Influence of environmental conditions on dispersal of Botrytis cinerea conidia and on post-harvest infection of gerbera flowers grown under glass

Dispersal of Botrytis cinerea in a gerbera crop grown in two glasshouses 30 km apart was studied over a period of 18 months, in 1988 and 1989. Conidia were caught in spore traps consisting of agar in petri dishes exposed at different heights in the crop in each glasshouse. No seasonal patterns could be identified in the spore catches, assessed as colonies on the agar traps after incubation. The number of lesions caused by conidial infection of gerbera flowers following incubation, however, showed a distinct pattern. In spring and early summer few lesions were recorded whereas at other times of the year many lesions appeared. In linear regression analysis, variation in numbers of colonies (spore catches) could not be explained by environmental factors recorded during the experiments. Linear regression accounted for 77% and 81% of the variation in the number of lesions on flowers in the two glasshouses, in terms of relative humidity (postively correlated), global radiation outside the glasshouse (negatively correlated) and age of the crop (positively correlated). Despite differences in the systems by which the gerbera crop was produced and in the spore catches, the numbers of lesions on gerbera flowers in the two glasshouses were significantly correlated though not significantly different from each other.     

Horizontal and vertical distribution of airborne conidia of Botrytis cinerea in a gerbera crop grown under glass

The horizontal and vertical distribution of airborne conidia ofBotrytis cinerea in a gerbera crop in two glasshouses (100 m² and 350 m²) was studied during 18 months in 1988 and 1989. Conidia ofB. cinerea were caught in simple spore traps consisting of agar in Petri dishes placed in a regular pattern at three different heights in the glasshouse and counted as colonies, after incubation. Lesions due to conidial infection were counted on gerbera petals. The horizontal and vertical distribution of conidia ofB. cinerea in a gerbera crop grown under glass was fairly uniform in both distinct glass-houses. Conidia ofB. cinerea trapped in a glasshouse can originate from sources inside and outside the glasshouse. No significant interaction was found between location and time for the colony counts and for the log transformed (ln(N+1)) lesion counts. The results of this study suggest that spore trapping at one height and at a limited number of locations and dates is sufficient for efficient monitoring ofB. cinerea in a glasshouse.     

Effects of temperature,vapour pressure deficit and radiation on infectivity of conidia ofBotrytis cinerea and on susceptibility of gerbera petals

The effect of vapour pressure deficit, temperature and radiation on the postharvest susceptibility of gerbera flowers toB. cinerea, on the water relations of gerbera flowers and on the lesion formation after conidial infection ofB. cinerea was studied. The temperature range in whichB. cinerea could germinate and growin vitro is 5–30 °C. In climate chamber experiments flowers had more lesions ofB. cinerea at temperatures of 20 and 25 °C than at 10 and 15°C. At 15, 20 and 25°C the infectivity ofB. cinerea conidia was negatively affected during a storage-period of 7 days. At a vapour pressure deficit (VPD) of 200 Pa significantly more conidia ofB. cinerea were infective than at 800 Pa. At a VPD of 800 Pa the susceptibility of gerbera flowers forB. cinerea was not significantly different than at 200 Pa. High radiation levels in glasshouses in spring and summer negatively influenced the infectivity of conidia ofB. cinerea on the flower surface, but did not affect the susceptibility of gerbera flowers forB. cinerea. In spring and early summer conidia lost their infectivity at high radiation levels, high temperatures and high levels of VPD. In summer gerbera flowers could be more susceptible toB. cinerea because of high temperatures in glasshouses, but the negative effect of radiation on the conidia ofB. cinerea seemed to overrule the temperature effect. Thus, the numbers of lesions in spring and summer can be low compared with the numbers in other seasons, although the numbers ofB. cinerea colonies on spore traps can be high. The effect of temperature on the susceptibility of gerbera flowers can probably be explained by changes of water status in the petals. At higher temperatures the number of lesions and the turgor (=water potential—osmotic potential) in the petals increased. Temperatures <10°C during lesion formation (RH>95% and VPD<50 Pa) had a temporary negative effect on the number of lesions. After 3 days of incubation the numbers of lesions were about equal (30 lesions/cm²) from 5 to 20°C. At 30°C no lesion formation was observed even after 3 days.     

Relations between glasshouse climate and dry weight of petals,epicuticular wax,cuticle, pre-harvest flowering period and susceptibility toBotrytis cinerea of gerbera and rose flowers

Studies were conducted on the effects of seasonal levels of relative humidity, temperature, and total radiation, on dry weight of petals, on fresh weight of epicuticular wax and of cuticle of petals, on numbers ofBotrytis cinerea lesions in petals, and on preharvest flowering periods in gerbera and rose. No temporal relationships or significant correlations were found among dry weight of petals, weight of wax and cuticle of petals, and numbers of lesions on the petals. Temperature, relative humidity and total radiation did not correlate significantly with dry weight of petals, or with fresh weights of wax and cuticle of petals, except for a positive correlation between relative humidity and cuticle weight in the gerbera cultivar Delphi. No relation was found between weight of epicuticular wax and cuticle of petals and susceptibility of gerbera and rose petals toB. cinerea. The thickness of wax and cuticle on flowers did not seem to be an important factor influencing the susceptibility of flowers toB. cinerea. The seasonal pattern in number of lesions produced on the flowers byB. cinerea was related to the effects of relative humidity and radiation on infectivity of conidia of the pathogen on the flower surface but not to the effects on the susceptibility of flowers.     

Effects of Aureobasidium pullulans on numbers of lesions on dwarf bean leaves caused by Alternaria zinniae

Inoculating dwarf bean leaves with spore suspensions ofAureobasidium pullulans one day before or simultaneously with inoculations made withAlternaria zinniae significantly decreased numbers of lesions caused by the latter fungus. Delaying inoculation withA. pullulans until 1 day after that withA. zinniae lessened the effect on lesion numbers. Increasing numbers ofA. pullulans spores progressively decreased numbers ofA. zinniae lesions. Three of five isolates ofA. pullulans decreased lesion numbers caused byA. zinniae. AlthoughA. pullulans restricted the germination ofA. zinniae on dwarf bean leaves, it had no effect when on cherry agar. Few viable propagules ofA. pullulans were recovered from dwarf bean leaves when examined 7 days after application.     

Failure to detect highly resistant strains in dicarboximide resistant field populations of Botrytis cinerea1

Field isolates of Botrytis cinerea with moderate levels of resistance to dicarboximide fungicides (ED50 1.0–4.9 μg ml^?1) and to dicloran were obtained from glasshouses where vinclozolin and iprodione failed to control grey mould. From sensitive and moderatcly-resistant cultures, laboratory isolates were selected on dicarboximide-amended medium, which were highly resistant to these fungicides (ED50 125->3000 μg ml^?1). Conidia of all the resistant isolates germinated well on media amended with 100 μg ml^?1 of the dicarboximides vinclozolin, iprodione, procymidone and myclozolin and with 5 μg ml^?1 of metomeclan. However, the spores of the moderately resistant isolates did not germinate on 100 μg ml^?1 metomeclan while the spores of the highly resistant isolates germinated well. Using media with 100 μg ml^?1 of metomeclan to distinguish between the two phenotypes, no highly resistant strain was detected among 312 resistant samples from five cucumber glasshouses with a high frequency of moderately resistant strains. From air-borne inoculum of five glasshouses with 100% resistant populations, 1604 colonies were recovered on vinclozolin-amended (100 μg ml^?1) medium and none on metomeclan-amended (100 μg ml^?1) medium. It is concluded that strains of B. cinerea highly resistant to dicarboximides are absent from field populations.     

Biological control ofBotrytis cinerea in residues and flowers of Rose (Rosa hybrida)

Microbial isolates from living petals, petal residues and leaf residues of rose, and from laboratory collections, were evaluated for control ofBotrytis cinerea in rose. In leaf residues artificially infested withB. cinerea, isolates of the filamentous fungiGliocladium roseum, FR136 (unidentified) andTrichoderma inhamatum reduced sporulation of the pathogen by >90%, other filamentous fungi were 25–90% effective, and those of yeasts and bacteria were <50% effective. In artificially inoculated petal residues, no microbe reduced sporulation ofB. cinerea by >75%, but isolates ofCladosporium oxysporum and four yeasts were 51–75% effective, and three filamentous fungi, eight yeasts andBacillus subtilis isolates were 26–50% effective. Isolates ofT. inhamatum, C. oxysporum andG. roseum performed best againstB. cinerea among isolates evaluated in leaf residues naturally infested with the pathogen and indigenous microorganisms. Totals of ten isolates of filamentous fungi (includingC. oxysporum andC. cladosporioides), two of yeasts and five ofBacillus subtilis completely prevented lesion production byB. cinerea in detached petals, and a further six isolates of filamentous fungi (includingG. roseum) and six yeasts were 90–99% effective. Isolates ofC. oxysporum, C. cladosporioides andB. subtilis, the most effective microorganisms againstB. cinerea in flower buds, reduced number of lesions in the range of 42–65% compared with 59–89% for à standard fungicide (vinclozolin). It is suggested that application of leading antagonists Jo living rose leaves and flowers should optimize control of inoculum production byB. cinerea when the tissues die. Optimal biocontrol of lesion production in flower buds requires a better understanding of the microenvironment of petals.     

A selective medium for Botrytis cinerea to be used in a spore-trap

A selective medium has been developed for the use in spore-traps to study the dispersion ofB. cinerea on gerbera grown in glasshouses.Samenvatting Een selectief medium voorB. cinerea is, ontwikkeld voor het gebruik in sporevangers. Deze sporevangers werden gebruikt bij het bestuderen van de ontwikkeling vanB. cinerea in gerbera geteeld onder glas.     

Epidemiological aspects of Didymella bryoniae,the cause of stem and fruit rot of cucumber

The survival ofDidymella bryoniae and the incidence of ascospores in glasshouses, outdoors and under controlled conditions were studied. The fungus was able to overwinter in the open as dormant mycelium. Dry and undecomposed crop residues remained a source of infection for more than one year. Moisture and a minimum temperature between 5 and 10°C were needed for fructification. For ascospore release a high relative humidity was not sufficient, the substrate had to be moist during a short period. Ascospores could be trapped throughout day and night both outdoors and in glasshouses, but there was a marked peak during a period of 3 h in the evening. Both on days with and without rain about the same numbers of ascospores were trapped from crop residues in the open. Ascospore release was favoured by watering the plants in the glasshouse. Under controlled conditions the release of ascospores was determined by humidity and not by light or darkness. In a cucumber crop in the glasshouse the first ascospores were trapped at about the same time the first symptoms on the plants appeared. In the glasshouse with introduced diseased plant debris, particularly when the debris became wet when the plants were watered, the disease was more severe and yield was less than in a glasshouse without introduced plant debris. Airborne ascospores may cause the primary infection of a cucumber crop. Therefore, hygienic measures must be taken to eliminate plant debris as source of infection, both in glasshouses and outdoors.     

Epidemiology of Grey Mould in Annual Waiting-bed Production of Strawberry

The epidemiology of Botrytis cinerea was studied in five annual strawberry crops using waiting-bed transplants, a system widely adopted in the Netherlands. On dead leaves of transplants the incidence of B. cinerea varied from 26.7% to 52.6%, but the leaf area with potential sporulation was low (3.5–15.6%). During each crop cycle, the availability of necrotic leaf substrate for spore production of B. cinerea was generally low and varied between seasons and with the quality of transplants. B. cinerea sporulated on a maximum of 15.5 cm² of leaf area per plant, measured as potential sporulation. The aerial concentration of B. cinerea conidia in untreated plots did not differ from the concentration in plots where all dead leaves had been removed, nor from the concentration at 25–50 m distance from the strawberry plots. B. cinerea incidence on flowers ranged from 5% to 96%, but no correlation was found with the potential spore production on necrotic leaves. Grey mould at harvest varied from 1.4% to 11.3% and was correlated with the average precipitation during the harvesting period but not with B. cinerea incidence on flowers. Post-harvest grey mould ranged from 2.1% to 32.6% and was correlated with petal colonisation by B. cinerea. The results suggest that in the annual cropping system with waiting-bed transplants, necrotic leaves are not a significant source of B. cinerea inoculum, unlike in other strawberry production systems. Therefore, control measures of grey mould in this annual system should focus on protection of flowers and young developing fruits, and not on the reduction of inoculum production on leaf debris.     

Wind dispersal of conidia of Botrytis spp. pathogenic to Vicia faba

Increasing numbers of conidia were blown from sporulating cultures of Botrytis fabae and B. cinerea as windspeed increased up to 10 m/s. Partial drying of cultures increased the number of spores blown away at low and intermediate windspeeds. Release of spores at a constant windspeed was sustained over a prolonged period. Different patterns of release from colonies of the two species when windspeed was gradually increased, or at a constant windspeed. may be related to differences in spore size affecting the drying rate. Many conidia of both species were released as clumps. A higher proportion of B. cinerea than B. fabae conidia were clumped, partly because the mean number of spores per clump was greater. Individual conidia fell more slowly in still air than did clumps. The humidity in a bean crop was more favourable to development of Botrytis lesions than that above the crop. Low windspeeds measured within crops may restrict dispersal of conidia and may result in uneven distribution of chocolate spot lesions.     

Effects of temperature,relative humidity and age of conidia on the incidence of spotting on gerbera flowers caused by Botrytis cinerea

In recent years, spotting of ray florets of gerbera flowers has become an important problem. This type of small necrotic lesions may occur before, but especially shortly after harvesting the flowers.Botrytis cinerea was easily isolated from such lesions. Inoculation withB. cinerea only gave typical necrotic lesions, when dry conidia were dusted on the flowers with a short period of high rh after inoculation. At 18–25 °C a high rh for at least 5 hours was necessary. Rotting of ray florets and receptacles byB. cinerea occurred when inoculated flowers were kept wet for a few days. Spots consist of one to several necrotic, usually epidermal cells. A single conidium could give rise to a necrotic lesion after germination. Germination of conidia and lesion formation occurred between 4 and 25 °C; at 30 °C, germination and lesion formation did not occur. Between 18 and 25 °C, many lesions became visible within 1 day after inoculation; at 4 °C it took 2 to 3 days before lesions could be seen. If kept dry, conidia ofB. cinerea remained ungerminated on ray florets of gerbera flowers and could be removed from the ray florets. Within 1 day at high rh, germination occurred and lesions were produced. Conidia ofB. cinerea, stored dry, were able to survive much longer than the lifetime of a gerbera flower. Even after storage at room temperature for up to 14 months, some conidia were able to germinate in vitro and on ray florets and induce the formation of lesions. Addition of gerbera pollen diffusate stimulated germination and lesion formation.     

Periodicity and gradients in dispersal ofAlternaria linicola in linseed crops

Conidia ofAlternaria linicola produced on infected linseed crops were mainly dispersed by wind. The numbers of conidia in the air above linseed crops collected by a Burkard spore sampler were greatest between 1200 h and 1300 h, when the relative humidity was lowest. Although numbers of conidia collected decreased with increasing height within and above the crop canopy, air-borneA. linicola conidia were present up to 80 cm above the crop canopy. Conidia ofA. linicola were transported by wind up to at least 40 m downwind from an artificial line inoculum source, but their numbers decreased with increasing distance from the source. In 1991, 1992, and 1993, the dispersal ofA. linicola conidia above linseed crops followed a seasonal periodicity which was influenced by weather conditions and cultural practices. The greatest numbers of conidia were collected during July, August and early September and coincided with periods favourable for sporulation and with an increase in the incidence of the disease in the senescent crop. Air-borneA. linicola conidia produced on point or line inoculum sources (naturally infected linseed stem debris) were responsible for the spread of the disease in linseed crops. In 1992 and 1993, the disease was first detected downwind from the sources, but by the end of the growing seasons, it had spread in all directions and up to 20 m and 60 m from the sources, respectively. Disease gradients were initially steep near the inoculum sources but they became flatter with time due to the secondary spread of the disease.     

Effect of chlorothalonil on resistance of Botrytis cinerea to dicarboximides in cucumber glasshouses1

Failures to control grey mould by dicarboximides on winter-grown cucumbers in glasshouses in Israel were attributed to the development of resistance to these fungicides in populations of Botrytis cinerea. A survey of 18 glasshouses in the winter of 1983-84 revealed that resistance is widespread where dicarboximides have been used extensively. During the summer, resistant B. cinerea populations often shift back to sensitivity; however, following few dicarboximide sprays in the subsequent winter, resistant strains reappear, increase rapidly and become dominant in the pathogen population. In two out of three glasshouse experiments, combination of chlorothalonil and dicarboximides in spray programmes slowed down the increase of resistant strains, whereas in a third experiment such a programme had no effect. The usefulness of chlorothalonil in fungicide-alternation programme to suppress dicarboximide resistance is dicussed in relation to the fitness properties of established versus newly-appearing resistant mutants.     

Resistance to Botrytis cinerea in Solanum lycopersicoides is Dominant in Hybrids with Tomato, and Involves induced Hyphal Death

Botrytis cinerea causes gray mold disease and affects hundreds of plant species, including tomato (Lycopersicon esculentum). The wild nightshade, Solanum lycopersicoides, is cross compatible with tomato and is more resistant to B. cinerea, thus representing a potential source for crop improvement. Tests involving droplet inoculation of detached leaves and spray inoculation of entire seedlings demonstrated that resistance to B. cinerea varies among S. lycopersicoides accessions, with S. lycopersicoides LA2951 being the most resistant accession tested. Expression of resistance in the intergeneric hybrid (L. esculentum cv. 'VF36' × S. lycopersicoides LA2951) suggested that resistance is at least partially dominant in tomato. A green fluorescent protein-tagged B. cinerea strain was used for confocal microscopic comparison of infection in leaves of S. lycopersicoides and tomato. Even though S. lycopersicoides supported spore germination, there was evidence for hyphal lysis and death 3 days after inoculation, at a time when lesions were expanding on susceptible tomato plants. The reduced frequency of B. cinerea lesion spread on S. lycopersicoides explains why this fungus produced fewer spores in this wild nightshade than in tomato.     

Influence of environmental factors on field concentrations of<Emphasis Type="Italic">Alternaria solani</Emphasis> conidia above a South African potato crop

Trends in weather variables and concentrations of airborne conidia ofAlternaria solani were monitored in a potato field in South Africa during three potato-growing seasons in 2001 and 2002. Distinct seasonal variation was noted, with a drop in spore numbers during winter. Peaks in spore concentration coincided with periods favorable for spore formation and dispersal; most notable was the effect of interrupted wetting periods. Diurnal periodicity of spore dispersal was also observed, with the peak of spore concentrations between 9h00 and 18h00. Few spores were sampled at night, when wind velocity and temperature are lowest and relative humidity is highest. Increased numbers of spores were sampled during days of harvesting or when other ground-operated farm equipment was used. The results obtained in this study will be useful in establishing decision support systems to control early blight on potatoes in southern Africa. http://www.phytoparasitica.org posting July 10, 2003.     

The Effect of Treatment With Ulocladium Atrum on Botrytis Cinerea-attack of Geranium (Pelargonium Zonale) Stock Plants and Cuttings

In two successive seasons, the effect of treatment of geranium stock plants with the competitive saprophytic fungus Ulocladium atrum as a biocontrol agent against Botrytis cinerea was compared to a fungicide treatment with Euparene M. B. cinerea incidence and severity on the stock plants, B. cinerea spore load in the air around stock plants and death of cuttings due to B. cinerea were scored. B. cinerea incidence and severity were much stronger in the second than the first experiment. This was quantitatively expressed by higher numbers of conidia of B. cinerea monitored in the second than the first year, both on necrotic (a maximum for the control of 27.5 × 10⁶ spores per sample - all necrotic leaves of five plants - in experiment 1 against 86 × 10⁶ in experiment 2) and green leaves, but numbers of conidia of B. cinerea recovered from the air were only slightly different. The death rate of cuttings was moderate in the first and extremely high in the second experiment. For the fungicide treatment, maximum sample values of 7% and 76% of 6-week old cuttings were killed in the first and the second experiment respectively. Treatment with U. atrum was effective in reducing all parameters studied. With the exception of the spore load of B. cinerea in the air and the success of cuttings, the effect of U. atrum varied from as good as the fungicide to half as effective. In the first trial, only Euparene M reduced spore load in the air, in the second trial only U. atrum consistently did so. In the first trial U. atrum reduced death of 4-week old cuttings, though less than fungicide (1.2, 20 and 38% killed with fungicide treatment, U. atrum treatment and control respectively). In the second trial only the fungicide reduced loss of cuttings. The impact of the data on the integration of U. atrum in a control system of B. cinerea in geranium is discussed.     

Preventive and post‐infection control of Botrytis cinerea in tomato plants by hexanoic acid

The antifungal activity of hexanoic acid on the phytopathogen Botrytis cinerea was studied. This chemical inhibited both spore germination and mycelial growth in vitro in a concentration‐ and pH‐dependent manner, and stopped spore germination at a very early stage, preventing germ‐tube development. The minimum fungicidal concentration (MFC) for in vitro spore germination was 16 mm . Hexanoic acid also inhibited in vitro mycelial growth of germinated spores at an MFC of 12 mm . Studies performed to characterize the mechanisms underlying the antimicrobial effect of hexanoic acid showed that it alters fungal membrane permeability. In addition, hexanoic acid treatment increased the levels of spermine, spermidine, putrescine and cadaverine in B. cinerea mycelia. Spray application of hexanoic acid at fungicidal concentrations on 4‐week‐old tomato plants prior to fungal inoculation reduced necrosis diameter by approximately 60%. Application of the same hexanoic acid concentrations on previously infected plants reduced further necrosis expansion by around 30%. The results suggest that this chemical acts as a preventive and curative fungicide. Interestingly, treatments with hexanoic acid at concentrations below the MFC in hydroponic solution prior to fungal inoculation significantly reduced necrosis area. These results suggest an inducer effect of plant responses for hexanoic acid treatments at these concentrations. Hexanoic acid is a good candidate for safe antifungal treatments for the control of B. cinerea, which is responsible for many economic losses on fruits, vegetables and flowers.     

Suppression of potato cyst nematode root penetration by the endoparasitic nematophagous fungusHirsutella rhossiliensis

The endoparasitic nematophagous fungusHirsutella rhossiliensis was tested for its ability to suppress root penetration and cyst formation by the potato cyst nematode speciesGlobodera pallida. Isolates ofH. rhossiliensis were obtained from infected potato cyst nematode juveniles from different starch potato fields in The Netherlands. The isolates showed no difference in spore adhesion to juveniles on agar plates (adhesion rate: ±90%). The most rapid growing isolate, CBS 108.94, was used for experiments. Vegetative mycelial colonies ofH. rhossiliensis CBS 108.94, grown in potato dextrose broth, were used as soil inoculum. During submerged cultivation the mycelial colonies produced phialides (spore-bearing cells) but no spores. Exposed to the air, however, spores were rapidly formed. The effect of different soil inoculum densities of mycelial colonies on root penetration byGlobodera pallida was examined in an experiment in 250-ml pots. Up to a mycelial colony concentration representing a potential spore density of 10⁴ g^–1 soil no suppression occurred. At approximated densities of 2.5×10⁴ and 10⁵ spores g^–1 soil the numbers of juveniles which penetrated roots were reduced by 30% and 34%, respectively. The distribution of the inoculum could be improved by fragmentation of the mycelial colonies before soil inoculation. Using mycelial fragments, again no suppression of root penetration was observed up to a potential spore density of 10⁴ g^–1 soil, but at densities of 10⁵ and 10⁶ g^–1 a suppression of 54% and 88%, respectively, was measured. In a greenhouse experiment, soil inoculation with mycelial colonies with a potential spore production of 2.5×10⁵ g^–1 soil resulted in a suppression of root penetration of 37% and 51% after 5 and 6 weeks, respectively, but the number of newly formed cysts after 18 weeks in soil was not different for control and inoculated pots. It is concluded thatH. rhossiliensis may be useful for the reduction of root damage caused by juveniles of potato cyst nematodes, but the usefulness for population control is doubtful.     

Studies on red light-induced resistance of broad bean to Botrytis cinerea: I. Possible production of suppressor and elicitor by germinating spores of pathogen

When detached broad bean leaves were preinoculated with virulent strain B304 of Botrytis cinerea 24 h before a challenge inoculation with strain B304, lesion formation by B304 was significantly inhibited in red light but not in the dark. In leaves that were preinoculated with avirulent strain 021 and then challenged by B304, however, lesion formation was not inhibited even under red light. Such differences in lesion formation after the challenge inoculation with an avirulent strain were also observed with lesions caused by Alternaria alternata, a nonpathogen of broad bean and by avirulent strain 021R in the presence of germination fluid from spores of strains B304 and 021R. These results suggest the possibility that virulent B. cinerea produced a suppressor involved in induced susceptibility and an elicitor involved in resistance induced by red light during spore germination.