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Heat stress at later growth stage is one of the major limitations in achieving potential yield in barley. Incorporation of heat tolerance in the variety development process is an essential task that breeders would like to achieve by exploring new sources of genetic variability and their utilization. A better understanding of genetic variation in existing genotypes under heat stress is required to produce high yielding varieties with improved heat tolerance. An association mapping panel 2017 (AM2017), comprising of 316 genotypes, was evaluated under timely and late sown (heat stress) conditions for two consecutive crop seasons at Hisar in India during 2017–2019. Eight agro-morphological traits, mainly contributing to yield, were considered agro-morphological diversity study. Genetic diversity and population structure were explored by using the 50 K iSelect Illumina Barley SNP array. A set of 36,793 SNP markers, covering a genetic distance of 991.82 cM with an average marker density of 37.09 SNPs/cM, was obtained after quality filtration. The gene diversity (GD) and Polymorphic Information Content (PIC) at genome level was 0.362 and 0.289, respectively. In AM2017, two subpopulations were observed mainly due to the row types. Principal component analysis of agro-morphological traits revealed that days to heading and maturity along with spike length, spikelet number per spike and grain yield per plot were the most important traits in timely and late sown conditions. The information on agro-morphological genetic diversity under heat stress conditions will be useful in identifying heat tolerant genotypes for use in barley breeding programs.

We performed homoeologous recombination-based partitioning and physical mapping of wheat chromosome 3B and Th. elongatum chromosome 3E, providing a unique physical framework of this homoeologous pair for genome studies. The wheat (Triticum aestivum, 2n = 6x = 42, AABBDD) and Thinopyrum elongatum (2n = 2x = 14, EE) genomes can be differentiated from each other by fluorescent genomic in situ hybridization (FGISH) as well as molecular markers. This has facilitated homoeologous recombination-based partitioning and engineering of their genomes for physical mapping and alien introgression. Here, we constructed a special wheat genotype, which was double monosomic for wheat chromosome 3B and Th. elongatum chromosome 3E and homozygous for the ph1b mutant, to induce 3B-3E homoeologous recombination. Totally, 81 3B-3E recombinants were recovered and detected in the primary, secondary, and tertiary homoeologous recombination cycles by FGISH. Comparing to the primary recombination, the secondary and tertiary recombination shifted toward the proximal regions due to the increase in homology between the pairing partners. The 3B-3E recombinants were genotyped by high-throughput wheat 90-K single nucleotide polymorphism (SNP) arrays and their recombination breakpoints physically mapped based on the FGISH patterns and SNP results. The 3B-3E recombination physically partitioned chromosome 3B into 38 bins, and 429 SNPs were assigned to the distinct bins. Integrative analysis of FGISH and SNP results led to the construction of a composite bin map for chromosome 3B. Additionally, we developed 22 SNP-derived semi-thermal asymmetric reverse PCR markers specific for chromosome 3E and constructed a comparative map of homoeologous chromosomes 3E, 3B, 3A, and 3D. In summary, this work provides a unique physical framework for further studies of the 3B-3E homoeologous pair and diversifies the wheat genome for wheat improvement.

Wheat landrace accessions were chosen from areas of the world with historical European wheat stem sawfly (Cephus pygmaeus L.) selection pressure to develop six recombinant inbred line (RIL) populations. Molecular maps were constructed, and resistance due to antibiosis and antixenosis was assessed at sites in Montana naturally infested by Cephus cinctus Norton, the wheat stem sawfly (WSS). Novel QTLs were identified along with QTL previously identified in elite germplasm. A newly identified QTL on chromosome 1B provided a new source for pith-filled solid stems. An allele for resistance on chromosome 4A unrelated to solid stems was identified in four of the six RIL populations. A landrace from Turkey, PI 166471, contained alleles at three QTLs causing high levels of larval mortality. None of the QTLs were related to stem solidness, but their combined effect provided resistance similar to that observed in a solid-stemmed check cultivar. These results show the utility of genetic populations derived from geographically targeted landrace accessions to identify new alleles for insect resistance. New PCR-based molecular markers were developed for introgression of novel alleles for WSS resistance into elite lines. Comparison of results with previous analysis of elite cultivars addresses changes in allele frequencies during the wheat breeding process.

Barley stripe rust, Puccinia striiformis f. sp. hordei, (PSH) occurs worldwide and is one of the major barley diseases of South Asia and North Africa. The aim of this study was to identify and map rust resistance loci at the seedling and the adult plant stages in 336 barley genotypes assembled at ICARDA as AM-2014. The seedling resistance to PSH was evaluated in the greenhouse individually for six virulent races originated from India. The adult stage resistance to PSH was evaluated in field located at Durgapura, Rajasthan, India. The AM-2014 panel was genotyped with infinium iSelect 9K barley SNP markers and genome-wide association studies were carried out using mixed linear model accounting for population structure (PCA) and kinship (K) as covariates. Eleven barley genotypes were identified resistant to all six PSH races at the seedling stage while these genotypes were also resistant at adult plant stage in the field. The most effective SNP markers (at least one for each QTL) associated with rust resistance was also identified using stepwise regression. A total of 17 and 25 resistance QTL to six races of PSH were identified in two- and six-rowed barley at the seedling stage, respectively. While, six and seven resistance QTL to PSH were detected at the adult stage in two- and six-rowed barley, respectively. The novel PSH resistant QTL detected at the seedling and adult stages are important for barley breeding programs globally.

We physically dissected and mapped wheat chromosome 2B and its homoeologues in Aegilops speltoides and Thinopyrum elongatum based on meiotic homoeologous recombination, providing a unique physical framework for genome studies. Common wheat has a large and complex genome with narrow genetic diversity and various degrees of recombination between the A, B, and D subgenomes. This has limited the homologous recombination-based genome studies in wheat. Here, we exploited meiotic homoeologous recombination for molecular mapping of wheat chromosome 2B and its homoeologue 2S from Aegilops speltoides and 2E from Thinopyrum elongatum. The 2B-2S and 2B-2E recombination was induced by the ph1b mutant, and recovered using molecular markers and fluorescent genomic in situ hybridization (FGISH). A total of 112 2B-2S and 87 2B-2E recombinants involving different chromosome regions were developed and physically delineated by FGISH. The 2B-2S and 2B-2E recombination hotspots mapped to the subterminal regions on both arms. Recombination hotspots with the highest recombination rates mapped to the short arms. Eighty-three 2B-2S and 67 2B-2E recombinants were genotyped using the wheat 90 K SNP arrays. Based on the genotyping results and FGISH patterns of the recombinants, chromosomes 2B, 2S, and 2E were partitioned into 93, 66, and 46 bins, respectively. In total, 1037 SNPs physically mapped onto distinct bins of these three homoeologous chromosomes. A homoeologous recombination-based bin map was constructed for chromosome 2B, providing a unique physical framework for genome studies in wheat and its relatives. Meiotic homoeologous recombination also facilitates gene introgression to diversify the wheat genome for germplasm development. Therefore, homoeologous recombination-based studies enhance understanding of the wheat genome and its homoeologous counterparts from wild grasses, and expand the genetic variability of the wheat genome.

The major QTL for FHB resistance from hexaploid wheat line PI 277012 was successfully introgressed into durum wheat and minor FHB resistance QTL were detected in local durum wheat cultivars. A combination of these QTL will enhance FHB resistance of durum wheat. Fusarium head blight (FHB), caused by Fusarium graminearum, is a devastating disease of durum wheat. To combat the disease, great efforts have been devoted to introgress FHB resistance from its related tetraploid and hexaploid wheat species into adapted durum cultivars. However, most of the quantitative trait loci (QTL) for FHB resistance existing in the introgression lines are not well characterized or validated. In this study, we aimed to identify and map FHB resistance QTL in a population consisting of 205 recombinant inbred lines from the cross between Joppa (a durum wheat cultivar) and 10Ae564 (a durum wheat introgression line with FHB resistance derived from the hexaploid wheat line PI 277012). One QTL (Qfhb.ndwp-2A) from Joppa and two QTL (Qfhb.ndwp-5A and Qfhb.ndwp-7A) from 10Ae564 were identified through phenotyping of the mapping population for FHB severity and DON content in greenhouse and field and genotyping with 90K wheat Infinium iSelect SNP arrays. Qfhb.ndwp-2A explained 14, 15, and 9% of the phenotypic variation, respectively, for FHB severity in two greenhouse experiments and for mean DON content across the two greenhouse environments. Qfhb.ndwp-5A explained 19, 10, and 7% of phenotypic variation, respectively, for FHB severity in one greenhouse experiment, mean FHB severity across two field experiments, and mean DON content across the two greenhouse experiments. Qfhb.ndwp-7A was only detected for FHB severity in the two greenhouse experiments, explaining 9 and 11% of the phenotypic variation, respectively. This study confirms the existence of minor QTL in North Dakota durum cultivars and the successful transfer of the major QTL from PI 277012 into durum wheat.

Wheat cultivar Madsen has a new gene on the short arm of chromosome 1A and two QTL for all-stage resistance and three QTL for high-temperature adult-plant resistance that in combination confer high-level, durable resistance to stripe rust. Wheat cultivar Madsen has maintained a high-level resistance to stripe rust over 30 years. To map quantitative trait loci (QTL) underlying the high-level, durable resistance, 156 recombinant inbred lines (RILs) developed from cross Avocet S × Madsen were phenotyped with selected races of Puccinia striiformis f. sp. tritici in the greenhouse seedling tests, and in naturally infected fields during 2015–2017. The RILs were genotyped by SSR and SNP markers from genotyping by sequencing and the 90 K wheat SNP chip. Three QTL for all-stage resistance were mapped on chromosomes 1AS, 1BS and 2AS, and two QTL for high-temperature adult-plant (HTAP) resistance were mapped on 3BS and 6BS. The most effective QTL on 2AS, explaining 8.97–23.10% of the phenotypic variation in seedling tests and 8.60–71.23% in field tests, contained Yr17 for all-stage resistance and an additional gene for HTAP resistance. The 6BS QTL, detected in all field tests, was identified as Yr78. The 1AS QTL, conferring all-stage resistance, was identified as a new gene, which explained 20.45 and 30.23% of variation in resistance to races PSTv-37 and PSTv-40, respectively, and contributed significantly to field resistance at Pullman in 2015-2017, but was not detected at Mount Vernon. The interactions among QTL were mostly additive, and RILs with all five QTL had the highest level of resistance in the field, similar to Madsen. Genotyping 148 US Pacific Northwest wheat cultivars with markers for the 1AS, 2AS and 6BS QTL validated the genes and markers, and indicated their usefulness for marker-assisted selection.

Genetic diversity of 139 accessions of diploid Triticum species including Triticum urartu, Triticum boeoticum and Triticum monococcum was studied using 11 SSR (simple sequence repeats) markers. A total of 111 alleles with an average of 10 alleles per locus were detected. The polymorphism information content (PIC) of each SSR marker ranged from 0.30 to 0.90 with an average value of 0.62. Among the three Triticum species T. urartu had the highest number of total alleles (Na = 81), private alleles (Npa = 15) and showed higher genetic diversity (Hex = 0.58; PIC = 0.54). The genotypes from Turkey exhibited the highest genetic diversity (PIC = 0.6), while the least diversity was observed among 4 Georgian accessions (PIC = 0.11). Cluster analysis was able to distinguish 139 wheat accessions at the species level. The highest genetic similarity (GS) was noted between T. boeticum and T. monococcum (GS = 0.84), and the lowest between T. urartu and T. monococcum (GS = 0.46). The grouping pattern of the PCoA analysis corresponded with cluster analysis. No significant differences were found in clustering of T. urartu and T. monococcum accessions with respect to their geographic regions, while within T. boeoticum species, accessions from Iran were somewhat associated with their geographical origin and clustered as a close and separate group. The results from our study demonstrated that SSR markers were good enough for further genetic diversity analysis in einkorn wheat species.

This work pinpointed the goatgrass chromosomal segment in the wheat B genome using modern cytogenetic and genomic technologies, and provided novel insights into the origin of the wheat B genome. Wheat is a typical allopolyploid with three homoeologous subgenomes (A, B, and D). The donors of the subgenomes A and D had been identified, but not for the subgenome B. The goatgrass Aegilops speltoides (genome SS) has been controversially considered a possible candidate for the donor of the wheat B genome. However, the relationship of the Ae. speltoides S genome with the wheat B genome remains largely obscure. The present study assessed the homology of the B and S genomes using an integrative cytogenetic and genomic approach, and revealed the contribution of Ae. speltoides to the origin of the wheat B genome. We discovered noticeable homology between wheat chromosome 1B and Ae. speltoides chromosome 1S, but not between other chromosomes in the B and S genomes. An Ae. speltoides-originated segment spanning a genomic region of approximately 10.46 Mb was detected on the long arm of wheat chromosome 1B (1BL). The Ae. speltoides-originated segment on 1BL was found to co-evolve with the rest of the B genome. Evidently, Ae. speltoides had been involved in the origin of the wheat B genome, but should not be considered an exclusive donor of this genome. The wheat B genome might have a polyphyletic origin with multiple ancestors involved, including Ae. speltoides. These novel findings will facilitate genome studies in wheat and other polyploids.

Plant breeders are interested in using diverse genotypes in hybridization that can segregate for traits of importance with possibility of selection and genetic gain. Information on molecular and agro-morphological diversity helps the breeders reduce the effort for parental selection and helps the advancement of generations. A phenotypic and molecular diversity study, using 24 traits (agronomic and disease) and 6519 SNPs in a diverse collection of 336 spring barley genotypes, was carried out at Marchouch and Jemma Shiam research stations in Morocco. Based on structure and multivariate analyses, strong differentiation between the two- and six-row types were observed. The linkage disequilibrium (LD) decay of the current collection (for the combined population) was up to 3.58 cM (r 2 = 0.15) while LD decay were estimated 3.91 and 2.36 cM for two- and six-row barley, respectively. PCA of agro-morphological traits revealed grain per spike, net form of net blotch (NFNB), spot form of net blotch (SFNB), and 1000 kernel weight were the most discriminatory traits in the current collection. Association mapping in the two independent populations will be ideal for identification of markers, and QTL related to traits. The generated information on relatedness between individuals will help identify diverse genotypes for breeding programs.

A CIho 5791 × Tifang recombinant inbred mapping population was developed and used to identify major dominant resistance genes on barley chromosomes 6H and 3H in CI5791 and on 3H in Tifang. The barley line CIho 5791 confers high levels of resistance to Pyrenophora teres f. teres, causal agent of net form net blotch (NFNB), with few documented isolates overcoming this resistance. Tifang barley also harbors resistance to P. teres f. teres which was previously shown to localize to barley chromosome 3H. A CIho 5791 × Tifang F6 recombinant inbred line (RIL) population was developed using single seed descent. The Illumina iSelect SNP platform was used to identify 2562 single nucleotide polymorphism (SNP) markers across the barley genome, resulting in seven linkage maps, one for each barley chromosome. The CIho 5791 × Tifang RIL population was evaluated for NFNB resistance using nine P. teres f. teres isolates collected globally. Tifang was resistant to four of the isolates tested whereas CIho 5791 was highly resistant to all nine isolates. QTL analysis indicated that the CIho 5791 resistance mapped to chromosome 6H whereas the Tifang resistance mapped to chromosome 3H. Additionally, CIho 5791 also harbored resistance to two Japanese isolates that mapped to a 3H region similar to that of Tifang. SNP markers and RILs harboring both 3H and 6H resistance will be useful in resistance breeding against NFNB.

Based on the estimates of accuracy, genomic selection would be useful for selecting for improved trait values and trait stability for agronomic and quality traits in wheat. Trait values and trait stability estimated by two methods were generally independent indicating a breeder could select for both simultaneously. Genomic selection (GS) is a new marker-assisted selection tool for breeders to achieve higher genetic gain faster and cheaper. Breeders face challenges posed by genotype by environment interaction (GEI) pattern and selecting for trait stability. Obtaining trait stability is costly, as it requires data from multiple environments. There are few studies that evaluate the efficacy of GS for predicting trait stability. A soft winter wheat population of 273 lines was genotyped with 90 K single nucleotide polymorphism markers and phenotyped for four agronomic and seven quality traits. Additive main effect and multiplicative interaction (AMMI) model and Eberhart and Russell regression (ERR) were used to estimate trait stability. Significant GEI variation was observed and stable lines were identified for all traits in this study. The accuracy of GS ranged from 0.33 to 0.67 for most traits and trait stability. Accuracy of trait stability was greater than trait itself for yield (0.44 using AMMI versus 0.33) and heading date (0.65 using ERR versus 0.56). The opposite trend was observed for the other traits. GS did not predict the stability of the quality traits except for flour protein, lactic acid and softness equivalent. Significant GS accuracy for some trait stability indicated that stability was under genetic control for these traits. The magnitude of GS accuracies for all the traits and most of the trait stability index suggests the possibility of rapid selection for these trait and trait stability in wheat breeding.

Spot blotch (SB) in barley is caused by the fungal pathogen Cochliobolus sativus and considered one of the major constraints to successful barley production. Resistance to C. sativus was evaluated, using a barley collection of 336 genotypes (AM-2014), at the seedling and adult stages. Seedling resistance was evaluated by using a mixture of 19 virulent isolates in Morocco. Virulent isolates prevalent in Uttar Pradesh were used for phenotyping resistance at the adult stage in India. The AM-2014 panel was genotyped with 9-K single-nucleotide polymorphism (SNP) markers using iSelect Illumina Infinium. Genome wide association studies (GWAS) were carried out using SNP markers, infection responses, disease severity, and area under the disease progress curve (AUDPC). The mixed linear model was employed in TASSEL using principal component analysis (PCA) and Kinship matrix (K) as covariates. Higher SB severity, 82.3 ± 13.5 (mean ± SD), was recorded at the Banaras Hindu University (BHU) compared to 47.6 ± 15.0 at the Narendra Dev University of Agriculture and Technology (NDUAT). Nine QTL, Rcs-qtl-1H-126.9, Rcs-qtl-2H-148.16, Rcs-qtl-3H-25.27, Rcs-qtl-5H-80.35, Rcs-qtl-6H-58.24, Rcs-qtl-7H-29.62, Rcs-qtl-7H-29.72, Rcs-qtl-7H-32.81, and Rcs-qtl-7H-34.74, were detected for SB resistance at the seedling stage. For SB severity at the adult stage, a QTL, Rcs-qtl-7H-32.81, was detected at BHU while seven QTL, Rcs-qtl-2H-91.09, Rcs-qtl-3H-145.64, Rcs-qtl-4H-14.43, Rcs-qtl-6H-6.49, Rcs-qtl-7H-114.43, Rcs-qtl-7H-151.66, and Rcs-qtl-7H-150.36, were found for SB severity at NDUAT. Three QTL, Rcs-qtl-4H-18.61, Rcs-qtl-4H-67.91, and Rcs-qtl-5H-110.25, were significant for AUDPC of SB at BHU. The QTLs reported in this study are important to advance marker-assisted selection and gene pyramiding of SB resistance in South Asia and North Africa in future.

Net form of net blotch (NFNB) of barley (Hordeum vulgare L.), caused by Pyrenophora teres f. teres (Ptt) Drechsler (anamorph: Drechslera teres [Sacc.] Shoem.), is considered one of the major constraints of successful barley production in major barley growing regions of the world. Resistance to NFNB was evaluated in a barley collection of 336 genotypes (AM-2014), at seedling stage using isolates LGDPtt.19 and TD10 in the USA, and adult stage in seven hotspot environments in Morocco. The AM-2014 panel was genotyped with 9K SNP markers and genome-wide association studies (GWAS) were carried out using mixed linear model (MLM: Q + K) accounting for population structure (Q) and kinship (K) as covariates. Significant (P

Durum wheat is capable of accumulating cadmium, a toxic heavy metal, in the grain at levels that have been deemed unsafe for human consumption. Previous studies have identified genetic variation as well as markers associated with Cd accumulation in durum wheat, which can be exploited to develop low Cd cultivars. Because the phenotyping for Cd content is very expensive, KASP markers were developed from molecular markers associated with grain Cd and tested for their usefulness for marker-assisted breeding. A total of 1278 unique genotypes from preliminary and advanced yield trials grown at multiple locations for 2 years were evaluated for grain Cd as well as screened for markers associated with Cd uptake. One marker on chromosome 5B was polymorphic in all crosses between high and low Cd parents and had r2 values ranging from 0.38 to 0.85. Two other markers on the same chromosome predicted similar levels of variation in many trials; however, they were not polymorphic in all populations. The KASP markers accurately predicted up to 97% of the lines for Cd phenotype in different trials. This study identified two markers, Cad-5B and Ex_c1343_2570756, with an average prediction accuracy of 84–88%. These markers could be useful for marker-assisted selection for low grain Cd in durum wheat.

The impact of the D genome and QTL in the A and B genomes on agronomic performance of hexaploid wheat and tetraploid durum was determined using novel recombinant inbred line populations derived from interploid crosses. Genetic differences between common hexaploid (6X) bread wheat (Triticum aestivum, 2n = 6x = 42, genome, AABBDD) and tetraploid (4X) durum wheat (T. turgidum subsp. durum, 2n = 4x = 28, genome, AABB) may exist due to effects of the D genome and allelic differences at loci in the A and B genomes. Previous work allowed identification of a 6X by 4X cross combination that resulted in a large number of fertile recombinant progeny at both ploidy levels. In this study, interspecific recombinant inbred line populations at both 4X and 6X ploidy with 88 and 117 individuals, respectively, were developed from a cross between Choteau spring wheat (6X) and Mountrail durum wheat (4X). The presence of the D genome in the 6X population resulted in increased yield, tiller number, kernel weight, and kernel size, as well as a decrease in stem solidness, test weight and seed per spike. Similar results were found with a second RIL population containing 152 lines from 18 additional 6X by 4X crosses. Several QTL for agronomic and quality traits were identified in both the 4X and 6X populations. Although negatively impacted by the lack of the D genome, kernel weight in Mountrail (4X) was higher than Choteau (6X) due to positive alleles from Mountrail on chromosomes 3B and 7A. These and other favorable alleles may be useful for introgression between ploidy levels.

Continuous improvement in grain yield is one of the major challenges for wheat (Triticum aestivum L.) breeding worldwide. This study characterized quantitative trait loci (QTL) underlying wheat grain yield and its components using a high-density genetic linkage map developed from a recombinant inbred line (RIL) population derived from ‘Ning7840’ × ‘Clark’. The map consisted of 594 single nucleotide polymorphism and 404 simple sequence repeat markers covering a genetic distance of 4225.7 cM. The RIL population was evaluated for grain yield (GY), spike number per m2 (SNPM), kernel number per spike (KNPS), and thousand-kernel weight (TKW) in three Oklahoma locations from 2001 to 2003. A total of 29 additive QTL (eight for GY, two for SNPM, five for KNPS, and 14 for TKW) were mapped on 13 chromosomes. Eight pairs of epistatic QTL were detected for different yield components: four for GY, two for KNPS, and two for TKW. Four additive QTL, including two for GY and two for KNPS, showed additive × environment interactions. QTL that were repeatable in multiple environments were identified for all traits except SNPM. Positive alleles were dispersed between the two parents for all traits, with ‘Clark’ contributing slightly more. Seven pleiotropic loci were co-localized for at least two traits. Interestingly, all co-localized loci overlapped for TKW, and four of them overlapped for GY. Thus, selecting QTL for TKW may simultaneously select for or against yield or other yield components in breeding.

Fusarium head blight (FHB), caused by Fusarium graminearum, threatens durum wheat (Triticum turgidum subsp. durum) production in many durum-growing regions. It is critical to identify useful sources of FHB resistance for durum wheat. A domesticated emmer wheat (T. turgidum subsp. dicoccum) accession, PI 41025, was previously shown to be moderately resistant to FHB. This study aimed to identify quantitative trait loci (QTL) associated with FHB resistance in PI 41025. A population of 200 recombinant inbred lines were developed from a cross between the durum cultivar ‘Ben’ and PI 41025 and evaluated for reactions to F. graminearum in one field nursery and three greenhouse experiments. The disease severity data and a single nucleotide polymorphism marker-based linkage map from this population were used for QTL analysis. The results showed that one QTL on chromosome 2A from Ben and one QTL each on 3A and 5A from PI 41025 were associated with FHB resistance. The 2A and 3A QTL each explaining 8 % of the phenotypic variation were detected only in the greenhouse experiments. The QTL on 5A, which mapped near the domestication gene Q, explained 11 and 35 % of phenotypic variation in greenhouse and field evaluations, respectively. The identification of the 2A QTL from Ben confirmed the presence of FHB resistance in North Dakota durum cultivars, which have been successfully used for developing new cultivars with improved FHB tolerance. This study indicates that combining the QTL from related tetraploid species with native durum QTL will be useful for improving FHB resistance in durum.

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

In plant species, construction of framework linkage maps to facilitate quantitative trait loci mapping and molecular breeding has been confined to experimental mapping populations. However, development and evaluation of these populations is detached from breeding efforts for cultivar development. In this study, we demonstrate that dense and reliable linkage maps can be constructed using extant breeding populations derived from a large number of crosses, thus eliminating the need for extraneous population development. Using 565 segregating F1 progeny from 28 four-way cross breeding populations, a linkage map of the hexaploid wheat genome consisting of 3,785 single nucleotide polymorphism (SNP) loci and 22 simple sequence repeat loci was developed. Map estimation was facilitated by application of mapping algorithms for general pedigrees implemented in the software package CRI-MAP. The developed linkage maps showed high rank-order concordance with a SNP consensus map developed from seven mapping studies. Therefore, the linkage mapping methodology presented here represents a resource efficient approach for plant breeding programs that enables development of dense linkage maps “on the fly” to support molecular breeding efforts.

A renewed interest in breeding barley specifically for food end-uses is being driven by increased consumer interest in healthier foods. We conducted association mapping on physicochemical properties of barley that play a role in food quality and processing including grain hardness, polyphenol oxidase activity, total phenolics, amylose content, and β-glucan. We used 3,069 elite two-row and six-row spring barley breeding lines from eight US breeding programs and 2,041 SNP markers for association mapping. Marker–trait associations were identified using a mixed model that incorporated population structure and kinship. We detected two previously identified QTL for grain hardness on chromosome 2H in the telomeric region of 5H along with two novel regions on 4H and 6H. For amylose content, we detected marker–trait associations on 7H from 0.63 to 30 cM. We detected four regions on chromosomes 1H, 2H, 3H, and 4H associated with polyphenol oxidase activity. The chromosome 2H region co-localized with the two previously mapped polyphenol oxidase genes PPO1 and PPO2, and the regions on chromosomes 1H, 3H, and 4H QTL were novel. For total phenolics, we identified three significant regions on 3H, 4H, and 5H. Two regions on 2H and 7H were associated with β-glucan. Both previously identified and novel QTL are segregating in elite US breeding germplasm. Only three of the 24 SNPs that were associated with traits using either the two-row or six-row mapping panel were identified in both panels. Nine SNPs were detected in the individual two-row or six-row panels that were not detected in the analysis using the complete panel and accounting for population structure. The distribution of favorable alleles at these loci that underpin food quality across the breeding programs suggests several strategies to use markers to improve barley for food uses.

Managing wheat stem rust, caused by Puccinia graminis f. sp. tritici (Pgt), is imperative for the preservation of global food security. The most effective strategy is pyramiding several resistance genes into adapted wheat cultivars. A search for new resistance sources to Pgt race TTKSK resistance identified a spring wheat landrace, accession PI 626573, as a potentially novel source of resistance. A cross was made between LMPG-6, a susceptible spring wheat line, and PI 626573 and used to develop a recombinant inbred population to map the resistance. Bulk segregant analysis (BSA) of LMPG-6/PI 626573 F2 progeny determined resistance was conferred by a single dominant gene given the provisional designation SrWLR. The BSA identified nine microsatellite (SSR) markers on the long arm of chromosome 2B associated with the resistant phenotype. Fifteen polymorphic SSRs, including the nine identified in the BSA, were used to produce a linkage map of chromosome 2B, positioning SrWLR in an 8.8 cM region between the SSRs GWM47 and WMC332. This region has been reported to contain the wheat stem rust resistance genes Sr9 and SrWeb, the latter conferring resistance to Pgt race TTKSK. The 9,000 marker Illumina Infinium iSelect SNP assay was used to further saturate the SrWLR region. The cosegregating SNP markers IWA6121, IWA6122, IWA7620, IWA8295, and IWA8362 further delimited the SrWLR region distally to a 1.9 cM region. The present study demonstrates the iSelect assay to be an efficient tool to delimit the region of a mapping population and establish syntenic relationships between closely related species.

Using association and linkage mapping, two SNP markers closely linked to the SBWMV resistance gene on chromosome 5D were identified and can be used to select the gene in breeding. Soil-borne wheat mosaic virus (SBWMV) disease is a serious viral disease of winter wheat growing areas worldwide. SBWMV infection can significantly reduce grain yield up to 80 %. Developing resistant wheat cultivars is the only feasible strategy to reduce the losses. In this study, wheat Infinium iSelect Beadchips with 9 K wheat SNPs were used to genotype an association mapping population of 205 wheat accessions. Six new SNPs from two genes were identified to be significantly associated with the gene for SBWMV resistance on chromosome 5D. The SNPs and Xgwm469, an SSR marker that has been reported to be associated with the gene, were mapped close to the gene using F6-derived recombinant inbred lines from the cross between a resistant parent ‘Heyne’ and a susceptible parent ‘Trego’. Two representative SNPs, wsnp_CAP11_c209_198467 and wsnp_JD_c4438_5568170, from the two linked genes in wheat were converted into KBioscience Competitive Allele-Specific Polymerase assays and can be easily used in marker-assisted selection to improve wheat resistance to SBWMV in breeding.

Wheat lines carrying Ug99-effective stem rust resistance gene Sr43 on shortened alien chromosome segments were produced using chromosome engineering, and molecular markers linked to Sr43 were identified for marker-assisted selection. Stem rust resistance gene Sr43, transferred into common wheat (Triticum aestivum) from Thinopyrum ponticum, is an effective gene against stem rust Ug99 races. However, this gene has not been used in wheat breeding because it is located on a large Th. ponticum 7el(2) chromosome segment, which also harbors genes for undesirable traits. The objective of this study was to eliminate excessive Th. ponticum chromatin surrounding Sr43 to make it usable in wheat breeding. The two original translocation lines KS10-2 and KS24-1 carrying Sr43 were first analyzed using simple sequence repeat (SSR) markers and florescent genomic in situ hybridization. Six SSR markers located on wheat chromosome arm 7DL were identified to be associated with the Th. ponticum chromatin in KS10-2 and KS24-1. The results confirmed that KS24-1 is a 7DS·7el(2)L Robertsonian translocation as previously reported. However, KS10-2, which was previously designated as a 7el(2)S·7el(2)L-7DL translocation, was identified as a 7DS-7el(2)S·7el(2)L translocation. To reduce the Th. ponticum chromatin carrying Sr43, a BC(2)F(1) population (Chinese Spring//Chinese Spring ph1bph1b*2/KS10-2) containing ph1b-induced homoeologous recombinants was developed, tested with stem rust, and genotyped with the six SSR markers identified above. Two new wheat lines (RWG33 and RWG34) carrying Sr43 on shortened alien chromosome segments (about 17.5 and 13.7 % of the translocation chromosomes, respectively) were obtained, and two molecular markers linked to Sr43 in these lines were identified. The new wheat lines with Sr43 and the closely linked markers provide new resources for improving resistance to Ug99 and other races of stem rust in wheat.

Wheat (Triticum aestivum L.) flour functionality during the dough development and baking processes is an important quality attribute considered by the baking industry. A flour with high water absorption during mixing means more water and less flour is needed, compared to a flour with less water absorption. The objective of this study was to identify quantitative trait loci (QTL) influencing water absorption and dough rheological properties of hard red spring wheat. QTL were mapped on a genetic linkage map that comprised 531 simple sequence repeats (SSRs) and diversity array technology (DArT) marker loci. Composite interval mapping with 139 recombinant inbred lines (RILs) was used to identify QTL within and across two field environments. Six QTL on chromosomes 1A, 1B, 4B, 4D, and 5A were detected for farinograph water absorption. These QTL also confirmed earlier studies that flour water absorption is a function of protein content, starch damage, and gluten strength. In this study, dough rheological properties such as dough development time, dough stability, mixing tolerance index, and time to breakdown were influenced by the high-molecular weight glutenin genes Glu-B1 and Glu-D1.

Thousand-kernel weight (TKW) is one of the major components of grain yield in wheat (Triticum aestivum). Identifying major quantitative trait loci (QTLs) for TKW and developing effective markers are prerequisite for success in marker-assisted selection (MAS) to improve wheat yield through breeding. This study mapped a major QTL, designated as TaTKW-7AL, for increasing TKW on the long arm of chromosome 7A of ‘Clark’ to a 1.3-cM interval between single nucleotide polymorphism (SNP) markers IWB13913 and IWA5913. This QTL explained 19.7 % of the phenotypic variation for TKW. A QTL for increasing kernel length (KL), one of the major components of TKW, was mapped in the same interval as TaTKW-7AL, suggesting that increased TKW by the QTL in ‘Clark’ is most likely due to the increased KL. Association analysis on a diversity panel of 200 US winter wheat accessions also identified a haplotype of three SNP markers (IWB13913, IWB6693 and IWA5913) that were tightly associated with the both KL and TKW. The analysis of allele frequencies of the haplotype in the diversity panel suggested that the favorable allele of TaTKW-7AL has not been strongly selected for in practice and has potential to be used to improve grain yield in US hard winter wheat breeding. Two user-friendly flanking KASPar markers, IWB13913 and IWA5913, were developed for MAS of TaTKW-7AL.

Wheat stem rust caused by Puccinia graminis f. sp. tritici can cause devastating yield losses in wheat. Over the past several decades, stem rust has been controlled worldwide through the use of genetic resistance. Stem rust race TTKSK (Ug99), first detected in Uganda in 1998, threatens global wheat production because of its unique virulence combination. As the majority of the currently grown cultivars and advanced breeding lines are susceptible to race TTKSK, sources of resistance need to be identified and characterized to facilitate their use in agriculture. South Dakota breeding line SD 1691 displayed resistance to race TTKSK in the international wheat stem rust nursery in Njoro, Kenya. Seedling screening of progeny derived from SD 1691 crossed to susceptible LMPG-6 indicated that a single resistance gene was present. Allelism and race-specificity tests indicated the stem rust resistance gene in SD 1691 was Sr28. The chromosome arm location of Sr28 was previously demonstrated to be 2BL. We identified molecular markers linked to Sr28 and validated this linkage in two additional populations. Common spring wheat cultivars in the central United States displayed allelic diversity for markers flanking Sr28. These markers could be used to select for Sr28 in breeding populations and for combining Sr28 with other stem rust resistance genes.

Much effort has been invested in identifying molecular markers in wheat (Triticum aestivum L.) linked to quantitative trait loci (QTL) that confer resistance to Fusarium head blight (FHB) caused by Fusarium graminearum Schwabe [teleomorph Gibberella zeae (Schwein) Petch]. Even after several generations of crossing and selection by many wheat breeding programs, resistance of the Chinese spring wheat cultivar ‘Sumai 3’ (PI 481542) remains among the most effective. It therefore seems that undocumented resistance QTL present in Sumai 3 were not detected in various mapping studies. Using an extremely susceptible Tibetan landrace (‘Y1193-6’; unknown pedigree) in the creation of a mapping population with Sumai 3, the objective of this research was to identify undocumented resistance QTL in Sumai 3. This was accomplished through collecting disease index (DI) and Fusarium damaged kernel (FDK) phenotypic values along with 305 Diversity Array Technology (DArT) and 52 Simple Sequence Repeat (SSR) marker genotypes on 160 F2:6 recombinant inbred lines (RILs). Disease response evaluations were based on four (two greenhouse and two field) experiments where spray inoculation methods were used. Three QTL were identified on chromosome arms 3BS, 6BL and 2DS explaining 26.1, 10.7 and 18.9% of the phenotypic variation for DI, respectively. The same QTL were also significantly associated with reduced FDK scores and explained 28.0, 11.0 and 23.0% of phenotypic variation. Lines within the mapping population were placed in eight categories with respect to their various QTL combinations. Lines with no QTL were the most susceptible, whereas those with the Sumai 3-derived 3BS and 6BL QTL combined with the 2DS QTL from Y1193-6 were the most resistant. Though the 3BS and 6BL QTL are well-documented, the 2DS resistance QTL, which was contributed by the susceptible parent, confers increased susceptibility when derived from Sumai 3. In this study no new FHB QTL from Sumai 3 was discovered, but results suggest that Sumai 3 contains a QTL for susceptibility on chromosome arm 2DS. Selection against this QTL may potentially increase resistance levels among Sumai 3-derived populations.

Fusarium head blight (FHB) is a devastating disease of wheat worldwide. Novel sources of resistance are critical for improving FHB resistance levels in wheat. From a large-scale evaluation of germplasm for reactions to FHB, we identified one wheat accession (PI 277012) that consistently showed a high level of resistance in both greenhouse and field experiments. To characterize the FHB resistance in this accession, we developed a doubled haploid (DH) mapping population consisting of 130 lines from the cross between PI 277012 and the hard red spring wheat cultivar 'Grandin'. The DH population was then evaluated for reactions to FHB in three greenhouse seasons and five field environments. Based on a linkage map that consisted of 340 SSR markers spanning 2,703 cM of genetic distance, two major quantitative trait loci (QTLs) for FHB resistance were identified on chromosome arms 5AS and 5AL, with each explaining up to 20 and 32% of the variation in FHB severity, respectively. The two QTLs also showed major effects on reducing the percentage of Fusarium damaged kernels (FDK) and deoxynivalenol (DON) accumulation in seeds. FHB resistance has not previously been reported to be associated with this particular genomic region of chromosome arm 5AL, thus indicating the novelty of FHB resistance in PI 277012. Plant maturity was not associated with FHB resistance and the effects of plant height on FHB resistance were minor. Therefore, these results suggest that PI 277012 is an excellent source for improving FHB resistance in wheat. The markers identified in this research are being used for marker-assisted introgression of the QTLs into adapted durum and hard red spring wheat cultivars.

Author(s): Chen, Shisheng; Guo, Yan; Briggs, Jordan; Dubach, Felix; Chao, Shiaoman; Zhang, Wenjun; Rouse, Matthew N; Dubcovsky, Jorge | Abstract: In the original publication, the IWGSC assembly is incorrectly referenced.

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. © 2010 US Government.

Utilization of quantitative trait loci (QTL) identified in bi-parental mapping populations has had limited success for improving complex quantitative traits with low to moderate heritability. Association mapping in contemporary breeding germplasm may lead to more effective marker strategies for crop improvement. To test this approach, we conducted association mapping of two complex traits with moderate heritability; Fusarium head blight (FHB) severity and the grain concentration of mycotoxin associated with disease, deoxynivalenol (DON). To map FHB resistance in barley, 768 breeding lines were evaluated in 2006 and 2007 in four locations. All lines were genotyped with 1,536 SNP markers and QTL were mapped using a mixed model that accounts for relatedness among lines. Average linkage disequilibrium within the breeding germplasm extended beyond 4 cM. Four QTL were identified for FHB severity and eight QTL were identified for the DON concentration in two independent sets of breeding lines. The QTL effects were small, explaining 1–3% of the phenotypic variation, as might be expected for complex polygenic traits. We show that using breeding germplasm to map QTL can complement bi-parental mapping studies by providing independent validation, mapping QTL with more precision, resolving questions of linkage and pleiotropy, and identifying genetic markers that can be applied immediately in crop improvement.

Kernel characteristics, particularly kernel weight, kernel size, and grain protein content, are important components of grain yield and quality in wheat. Development of high performing wheat cultivars, with high grain yield and quality, is a major focus in wheat breeding programs worldwide. Here, we report chromosome regions harboring genes that influence kernel weight, kernel diameter, kernel size distribution, grain protein content, and grain yield in hard red spring wheat breeding lines adapted to the Upper Midwest region of the United States. A genetic linkage map composed of 531 SSR and DArT marker loci spanned a distance of 2,505 cM, covering all 21 chromosomes of wheat. Stable QTL clusters influencing kernel weight, kernel diameter, and kernel size distribution were identified on chromosomes 2A, 5B, and 7A. Phenotypic variation explained by individual QTL at these clusters varied from 5 to 20% depending on the trait. A QTL region on chromosome 2B confers an undesirable pleiotropic effect or a repulsion linkage between grain yield (LOD = 6.7; R 2 = 18%) and grain protein content (LOD = 6.2; R 2 = 13.3%). However, several grain protein and grain yield QTL independent of each other were also identified. Because some of the QTL identified in this study were consistent across environments, DNA markers will provide an opportunity for increasing the frequency of desirable alleles through marker-assisted selection.

Durum wheat (Triticum turgidum L. subsp. durum, 2n = 4x = 28, AABB) is an important cereal used for making pasta products. Compared with bread wheat, durum wheat receives less attention in genetic and genomic studies. In this research, a tetraploid wheat doubled haploid (DH) population derived from the cross between the durum wheat cultivar ‘Lebsock’ and the T. turgidum subsp. carthlicum (2n = 4x = 28, AABB) accession PI 94749 was developed. The population consisted of 146 lines and was used to construct linkage maps of all 14 chromosomes. The maps consisted of 280 SSR markers and spanned 2,034.1 cM with an average density of one marker per 7.2 cM. The DH population and the whole genome linkage maps were then used to identify QTLs associated with tan spot resistance. The DH population was inoculated separately with two Ptr ToxA-producing isolates (Pti2 and 86-124) representing races 1 and 2, respectively, of Pyrenophora tritici-repentis, and five resistance QTLs were detected on chromosome arms 3AS, 3BL, 5AL and 7BL. Together, the QTLs explained a total of 46 and 41% of the phenotypic variation for reaction to Pti2 and 86-124, respectively. The Tsn1-Ptr ToxA interaction was not a significant factor in tan spot development in this population, and none of the QTLs corresponded to previously identified loci known to confer insensitivity to host-selective toxins (HSTs) produced by P. tritici-repentis. This result, together with those of other similar studies, indicates that the wheat–P. tritici-repentis pathosystem involves more factors than currently published host-toxin interactions. The DH population and genetic maps reported here will be useful for genetic dissection of important agronomic traits as well as the identification and development of markers for marker-assisted selection (MAS).

Net type net blotch (NTNB), caused by Pyrenophora teres f. teres Drechs., is prevalent in barley growing regions worldwide. A population of 118 doubled haploid (DH) lines developed from a cross between barley cultivars 'Rika' and 'Kombar' were used to evaluate resistance to NTNB due to their differential reaction to various isolates of P. teres f. teres. Rika was resistant to P. teres f. teres isolate 15A and susceptible to isolate 6A. Conversely, Kombar was resistant to 6A, but susceptible to 15A. A progeny isolate of a 15A x 6A cross identified as 15A x 6A#4 was virulent on both parental lines. The Rika/Kombar (RK) DH population was evaluated for disease reactions to the three isolates. Isolate 15A induced a resistant:susceptible ratio of 78:40 (R:S) whereas isolate 6A induced a resistant:susceptible ratio of 40:78. All but two lines had opposite disease reactions indicating two major resistance genes linked in repulsion. Progeny isolate 15A x 6A#4 showed a resistant:susceptible ratio of 1:117 with the one resistant line also being the single line that was resistant to both 15A and 6A. An RK F(2) population segregated in a 1:3 (R:S) ratio for both 15A and 6A indicating that resistance is recessive. Molecular markers were used to identify a region on chromosome 6H that harbors the two NTNB resistance genes. This work shows that multiple NTNB resistance genes exist at the locus on chromosome 6H, and the recombinant DH line harboring the resistance alleles from both parents will be useful for the development of NTNB-resistant barley germplasm.

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 ofSr42) iseffective 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 uncharacter- ized 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.

Long-range physical maps of the small multigene family of the malt alpha-amylase genes (alpha-Amy-1) located on the long arms of wheat chromosomes 6A (the alpha-Amy-A1 locus) and 6B (alpha-Amy-B1) were generated by pulsed-field gel electrophoresis analysis. By using three methylation-sensitive rare-cutter restriction endonucleases, NotI, NruI and MluI, and an alpha-Amy-1 cDNA probe and four gene-specific genomic probes from the alpha-Amy-B1 locus, the size of the alpha-Amy-A1 locus was estimated to be about 700 kb and of the alpha-Amy-B1 locus to be about approximately 4300 kb. These two maps indicate clustering of GC-rich and C-methylation-sensitive restriction enzyme recognition sites. At least five regions reminiscent of 'CpG islands' are apparent in alpha-Amy-B1, and three in alpha-Amy-A1. Correlation between recombination frequency and physical distance within the alpha-Amy-B1 locus suggests that 1 cM approximates to 1 Mb in physical distance.

Theor Appl Genet (2011) 122:659 DOI 10.1007/s00122-010-1495-2 COMMENT Conflicting mapping results for stem rust resistance gene Sr13 J. Dubcovsky • F. Ordon • D. Perovic B. Admassu • W. Friedt • Z. Abate • W. Zhang • S. Chao Published online: 14 December 2010 O The Author(s) 2010. This article is published with open access at Springerlink.com Two manuscripts with conflicting results on the mapping location of stem rust resistance gene Sr13 in wheat are published in this issue of TAG (Admassu et al. and Simons et al.). Both resistance genes were mapped on the long arm of chromosome 6AL, but the gene mapped by Admassu et al. between markers barc37 and wmc256 is more than 50 cM proximal to the one mapped by Simons et al. between markers CD926040 and BE471213. After the publication of the two manuscripts in TAG online, the authors of both manuscripts became aware of the conflicting results and they exchanged the critical hexaploid wheat cytogenetic stocks Khapstein/9*LMPG (developed by Dr. D. Knott) and the recurrent parent LMPG to deter- mine the source of the differences. In addition, they requested seeds for these two stocks from the U.S. Cereal Disease Laboratory in Saint Paul Minnesota (Dr. Yue Jin) and from Canada Agri-Food, Winnipeg (Dr. Tom Fetch). The online version of the original articles can be found under doi:10.1007/s00122-010-1433-3 and 10.1007/s00122-010-1444-0. J. Dubcovsky (&) Z. Abate W. Zhang Department of Plant Sciences, University of California, Davis, CA 95616, USA e-mail: jdubcovsky@ucdavis.edu F. Ordon D. Perovic B. Admassu Julius Kuehn-Institute, Federal Research Institute for Cultivated Plants (JKI), Institute for Resistance Research and Stress Tolerance, Erwin-Baur-Str. 27, 06484 Quedlinburg, Germany W. Friedt Institute of Crop Science and Plant Breeding I, Justus-Liebig-University Giessen, Heinrich-Buff-Ring 26-32, 35392 Giessen, Germany S. Chao USDA-ARS, Biosciences Research Laboratory, 1605 Albrecht Blvd N, Fargo, ND 58102-2765, USA Analysis of these stocks with molecular markers for the long arm of chromosome 6AL revealed that the Khapstein/ 9*LMPG seed stock used in the Admassu et al. paper was different from the other three Khapstein/9*LMPG stocks. Molecular markers barc37, barc107, barc118, barc1165, gwm570, wmc179, wmc256, wmc580, and cfd2 all showed different bands in the seed stock used in the Admassu paper relative to the seed stocks from the University of Califor- nia, the Cereal Disease Laboratory and Agri-Food Canada. These results indicate that the Khapstein/9*LMPG seed stock used in the Admassu paper is different from the other three. Since the Khapstein/9*LMPG hexaploid genetic stock defines the gene name Sr13, the gene mapped by Admassu et al. is most likely not Sr13 and should be renamed. The Khapstein/9*LMPG developed by Dr. D. Knott is polymorphic with the LMPG recurrent parent only for markers closely linked to the stem rust resistance gene mapped in the four tetraploid populations in the distal region of chromosome arm 6AL (barc104b and dupw167) by Simons et al. This observation, together with the fact that Sr13 was originated in tetraploid wheat (Khapli), provides indirect evidence that the gene mapped by Simons et al. is Sr13. Mapping the stem rust resistance gene present in the correct Khapstein/9*LMPG stock in an hexaploid segregating population will provide the final answer. Interestingly, both genes confer resistance to the new stem rust race Ug99 (TTKSK) suggesting that it would be possible to combine the two sources of resistance to Ug99 in the same 6AL arm. We have initiated crosses between the correct Khapstein/9*LMPG genetic stock and the source of the Admassu et al. Ug99 resistance to recombine both genes. Open Access This article is distributed under the terms of the Creative Commons Attribution Noncommercial License which per- mits any noncommercial use, distribution, and reproduction in any medium, provided the original author(s) and source are credited.

Homologous probes for the wheat coding sequences of the enzymes phosphoribulokinase, phosphoglycerate kinase (both chloroplast and cytosolic forms), chloroplast fructose-1,6-bisphosphatase and the small subunit of ribulose-1,5-bisphosphate carboxylase were used to determine the copy number and chromosomal location of the genes encoding these enzymes by restriction fragment length polymorphism analysis. Heterologous probes were similarly used to characterize the genes for the enzymes glyceraldehyde phosphate dehydrogenase (both chloroplast and cytosolic forms), phosphoenolpyruvate carboxylase and pyruvate, orthophosphate dikinase. Several of the genes are present in single copies per haploid genome, and the different enzymes are encoded by loci dispersed on different chromosomes. The significance of these findings is discussed in relation to gene expression and control of copy number.