Intercellular chitinase and peroxidase activities associated with resistance conferred by geneLr35to leaf rust of wheat

The effect of leaf rust (Puccinia triticina) infection on intercellular chitinase (EC 3.2.1.14) and peroxidase (EC 1.11.1.7) activities was studied in resistant [RL 6082 (Thatcher/Lr35)] and susceptible (Thatcher) near isogenic wheat (Triticum aestivum L.) lines at seedling, stem elongation and flag leaf stages of plant growth. The levels of activity of these enzymes were low during the seedling and stem elongation stages. Resistant plants at the flag leaf stage, during which the Lr35 resistance gene was maximally expressed, exhibited high constitutive levels of chitinase and peroxidase activities, in contrast to the lower constitutive levels of susceptible plants. The results suggest that chitinase and peroxidase, constitutively present in the intercellular spaces of Thatcher/Lr35 wheat leaves, may play a role in Lr35 mediated resistance to leaf rust.     

An introgression on wheat chromosome 4DL in RL6077 (Thatcher*6/PI 250413) confers adult plant resistance to stripe rust and leaf rust (Lr67)

Adult plant resistance (APR) to leaf rust and stripe rust derived from the wheat (Triticum aestivum L.) line PI250413 was previously identified in RL6077 (=Thatcher*6/PI250413). The leaf rust resistance gene in RL6077 is phenotypically similar to Lr34 which is located on chromosome 7D. It was previously hypothesized that the gene in RL6077 could be Lr34 translocated to another chromosome. Hybrids between RL6077 and Thatcher and between RL6077 and 7DS and 7DL ditelocentric stocks were examined for first meiotic metaphase pairing. RL6077 formed chain quadrivalents and trivalents relative to Thatcher and Chinese Spring; however both 7D telocentrics paired only as heteromorphic bivalents and never with the multivalents. Thus, chromosome 7D is not involved in any translocation carried by RL6077. A genome-wide scan of SSR markers detected an introgression from chromosome 4D of PI250413 transferred to RL6077 through five cycles of backcrossing to Thatcher. Haplotype analysis of lines from crosses of Thatcher × RL6077 and RL6058 (Thatcher*6/PI58548) × RL6077 showed highly significant associations between introgressed markers (including SSR marker cfd71) and leaf rust resistance. In a separate RL6077-derived population, APR to stripe rust was also tightly linked with cfd71 on chromosome 4DL. An allele survey of linked SSR markers cfd71 and cfd23 on a set of 247 wheat lines from diverse origins indicated that these markers can be used to select for the donor segment in most wheat backgrounds. Comparison of RL6077 with Thatcher in field trials showed no effect of the APR gene on important agronomic or quality traits. Since no other known Lr genes exist on chromosome 4DL, the APR gene in RL6077 has been assigned the name Lr67.     

Undescribed wheat gene for partial leaf rust and stripe rust resistance from Thatcher derivatives RL6058 and 90RN249 carryingLr34

Inheritance of partial leaf rust and stripe rust resistance of a Thatcher wheat 90RN2491, earlier reported to carry two doses of the gene pairLr34-Yr18 and the reference line RL6058 (6*Thatcher/PI58548) for theLr34-Yr18 gene pair was studied against predominant and highly virulent Indian races. Thatcher derivatives 90RN2491 and RL6058 were intercrossed as well as crossed with the leaf rust and stripe rust susceptible Indian cultivar WL711. The F₁, F₂ and F₃ generations from these crosses were assessed for rust severity against leaf rust race 77-5 and stripe rust race 46S119. The F₂ and F₃ generations from the crosses of RL6058 and 90RN2491 with WL711, segregated 15 resistant : 1 susceptible (F₂) and 7 homozygous resistant : 8 segregating : 1 homozygous susceptible (F₃) ratios, respectively, both for leaf rust and stripe rust severity. Therefore, partial resistance against each of the leaf rust and stripe rust races in both RL6058 and 90RN2491 is ascribed to two independently inherited dominant genes. One of the two genes for leaf rust and stripe rust resistance in 90RN2491 and RL6058 isLr34 and the linked geneYr18, respectively. The second leaf rust resistance gene in both the Thatcher lines segregated independently of stripe rust resistance. Therefore, it is notLr34 and it remains unidentified.     

Identification of molecular markers for the detection of the yellow rust resistance gene Yr17 in wheat

The Yr17 gene, which is present in many European wheat cultivars, displays yellow rust resistance at the seedling stage. The gene introduced into chromosome 2A from Aegilops ventricosa was previously found to be closely linked (0.5 cM) to leaf and stem rust resistance genes Lr37 and Sr38, respectively. The objective of this study was to identify molecular markers linked to the Yr17 gene. We screened with RAPD primers, for polymorphism, the DNAs of cv. Thatcher and the leaf rust-resistant near-isogenic line (NIL) RL 6081 of cv. Thatcher carrying the Lr37 gene. Using a F2 progeny of the cross between VPM1 (resistant) and Thésée (susceptible), the RAPD marker OP-Y15580 was found to be closely linked to the Yr17 gene. We converted the OP- Y15580 RAPD marker into a sequence characterized amplified region (SCAR). This SCAR marker (SC-Y15) was linked at 0.8 ± 0.7 cM to the Yr17 resistance gene. We tested the SC-Y15 marker over a survey of 37 wheat cultivars in order to verify its consistency in different genetic backgrounds and to explain the resistance of some cultivars against yellow rust. Moreover, we showed that the Xpsr150-2Mv locus marker of Lr gene described by Bonhomme et al. [6] which possesses A. ventricosa introgression on the 2A chromosome was also closely linked to the Yr17 gene. Both the SCAR SC-Y15 and Xpsr150-2Mv markers should be used in breeding programmes in order to detect the cluster of the three genes Yr17, Lr37 and Sr38 in cross progenies. This revised version was published online in June 2006 with corrections to the Cover Date.     

Intercellular proteins and β-1,3-glucanase activity associated with leaf rust resistance in wheat

To investigate biochemical aspects of resistance conferred by the Lr35 gene for adult-plant resistance in wheat ( Triticum aestivum L.) to leaf rust, pathogen development was related to intercellular protein composition and β -1,3-glucanase (EC 3.2.1.39) activities at three growth stages in infected and uninfected resistant (RL6082 [Thatcher/ Lr35 ]) and susceptible (Thatcher) plants. Leaf rust symptoms produced by pathotype UVPrt9 of Puccinia recondita f. sp. tritici showed that resistance conferred by Lr35 was most effective at the flag leaf stage. Furthermore, fluorescence microscopy indicated that resistance was strongly associated with hypersensitive cell death of invaded tissue. According to polypeptide profiles, intercellular proteins with molecular masses of 35, 33, 31 and 26 kDa were constitutively present at higher levels in resistant than in susceptible plants at the flag leaf stage. Four intercellular proteins (35, 33, 32 and 31 kDa) serologically related to β -1,3-glucanase were present in resistant and susceptible genotypes during all stages of plant growth. Resistance was associated with high constitutive levels of β -1,3-glucanase activity. Susceptibility on the other hand was associated with low constitutive levels of β -1,3-glucanase, while high levels were induced by infection during more advanced stages of colonization. Our results suggest that β -1,3-glucanase is involved in the defense response controlled by the Lr35 gene.     

Genetic and physical characterization of theLR1 leaf rust resistance locus in wheat (Triticum aestivum L.)

The objective of this study was to characterize the leaf rust resistance locusLr1 in wheat. Restriction fragment length polymorphism (RELP) analysis was performed on the resistant lineLr1/6^*Thatcher and the susceptible varieties Thatcher and Frisal, as well as on the segregating F₂ populations. Seventeen out of 37 RFLP probes mapping to group 5 chromosomes showed polymorphism betweenLr1/6^*Thatcher and Frisal, whereas 11 probes were polymorphic between the near-isogenic lines (NILs)Lr1/6^*Thatcher and Thatcher. Three of these probes were linked to the resistance gene in the segregating F₂ populations. One probe (pTAG621) showed very tight linkage toLr1 and mapped to a single-copy region on chromosome 5D. The map location of pTAG621 at the end of the long arm of chromosome 5D was confirmed by the absence of the band in the nulli-tetrasomic line N5DT5B of Chinese Spring and a set of deletion lines of Chinese Spring lacking the distal part of 5DL. Twenty-seven breeding lines containing theLr1 resistance gene in different genetic backgrounds showed the same band asLr1/6^*Thatcher when hybridized with pTAG621. The RFLP marker was converted to a sequence-tagged-site marker using polymerase chain reaction (PCR) amplification. Sequencing of the specific fragment amplified from both NILs revealed point mutations as well as small insertion/deletion events. These were used to design primers that allowed amplification of a specific product only from the resistant lineLr1/6^*Thatcher. This STS, specific for theLr1 resistance gene, will allow efficient selection for the disease resistance gene in wheat breeding programmes. In addition, the identification of a D-genome-specific probe tightly linked toLr1 should ultimately provide the basis for positional cloning of the gene.     

Locating the broad-spectrum wheat leaf rust resistance gene Lr52 (LrW) to chromosome 5B by a new cytogenetic method

This study was conducted to genetically map a potentially new wheat leaf rust resistance gene (LrW) using a novel genetic method and to test its effectiveness against current races of leaf rust (Puccinia triticina Eriks.) in Canada. Undoubled haploids of a near-isogenic line of Thatcher carrying the resistance gene (RL6107) were pollinated with a contrasting susceptible cultivar to generate an array of hybrids with random deficiencies arising from irregular meiosis of the haploid. Genetic analysis of the deficiencies in such populations can be used to locate qualitative traits by which the two parents differ through a process that we have called haploid deficiency mapping. In the present case, 5/417 hybrids were both susceptible to leaf rust (i.e. lacked the resistance gene) and also lacked several polymorphic microsatellite alleles from RL6107 that are specific to chromosome 5B. This correlated failed transmission of the resistance gene and deficiency for chromosome 5B. Analysis of an F₂ population showed that the factor conditioning resistance was located on the short arm of 5B, 16.5 cM distal to the locus of the microsatellite Xgwm443. Since no other leaf rust resistance genes have been mapped to this region, LrW was re-designated Lr52. RL6107 was tested with 29 isolates of P. triticina, encompassing a diversity of virulence found in North America, with none showing virulence. The effectiveness and novelty of Lr52 make it a promising source of resistance for North American wheat cultivars.     

Identification of microsatellite markers linked to leaf rust resistance gene <Emphasis Type="Italic">Lr25</Emphasis> in wheat

The leaf rust resistance gene Lr25, transferred from Secale cereale L. into wheat and located on chromosome 4B, imparts resistance to all pathotypes of leaf rust in South-East Asia. In an F₂-derived F₃ population, created by crossing TcLr25 that carries the gene Lr25 for leaf rust resistance with leaf rust-susceptible parent Agra Local, three microsatellite markers located on the long arm of chromosome 4B were found to be linked to the Lr25 locus. The donor parent TcLr25 is a near-isogenic line derived from the variety Thatcher. The most virulent pathotype of leaf rust in the South-East Asian region, designated 77–5 (121R63-1), was used for challenging the population under artificially controlled conditions. The marker Xgwm251 behaved as a co-dominant marker placed 3.8 cM away from the Lr25 locus on 4BL. Two null allele markers, Xgwm538 and Xgwm6, in the same linkage group were located at a distance of 3.8 cM and 16.2 cM from the Lr25 locus, respectively. The genetic sequence of Xgwm251, Lr25, Xgwm538, and Xgwm6 covered a total length of 20 cM on 4BL. The markers were validated for their specificity to Lr25 resistance in a set of 43 wheat genetic stocks representing 43 other Lr genes.     

Occurrence of wheat leaf rust (Puccinia triticina) races and virulence changes in Slovakia in 1994–2004

In 1995–2004 we investigated leaf rust virulence in Slovakia on Thatcher near isogenic lines (NILs) with genes Lr1, Lr2a, Lr2b, Lr2c, Lr3a, Lr9, Lr10, Lr11, Lr15, Lr17, Lr19, Lr21, Lr23, Lr24, Lr26 and Lr28. According to reaction of leaf rust isolates resistance genes Lr9 and Lr19 were completely effective to all examined pathotypes in all years. The resistance genes Lr24 and Lr28 were also completely effective to all examined pathotypes till the year 2001. In the year 2001 we detected 20% and 10% virulent isolates on NILs Lr24 and Lr28, respectively. According to the reaction of investigated isolates from the territory of Slovakia on NILs, resistance genes Lr2c, Lr3a, Lr11, Lr17, Lr21, Lr23 and Lr26 were mostly ineffective. During the 1994–2004 period we detected 16 races of leaf rust (races 2, 2SaBa, 6, 6SaBa, 12, 12SaBa, 14, 14SaBa, 57, 57SaBa, 61, 61SaBa, 62SaBa, 77, 77SaBa, 77/57SaBa). The most frequently determined races were 61SaBa and 77SaBa, which occurred in all years. Among frequently determined races we can assign race 12SaBa as well. According to the field tests in 2001–2004 good resistance to leaf rust was displayed by the cvs Arida (Lr13, Lru), Eva (Lr3, Lru) and Solara (Lru).     

Application of STS markers for leaf rust resistance genes in near-isogenic lines of spring wheat cv. Thatcher

Sequence tagged site (STS) markers for eight resistance genes against Puccinia recondita f. sp. tritici were used to screen a set of near-isogenic lines of wheat cv. Thatcher containing in total 40 different Lr genes and their alleles. Polymerase chain reaction (PCR) analysis was carried out by using STS, SCAR and CAPS primers specific for the leaf rust resistance genes Lr1, Lr9, Lr10, Lr19, Lr24, Lr28, Lr37 and Lr47. The STS, CAPS and SCAR markers linked to resistance genes Lr9, Lr10, Lr19, Lr24, Lr37 and Lr47 were found to be reliable in diverse genetic backgrounds. The amplification product of the Lr1 gene marker was detected in the susceptible cv. Thatcher and in all of the near-isogenic lines examined except Lr2a, Lr2b, Lr2c and Lr19. The sequence analysis of PCR products amplified in lines Lr1, Lr10, Lr28 and in cv. Thatcher indicated that the near-isogenic lines and cv. Thatcher contained in the targeted chromosome region an allele that differed from the original alleles corresponding to Lr1/6*Thatcher (TLR621) and susceptible Thatcher (TH621). The amplification product specific to the STS marker of the Lr1 gene was amplified in almost all Thatcher near-isogenic lines and in cv. Thatcher because their alleles possessed primer sequences identical to the original allele TLR621. The marker for the Lr28 resistance gene was identified in line Lr28, carrying gene Lr28, and in 21 other near-isogenic lines. The sequencing of PCR products specific to Lr28 and generated in lines Lr1, Lr10 and Lr28 indicated that the lines Lr1, Lr10 and Lr28 are heterozygous in this region.     

Genetic and physical characterization of theLR1 leaf rust resistance locus in wheat (Triticum aestivum L.)

The objective of this study was to characterize the leaf rust resistance locusLr1 in wheat. Restriction fragment length polymorphism (RELP) analysis was performed on the resistant lineLr1/6^*Thatcher and the susceptible varieties Thatcher and Frisal, as well as on the segregating F₂ populations. Seventeen out of 37 RFLP probes mapping to group 5 chromosomes showed polymorphism betweenLr1/6^*Thatcher and Frisal, whereas 11 probes were polymorphic between the near-isogenic lines (NILs)Lr1/6^*Thatcher and Thatcher. Three of these probes were linked to the resistance gene in the segregating F₂ populations. One probe (pTAG621) showed very tight linkage toLr1 and mapped to a single-copy region on chromosome 5D. The map location of pTAG621 at the end of the long arm of chromosome 5D was confirmed by the absence of the band in the nulli-tetrasomic line N5DT5B of Chinese Spring and a set of deletion lines of Chinese Spring lacking the distal part of 5DL. Twenty-seven breeding lines containing theLr1 resistance gene in different genetic backgrounds showed the same band asLr1/6^*Thatcher when hybridized with pTAG621. The RFLP marker was converted to a sequence-tagged-site marker using polymerase chain reaction (PCR) amplification. Sequencing of the specific fragment amplified from both NILs revealed point mutations as well as small insertion/deletion events. These were used to design primers that allowed amplification of a specific product only from the resistant lineLr1/6^*Thatcher. This STS, specific for theLr1 resistance gene, will allow efficient selection for the disease resistance gene in wheat breeding programmes. In addition, the identification of a D-genome-specific probe tightly linked toLr1 should ultimately provide the basis for positional cloning of the gene.     

Phenotypic and genetic diversity of leaf rust resistance in wheat wild relatives

Puccinia triticina (Pt), the causal agent of leaf rust evolves through forming new pathotypes that adversely affect the growth and yield of wheat cultivars. Therefore, continued production of resistant varieties through exploring novel sources of resistance in wild relatives which are abundantly found in Iran and the neighbouring regions is a major task in wheat breeding programs. The aim of the present study was to explore 60 wild wheat genotypes selected from the species Triticum monococcum, Aegilops tauschii, Ae. neglecta, Ae. cylindrica, Ae. triuncialis, Ae. umbellulata, Ae. speltoides, Ae. columnaris, Ae. crassa and Ae. ventricosa for resistance to leaf rust. The cultivar ‘Boolani’ and Thatcher near-isogenic lines were used as controls. Two-week-old seedlings were inoculated using 10 Pt pathotypes, and the infection types were recorded. The genotypes were also analysed for polymorphism using six sequence-tagged sites (STS) and sequence characterized amplified region (SCAR) markers. Forty-eight genotypes produced high infection types (3⁺) for two pathotypes, but the remaining genotypes produced low infection types of ‘0; ^=’ to ‘1⁺CN’ to all pathotypes. The latter included three accessions of Ae. tauschii, two accessions of each Ae. umbellulata, Ae. columnaris and Triticum monococcum, and one accession from each Ae. triuncialis, Ae. ventricosa and Ae. neglecta. Analysis for STS and SCAR markers suggested several genotypes could carry the genes Lr9, Lr10, Lr19, Lr24, Lr26 and Lr37 or their potential orthologs in addition to unknown resistance genes. In conclusion, the identified resistant genotypes could be further characterized and used in wheat breeding programs for leaf rust resistance.     

Phenotypic and molecular analyses of leaf rust resistance in some Iranian wheat genotypes

Host resistance is the most sustainable method of controlling leaf rust which can be achieved through exploring resistance genes by gene postulation and/or molecular markers. The experiment was conducted to postulate leaf rust resistance genes in 20 Iranian wheat cultivars using 10 Puccinia triticina pathotypes. Six sequence-tagged site and sequence-characterised amplified region markers were also used to detect the genes Lr9, Lr10, Lr19, Lr24, Lr26 and Lr37. The genes Lr3a, Lr3Ka, Lr10, Lr15, Lr19, Lr26, Lr28, Lr30 and Lr27 + Lr31 genes were postulated to be present either singly or in combination. The cultivars Toos and Dabira were found to have no effective seedling resistance gene(s); The former was shown to carry none of the genes, while the latter carried Lr10, Lr24 and Lr37 based on molecular markers. It was not possible to postulate resistance genes in Sirvan, Backcross Roshan, Zagross and Chamran cultivars. However, molecular association indicated the presence of Lr19, Lr10 and Lr24 in Sirvan, Backcross Roshan, and Chamran, respectively while none in Zagross.     

Functional variability of the Lr34 durable resistance gene in transgenic wheat

Breeding for durable disease resistance is challenging, yet essential to improve crops for sustainable agriculture. The wheat Lr34 gene is one of the few cloned, durable resistance genes in plants. It encodes an ATP binding cassette transporter and has been a source of resistance against biotrophic pathogens, such as leaf rust (Puccinina triticina), for over 100 years. As endogenous Lr34 confers quantitative resistance, we wanted to determine the effects of transgenic Lr34 with specific reference to how expression levels affect resistance. Transgenic Lr34 wheat lines were made in two different, susceptible genetic backgrounds. We found that the introduction of the Lr34 resistance allele was sufficient to provide comparable levels of leaf rust resistance as the endogenous Lr34 gene. As with the endogenous gene, we observed resistance in seedlings after cold treatment and in flag leaves of adult plants, as well as Lr34‐associated leaf tip necrosis. The transgene‐based Lr34 resistance did not involve a hypersensitive response, altered callose deposition or up‐regulation of PR genes. Higher expression levels compared to endogenous Lr34 were observed in the transgenic lines both at seedling as well as adult stage and some improvement of resistance was seen in the flag leaf. Interestingly, in one genetic background the transgenic Lr34‐based resistance resulted in improved seedling resistance without cold treatment. These data indicate that functional variability in Lr34‐based resistance can be created using a transgenic approach.     

Virulence Frequences of Puccinia triticina in Germany and the European Regions of the Russian Federation

From 2001 to 2003, leaf rust was collected in different regions of Germany and the Russian Federation to generate single spore isolates and to study the structure of the pathogen populations by analyses of virulence. The virulence of isolates was tested with 38 near‐isogenic lines each carrying a different resistance gene. The analyses of variance revealed significant effects for the frequency of virulent isolates, the regions and most interactions with years and regions, but no significance was found for the effects of years. In Germany, an increase of virulence frequencies was detected for Lr1 and Lr2a while a decrease was found for Lr3a, Lr3bg and Lr3ka. Such clear trends did not occur in Russia which may be due to the great agroclimatic differences between regions. The variance of the frequency of virulent isolates was used to estimate adequate sample sizes for the analysis of regional populations of leaf rust. This procedure resulted in more reliable information about the dynamic processes within the pathogen populations. In 2002 and 2003, all pathotypes in Germany had a combined virulence to Lr1, Lr2a, Lr2b, Lr15, Lr17 and Lr20 supplemented by a few other genes. The complexity of virulence was lower in the most frequent pathotypes. In Russia virulence to the alleles at locus Lr3 was very common. Using detached leaf segments in Germany and Russia it turned out that the most virulent pathotypes carry 34 and 32 virulence genes, respectively. Virulence to Lr9, Lr19, Lr24 and Lr38 was rare or even absent. The use of major genes, not overcome by corresponding virulent pathotypes, may contribute to more durable types of resistance in case they are combined with genes having different effects, e.g. adult plant resistance.     

An interchromosomal reciprocal translocation in wheat involving leaf rust resistance gene Lr34.

'Thatcher' backcross lines RL6058 and RL6077 have adult-plant leaf rust resistance and were believed to have Lr34. However, genetic analysis revealed that the genes in the two lines were independent of each other. Previous work demonstrated that Lr34 is located on chromosome 7D. The leaf rust resistance gene in RL6058 must be on chromosome 7DS because no recombinants were observed between it and gene Lr29, known to be on chromosome 7DS. It was also linked with Rc3 (30.25 +/- 2.88%), a gene for purple coleoptile on chromosome 7DS. It was independent of Lr19 and NS1 (nonsuppressor mutant), which are located on 7DL. The leaf rust resistance gene in RL6077 was independent of genes Lr19 and Lr29. The presence of quadrivalents in pollen mother cells of the RL6058/RL6077 hybrid indicates that the Lr34 gene in RL6077 may have been translocated onto another chromosome. Lr34 from RL6058 and RL6077 may have been combined in four F3 lines derived from their intercross.     

Mapping and characterization of the new adult plant leaf rust resistance gene <Emphasis Type="Italic">Lr77</Emphasis> derived from Santa Fe winter wheat

Key message

A new gene for adult plant leaf rust resistance in wheat was mapped to chromosome 3BL. This gene was designated as Lr77.

Abstract

‘Santa Fe’ is a hard red winter cultivar that has had long-lasting resistance to the leaf rust fungus, Puccinia triticina. The objective of this study was to determine the chromosome location of the adult plant leaf rust resistance in Santa Fe wheat. A partial backcross line of ‘Thatcher’ (Tc) wheat with adult plant leaf rust resistance derived from Santa Fe was crossed with Thatcher to develop a Thatcher//Tc*2/Santa Fe F₆ recombinant inbred line (RIL) population. The RIL population and parental lines were evaluated for segregation of leaf rust resistance in three field plot tests and in an adult plant greenhouse test. A genetic map of the RIL population was constructed using 90,000 single-nucleotide polymorphism (SNP) markers with the Illumina Infinium iSelect 90K wheat bead array. A significant quantitative trait locus for reduction of leaf rust severity in all four tests was found on chromosome 3BL that segregated as a single adult plant resistance gene. The RILs with the allele from the resistant parent for SNP marker IWB10344 had lower leaf rust severity and a moderately resistant to moderately susceptible response compared to the susceptible RILs and Thatcher. The gene derived from Santa Fe on chromosome 3BL was designated as Lr77. Kompetitive allele-specific polymerase chain reaction assay markers linked to Lr77 on 3BL should be useful for selection of wheat germplasm with this 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.     

Identification and validation of molecular markers linked to the leaf rust resistance gene Lr19 in wheat

A leaf rust resistance gene Lr19 on the chromosome 7DL of wheat derived from Agropyron elongatum was tagged with random amplified polymorphic DNA (RAPD) and microsatellite markers. The F₂ population of 340 plants derived from a cross between the leaf rust resistant near-isogenic line (NIL) of Thatcher (Tc + Lr19) and leaf rust susceptible line Agra Local that segregated for dominant monogenic leaf rust resistance was utilized for generating the mapping population. The molecular markers were mapped in the F₂ derived F₃ homozygous population of 140 seedlings. Sixteen RAPD markers were identified as linked to the alien gene Lr19 among which eight were in a coupling phase linkage. Twelve RAPD markers co-segregated with Lr19 locus. Nine microsatellite markers located on the long arm of chromosome 7D were also mapped as linked to the gene Lr19, including 7 markers which co-segregated with Lr19 locus, thus generating a saturated region carrying 25 molecular markers linked to the gene Lr19 within 10.2 ± 0.062 cM on either side of the locus. Two RAPD markers S265₅₁₂ and S253₇₃₇ which flanked the locus Lr19 were converted to sequence characterized amplified region markers SCS265₅₁₂ and SCS253₇₃₆, respectively. The marker SCS265₅₁₂ was linked with Lr19 in a coupling phase and the marker SCS253₇₃₆ was linked in a repulsion phase, which when used together mimicked one co-dominant marker capable of distinguishing the heterozygous resistant seedlings from the homozygous resistant. The molecular markers were validated on NILs mostly in Thatcher background isogenic for 44 different Lr genes belonging to both native and alien origin. The validation for polymorphism in common leaf rust susceptible cultivars also confirmed the utility of these tightly linked markers to the gene Lr19 in marker-assisted selection.     

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.