Microbial population dynamics on Golden Delicious apples from bud to harvest and effect of fungicide applications

The microbial population dynamics on apples cv. Golden Delicious were analysed every 15 days between bud and harvest in a fully replicated experiment in northern Spain in 1994 and 1995. The total microbial populations varied with developmental stage, and with prevailing climatic conditions. The predominant mycroflora were the filamentous fungi Cladosporium and Alternaria spp. and white and pink yeasts. Other genera isolated included mainly species of Epicoccum, Fusarium and Acremonium. However, the most important post-harvest pathogens Penicillium expansum and Botrytis cinerea were seldom isolated from ripening apples. Maximum total filamentous fungal populations occurred after fruit set and during early ripening [2 × 10⁴cfu (colony-forming units) g^-1 approximately] while those of bacteria were maximum at bud stage (3.5 × 10⁵and 3.0 × 10⁴ cfu g^-1 in 1994 and 1995 respectively). White yeasts were more numerous than pink yeasts. Endophytic infection of apple buds by Alternaria spp., responsible for core rot, was found in almost all bud tissue. By contrast, Cladosporium spp. were initially isolated later from 12.5–50% of tissue samples during blooming and fruit set. The impact of a four-spray fungicide regime during apple development significantly decreased the total filamentous fungal populations in both years, and that of Cladosporium spp. in 1994. However, bacterial populations were often higher on apples from fungicide-treated plots. Fungicide sprays decreased populations of Cladosporium, Alternaria and white yeasts for a maximum of up to 15–30 days after application. Fungicide application had little effect on endophytic infection of apples by Alternaria spp. between bud and harvest.     

The effect of a post-harvest heat-treatment on the storage behaviour of Cox's Orange Pippin apple fruits

Samples from a wide range of commercially grown Cox's Orange Pippin apple fruits were immersed in water at 45° C. for 10 min. to control rotting due to Gloeosporium spp. In two seasons the incidence of rotting following storage for 19 weeks at 3·3° C. (38° F.) was reduced to about 40% of the level in the untreated samples. However, there was a marked increase in the incidence of breakdown and core flush in the treated fruit, particularly when stored in a controlled atmosphere of 5% CO₂, 3 % O₂. A similar but less severe increase in the incidence of breakdown was also recorded in the second season in treated fruit stored in 2% O₂ (with continuous removal of CO₂). The treated fruit tended to develop a more yellow ground colour and softened more rapidly during storage in 5% CO₂, 3% O₂. It is concluded that the treatment advances fruit senescence and that the increased incidence of physiological disorders is likely to limit the use of this method for the control of Gloeosporium rotting in English-grown Cox apples.     

Treatment with chlorous acid to inhibit spores of Alicyclobacillus acidoterrestris in aqueous suspension and on apples

Aim: To test the efficacy of a chemical (chlorous acid) for reducing the numbers of viable Alicyclobacillus acidoterrestris spores in laboratory media and on apples. Methods and Results: Alicyclobacillus acidoterrestris spores in aqueous suspension and on apple surfaces of four different cultivars were treated with 268 ppm chlorous acid. Treatment with 268 ppm chlorous acid sharply reduced the numbers of spores of A. acidoterrestris in laboratory media by 1·6, 4·3, and 7·0 log₁₀ reductions for 5, 10, and 15 min treatments, respectively. Chlorous acid also effectively reduced the spore load on apple surfaces. Alicyclobacillus acidoterrestris spore counts were significantly reduced by about 5 log₁₀ after 10 min treatment on four different apple cultivars (‘Red Delicious’, ‘Golden Delicious’,’ Gala’, and ‘Fuji’). There was no synergistic effect on spore reduction when chlorous acid treatment was combined with heat. Conclusions: These results show that chlorous acid is highly efficacious against A. acidoterrestris spores on apple surfaces. Significance and Impact of the Study: Chlorous acid can be used as an alternative sanitizer of chlorine to control a major A. acidoterrestris contamination source in juice processing plants.     

Mango stem end rot pathogens — Infection levels between flowering and harvest

During flowering and fruit set of mango (Mangifera indica L.), colonisation by fungi (Alternaria alternata, Cladosporium cladosporioides, Dothiorella dominicana, Dothiorella mangiferae, Dothiorella sp., Epicoccum purpurascens and Pestalotiopsis sp.) increased as the flowers senesced and young fruit formed. In the third week after flowering, the incidence of Dothiorella dominicana and Dothiorella mangiferae associated with mango fruit-pedicel connection tissue declined coincidentally with early fruit-fall, suggesting that early infections by Dothiorella spp. may cause fruitlet abortion. Dothiorella spp. levels in fruit-pedicel connection tissue remained low for the subsequent 6 weeks, after which they increased. By 16 weeks after flowering, the incidence of Dothiorella spp., determined by isolation from fruit-pedicel connection tissue, was similar to the incidence of stem end rot caused by Dothiorella spp., which developed in fruit harvested at that time. By contrast, the frequency of detection of Dothiorella spp. in peduncle tissue peaked 11 weeks after flowering, when the levels of stem end rot developing in fruit were already similar to the levels recorded in fruit harvested at 16 weeks and later. The results suggest that in fruit and fruit-pedicel tissue, colonisation might arise from Dothiorella spp. occurring endophytically in the peduncle. The earliest indicator of stem end rot incidence at harvest was the infection level in peduncle tissue sampled 11 weeks after flowering. Early assays of peduncle tissue for Dothiorella spp. might prove useful for selecting crops with low stem end rot infection levels.     

Control of Postharvest Diseases of Sweet Cherry with Ethanol and Hot Water

Complete inhibition of the germination of spores of Penicillium expansum occurred after 10 s exposure to 40% ethanol or more at ambient temperature, while spores of Botrytis cinerea were completely inhibited by 30% ethanol or more. Mortality of the spores of P. expansum and B. cinerea in heated 10% ethanol was higher than in water at the same temperatures. Immersion of naturally inoculated fruit in 20, 30, 40, or 50% ethanol reduced the decay present after storage for 10 days at 20°C similarly and by approximately 60–85%. Immersion of fruit that had been inoculated with the spores of P. expansum and B. cinerea reduced decay by both pathogens after storage for 30 days at 0°C and 5 days at 20°C when 30% or higher concentrations of ethanol were used. The incidence of decay after immersion in water alone for 30 s at 24, 50, 55, or 60°C was 57.7, 44.7, 46.2, and 35.7%, respectively, while 10% ethanol at these temperatures the decay incidence to 52.2, 33.9, 32.8, or 14.7%, respectively. Water treatments at 50, 55, or 60°C alone were not effective against P. expansum, while their efficacies were significantly increased by the addition of 10% ethanol. The most effective treatment was immersion in 10% ethanol at 60°C. Ethanol treatments at 20, 30, 40, or 50% and water treatments at 55 or 60°C significantly reduced natural fungal populations on the surfaces of fruit in all of the experiments. Addition of 10% ethanol to water significantly increased the efficacy of water in reducing the fungal populations at elevated temperatures. None of these treatments caused surface injuries to the fruit or adversely affected stem colour.     

Isolation and pathogenicity of some South African entomopathogenic fungi (Ascomycota) against eggs and larvae of Cydia pomonella (Lepidoptera: Tortricidae)

Virulent entomopathogenic fungal strains against Cydia pomonella (Lepidoptera: Tortricidae) were isolated and identified in the Western Cape Province of South Africa. Thirty-nine isolates belonging to six species were obtained using the insect bait method. Generally, Metarhizium robertsii (Ascomycota: Hypocreales) was the most frequently encountered species representing 51% of the total number of isolates collected from the soil samples. This is the first report of M. robertsii from southern Africa. Mortality data from an immersion bioassay indicated that the 39 fungal isolates were pathogenic against fifth instar larvae of C. pomonella inducing 47–85% insect mortalities. Two M. robertsii strains, MTL151 and GW461, induced 85% larval mortality and were selected for further evaluations. The exposure of freshly laid eggs to wax papers that were pre-treated with fungal spores ranging from 10³ to 10⁸ spores ml^?1 of MTL151 and GW461 resulted in a significant reduction of egg hatchabilities, from 95 to 66% and 93 to 71%, respectively as spore concentration increased. First instar larval neonates were exposed to apples that were pre-sprayed topically with varied conidia suspensions (10³?10⁸ spores ml^?1). The mean percentage of participating apples with larvae in their cores/flesh significantly reduced from 53 to 10% (MTL151) and 76 to 10% (GW461) of 10 apples, and a concurrent decrease in the incidence of apple fruit rot was observed as conidia concentration increased. Up to 90% of apples treated with 1 × 10⁸ spores ml^?1 had no larvae present in their cores and this result compared favourably with the commercial pesticide Fruitfly®, containing the active ingredient cypermethrin (20 g/l) used at a recommended dose of 0.25 ml/250 ml of water.     

Identification of two Bacillus amyloliquefaciens strains with high suppression to the key fruit pathogens of Chinese jujube

Two bacterial strains (ZJ01 and ZJ02) isolated from the phyllosphere of Chinese jujube were tested for antifungal activities and morphological mechanisms against Phoma destructiva, Alternaria alternata and Fusicoccum spp., three key pathogens of jujube fruits. Based on their physiological characteristics and 16S rDNA gene sequences, both strains were identified as Bacillus amyloliquefaciens. The disease incidence of jujube fruits sprayed with the fermentation liquor of ZJ01 and ZJ02 on the puncture wounds before inoculation with the spores of those three key pathogenic fungi was 10% or 66.7%, significantly lower than that of Nutrient Broth medium treatment (100%). These results suggest that strains ZJ01 may be a potential biological control agent for Chinese jujube fruit diseases. The antagonistic bacteria could cause five kinds of hyphal alterations of the causal agents, the maximum number ever observed. Of them, hyphal swelling and excessive branching were the dominant phenomenon, the other three varied with the pathogens. The diverse morphological malformation might be helpful to understand the broad antifungal activity of Bacillus spp..     

Effect of orchard and postharvest application of daminozide on ethylene synthesis by apple fruit

The effects of daminozide (butanedioic acid-2,2-dimethylhydrazide) on ethylene synthesis by apple fruits were investigated. The objective was to determine the effects of postharvest applications as compared to the standard application of diaminozide in the orchard. Immersion in a solution containing 4.25 g L^?1 active ingredient for 5 min delayed the rise in ethylene production in individual “Cox” apples at 15°C by about 2 days, whereas orchard application of 0.85 g L^?1 caused delays of about 3 days. Both modes of application depressed the maximal rate of ethylene production attained by ripe apples by about 30%. Daminozide did not affect the stimulation of respiration by ethylene treatment of “Gloster” apples, but it delayed the increase in ethylene synthesis. Daminozide applied immediately after harvest delayed the rise in ethylene synthesis in “Golden Delicious” held at 15°C, but it was less effective when applied 48 h after harvest or when apples were held at 5°C. Exposure to 1–2 μl L^?1 ethylene for 48 h was less effective in promoting the rise in ethylene in daminozidetreated “Cox” and “Gloster” apples than in untreated fruit. High (100–1000 μl L^?1) concentrations of ethylene more or less overcame the daminozide effect. Apples absorbed about 40% of surface-applied [¹⁴C]daminozide in 48 h, but more than 90% of the radioactivity in the fruit was recovered from the peel and outer 1 cm of the cortex. Daminozide was partly converted to carbon dioxide and other metabolites.     

Vesicular-arbuscular mycorrhizal spore populations in sugar maple (Acer saccharum marsh. L.) forests

 The numbers and types of spores of vesicular-arbuscular mycorrhizal fungi occurring in the top 15 cm of the soil in three maple forests in Eastern Canada were investigated using traditional wet-sieving/decanting methods. In the most acid site, at St. Hippolyte, Québec, where the soil had been amended with base cations, after 1 year there was no effect on the numbers of spores present. Vesicular-arbuscular mycorrhizal spores present at St. Hippolyte consisted of Glomus rubiforme, other Glomus spp. and Acaulospora spp. Although the sporocarpic species, G. aggregatum, G. macrocarpum and G. rubiforme occurred at St. Hippolyte, they were not found at the two less acid sites (Waterloo, Ontario and Lacolle, Québec) . Spores of Acaulospora spp. were found at all three sites, but were most abundant at St. Hippolyte. At St. Hippolyte the total number of spores was much higher than at the other two sites; at Waterloo numbers were an order of magnitude lower than at St. Hippolyte. It is suggested that G. rubiforme and Acaulospora species may be adapted to acid conditions. Seasonal patterns of spore abundance suggested that Acaulospora spp. may sporulate during the spring, whereas G. rubiforme may sporulate during the fall. Accepted: 6 September 1995     

Life from the ashes: survival of dry bacterial spores after very high temperature exposure

We found that spores of Bacillus amyloliquefaciens rank amongst the most resistant to high temperatures with a maximum dry heat tolerance determined at 420 °C. We found that this extreme heat resistance was also maintained after several generations suggesting that the DNA was able to replicate after exposure to these temperatures. Nonetheless, amplifying the bacterial DNA using BOXA1R and (GTG)₅ primers was unsuccessful immediately after extreme heating, but was successful after incubation of the heated then cooled spores. Moreover, enzymes such as amylases and proteases were active directly after heating and spore regeneration, indicating that DNA coding for these enzymes were not degraded at these temperatures. Our results suggest that extensive DNA damage may occur in spores of B. amyloliquefaciens directly after an extreme heat shock. However, the successful germination of spores after inoculation and incubation indicates that these spores could have a very effective DNA repair mechanism, most likely protein-based, able to function after exposure to temperatures up to 420 °C. Therefore, we propose that B. amyloliquefaciens is one of the most heat resistant life forms known to science and can be used as a model organism for studying heat resistance and DNA repair. Furthermore, the extremely high temperature resistivity of these spores has exceptional consequences for general methodology, such as the use of dry heat sterilization and, therefore, virtually all studies in the broad area of high temperature biology.

     

Effects of Treatment with a Composite Preparation (2-Chloroethylphosphonic Acid and Methacide) or Butylated Hydroxyanisole on Ethylene Release in Apples

We studied the effect of a Russian composite preparation (2-chloroethylphosphonic acid and methacide) and butylated hydroxyanisole on ethylene release in whole fruit and peel disks of two apple cultivars, Antonovka obyknovennaya (Antonovka) and Simirenko's rennet (Simirenko). Treatment with the composite preparation was followed by an increase in ethylene release from both the whole apples and peel disks. The development of microbial infection (fruit rot) in the whole apples became less pronounced after the treatment. Treatment of whole apples with the antioxidant butylated hydroxyanisole (BHA) increased the intensity of ethylene release during the first subsequent days; thereafter, ethylene release decreased and was 10–15% lower than in the control on days 10–12. In model experiments, BHA decreased ethylene release from apple peel disks below control levels as early as on the first day after treatment. Antonovka apples gave quick responses to the treatment. In the late-ripening Simirenko apples, the response persisted for a longer period. Our results suggest that treatment with physiologically active preparations affects the ethylene release, ripening, and preservation of apples in storage.     

Observations on the infection and rotting of apples var. Bramley's Seedliong by Diaporthe perniciosa

In N. Ireland Diaporthe perniciosa March is a common inhabitant of the dead wood of Bramley's Seedling apple trees, and its pycnospores, extruded throughout the growing season, are responsible for fruit infections which result in stalk-end rots during storage.
Pycnidia were produced in culture only after exposure to light, near-UV radiation being particularly favourable. The carbon source in the medium influenced pycnidia production: high yields were obtained where mannitol was used. Only a -spores germinated under the conditions tested, giving optimum germination at 25 C. in a water film. No germination occurred at less than 98% R.H. Germ tubes and hyphae developed appressoria after germination on artificial surfaces but on apple skin most spores produced only a protuberance from which penetration hyphae developed directly. The cuticle and epidermal cells of apples of all ages were penetrated within a few days of inoculation. Intact surfaces and lenticels were penetrated equally. Susceptibility to further rotting increased with age of the fruit. Apples loaded with spores early in the season rotted later than similar apples loaded just before picking. Polygalacturonase was detected in sap expressed from rotted apples of various ages, the level of activity being correlated with the susceptibility of the fruit.     

Prior experience affects the visual ability ofRhagoletis pomonella flies (Diptera: Tephritidae) to find host fruit

Females of the apple maggot fly,Rhagoletis pomonella, were allowed for 3 days to alight upon and oviposit in green or red 18- to 20-mm hawthorn host fruit (Crategus mollis) or green or red 45- to 55-mm apple host fruit (Malus pumila) hung from branches of potted host trees in field enclosures. Subsequently, when females were released individually on potted host trees harboring fruit of one of these types, their ability to find fruit of unfamiliar size proved unaffected by prior experience with fruit but their ability to find fruit of unfamiliar color was significantly affected. Specifically, females exposed to red hawthorns or red apples were less able to find green hawthorns or green apples than were females experienced with either of the latter fruit types. Fruit odor was found to have no effect on female ability to find familiar compared with unfamiliar green fruit. In contrast, a difference in size (or surface chemistry) between familiar and unfamiliar fruit but not a difference in fruit color had a significant negative influence on the propensity of alighting females to bore into unfamiliar fruit. Three bouts of experience with alighting upon and ovipositing into fruit over a period of about 1 h had no detectable effect on female ability to find unfamiliar fruit but did reduce propensity to bore into unfamiliar fruit. Our findings are discussed in relation to insect ability to learn visual and chemical stimuli of resources and insect propensity to form host races. We also discuss the potential impact of our findings on nonpesticidal, behavioral methods of managingR. pomonella in commercial apple orchards.     

Influence of pectinase treatment on fruit spirits from apple mash, juice and pomace

The current investigation was conducted to determine the influence of pectinase treatment on fruit spirits produced from apple mash, juice, and pomace. Crispin apples were processed into apple mash, juice, and pomace in our pilot-plant, and fermented with a commercial Red Star wine yeast (Sachharomyces cerevisiae Davis 904). After fermentation, the samples of fermented apple mash, juice, and pomace were distilled, and the distillates were analyzed by HPLC with a Bio-Rad Aminex HPX 87H column and a refractive index detector. Methanol, ethanol, n-propanol, iso-butanol, and iso-amyl alcohol were identified as the major alcohols in all the apple spirits. Student's t-test results indicate that there are significant differences between the methanol concentrations of pectinase treated and non-pectinase treated apple spirits. Duncan's multiple range tests show significant differences in the concentrations of methanol of the fruit spirits made from apple mash, juice, and pomace. Apple pomace yielded significantly higher methanol concentrations than apple mash and juice. Pectinase treatment had little effect on the concentrations of n-propanol, iso-butanol, and iso-amyl alcohol. It is concluded that fruit spirits made from the pectinase treated mash, juice, and pomace of Crispin apples had methanol concentrations significantly above the United States FDA guidance of 0.35% by volume or 280 mg/100 mL of fruit brandy containing 40% ethanol.     

An assessment of some new treatments for the control of rotting of stored apples

Benomyl (0.025% a.i.) and thiabendazole (0.05 % a.i.) applied in July, August and September to apple trees cvs. Sunset and Cox's Orange Pippin gave slightly better control of Gloeosporium spp. rots in stored fruit than captan (0.1% a.i.) applied at similar intervals. On Cox's Orange Pippin, treatment with captan at 0.1 % a.i. in July and at 0.2% a.i. in August was as effective as three sprays at 0.1 % a.i. in July, August and September in controlling both Gloeosporium spp. and Nectria galligena rots. Benomyl applied in the spring and early summer to control apple scab on orchard trees reduced the incidence of Gloeosporium spp. and Monilia fructigena in stored fruit. Pre-harvest sprays of benomyl and captan gave good control of rotting caused by N. galligena but thiabendazole was ineffective. Post-harvest dips of thiabendazole controlled rotting by Gloeosporium spp. butnot byN. galligena.     

Dacus dorsalis flies can learn to find and accept host fruit

Mature oriental fruit fly females, Dacus dorsalis, from a population cultured on host fruit in the laboratory for one generation responded positively to visual and olfactory stimuli of individual natural kumquat (Fortunella japonica) and apple (Malus pumila) host fruit or models of these fruit hung from branches of potted trees in field enclosures. When females were exposed for 3 days to natural kumquats or apples on trees and subsequently released individually onto trees harboring one or the other of these fruit types, a significantly greater proportion of those exposed to kumquats than those exposed to apples or those not exposed to any fruit visited kumquats. Females exposed to kimquats for at least 3 days followed by at least 3 days of exposure to apples retained ability to find kumquats. Compared with females exposed to apples for 3 days or with naive females, females exposed to kumquats for 3 days exhibitied no less ability to find apples but did significantly refrain from accepting apples for oviposition. A final test was conducted in which females were exposed to natural kumquats or apples for 3 days and tested for response to inanimate models of either the same color and size as natural kumquats (orange, 20-mm diameter) or apples (green, 75-mm diameter) or models of the same color but opposite size. Results suggest that fruit size is the principal character learned and used in finding kumquats, which apparently are somewhat inconspicuous to an inexperienced foraging D. dorsalisfemale.     

Effects of treatment with a composite preparation (2-chloroethylphosphonic acid and methacide) or butylated hydroxyanisole on ethylene release in apples

We studied the effect of a Russian composite preparation (2-chloroethylphosphonic acid and methacide) and butylated hydroxyanisole on ethylene release in whole fruit and peel disks of two apple cultivars, Antonovka obyknovennaya (Antonovka) and Simirenko's rennet (Simirenko). Treatment with the composite preparation was followed by an increase in ethylene release from whole apples and peel disks. The development of microbial infection (fruit rot) in whole apples became less pronounced after the treatment. Treatment of whole apples with the antioxidant butylated hydroxyanisole (BHA) increased the intensity of ethylene release during the first subsequent days; thereafter, ethylene release decreased and was 10-15% lower than in the control on days 10-12. In model experiments, BHA decreased ethylene release from apple peel disks below control levels as early as on day 1 after the treatment. Antonovka apples gave quick responses to the treatment. In the late-ripening Simirenko apples, the response persisted for a longer period. Our results suggest that treatment with physiologically active preparations affects ethylene release, ripening, and preservation of apples in storage.     

Control of green mould of citrus by using Trichoderma harzianum,humic acid or garlic

Penicillium digitatum, an aggressive fungus causes post-harvest decay of mandarin sweet orange and Washington navel. In vitro Trichoderma harzianum or humic acid (HA) or powdered cloves of garlic caused inhibition of fungal growth of isolates P₁ and P₂. Under storage conditions, the fruit citrus is protected by using T. harzianum with standard volume 2.0?ml (9.6?×?10⁶?conidia/ml) and application 24?h before inoculation reduces disease incidence and disease severity after seven?days from inoculation with P. digitatum spore suspension (1.0?×?10⁶?spores/ml) compared to control. Spraying the fruit citrus by standard volume of 2.0?ml of either HA or powder cloves of garlic 1% on each fruit 24?h before inoculation reduces disease incidence and disease severity after seven?days from inoculation with P. digitatum (1.0?×?10⁶ spores/ml) compared to control. The lowest percentage of disease incidence and disease severity were associated with powder of cloves garlic and followed by HA and T. harzianum during two growing seasons compared with the untreated and control.     

The effect of store conditions on the rotting of apples, cv. Bramley's Seedling, by Nectria galligena

The log of the time interval between inoculation with Nectria galligena in October and the onset of rotting in apples held in air was proportional to the deficit between the temperature of incubation and 25°C, but temperature did not affect the rate of subsequent rot expansion. Rots expanded equally fast whether apples were held in dry or moist air. The quantity of rotted tissue obtained after incubating inoculated apples in atmospheres containing up to 12.5% CO₂ increased with increasing concentrations of CO₂ greater than 2.5%. The quantity of rotted tissue obtained in apples incubated in 10% CO₂ was three times as great as that obtained after incubation in air. The incidence of natural rots was lower in apples stored at 4% CO₂ than in those stored in air and rotting increased with increasing concentrations of CO₂ higher than 4%. Colonies of N. galligena grew faster on malt agar plates incubated in 5% CO₂ than in air, but growth was slower in 10% CO₂ than in air. The quantity of benzoic acid per mg hyphae accumulated in developing lesions was similarly related to the CO₂ concentrations up to 2.5% but decreased at higher concentrations, and the quantities found in apples stored in CO₂ concentrations >5.0% CO₂ were less than in those stored in air.     

The role of wounds in the infection of oranges by Penicillium digitatum Sacc

Inocula of spores of Penicillium digitatum in water applied to apparently uninjured skin of oranges do not cause lesions to develop. Addition of citric acid, orange juice, or various extracts of rind had little effect on susceptibility to infection. When spores in water are applied to wounds made between oil vesicles, lesions develop only from wounds that penetrate deeply into the albedo. The flavedo of most oranges seems to be resistant to infection even when damaged, but in a few consignments it showed much less resistance. Increasing the number of conidia in the inoculum caused more lesions to develop but some fruits developed lesions from inocula containing very few spores. The method and timing of spore application to wounds had a considerable effect on the incidence of lesions; emanations from infected fruit had no effect. Lesions developed more rapidly and readily when suspensions of spores in water were applied to wounds in the skin that damaged oil vesicles; wounds as shallow as 0–25 mm allowed lesions to develop.