The Coiled-Coil NLR <i>Rph1</i>, Confers Leaf Rust Resistance in Barley Cultivar Sudan

Dracatos, Peter M.; Bartoš, Jan; Elmansour, Huda; Singh, Davinder; Karafiátová, Miroslava; Zhang, Peng; Steuernagel, Burkhard; Svačina, Radim; Cobbin, Joanna C. A.; Clark, Bethany; Hoxha, Sami; Khatkar, Mehar S.; Doležel, Jaroslav; Wulff, Brande B. H.; Park, Robert

doi:10.1104/pp.18.01052

Cited by 51 publications

(46 citation statements)

References 63 publications

(76 reference statements)

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“…However, none of the BOPA or GBS SNP markers from this work were found within 100 Kb of the gene reported by Dracatos et al. ().…”

Section: Resultscontrasting

confidence: 71%

“…No other introgressions were detected in BW682 (Supplemental Figure S1), and it was selected as the Rph1 source for the differential host set. Dracatos et al (2019) cloned the candidate gene (HORVU2Hr1G006480.6) for Rph1 at ∼13.5 Mb in chromosome 2HS from Sudan. However, none of the BOPA or GBS SNP markers from this work were found within 100 Kb of the gene reported by Dracatos et al (2019).…”

Section: Crop Sciencementioning

confidence: 99%

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Development of barley introgression lines carrying the leaf rust resistance genes Rph1 to Rph15

et al. 2020

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In many production areas, barley (Hordeum vulgare L.) is attacked by the leaf rust pathogen (Puccinia hordei Otth), a basidiomycetous fungus that reduces both its yield and quality. Many leaf rust resistance genes, known as reaction to P. hordei (Rph) genes, have been described in barley. To differentiate genetic variants for virulence in pathogen populations, plant pathologists use differentials (i.e., sets of host lines carrying different resistance genes). The sources of Rph1–15 were derived from cultivars, landraces, and wild barley (H. vulgare ssp. spontaneum K. Koch) accessions with diverse geographic origins and agromorphological traits. Ideal differential sets comprise single‐gene lines backcrossed to a single adapted accession that is susceptible to all known races of a pathogen. In this study, sources of Rph1–15 and other Rph gene donors were backcrossed to the susceptible barley cultivar Bowman and then genotyped to characterize the chromosomal positions and sizes of introgressions. Overall, 95 Bowman introgression lines for leaf rust resistance were developed and characterized for their rust phenotypes and genotypes. A single line was selected to represent each of the 15 Rph genes for use as the new barley leaf rust differential set. The existence of possible new resistance genes in the studied germplasm was postulated. The new Bowman Rph1–15 differential lines will facilitate the efficient virulence phenotyping of P. hordei and serve as valuable genetic stocks for Rph gene stacking and cloning in barley.

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“…However, none of the BOPA or GBS SNP markers from this work were found within 100 Kb of the gene reported by Dracatos et al. ().…”

Section: Resultscontrasting

confidence: 71%

Section: Crop Sciencementioning

confidence: 99%

Development of barley introgression lines carrying the leaf rust resistance genes Rph1 to Rph15

et al. 2020

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“…Phylogenetic analysis using numerous recently cloned NLR immune receptors from the Triticeae not surprisingly determined Rph15 was most closely related to other BED-domain carrying NLR proteins including both Yr5 and Yr7 homologues from wheat and Xa1 from rice on chromosome 4 (corresponding to 2H in barley) (Supplementary Figure S5). Interestingly, Rph15 clustered with previously cloned NLRs from bread wheat conferring both rust and mildew resistance relative to the MLA clade and the recently cloned leaf rust resistance NLR Rph1 from cultivated barley (Dracatos et al 2019; Supplementary Figure S5).…”

Section: Resultsmentioning

confidence: 86%

“…The presence of NLR clusters and multiple disease resistance alleles with different pathogen specificities arising through duplication and co-evolution is well established in the plant kingdom (Michelmore and Meyers 1998; Dracatos et al 2019). Three leaf rust resistance loci, (i.e.…”

Section: Resultsmentioning

confidence: 99%

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Ancient BED-domain-containing immune receptor from wild barley confers widely effective resistance to leaf rust

Chen

Clark

Martin

et al. 2020

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Leaf rust, caused by Puccinia hordei is a devastating fungal disease affecting barley (Hordeum vulgare subsp. vulgare) production globally. Race-specific resistance (R) genes have been deployed widely; however, their durability is often compromised due to the rapid emergence of virulent P. hordei races, prompting the search for new sources of broad-spectrum resistance. Here we report on the cloning of Rph15, a broadly effective resistance gene derived from the wild progenitor Hordeum vulgare subsp. spontaneum. We demonstrate using introgression mapping, mutation and complementation that Rph15 encodes a coiled-coil nucleotide-binding leucine-rich repeat (NLR) protein with an integrated Zinc-finger BED (ZF-BED) domain. The allelic variation at the Rph15 locus was assessed using barley exome capture data that traced its origin to the western region of the Fertile Crescent bordering Jordan and Israel. To unravel the genetic relationship of two other leaf rust resistance genes (Rph14 and Rph16) mapped at similar locus on chromosome 2H, we re-sequenced the Rph15 gene from the near-isogenic line for Rph15 (Bowman+Rph15) and the two donor accessions of Rph14 (PI 584760) and Rph16, (PI 405292, Hs 680). Both whole genome and Sanger sequencing confirmed that Hs 680 carried Rph15, while Rph14 in PI 584760 was an independent locus. A perfect diagnostic KASP marker was developed and validated to permit efficient introduction of Rph15 into cultivated barley.

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Chromosome analysis and sorting

et al. 2021

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Flow cytometric analysis and sorting of plant mitotic chromosomes has been mastered by only a few laboratories worldwide. Yet, it has been contributing significantly to progress in plant genetics, including the production of genome assemblies and the cloning of important genes. The dissection of complex genomes by flow sorting into the individual chromosomes that represent small parts of the genome reduces DNA sample complexity and streamlines projects relying on molecular and genomic techniques. Whereas flow cytometric analysis, that is, chromosome classification according to fluorescence and light scatter properties, is an integral part of any chromosome sorting project, it has rarely been used on its own due to lower resolution and sensitivity as compared to other cytogenetic methods. To perform chromosome analysis and sorting, commercially available electrostatic droplet sorters are suitable. However, in order to resolve and purify chromosomes of interest the instrument must offer high resolution of optical signals as well as stability during long runs. The challenge is thus not the instrumentation, but the adequate sample preparation. The sample must be a suspension of intact mitotic metaphase chromosomes and the protocol, which includes the induction of cell cycle synchrony, accumulation of dividing cells at metaphase, and release of undamaged chromosomes, is time consuming and laborious and needs to be performed very carefully. Moreover, in addition to fluorescent staining chromosomal DNA, the protocol may include specific labelling of DNA repeats to facilitate discrimination of particular chromosomes. This review introduces the applications of chromosome sorting in plants, and discusses in detail sample preparation, chromosome analysis and sorting to achieve the highest purity in flowsorted fractions, and their suitability for downstream applications.

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The Coiled-Coil NLR Rph1, Confers Leaf Rust Resistance in Barley Cultivar Sudan

Cited by 51 publications

References 63 publications

Development of barley introgression lines carrying the leaf rust resistance genes Rph1 to Rph15

Development of barley introgression lines carrying the leaf rust resistance genes Rph1 to Rph15

Ancient BED-domain-containing immune receptor from wild barley confers widely effective resistance to leaf rust

Chromosome analysis and sorting

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