Viability and release of Neonectria ditissima ascospores on apple fruit in Brazil

During European canker monitoring in an apple experimental orchard, 14 mummified fruit (two and three trees with 10 and four positive records in 2018 and 2019, respectively) showed perithecia. Perithecium production on apple fruit, confirmation of pathogenicity of Neonectria ditissima isolated from mummified fruit, and ascospore release from fruit tissues has rarely been reported, and their role in the epidemiology of European canker has been largely overlooked. Thus, the objectives of our study were to (a) prove the presence of both conidia and ascospores of N. ditissima in mummified fruit in an experimental field, confirming pathogenesis in different apple cultivars, and (b) monitor production of the two types of inoculum in infected apple fruit over time. Canker incidence in this orchard was 47% of trees with symptoms in 2018 and 48% in 2019. Molecular and morphological tests confirmed that the fungus detected in the mummified apple fruit was N. ditissima. Apple fruit with sporodochia and perithecia washed immediately after collection from the orchard showed conidia but no ascospores of N. ditissima. However, after 4 days’ incubation, perithecia on mummified fruit showed many ascospore cirri. Koch's postulates were fulfilled on apple plants and mature fruit. Fruit inoculated with N. ditissima released spores for over a year under Brazilian field conditions. The release of both spore types peaked in May (Brazilian leaf fall) and October (spring); release of conidia also peaked in February (early harvest). These results support our hypothesis that fruit can serve as primary inoculum for European canker in Brazilian apple orchards.     

Growth and sporulation of <Emphasis Type="Italic">Stemphylium vesicarium</Emphasis>, the causal agent of brown spot of pear,on herb plants of orchard lawns

The inoculum sources of ascospores of Pleospora allii and of conidia of its anamorph Stemphylium vesicarium were investigated in relation to the brown spot disease epidemiology on pear. Dead and living leaves of three pear varieties (Abate Fétel, Conference and William), seven grasses (Poa pratensis, Festuca rubra, Festuca ovina, Lolium perenne, Digitaria sanguinalis and Setaria glauca) and Trifolium repens, which are used in pear orchard lawns, were inoculated with conidia of Stemphylium vesicarium virulent on pear and incubated under controlled-environment. Stemphylium vesicarium was always re-isolated from dead leaves of the considered plants, but not from symptomless green or yellowish living leaves. The fungus was occasionally re-isolated from leaf segments showing unspecific necrosis. Inoculation of pear leaves with isolates from grasses demonstrated that the fungus did not lose pathogenicity. Pseudothecia, ascospores and conidia were produced on all the dead inoculated leaves; differences between specimens were found for phenology of pseudothecia, their density and size, and for the number of conidia produced. Pseudothecia were produced faster in the lawn species than in pear leaves, and their density was higher, especially for S. glauca, L. perenne and P. pratensis. Ascospore maturation and ejection was more concentrated for the pseudothecia developed on pear leaves than for those on F. ovina and S. glauca. All the lawn species produced more conidia than pear leaves.     

苹果炭疽叶枯病病原Glomerella cingulata及其侵染过程

为明确苹果炭疽叶枯病病原菌围小丛壳Glomerella cingulata的侵染致病特征,在分离获得该病原菌的基础上,采用形态学观察、ITS序列分析和致病性测定对其进行了鉴定,并利用光学和扫描电子显微镜对病原菌在嘎啦苹果叶片上的侵染过程进行了研究.结果表明,在陕西咸阳地区分离获得的9株病原菌均为围小丛壳G.cingulata.25 ℃下接种9 h后,分生孢子中间产生隔膜,双胞化,并萌发产生芽管和附着胞;24 h后分生孢子的2个细胞均可萌发并形成芽管及附着胞,部分芽管顶端可产生次级分生孢子;48 h后次级分生孢子萌发形成附着胞;72 h后,附着胞下形成的侵染钉可直接入侵寄主,在表皮细胞内形成初生菌丝和次生菌丝,此时叶片表面已出现褐色斑点.接种7 d后叶片病斑处出现分生孢子盘和子囊壳.表明陕西省近年出现的苹果炭疽叶枯病病原菌为围小丛壳G.cingulata,该病菌在嘎啦叶片上的一些特殊侵染行为可能是导致该病害易在短时间内暴发的重要原因.     

The relative importance of conidia and ascospores as primary inoculum of Venturia inaequalis in a southeast England orchard

Apple scab, caused by Venturia inaequalis, can lead to large losses of marketable fruit if left uncontrolled. The disease appears in orchards during spring as lesions on leaves. These primary lesions are caused by spores released at bud burst from overwintering sources; these spores can be sexually produced ascospores from the leaf litter or asexual conidia from mycelium in wood scab or within buds. The relative importance of conidia and ascospores as primary inoculum were investigated in an orchard in southeast England, UK. Potted trees not previously exposed to apple scab were placed next to (c. 1 m) orchard trees to trap air‐dispersed ascospores. Number and position of scab lesions were assessed on the leaves of shoots from both the potted trees (infection by airborne ascospores) and neighbouring orchard trees (infection by both ascospores and splash‐dispersed, overwintered conidia). The distribution and population similarity of scab lesions were compared in the two tree types by molecular analysis and through modelling of scab incidence and count data. Molecular analysis was inconclusive. Statistical modelling of results suggested that conidia may have contributed approximately 20–50% of the primary inoculum in early spring within this orchard: incidence was estimated to be reduced by 20% on potted trees, and lesion number by 50%. These results indicate that, although conidia are still a minority contributor to primary inoculum, their contribution in this orchard is sufficient to require current management to be reviewed. This might also be true of other orchards with a similar climate.     

Effects of inoculum density,leaf wetness duration and nitrate concentration on the occurrence of lettuce leaf spot

In Ehime Prefecture, Japan, lettuce leaf spot (Septoria lactucae) caused huge losses in marketable lettuce yields. To explore potential measures to control disease outbreaks, the effects of inoculum density, leaf wetness duration and nitrate concentration on the development of leaf spot on lettuce (Lactuca sativa) were evaluated. Conidia were collected from diseased plants in an infested field by single-spore isolation and were used to inoculate potted lettuce plants with different conidial concentrations. Lesions developed on inoculated lettuce plants at inoculum concentrations from 10⁰ to 10⁶ conidia/ml. The disease was more severe when the inoculum exceeded 10² conidia/ml, and severity increased with increasing concentrations. Assessment of the relationship between disease development and the duration of postinoculation leaf wetness revealed that symptoms appeared when the inoculated plants remained wet for 12 h or longer. The number of lesions and total nitrogen content in the lettuce leaves both increased when nitrate was applied.     

Sources and seasonal dynamics of inoculum for brown spot disease of pear

The dynamics of the production of Stemphylium vesicarium conidia and Pleospora allii ascospores from different inoculum sources on the ground were compared in a model system of a wildflower meadow mainly composed of yellow foxtail, creeping cinquefoil and white clover. The meadow was either inoculated (each October) or not inoculated with a virulent strain of S. vesicarium, and either covered or not covered with a litter of inoculated pear leaves. Spore traps positioned a few centimetres above the ground were exposed for 170 7-day periods between October 2003 and December 2006. Ascospores and conidia were trapped in 46 and 25% of samples, respectively. Ascospore numbers trapped from the pear leaf litter were about five times higher than those from the meadow, while conidial numbers were similar from the different inoculum sources. The ascosporic season was very long, with two main trapping periods: December–April, and August–October; the former was most important for the leaf litter, the latter for the meadow. The conidial season lasted from April to November, with 92% of conidia caught between July and September. The fungus persistently colonized the meadow: the meadow inoculated in early October 2003 produced spores until autumn 2006. The present work demonstrates that orchard ground is an important source of inoculum for brown spot of pear. Thus, it is important to reduce inoculum by managing the orchard ground all year long.     

Population structure of Valsa ceratosperma, causal fungus of Valsa canker, in apple and pear orchards

On the basis of mycelial compatibility, the population structure of Valsa ceratosperma, the causal fungus of Valsa canker on broadleaf trees, was investigated in apple and pear orchards in Japan. Field strains of V. ceratosperma from a single canker on trunks or scaffold limbs belonged to different mycelial compatibility groups. Thus, the population structure of this fungus was complex in most orchards. Because mycelia of strains originating from different conidia from the same pycnidium were compatible, infection by this fungus is thought to be ascospores.     

Selection and orchard testing of antagonists suppressing conidial production by the apple scab pathogen <Emphasis Type="Italic">Venturia inaequalis</Emphasis>

Apple scab caused by Venturia inaequalis is a major disease in apple production. Epidemics in spring are initiated by ascospores produced on overwintering leaves whereas epidemics during summer are driven by conidia produced on apple leaves by biotrophic mycelium. Fungal colonisers of sporulating colonies of V. inaequalis were isolated and their potential to reduce the production of conidia of V. inaequalis was evaluated on apple seedlings under controlled conditions. The four most effective isolates of the 63 screened isolates were tested subsequently under Dutch orchard conditions in 2006. Repeated applications of conidial suspensions of Cladosporium cladosporioides H39 resulted in an average reduction of conidial production by V. inaequalis of approximately 40%. In 2007, applications of conidial suspensions of C. cladosporioides H39 reduced conidial production by V. inaequalis by 69% on August 6 and by 51% on August 16, but no effect was found on August 20. However, viability of available conidia of C. cladosporioides H39 was low at the end of the experiment. Epiphytic and endophytic colonisation by Cladosporium spp. of leaves treated during the experiment with C. cladosporioides H39 was significantly higher than on control leaves sampled 6 weeks after the last application. It is concluded that C. cladosporioides H39 has promising potential as a biological control agent for apple scab control. More information is needed on the effect of C. cladosporioides H39 on apple scab epidemics as well as on mass production, formulation and shelf life of conidia of the antagonist.     

Aloe ring spot,a new disease of aloe caused by <Emphasis Type="Italic">Haematonectria haematococca</Emphasis> (Berk. &; Broome) Samuels &; Nirenberg (anamorph: <Emphasis Type="Italic">Fusarium</Emphasis> sp.)

A ring spot disease of Aloe vera was found on leaves of potted seedlings of Aloe vera in Hachijojima and Chichijima Islands, Tokyo. From tissue of ring spot lesions, a fungus producing Fusarium-type conidia was consistently isolated. After 1 month, reddish perithecia of nectriaceous fungus had formed on the colonies of this isolate on PDA. These nectriaceous and Fusarium fungi were identified as Haematonectria haematococca and Fusarium sp., respectively. From a single ascospore isolation, the former was confirmed to be the teleomorph of the Fusarium sp. Typical ring spot lesions were reproduced by artificial inoculations using single ascospore and single conidium isolates. Inoculations of five species of genus Aloe revealed that they were highly susceptible except for A. arborescens. This is the first report of a disease on Aloe caused by H. haematococca (anamorph: Fusarium sp.) in Japan, and it was named aloe ring spot.     

Invasion and colonization of mature apple fruit by Erwinia amylovora tagged with bioluminescence genes

Invasion and colonization of mature apple fruit by a transformant of Erwinia amylovora tagged with bioluminescence genes from Vibrio fischeri was examined. The transformant was deposited on cut surfaces of fruit stems, wounds on the shoulders and calyces, injured fruit-bearing twigs of harvested apple fruit, and cut fruit flesh. After incubation in closed stainless steel or plastic boxes at 25°C, fruit were periodically observed with a two-dimensional luminometer. The presence of the transformant in luminous areas was confirmed by isolating it on selective media. E. amylovora, when deposited in fruit stems: (1) can invade mature as well as immature apple fruit; (2) vertically and horizontally spreads and colonizes along vascular bundles, increasing its population; (3) reaches the calyx end and the flesh just under the exocarp within 3–4 days after inoculation; (4) when deposited on cut fruit flesh, irrespective of its maturity, can easily increase its population and survive 2–4 weeks or more at 25°C; and (5) even at the time of fruit maturation, can migrate within twigs rapidly and reaches the abscission layers between fruit-bearing twigs and fruit stems.     

A dynamic model simulating infection of apple leaves by Venturia inaequalis

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

Effect of interrupted leaf wetness periods on suppression of sporulation ofBotrytis allii andB. cinerea by antagonists on dead onion leaves

Saprophytic antagonists were evaluated for suppression of sporulation ofBotrytis allii andB. cinerea on artificially killed segments of onion leaves that were pre-inoculated with the pathogens. During incubation of the antagonisttreated leaf segments in moist chambers, periods of leaf wetness and leaf dryness were alternated to simulate conditions in the field. Interruption of humid conditions with dry periods had a differential effect on antagonists.Alternaria alternata, Chaetomium globosum, Ulocladium atrum andU. chartarum suppressed sporulation ofB. allii almost completely under continuously wet conditions, and when the leaf wetness periods were interrupted with drying periods of 9h imposed 16, 40, and 64 h after the antagonists were applied. When leaf wetness was interrupted 16 h after antagonist application, the number of conidia ofB. allii produced cm^–2 leaf surface after eight days was under the detection limit of 5.2 × 10³ conidia on leaves treated with these antagonists compared to 3.7 × 10⁵ conidia on leaves that were not treated. On the other hand,Gliocladium roseum, G. catenulatum andSesquicillium candelabrum, all highly efficient under continuously wet conditions, were of low to moderate efficiency when leaf wetness periods had been interrupted 16 h after application of the antagonists. The antagonists showed the same differentiation and sensitivity to interrupted wetness periods when tested withB. cinerea.     

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

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

Multiple Alternaria species groups are associated with leaf blotch and fruit spot diseases of apple in Australia

Leaf blotch and fruit spot of apple caused by Alternaria species occur in apple orchards in Australia. However, there is no information on the identity of the pathogens and whether one or more Alternaria species cause both diseases in Australia. Using DNA sequencing and morphological and cultural characteristics of 51 isolates obtained from apple leaves and fruit with symptoms in Australia, Alternaria species groups associated with leaf blotch and fruit spot of apples were identified. Sequences of Alternaria allergen a1 and endopolygalacturonase gene regions revealed that multiple Alternaria species groups are associated with both diseases. Phylogenetic analysis of concatenated sequences of the two genes resulted in four clades representing A. arborescens and A. arborescens‐like isolates in clade 1, A. tenuissima/A. mali isolates in clade 2, A. alternata/A. tenuissima intermediate isolates in clade 3 and A. longipes and A. longipes‐like isolates in clade 4. The clades formed using sequence information were supported by colony characteristics and sporulation patterns. The source of the isolates in each clade included both the leaf blotch variant and the fruit spot variant of the disease. Alternaria arborescens‐like isolates were the most prevalent (47%) and occurred in all six states of Australia, while A. alternata/A. tenuissima intermediate isolates (14%) and A. tenuissima/A. mali isolates (6%) occurred mostly in Queensland and New South Wales, respectively. Implications of multiple Alternaria species groups on apples in Australia are discussed.     

Fitness Cost Associated with Loss of the AvrLm4 Avirulence Function in Leptosphaeria maculans (Phoma Stem Canker of Oilseed Rape)

Near-isogenic isolates of Leptosphaeria maculans differing at the AvrLm4 avirulence locus (AvrLm4 or avrLm4) were produced in vitro. Methods for inoculation of leaves of oilseed rape with ascospores or conidia were compared. The ‘ascospore shower’ inoculation was the most efficient method for use when inoculum is limited (e.g. ascospores produced in vitro). It was used in controlled environments to compare fitness of AvrLm4 and avrLm4 isolates at 5, 10, 15, 20 or 25 °C on leaves of oilseed rape cultivars Eurol and Darmor lacking the resistance gene Rlm4, which corresponds to AvrLm4. At all temperatures tested, AvrLm4 ascospores produced more lesions than avrLm4 ascospores. The diameters of lesions produced by AvrLm4 ascospores were greater than those of lesions produced by avrLm4 ascospores. At 15–20 °C, more lesions initiated by AvrLm4 ascospores produced pycnidia than did lesions initiated by avrLm4 ascospores. However, there were no differences between AvrLm4 and avrLm4 isolates in incubation period (from inoculation to appearance of lesions) or rate of mycelial growth in leaves from lesions towards the stems. In field experiments with winter oilseed rape cultivars lacking Rlm4, the frequency of AvrLm4 isolates increased from 5.7% at the phoma leaf lesion stage (autumn) to 20.5% at the stem canker stage (summer) during 2002/2003 and from 7.9 to 11.5% during 2003/2004 growing seasons. Results of controlled environment and field experiments indicate that avrLm4 isolates have a fitness cost compared to AvrLm4 isolates.     

Pathogenicity,colony morphology and diversity of isolates of <Emphasis Type="Italic">Guignardia citricarpa</Emphasis> and <Emphasis Type="Italic">G. mangiferae</Emphasis> isolated from <Emphasis Type="Italic">Citrus</Emphasis> spp.

In the present study, the pathogenicity of 36 isolates of Guignardia species isolated from asymptomatic ‘Tahiti’ acid lime fruit peels and leaves, ‘Pêra-Rio’ sweet orange leaves and fruit peel lesions, and a banana leaf were characterized. For pathogenicity testing, discs of citrus leaves colonized by Phyllosticta citricarpa under controlled laboratory conditions were kept in contact with the peels of fruit that were in susceptible states. In addition, pathogenicity was related to morphological characteristics of colonies on oatmeal (OA) and potato dextrose agar (PDA). This allowed the morphological differentiation between G. citricarpa and G. mangiferae. Polymerase chain reactions (PCRs) were also used to identify non-pathogenic isolates based on primers specific to G. citricarpa. A total of 14 pathogenic isolates were detected during pathogenicity tests. Five of these were obtained from leaf and fruit tissues of the ‘Tahiti’, which until this time had been considered resistant to the pathogen. Given that the G. citricarpa obtained from this host was pathogenic, it would be more appropriate to use the term insensitive rather than resistant to categorize G. citricarpa. A non-pathogenic isolate was obtained from lesions characteristic of citrus black spot (CBS), indicating that isolation of Guignardia spp. under these conditions does not necessarily imply isolation of pathogenic strains. This also applied to Guignardia spp. isolates from asymptomatic citrus tissues. Using fluorescent amplified fragment length polymorphism (fAFLP) markers, typically pathogenic isolates were shown to be more closely related to one another than to the non-pathogenic forms, indicating that the non-pathogenic isolates display higher levels of genetic diversity.     

Characterization of monilia disease caused by Monilinia linhartiana on quince in southern Spain

This study elucidates the aetiology and epidemiology of monilia disease of quince caused by the fungus Monilinia linhartiana in Spain. Disease incidence and the dynamics of apothecial development and ascospore discharge were quantified and the pathogen was characterized using morphological and molecular methods. The pathogen did not produce conidia or apothecia on agar media but produced conidia on leaves showing symptoms and apothecia on mummified young quince fruit. Monilinia linhartiana was not pathogenic on ripe quince fruit but was readily isolated from developing, mummified fruit (pseudosclerotia). Phylogenetic analysis based on 5·8S‐ITS region sequences placed M. linhartiana in the Disjuntoriae section of Monilinia species infecting rosaceous hosts. Studies during 2004–2008 in four commercial orchards in southern Spain determined two major infection periods for the disease. The first coincided with the unfolding of the first leaves and resulted in leaf blotch and shoot blight. The second coincided with flowering and led to mummification of developing young fruit. Foliar infection was apparently initiated by airborne ascospores produced on pseudosclerotia that overwintered on the soil surface, while flower infection was probably initiated by conidia produced on leaf lesions. Incidence of diseased shoots ranged from 1 to 91% and was correlated with calculated inoculum potential, based on the density and maturity of apothecia formed on pseudosclerotia. This epidemiological study has made it possible to characterize the life cycle of monilia disease on quince in southern Spain, which will help the development of new control strategies.     

Relating plant and pathogen development to optimise fungicide control of phoma stem canker (<Emphasis Type="Italic">Leptosphaeria maculans</Emphasis>) on winter oilseed rape (<Emphasis Type="Italic">Brassica</Emphasis><Emphasis Type="Italic">napus</Emphasis>)

In winter oilseed rape experiments at Rothamsted in 2000/01 to 2002/03 growing seasons, the severity of phoma stem canker epidemics in summer depended on the timing of phoma leaf spot epidemics in the previous autumn, and hence on the timing of Leptosphaeria maculans ascospore release. The first major release of L. maculans ascospores was earlier in 2000 (26 September) and 2001 (18 September) than in 2002 (21 October). Consequently, the autumn phoma leaf spot epidemic was also earlier in 2000 and 2001 than in 2002. The resulting stem canker epidemics were severe by harvest (July) in 2001 and 2002 but not in 2003. No correlation was found between the severity or duration of phoma leaf spotting (lesion days or lesion °C-days) and the subsequent severity of phoma stem canker epidemics. Rates of leaf production and loss were similar in the three growing seasons. Out of ca. 25 leaves produced on plants during each season, leaf numbers 10–14 generally remained on plants for the longest. Treatment with flusilazole + carbendazim in autumn decreased the severity of phoma leaf spotting for several weeks after treatment, decreased the severity of stem canker the following summer and increased yield significantly in 2001 and 2002 but not in 2003. The most effective timings for flusilazole + carbendazim application were when leaves 7–11 were present on most plants and at least 10% of plants were affected by phoma leaf spot. Two half-dose applications of fungicide reduced phoma stem canker and increased yield more than a single full dose application when phoma leaf spot epidemics were early (<800 °C-days after sowing).     

Effect of temperature,relative humidity,leaf wetness and leaf age on <Emphasis Type="Italic">Spilocaea oleagina</Emphasis> conidium germination on olive leaves

The effects of temperature, relative humidity (RH), leaf wetness and leaf age on conidium germination were investigated for Spilocaea oleagina, the causal organism of olive leaf spot. Detached leaves of five ages (2, 4, 6, 8 and 10 weeks after emergence), six different temperatures (5, 10, 15, 20, 25 and 30°C), eight wetness periods (0, 6, 9, 12, 18, 24, 36 and 48 h), and three RH levels (60, 80 and 100%) were tested. Results showed that percentage germination decreased linearly in proportion to leaf age (P < 0.001), being 58% at 2 weeks and 35% at 10 weeks. A polynomial equation with linear term of leaf age was developed to describe the effect of leaf age on conidium germination. Temperature significantly (P < 0.001) affected frequencies of conidium germination on wet leaves held at 100% RH, with the effective range being 5 to 25°C. The percent germination was 16.1, 23.9, 38.8, 47.8 and 35.5% germination at 5, 10, 15, 20 and 25°C, respectively, after 24 h. Polynomial models adequately described the frequencies of conidium germination at these conditions over the wetness periods. The rate of germ tube elongation followed a similar trend, except that the optimum was 15°C, with final mean lengths of 175, 228, 248, 215 and 135 μm at 5, 10, 15, 20 and 25°C, respectively after 168 h. Polynomial models satisfactorily described the relationships between temperature and germ tube elongation. Formation of appressoria, when found, occurred 6 h after the first signs of germination. The percentage of germlings with appressoria increased with increasing temperature to a maximum of 43% at 15°C, with no appressoria formed at 25°C after 48 h of incubation. Increasing wetness duration caused increasing numbers of conidia to germinate at all temperatures tested (5–25°C). The minimum leaf wetness periods required for germination at 5, 10, 15, 20 and 25°C were 24, 12, 9, 9 and 12 h, respectively. At 20°C, a shorter wetness period (6 h) was sufficient if germinating conidia were then placed in 100% RH, but not at 80 or 60%. However, no conidia germinated without free water even after 48 h of incubation at 20°C and 100% RH. The models developed in this study should be validated under field conditions. They could be developed into a forecasting component of an integrated system for the control of olive leaf spot.