Levels of resistance of Erysiphe graminis f.sp. hordei to the systemic fungicide triadimenol

A random sample of conidiospores ofErysiphe graminis f.sp.hordei was obtained from the atmosphere above Cambridge, England, by incubating them on barley seedlings exposed on three dates in 1985. The asexual progeny of each spore was tested for its response to a range of doses of a systemic triazole fungicide, triademenol. Principal components analysis of the data revealed that the majority of isolates had a distinct level of response to triadimenol, being resistant to triadimenol at the rate apoplied to seed commercially. The remaining isolates appeared either to be sensitive to the lowest dose of triadimenol used, or to have one of two intermediate levels of resistance. There was a significant increase in the frequency of isolates with higher levels of resistance to triadimenol during 1985. This is likely to have been a response to the continuing widespread use of demethylation inhibitor (DMI) fungicides by British farmers.     

Structure and evolution of a population of Erysiphe graminis f.sp. hordei

Samples of single colony isolates of Erysiphe graminis f.sp. hordei were collected in Cambridge on three dates in 1985, and tested for the presence of 12 virulence alleles and resistance to two fungicides, triadimenol and ethirimol. The frequency of the virulence V-(La) fell during 1985, while the frequency of V-h. virulence on cv. Triumph and higher levels of resistance to each fungicide and combined resistance to both fungicides rose. Two phenotypes, both of which possessed virulence on cv. Triumph and three unnecessary virulence alleles and had similar sensitivities to ethirimol, but differed in their level of resistance to triadimenol, accounted for 35.9% of all isolates. The high frequency of these phenotypes accounted for most of the observed gametic phase disequilibria between pathogenicity characters. Most individuals with these two phenotypes may be members of the same clone. It was estimated that 25% of the spore population which initiated the autumn epidemic of E.g. f.sp. hordei originated from ascospores formed by sexual reproduction in the summer. It is proposed that genetic drift followed by hitch-hiking selection, due to intense selection for a clone virulent on a newly-introduced cultivar, is a major factor influencing the frequency of unnecessary virulence alleles.     

白粉病菌侵染诱导的大麦叶肉细胞变化的超微结构与细胞化学

 从超微结构和细胞化学角度对大麦白粉病菌侵染诱导的寄主叶肉细胞变化进行了研究。结果发现,病原菌接种后12 h,叶肉细胞光合作用的标志性酶RCA和Rubisco含量发生迅速的变化,感病大麦(Ingrid)中RCA和Rubisco含量急剧减少,而抗病大麦(mlo-3)中则趋势相反。随后,不论在感病或抗病大麦中这2种蛋白的含量一直下降,但感病品种中一直维持相对较高水平。超微结构观察表明,病原菌侵染能够诱导侵染点附近叶肉细胞中叶绿体的增殖,新叶绿体的不断产生,从而延缓了叶肉细胞的衰老和死亡,增加了光合产物的形成。以上结果说明,叶肉细胞可能从养分供给上影响到了表皮细胞中病原菌吸器的发育。     

Utility of defeated resistance genes to powdery mildew,Erysiphe graminis f. sp. hordei,in spring barley variety mixtures

In 1980 and 1981 experiments have been performed to study the utility of defeated resistance genes to powdery mildew in spring barley variety mixtures. For this purpose the epidemic development of powdery mildew in pure stands and mixtures of four spring barley varieties was monitored. In three varieties the resistance was overcome several years ago. One variety is still resistant. Changes in frequencies of corresponding virulence genes were also studied.It was found that the variety mixtures slowed down the mildew epidemic only at the beginning of the season. Some reduction was lost later on since the pure stands reached their saturation level of infection at an earlier date than the more healthy mixtures. The level of reduction of infection depended on the composition of the mixtures and the number of components. Two-way mixtures appeared to be very risky due to the high frequency of corresponding combinations of virulence genes in the pathogen. Furthermore, two-way mixtures appeared to be unable to reduce the infection rate sufficiently when the infection pressure was high.In 1981 the frequencies of combinations of corresponding virulence genes increased very much in the Diva-Mazurka mixture but declined in the pure stands of both varieties.On the basis of these results it is concluded that the utility of defeated resistance genes involved in this study is very limited.The low number and the irregular distribution of the Dutch spring barley varieties over the different resistance groups prohibit the use of variety mixtures in the Netherlands.Samenvatting In 1980 en 1981 is onderzoek verricht over de bruikbaarheid van doorbroken resistentiegenen tegen meeldauw,Erysiphe graminis f. sp.hordei, in zomergerstrassenmengsels. Daartoe werd de epidemieontwikkeling in monocultures en mengsels van vier zomergerstrassen bestudeerd. Van drie rassen is de resistentie reeds verscheidene jaren doorbroken. Eén ras is nog resistent. De frequenties van compatibele meeldauwvirulentiegenen werden eveneens bepaald.Het bleek dat in vergelijking met de monocultures in de rassenmengsels de meeldauwepidemie alleen in het begin van het seizoen werd afgeremd. Later in het seizoen ging weer enige reductie in aantasting van de mengsels verloren doordat in de monocultures de aantasting eerder het verzadigingspunt bereikte dan in de minder aangetaste mengsels. De mate waarin de aantasting gereduceerd werd was afhankelijk van de aard en het aantal componenten in het mengsel. Twee-componentenmengsels bleken vrij riskant te zijn vanwege de aanwezigheid van combinaties van corresponderende virulentiegenen in de meeldauwpopulatie. Verder bleken tweecomponentenmengsels niet in staat de infectiesnelheid afdoende af te remmen wanneer de infectiedruk hoog werd.De frequentie van combinaties van corresponderende virulentiegenen nam sterk toe in het Diva-Mazurka-mengsel doch nam af in de monocultures van deze beide rassen.Gezien de verkregen resultaten lijkt de bruikbaarheid van de in dit onderzoek betrokken doorbroken resistentiegenen erg beperkt. Het geringe aantal zomergerstrassen op de rassenlijst en de ongelijke verdeling over de te onderscheiden resistentiegroepen belemmeren de toepassing van rassenmengsels in de Nederlandse akkerbouw.     

Comparison of resistance-inducing abilities of virulent and avirulent races of Erysiphe graminis f.sp. hordei and a race of Erysiphe graminis f.sp. tritici in barley

Resistance to powdery mildew was induced in barley by preinoculation with virulent and avirulent races of barley powdery mildew ( Erysiphe graminis f.sp. hordei ), and with a race of wheat powdery mildew ( E. graminis f.sp. tritici ). Four inducer densities were tested in 13 different induction periods between 1 and 24 h. Generally, the resistance induced by barley powdery mildew increased up to 10-12 h of induction and was maintained in longer induction periods. The inducing abilities of virulent and avirulent races could not be distinguished up to 10-12 h of induction, after which the inducing ability of avirulent races increased significantly in relation to virulent races. Wheat powdery mildew was able to induce more resistance than barley powdery mildew in induction periods up to 8 h. In a single inoculation procedure the number of haustoria developing from virulent barley powdery mildew decreased as inoculum density increased. The effect was ascribed to induction of resistance. This reduction of infection efficiency in the compatible interaction was compared to induced resistance. However, the inoculum density needed for 50% resistance induction in the double inoculation procedure was approximately 40 times higher than the inoculum density needed for 50% reduction in infection efficiency in the single inoculation procedure.     

Development of genetic SSR markers in Blumeria graminis f. sp. hordei and application to isolates from Australia

The barley powdery mildew pathogen, Blumeria graminis f. sp. hordei (Bgh), exists in numerous haplotypes and displays significant differences in fungicide sensitivity. It causes considerable yield losses throughout the world. Microsatellite SSRs are useful tools to study the population level and biogeographic aspects of intraspecific diversity, but so far none have been defined for Bgh. Here, eight polymorphic microsatellite loci were identified and characterized. Primer pairs amplifying the loci were then applied to 111 isolates of Bgh from Australia. The number of alleles per locus ranged from 4 to 13, and Nei's genetic diversity ranged from 0·25 to 0·76. The microsatellite primers detected several clones among the isolates and defined 97 unique haplotypes. There was little evidence for regional genotypic subdivision, suggesting that gene flow may not be restricted among geographic regions. All data was consistent with high levels of genetic diversity, potentially resulting from random mating and spread within each region.     

The effect of fungal density on fungicide dose–response curves in barley powdery mildew (Erysiphe graminis f.sp. hordei)

The effect of inoculation density on fungicide dose–response curves and the estimated ED₅₀ value (the fungicide concentration needed to halve the infected leaf area relative to the infected area of leaves that had not been treated with a fungicide) were investigated theoretically, and predictions were tested experimentally using powdery mildew of barley ( Erysiphe graminis [syn. Blumeria graminis ] f.sp. hordei ). A host leaf was assumed to consist of a number of independent compartments, in each of which only a single fungal spore could germinate successfully. The number of fungal spores landing in a compartment was assumed to be Poisson-distributed. The spores were assumed to vary in their sensitivity toward a fungicide, and the sensitivity of the spore population was assumed to be normally distributed around a mean sensitivity. Under these assumptions, the ED₅₀ value was shown to be a positively biased estimate of the mean sensitivity in the fungal population, and the bias increased with inoculum density. Consequently, the estimated ED₅₀ value is expected to vary between experiments and laboratories if the inoculation density varies over a considerable range. The correlation between inoculum density and estimated ED₅₀ value is expected to be strongest when the assayed fungal population consists of several genotypes differing in sensitivity. This expectation was tested by estimating the ED₅₀ values for one barley powdery mildew isolate at different inoculation densities for three different fungicides. These ED₅₀ values were positively correlated strongly with the inoculum density for fenpropimorph and more weakly for triadimenol, whereas no correlation was observed for propiconazole.     

Genetics of responses to morpholine-type fungicides and of avirulences inErysiphe graminis f. sp.hordei

The genetics of the responses of the barley powdery mildew pathogen,Erysiphe graminis f.sp.hordei, to three morpholine-type fungicides were studied. Resistances to a phenylpropylamine fungicide, fenpropidin, and to a morpholine, fenpropimorph, co-segregated in crosses of a sensitive isolate, DH14, with each of two resistant ones, CC151 and CC152. In the cross CC151×DH14, the results were consistent with resistance to both fungicides being controlled by a single gene, at a locus namedFenl. In the other cross, CC152×DH14, the genetics of resistance were more complicated; the data were consistent with the segregation of two complementary, unlinked genes which each conferred resistance to both fungicides. Fenpropidin-resistant progeny of CC151×DH14 were significantly more resistant to fenpropimorph than were fenpropidin-resistant progeny of CCI 52×DH14, although the resistant progeny of the two crosses did not differ significantly in their level of fenpropidin resistance. Fenpropidin-resistant progeny of CC151×DH14 were significantly more resistant to another morpholine, tridemorph, than were fenpropidin-sensitive progeny, but this was not the case for CC152×DH14. Resistance to triadimenol, a C14 demethylation-inhibitor (DMI) fungicide, segregated in both crosses. Triadimenol resistance appeared to be controlled by one gene in each cross and was not associated with morpholine resistance. CC151×DH14 also segregated for eight avirulence genes. Two of these matched theMla6 resistance, while one gene matched a previously unknown resistance in a Pallas near-isogenic line, P17, which also carries a known resistance gene,Mlk. Fenl was not significantly linked to the triadimenol resistance gene,Tdl(a), or to any of the eight avirulence genes.Avr _a6 1, Avr _a12 ,Avr _La ,Avr _p17 andTdl(a) were linked, as wereAvr _a 10 andAvr _k .Abbreviations ED₅₀ median effective dose - Fpd fenpropidin - Fpm fenpropimorph - PCA principal components analysis - Tdm tridemorph     

An apparatus for collecting total conidia of Blumeria graminis f.sp. hordei from leaf colonies using electrostatic attraction

Conidia from living conidiophores of barley powdery mildew ( Blumeria graminis f.sp. hordei ) on host leaves were collected consecutively using an electrostatic spore collector. The collector consisted of an electrical conductor plate linked to an electrostatic voltage generator and insulator plates placed abreast on a timed conveyer. The conductor plate was negatively charged by the potential supplied from the voltage generator. The negatively charged conductor plate caused dielectric polarization of the insulator plate, and the surface charge on the insulator plate attracted mature conidia abstricted from conidiophores on colonies growing on leaves placed 2 cm from the insulator plate. The surface charge on the insulator plate was proportional to the voltage applied to the conductor plate. Under optimized conditions, abstricted conidia were attracted to the electrostatically activated insulator plates without any detriment to their survival. During a colony's life span of c . 460 h, conidia were released throughout the day and c . 12 × 10⁴ conidia were collected during the lifetime of the colony. This is the first report on the direct quantification of progeny conidia produced by powdery mildew infecting host leaves.     

Racial diversity and complexity in regional populations of Erysiphe graminis f.sp. hordei in France over a 5-year period

The evolution of race patterns in three French regional populations of the barley powdery mildew pathogen Erysiphe graminis f.sp. hordei over a 5-year period showed rapid adaptation to newly introduced host resistance genes. In all three regions, the main change consisted of the replacement of initially abundant races by pathotypes differing markedly from them by their virulence gene combinations. This explained the increase in diversity during the first 3 years of the survey, when the second group of pathotypes became more common in the populations, and its subsequent decrease due to the decline of the first group of races. The mean number of virulence genes per isolate did not vary noticeably over time in the three populations, remaining at about four out of 12 genes tested. However, the distribution of the isolates into virulence complexity classes was greatly modified, fitting a binomial distribution by the end of the study, although significant deviations were apparent in the first 2 years (1986 and 1987). The data indicate that selection, migration and recombination are the most important factors shaping race structure and evolution in powdery mildew populations, and that mutation is of limited significance. No convincing evidence was obtained for the existence of stabilizing selection sensu Vanderplank as the mechanism limiting virulence complexity. Implications regarding spatial and temporal deployment of race-specific resistance genes to control powdery mildew are discussed.     

Effect of Film-forming Polymers on Infection of Barley with the Powdery Mildew Fungus, Blumeria graminis f. sp. hordei

The effects of three film-forming compounds, Ethokem, Bond and Vapor Gard, on infection of barley by the powdery mildew fungus Blumeria graminis f. sp. hordei were examined in glasshouse and field experiments. The three compounds provided significant control of powdery mildew infection when applied as pre- or post-inoculation treatments in the glasshouse. Such treatment had no effect on plant growth. Bond and Vapor Gard reduced the germination of conidia of B. graminis by 78% and 85% respectively, and reduced the subsequent formation of appressoria (73% and 85% respectively) and haustoria (75% and 79% respectively). The three compounds were less effective in field experiments, although they provided significant control of mildew infection and had no impact on plant growth and grain yield.     

Measuring the sterol content of barley leaves infected with powdery mildew as a means of assessing partial resistance to Erysiphe graminis f.sp. hordei

Barley leaf segments of several partially resistant cultivars were inoculated with different strains and combinations of strains of Erysiphe graminis. Infection level was assessed by counting colony number, by measuring sporulating area using an image analyser, and by extracting and quantifying sterols. Good correlations were obtained between data from all three methods. One cultivar contained less sterol than expected from the number of colonies present but some of the resistance in that cultivar was known to be due to reduced colony size. Image analysis was unable to discern the reduced colony size component of resistance. Scanning electron microscopy revealed that single colonies may originate from more than one conidial infection. Sterol analysis was the most representative method of comparing the reaction of diverse host genotypes to mildew infection.     

Virulence analysis of Puccinia graminis f.sp. tritici populations in Ethiopia with special consideration of Ug99

Wheat stem rust samples were collected in 2006 and 2007 in the Arsi, Bale, Shewa and northwest regions of Ethiopia to determine virulence diversity and race distribution in Puccinia graminis f.sp. tritici populations. Stem rust incidence was high in Arsi, Bale and east Shewa. In northwest Ethiopia, and north and west Shewa, stem rust was prevalent at low levels. A total of 152 isolates was analysed and 22 races were identified. Races TTKSR (Ug99), TTHSR and RRTTR were predominant, with frequencies of 26·6, 17·7 and 11·1%, respectively. These races were also detected in all regions. The highly virulent race designated Ug99 was present throughout the country and dominated in all regions except northwest Ethiopia. A variant of Ug99 virulent against the stem rust resistance gene Sr24 was not detected in this study. Four stem rust resistance genes ( Sr13, Sr30, Sr36 and SrTm p) were found to confer resistance to most of the races prevalent in Ethiopia. With the exception of Sr30 , which is not effective against Ug99, these genes could be used in breeding for resistance to stem rust in Ethiopia.     

Baseline sensitivity to proquinazid in Blumeria graminis f. sp. tritici and Erysiphe necator and cross‐resistance with other fungicides

BACKGROUND: Proquinazid is a new quinazolinone fungicide from DuPont registered in most European countries for powdery mildew control in cereals and vines. The aim of this paper is to present baseline sensitivity data in populations of Blumeria graminis f. sp. tritici EM Marchal and Erysiphe necator (Schw) Burr as well as results from cross‐resistance studies with other fungicides. RESULTS: Proquinazid exhibited a high intrinsic activity on B. graminis f. sp. tritici isolates at rates ranging from 0.000078 to 0.02 mg L^?1. Erysiphe necator isolates were comparatively less sensitive to proquinazid, with EC₅₀ values ranging from 0.001 to 0.3 mg L^?1. Proquinazid controlled equally well B. graminis f. sp. tritici isolates sensitive and resistant or less sensitive to tebuconazole, fenpropimorph, fenpropidin, cyprodinil and kresoxim‐methyl. A positive correlation (r = 0.617) between quinoxyfen and proquinazid sensitivities was found among 51 B. graminis f. sp. tritici isolates. Quinoxyfen‐resistant B. graminis f. sp. tritici isolates were slightly less sensitive to proquinazid than the quinoxyfen‐sensitive isolates; however, proquinazid remained much more active than quinoxyfen on these isolates. A stronger sensitivity relationship (r = 0.874) between proquinazid and quinoxyfen was found among 65 E. necator isolates tested in a leaf disc assay. The sensitivity values for proquinazid were significantly lower than those for quinoxyfen, confirming the higher intrinsic activity of proquinazid on both pathogens. CONCLUSION: Given the history of resistance development in powdery mildew and the observed sensitivity relationship with quinoxyfen, specifically in E. necator, we conclude that the risk of resistance developing to proquinazid might be influenced by the use of quinoxyfen. Based on these results, the authors recommend that proquinazid and quinoxyfen be managed together for resistance management. Copyright © 2009 Society of Chemical Industry     

甘肃省小麦白粉菌群体毒性及小麦品种苗期抗白粉病性分析

由Blumeria graminis f.sp.tritici引起的小麦白粉病是危害甘肃省小麦安全生产的重要病害之一,分析病原菌毒性结构和抗病基因有效性对于指导白粉病的有效控制具有重要意义。2013年对甘肃省小麦白粉菌群体进行毒性分析结果表明,91个供试菌系中,对Pm1、Pm2、Pm3a、Pm3b、Pm3c、Pm3d、Pm3e、Pm3f、Pm4a、Pm4b、Pm5a、Pm6、Pm7、Pm8、Pm19、Pm33、Pm5+Pm6、Pm4+Pm8、Pm4b+Pm5b和PmEra的毒力频率达到70%以上,已无利用价值;对Pm13、Pm16、Pm21和Pm24的毒性频率在15%以下,尚可利用。选用甘肃省不同毒性谱的单孢堆菌系,对35个甘肃省及国内生产品种(系)进行苗期致病性测定,发现仅有‘绵麦37’具有优异抗病性。供试白粉菌系对‘定西40号’等22个品种(系)的毒性频率达到60%以上,甘肃省及中国生产品种中,苗期抗病品种(系)匮乏。     

The isolation of a further new race of Erysiphe graminis DC. f.sp. hordei Marchal and the genetical basis of the resistance of ‘Lyallpur 3645’

Besides the race Amsel-C₂, a new race of barley mildew (Erysiphe graminis DC. f.sp.hordei Marchal) has been isolated from a still resistant variety L 94. This race has provisionally been denoted Lyallpur 3645-C₁₇. It is able to overcome the resistance of Lyallpur 3645.It appears most likely that the barley variety Lyallpur 3645 has two closely linked genes for resistance, one for hypersensitivity and the other for an intermediate type of reaction. The first is present in the variety Amsel and both in Heine 4808.The reactions indicate that race Lyallpur 3645-C₁₇ has only one gene for virulence in addition to the ones present in race Amsel-C₂.A survey is given of the seedling reaction of some 60 previously very or moderately resistant varieties, after infection with the two new races.Samenvatting Twee fysio's van gerstemeeldauw,Erysiphe graminis DC. f.sp.hordei Marchal zijn gevonden, waarvan één niet eerder is beschreven. Dit laatste fysio wordt voorlopig aangeduid als Lyallpur 3645-C₁₇, het andere is het zgn. Amsel-C₂-fysio.Met behulp van beide fysio's is duidelijk geworden dat de lijn Lyallpur-3645 twee nauw gekoppelde resistentiegenen bezit; een gen voor een overgevoeligheidsreactie en een ander gen voor een intermediaire reactie.In het ras Amsel is alleen het overgevoeligheidsgen aanwezig, in Heine 4808, evenals Amsel afkomstig van kruisingen met Lyallpur 3645, zijn beide genen aanwezig.Het fysio Lyallpur-3645-C₁₇ lijkt daarom één gen voor virulentie meer te hebben dan Amsel-C₂.Tenslotte is een overzicht gegeven van de kiemplantreactie na infectie met beide fysio's van ongeveer 60 gerstrassen die eerder zeer goed of matig resistent waren.