Stripe Rust Resistance Gene Research Articles

Stripe rust resistance gene(s) postulation in wheat germplasm with the help of differentials and tagged molecular markers

Thirteen known Yr gene-associated markers pertaining to genes (Yr5,Yr10,Yr15,Yr24/Yr26) were used to identify the genes in selected wheat germplasm which were found resistant under field conditions at two locations in Punjab, India against stripe rust. In field evaluation, 38 genotypes exhibited highly resistant response, with a final rust severity (FRS) ranging from 0 to TR. Seven genotypes expressed a resistant to moderately resistant response with FRS ranging from 5MR–10S. In race-specific phenotying against most prevalent pathotypes of Pucciniastriiformistritici (46S119,110S119 &238S119) by seedling reaction test (SRT) 14 genotypes (29.2%) were found to be immune (IT = 0), 28 genotypes (58.3%) were resistant (IT = 1), and 3 genotypes (6.3%) were moderately resistant (IT = 2). Yr5 was detected in sixteen lines with the help of two markers Xwmc175 and Xgwm120 linked with Yr5. Yr10 was detected in ten lines with the marker Xpsp3000 and Yr15 was detected in fourteen lines with two linked markers; Xgwm413 and Xgwm273. Likewise, Yr24/26 was detected in 15 lines with two linked markers, namely Xbarc181 and Xbarc187. Based on the race specific phenotyping data and marker data, fourteen lines were found to carry a single gene, 16 showed the presence of two gene combinations, and seven genotypes were found to have a combination of three genes. Frequencies of Yr5, Yr15 and Yr26/Yr24 was high among test wheat germplasm in comparison to Yr10.

Exploring the Genetic Variation of Stripe Rust Foliar and Head Infection in Egyptian Wheat as an Effect of Climate Change

Wheat stripe rust (Puccinia striiformis Westend f. sp. tritici; Pst) is a severe disease that affects wheat-growing areas worldwide. In the last few years, Pst head infection (PstHI) was recorded to cause severe yield damage in Egypt. Little information is known about PstHI. In the current study, a set of 34 Egyptian cultivars were evaluated for their Pst foliar infection (PstFI) and PstHI in two growing seasons (2021 and 2022) in Egypt. High genotypic variations in PstFI and PstHI were found. A very weak correlation was found between the resistance to PstFI and PstHI in both growing seasons. Therefore, the resistance to each symptom might be controlled by different genetic systems. The effect of PstFI and PstHI on kernel traits was investigated. PstHI was found to significantly reduce kernel width and weight in heavily diseased years. Therefore, a significant reduction in wheat yield and quality is expected in case of severe PstHI. Three Egyptian genotypes were resistant to PstFI and PstHI in both growing seasons. These genotypes could help in improving the resistance to both symptoms. However, due to the low genetic distance among them, looking for additional sources of resistance is recommended. In 2021, higher head infection was found compared with 2022. Highly significant differences in maximum temperature, minimum temperature, maximum dew point, and average dew point were found between the two growing seasons. Out of these four factors, dew point and ambient temperature were found to be possible reasons for increasing PstHI. Furthermore, 51 isolines carrying different resistance genes were included in the evaluation. At least ten stripe rust resistance genes were found to be effective against PstFI and PstHI. These ten genes should be included in future breeding programs to improve Pst resistance and understand the effect of climate change on Pst.

Molecular characterization and validation of adult-plant stripe rust resistance gene Yr86 in Chinese wheat cultivar Zhongmai 895.

Adult-plant stripe rust resistance gene Yr86 in Chinese wheat cultivar Zhongmai 895 was mapped to the physical interval 710.2-713.2Mb on the long arm of chromosome 2A. Adult-plant resistance to stripe rust is generally more durable than all-stage resistance. Chinese wheat cultivar Zhongmai 895 showed stable stripe rust resistance at the adult-plant stage. To map the genetic loci underlying its resistance, 171 doubled haploid (DH) lines from a Yangmai 16/Zhongmai 895 cross were genotyped with the wheat 660K SNP chip. Disease severities of the DH population and parents were assessed in four environments. A major QTL designated QYryz.caas-2AL was mapped to interval 703.7-715.3Mb on the long arm of chromosome 2A using both chip-based and KASP (kompetitive allele-specific PCR) marker-based methods, explaining 31.5 to 54.1% of the phenotypic variances. The QTL was further validated in an F2 population of cross Emai 580/Zhongmai 895 with 459 plants and a panel of 240 wheat cultivars using KASP markers. Three reliable KASP markers predicted a low frequency (7.2-10.5%) of QYryz.caas-2AL in the test panel and remapped the gene to the physical interval 710.2-713.2Mb. Based on different physical positions or genetic effects from known genes or QTL on chromosome arm 2AL, the gene was predicted to be a new one for adult-plant stripe rust resistance and was named Yr86. Twenty KASP markers linked to Yr86 were developed in this study based on wheat 660K SNP array and genome re-sequencing. Three of them are significantly associated with stripe rust resistance in natural population. These markers should be useful for marker-assisted selection and also provide a starting point for fine mapping and map-based cloning of the new resistance gene.

Transfer of the high-temperature adult-plant stripe rust resistance gene Yr62 in four Chinese wheat cultivars.

The online version contains supplementary material available at 10.1007/s11032-023-01393-1.

Comparison of gene expression changes in two wheat varieties with different phenotype to strip rust using RNA-Seq analysis

The fungus Puccinia striiformis f. sp. tritici (Pst) is an important threat to wheat production because it can cause wheat stripe rust. The present study aimed to identify new stripe rust resistance genes and to provide a theoretical and practical basis for breeding wheat varieties with broad spectrum, stable, and durable resistance. Wheat leaves inoculated with wheat stripe rust fungus Chinese yellow rust 34 were collected at different time points for transcriptomic analysis based on the wheat stripe rust susceptible varieties AVOCET S (AVS) and AVSYr15NIL [near-isogenic line (NIL) derived from AVS]. The results showed that the number of upregulated genes in the two varieties was 294, 364, 398, and 604, and the number of downregulated genes was 520, 178, 570, and 345 on the 1st, 3rd, 5th, and 7th days post inoculation, respectively. Gene Ontology and Kyoto Encyclopedia of Gene and Genomes enrichment analyses found enrichment of differentially expressed genes in the peroxisome proliferators-activated receptor signaling pathways, plant–pathogen interaction, and styrene acrylic acid biosynthesis that encoded protein kinases, signal transduction, transcription factors, and functional protein components. Differentially expressed genes were randomly selected for quantitative reverse transcription PCR analysis, and the change trend was the same as in the transcriptome data. The results of this study suggest that genes in AVSYr15NIL related to the stripe rust response could be valuable for understanding the mechanisms involved in stripe rust resistance.

Identification of a suppressor for the wheat stripe rust resistance gene Yr81 in Chinese wheat landrace Dahongpao

Combined with BSE-Seq analysis and multiple genetic populations, three genes involved in stripe rust resistance were identified in Chinese wheat landrace Dahongpao, including a novel suppressor on 2BS. Dahongpao (DHP), a landrace of hexaploid wheat in China, exhibits a high degree of stripe rust resistance in the field for many years. In this study, bulked segregant analysis coupled with exome capture sequencing (BSE-Seq) was used to identify genes encoding stripe rust resistance in multiple genetic populations from the cross between DHP and a susceptible hexaploid Australian cultivar, Avocet S (AvS). The most effective QTL in DHP was Yr18, explaining up to 53.08% of phenotypic variance in the F2:3 families. To identify additional genes, secondary mapping populations SP1 and SP2 were produced by crossing AvS with two resistant lines derived from F2:3 families lacking Yr18. An all-stage resistance gene, Yr.DHP-6AS, was identified via BSE-Seq analysis of SP1. Combined the recombinant plants from both SP1 and SP2, Yr.DHP-6AS was located between KP6A_1.66 and KP6A_8.18, corresponding to the same region as Yr81. In addition, secondary mapping populations SP3 and SP4 were developed by selfing a segregating line from F2:3 families lacking Yr18. A novel suppressor gene on chromosome 2BS was identified from DHP for effectively suppressing the resistance of Yr.DHP-6AS in the SP3 and SP4. As a result, the wheat lines carrying both Yr18 and Yr.DHP-6AS show higher level of stripe rust resistance than DHP, providing an effective and simple combination for developing new wheat cultivars with ASR and APR genes. Further, the newly developed KASP markers, KP6A_1.99 and KP6A_5.22, will facilitate the application of Yr.DHP-6AS in wheat breeding via marker-assisted selection.

Identification of a Small Translocation from 6R Possessing Stripe Rust Resistance to Wheat.

Wheat stripe rust, caused by Puccinia striiformis f. sp. tritici Eriks. & E. Henn, is the most devastating fungal disease of bread wheat. Here, a wheat-rye multiple disomic substitution line, SLU126 4R (4D), 5R (5D), and 6R (7D), possessing resistance against 25 races of P. striiformis f. sp. tritici, was used and crossed with Chinese Spring ph1b to induce homeologous recombination to produce introgressions with a reduced rye chromosome segment. Seedling assays confirmed that the stripe rust resistance from SLU126 was retained over multiple generations. Through genotyping-by-sequencing (GBS) platforms and aligning the putative GBS-single-nucleotide polymorphism (SNPs) to the full-length annotated rye nucleotide-binding leucine-rich repeat (NLR) genes in the parental lines (CS ph1b, SLU126, CSA, and SLU820), we identified the physical position of 26, 13, and 9 NLR genes on chromosomes 6R, 4R, and 5R, respectively. The physical positions of 25 NLR genes on chromosome 6R were identified from 568,460,437 bp to 879,958,268 bp in the 6RL chromosome segment. Based on these NLR positions on the 6RL chromosome segment, the three linked SNPs (868,123,650 to 873,285,112 bp) were validated through kompetitive allele-specific PCR (KASP) assays in SLU126 and resistance plants in the family 29-N3-5. Using these KASP markers, we identified a small piece of the rye translocation (i.e., as a possible 6DS.6DL.6RL.6DL) containing the stripe resistance gene, temporary designated YrSLU, within the 6RL segment. This new stripe rust resistance gene provides an additional asset for wheat improvement to mitigate yield losses caused by stripe rust.

Pyramiding of Adult-Plant Resistance Genes Enhances All-Stage Resistance to Wheat Stripe Rust.

Stripe rust, caused by Puccinia striiformis f. sp. tritici, is one of the most destructive diseases in wheat production. Pyramiding of adult-plant resistance (APR) genes is a promising strategy to increase durability of resistance. The stripe rust resistance (R) genes Yr18, Yr28, and Yr36 encode different protein families which confer partial resistance to a broad array of P. striiformis f. sp. tritici races. Here, we developed BC3F5 wheat lines representing all possible combinations of Yr18, Yr28, and Yr36 in a genetic background of the highly P. striiformis f. sp. tritici-susceptible wheat line SY95-71 that is widely used in stripe rust analysis. These lines enabled us to accurately evaluate these genes singly and in combination in a common genetic background. The adult plant resistance experiments were analyzed in the field, where stripe rust epidemics occurred frequently. The field results indicated that these partial R genes act additively in enhancing the levels of resistance, and a minimum of two-gene combinations can generate adequate stripe rust resistance. The Yr28 + Yr36 and Yr18 + Yr28 + Yr36 combinations also showed adequate resistance at the seedling stage, implying that APR gene pyramiding can achieve all-stage resistance. Meanwhile, the three genes were simultaneously introduced into elite wheat lines through gene-based marker selection. Elite lines exhibited strong all-stage resistance to stripe rust. This work provides valuable insights and resources for developing durable P. striiformis f. sp. tritici-resistant varieties and for elucidating the regulation mechanism of partial R gene pyramiding.

High density mapping of wheat stripe rust resistance gene QYrXN3517-1BL using QTL mapping, BSE-Seq and candidate gene analysis.

Fine mapping of a major stripe rust resistance locus QYrXN3517-1BL to a 336kb region that includes 12 candidate genes. Utilization of genetic resistance is an effective strategy to control stripe rust disease in wheat. Cultivar XINONG-3517 (XN3517) has remained highly resistant to stripe rust since its release in 2008. To understand the genetic architecture of stripe rust resistance, Avocet S (AvS) × XN3517 F6 RIL population was assessed for stripe rust severity in five field environments. The parents and RILs were genotyped by using the GenoBaits Wheat 16K Panel. Four stable QTL from XINONG-3517 were detected on chromosome arms 1BL, 2AL, 2BL, and 6BS, named as QYrXN3517-1BL, QYrXN3517-2AL, QYrXN3517-2BL, and QYrXN3517-6BS, respectively. Based on the Wheat 660K array and bulked segregant exome sequencing (BSE-Seq), the most effective QTL on chromosome 1BL is most likely different from the known adult plant resistance gene Yr29 and was mapped to a 1.7cM region [336kb, including twelve candidate genes in International Wheat Genome Sequencing Consortium (IWGSC) RefSeq version 1.0]. The 6BS QTL was identified as Yr78, and the 2AL QTL was probably same as QYr.caas-2AL or QYrqin.nwafu-2AL. The novel QTL on 2BL was effective in seedling stage against the races used in phenotyping. In addition, allele-specifc quantitative PCR (AQP) marker nwafu.a5 was developed for QYrXN3517-1BL to assist marker-assisted breeding.

Development and cytological characterization of wheat-Thinopyrum intermedium translocation lines with novel stripe rust resistance gene.

Wheat stripe rust is a destructive disease in many cool and temperate regions around the world. Exploiting novel sources of resistance can provide wheat cultivars with robust and durable resistance to stripe rust. The wheat-Thinopyrum intermedium addition line TAI-14 was proven to carry a stripe rust resistance gene (named as YrT14) on the alien Th. intermedium chromosome. In order to transfer the resistance gene to wheat, wheat-Th. intermedium translocation lines were created by irradiating the pollen of the line TAI-14. We totally obtained 153 wheat-Th. intermedium translocation lines, among which the long alien segmental translocation line Zhongke 78 and the intercalary translocation line Zhongke 15 not only showed good integrated agronomic traits but also were identified as highly resistant to stripe rust in both seedling and adult plant stages. The alien chromatin in Zhongke 15 was identified as an insertion into the satellite of chromosome 6B, a type of translocation never reported before in chromosome engineering. By screening Simple Sequence Repeat (SSR) and Expressed Sequence Tag (EST) markers as well as the markers developed from RNA-sequencing (RNA-Seq) data, 14 markers were identified specific for the alien chromosome and a physical map was constructed. Both Zhongke 78 and Zhongke 15 could be used as a novel source of stripe rust resistance for wheat breeding, and the linked marker T14K50 can be used for molecular marker-assisted breeding. Finally, based on the karyotype, reaction to stripe rust, and genome resequencing data of different wheat-Th. intermedium translocation lines, the stripe rust resistance gene YrT14 was located to an 88.1 Mb interval from 636.7 to 724.8 Mb on Th. intermedium chromosome 19 corresponding to 7J or 7Js.

Multivariate analysis of selected agro-morphological traits of common wheat genotypes for estimation of genetic diversity

Estimation of genetic diversity of plant species using agro morphological parameters is of very crucial importance in plant improvement programs. In view of that common wheat (Triticum aestivum L.) genotypes consisting of stripe rust resistant genes were analyzed by cluster analysis, principle component analysis and coefficient of correlation analysis for determination of genetic diversity. Experiment was conducted in fields of Hazara University Mansehra Pakistan. Eight groups were categorized by cluster analysis using UPGMA clustering method. AOC-YR/ATTILA (CGSS97Y0061S-23Y-1B-1B) (Sdf31) and 10790 (sdf2) are reported as more diversified by cluster analysis. Based on principal component analysis the first five clusters showed 88.94% of genetic variation. Grains per spike contributed more (0.7) to total variation in PCI. While Harvest Index was the major contributor (0.8) to total variation in PCII. 1056 (Sdf5), AOC-YR/ATTILA (CGSS97Y0061S-32Y-1B-1B) (sdf30), 1072 (sdf6), 1076 (sdf9), and LALBMONO4*4/PRL//LALB (CGSS99Y00093S-2F1-22Y-9GHB-1GHB) (sdf32) were observed superior in flag leaf area, spike length, grains per spike, plant height, biological yield, plant yield and harvest index. AOC-YR/ATTILA (CGSS97Y0061S-32Y-1B-1B) (Sdf30) was found as early maturing genotype. Correlation analysis showed positive and significant association of plant yield with flag leaf area, spike length and biological yield, while it is highly negatively correlated to no. of days to maturity. More diversified genotypes identified in this studies can be used in future breeding programs along with utilization of superior genotypes in hybridization.
 Key words: cluster analysis, correlation, genetic diversity, principal component analysis, whea

Molecular and Cytogenetic Identification of Wheat-Thinopyrum intermedium Double Substitution Line-Derived Progenies for Stripe Rust Resistance.

Thinopyrum intermedium (2n = 6x = 42, JJJSJSStSt) has been hybridized extensively with common wheat and proven to be a valuable germplasm source for improving disease resistance and yield potential of wheat. A novel disease-resistant wheat-Th. intermedium double substitution line X479, carrying 1St(1B) and 4St-4JS (4B), was identified using multi-color non-denaturing fluorescence in situ hybridization (ND-FISH). With the aim of transferring Thinopyrum-specific chromatin to wheat, a total of 573 plants from F2 and F3 progenies of X479 crossed with wheat cultivar MY11 were developed and characterized using sequential ND-FISH with multiple probes. Fifteen types of wheat-Thinopyrum translocation chromosomes were preferentially transmitted in the progenies, and the homozygous wheat-1St, and wheat-4JSL translocation lines were identified using ND-FISH, Oligo-FISH painting and CENH3 immunostaining. The wheat-4JSL translocation lines exhibited high levels of resistance to stripe rust prevalent races in field screening. The gene for stripe rust resistance was found to be physically located on FL0-0.60 of the 4JSL, using deletion lines and specific DNA markers. The new wheat-Th. intermedium translocation lines can be exploited as useful germplasms for wheat improvement.

Description of U6719‐004 wheat germplasm with YrAS2388R stripe rust resistance introgression from Aegilops tauschii

AbstractIn recent years, stripe rust has become an increasingly problematic disease affecting wheat (Triticum aestivum L.). Although stripe rust resistance genes Yr28 and YrAS2388R have been successfully transferred from Aegilops tauschii Coss. to hexaploid wheat, these genes have not been widely used for stripe rust resistance breeding largely due to a lack of publicly available and well‐adapted germplasm. Here we describe U6719‐004 (Entry 538 of D‐genome nested association mapping population, Reg. No. MP‐14, NSL 536301 MAP), a hexaploid wheat germplasm carrying YrAS2388R stripe rust resistance derived from Ae. tauschii accession TA1718 that is well adapted to U.S. wheat production environments. U6719‐004 (KS05HW14‐3/3/KS05HW14‐3/TA1718//KS05HW14‐3) was developed using a combination of direct hybridization of TA1718 with the hard white winter wheat line KS05HW14‐3 followed by two rounds of backcrossing and three generations of single seed descent. U6719‐004 stripe rust resistance was characterized under controlled and natural conditions. Using gene‐based markers, TA1718 and U6719‐004 were confirmed to be YrAS2388R carriers. Grain yield for U6719‐004 was evaluated in 11 year‐by‐location environments across the United States. Grain quality parameters were measured in four year‐by‐location environments. Compared with the recurrent wheat parent KS05HW14‐3 and locally adapted check cultivars, the Ae. tauschii introgression had little to no impact on U6719‐004 grain yield or quality. The availability of U6719‐004 will enable the use of YrAS2388R for stripe rust resistance breeding efforts.

Molecular and cytogenetic dissection of stripe rust resistance gene Yr83 from rye 6R and generation of resistant germplasm in wheat breeding.

Rye 6R-derived stripe rust resistance gene Yr83 in wheat background was physically mapped to fraction length (FL) 0.87-1.00 on the long arm by non-denaturing-fluorescence in situ hybridization (ND-FISH), Oligo-FISH painting and 6R-specific PCR markers.Stripe rust resistance gene Yr83 derived from chromosome 6R of rye (Secale cereale) “Merced” has displayed high resistance to both Australian and Chinese wheat stripe rust isolates. With the aim to physically map Yr83 to a more precise region, new wheat- 6R deletion and translocation lines were produced from derived progenies of the 6R(6D) substitution line. The non-denaturing fluorescence in situ hybridization (ND-FISH) patterns of 6R were established to precisely characterize the variations of 6R in different wheat backgrounds. Comparative ND-FISH analysis localized the breakpoints of 6RL chromosomes relative to Oligo-pSc200 and Oligo-pSc119.2 rich sites in deletion lines. Molecular marker and resistance analyses confirmed that Yr83 is physically located at the fraction length (FL) 0.87-1.00 of 6RL and covers the corresponding region of 806-881 Mb in the reference genome of Lo7. Oligo-FISH painting demonstrated that the region carrying Yr83 is syntenic to the distal end of long arm of homoeologous group 7 of the Triticeae genome. The developed wheat-6R lines carrying the Yr83 gene will be useful for breeding for rust resistance.

Haplotype variants of the stripe rust resistance gene Yr28 in Aegilops tauschii.

Stripe rust resistance gene YrAet672 from Aegilops tauschii accession CPI110672 encodes a nucleotide-binding and leucine-rich repeat domain containing protein similar to YrAS2388 and both these members were haplotypes of Yr28. New sources of host resistance are required to counter the continued emergence of new pathotypes of the wheat stripe rust pathogen Puccinia striiformis Westend. f. sp. tritici Erikss. (Pst). Here, we show that CPI110672, an Aegilops tauschii accession from Turkmenistan, carries a single Pst resistance gene, YrAet672, that is effective against multiple Pst pathotypes, including the four predominant Pst lineages present in Australia. The YRAet672 locus was fine mapped to the short arm of chromosome 4D, and a nucleotide-binding and leucine-rich repeat gene was identified at the locus. A transgene encoding the YrAet672 genomic sequence, but lacking a copy of a duplicated sequence present in the 3' UTR, was transformed into wheat cultivar Fielder and Avocet S. This transgene conferred a weak resistance response, suggesting that the duplicated 3' UTR region was essential for function. Subsequent analyses demonstrated that YrAet672 is the same as two other Pst resistance genes described in Ae. tauschii, namely YrAS2388 and Yr28. They were identified as haplotypes encoding identical protein sequences but are polymorphic in non-translated regions of the gene. Suppression of resistance conferred by YrAet672 and Yr28 in synthetic hexaploid wheat lines (AABBDD) involving Langdon (AABB) as the tetraploid parent was associated with a reduction in transcript accumulation.

Fine mapping of powdery mildew and stripe rust resistance genes Pm5V/Yr5V transferred from Dasypyrum villosum into wheat without yield penalty.

The novel wheat powdery mildew and stripe rust resistance genes Pm5V/Yr5V are introgressed from Dasypyrum villosum and fine mapped to a narrowed region in 5VS, and their effects on yield-related traits were characterized. The powdery mildew and stripe rust seriously threaten wheat production worldwide. Dasypyrum villosum (2n = 2x = 14, VV), a relative of wheat, is a valuable resource of resistance genes for wheat improvement. Here, we describe a platform for rapid introgression of the resistance genes from D. villosum into the wheat D genome. A complete set of new wheat-D. villosum V (D) disomic substitution lines and 11 D/V Robertsonian translocation lines are developed and characterized by molecular cytogenetic method. A new T5DL·5V#5S line NAU1908 shows resistance to both powdery mildew and stripe rust, and the resistances associated with 5VS are confirmed to be conferred by seedling resistance gene Pm5V and adult-plant resistance gene Yr5V, respectively. We flow-sort chromosome arm 5VS and sequence it using the Illumina NovaSeq 6000 system that allows us to generate 5VS-specific markers for genetic mapping of Pm5V/Yr5V. Fine mapping shows that Pm5V and Yr5V are closely linked and the location is narrowed to an approximately 0.9Mb region referencing the sequence of Chinese Spring 5DS. In this region, a NLR gene in scaffold 24,874 of 5VS orthologous to TraesCS5D02G044300 is the most likely candidate gene for Pm5V. Soft- and hard-grained T5DL·5V#5S introgressions confer resistance to both powdery mildew and stripe rust in diverse wheat genetic backgrounds without yield penalty. Meanwhile, significant decrease in plant height and increase in yield were observed in NIL-5DL·5V#5S compared with that in NIL-5DL·5DS. These results indicate that Pm5V/Yr5V lines might have the potential value to facilitate wheat breeding for disease resistance.

Fine mapping of a stripe rust resistance gene YrZM175 in bread wheat.

A stripe rust resistance gene YrZM175 in Chinese wheat cultivar Zhongmai 175 was mapped to a genomic interval of 636.4 kb on chromosome arm 2AL, and a candidate gene was predicted. Stripe rust, caused by Puccinia striiformis f. sp. tritici (PST), is a worldwide wheat disease that causes large losses in production. Fine mapping and cloning of resistance genes are important for accurate marker-assisted breeding. Here, we report the fine mapping and candidate gene analysis of stripe rust resistance gene YrZM175 in a Chinese wheat cultivar Zhongmai 175. Fifteen F1, 7,325 F2 plants and 117 F2:3 lines derived from cross Avocet S/Zhongmai 175 were inoculated with PST race CYR32 at the seedling stage in a greenhouse, and F2:3 lines were also evaluated for stripe rust reaction in the field using mixed PST races. Bulked segregant RNA-seq (BSR-seq) analyses revealed 13 SNPs in the region 762.50-768.52Mb on chromosome arm 2AL. By genome mining, we identified SNPs and InDels between the parents and contrasting bulks and mapped YrZM175 to a 0.72-cM, 636.4-kb interval spanned by YrZM175-InD1 and YrZM175-InD2 (763,452,916-764,089,317bp) including two putative disease resistance genes based on IWGSC RefSeq v1.0. Collinearity analysis indicated similar target genomic intervals in Chinese Spring, Aegilops tauschii (2D: 647.7-650.5Mb), Triticum urartu (2A: 750.7-752.3Mb), Triticum dicoccoides (2A: 771.0-774.5Mb), Triticum turgidum (2B: 784.7-788.2Mb), and Triticum aestivum cv. Aikang 58 (2A: 776.3-778.9Mb) and Jagger (2A: 789.3-791.7Mb). Through collinearity analysis, sequence alignments of resistant and susceptible parents and gene expression level analysis, we predicted TRITD2Bv1G264480 from Triticum turgidum to be a candidate gene for map-based cloning of YrZM175. A gene-specific marker for TRITD2Bv1G264480 co-segregated with the resistance gene. Molecular marker analysis and stripe rust response data revealed that YrZM175 was different from genes Yr1, Yr17, Yr32, and YrJ22 located on chromosome 2A. Fine mapping of YrZM175 lays a solid foundation for functional gene analysis and marker-assisted selection for improved stripe rust resistance in wheat.

Stripe rust and leaf rust resistance in CIMMYT wheat line “Mucuy” is conferred by combinations of race-specific and adult-plant resistance loci

Developing wheat varieties with durable resistance is a core objective of the International Maize and Wheat Improvement Center (CIMMYT) and many other breeding programs worldwide. The CIMMYT advanced wheat line “Mucuy” displayed high levels of resistance to stripe rust (YR) and leaf rust (LR) in field evaluations in Mexico and several other countries. To determine the genetic basis of YR and LR resistance, 138 F5 recombinant inbred lines (RILs) derived from the cross of Apav#1× Mucuy were phenotyped for YR responses from 2015 to 2020 at field sites in India, Kenya, and Mexico, and LR in Mexico. Seedling phenotyping for YR and LR responses was conducted in the greenhouse in Mexico using the same predominant races as in field trials. Using 12,681 polymorphic molecular markers from the DArT, SNP, and SSR genotyping platforms, we constructed genetic linkage maps and QTL analyses that detected seven YR and four LR resistance loci. Among these, a co-located YR/LR resistance loci was identified as Yr29/Lr46, and a seedling stripe rust resistance gene YrMu was mapped on the 2AS/2NS translocation. This fragment also conferred moderate adult plant resistance (APR) under all Mexican field environments and in one season in Kenya. Field trial phenotyping with Lr37-virulent Puccinia triticina races indicated the presence of an APR QTL accounting for 18.3–25.5% of the LR severity variation, in addition to a novel YR resistance QTL, QYr.cim-3DS, derived from Mucuy. We developed breeder-friendly KASP and indel molecular markers respectively for Yr29/Lr46 and YrMu. The current study validated the presence of known genes and identified new resistance loci, a QTL combination effect, and flanking markers to facilitate accelerated breeding for genetically complex, durable rust resistance.

Marker-assisted transfer of leaf and stripe rust resistance from Triticum turgidum var. durum cv. Trinakria to wheat variety HD2932.

A marker-assisted backcrossing program initiated to transfer leaf rust resistance gene LrTrk from Triticum turgidum cv. Trinakria to hexaploid wheat variety HD2932 cotransferred a stripe rust resistance gene, YrTrk, along with LrTrk. The cross of hexaploid recurrent parent HD2932 with tetraploid donor parent Trinakria produced pentaploid F1 plants. F1s were backcrossed with recurrent parent HD2932 to produce BC1F1 generation. Foreground and background selection was conducted in each backcross generation to identify plants for backcrossing or selfing. While foreground selection for LrTrk was carried out with linked and validated molecular marker Xgwm234, for background selection, 86 polymorphic SSR markers from the A and B genomes were used. Single selected plants from BC1F1 and BC2F1 generations backcrossed and selfed to produce BC2F1and BC2F2 generations, respectively. Background selection resulted in 83.72%, 91.86%, and 98.25% of RPG recovery in BC1F1, BC2F1, and BC2F2 generations, respectively. A total of 27 plants with LrTrk in homozygous state were identified in BC2F2 generation and selfed to produce 27 BC2F3 NILs. All the NILs were tested for leaf and stripe rust resistance at the seedling stage using seven Puccinia triticina and one Puccinia striiformis f.sp. tritici rust pathotypes. All the 27 NILs were found to be resistant to both leaf and stripe rust pathotypes. So, these NILs are designated to carry leaf and stripe rust resistance genes LrTrk/YrTrk.

Establishment and identification of six wheat-Thinopyrum ponticum disomic addition lines derived from partial amphiploid Xiaoyan 7430.

Six wheat-Thinopyrum ponticum disomic addition lines derived from partial amphiploid Xiaoyan 7430 were identified using in situ hybridization and SNP microarray, the homoeologous group and stripe rust resistance of each alien chromosome were determined, and Th. ponticum chromosome-specific markers were developed. Xiaoyan 7430 is a significant partial amphiploid, which is used to set up a bridge for transferring valuable genes from Thinopyrum ponticum (Podp.) Barkworth & D.R. Dewey into common wheat. To accelerate the application of these useful genes in enriching the genetic variability of cultivated wheat by chromosome engineering, a complete set of derived addition lines has been created from Xiaoyan 7430. The chromosome composition of each line was characterized by the combination of genomic in situ hybridization and multicolor fluorescence in situ hybridization (mc-FISH), and the homoeology of each alien chromosome was determined by wheat SNP microarray analysis. Addition line WTA55 with alien group-6 chromosome was evaluated resistant to stripe rust isolates at both the seedling and grain-filling stages (Zadoks scale at z.11 and z.73). Diagnostic marker analysis proved that it could carry a novel stripe rust resistance gene derived from Th. ponticum. Furthermore, a FISH probe and 45 molecular markers specific for alien chromosomes were developed based on specific-locus amplified fragment sequencing (SLAF-seq). Of which 27 markers were separately located on single alien chromosome, and some of them could be used to identify the derived translocation lines. This set of addition lines as well as the molecular markers and the FISH probe will promote the introgression of abundant variation from Th. ponticum into wheat in wheat improvement programs.