Leaf Rust Resistance Genes Research Articles

A Combination of Leaf Rust Resistance Genes, Including Lr34 and Lr46, Is the Key to the Durable Resistance of the Canadian Wheat Cultivar, Carberry.

The hexaploid spring wheat cultivar, Carberry, was registered in Canada in 2009, and has since been grown over an extensive area on the Canadian Prairies. Carberry has maintained a very high level of leaf rust (Puccinia triticina Eriks.) resistance since its release. To understand the genetic basis of Carberry’s leaf rust resistance, Carberry was crossed with the susceptible cultivar, Thatcher, and a doubled haploid (DH) population of 297 lines was generated. The DH population was evaluated for leaf rust in seven field environments at the adult plant stage. Seedling and adult plant resistance (APR) to multiple virulence phenotypes of P. triticina was evaluated on the parents and the progeny population in controlled greenhouse studies. The population was genotyped with the wheat 90 K iSelect single nucleotide polymorphism (SNP) array, and quantitative trait loci (QTL) analysis was performed. The analysis using field leaf rust response indicated that Carberry contributed nine QTL located on chromosomes 1B, 2B (2 loci), 2D, 4A, 4B, 5A, 5B, and 7D. The QTL located on 1B, 2B, 5B, and 7D chromosomes were observed in two or more environments, whereas the remainder were detected in single environments. The resistance on 1B, detected in five environments, was attributed to Lr46 and on 7D, detected in seven environments to Lr34. The first 2B QTL corresponded with the adult plant gene, Lr13, while the second QTL corresponded with Lr16. The seedling analysis showed that Carberry carries Lr2a, Lr16, and Lr23. Five epistatic effects were identified in the population, with synergistic interactions being observed for Lr34 with Lr46, Lr16, and Lr2a. The durable rust resistance of Carberry is attributed to Lr34 and Lr46 in combination with these other resistance genes, because the resistance has remained effective even though the P. triticina population has evolved virulent to Lr2a, Lr13, Lr16, and Lr23.

Identification of leaf rust resistance genes in the new Russian varieties of common wheat

Background.Wheat leaf rust caused byPuccinia triticinaErikss. is a significant wheat disease in all regions of the Russian Federation. The genetic diversity of the cultivated wheat varieties regarding the type of resistance and genes that control it ensures reliable protection of this crop against the pathogen. The aim of this work was to characterize the diversity of new Russian varieties of winter and spring common wheat for leaf rust resistance genes (Lr-genes).Materials and Methods.The research material was represented by 43 varieties of winter and 25 of spring wheat included in the State Register of Selection Achievements of the Russian Federation in 2018-2020.Results.Using molecular markers, 18Lrgenes were identified:Lr1, Lr3, Lr9, Lr10, Lr19, Lr20, Lr21, Lr24, Lr25, Lr26, Lr28, Lr29, Lr34, Lr35, Lr37, Lr41 (39), Lr47andLr66. A phytopathological test was used to clarify the results of molecular analysis. Ninety-three percent of the studied wheat varieties were found to containLrgenes, either separately or in combinations. These were the highly and partially effective genesLr24, Lr9, andLr19, adult plant resistance genesLr34andLr37, and ineffective genesLr1, Lr3, Lr10, Lr20, andLr26. TheLr24gene has been identified for the first time in Russian varieties. The spring variety ‘Leader 80’, harboring this gene, is recommended for cultivation in the West Siberian and East Siberian regions. An effective combination ofLr9+Lr26genes, individually overcome by the pathogen, was determined in the spring cultivar ‘Silach’, highly resistant to leaf rust. TheLr9gene was found in the winter variety ‘Gerda’, which is recommended for cultivation in the North Caucasus region. Previously, the varieties withLr9were not grown in the North Caucasus. An increase in the number of leaf rust resistant accessions protected by the effective adult plant resistance geneLr37is noted among wheat varieties undergoing regional adaptation testing. Many of the identifiedLrgenes (Lr19, Lr24, Lr26, Lr34, Lr37) are linked with effectiveSrgenes (Sr25, Sr24, Sr31, Sr57, andSr38), which additionally ensures stable genetic protection of wheat against stem rust.Conclusions.The obtained information about representation ofLrgenes in wheat varieties should be used in regional breeding programs. A timely replacement of genetically protected varieties allows stabilizing the populational composition of the phytopathogen and reducing the likelihood of epiphytotics.

Phenotyping and validation of molecular markers associated with rust resistance genes in wheat cultivars in Egypt.

Thirteen Egyptian wheat cultivars were evaluated and characterized for adult plant resistance to yellow, leaf, and stem rusts. SSR markers linked to yellow, leaf and stem rust resistance genes were validated and subsequently used to identify wheat cultivars containing more than one rust resistance gene. Results of the molecular marker detection indicated that several genes, either alone or in different combinations, were present among the wheat cultivars, including Yr, Yr78 (stripe rust), Lr, Lr70 (leaf rust), Sr. Sr33, SrTA10187, Sr13, and Sr35 (stem rust), and Lr34/Yr18 and Lr49/Yr29 (leaf/stripe rust). The cultivar Sakha-95 was resistant to leaf and stem rusts, and partially resistant to stripe rust; however, this cultivar contained additional rust resistance genes (Lr, Sr and Lr/Yr). The area under the disease progress curve (AUDPC) type for the various wheat cultivars differed depending on the type of rust infection (yellow, leaf, or stem rust, indicated by Yr, Lr, and Sr). The cultivars Gem-12, Sids-14, Giza-171, and Giza-168 had AUDPC types of partial resistance and resistance. All six cultivars, however, contained additional rust resistance genes. Marker-assisted selection can be applied to improve wheat cultivars with efficient gene combinations that would directly support the development of durable resistance in Egypt. Once the expression of the resistance genes targeted in this study have been confirmed by phenotypic screening, the preferable cultivars can be used as donors by Egyptian wheat breeders. The results of this study will help breeders determine the extent of resistance under field conditions when breeding for rust resistance in bread wheat.

Wheat Apoplast-Localized Lipid Transfer Protein TaLTP3 Enhances Defense Responses Against Puccinia triticina.

Plant apoplast serves as the frontier battlefield of plant defense in response to different types of pathogens. Many pathogenesis-related (PR) proteins are accumulated in apoplastic space during the onset of plant–pathogen interaction, where they act to suppress pathogen infection. In this study, we found the expression of Triticum aestivum lipid transfer protein 3 (TaLTP3) gene was unregulated during incompatible interaction mediated by leaf rust resistance genes Lr39/41 at the early infection stage. Stable transgenic wheat lines overexpressing TaLTP3 exhibited enhanced resistance to leaf rust pathogen Puccinia triticina. Transcriptome analysis revealed that overexpression of TaLTP3 specifically activated the transcription of pathogenesis-related protein 1a (TaPR1a) and multiple plant hormone pathways, including salicylic acid (SA), jasmonic acid (JA), and auxin, in response to the infection of the model bacterial pathogen Pseudomonas syringae pv. tomato DC3000. Further investigation indicated that TaLTP3 physically associated with wheat TaPR1a protein in the apoplast. Transgenic wheat lines overexpressing TaLTP3 and TaPR1a showed higher accumulations of reactive oxygen species (ROS) during plant defense responses. All these findings suggested that TaLTP3 is involved in wheat resistance against leaf rust pathogen infection and forming a TaLTP3-TaPR1a complex in apoplast against this pathogen, which provides new insights into the functional roles of PR proteins.

Characterization and Use in Wheat Breeding of Leaf Rust Resistance Genes from Durable Varieties.

Simple SummaryWheat leaf rust is one of the most significant diseases worldwide, incited by a parasitic fungus which infects leaves, affecting grain yield. This pathogen is spread by the wind over large areas through microscopic spores. This huge number of spores favors the selection of virulent forms; therefore, there is a continuous need for new resistance genes to control this disease without fungicides. These resistant genes are naturally found in resistant wheat varieties and can be introduced by standard crosses. In this work, seven resistant genes were introduced into several commercial susceptible varieties. The selection of resistance genes was assisted by DNA markers that are close to these genes on the chromosome. Additionally, the selection of desirable traits from the commercial variety was also assisted by DNA markers to accelerate the process. In field testing, the varieties developed here were resistant to leaf rust, and suitable for commercial use.Leaf rust is one of the most significant diseases of wheat worldwide. In Argentina, it is one of the main reasons for variety replacement that becomes susceptible after large-scale use. Some varieties showed durable resistance to this disease, including Buck Manantial and Sinvalocho MA. RILs (Recombinant Inbred Lines) were developed for each of these varieties and used in genetics studies to identify components of resistance, both in greenhouse inoculations using leaf rust races, and in field evaluations under natural population infections. In Buck Manantial, the APR gene LrBMP1 was associated with resistance in field tests. In crosses involving Sinvalocho MA, four genes were previously identified and associated with resistance in field testing: APR (Adult Plant Resistance) gene LrSV1, the APR genetic system LrSV2 + LrcSV2 and the ASR (All Stage Resistance) gene LrG6. Using backcrosses, LrBMP1 was introgressed in four commercial susceptible varieties and LrSV1, LrSV2 + LrcSV2 and LrG6 were simultaneously introgressed in three susceptible commercial varieties. The use of molecular markers for recurrent parent background selection allowed us to select resistant lines with more than 80% similarity to commercial varieties. Additionally, progress towards positional cloning of the genetic system LrSV2 + LrcSV2 for leaf rust APR is reported.

Identification of Two New Loci for Adult Plant Resistance to Leaf Rust and Stripe Rust in the Chinese Wheat Variety 'Neimai 836'.

The characterization of leaf rust (caused by Puccinia triticina) and stripe rust (caused by Puccinia striiformis f. sp. tritici) resistance genes is the basis for breeding resistant wheat varieties and managing epidemics of these diseases in wheat. A cross between the susceptible wheat variety 'Apav#1' and resistant variety 'Neimai 836' was used to develop a mapping population containing 148 F5 recombinant inbred lines (RILs). Leaf rust phenotyping was done in field trials at Ciudad Obregón, Mexico, in 2017 and 2018, and stripe rust data were generated at Toluca, Mexico, in 2017 and in Mianyang, Ezhou, and Gansu, China, in 2019. Inclusive complete interval mapping (ICIM) was used to create a genetic map and identify significant resistance quantitative trait loci (QTL) with 2,350 polymorphic markers from a 15K wheat single-nucleotide polymorphism (SNP) array and simple-sequence repeats (SSRs). The pleiotropic multipathogen resistance gene Lr46/Yr29 and four QTL were identified, including two new loci, QLr.hzau-3BL and QYr.hzau-5AL, which explained 3 to 16% of the phenotypic variation in resistance to leaf rust and 7 to 14% of that to stripe rust. The flanking SNP markers for the two loci were converted to Kompetitive Allele-Specific PCR (KASP) markers and used to genotype a collection of 153 wheat lines, indicating the Chinese origin of the loci. Our results suggest that Neimai 836, which has been used as a parent for many wheat varieties in China, could be a useful source of high-level resistance to both leaf rust and stripe rust.

Using the synthetic form RS5 to obtain new introgressive lines of common wheat.

The use of the gene pool of wild relatives, which have a signif icant reserve of genetic diversity, is of immediate interest for breeding common wheat. The creation and use of synthetic forms as “bridges” is an effective method of transferring valuable genetic material from wild relatives to cultivated wheat. For this purpose, genome addition, genome substitution and recombinant “secondary” synthetic forms have been created in the P.P. Lukyanenko National Center of Grain. The synthetic recombination form RS5 (BBAASDt), in which the third genome consists of chromosomes of Aegilops speltoides (S) and Aegilops tauschii (Dt), was obtained from crossing the synthetic forms Avrodes (BBAASS) and M.it./ Ae. tauschii (BBAADt Dt), in which the D genome from Ae. tauschii was added to the BBAA genomes of the durum wheat cultivar Mutico italicum. Introgression lines resistant to leaf rust, yellow rust and powdery mildew have been obtained from backcrosses with the susceptible common wheat cultivars Krasnodarskaya 99, Rostislav and Zhirovka. Twelve resistant lines that additionally have high technological characteristics of grain and f lour have been selected. The cytological study (С-banding) has revealed chromosomal modif ications in 6 of 8 lines under study. The rearrangements mainly affected the chromosomes of the D genome, 1D, 3D, 4D, 6D and 7D. It was found that in most cases the genetic material from the synthetic form RS5 in the studied lines was represented by substituted chromosomes from Ae. tauschii. In line 5791p17, the substitution of chromosomes 6D from Ae. tauschii and 7D from Ae. speltoides was revealed. Substitutions 4D(4Dt), 6D(6Dt) from Ae. tauschii and 7D(7S) from Ae. speltoides were obtained for the f irst time. Molecular analysis of 12 lines did not reveal effective leaf rust resistance genes, presumably present in synthetic forms of M.it./Ae. tauschii and Avrodes. It is assumed that the lines may carry previously unidentif ied genes for fungal disease resistance, in particular for resistance to leaf rust, from Ae. tauschii and Ae. speltoides

Mining Middle Eastern and Central Asian Barley Germplasm to Understand Diversity for Resistance to Puccinia hordei, Causal Agent of Leaf Rust

Vast collections of barley germplasm have been established and conserved in various global gene banks. These collections hold tremendous genetic diversity for resistance genes to Puccinia hordei, a causal agent of barley leaf rust. This study was undertaken to discover, characterize and postulate the known Rph genes (resistance to Puccinia hordei) and identify novel sources of ASR (all-stage resistance) and APR (adult plant resistance) to P. hordei. A core set of 315 barley lines were rust-tested as seedlings for their response to eight Australian pathotypes of P. hordei and genotyped with molecular markers linked to the known characterised ASR and APR genes. These tests led to the postulation of ASR leaf rust resistance genes Rph1, Rph2, Rph3, Rph9.am, Rph12, Rph15, Rph19 and Rph25 singly or in combination. Field tests revealed that the vast majority of lines (84%) carried APR. Genotyping of the APR-carrying lines with markers bPb-0837, Ebmac0603 and sun43-44 identified lines that likely carry the known APR genes Rph20, Rph23 and Rph24 singly or in combination. Thirty-nine per cent of the lines were negative for all the three markers and were thus postulated to carry uncharacterized APR. The sources of resistance identified in this study provide a valuable resource to breeders for further utilization and diversifying the genetic basis of leaf rust resistance in barley.

Leaf tip necrosis and seedling leaf rust resistance conditioned by Lr34 in wheat grown at low temperatures

Wheat leaf rust, caused by Puccinia triticina Eriks., is a common and destructive disease of wheat worldwide. The durable leaf rust-resistance gene Lr34 is typically described as an adult plant resistance gene, which is not expressed at the seedling stage at normal greenhouse temperatures. However, Lr34 is expressed at the seedling stage when plants with Lr34 are grown at low temperatures, i.e. approximately 8°C. Lines and cultivars with Lr34 demonstrate a range of responses from near immunity to intermediate reactions on the primary leaf. The Thatcher-Lr34 near isogenic lines ‘RL6058ʹ (Thatcher*6/PI58548) and ‘RL6091ʹ (Tc*6/Chinese Spring) were resistant to a number of P. triticina virulence phenotypes at the seedling stage when grown under these cool conditions, but were susceptible at typical greenhouse temperatures. A double haploid population from a cross between ‘Thatcher’ and ‘RL6058ʹ, previously genotyped for Lr34, was grown at 8°C and screened for leaf rust response at the seedling stage. The population is segregated by a seedling resistance gene that corresponds to the presence of a resistance allele of Lr34. After extended incubation at cold temperatures, the seedling progeny lines with Lr34 also developed distinct leaf tip necrosis; this phenotype was absent in susceptible plants, similar to their reaction as adult plants. When uninfected plants from this population were grown to the adult plant stage at 10°C, those with the Lr34 resistance allele had extensive leaf necrosis and shrunken, sometimes sterile, spikes.

Virulence Phenotyping of Leaf Rust (Puccinia triticina) Isolates from Southern Pakistan

Collections of Puccinia triticina were made from farmers’ fields of five different agro-ecological locations (Sakrand, Tandojam, Larkana, Sanghar and Badin) of Sindh province, Pakistan from 2015 and 2016, to identify the virulence variation. Single uredinial isolates were investigated for virulence phenotyping on 24 near isogenic (Thatcher wheat) lines which differ for single Lr resistance genes. Spores from two locations (Sakrand and Tandojam) were not viable and could not be revived and only urediniospores of three locations (Larkana, Sanghar and Badin) were revived. None of the pathotypes had virulence to Thatcher wheat lines with leaf rust resistance genes Lr23 and Lr42. However, Lr24, LrB, Lr10, Lr14b and Lr20 genes exhibited susceptibility response i.e. (HITs 3 4) with all tested pathotypes. Based on virulence, ten virulence phenotypes (MSCTNS, RTSTNS, RKTRGS, PNDQDS, JDBQGJ, MDPSDS, RTPTPS, MNPSDS, MJLTGS and MSPTDS) were identified among the ten isolates, designated with six-letter code. Two phenotypes RTSTNS RTPTPS exhibited broad spectrum, both were virulent to nineteen resistance genes of leaf rust while pathotype JDBQGJ had narrow spectrum as compared to all other tested, with virulence to just eight resistance genes of leaf rust. Among the locations virulence variability of leaf rust was also recorded. Most of identified races were virulent to more than one of leaf rust resistance genes. Resistance genes (Lr42 and Lr23) identified as effective can be exploited to achieve leaf rust resistance in wheat. Further, the study provides virulence profile of the area may help to manage the leaf rust pathogen.

Three co-located resistance genes confer resistance to leaf rust and stripe rust in wheat variety Borlaug 100

Leaf rust (LR) and stripe rust (YR) are important diseases in wheat producing areas worldwide and cause severe yield losses under favorable environmental conditions when susceptible varieties are grown. We determined the genetic basis of resistance to LR and YR in variety Borlaug 100 by developing and phenotyping a population of 198 F6 recombinant inbred lines derived from a cross with the susceptible parent Apav#1. LR and YR phenotyping were conducted for 4 and 3 seasons, respectively, at CIMMYT research stations in Mexico under artificial epidemics. Mendelian segregation analyses indicated that 3–5 LR and 2 YR genes conferred resistance in Borlaug 100. Lr46/Yr29 (1BL), Yr17 (2AS) and Yr30 (3BS) were present in the resistant parent and segregated in the RIL population based on characterization by molecular markers linked to these genes. When present alone, Lr46/Yr29 caused average 13% and 16% reductions in LR and YR severities, respectively, in RILs. Similarly, Yr17 and Yr30 reduced YR severities by 57% and 11%, respectively. The Yr30 and the Yr17 translocation were also associated with 27% and 14% reductions, respectively, in LR severity, indicating that the 3BS and 2AS chromosomal regions likely carry new slow rusting LR resistance genes, temporarily designated as LrB1 and LrB2, respectively. Additive effects of Yr30*Yr17, Yr29*Yr17 and Yr29*Yr30 on YR and LR were significant and reduced YR severities by 56%, 55%, and 45%, respectively, and LR severities by 34%, 40%, and 45%, respectively. Furthermore, interaction between the three genes was also significant, with mean reductions of 70% for YR and 54% for LR severities. Borlaug 100, or any one of the 21 lines with variable agronomic traits but carrying all three co-located resistance loci, can be used as resistance sources in wheat breeding programs.

Lr21 diversity unveils footprints of wheat evolution and its new role in broad-spectrum leaf rust resistance.

Aegilops tauschii, the progenitor of the wheat D genome, contains extensive diversity for biotic and abiotic resistance. Lr21 is a leaf rust resistance gene, which did not enter the initial gene flow from Ae. tauschii into hexaploid wheat due to restrictive hybridization events. Here, we used population genetics and high-resolution comparative genomics to study evolutionary and functional divergence of Lr21 in diploid and hexaploid wheats. Population genetics identified the original Lr21, lr21-1 and lr21-2 alleles and their evolutionary history among Ae. tauschii accessions. Comparative genetics of Lr21 variants between Ae. tauschii and cultivated genotypes suggested at least two independent polyploidization events in bread wheat evolution. Further, a recent re-birth of a unique Lr21-tbk allele and its neofunctionalization was discovered in the hexaploid wheat cv. Tobak. Altogether, four independent alleles were investigated and validated for leaf rust resistance in diploid, synthetic hexaploid and cultivated wheat backgrounds. Besides seedling resistance, we uncover a new role of the Lr21 gene in conferring an adult plant field resistance. Seedling and adult plant resistance turned out to be correlated with developmentally dependent variation in Lr21 expression. Our results contribute to understand Lr21 evolution and its role in establishing a broad-spectrum leaf rust resistance in wheat.

Ne2, a typical CC-NBS-LRR-type gene, is responsible for hybrid necrosis in wheat.

Hybrid necrosis, caused by complementary genes Ne1 and Ne2, is a serious barrier for combining desirable traits from different genotypes of wheat, affecting the full utilisation of heterosis. To date, both Ne1 and Ne2 are still not isolated although they were documented decades ago. We report here the map-based cloning and functional characterisation of Ne2, encoding a coiled coil-nucleotide-binding site-leucine-rich repeat (CC-NBS-LRR) protein. Homozygous frameshift mutations generated using the CRISPR/Cas9 approach confirmed the Ne2-inducing hybrid necrosis in wheat. Upregulated expression of Ne2 induced by Ne1 and excess hydrogen peroxide accumulation are associated with the necrosis formation. Genetic analyses of a Ne2 allele (Ne2m ) and leaf rust resistance gene LrLC10/Lr13 revealed that they might be the same gene. Furthermore, we demonstrated that the frequency of the Ne2 allele was much lower in landraces (2.00%) compared with that in modern cultivars (13.62%), suggesting that Ne2 allele has been partially applied in wheat genetic improvement. Our findings open opportunities of thoroughly investigating the molecular mechanism of hybrid necrosis, selecting Lr13 and simultaneously avoiding hybrid necrosis in wheat breeding through marker-assisted selection.

Evaluation of Wheat Germplasm for Resistance to Leaf Rust (Puccinia triticina) and Identification of the Sources of Lr Resistance Genes Using Molecular Markers.

Leaf rust, caused by Puccinia triticina (Ptr), is a significant disease of spring wheat spread in Kazakhstan. The development of resistant cultivars importantly requires the effective use of leaf rust resistance genes. This study aims to: (i) determine variation in Ptr population using races from the East Kazakhstan, Akmola, and Almaty regions of Kazakhstan; (ii) examine resistance during seedling and adult plant stages; and (iii) identify the sources of Lr resistance genes among the spring wheat collection using molecular markers. Analysis of a mixed population of Ptr identified 25 distinct pathotypes. Analysis of these pathotypes using 16 Thatcher lines that are near-isogenic for leaf rust resistance genes (Lr) showed different virulence patterns, ranging from least virulent “CJF/B” and “JCL/G” to highly virulent “TKT/Q”. Most of the pathotypes were avirulent to Lr9, Lr19, Lr24, and Lr25 and virulent to Lr1, Lr2a, Lr3ka, Lr11, and Lr30. The Ptr population in Kazakhstan is diverse, as indicated by the range of virulence observed in five different races analyzed in this study. The number of genotypes showed high levels of seedling resistance to each of the five Ptr races, thus confirming genotypic diversity. Two genotypes, Stepnaya 62 and Omskaya 37, were highly resistant to almost all five tested Ptr pathotypes. Stepnaya 62, Omskaya 37, Avangard, Kazakhstanskaya rannespelaya, and Kazakhstanskaya 25 were identified as the most stable genotypes for seedling resistance. However, most of the varieties from Kazakhstan were susceptible in the seedling stage. Molecular screening of these genotypes showed contrasting differences in the genes frequencies. Among the 30 entries, 22 carried leaf rust resistance gene Lr1, and two had Lr9 and Lr68. Lr10 and Lr28 were found in three and four cultivars, respectively. Lr19 was detected in Omskaya 37. Two single cultivars separately carried Lr26 and Lr34, while Lr37 was not detected in any genotypes within this study. Field evaluation demonstrated that the most frequent Lr1 gene is ineffective. Kazakhstanskaya 19 and Omskaya 37 had the highest number of resistance genes: three and four Lr genes, respectively. Two gene combinations (Lr1, Lr68) were detected in Erythrospermum 35 and Astana. The result obtained may assist breeders in incorporating effective Lr genes into new cultivars and developing cultivars resistant to leaf rust.

Identification of known leaf rust resistance genes in bread wheat cultivars from China

Leaf rust caused by Puccinia triticina Eriks. (Pt) is one of the most devastating fungal pathogens affecting wheat (Triticum aestivum L.) production worldwide. Deployment of resistant cultivars is the most environmentally friendly approach to control the disease. In this study, thirty-seven wheat lines from the Hubei and Shaanxi provinces in China were evaluated for seedling resistance in the greenhouse using eighteen Pt races. These lines were also tested for slow rusting resistance in the field in the 2014 to 2018 growing seasons. Eleven molecular markers closely associated with known Lr genes were used as part of the postulation process. Seven known Lr genes, 1, 13, 18, 14a, 26, 34 and 46 either singly or in combination were postulated in twenty-five cultivars. Lr1 and Lr26 were the most commonly identified genes detected in thirteen and ten cultivars, respectively. Lr13 and Lr46 were each found in four and five cultivars. Lr34 was present in three cultivars. Lr18 and Lr14a were identified in cultivar Xi’nong 538. Six cultivars displayed slow rusting resistance in the field tests. The resistant cultivars identified in the present study can be used as resistance parents in crosses aimed at pyramiding and the deployment of leaf rust resistance genes in China.

Whole-genome sequencing of multiple isolates of Puccinia triticina reveals asexual lineages evolving by recurrent mutations.

The wheat leaf rust fungus, Puccinia triticina Erikss., is a worldwide pathogen of tetraploid durum and hexaploid wheat. Many races of P. triticina differ for virulence to specific leaf rust resistance genes and are found in most wheat-growing regions of the world. Wheat cultivars with effective leaf rust resistance exert selection pressure on P. triticina populations for virulent race types. The objectives of this study were to examine whole-genome sequence data of 121 P. triticina isolates and to gain insight into race evolution. The collection included isolates comprising of many different race phenotypes collected worldwide from common and durum wheat. One isolate from wild wheat relative Aegilops speltoides and two from Ae. cylindrica were also included for comparison. Based on 121,907 informative variants identified relative to the reference Race 1-1 genome, isolates were clustered into 11 major lineages with 100% bootstrap support. The isolates were also grouped based on variation in 1311 predicted secreted protein genes. In gene-coding regions, all groups had high ratios of nonsynonymous to synonymous mutations and nonsense to readthrough mutations. Grouping of isolates based on two main variation principle components for either genome-wide variation or variation just within the secreted protein genes, indicated similar groupings. Variants were distributed across the entire genome, not just within the secreted protein genes. Our results suggest that recurrent mutation and selection play a major role in differentiation within the clonal lineages.

Efficiency of a Seedling Phenotyping Strategy to Support European Wheat Breeding Focusing on Leaf Rust Resistance.

Simple SummaryLeaf rust resistance is of high importance for the European wheat production in order to avoid yield and quality losses. Modern breeding is aiming to maximize the selection gain within a short time period. To realize a successful resistance breeding, detailed knowledge about the genetic architecture of leaf rust resistance, as well as a precise and fast phenotyping strategy, are necessary. The examination of detached leaf assays of juvenile plants inoculated under controlled conditions and phenotyped by a robotic-based, high-throughput system is a promising approach in this respect. Known leaf rust resistance genes showing qualitative or quantitative effects and their expression starts at different developmental stages of wheat. Therefore, this study validated the transferability of results from detached leaf assays to the field and assessed the benefits of this phenotyping strategy to support leaf rust resistance breeding. Phenotyping detached leaves of wheat seedlings by using an automated, high-throughput methodology is a valuable tool to improve leaf rust resistance.Leaf rust resistance is of high importance for a sustainable European wheat production. The expression of known resistance genes starts at different developmental stages of wheat. Breeding for resistance can be supported by a fast, precise, and resource-saving phenotyping. The examination of detached leaf assays of juvenile plants inoculated under controlled conditions and phenotyped by a robotic- and computer-based, high-throughput system is a promising approach in this respect. Within this study, the validation of the phenotyping workflow was conducted based on a winter wheat set derived from Central Europe and examined at different plant developmental stages. Moderate Pearson correlations of 0.38–0.45 comparing leaf rust resistance of juvenile and adult plants were calculated and may be mainly due to different environmental conditions. Specially, the infection under controlled conditions was limited by the application of a single rust race at only one time point. Our results suggest that the diversification with respect to the applied rust race spectrum is promising to increase the consistency of detached leaf assays and the transferability of its results to the field.

Mining of Leaf Rust Resistance Genes Content in Egyptian Bread Wheat Collection.

Wheat is a major nutritional cereal crop that has economic and strategic value worldwide. The sustainability of this extraordinary crop is facing critical challenges globally, particularly leaf rust disease, which causes endless problems for wheat farmers and countries and negatively affects humanity’s food security. Developing effective marker-assisted selection programs for leaf rust resistance in wheat mainly depends on the availability of deep mining of resistance genes within the germplasm collections. This is the first study that evaluated the leaf rust resistance of 50 Egyptian wheat varieties at the adult plant stage for two successive seasons and identified the absence/presence of 28 leaf rust resistance (Lr) genes within the studied wheat collection. The field evaluation results indicated that most of these varieties demonstrated high to moderate leaf rust resistance levels except Gemmeiza 1, Gemmeiza 9, Giza162, Giza 163, Giza 164, Giza 165, Sids 1, Sids 2, Sids 3, Sakha 62, Sakha 69, Sohag 3 and Bany Swif 4, which showed fast rusting behavior. On the other hand, out of these 28 Lr genes tested against the wheat collection, 21 Lr genes were successfully identified. Out of 15 Lr genes reported conferring the adult plant resistant or slow rusting behavior in wheat, only five genes (Lr13, Lr22a, Lr34, Lr37, and Lr67) were detected within the Egyptian collection. Remarkedly, the genes Lr13, Lr19, Lr20, Lr22a, Lr28, Lr29, Lr32, Lr34, Lr36, Lr47, and Lr60, were found to be the most predominant Lr genes across the 50 Egyptian wheat varieties. The molecular phylogeny results also inferred the same classification of field evaluation, through grouping genotypes characterized by high to moderate leaf rust resistance in one cluster while being highly susceptible in a separate cluster, with few exceptions.

Fine Mapping of the Leaf Rust Resistance Gene Lr65 in Spelt Wheat 'Altgold'.

Wheat leaf rust (also known as brown rust), caused by the fungal pathogen Puccinia triticina Erikss. (Pt), is one by far the most troublesome wheat disease worldwide. The exploitation of resistance genes has long been considered as the most effective and sustainable method to control leaf rust in wheat production. Previously the leaf rust resistance gene Lr65 has been mapped to the distal end of chromosome arm 2AS linked to molecular marker Xbarc212. In this study, Lr65 was delimited to a 0.8 cM interval between flanking markers Alt-64 and AltID-11, by employing two larger segregating populations obtained from crosses of the resistant parent Altgold Rotkorn (ARK) with the susceptible parents Xuezao and Chinese Spring (CS), respectively. 24 individuals from 622 F2 plants of crosses between ARK and CS were obtained that showed the recombination between Lr65 gene and the flanking markers Alt-64 and AltID-11. With the aid of the CS reference genome sequence (IWGSC RefSeq v1.0), one SSR marker was developed between the interval matched to the Lr65-flanking marker and a high-resolution genetic linkage map was constructed. The Lr65 was finally located to a region corresponding to 60.11 Kb of the CS reference genome. The high-resolution genetic linkage map founded a solid foundation for the map-based cloning of Lr65 and the co-segregating marker will facilitate the marker-assisted selection (MAS) of the target gene.

Linkage of Lr55 wheat leaf rust resistance gene with microsatellite and DArT-based markers

Every year, leaf rust causes losses of wheat yield and quality all around the world. Breeding of cereals resistant to fungal diseases offers a long-term alternative for chemical protection and becomes increasingly important for organic farming and ecological food production. Accumulation of effective resistance genes in a single genotype requires recurrent enrichment of the wheat genepool with promising resistance alleles marked with DNA tags. Lr55 gene was transferred to KS04WGRC45 wheat from Elymus trachycaulus and provides effective resistance to leaf rust. Two F2 mapping populations Bogatka × KS04WGRC45 (BK) and Nadobna × KS04WGRC45 (NK) were developed in order to identify the molecular marker(s) linked to the Lr55 gene. In total the segregations of 22 microsatellite and 7 DArT-based markers were used to create linkage groups corresponding to 1B chromosomes in individual mapping populations. At consensus map, Lr55 gene was flanked by XGwm374 and XWmc406 markers at distance of 8.3 and 14.8 cM, respectively. Until now, no molecular markers have been reported for Lr55 gene, and the molecular markers developed may be a starting point for the selection of plant materials for the presence of the Lr55 gene. However, additional marker systems based on next-generation sequencing may be necessary for more accurate location of Lr55 gene location.