Evaluation of Infection Processes and Resulting Disease Caused by Dendryphion penicillatum and Pleospora papaveracea on Papaver somniferum

ABSTRACT Two pathogenic fungi of opium poppy, Pleospora papaveracea and Dendryphion penicillatum, were isolated from field material in Beltsville, MD. The processes of infection by these two fungi were studied to determine the optimal environmental conditions for infection. Both fungi formed appressoria capable of penetrating directly through the plant epidermal layer. Of the two fungi, P. papaveracea was more aggressive, causing more rapid necrosis. Appressorial formation by P. papaveracea occurred as early as 4 h after application of a conidial suspension to poppy leaves. P. papaveracea formed more appressoria than did D. penicillatum, especially at cool temperatures (7 to 13 degrees C). In greenhouse studies, P. papaveracea caused more damage to opium poppy than did D. penicillatum when applied in 10% unrefined corn oil. In the field, P. papaveracea was more consistent in its effects on opium poppy from a local seed source designated Indian Grocery. P. papaveracea caused higher disease ratings, more stem lesions, and equal or greater yield losses than did D. penicillatum on Indian Grocery. The late-maturing opium poppy variety White Cloud was severely damaged by disease, regardless of formulation or fungal treatment. P. papaveracea was the predominant fungus isolated from poppy seed capsules and the only fungus reisolated from the field the following year. These studies provide a better understanding of the infection process and the differences between these two pathogenic fungi and will be beneficial for the development of the fungi as biological control agents.     

Nep1 Protein from Fusarium oxysporum Enhances Biological Control of Opium Poppy by Pleospora papaveracea

ABSTRACT The fungus Pleospora papaveracea and Nep1, a phytotoxic protein from Fusarium oxysporum, were evaluated for their biocontrol potential on opium poppy (Papaver somniferum). Four treatments consisting of a control, P. papaveracea conidia, Nep1 (5 mug/ml), and P. papaveracea conidia plus Nep1 (5 mug/ml) were used in detached-leaf and whole-plant studies. Conidia of P. papaveracea remained viable for 38 days when stored at 20 or 4 degrees C. Nep1 was stable in the presence of conidia for 38 days when stored at 4 degrees C or for 28 days at 20 degrees C. The presence of Nep1 did not affect conidia germination or appressoria formation. Nep1 was recovered from drops applied to opium poppy leaves in greenhouse and field studies 24 h after treatment. Opium poppy treated with the combination of Nep1 and P. papaveracea had higher necrosis ratings than the other treatments. There were changes in the intercellular protein profiles, determined by sodium dodecyl sulfate gel electrophoresis and silver staining, due to application of treatments; the most intense occurred in response to the combination of Nep1 and P. papaveracea. The combination of Nep1 and P. papaveracea enhanced the damage caused to opium poppy more than either component alone.     

Development of the teleomorph of Ascochyta rabiei on culture media

Ascochyta blight caused by Didymella rabiei (anamorph: Ascochyta rabiei) is an important foliar disease of chickpea in many countries. The fungus is heterothallic and requires the pairing of two compatible mating types for the teleomorph to develop. In nature, the teleomorph only develops on chickpea debris that overwinters on the soil surface in the presence of both mating types. When natural and synthetic agar media were seeded with conidial suspensions of compatible isolates of D. rabiei from Spain and the United States and incubated under favourable conditions for teleomorph development, the teleomorph only developed on 2?% water agar amended with powdered chickpea stems or hot water extracts of chickpea stems, but not on 14 other natural or synthetic media. Ascospore isolates of D. rabiei from pseudothecia that developed on agar media were indistinguishable in cultural and morphological characteristics from isolates of the fungus from chickpea. Production of pseudothecia and ascospores on the best culture medium was always lower than on stem pieces of chickpea straw used as a control treatment. Ascospores discharged from pseudothecia that developed on powdered chickpea stem media onto chickpea seedlings were pathogenic, inducing symptoms identical to those caused by ascospores from chickpea stem pieces or conidia from a chickpea isolate of the fungus. This is the first report of the teleomorph of D. rabiei developing on culture media.     

The Occurrence of Mycosphaerella graminicola and its Anamorph Septoria tritici in Winter Wheat during the Growing Season

The disease septoria tritici blotch of wheat is initiated by ascospores of the teleomorph Mycosphaerella graminicola or pycnidiospores of the anamorph Septoria tritici. We report for the first time the presence of the teleomorph, M. graminicola, in Denmark. With the objective of elucidating the importance of the teleomorph for the development of septoria tritici blotch, data on the occurrence of fruit bodies of the anamorph (pycnidia) and the teleomorph (pseudothecia) stages were collected over three growing seasons. Pseudothecia were present in the springs, however, high numbers of pseudothecia compared to pycnidia were not observed until July, too late to influence the epidemic. On an individual leaf layer, pycnidia were observed well before pseudothecia. As the leaves aged, progressively higher proportions of fruit bodies were observed to be pseudothecia. The period from the appearance of pycnidia to detection of pseudothecia was estimated as 29–53 days. At harvest, high proportions of sporulating fruit bodies in the crop were pseudothecia, suggesting that the primary source of inoculum for new emerging wheat crops in autumn is likely to be ascospores.     

Alternative hosts and plant tissues for the survival,sporulation and spread of the Ascochyta blight pathogen of chickpea

Various crop and weed species were infected naturally by Didymella rabiei (anamorph: Ascochyta rabiei) in blight-affected chickpea fields in the Palouse region of eastern Washington and northern Idaho, USA. The fungus was isolated from asymptomatic plants of 16 species commonly found in commercial crops in this region. Isolates of the pathogen from crop and weed species were pathogenic to chickpea and indistinguishable in cultural and morphological characteristics from isolates of D. rabiei from chickpea. Both mating types of D. rabiei were isolated from eight naturally infected plant species. Chickpeas were infected by D. rabiei when plants emerged through infested debris of seven crop and weed species. The teleomorph developed on overwintered tissues of seven plant species infected naturally by D. rabiei in a blight screening nursery and on debris of wheat, white sweet clover and pea inoculated with ascospores of D. rabiei or conidia of two compatible isolates of the pathogen. Didymella rabiei naturally infected 31 accessions of 12 Cicer spp. and the teleomorph developed on the overwintered debris of all accessions, including those of three highly resistant perennial species. The fungus developed on the stem and leaf pieces of ten plant species common to southern Spain inoculated with conidia of two compatible isolates of D. rabiei, and formed pseudothecia with asci and viable ascospores on six of ten species and pycnidia with conidia on all plant species.     

The life cycle of <Emphasis Type="Italic">Stemphylium vesicarium</Emphasis>, the causal agent of Welsh onion leaf blight

Symptoms of Welsh onion leaf blight, caused by Stemphylium vesicarium, are divided into two types, i.e., brown oval lesions and yellow mottle lesions. Yellow mottle lesions exert considerable economic damage on Welsh onion in northern Japan. In this study, we investigated the life cycle of the pathogen in terms of seasonal fluctuation of spore dispersal and its relationship with development of disease, formation period of pseudothecia and overwintering of the pathogen based on field surveys, spore trapping and fungal isolation. Conidia were trapped throughout the cropping season except before mid June, when no ascospores were trapped. Brown oval lesions, which contained a large number of conidia, usually occurred in July followed by yellow mottle lesions with an increasing number of conidia trapped. These observations suggest that conidia released from brown oval lesions play an important role as a secondary inoculum source of the disease, leading to the development of yellow mottle lesions. Pseudothecia on leaves were first observed at the end of the cropping season or immediately after harvest (late October). The pathogen overwintered in the form of pseudothecia produced on leaves with or without symptoms. Ascospores failed to be trap in the field during the interval between before and beginning of the cropping season in April–May. However, pot experiments demonstrated that ascospores were released from leaf debris in November and rapidly increased in number after snow melt. From this circumstantial evidence, we hypothesize that ascospores are the primary inoculum source of Welsh onion leaf blight.     

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.     

Phylogenetic Analysis of Downy Mildew Pathogens of Opium Poppy and PCR-Based In Planta and Seed Detection of Peronospora arborescens

ABSTRACT Severe downy mildew diseases of opium poppy (Papaver somniferum) can be caused by Peronospora arborescens and P. cristata, but differentiating between the two pathogens is difficult because they share morphological features and a similar host range. In Spain, where severe epidemics of downy mildew of opium poppy have occurred recently, the pathogen was identified as P. arborescens on the basis of morphological traits. In this current study, sequence homology and phylogenetic analyses of the internal transcribed spacer regions (ITS) of the ribosomal DNA (rDNA) were carried out with DNA from P. arborescens and P. cristata from diverse geographic origins, which suggested that only P. arborescens occurs in cultivated Papaver somniferum in Spain. Moreover, analyses of the rDNA ITS region from 27 samples of downy-mildew-affected tissues from all opium-poppy-growing regions in Spain showed that genetic diversity exists within P. arborescens populations in Spain and that these are phylogenetically distinct from P. cristata. P. cristata instead shares a more recent, common ancestor with a range of Peronospora species that includes those found on host plants that are not members of the Papaveraceae. Species-specific primers and a PCR assay protocol were developed that differentiated P. arborescens and P. cristata and proved useful for the detection of P. arborescens in symptomatic and asymptomatic opium poppy plant parts. Use of these primers demonstrated that P. arborescens can be transmitted in seeds and that commercial seed stocks collected from crops with high incidence of the disease were frequently infected. Field experiments conducted in microplots free from P. arborescens using seed stocks harvested from infected capsules further demonstrated that transmission from seedborne P. arborescens to opium poppy plants can occur. Therefore, the specific-PCR detection protocol developed in this study can be of use for epidemiological studies and diagnosing the pathogen in commercial seed stocks; thus facilitating the sanitary control of the disease and avoidance of the pathogen distribution in seeds.     

Sporulation of Pyrenopeziza brassicae (light leaf spot) on oilseed rape (Brassica napus) leaves inoculated with ascospores or conidia at different temperatures and wetness durations

In controlled environment experiments, sporulation of Pyrenopeziza brassicae was observed on leaves of oilseed rape inoculated with ascospores or conidia at temperatures from 8 to 20°C at all leaf wetness durations from 6 to 72 h, except after 6 h leaf wetness duration at 8°C. The shortest times from inoculation to first observed sporulation ( l ₀), for both ascospore and conidial inoculum, were 11–12 days at 16°C after 48 h wetness duration. For both ascospore and conidial inoculum (48 h wetness duration), the number of conidia produced per cm² leaf area with sporulation was seven to eight times less at 20°C than at 8, 12 or 16°C. Values of Gompertz parameters c (maximum percentage leaf area with sporulation), r (maximum rate of increase in percentage leaf area with sporulation) and l ₃₇ (days from inoculation to 37% of maximum sporulation), estimated by fitting the equation to the observed data, were linearly related to values predicted by inserting temperature and wetness duration treatment values into existing equations. The observed data were fitted better by logistic equations than by Gompertz equations (which overestimated at low temperatures). For both ascospore and conidial inoculum, the latent period derived from the logistic equation (days from inoculation to 50% of maximum sporulation, l ₅₀) of P. brassicae was generally shortest at 16°C, and increased as temperature increased to 20°C or decreased to 8°C. Minimum numbers of spores needed to produce sporulation on leaves were ≈25 ascospores per leaf and ≈700 conidia per leaf, at 16°C after 48 h leaf wetness duration.     

Discovery of Didymella fabae sp. nov., the teleomorph of Ascochyta fabae, on faba bean straw

The teleomorph of Ascochyta fabae has been recorded for the first time on overwintering bean straw of Viciafaba in Cambridge. Single ascospores gave rise to typical cultures of A. fabae, the conidia of which infected faba bean plants to give ascochyta blight. Comparison with similar fungi described from Vicia spp. indicated that this is an undescribed species. The name Didymella fabae Jellis & Punith. is introduced for this teleomorph and its significance in the epidemiology of the disease is discussed.     

Phoma–Didymella complex on hybrid arctic bramble with wilting symptoms

Pycnidia containing conidia characteristic of Phoma spp. and pseudothecia containing ascospores characteristic of Didymella applanata were isolated from edges of expanding stem lesions and dead stems of wilted cultivated hybrid arctic bramble plants ( Rubus arcticus nothossp. stellarcticus ) in Sweden in 1998 and 1999. The fungi were morphologically similar when grown on culture media, but some differences in growth rate were observed. They were also similar to the reference isolates of D. applanata (anamorph Phoma argillacea ). However, they were different from an isolate of Phoma sp. (HPP 38) isolated from cultivated arctic bramble ( R. arcticus ssp. arcticus ) in Finland in 1980, and from reference isolates of Phoma glomerata isolated from other hosts. Multivariate analysis of growth rate data and conidial dimensions measured in vitro indicated that the fungi isolated from hybrid arctic bramble in Sweden were not distinguishable from D. applanata , but were clearly distinct from P. glomerata and P. exigua. Furthermore, they had identical ITS1 and ITS2 sequences, and were placed in a phylogenetic clade very closely related to the clade that contained isolates of D. applanata isolated from raspberry ( Rubus idaeus ). In contrast, isolate HPP 38 from Finland was placed in a clade with P. exigua. These data indicate that the Swedish isolates infecting arctic bramble belong to a strain of D. applanata that differs from the isolate infecting raspberry only by two common nucleotide substitutions in ITS2. Fungi of the Phoma–Didymella complex on wild and cultivated arctic bramble (a total of 291 plants showing symptoms sampled from 37 sites) were detected by a PCR-based assay and found to be common in northern Sweden, but rare, albeit widely distributed, in Finland.     

Role of Cylindrosporium-type conidia of Mycosphaerella pomi (Pass.) Lindau: cause of Brooks fruit spot of apple, as an infection source in apple orchards

Ascospores of Mycosphaerella pomi, the pathogen of Brooks fruit spot of apple, were produced in pseudothecia on previously infected and overwintered apple leaves from late April through early August in Aomori Prefecture, Japan. In June 2003, the ascospores were germinating and producing Cylindrosporium-type conidia on apple fruit and leaf surfaces in an orchard. After ascospores were sprayed on apple leaves, Cylindrosporium-type conidia developed on the leaf surfaces. Such Cylindrosporium-type conidia caused typical symptoms of Brooks fruit spot on apple trees after inoculations. These results suggested that the Cylindrosporium-type conidia also serve as an infection source, in addition to the ascospores, for Brooks fruit spot in apple orchards.     

Biological Control of Botrytis cinerea in Cyclamen with Ulocladium atrum and Gliocladium roseum Under Commercial Growing Conditions

ABSTRACT The effect of treatments with conidial suspensions of Ulocladium atrum and Gliocladium roseum on leaf rot of cyclamen caused by Botrytis cinerea was investigated under commercial greenhouse conditions. Spraying U. atrum (1 x 10(6) conidia per ml) or G. roseum (2 x 10(6) conidia per ml and 1 x 10(7) conidia per ml) at intervals of 2 to 3 weeks during the production period and spraying U. atrum (1 x 10(6) conidia per ml) at intervals of 4 to 6 weeks resulted in a significant reduction of natural infections of petioles by B. cinerea. U. atrum or G. roseum (1 x 10(7)conidia per ml) was as effective as the standard fungicide program. B. cinerea colonized senesced leaves within the plant canopy and infected adjacent petioles and leaves later. The antagonists colonized senesced leaves and reduced B. cinerea development on these leaves. Thus, the inoculum potential on petioles adjacent to necrotic leaf tissues was reduced. The fate of U. atrum conidia on surfaces of green cyclamen leaves during a 70-day period after application was studied. The number of conidia per square centimeter of leaf surface remained relatively constant during the entire experiment. Sixty percent of the conidia sampled during the experiments retained the ability to germinate. When green leaves were removed from the plants to induce senescence and subsequently were incubated in a moist chamber, U. atrum colonized the dead leaves. Senesced leaves also were colonized by other naturally occurring fungi including B. cinerea. On leaves treated with U. atrum from all sampling dates, sporulation of B. cinerea was significantly less as compared with the untreated control. Our results indicate that early applications of U. atrum before canopy closure may be sufficient to achieve commercially satisfactory control of Botrytis leaf rot in cyclamen.     

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.     

Airborne ascospores ofDidymella rabiei as a major primary inoculum for Ascochyta blight epidemics in chickpea crops in southern Spain

The incidence and severity of Ascochyta blight in potted chickpea trap plants exposed for 1-wk periods near infested chickpea debris in Córdoba, Spain, or in chickpea trap crops at least 100 m from infested chickpea debris in several locations in southern Spain were correlated with pseudothecial maturity and ascospore production ofDidymella rabiei from nearby chickpea debris. The period of ascospore availability varied from January to May and depended on rain and maturity of pseudothecia. The airborne concentration of ascospores ofD. rabiei was also monitored in 1988. Ascospores were trapped mostly from the beginning of January to late February; this period coincided with that of maturity of pseudothecia on the chickpea debris. Most ascospores were trapped on rainy days during daylight and 70% were trapped between 12.00 and 18.00 h. Autumn-winter sowings of chickpea were exposed longer to ascospore inoculum than the more traditional spring sowings because the autumn-winter sowings were exposed to the entire period of ascospore production on infested chickpea debris lying on the soil surface.     

Estimating the dynamics of airborne ascospores of Venturia inaequalis*

A system was elaborated to estimate the dynamics of primary inoculum of Venturia inaequalis in apple orchards. It separates the primary inoculum season into five periods with different risks: absent (ascospores not yet mature); potential (ascospores mature but not yet ready to be discharged); actual (ascospores can be discharged when favourable conditions occur); present (ascospores are airborne); exhausted (all ascospores have been ejected). These periods were determined by two mathematical models, which use meteorological parameters as driving variables. The first model estimates the development stage of the overwintering pseudothecia and then determines when the first pseudothecia contain pigmented and mature ascospores. A threshold of mature ascospores inside pseudothecia defines when the ascospores become ready for discharge. The second model estimates the proportion of the season's ascospores that are airborne on each discharging event, using temperature and leaf wetness, expressed as the degrees accumulated daily in the hours when leaves are wet. Estimates of absent and potential risk were verified by collecting data on the first ascospore discharge in the period 1991/1998 at Bologna and Modena (northern Italy), and they were always found to be accurate. To verify the estimates of actual, present and exhausted risk, the model outputs were compared with data collected by spore samplers at Modena and Bologna in 1997 and 1998: they were sufficiently accurate because the greatest part of the records from the spore sampler fell inside the confidence limits of the model.     

Role of oospores as primary inoculum for epidemics of downy mildew caused by Peronospora arborescens in opium poppy crops in Spain

This study investigated the role of Peronospora arborescens oospores as primary inoculum for downy mildew of opium poppy and infection types that they may give rise to in Spain using an integrative experimental approach that combined pathogenicity tests in growth chambers and field microplots, together with molecular detection of P. arborescens infection by specific nested-PCR assays. The results demonstrated that oospores in infested soil or leaf debris were effective inoculum for ingress of the pathogen through underground plant tissues early in poppy seedling growth. This gave rise to systemic infections that reproduced the stunting, chlorotic syndrome frequently observed in affected plants in commercial fields. Additionally, infection of underground tissues of older plants by oospore inoculum could remain asymptomatic. Results also suggested that sporangia formed on infected plants are effective in producing secondary local infections that later may become systemic and either symptomatic or asymptomatic. Finally, and more importantly, those delayed symptomatic or asymptomatic systemic infections, as well as secondary local infections of capsules, can give rise to infected seeds. This research on the biology of P. arborescens on poppy plants and epidemiology of downy mildew may help to develop knowledge-based disease-management strategies of use in the protection of yields of opium poppy crops in Spain and elsewhere.     

Environmental Factors Affecting Pseudothecial Development and Ascospore Production of Mycosphaerella citri, the Cause of Citrus Greasy Spot

ABSTRACT Mycosphaerella citri, the cause of citrus greasy spot, produces pseudothecia and ascospores in decomposing leaf litter on the grove floor. In laboratory studies, the effect of wetting and drying and temperature on the formation, maturation, and production of pseudothecia and ascospores was evaluated on mature, detached grapefruit leaves. Production of pseudothecia was most rapid when leaves were soaked five times per week for 2 h per day, but pseudothecial density and total ascospore production were greatest when leaves were soaked three times per week for 2 h per day. In duration of wetting studies, 3 h per day, 3 days per week brought about the most rapid production, but 10 to 30 min per day resulted in production of the most pseudothecia and ascospores. Pseudothecia and ascospore production were greatest at 28 degrees C and declined rapidly at lower and higher temperatures. Maturation of pseudothecia was slow at 20 and 24 degrees C, but production was high at 24 degrees C; at 32 degrees C, pseudothecia matured rapidly, but degenerated quickly. No mature pseudothecia were produced on leaves maintained continuously under wet conditions. In field studies, leaves were placed on the grove floor monthly from April 2000 to September 2001. Pseudothecia production was rapid during the summer rainy season from June to September. Pseudothecia produced on leaves placed in the grove from October to May developed and matured more slowly but were produced in much larger numbers than in summer. The number of days to first pseudothecial initials, 50% maturation, first discharge of ascospores, leaf decomposition, as well as pseudothecial density and incidence, were negatively related to average temperature. Total ascospore production was unrelated to temperature.     

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.     

The effect of winter weather conditions on the ability of pseudothecia of Leptosphaeria maculans and L. biglobosa to release ascospores

Leptosphaeria maculans and L. biglobosa are damaging pathogens of oilseed rape. The infection of plants occurs predominantly in early autumn or spring by spores produced in pseudothecia. The aim of this study was to investigate whether pseudothecia formed in the autumn are still viable in the spring and to what extend they are destroyed by winter frosts. The studies presented here demonstrated that winter frosts can render pseudothecia unable to release spores. Nevertheless, ascospores present in pseudothecia unable to discharge ascospores, were fully capable of germination, regardless of the incubation temperature. No significant differences were found between the studied Leptosphaeria species in their response to frost. A multiple regression equation has been elaborated to forecast the ability of pseudothecia to release ascospores, based on winter temperatures. Considerable correlation was found between the ascospore release in the autumn and the ability of pseudothecia to release ascospores over the winter period and the subsequent symptoms of stem canker before harvest. We have demonstrated that the potential and the survival of inoculum can have a large impact on the success of the pathogen. This may be particularly important in the light of forecasted climate change. Higher winter temperatures may increase the ability of pseudothecia to release ascospores and the discharge of ascospores of L. maculans and L. biglobosa into the air, and cause early plant infections. This in turn will increase the number of infected plants, the disease incidence at harvest, and reduce the yield of oilseed rape.