Yield of doubled haploid lines of Nordic spring barley infected with net blotch, Pyrenophora teres

Five Nordic spring barley lines (‘Rolfi’, ‘Arve’, ‘Botnia’, ‘Pohto’ and WW7977) and doubled haploid (DH) populations from a half diallel of crosses between them, were sown in the field in Finland over 2 years and were artificially infected with Pyrenophora teres, the causal agent of net blotch. The purpose of the experiments was to determine the extent of yield loss under net blotch infection in a range of parent barleys and DH populations differing in symptom expression. Analysis of foliar damage symptoms, yield and aerial biomass data indicated that, in both years, there were statistically significant differences among parents and crosses, but the relationships between symptom expression and yield maintenance and between symptom expression and aerial biomass maintenance were stronger in 1997, when yields were higher and net blotch was less severe.     

Diallel analysis of net blotch resistance in doubled haploid lines of Nordic spring barleys

A half-diallel was made between five six-rowed Nordic spring barleys to study the genetics of resistance to net blotch. Twenty-five doubled-haploid (DH) lines from each cross and the parents were sown in hill plots in Finland in 1997 and 1998. The plots were artificially inoculated with Pyrenophora teres Drechs. f. teres Smedeg. and assessed for resistance to net blotch. There were statistically significant differences in resistance of the five parents to net blotch. General combining ability (GCA) of the parents and specific combining ability (SCA) effects in the progeny were statistically significant in both years, but GCA effects predominated. Evidence for additive epistasis was minimal. Progeny of a particular cross were less resistant to net blotch than the better parent. The most resistant progeny were derived from the cross between the two most resistant parents, Pohto and WW7977, and resistance was governed by at least eleven effective factors. Narrow sense heritability estimates for resistance to net blotch were high during both years (0.84–0.99). It appears that net blotch resistance of progeny from crosses can be largely predicted from reactions of the parents. Quantitative resistance to net blotch can be further advanced by identification and incorporation of superior parents, from a screening such as reported here, into a recurrent selection breeding programme. This revised version was published online in August 2006 with corrections to the Cover Date.     

Mapping seedling resistance to net form of net blotch (Pyrenophora teres f. teres) in barley using detached leaf assay

To develop molecular markers against Pyrenophora teres f. teres in barley, a detached leaf assay was conducted on two DH populations with a set of 11 single conidial lines (SCLs). Out of these, three showed different reactions in the DH population ‘Uschi × HHOR3073’ and two in ‘(P x V) × HHOR9484’. For SCL ‘QLB’, a 1r : 1s (χ² = 2.78) segregation was observed in the population ‘Uschi × HHOR3073’. In contrast to this, a continuous variation was observed for the SCL ‘WvB’ and ‘d8_4’ in the DH population ‘Uschi × HHOR3073’ and for ‘AR’ and ‘net1840’ in ‘(P × V) × HHOR9484’. With respect to resistance to the SCL ‘QLB’, a single major gene was located on chromosome 7H, and for resistance against SCL ‘WvB’, two QTLs were detected on chromosome 3H and 7H, and against SCL ‘d8_4’, two loci were mapped on chromosome 3HS in ‘Uschi × HHOR3073’. In the DH population ‘(P x V) × HHOR9484’, one locus conferring resistance to the SCL ‘AR’ was located on chromosome 3H. For resistance to SCL ‘net1840’, two QTLs were mapped on chromosome 4H and 5H.     

Radiation interception and radiation use efficiency of near-isogenic wheat lines with different height

Resistance to Pyrenophora teres Drechs. f. teres Smedeg., the net blotch pathogen, was studied in six 6-row Nordic spring barleys (Hordeum vulgare L.) in the field and in the greenhouse. The barley genotypes were: Arve, Agneta, Artturi, H6221, Pohto and WW7977. Disease progress was monitored in the field (1994 and 1995) in small artificially infected plots, sown at commercial seeding rates, and in infected hill plots (1994). Areas under the disease progress curves (AUDPC) and apparent infection rates (r) were calculated for the uppermost 3 or 4 leaves. Terminal severities (TS) were also recorded. Infection response of seedlings to a range of P. teres isolates was assessed in the greenhouse using a standard scale. In small plots in the field, Arve and Agneta were very susceptible to P. teres infection, as indicated by large values for AUDPC and TS. H6221 and WW7977 were highly resistant, while Artturi and Pohto were moderately resistant. In hill plots the situation was similar, except that Artturi and Pohto appeared less resistant than in the small plots. The relatively greater resistance of H6221 and WW7977 was reflected in seedling infection responses. According to the results of these experiments, H6221 and WW7977 possess adequate levels of quantitative resistance to P. teres to make them useful parents in future crossing programs aimed at improving net blotch resistance in Finnish spring barleys. This revised version was published online in July 2006 with corrections to the Cover Date.     

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.     

Inheritance of resistance to Pyrenophora teres f. teres in spring barley

The inheritance of seedling resistance to a Swedish isolate of Pyrenophora teres f. teres was investigated in four resistance sources of spring barley. Accessions CI 2330, CI 5791, CI 5822 and CI 9779 were used as resistance sources, and the cultivar ‘Alexis’ was used as a susceptible parent in different crosses. From the disease reaction in the F₁, F₂ and F₃ generations it was concluded that the resistance was governed by the same two complementary genes in CI 5791, CI 822 and CI 9776. One of these genes was present in CI 2330. The first three cultivars were highly resistant to the isolate used in this investigation. These results, when combined with earlier studies, suggest that CI 5791, CI 5822 and CI 9776 may be of great value as sources of resistance to barley net blotch. Spearman's rank correlation between the disease reaction of F₂ plants and their F₃ progeny was highly significant (r = 0.75; P ≥ 0.001) It is suggested that selection in the F₂ generation is effective. In a backcross breeding scheme, single plant reactions in F₁ or F₂ need to be confirmed in later generations.     

Reaction of European spring barley varieties to a population of the net blotch fungus

About 320 European spring barley varieties and about 40 spring barley lines were tested in the field for reaction to a local population of the spot form of the net blotch fungus Pyrenophora teres forma maculata with the imperfect stage Drechslera teres. The European barley varieties showed a wide range of disease reactions, from nearly resistant to very susceptible. At least three groups of genetically related barley varieties could be distinguished: (0)‘Agneta’ and ‘Clermont’, (2)‘Nordal’ and ‘Arve’, and (3)‘Tellus’,‘Pamina’, ‘Albert’ and ‘Birka’. The parentages of these three groups of barley are mutually distantly related and may thus constitute three different sources of resistance to the pathogen population used. The range of disease reaction is similar in varieties from different European countries but it differs among different breeding stations. The relatively high level of resistance is fairly uniformly distributed in contemporary barley varieties all over most of Western Europe. Over time, from about 1830‐1982, the average level of resistance varied around 4.0 (scale value) (from 2.8 to 4.8). However, since about 1940 the variation in disease reaction has become much wider (from 108 to 6.9) for unknown reason(s).     

Generation Mean Analysis of Inheritance of Resistance to Pyrenophora teres in Barley

F1, F2, Fl × parentl (BC1), and Fl × parent2 (BC2) generations resulting from four crosses among seven cultivars of barley used in national and international breeding programmes were tested at the seedling stage for their resistance to a mixture of five isolates of Pyrenophora teres. Four methods were used to assess disease resistance: infection type (IT), average lesion size (ALS), number of lesions per unit leaf area (NL) and percent leaf area infected (PLAI). Gene actions were estimated by generation mean analysis on each of the four crosses and on each of the evaluation methods. Significant additive and additive × additive epistatic effects were found. Infection type and percent leaf area infected were found to be highly correlated in all four crosses. These results suggest that barley breeders could improve the level of resistance to P. teres by making appropriate crosses between highly susceptible barley cultivars.     

QTL mapping of net blotch resistance genes in a doubled-haploid population of six-rowed barley

Net blotch, caused by Pyrenophora teres f. teres, is a damaging foliar disease of barley worldwide. It is important to identify resistance germplasm and study their genetics. 'Chevron', a six-rowed barley used as a parent for the production of a doubled haploid (DH) population for mapping of Fusarium head blight (FHB) resistance, was also found to be resistant to net blotch. To map the resistance genes, the population was evaluated for resistance at the seedling stage in a greenhouse. The resistance data showed a two-peak distribution. Through linkage mapping, one resistance gene, tentatively called Rpt, was located on chromosome 6HS flanked by Xksua3b-Xwg719d, which was also detected by QTL mapping. This QTL explained 64% of the phenotypic variance for the resistance in this DH population. In addition, a minor QTL was found on chromosome 2HS defined by Xcdo786-Xabc156a. 'Chevron' and 'Stander' contributed the resistant alleles of Rpt and the 2HS QTL, respectively. Both QTLs together explained nearly 70% of the phenotypic variance. The markers for these QTLs are useful for marker-assisted selection of net blotch resistance in barley breeding.     

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.     

Variation in Ethiopian barley landrace populations for resistance to barley leaf scald and netblotch

One-hundred and eighty landrace populations and six-hundred single-head plants selected from 60 promising populations were evaluated for resistance to scald and netblotch at three locations in Ethiopia. Each accession was tested with and without the application of 50% of the recommended rate of fertilizer at planting. Plants were rated for disease attack two to four times during the season. Both diseases were enhanced by the application of fertilizer and were more severe at the testing sites of Holetta and Bekoji than at Sheno. The difference in disease resistance among and within populations was considerable. Moreover, populations from Arsi and Bale tend to be more susceptible to scald but more resistant to netblotch than populations from other regions. Populations collected from higher altitudes were more resistant to scald, but susceptible to netblotch, than were populations from lower altitudes. The paper illustrates approaches to the identification of valuable genotypes from landrace populations that can be incorporated into a breeding programme for the development of improved varieties with resistance to the principal diseases of barley in Ethiopia.     

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.     

Genotype × environment interactions in the expression of net blotch resistance in spring and winter barley varieties

The net blotch resistance of barley varieties widely grown in Denmark was studied using data obtained from naturally infected field plots and inoculated disease nurseries in multiple years and locations. Data were analysed by combining two statistical procedures, 1) joint regression (JR) and 2) additive main effect, multiplicative interactive effect analysis (AMMI). Of the total variation for net blotch disease severity, 61–81% could be explained by plant genotype (G) and environment (E) main effects. Of the remaining variation, 77–86% could be explained by significant G × E interactions that were due to genotype sensitivity to the mean environmental disease level and/or to specific reactions of individual genotypes in particular environments. G × E interactions led to a different ranking of varietal performance across environments. There was evidence for G × E interactions caused by the virulence characteristics of the initial inoculum sources. The spring barley varieties Alexis and Bartok and the winter barley varieties Jolante, Ludo, and Rafiki were identified as varieties with high levels of net blotch resistance in all environments. Their performance was little affected by G × E interactions, suggesting that they represent good sources for non-specific resistance. Combining JR and AMMI reduced the dimensionality of complex G × E problems greatly, identified systematic reaction patterns of varieties possibly pertaining to different resistance mechanisms, and described specific and non-specific resistance of varieties by means of few parameters while maintaining the possibility to reconstruct the original data with little loss of information. This is very useful for improving the evaluation of varietal resistance for breeding and disease management purposes. This revised version was published online in July 2006 with corrections to the Cover Date.     

Approaches for field assessment of resistance to leaf pathogens in spring barley varieties

The resistance of spring barley varieties to powdery mildew, leaf rust, leaf scald and net blotch was characterized by using results from inoculated small‐plot nurseries and larger survey plots subject to natural infection. The experiments were conducted in different environments. Both trial types often yielded complementary results with respect to the ranking of varieties suggesting that a recommended variety characterization should include both naturally infected survey‐type trials and nursery trials in which the most relevant pathogen isolates and/or isolate mixtures or populations are used for inoculation. Average and median values of the diseased leaf area of a variety were highly correlated with each other and with the ‘genotype main effect’ determined by joint regression analysis, whereas maximum diseased leaf area was poorly correlated with them. Statistics based on absolute disease severity values were highly correlated with the corresponding statistics derived from relative values. It is suggested that one should use at least two parameters to characterize the disease resistance of a variety, a parameter indicating the overall resistance level and a parameter indicating the potential susceptibility and/or resistance instability of a variety. For practical purposes, the genotype median and maximum, respectively, may represent these, although statistically more appropriate parameters do exist.     

Haplotype structure around the nud locus in barley and its association with resistance to leaf stripe (Pyrenophora graminea)

The naked/hulled kernel trait is controlled in barley by a single gene called nud, on chromosome 7H. The first aim of this work was use bulked segregant analysis to find, new PCR‐based markers linked to nud for marker‐assisted selection (MAS). A new SCAR marker (sJ14) was developed, which is useful for introgressing the naked trait. This, and three other SCARs, were placed on the ‘Proctor’ × ‘Nudinka’ map to detail a 0.9‐cM fragment tagging nud. In order to evaluate the haplotypes around the nud locus, a phenotypically differentiated collection of naked/hulled genotypes was characterized by means of the above markers. Eight different marker haplotypes were found in the breeding germplasm, and a new allele for the marker sKT7 was found. The same barley collection has been surveyed for resistance/susceptibility to leaf stripe (Pyrenophora graminea), in order to investigate any possible association between this and other traits. The naked/hulled seed trait was not associated with resistance/susceptibility to the fungus.     

大麦种质资源成株期抗旱性鉴定及抗旱指标筛选

干旱是影响大麦生产的主要因素之一。在鉴定大麦种质资源成株期抗旱性的基础上,筛选抗旱指标,可为培育抗旱品种提供依据。本研究在2016和2017年在大麦生长主要需水期平均降雨量不足40 mm的甘肃省武威市进行田间试验,以30份大麦种质资源为研究对象,设置正常灌水和干旱胁迫处理,测定大麦株高、穗长、分蘖数、单株粒数、单株粒重、穗粒数、千粒重和产量,采用抗旱性度量值(D)、综合抗旱系数(CDC)、加权抗旱系数(WDC)、相关分析、频次分析、主成分分析、灰色关联度分析、隶属函数分析、聚类分析和逐步回归分析相结合的办法,对其进行抗旱性鉴定及抗旱指标的筛选。干旱胁迫对测定的各指标均有极显著影响。频次分析表明,各指标对干旱胁迫反应的敏感程度依次为产量、株高、单株粒重、穗长、单株粒数、分蘖数、穗粒数和千粒重。相关分析表明,产量与株高、穗长、分蘖数、单株粒数和单株粒重呈极显著正相关,与穗粒数呈显著正相关、与千粒重呈不显著正相关。主成分分析表明,5个主成分可以代表大麦抗旱性86.39%的原始数据信息量。基于D值、CDC值和WDC值的大麦种质资源抗旱性排序基本一致。灰色关联度分析表明,各指标单项抗旱系数值与D值间的关联度依次为产量、单株粒重、单株粒数、穗长、株高、分蘖、穗粒数和千粒重,各指标DC值与WDC值间的关联大小依次为单株粒重、产量、单株粒数、分蘖、穗长、穗粒数、株高和千粒重。根据D值进行聚类分析,将供试大麦种质资源依据抗旱性从强到弱分为5个抗旱等级,其中1级5份、2级1份、3级11份、4级10份、5级3份。除分蘖和千粒重外,其余指标的隶属函数值、CDC值、D值和WDC值均随着抗旱级别的升高而增大。回归分析表明, 5个测定指标均与D值密切相关。干旱胁迫对大麦种质资源成株期各指标均有极显著影响。确定了D值为适宜的抗旱鉴定指标。筛选出成株期抗旱性强的大麦种质资源甘啤7号、Z06-278-9、MERIT、NEVADA和西藏25,可为大麦抗旱育种、抗旱机制以及干旱调控缓解机制的研究提供技术支持。穗长、单株粒数、单株粒重、穗粒数、产量可作为大麦种质资源成株期简单、直观的抗旱评价指标。     

Isolation, identification and characterization of disomic and translocated barley chromosome addition lines of common wheat

Two disomic barley chromosome addition lines and five translocated chromosome addition lines of common wheat cultivar Shinchunaga were isolated. They were derived from a hybrid plant between Shinchunaga and cultivated barley Nyugoruden (New Golden) by backcrossing with wheat and self pollination. Barley chromosomes added to chromosome arms involved in the translocated chromosomes were identified by C-banding method and by crossing these lines with Chinese Spring/Betzes addition lines. Two disomic addition lines were identified to have chromosome 6 and 7 of barley, respectively. Two of the five translocated chromosome addition lines were clarified to have same chromosome constitution, 42 wheat chromosomes and a pair of translocated chromosomes constituted with a long arm of chromosome 5B of wheat and a short arm of chromosome 7 of barley. The other three lines could not be identified due to chromosome rearrangement. Performances of these seven lines on agronomic characters were examined. Addition of barley chromosome 7 induced early heading, and chromosome 6 showed lated heading. Almost all of the lines except that of chromosome 6 showed short culm length and all showed reduced number of tillers, spikelets and grains per ear, and low seed fertility. These lines would be useful for genetic analyses in wheat and barley and for induction of useful genes of barley into wheat. This revised version was published online in July 2006 with corrections to the Cover Date.     

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.     

EcoTILLING for the identification of allelic variation in the powdery mildew resistance genes mlo and Mla of barley

In this investigation the successful implementation of a CEL I‐based mutation detection technique for the discovery and detection of DNA polymorphism in the genes mlo and Mla of barley is described. The technique is called EcoTILLING, which is a high‐throughput method to detect and discover new point mutations and small insertions/deletions in DNA. That the method not only reveals polymorphism between different alleles but can also be used as a powerful genetic marker is demonstrated. The genes mlo and Mla are involved in the defence of barley against the fungal pathogen powdery mildew. The powdery mildew resistance gene mlo is a single copy gene, whereas multiple alleles exist at the Mla locus. With EcoTILLING it was possible to identify point mutations and deletions in each of the 11 mlo mutants tested. For Mla 25 natural barley variants were tested, and although the identification was complex due to the presence of highly similar paralogues of Mla, most of the recently identified alleles from Hordeum vulgare ssp. spontaneum were identified. This method offers the possibility to combine different mlo alleles with different Mla alleles from wild barley to obtain cultivars with more durable resistance.