Seasonal changes in pathotype complexity in French populations of barley powdery mildew

Airborne conidia of Erysiphe graminis f.sp. hordei were sampled in three regions and a single locality in the northern part of France for 2 years. Sampling was carried out in early spring, in late spring and in autumn, in order to separate the effects of winter barley cultivars, carrying few specific resistance alleles, and of spring barley cultivars, carrying diverse resistance alleles, on the structure of the pathogen population. Although complex pathotypes with three to 10 virulences were selected by spring cultivars, simple pathotypes, including a pathotype with the single unnecessary virulence allele Va22 , which formed a clear majority of the samples, remained dominant in early spring, when winter but not spring cultivars were growing. In early spring, simple pathotypes were more prevalent in the north, where the winter cultivars represented 90% of the barley acreage, than in the east, where winter cultivars represented 65%. In the west, the frequency of simple pathotypes was limited compared to the north, possibly because of the resistance allele Mlg in winter cultivars. The high frequency of simple pathotypes in early spring could be explained by a differential adaptation between simple and complex pathotypes or by delayed epidemics on spring cultivars compared to winter cultivars.     

Selection on Erysiphe graminis in pure and mixed stands of barley

The response of populations of Erysiphe graminis f. sp. hordei to selection by pure and mixed stands of three spring barley cultivars was studied in two field trials. The range of virulence of the pathogen genotypes selected in mixed host stands was dependent on the relative fitness of each genotype over all hosts. Unnecessary virulences were rapidly selected against on some hosts, but were less deleterious or favoured on others. In general there was selection for widely adapted pathogen genotypes in mixed host populations but this selection for flexibility limited the abilitytions in the absolute size of the pathogen population in host mixtures reduced the absolute frequencies of pathogen genotypes with combined virulences in comparison with those in pure stands. It is argued that host mixtures are therefore unlikely to favour rapid pathogen evolution towards races which are both widely adapted and highly virulent on all component cultivars which they can infect his definition docs not conform with conventional usage in population genetics.     

Effect of wheat cultivar mixtures on populations of Puccinia striiformis races*

This study quantifies the frequency of simple and complex races (races that can infect two or more components) of Puccinia striiformis in mixtures of wheat cultivars possessing different race-specific resistance genes. Treatments were designed so that the complex race changed depending on the host mixture, thus enabling us to observe the influence of pathogen complexity in different genetic backgrounds. Six cultivar mixtures and one pure stand of winter wheat were inoculated with three races of P. striiformis at two locations for two seasons. Potted plants of three winter wheat cultivars, each susceptible to one of the three races of the pathogen, were used to sample the pathogen during the field epidemics. Disease incidence on the differential cultivars was used to calculate the proportion of the three races in each treatment. The specific cultivars included in the mixtures influenced the frequencies of the three races. Increasing the number of virulent races in a mixture reduced the frequency of the complex race relative to the other two races. The results suggest that genetic background of the pathogen race, host composition, and interaction among pathogen races may be as important as cost of virulence in determining race frequencies in mixtures.     

Dynamics of the response of barley mildew to the use of sterol synthesis inhibitors1

Since 1980, the increase in the use of triazole fungicides to control Erysiphe graminis on barley in the UK has been accompanied by an increase in the frequency of pathogen isolates with different levels of reduced sensitivity to the fungicides. In each year, the rate of change varied, probably in relation to the degree of selection at different times in the host life cycle. In the first years, isolates with reduced sensitivity were less pathogenic than sensitive strains on a range of host cultivars. These differences were less obvious on cultivars with the Mla6 and Mlv resistance genes. More recently, reduced sensitivity has become widespread in the pathogen population and the general pathogenicity of this part of the population has increased. As a result, the yield benefit previously associated with mildew control by the triazoles is becoming severely diminished. This effect is not due primarily to the increase of highly insensitive isolates, but to the high frequency of isolates with intermediate insensitivity that are able to cause epidemic increase of the pathogen soon after fungicide application.     

Resistance in Australian barley (Hordeum vulgare) germplasm to the exotic pathogen Puccinia striiformis f. sp. hordei,causal agent of stripe rust

Eighty‐eight Australian and 10 international barley cultivars were assessed for resistance to the barley stripe (yellow) rust pathogen, Puccinia striiformis f. sp. hordei (Psh). All cultivars were tested for seedling resistance to two UK‐derived isolates of Psh (11.01 and 83.39) that were shown to differ in virulence based on responses on 16 differential barley genotypes. The 98 barley cultivars differed substantially in stripe rust response; 45% were susceptible to Psh 11.01, 53% to Psh 83.39 and 44% to both isolates. The observed diverse infection types (ITs) suggest the presence of both known and uncharacterized resistance. However, further multipathotype tests are required for accurate gene postulation. The Yerong × Franklin (Y×F) doubled haploid (DH) population was phenotypically assessed as seedlings using both Psh isolates. Yerong and Franklin were immune and highly resistant, respectively, to both isolates used in this study. Marker‐trait and QTL mapping identified a major effect on the long arm of chromosome 7H contributed by Franklin in response to all isolates. The resistance of Yerong was mapped to 113·96 and 169·38 cM on chromosome 5HL in response to Psh 11.01 and 83.39, respectively. The Psh resistance sources identified in this study can be used for further genetic analysis and introgression for varietal improvement.     

四川西南部小麦白粉菌群体毒性及遗传多样性分析

为明确四川西南部白粉菌群体毒性及其遗传变异情况,本文将2011年采自四川西南部的小麦白粉病标样进行单孢子堆分离纯化,共获得48个白粉菌菌株,并将其按采集地划分为4个地理群体。利用28份已知抗白粉病基因材料测定了群体的毒性频率,并运用SRAP分子标记技术探讨了其遗传多样性。毒性测定结果表明,白粉菌群体毒性较强,群体间毒性结构存在差异。供试群体对Pm1、Pm3a、Pm3b、Pm3d、Pm5a、Pm6、Pm19的平均毒性频率达50%以上,而Pm13、Pm XBD、Pm5b、Pm2+6、Pm5+6抗性保持良好,平均毒性频率在10%以下。群体间毒性遗传距离与地理距离之间有一定的相关性,但未达到显著水平。用筛选出的10对SRAP引物共获得160个清晰、稳定的扩增位点,多态性频率为50.63%;白粉菌群体遗传多样度(h)、Shannon信息指数(I)分别为0.198 8、0.322 7,遗传变异主要源于群体内部。群体间基因流数值均在6.50以上,说明四川西南部白粉菌群体间菌源迁移频繁,基因交流较为充分。M antel Test分析表明SRAP标记解释的群体遗传多样性与地理距离、毒性多样性间的相关性未达到显著水平。     

小麦白粉病菌生理小种的鉴定与病菌毒性的监测

１９９１～１９９４年小麦白粉病菌生理小种鉴定的结果表明,１１,１５号小种这几年的优势小种,３１５号小种有上升的趋势,用毒性频率分析的方法对所获数据进行分析表明,Ｖ１,Ｖ３,Ｖ５及Ｖ８等毒性的当前白粉菌群体中已基本固定（毒性频率接近１００％）,小麦白粉菌对Ｐｍ４ａ的毒性频率呈较快的上升趋势,说明Ｐｍ４ａ基因的使用应该审慎,白粉菌对Ｐｍ２１为目前最有效的抗白粉病基因,对北京,西南及江浙三地区小区白粉菌     

Quantitative evolution of aggressiveness of powdery mildew under two-cultivar barley mixtures

The effects of cultivar mixtures on the evolution of aggressiveness of barley powdery mildew ( Blumeria graminis f.sp . hordei ) were modelled. It was found that the rate and direction of evolution of pathogen aggressiveness in a race-non-specific system and value at equilibrium, depends on initial resistance levels, proportions of component cultivars, autodeposition rates, the relative magnitude of the benefit of autoinfection, and the cost of alloinfection of spores. In the model, mixing cultivars in any proportions tended to reduce the aggressiveness of pathogens at equilibrium compared with pure stands, but this effect decreased when two mixture components were extremely unbalanced in proportion. Under low and medium autodeposition rates, the best control of the evolution of the pathogen was achieved by mixing two components in roughly equal proportions. The magnitude of aggressiveness at equilibrium increased as autodeposition rates increased. Though the level of initial resistance of mixture components did not have an impact on the aggressiveness of pathogens at equilibrium, it strongly influenced the transient values of aggressiveness and therefore the total amount of disease caused over an evolutionary period. The cost to the pathogen of alloinfection and benefit of autoinfection per se did not affect the final level of aggressiveness, but did affect the time to reach equilibrium. However, the ratio of the cost to the benefit did influence the final aggressiveness of the pathogen.     

Disease severity and pathotype dynamics of Puccinia striiformis f.sp. tritici in Denmark

Disease severity of wheat yellow rust, Puccinia striiformis f.sp. tritici , was analysed in Denmark from 1985 to 1999 in relation to the effects of weather on winter survival, distribution of host cultivars and pathotype dynamics. Below-average temperatures in January and February (midwinter) reduced yellow rust on the susceptible cv. Anja, and in three of four growth seasons following cold winters no yellow rust was observed on any cultivar under natural conditions. The agronomic consequences of dispersal of yellow rust urediniospores from external sources to Denmark, in a period during which large areas were planted with relatively few wheat cultivars, were demonstrated in several cases, most evidently when the Yr9 and Yr17 resistance genes became ineffective. Yr9 was overcome by the pathogen in a period with severe yellow rust epidemics on commercial cultivars, while virulence for Yr17 was first observed in a year with almost no yellow rust. In contrast, the resistance in cv. Kraka ( Yr1, CV ) was increasingly effective in controlling yellow rust, because pathotypes with the matching combination of virulence declined in the pathogen population. Pathotype frequency dynamics were thus influenced by selection forces within the country, and by selection forces in areas where spores were spread to Denmark from outside. The importance of a sufficient level of partial resistance in the wheat germplasm to prevent too much damage by yellow rust epidemics, in the event that the resistance genes are overcome by the pathogen population, is emphasized.     

Measures of durability of resistance

ABSTRACT Conventional models for the durability of resistant cultivars focus on the dynamics of the frequency of resistance genes. This leads to a definition of the durability of resistance as the time from introduction of the cultivar to the time when the frequency of the virulence gene reaches a preset threshold. It is questionable whether this is the most appropriate way to measure durability. Here we use a simple epidemiological model to link population dynamics and population genetics to compare three measures of durability: (i) the expected time until invasion of the virulent genotype, by mutation or immigration, and subsequent establishment of a population (T(invasion)); (ii) the virulence frequency related measure of the time for the virulent genotype to take-over the pathogen population ( T(take-over)); and (iii) the additional yield, measured by the additional number of uninfected host growth days (T(additional)). Specifically, we show how the measures of durability are affected by deployment and epidemiological parameters. We use a combination of numerical solution and analytical approximation of a model for the population dynamics of avirulent and virulent genotypes of a pathogen growing in dynamically changing populations of resistant and susceptible cultivars. The three measures of durability are compared. Some consequences of the results for durable resistance in multilines and mixtures and the regional deployment of resistant cultivars are discussed.     

Analysis of diversity in populations of plant pathogens: the barley powdery mildew pathogen across Europe

Understanding population genetics and evolution within species requires recognition of variation within and between populations and the ability to distinguish between the potential causes of an observed distribution of variation. For this aim several established indices of diversity, and a novel one, were applied to population samples of the barley powdery mildew pathogen,Erysiphe graminis f. sp.hordei. Random spore samples were obtained from the air along transects through regions of interest across large parts of Europe in 1990. Significant geographical differences in diversity of virulence genotypes occurred among regional sub-samples. Diversity was highest in the samples from eastern Germany, Denmark and Austria, whereas the lowest values were found in the samples from Italy, southern France and parts of western Germany. Diversity in the pathogen population was generally related to the degree of diversification of host resistance in time and space, although there was considerable variation in ranking among different measures of diversity. Sensitivity to sample size proved to be the major problem with the use of several established indices of diversity. Working with very large sample sizes we used multiple random subsamples of various smaller sizes to determine how the mean index values changed with changing sample size. The Shannon index proved to be considerably affected by sample size, in contrast to the Simpson index that was therefore used as a global measure of diversity. Limits of confidence were estimated for the Simpson index using the bootstrap method of numerical resampling. The two aspects contributing to global diversity, richness and evenness, were considered separately to allow meaningful interpretation of the Simpson index. Random sub-sampling was used to reduce the influence of sample size differences for these measures. Dissimilarity, a novel measure of diversity for use in plant pathology, indicates the average number of major genes for host resistance against which pathotypes in a sample respond differently. It is thus able to account for the genetic relationship among pathotypes, which is not considered by any other index. The approaches developed in this study help to compare major forces driving evolution of large-scale populations of the barley mildew pathogen.     

Genetics of Virulence in Cochliobolus sativus and Resistance in Barley

ABSTRACT Spot blotch, caused by Cochliobolus sativus, is one of the most common foliar diseases of barley in the upper midwest region of the United States. To examine the genetics of host-specific virulence in C. sativus, a cross was made between isolate ND90Pr (which exhibits high virulence on barley genotype Bowman and low virulence on genotype ND 5883) and ND93-1 (which exhibits low virulence on both genotypes). Ascospore progeny segregated 48:55 for low virulence/high virulence on Bowman, indicating the presence of a single virulence gene in isolate ND90Pr. To complement the study of host-specific virulence in the pathogen, an experiment also was conducted on the genetics of specific resistance in the host. Progeny from a Bowman/ND 5883 cross were evaluated for their infection responses (IRs) to isolate ND90Pr at the seedling stage. The F(2) population segregated 1:3 for low IRs (resistant)/high IRs (susceptible), indicating the presence of a single resistance gene in genotype ND 5883. This result was confirmed in the F(3) generation, as a 1:2:1 ratio was found for homozygous resistant, segregating, and homozygous susceptible families, respectively. The data from this study demonstrate that both virulence in the pathogen and resistance in the host are under monogenic control in this specific host genotype/fungal isolate combination.     

江西省稻瘟病菌毒性的群体结构分析

用江苏省的吴江、赣榆、通州、高邮和宜兴等5个代表地区的324个稻瘟病菌株接种13个日本已知抗病基因品种，结果表明，江苏省稻瘟病菌对携带有Pi-Ka,Pi-ta,Pi-ta2和Pi-sh抗病基因品种的可致病比例较高，毒力频率达64．2％－97．2％，对携带有Pi-iPi-z和Pi-b抗病基因的品种毒力频率较低，在0－11．9％之间；江苏省稻瘟病菌毒性的群体结构年度间有一定变化，表现为对某些抗病基因品种毒力频率的明显上升或下降。从江苏省10年主栽品种上分离的稻瘟病菌株对13个已知抗病基因品种的毒力频率测定结果表明，不同类型主栽品种上分离的菌株其毒性结构组成有一定差异,表明江苏省水稻主栽品种的抗性类型存在一定差异。     

A comparative assessment of potential components of partial disease resistance to <Emphasis Type="Italic">Fusarium</Emphasis> head blight using a detached leaf assay of wheat,barley and oats

The relative resistance of 15 winter barley, three winter wheat and three winter oat cultivars on the UK recommended list 2003 and two spring wheat cultivars on the Irish 2003 recommended list were evaluated using Microdochium nivale in detached leaf assays to further understand components of partial disease resistance (PDR) and Fusarium head blight (FHB) resistance across cereal species. Barley cultivars showed incubation periods comparable to, and latent periods longer than the most FHB resistant Irish and UK wheat cultivars evaluated. In addition, lesions on barley differed from those on wheat as they were not visibly chlorotic when placed over a light box until sporulation occurred, in contrast to wheat cultivars where chlorosis of the infected area occurred when lesions first developed. The pattern of delayed chlorosis of the infected leaf tissue and longer latent periods indicate that resistances are expressed in barley after the incubation period is observed, and that these temporarily arrest the development of mycelium and sporulation. Incubation periods were longer for oats compared to barley or wheat cultivars. However, oat cultivars differed from both wheat and barley in that mycelial growth was observed before obvious tissue damage was detected under macroscopic examination, indicating tolerance of infection rather than inhibition of pathogen development, and morphology of sporodochia differed, appearing less well developed and being much less abundant. Longer latent periods have previously been related to greater FHB resistance in wheat. The present results suggest the longer latent periods of barley and oat cultivars, than wheat, are likely to play a role in overall FHB resistance if under the same genetic control as PDR components expressed in the head. However the limited range of incubation and latent periods observed within barley and oat cultivars evaluated was in contrast with wheat where incubation and latent periods were shorter and more variable among genotypes. The significance of the various combinations of PDR components detected in the detached leaf assay as components of FHB resistance in each crop requires further investigation, particularly with regard to the apparent tolerance of infection in oats and necrosis in barley, after the incubation period is observed, associated with retardation of mycelial growth and sporulation.     

Application of real-time and multiplex polymerase chain reaction assays to study leaf blotch epidemics in barley

ABSTRACT Leaf blotch, caused by Rhynchosporium secalis, was studied in a range of winter barley cultivars using a combination of traditional plant pathological techniques and newly developed multiplex and real-time polymerase chain reaction (PCR) assays. Using PCR, symptomless leaf blotch colonization was shown to occur throughout the growing season in the resistant winter barley cv. Leonie. The dynamics of colonization throughout the growing season were similar in both Leonie and Vertige, a susceptible cultivar. However, pathogen DNA levels were approximately 10-fold higher in the susceptible cultivar, which expressed symptoms throughout the growing season. Visual assessments and PCR also were used to determine levels of R. secalis colonization and infection in samples from a field experiment used to test a range of winter barley cultivars with different levels of leaf blotch resistance. The correlation between the PCR and visual assessment data was better at higher infection levels (R(2) = 0.81 for leaf samples with >0.3% disease). Although resistance ratings did not correlate well with levels of disease for all cultivars tested, low levels of infection were observed in the cultivar with the highest resistance rating and high levels of infection in the cultivar with the lowest resistance rating.     

Genotypic diversity in barley powdery mildew populations in northern France

Diversity in populations of Erysiphe graminis ( Blumeria graminis ) f.sp. hordei was studied with virulence and molecular markers. Isolates were sampled in two locations in northern France from a winter barley cultivar (Plaisant) and a spring cultivar (Caruso). Only a few pathotypes (determined by virulence markers) were common. The rest of the population was diverse. Diversity within common pathotypes, estimated by five RAPD and two SCAR markers, was generally high, except for one pathotype, which was frequent on Plaisant. This pathotype carried only one virulence, Va22, out of the 11 virulences tested. It appeared as a clonal lineage, which had occurred previously, at least in 1992, in northern France, demonstrating survival of asexual lineages in populations that often reproduce sexually.     

Distribution of pathotypes with regard to host cultivars in French wheat leaf rust populations

ABSTRACT Isolates of wheat leaf rust collected from durum and bread wheat cultivars in France during 1999-2002 were analyzed for virulence on 18 Thatcher lines with single genes for leaf rust resistance (Lr genes). Sampling focused on the five most widely grown bread wheat cultivars (two susceptible and three resistant) to allow statistical comparison of diversity indexes between the cultivars. Leaf rust populations from durum and bread wheats were different. The diversity of the bread wheat leaf rust pathotypes, as measured by the Shannon index, ranged from 2.43 to 2.76 over the 4 years. Diversity for wheat leaf rust resistance was limited in the host since we postulated only seven seedling resistance genes in the 35 cultivars most widely grown during 1999-2002. Leaf rust populations were strongly differentiated for virulence within bread wheat cultivars, and diversity was higher on those that were resistant, mainly due to a more even distribution of virulence phenotypes than on susceptible cultivars. The pathogen population on the susceptible cv. Soissons was largely dominated by a single pathotype (073100), whereas all other pathotypes virulent on cv. Soissons either decreased in frequency or remained at a low frequency during the period studied. Several pathotypes including the most complex one were found only on resistant cultivars, even though most of them were virulent on the susceptible cv. Soissons. Specific interactions were necessary, but not always sufficient, to account for pathotype distribution and frequencies on the cultivars, suggesting that selection for virulence to host resistance genes is balanced by other selective forces including selection for aggressiveness.     

Genetic structure of a Pyrenophora teres f. teres population over time in an Australian barley field as revealed by Diversity Arrays Technology markers

Net form of net blotch caused by Pyrenophora teres f. teres (Ptt) is a major foliar disease of barley (Hordeum vulgare) worldwide. Knowledge of the evolution of Ptt pathogen populations is important for development of durable host-plant resistance. This study was conducted to investigate changes in genetic structure of a Ptt population within a barley field during three cropping years. The susceptible barley cultivar Henley was inoculated with Ptt isolate NB050. Leaf samples were collected during the years 2013–15 and 174 single spore Ptt isolates stored. Genotyping using Diversity Arrays Technology markers identified that 25% of isolates were clones of the inoculated isolate and 75% of isolates were multilocus genotypes (MLGs) differing from the original inoculated genotype. The novel genotypes probably originated from a combination of windborne spores from neighbouring fields, infected seed and sexual recombination in the field. The rapid change in the genotypic composition of the Ptt population in this study suggests adaptive potential of novel genotypes and demonstrates the need for barley breeders to use multiple sources of host-plant resistance to safeguard against resistance being overcome.     

大麦种质对叶斑病的抗性鉴定与评价

 由麦根腐平脐蠕胞菌引起的叶斑病在世界各大麦种植区均有发生,严重影响大麦的产量和品质。选育和应用抗性品种是防控该病害最有效的策略,然而可利用的抗源非常有限。在本研究中对中国233份具有代表性的大麦种质资源进行成株期抗叶斑病田间人工接种鉴定,发现只有垦啤麦5号等10份材料对3个供试菌株都表现抗病,仅占供试材料的4.3%。另外对37份国内外重要的叶斑病抗源材料进行苗期及成株期抗叶斑病鉴定,结果显示成株期抗叶斑病材料所占比例为41%~46%,苗期抗性材料所占比例为50%~64%,其中ND17293等11份材料在苗期和成株期对3个菌株均表现为抗病,可作为抗源继续加以利用;基于上述鉴定结果,进一步分析发现供试大麦苗期对三个菌株的抗病比例均高于成株期抗病比例,说明大麦在不同生育期对叶斑病的抗性存在较大差异。另外发现大麦对B. sorokiniana不同致病类型的抗性也存在明显的专化性。     

Survey of benomyl resistance in Pseudocercosporella herpotrichoides on winter wheat and barley in England and Wales in 1983

Surveys of 528 randomly selected commercial fields in England and Wales in 1983 showed that isolates of Pseudocercosporella herpotrichoides resistant to benomyl were common and widespread on winter wheat and winter barley. During June and July resistant strains were recovered from 16 to 66% of fields and the proportion of resistant isolates was 37–52%. The frequency of occurrence of resistant isolates was positively correlated with the number of applications of carbendazimgenerating (MBC) fungicides during the previous 8 years. Benomyl resistance was found in 21–48% of fields where it was claimed that MBC had never been used, and 7–28% of isolates from those fields, were resistant. Resistance was more frequent in R-type than in W-type isolates of the pathogen. The proportion of R-type isolates decreased as the number of previous winter wheat crops rose and increased with the number of winter barley crops grown and with the number of MBC sprays applied to crops in previous years. The implications of these findings for the control of eyespot in England and Wales are discussed.