Diallel analysis of partial resistance to powdery mildew caused by Erysiphe graminis f. sp. hordei in spring barley (Hordeum vulgare L.)

Summary Six spring barley genotypes with growth stage dependent expression of partial resistance to powdery mildew were crossed following a half diallel scheme. Data for percentage infected leaf area at different growth stages from the F1 and F2 generation and the F2 offspring were analyzed. The absence of a contrast between parents and offspring indicated that dominance effects were not important at the young growth stages. Diallel analysis showed that general combining ability effects (GCA) were important at all growth stages. Although significant, specific combining ability effects were of minor importance. The cross between the most resistant parents with the largest, negative GCA resulted in the most resistant progeny.     

Inheritance of resistance to powdery mildew (Erysiphe graminis f. sp. hordei) in eleven primitive barley varieties

The inheritance of the powdery mildew resistance of eleven primitive barley varieties was investigated. crosses with varieties with identified resistance genes revealed that at least three different genes are present in a group of six genotypes from Greece (Hor736, Hor847, Hor878, Hor1159, Hor1379 and Hor1873) and one from Turkey (Hor1188). One dominant gene is common to these seve genotypes and in the Greek accessions a second gene is present. The expression of this second gene was respectively dominant in the crosses with Hor736 and Hor1159, recessive in the crosses with Hor878 and Hor1873, and depended on the employed powdery mildew isolate in the crosses with Hor847 and Hor1379. The detected genes differ from the resistance genes of the crossing partners, viz. Ml-al2, Ml-(Ab), Ml-(CP) and Ml-(1402). The accessions from China (Hor824 and Hor4021) each possess two resistance genes which differ from Ml-a12, Ml-(La) and Ml-(CP). The major gene is common to both accessions and is either identical, allelic or closely linked to a gene in the variety Nigrate. The Columbian accession Hor1894 possesses one resistance gene linked to a resistance gene in Nigrate but this gene differs from the Ml-a locus. The variety Palestine (Hor3997) possesses two resistance genes of which one is allelic or closely linked to Ml-(at).     

Genetic analysis of powdery-mildew (Erysiphe graminis f.sp. hordei) resistance derived from wild barley (Hordeum vulgare ssp. spontaneum)

The inheritance of resistance to powdery mildew was investigated in 20 accessions of Hordeum spontaneum and in 20 F₄ lines derived from crosses between the variety ‘Aramir’ and 13 accessions of H. spontaneum. Two resistance genes were detected in 17 accessions, and three resistance genes in one accession. In two accessions, only one resistance gene was present. The 20 breeding lines showed a large variation in infection type and infection level. The genetic relationship between the resistance genes detected was investigated in the seven most resistant F₄ lines. These F₄ lines were divided into three groups which carried different resistance genes. In two lines, the detected resistance gene was shown to be race-specific.     

Effect of powdery mildew (Erysiphe graminis f.sp. hordei) on photosynthesis and grain yield of partially resistant genotypes of spring barley (Hordeum vulgare L.)

The effect of powdery mildew on the photosynthesis and grain yield of partially resistant spring barley was investigated. The effect of powdery mildew on the CO₂ exchange rate (CER) of inoculated and non-inoculated leaves was measured in several glasshouse experiments. Grain yield reduction by powdery mildew was determined in three field experiments. The CER of the inoculated leaves was reduced by infection but that of the non-inoculated leaves was not significantly different from that of the corresponding leaves of the control plants. The reduction in CER of the different genotypes was closely related to the percentage leaf area infected with powdery mildew. The correlation between degree of mildew infection and grain yield reduction varied between r = 0.84 (significant at P = 0.01) and r = 0.41 (not significant). Differences in greenness of the crop were correlated better with grain yield reduction (r = 0.66, P = 0.01) than powdery-mildew infection itself. Low percentages of powdery-mildew infection did not lead to significant reduction in grain yield. Partial resistance to powdery mildew could thus be a valuable attribute of future barley varieties.     

Detection of tolerance of barley cultivars to infection by powdery mildew (Erysiphe graminis f.sp. hordei)

Summary Barley genotypes representing a wide range of resistance expressions and origins, from major resistance genes in modern cultivars to field resistances in landraces, were assessed for tolerance to disease under glasshouse and field conditions. A few genotypes were picked out as showing less yield loss than would be expected from the level of mildew infection. Genotypes showing more than the expected yield loss were also found. The potential use of tolerance as a breeding character is discussed.     

Association of leaf rust and powdery mildew resistance in a recombinant derived from a Hordeum vulgare × Hordeum bulbosum hybrid

While studying powdery mildew resistance in a recombinant line (code 81882) derived from a Hordeum vulgare (cv. ‘Vada’) ×Hordeum bulbosum hybrid, a low infection type of resistance to leaf rust was observed. To determine the mode of inheritance of the leaf rust resistance and whether there was linkage between the two resistances, F₂ and F₃ progenies from crosses between 81882 and ‘Vada’ were inoculated with the leaf rust and powdery mildew pathogens. Southern blots were prepared using restricted DNA extracted from leaves of 82 F₂ plants and four chromosome 2HS sequences were hybridized with the blots to define the length of the introgression. The leaf rust resistance appears to be inherited as a single dominant gene on chromosome 2HS, which co-segregates with the powdery mildew resistance. There was an almost complete association between the resistances and the respective molecular markers, but it is likely that the strong linkage results from the frequent inheritance of the introgressed H. bulbosum DNA as an intact segment of chromatin with only low levels of recombination within the segment.     

小麦新种质CH1357抗白粉病遗传分析及染色体定位

白粉病是影响小麦产量和品质的一种主要病害。小偃麦衍生品系CH1357对白粉病具有较好的成株抗性,苗期对27个菌株表现为免疫或高抗,是一个高抗白粉病的优异抗源。为了明确其抗白粉病基因在染色体上的位置,对台长29/CH1357和绵阳11/CH1357的F_1、BC_1及F_(2:3)家系进行了遗传分析,并利用分离群体分组分析法(bulked segregantanalysis,BSA)将其初步定位。CH1357的白粉病抗性受1对显性核基因控制,位于染色体5DS,暂命名为PmCH1357。其侧翼连锁标记为Xcfd81和Xbwm8,在2个作图群体台长29/CH1357和绵阳11/CH1357中的遗传距离分别为2.0 cM/11.3 cM和1.5 cM/8.9 cM。PmCH1357与5DS染色体上已报道的其他抗白粉病基因抗谱不同,可能是一个新的抗源。     

Involvement of polyamines, diamine oxidase and polyamine oxidase in resistance of barley to Blumeria graminis f. sp. hordei

Polyamine levels and diamine and polyamine oxidase activities have been investigated in the first leaves of barley (Hordeum vulgare L.) in the absence of or following inoculation with conidia of powdery mildew (Blumeria graminis f. sp. hordei). Two cultivars with varying sensitivity to powdery mildew, viz., Chariot (resistant) and Golden Promise (susceptible) were used. The levels of putrescine, spermidine and spermine were found to be higher in the leaves of Chariot than in the leaves of Golden Promise and, with the exception of spermine, were generally higher in both cultivars after inoculation. In inoculated leaves of Chariot, levels of putrescine and spermidine peaked at 9 days and 12 days, respectively. In controls (uninoculated leaves), the activities of these enzymes, and putrescine and spermidine levels also increased but not to the same extent as in inoculated leaves. With Golden Promise, the levels of putrescine and spermidine in the inoculated leaves changed very little over sampling times but were always higher than in the controls. In this cultivar, there was little difference between inoculated leaves and the controls in diamine oxidase activity which reached a maximum value at 9 days post-inoculation. Activity of the bound form of diamine oxidase was low in both the cultivars. Polyamine oxidase was not detected at 3 days after inoculation in either cultivar but activity at fairly low levels was recorded at later times, usually reaching a maximum value at 9 days. The results suggest that polyamine metabolism and diamine oxidase activity in particular may be involved in the mechanism conferring resistance to barley powdery mildew in Chariot.     

Cloning, genetic and physical mapping of resistance gene analogs in barley (Hordeum vulgare L.)

The majority of verified plant disease resistance genes isolated to date belong to the NBS‐LRR class, encoding proteins with a predicted nucleotide binding site (NBS) and a leucine‐rich repeat (LRR) region. Using degenerate primers, designed from the conserved motifs of the NBS region in tobacco N and Arabidopsis RPS2 genes, we isolated 190 resistance gene analogs (RGA) clones from barley genomic DNA. A total of 13 single‐ and low‐copy RGAs were genetically mapped onto chromosomes 1H–7H (except 5H) using three barley double haploid (DH) mapping populations: Steptoe × Morex, Harrington × TR306 and LUGC × Bowman. Sequence analysis of the RGAs showed that they are members of a diverse group. As a result of BLAST searches, one RGA proved unique as it did not detect any significant hit. Another RGA is putatively functional, because it detected several barley expressed sequence tag (EST) matches. To physically map the RGAs, 13 sequences were used to screen a 6.3 × cv. ‘Morex’ bacterial artificial chromosome (BAC) library. After fingerprint analysis, eight contigs were constructed incorporating 62 BAC clones. These BAC contigs are of great value for positional cloning of disease resistance genes, because they span the regions where various barley R genes have been genetically mapped.     

Effectiveness and environmental stability of quantitative powdery mildew (Blumeria graminis) resistance among winter wheat cultivars

Powdery mildew in wheat (Blumeria graminis f. sp. tritici) is a major disease in Northern and Central Europe. The aim of the study was to analyse the effectiveness and environmental stability of quantitative powdery mildew resistance under high epidemic pressure in the field across years in the absence/presence of ineffective race‐specific resistances. Cultivars with and without Pm (major) genes were inoculated in three experiments with a genetically broad mildew population with all matching virulences. Resistance was measured three times by assessing the percentage of leaf area covered by powdery mildew on a plot basis (0–100%). Mean powdery mildew severity of the highly susceptible cv. ‘Kanzler’ varied across 10 years from 24% to 66% (Exp. 1). Means of three cultivars without Pm genes, ‘Ramiro’, ‘Miras’ and ‘Zentos’, and several cultivars with ineffective Pm genes varied quantitatively from 4% to 13%. Environmental stability of the quantitative resistances was on average higher than that of susceptible genotypes, as revealed by a regression approach. In the second experiment, all groups of cultivars with ineffective Pm gene(s) contained a large proportion of entries with a similar low mildew rating as the quantitatively resistant standard ‘Miras’. Mildew severity of pairs of cultivars with the same Pm gene(s) was significantly different across 6 years (Exp. 3) indicating the presence of additional quantitative resistances in some of these cultivars. In the analysis of variance, genotypic variance had a high impact (P < 0.01) with low importance of genotype × environment interaction. Consequently, heritabilites were high (0.95–0.97). In conclusion, breeders have already accumulated effective minor genes for powdery mildew resistance in many of the released German winter wheat cultivars. These quantitative resistances are long lasting, environmentally stable and provide a high level of protection to powdery mildew.     

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.     

Identification of genes for resistance to Puccinia striiformis f. sp. hordei in 18 barley genotypes

Barley genotypes Hor 1428, Hor 2926, Hor 3209, BBA 2890, Abyssinian 14, Grannelose Zweizeilige, and Stauffers Obersulzer are resistant to all races of Puccinia striiformis f. sp. hordei so far detected in the U.S.A. Heils Franken, Cambrinus, Astrix, Emir, Hiproly, Varunda, Trumpf,Mazurka, Bigo, BBA 2890, and I 5 are resistant to some races and susceptible to others. Previous studies showed that Hor 1428, Hor 2926, Hor 3209, Abyssinian 14, Stauffers Obersulzer, I 5, Heils Franken, Emir, Astrix, Hiproly, Varunda, and Trumpf each have two genes, and BBA 2890, Grannelose Zweizeilige, Cambrinus, Mazurka, Bigo, and BBA 809 each have a single genefor resistance. To determine the genes in specific genotypes and their relationships, all possible crosses were made among the 18 genotypes. Seedlings of parents and F₂ progeny were tested under controlledconditions for resistance to selected races that were avirulent on both parents. Based on segregation within the individual crosses to selected races, at least 26 of 30 genes detected in the 18 genotypes were different. Allelic and linkage relationships of some of the genes were determined. The genetic information should be useful for understanding the host-pathogen interactions and for control of stripe rust using resistance.     

The effect of specific and non-specific resistance in mixtures of barley or genotypes on infection by mildew (Erysiphe graminis f. sp. hordei) and on yield

Summary Preliminary field trials indicated that the yields of mixtures of barley cultivars partially resistant to mildew (Erysiphe graminis f. sp. hordei) were greater than those expected from the mean of their components. Subsequent trials in 1988, 1989 and 1990 demonstrated that such mixtures gave enhanced yield. However, fungicide treatment of mixtures gave significant positive and negative effects on yield. In mixtures with cultivars containing partial resistance there was evidence of a positive relationship between high yield loss due to mildew (low disease tolerance) in cultivars grown in monoculture and their yield advantage in mixtures. The converse resulted in yield disadvantage.     

Identification of resistance gene analogue markers closely linked to wheat powdery mildew resistance gene Pm31

Specific oligonucleotide primers, designed for the sequences of known plant disease resistance genes, were used to amplify resistance gene analogues (RGAs) from wheat genomic DNA. This method was applied in a bulked segregant analysis to screen for the RGA markers linked to the powdery mildew resistance gene Pm31, introgressed into common wheat from wild emmer. Two RGA markers (RGA200 and RGA390) were found to be closely linked to Pm31 and completely co‐segregating with the marker allele of Xpsp3029 linked to Pm31, with a genetic distance of 0.6 cM. These two RGA markers were then integrated into the formerly established microsatellite map of Pm31 region. The result showed the effectiveness of the RGA approach for developing molecular markers linked to disease resistance genes and demonstrated the efficiency of denaturing polyacrylamide‐gel electrophoresis for detecting polymerase chain reaction polymorphism.     

Identification of isolate specific sources of scald resistance in Turkish barley (Hordeum vulgare) accessions

Disease reactions to specific Rhynchosporium secalis isolates from western Canada were characterized on forty Turkish Hordeum vulgare accessions. One accession, MEH151-1, exhibited resistance against isolate WRS1860 and was also resistant to isolates WRS1493 and WRS1824 which were avirulent to the Rh gene. A related line, MEH151-2 exhibited resistance to WRS1493 but not to WRS1860. Crosses between both MEH151-1 and MEH151-2, and the susceptible cultivar, Harrington, were used to demonstrate a linkage between resistance to WRS1493 and an allele specific amplicon, Falc666. Resistance to WRS1860 imparted by MEH151-1 was mapped to the other side of the Falc666 locus. Falc666 was previously shown to be located near the centromere on the long arm of chromosome 3. Characterization of the genetic basis for the scald resistance phenotype exhibited by these Turkish accessions, coupled with the identification of marker linkages, provides evidence for genetic variation in scald resistance in this chromosomal region. This revised version was published online in July 2006 with corrections to the Cover Date.     

Differences in partial resistance of barley to powdery mildew (Erysiphe graminis DC F.SP. hordei Marchal) after chemomutagenesis. II. Reaction of mutants to pathotypes

Summary Seedlings of 45 barley mutants with partial resistance in the field and their parental cultivars Asse, Bomi, and Vada were exposed to six local and two foreign races of powdery mildew, in climatically controlled cabinets. Infectability, pustule size, infection grade, and infection type were estimated. No mutant did demonstrate strong race-specific reactions although some race-specific interactions of moderate grade could be detected. The results of infection of mutants with single races confirmed the quantitative character of their change in resistance determined in the earlier field assessments. The 20 investigated mutants with increased resistance expressed a lower level of disease with each of the used races. Out of the 10 mutants with higher susceptibility in the field, 4 mutants again exhibited higher degrees of infection over all the different races, while the remaining 6 mutants were not distinguishable from the original cultivar under the given growing conditions. Out of the tested 14 developmentally resistant mutants, only the 5 genotypes with different seedling reaction could be analyzed for race-specificity in this study. In all the above cases as the result of one gene mutation, quantitative shifts in level of infection were recorded over all the 8 races.Qualitative estimates of infection type were supported by data on the frequency of chlorotic or necrotic lesions and green islands on the infected leaves. But quantitative methods of assessment, e.g. infection frequency and pustule size, were more effective in studying partial resistance of the mutants. Infection grade, estimated visually 14 days after inoculation, was in agreement with the quantitative parameters accurately measured 7 days after inoculation. Therefore, its careful use can be recommended to speed the screening by narrowing down the materials. Differences in virulence level of the races were observed and their influence on race-specificity studies was discussed.     

The evaluation of adult plant resistance to powdery mildew (Erysiphe graminis f.sp. avenae) in transgressive lines of oats

Summary Previously identified segregant lines of oats with levels of adult plant resistance to powdery mildew (Erysiphe graminis f.sp. avenae) better than the resistant parent maintained this high level of resistance in field nurseries over two growing seasons. This enhancement was not expressed on inoculated detached leaves under laboratory conditions where no differences between the more resistant parent cv. Maldwyn and the most resistant segregants was detected. Reduced resistance was not detected in tests where leaf segments of the transgressive lines were inoculated with trained isolates. Problems associated with selecting for adult plant resistance under both field and laboratory conditions are discussed.     

Mapping of the apple powdery mildew resistance gene Pl1 and its genetic association with an NBS-LRR candidate resistance gene

Molecular markers for the major apple powdery mildew resistance gene Pl1 were identified and are presently used in marker-assisted selection in apple breeding. However, the precise map position of the Pl1 gene in the apple genome was not known. The objectives of this investigation were the identification of the Malus linkage group (LG) carrying the Pl1 locus, mapping of the resistance gene by simple sequence repeat (SSR) markers, and the analysis of genetic associations between the Pl1 gene and the numerous NBS-LRR resistance gene candidates already mapped in the apple genome. A two-step linkage mapping was used, based on two different apple families. The identification of LG 12 carrying Pl1 was performed indirectly by mapping the SCAR marker AT20 in an apple progeny for which there was a core genetic map but no mildew data available. Then, the position of Pl1 on LG 12 was determined by SSR markers in a second population which has been scored for mildew over 6 years in a greenhouse and in the field. The SSR Hi07f01, previously mapped on LG 12 [Tree Genet. Genomes, 2 (2006), 202] cosegregated with AT20 and was closely linked (∼1 cM) to the Pl1 gene. The TIR-NBS-LRR resistance gene analogue 15G11 mapped by the SSCP technique was also closely linked to the Pl1 resistance locus and might be a candidate for Pl1 itself, a second powdery mildew major resistance gene ( Pld , [Theor. Appl. Genet., 110 (2004), 175]), or two scab resistance genes ( Vg , [IOBC/WPRS Bull., 23 (2000), 245]; Vb , [Genome, 49 (2006), 1238]) which all seem to be located in a common R gene cluster at the distal end of apple LG 12.