Adult Plant Resistance Gene Research Articles

Isolate Specificity and Polygenic Inheritance of Resistance in Barley to Diverse Heterologous Puccinia striiformis Isolates.

Barley is a host to Puccinia striiformis f. sp. hordei, and is an intermediate or near nonhost to the formae speciales adapted to wheat (P. striiformis f. sp. tritici) and to barley grass (P. striiformis f. sp. pseudo-hordei). The genetic basis of resistance to these forms of P. striiformis is not well understood. Accordingly, a recombinant inbred line (RIL) population was developed using a P. striiformis-susceptible accession (Biosaline-19) and the immune cultivar Pompadour. We investigated the genetic basis of resistance to four diverse P. striiformis isolates (P. striiformis f. sp. pseudo-hordei, and P. striiformis f. sp. tritici pathotypes 104 E137 A-, 134 E16 A+, and 64 E0 A-). and determined that the immunity in Pompadour at the seedling stage to the different P. striiformis isolates was due to quantitative trait loci (QTL) on chromosomes 1H, 3H, 5H, and 7H with both overlapping and distinct specificities. Further histological analysis confirmed the presence of isolate specificity. The RILs were also assessed in the field for resistance to P. striiformis f. sp. pseudo-hordei, P. striiformis f. sp. hordei, and the leaf rust pathogen (P. hordei) to identify pleiotropic QTL loci effective at the adult plant stage and determine whether the leaf rust resistance in Pompadour (Rph20) was also effective to P. striiformis. RILs that were seedling susceptible to P. striiformis f. sp. pseudo-hordei were resistant in the field, implicating the involvement of adult plant resistance (APR). Additional QTLs were identified on chromosome 7H at the same genetic position as Rph23 (APR to leaf rust), suggesting either pleiotropic resistance or the presence of a stripe rust resistance gene closely linked to or allelic with Rph23. Unlike many pleiotropic APR genes identified and isolated in wheat, our data suggest that the Rph20 locus does not confer resistance to the P. striiformis isolates used in this study (P. striiformis f. sp. hordei [χ2 (independence) = 2.47 P > 0.12] and P. striiformis f. sp. pseudo-hordei [χ2 (independence) = 0.42 P > 0.60]).

Characterization of Leaf Rust and Stripe Rust Resistance in Spring Wheat 'Chilero'.

Since 1984, the 'Chilero' spring wheat line developed by CIMMYT has proven to be highly resistant to leaf rust and stripe rust. Amid efforts to understand the basis of resistance of this line, a recombinant inbred line (RIL) population derived from a cross between Avocet and Chilero was studied. The parents and RILs were characterized in field trials for leaf rust and stripe rust in three locations in Mexico between 2012 and 2015 and genotyped with DArT-array, DArT-GBS, and SSR markers. A total of 6,168 polymorphic markers were used to construct genetic linkage maps. Inclusive composite interval mapping detected four colocated resistance loci to both rust diseases and two stripe rust resistant loci in the Avocet × Chilero population. Among these, the quantitative trait locus (QTL) on chromosome 1BL was identified as a pleotropic adult plant resistance gene Lr46/Yr29, whereas QLr.cim-5DS/QYr.cim-5DS was a newly discovered colocated resistance locus to both rust diseases in Chilero. Additionally, one new stripe rust resistance locus on chromosome 7BL was mapped in the current population. Avocet also contributed two minor colocated resistance QTLs situated on chromosomes 1DL and 4BS. The flanking SNP markers can be converted to breeder friendly Kompetitive Allele Specific PCR (KASP) markers for wheat breeding programs.

Unveilingsources of stripe rust resistance in diverse wheat (Triticum Aestivum L.) germplasm using narrow down methodology: a proof of concept

Stripe rust of wheat caused by the fungal pathogen is a destructive foliar disease of wheat. Thus, it is crucial step to characterize the resistant germplasm for stripe rust in a diverse germplasm pool for their ultimate utilization in efficient crop rust resistance breeding. In the present study, we followed two pronged strategies involving integrated phenotypic and molecular characterization of 440 diverse wheat germplasm lines for rust resistance. The germplasm panel was extensively evaluated in field epiphytotic conditions during two consecutive years. After rigorous screening, 72 accessions were successfully revealed as resistant to moderately resistant to stripe rust. Subsequently, entries were then evaluated for their field agronomicperformances, considering prerequisites for serving as a donor germplasm,particularly for yield and 33 potential rust-resistant accessions were identified. Furthermore, to detect the sources of resistance, accessions were molecular characterized for potential race-specific resistance genes Yr5, Yr10,Yr15, and effective adult plant resistance (APR) gene Lr34/Yr18/pm38. We identified the 22 accessions possessing one or more single resistance genes and two accessions were observed with at least three of them. Moreover, Lr34/Yr18/pm38 was determined to confer resistance when observed along with any of the race-specific genes. Thus, the study not only provides proof of concept methodology to identify candidate resistant sources from large germplasm collections but simultaneouslyconfirmed the contribution of combining race-specific andnon-specific APR genes. The finding could further assist in the potential deployment of resistant genes directly into the stripe rust breeding program by involving marker-assisted approaches.

Rust resistance in faba bean (Vicia faba L.): status and strategies for improvement

Faba bean (Vicia faba L.) is an important grain legume used as food and feed. Its production is threatened by abiotic stresses and diseases, of which rust (Uromyces viciae-fabae) is one of the major diseases in East and North Africa, China and the northern grain growing region of Australia. Understanding the genetic and physiological mechanisms of rust resistance in faba bean is in an early phase. The presence of seedling and adult plant resistance genes has been observed. The resistance most frequently utilised in applied plant breeding is race-specific, where the interaction between resistance genes in the host and avirulence genes in the pathogen confers resistance. The main drawback of using race-specific resistance is lack of durability, when deployed singly. Slow rusting or partial resistance, controlled by multiple genes of small effect, is generally non-race specific, so it can be more durable. We present the current knowledge of host resistance and pathogen diversity and propose rational breeding approaches aided with molecular markers to breed durable rust resistance in faba bean.

Identifying some Additional Rust Resistance Genes in Indian Wheat Varieties Using Robust Markers

A set of forty wheat cultivars comprising bread wheat, durum and triticale identified during 2010–2014 were tested for resistance to Indian pathotypes of leaf, stem and yellow rusts at seedling stage under controlled conditions. Eight Lr genes (Lr1, Lr3, Lr10, Lr13, Lr14a, Lr23, Lr24 and Lr26) were characterized based on differential interactions with specific rust races. Genes Lr23, Lr26 and Lr13 conferred leaf rust resistance in most of the accessions. Three Yr genes (YrA, Yr2 and Yr9) were inferred in 40 genotypes, where Yr2 followed by Yr9 were most frequent in conferring stripe rust resistance. Ten Sr genes, namely, Sr2, Sr5, Sr8a, Sr7b, Sr9b, Sr9e, Sr11, Sr13, Sr24 and Sr31, were postulated in these lines with predominance of Sr11, Sr31 and Sr2. These Lr, Sr and Yr genes were observed singly or in combination. Robust DNA markers were used to identify adult plant resistance genes Yr18/Lr34/Sr57, Lr68 and Sr2 and all stage resistance genes Lr24/Sr24, Sr28 and Yr9/Lr26/Sr31. STS marker iag95 showed presence of Yr9 in four additional cultivars which were resistant to one or more rusts. Gene Sr28 was identified in seven durum cultivars with the wPt7004 marker. This is first report of Sr28 being present in many Indian wheat cultivars. CsGs-STS marker identified Lr68 in nine cultivars.

Investigating successive Australian barley breeding populations for stable resistance to leaf rust.

Genome-wide association studies of barley breeding populations identified candidate minor genes for pairing with the adult plant resistance gene Rph20 to provide stable leaf rust resistance across environments. Stable resistance to barley leaf rust (BLR, caused by Puccinia hordei) was evaluated across environments in barley breeding populations (BPs). To identify genomic regions that can be combined with Rph20 to improve adult plant resistance (APR), two BPs genotyped with the Diversity Arrays Technology genotyping-by-sequencing platform (DArT-seq) were examined for reaction to BLR at both seedling and adult growth stages in Australian environments. An integrated consensus map comprising both first- and second-generation DArT platforms was used to integrate QTL information across two additional BPs, providing a total of four interrelated BPs and 15 phenotypic data sets. This enabled identification of key loci underpinning BLR resistance. The APR gene Rph20 was the only active resistance region consistently detected across BPs. Of the QTL identified, RphQ27 on chromosome 6HL was considered the best candidate for pairing with Rph20. RphQ27 did not align or share proximity with known genes and was detected in three of the four BPs. The combination of RphQ27 and Rph20 was of low frequency in the breeding material; however, strong resistance responses were observed for the lines carrying this pairing. This suggests that the candidate minor gene RphQ27 can interact additively with Rph20 to provide stable resistance to BLR across diverse environments.

Characterization and genome-wide association mapping of resistance to leaf rust, stem rust and stripe rust in a geographically diverse collection of spring wheat landraces

The challenge posed by rapidly changing wheat rust pathogens, both in virulence and in environmental adaptation, calls for the development and application of new techniques to accelerate the process of breeding for durable resistance. To expand the resistance gene pool available for germplasm improvement, a panel of 159 landraces plus old cultivars was evaluated for seedling and adult plant resistance (APR) to over 35 Australian pathotypes of Puccinia triticina, Puccinia graminis f. sp. tritici, and Puccinia striiformis f. sp. tritici. Known seedling resistance (SR) genes for leaf rust (Lr2a, Lr3a, Lr13, Lr23, Lr16, and Lr20), stem rust (Sr12, Sr13, Sr23, Sr30, and Sr36), and stripe rust (Yr3, Yr4, Yr5, Yr9, Yr10, Yr17, and Yr27) were postulated. The APR genes identified via field assessments and marker analyses included the pleiotropic genes (Lr34/Yr18/Sr57, Lr46/Yr29/Sr58, Lr67/Yr46/Sr55, and Sr2/Lr27/Yr30), Lr68, Lr74, and uncharacterized APR. A genome-wide association analysis using linear mixed models detected 79 single nucleotide polymorphism (SNP) markers significantly associated with rust resistance, which were mapped on chromosomes 1A, 1B, 1D, 2A, 2B, 3A, 3B, 3D, 4A, 5A, 5B, 6A, 6B, 6D, 7A, 7B and 7D. SNPs associated with multiple rust resistances probably indicate the presence of new pleiotropic or closely linked genes. SNPs were mapped on chromosome positions (1AL, 1DS, 2AL, 4AS, 5BS, 6DL, and 7AL) that have not been known to carry APR genes. This study revealed the presence of a range of possibly unidentified effective seedling and APRs among the landraces, which might represent new sources of rust resistance for the ongoing effort to develop improved wheat cultivars.

QTL Analysis and Nested Association Mapping for Adult Plant Resistance to Powdery Mildew in Two Bread Wheat Populations.

CIMMYT wheat (Triticum aestivum L.) lines Francolin#1 and Quaiu#3 displayed effective and stable adult plant resistance (APR) to Chinese Blumeria graminis f. sp. tritici isolates in the field. To elucidate their genetic basis of resistance, two recombinant inbred line (RIL) populations of their crosses with Avocet, the susceptible parent, were phenotyped in Zhengzhou and Shangqiu in the 2014–2015 and 2015–2016 cropping seasons. These populations were also genotyped with SSR (simple sequence repeat markers) and DArT (diversity arrays technology) markers. Two common significant quantitative trait loci (QTL) on wheat chromosomes 1BL and 4BL were detected in both populations by joint and individual inclusive composite interval mapping, explaining 20.3–28.7% and 9.6–15.9% of the phenotypic variance in Avocet × Francolin#1 and 4.8–11.5% and 10.8–18.9% in Avocet × Quaiu#3, respectively. Additional QTL were mapped on chromosomes 1DL and 5BL in Avocet × Francolin#1 and on 2DL and 6BS in Avocet × Quaiu#3. Among these, QPm.heau-1DL is probably a novel APR gene contributing 6.1–8.5% of total phenotypic variance. The QTL on 1BL corresponds to the pleiotropic multi-pathogen resistance gene Yr29/Lr46/Pm39, whereas the QTL on 2DL maps to a similar region where stripe rust resistance gene Yr54 is located. The QTL identified can potentially be used for the improvement of powdery mildew and rust resistance in wheat breeding.

Validation and characterization of a QTL for adult plant resistance to stripe rust on wheat chromosome arm 6BS (Yr78)

Key messageThis study validated one QTL for adult plant resistance to stripe rust, identified donor lines of the resistance allele, and demonstrated that it is different from previously namedYrgenes.The spread of more virulent and aggressive races of Puccinia striiformis f. sp. tritici (Pst, causal pathogen of stripe rust) after the year 2000 has caused substantial yield losses worldwide. To find new sources of resistance, we previously performed a genome-wide association study and identified a strong QTL for adult plant resistance on the short arm of chromosome 6B (QYr.ucw-6B). In this study, we validated QYr.ucw-6B in ten biparental populations, and mapped it 0.6 cM proximal to IWA7257 and 3.9 cM distal to IWA4408. We showed that QYr.ucw-6B is located approximately 15 cM proximal to the all-stage resistance gene Yr35 and that none of the resistant lines carries the previously cloned Yr36 gene. Based on these results, QYr.ucw-6B was assigned the name Yr78. This gene was not effective against Pst at the seedling stage, suggesting that it is an adult plant resistance gene. Yr78 has been effective against Pst races present in field experiments performed in the Western USA between 2011 and 2016. Since this gene is predicted to be present at low frequency in wheat germplasm from this region, it can provide a useful tool to diversify the sources of resistance against this devastating pathogen.

Genetics of Leaf Rust Resistance in the Hard Red Winter Wheat Cultivars Santa Fe and Duster

Leaf rust, caused by Puccinia triticina, is a common and important disease of hard red winter wheat (Triticum aeistivum L.) in the Great Plains of the United States. The hard red winter wheat cultivars ‘Santa Fe’ and ‘Duster’ have had effective leaf rust resistance since their release in 2003 and 2006, respectively. The objective of this research was to determine the identity of the leaf rust resistance genes in these cultivars. Both cultivars were vernalized and crossed with the susceptible spring wheat ‘Thatcher’, F1 plants were backcrossed to Thatcher, and 90 backcross (BC) F1 plants from each cross were grown out to develop BC1F2 families. Seedlings of the Thatcher*2/Santa Fe BC1F2 families segregated for two genes. The seedling resistance was correlated with the presence of the 2AS/2NS translocation from Aegilops ventricosa Tausch, which indicated one of the genes was Lr37. In field plots, the families segregated for a single gene that was independent of the seedling resistance and the 2NS translocation. Seedlings of the Thatcher*2/Duster BC1F2 families segregated for two genes. In field plots, the families segregated for two genes that were independent of the seedling resistance genes. Santa Fe has Lr37 on the 2NS translocation, an additional adult plant resistance gene, and was postulated to have Lr3a. A previous postulation of Lr17a in Santa Fe was determined to be incorrect. Duster was postulated to have Lr3a and Lr11 for seedling resistance and, based on a molecular marker, Lr34 plus another adult plant resistance gene for field resistance. Lines with the additional adult plant resistance gene in both crosses had leaf rust severity of 10 to 30% with a resistance response of small to large uredinia with chlorosis and necrosis. Santa Fe and Duster are valuable sources of leaf rust resistance for the southern Great Plains region.

Mapping genes for resistance to stripe rust in spring wheat landrace PI 480035.

Stripe rust caused by Puccinia striiformis Westend. f. sp. tritici Erikks. is an economically important disease of wheat (Triticum aestivum L.). Hexaploid spring wheat landrace PI 480035 was highly resistant to stripe rust in the field in Washington during 2011 and 2012. The objective of this research was to identify quantitative trait loci (QTL) for stripe rust resistance in PI 480035. A spring wheat, “Avocet Susceptible” (AvS), was crossed with PI 480035 to develop a biparental population of 110 recombinant inbred lines (RIL). The population was evaluated in the field in 2013 and 2014 and seedling reactions were examined against three races (PSTv-14, PSTv-37, and PSTv-40) of the pathogen under controlled conditions. The population was genotyped with genotyping-by-sequencing and microsatellite markers across the whole wheat genome. A major QTL, QYr.wrsggl1-1BS was identified on chromosome 1B. The closest flanking markers were Xgwm273, Xgwm11, and Xbarc187 1.01 cM distal to QYr.wrsggl1-1BS, Xcfd59 0.59 cM proximal and XA365 3.19 cM proximal to QYr.wrsggl1-1BS. Another QTL, QYr.wrsggl1-3B, was identified on 3B, which was significant only for PSTv-40 and was not significant in the field, indicating it confers a race-specific resistance. Comparison with markers associated with previously reported Yr genes on 1B (Yr64, Yr65, and YrH52) indicated that QYr.wrsggl1-1BS is potentially a novel stripe rust resistance gene that can be incorporated into modern breeding materials, along with other all-stage and adult-plant resistance genes to develop cultivars that can provide durable resistance.

Rapid identification of a major effect QTL conferring adult plant resistance to stripe rust in wheat cultivar Yaco“S”

Stripe rust is a devastating disease in common wheat (Triticum aestivum) worldwide. Growing cultivars with adult-plant resistance (APR) is an environmental friendly approach that provides long-term protection to wheat from this disease. Wheat cultivar Yaco“S” showed a high level of APR to stripe rust in the field from 2008 to 2014. The objective of this study was to detect the major quantitative trait loci (QTL) for APR to stripe rust in Yaco“S”. One hundred and eighty-four F2:3 lines were developed from a cross between Yaco“S” and susceptible cultivar Mingxian169. Illumina 90K and 660K single nucleotide polymorphism (SNP) chips were implemented to bulked pools and their parents to identify SNPs associated with the major QTL. A high-density linkage map was constructed using simple sequence repeat (SSR) and SNP markers. Inclusive composite interval mapping detected a major effect QTL Qyryac.nwafu-2BS conferring stable resistance to stripe rust in all tested environments. Qyryac.nwafu-2BS were mapped to a 1.3 cm interval and explained 17.3–51.9% of the phenotypic variation. Compared with stripe rust resistance genes previously mapped to chromosome 2B, Qyryac.nwafu-2BS is likely a new APR gene to stripe rust. Combining SNP iSelect assay and kompetitive allele specific PCR technology, we found that the APR gene could be rapidly and accurately mapped and it is useful for improving stripe rust resistance in wheat breeding.

Identification and characterization of seedling and adult plant resistance to Puccinia hordei in selected African barley germplasm

A collection of 112 African barley accessions were assessed for response to Puccinia hordei in seedling greenhouse tests using 10 pathotypes and in adult plant field tests over three successive field seasons in Australia. One of the 10 pathotypes (viz. 5457P+) used in seedling tests was also used in field tests to allow assessment of the presence of adult plant resistance (APR) in lines that were seedling susceptible to this pathotype. The seedling resistance genes Rph1, Rph2, Rph3, Rph9.am and Rph9.z were postulated in a number of accessions, singly and in various combinations, with Rph2 and Rph9.z being the most common. Twenty-six accessions carried seedling resistance that was either uncharacterized or could not be determined using the 10 P. hordei pathotypes. One accession carried high levels of APR and 11 accessions showed moderate levels of APR, all of which were susceptible to all P. hordei pathotypes at the seedling stage. All barley accessions were genotyped for the presence of marker alleles that are closely linked to the APR genes Rph20 and Rph23 (bPb-0837 and Ebmac0603, respectively). No accession was positive for bPb-0837, suggesting that Rph20 is not frequent in African germplasm. Thirteen accessions were postulated to carry Rph23 based on the presence of the marker allele Ebmac0603 found in Yerong (Rph23), and 10 out of the 11 accessions with moderate APR lacked the bPb-0837 and Ebmac0603 marker alleles, indicating that they likely carry new uncharacterized APR genes. Inheritance studies were performed using populations derived from four of the accessions that carried APR (Clho 9776, Clho 11958, Mecknes Maroc and Sinai) by crossing with the susceptible barley genotype Gus. Chi squared analysis of the phenotypic data from F3 populations suggested that CIho9776 carried a single APR gene and CIho11958, Mecknes Maroc and Sinai each carried two genes for APR to leaf rust.

The Lr34 adult plant rust resistance gene provides seedling resistance in durum wheat without senescence

SummaryThe hexaploid wheat (Triticum aestivum) adult plant resistance gene, Lr34/Yr18/Sr57/Pm38/Ltn1, provides broad‐spectrum resistance to wheat leaf rust (Lr34), stripe rust (Yr18), stem rust (Sr57) and powdery mildew (Pm38) pathogens, and has remained effective in wheat crops for many decades. The partial resistance provided by this gene is only apparent in adult plants and not effective in field‐grown seedlings. Lr34 also causes leaf tip necrosis (Ltn1) in mature adult plant leaves when grown under field conditions. This D genome‐encoded bread wheat gene was transferred to tetraploid durum wheat (T. turgidum) cultivar Stewart by transformation. Transgenic durum lines were produced with elevated gene expression levels when compared with the endogenous hexaploid gene. Unlike nontransgenic hexaploid and durum control lines, these transgenic plants showed robust seedling resistance to pathogens causing wheat leaf rust, stripe rust and powdery mildew disease. The effectiveness of seedling resistance against each pathogen correlated with the level of transgene expression. No evidence of accelerated leaf necrosis or up‐regulation of senescence gene markers was apparent in these seedlings, suggesting senescence is not required for Lr34 resistance, although leaf tip necrosis occurred in mature plant flag leaves. Several abiotic stress‐response genes were up‐regulated in these seedlings in the absence of rust infection as previously observed in adult plant flag leaves of hexaploid wheat. Increasing day length significantly increased Lr34 seedling resistance. These data demonstrate that expression of a highly durable, broad‐spectrum adult plant resistance gene can be modified to provide seedling resistance in durum wheat.

Identification and Mapping of Adult Plant Resistance Loci to Leaf Rust and Stripe Rust in Common Wheat Cultivar Kundan

Leaf rust (LR) and stripe rust (YR) are important diseases of wheat worldwide. We used 148 recombinant inbred lines (RIL) from the cross of Avocet × Kundan for determining and mapping the genetic basis of adult plant resistance (APR) loci. The population was phenotyped LR and YR for three seasons in field trials conducted in Mexico and genotyped with the diversity arrays technology sequencing (DArT-Seq) and simple sequence repeat markers. The final genetic map was constructed using 2,937 polymorphic markers with an average distance of 1.29 centimorgans between markers. Inclusive composite interval mapping identified two co-located APR quantitative trait loci (QTL) for LR and YR, two LR QTL, and three YR QTL. The co-located resistance QTL on chromosome 1BL corresponded to the pleiotropic APR gene Lr46/Yr29. QLr.cim-2BL, QYr.cim-2AL, and QYr.cim-5AS could be identified as new resistance loci in this population. Lr46/Yr29 contributed 49.5 to 65.1 and 49.2 to 66.1% of LR and YR variations, respectively. The additive interaction between detected QTL showed that LR severities for RIL combining four QTL ranged between 5.3 and 25.8%, whereas the lowest YR severities were for RIL carrying QTL on chromosomes 1BL + 2AL + 6AL. The high-density DArT-Seq markers across chromosomes can be used in fine mapping of the targeted loci and development SNP markers.

The Future of Lr34 in Modern, High‐Input Wheat Breeding Programs

Lr34 is an important adult plant resistance gene that is effective against a number of wheat (Triticum aestivum L.) diseases and has proved durable for over 100 yr. However, the energy requirements of Lr34 resistance, combined with the pleiotrophic effect of leaf tip necrosis, can result in reduced yield potential. To evaluate the absence of Lr34 from elite breeding material, a cross was made between similar‐yielding, milling wheat cultivars ‘Conquest’ (Lr34sus) and ‘Bakker Gold’ (Lr34res). Independent lines were carried forward and, at the F4 stage, sister pairs were selected for the different homozygous alleles of the marker csLV34, corresponding to Lr34res or Lr34sus. These lines were further selfed to produce nine stable F8 pairs (Lr34res and Lr34sus) for field trialing and evaluation over 3 yr under a range of fungicide treatments. In the absence of fungicide and with natural infections of stripe rust (Puccinia striiformis f.sp. tritici) and leaf rust (Puccinia triticina), Lr34res lines had lower disease severity, increased total yield, and better grain quality than Lr34sus lines. This pattern was reversed under full fungicide control, as Lr34sus lines displayed higher mean plot yield (2010: +4.3%, p < 0.001; 2012: +4.4%, p < 0.001) and possessed superior grain quality data than the Lr34res lines. The high‐yield, high‐input farming environment and the use of full fungicide control in New Zealand's current wheat cultivar trialing system means that genetic sources of rust resistance such as Lr34 provide little or no competitive advantage.

Rapid Phenotyping Adult Plant Resistance to Stem Rust in Wheat Grown under Controlled Conditions.

Stem rust (SR) or black rust caused by Puccinia graminis f. sp. tritici is one of the most common diseases of wheat (Triticum aestivum L.) crops globally. Among the various control measures, the most efficient and sustainable approach is the deployment of genetically resistant cultivars. Traditionally, wheat breeding programs deployed genetic resistance in cultivars, but unknowingly this is often underpinned by a single seedling resistance gene, which is readily overcome by the pathogen. Nowadays, adult plant resistance (APR) is a widely adopted form of rust resistance because more durable mechanisms often underpin it. However, only a handful of SR APR genes are available, so breeders currently strive to combine seedling and APR genes. Phenotyping adult wheat plants for resistance to SR typically involves evaluation in the field. But establishing a rust nursery can be challenging, and screening is limited to once a year. This slows down research efforts to isolate new APR genes and breeding of genetically resistant cultivars.In this study, we report a protocol for rapid evaluation of adult wheat plants for resistance to stem rust. We demonstrate the technique by evaluating apanel of 16 wheat genotypes consisting of near isogenic lines (NILs) for known Sr genes (i.e., Sr2, Sr33, Sr45, Sr50, Sr55, Sr57, and Sr58) and three landraces carrying uncharacterized APR from the N. I. Vavilov Institute of Plant Genetic Resources (VIR). The method can be completed in just 10weeks and involves two inoculations: first conducted at seedling stage and a second at the adult stage (using the same plants). The technique can detect APR, such as that conferred by APR gene Sr2, along with pseudo-black chaff (the morphological marker). Phenotyping can be conducted throughout the year, and is fast and resource efficient. Further, the phenotyping method can be applied to screen breeding populations or germplasm accessions using local or exotic races of SR.

Genetic analysis and mapping of adult plant resistance loci to leaf rust in durum wheat cultivar Bairds.

New leaf rust adult plant resistance (APR) QTL QLr.cim - 6BL was mapped and confirmed the known pleotropic APR gene Lr46 effect on leaf rust in durum wheat line Bairds. CIMMYT-derived durum wheat line Bairds displays an adequate level of adult plant resistance (APR) to leaf rust in Mexican field environments. A recombinant inbred line (RIL) population developed from a cross of Bairds with susceptible parent Atred#1 was phenotyped for leaf rust response at Ciudad Obregon, Mexico, during 2013, 2014, 2015 and 2016 under artificially created epidemics of Puccinia triticina (Pt) race BBG/BP. The RIL population and its parents were genotyped with the 50K diversity arrays technology (DArT) sequence system and simple sequence repeat (SSR) markers. A genetic map comprising 1150 markers was used to map the resistance loci. Four significant quantitative trait loci (QTLs) were detected on chromosomes 1BL, 2BC (centromere region), 5BL and 6BL. These QTLs, named Lr46, QLr.cim-2BC, QLr.cim-5BL and QLr.cim-6BL, respectively, explained 13.5-60.8%, 9.0-14.3%, 2.8-13.9%, and 11.6-29.4%, respectively, of leaf rust severity variation by the inclusive composite interval mapping method. All of these resistance loci were contributed by the resistant parent Bairds, except for QLr.cim-2BC, which came from susceptible parent Atred#1. Among these, the QTL on chromosome 1BL was the known pleiotropic APR gene Lr46, whereas QLr.cim-6BL, a consistently detected locus, should be a new leaf rust resistance locus in durum wheat. The mean leaf rust severity of RILs carrying all four QTLs ranged from 8.0 to 17.5%, whereas it ranged from 10.9 to 38.5% for three QTLs (Lr46+5BL+6BL) derived from the resistant parent Bairds. Two RILs with four QTLs combinations can be used as sources of complex APR in durum wheat breeding.

Genetic mapping of resistance to Puccinia hordei in three barley doubled-haploid populations

The success of breeding for barley leaf rust (BLR) resistance relies on regular discovery, characterization and mapping of new resistance sources. Greenhouse and field studies revealed that the barley cultivars Baronesse, Patty and RAH1995 carry good levels of adult plant resistance (APR) to BLR. Doubled haploid populations [(Baronesse/Stirling (B/S), Patty/Tallon (P/T) and RAH1995/Baudin (R/B)] were investigated in this study to understand inheritance and map resistance to BLR. The seedlings of two populations (B/S and R/B) segregated for leaf rust response that conformed to a single gene ratio ( $${\text{X}}_{1:1}^{2}$$ = 0.12, P > 0.7 for B/S and $${\text{X}}_{1:1}^{2}$$ = 0.34, P > 0.5 for R/B) whereas seedlings of third population (P/T) segregated for two-gene ratio ( $${\text{X}}_{1:1}^{2}$$ = 0.17, P > 0.6) when tested in greenhouse. It was concluded that the single gene in Baudin and one of the two genes in Tallon is likely Rph12, whereas gene responsible for seedling resistance in Stirling is Rph9.am (allele of Rph12). The second seedling gene in Tallon is uncharacterized. In the field, APR was noted in lines that were susceptible as seedlings. A range of disease responses (CI 5–90) was observed in all three populations. Marker trait association analysis detected three QTLs each in populations B/S (QRph.sun-2H.1, QRph.sun-5H.1 and QRph.sun-6H.1) and R/B (QRph.sun-1H, QRph.sun-2H.2, QRph.sun-3H and QRph.sun-6H.2), and four QTLs in population P/T (QRph.sun-6H.2, QRph.sun-1H.2, QRph.sun-5H.2 and QRph.sun-7H) that significantly contributed to low leaf rust disease coefficients. High frequency of QRph. sun-5H.1, QRph. sun-6H.1, QRph. sun-1H.1, QRph. sun-2H.2, QRph. sun-6H.2, QRph. sun-7H (based on presence of the marker, closely associated to the respective QTLs) was observed in international commercial barley germplasm and hence providing an opportunity for rapid integration into breeding programmes. The identified candidate markers closely linked to these QTLs will assist in selecting and assembling new APR gene combinations; expectantly this will help in achieving good levels of durable resistance for controlling BLR.

Mining Vavilov's Treasure Chest of Wheat Diversity for Adult Plant Resistance to Puccinia triticina.

Leaf rust (LR) caused by Puccinia triticina, is among the most important diseases of wheat (Triticum aestivum L.) crops globally. Deployment of cultivars incorporating genetic resistance, such as adult plant resistance (APR) or all-stage resistance, is considered the most sustainable control method. APR is preferred for durability because it places lower selection pressure on the pathogen and is often polygenic. In the search for new sources of APR, here we explored a diversity panel sourced from the N. I. Vavilov Institute of Plant Genetic Resources. Based on DNA marker screening, 83 of the 300 lines were deemed to carry known APR genes; namely, Lr34, Lr46, and Lr67. Interestingly, lines carrying Lr67 were mostly landraces from India and Pakistan, reconfirming the likely origin of the gene. Rapid phenotypic screening using a method that integrates assessment at both seedling and adult growth stages under accelerated growth conditions (i.e., constant light and controlled temperature) identified 50 lines carrying APR. Levels of APR corresponded well with phenotypes obtained in a field nursery inoculated using the same pathotype (R2 = 0.82). The second year of field testing, using a mixture of pathotypes with additional virulence for race-specific APR genes (Lr13 and Lr37), identified a subset of 13 lines that consistently displayed high levels of APR across years and pathotypes. These lines provide useful sources of resistance for future research. A strategy combining rapid generation advance coupled with phenotyping under controlled conditions could accelerate introgression of these potentially novel alleles into adapted genetic backgrounds.