Evaluation of Pakistan wheat germplasms for stripe rust resistance using molecular markers

Wheat production in Pakistan is seriously constrained due to rust diseases and stripe rust (yellow) caused by Puccinia striiformis f. sp. tritici, which could limit yields. Thus development and cultivation of genetically diverse and resistant varieties is the most sustainable solution to overcome these diseases. The first objective of the present study was to evaluate 100 Pakistan wheat cultivars that have been grown over the past 60 years. These cultivars were inoculated at the seedling stage with two virulent stripe rust isolates from the United States and two from Pakistan. None of the wheat cultivars were resistant to all tested stripe rust isolates, and 16% of cultivars were susceptible to the four isolates at the seedling stage. The data indicated that none of the Pakistan wheat cultivars contained either Yr5 or Yr15 genes that were considered to be effective against most P. striiformis f. sp. tritici isolates from around the world. Several Pakistan wheat cultivars may have gene Yr10, which is effective against isolate PST-127 but ineffective against PST-116. It is also possible that these cultivars may have other previously unidentified genes or gene combinations. The second objective was to evaluate the 100 Pakistan wheat cultivars for stripe rust resistance during natural epidemics in Pakistan and Washington State, USA. It was found that a higher frequency of resistance was present under field conditions compared with greenhouse conditions. Thirty genotypes (30% of germplasms) were found to have a potentially high temperature adult plant (HTAP) resistance. The third objective was to determine the genetic diversity in Pakistan wheat germplasms using molecular markers. This study was based on DNA fingerprinting using resistance gene analog polymorphism (RGAP) marker analysis. The highest polymorphism detected with RGAP primer pairs was 40%, 50% and 57% with a mean polymorphism of 36%. A total of 22 RGAP markers were obtained in this study. RGAP, simple sequence repeat (SSR) and sequence tagged site (STS) markers were used to determine the presence and absence of some important stripe rust resistance genes, such as Yr5, Yr8, Yr9, Yr15 and Yr18. Of the 60 cultivars analyzed, 17% of cultivars showed a RGAP marker band for Yr9 and 12% of cultivars exhibited the Yr18 marker band. No marker band was detected for Yr5, Yr8 and Yr15, indicating a likely absence of these genes in the tested Pakistan wheat cultivars. Cluster analysis based on molecular and stripe rust reaction data is useful in identifying considerable genetic diversity among Pakistan wheat cultivars. The resistant germplasms identified with 22 RGAP markers and from the resistance evaluations should be useful in developing new wheat cultivars with stripe rust resistance.     

Characterization and molecular mapping of stripe rust resistance gene Yr61 in winter wheat cultivar Pindong 34

Key message

We report a new stripe rust resistance gene on chromosome 7AS in wheat and molecular markers useful for transferring it to other wheat genotypes.

Abstract

Several new races of the stripe rust pathogen have established throughout the wheat growing regions of China in recent years. These new races are virulent to most of the designated seedling resistance genes limiting the resistance sources. It is necessary to identify new genes for diversification and for pyramiding different resistance genes in order to achieve more durable resistance. We report here the identification of a new resistance gene, designated as Yr61, in Chinese wheat cultivar Pindong 34. A mapping population of 208 F₂ plants and 128 derived F_2:3 lines in a cross between Mingxian 169 and Pindong 34 was evaluated for seedling stripe rust response. A genetic map consisting of eight resistance gene analog polymorphism (RGAP), two sequence-tagged site (STS) and four simple sequence repeat (SSR) markers was constructed. Yr61 was located on the short arm of chromosome 7A and flanked by RGAP markers Xwgp5467 and Xwgp5765 about 1.9 and 3.9 cM in distance, which were successfully converted into STS markers STS5467 and STS5765b, respectively. The flanking STS markers could be used for marker-assisted selection of Yr61 in breeding programs.     

Molecular mapping of a gene for stripe rust resistance in spring wheat cultivar IDO377s

Stripe rust, caused by Puccinia striiformis f. sp. tritici, is one of the most important diseases of wheat worldwide. The best strategy to control stripe rust is to grow resistant cultivars. One such cultivar resistant to most races in North America is ‘IDO377s’. To study the genetics of its resistance this spring wheat cultivar was crossed with ‘Avocet Susceptible’ (AvS). Seedlings of the parents, F₂ plants, and F₃ lines were tested under controlled greenhouse conditions with races PST-43 and PST-45 of P. striiformis f. sp. tritici. IDO377s carries a single dominant gene for resistance. Resistance gene analog polymorphism (RGAP) and simple sequence repeat (SSR) techniques were used to identify molecular markers linked to the resistance gene. A total of ten markers were identified, two of which flanked the locus at 4.4 and 5.5 cM. These flanking RGAP markers were located on chromosome 2B with nulli-tetrasomic lines of ‘Chinese Spring’. Their presence in the ditelosomic 2BL line localized them to the long arm. The chromosomal location of the resistance gene was further confirmed with two 2BL-specific SSR markers and a sequence tagged site (STS) marker previously mapped to 2BL. Based on the chromosomal location, reactions to various races of the pathogen and tests of allelism, the IDO377s gene is different from all previously designated genes for stripe rust resistance, and is therefore designated Yr43. A total of 108 wheat breeding lines and cultivars with IDO377s or related cultivars in their parentage were assayed to assess the status of the closest flanking markers and to select lines carrying Yr43. The results showed that the flanking markers were reliable for assisting selection of breeding lines carrying the resistance gene. A linked stripe rust resistance gene, previously identified as YrZak, in cultivar Zak was designated Yr44.     

New slow-rusting leaf rust and stripe rust resistance genes <Emphasis Type="Italic">Lr67</Emphasis> and <Emphasis Type="Italic">Yr46</Emphasis> in wheat are pleiotropic or closely linked

The common wheat genotype ‘RL6077’ was believed to carry the gene Lr34/Yr18 that confers slow-rusting adult plant resistance (APR) to leaf rust and stripe rust but located to a different chromosome through inter-chromosomal reciprocal translocation. However, haplotyping using the cloned Lr34/Yr18 diagnostic marker and the complete sequencing of the gene indicated Lr34/Yr18 is absent in RL6077. We crossed RL6077 with the susceptible parent ‘Avocet’ and developed F₃, F₄ and F₆ populations from photoperiod-insensitive F₃ lines that were segregating for resistance to leaf rust and stripe rust. The populations were characterized for leaf rust resistance at two Mexican sites, Cd. Obregon during the 2008–2009 and 2009–2010 crop seasons, and El Batan during 2009, and for stripe rust resistance at Toluca, a third Mexican site, during 2009. The F₃ population was also evaluated for stripe rust resistance at Cobbitty, Australia, during 2009. Most lines had correlated responses to leaf rust and stripe rust, indicating that either the same gene, or closely linked genes, confers resistance to both diseases. Molecular mapping using microsatellites led to the identification of five markers (Xgwm165, Xgwm192, Xcfd71, Xbarc98 and Xcfd23) on chromosome 4DL that are associated with this gene(s), with the closest markers being located at 0.4 cM. In a parallel study in Canada using a Thatcher × RL6077 F₃ population, the same leaf rust resistance gene was designated as Lr67 and mapped to the same chromosomal region. The pleiotropic, or closely linked, gene derived from RL6077 that conferred stripe rust resistance in this study was designated as Yr46. The slow-rusting gene(s) Lr67/Yr46 can be utilized in combination with other slow-rusting genes to develop high levels of durable APR to leaf rust and stripe rust in wheat.     

Genetics and molecular mapping of genes for high-temperature resistance to stripe rust in wheat cultivar Xiaoyan 54

Stripe rust, caused by Puccinia striiformis f. sp. tritici, is one of the most widespread and destructive wheat diseases worldwide. Growing resistant cultivars is the preferred means of control of the disease. The winter wheat cultivar Xiaoyan 54 has high-temperature resistance to stripe rust. To identify genes for stripe rust resistance, Xiaoyan 54 was crossed with Mingxian 169, a winter wheat genotype susceptible to all Chinese races of the pathogen. Seedlings and adult plants of the parents and F₁, F₂, F₃ and F₄ progeny were tested with Chinese race CYR32 under controlled greenhouse conditions and in the field. Xiaoyan 54 has two recessive resistance genes, designated as Yrxy1 and Yrxy2, conferring high-temperature resistance. Simple sequence repeat (SSR) primers were used to identify molecular markers flanking Yrxy2 using 181 plants from one segregating F₃ line. A total of nine markers, two of which flanked the locus at genetic distances of 4.0 and 6.4 cM on the long arm of chromosome 2A were identified. Resistance gene analog polymorphism (RGAP) and SSR techniques were used to identify molecular markers linked to Yrxy1. A linkage group of nine RGAP and two SSR markers was constructed for Yrxy1 using 177 plants of another segregating F₃ line. Two RGAP markers were closely linked to the locus with genetic distances of 2.3 and 3.5 cM. Amplification of a set of nulli-tetrasomic Chinese Spring lines with RGAP markers M8 and M9 and the two SSR markers located Yrxy1 on the short arm of chromosome 7A. The SSR markers Xbarc49 and Xwmc422 were 15.8 and 26.1 cM, respectively, from the gene. The closely linked molecular markers should be useful for incorporating the resistance genes into commercial cultivars and combining them with other genes for stripe rust resistance.     

Development of resistance gene analog polymorphism markers for the Yr9 gene resistance to wheat stripe rust.

The Yr9 gene, which confers resistance to stripe rust caused by Puccinia striiformis f.sp. tritici (P. s. tritici) and originated from rye, is present in many wheat cultivars. To develop molecular markers for Yr9, a Yr9 near-isogenic line, near-isogenic lines with nine other Yr genes, and the recurrent wheat parent 'Avocet Susceptible' were evaluated for resistance in the seedling stage to North American P s. tritici races under controlled temperature in the greenhouse. The resistance gene analog polymorphism (RGAP) technique was used to identify molecular markers for Yr9. The BC7:F, and BC7:F3 progeny, which were developed by backcrossing the Yr9 donor wheat cultivar Clement with 'Avocet Susceptible', were evaluated for resistance to stripe rust races. Genomic DNA was extracted from 203 BC7:F2 plants and used for cosegregation analysis. Of 16 RGAP markers confirmed by cosegregation analysis, 4 were coincident with Yr9 and 12 were closely linked to Yr9 with a genetic distance ranging from 1 to 18 cM. Analyses of nullitetrasomic 'Chinese Spring' lines with the codominant RGAP marker Xwgp13 confirmed that the markers and Yr9 were located on chromosome 1B. Six wheat cultivars reported to have 1B/1R wheat-rye translocations and, presumably, Yr9, and two rye cultivars were inoculated with four races of P. s. tritici and tested with 9 of the 16 RGAP markers. Results of these tests indicate that 'Clement', 'Aurora', 'Lovrin 10', 'Lovrin 13', and 'Riebesel 47/51' have Yr9 and that 'Weique' does not have Yr9. The genetic information and molecular markers obtained from this study should be useful in cloning Yr9, in identifying germplasm that may have Yr9, and in using marker-assisted selection for combining Yr9 with other stripe rust resistance genes.     

<Emphasis Type="Italic">Yr45</Emphasis>, a new wheat gene for stripe rust resistance on the long arm of chromosome 3D

Stripe rust, caused by Puccinia striiformis f. sp. tritici, is one of the most destructive diseases of wheat worldwide. Growing resistant cultivars is the most effective approach to control the disease, but only a few genes confer effective all-stage resistance against the current populations of the pathogen worldwide. It is urgent to identify new genes for diversifying sources of resistance genes and for pyramiding genes for different types of resistance in order to achieve high levels of durable resistance for sustainable control of stripe rust. The common spring wheat genotype ‘PI 181434’, originally from Afghanistan, was resistant in all greenhouse and field tests in our previous studies. To identify the resistance gene(s) PI 181434 was crossed with susceptible genotype ‘Avocet Susceptible’. Adult plants of 103 F₂ progeny were tested in the field under the natural infection of P. striiformis f. sp. tritici. Seedlings of the parents, F₂ and F₃ were tested with races PST-100 and PST-127 of the pathogen under controlled greenhouse conditions. The genetic study showed that PI 181434 has a single dominant gene conferring all-stage resistance. Resistance gene analog polymorphism (RGAP) and simple sequence repeat (SSR) techniques were used to identify molecular markers linked to the gene. A linkage map of 8 RGAP and 2 SSR markers was constructed for the gene using data from the 103 F₂ plants and their derived F₃ lines tested in the greenhouse. Amplification of the complete set of nulli-tetrasomic lines and selected ditelosomic lines of Chinese Spring with an RGAP marker and the two SSR markers mapped the gene on the long arm of chromosome 3D. Because it is the first gene for stripe rust resistance mapped on chromosome 3DL and different from all previously named Yr genes, the gene in PI 181434 was designated Yr45. Polymorphism rates of the two closest flanking markers, Xwgp115 and Xwgp118, in 45 wheat genotypes were 73.3 and 82.2%, respectively. Single nucleotide polymorphisms (SNPs) were identified in the eight wheat genotypes sharing both flanking markers. The RGAP markers and potential SNP markers should be useful in incorporating the gene into wheat cultivars and in pyramiding it with other genes for durable resistance.     

黄淮麦区小麦品种(系)中Yr26基因的SSR检测

选用与Yr26紧密连锁的SSR标记Xgwm11和Xgwm18结合田间抗性鉴定,对239份黄淮麦区小麦品种(系)进行检测,以明确Yr26基因在黄淮麦区小麦品种资源中的分布.结果表明:共有35份品种(系)含有与Yr26紧密连锁的SSR标记Xgwm18或Xgwm11的特征带,占检测样本的14.6%.在这35份材料中,31份田间抗性鉴定表现免疫至中抗,4份表现中感.分子标记检测与田间抗病性检测吻合度较好,该标记可以用于Yr26基因的分子标记辅助选择.综合分子标记和田间鉴定,31份小麦(系)含有Yr26基因,占102份抗病材料的30.39%.     

Genetics and molecular mapping of genes for race-specific all-stage resistance and non-race-specific high-temperature adult-plant resistance to stripe rust in spring wheat cultivar Alpowa

Stripe rust, caused by Puccinia striiformis f. sp. tritici, is one of the most widespread and destructive wheat diseases worldwide. Growing resistant cultivars is the preferred control of the disease. The spring wheat cultivar ‘Alpowa’ has both race-specific, all-stage resistance and non-race-specific, high-temperature adult-plant (HTAP) resistances to stripe rust. To identify genes for the stripe rust resistances, Alpowa was crossed with ‘Avocet Susceptible’ (AVS). Seedlings of the parents, and F₁, F₂ and F₃ progeny were tested with races PST-1 and PST-21 of P. striiformis f. sp. tritici under controlled greenhouse conditions. Alpowa has a single partially dominant gene, designated as YrAlp, conferring all-stage resistance. Resistance gene analog polymorphism (RGAP) and simple sequence repeat (SSR) techniques were used to identify molecular markers linked to YrAlp. A linkage group of five RGAP markers and two SSR markers was constructed for YrAlp using 136 F₃ lines. Amplification of a set of nulli-tetrasomic Chinese Spring lines with RGAP markers Xwgp47 and Xwgp48 and the two SSR markers indicated that YrAlp is located on the short arm of chromosome 1B. To map quantitative trait loci (QTLs) for the non-race-specific HTAP resistance, the parents and 136 F₃ lines were tested at two sites near Pullman and one site near Mount Vernon, Washington, under naturally infected conditions. A major HTAP QTL was consistently detected across environments and was located on chromosome 7BL. Because of its chromosomal location and the non-race-specific nature of the HTAP resistance, this gene is different from previously described genes for adult-plant resistance, and is therefore designated Yr39. The gene contributed to 64.2% of the total variation of relative area under disease progress curve (AUDPC) data and 59.1% of the total variation of infection type data recorded at the heading-flowering stages. Two RGAP markers, Xwgp36 and Xwgp45 with the highest R ² values were closely linked to Yr39, should be useful for incorporation of the non-race-specific resistance gene into new cultivars and for combining Yr39 with other genes for durable and high-level resistance.     

Characterisation of a new stripe rust resistance gene <Emphasis Type="Italic">Yr47</Emphasis> and its genetic association with the leaf rust resistance gene <Emphasis Type="Italic">Lr52</Emphasis>

Two Iranian common wheat landraces AUS28183 and AUS28187 from the Watkins collection showed high levels of seedling resistance against Australian pathotypes of leaf rust and stripe rust pathogens. Chi-squared analyses of rust response segregation among F₃ populations derived from crosses of AUS28183 and AUS28187 with a susceptible genotype AUS27229 revealed monogenic inheritance of leaf rust and stripe rust resistance. As both genotypes produced similar leaf rust and stripe rust infection types, they were assumed to carry the same genes. The genes were temporarily named as LrW1 and YrW1. Molecular mapping placed LrW1 and YrW1 in the short arm of chromosome 5B, about 10 and 15 cM proximal to the SSR marker gwm234, respectively, and the marker cfb309 mapped 8–12 cM proximal to YrW1. LrW1 mapped 3–6 cM distal to YrW1 in two F₃ populations. AUS28183 corresponded to the accession V336 of the Watkins collection which was the original source of Lr52. Based on the genomic location and accession records, LrW1 was concluded to be Lr52. Because no other seedling stripe rust resistance gene has previously been mapped in chromosome 5BS, YrW1 was permanently named as Yr47. A combination of flanking markers gwm234 and cfb309 with phenotypic assays could be used to ascertain the presence of Lr52 and Yr47 in segregating populations. This investigation characterised a valuable source of dual leaf rust and stripe rust resistance for deployment in new wheat cultivars. Transfer of Lr52 and Yr47 into current Australian wheat backgrounds is in progress.     

Molecular Detection of Stripe Rust Resistance Gene(s) in 115 Wheat Cultivars (lines) from the Yellow and Huai River Valley Wheat Region

Stripe rust (yellow rust), caused by Puccinia striiformis f.sp. tritici (Pst), is a serious disease of wheat worldwide, including China. Growing resistant cultivars is the most cost‐effective and environmentally friendly approach to control the disease. To assess the stripe rust resistance in commercial wheat cultivars and advanced lines in the Yellow and Huai River Valley Wheat Region, 115 wheat cultivars (lines) collected from 13 provinces in this region were evaluated with the most prevalent Chinese Pst races CYR32, CYR33 and the new race V26 at seedling stage. In addition, these wheat entries were inoculated with the mixed races of CYR32 and CYR33 at the adult‐plant stage in the field. The results indicated that 53 (46.1%) cultivars (lines) had all‐stage resistance to all the three races, and 16 (13.9%) cultivars (lines) showed adult‐plant resistance. The possible stripe rust resistance genes in these entries were postulated by the closely linked markers of all‐stage resistance genes Yr5, Yr9, Yr10, Yr15 and Yr26 and adult‐plant resistance gene Yr18. Molecular analysis indicated that resistance genes Yr5, Yr9, Yr10, Yr18 and Yr26 were found in 5 (4.3%), 38 (33.0%), 1 (0.9%), 2 (1.7%) and 8 (7.0%) entries, respectively. No entry was found to carry the Yr15 gene. In future breeding programs, Yr5, Yr15 and Yr18 should be used to pyramid with other effective genes to develop wheat cultivars with high‐level and durable resistance to stripe rust, whereas Yr9, Yr10 and Yr26 should not be used or used in a limited way due to the virulent races present in China.     

Molecular mapping of stripe rust resistance gene YrCH42 in Chinese wheat cultivar Chuanmai 42 and its allelism with Yr24 and Yr26

Stripe rust, caused by Puccinia striiformis f. sp. tritici (PST), is one of the most devastating diseases in common wheat (Triticum aestivum L.) worldwide. The objectives of this study were to map a stripe rust resistance gene in Chinese wheat cultivar Chuanmai 42 using molecular markers and to investigate its allelism with Yr24 and Yr26. A total of 787 F₂ plants and 186 F₃ lines derived from a cross between resistant cultivar Chuanmai 42 and susceptible line Taichung 29 were used for resistance gene tagging. Also 197 F₂ plants from the cross Chuanmai 42×Yr24/3*Avocet S and 726 F₂ plants from Chuanmai 42×Yr26/3*Avocet S were employed for allelic test of the resistance genes. In all, 819 pairs of wheat SSR primers were used to test the two parents, as well as resistant and susceptible bulks. Subsequently, nine polymorphic markers were employed for genotyping the F₂ and F₃ populations. Results indicated that the stripe rust resistance in Chuanmai 42 was conferred by a single dominant gene, temporarily designated YrCH42, located close to the centromere of chromosome 1B and flanked by nine SSR markers Xwmc626, Xgwm273, Xgwm11, Xgwm18, Xbarc137, Xbarc187, Xgwm498, Xbarc240 and Xwmc216. The resistance gene was closely linked to Xgwm498 and Xbarc187 with genetic distances of 1.6 and 2.3 cM, respectively. The seedling tests with 26 PST isolates and allelic tests indicated that YrCH42, Yr24 and Yr26 are likely to be the same gene.G.Q. Li and Z.F. Li contributed equally to the work.     

Mapping a stripe rust resistance gene YrC591 in wheat variety C591 with SSR and AFLP markers

Stripe rust, caused by Puccinia striiformis Westend. f. sp. tritici (PST), is one of the most destructive diseases of common wheat (Triticum aestivum L.). To determine inheritance of stripe rust resistance and map the resistance gene(s) in wheat variety C591, F₁, F_2, and F₃ progenies derived from the Taichung 29 × C591 cross were inoculated with Chinese PST race CY32 in the greenhouse. Genetic analysis identified a single dominant gene, temporarily designated YrC591. A total of 178 SSR and 130 AFLP markers were used to test the parents and resistant and susceptible bulks. From the bulk segregant analysis, seven polymorphic SSR and two AFLP markers were selected for genotyping the F₂ population. SSR marker Xcfa2040-7B, and SCAR marker SC-P35M48 derived from AFLP marker P35M48 ₃₇₃ were identified to be closely linked to the resistance gene with genetic distances of 8.0 and 11.7 cM, respectively. The SSR markers mapped the resistance gene on chromosome arm 7BL. In the seedling test with five PST races, the reaction patterns of C591 were different from wheat cultivars or lines carrying Yr2 or Yr6 that also are found on chromosome 7B. The results indicate that YrC591 is probably a novel stripe rust resistance gene.     

Molecular tagging of stripe rust resistance gene YrZH84 in Chinese wheat line Zhou 8425B

Stripe rust, caused by Puccinia striiformis f. sp. tritici (PST), is one of the most damaging diseases in common wheat (Triticum aestivum L.). With the objective of identifying and tagging new genes for resistance to stripe rust, F₁, F₂ and F₃ populations from the cross Zhou 8425B/Chinese Spring were inoculated with Chinese PST isolate CYR32 in the greenhouse. A total of 790 SSR primers were used to test the parents and resistant and susceptible bulks. The resulting seven polymorphic markers on chromosome 7BL were used for genotyping F₂ and F₃ populations. Results indicated that Zhou 8425B carries a single dominant resistance gene, temporarily designated YrZH84, closely linked to SSR markers Xcfa2040-7B and Xbarc32-7B with genetic distances of 1.4 and 4.8 cM, respectively. In a seedling test with 25 PST isolates, the reaction patterns of YrZH84 were different from those of lines carrying Yr2 and Yr6. It was concluded that YrZH84 is probably a new stripe rust resistance gene.     

Characterization and molecular mapping of Yr52 for high-temperature adult-plant resistance to stripe rust in spring wheat germplasm PI 183527

Stripe rust, caused by Puccinia striiformis f. sp. tritici, is one of the most destructive diseases of wheat worldwide. Resistance is the best approach to control the disease. High-temperature adult-plant (HTAP) stripe rust resistance has proven to be race non-specific and durable. However, genes conferring high-levels of HTAP resistance are limited in number and new genes are urgently needed for breeding programs to develop cultivars with durable high-level resistance to stripe rust. Spring wheat germplasm PI 183527 showed a high-level of HTAP resistance against stripe rust in our germplasm evaluations over several years. To elucidate the genetic basis of resistance, we crossed PI 183527 and susceptible wheat line Avocet S. Adult plants of parents, F(1), F(2) and F(2:3) progeny were tested with selected races under the controlled greenhouse conditions and in fields under natural infection. PI 183527 has a single dominant gene conferring HTAP resistance. Resistance gene analog polymorphism (RGAP) and simple sequence repeat (SSR) markers in combination with bulked segregant analysis (BSA) were used to identify markers linked to the resistance gene. A linkage map consisting of 4 RGAP and 7 SSR markers was constructed for the resistance gene using data from 175 F(2) plants and their derived F(2:3) lines. Amplification of nulli-tetrasomic, ditelosomic and deletion lines of Chinese Spring with three RGAP markers mapped the gene to the distal region (0.86-1.0) of chromosome 7BL. The molecular map spanned a genetic distance of 27.3?cM, and the resistance gene was narrowed to a 2.3-cM interval flanked by markers Xbarc182 and Xwgp5258. The polymorphism rates of the flanking markers in 74 wheat lines were 74 and 30?%, respectively; and the two markers in combination could distinguish the alleles at the resistance locus in 82?% of tested genotypes. To determine the genetic relationship between this resistance gene and Yr39, a gene also on 7BL conferring HTAP resistance in Alpowa, a cross was made between PI 183527 and Alpowa. F(2) segregation indicated that the genes were 36.5?±?6.75?cM apart. The gene in PI 183527 was therefore designed as Yr52. This new gene and flanking markers should be useful in developing wheat cultivars with high-level and possible durable resistance to stripe rust.     

Molecular mapping of a recessive gene for resistance to stripe rust in barley

Barley stripe rust, caused by Puccinia striiformis f. sp. hordei, is one of the most important barley (Hordeum vulgare) diseases in the United States. The disease is best controlled using resistant cultivars. Barley genotype Grannenlose Zweizeilige (GZ) has a recessive gene (rpsGZ) that is effective against all races of P. striiformis f. sp. hordei identified so far in the USA. To develop a molecular map for mapping the gene, F₈ recombinant inbred lines (RILs) were developed from the Steptoe X GZ cross through single-seed descent. Seedlings of the parents and RILs were evaluated for resistance to races PSH-14 and PSH-54 of P. striiformis f. sp. hordei under controlled greenhouse conditions. Genomic DNA was extracted from the parents and 182 F₈ RILs and used for linkage analysis. The resistance gene analog polymorphism (RGAP) technique was used to identify molecular markers for rpsGZ. A linkage group for the gene was constructed with 12 RGAP markers, of which two markers co-segregated with the resistance locus, and two markers were closely linked to the locus with a genetic distance of 0.9 and 2.0 cM, respectively. These four markers were present only in the susceptible parent. The closest marker to the resistance allele was 11.7 cM away. Analyses of two sets of barley chromosome addition lines of wheat with the two RGAP markers that were cosegregating with the susceptibility allele showed that rpsGZ and the markers were located on the long arm of barley chromosome 4H. Further, tests with four simple sequence repeat (SSR) markers confirmed the chromosomal location of the rpsGZ gene and also integrated the RGAP markers into the known SSR-based linkage map of barley. The closest SSR marker EBmac0679 had a genetic distance of 7.5 cM with the gene in the integrated linkage map constructed with the 12 RGAP markers and 4 SSR markers. The information on chromosomal location and molecular markers for rpsGZ should be useful for incorporating this gene into commercial cultivars and combining it with other resistance genes for durable resistance.     

Resistance gene-analog polymorphism markers co-segregating with the <Emphasis Type="SmallCaps">YR5</Emphasis> gene for resistance to wheat stripe rust

The Yr5 gene confers resistance to all races of the stripe rust pathogen ( Puccinia striiformis f. sp. tritici) of wheat in the United States. To develop molecular markers for Yr5, a BC(7):F(3) population was developed by backcrossing the Yr5 donor ' Triticum spelta album' (TSA) with the recurrent parent 'Avocet Susceptible' (AVS). Seedlings of the Yr5 near-isogenic lines (AVS/6* Yr5), AVS, TSA, and the BC(7):F(3) lines were tested with North American races of P. striiformis f. sp. tritici under controlled greenhouse conditions. The single gene was confirmed by a 1:2:1 segregation ratio for homozygous-resistant, heterozygous and homozygous-susceptible BC(7):F(3) lines. Genomic DNA was extracted from the parents (the Yr5 near-isogenic line and AVS) and 202 BC(7):F(3) lines. The resistance gene-analog polymorphism (RGAP) technique was used to identify molecular markers. The parents and the homozygous-resistant and homozygous-susceptible BC(7):F(3) bulks were used to identify putative RGAP markers for Yr5. Association of the markers with Yr5 was determined using segregation analysis with DNA from the individual BC(7):F(3) lines. Of 16 RGAP markers confirmed by segregation analysis with 109 BC(7):F(3) lines, and nine of the markers confirmed with an additional 93 BC(7):F(3) lines, three markers co-segregated with the resistance allele and three markers co-segregated with the susceptibility allele at the Yr5 locus. The other four markers were tightly linked to the locus. Analysis of a set of Chinese Spring nulli-tetrasomic lines with three markers that co-segregated with, or were linked to, the susceptibility allele confirmed that the Yr5 locus is on chromosome 2B. Of five RGAP markers that were cloned and sequenced, markers Xwgp-17 and Xwgp-18 that co-segregated with the Yr5 locus were co-dominant and had 98% homology with each other in both DNA and translated amino-acid sequences. The two markers had 97% homology with a resistance gene-like sequence from Aegilops ventricosa and had significant homology with many known plant resistance genes, resistance gene analogs and expressed sequence tags (ESTs) from wheat and other plant species. The markers Xwgp-17 and Xwgp-18 also had significant homology with the NB-ARC domain that is in several genes for plant resistance to diseases, nematode cell death and human apoptotic signaling. These markers should be useful to clone Yr5 and combine Yr5 with other genes for durable and superior resistance for the control of stripe rust.     

Molecular markers for wheat leaf rust resistance gene Lr41

Leaf rust, caused by Puccinia triticina Eriks., is an important foliar disease of common wheat (Triticum aestivum L.) worldwide. Pyramiding several major rust-resistance genes into one adapted cultivar is one strategy for obtaining more durable resistance. Molecular markers linked to these genes are essential tools for gene pyramiding. The rust-resistance gene Lr41 from T. tauschii has been introgressed into chromosome 2D of several wheat cultivars that are currently under commercial production. To discover molecular markers closely linked to Lr41, a set of near-isogenic lines (NILs) of the hard winter wheat cultivar Century were developed through backcrossing. A population of 95 BC₃F_2:6 NILs were evaluated for leaf rust resistance at both seedling and adult plant stages and analyzed with simple sequence repeat (SSR) markers using bulked segregant analysis. Four markers closely linked to Lr41 were identified on chromosome 2DS; the closest marker, Xbarc124, was about 1 cM from Lr41. Physical mapping using Chinese Spring nullitetrasomic and ditelosomic genetic stocks confirmed that markers linked to Lr41 were on chromosome arm 2DS. Marker analysis in a diverse set of wheat germplasm indicated that primers BARC124, GWM210, and GDM35 amplified polymorphic bands between most resistant and susceptible accessions and can be used for marker-assisted selection in breeding programs.     

Characterization and mapping of cryptic alien introgression from <Emphasis Type="Italic">Aegilops geniculata</Emphasis> with new leaf rust and stripe rust resistance genes <Emphasis Type="Italic">Lr57</Emphasis> and <Emphasis Type="Italic">Yr40</Emphasis> in wheat

Leaf rust and stripe rust are important foliar diseases of wheat worldwide. Leaf rust and stripe rust resistant introgression lines were developed by induced homoeologous chromosome pairing between wheat chromosome 5D and 5M^g of Aegilops geniculata (U^gM^g). Characterization of rust resistant BC₂F₅ and BC₃F₆ homozygous progenies using genomic in situ hybridization with Aegilops comosa (M) DNA as probe identified three different types of introgressions; two cytologically visible and one invisible (termed cryptic alien introgression). All three types of introgression lines showed similar and complete resistance to the most prevalent pathotypes of leaf rust and stripe rust in Kansas (USA) and Punjab (India). Diagnostic polymorphisms between the alien segment and recipient parent were identified using physically mapped RFLP probes. Molecular mapping revealed that cryptic alien introgression conferring resistance to leaf rust and stripe rust comprised less than 5% of the 5DS arm and was designated T5DL·5DS-5M^gS(0.95). Genetic mapping with an F₂ population of Wichita × T5DL·5DS-5M^gS(0.95) demonstrated the monogenic and dominant inheritance of resistance to both diseases. Two diagnostic RFLP markers, previously mapped on chromosome arm 5DS, co-segregated with the rust resistance in the F₂ population. The unique map location of the resistant introgression on chromosome T5DL·5DS-5M^gS(0.95) suggested that the leaf rust and stripe rust resistance genes were new and were designated Lr57 and Yr40. This is the first documentation of a successful transfer and characterization of cryptic alien introgression from Ae. geniculata conferring resistance to both leaf rust and stripe rust in wheat.     

Molecular mapping of a stripe rust resistance gene in wheat line C51

Stripe rust, a major disease in areas where cool temperatures prevail, can strongly influence grain yield. To control this disease, breeders have incorporated seedling resistance genes from a variety of sources outside the primary wheat gene pool. The wheat line C51, introduced from the International Center for Agricultural Research in the Dry Areas (ICARDA), Syria, confers resistance to all races of Puccinia striiformis f. sp. tritici (PST) in China. To map the resistant gene(s) against stripe rust in wheat line C51, 212 F ₈ recombinant inbred lines (RILs) derived from the cross X440 × C51 were inoculated with Chinese PST race CYR33 (Chinese yellow rust, CYR) in the greenhouse. The result showed that C51 carried a single dominant gene for resistance (designated YrC51) to CYR33. Simple sequence repeat (SSR) and resistance gene-analogue polymorphism (RGAP) markers that were polymorphic between the parents were used for genotyping the 212 F ₈ RILs. YrC51was closely linked to two SSR loci on chromosome 2BS with genetic distances of 5.1 cM (Xgwm429) and 7.2 cM (Xwmc770), and to three RGAP markers C51R1 (XLRR For / NLRR For), C51R2 (CLRR Rev / Cre3LR-F) and C51R3 (Pto kin4/ NLRR-INV2) with genetic distances of 5.6, 1.6 and 9.2 cM, respectively. These RGAP-linked markers were then converted into STS markers. Among them, one STS marker, C51STS-4, was located at a genetic distance of 1.4 cM to YrC51 and was closely associated with resistance when validated in several populations derived from crosses between C51 and Sichuan cultivars. The results indicated that C51STS-4 can be used for marker assisted selection (MAS) and would facilitate the pyramiding of YrC51 with other genes for stripe rust resistance.