Pathotypes and NIP1 gene sequences of Finnish Rhynchosporium secalis isolates from barley, couch grass and rye

Rhynchosporium secalis (Oud.) J.J. Davis, causal agent of scald of barley (Hordeum vulgare L.), was isolated from barley, couch grass (Elymus repens L.) and rye (Secale cereale L.). Isolates were used to inoculate seedlings of a differential barley series containing several sources of major gene resistance to the disease. The series included Atlas46, (resistance gene Rrs1) and the isogenic line Atlas, which lacks the gene (rrs1). The necrosis-inducing peptide NIP1 has been suggested to be the product of the avirulence gene AvrRrs1 (NIP1) that with the barley resistance gene Rrs1 determines the incompatibility of the pathogen-host interaction. All R. secalis isolates were virulent only on the susceptible barley cultivars Arve and Chan, irrespective of spore concentration or original host species. There were no indications of redundant virulence among the R. secalis isolates. The NIP1 gene was sequenced from each isolate and there was no correlation with source of the isolate or sequence modification and virulence. Four isolates, from barley and couch grass, were characterised by a basic Type I NIP1sequence. The recorded NIP1 sequence changes are consistent withR. secalis populations not receiving selection through deployment of the Rrs1 gene in commercial barley cultivars in Finland. This revised version was published online in July 2006 with corrections to the Cover Date.     

Mapping of a gene for leaf scald resistance in barley line 'B87/14' and validation of microsatellite and RFLP markers for marker-assisted selection

Seedlings of the barley line ‘B87/14’ were resistant to 22 out of 23 Australian isolates of Rhynchosporium secalis, the causal agent of leaf scald.‘B87/14’‐based populations were developed to determine the location of the resistance locus. Scald resistance segregated as a single dominant trait in BC₁F₂ and BC₁F₃ populations. Bulked segregant analysis identified amplified fragment length polymorphisms (AFLPs) with close linkage to the resistance locus. Fully mapped populations not segregating for scald resistance located these AFLP markers on chromosome 3H, possibly within the complex Rrs1 scald locus. Microsatellite and restriction fragment length polymorphism markers adjacent to the AFLP markers were identified and validated for their linkage to scald resistance in a second segregating population, with the closest marker 2.2 cM from the resistance locus. These markers can be used for selection of the Rrs.B87 scald‐resistance locus, and other genes at the chromosome 3H Rrs1 locus.     

The genetic basis of scald resistance in western Canadian barley cultivars

Summary The genetic basis of resistance to scald (Rhynchosporium secalis) within barley breeding populations is poorly understood. The design of effective genetically based resistance strategies is predicated on knowledge of the identity of the resistance genes carried by potential parents. The resistance exhibited by a broad selection of western Canadian barley lines was investigated by evaluating their reactions to five R. secalis isolates. Results were compared to the resistance exhibited by previously characterized lines. This comparison, combined with pedigree analysis indicated that there are two different resistance genes present inwwestern Canadian cultivars. These genes were shown to be independent through analysis of a segregating population derived from a cross between Falcon and CDC Silky. This evidence, along with observed linkage of the gene in CDC Silky with an allele specific amplicon developed for a Rhynchosporium secalis resistance locus on chromosome 3, provides evidence that the gene in Falcon is the Rh2 gene derived from Atlas, and the gene (s) in CDC Silky is located within the Rh/Rh3/Rh4 cluster and is similar to the Rh gene in Hudson.     

Genes for scald resistance from wild barley (Hordeum vulgare ssp spontaneum) and their linkage to isozyme markers

Summary Accessions of Hordeum vulgare ssp. spontaneum, the wild progenitor of barley, collected in Israel (70), Iran (15) and Turkey (6) were screened for seedling response to four isolates of Rhynchosporium secalis, the pathogen causing leaf scald in barley. Resistance was very common in the collection (77%) particularly among accessions from the more mesic sites (90%). The genetics of this resistance were investigated in fifteen backcross (BC₃) lines that contained an isozyme variant from H.v. ssp. spontaneum in a H.v. ssp. vulgare (cv. Clipper) background and were resistant to scald. Segregation in the BC₃F₂ families conformed with a single dominant resistance gene in 9 of the 15 lines. Scald resistance and the isozyme marker were closely linked in three of the BC₃-lines, loosely linked in four and unlinked in the remaining eight. Scald resistance genes were identified on barley chromosomes 1, 3, 4 and 6. Crosses between several of the scald resistant BC-lines together with the linkage data indicated that at least five genetically independent resistances are available for combining together for deployment in barley. The linkage of scald resistance in several BC₃-lines to the isozyme locus Acp2 is of special interest as this locus is highly polymorphic in wild barley.     

A novel barley scald resistance gene: genetic mapping of the Rrs15 scald resistance gene derived from wild barley, Hordeum vulgare ssp. spontaneum

Most genes for resistance to barley leaf scald map either to the Rrs1 locus on the long arm of chromosome 3H, or the Rrs2 locus on the short arm of chromosome 7H. Other loci containing scald resistance genes have previously been identified using lines derived from wild barley, Hordeum vulgare ssp. spontaneum. A single dominant gene conditioning resistance to scald was identified in a third backcross (BC3F3) line derived from an Israeli accession of wild barley. The resistance gene is linked to three microsatellite markers that map to the long arm of chromosome 7H; the closest of these loci, HVM49, maps 11.5 cM from the resistance gene. As no other scald resistance genes have been mapped to this chromosome arm, it is considered to be a novel scald resistance locus. As the Acp2 isozyme locus is linked to this scald resistance locus, at 17.7 cM, Acp2 is assigned to chromosome 7H. Molecular markers linked to the novel scald resistance gene, designated Rrs15, can be used in breeding for scald resistance.     

Genetic mapping of the barley Rrs14 scald resistance gene with RFLP, isozyme and seed storage protein markers

The scald susceptible barley cultivar ‘Clipper’ and a third‐backcross (BC₃) line homozygous for the Rrs14 scald resistance gene that originally came from Hordeum vulgare ssp. spontaneum were grown in replicated field trials. The level of resistance that Rrs14 confers against field populations of the pathogen Rhynchosporium secalis, the causal agent of scald disease, was evaluated. The Rrs14 BC₃ line exhibited 80% and 88% less leaf damage than ‘Clipper’ in 1995 and 1996, respectively. Given this effectiveness of Rrs14, research was undertaken to identify a linked marker locus suitable for indirect selection of Rrs14. Based on linkage to a set of previously mapped loci, Rrs14 was positioned to barley chromosome 1H between the seed storage protein (hordein) loci Hor1 and Hor2, approximately 1.8 cM from the latter locus. The Hor2 locus is thus an ideal codominant molecular marker for Rrs14. The tight linkage between Rrs14 and Hor2 and the availability of alternative biochemical and molecular techniques for scoring Hor2 genotypes, permits simple indirect selection of Rrs14 in barley scald resistance breeding programmes.     

New molecular markers linked to qualitative and quantitative powdery mildew and scald resistance genes in barley for dry areas

A partial genetic linkage map was constructed on 71 doubled-haploid lines derived from a cross between the barley lines Tadmor and WI2291 with 181 molecular markers. The segregating population was used to detect markers linked to the gene Mlg conferring resistance to powdery mildew (Erysiphe graminis f. sp. hordei) and to genes for quantitative resistance to scald (Rhynchosporium secalis). The gene Mlg on chromosome 4H was flanked by two AFLP markers at a distance of 2.0 and 2.4 cM, respectively. QTLs for resistance to scald were detected on chromosomes 2H and 3H. This association of molecular markers with qualitative and quantitative disease resistance loci represents a valuable starting-point for marker-assisted selection. This revised version was published online in July 2006 with corrections to the Cover Date.     

Field screening of simulated segregating barley populations for resistance to scald

Summary Simulated segregating barley populations were screened for resistance to scald (Rhynchosporium secalis) in the field at commercial seeding rates. A reduction in infection on the susceptible component occurred with increasing proportions of resistant genotypes. Similar trends were seen in space planted experiments but the use of susceptible buffer rows counteracted the effect, enhanced the infection in susceptible plants and greatly improved discrimination between resistant and susceptible. These results have been applied to the routine testing of F₂ populations in the barley breeding programme.     

Genetic analysis of resistance to barley scald (Rhynchosporium secalis) in the Ethiopian line `Abyssinian' (CI668)

A doubled haploid barley (Hordeum vulgare L.) population from a cross between the cultivar `Ingrid' and the Ethiopian landrace `Abyssinian' was mapped by AFLP, RFLP, SSR and STS markers and tested for resistance to isolates`4004', `2', `16-6', `17', `22' and `WRS 1872' of Rhynchosporium secalis (Oudem.) J.J. Davis, the causal agent of leaf scald. Resistance tests were conducted on parents, DH-lines, a near-isogenic line of `Abyssinian' (NIL) into `Ingrid', and an F₂ population descended from the same F₁ plants as the DHs. The DH population segregated for at least two major R. secalis resistance QTL. All isolates tested identified a major QTL on chromosome 3 (3H) associated with R. secalis resistance, in a 4 cM support interval between the co-segregating markers Bmac0209/Falc666 and MWG680. The QTL was linked with the markers Falc666 (2.3 cM), YLM/ylp (0.3 cM), MWG680 (1.7 cM), cttaca2 (2.5 cM) and agtc17 (9.8 cM). The second QTL was located on chromosome 1 (7H).However, this QTL was only detected by one isolate and was located in an interval of 16 cM in the distal part of the chromosome. At this QTL the allele for improved scald resistance originated from the parent `Ingrid'. There were a number of minor QTL on chromosomes 2 (2H), 4 (4H) and 6 (6H) that were not repeatable either across replications or analysis methods. The importance of checking QTL-models by cross-validation is stressed. This revised version was published online in August 2006 with corrections to the Cover Date.     

Pathogenic variation among isolates of Rhynchosporium secalis from cultivated barley growing in Victoria,Australia

Summary Three hundred and nineteen Rhynchosporium secalis isolates from cultivated barley were divided into five groups on the basis of their virulence on 15 differential barley varieties. Pathogenic variation was also demonstrated for isolates from different scald lesions within the same crop and amongst different spores from the same lesions.     

Identification of QTLs for powdery mildew and scald resistance in barley

A population of 103 recombinant inbred lines (RILs, F₉-derived lines) developed from the two-row spring barley cross L94 × ‘Vada’ was evaluated under field conditions for resistance against powdery mildew (Blumeria graminis f.sp. hordei) and scald (Rhynchosporium secalis). Apart from the major resistance gene mlo on chromosome 4 (4H), three QTLs (Rbgq1, Rbgq2 and Rbgq3) for resistance against powdery mildew were detected on chromosomes 2 (2H), 3 (3H), and 7 (5H), respectively. Rbgq1 and Rbgq2 have not been reported before, and did not map to a chromosome region where a major gene for powdery mildew had been reported. Four QTLs (Rrsq1, Rrsq2, Rrsq3 and Rrsq4) for resistance against scald were detected on chromosomes 3 (3H), 4 (4H) and 6 (6H). All four mapped to places where QTLs for scald resistance had been reported before in different populations.     

Race differentiation and search for sources of resistance to Rhynchosporium secalis in barley in Italy

Summary Barley in Italy has recently been seriously affected by Rhynchosporium secalis. The pathogenic variation of the fungus was studied and 17 races were differentiated on 13 barley cultivars carrying most of the currently known genes for resistance. RC 1, the most virulent and most frequent race, was virulent on 10 out of the 13 differentials and the remaining races proved to be less virulent variants of RC 1. Atlas (C.I. 4118), Atlas 46 (C.I. 7323) and Osiris (C.I. 1622) were the only three differentials resistant to all the analyzed single-spore isolates.Differential cultivars previously assumed to have identical resistance factors did not react in the same way to all the Italian races, thereby revealing either undisclosed differences in the genes described or the presence of additional unidentified ones.Our findings were compared with previous data about virulence of scald populations from different countries, on the basis of tests with common differentials: fundamental differences were found between the Italian population and those of other countries with regard to virulence patterns.The susceptible reactions to race RC 1 of most barley cultivars grown in Italy indicate the urgent need for resistance genes to be incorporated in the cultivated material. Seventy-one barley accessions, known as sources of resistance in different parts of the world, were screened for their behaviour to races RC 1 and RC 13. Twenty-two appeared resistant to both of them.     

Quantitative trait loci for scald resistance in barley localized by a non-interval mapping procedure

Three quantitative trait loci (QTL) for scald resistance in barley were identified and mapped in relation to molecular markers using a population of chromosome doubled‐haploid lines produced from the F₁ generation of a cross between the spring barley varieties ‘Alexis’ and ‘Regatta’. Two field experiments were conducted in Denmark and two in Norway to assess disease resistance. The percentage leaf area covered with scald (Rhynchosporium secalis) ranged from 0 to 40% in the 189 doubled‐haploid (DH) lines analysed. One quantitative trait locus was localized in the centromeric region of chromosome 3H, Qryn3, using the MAPQTL program. MAPQTL was unable to provide proper localization of the other two resistance genes and so a non‐interval QTL mapping method was used. One was found to be located distally to markers on chromosome 4H (Qryn4) and the other, Qryn6, was located distally to markers on chromosome 6H. The effects of differences between the Qryn3, Qryn4 and Qryn6 alleles in two barley genotypes for the QTL were estimated to be 8.8%, 7.3% and 7.0%, respectively, of leaf covered by scald. No interactions between the QTLs were found.     

Genes for resistance in barley to Finnish isolates of Rhynchosporium secalis

Summary Twenty Finnish isolates of Rhynchosporium secalis (Oud.) J.J. Davis, the causal agent of scald, were taken from infected barley (Hordeum vulgare L.) plants and inoculated on to seedlings of a differential series of barley containing a range of major genes for resistance to the fungus, as well as on to six Nordic 6-row spring barleys and three winter ryes (Secale cereale L.). These fungal isolates derived from four sites and three host varieties. Disease development was monitored on two leaves of seedlings in the greenhouse employing a standard scale, and on adult plants in the field by assessing the diseased area on the three uppermost leaves. A comparison was also made between the pathogenicity and virulence of ten Finnish and ten Canadian R. secalis isolates. The Finnish isolates varied in virulence, but with the exception of Algerian (CI 1179) seedlings and adult La Mesita (CI 7565) all seedlings and adult plants of the entire differential series were resistant to all isolates. Canadian isolates were, on average, less virulent than Finnish isolates. All the Nordic checks were susceptible to all Finnish and seven Canadian isolates, but differences in the degree of susceptibility were evident. Isolates of R. secalis from barley were non-pathogenic on rye, isolates from Elymus repens L. were non-pathogenic on barley and rye, and isolates from rye were only pathogenic on rye. Finnish R. secalis isolates contain no redundant pathogenic diversity. The differential series represents a useful, but as yet untapped, source of resistance to R. secalis for Finnish barley breeders.     

Characterization of resistance genes against scald (Rhynchosporium secalis (Oudem.) J.J. Davis) in barley (Hordeum vulgare L.) lines from central Norway, by means of genetic markers and pathotype tests

Rhynchosporium secalis is a serious pathogen of barley (Hordeum vulgare L.) in central Norway. A breeding effort was initiated in 1977 to introduce resistance from different sources into adapted genotypes, and the first cultivar from the program was recently released. However, little is known about the resistance genes introgressed in this cultivar or in advanced breeding lines. An effort was made to address this issue through a set of isolates and available molecular markers. Fourteen breeding lines and their resistance donors were investigated by evaluating their reactions to 11 R. secalis isolates. Bulked segregant analysis was used to identify molecular markers linked to resistance genes in 12 of the breeding lines. The isolates were found to be of less discriminating value than the markers. Useful information has been obtained as to the nature of several of the resistance genes introgressed. Eight of the 12 breeding lines contained introgressed genes that were located at the `complex Rh' locus on chromosome 3H and hence may not easily be pyramided into the same genotype. Previous information about the nature of the resistance in `Jet' is questioned. Neither of the resistance genes Rh or Rh2 seems to have been incorporated into Norwegian breeding material.     

Pathogenicity of the Rhynchosporium secalis population in the Western Cape province of South Africa

The virulence spectra of 50 Rhynchosporium secalis isolates from a population in the Western Cape province of South Africa were determined, and 21 races were detected when evaluated against 17 differential cultivars. The virulence spectrum of the R. secalis population shows considerable variation, and carries unnecessary virulence genes which is quite unexpected, since chiefly susceptible barley cultivars are grown in the south Western Cape. The two most prevalent races, namely races 4 and 7 had three and four virulence genes respectively. Both race 4 and 7 were virulent on the most susceptible cultivars, West China, Steudelli, C.I.8618 and C.I.2226. Considering the resistance genes reported for the cultivars Atlas 46, Turk, and C.I.3515 which showed no susceptible cultivar-pathogen interaction, it would appear that the Rh-Rh3-Rh4 complex is primarily involved in conferring resistance to the local R. secalis isolates This revised version was published online in July 2006 with corrections to the Cover Date.     

Identification of Resistance Genes Against Powdery Mildew in Four Accessions of Hordeum Vulgare SSP. Spontaneum

Summary Four newly detected accessions of wild barley (Hordeum vulgare ssp. spontaneum) resistant to powdery mildew caused by Blumeria graminis f. sp. hordei were studied with the aim of finding the number of genes/loci conferring the resistance of individual accessions, the type of inheritance of the genes and their relationships to the Mla locus. F₂ populations after crosses between the winter variety ‘Tiffany’ and four wild barley accessions and use of microsatellite DNA markers were focused on the identification of individual resistance genes/loci by means of their chromosomal locations. In PI466495, one locus conferring powdery mildew resistance was identified in highly significant linkage with the marker Bmac0213. This location is consistent with the known locus Mla on chromosome 1HS. In the other three accessions the resistance was determined by two independent loci. In PI466197, PI466297 and PI466461, one locus was identified on chromosome 1HS and three new loci were revealed on chromosomes 2HS (highly significant linkage with Bmac0134), 7HS (highly significant linkage with Bmag0021) and 7HL (significant linkage with EBmac0755). Our prospective aim is identification of further linked DNA markers and the exact location of the resistance genes on the barley chromosomes.     

Pathogenic variation among isolates of Rhynchosporium secalis from barley grass growing in South Eastern Australia

Summary The pathogenicity of 182 single spore isolates of Rhynchosporium secalis from Hordeum leporinum and 94 isolates from H. vulgare collected from throughout southeastern Australia was tested on 15 barley varieties, each having different combinations of resistance genes. Forty five percent of the barley grass isolates were pathogenic on 5 or more varieties but only 6% of the cultivated barley isolates attacked this range of varieties. On the basis of reaction type 20 different pathogenicity groups were recognised, with barley grass isolates being classified into 19 and the cultivated barley isolates into 5, four of which were the same as the barley grass isolates. Numerical analysis of data on leaf area damage inferred 33 groups, 24 of which were unique to barley grass isolates, two to cultivated barley isolates and 9 common to both groups. There was as much variation in pathogenicity among single spore isolates from the same lesion as between isolates from different lesions collected from the same or different locations.     

Evaluating resistance of barley breeding lines to scald in field plots

Summary A 0–4 scoring system to quantify scald (Rhynchosporium secalis) infection is suggested. Scores 1, 2, 3 and 4 allocated to represent 1/4, 1/2, 3/4 and 4/4 of the crop canopy scalded are easy to comprehend and intermediate scores e.g. 0.5, 1.5, 2.5 and 3.5 give it the breadth of a quantitative scale. Scores on a large number of lines showed a high degree of repeatability and were found to be highly correlated with the log transformed values of the actual leaf area damage. Although it was suggested that predictions of leaf area damage at scores 3–4 should be applied with caution, broad generalization of the scores in discriminating the amount of disease were shown to be soundly based and offered plant breeders a tool to standardize the evaluation of scald resistance in field plots on a large scale with this quick and reliable scoring system.     

Inheritance of resistance to Rhynchosporium secalis in spring barley (Hordeum vulgare L.)

The inheritance of durable resistance of selected spring barley varieties to Rhynchosporium secalis was investigated. Data from the F₂ generation of a 4 × 4 diallel, without reciprocals and the F₄ generation of three crosses selected out of this diallel, suggest that resistance in this sample of varieties tested is complex in inheritance. Significant additive effects were detected indicating that the resistance level of barley cultivars may be improved by the hybridisation of suitable varieties. However, the genes conferring resistance seem to be concealed by the expression of one completely dominant resistance gene in our set of varieties. These results are partly in conflict with previous results on the inheritance of resistance to R. secalis in the breeder's line ‘11258/2286₁₃A’ indicating that the effectiveness of this resistance gene may be greatly influenced by the genetic background of the current population of R. secalis.