Chromosome Arm 1RS Research Articles

Rye-durum wheat 1BL.1RS translocation: implications for drought tolerance and nutritional status.

The translocation of the short chromosome arm 1RS of rye onto the 1B chromosome of common wheat has been shown to improve resistance to stress and yield. Here, translocation was operated in durum wheat and its effects on drought tolerance were evaluated. Both the 1BL.1RS translocation line (Svevo 1BL.1RS) and the corresponding Svevo control were exposed to drought for 7 days. Significant differences were found in root morphology between Svevo and Svevo 1BL.1RS under control and drought conditions. Although Svevo 1BL.1RS experienced more severe growth inhibition due to drought than Svevo, it exhibited greater resilience to oxidative stress. Furthermore, several drought-responsive genes were upregulated in both shoots and roots only in the translocation line. Notably, in roots of Svevo 1BL.1RS, the expression of these genes was also higher in the control condition compared to Svevo, suggesting that these genes could be constitutively expressed at higher levels in the translocation line. Moreover, the 1BL.1RS translocation had a significant impact on the plant's ability to accumulate nutrients under drought. Overall, the impact on sulfate accumulation and the expression of genes associated with its assimilation pathways are particularly noteworthy, highlighting the involvement of sulfur in the plant response to water stress. Additionally, the genetic characterization of Svevo 1BL.1RS revealed variants extending beyond the translocation, located in drought stress-responsive genes.

Molecular Cytogenetic Characterization of Novel Wheat-Rye T1RS.1AL Translocation Lines with Resistance to Powdery Mildew and Stripe Rust Derived from the Chinese Rye Landrace Qinling.

Stripe rust and powdery mildew are serious diseases that severely decrease the yield of wheat. Planting wheat cultivars with powdery mildew and stripe rust resistance genes is the most effective way to control these two diseases. Introducing disease resistance genes from related species into the wheat genome via chromosome translocation is an important way to improve wheat disease resistance. In this study, nine novel T1RS.1AL translocation lines were developed from the cross of wheat cultivar Chuannong25 (CN25) and a Chinese rye Qinling. The results of non-denaturing fluorescence in situ hybridization and PCR showed that all new lines were homozygous for the T1RS.1AL translocation. These new T1RS.1AL translocation lines exhibited strong resistance to stripe rust and powdery mildew. The cytogenetics results indicated that the resistance of the new lines was conferred by the 1RS chromosome arms, which came from Qinling rye. The genetic analysis indicated that there were new dominant resistance genes on the 1RS chromosome arm resistant to stripe rust and powdery mildew, and their resistance patterns were different from those of Yr9, Pm8, and Pm17 genes. In addition, the T1RS.1AL translocation lines generally exhibited better agronomic traits in the field relative to CN25. These T1RS.1AL translocations have great potential in wheat-breeding programs in the future.

Pyramiding of transgenic immune receptors from primary and tertiary wheat gene pools improves powdery mildew resistance in the field.

Introgression of resistance genes from wild or related species is a common strategy to improve disease resistance of wheat cultivars. Pm17 is a gene that confers powdery mildew resistance in wheat. It encodes an NLR type of immune receptor and was introgressed from rye to wheat as part of the 1RS chromosome arm translocation several decades ago. So far it has not been possible to separate Pm17 from its co-introgressed rye genes due to suppressed recombination. Here we tested in the field transgenic Bobwhite wheat overexpressing Pm17 without any other rye genes. Four transgenic events showed high levels of PM17 protein accumulation, strong powdery mildew resistance, and no pleiotropic effects during three field seasons. We used a combined approach of transgene insertion and cross-breeding to generate lines co-expressing Pm17 and Pm3, or Pm17 and Pm8. Blumeria graminis f. sp. tritici infection tests confirmed additive, race-specific resistance of the two pyramided transgenes in lines Pm17+Pm3b and Pm17+Pm8. Furthermore, pyramided lines showed strong powdery mildew resistance during three field seasons. We conclude that the combination of overexpressed NLR genes from the extended gene pool broadens and diversifies wheat disease resistance.

Discovery of Resistance Genes in Rye by Targeted Long-Read Sequencing and Association Genetics

The majority of released rye cultivars are susceptible to leaf rust because of a low level of resistance in the predominant hybrid rye-breeding gene pools Petkus and Carsten. To discover new sources of leaf rust resistance, we phenotyped a diverse panel of inbred lines from the less prevalent Gülzow germplasm using six distinct isolates of Puccinia recondita f. sp. secalis and found that 55 out of 92 lines were resistant to all isolates. By performing a genome-wide association study using 261,406 informative SNP markers, we identified five resistance-associated QTLs on chromosome arms 1RS, 1RL, 2RL, 5RL and 7RS. To identify candidate Puccinia recondita (Pr) resistance genes in these QTLs, we sequenced the rye nucleotide-binding leucine-rich repeat (NLR) intracellular immune receptor complement using a Triticeae NLR bait-library and PacBio® long-read single-molecule high-fidelity (HiFi) sequencing. Trait-genotype correlations across 10 resistant and 10 susceptible lines identified four candidate NLR-encoding Pr genes. One of these physically co-localized with molecular markers delimiting Pr3 on chromosome arm 1RS and the top-most resistance-associated QTL in the panel.

Discovery of a Novel Leaf Rust (Puccinia recondita) Resistance Gene in Rye (Secale cereale L.) Using Association Genomics.

Leaf rust constitutes one of the most important foliar diseases in rye (Secale cereale L.). To discover new sources of resistance, we phenotyped 180 lines belonging to a less well-characterized Gülzow germplasm at three field trial locations in Denmark and Northern Germany in 2018 and 2019. We observed lines with high leaf rust resistance efficacy at all locations in both years. A genome-wide association study using 261,406 informative single-nucleotide polymorphisms revealed two genomic regions associated with resistance on chromosome arms 1RS and 7RS, respectively. The most resistance-associated marker on chromosome arm 1RS physically co-localized with molecular markers delimiting Pr3. In the reference genomes Lo7 and Weining, the genomic region associated with resistance on chromosome arm 7RS contained a large number of nucleotide-binding leucine-rich repeat (NLR) genes. Residing in close proximity to the most resistance-associated marker, we identified a cluster of NLRs exhibiting close protein sequence similarity with the wheat leaf rust Lr1 gene situated on chromosome arm 5DL in wheat, which is syntenic to chromosome arm 7RS in rye. Due to the close proximity to the most resistance-associated marker, our findings suggest that the considered leaf rust R gene, provisionally denoted Pr6, could be a Lr1 ortholog in rye.

Characterization of PmBN418, a wheat powdery mildew resistance gene on the rye 1RS chromosome arm

AbstractPowdery mildew, caused by Blumeria graminis f. sp. tritici (Bgt), is a disease of global importance, and new powdery mildew resistance genes that can be directly used in cultivar development are urgently needed. A recombinant inbred line population from ‘CI 17884’ × ‘Bainong 418’ was genotyped using single nucleotide polymorphism markers generated from genotyping‐by‐sequencing, and evaluated for responses to Great Plains Bgt isolates OKS(14)‐B‐3‐1 and NEI‐1‐3. Genetic mapping identified a powdery mildew resistance gene, designated PmBN418, on a 1BL.1RS translocation segment in Bainong 418. Given that PmBN418 conferred resistance to Bgt isolates virulent to Pm8 that resides on the chromosome arm 1RS translocated from rye (Secale cereale L.) cv. ‘Petkus’, PmBN418 is either a new gene or a new allele of Pm8. Phenotypic and genomic data analyses indicated segregation distortion of the powdery mildew resistance gene due to reduced transmission of the wheat (Triticum aestivum L.)–rye 1BL.1RS translocation segment from Bainong 418, while the wheat–Triticum speltoides translocation segment 7S#1 in CI 17884 was preferentially transmitted. Bainong 418, a high‐yielding cultivar with good resistance to several U.S. Bgt isolates, is a useful resistance source for wheat breeding, and the availability of the Kompetitive Allele Specific PCR (KASP) marker KASP‐1RS‐1 makes marker‐assisted introgression of PmBN418 into locally adapted cultivars feasible.

Characterization of an Incomplete Leaf Rust Resistance Gene on Chromosome 1RS and Development of KASP Markers for Lr47 in Wheat.

Leaf rust, caused by Puccinia triticina, is one of the most common wheat (Triticum aestivum) diseases in the Great Plains of the United States. A population of recombinant inbred lines from CI 17884 × 'Bainong 418' was evaluated for responses to leaf rust race Pt52-2 and genotyped using single nucleotide polymorphism (SNP) markers. Quantitative trait locus analysis identified a minor gene for resistance to leaf rust, designated QLr.stars-1RS, on the 1BL.1RS translocation segment in 'Bainong 418', and another leaf rust resistance gene, Lr47, on chromosome 7A of CI 17884. Lr47, originally identified in CI 17884 and located in a wheat-T. speltoides translocation segment 7S#1S, remains one of only a few race-specific resistance genes still effective in the Great Plains. A set of 7A-specific simple sequence repeat markers were developed and used to genotype CI 17884 and a pair of near-isogenic lines differing in the presence or absence of 7S#1S, PI 603918, and 'Pavon F76'. Haplotype analysis indicated that the estimated length of 7S#1S was 157.23 to 174.42 Megabases, accounting for ∼23% of the 7A chromosome. Two SNPs on 7S#1S and four SNPs on the 1RS chromosome arm were converted to Kompetitive allele-specific PCR (KASP) markers, which were subsequently validated in a panel of cultivars and elite breeding lines released within the last decade. Of these, one- and two-KASP markers are specific to the 1RS chromosome arm and 7S#1S, respectively, indicating that they can facilitate the introgression of Lr47 and QLr.stars-1RS into locally adapted wheat cultivars and breeding lines.

Identification of an Elite Wheat-Rye T1RS·1BL Translocation Line Conferring High Resistance to Powdery Mildew and Stripe Rust.

Wheat-rye T1RS·1BL translocations have been widely used worldwide in wheat production for multiple disease resistance and superior yield traits. However, many T1RS·1BL translocations have successively lost their resistance to pathogens due to the coevolution of pathogen virulence with host resistance. Because of the extensive variation in rye (Secale cereale L.) as a naturally cross-pollinating relative of wheat, it still has promise to widen the variation of 1RS and to fully realize its application value in wheat improvement. In the present study, the wheat-rye breeding line R2207 was characterized by comprehensive analyses using genomic in situ hybridization (GISH), multicolor fluorescence in situ hybridization with multiple probes, multicolor GISH, and molecular marker analysis, and then was proven to be a cytogenetically stable wheat-rye T1RS·1BL translocation line. Based on the disease responses to different isolates of powdery mildew and genetic analysis, R2207 appears to possess a novel variation for resistance, which was confirmed to be located on the rye chromosome arm 1RS. Line R2207 also exhibited high levels of resistance to stripe rust at both seedling and adult stages, as well as enhanced agronomic performance, so it has been transferred into a large number of commercial cultivars using an efficient 1RS-specific kompetitive allele specific PCR marker for marker-assisted selection.

Effects of the 1BL/1RS translocation on 24 traits in a recombinant inbred line population

The 1BL/1RS translocation is widespread in a large number of wheat varieties. In this study, we comprehensively evaluated the effects of the 1BL/1RS translocation on 24 traits in a recombinant inbred line population derived from the cross between 20828 and Chuannong16. The phenotypic data was from multiple environments and major loci for a given trait were excluded to avoid their interference with the effects of 1BL/1RS translocation. Comparison results showed that 1RS chromosome arm carried lines had positive effect on increasing maximum root length and spike density. While 1BS chromosome arm carried lines had positive effects on increasing plant height, grain number per spike, effective tiller, root tip number, and resistance to stripe rust. The stripe rust resistance genes, Yr15 detected on 1BS in this study and QYr.sicau-1B.1 identified previously on 1BL were both derived from 20828. These results revealed the genetic mechanism of near-immune resistance to stripe rust in 20828 that has been utilized as a breeding parent for over a decade, and thus possibly accelerate its breeding utilization. This study combined with previous studies indicated that 1BS chromosome arm is likely beneficial to yield increase, while 1RS is more helpful for drought tolerance improvement.

Persistence of rye (Secale cereale L.) chromosome arm 1RS in wheat (Triticum aestivum L.) breeding programs of the Great Plains of North America

Rye (Secale cereale L.) chromosome arm 1RS has been used world-wide by wheat (Triticum aestivum L.) breeding programs as a source of pest- and pathogen-resistance genes, and to improve grain yield and stress tolerance. The most common vehicles used to access 1RS are various 1AL.1RS and 1BL.1RS wheat-rye chromosomal translocations. Over the past 25 years, advanced North American wheat breeding lines were evaluated, first by assay of secalin storage proteins, and later by use of DNA marker TSM0120, for the presence of these two translocations. Both methods provide accurate and efficient means of identifying and differentiating 1BL.1RS and 1A.1RS. Both 1Al.1RS and 1BL.1RS wheats were found in all tested years. 1AL.1RS lines were more common in southern Great Plains breeding programs. 1AL.1RS lines were released as cultivars at a frequency identical to that of wild-type breeding lines. In contrast, 1BL.1RS breeding lines were developed by breeding programs throughout the Great Plains, but fewer were released as cultivars. Both 1RS translocation types persist in Great Plains breeding programs. The lower rate of release of 1BL.1RS cultivars no doubt is a consequence of the more drastic effects on breadmaking quality relative to those observed with 1AL.1RS.

Evolutionary divergence of the rye Pm17 and Pm8 resistance genes reveals ancient diversity.

We have isolated a novel powdery mildew resistance gene in wheat that was originally introgressed from rye. Further analysis revealed evolutionary divergent history of wheat and rye orthologous resistance genes. Wheat production is under constant threat from a number of fungal pathogens, among them is wheat powdery mildew (Blumeria graminis f. sp. tritici). Deployment of resistance genes is the most economical and sustainable method for mildew control. However, domestication and selective breeding have narrowed genetic diversity of modern wheat germplasm, and breeders have relied on wheat relatives for enriching its gene pool through introgression. Translocations where the 1RS chromosome arm was introgressed from rye to wheat have improved yield and resistance against various pathogens. Here, we isolated the Pm17 mildew resistance gene located on the 1RS introgression in wheat cultivar 'Amigo' and found that it is an allele or a close paralog of the Pm8 gene isolated earlier from 'Petkus' rye. Functional validation using transient and stable transformation confirmed the identity of Pm17. Analysis of Pm17 and Pm8 coding regions revealed an overall identity of 82.9% at the protein level, with the LRR domains being most divergent. Our analysis also showed that the two rye genes are much more diverse compared to the variants encoded by the Pm3 gene in wheat, which is orthologous to Pm17/Pm8 as concluded from highly conserved upstream sequences in all these genes. Thus, the evolutionary history of these orthologous loci differs in the cereal species rye and wheat and demonstrates that orthologous resistance genes can take different routes towards functionally active genes. These findings suggest that the isolation of Pm3/Pm8/Pm17 orthologs from other grass species, additional alleles from the rye germplasm as well as possibly synthetic variants will result in novel resistance genes useful in wheat breeding.

Addition of chromosome 4R from Hungarian rye cultivar Lovászpatonai confers resistance to stripe rust and outstanding end-use quality in wheat

The Hungarian rye (Secale cereale L., 2n = 2x = 14, RR) cultivar Lovászpatonai possesses numerous agronomically useful traits that can be exploited in wheat breeding. This study reports on the production, molecular cytogenetic identification, and evaluation of resistance to wheat stripe (or yellow) rust (Puccinia striiformis f. sp. tritici) under field conditions of a newly developed wheat-rye (Mv9kr1-‘Lovászpatonai’) disomic 4R addition line. Analysis of grain quality properties showed that the transfer of ‘Lovászpatonai’ chromosome 4R into wheat significantly increased the total arabinoxylan and protein content. This genetic material proved to be resistant to stripe rust in 2014 and 2015 when epidemics broke out in Hungary. It is assumed that this addition line carries an effective, new resistance gene different from that on rye chromosome arm 1RS of the 1RS.1BL translocation. The Mv9kr1-‘Lovászpatonai’ 4R addition line is a promising pre-breeding material being the first step towards creating translocation lines in order to transfer stripe rust resistance as well as high protein and dietary fibre content into cultivated wheat without deleterious effects on yield and grain quality.

Engineering the 1BS chromosome arm in wheat to remove the Rf (multi) locus restoring male fertility in cytoplasms of Aegilops kotschyi, Ae. uniaristata and Ae. mutica.

By removing the Rf (multi) locus from chromosome 1BS of wheat via chromosome engineering we were able to generate a resource for the production of male sterile wheats in three new cytoplasms. Cytoplasmic male sterility is an essential component in the development of many hybrid crops. In wheat (Triticum aestivum L.) only the cytoplasm of T. timopheevi cytoplasm has been extensively tested even though many other cytoplasms are also known to produce male sterility. Among them are the cytoplasms of Ae. kotschyi, Ae. uniaristata and Ae. mutica but here male sterility manifests itself only when the 1RS.1BL rye-wheat translocation is present in the nuclear genome. The location of the male fertility restoring gene on the chromosome arm 1BS (Rf (multi) ) has recently been determined using a set of primary recombinants of chromosome arms 1RS with 1BS. Using this knowledge the same recombinants were used to create chromosome arm 1BS in wheat with a small insert from rye that removes the restorer locus. The disomic engineered chromosome 1B1:6 assures male sterility in all three cytoplasms and any standard chromosome 1B in wheat is capable of restoring it. This newly engineered chromosome in combination with the three cytoplasms of Aegilops sp extends the range of possibilities in attempts to create a viable system for hybrid wheat production.

A Mutant with Expression Deletion of Gene Sec-1 in a 1RS.1BL Line and Its Effect on Production Quality of Wheat.

The chromosome arm 1RS of rye (Secale cereal L.) has been used worldwide as a source of genes for agronomic and resistant improvement. However, the 1RS arm in wheat has end-use quality defects that are partially attributable to the presence of ω-secalins, which are encoded by genes at the Sec-1 locus. Various attempts in removing the Sec-1 genes from the 1RS.1BL translocation chromosome have been made. In the present study, two new primary 1RS.1BL translocation lines, T917-26 and T917-15, were developed from a cross between wheat variety “A42912” and Chinese local rye “Weining.” The lines T917-15 and T917-26 carried a pair of intact and homogeneous 1RS.1BL chromosomes. The line T917-26 also harbored an expression deletion of some genes at the Sec-1 locus, which originated from a mutation that occurred simultaneously with wheat-rye chromosome translocations. These results suggest that the accompanying mutations of the evolutionarily significant translocations are remarkable resources for plant improvement. Comparison of translocation lines with its wheat parent showed improvements in the end-use quality parameters, which included protein content (PC), water absorption (WA), sodium dodecyl sulfate sedimentation (SDSS), wet gluten (WG), dry gluten (DG) and dough stickiness (DS), whereas significant reduction in gluten index (GI) and stability time (ST) were observed. These findings indicate that 1RS in wheat has produced a higher amount of protein, although these comprised worse compositions. However, in the T917-26 line that harbored an expression deletion mutation in the Sec-1 genes, the quality parameters were markedly improved relative to its sister line, T917-15, especially for GI and DS (P < 0.05). These results indicated that expression deletion of Sec-1 genes significantly improves the end-use quality of wheat cultivars harboring the 1RS.1BL translocation. Strategies to remove the Sec-1 genes from the 1RS.1BL translocation in wheat improvement are discussed.

1RS translocation in Croatian winter wheat varieties

The translocation between the short arm of chromosome 1R of rye and the long arm of chromosome 1A, 1B or 1D of wheat represents practically the most common introduced foreign genes into the genome of hexaploid wheat. 1RS chromosome arm represents a source of different useful genes, associated with increased disease resistance, improved adaptability, stress tolerance, and increased stability and level of yield. On the other hand, translocation is associated with poor technological quality of wheat as a result of the presence of secalin and reduced number of gluten loci. The aim of this study was to determine the prevalence of 1RS translocation among some Croatian winter wheat varieties using molecular markers. The study included 40 varieties of which 23 Croatian. Four pairs of primers: RIS, SCM9, RYE-NOR and PAW161 were used for determination of translocation. The presence of translocation was determined in 12 of 40 (30%) varieties, while among the Croatian wheat varieties translocation had 8 of 23 (34.78%) varieties (Zlatna Dolina, Barbara, Nova Žitarka, Marija, Prima, Kuna, Koleda and Dea).

Dynamics of Rye Telomeres in a Wheat Background during Early Meiosis

Migration of the telomere of the short arm of rye chromosome 5R (5RS) during bouquet organization is dependent on the conformation that this chromosome adopts in its intact, submetacentric, or truncated, metacentric, form. In order to establish whether the telomere migration dependence on chromosome conformation is a common feature of all rye chromosomes, the behavior of the telomeres of 2 other rye chromosomes, 1R and 6R, with apparent differences in the arm ratio, has been studied at the bouquet stage and compared with that of 5R. The presence of subtelomeric heterochromatic chromomeres in both arms of 1R and 6R, which were visualized by FISH, revealed the position of the adjacent telomeres in the bouquet. While the end of the long arms of both chromosomes was, with some exceptions, always included in the telomere cluster, the end of the short arms failed to migrate to the telomere pole. Disturbed telomere migration was more often observed in the short arm of the submetacentric chromosome 6R than in the short arm of the almost metacentric chromosome 1R. Thus, the chromosomal conformation effect on telomere mobility is a common feature of all rye chromosomes. Incomplete telomere clustering is followed by failure of synapsis and chiasma formation in chromosomes 5R and 6R. Chromosome arm 1RS, which carries the NOR, completes synapsis earlier than 5RS or 6RS, facilitated by the nucleolar fusion that occurs during early zygotene.

Distribution of 1AL.1RS and 1BL.1RS wheat-rye translocations in Triticum aestivum using specific PCR

Chromosome arm 1RS of rye (Secale cereale) is a valuable resource for wheat (Triticum aestivum) improvement. 1AL.1RS and 1BL.1RS translocations play an important role in wheat breeding, since wheat carrying these chromosomal translocations has higher tolerance to biotic and abiotic stress. In this study, the presence of 1RS and the distribution of 1AL.1RS and 1BL.1RS wheat-rye translocations were examined in 66 Iranian cultivars and 70 regional foreign accessions of bread wheat, using three rye-specific primers (“RYER3/F3”, “O-SEC5′-A/O-SEC3′-R”, “PAWS5/S6”). Based on “RyeR3/F3”, the presence of 1RS was verified in 15 (23%) Iranian cultivars and in two (3%) foreign accessions. Further, “O-SEC5′-A/O-SEC3′-R” and “PAWS5/S6” were used to distinguish 1AL.1RS and 1BL.1RS translocations. According to results from these primers, 1BL.1RS was identified in 14 (21%) Iranian cultivars and two (3%) foreign accessions. The results confirm that “Sholeh” is the only cultivar (1.5%), among all cultivars and accessions, that carries 1AL.1RS. This study provides a useful tool in marker-assisted selection of materials containing 1RS, and in the creation of new Iranian common wheat cultivars with a larger genetic diversity in wheat breeding programs.

Diversity resistance to Puccinia striiformis f. sp Tritici in rye chromosome arm 1RS expressed in wheat.

The 1BL.1RS wheat-rye translocation contained in the Russian cultivar Aurora has been the most widespread alien translocation in wheat-breeding programs all over the world. However, following the prevalence of new biotypes of the pathogens, disease-resistance genes in this translocation chromosome have been overcome and consequently they have been eliminated in modern wheat-breeding programs. In this paper, we report on 12 new primary 1BL.1RS translocation lines derived from the crosses of a Chinese high yield wheat cv. Mianyang 11 with three rye cultivars collected from China. GISH, C-banding and PCR techniques using the specific primers for 1BS, 1RS and centromeres of wheat and rye were applied to identify the constitution of chromosomes. The results confirmed that all 1BL.1RS chromosomes in the 12 primary translocation lines contained integrated 1RS chromosome arms. In the resistance analysis using five kinds of Pst pathotypes, the 12 primary translocation lines showed diversity resistance to stripe rust, which contained at least five different new genes (alleles), significantly different from the Yr9 gene coming from Russian wheat cultivar Aurora. The results indicated that the chromosome arm 1RS in the rye population carries abundant yet untapped genes (alleles) for resistance to wheat stripe rust, which would originate from the neutral diversity in the natural population of rye. It is suggested that creating more primary translocation lines in genome modification will be extremely important to use the diversity of alien R-genes, which was generated by long-term neutral mutation and maintained in the population of alien species.

Rye Pm8 and wheat Pm3 are orthologous genes and show evolutionary conservation of resistance function against powdery mildew

The improvement of wheat through breeding has relied strongly on the use of genetic material from related wild and domesticated grass species. The 1RS chromosome arm from rye was introgressed into wheat and crossed into many wheat lines, as it improves yield and fungal disease resistance. Pm8 is a powdery mildew resistance gene on 1RS which, after widespread agricultural cultivation, is now widely overcome by adapted mildew races. Here we show by homology-based cloning and subsequent physical and genetic mapping that Pm8 is the rye orthologue of the Pm3 allelic series of mildew resistance genes in wheat. The cloned gene was functionally validated as Pm8 by transient, single-cell expression analysis and stable transformation. Sequence analysis revealed a complex mosaic of ancient haplotypes among Pm3- and Pm8-like genes from different members of the Triticeae. These results show that the two genes have evolved independently after the divergence of the species 7.5 million years ago and kept their function in mildew resistance. During this long time span the co-evolving pathogens have not overcome these genes, which is in strong contrast to the breakdown of Pm8 resistance since its introduction into commercial wheat 70 years ago. Sequence comparison revealed that evolutionary pressure acted on the same subdomains and sequence features of the two orthologous genes. This suggests that they recognize directly or indirectly the same pathogen effectors that have been conserved in the powdery mildews of wheat and rye.

Resistance to multiple cereal aphids in wheat–alien substitution and translocation lines

Rhopalosiphum padi, Schizaphis graminum, and Sitobion avenae are three of the most destructive aphid species of wheat (Triticum aestivum L.). They can significantly reduce wheat yields directly by feeding and indirectly by transmitting viruses. This study aimed to search for resistance to these aphid species among lines derived from different rye (Secale cereale) origins and from Aegilops speltoides, all in the genetic background of the wheat cultivar Pavon F76. Resistance was quantified as aphid weight (R. padi, S. avenae, and S. graminum) and the number of aphids and percentage of infested leaf area exhibiting chlorosis (S. graminum). The most resistant genotypes reduced R. padi and S. avenae weight by 24.2 and 34.3 %, respectively, at the seedling stage, compared with Pavon F76 control plants. Strong S. graminum resistance was found only in A. speltoides-derived lines, the most resistant of which (7A.7S-L5) sustained just 3 % chlorosis and reduced S. graminum colony weight by 67.7 %. One line carrying the 1AL.1RSam wheat–rye translocation from Amigo wheat (originally from Insave rye) reduced S. avenae weight by 23.2 and 21.8 % in seedling and adult plants, respectively. Single genotypes carrying the complete 1R chromosome or the 1RS chromosome arm derived from E12165 wheat and Presto triticale proved to be resistant to both R. padi and S. avenae at the seedling stage. Further research should be conducted to unravel the genetic basis of resistance to these aphids in 1RS genotypes. The sources of resistance identified here may be useful for incorporating multiple aphid species resistance in wheat breeding programs, particularly for R. padi and S. avenae, to which no resistant wheats have been bred.