Virulence Frequences of Puccinia triticina in Germany and the European Regions of the Russian Federation

From 2001 to 2003, leaf rust was collected in different regions of Germany and the Russian Federation to generate single spore isolates and to study the structure of the pathogen populations by analyses of virulence. The virulence of isolates was tested with 38 near‐isogenic lines each carrying a different resistance gene. The analyses of variance revealed significant effects for the frequency of virulent isolates, the regions and most interactions with years and regions, but no significance was found for the effects of years. In Germany, an increase of virulence frequencies was detected for Lr1 and Lr2a while a decrease was found for Lr3a, Lr3bg and Lr3ka. Such clear trends did not occur in Russia which may be due to the great agroclimatic differences between regions. The variance of the frequency of virulent isolates was used to estimate adequate sample sizes for the analysis of regional populations of leaf rust. This procedure resulted in more reliable information about the dynamic processes within the pathogen populations. In 2002 and 2003, all pathotypes in Germany had a combined virulence to Lr1, Lr2a, Lr2b, Lr15, Lr17 and Lr20 supplemented by a few other genes. The complexity of virulence was lower in the most frequent pathotypes. In Russia virulence to the alleles at locus Lr3 was very common. Using detached leaf segments in Germany and Russia it turned out that the most virulent pathotypes carry 34 and 32 virulence genes, respectively. Virulence to Lr9, Lr19, Lr24 and Lr38 was rare or even absent. The use of major genes, not overcome by corresponding virulent pathotypes, may contribute to more durable types of resistance in case they are combined with genes having different effects, e.g. adult plant resistance.     

Genetic analysis of resistance to blackspot (Diplocarpon rosae) in tetraploid roses

  Diplocarpon rosae is the causal agent of rose blackspot, one of the most severe diseases of field-grown roses. The genetics of resistance to this pathogen was investigated in crosses between tetraploid rose genotypes. The hybrid breeding line 91/100-5, which exhibits a broad resistance to all isolates tested so far, was selfed to produce an F₂ population, backcrossed to the susceptible tetraploid variety ‘Caramba’ and crossed to the susceptible varieties ‘Heckenzauber’, ‘Pariser Charme’ and ‘Elina’. Infection experiments resulted in segregation ratios consistent with the presence of a single dominant resistance locus in the duplex configuration in the hybrid 91/100-5. This suggests, together with previous data on the race structure of the fungus, a “gene-for-gene” type of interaction in the pathosystem Diplocarpon/Rosa. We propose to designate this gene Rdr1, which is the first resistance gene described in the genus Rosa. The advantages and limitations of such an interaction type for future rose breeding programmes and for marker-assisted selection strategies are discussed. Received: 29 August 1997 / Accepted: 19 September 1997     

HOST-PATHOGEN INTERACTIONS IN NATURAL POPULATIONS OF LINUM MARGINALE AND MELAMPSORA LINI: I. PATTERNS OF RESISTANCE AND RACIAL VARIATION IN A LARGE HOST POPULATION

Populations of wild flax, Linum marginale and its associated rust fungus Melampsora lini growing at Kiandra, New South Wales, Australia, were sampled during the 1986–1987 growing season. Thirteen different races of M. lini were detected in a sample of 96 isolates. The distribution of isolates was uneven: race A comprised 73% of the samples; race N, 8%; and race H, 5%; while the remaining races were represented by only one or two samples. The dominance of race A increased over the course of the growing season, comprising 67% of the early season samples and increasing to 78% for those collected late in the season. The overall diversity of the pathogen population decreased late in the growing season, but this trend was not statistically significant. The average virulence of individual isolates of the pathogen population increased during the growing season. This trend was most pronounced among the minor races, where the mean number of differential hosts infected increased from 4.58 for early season samples to 5.12 and 5.08 for mid and late season samples, respectively. In contrast to the virulence pattern in the pathogen, the L. marginale population displayed a more even distribution of resistance. In a sample of 67 plants 10 resistance phenotypes were detected from their pattern of resistance/susceptibility to seven pathogen isolates. No phenotype had a frequency that exceeded 30%. Resistance phenotypes were randomly distributed on both a population level and on a fine scale.     

Genetic analysis of resistance to yellow rust in hexaploid wheat using a mixture model for multiple crosses

DNA-based molecular markers have been used in numerous studies for tagging specific genes in wheat for subsequent use in marker-assisted selection. Usually in plant breeding, procedures for mapping genes are based on analysis of a single segregating population. However, breeding programmes routinely evaluate large numbers of progeny derived from multiple-related crosses with some parental lines shared. In most such related crosses, the number of progeny is quite small. Thus, statistical techniques for detecting quantitative trait loci (QTLs) using data from conventional multi-cross breeding programmes are interesting. The objective of this study is to present a mixture model for QTL mapping in crosses of multiple inbred varieties with non-normal phenotype distributions and to use this model to map QTLs for yellow rust resistance in elite wheat breeding material. Three doubled haploid populations consisting of 41, 42 and 55 lines, respectively, originating from four parental varieties were studied. Multi-cross QTL analysis with three specific pathogen isolates of Puccinia striiformis f. sp. tritici and a mixture of the isolates revealed QTLs for resistance at four different genomic locations. These QTLs were found on chromosome 2AL, 2AS, 2BL and 6BL and explained between 21 and 41% of the phenotypic variation. Two of these QTLs, one on the long arm of chromosome 2A and one on the short arm of chromosome 2A were identical to the known yellow rust resistance genes Yr32 and Yr17, respectively, whereas the QTLs located on the long arms of chromosomes 2B and 6B may reflect types of resistance to yellow rust, which have not previously been mapped.     

HOST-PATHOGEN INTERACTIONS IN NATURAL POPULATIONS OF LINUM MARGINALE AND MELAMPSORA LINI: II. LOCAL AND REGIONAL VARIATION IN PATTERNS OF RESISTANCE AND RACIAL STRUCTURE

Spatial variation in the resistance structure of Linum marginale (wild flax) populations to the rust fungus Melampsora lini, and in the racial structure of this pathogen, was investigated by sampling 10 populations distributed throughout the Kosciusko National Park, New South Wales, Australia. Considerable differences were found among populations in the structure of both host and pathogen. Host populations were divided into three broad categories: (1) populations susceptible to all testing races; (2) populations containing a strictly limited number of resistant phenotypes; and (3) populations with a considerable diversity of resistant phenotypes. The pathogen populations also showed a range of diversity. The major differences between these populations were determined by the occurrence and frequency of four common races of pathogen (races A, E, K, and N). These differences were apparent both at a regional spatial scale (over the 100 km separation of the most distant populations) and at a local scale where major differences were detected between two populations only 300 m apart. The distribution of the four common races of M. lini was consistent with the hypothesis that a fitness cost was associated with unnecessary virulence. In general, however, differences in the structure of pathogen populations from genetically very similar host populations implied that, in addition to host resistance genes, other evolutionary forces are also important in determining the genetic structure of individual pathogen populations.     

Incidence, importance and virulence of Puccinia hordei on barley in New Zealand

Brown rust of barley (Puccinia hordei) is widespread in New Zealand and causes crop losses in some years. The cultivars presently grown lack adequate genetic resistance but a highly resistant line has been produced. Twelve different virulence combinations were identified in the pathogen population using the Welsh differential lines; there were differences between North Island and South Island isolates; and no virulence was observed for resistance conferred by factors Pa 3 and Pa 7.     

The application of DNA marker and doubled-haploid technology for stacking multiple stem rust resistance genes in wheat

In wheat, the use of gene “pyramids” or “stacks” of major genes that confer resistance to all local strains of the fungal stem rust pathogen Puccinia graminis f. sp. tritici (Pgt) can increase durability of resistance where wheat cultivars with the single gene components are not widely deployed. Stacking two or more resistance genes becomes a breeding challenge, particularly when pathogen races that discriminate the genes are not available. The use of DNA markers and doubled-haploid technology provides a route for producing lines homozygous for multiple resistance genes. We have applied this approach to produce gene pyramids of two or more of the stem rust resistance genes Sr24 and new sources of SrR, Sr31 and Sr26 on reduced alien chromatin in the genetic backgrounds of Westonia and Pavon wheat. These genes, which are all derived from “alien” sources (SrR and Sr31 from rye, Sr24 and Sr26 from Agropyron elongatum) each provide resistance to all currently known pathotypes of Pgt in Australia, and SrR and Sr26 also provide resistance against all the variants of stem rust race Ug99 (TTKS group).     

Resistance toMelampsora larici-epitea leaf rust inSalix: analyses of quantitative trait loci

Quantitative resistance ofSalix toMelampsora larici-epitea leaf rust was studied in 2Salix mapping populations. One population was a backcross between aS. schwerinii ×S. viminalis hybrid andS. viminalis, and the other was an F₂ population betweenS. viminalis andS. dasyclados. A leaf disc bioassay was used to study the components of quantitative resistance (latent period, uredinia number, and uredinia size) to 3 isolates of the leaf rust. The analysis of quantitative trait loci (QTLs) revealed 9 genomic regions in the backcross population and 7 genomic regions in the F₂ population that were important for rust resistance, with QTLs explaining 8–26% of the phenotypic variation. An important genomic region was identified for the backcross population in linkage group 2, where QTLs were identified for all resistance components for 2 of the rust isolates. Four of the QTLs had overlapping mapping intervals, demonstrating a common genetic background for latent period, uredinia diameter, and uredinia number. QTLs specific to some rust isolates and to some resistance components were also found, indicating a combination of common and specific mechanisms involved in the various resistance components. Breeding implications in relation to these findings are discussed.     

Host-pathogen diversity in a wild system: Chondrilla juncea–Puccinia chondrillina

The resistance structure of a Turkish population of the clonal, apomictic composite Chondrilla juncea and the pathotypic structure of a co-occurring population of its obligate rust pathogen, Puccinia chondrillina, was determined by sequential inoculation of 19 host lines with 15 pathogen isolates each derived from single pustules collected from separate plants among the host population. The resultant matrix of resistant and susceptible reactions provides strong circumstantial evidence for a gene-for-gene interaction. Seven distinct pathotypes were detected in the pathogen population. One of these comprised 53% of the population, a second comprised 13%, while the remaining five pathotypes were each detected only once. The host population was similarly diverse, being composed of eight resistance phenotypes, only two of which were represented by more than one host line. Although C. juncea is apomictic, there was only 58% congruence between host resistance and multi-locus isozyme phenotype categories within this population. Pathotypic phenotypes of 13 other isolates of P. chondrillina collected from ten other Turkish and three more distant populations of C. juncea were markedly different from those found in the population studied in detail. There was no obvious relationship between the degree of geographic separation of pathotypes and their ability to attack particular C. juncea lines in this or three other populations represented by single host lines. Received: 10 March 1997 / Accepted: 4 August 1997     

Molecular mapping of genes for race-specific overall resistance to stripe rust in wheat cultivar Express

‘Express’, a hard red spring wheat cultivar that has been widely grown in the western United States, is used to differentiate races of Puccinia striiformis f. sp. tritici, the causal fungal pathogen of wheat stripe rust. To identify genes conferring race-specific, overall resistance to stripe rust, Express was crossed with ‘Avocet S’. The parents and F₁, F₂, F₃ and F₅ populations were tested with races PST-1, PST-21, PST-43, and PST-45 of P. striiformis f. sp. tritici in the seedling stage under controlled greenhouse conditions. Two dominant genes for resistance to stripe rust were identified, one conferring resistance to PST-1 and PST-21, and the other conferring resistance to all four races. Linkage groups were constructed for the resistance genes using 146 F₅ lines to establish resistance gene analog and chromosome-specific simple sequence repeat marker polymorphisms. The gene for resistance to races PST-1 and PST-21 was mapped on the long arm of chromosome 1B, and that conferring resistance to all four races was mapped on the long arm of chromosome 5B. We temporarily designate the gene on 1BL as YrExp1 and the gene on 5BL as YrExp2. Polymorphism of at least one of the two markers flanking YrExp2 was detected in 91% of the 44 tested wheat genotypes, suggesting that they would be useful in marker-assisted selection for combining the gene with other resistance genes into many other wheat cultivars. Knowledge of these genes will be useful to understand recent virulence changes in the pathogen populations.     

RESISTANCE AND VIRULENCE STRUCTURE IN TWO LINUM MARGINALE-MELAMPSORA LINI HOST-PATHOGEN METAPOPULATIONS WITH DIFFERENT MATING SYSTEMS

Different patterns of resistance to six pathotypes of Melampsora lini were detected in 11 populations of Linum marginale distributed across two metapopulations. The two metapopulations (mountains and plains of New South Wales, Australia) differed in the annual cycle of disease development, which barely overlapped, and in the growth cycle and mating system of the host. Host populations in the mountains metapopulation were highly inbred, whereas those on the plains showed appreciable levels of outcrossing. Within each metapopulation there was significant variation among component populations in (1) levels of host resistance to individual pathogen isolates; (2) mean levels of resistance to all six isolates; (3) the number of resistance phenotypes present and the evenness of their distribution within the population; and (4) the average number of pathogen lines to which individual hosts were resistant. A more limited comparison of pathogen populations from the two metapopulations (two from each) found greater similarities in the structure of populations and particular virulence frequencies within, rather than among, the two metapopulations. Differences in host outcrossing rates between the two metapopulations are reflected in marked differences in the overall level of resistance, its partitioning within and among populations, the number and distribution of resistance phenotypes in the two areas, and the level of polymorphism for specific virulence factors in the pathogen, with the plains metapopulation showing consistently higher values. However, these differences were not significant. In general, variation for all parameters was just as great among populations within a metapopulation as between the two metapopulations.     

Isolate-specific QTLs for partial resistance to Puccinia hordei in barley

By using a high-density AFLP marker linkage map, six QTLs for partial resistance to barley leaf rust (Puccinia hordei) isolate 1.2.1. have been identified in the RIL offspring of a cross between the partially resistant cultivar ’Vada’ and the susceptible line L94. Three QTLs were effective at the seedling stage, and five QTLs were effective at the adult plant stage. To study possible isolate specificity of the resistance, seedlings and adult plants of the 103 RILs from the cross L94×’Vada’ were also inoculated with another leaf rust isolate, isolate 24. In addition to the two QTLs that were effective against isolate 1.2.1. at the seedling stage, an additional QTL for seedling resistance to isolate 24 was identified on the long arm of chromosome 7. Of the eight detected QTLs effective at the adult plant stage, three were effective in both isolates and five were effective in only one of the two isolates. Only one QTL had a substantial effect at both the seedling and the adult plant stages. The expression of the other QTLs was developmental-stage specific. The isolate specificity of the QTLs supports the hypothesis of Parlevliet and Zadoks (1977) that partial resistance may be based on a minor-gene-for-minor-gene interaction. Received: 16 February 1999 / Accepted: 20 February 1999     

Virulence patterns of barley yellow rust and effective resistance genes to Puccinia striiformis in three parts of Iran

Stripe rust of barley (Hordeum vulgare L.), caused by Puccinia striiformis f. sp. hordei, is a serious problem of barley production in many parts of the world. Monitoring of the pathogen virulence factors and their changes provides basic information for development of an early warning system to breeders and researchers. To monitor the regular virulence changes, trap nurseries comprising of 12 barley differential sets were planted at different parts of Iran in six consecutive years 2007–2012. When the infection and severity under natural infection on susceptible cultivar Afzal as the check was high, then the response of each line was assessed using modified Cobbs scale. Results revealed that no virulence was observed on plants with resistance genes rpsEm1, rpsEm2, rpsHF, Rps4, rpsVa1, rpsVa2 and rpsAst. Therefore, these genes were considered effective genes and can be used to pyramid with those for race-non-specific resistance genes to achieve more durable and highly effective resistance to stripe rust. The plants with the resistance genes rps2, Rps1.b, Rps3 and rpsI5 showed susceptible reaction and virulence was observed on them, thus their resistance genes were considered ineffective.     

The genetics of rust resistance in sugar cane seedling populations

The inheritance of rust resistance was studied in sugar cane seedling populations using a factorial mating design over 1 summer and 2 winter seasons. Frequency distributions for rust infection pooled over 2 winter seasons for resistant x resistant parents were highly skewed with the majority of progenies grouped towards the resistant classes, whereas crosses between susceptible x highly susceptible parents tended to be skewed with the majority of progenies grouped towards the susceptible ones. Both categories of crosses produced transgressive segregants at either extremes. Distribution of infection within progeny of the selfed resistant parent ’R 570’ and distribution in the majority of crosses tended to support the hypothesis of a major gene with a dominant effect for resistance. However, the action of other minor genes acting in a quantitative way is also suggested. The female (F) and male (M) variance components were very important, and F×M interaction indicated the existence of non-additive genetic effects. F×S, M×S and F×M×S interaction mean squares were generally low or insignificant. Broad-sense heritability for the individual season ratings and for the combined ratings was high (0.75–0.90), whereas narrow-sense heritability was generally moderate (0.40–0.52) with the additive genetic effects accounting for 44–68% of the total genetic variation. The implications of these findings in the breeding for rust resistance in the local programme are discussed. Received: 2 June 1999 / Accepted: 22 June 1999     

Coevolution at multiple spatial scales: Linum marginale–Melampsora lini – from the individual to the species

Coevolutionary processes are intrinsically spatial as well as temporal, and occur at many different scales. These range from single populations dominated by demographic and genetic stochasticity, to metapopulations in which colonisation/extinction dynamics have a large influence, and larger geographic regions where phylogenetic patterns and historical events become important. We present data for the genetically and demographically well-characterised plant host–pathogen interaction, the Linum marginale–Melampsora lini system, and use this to demonstrate the varying nature of resistance and virulence structure across these spatial scales. At the within population level, our results indicate considerable variability in resistance and virulence, but little evidence of coordinated changes in host and pathogen. Studies involving comparisons among multiple demes within a single metapopulation show that adjacent populations often have asynchronous disease dynamics and large differences in diversity and frequency of resistance and virulence phenotypes. Nevertheless, at this scale, there is also evidence of spatial structure in that more closely adjacent host populations are significantly more likely to have similar resistance phenotypes and mean levels of resistance. At larger scales, comparisons among adjacent metapopulations indicate that quantitative differences in host mating system and other life history features can have further major consequences for how host and pathogen variation is packaged. Finally, comparisons at continental and among host-species levels show variation consistent with specialisation and speciation in the pathogen. This revised version was published online in July 2006 with corrections to the Cover Date.     

High diversity of genes for nonhost resistance of barley to heterologous rust fungi

Inheritance studies on the nonhost resistance of plants would normally require interspecific crosses that suffer from sterility and abnormal segregation. Therefore, we developed the barley-Puccinia rust model system to study, using forward genetics, the specificity, number, and diversity of genes involved in nonhost resistance. We developed two mapping populations by crossing the line SusPtrit, with exceptional susceptibility to heterologous rust species, with the immune barley cultivars Vada and Cebada Capa. These two mapping populations along with the Oregon Wolfe Barley population, which showed unexpected segregation for resistance to heterologous rusts, were phenotyped with four heterologous rust fungal species. Positions of QTL conferring nonhost resistance in the three mapping populations were compared using an integrated consensus map. The results confirmed that nonhost resistance in barley to heterologous rust species is controlled by QTL with different and overlapping specificities and by an occasional contribution of an R-gene for hypersensitivity. In each population, different sets of loci were implicated in resistance. Few genes were common between the populations, suggesting a high diversity of genes conferring nonhost resistance to heterologous pathogens. These loci were significantly associated with QTL for partial resistance to the pathogen Puccinia hordei and with defense-related genes.     

Pyramiding of alleles with different rust resistance specificities in <Emphasis Type="Italic">Linum usitatissimum</Emphasis> L.

Transgenic flax plants expressing flax rust resistance specificities conferred by L ² and L ¹⁰ alleles of L locus were produced through Agrobacterium tumefaciens-mediated transformation of hypocotyl segments or anther culture derived-calli. Transgenic plants were characterized by PCR amplification and Southern hybridization. Homozygous transgenic lines containing a single locus of effective transgene were isolated by consecutive progeny analyses of transgenic plants. In addition, homozygous lines were directly obtained from transformation of haploid cells followed by spontaneous or artificial chromosome doubling when anther culture derived-calli were used as the explants. All transgenic plants containing L ² transgene expressed L ² flax rust resistance specificity and had unambiguous infection type (IT) “0” (immune) reactions to flax rust race 22, which is virulent to endogenous L ⁶ and K ¹ genes present in the untransformed Linola™1084. Transgenic plants containing L ¹⁰ transgene exhibited various levels of resistance to flax rust race 191. Five plants had IT “fleck” (immune) reactions, similar to the reactions with the L ¹⁰ rust differential line, Bolley Golden Selection. The enhanced resistance to rust race 191 in these transgenic plants was attributed to the expression of L ¹⁰ rust resistance specificity. Further evaluation of the transgenic plants containing L ² or L ¹⁰ transgene to flax rust race 258 and race 247 respectively showed that the endogenous rust resistance specificities were not modified. The implication of this study in producing transgenic flax plants with multiple resistance specificities and for crop improvement using molecular breeding strategy is discussed.     

Evolution of pathogenicity in Plasmopara halstedii (sunflower downy mildew)

Comprehension of the processes of co-evolution between the pathogen and its host plant is very important, particularly in the case of obligate pathogen as Plasmopara halstedii which cannot develop only on sunflower. The influence of selection pressure exercised by qualitative resistance in sunflower plants on evolution of pathogenicity was analysed in pathogenic populations of P. halstedii. This selection pressure led a new virulence to appear in P. halstedii isolates carrying several levels of aggressiveness. It seems that the qualitative resistance selection pressure plays an important role in the evolution of this pathogen, and these changes on the level of pathogenicity may help to a better adaptation of P. halstedii in the presence of intensive use of qualitative resistance.     

Effect of chronic irradiation on plant resistance to biotic stress in 30-km chernobyl nuclear power plant exclusion zone

It was established in greenhouse experiments that infection with powdery moldew (Erysiphe graminis DC. f. sp. tritici Em. Marchal) and brown rust (Puccinia triticana Erikss. & Henn.) of three wheat (Triticum aestivum L.) cultivars (Mironovskaya 808, Polesskay 70, and Kiyanka) grown from seeds, collected in the Chernobyl exclusion zone, was 1.5–2.0 times higher than of plants grown from control seeds. On field trials in the Chernobyl zone, wheat plant resistance to biotic stress was reduced. At artificial infection with brown rust, the disease development was enhanced on plots with increased radiation background. One of the mechanisms of the declined phytoimmunity potential under the action of low doses of chronic irradiation is evidently a reduced activity of plant proteinase inhibitors. Thus, in wheat and rye (Secale cereale L., cv. Saratovskaya) grains, their activity reduced by 35–60% as compared to control. Active form and race formation in the population of the cereal stem rust causal agent (Puccinia graminis Pers.) was observed in the Chernobyl zone. A “new” population of this fungus with high frequency of more virulent clones than in other Ukraine regions was distinguished. The results obtained independently in greenhouse and field trials performed in the Chernobyl zone demonstrated radiation stress influence on the pathogen-plant interactions. They indicate a necessity of monitoring the microevolutionary processes occurring in both plants and their pathogens under conditions of technogenic stresses.     

Phenotypic and genetic patterns of resistance to the pathogen Phakopsora pachyrhizi in populations of Glycine canescens

Summary Phenotypic patterns of resistance to nine races of the pathogen Phakopsora pachyrhizi (soybean rust) in two natural populations of Glycine canescens were determined. In both populations there was considerable variability both within and between different host lines in their resistance or susceptibility to the nine different pathogen races. The genetic basis of these patterns of resistance was analyzed through an extensive series of crosses. In both host populations resistance was conditioned by single dominant genes with major phenotypic effects. One, two or three such genes were present in each host line. Using the principles of the gene-for-gene hypothesis, knowledge about the number of resistance genes present in each host line and by cross comparison of the phenotypic patterns of disease resistance detected in each line, estimates were made of the number of resistance genes or alleles present in each population of G. canescens. The two populations contained a minimum of 10 and 12 resistance genes. The relevance of these results to agriculture is discussed briefly.