Leaf Rust Resistance Gene Lr19 Research Articles

Genetic mapping of the wheat leaf rust resistance gene Lr19 and development of translocation lines to break its linkage with yellow pigment.

The leaf rust resistance gene Lr19, which is present on the long arm of chromosome 7E1 in Thinopyrum ponticum, was mapped within a 0.3-cM genetic interval, and translocation lines were developed to break its linkage with yellow pigmentation The leaf rust resistance locus Lr19, which was transferred to wheat (Triticum aestivum) from its relative Thinopyrum ponticum in 1966, still confers broad resistance to most known races of the leaf rust pathogen Puccinia triticina (Pt) worldwide. However, this gene has not previously been fine-mapped, and its tight linkage with a gene causing yellow pigmentation has limited its application in bread wheat breeding. In this study, we genetically mapped Lr19 using a bi-parental population from a cross of two wheat-Th. ponticum substitution lines, the Lr19-carrying line 7E1(7D) and the leaf rust-susceptible line 7E2(7D). Genetic analysis of the F2 population and the F2:3 families showed that Lr19 was a single dominant gene. Genetic markers allowed the gene to be mapped within a 0.3-cM interval on the long arm of Th. ponticum chromosome 7E1, flanked by markers XsdauK3734 and XsdauK2839. To reduce the size of the Th. ponticum chromosome segment carrying Lr19, the Chinese Spring Ph1b mutant was employed to promote recombination between the homoeologous chromosomes of the wheat chromosome 7D and the Th. ponticum chromosome 7E1. Two translocation lines with short Th. ponticum chromosome fragments carrying Lr19 were identified using the genetic markers closely linked to Lr19. Both translocation lines were resistant to 16 Pt races collected throughout China. Importantly, the linkage between Lr19 and yellow pigment content was broken in one of the lines. Thus, the Lr19 linked markers and translocation lines developed in this study are valuable resources in marker-assisted selection as part of common wheat breeding programs.

Multiplex PCR assay for the simultaneous identification of race specific and non-specific leaf resistance genes in wheat (Triticum aestivum L.)

Race-nonspecific resistance is a key to sustainable management of pathogens in bread wheat (Triticum aestivum L.) breeding. It is more durable compared to race-specific immunity, conferred by the major genes (R), which are often overcome by pathogens. The accumulation of the genes, which provide the resistance to a specific race of a pathogen, together with the introduction of race-non-specific resistance genes is the most effective strategy aimed at preventing the breakdown of genetically conditioned immunity. PCR markers improved the productivity and accuracy of classical plant breeding by means of marker-assisted selection (MAS). Multiplexing assays provide increased throughput, reduced reaction cost, and conservation of limited sample material, which are beneficial for breeding purposes. Here, we described the process of customizing multiplex PCR assay for the simultaneous identification of the major leaf rust resistance genes Lr19, Lr24, Lr26, and Lr38, as well as the slow rusting, race-nonspecific resistance genes: Lr34 and Lr68, in thirteen combinations. The adaptation of PCR markers for multiplex assays relied on: (1) selection of primers with an appropriate length; (2) selection of common annealing/extension temperature for given primers; and (3) PCR mixture modifications consisting of increased concentration of primers for the scanty band signals or decreased concentration of primers for the strong bands. These multiplex PCR protocols can be integrated into a marker-assisted selection of the leaf rust-resistant wheat genotypes.

Molecular analyses in wheat and Aegilops tauschii reveal a new orthologue of the leaf rust resistance gene Lr19 on chromosome 7DL of Ae. tauschii

AbstractLeaf rust is one of the most devastating wheat diseases worldwide, to which many resistance genes have been successfully introgressed from wheat wild relatives. Though the Thinopyrum ponticum‐derived leaf rust resistance gene Lr19 is widely effective, and previous studies have shown its likely presence in Aegilops tauschii, no thorough investigation has been conducted to confirm this. The current study examined the presence of Lr19 in Ae. tauschii using molecular and bioinformatic analyses. Accordingly, the wheat cultivar Thatcher was used as a susceptible control, and the Lr19‐carrying genotypes; Thatcher + Lr19 (TcLr19) and Agatha were used as resistant controls. Phenotyping using pathotype CDHLQ accompanied with genotyping using two DNA markers verified previous results of the likely presence of an orthologue for Lr19 on chromosome 7DL of Ae. tauschii (AtLr19). Sequencing of the STS GB marker products from Ae. tauschii and TcLr19 showed 99% homology between the corresponding fragments, confirming the phenotyping and genotyping results. Both sequenced fragments matched to the melatonin biosynthesis gene, O‐methyltransferase‐2 (OMT2). A hierarchical gene network was reconstructed using all identified putative genes within a genomic region containing 2.5 cM upstream and downstream of OMT2. Results indicated that numerous important biotic stress‐responsive genes such as RPM1, RGA2, TRIUR3, BURP12 and myosin‐11, were located downstream of melatonin as a master regulator molecule through the OMT2 node. To our knowledge, this is the first report of the identification of an orthologue for Lr19 in Ae. tauschii, which provides insights into the possible regulatory route of this gene.

Marker-assisted pyramiding of Thinopyrum-derived leaf rust resistance genes Lr19 and Lr24 in bread wheat variety HD2733.

This study was undertaken to pyramid two effective leaf rust resistance genes (Lr19 and Lr24) derived from Thinopyrum (syn. Agropyron), in the susceptible, but agronomically superior wheat cultivar HD2733 using marker-assisted selection. In the year 2001, HD2733 was released for irrigated timely sown conditions of the north eastern plains zone (NEPZ) of India became susceptible to leaf rust, a major disease of the region. Background selection helped in developing near-isogenic lines (NILs) of HD2733 with Lr19 and Lr24 with 97.27 and 98.94%, respectively, of genomic similarity with the parent cultivar, after two backcrossing and one generation of selfing.NILs were intercrossed to combine the genes Lr19 and Lr24. The combination of these two genes in the cultivar HD2733 is expected to provide durable leaf rust resistance in farmers' fields.

The Potential of Lr19 and Bdv2 Translocations to Improve Yield and Disease Resistance in the High Rainfall Wheat Zones of Australia

Chromosomal translocations in wheat derived from alien species are a valuable source of genetic diversity that have provided increases in resistance to various diseases and improved tolerance to abiotic stresses in wheat. These alien genomic segments can also affect multiple traits, with a concomitant ability to alter yield potential in either a positive or negative fashion. The aim of this work was to characterize the effects on yield of two types of translocations, namely T4-derived translocations from Thinopyrum ponticum, carrying the leaf rust resistance gene Lr19, and the TC14 translocation from Th. intermedium, carrying the barley yellow dwarf virus resistance gene Bdv2, in Australian adapted genetic backgrounds and under Australian conditions. A large range of germplasm was developed by crossing donor sources of the translocations into 24 Australian adapted varieties producing 340 genotypes. Yield trials were conducted in 14 environments to identify effects on yield and yield components. The T4 translocations had a positive effect on yield in one high yielding environment, but negatively affected yield in low-yielding environments. The TC14 translocation was generally benign, however, it was associated with a negative impact on yield and reduced height in two genetic backgrounds. The translocation was also associated with a delayed maturity in several backgrounds. The T4 translocations results were consistent with previously published data, whilst this is the first time that such an investigation has been undertaken on the TC14 translocation. Our data suggests a limited role for each of these translocations in Australia. The T4 translocations may be useful in high yielding environments, such as under irrigation in NSW and in the more productive high rainfall regions of south-eastern Australia. Traits associated with the TC14 translocation, such as BYDV resistance and delayed maturity, would make this translocation useful in BYDV-prone areas that experience a less pronounced terminal drought (e.g., south-eastern Australia).

PCR-based molecular markers linked to the leaf rust resistance gene Lr19 in different bread wheat cultivars

61 varieties of wheat collected in the gene fund of the Research Institute of Crop Husbandry were screened using SCAR-markers associated with the gene of resistance to brown leaf rust, Lr19. As a result of PCR analysis using SCS123 marker the 737 bp locus was detected in 48 genotypes. The expected fragment of the 688 bp was detected in 53 genotypes using the SCS253 marker. The results obtained using both markers indicate that the Lr19 gene is present on 7D chromosomes of 45 genotypes. The existence of the Lr19 gene has not been proven only for 5 from the 61 analyzed wheat genotypes.

A genetic map of Lophopyrum ponticum chromosome 7E, harboring resistance genes to Fusarium head blight and leaf rust

The leaf rust resistance gene Lr19 and Fusarium head blight (FHB) resistance quantitative trait loci (QTL) derived from the wild wheatgrass Lophopyrum ponticum have been located on chromosome 7E. The main objectives of the present study were to develop a genetic map of chromosome 7E and map the two resistance loci using a population of 237 F(7:8) recombinant inbred lines (RILs) derived from a cross between two Thatcher-L. ponticum substitution lines, K11463 (7el(1)(7D)) and K2620 (7el(2)(7D)). 532 G-SSR, E-SSR and STS markers from wheat chromosome group 7 were screened in the parent lines. Of these, 118 markers were polymorphic, with a polymorphism frequency of 22.2%. A genetic map of L. ponticum chromosome 7E was constructed with 64 markers, covering 95.76 cM, with an average genetic distance of 1.47 cM between markers. The major FHB resistance locus, temporarily assigned as FhbLoP, was mapped to the very distal region of the long arm of chromosome 7E within a 3.71 cM interval flanked by Xcfa2240 and Xswes19, which accounts for 30.46% of the phenotypic variance. Lr19 was bracketed by Xwmc273 and XBE404744, with a map distance of 1.54 and 1.43 cM from either side, respectively. The closely linked markers identified in this study will be helpful for marker-assisted introgression of the L. ponticum-derived FhbLoP and Lr19 genes into elite cultivars of wheat, and the development of a genetic map will accelerate the map-based cloning of these two genes.

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 F(2) 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 F(2) derived F(3) 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 S265(512) and S253(737) which flanked the locus Lr19 were converted to sequence characterized amplified region markers SCS265(512) and SCS253(736), respectively. The marker SCS265(512) was linked with Lr19 in a coupling phase and the marker SCS253(736) 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.

A SSR Marker for Leaf Rust Resistance Gene Lr19 in Wheat

Microsatellite was carried out in Thatcher, six near-isogenic lines and F 2 progeny of TcLr19xThatcher to develop molecular markers for leaf rust resistance gene Lr19. Thirteen primer pairs were screened, of which one primer pair Xgwm44 displayed polymorphsim in the population of TcLr19, Thatcher, and their F 2 generations. One marker closed linked to Lr19 resistance trait was obtained, and was named Xgwm44 139 bp with the genetic distance 0.9 cM. The research shows that Lr19 has more potential in marker-assisted breeding programs in wheat and provides a step stone for mapping genetic map, physical map and the eventual cloning.

Development of wheat genotypes possessing a combination of leaf rust resistance genes Lr19 and Lr24

Endopeptidase allele Ep-D1c and DNA marker-assisted selection have been used for the incorporation of Lr19 + Lr24 leaf rust resistance genes combination into adapted commercial winter wheat cultivars. The first step was the transfer of the gene Lr19 from the donor cultivar Agrus into acceptor cultivars Simona and Lívia. The progenies possessing the null allele Ep-D1c linked to the gene Lr19 have been screened for their resistance to leaf rust by isolate 4332 SaBa. The plants homozygous properties at the Ep-D1c locus and resistant against leaf rust were used for crossing with NIL Thatcher/Lr24 – a donor of the gene Lr24. Plants possessing both Lr genes were selected from F<sub>2</sub> population by STS and isozyme markers linked to the Lr genes. Progenies of 18 F<sub>2</sub> plants have been selected by STS marker and tested for resistance against leaf rust. Results obtained with isozyme and STS markers corresponded with resistance testing. Altogether 6 progenies of F<sub>3</sub> generation possessing a resistance gene combination of Lr19 + Lr24 in a homozygous condition were developed.

Identification of a molecular marker linked to an Agropyron elongatum‐derived gene Lr19 for leaf rust resistance in wheat

AbstractThe leaf rust resistance gene Lr19, transferred from Agropyron elongatum into wheat (Triticum aestivum L.) imparts resistance to all pathotypes of leaf rust (Puccinia recondita f.sp. tritici) in South‐east Asia. A segregating F2 population from a cross between the leaf rust resistant parent ‘HW 2046’ carrying Lr19 and a susceptible parent ‘Agra Local’ was screened in the phytotron against a virulent pathotype 77‐5 of leaf rust with the objective of identifying the molecular markers linked to Lr19. The gene was first tagged with a randomly amplified polymorphic DNA (RAPD) marker S73728. The RAPD marker linked to the gene Lr19 which mapped at 6.4 ± 0.035 cM distance, was converted to a sequence characterized amplified region (SCAR) marker. The SCAR marker (SCS73719) was specific to Lr19 and was not amplified in the near‐isogenic lines (NILs) carrying other equally effective alien genes Lr9, Lr28 and Lr32 enabling breeders to pyramid Lr19 with these genes.

Marker-assisted selection for leaf rust resistance in wheat by transfer of gene Lr19

Cultivar Agrus, possessing a chromosomal substitution, and cultivar Sunnan, possessing a translocation from Thinopyrum ponticum (= Agropyron elongatum, 2n = 10x) with leaf rust resistance gene Lr19 against Puccinia triticina, were crossed with the susceptible winter wheat cultivars Sofia, Simona and Lívia to transfer Lr19 into agronomi­cally better genotypes by marker-assisted selection. Altogether 304 individuals of the F2 progeny were screened for endopeptidase phenotypes. We found null endopeptidase allele Ep-D1c (marker tightly liked with resistance gene Lr19) in 49 plants. The progenies of 40 plants of the F2 generation (with Ep-D1c) were reselected with the same marker and tested for leaf rust reaction. Results achieved with the isozyme marker corresponded with those of the resistance tests. We obtained 28 F3 families with resistance gene Lr19 confirmed by presence of the null endopeptidase allele and by tests for leaf rust reaction. Field tests showed that Agrus increased the height of plants in the progenies, and the smallest negative effect on yield components was observed in both crosses with cultivar Sunnan.  

Wheat chromosome engineering at the 4x level: the potential of different alien gene transfers into durum wheat

With the aim of making the point on feasibility and relative success of alien transfers into durum wheat via chromosome engineering, three transfer works, differing in origin and content of the alien introduction and in the transfer strategy adopted, are described. For the transfer of a powdery mildew resistance gene, Pm13, originating from Aegilops longissima and previously transferred to common wheat chromosome 3B, as well as for that of the leaf rust resistance gene Lr19 and its associated Yp (yellow pigment) gene, deriving from Ag. elongatum and introduced into 7A, the common wheat recombinants were employed as donors, from which the alien segments were homologously transferred into durum genotypes. On the other hand, for the transfer of common wheat chromosome ID seed storage protein genes, ph1 mediated homoeologous recombination was repeatedly induced. This resulted in loss of individuals, including potentially desirable recombinants, probably due to imbalances created by the ph1 condition. However, recovered Gli-D1/Glu-D3 tetraploid recombinants exhibited normal transmission and fertility. Preliminary evidence indicates a normal behaviour also for Glu-D1 ‘5+10’ putative recombinants. Similarly, there was no negative impact from the transfer of the Pm13 gene, which has been successfully pyramided into Pm4a durum varieties. On the contrary, transfer of the Ag. elongatum segment showed normal female but almost no male transmission in one durum genotype. This in spite of the fact that the alien segment, proved to be, through in situ hybridization, considerably longer than previously believed, should contain an Sd-1 gene, causing preferential transmission in common wheat. While its behaviour is being checked in other durum genotypes, shortening of the alien segment, through ph1 induced recombination, is also being carried out. Possible causes of the severe negative selection that this alien transfer seemingly encounters at the tetraploid level are discussed.

Genetic control for resistance to leaf rust in wheat-Agropyron lines: Agro 139 and Agro 58

Leaf rust resistance lines of Triticum aestivum carry highly effective Lr genes from Agropyron intermedium (Host) Beauv. This Agro 58 and Agro 139 resistance segregated independently of Agropyron leaf-rust resistance genes Lr-19, Lr-24 and Lr-9 from Ae. umbellulata. Monosomic analysis showed that the Lr gene in Agro 139 was incorporated into wheat chromosome 6D. C-banding analysis could not determine the C-banding pattern of A. intermedium in wheat -Agropyron lines Agro 58 and Agro 139. It is assumed that the transfers occurred from the euchromatin regions of the Agropyron chromosomes to the euchromatin regions of the wheat chromosomes. It is suggested that the Lr gene from Agro 139 be designated LrAg (i)-1 and the Lr gene from Agro 58 designated LrAg (i)-2.