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1.
Spores of many fungal pathogens are dispersed by wind. Detection of these airborne inocula is important in forecasting both the onset and the risk of epiphytotics. Species-specific primers targeted at the internal transcribed spacer (ITS) region ofLeptosphaeria maculans andL. biglobosa — the causal organisms of phoma stem canker and stem lesions ofBrassica spp., including oilseed rape — were used to detect DNA extracted from particles deposited on tapes obtained from a spore trap operated in Rarwino (northwest Poland) from September to November in 2004 and 2006. The quantities of DNA assessed by traditional end-point PCR and quantitative real-time PCR were compared to microscopic counts of airborne ascospores. Results of this study showed that fluctuations in timing of ascospore release corresponded to the dynamics of combined concentrations of DNA fromL. maculans andL. biglobosa, with significant positive correlations between ascospore number and DNA yield. Thus the utilization of PCR-based molecular diagnostic techniques enabled the detection, identification, and accurate quantification of airborne inoculum at the species level. Moreover, real-time PCR was more sensitive than traditional PCR, especially in years with low ascospore numbers.  相似文献   

2.
When airborne propagules of Leptosphaeria maculans and L. biglobosa were collected in Poland at three ecologically different sites from 1 September to 30 November in 2004 to 2008, using a Hirst-type seven-day volumetric spore trap, there were fluctuations in timing of ascospore release and diverse ratios between airborne propagules of both species depending on season, field location and weather conditions. The detection was done using the microscope as well as quantitative PCR with species-specific primers targeted against fragments of β-tubulin genes and quantified with a dual-labelled fluorescent probe approach. This detection chemistry is described for the first time for L. maculans and L. biglobosa. Its advantage over the previous ITS-based SYBR-Green chemistry resides in improved sensitivity and the virtual absence of false positives in the detection of these fungi. There were significant, positive correlations between data obtained using visual assessment of ascospore numbers and DNA concentrations that were measured by qPCR. Climatic differences between the oilseed rape growing regions could have significantly affected the biological processes of pseudothecial maturation and ascospore development of the pathogens. The data suggest that regular rain events of intermediate intensity recorded in the Maritime region favoured the maturation of the pathogen more than the drier weather recorded in the Silesia or Pomerania regions. It was observed that the number of rainy days was of greater importance than the cumulative rainfall to obtain the generative sporulation of the pathogen. Accurate detection of airborne inoculum of pathogenic Leptosphaeria spp. facilitates improved targeting of disease management decisions for oilseed rape protection against phoma stem canker and stem necrosis diseases.  相似文献   

3.
Banana leaves showing different levels of black Sigatoka disease were collected from an unsprayed plantation in Costa Rica during two separate periods representing the wet to dry season transition (October 1993 – February 1994) and the dry to wet season transition (April – September 1995). Laboratory studies were used to investigate the relationship between the release of Mycosphaerella fijiensis ascospores and the amount of inoculum on banana plants showing different levels of infection, as assessed by leaf necrotic area. The number of perithecia present in the necrotic area was used as an indication of potential ascospore loads and was investigated as a series of regression equations. A series of rewetting and incubation regimes was used to investigate spore release under field conditions (21°C and 100% relative humidity in the early morning and 28°C, 60% relative humidity on days when it rained in mid-afternoon). Results suggest that rainfall, combined with a high temperature, may lead to peaks of ascospore release but without necessarily increasing overall numbers released over periods of up to 4 days and that a high level of spore release was less sensitive to changes in temperature once it had been initiated. The exact role of temperature in spore release is still unclear, however, as leaf samples kept at atypically low temperatures also released non-germinating ascospores. An average of 4.5 ascospores was released per perithecium. This does not resolve ambiguities in the literature regarding the number of ascospores present in each perithecium. A linear model relating the average ascospore numbers and necrotic area, using quick estimates of the amounts of necrotic area on the leaves of a random sample of plants across a plantation, is proposed, to give an indication of the relative amount of airborne inoculum potentially available between different plantations.  相似文献   

4.
Diurnal patterns of spore release have been observed in a number of fungal pathogens that undergo wind-assisted dispersal. The mechanisms that drive these patterns, while not well understood, are thought to relate to the ability of dispersing spores to survive their journey and infect new hosts. In this paper, we characterise the diurnal pattern of ascospore release by a Western Australian population of Leptosphaeria maculans. Although L. maculans has been previously shown to exhibit diurnal patterns of ascospore release, these patterns appear to vary from region to region. In order to characterise the pattern of release in the Mediterranean climate of Western Australia, we analysed historical data describing the bi-hourly count of airborne ascospores at Mt Barker, Western Australia. Results of this analysis showed diurnal patterns that differ from those previously observed in other countries, with ascospore release in our study most likely to occur in the afternoon. Furthermore, we found that the time of peak release can shift from month to month within any one season, and from year to year. In explaining the hourly pattern of spore release over an entire season, time since rainfall, time since last release, temperature, hour and month were all shown to be significant variables.  相似文献   

5.
Data from a controlled environment experiment investigating effects of temperature on maturation of Leptosphaeria maculans pseudothecia were used to derive equations describing the times until 30% or 50% of pseudothecia were mature as a function of temperature. A wetness sensor was developed to estimate the oilseed rape debris wetness and operated with debris exposed in natural conditions in 2000 and 2001. The maturation of L. maculans pseudothecia on debris and concentrations of airborne L. maculans ascospores were observed from 1999 to 2004. There were considerable differences between years, with the first mature pseudothecia observed in September in most years. There were linear relationships between the first date when 10% of maximum ascospore release was observed and the dates when 30% or 50% of pseudothecia were mature. By summing the daily temperature‐dependent rate of pseudothecial maturation for days after 1 August with rainfall >0.5 mm, the dates when 30% or 50% of pseudothecia were mature were predicted. There was good agreement between predicted and observed dates when 30% or 50% of pseudothecia were mature. These equations for predicting the timing of L. maculans ascospore release could be incorporated into schemes for forecasting, in autumn, the severity of phoma stem canker epidemics in the following spring/summer in the UK.  相似文献   

6.
Abstract The influence of air temperature on the release pattern of Venturia inaequalis ascospores was studied by volumetric spore samplers in two spore sampling periods. In the first period (1991–1996; Passo Segni, Ferrara), 15 ascospore dispersal events were considered occurring in daylight, with high spore counts (168–5892 ascospores per m3 air per event), at an average temperature between 8.4 and 20.3°C. Both the length of the ascospore release period and distribution of airborne spores over time were significantly influenced by temperature. A logistic regression model was used to fit the proportion of ascospores trapped from the orchard air as a function of time after the beginning of the discharge event and air temperature. The accuracy of this equation was tested against data collected in the second spore sampling period (1997–2000; Sala Bolognese, Bologna, and Castelfranco, Modena); 16 dispersal events were considered, triggered by rainfall that occurred both in the dark and in daylight, with low to high spore counts (29–458 ascospores per m3 air per event), at an average temperature between 2.8 and 14.3°C. There was a general agreement between the proportion of ascospores trapped from the orchard air during these events and that estimated by using the logistic equation – in most cases, actual and estimated values showed a high coincidence. Statistical comparison showed a significant correlation (r=0.93, P < 0.01) between observed and estimated data.  相似文献   

7.
The dynamics of airborne ascospores and disease progress of red leaf blotch (RLB) of almond, caused by Polystigma amygdalinum, and their correlations with weather variables were studied from 2019 to 2021 in two almond orchards located in Lleida, NE Spain. Airborne ascospores were detected and quantified by real-time qPCR using species-specific primers for P. amygdalinum. Ascospores were detected mainly from April to June, with a high variability between the yearly cumulative concentrations. Positive significant correlations were found between the weekly proportion of airborne ascospores and the number of wet and mild days—either combined or separated— accumulated rainfall, number of rainy days, accumulated low temperatures on wet days, and mean and maximum relative humidity. In contrast, several thermal variables (maximum temperature, VPD, and number of warm days) were negatively correlated with ascospore catches. Positive significant correlations were found between the cumulative proportion of ascospores and RLB incidence and severity. Weekly variations in RLB incidence and severity showed significant positive correlations with the number of warm days while negative with the number of mild days. Severity was also positively correlated with several thermal variables (mean, maximum, and minimum temperature, and VPD), and negatively correlated with the number of cold days and wet and mild days. Stronger correlations were generally found with ascospore catches or disease progress when using concurrent weekly weather data. Gompertz, monomolecular, and logistic growth models were evaluated to describe RLB disease progress.  相似文献   

8.
A selection of fungicides, herbicides and surfactants and urea were tested for their effect on the production of pseudothecia and ascospore release of Leptosphaeria maculans present on oil-seed rape straw and turnip seed crop straw. The fungicides ethyl mercury phosphate, triarimol, fenarimol, carbendazim, tridemorph and benomyl, each at 1 g/litre, the herbicides dinoseb and diquat, each at 10 g/litre the surfactants Bradasol, Cetrimide, Deciquam 222, Hyamine 1622 and Maxonol N, each at 50 g product/litre, and urea at 150 g/litre, applied to straw before pseudothecia had formed were more than 90% effective in preventing further development. These chemicals were also effective in preventing further ascospore production when applied to straw bearing mature pseudothecia but only dinoseb and urea prevented the release of mature ascospores. The results suggest that it may be possible to break the life cycle of L. maculans by chemical treatment and thereby obviate the need for subsequent control measures.  相似文献   

9.
Airborne ascospores have been reported to be allergenic or plant pathogens, and their presence has traditionally been associated with rainfall events. The aim of the present study was to analyze the presence of airborne ascospores in relation to weather parameters in a town in SW Spain. A seven-day recording spore trap (Burkard) was used to sample the air over 2 years at 15 m above ground level on the terrace roof of the hospital in Mérida (SW Spain). Fungal spores were identified and counted by means of two longitudinal scans over the slides under ×1000 microscopy. A correlation analysis was made of the daily meteorological data and the airborne ascospore concentrations, and t-tests were used to compare data between the 2 years. Nineteen ascospore types were defined, including one-cell ascospores (Chaetomium, Diatrype, Helvella, Xylaria), tow-cell ascospores (Diaporthe, Mycosphaerella, Nectria, Venturia), transversally septate ascospores (Melanomma, Leptosphaeria, Paraphaeosphaeria, Sporormiella, Massaria), transversally and longitudinally septate ascospores (Pleospora), and ascospores within asci (Sordaria). Leptosphaeria consisted of a group of four types described according to the number of cells, hyaline grade, wall thickness, and ornamentation, and other ascospores comprised one last additional type. The average airborne ascospore concentration was 153 ascospores/m3. One-third each of this total were from the Leptosphaeria group, with an average 54 ascospores/m3, and the two-cell ascospores or Venturia-like group (Diaporthe, Mycosphaerella, Nectria, Venturia) with 51 ascospores/m3 on average. In third position was Pleospora with 27 ascospores/m3 on average. The month with highest concentration was September, with 238 ascospores/m3, and the lowest March, with 56 ascospores/m3. By seasons, autumn had the highest concentrations, followed by winter, spring, and summer. The maximum daily concentration reached was 3,371 ascospores/m3. Daily rainfall was significantly correlated with the ascospore types Diatrype, Mycosphaerella, Nectria, two subtypes of Leptosphaeria, and Pleospora. Relative humidity was positively correlated with those ascospore types and also with Diaporthe and Paraphaeosphaeria, and negatively with Chaetomium and Melanomma. The concentration was higher on rainy days than on days without rain for Pleospora, Leptosphaeria (3 subtypes), Diatrype, Diaporthe, Nectria, Mycosphaerella, and Paraphaeosphaeria. The daily temperatures were in general correlated with the same types as the relative humidity, but with the opposite sign. For the monthly data, there were no statistically significant differences between the 2 years studied.  相似文献   

10.
Mycelium of Leptosphaeria maculans survived on oilseed rape stem base debris buried in sand for 2,4, 6, 8,10 or 12 months and produced pseudothecia after subsequent exposure on the surface of the ground under natural conditions for 2–4 months, but did not survive on upper stem debris buried for 2 months. Only A‐group L. maculans ascospores were produced on the stem base debris which had been buried; no B‐group ascospores were produced. Mycelium of L. maculans survived on both stem base and upper stem debris exposed on the sand surface for 2, 4, 6, 8, 10 or 12 months and pseudothecia with viable ascospores were observed at the time of sampling. Both A‐group L. maculans (predominant on stem bases) and B‐group L. maculans (predominant on upper stems) ascospores were produced on unburied stem base and upper stem debris. Thus B‐group L. maculans survived longer on unburied debris than on buried debris. A‐group ascospores which were exposed in dry air in darkness at 5–20°C survived longer than B‐group ascospores; 10–37% of A‐group ascospores, compared with 2–31% of B‐group ascospores, survived after 35 days.  相似文献   

11.
A 6-year study was carried out in an apple-growing region of North Italy by trapping airborne ascospores of Venturia inaequalis with a volumetric spore trap operated continuously during the ascospore season, with the aim of better defining the weather conditions that allow ascospores both to discharge and to disperse into the orchard air. A total of more than 60 ascospore trapping events occurred. Rain events were the only occurrences allowing ascospores to become airborne (a rain event is a period with measurable rainfall ≥0.2 mm/h – lasting one to several hours, uninterrupted or interrupted by a maximum of two dry hours); on the contrary, dew was always insufficient to allow ascospores to disperse into the air at a measurable rate, in the absence of rain. In some cases, rain events did not cause ascospore dispersal; this occurred when: (i) rain fell within 4–5 h after the beginning of a previous ascospore trapping; (ii) rain fell at night but the leaf litter dried rapidly; (iii) nightly rainfalls were followed by heavy dew deposition that persisted some hours after sunrise. Daytime rain events caused the instantaneous discharge and dispersal of mature ascospores so that they became airborne immediately; for night-time rainfall there was a delay, so that ascospores became airborne during the first 2 h after sunrise. This delay did not always occur, and consequently the ascospore trapping began in the dark, when: (i) the cumulative proportion of ascospores already trapped was greater than 80% of the total season's ascospores; (ii) more than one-third of the total season's ascospores was mature inside pseudothecia and ready to be discharged.  相似文献   

12.
The ability of Sphaerotheca mors-uvae to perennate as cleistocarps, and as mycelium in buds was examined during the winters of 1965-6, 1966-7 and 1967-8 in relation to its two principal hosts, gooseberry and black currant. Cleistocarps on black currant leaves were examined from August 1965 to April 1966 and from July 1966 to March 1967. In 1965 cleistocarps were first observed on the leaves on 5 August; in 1966 on 11 July. These continued to develop through August and September and by October approximately 70% contained well-defined ascospores. The ascospore content remained generally at this level until February 1966 and November 1966; then the numbers of cleistocarps with ascospores fell and by April 1966 and March 1967 few such cleistocarps remained. From 21 March 1966 and 15 February 1967, but not otherwise, discharge of ascospores from the overwintered cleistocarps was readily obtained in laboratory tests. The viability and infectivity of the ascospores was demonstrated by allowing them to discharge on to leaf discs of black currant in the laboratory and also on to leaf discs and plants in the field. Sporulating colonies of S. mors-uvae developed within 8 days. Cleistocarps from shoots of black currant were examined from 4 August 1966 to 9 March 1967, and from 27 July 1967 to 1 January 1968. They developed in a similar manner to those on black currant leaves and by September in both 1966 and 1967 over 60% contained ascospores. This level was not maintained; the number of cleistocarps with ascospores fell gradually and by 8 December 1966 and 1 January 1968 few remained. Only in one laboratory test (21 November 1967) were ascospores discharged from a sample of these cleistocarps. Cleistocarps from shoots of gooseberry were examined from July 1966 to March 1967, and from August 1967 to January 1968. The pattern of ascospore development and subsequent decline in number of cleistocarps with ascospores was similar to that observed for black currant shoots. No discharge of ascospores could be demonstrated in laboratory tests. Evidence that S. mors-uvae perennates in buds of gooseberry was obtained by dissecting buds and by inducing buds on surface-sterilized shoots to burst under conditions which precluded chance infection. Field observations also suggested that bud infection occurred on gooseberry. Similar experiments failed to demonstrate the fungus in buds of black currant, and there was no indication of bud infection of this host in the field.  相似文献   

13.
In 1988 and 1989 a plot containing 668 sunflower plants (cv. Sunbred 246) was inoculated with similar amounts of sclerotia of Sclerotinia sclerotiorum in February or March and disease development was monitored on each plant during the summer. The concentration of airborne ascospores in the plots was measured and was found to correspond to the number of apothecia. In both years ascospore production started early in June and in 1988 reached a peak in mid-July and lasted until the beginning of August. However, in 1989 ascospore production only lasted for about two weeks and airborne concentrations were small. Ascospores were released predominantly during the day, mostly around 1200 BST. Disease symptoms were not observed until July, between 25 and 40 days after ascospores were first found. Disease incidence increased roughly linearly with time until mid-August when the rate of infection declined. In 1988 the number of plants per week with new symptoms was roughly proportional to the average ascospore concentration measured 5 weeks previously. The observations suggest that severity of disease may be related to ascospore concentration during the infection period.  相似文献   

14.
Leptosphaeria maculans and L. biglobosa are closely related sibling fungal pathogens that cause phoma leaf spotting, stem canker (blackleg) and stem necrosis of oilseed rape (Brassica napus). The disease is distributed worldwide, and it is one of the main causes of considerable decrease in seed yield and quality. Information about the time of ascospore release at a particular location provides important data for decision making in plant protection, thereby enabling fungicides to be used only when necessary and at optimal times and doses. Although the pathogens have been studied very extensively, the effect of climate change on the frequencies and distributions of their aerially dispersed primary inoculum has not been reported to date. We have collected a large dataset of spore counts from Poznan, located in central-west part of Poland, and studied the relationships between climate and the daily concentrations of airborne propagules over a period of 17 years (1998–2014). The average air temperature and precipitation for the time of development of pseudothecia and ascospore release (July–November), increased during the years under study at the rates of 0.1 °C and 6.3 mm per year. The day of the year (DOY) for the first detection of spores, as well as the date with maximum of spores, shifted from 270 to 248 DOY, and from 315 to 265 DOY, respectively. The acceleration of the former parameter by 22 days and the latter by 50 days has great influence on the severity of stem canker of oilseed rape.  相似文献   

15.
16.
The populations of Monosporascuscannonballus ascospores in the soils of 14 muskmelon fields throughout muskmelon production areas of Comunidad Valenciana (eastern Spain) were quantified from 2002 to 2003. The fields were surveyed in July, at the end of the cropping season, when plants approached maturity and symptoms of vine decline in the canopy appeared as patches of wilted or dead plants. Ascospores were recovered from all muskmelon fields surveyed. The average soil populations ranged from 1.54 to 6.33 per g of soil and the number of ascospores within a soil sample ranged from 0.50 to 16.20 ascospores/g of soil. This is the first detailed report on ascospore populations of M. cannonballus in muskmelon fields from a cucurbit‐growing area in Spain and the Mediterranean basin.  相似文献   

17.
Upon cultivation of the yeast Dipodascopsis tothii in its sexual stage, small ascospores are released individually from the ascus tip, which then assemble in sheathed cluster balls. In contrast to Dipodascopsis uninucleata, this yeast produced smooth bean shaped ascospores with sheath-like appendages that assemble in a disordered sheathed ball of ascospores outside the ascus. Strikingly, upon release, the ascus tip contained 3-hydroxy oxylipins, while the released ascospore clusters contained little or no 3-hydroxy oxylipins as indicated by immunofluorescence microscopy. In D. uninucleata, these oxylipins are concentrated on the spore surface and interspore matrix, but not on the ascus tip.  相似文献   

18.
The effects of air humidity and temperature on the ascospore discharge of Graphostroma platystoma were experimentally investigated. The ascospores were not discharged from the stromata in air at 100% relative humidity (RH). However, they were discharged from the wetted stromata at 3°, 10°, and 24°C under 100% RH or nearly so. The amount of the discharged ascospore was large at 24°C, medium at 10°C, and small at 3°C. The ascospores in the rainwater that washed down the stromata were counted after rainfall in the field. The discharge was observed from September to the following May.  相似文献   

19.
Quantitative resistance against Leptosphaeria maculans in Brassica napus is difficult to assess in young plants due to the long period of symptomless growth of the pathogen from the appearance of leaf lesions to the appearance of canker symptoms on the stem. By using doubled haploid (DH) lines A30 (susceptible) and C119 (with quantitative resistance), quantitative resistance against L. maculans was assessed in young plants in controlled environments at two stages: stage 1, growth of the pathogen along leaf veins/petioles towards the stem by leaf lamina inoculation; stage 2, growth in stem tissues to produce stem canker symptoms by leaf petiole inoculation. Two types of inoculum (ascospores; conidia) and three assessment methods (extent of visible necrosis; symptomless pathogen growth visualised using the GFP reporter gene; amount of pathogen DNA quantified by PCR) were used. In stage 1 assessments, significant differences were observed between lines A30 and C119 in area of leaf lesions, distance grown along veins/petioles assessed by visible necrosis or by viewing GFP and amount of L. maculans DNA in leaf petioles. In stage 2 assessments, significant differences were observed between lines A30 and C119 in severity of stem canker and amount of L. maculans DNA in stem tissues. GFP-labelled L. maculans spread more quickly from the stem cortex to the stem pith in A30 than in C119. Stem canker symptoms were produced more rapidly by using ascospore inoculum than by using conidial inoculum. These results suggest that quantitative resistance against L. maculans in B. napus can be assessed in young plants in controlled conditions. Development of methods to phenotype quantitative resistance against plant pathogens in young plants in controlled environments will help identification of stable quantitative resistance for control of crop diseases.  相似文献   

20.
Fusarium graminearum is a predominant component of the Fusarium head blight (FHB) complex of small grain cereals. Ascosporic infection plays a relevant role in the spread of the disease. A 3-year study was conducted on ascospore discharge. To separate the effect of weather on discharge from the effect of weather on the production and maturation of ascospores in perithecia, discharge was quantified with a volumetric spore sampler placed near maize stalk residues bearing perithecia with mature ascospores; the residues therefore served as a continuous source of ascospores. Ascospores were discharged from perithecia on 70% of 154 days. Rain (R) and vapor pressure deficit (VPD) were the variables that most affected ascospore discharge, with 84% of total discharges occurring on days with R≥0.2 mm or VPD≤11 hPa, and with 70% of total ascospore discharge peaks (≥ 30 ascospores/m3 air per day) occurring on days with R≥0.2 mm and VPD≤6.35 hPa. An ROC analysis using these criteria for R and VPD provided True Positive Proportion (TPP) = 0.84 and True Negative Proportion (TNP) = 0.63 for occurrence of ascospore discharge, and TPP = 0.70 and TNP = 0.89 for occurrence of peaks. Globally, 68 ascospores (2.5% of the total ascospores sampled) were trapped on the 17 days when no ascospores were erroneously predicted. When a discharge occurred, the numbers of F. graminearum ascospores sampled were predicted by a multiple regression model with R2 = 0.68. This model, which includes average and maximum temperature and VPD as predicting variables, slightly underestimated the real data and especially ascospore peaks. Numbers of ascospores in peaks were best predicted by wetness duration of the previous day, minimum temperature, and VPD, with R2 = 0.71. These results will help refine the epidemiological models used as decision aids in FHB management programs.  相似文献   

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