Phytopathological and molecular genetic identification of leaf rust resistance genes in common wheat accessions with alien genetic material

Leaf rust resistance genes were sought in 23 resistant common wheat accessions with alien genetic material of Aegilops speltoides, Ae. triuncialis, and Triticum kiharae from the Arsenal collection. The genes were identified by common phytopathological tests and PCR analysis with STS markers linked with the known Lr genes. None of the methods identified the resistance genes in two accessions. In the other accessions, the combination of the two methods broadened the spectrum of detectable genes and, in some cases, allowed double verification of the presence of a resistance gene. Most accessions proved to contain several leaf rust resistance genes, combining juvenile and adult plant ones. The accessions were found to contain gene combinations that ensured field resistance (Lr13 + Lr10 and Lr12 + Lr34) and immunity under the conditions of the Non-Chernozem region. Accessions with alien genetic material contained a unique combination of five or six resistance genes. Since the accessions were rich in leaf rust resistance genes, including effective ones, and carried rare combinations of these genes, they were proposed as donors to be universally employed in breeding for immunity in all regions of Russia.     

Leaf rust resistance genes of wheat: identification in cultivars and resistance sources

Thirty-seven wheat cultivars originating from seven European countries were examined by using sequence tagged site (STS) markers for seven Lr (leaf rust = brown rust) resistance genes against the fungal pathogen of wheat Puccinia recondita f. sp. tritici (Lr9, Lr10, Lr19, Lr24, Lr26 and Lr37). Additionally, 22 accessions with various Lr genes from two germplasm collections were tested. A Scar (sequence-characterized amplified region) marker for Lr24 and a CAPS (Cleaved Amplified Polymorphic Sequence) marker for Lr47 were also used to identify those genes in the wheat accessions. Each marker amplified one specific DNA fragment. Three Lr gene markers were identified in wheat cultivars (Lr10, Lr26 and Lr37). Another four markers (Lr9, Lr19, Lr24 and Lr47) were found in breeding lines carrying leaf rust resistance genes. The results were compared with leaf rust resistance gene postulations made in previous studies, based on multipathotype testing. Markers for Lr10, Lr26 and Lr37 may be useful in marker-assisted breeding.     

Resistance of spring wheat cultivars and lines to leaf rust

Spring wheat nursery accessions, including 18 spring wheat lines derived in CIMMYT, Mexico, and 12 spring wheat cultivars bred in Poland, along with cultivars Frontana and Sumai 3 as resistant controls, were examined for resistance to leaf rust under field conditions. Multipathotype tests with 16 different pathogen isolates were performed for postulation of Lr genes in Polish cultivars. Besides, STS markers for resistance genes Lr1, Lr9, Lr10, Lr24, Lr28, Lr37 were analysed in the studied cultivars and lines with Thatcher near-isogenic lines as positive controls. All Polish cultivars appeared to be susceptible to leaf rust. Ten of the CIMMYT nursery lines (IPG-SW: #7, 11, 14, 21, 22, 23, 27, 29, 30, 32) and cv. Frontana were resistant in the same environment and can be sources of resistance genes. Marker for the Lr10 gene was identified in 6 accessions (IPG-SW #14, 22, 23, 29, 30, 32) exhibiting resistance to leaf rust, whereas markers for Lr1 and Lr28 genes were observed in all the examined accessions. STS markers for Lr9, Lr24 and Lr37 genes were not identified in the investigated accessions.     

Genetic studies of leaf and stem rust resistance in six accessions of Triticum turgidum var. dicoccoides.

Six accessions of Triticum turgidum var. dicoccoides L. (4x, AABB) of diverse origin were tested with 10 races of leaf rust (Puccinia recondita f.sp. tritici Rob. ex Desm.) and 10 races of stem rust (P. graminis f.sp. tritici Eriks. &Henn.). Their infection type patterns were all different from those of lines carrying the Lr or Sr genes on the A or B genome chromosomes with the same races. The unique reaction patterns are probably controlled by genes for leaf rust or stem rust resistance that have not been previously identified. The six dicoccoides accessions were crossed with leaf rust susceptible RL6089 durum wheat and stem rust susceptible 'Kubanka' durum wheat to determine the inheritance of resistance. They were also crossed in diallel to see whether they carried common genes. Seedlings of F1, F2, and BC1F2 generations from the crosses of the dicoccoides accessions with RL6089 were tested with leaf rust race 15 and those from the crosses with 'Kubanka' were tested with stem rust race 15B-1. The F2 populations from the diallel crosses were tested with both races. The data from the crosses with the susceptible durum wheats showed that resistance to leaf rust race 15 and stem rust race 15B-1 in each of the six dicoccoides accessions is conferred by a single dominant or partially dominant gene. In the diallel crosses, the dominance of resistance appeared to be affected by different genetic backgrounds. With one exception, the accessions carry different resistance genes: CI7181 and PI 197483 carry a common gene for resistance to leaf rust race 15. Thus, wild emmer wheat has considerable genetic diversity for rust resistance and is a promising source of new rust resistance genes for cultivated wheats.     

8个小麦育种亲本抗叶锈基因分析

选取19个小麦叶锈菌生理小种对8个小麦育种亲本进行成株期和苗期抗叶锈病鉴定及基因推导,同时利用与24个抗叶锈基因紧密连锁或共分离的31个分子标记进行分子检测。推测出L83#-5与L83#-6含有Lr1,可能含有Lr2c和Lr42;L/PL2003-1含有Lr1,可能含有Lr2c、Lr28和Lr42;贵农13号可能含有Lr28;92R137可能含有Lr2c和Lr28;L201含有Lr1,可能含有Lr2c、Lr16和Lr28;TM可能含有Lr41和其他抗叶锈基因。研究结果表明,测试的8个小麦育种亲本中TM的抗叶锈性最好,具有很好的抗叶锈病应用潜力,可作为小麦抗叶锈病育种的重要抗源。     

Genetic control of effective juvenile resistance to leaf rust in collection accessions of Triticum aestivum L

Of 153 accessions reported to be resistant to leaf rust (Puccinia recondita Rob. ex. Desm.), only 70 were not affected by a pooled P. recondita population. According to phytopathological tests (inoculation with test clones), 14 accessions contained the Lr19 gene; 36, the Lr24 gene; 1, the Lr41 gene; and 19 presumably had the Lr9 gene. The presence of these resistance genes was confirmed by hybrid analysis for 26 accessions. Of 28 accessions reported to carry new effective resistance genes other than the known genes, 23 were affected by the P. recondita population. In four of the other five accessions, resistance proved to be controlled by known genes. Possible causes of false identification of new effective leaf rust resistance genes in wheat are discussed.     

Resistance to fungal diseases in hybrid progeny from crosses between common wheat variety Saratovskaia 29 and the amphidiploid Triticum timopheevii/Triticum tauschii (AAGGDD)

The progeny of BC6F2-BC9F(2)-4 has been analyzed for resistance to brown rust (Lr genes) and powdery mildew (Pm genes). This progeny was obtained due to introgression of the alien material from the synthetic hexaploid wheat Triticum timopheevii/Aegilops squarrosa (= Triticum tauschii AAGGDD, 2n = 42) into the common wheat variety Saratovskaya 29. Against the background of natural infection, the lines resistant to both diseases and to either of them were developed. The brown-rust and powdery-mildew resistance is controlled by one/two effective independent genes Lr and Pm.     

Marker-assisted selection for leaf rust resistance genes Lr19 and Lr24 in wheat (Triticum aestivum L.)

Leaf rust caused by Puccinia recondita f.sp. tritici is a wheat disease of worldwide importance. Wheat genotypes known to carry specific rust resistance genes and segregating lines that originated from various cross combinations and derived from distinct F2 lineage, so as to represent a diverse genetic background, were included in the present study for validation of molecular markers for Lr19 and Lr24. STS markers detected the presence of the leaf rust resistance gene Lr19 in a Thatcher NIL (Tc*Lrl9) and Inia66//CMH81A575 and of the gene Lr24 in the genotypes Arkan, Blue Boy II, Agent and CI 17907. Validation of molecular markers for Lr19 and Lr24 in parental lines, followed by successful detection of these genes in F3 lines from various cross combinations, was carried out. The molecular test corresponded well with the host-pathogen interaction test response of these lines.     

Association mapping of leaf rust resistance loci in a spring wheat core collection

Key message

We identified 15 potentially novel loci in addition to previously characterized leaf rust resistance genes from 1032 spring wheat accessions. Targeted AM subset panels were instrumental in revealing interesting loci.

Abstract

Leaf rust is a common disease of wheat, consistently reducing yields in many wheat-growing regions of the world. Although fungicides are commonly applied to wheat in the United States (US), genetic resistance can provide less expensive, yet effective control of the disease. Our objectives were to map leaf rust resistance genes in a large core collection of spring wheat accessions selected from the United States Department of Agriculture-Agricultural Research Service National Small Grains Collection (NSGC), determine whether previously characterized race-nonspecific resistance genes could be identified with our panel, and evaluate the use of targeted panels to identify seedling and adult plant resistance (APR) genes. Association mapping (AM) detected five potentially novel leaf rust resistance loci on chromosomes 2BL, 4AS, and 5DL at the seedling stage, and 2DL and 7AS that conditioned both seedling and adult plant resistance. In addition, ten potentially novel race-nonspecific resistance loci conditioned field resistance and lacked seedling resistance. Analyses of targeted subsets of the accessions identified additional loci not associated with resistance in the complete core panel. Using molecular markers, we also confirmed the presence and effectiveness of the race-nonspecific genes Lr34, Lr46, and Lr67 in our panel. Although most of the accessions in this study were susceptible to leaf rust in field and seedling tests, many resistance loci were identified with AM. Through the use of targeted subset panels, more loci were identified than in the larger core panels alone.

     

Molecular and phenotypic characterization of seedling and adult plant leaf rust resistance in a world wheat collection

Genetic resistance is the most effective approach to managing wheat leaf rust. The aim of this study was to characterize seedling and adult plant leaf rust resistance of a world wheat collection. Using controlled inoculation with ten races of Puccinia triticina, 14 seedling resistance genes were determined or postulated to be present in the collection. Lr1, Lr3, Lr10 and Lr20 were the most prevalent genes around the world while Lr9, Lr14b, Lr3ka and/or Lr30 and Lr26 were rare. To confirm some gene postulations, the collection was screened with gene-specific molecular markers for Lr1, Lr10, Lr21 and Lr34. Although possessing the Lr1 and/or Lr10 gene-specific marker, 51 accessions showed unexpected high infection types to P. triticina race BBBD. The collection was tested in the field, where rust resistance ranged from nearly immune or highly resistant with severity of 1 % and resistant host response to highly susceptible with severity of 84 % and susceptible host response. The majority of the accessions possessing the adult plant resistance (APR) gene Lr34 had a maximum rust severity of 0–35 %, similar to or better than accession RL6058, a Thatcher-Lr34 near-isogenic line. Many accessions displayed an immune response or a high level of resistance under field conditions, likely as a result of synergy between APR genes or between APR and seedling resistance genes. However, accessions with three or more seedling resistance genes had an overall lower field severity than those with two or fewer. Immune or highly resistant accessions are potential sources for improvement of leaf rust resistance. In addition, some lines were postulated to have known but unidentified genes/alleles or novel genes, also constituting potentially important sources of novel resistance.     

Leaf rust (Puccinia triticina) resistance in wheat (Triticum aestivum) cultivars grown in Northern Europe 1992-2002

Diversity of resistance to leaf rust caused by Puccinia triticina can be enhanced in wheat (Triticum aestivum) cultivars through a better knowledge of resistance genes that are present in important cultivars and germplasm. Multi-pathotype tests on 84 wheat cultivars grown in Denmark, Finland, Norway and Sweden during 1992-2002 and 39 differential testers enabled the postulation of nine known genes for seedling resistance to leaf rust. Genes Lr1, Lr2a, Lr3, Lr10, Lr13, Lr14a, Lr17, Lr23 and Lr26 were found singly or in combination in 47 of the cultivars (55.9%). The most frequently occurring genes in cultivars grown in Sweden were Lr13 (20.4%), Lr14a (14.8%) and Lr26 (14.8%). Lr14a was the most common gene in cultivars grown in Norway (18.7%), Lr13 in Denmark (35.5%) and Lr10 in Finland (20.0%). Although 28 cultivars (33.3%) exhibited a response pattern that could not be assigned to resistance genes or combinations present in the tester lines, several pathotypes carried virulence and hence these genes or combinations are of limited use. Nine cultivars (10.7%) lacked detectable seedling resistance. One cultivar was resistant to all pathotypes used in the study.     

Genetic research as the valid base of strategies for breeding rust resistant wheats

It is known that few wheat cultivars maintain their resistance to rust diseases for a long period of time, particularly when crop populations become genetically more uniform. A number of genetically diverse, so far unexploited, sources of rust resistance in the natural as well as mutagenized population of wheat cultivars were identified. Several of these genes were placed in agronomically superior well-adapted backgrounds so that they could be used as pre-breeding stocks for introducing genetic diversity for resistance in a crop population. Some of these stocks when employed as parents in several cross combinations in a breeding programme have generated a number of promising cultivars with diversity for resistance.Many presently grown wheats in India, near-isogenic lines each with Lr14b, Lr14ab, Lr30 and certain international cultivars were identified as possessing diverse sources of adult plant resistance (APR) to leaf rust. Prolonged leaf rust resistance in some of the Indian cultivars was attributed to the likely presence of Lr34 either alone or in combination with other APR components. Tests of allelism carried out in certain cultivars that continue to show adequate levels of field resistance confirm the presence of Lr34, which explains the role that this gene has played in imparting durability for resistance to leaf rust. Also, Lr34 in combination with other APR components increases the levels of resistance, which suggests that combination of certain APR components should be another important strategy for breeding cultivars conferring durable and adequate levels of resistance. A new adult plant leaf rust resistance source that seems to be associated with durability in Arjun has been postulated. Likewise, cultivars possessing Sr2 in combination with certain other specific genes have maintained resistance to stem rust.Further, non-specific resistances that were transferred across widely different genotypes into two of the popular Indian wheats provided easily usable materials to the national breeding programmes for imparting durable resistance to stripe rust.     

15个小麦农家品种抗叶锈基因分析

本研究旨在明确小麦农家品种中可能含有的抗叶锈病基因,为抗源的选择和利用提供理论依据。以15个小麦农家品种、感病对照品种郑州5389和36个含有已知抗叶锈病基因的载体品种为材料,苗期接种19个具有鉴别力的叶锈菌生理小种进行基因推导,同时利用12个与抗叶锈病基因紧密连锁的分子标记进行分析。为明确其成株期抗性,分别于2016-2017年和2017-2018年在河北保定对小麦农家品种、感病对照品种郑州5389与慢锈品种SAAR进行田间接种,调查并记录田间严重度及普遍率。基因推导和分子标记检测结果显示,在15个小麦农家品种中共检测到7个抗叶锈病基因,其中部分品种还有多个抗性基因,如红狗豆含有Lr1和Lr46;黄花麦含有Lr13和Lr34;大白麦含有Lr14b和Lr26;洋麦含有Lr37和Lr46;成都光头含有Lr34和Lr46;墨脱麦和西山扁穗含有Lr26和Lr46。部分品种含有1个成株期慢叶锈病抗性基因,如同家坝小麦、武都白茧儿、边巴春麦-6、白花麦含有Lr34;红抢麦、白扁穗和白火麦含有Lr46。这些携带有效抗叶锈病基因的农家品种,可为小麦抗叶锈病育种提供抗源。     

Effect of gene Lr34 in the enhancement of resistance to leaf rust of wheat

Summary Leaf rust resistance gene Lr34 is present in many wheat cultivars throughout the world that have shown durable resistance to leaf rust. Fourteen pair-wise combinations of Lr34 and seedling leaf rust resistance genes were developed by intercrossing near isogenic Thatcher lines. In both seedling and adult plant tests homozygous paired combinations of specific resistance genes with Lr34 had enhanced resistance relative to either parent to different numbers of isolates that were avirulent to the additional resistance genes. The TcLr34, 18 line also expressed enhanced resistance to specific isolates virulent to Lr18 in seedling and adult plant stages. In rust nursery tests, homozygous lines were more resistant than either parent, if the additional leaf rust gene conditioned an effective of resistance when present singly. The ability of Lr34 to interact with other genes conditioning effective resistance may contribute to the durability of leaf rust resistance in cultivars with Lr34. Contribution 1453 Agriculture Canada     

Molecular markers for leaf rust resistance genes in wheat

Over 100 genes of resistance to rust fungi: Puccinia recondita f. sp. tritici, (47 Lr - leaf rust genes), P. striiformis (18 Yr - yellow rust genes) and P. graminis f. sp. tritici (41 Sr - stripe rust genes) have been identified in wheat (Triticum aestivum L.) and its wild relatives according to recent papers. Sixteen Lr resistance genes have been mapped using restriction fragments length polymorphism (RFLP) markers on wheat chromosomes. More than ten Lr genes can be identified in breeding materials by sequence tagged site (STS) specific markers. Gene Lrk 10, closely linked to gene Lr 10, has been cloned and its function recognized. Available markers are presented in this review. The STS, cleaved amplified polymorphic sequence (CAPS) and sequence characterized amplified regions (SCAR) markers found in the literature should be verified using Triticum spp. with different genetic background. Simple sequence repeats (SSR) markers for Lr resistance genes are now also available.     

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.     

新疆的小麦品种（系）苗期和成株期抗叶锈性鉴定

对来自新疆的104个小麦品种、高代品系及35个含有已知抗叶锈基因载体品种,在苗期接种12个中国小麦叶锈菌生理小种进行抗叶锈基因推导分析和分子检测;2007-2008年和2008-2009年连续2年度对这些材料进行成株抗叶锈性鉴定并筛选慢叶锈性品种。研究结果显示,在41个品种中共鉴定出6个已知抗叶锈基因Lr26、Lr34、Lr50、Lr3ka、Lr1和Lr14a,其中Lr26存在于21个品种中,Lr34在17个品种被发现,Lr1和Lr14a分别存在于3个品种中,还有2个品种携带Lr3ka以及1个品种携带Lr50。2年田间抗叶锈性鉴定筛选出7个慢叶锈性品种,可用于小麦抗病育种。     

Homoeologous set of NBS-LRR genes located at leaf and stripe rust resistance loci on short arms of chromosome 1 of wheat

Homoeologous group 1 chromosomes of wheat contain important genes that confer resistance to leaf, stem and stripe rusts, powdery mildew and Russian wheat aphid. A disease resistance gene analog encoding nucleotide binding site-leucine rich repeat (NBS-LRR), designated RgaYr10, was previously identified at the stripe rust resistant locus, Yr10, located on chromosome 1BS distal to the storage protein, Gli-B1 locus. RgaYr10 identified gene members in the homoeologous region of chromosome 1DS cosegregating with the leaf rust resistance gene, Lr21, which originally was transferred from a diploid D genome progenitor. Four RgaYr10 gene members were isolated from chromosome 1DS and compared to two gene members previously isolated from the chromosome 1BS homeologue. NBS-LRR genes tightly linked to stripe rust resistance gene Yr10 on chromosome 1BS were closely related in sequence and structure to NBS-LRR genes tightly linked to leaf rust resistance gene Lr21 located within the homoeologous region on chromosome 1DS. The level of sequence homology was similar between NBS-LRR genes that were isolated from different genomes as compared to genes from the same genome. Electronic Publication     

Genetic mapping of adult plant leaf rust resistance genes Lr48 and Lr49 in common wheat

Hypersensitive adult plant resistance genes Lr48 and Lr49 were named based on their genetic independence of the known adult plant resistance genes. This study was planned to determine genomic locations of these genes. Recombinant inbred line populations derived from crosses involving CSP44 and VL404, sources of Lr48 and Lr49, respectively, and the susceptible parent WL711, were used to determine the genomic locations of these genes. Bulked segregant analyses were performed using multiplex-ready PCR technology. Lr48 in genotype CSP44 was mapped on chromosome arm 2BS flanked by marker loci Xgwm429b (6.1 cM) and Xbarc7 (7.3 cM) distally and proximally, respectively. Leaf rust resistance gene Lr13, carried by the alternate parent WL711, was proximal to Lr48 and was flanked by Xksm58 (5.1 cM) and Xstm773-2 (8.7 cM). Lr49 was flanked by Xbarc163 (8.1 cM) and Xwmc349 (10.1 cM) on chromosome arm 4BL. The likely presence of the durable leaf rust resistance gene Lr34 in both CSP44 and VL404 was confirmed using the tightly linked marker csLV34. Near-isogenic lines for Lr48 and Lr49 were developed in cultivar Lal Bahadur. Genotypes combining Lr13 and/or Lr34 with Lr48 or Lr49 were identified as potential donor sources for cultivar development programs.     

Molecular markers for wheat leaf rust resistance gene Lr41

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