Pathogenicity and virulence of south African isolates of Venturia inaequalis

European Journal of Plant Pathology - Many abiotic and biotic factors can negatively affect the production and productivity of coconut cropping systems. The incidence of Lasiodiplodia species in...     

Localized adhesion of nongerminated Venturia inaequalis conidia to leaves and artificial surfaces

Adhesion to the host surface is the first step for successful plant pathogen development and has been reported to be associated with both passive and active processes. For conidia of Venturia inaequalis, which depend on leaf wetness for germination, this process has not yet been described. Conidia of V. inaequalis adhered to wet hydrophobic surfaces immediately after contact to the surface, hours before initiation of germination. Attachment of nongerminated conidia was much better on hydrophobic surfaces, such as apple leaves and polystyrene, than on hydrophilic glass. Conidia released adhesive material localized in a droplet named spore tip glue (STG) at the spore apex which interacted with a contact surface only when water was present. Histochemical investigations indicated the presence of proteins and carbohydrates in STG, lectin labeling the presence of beta-galactose and N-acetylglucosaminyl residues. Transmission electron microscopy revealed two phases in the STG at the tip of dry mature conidia; as STG was present on the outer side of the intact fungal cell wall its formation should be associated with the secretion of glue through pores of the conidial wall. Surface-active substances affected the adhesion of conidia to hydrophobic surfaces stressing the importance of hydrophobic interactions. The use of protein biosynthesis inhibitors did not affect adhesion of conidia indicating that the adhesive material was preformed. It is concluded that the coincidence of STG, contact to a hydrophobic surface, and free water are essential for the adhesion of V. inaequalis conidia.     

Genetic Diversity of Venturia inaequalis Across Europe

The genetic diversity of eleven populations of Venturia inaequalis from five European countries was calculated based on the allele frequencies of 18 random amplified polymorphic DNA markers and the internal transcribed spacer region of the ribosomal DNA. Diversity within each population (H_S) was high with values ranging from 0.26 to 0.33. Average differentiation among populations (G_ST) was 0.11 and populations were isolated by distance (r²=0.50, P<0.01). This indicates that extensive short-distance gene flow occurs in Europe and that dispersal over longer distances also appears to occur frequently enough to prevent differentiation due to genetic drift. We had expected more pronounced differentiation between populations north and south of the Alps, because V. inaequalis is primarily dispersed by rain splashing and wind distribution of leaf litter, and because the Alps might form a barrier to dispersal. However, our results indicate that human-mediated gene flow due to transport of spores or infected plant material has probably occurred.     

Phenotypic instability of flusilazole sensitivity in Venturia inaequalis

Sensitivities of 16 monoconidial isolates of Venturia inaequalis to the sterol demethylation inhibitor flusilazole were found to be stable, when cultures which had been successively transferred to inhibitor-free nutrient agar and maintained at 20 C were retested 6 months after isolation. In contrast, with storage of these isolates at 2 C for 7 additional months, phenotypic sensitivities were subject to change. A high degree of phenotypic instability was also observed for 77 representative strains which had been subcultured for 10 months. In both cases, the majority of strains became more sensitive to flusilazole. The frequency and magnitude of these changes were most pronounced for strains with moderate levels of resistance. More than 70% of the strains investigated were collected from wild-type populations, suggesting that the reversion of resistant strains was not related to a process of unstable adaptation but to a high degree of plasticity in the genetic control of sensitivities to sterol demethylation inhibitors.     

Differential responses of germinating Venturia inaequalis conidia to kresoxim-methyl

A new class of agricultural fungicides derived from strobilurins act as respiration inhibitors by binding to mitochondrial cytochrome b. The effects of the strobilurin, kresoxim-methyl, on conidia germination, mycelial growth and the protection of apple leaves from scab development were investigated for two isolates of Venturia inaequalis randomly selected from a culture collection. Inhibition of mycelial growth required relatively high doses of kresoxim-methyl (ED₅₀=1 μg ml^-1) for both isolates. In comparison, germination of conidia was highly sensitive for one of the isolates (ED₅₀=0·005 μg ml^-1), while the level of inhibition achieved for the second isolate was 60-fold less (ED₅₀=0·3 μg ml^-1). As deduced from identical sequences of cytochrome b cDNAs prepared from both isolates, the different responses of germinating conidia to kresoxim-methyl were not caused by differences in the sequence of cytochrome b as the target site for strobilurin action. Strong synergistic effects of salicylhydroxamic acid on kresoxim-methyl inhibitory potency suggested that the reduced kresoxim-methyl sensitivity observed for germinating conidia was caused by interference of the alternative respiratory pathway with inhibitor action. However, this mechanism of target site circumvention in germinating conidia had no adverse effects on the protection of apple leaves from scab infection by kresoxim-methyl. © 1998 Society of Chemical Industry     

Germinability of Venturia inaequalis Conidia Exposed to Sunlight

ABSTRACT Detached conidia of Venturia inaequalis were exposed simultaneously outdoors to direct sunlight or in an adjacent ventilated enclosure in darkness for periods ranging from 0.5 to 16.5 h. In addition, conidia were either exposed or not exposed to UV light (254 nm) from 1 to 64 min in the laboratory. After exposure, conidia were placed on water agar in closed petri dishes and allowed 24 h to germinate. Germinability of conidia in the laboratory was reduced up to 95% by doses of UV(254) of 8.1 to 10.8 kJ m(-2).Germinability of conidia in the field was reduced up to 95% by doses of UV-B of about 44 kJ m(-2). A significant percentage of conidia survived radiation doses equivalent to 12 h in full sunlight.     

Characterization of Venturia inaequalis pathogenicity on leaf discs of apple trees

Characterization of pathogenicity on whole plants is required to study host-pathogen interactions between Malus × domestica and Venturia inaequalis. We studied the reliability of an in vitro test of pathogenicity on leaf discs. Three strains of V. inaequalis (races 1, 6 and an English race) were inoculated in vitro onto a range of 16 Malus sp. clones including susceptible and resistant clones. The results were compared to those previously obtained in vivo. Resistant clones contained the main major known genes, i.e. Va, Vb, Vbj, Vf, Vg, Vm and Vr. Scab severity and the sporulation of the fungus were assessed 21 days after the inoculation date. The results indicated that it was possible to reproduce incompatible and compatible situations in vitro. A null severity corresponded to the avirulence of the strain for the clone considered. The resistance given by the Vb, Vbj, Vf, Vg, Vm and Vr genes were expressed in vitro. Only the clone carrying the Va gene and inoculated with the race 6 strain presented a compatible situation which was inconsistent with the observations on the whole plant. Improving this test will facilitate studies on the pathogenicity of V. inaequalis populations in relation to resistance genes of the host expressed in vitro as well as its genetic determinism.     

A biochemical mechanism of natural resistance of apple to Venturia inaequalis

Samenvatting In cultuurfiltraten vanVenturia inaequalis, de verwekker van appelschurft, werden stoffen gevonden die toxisch zijn voor blad van appelrassen welke tegen dit physio resistent zijn. Blad van vatbare rassen is ongevoelig. De resistentie komt tot stand doordat deze toxische werking in het blad de vorming veroorzaakt van fungicide oxydatie-producten van phloridzine. Op grond van deze waarnemingen kan een mogelijke interpretatie van de gen-om-gen relatie bij deze plant-parasietverhouding worden opgesteld.     

A dynamic model simulating infection of apple leaves by Venturia inaequalis

A new dynamic model of the infection of apple leaves by Venturia inaequalis is described. The model begins with the release of spores by rain and incorporates the effect of light on the discharge of ascospores from pseudothecia. The model then simulates infection through the sub-processes of germination, appressorium formation and penetration, separately for ascospores and conidia landed concurrently on wet leaves. The rate of the infection process is determined using different equations for ascospores and conidia. Spore mortality when leaves dry is determined by the stage of infection and RH in the dry period. The infection process is driven by surface wetness, temperature and RH. The progress of each infection period is measured as infection efficiency (IE), namely the percentage of landed spores which have penetrated and thereby infected leaves. The final IE quantifies the favourability of weather in each infection period. In orchard tests in each of three years, the new model detected crucial infection periods in spring and early summer which accounted for outbreaks of leaf scab. These periods were not detected by a static model based on Mills'criteria. The models performed similarly in detecting infection periods later in summer.     

A model simulating deposition of Venturia inaequalis ascospores on apple trees*

Based on existing physical theories and models, a dynamic model estimating the concentration of Venturia inaequalis ascospores in the orchard air and their deposition on apple leaves was elaborated. The model produces two main outputs: number of ascospores deposited per leaf and proportion of ascospores discharged from pseudothecia deposited onto the leaves. The model has a relatively simple structure, and computations are based on few algorithms, which are implemented on an electronic data sheet of common use. Nevertheless, it preserves the accuracy of more complex physical models reasonably well. The model includes the effect of meteorological conditions and horticultural characteristics, and thus provides information for each type of orchard. Few input variables are required: wind speed and rainfall rate can be measured in standard meteorological stations; horticultural characteristics of the orchard can be determined for each type of orchard. The model produces conservative estimates of ascospore deposition, because it assumes a complete retention of the spores deposited by rainfall and does not consider either deposition on stems and flowers or the spatial distribution of plant surfaces. After further validation under orchard conditions, the model will be used to obtain better estimates of scab infection risk in current scab control strategies.     

Negative cross-resistance to N-phenylanilines in benzimidazole- resistant strains of Botrytis cinerea,Venturia nashicola and Venturia inaequalis

A new series of compounds with high contra-selective activity against benomyl-resistant fungal strains was found among ring-substituted N-phenyl-anilines. Hydrophobic substitution in one of the benzene rings, together with the secondary amine character of the molecule, are important factors for high fungitoxicity. The sensitivities of benomyl-resistant isolates to some representatives of the N-phenylanilines equal or even surpass their sensitivity to diethofencarb or MDPC. The negative cross-resistance with benzimdazole fungicides is valid for strains of Botrytis cinerea, Venturia nashicola and Venturia inaequalis. The strains of B. cinerea with double insensitivity to benzimidazoles and phenyl-carbamates are insensitive also to N-phenylanilines. However, the similarly double-insensitive strain of V. nashicola was found to be just as sensitive to the N-phenylanilines tested as the strain resistant only to benzimidazoles. The latter result revealed an important bonus compared with diethofencarb. Preventive application of one of the most active representatives of the N-phenylanilines to young cucumber plants was effective against infection with benomyl-resistant isolates of B. cinerea. Moderate apical translocation after root-dipping was also observed.     

Acquisition of Resistance to Sterol Demethylation Inhibitors by Populations of Venturia inaequalis

ABSTRACT Acquisition of resistance to sterol demethylation inhibitors (DMIs) by populations of Venturia inaequalis was investigated using a microscopical method developed by C. Siebels and K. Mendgen. Microscopical analysis of conidiophore formation enabled the earlier detection of resistance and a clearer distinction between DMI-resistant and DMI-sensitive populations than other in vivo methods commonly used to analyze inhibitory effects of fungicides. In addition, because observations were made on the level of individuals, quantitative measures of the composition of conidial populations were obtained. The development of DMI sensitivity was followed over a period of 3 years in control apple orchards that had never been treated with fungicides and in orchards with DMI history. The 50% effective dose values determined by microscopical evaluation of conidio-phore development for untreated populations revealed the baseline sensitivities of 0.3, 0.96, 0.09, 1.22, and 1.92 mg/liter for flusilazole, fenarimol, difenoconazole, tebuconazole, and pyrifenox, respectively. As compared with the baseline sensitivity, all populations with DMI history showed significant resistance to flusilazole. A strong nonlinear correlation (R = 0.96) was found between the resistance factors and the sum of all DMI treatments of the 3 years before taking the sample. According to this correlation, resistance can be expected in all apple orchards of the fruit-growing area along Lake Constance, Germany, in which more than two DMI treatments per season have been applied. Due to cross-resistance, the recently introduced DMI fungicides difenoconazole, tebuconazole, and pyrifenox did not allow the control of V. inaequalis populations resistant to flusilazole.     

Mutations in Cyp51 of Venturia inaequalis and their effects on DMI sensitivity

Journal of Plant Diseases and Protection - DMIs (sterol demethylation inhibitors) provide an important mode of action for control of Venturia inaequalis. They target the enzyme 14a-demethylase,...     

Simulation of Epidemics Caused by Venturia inaequalis (Cooke) Aderh.1

We discuss simulation of apple scab epidemics based on analog computer models, multivariate regression analyses, and systems analyses. Details of underlying models and their scope for applications are emphasized. Monomolecular growth functions by Bertalanffy and Gompertz in analog computers permit fast simulation of disease progress curves. The models derived from multivariate analyses of field experiments on apple scab epidemics simulate closely the changes in infection rates. EPIVEN, our comprehensive and self–generating simulator of apple scab epidemics is reviewed and compared to a reduced model, also based on elements of the system “apple scab epidemic”.     

Identification of Microsatellite Markers and Their Application to Population Genetics of Venturia inaequalis

ABSTRACT Microsatellite markers of Venturia inaequalis were developed using genomic libraries of V. inaequalis enriched for the simple sequence repeats (TC)(n) and (AAC)(n). Seven markers, three with (TC)(n) repeats and four with (AAC)(n) repeats, were selected for the analyses of 350 isolates of V. inaequalis collected from 11 sites in Europe. Polymorphism in the (TC)(n) repeats was higher than in the (AAC)(n) repeats. Nei's expected genetic diversity (H(E)) varied between 0.52 and 0.96 for the microsatellites containing (TC)(n) stretches and between 0.09 and 0.36 for the microsatellites containing (AAC)(n) stretches. Within-population diversity (H(S)) was very high with values ranging from 0.28 to 0.49, whereas differentiation among all European populations (G(ST)) was low with an average of 0.07. In the population from Ahrensburg (northern Germany) where isolates were mainly collected from apple varieties carrying the Vf gene, usually resistant to V. inaequalis, we showed a bottleneck effect with reduced diversity and loss of alleles. The great advantages of microsatellite markers over random amplified polymorphic DNA and polymerase chain reaction-restriction fragment length polymorphism markers are their high specificity, high polymorphism, good reproducibility, and unambiguous scorability.     

Genetics of Host-Pathogen Relationships Between Venturia inaequalis Races 6 and 7 and Malus Species

ABSTRACT Resistance to scab originating from Malus floribunda clone 821 is the most widely form of resistance used in apple breeding programs. A dominant gene, named Vf, was introgressed from this clone into recent cultivars, although the genetic determinants of the resistance of M. floribunda 821 are apparently more complex than a single gene. The appearance of new races overcoming the resistance of cultivars with the Vf gene, the parental clone, or both made it possible to undertake a genetic analysis of host-pathogen interactions. The segregation of resistance in progenies of crosses from 'Golden Delicious' x M. floribunda 821 and 'Golden Delicious' x 'Idared' into five strains of Venturia inaequalis-races 1 (strains 104, 1093, and 301), 6 (strain 302), and 7 (strain 1066)-demonstrated the existence of a second dominant gene in M. floribunda 821. This gene, independent of Vf, was named Vfh because it seemed to induce a hypersensitive reaction. The results obtained with strain 1066, virulent to M. floribunda 821, allowed identification of another dominant gene, Vg, responsible for the resistance of 'Golden Delicious' to this strain. Vg is also carried by 'Florina', which was selected for its Vf resistance. The pathogenicity of a progeny originating from a cross between V. inaequalis strains 1066 and 301, characterized in vitro on leaf disks of differential genotypes, revealed two independent avirulence genes involved in the pathogenicity toward the Vg and Vf genes, respectively. These avirulence genes were named Avr Vg and Avr Vf. The host-pathogen interactions detected are consistent with a gene-for-gene relationship.     

Virulence characteristics of apple scab (Venturia inaequalis) isolates from monoculture and mixed orchards

Isolates of Venturia inaequalis were sampled from monoculture and mixed orchards of three apple cultivars: Bramley, Cox and Worcester. In addition, single-ascospore progeny isolates were obtained from three crosses between pairs of isolates originating from the three cultivars in monoculture orchards. These field isolates and single-ascospore progenies were inoculated onto each of the three cultivars in a glasshouse. The patterns of infection showed that all three cultivars, commonly regarded as susceptible to scab, contained some specific resistance factors and that scab isolates from both mixed and monoculture orchards appeared to have co-evolved with cultivars. A much higher proportion of isolates from cv. Worcester in the mixed orchard were unable to infect any of the three cultivars than isolates from any other combination of cultivar and orchard type, but there was no other difference between isolates from mixed vs. monoculture orchards. Many isolates could infect both Bramley and Cox, or Cox and Worcester; but only a single isolate could infect both Bramley and Worcester. Results from the testing of 61 single-ascospore progeny isolates suggested that virulence towards Bramley, Cox and Worcester was controlled by at least one, two and three factors, respectively. Moreover, the exact nature of the interactions between these factors in determining virulence depended on the particular pair of isolates concerned.     

Evidence for the Predisposition of Fungicide-Resistant Isolates of Venturia inaequalis to a Preferential Selection for Resistance to Other Fungicides

In the United States, populations of the apple scab pathogen Venturia inaequalis have progressed through three consecutive rounds of fungicide resistance development, first to dodine, then to the benzimidazoles, and most recently to the sterol demethylation inhibitors (DMIs). Analysis of extensive monitoring data have to date provided no indication of detectable cross-resistance or partial cross-resistance of V. inaequalis populations to the three unrelated classes of fungicides prior to the selection of resistant subpopulations. However, in this study, resistance to both benomyl and DMIs developed to significantly higher frequencies within the previously established dodine-resistant population than in the population sensitive to dodine. Accelerated selection of phenotypes double resistant to dodine and the DMI fenarimol was apparent over the course of distinct seasons of apple scab management with either dodine or fenarimol. The data provide evidence for an accelerated speed of resistance development among phenotypes of V. inaequalis already resistant to an unrelated fungicide. This finding represents a departure from the previous model, which assumed entirely independent rounds of resistance developments. The data indicate that phenotypes of V. inaequalis might not only be selected for the trait of fungicide resistance but also for traits allowing a more flexible response to changes in the environment where they compete.     

Control of apple scab (Venturia inaequalis) with bicarbonate salts under controlled environment

Journal of Plant Diseases and Protection - The effectiveness of potassium bicarbonate against Venturi a inaequalis, the cause of apple scab, was studied. In vitro experiments with sodium, ammonium...