC₂F₁ plants. Thus, in addition to PM resistance, these progeny might also carry resistance to stem rust race Ug99.     

High-resolution mapping and mutation analysis separate the rust resistance genes Sr31, Lr26 and Yr9 on the short arm of rye chromosome 1

The stem, leaf and stripe rust resistance genes Sr31, Lr26 and Yr9, located on the short arm of rye chromosome 1, have been widely used in wheat by means of wheat-rye translocation chromosomes. Previous studies have suggested that these resistance specificities are encoded by either closely-linked genes, or by a single gene capable of recognizing all three rust species. To investigate these issues, two 1BL·1RS wheat lines, one with and one without Sr31, Lr26 and Yr9, were used as parents for a high-resolution F2 mapping family. Thirty-six recombinants were identified between two PCR markers 2.3 cM apart that flanked the resistance locus. In one recombinant, Lr26 was separated from Sr31 and Yr9. Mutation studies recovered mutants that separated all three rust resistance genes. Thus, together, the recombination and mutation studies suggest that Sr31, Lr26 and Yr9 are separate closely-linked genes. An additional 16 DNA markers were mapped in this region. Multiple RFLP markers, identified using part of the barley Mla powdery mildew resistance gene as probe, co-segregated with Sr31 and Yr9. One deletion mutant that had lost Sr31, Lr26 and Yr9 retained all Mla markers, suggesting that the family of genes on 1RS identified by the Mla probe does not contain the Sr31, Lr26 or Yr9 genes. The genetic stocks and DNA markers generated from this study should facilitate the future cloning of Sr31, Lr26 and Yr9.     

Genetic mapping of adult plant leaf rust resistance genes Lr48 and Lr49 in common wheat

Hypersensitive adult plant resistance genes Lr48 and Lr49 were named based on their genetic independence of the known adult plant resistance genes. This study was planned to determine genomic locations of these genes. Recombinant inbred line populations derived from crosses involving CSP44 and VL404, sources of Lr48 and Lr49, respectively, and the susceptible parent WL711, were used to determine the genomic locations of these genes. Bulked segregant analyses were performed using multiplex-ready PCR technology. Lr48 in genotype CSP44 was mapped on chromosome arm 2BS flanked by marker loci Xgwm429b (6.1 cM) and Xbarc7 (7.3 cM) distally and proximally, respectively. Leaf rust resistance gene Lr13, carried by the alternate parent WL711, was proximal to Lr48 and was flanked by Xksm58 (5.1 cM) and Xstm773-2 (8.7 cM). Lr49 was flanked by Xbarc163 (8.1 cM) and Xwmc349 (10.1 cM) on chromosome arm 4BL. The likely presence of the durable leaf rust resistance gene Lr34 in both CSP44 and VL404 was confirmed using the tightly linked marker csLV34. Near-isogenic lines for Lr48 and Lr49 were developed in cultivar Lal Bahadur. Genotypes combining Lr13 and/or Lr34 with Lr48 or Lr49 were identified as potential donor sources for cultivar development programs.     

Development of co-dominant KASP markers co-segregating with Ug99 effective stem rust resistance gene <Emphasis Type="Italic">Sr26</Emphasis> in wheat

Stem rust of wheat, caused by Puccinia graminis f. sp. tritici (Pgt), is a threat to global food security due to its ability to cause total crop failures. The Pgt race TTKSK (Ug99) and its derivatives detected in East Africa carry virulence for many resistance genes present in modern cultivars. However, stem rust resistance gene Sr26 remains effective to all races of Pgt worldwide. Sr26 is carried on the Agropyron elongatum (syn. Thinopyrum ponticum) segment 6Ae#1L translocated to chromosome 6AL of wheat. In this study, a recombinant inbred line (RIL) population derived from a cross between the landrace Aus27969 and Avocet S, which carries Sr26, was used to develop co-dominant kompetitive allele-specific polymerase chain reaction (KASP) markers that co-segregate with Sr26. Four KASP markers (sunKASP_216, sunKASP_218, sunKASP_224 and sunKASP_225) were also shown to co-segregate with Sr26 in four additional RIL populations. When tested on Australian cultivars and breeding lines, these markers amplified alleles alternate to that linked with Sr26 in all cultivars known to lack this gene and Sr26-linked alleles in cultivars and genotypes known to carry Sr26. Genotypes WA-1 and WA-1/3*Yitpi carrying the shortest Sr26 translocation segment were positive only for markers sunKASP_224 and sunKASP_225. Our results suggest the four KASP markers are located on the original translocation and sunKASP_224 and sunKASP_225 are located on the shortened version. Therefore, sunKASP_224 and sunKASP_225 can be used for marker-assisted pyramiding of Sr26 with other stem rust resistance genes to achieve durable resistance in wheat.     

Molecular mapping of a new temperature-sensitive gene <Emphasis Type="Italic">LrZH22</Emphasis> for leaf rust resistance in Chinese wheat cultivar Zhoumai 22

Leaf rust, caused by Puccinia triticina, is one of the most widespread diseases in common wheat globally. The Chinese wheat cultivar Zhoumai 22 is highly resistant to leaf rust at the seedling and adult stages. Seedlings of Zhoumai 22 and 36 lines with known leaf rust resistance genes were inoculated with 13 P. triticina races for gene postulation. The leaf rust response of Zhoumai 22 was different from those of the single gene lines. With the objective of identifying and mapping, the new gene(s) for resistance to leaf rust, F₁, F₂ plants and F_2:3 lines from the cross Zhoumai 22/Chinese Spring were inoculated with Chinese P. triticina race FHDQ at the seedling stage. A single dominant gene, tentatively designated LrZH22, conferred resistance. To identify other possible genes in Zhoumai 22, ten P. triticina races avirulent on Zhoumai 22 were used to inoculate 24 F_2:3 lines. The same gene conferred resistance to all ten avirulent races. A total of 1300 simple sequence repeat (SSR) markers and 36 EST markers on 2BS were used to test the parents, and resistant and susceptible bulks. Resistance gene LrZH22 was mapped in the chromosome bin 2BS1-0.53-0.75 and closely linked to six SSR markers (barc183, barc55, gwm148, gwm410, gwm374 and wmc474) and two EST markers (BF202681 and BE499478) on chromosome arm 2BS. The two closest flanking SSR loci were Xbarc55 and Xgwm374 with genetic distances of 2.4 and 4.8 cM from LrZH22, respectively. Six designated genes (Lr13, Lr16, Lr23, Lr35, Lr48 and Lr73) are located on chromosome arm 2BS. In seedling tests, LrZH22 was temperature sensitive, conferring resistance at high temperatures. The reaction pattern of Zhoumai 22 was different from that of RL 4031 (Lr13), RL 6005 (Lr16) and RL 6012 (Lr23), Lr35 and Lr48 are adult-plant resistance genes, and Lr73 is not sensitive to the temperature. Therefore, LrZH22 is likely to be a new leaf rust resistance gene or allele.     

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.     

Adult plant stripe rust resistance gene <Emphasis Type="Italic">Yr71</Emphasis> maps close to <Emphasis Type="Italic">Lr24</Emphasis> in chromosome 3D of common wheat

Australian cultivar Sunco carries three adult plant stripe rust resistance genes. One of these genes corresponded to Yr18 in chromosome 7DS; the second, YrCK, was mapped on chromosome 2D. Here, we describe the characterization of the third adult plant resistance (APR) gene from Sunco. Sunco/2*Avocet S-derived lines SA65 (resistant) and SA67 (susceptible) were crossed and a recombinant inbred line F₆ population was generated. Monogenic segregation among SA65/SA67-derived RIL population was demonstrated and the resistance locus was designated YrSA3. Selective genotyping using an iSelect 90 K Infinium SNP array and SSR markers located YrSA3 on chromosome 3D. Development of KASP markers for SNP loci showing association with YrSA3 allowed construction of a genetic map harboring the resistance gene. Ten KASP markers (KASP_8306, KASP_9142, KASP_10438, KASP_16434, KASP_17207, KASP_20836, KASP_23518, KASP_23615, KASP_57983 and KASP_63653), one SSR marker (gwm114b) and Lr24/Sr24 were mapped 1.8 cM distal to YrSA3. Comparison of marker data indicated that the previously named seedling stripe rust resistance gene Yr45 was located proximal to YrSA3, and therefore the latter was formally designated Yr71. Two recombinants carrying Lr24/Sr24 and Yr71 in combination were identified for use as donor sources in wheat breeding programs. The robustness of gwm114b, KASP_16434, KASP_17207 and KASP_20836 for marker-assisted selection of these genes was demonstrated through tests on 74 Australian wheat cultivars.     

Development of wheat–Aegilops speltoides recombinants and simple PCR-based markers for Sr32 and a new stem rust resistance gene on the 2S#1 chromosome

Key message

Wheat– Aegilops speltoides recombinants carrying stem rust resistance genes Sr32 and SrAes1t effective against Ug99 and PCR markers for marker-assisted selection.

Abstract

Wild relatives of wheat are important resources for new rust resistance genes but underutilized because the valuable resistances are often linked to negative traits that prevent deployment of these genes in commercial wheats. Here, we report ph1b-induced recombinants with reduced alien chromatin derived from E.R. Sears’ wheat–Aegilops speltoides 2D-2S#1 translocation line C82.2, which carries the widely effective stem rust resistance gene Sr32. Infection type assessments of the recombinants showed that the original translocation in fact carries two stem rust resistance genes, Sr32 on the short arm and a previously undescribed gene SrAes1t on the long arm of chromosome 2S#1. Recombinants with substantially shortened alien chromatin were produced for both genes, which confer resistance to stem rust races in the TTKSK (Ug99) lineage and representative races of all Australian stem rust lineages. Selected recombinants were back crossed into adapted Australian cultivars and PCR markers were developed to facilitate the incorporation of these genes into future wheat varieties. Our recombinants and those from several other labs now show that Sr32, Sr39, and SrAes7t on the short arm and Sr47 and SrAes1t on the long arm of 2S#1 form two linkage groups and at present no rust races are described that can distinguish these resistance specificities.