Leaf rust caused by
Erikss. (
) and stem rust caused by
f. sp.
i Erikss. & E. Henn (
) are serious constraints to production of durum wheat (
L). The objective of this study was to identify leaf rust ...resistance (
) and stem rust resistance (
) genes/QTL in Portuguese durum landrace PI 192051. Four
-isolates, representing three virulence phenotypes (BBBQJ, BBBSJ & EEEEE) and six
-races TTKSK, JRCQC, TKTTF, QFCFC, TPMKC and TMLKC were used to evaluate 180 recombinant inbred lines (RILs) derived from the cross Rusty (rust susceptible) × PI 192051-1 (rust resistant) at the seedling stage. The RILs were also phenotyped at the adult-plant stage in a stem rust nursery in Ethiopia in 2017. The RILs were genotyped using the Illumina iSelect 9K wheat SNP array. PI 192051-1 carries a previously unidentified major
gene designated as
on chromosome arm 7AS and
gene
in the pericentromeric region of chromosome 4A. In addition, three minor
QTL
,
and
were mapped in PI 192051-1 on chromosomes 1AL, 2BL, and 4A, respectively
could be co-located or tightly linked to
Markers linked to the identified QTL/genes can be used for marker assisted selection. These findings enrich the genetic basis of rust resistance in both durum and common wheat.
Most rust resistance genes thus far isolated from wheat have a very limited number of functional alleles. Here, we report the isolation of most of the alleles at wheat stem rust resistance gene locus ...SR9. The seven previously reported resistance alleles (Sr9a, Sr9b, Sr9d, Sr9e, Sr9f, Sr9g, and Sr9h) are characterised using a synergistic strategy. Loss-of-function mutants and/or transgenic complementation are used to confirm Sr9b, two haplotypes of Sr9e (Sr9e_h1 and Sr9e_h2), Sr9g, and Sr9h. Each allele encodes a highly related nucleotide-binding site leucine-rich repeat (NB-LRR) type immune receptor, containing an unusual long LRR domain, that confers resistance to a unique spectrum of isolates of the wheat stem rust pathogen. The only SR9 protein effective against stem rust pathogen race TTKSK (Ug99), SR9H, differs from SR9B by a single amino acid. SR9B and SR9G resistance proteins are also distinguished by only a single amino acid. The SR9 allelic series found in the B subgenome are orthologs of wheat stem rust resistance gene Sr21 located in the A subgenome with around 85% identity in protein sequences. Together, our results show that functional diversification of allelic variants at the SR9 locus involves single and multiple amino acid changes that recognize isolates of wheat stem rust.
The winter wheat (Triticum aestivum L.) line CI13227 has been characterized as having adult plant resistance to leaf rust caused by Puccinia triticina Eriks (Pt). Line CI13227 was crossed with the ...susceptible spring wheat ‘Thatcher’ (Tc) and a Tc*2/CI13227 F6 line with adult plant leaf rust resistance designated as 411A was derived. Line 411A was crossed with Tc to develop an F6 recombinant inbred line (RIL) population. The parents and 120 F6 lines were assessed for leaf rust severity at the flag leaf stage in five field plot tests from 2011 through 2015 and were genotyped for single‐nucleotide polymorphism (SNP) markers with the Illumina iSelect 90K wheat bead array. A total of 2,384 SNP markers segregated among the RILs. Completely linked SNPs were removed, and 474 markers that covered 2,605 centimorgans (cM) were used for linkage map construction. Quantitative trait loci (QTL) on chromosome 2AL with logarithm of odds (LOD) values 2.34–7.88, on chromosome 4BS with LOD values 1.35– 4.66, and on chromosome 7AL with LOD values 2.92–7.81 were associated with significant reduction in leaf rust severity in the field plot tests. Recombinant inbred lines that had combinations of two or three of the QTL had significantly lower leaf rust severity than RILs that lacked any resistance QTL. Kompetitive allele specific polymerase chain reaction (KASP) markers were developed for the SNPs that were closely linked with the three QTL to facilitate marker‐based selection of the leaf rust resistance in breeding programs.
Core Ideas
Winter wheat line CI13227 has adult plant leaf rust resistance.
QTL for adult plant leaf rust resistance from CI13227 were mapped to chromosomes 2AL, 4BS, and 7AL.
KASP markers linked to the QTL regions were developed.
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FZAB, GIS, IJS, KILJ, NLZOH, NUK, OILJ, SAZU, SBCE, SBMB, UL, UM, UPUK
Key message
The diploid wheat recessive stem rust resistance gene
SrTm4
was fine-mapped to a 754-kb region on chromosome arm 2A
m
L and potential candidate genes were identified.
Race Ug99 of
...Puccinia graminis
f. sp.
tritici
(
Pgt
), the causal agent of wheat stem (or black) rust is one of the most serious threats to global wheat production. The identification, mapping, and deployment of effective stem rust resistance (
Sr
) genes are critical to reduce this threat. In this study, we generated
SrTm4
monogenic lines and found that this gene confers resistance to North American and Chinese
Pgt
races. Using a large mapping population (9522 gametes), we mapped
SrTm4
within a 0.06 cM interval flanked by marker loci
CS4211
and
130K1519
, which corresponds to a 1.0-Mb region in the Chinese Spring reference genome v2.1. A physical map of the
SrTm4
region was constructed with 11 overlapping BACs from the resistant
Triticum monococcum
PI 306540. Comparison of the 754-kb physical map with the genomic sequence of Chinese Spring and a discontinuous BAC sequence of DV92 revealed a 593-kb chromosomal inversion in PI 306540. Within the candidate region, we identified an L-type lectin-domain containing receptor kinase (
LLK1
), which was disrupted by the proximal inversion breakpoint, as a potential candidate gene. Two diagnostic dominant markers were developed to detect the inversion breakpoints. In a survey of
T. monococcum
accessions, we identified 10 domesticated
T. monococcum
subsp.
monococcum
genotypes, mainly from the Balkans, carrying the inversion and showing similar mesothetic resistant infection types against
Pgt
races. The high-density map and tightly linked molecular markers developed in this study are useful tools to accelerate the deployment of
SrTm4
-mediated resistance in wheat breeding programs.
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EMUNI, FIS, FZAB, GEOZS, GIS, IJS, IMTLJ, KILJ, KISLJ, MFDPS, NLZOH, NUK, OILJ, PNG, SAZU, SBCE, SBJE, SBMB, SBNM, UKNU, UL, UM, UPUK, VKSCE, ZAGLJ
Summary
Wheat stem (or black) rust, caused by Puccinia graminis f. sp. tritici (Pgt), has been historically among the most devastating global fungal diseases of wheat. The recent occurrence and ...spread of new virulent races such as Ug99 have prompted global efforts to identify and isolate more effective stem rust resistance (Sr) genes. Here, we report the map‐based cloning of the Ug99‐effective SrTm5 gene from diploid wheat Triticum monococcum accession PI 306540 that encodes a typical coiled‐coil nucleotide‐binding leucine‐rich repeat protein. This gene, designated as Sr22b, is a new allele of Sr22 with a rare insertion of a large (13.8‐kb) retrotransposon into its second intron. Biolistic transformation of an ~112‐kb circular bacterial artificial chromosome plasmid carrying Sr22b into the susceptible wheat variety Fielder was sufficient to confer resistance to stem rust. In a survey of 168 wheat genotypes, Sr22b was present only in cultivated T. monococcum subsp. monococcum accessions but absent in all tested tetraploid and hexaploid wheat lines. We developed a diagnostic molecular marker for Sr22b and successfully introgressed a T. monococcum chromosome segment containing this gene into hexaploid wheat to accelerate its deployment and pyramiding with other Sr genes in wheat breeding programmes. Sr22b can be a valuable component of gene pyramids or transgenic cassettes combining different resistance genes to control this devastating disease.
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BFBNIB, DOBA, FZAB, GIS, IJS, IZUM, KILJ, NLZOH, NUK, OILJ, PILJ, PNG, SAZU, SBCE, SBMB, UILJ, UKNU, UL, UM, UPUK
Wheat stem rust caused by
f. sp.
threatens wheat production worldwide. The objective of this study was to characterize wheat stem rust resistance in 'Linkert', a variety with adult plant resistance ...effective to emerging wheat stem rust pathogen strain Ug99. Two doubled haploid (DH) populations and one recombinant inbred line (RIL) population were developed with 'Linkert' as a stem rust resistant parent. Hard red spring wheat variety 'Forefront' and genetic stock 'LMPG' were used as stem rust susceptible parents of the DH populations. Breeding line 'MN07098-6' was used as a susceptible parent of the RIL population. Both DH and RIL populations with their parents were evaluated both at the seedling stage and in the field against
races. Genotyping data of the DH populations were generated using the wheat iSelect 90k SNP assay. The RIL population was genotyped by genotyping-by-sequencing. We found QTL consistently associated with wheat stem rust resistance on chromosome 2BS for the Linkert/Forefront DH population and the Linkert/MN07098-6 RIL population both in Ethiopia and Kenya. Additional reliable QTL were detected on chromosomes 5BL (125.91 cM) and 4AL (
) for the Linkert/LMPG population in Ethiopia and Kenya. Different QTL identified in the populations reflect the importance of examining the genetics of resistance in populations derived from adapted germplasm (Forefront and MN07098-6) in addition to a genetic stock (LMPG). The associated markers in this study could be used to track and select for the identified QTL in wheat breeding programs.
The identification and characterization of resistance genes should outpace the rapid emergence of new P. graminis f. sp. tritici races, such as TTRTF and TTKTT, to mitigate stem rust damage to wheat. ...The objective of the current study was to identify and characterize P. graminis f. sp. tritici race resistance association signals. A total of 250 North American spring wheat lines were evaluated at the seedling stage with a total of seven isolates including TKKTP, TKTTF, TKTTF, TRTTF, TTRTF, TTKSK, and TTKTT. The lines were genotyped by a GBS platform and 9,042 SNPs were used for identification of chromosome regions associated with resistance against the seven isolates. Strong association signals were detected on chromosomes 6BL (Sr11 gene region) and 4AL, likely Sr7a, for resistance against both TKKTP and TKTTF. Similarly, association signals were also detected on chromosomes 4AL (race TTRTF resistance) and 4BS (race TTKSK and TTKTT resistance). Association analysis based on mean phenotypic differences between closely related isolates identified QTL that were not elucidated by direct association mapping of the responses, individually. Overall, with the exception of race TRTTF, each race shared at least one association signal with another race. However, the number of race‐specific association signals are larger than that of association signals common among races suggesting the need for identifying and characterizing QTL/genes for newly emerging stem rust pathogen races. There was also high concordance between PCA‐based GWAS association signals and association signals from that of both single and multi‐locus mixed models.
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FZAB, GIS, IJS, KILJ, NLZOH, NUK, OILJ, SAZU, SBCE, SBMB, UL, UM, UPUK
Leaf rust, caused by
(
), stripe rust caused by
f. sp.
(
), and stem rust caused by
f. sp.
(
) are major diseases to wheat production globally. Host resistance is the most suitable approach to manage ...these fungal pathogens. We investigated the phenotypic and genotypic structure of resistance to leaf rust, stem rust, and stripe rust pathogen races at the seedling stage in a collection of advanced durum wheat breeding lines and cultivars adapted to Upper Mid-West region of the United States. Phenotypic evaluation showed that the majority of the durum wheat genotypes were susceptible to
isolates adapted to durum wheat, whereas all the genotypes were resistant to common wheat type-
isolate. The majority of genotypes were resistant to stripe rust and stem rust pathogen races. The durum panel genotyped using Illumina iSelect 90 K wheat SNP assay was used for genome-wide association mapping (GWAS). The GWAS revealed 64 marker-trait associations (MTAs) representing six leaf rust resistance loci located on chromosome arms 2AS, 2AL, 5BS, 6AL, and 6BL. Two of these loci were identified at the positions of
and
genes, whereas the remaining loci are most likely novel. A total of 46 MTAs corresponding to four loci located on chromosome arms 1BS, 5BL, and 7BL were associated with stripe rust response. None of these loci correspond to designated stripe rust resistance genes. For stem rust, a total of 260 MTAs, representing 22 loci were identified on chromosome arms 1BL, 2BL, 3AL, 3BL, 4AL, 5AL, 5BL, 6AS, 6AL, 6BL, and 7BL. Four of these loci were located at the positions of known genes/alleles (
,
,
, and
). The discovery of known and novel rust resistance genes and their linked SNPs will help diversify rust resistance in durum wheat.
ABSTRACT
Array‐ or chip‐based single nucleotide polymorphism (SNP) markers are widely used in genomic studies because of their abundance in a genome and lower cost per data point than older marker ...technologies. Genotyping‐by‐sequencing (GBS), a relatively newer approach of genotyping, suggests equal appeal because of its lesser cost per data point and the avoidance of ascertainment bias during genotyping. In this study, we compared the results from quantitative trait loci (QTL) mapping, marker distribution on linkage maps, genome size, recombination sites covered by the markers, and cost per polymorphic marker, as well as the methodology and workflow between the Illumina Infinium 9000 SNP‐chip genotyping with GBS. Results indicate that while GBS offers similar genome coverage at almost one‐fourth the cost of SNP chip, the SNP‐chip method is less demanding of computational skills and resources. Eight and nine QTL were detected in the GBS and SNP‐chip datasets, respectively, with one QTL common between the systems. Additionally, imputation accuracy of the GBS dataset was examined by introducing missing values randomly and imputing the missing alleles using a probabilistic principal components algorithm. Imputation results suggest recovery of the missing alleles with reasonable accuracy in datasets with low (up to 40%) amount of missing data is possible and can provide acceptable accuracy in gene mapping. Overall, the comparative results indicate that both approaches provide good genome coverage and similar mapping results. The choice of the genotyping platform is decided by the nature of the study and available resources.
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FZAB, GIS, IJS, KILJ, NLZOH, NUK, OILJ, SAZU, SBCE, SBMB, UL, UM, UPUK
The narrow genetic basis of resistance in modern wheat cultivars and the strong selection response of pathogen populations have been responsible for periodic and devastating epidemics of the wheat ...rust diseases. Characterizing new sources of resistance and incorporating multiple genes into elite cultivars is the most widely accepted current mechanism to achieve durable varietal performance against changes in pathogen virulence. Here, we report a high-density molecular characterization and genome-wide association study (GWAS) of stripe rust and stem rust resistance in 190 Ethiopian bread wheat lines based on phenotypic data from multi-environment field trials and seedling resistance screening experiments. A total of 24,281 single nucleotide polymorphism (SNP) markers filtered from the wheat 90 K iSelect genotyping assay was used to survey Ethiopian germplasm for population structure, genetic diversity and marker-trait associations.
Upon screening for field resistance to stripe rust in the Pacific Northwest of the United States and Ethiopia over multiple growing seasons, and against multiple races of stripe rust and stem rust at seedling stage, eight accessions displayed resistance to all tested races of stem rust and field resistance to stripe rust in all environments. Our GWAS results show 15 loci were significantly associated with seedling and adult plant resistance to stripe rust at false discovery rate (FDR)-adjusted probability (P) <0.10. GWAS also detected 9 additional genomic regions significantly associated (FDR-adjusted P < 0.10) with seedling resistance to stem rust in the Ethiopian wheat accessions. Many of the identified resistance loci were mapped close to previously identified rust resistance genes; however, three loci on the short arms of chromosomes 5A and 7B for stripe rust resistance and two on chromosomes 3B and 7B for stem rust resistance may be novel.
Our results demonstrate that considerable genetic variation resides within the landrace accessions that can be utilized to broaden the genetic base of rust resistance in wheat breeding germplasm. The molecular markers identified in this study should be useful in efficiently targeting the associated resistance loci in marker-assisted breeding for rust resistance in Ethiopia and other countries.
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DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK