Application of <Emphasis Type="Italic">Pantoea agglomerans</Emphasis> CPA-2 in combination with heated sodium bicarbonate solutions to control the major postharvest diseases affecting citrus fruit at several mediterranean locations

We determined the potential of using a formulated product based on Pantoea agglomerans CPA-2, either alone or in combination with heated sodium bicarbonate (SBC) solutions, to control the major postharvest diseases affecting citrus crops in the Mediterranean region. Treatments applied either individually or in combination were tested in semi-commercial and commercial trials carried out with oranges and mandarins from the Algarve, Andalusia and Catalonia. Firstly, several formulations of the biocontrol agents were tested in laboratory trials; one of them, a freeze-dried formulation of P. agglomerans strain CPA-2 called FD10-3, was chosen for combined with SBC. This formulation, applied at 2 × 10⁸cfu ml⁻¹ and the SBC treatment, applied at 3% 50°C for 20–40 s, demonstrated that it was possible to reduce decay development in laboratory trials. Semi-commercial applications of FD10-3 and 3% SBC solution at 50°C for 40 s showed excellent control of decay in unwounded mandarins and oranges artificially inoculated with both Penicillium digitatum and P. italicum. No rind injuries or residues attributable to hot water or SBC were observed on treated fruits. Combined treatment provided better control than the two treatments applied separately. Commercial trials demonstrated an important reduction in natural decay with the treatment of SBC 3% at 50°C for 40 s. Furthermore, bacterial-product formulation treatment significantly reduced the percentage of infected fruit and in some cases this reduction was equal to chemical treatments. Even so, no improvement in efficacy was observed with the combination of FD10-3 and SBC in the commercial test. We also assessed the ability of FD10-3 to grow at the wound site in oranges, whether alone or in the presence of SBC, and also its compatibility with standard citrus packinghouse practices.     

Modes of Action of Pantoea agglomerans CPA-2, an Antagonist of Postharvest Pathogens on Fruits

Pantoea agglomerans CPA-2 is an effective antagonist against the postharvest pathogens Penicillium digitatum and Penicillium italicum on citrus fruits but its mode of action is unknown. Possible mechanisms studied in this work were antibiosis, induced resistance, competition and production of chitinolytic enzymes. P. agglomerans CPA-2 was unable to produce antibiotics or chitinolytic enzymes under the conditions tested. Induction of resistance by P. agglomerans CPA-2 was studied in oranges by measuring phenylalanine ammonia lyase and peroxidase enzyme activity in the orange peel at different time points after inoculation with the antagonist and/or the pathogen. No significant augmentation of enzyme activity after inoculation of oranges with P. agglomerans CPA-2 in the presence or absence of the pathogen was observed. P. agglomerans was effective only when it is in close contact with the pathogens. Competition for nutrients was studied using tissue culture plates with cylinder inserts, which allowed competition for nutrients to be studied without competition for space since physical contact between pathogen and antagonist was avoided. The presence of P. agglomerans in the tissue culture wells clearly decreased the germination of Penicillium conidia present in the cylinder when diluted orange peel extract or diluted potato dextrose broth was the nutrient source. Germination of Penicillium conidia, however, was almost completely inhibited when pathogen and antagonist were in physical contact. These results indicate that competition for nutrients is one of the modes of action of P. agglomerans CPA-2, but that physical contact between pathogen and antagonist is important for effective control.     

The use of sodium carbonate to improve curing treatments against green and blue moulds on citrus fruits

The effectiveness of curing oranges and lemons at 33 degrees C for 65h followed by storage under ambient and cold-storage conditions was investigated. This treatment effectively reduced the incidence of Penicillium digitatum (Pers) Sacc and P italicum Wehmer decay on inoculated and naturally infected oranges and lemons stored at 20 degrees C for 7 days. However, it failed to control green and blue mould infections on fruits placed in long-term cold storage, except green mould on oranges, which was effectively controlled. Dipping fruits in a sodium carbonate solution (20 g litre(-1)) for 2.5 min following a curing treatment at 33 degrees C for 65 h satisfactorily reduced green and blue mould incidence during subsequent long-term storage at 4 degrees C on oranges and at 10 degrees C on lemons. The efficacy was greater on injured fruits inoculated after the combination of treatments was applied, achieving a 60-80% reduction in decay in comparison with the curing treatment alone in all cases. A significant reduction of blue mould was also observed on fruits inoculated both before the treatments and on those re-inoculated after the treatments, demonstrating both protectant and eradicant activity. Thus, combining curing at 33 degrees C for 65 h with sodium carbonate treatment effectively controlled these post-harvest diseases on artificially inoculated citrus fruits and protected against re-infection. With naturally inoculated lemons, curing followed by sodium carbonate significantly reduced both green and blue mould incidence, but was not superior to curing alone. With naturally infected oranges, curing significantly reduced blue mould, but decay was not reduced further when followed by sodium carbonate treatment.     

21种药用植物提取物对柑橘青霉病抑菌作用的筛选及白薇醇提物对脐橙青霉病的防治效果

为开发研制新型植物源柑橘保鲜剂,以抑菌圈直径为指标,利用牛津杯法筛选出对柑橘采后意大利青霉有良好抑制作用的21种中草药的醇提物,并对白薇醇提物防治脐橙青霉病的效果进行了评价。结果表明:白薇、零陵香、鸦胆子、藏青果、白药子5种中草药乙醇提取物对意大利青霉均有较好的抑制效果,抑制圈直径达26 mm以上,其中白薇醇提物抑菌活性最佳,抑制圈直径可达38.39 mm,最小抑菌浓度和最小杀菌浓度分别为3.13 mg/m L和6.25 mg/m L,且抑菌效果与对照药剂1 000倍咪鲜胺无显著差异。25 mg/m L白薇醇提物处理脐橙7 d后,对脐橙青霉病的防治效果达35.93%,与1 000倍咪鲜胺的防治效果相当(36.37%);白薇醇提物处理后可抑制意大利青霉菌菌丝生长,导致菌丝肿胀、膨大、畸形、细胞膜透性增加、细胞内含物外渗;还可诱导脐橙体内过氧化物酶活性下降,而苯丙氨酸解氨酶和几丁质酶活性上升。表明白薇醇提物对脐橙采后青霉病具有较好的防治效果,可进一步开发为植物源柑橘保鲜剂。     

RETRACTED ARTICLE: Effect of calcium chloride on the biocontrol efficacy of two antagonistic yeasts against <Emphasis Type="Italic">Penicillium expansum</Emphasis> on apple fruit

Two antagonistic yeasts, Candida membranaefaciens and Pichia guilliermondii, were evaluated for the control of the blue mold of apple caused by Penicillium expansum. Dual culture, cell-free metabolite and volatile tests were used for in vitro assay. Yeast strains of two genera inhibited growth of P. expansum; inhibition varied from 30.27% to 44.19% in dual culture, from 79.40% to 90.57% in the volatile metabolite test, and from 72.99% to 88.77% in the cell-free metabolite test. Calcium chloride (2% w/v) significantly inhibited the growth of the pathogen P. expansum, but did not affect the colony-forming units (CFU) of the yeasts C. membranaefaciens and P. guilliermondii in potato dextrose broth. The concentration of yeast suspension influenced spore germination and germ tube growth of P. expansum in vitro, as well as disease incidence and lesion development in fruits. There were significant negative relationships between the suspension concentrations of the yeasts and the growth as well as infectivity of the pathogen. The addition of calcium resulted in lower spore germination rates and slower growth of germ tubes in vitro, as well as in lower disease incidences and smaller lesion diameters compared with treatments with yeast antagonists alone. When yeast cell suspensions reached a concentration of 10⁷ CFU ml^-1, growth of the pathogen was completely limited in vitro, and no infection was found in apple fruits treated with or without calcium. This article has been retracted because part of the data shown has already been published before, by different authors. An erratum to this article can be found at     

Evaluation of food additives and low-toxicity compounds as alternative chemicals for the control of Penicillium digitatum and Penicillium italicum on citrus fruit

The effectiveness of low-toxicity chemicals as possible alternatives to synthetic fungicides for the control of post-harvest green and blue moulds of citrus was evaluated. A preliminary selection of chemicals, mostly common food additives, was made through in vivo primary screenings with oranges artificially inoculated with Penicillium digitatum or P italicum. Selected compounds and mixtures were tested as heated solutions in small-scale trials. Immersion of artificially inoculated oranges or lemons for 120 s in solutions at 40.6 degrees C and natural pH of potassium sorbate (0.2 M), sodium benzoate (0.2 M) or mixtures (0.1 + 0.1 M) of potassium sorbate with sodium benzoate, sodium propionate or sodium acetate were the most effective organic acid salts tested and reduced green mould by 70-80% after 7 days of storage at 20 degrees C. The mixtures did not significantly enhance the effectiveness of potassium sorbate or sodium benzoate alone. These solutions were as effective as sodium carbonate or calcium polysulphide treatments and, in general, they were more effective on lemons than on oranges. Satisfactory control of green and blue moulds was obtained by dipping oranges for 150 s in solutions of sodium molybdate (24.2 mM) or ammonium molybdate (1.0 mM) at 48 or 53 degrees C, but not at 20 degrees C. At 53 degrees C, however, the effectiveness of hot water was not enhanced by either molybdate. Molybdenum salts at higher concentrations were phytotoxic and stained the fruit. At non-phytotoxic concentrations, the effectiveness of these solutions was more influenced by temperature than by concentration. In general, the inhibitory effects of all compounds tested were not fungicidal but fungistatic and not very persistent. In conclusion, potassium sorbate, sodium benzoate and ammonium molybdate, among the wide range of chemicals tested, were superior for the control of post-harvest Penicillium decay of citrus fruit.     

Control of Citrus Green and Blue Molds with Garlic Extracts

Water and ethanol extracts of garlic cloves were applied to artificially inoculated citrus fruits to test their efficacy in the control of Penicillium digitatum and P. italicum, the cause of citrus green and blue mold respectively. Extracts were tested either alone, or in combination with vegetable (sunflower) cooking oil or fruit wax at the rate of (0.1% v/v), using two orange cultivars (Valencia and Shamouti), and grapefruit. Treated fruits were stored at 10±1 °C, and 90–95% relative humidity for 30 days. All concentrations of extracts were more effective than the water control in inhibiting the growth and development of both pathogens, but were not as effective as the fungicide treatment (imazalil 500 ppm + quazatine 1000 ppm). A remarkable increase in the activity of garlic extracts was observed when extracts were mixed with oil. Consequently, the treatment comprising 1% extract plus oil was as effective (100% control) as the fungicide treatment in controlling both green and blue molds on Valencia oranges.     

Effect of ammonium carbonate and sodium bicarbonate on anthracnose of papaya

Colletotrichum gloeosporioides is the causal organism of anthracnose inCarica papaya L. (papaya, papaw). The effect of ammonium carbonate (3%) or sodium bicarbonate (2%) in aqueous solution or when incorporated into a wax formulation on anthracnose severity in inoculated or naturally infected fruits was examined. Both salts had significant effects, but that of ammonium carbonate was greater than that of sodium bicarbonate in controlling anthracnose. Ammonium carbonate (3%) incorporated into the wax formulation effectively reduced anthracnose incidence by 70% in naturally infected papaya and extended the storage life by maintaining the firmness, color and overall quality of the fruit in low temperature storage (13.5°C) and 95% r.h. for 21 days followed by 2 days under marketing conditions. The mode of action of ammonium carbonate on the control of anthroacnose appears to bevia complete inhibition of radial mycelial growth and conidia germination. http://www.phytoparasitica.org posting Aug. 14, 2002.     

Isolation of <Emphasis Type="Italic">Penicillium hirsutum</Emphasis> from spoiled,packaged asparagus spears in Japan

During storage studies on asparagus spears packed in plastic film and held at 15°C, a Penicillium fungus was associated with subsequent spoilage. On the basis of colony characteristics, the isolate was identified as Penicillium hirsutum Diercks. This report is the first of Penicillium hirsutum causing spoilage of asparagus spears in Japan. Inoculation tests showed that among all the Penicillium species tested in this study only the identified fungus was able to infect asparagus spears.     

Combination of hot water, <Emphasis Type="Italic">Bacillus subtilis</Emphasis> CPA-8 and sodium bicarbonate treatments to control postharvest brown rot on peaches and nectarines

The aim of this study was to evaluate the effect of hot water (HW), antagonists and sodium bicarbonate (SBC) treatments applied separately or in combination to control Monilinia spp. during the postharvest storage of stone fruit. Firstly, we investigated the effect of HW temperatures (55–70°C) and exposure times (20–60 s), seven antagonists at two concentrations (10⁷ or 10⁸ cfu ml⁻¹) and four SBC concentrations (1–4%). The selected treatments for brown rot control without affecting fruit quality were HW at 60°C for 40 s, SBC at 2% for 40 s and the antagonist CPA-8 (Bacillus subtilis species complex) at 10⁷ cfu ml⁻¹. The combinations of these treatments were evaluated in three varieties of peaches and nectarines artificially inoculated with M. laxa. When fruit were incubated for 5 d at 20°C, a significant additional effect to control M. laxa was detected with the combination of HW followed by antagonist CPA-8. Only 8% of the fruit treated with this combination were infected, compared to 84%, 52% or 24% among the control, CPA-8, and HW treatments, respectively. However, the other combinations tested did not show a significant improvement in effectiveness to control brown rot in comparison with applying the treatments separately. When fruit were incubated for 21 d at 0°C plus 5 d at 20°C, the significant differences between separated or combined treatments were reduced and generally the incidence of brown rot was higher than when fruit were incubated for 5 d at 20°C. Similar results were observed testing fruit with natural inoculum.     

Natural occurrence of citrinin in rice grains and its biocontrol byTrichoderma hamatum

Paddy rice was sampled from El-Sharkia, El-Gharbia, El-Dakahlia and Kafr El-Shekh governorates, Egypt. Of the 30 samples taken, ten were contaminated with the mycotoxin citrinin. An average of 6.79 × 10⁴ fungal spores per gram rice was found. The isolated fungi represented 47 species and 28 genera. The predominant genera wereAspergillus, Cladosporium andPenicillium. Aspergilli were represented by 22 species;Aspergillus niger andA. flavus had the highest occurrence.Penicillium viridicatum produced the highest amount of citrinin on glucose ammonium nitrate salts broth and rice grains, and hence this isolate was selected as a good producer of citrinin in this study. The presence ofTrichoderma hamatum reduced the amount of citrinin produced byP. viridicatum compared with its respective control. The excessive growth ofT. hamatum onP. viridicatum was increased with time. Viability ofP. viridicatum conidia decreased byT. hamatum with an increase in the incubation period. Chitinases and 1,3-β-glucanase enzyme activity ofT. hamatum increased with extending the incubation period onP. viridicatum mycelia up to maximum values at 72 and 84 h, respectively.T. hamatum led to a decrease in the production of citrinin byP. viridicatum on rice grains compared with the respective control values. http://www.phytoparasitica.org posting Dec. 19, 2004.     

Antifungal activity of sodium propylparaben alone or in combination with low doses of imazalil against Penicillium decay on citrus fruit

The performance of postharvest treatments with sodium propylparaben (SPP), alone or combined with low doses of the fungicide imazalil (IMZ), against citrus green (GM) and blue (BM) molds was evaluated on several citrus species and cultivars artificially inoculated with Penicillium digitatum and P. italicum, respectively, and incubated at 20 °C or cold-stored at 5 °C. Effectiveness of 100 mM SPP dips at 20 °C for 60 s was higher on oranges than on mandarins, with GM and BM incidence reductions of up to 60–90 % after 7 days at 20 °C. Irrespective of citrus cultivar and storage condition, SPP generally improved the curative action of 25 μl l^?1 IMZ to control Penicillium molds. In additional tests, 100 mM SPP dips at 20 °C for 60 s only prevented GM on ‘Valencia’ oranges inoculated 24 h after treatment when combined with IMZ. It can be concluded that postharvest SPP treatments show promise as an effective alternative to be considered in citrus postharvest disease control programs.     

Effects of inoculum density, leaf age, moisture, temperature, and wetness duration on black streak of edible burdock

Inoculum density, temperature, leaf age, and wetness duration were evaluated for their effects on the development of black streak (Itersonilia perplexans) on edible burdock (Arctium lappa L.) in a controlled environment. The effect of relative humidity (RH) on ballistospores production by I. perplexans was also evaluated. Symptoms of black streak on leaves increased in a linear fashion as the inoculum density of I. perplexans increased from 10² to 10⁶ ballistospores/ml. Rugose symptoms on young leaves were observed at densities of ≥10⁴ ballistospores/ml. Disease severity of I. perplexans in relation to leaf age followed a degradation curve when the leaves were inoculated with ballistospores. Disease severity was high in newly emerged leaves up to 5 days old, declined as leaf age increased to 29 days, and was zero when leaf age increased from 30 to 33 days. Disease development of edible burdock plants exposed to ballistospores of I. perplexans was evaluated at various combinations of temperature (10°, 15°, 20°, 25°C) and duration of leaf wetness (12, 24, 36, 48, and 72 h). Disease was most severe when plants were in contact with the ballistospore sources at 15° or 20°C. The least amount of disease occurred at 25°C regardless of wetness duration. Ballistospores required 24–36 h of continuous leaf wetness to cause visible symptoms by infection on edible burdock. Ballistospores production in infected lesions required at least 95.5% RH.     

Improved Control of Postharvest Decay of Pears by the Combination of Candida sake (CPA-1) and Ammonium Molybdate

ABSTRACT The potential enhancement of Candida sake (CPA-1) by ammonium molybdate to control blue and gray mold caused by Penicillium expansum and Botrytis cinerea, respectively, on Blanquilla pears was investigated. In laboratory trials, improved control of blue and gray molds was obtained with the application of ammonium molybdate (1, 5, 10, and 15 mM) alone or in combination with C. sake at 2 x 10(6) or 2 x 10(7) CFU ml(-1) on Blanquilla pears stored at 20 degrees C. In semicommercial trials at 1 degrees C for 5 months, the efficacy of C. sake at 2 x 10(6) CFU ml(-1) on reducing P. expansum and B. cinerea decay was enhanced more than 88% with the addition of ammonium molybdate 5 mM in the 1999-2000 season. In two seasons, the performance C. sake at 2 x 10(6) CFU ml(-1) plus ammonium molybdate was similar to or greater than that of C. sake at 2 x 10(7) CFU ml(-1). Similar control of blue mold was obtained on pears stored under low oxygen conditions. The preharvest application of ammonium molybdate did not reduce postharvest blue mold decay. The population of C. sake on pear wounds significantly decreased in the presence of ammonium molybdate 1 and 5 mM at 20 and 1 degrees C.     

Nutritional Enhancement of Biocontrol Activity of Candida sake (CPA-1) against Penicillium expansum on Apples and Pears

Pome fruits are poor in nitrogenous compounds and the addition of nitrogen can improve colonisation of the fruits by antagonists. Twenty-two nitrogenous compounds were evaluated for their effect on Candida sake (CPA-1) growth in vitro. Ten compounds that induced greater growth were applied with the antagonist to wounded fruits to evaluate their effect on enhancing control of Penicillium expansum. Calcium chloride and 2-deoxy-D-glucose were also tested. L-serine and L-aspartic acid enhanced biocontrol by C. sake against P. expansum on apples. On apples and pears, ammonium molybdate, calcium chloride and 2-deoxy-D-glucose improved the capacity of the antagonist to control P. expansum. The addition of ammonium molybdate at 1 mM allowed C. sake to be used on apples and pears at a lower concentration without diminishing control. Similar results were observed with the addition of calcium chloride to the antagonist. 2-deoxy-D-glucose at 6 and 18 mM enhanced biocontrol on pears by over 81%, but on apples the improvement of biocontrol was observed only at 6 mM. In cold storage, the combination of ammonium molybdate and C. sake completely eliminated the incidence of blue mould on pears, and reduced its severity and incidence by more than 80% on apples.     

Molecular Approaches to Assist the Screening and Monitoring of Postharvest Biocontrol Yeasts

Nineteen yeast isolates obtained from the surface of several fruits and vegetables grown in Southern Italy and Israel were compared by molecular analysis using arbitrarily primed polymerase chain reaction (AP-PCR) and random amplified polymorphic DNA technique (RAPD-PCR). Genetic analysis made it possible to distinguish between closely-related genitically different strains which had the same morphological characteristic, and to discard isolates which were genetically identical. Following PCR characterisation, 6 isolates were selected and tested for their biocontrol activity against major postharvest pathogens (Penicillium digitatum on grapefruit, Botrytis cinerea, Rhizopus stolonifer and Aspergillus niger on table grape and B. cinerea and R. stolonifer on cherry tomato). All the isolates showed a good biocontrol efficacy on both wounded and non-wounded fruits. Furthermore, the preharvest application of the most effective antagonist (LS15) on table grape resulted in a significant reduction in grey mold ranging from 28.3% to 38.2% compared to the untreated control. The RAPD-PCR technique was also useful for identifying and monitoring the survival of the antagonist after field application.     

<Emphasis Type="Italic">Ulocladium atrum</Emphasis> as a biological control agent for white mold of bean caused by<Emphasis Type="Italic">Sclerotinia sclerotiorum</Emphasis>

Field studies were conducted near Lethbridge, Alberta, Canada, in 2001, 2004 and 2005 to determine the efficacy of the antagonistic fungusUlocladium atrum for control of white mold of bean caused bySclerotinia sclerotiorum. Results of the 3 years of field trials showed that, compared with the untreated control, foliar application of a spore suspension ofU. atrum (300 ml m⁻² of 10⁶ spores ml⁻¹ suspension) significantly reduced incidence and severity of white mold, increased seed yield and reduced contamination of bean seed by sclerotia ofS. sclerotiorum. The level of control of white mold observed in the treatment ofU. atrum was similar to that of the mycoparasitic fungusConiothyrium minitans, but lower than the fungicide treatments of Ronilan (vinclozolin) at the rate of 1200 g ha⁻¹ per application in 2001, or Lance (boscalid) at the rate of 750 g ha⁻¹ per application in 2004 and 2005. The potential for use ofU. atrum as a biological control agent for sclerotinia diseases is discussed. http://www.phytoparasitica.org posting Nov. 12, 2006.     

First report of whisker mold,a postharvest disease on citrus caused by <Emphasis Type="Italic">Penicillium ulaiense</Emphasis> (in Japan)

In March 2006, stored fruits of the medium-to-late-ripening citrus variety Shiranuhi ([Citrus unshiu × C. sinensis] × C. reticulata) were found to have a disease similar to blue mold. The fungus causing this disease differed distinctly from the well-known, blue mold agent, Penicillium italicum, because it formed whisker-like coremia measuring 1–8 mm. On the basis of the morphological characteristics and the phylogenetic analysis based on the nucleotide sequence of the β-tubulin gene, the fungus was identified as P. ulaiense. This is the first report of citrus whisker mold caused by P. ulaiense in Japan.     

Studies on in vitro Growth and Pathogenicity of European Fusarium Fungi

The effect of temperature on the in vitro growth rates and pathogenicity of a European Fusarium collection consisting of isolates of Fusarium graminearum, F. culmorum, F. avenaceum, F. poae and Microdochium nivale was examined. Irrespective of geographic origin, the optimum temperature for the growth of F. graminearum, F. culmorum and F. poae was 25 °C, while that for F. avenaceum and M. nivale was 20 °C. In general, the growth rates of F. graminearum, F. culmorum and F. poae increased between 10 and 25 °C and those of F. avenaceum and M. nivale increased between 10 and 20 °C. Pathogenicity tests were carried out by examining the effect of the five species on the in vitro coleoptile growth rate of wheat seedlings (cv. Falstaff). Irrespective of geographic origin, the temperature at which F. avenaceum, F. culmorum and F. graminearum caused the greatest retardation in coleoptile growth ranges 20–25 °C (>89.3% reduction), whilst for F. poae and M. nivale it was 10–15 °C (>45.6% retardation), relative to uninoculated control seedlings. In general, F. culmorum and F. graminearum were the most pathogenic of the five species, causing at least a 69% reduction in coleoptile growth at 10, 15, 20 and 25 °C. General linear model analysis (GLIM) showed that species accounted for 51.3–63.4% of the variation in isolate growth and from 19.5% to 44.3% of the variation in in vitro pathogenicity. Country of origin contributed from 22.6% to 51.9% to growth rate variation and from 0.73% to 7.61% to pathogenicity variation. The only significant correlation between in vitro growth and pathogenicity was that observed for M. nivale at 15 °C (r = -0.803, P < 0.05).     

Germination of <Emphasis Type="Italic">Gibberella zeae</Emphasis> ascospores as affected by age of spores after discharge and environmental factors

The effects of age of ascospores (0–18 days after discharge), photon flux density (0–494 mol m^–2 s^–1 PAR), temperature (4–30 °C), frost (–15 °C for 30 min), relative humidity (RH; 0–100%), pH (2.5–6.5) and dryness (0 and 53% RH for up to 40 min) on the germination of the ascospores of the mycotoxin-producing fungus Gibberella zeae (anamorph Fusarium graminearum) were studied. Freshly discharged ascospores germinated within 4 h at 20 °C and 100% RH. The rate of germination and the percentage of viable ascospores decreased over time after the spores were discharged from perithecia. The time course of ascospore germination was not significantly affected by photon flux density. The period of time required to obtain 50% germinated ascospores at 100% RH was 26.90 h at 4 °C, 10.40 h at 14 °C, 3.44 h at 20 °C and 3.31 h at 30 °C. There was no significant effect of frost on the percentage of viable ascospores. A small percentage (6.6 ± 3.8%) of the ascospores germinated at 53% RH. At RH 84% and 20 °C almost 100% of the freshly discharged ascospores germinated. The time course of ascospore germination was affected by pH. The maximum rate of ascospore germination was estimated to be at pH 3.76. Ascospores lost their ability to germinate following exposure to 0% RH almost instantaneously. No germinating spores were detected after an incubation period of 1 min at 0% RH. Incubating the ascospores at 53% RH decreased the percentage of viable spores from 93 to 6% within 10 min. The data demonstrate that age of spores, relative humidity, temperature and pH, but not photon flux density, are key factors in germination of G. zeae ascospores.