Ethirimol-sensitivity in populations of Erysiphe graminis from plots of spring barley

Barley plots, cv. Proctor were sown with either ethirimol-treated or untreated seed. At seedling emergence, plots were inoculated with isolates of Erysiphe graminis either sensitive or insensitive to ethirimol, or left uninoculated. Mildew samples taken from the coleoptile leaves of seedlings, and later from leaf 6 (next but one to the flag leaf) were compared for ethirimol sensitivity using probit analysis. Probit lines were derived using ‘Wadley's analysis’. This analysis uses the number of survivors to estimate the total number of individuals originally present. In the test for ethirimol sensitivity a count of colonies on inoculated test leaves was taken to represent survivors. Microscopic counting of the original inoculum was thus avoided. The analyses snowed that the initial leaf inoculation treatments established mildew populations in the plots, which differed considerably in ethirimol sensitivity. Some 6 wk after inoculation the levels of sensitivity in these populations remained unchanged even though inoculum coming into the experiment from external sources was often much more insensitive than the mildew in some plots. Probit lines of response to ethirimol had shallow slopes, showing that large variability exists in the pathogen for this character.     

Studies on the tolerance of Erysiphe graminis f. sp. hordei to systemic fungicides

Successive sowings of glasshouse-grown barley plants were treated with either tridemorph, ethirimol or both fungicides, and inoculated with an initially fungicide-sensitive isolate of Erysiphe graminis f. sp. hordei. The time taken for symptoms to appear, compared with that for untreated plants, decreased with successive sowings. This was interpreted as evidence for the increase in the frequency of fungicide-tolerant propagules in the pathogen population. Effective mildew control was obtained by the use of either or both fungicides in trial plot and field crops. Seedling assays for tolerance to tridemorph and ethirimol showed that tolerance was more evident in treated than in untreated crops in June. Some mildew populations partially tolerant to one fungicide also showed reduced sensitivity to the other. By July a response intermediate between tolerant and sensitive was recorded on all the plots, probably indicating the mixing of fungal populations from adjacent plots. Larger numbers of tolerant isolates were found in random samples from treated than from untreated crops.     

Progress of infection by Erysiphe graminis on fungicide-treated and untreated barley

Tridemorph-treated and untreated barley plants wre grown either in separate growth rooms or together. They were inoculated either with a sensitive or a tolerant isolate, and the progress of mildew was recorded. Comparisons were made between the rates of progress of sensitive and tolerant isolates on separated and unseparated fungicide treatments. Results suggested that interactions took place between pathogen populations on adjacent treated and untreated plants. Barley cv. Golden Promise was sown in plots made up of two adjacent fungicide treatments. Triadimefon, triforine, tridemorph and ethirimol were paired with each other and with untreated crops in all combinations. The progress of mildew on each plot was recorded, and differences were found between similar fungicide treatments from different plot pairs. The likelihood of interactions between mildew populations exposed in the field to different fungicide treatments is discussed.     

IV. Variation in fungicide tolerance: Variation in Erysiphe graminis f. sp. hordei in response to the fungicide ethirimol

The response of a fungal population to a fungicide is normally highly variable. Individual lethal doses usually form a normal distribution when plotted against log dose. The shape of this distribution varies with chemical and organism. Spore germination, haustorial formation, etc. vary independently. Variation in disease control is the product of the variation at each stage. Variation is both genotypic and phenotypic. The successful use of a fungicide should, theoretically, result in a shift in the dose-response of the survivors. The shift ought to be greatest where the original spread of the lethal dose was widest. The effect of ethirimol on barley powdery mildew supports these theoretical conclusions. Good control of mildew following the use of ethirimol resulted in residual populations which were less sensitive to the fungicide than those in nearby untreated fields. Within treated fields the most sensitive members of the population were absent and the overall variation decreased. The progressive shift in sensitivity which might have been expected did not occur. Successive sampling of the same fields revealed rapid changes in sensitivity, but in either direction. The proportion of less sensitive individuals declined toward the end of the season and much of the original variability returned.     

Regression-Based Ranking of Pathogen Strains with Respect to Their Contribution to Natural Epidemics

Genetic variation in pathogen populations may be an important factor driving heterogeneity in disease dynamics within their host populations. However, to date, we understand poorly how genetic diversity in diseases impact on epidemiological dynamics because data and tools required to answer this questions are lacking. Here, we combine pathogen genetic data with epidemiological monitoring of disease progression, and introduce a statistical exploratory method to investigate differences among pathogen strains in their performance in the field. The method exploits epidemiological data providing a measure of disease progress in time and space, and genetic data indicating the relative spatial patterns of the sampled pathogen strains. Applying this method allows to assign ranks to the pathogen strains with respect to their contributions to natural epidemics and to assess the significance of the ranking. This method was first tested on simulated data, including data obtained from an original, stochastic, multi-strain epidemic model. It was then applied to epidemiological and genetic data collected during one natural epidemic of powdery mildew occurring in its wild host population. Based on the simulation study, we conclude that the method can achieve its aim of ranking pathogen strains if the sampling effort is sufficient. For powdery mildew data, the method indicated that one of the sampled strains tends to have a higher fitness than the four other sampled strains, highlighting the importance of strain diversity for disease dynamics. Our approach allowing the comparison of pathogen strains in natural epidemic is complementary to the classical practice of using experimental infections in controlled conditions to estimate fitness of different pathogen strains. Our statistical tool, implemented in the R package StrainRanking, is mainly based on regression and does not rely on mechanistic assumptions on the pathogen dynamics. Thus, the method can be applied to a wide range of pathogens.     

Effects of chemical treatments for mildew control at different times on the growth and yield of spring barley

Experiments in 1973 and 1974 tested the effects of ethirimol seed treatment and the timing of single sprays of ethirimol or tridemorph, on mildew control and the growth and yield of spring barley. These treatments were compared with selected combinations of two treatments designed to give longer protection. In 1973, combinations of seed treatment and spray, or two sprays, did not yield better than the best component treatment applied alone or the most effective single spray of tridemorph applied on 1 June. In 1974, mildew developed usually early and yields were increased by applying ethirimol to the seed plus a tridemorph spray or two tridemorph sprays. However, barley sprayed once with tridemorph on 20 May, the expected best date, yielded more than barley receiving a seed treatment of ethirimol plus a tridemorph spray applied on either 3 or 12 June. Sprays of captafol and tridemorph, applied as separate treatments, successively to the same plots on three occasions, gave the best yields in both years. Treatments most affected ear number but they did have some effect on all components of yield. In 1974 there was a significant relationship between ear number and the variance of the number of grains per ear.     

Are plant pathogen populations adapted for encounter with their host? A case study of phenological synchrony between oak and an obligate fungal parasite along an altitudinal gradient

Biotrophic fungal pathogens are expected to have adapted to their host plants for phenological synchrony, to optimize the possibility of contacts leading to infections. We investigated the patterns and causes of variation in phenological synchrony in the oak‐powdery mildew pathosystem, a major disease in natural ecosystems. The study was carried out along an altitudinal gradient, representing a wide temperature range, in mature oak stands. Both sporulation (pathogen infective stage) and oak flushing (host susceptible stage) were delayed with increasing elevation, but with a significantly different sensitivity for the two species. This resulted in a variable host–pathogen synchrony along the gradient. A common garden experiment did not give evidence of among‐population genetic differentiation (past adaptation) for fungal phenology. This could be explained by the high phenotypic variation in phenology within host populations, precluding selection on fungal phenology at the population scale, but possibly favouring adaptation at the within‐population scale. Phenotypic plasticity was the major cause of the observed variation in the phenology of the fungal populations.     

Effects of mildew on the growth and yield of spring barley: 1969-72

Mildew decreased grain yield by decreasing ear number and grain size (but not grain number/ear), the damage depending on the earliness and severity of mildew. In 1971 when mildew was early and severe, ethirimol seed dressings at 0.22 kg a.i./ha, which gave only early protection, increased yield more than did ethirimol sprays applied to protect the flag leaf and ear. In 1972 sprays were better than seed dressings at this rate because mildew was less severe during the seedling stages.     

Intraepidemic heterogeneity of influenza A (H3N2) viruses in 1985: antigenic analysis and sensitivity to non-specific inhibitors

During the influenza outbreak of 1984-85 22 strains of H3N2 viruses were isolated in Finland. An intra-epidemic heterogeneity was demonstrated in an antigenic analysis by haemagglutination inhibition test with antisera produced in rats. The strains could be classified into three groups which corresponded to the following reference strains: group I: A/Hong Kong/1/84, A/Hong Kong/3/84; group II: A/Philippines/2/82; group III: A/Caen/1/84. Seven of the isolates were entirely insensitive to gamma-inhibitors of guinea-pig sera, which is in contrast to the small number of these viruses found among H3N2 strains isolated in the 1970s. The insensitive strains could not be isolated until the second or third passage through the eggs, whereas about half of the sensitive and intermediate strains were already isolated during the first passage. Conversions in reactivity with gamma-inhibitors could be detected only from an intermediate or an insensitive virus to a sensitive virus when several strains were passed serially in ovo and in MDCK cultures. The findings suggest that the gamma-inhibitor-insensitive strains corresponded well to the viruses of the human host or arose from dimorphic virus populations under an arbitrary selection of terminal dilution conditions prevailing during isolation in eggs. The insensitive strains did not differ substantially from the sensitive viruses in their ability to agglutinate erythrocytes of different laboratory animals or in their disagglutination patterns. On the other hand, propagation of viruses in MDCK cultures had an effect on these properties. The results are discussed with respect to Q phase variants and receptor binding properties.     

High intraspecific recombination rate in a native population of Candidatus pelagibacter ubique (SAR11)

Recombination is an important process in microbial evolution. Rates of recombination with extracellular DNA matter because models of microbial population structure are profoundly influenced by the degree to which recombination is occurring within the population. Low rates of recombination may be sufficient to ensure the lateral propagation of genes that have a high selective advantage without disrupting the clonal pattern of inheritance for other genes. High rates of recombination potentially can obscure clonal patterns, leading to linkage equilibrium, and give microbial populations a population genetic structure more akin to sexually interbreeding eukaryotic populations. We examined eight loci from nine strains of candidatus Pelagibacter ubique (SAR11), isolated from a single 2L niskin sample of natural seawater, for evidence of genetic recombination between strains. The Shimodaira-Hasegawa test revealed significant phylogenetic incongruence in seven of the genes, indicating that frequent recombination obscures phylogenetic signals from the linear inheritance of genes in this population. Statistical evidence for intragenic recombination was found for six loci. An informative sites matrix showed extensive evidence for a widespread breakdown of linkage disequilibrium. Although the mechanisms of genetic transfer in native SAR11 populations are unknown, we measured recombination rates, rho, that are much higher than point mutation rates, theta, as a source of genetic diversity in this clade. The eukaryotic model of species sharing a common pool of alleles is more apt for this SAR11 population than a strictly clonal model of inheritance in which allelic diversity is controlled by periodic selection.     

Origins and colonization history of pandemic Vibrio parahaemolyticus in South America

The dynamics of dissemination of the environmental human pathogen Vibrio parahaemolyticus are uncertain. The O3:K6 clone was restricted to Asia until its detection along the Peruvian coasts and in northern Chile in 1997 in phase with the arrival of El Niño waters. A subsequent emergence of O3:K6 strains was detected in austral Chile in 2004. The origin of these 1997 and 2004 population radiations has not yet been conclusively determined. Multiple loci VNTR analysis using seven polymorphic loci was carried out with a number of representative strains from Asia, Peru and Chile to determine their genetic characteristics and population structure. Asian and Chilean subpopulations were the most genetically distant groups with an intermediate subpopulation in Peru. Population structure inferred from a minimum‐spanning tree and Bayesian analysis divided the populations into two genetically distinct groups, consistent with the epidemic dynamics of the O3:K6 clone in South America. One group comprised strains from the original Asiatic population and strains arriving in Peru and Chile in 1997. The second group included the remaining Peruvian Strains and Chilean strains obtained from Puerto Montt in 2004. The analysis of the arrival of the O3:K6 clone at the Pacific coasts of South America has provided novel insights linking the origin of the invasion in 1997 to Asian populations and describing the successful establishment of the O3:K6 populations, first in Peru and subsequently in the South of Chile owing to a possible radiation of Peruvian populations.     

Genetic diversity, recombination and cryptic clades in Pseudomonas viridiflava infecting natural populations of Arabidopsis thaliana

Species-level genetic diversity and recombination in bacterial pathogens of wild plant populations have been nearly unexplored. Pseudomonas viridiflava is a common natural bacterial pathogen of Arabidopsis thaliana, for which pathogen defense genes and mechanisms are becoming increasing well known. The genetic variation contained within a worldwide sample of P. viridiflava collected from wild populations of A. thaliana was investigated using five genomic sequence fragments totaling 2.3 kb. Two distinct and deeply diverged clades were found within the P. viridiflava sample and in close proximity in multiple populations, each genetically diverse with synonymous variation as high as 9.3% in one of these clades. Within clades, there is evidence of frequent recombination within and between each sequenced locus and little geographic differentiation. Isolates from both clades were also found in a small sample of other herbaceous species in Midwest populations, indicating a possibly broad host range for P. viridiflava. The high levels of genetic variation and recombination together with a lack of geographic differentiation in this pathogen distinguish it from other bacterial plant pathogens for which intraspecific variation has been examined.     

Epidemic enhancement in partially immune populations

We observe that a pathogen introduced into a population containing individuals with acquired immunity can result in an epidemic longer in duration and/or larger in size than if the pathogen were introduced into a naive population. We call this phenomenon "epidemic enhancement," and use simple dynamical models to show that it is a realistic scenario within the parameter ranges of many common infectious diseases. This finding implies that repeated pathogen introduction or intermediate levels of vaccine coverage can lead to pathogen persistence in populations where extinction would otherwise be expected.     

Natural transformation increases the rate of adaptation in the human pathogen Helicobacter pylori

Gene exchange between individuals can lead to profound evolutionary effects at both the genomic and population levels. These effects have sparked widespread interest in examining the specific adaptive benefits of recombination. Although this work has primarily focused on the benefits of sex in eukaryotes, it is assumed that similar benefits of genetic exchange apply across eukaryotes and prokaryotes. Here we report a direct test of this assumption using the naturally transformable human gastric pathogen Helicobacter pylori as a model organism. We show that genetic exchange accelerates adaptation to a novel laboratory environment within bacterial populations and that a general adaptive advantage exists for naturally transformable strains when transfer occurs among conspecific backgrounds. This finding demonstrates that there are generalized benefits to adaptation in both eukaryotes and prokaryotes even though the underlying processes are mechanistically different.     

The detection and measurement of recombination from sequence data.

There are two types of recombination that we may wish to detect: rare recombinants between members of different populations or species and repeated recombination within a population. Methods appropriate in the former context are inappropriate in the latter because they depend on recognizing the existence of runs of nucleotides with similar ancestry. If recombination is sufficiently frequent, no such runs will be present. Several methods, including the homoplasy test and the incompatibility test, are described that are appropriate for detecting repeated recombination and for measuring its importance, relative to mutation, in causing genetic change. The sensitivity of these tests is investigated by simulating populations with varying frequencies of mutation and recombination and calculating the various statistics on samples.     

Field experiments on sugar-beet powdery mildew, Erysiphe betae

Sugar-beet powdery mildew, Erysiphe betae appeared in East Anglia in late July 1976 and became wide-spread in eastern England during August and September but was scarce in crops in the north and west of England. Fentinhydroxide, sulphur, benomyl and ethirimol controlled the disease but benomyl applied once only and ethirimol were less effective than the other materials. In heavily infected crops two sulphur sprays, the first applied at the onset of the mildew attack, increased sugar yield by 13%. A single early sulphur spray increased yield on average by 9% giving a return of six to seven times the cost of treatment. When the first spray was delayed, mildew control was less effective.     

Evolution of resistance against powdery mildew in winter wheat populations conducted under dynamic management. I – Is specific seedling resistance selected?

Dynamic management has been proposed as a complementary strategy to gene banks for the conservation of genetic resources. The evolution of frequencies of genes for specific resistance towards powdery mildew (caused by Blumeria graminis f. sp. tritici) in populations of a French network for dynamic management of bread wheat genetic resources was investigated after 10 years of multiplication without human selection. The objective was to determine whether specific resistance gene diversity was maintained in the populations and whether any changes could be attributed to selection due to pathogen pressure. Seven populations, originating from four of the network sites, were characterized and compared to the initial population for six specific resistance gene frequencies detected by nine Blumeria graminis f. sp. tritici isolates. Diversity decreased at the population level, but because of a strong differentiation between the populations, this diversity was maintained at the network level. The comparison of Fst parameters estimated on neutral markers (RFLP) and on resistance gene data revealed that in two of the populations specific resistance genes had been selected by pathogen pressure, whereas evolution in two other populations seemed to be the result of genetic drift. For the three last populations, conclusions were less clear, as one had probably experienced a strong bottleneck and the other two presented intermediate Fst values. A dynamic management network with sites contrasted for pathogen pressure, allowing genetic drift in some populations and selection in others, appeared, at least on the short term, to be a good tool for maintaining the diversity of genes for specific resistance to powdery mildew. Received: 15 December 1999 / Accepted: 30 December 1999     

Productive Infection of Bacillus subtilis 168, with Bacteriophage SP-10, Dependent upon Inducing Treatments

Strains of Bacillus that harbor defective phage PBSX were found to be insensitive to SP-10(C), although the phage adsorbed to these insensitive strains. Strains that did not carry the phage were sensitive to SP-10(C). B. subtilis 168 ind(-), which can be tranduced by SP-10(C) but is nonpermissive for the phage, was rendered phage-sensitive after treatment with ultraviolet (UV) light or mitomycin C. After induction with UV light, maximal sensitivity to SP-10(C) was obtained at a multiplicity of infection (MOI) of approximately 14; with mitomycin C induction, an MOI of approximately 1.0 was required. Phage maturation in sensitized cells was followed by plating infected streptomycin-sensitive cells in the presence of streptomycin at various stages during phase development. The latent period was estimated at 60 to 75 min. We suggest that the resistance of B. subtilis 168 to SP-10 is controlled, at least in part, by the presence of a defective prophage.     

Overlapping functions of recD, recJ and recN provide evidence of three epistatic groups of genes in Escherichia coli recombination and DNA repair

The recD, recJ and recN genes of Escherichia coli K-12 have been shown to be involved in genetic recombination and DNA repair in this organism. Yet, mutation of any one of these genes does not seem to interfere much with the recovery of recombinants from conjugational crosses. Strains carrying all possible combinations of mutations inactivating these genes were constructed and examined for their recombination proficiency and sensitivity to UV light. The recD recJ and recJ recN double mutants are moderately sensitive to UV light and slightly deficient in recombination. A combination of mutations in all 3 genes produced strains that are very deficient in recombination (50- to 100-fold reduction) and strikingly sensitive to UV light. We conclude that these genes provide overlapping activities that compensate for one another in the single mutants. On the basis of these and other data, recombination genes are classified into 3 epistatic groups that define activities which function pre-synaptically or post-synaptically to promote genetic exchanges catalysed by RecA.     

Xylitol and D-arabitol toxicities due to derepressed fructose, galactitol, and sorbitol phosphotransferases of Escherichia coli.

d-Arabitol was observed to be toxic to many laboratory strains of Escherichia coli K-12, and xylitol was found to be toxic to an existing E. coli C mutant strain. Fructose-specific components of the phosphoenolpyruvate:sugar phosphotransferase system are required for xylitol toxicity. Selection for xylitol resistance results in Fru(-) strains blocked in fructose phosphotransferase. Introduction of the ptsF or ptsI mutation into a xylitol-sensitive strain eliminates sensitivity. [(14)C]fructose uptake experiments imply that the mutation to xylitol sensitivity, which is co-transducible with ara and leu, results in derepression of normally inducible fructose phosphotransferase. Wild-type strains also become xylitol sensitive if induced by (and then removed from) fructose. Xylitol toxicity is prevented by fructose in both wild-type and mutant strains. Circumstances causing xylitol, a new food additive, to become toxic to an otherwise insensitive wild-type organism have not been reported previously. The d-arabitol-sensitive laboratory strains are galactitol (dulcitol) utilizers, although most other strains are not. Selection for d-arabitol resistance results in Gat(-) strains blocked in a constitutive galactitol-specific component of the phosphotransferase system. A mutation causing d-arabitol sensitivity occurred many years ago in AB284, the parent of AB311, AB312, AB313, and many other strains. d-Arabitol sensitivity also occurs in sorbitol-constitutive strains and is shown, like the previous two instances of pentitol toxicities, to result from a constitutive phosphotransferase, which is blocked in mutants selected for resistance.