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.     

Development of an STS marker tightly linked to <Emphasis Type="Italic">Yr26</Emphasis> against wheat stripe rust using the resistance gene-analog polymorphism (RGAP) technique

The gene Yr26 confers resistance to all races of Puccinia striiformis f. sp. tritici (PST), the casual pathogen of wheat stripe rust in China. Here, we report development of a molecular marker closely linked to Yr26 using a resistance gene-analog polymorphism (RGAP) technique. A total of 787 F₂ plants and 165 F₃ lines derived from the cross Chuanmai 42/Taichung 29 were used for linkage analysis. Eighteen near-isogenic lines (NILs) and 18 Chinese wheat cultivars and advanced lines with different genes for stripe rust resistance were employed for the validation of STS markers. A total of 1,711 RGAP primer combinations were used to test the parents and resistant and susceptible bulks. Five polymorphic RGAP markers were used for genotyping all F₂ plants. Linkage analysis showed that the five RGAP markers were closely linked to Yr26 with genetic distances ranging from 0.5 to 2.9 cM. These markers were then converted into STS markers, one, CYS-5, of which was located 0.5 cM to Yr26 and was closely associated with the resistance gene when validated over 18 NILs and 18 Chinese wheat cultivars and lines. The results indicated that CYS-5 can be used in marker-assisted selection targeted at pyramiding Yr26 and other genes for stripe rust resistance.     

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.     

<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.     

Molecular mapping of genes for race-specific overall resistance to stripe rust in wheat cultivar Express

‘Express’, a hard red spring wheat cultivar that has been widely grown in the western United States, is used to differentiate races of Puccinia striiformis f. sp. tritici, the causal fungal pathogen of wheat stripe rust. To identify genes conferring race-specific, overall resistance to stripe rust, Express was crossed with ‘Avocet S’. The parents and F₁, F₂, F₃ and F₅ populations were tested with races PST-1, PST-21, PST-43, and PST-45 of P. striiformis f. sp. tritici in the seedling stage under controlled greenhouse conditions. Two dominant genes for resistance to stripe rust were identified, one conferring resistance to PST-1 and PST-21, and the other conferring resistance to all four races. Linkage groups were constructed for the resistance genes using 146 F₅ lines to establish resistance gene analog and chromosome-specific simple sequence repeat marker polymorphisms. The gene for resistance to races PST-1 and PST-21 was mapped on the long arm of chromosome 1B, and that conferring resistance to all four races was mapped on the long arm of chromosome 5B. We temporarily designate the gene on 1BL as YrExp1 and the gene on 5BL as YrExp2. Polymorphism of at least one of the two markers flanking YrExp2 was detected in 91% of the 44 tested wheat genotypes, suggesting that they would be useful in marker-assisted selection for combining the gene with other resistance genes into many other wheat cultivars. Knowledge of these genes will be useful to understand recent virulence changes in the pathogen populations.     

Dr. Heinz rogoll‐70 Jahre

The wheat crop remains vulnerable to all three rust diseases (leaf rust, stem rust and yellow rust) caused by Puccinia spp. according to the prevalence of the pathogen in different wheat-growing areas worldwide. Stripe rust or yellow rust caused by Puccinia striiformis f. sp. tritici is the most significant rust pathogen which prefers cool, moist areas and highlands. The pathogen is recognised as responsible for huge production losses in wheat. Genetic variation in pathogen makes its control difficult. Therefore, resistance against all the races of the pathogen known as durable or race-non-specific resistance is preferred. The present study was carried out to identify durable resistance against stripe rust in selected wheat cultivars from Pakistan through seedling testing, field evaluation at adult stage, morphological marker studies and marker-assisted selection. Results revealed that 4% of the cultivars were resistant at the seedling stage while the rest were susceptible or intermediate. To confirm their field resistance, the same cultivars were evaluated under field conditions at Cereal Crops Research Institute Pirsabak (located in Khyber Pakhtunkhwa, KP) a hot spot of stripe rust in Pakistan. Observations exhibited that at the adult stage 4% of the cultivars were resistant, 70% intermediate or moderately resistant while the others were highly susceptible. Leaf tip necrosis was observed in 30% of the cultivars. Wheat cultivars showing susceptibility at the seedling stage were highly to moderately resistant at adult stage showing durable resistance. For further validation, morphological markers were also observed in cultivars indicating the presence of Yr18/Lr34 gene. Eleven cultivars (C-518, Mexipak, Kohinoor-83, Faisalabad-83, Zardana-93, Shahkar-95, Moomal-2002, Wattan-94, Pasban-90, Kiran-95, and Haider-2000) were identified, having durable or race non-specific resistance against stripe rust. These cultivars can further be utilised in wheat breeding programmes for deploying durable resistance to attain long lasting control against stripe rust.     

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 Yr53, a new gene for stripe rust resistance in durum wheat accession PI 480148 and its transfer to common wheat

Stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), is one of the most damaging diseases of wheat worldwide. It is essential to identify new genes for effective resistance against the disease. Durum wheat PI 480148, originally from Ethiopia, was resistant in all seedling tests with several predominant Pst races in the US under controlled greenhouse conditions and at multiple locations subject to natural infection for several years. To map the resistance gene(s) and to transfer it to common wheat, a cross was made between PI 480148 and susceptible common wheat genotype Avocet S (AvS). Resistant F₃ plants with 42 chromosomes were selected cytologically and by testing with Pst race PST-100. A total of 157 F₄ plants from a single F₃ plant with 2n = 42 tested with PST-100 segregated in a 3 resistant: 1 susceptible ratio, indicating that a single dominant gene from PI 480148 conferred resistance. Using the F_3:4 population and the resistance gene-analog polymorphism (RGAP) and simple sequence repeat (SSR) markers, the gene was mapped to the long arm of chromosome 2B. SSR marker Xwmc441 and RGAP marker XLRRrev/NLRRrev ₃₅₀ flanked the resistance gene by 5.6 and 2.7 cM, respectively. The effective resistance of the gene to an Australian Pst isolate virulent to Yr5, which is also located on 2BL and confers resistance to all US Pst races, together with an allelism test of the two genes, indicated that the gene from PI 480148 is different from Yr5 and should be a new and useful gene for resistance to stripe rust. Resistant common wheat lines with plant types similar to AvS were selected for use in breeding programs.     

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.     

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.     

Reaction of some wheat cultivars and breeding lines to Puccinia striiformis f. sp. tritici hot races in Iran

Many physiological races of Puccinia striiformis f. sp. tritici which cause stripe rust in wheat can be determined in different parts of the world. The emergence of new races with different pathogenicity which happens very quickly breaks cultivars resistant and cause disease. Therefore, breeding cultivar for resistance to different pathogenic races should be continued. In this research, pathogenicity of two isolates collected from two regions of Iran were determined by using wheat yellow rust differential lines, which indicated race 70E50A+ and 6E18A+ The responses of 30 wheat genotypes were separately evaluated in the forms of randomized complete block design with three replicates in the seedling stage under greenhouse condition. The components of resistance including latent period and infection type were recorded. Results indicated genotypes were evaluated in terms of both traits and were significant at 1% level. Also, the results from pathogenicity study indicated of effective gene/s included Yr1, Yr2+, Yr3, Yr4, Yr5, Yr10, Yr15, Yr24, Yr26, YrSP, YrND, YrSD and YrSU. From the genotypes studied in the greenhouse condition, 39% of the genotypes showed complete resistance to both races. Probably, resistance genes, Yr32 and YrCV, or the other unknown genes which are types of seedling resistance are either alone or in combination of one another cause strength in resistant genotypes.     

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.     

Powdery mildew resistance and Lr34/Yr18 genes for durable resistance to leaf and stripe rust cosegregate at a locus on the short arm of chromosome 7D of wheat

The incorporation of effective and durable disease resistance is an important breeding objective for wheat improvement. The leaf rust resistance gene Lr34 and stripe rust resistance gene Yr18 are effective at the adult plant stage and have provided moderate levels of durable resistance to leaf rust caused by Puccinia triticina Eriks. and to stripe rust caused by Puccinia striiformis Westend. f. sp. tritici. These genes have not been separated by recombination and map to chromosome 7DS in wheat. In a population of 110 F₇ lines derived from a Thatcher × Thatcher isogenic line with Lr34/Yr18, field resistance to leaf rust conferred by Lr34 and to stripe rust resistance conferred by Yr18 cosegregated with adult plant resistance to powdery mildew caused by Blumeria graminis (DC) EO Speer f. sp. tritici. Lr34 and Yr18 were previously shown to be associated with enhanced stem rust resistance and tolerance to barley yellow dwarf virus infection. This chromosomal region in wheat has now been linked with resistance to five different pathogens. The Lr34/Yr18 phenotypes and associated powdery mildew resistance were mapped to a single locus flanked by microsatellite loci Xgwm1220 and Xgwm295 on chromosome 7DS.     

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%.     

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.     

Identification and characterization of stripe rust resistance gene Yr34 in common wheat

An uncharacterized source of seedling resistance to Puccinia striiformis f.sp. tritici was identified in an advanced wheat breeding line WAWHT2046. Genetic analysis based on a WAWHT2046/Carnamah-derived double haploid (DH) population demonstrated monogenic inheritance of seedling stripe rust resistance in WAWHT2046. The gene controlling stripe rust resistance in line WAWHT2046 was tentatively designated YrWA. The chromosome 5AL located awn inhibitor gene B1, possessed by WAWHT2046, also showed monogenic inheritance when the DH population was scored for the presence and absence of awns. Joint segregation analysis at the B1 and YrWA loci indicated genetic linkage between the two loci. A recombination value of 12.2 cM was computed using Mapmanager. This association located YrWA in the chromosome arm 5AL. Molecular mapping using microsatellite markers placed YrWA distal to B1. All molecular markers mapped proximal to the awn inhibitor locus B1. As no other stripe rust resistance gene is reported to be located in the chromosome arm 5AL, YrWA was permanently designated as Yr34. Yr34 produced an intermediate (23C) seedling infection type and expressed very low stripe rust response (10R-MR) on adult plants in the field, similar to the resistance gene Yr17. In addition to Yr34, this mapping population segregated for three genetically independent adult plant stripe rust resistance genes. The detection of DH lines with completely susceptible response, higher than that shown by the Yr34-lacking parent Carnamah, suggested that both parents contributed adult plant resistance. The use of WAWHT2046 as a parent in breeding programs would also contribute APR in addition to Yr34.     

Development and characterization of novel microsatellite markers in Puccinia striiformis f.sp. tritici and their transferability in Puccinia species

Puccinia striiformis f.sp. tritici (Pst) and P. striiformis f.sp. hordei (Psh) causing stripe rust disease in wheat and barley, respectively, are two devastating phytopathogens. Microsatellite/simple sequence repeat (SSR) markers are increasingly being utilized for analysis of genetic diversity, diagnosis, population structure and possible migratory routes of plant pathogens. In the current study, novel polymorphic SSR markers were designed for Pst using the genomic sequences of PST-78 isolate. A total of 1,191 SSR motifs, comprising 30% each of di- and tri-nucleotide type of repeats, 17% of penta-nucleotide, 15% of tetra-nucleotide and 8% of hexa-nucleotide repeats, were detected through in silico scanning of PST-78 genomic sequences. Polymorphism was detected by nine of the 50 designed SSRs (PsSSRs) in seven stripe rust pathotypes of wheat and barley. The mean number of alleles per SSR locus, mean polymorphism information content (PIC), mean heterozygosity, mean major allele frequency (MAF) and mean gene diversity were 2.33, 0.34, 0.33, 0.71 and 0.40, respectively. The dendrogram analysis suggested that newly developed PsSSR markers could distinguish stripe rust pathotypes based on their virulence phenotype. Further, the cross-genera and cross-species amplification test of these markers in 14 different rust pathotypes revealed that 9 PsSSRs are capable of amplification in Pst species infecting wild grass, followed by 6 PsSSRs in Pt, 3 PsSSRs in Pgt, 1 PsSSRs in Puccinia species on barberry and Melampsora lini. Thus, the transferability of PsSSRs to other species reduced with increasing genetic distance of target species. These newly designed SSR markers expand the available Pst SSR marker resources and allow better genetic studies.     

Backcross-breeding and doubled-haploid facilitated introgression of stripe rust resistance in bread wheat

Stripe rust caused by the fungus Puccinia striiformis f. sp. tritici (Pst) may decrease wheat yield significantly in severe outbreaks. The most cost-effective and environmentally friendly approach to reduce yield losses due to rust diseases is deployment of effective resistant genes in wheat cultivars. The causal agents evolve and may break existing resistant sources as well. Therefore, long-term conventional breeding strategies and the ongoing evolution of pathogen populations in the region would put the success of breeding programmes at risk so that there is always a need for speeding up the process of germplasm enhancement through production of doubled-haploid breeding materials. In this study, we aimed at introgression of stripe rust resistance trait from three genotypes (Flanders, Martonvasar-17 (MV17) and Bersee) into a widely adapted cultivar “Ghods”. Positively selected F2BC² progenies of three backcrossing schemas, i.e. (i) Flanders/3^*Ghods; (ii) Ghods^*3/MV17; and (iii) Hybride-de-bersee/3^*Ghods, were used to produce three small-size doubled-haploid populations via wheat × Maize pollination methodology. The doubled-haploid populations were examined against two predominantly isolates of P. striiformis f. sp. tritici (Pst) i.e. 6E134A⁺ and 6E2A⁺Yr27⁺ and the screening revealed that 44 and 52 of the progenies are resistant to the above-mentioned isolates, respectively. Field data have shown that the stripe rust resistance doubled-haploid germplasm are comparable to local check cultivars in yield and earliness.     

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.