Stable resistance in barley to Pyrenophora teres f. teres isolates from the Nordic-Baltic region after increase on standard host genotypes

Results from tests of a mixture of Finnish net blotch, Pyrenophora teres Drechs. f. teres Smedeg., isolates on a differential series of barley seedlings, comprising 17genotypes, indicated that patterns of infection response (IR)and percentage leaf area damaged (PLAD) were unaffected by differences in seedling size. Variation of the concentration of inoculum between 1,250 conidia ml^-1 and 20,000 conidia ml^-1 produced similar patterns of IR and PLAD on the differential series. IR and PLAD scored on the second seedling leaf differentiated resistance to P. teres f. teresamong the genotypes better than on the first seedling leaf. Ina second experiment, 120 single-spore P. teres f. teres isolates from Finland, Sweden, Norway, Latvia, Estonia and Ireland were used in tests conducted in the greenhouse to differentiate them in terms of virulence reaction on seedlings of six differential barley genotypes. Each isolate was tested directly following isolation from the leaf material and after having passaged each through barley cvs. Arve or Pohto, to produce 360 isolates in total. Virulence of the isolates differed significantly on the members of the differential series, but differences associated with country of origin and passaging, and interactions, were small. It is concluded that little variation between virulence of P. teres f. teres isolates is evident over a large geographic area, incorporating Nordic and Baltic countries, and Ireland. Barley genotype response to P. teres f. teres appeared to be of more significance than relative virulence of the pathogen isolates. This could simplify breeding barley for improved resistance to this phytopathogen. This revised version was published online in August 2006 with corrections to the Cover Date.     

Resistance of barley genotypes to brown leaf spot

Resistance of 59 barley cultivars to three distinct isolates of Bipolaris sorokiniana, causing brown leaf spot was tested on seedlings and adult plants in a screen house and in a growth room, by determination of disease incidence, disease severity, and rate of lesion expansion. The disease reaction and severity at the seedling stage differed among cultivars. The cultivars MNS Brite, NDB 112, Kindred, Parkland, and Oderbrucke were moderately resistant to brown leaf spot, but the resistance was partial to all three isolates. None genotype with complete resistance was found. The cultivars CI 9539, Svanhals, and Chevron, considered as sources of resistance, were classified as moderately susceptible, and susceptible, respectively. Disease severity, disease reaction, and incidence of B. sorokiniana in grains also differed among the cultivars at the adult plant stage. Nevertheless, none of the genotypes was resistant to B. sorokiniana. The rate of lesion expansion differed among cultivars BR 2, MN 698, Kitchin, Svanhals, and NDB 112, with NDB 112 having the slowest expansion rate (0.194–0.205 mm²/day).     

Diversity among Finnish net blotch isolates and resistance in barley

Summary Seedlings of a differential barley (Hordeum vulgare L.) series (21 genotypes) and of six check genotypes were used in the greenhouse to assess variation in virulence among 20 single-spore isolates of the net blotch pathogen. Pyrenophora teres Drechs. f.teres Smedeg., collected from various sites in Finland. The experiment was run twice and symptom expression was recorded on the first three leaves. Analysis of second leaf symptom scores from Run 1 indicated differences in virulence between isolates, all of which were pathogenic, and differential resistance among the barleys. The virulence of P. teres isolates appeared to be conditioned by the host barley from which the isolate derived; the average virulence of isolates collected from a susceptible host was greater than that of isolates collected from a less susceptible host. Results from Run 2 were similar regarding resistance within the barleys, but variation in virulence among the P. teres isolates was not consistent with that from Run 1. CI 9819 caries duplicate genes for resistance to P. teres (Rpt1b and Rpt1c), and CI 7548 possesses Rpt3d. Both genotypes were highly resistant to all isolates. The Rpt1a gene of Tifang (CI4407) conferred resistance to all isolates in Run 2, but only to about half of the isolates in Run 1. The checks, including two of the symptomatically most resistant Nordic barley genotypes, were universally susceptible during these stringent tests. No selective pressure has been placed on Finnish isolates of P. teres through previous deployment of major resistance genes, and it is speculated that any variation in virulence among isolates is likely to be due to a combination of evolutionary forces including, natural selection, random genetic drift and gene flow.     

Localization of a resistance gene and identification of sources of resistance to barley leaf stripe

In order to determine more precisely the location of the barley leaf stripe gene, called the ‘Vada-resistance gene’, on barley chromosome 2, 63 chromosome-doubled barley lines were tested. Using data on known chromosome 2 genetic markers, the ‘Vada-resistance gene’ was estimated to be located between the markers MSU21 and Xris45b, and at a distance of about 20% recombination from the powdery mildew resistance gene MILa. We suggest that the ‘Vada-resistance gene’ is designated Rdg1a and that all former leaf stripe resistance gene designations should be rejected. To identify possible new sources of resistance, 11 barley cultivars/lines known to possess leaf stripe resistance and originating from different parts of the world, were tested with one Danish and two Syrian isolates of the leaf stripe fungus. Three apparently genetically different sources of race-specific resistance were found. The ‘Vada-resistance’ in the cultivar ‘Golf was effective against seven out of eight isolates’ populations of the leaf stripe fungus differing in geographical origin.     

Postulation of leaf-rust resistance genes in Czech and Slovak barley cultivars and breeding lines

Leaf‐rust resistance (Rph) genes in 61 Czech and Slovak barley cultivars and 32 breeding lines from registration trials of the Czech Republic were postulated based on their reaction to 12 isolates of Puccinia hordei with different combinations of virulence genes. Five known Rph genes (Rph2, Rph3, Rph4, Rph7, and Rph12) and one unknown Rph gene were postulated to be present in this germplasm. To corroborate this result, the pedigree of the barley accessions was analysed. Gene Rph2, as well as Rph4, originated from old European cultivars. The donor of Rph3, which has been mainly used by Czech and Slovak breeders, is ‘Ribari’ (‘Baladi 16’). Rph12 originates from barley cultivars developed in the former East Germany. Rph7 in the registered cultivar ‘Heris’ originates from ‘Forrajera’. A combination of two genes was found in 10 cultivars. Nine heterogeneous cultivars were identified; they were composed of one component with an identified Rph gene and a second component without any resistance gene. No gene for leaf rust resistance was found in 17 of the accessions tested. This study demonstrates the utility of using selected pathotypes of P. hordei for postulating Rph genes in barley.     

Genome-wide association studies in a barley (Hordeum vulgare) diversity set reveal a limited number of loci for resistance to spot blotch (Bipolaris sorokiniana)

Spot blotch caused by Bipolaris sorokiniana is an important disease in barley worldwide, causing considerable yield losses and reduced grain quality. In order to identify QTL conferring resistance to spot blotch, a highly diverse worldwide barley set comprising 449 accessions was phenotyped for seedling resistance with three isolates (No 31, SH 15 and SB 61) and for adult plant resistance at two locations (Russia and Australia) in two years. Genotyping with the 50 k iSelect barley SNP genotyping chip yielded 33,818 informative markers. Genome-wide association studies (GWAS) using a compressed mixed linear model, including population structure and kinship, revealed 38 significant marker-trait associations (MTA) for spot blotch resistance. The MTA corresponded to two major QTL on chromosomes 1H and 7H and a putative new minor QTL on chromosome 7H explaining between 2.79% and 13.67% of the phenotypic variance. A total of 10 and 14 high-confidence genes were identified in the respective major QTL regions, seven of which have a predicted involvement in pathogen recognition or defence.     

Virulence studies of Swedish net blotch isolates (Drechslera teres) and identification of resistant barley lines

Six Swedish and one Canadian single spore isolate of Drechslera teres f. teres were used to screen 109 barley lines for disease resistance and to select a differential set of barley lines for use in assessing pathogen virulence. A large variation for net blotch resistance was found among the 109 barley lines which were classified into four groups, those showing: 1) only resistant reactions; 2) differential reactions; 3) only intermediate reactions and 4) only susceptible reactions. The European commercial varieties included, showed susceptibility to all Swedish isolates, but a few were resistant to the Canadian isolate. The 18- member differential set separated 25 Swedish and two Canadian isolates of D. teres into 14 pathotypes, three of which made up 59% of the isolates. Only one barley differential (CI 9776) was resistant to all net form isolates. Host selection on the pathogen seems to be present as all six isolates obtained from cv. Golf belonged to the same pathotype and 4 of 5 isolates from cv. Karin shared the same virulence pattern. The net form of net blotch (D. teres f. teres) predominated in the sampled regions and only one of 26 Swedish isolates was of the spot form (D. teres f. maculata). This revised version was published online in July 2006 with corrections to the Cover Date.     

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.     

Rapid phenotyping of adult plant resistance in barley (Hordeum vulgare) to leaf rust under controlled conditions

Breeding for adult plant resistance (APR) is currently impeded by the low frequency of annual field‐based testing and variable environmental conditions. We developed and implemented a greenhouse‐based methodology for the rapid phenotyping of APR to leaf rust in barley to improve the efficacy of gene discovery and cloning. We assessed the effects of temperature (18 and 23°C) and growth stage (1–5 weeks) on the expression of APR in the greenhouse using 28 barley genotypes with both known and uncharacterized APR. All lines were susceptible in week 1, while lines carrying Rph20 and several with uncharacterized resistance expressed resistance as early as week 2. In contrast, lines lacking Rph20 and carrying either Rph23 and/or Rph24 expressed resistance from week 4. Resistant phenotypes were clearest at 18°C. A subset of 16 of the 28 lines were assessed for leaf rust across multiple national and international field sites. The greenhouse screening data reported in this study were highly correlated to most of the field sites, indicating that they provide comparable data on APR phenotypes for screening purposes.     

Relationship between Frost Tolerance and Cold-Induced Resistance of Spring Barley, Meadow Fescue and Winter Oilseed Rape to Fungal Pathogens

Plants exposed to one stress factor may become more tolerant to another. Cold is the most often documented factor inducing plant resistance to pathogens. The aim of this work was to investigate whether resistance of spring barley and meadow fescue to Bipolaris sorokiniana and resistance of winter oilseed rape to Phoma lingam induced at 5 °C for 2, 4 or 6 weeks are associated with frost tolerance, water potential and soluble carbohydrate content. Cold‐acclimated plants of each species showed increased resistance to the studied pathogens. Barley, fescue and rape plants demonstrated higher frost tolerance after hardening, but only in the case of fescue a correlation between resistance to frost and resistance to B. sorokiniana was found. A significant decrease in the water potential of leaf cells was observed in cold‐acclimated barley and fescue. In these two species, water potential greatly affected resistance to B. sorokiniana. However, only in barley did accumulation of fructose, glucose and sucrose correlate as well with changes in water potential as with cold‐induced resistance to the pathogen. In the case of hardened rape, no correlation between the studied parameters was found. The results obtained indicated that the temperature of 5 °C used during cold acclimation was not favourable for hardening of this plant species.     

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.     

The Yd2 gene and enhanced resistance to barley yellow dwarf virus (BYDV) in winter barley

A breeding programme was developed to obtain barley yellow dwarf virus (BYDV)-resistant winter genotypes using the Yd2 gene. The aim was to incorporate the Yd2 allele into the new high-yielding genotypes to release cultivars that allow barley cultivation in areas where BYDV is endemic. The resistant lines were developed using pedigree selection. An ICARDA resistant line (83RCBB130) carrying the Yd2 gene was crossed with three susceptible, high-yielding winter varieties and their F₁ lines were either selfed or backcrossed to the matching susceptible parent. The best lines selected from subsequent selfing generations were evaluated in replicated trials in the presence or absence of BYDV, starting from F₆ and BC₁F₅ to F₈ and BC₁F₇ generations. Four genotypes with superior agronomic traits and BYDV resistance were selected.     

Utilization of exotic germplasm in Nordic barley breeding and its consequences for adaptation

Summary Utilization of exotic germplasm is one way to broaden genetic variation in breeding populations. This approach has recently been adopted in Sweden and Finland, where experimental barley populations has been established for research and pre-breeding purposes. The aim of the project is threefold: (1) to increase overall genetic diversity of Nordic barley breeding material; (2) to develop breeding material which possesses a high level of resistance for various barley diseases; and (3) to study effects of exotic germplasm on adaptation and agronomic performance. Both the Finnish and the Swedish barley populations include the same exotic material i.e. unadapted landraces from different parts of Asia and wild barley (Hordeum vulgare ssp. spontaneum) accessions. Locally adapted high-yielding barley lines were included in the populations. The establishment of these populations involved six crossing generations in order to promote recombination and enhance the break-up of linkage blocks. The paper discusses the third aim of the project. Studies on agronomic performance and adaptation showed that (1) agronomically valuable genotypes can be constructed through recombination using exotic germplasm for Nordic conditions, (2) that incorporation of exotic material is most successful when made in a local genetic base and (3) that exotic germplasm has an effect on adaptation.     

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.     

Sources of Resistance to Powdery Mildew in Barley Lines Derived from Hordeum spontaneum Collected in Israel

Fourty two barley lines direved from the F₇ of crosses between barley cultivars and different accessions of Hordeum spontaneum collected in Israel and 30 lines or varieties with known genes for resistance to powdery mildew were included m this study. Eleven European and three Israeli powdery mildew cultures, possessing virulence genes corresponding to known resistance genes, were used to make comparisons between the varieties with known resistance genes and H. spontaneum derived lines. The reaction pattern of 39 H. spontaneum derived lines was clearly different from the reaction pattern o; any of the known genes for mildew resistance included in this study. Only two cases were observed in which the reaction pattern of H. spontaneum derived lines agreed with reaction patterns of known genes for mildew resistance viz. Ml-a9 and Ml-p. Trie Mildew resistance of one line apparently traces back to uncontrolled outcrossing with a Ml-a.6+Ml-g resistant cultivar. Since the majority of the 42 host genotypes tested showed distinctive variation in resistant reaction types against different mildew cultures, this study docs not support the assumption that differences in resistant infection types against distinct mildew cultures are sufficient to indicate the presence of supplementary genes for resistance in a given genotype of the host. The results justify the conclusion that the natural population of H. spontaneum in Israel forms a large gene pool for mildew resistance which is not yet used m cultivated barley.     

Resistance to powdery mildew in selections from Moroccan barley landraces

Seventy-five barley landraces from Morocco were tested for resistance to powdery mildew and a number of different resistance genes were detected. Thirty-five isolates of Blumeria graminis f. sp. hordei and the Pallas isoline differential set were used. Isolates used in the experiment had virulences corresponding to all major resistance genes used in Europe. Forty-four of the tested landraces showed resistant reactions. From each of these landraces, one to five resistant plants were selected and 92 single plant lines were created. Six lines selected from 3landraces were assumed to carry the mlo gene but they were discarded after microscopic investigation. Seventeen lines were tested in the seedling stage with 17isolates and another 69 lines were tested with 23 differential isolates. These lines showed 71 reaction spectra to isolates of powdery mildew. Eight lines (9%), 255-3-3, 282-3-4, 286-1-1, 294-2-3,294-2-4, 295-1-2, 308-1-2 and 327-2-1, selected from 7 landraces showed resistance to all isolates. Seventy-eight lines (90%) showed a resistant infection type 2with more than 50% of the isolates used. In most of the selected lines (86%) unknown genes, alone or in combination with known specific resistance genes, were detected. Four different resistance alleles (Mlat, Mla6, Mla14 and Mla1) were postulated to be present in the tested lines. The most common was Mlat, which was postulated in 35 (41%) lines. The use of newly identified sources of powdery mildew resistance in barley breeding is discussed. This revised version was published online in August 2006 with corrections to the Cover Date.     

Relationship of Carbon Isotope Discrimination to Water Use Efficiency and Productivity of Barley Under Field and Greenhouse Conditions

This study was conducted to evaluate the application of carbon isotope discrimination (CID) as a selection criterion for improving water use efficiency (WUE) and productivity of barley (Hordeum vulgare L.) under field and drought‐stress conditions in a greenhouse. A total of 54 genotypes were screened for variability in CID under field conditions, while 23 genotypes were evaluated under water‐deficit conditions in the greenhouse. A survey of leaf CID of 54 genotypes at two field locations showed more than 2.14‰ difference between extreme genotypes. Significant (P ≤ 0.05) genotypic variation was found in WUE and CID that had a negative strong correlation. There was a negative correlation between leaf CID and aerial biomass in the greenhouse and among six‐row genotypes in the field. Correlations between leaf CID across field locations and across irrigation regimes in the greenhouse were significant (experiment 1, r = 0.79 and 0.94 for six‐ and two‐row genotypes), suggesting stability of the CID trait across different environments. Overall, these results indicate the potential of leaf CID as a reliable method for selecting for high WUE and productivity in barley breeding programmes in the Canadian prairies. Further work is currently underway to determine heritability/genetics of leaf CID and application of molecular marker‐assisted selection for the traits in barley breeding programmes.     

Adaptation of poplar leaf rust to Eastern cottonwood

Summary Genotypes of eastern cottonwood, Populus deltoides, from Kentucky or Mississippi were inoculated using a leaf disk assay with isolates of poplar leaf rust, Melampsora medusae f. sp. deltoidae (Mmd), from both locations to determine if pathogenic races differ at these locations. Disease severity measurements, including infection probability (number of uredia produced per uredospore), sporulation (number of uredospores/cm²), and the progeny/parent ratio (number of daughter uredia produced per mother uredium) were significantly higher when trees of southern origin were inoculated with isolates from southern locations as compared to isolates from northern locations. In addition, larger variation in disease severity measurements attributable to specific interactions, i.e. the variance due to isolates adjusted for virulence on a universal suscept, were found in northern than in southern isolates. These results suggest that isolates of southern origin have broader adaptation and are more virulent on local southern host genotypes than isolates of northern origin. However, the larger variance attributable to specific interactions in northern isolates indicates the potential for pathogen adaptation to resistant host genotypes in Kentucky.Journal series paper 92-8-81 of the University of Kentucky Agricultural Experiment Station     

Resistance spectra of wheat genotypes and virulence patterns of <Emphasis Type="Italic">Mycosphaerella graminicola</Emphasis> isolates in Iran

Co-evolution of wheat and its devastating pathogen Mycosphaerella graminicola (anamorph Septoria tritici), the causal agent of septoria tritici blotch, a foliar disease of wheat, is suggested to occur in Fertile Crescent as their center of origin and, thus, interaction between pathogen virulence and host resistance is important subject to be addressed. We have investigated resistance spectra of 54 wheat genotypes including a set of differentials carrying known resistance genes and virulence patterns of 14 M. graminicola isolates at seedling stage under controlled environmental conditions. The isolates were collected in Iran from five provinces. Diversity in virulence and aggressiveness was observed among the isolates from four provinces. Isolates collected from Golestan province were virulent to all wheat genotypes from germplasm of Iran, while specific resistances were identified to the isolates from other provinces. Among wheat genotypes, cvs. Chamran, Morvarid and Hirmand had the greatest number of specific resistances as well as partial resistance. Wheat genotypes of the differential set also differed in their reactions to the isolates. Arina, Flame and TE 9111 were specifically resistant to the greatest number of isolates from different provinces. Most isolates were virulent to the other differentials such as cvs. Shafir, Estanzuela federal and Courtot indicating that extensive adaption of virulence to most of the known resistance genes (Stb) has occurred in these regions. The new sources of resistance to highly virulent isolates from Iran may also be utilized in wheat breeding programs to develop resistant cultivars against pathogen populations in other countries.     

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.