Arne Stensvand

Induced Resistance as a Possible Means to Control Diseases of Strawberry Caused by Phytophthora spp.

Two putative elicitors of disease resistance (acibenzolar-S-methyl and chitosan) were tested for their effect on crown rot (Phytophthora cactorum) in strawberry. The effect of both compounds was enhanced when the time between treatment and inoculation was prolonged from 2 to 20 days. There were no significant differences between treatments when the concentration of acibenzolar-S-methyl was increased from 10 to 1,000 μg a.i./plant. The lowest tested concentrations of chitosan (10 and 50 μg a.i./plant) resulted in a lower disease score compared with the highest concentrations (250 or 1,000 μg a.i./plant). There were no differences in disease score between treatment with fosetyl-Al, acibenzolar-S-methyl, or chitosan when applied 5 or 15 days before inoculation. The effect of acibenzolar-S-methyl and chitosan also was tested against P. fragariae var. fragariae in alpine strawberry (Fragaria vesca var. alpina cv. Alexandria). Chitosan had no effect, whereas fosetyl-Al and all treatments with acibenzolar-S-methyl (50 or 250 μg a.i./plant; 5, 10, 20, or 40 days before inoculation) reduced the severity of the disease. There were no significant differences between acibenzolar-S-methyl and fosetyl-Al when applied at the same time. Acibenzolar-S-methyl and chitosan at concentrations of 0.5, 5, 50, and 500 μg a.i. ml-1 in V8 juice agar were tested for possible effects on P. cactorum and P. fragariae var. fragariae in vitro. Only chitosan at concentrations of 50 and 500 μg a.i. ml-1 had a growth-retarding effect on P. cactorum. Both acibenzolar-S-methyl and chitosan at a concentration of 500 μg a.i. ml-1 reduced the growth rate of P. fragariae var. fragariae.

Antagonism of Nutrient-Activated Conidia of Trichoderma harzianum (atroviride) P1 Against Botrytis cinerea.

ABSTRACT The effect of preliminary nutrient activation on the ability of conidia of the antagonist Trichoderma harzianum (atroviride) P1 to suppress Botrytis cinerea was investigated in laboratory, greenhouse, and field trials. Preliminary nutrient activation at 21 degrees C accelerated subsequent germination of the antagonist at temperatures from 9 to 21 degrees C; at >/=18 degrees C, the germination time of preactivated T. harzianum P1 conidia did not differ significantly from that of B. cinerea. When coinoculated with B. cinerea, concentrated inocula of preactivated but ungerminated T. harzianum P1 conidia reduced in vitro germination of the pathogen by >/=87% at 12 to 25 degrees C; initially quiescent conidia achieved this level of suppression only at 25 degrees C. Application of quiescent T. harzianum P1 conidia to detached strawberry flowers in moist chambers reduced infection by B. cinerea by >/=85% at 24 degrees C, but only by 35% at 12 degrees C. Preactivated conidia reduced infection by >/=60% at 12 degrees C. Both quiescent and preactivated conidia significantly reduced latent infection in greenhouse-grown strawberries at a mean temperature of 19 degrees C, whereas only preactivated conidia were effective in the field at a mean temperature of 14 degrees C on the day of treatment application. An antagonistic mechanism based on initiation of germination in sufficiently concentrated inocula suggests that at suboptimal temperatures the efficacy of Trichoderma antagonists might be improved by conidia activation prior to application.

Influence of Light, Relative Humidity, and Maturity of Populations on Discharge of Ascospores of Venturia inaequalis

ABSTRACT Ascospore release in 20 populations of Venturia inaequalis was generally suppressed in wind tunnel tests during darkness and simulated rain, but the following relieved this suppression: (i) exposure to low relative humidity during simulated rain and (ii) protracted incubation of leaf samples and the consequent senescence of the pathogen population. No counterpart to (i) was observed under orchard conditions. Although V. inaequalis also released a high percentage of ascospores during darkness in field studies under simulated rain late in the season of ascospore release, this phenomenon has not been reported for natural rain events. A threshold value of 0.5 muW/cm(2) at 725 nm was identified as the minimum stimulatory light intensity. Ascospore release increased with increasing light intensity from 0.5 to 5.2 muW/cm(2) at 725 nm. There was also an intrinsic increase in ascospore release as duration of rain increased. In orchards, the combined impact of both processes is probably responsible for a delay in reaching peak ascospore release at several hours after sunrise. Ascospore release during darkness will generally constitute a small proportion of the total available supply of primary inoculum. Significant ascospore release, and therefore infection periods, can be assumed to begin shortly after sunrise, when rain begins at night in orchards with low potential ascospore dose (PAD). A PAD level of 1,000 ascospores per m(2) of orchard floor per season is suggested as a threshold, above which the night-released ascospores should not be ignored.

Specific Light-Emitting Diodes Can Suppress Sporulation of Podosphaera pannosa on Greenhouse Roses

When rose plants bearing colonies of Podosphaera pannosa were placed in a wind tunnel, the number of conidia trapped was directly proportional to intensity of daylight-balanced (white) light from 5 to 150 μmol m-2 s-1. Illumination of samples using blue (420 to 520 nm) light-emitting diodes (LEDs) increased the number of conidia trapped by a factor of approximately 2.7 over white light but germination of conidia under blue light was reduced by approximately 16.5% compared with conidia germination under white light. The number of conidia trapped under far-red (>685 nm) LEDs was approximately 4.7 times higher than in white light, and 13.3 times higher than under red (575 to 675 nm) LEDs, and germination was not induced compared with white light. When mildewed plants were exposed to cycles of 18 h of white light followed by 6 h of blue, red, far-red light, or darkness, light from the red LEDs reduced the number of conidia trapped by approximately 88% compared with darkness or far-red light. Interrupting the above dark period with 1 h of light from red LEDs also reduced the number of conidia trapped, while a 1-h period of light from far-red following the 1 h of light from red LEDs nullified the suppressive effect of red light. Our results indicate that brief exposure to red light during the dark interval may be as effective as continuous illumination in suppressing powdery mildew in greenhouse rose plant (Rosa × hybrida).

Discharge and Dissemination of Ascospores by Venturia inaequalis During Dew

Abundant airborne ascospores of the apple scab pathogen (Venturia inaequalis) have never before been observed during periods of dew. We studied ascospore release in V. inaequalis in two orchards in southeastern Norway using Burkard 7-day volumetric spore traps. At Ås in 1990, 1992, and 1997, and at Svelvik in 1992, a total of 14.8, 1.4, 0.27, and 26.9%, respectively, of the seasons total spore release was trapped during periods of dew. Dew followed by spore release was observed 22 days at the two locations. During one night with dew at Ås in 1990 and two nights with dew at Svelvik in 1992, approximately 13 and 20%, respectively, of the seasons total spore numbers were observed. High numbers of spores were trapped prior to sunrise, and on an average, 48.4% of the spores were trapped prior to 0400 in the morning. Episodes in which more than 1% of the seasons inoculum was released during dew occurred around bloom of apple, which is the peak period for ascospore discharge, and followed more than 2 days of fair weather (clear, warm days and cool, humid nights). The ordinary suppression of ascospore release in V. inaequalis during darkness has been overcome in previous studies under laboratory conditions when protracted periods favorable for ascospore maturity occur without opportunity for ascospore discharge. This is the first confirmed report of relatively large (>10% of the seasons total inoculum) numbers of airborne ascospores in orchards during dew. The sequential occurrence of specific weather conditions, for example (i) fair-weather days, (ii) cool nights with abundant dew formation, (iii) significant release and dispersal of airborne ascospores, and (iv) poor drying conditions or additional hours of leaf wetness due to fog or rain, would be required for dew-released ascospores to constitute a threat of infection. Absent the foregoing, release during dew is more likely to deplete the ascospore supply with no consequent increase in the overall risk of disease.

Initiation, development, and survival of cleistothecia of Podosphaera aphanis and their role in the epidemiology of strawberry powdery mildew.

A collection of four clonal isolates of Podosphaera aphanis was heterothallic and was composed of two mutually exclusive mating types. Cleistothecial initials approximately 20 to 30 microm in diameter were observed within 7 to 14 days after pairing of compatible isolates and developed into morphologically mature ascocarps within 4 weeks after initiation on both potted plants maintained in isolation and in field plantings in New York State and southern Norway. Ascospores progressed through a lengthy maturation process over winter, during which (i) the conspicuous epiplasm of the ascus was absorbed; (ii) the osmotic potential of the ascospore cytoplasm increased, resulting in bursting of prematurely freed spores in water; and, finally, (iii) resulting in the development of physiologically mature, germinable, and infectious ascospores. Release of overwintered ascospores from field collections was coincident with renewed plant growth in spring. Overwintered cleistothecia readily dehisced when wetted and released ascospores onto glass slides, detached strawberry leaves, and leaves of potted plants. Plant material exposed to discharged ascospores developed macroscopically visible mildew colonies within 7 to 10 days while noninoculated controls remained mildew free. Scanning electron and light microscopy revealed that cleistothecia of P. aphanis were enmeshed within a dense mat of hyphae on the persistent leaves of field-grown strawberry plants and were highly resistant to removal by rain while these leaves remained alive. In contrast, morphologically mature cleistothecia on leaves of nine deciduous perennial plant species were readily detached by simulated rain and seemed adapted for passive dispersal by rain to other substrates. Contrary to many previous reports, cleistothecia appear to be a functional source of primary inoculum for strawberry powdery mildew. Furthermore, they differ substantially from cleistothecia of powdery mildews of many deciduous perennial plants in their propensity to remain attached to the persistent leaves of their host during the intercrop period.

A comparison of methods used to estimate the maturity and release of ascospores of Venturia inaequalis

Maturation and release of ascospores of Venturia inaequalis were assessed at Geneva and Highland, NY, and at Durham, NH, by microscopic examination of crushed pseudothecia excised from infected apple leaves that were collected weekly from orchards (squash mounts) in 14 siteyear combinations. Airborne ascospore dose was monitored at each location in each year of the study by volumetric spore traps. Additional laboratory assessments were made at Geneva to quantify release from infected leaf segments upon wetting (discharge tests). Finally, ascospore maturity was estimated for each location using a degree-day model developed in an earlier study. Ascospore maturation and release determined by squash mounts and discharge tests lagged significantly behind cumulative ascospore release as measured by volumetric spore traps in the field. The mean date of 98% ascospore discharge as determined by squash mounts or discharge tests occurred from 23 to 28 days after the mean date on which 98% cumulative ascospore release had been detected by volumetric traps. In contrast, cumulative ascospore maturity estimated by the degree-day model was highly correlated (r2 = 0.82) with observed cumulative ascospore release as monitored by the volumetric traps. Although large differences between predicted maturity and observed discharge were common during the exponential phase of ascospore development, the date of 98% cumulative ascospore maturity predicted by the model was generally within 1 to 9 calendar days of the date of 98% cumulative ascospore recovery in the volumetric traps. Cumulative ascospore discharge as monitored by the volumetric traps always exceeded 98% at 600 degree days (base = 0°C) after green tip. Estimating the relative quantity of primary inoculum indirectly by means of a degree-day model was more closely aligned with observed ascospore release, as measured by volumetric traps, than actual assessments of ascospore maturity and discharge obtained through squash mounts and discharge tests. The degree-day model, therefore, may be a more accurate predictor of ascospore depletion than squash mounts or discharged tests, and has the added advantage that it can be widely applied to generate site-specific estimates of ascospore maturity for any location where daily temperature data are available.

Ontogenic resistance of leaves and fruit, and how leaf folding influences the distribution of powdery mildew on strawberry plants colonized by Podosphaera aphanis.

Ontogenic or age-related resistance has been noted in many pathosystems but is less often quantified or expressed in a manner that allows the concept to be applied in disease management programs. Preliminary studies indicated that leaves and fruit of three strawberry cultivars rapidly acquired ontogenic resistance to the powdery mildew pathogen, Podosphaera aphanis. In the present study, we quantify the development of ontogenic resistance in controlled inoculations of 10 strawberry cultivars using diverse isolates of P. aphanis in New York and Florida, USA, and in Norway. We report the differential and organ-specific development of ontogenic resistance in the receptacle and externally borne strawberry achenes. We further report that rapid development of ontogenic resistance prior to unfolding of emergent leaves, rather than differential susceptibility of adaxial versus abaxial leaf surfaces, may explain the commonly observed predominance of powdery mildew on the lower leaf surfaces. Susceptibility of leaves and fruit declined exponentially with age. Receptacle tissue of berries inoculated at four phenological stages from bloom to ripe fruit became nearly immune to infection approximately 10 to 15 days after bloom, as fruit transitioned from the early green to the late green or early white stage of berry development, although the achenes remained susceptible for a longer period. Leaves also acquired ontogenic resistance early in their development, and they were highly resistant shortly after unfolding and before the upper surface was fully exposed. No significant difference was found in the susceptibility of the adaxial versus abaxial surfaces. The rapid acquisition of ontogenic resistance by leaves and fruit revealed a narrow window of susceptibility to which management programs might be advantageously adapted.

Suppression of Cucumber Powdery Mildew by Supplemental UV-B Radiation in Greenhouses Can be Augmented or Reduced by Background Radiation Quality

This study demonstrates that the spectral quality of radiation sources applied with ultraviolet-B (UV-B; background radiation) affects the suppression of cucumber powdery mildew (Podosphaera xanthii) by UV-B. Suppression provided by daily UV-B exposure of 1 W/m2 for 10 min was greatest in the presence of red light or by a complete lack of background light, and powdery mildew suppression was least in the presence of ultraviolet-A (UV-A) or blue radiation compared with plants exposed only to 16 h of daily natural light supplemented with high-pressure sodium lamps that supply broad-spectrum radiation with peaks in the yellow-orange region. Exposure of powdery mildew-inoculated plants to supplemental red light without UV-B, beginning at the end of the daylight period, also reduced disease severity; however, supplemental blue light applied in the same fashion had no effect. Daily application of UV-B at 1 W/m2 beginning on the day of inoculation significantly reduced the severity of powdery mildew to 15% compared with 100% severity on control plants. Maximum suppression of powdery mildew was observed following 15 min of exposure to UV-B (1.1% severity compared with 100% severity on control plants) but exposure time had to be limited to 5 to 10 min to reduce phytotoxicity. There was no additional disease suppression when plants were exposed to UV-B beginning 2 days prior to inoculation compared with plants exposed to UV-B beginning on the day of inoculation. UV-B inhibited germination, infection, colony expansion, and sporulation of P. xanthii. The results suggest that efficacy of UV-B treatments, alone or in combination with red light, against P. xanthii can be enhanced by exposure of inoculated plants to these wavelengths of radiation during the night, thereby circumventing the counteracting effects of blue light and UV-A radiation. The effect of UV-B on powdery mildew seemed to be directly upon the pathogen, rather than induced resistance of the host. Night exposure of plants to 5 to 10 min of UV-B at 1 W/m2 and inexpensive, spectral-specific, light-emitting diodes may provide additional tools to suppress powdery mildews of diverse greenhouse crops.

Evaluation of Six Models to Estimate Ascospore Maturation in Venturia pyrina

Estimates of ascospore maturity generated by models developed for Venturia pyrina in Victoria, Australia (NV and SV), Oregon, United States (OR), and Italy (IT) or for V. inaequalis in New Hampshire, United States (NH-1) or modified in Norway (NH-2) were compared with observed field ascospore release of V. pyrina from 21 site-year combinations. The models were also compared with ascospore release data from laboratory assays. In the laboratory assays, the forecasts of the NH-1 and NH-2 models provided the best fit to observed spore release. Under field conditions, the lag phases and slope coefficients of all models differed from those of observed release of ascospores. Identifying the precise time of bud break of pear to initiate degree-day accumulation was problematic at both Australian sites. This resulted in a higher deviance between bud break and first released ascospore compared with the sites in Norway and Belgium. Linear regressions of observed release against forecasted maturity generated similarly high concordance correlation coefficients. However, where differences were noted, they most often favored models that included adjustment for dry periods. The NH-2, IT, and NV models using pooled data also provided the most accurate estimates of 95% ascospore depletion, a key event in many disease management programs.