Variation in sensitivity to fungicides which inhibit ergosterol biosynthesis in wheat powdery mildew

Fungicides which inhibit ergosterol biosynthesis have been in use for control of wheat powdery mildew (Erysiphe graminis f. sp.tritici) in the Netherlands since 1978. Mildew populations were tested for their variation in sensitivity to triadimefon from 1982 to 1984.In 1982 isolates from the province Limburg, with a triazole spray-regime history, were less sensitive to triadimefon than isolates from the provinces Gelderland and Noord-Brabant, where triazoles had not been used. In the following years isolates with reduced sensitivity were also detected in the latter provinces and other parts of the country. This spread correlates with the increased use of triazoles, both in frequency and space, from 1983 onwards. The reduced sensitivity can as a whole or in part be responsible for the decline in field performance of triazoles, observed during these years.Cross-sensitivity to the triazoles triadimefon and propiconazole was established, but not to triazoles and the morpholine fungicide fenpropimorph. Effectiveness of the latter compound was similar to all isolates from Limburg tested in 1984. Field performance of fenpropimorph, introduced in 1983, appeared to be normal. It is recommended to counteract further development of resistance by sequential use of fenpropimorph early in the season (May) and triazoles at the end (June–July).Samenvatting Fungiciden die de ergosterolbiosynthese remmen worden in Nederland sinds 1978 gebruikt bij de bestrijding van tarwemeeldauw (Erysiphe graminis f. sp.tritici) Meeldauwpopulaties werden getoetst op hun variatie in gevoeligheid voor triadimefon van 1982 tot 1984.In 1982 bleken isolaten afkomstig uit de provincie Limburg, waar voordien triazolen werden toegepast, minder gevoelig te zijn voor triadimefon dan isolaten uit de provincies Gelderland en Noord-Brabant, waar nog geen triazolen werden gebruikt. In de daaropvolgende jaren werden isolaten met een verminderde gevoeligheid ook in laatstgenoemde provincies en in andere delen van het land gevonden. Deze uitbreiding is gecorreleerd met een toename in het gebruik van triazolen vanaf 1983 in areaal en frequentie. De afname in gevoeligheid kan geheel of gedeeltelijk verantwoordelijk zijn voor de verminderde meeldauwwerking van triazolen die gedurende deze jaren werd waargenomen.Kruisgevoeligheid werd vastgesteld voor de triazolen triadimefon en propiconazool, maar niet voor triazolen en het morfoline-derivaat fenpropimorf. De werking van dit middel was tegen alle isolaten uit Limburg die in 1984 werden getoetst, gelijk. De meeldauwwerking van fenpropimorf, dat in 1983 werd geïntroduceerd, was normaal. Aanbevolen wordt om verdere resistentie-ontwikkeling tegen te gaan door afwisselend gebruik van fenpropimorf vroeg in het groeiseizoen (mei) en een triazool aan het eind (juni-juli).     

防治西葫芦和黄瓜白粉病的生物制剂的筛选

瓜类白粉病是蔬菜上的重要病害之一,为加大对该病害的生物防治力度,本研究采用课题组研发的1×109cfu/g玫瑰黄链霉菌水剂和10亿芽胞/g枯草芽胞杆菌悬浮剂以及市场上常用的枯草芽胞杆菌、哈茨木霉菌、寡雄腐霉、武夷菌素、多抗霉素等多种生物制剂进行田间药效试验,比较几种生物制剂的防治效果。结果表明,供试生物制剂对西葫芦白粉病的防效为57.65%~84.98%,其中3%多抗霉素水剂600倍液的防效最好,为84.98%,且具有明显的促生长作用,增产率达12.65%。其次为1 000亿芽胞/g枯草芽胞杆菌可湿性粉剂(武汉天惠)400倍液、2%武夷菌素水剂600倍液和3×108 cfu/g哈茨木霉菌可湿性粉剂300倍液,可作为西葫芦白粉病防治的选用药剂。1×109 cfu/g玫瑰黄链霉菌水剂对西葫芦和黄瓜白粉病均具有较好的防效,分别为78.68%和73.59%,具有开发和应用的市场价值。     

Mining wild barley for powdery mildew resistance

Powdery mildew, caused by Blumeria graminis f. sp. hordei (Bgh), is a worldwide disease problem on barley (Hordeum vulgare) with potentially severe impact on yield. Historically, resistance genes have been identified chiefly from cultivated lines and landraces; however, wild barley (H. vulgare subsp. spontaneum) accessions have proven to be extraordinarily rich sources of powdery mildew resistance. This study describes the characterization of a collection of 316 wild barley accessions, known as the Wild Barley Diversity Collection (WBDC), for resistance to powdery mildew and the genetic location of powdery mildew resistance loci. The WBDC was phenotyped for reaction to 40 different Bgh isolates at the seedling stage and genotyped with 10 508 molecular markers. Accessions resistant to all 40 isolates of Bgh were not found; however, three accessions (WBDC 053, 085 and 089) exhibited resistance to 38 of the isolates. Gene postulation analyses revealed that many accessions, while resistant, contained none of the 12 genes present in the Pallas near‐isogenic lines Mla1, Mla3, Mla6, Mla7, Mla9, Mla12, Mla13, Mlk1, MlLa, Mlg, Mlat and Ml(Ru2), suggesting that the accessions carry novel genes or gene combinations. A genome‐wide association study of powdery mildew resistance in the WBDC identified 21 significant marker‐trait associations that resolved into 15 quantitative trait loci. Seven of these loci have not been previously associated with powdery mildew resistance. Taken together, these results demonstrate that the WBDC is a rich source of powdery mildew resistance, and provide genetic tools for incorporating the resistance into barley breeding programmes.     

Spatial aggregation of pathotypes of barley powdery mildew

Aggregation in the distribution of pathotypes of Erysiphe graminis f.sp. hordei , the barley powdery mildew pathogen, was investigated in field plots of 'Golden Promise', 'Proctor' and 'Tyra'. 'Golden Promise' and 'Proctor' have no effective mildew resistance alleles, whereas 'Tyra' has Mla1 , which was only effective against a proportion of the mildew population. Isolates of mildew were sampled according to a grid sampling scheme and their virulence spectra ascertained in order to group them according to pathotype. The populations were very diverse, and evidence for aggregation (quantified using join counts) was found only in the 'Tyra' plots, at distances of up to 1m. This aggregation was reduced in a subsequent sample. The results are consistent with a model in which mildew epidemics are started by a large number of initial infections, which then form diffuse, overlapping aggregations of clones. These aggregations then become more diffuse, so that the amount of aggregation reduces with time. The greater amount of aggregation seen in the 'Tyra' plots might have been caused by there being less initial inoculum with virulence towards that cultivar.     

Movement of barley powdery mildew within field plots

Movement of barley powdery mildew (caused by Erysiphe graminis f.sp. hordei ) within fields was investigated by sowing the barley cultivars Tyra and Jupiter side by side in two field plots, and trapping spores along transects within the plots. The trapped spores were tested for virulence on the two cultivars. The epidemic on Tyra developed quickly, and a gradient in the proportion of spores with virulence on Tyra was detected in the Jupiter half-plots. In the Jupiter half-plots, the epidemic was much less severe; and no mildew could be found in one plot. Movement of spores from one half of the plot to the other usually declined steeply in the first 4 m from the boundary, and was not detectable beyond 12 m. There were exceptions where the gradient was much shallower, and these were consistent with differences in wind direction.     

Interaction between powdery mildew and barley with mlo5 mildew resistance

Powdery mildew infection of barley with the mlo5 barley powdery mildew resistance gene was examined, using near-isogenic barley lines, with and without mlo5 resistance, and two near-isogenic powdery mildew isolates, HL3/5 and GE3 with high (virulent) or low (avirulent) penetration efficiency on the resistant barley line. In all isolate–host combinations (except GE3 on the resistant barley line), frequency of haustorium formation increased significantly from zero at 11 h after inoculation to a maximum by 13 h, and there was no subsequent increase up to 24 h. In the susceptible barley line, 27% of appressoria from both isolates formed haustoria. Although this was significantly higher than the frequency of haustorium formation (18%) of HL3/5 on the resistant barley line, HL3/5 was much more successful than GE3 (frequency of haustorium formation less than 1%). The fact that HL3/5 did not possess a generally higher ability to penetrate successfully to form haustoria on the susceptible barley line, indicates that HL3/5 did not overcome the mlo5 resistance by being generally more vigorous. In the resistant barley line, papillae were larger than in the susceptible line; however, both isolates were associated with papillae of the same diameter at the time of penetration. We suggest that the mlo5 resistant barley line confers two different forms of resistance: isolate-specific and isolate-nonspecific.     

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.     

Potassium phosphate induces systemic protection in barley to powdery mildew infection

In laboratory tests, treatment of the first leaves of barley (Hordeum vulgare L cv Golden Promise) with potassium phosphate led to significant reduction in infection of the second leaves with the powdery mildew fungus Blumeria graminis f sp hordei Marchal, with a 25 mM treatment giving 89% reduction in infection. Although the optimal interval between phosphate treatment of the first leaves and mildew inoculation of the second leaves was 2 days, significant protection was still obtained if the interval was increased to 12 days. Protection against powdery mildew infection was not as effective when the potassium phosphate was applied as a seed treatment or root drench. Phosphate treatment of the first leaves led to significant increases in activities of phenylalanine ammonia lyase (PAL), peroxidase and lipoxygenase in second leaves. Enzyme activities, especially PAL and peroxidase, were increased further when second leaves of phosphate-treated plants were inoculated with powdery mildew. Phosphate treatment of the first leaves did not adversely affect plant growth and, in a field trial, 25 mM potassium phosphate provided 70% control of mildew and gave a small increase in grain yield.     

Effects of barley genotype on induced resistance to powdery mildew

Infection of barley by an avirulent isolate of Erysiphe graminis f.sp. hordei reduces the development of powdery mildew disease by a virulent isolate, inoculated subsequently. This phenomenon, known as induced resistance, was studied in three sets of near-isogenic lines of barley, each of which included varieties with four different race-specific resistance genes. The extent of induced resistance, expressed as a reduction in the number of colonies formed, differed in lines carrying different race-specific resistance genes, being most effective in lines with Mla7 , followed by Mth , then Mla6 and Mla13. These differences may be related to the extent of the hypersensitive response to infection by avirulent spores. A further effect of induced resistance in reducing sporulation, taking the number of colonies into account, was least effective in Mth lines, with the other lines ranking as they did when induced resistance was expressed as colony numbers. By contrast, the genetic backgrounds of the three near-isogenic sets had similar effects on the fractional reduction of colony number by induced resistance. The effect of induced resistance on sporulation was strongest in the most resistant background, and weakest in the most susceptible.     

Control of cucumber powdery mildew by the hyperparasite Ampelomyces quisqualis and fungicides

Application of conidial suspensions of the hyperparasite Ampelomyces quisqualis to greenhouse cucumbers inoculated with Sphaerotheca fuliginea increased cucumber yield compared with an untreated control. Similar yield increases were obtained with triforine and also when a reduced rate of triforine and the hyperparasite were combined. A. quisqualis extensively parasitized S. fuliginea and also, in experiments with commercial cucumber crops, Erysiphe cichoracearum.     

Factors affecting build-up of fungicide resistance in powdery mildew in spring barley

In replicate-plot field experiments done in the UK, at one site in Avon for 3 years and another in Warwickshire for 2 years, application of ethirimol or triadimenol sprays or seed treatments was followed by decreases in sensitivity of mildew samples to the particular fungicide applied. Application of ethirimol-triadimenol or tridemorph-triadimenol mixtures caused smaller or no decreases in sensitivity. Differences between isolates in responses to ethirimol and to triadimenol treatment were usually negatively correlated. Early-season inoculum differed in fungicide sensitivity between sites. At one site sensitivity shifted markedly from one season to another. No clear interactions between cultivar, mildew pathotype and shifts in fungicide response could be discerned. There were no major differences in resistance build-up between seed or spray treatments.     

Factors affecting build-up of fungicide resistance in powdery mildew in spring barley

In replicate-plot field experiments done in the UK, at one site in Avon for 3 years and another in Warwickshire for 2 years, application of ethirimol or triadimenol sprays or seed treatments was followed by decreases in sensitivity of mildew samples to the particular fungicide applied. Application of ethirimol-triadimenol or tridemorph-triadimenol mixtures caused smaller or no decreases in sensitivity. Differences between isolates in responses to ethirimol and to triadimenol treatment were usually negatively correlated. Early-season inoculum differed in fungicide sensitivity between sites. At one site sensitivity shifted markedly from one season to another. No clear interactions between cultivar, mildew pathotype and shifts in fungicide response could be discerned. There were no major differences in resistance build-up between seed or spray treatments.In veldexperimenten met herhalingen uitgevoerd in Engeland, gedurende een periode van 3 jaar in Avon en gedurende een periode van 2 jaar in Warwickshire, werd na behandeling van zomergerst met ethirimol of triadimenol een verminderde gevoeligheid van meeldauw voor deze fungiciden waargenomen.Behandelingen met mengsels van ethirimol-triadimenol of tridemorf-triadimenol gaven weinig tot geen verminderde gevoeligheid. Verminderde gevoeligheid van isolaten voor ethirimol was meestal gecorreleerd met een verhoogde gevoeligheid voor triadimenol en omgekeerd. Vroeg in het seizoen werd in de meeldauwpopulatie op de twee proefvelden een verschil in gevoeligheid voor de fungiciden waargenomen. Op één proefveld trad van het ene op het andere seizoen een aanzienlijke verandering in de gevoeligheid voor de fungiciden op. Er was geen duidelijke correlatie tussen de waargenomen verminderde gevoeligheid voor de fungiciden en de gebruikte cultivars of voorkomende fysio's. Verminderde gevoeligheid voor de fungiciden werd zowel bij zaadbehandeling als bij het bespuiten van planten waargenomen.     

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.     

Local and systemic effects of oxylipins on powdery mildew infection in barley

Treatment of the first leaves of barley seedlings with the oxylipin colneleic acid, or the two trihydroxy oxylipins 9,12,13-trihydroxy-11(E)-octadecenoic acid and 9,12,13-trihydroxy-10(E)-octadecenoic acid, reduced infection of that leaf by the powdery mildew fungus Blumeria graminis Speer f sp hordei Marchal. When applied to first leaves, etheroleic acid and colneleic acid, as well as the trihydroxy oxylipin 9,12,13-trihydroxy-10(E),15(Z)-octadecadienoic acid, also reduced mildew infection in second leaves. In all cases where local and systemic effects against mildew were observed, activity of the defence-related enzyme phenylalanine ammonia lyase (PAL) was increased, but only following challenge inoculation with powdery mildew. Peroxidase activity was not affected by oxylipin treatment or mildew inoculation. Whether the effects observed were due to the oxylipins or to breakdown products is not known, since no information is available on the stability of these particular oxylipins on leaf surfaces. Nevertheless, these data represent the first report of systemic effects against pathogen infection following pre-treatment with oxylipins.     

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.     

Effects of recombination on races of a barley powdery mildew population

Analyses of virulence were performed with isolates from ascospore populations and their possible parental populations in order to measure the potential effect of recombination on the pathogenic variation in populations of Erysiphe graminis f. sp. hordei. The conidial isolates and cleistothecia were sampled from a field of spring barley cv. Golden Promise. Compared to the summer conidial populations, higher frequencies of rare races and rare virulence factors together with greater diversity were observed among progeny from ascospores, and the most frequent race was less abundant. The contribution of ascospores to primary infection of barley in autumn could not be measured directly, but analyses of the conidial populations on volunteer seedlings of Golden Promise suggested that sexual recombination was one of the mechanisms that had caused the change in population structure.     

Effects of fungicides on powdery mildew, tree growth and cropping of apple

In a 6-year orchard experiment, seven fungicide programmes were assessed for control of powdery mildew (Podosphaera leucotricha) and for long-term effects on growth and cropping of the apple cultivar Cox's Orange Pippin. Programmes based on binapacryl, bupirimate, fenarimol, nitrothal-isopropyl and triadimefon gave better control than dinocap or triforine. The heaviest cumulative yields of marketable fruit were obtained with the non-systemic fungicides binapacryl and nitrothal-isopropyl, the former significantly outyielding triadimefon and triforine over 6 years. Tree vigour, as assessed by extension shoot length and internode length, was found to be better after 4-5 years of programmes based on binapacryl than on triadimefon or fenarimol. A relation was demonstrated between cumulative marketable yield and mean annual mid-season incidence of secondary mildew on extension shoot leaves.     

Resistance to powdery mildew in one Spanish barley landrace hardly resembles other previously identified wild barley resistances

Two major quantitative trait loci (QTLs) associated with resistance to powdery mildew (Blumeria graminis f. sp. hordei) were previously identified on chromosome 7H of the Spanish barley line SBCC097. The two QTLs seemed to share the same chromosomal position as the major genes mlt and Mlf, which were formerly described in Hordeum vulgare ssp. spontaneum-derived lines. In the present work, different lines that carry mlt (RS42-6*O), Mlf (RS137-28*E), or a combination of both (SI-4 and SI-6) were compared with SBCC097 to evaluate their relatedness at the phenotypic, cellular, and genetic levels. The resistance of the lines was characterised by inoculating them with a set of 27 isolates of B. graminis, which displayed a wide range of virulence. It was revealed that SBCC097 possessed a distinctive resistance spectrum. Microscopic assessment of the cytological development of the resistance response showed that SBCC097 clearly formed fewer well-established colonies and secondary hyphae than the other lines. This was confirmed by the infection type recorded after visual inspection. Genetic analyses of all five lines, based on markers flanking the QTLs derived from SBCC097, supported the macroscopic and microscopic data and pointed to the presence of a combination of novel genes or alleles in SBCC097, which may be included in the category of “intermediate-acting” genes, governing resistance mainly at the post-penetration stage.     

Tomato powdery mildew

A powdery mildew ( Erysiphe sp.) occurred on greenhouse-grown tomato crops in southern England in 1987. White pustules appeared on the upper surfaces of leaves of affected crops and only rarely on the under surfaces. In greenhouse experiments, stems were very severely affected. In host range studies 19 tomato cultivars and six breeders' lines were susceptible as well as Nicotiana tabacum. N. xanthi. Solanium melongena, S. pseudocapsicum, S. tuberosum, Datura stramonium and Petunia hybrida. Conidia germinated readily at 15, 20 and 25 C and were viable for 2 days in the laboratory and at least 6 days in the greenhouse. The disease was well controlled by a range of fungicides including benomyl, bupirimate, carbendazim, chlorothalonil, fenarimol and pyrazophos. All of these except pyrazophos are approved for use on tomatoes in the UK. The pathogen was very sensitive to low concentrations of benomyl and fenarimol but less so to bupirimate.