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1.
Effects of pretreatment of Brassica napus leaves with ascospores of Leptosphaeria biglobosa or chemical defence activators [acibenzolar- S -methyl (ASM) or menadione sodium bisulphite (MSB)] on infection by ascospores of Leptosphaeria maculans (phoma stem canker) and development of disease were studied in controlled-environment (phoma leaf spot) and field (phoma leaf spot and stem canker) experiments. In controlled-environment experiments, pretreatment of oilseed rape leaves (cv. Madrigal) with L. biglobosa , ASM or MSB delayed the appearance of L. maculans phoma leaf spot lesions. These pretreatments also decreased the phoma leaf spot lesion area in both pretreated leaves (local effect) and untreated leaves (systemic effect). In winter oilseed rape field experiments in the 2002/03 and 2003/04 growing seasons, pretreatment with L. biglobosa or ASM in October/November decreased not only the number of phoma leaf spot lesions per leaf caused by L. maculans in autumn/winter, but also the severity of phoma stem canker in the subsequent spring/summer. Effects were greater in 2002/03 (when natural L. maculans ascospore release began in September 2002) than in 2003/04 (when ascospore release began in December following a period of dry weather in August/September 2003). These results suggest that pretreatment with biological or chemical defence activators can induce local and systemic resistance to L. maculans , with both short-term effects on the development of phoma leaf spotting and long-term effects on the development of stem canker 8 months later.  相似文献   

2.
The fungus Leptosphaeria maculans causes blackleg (phoma stem canker), one of the most serious diseases of oilseed rape. The role of pycnidiospores produced during asexual reproduction is poorly documented and limits the understanding of the pathogen's population dynamics. The objectives of this study were to assess rain-splash dispersal of pycnidiospores of L. maculans from phoma leaf spots, and transmission of the disease from oilseed rape stubble carrying pycnidia. The work was conducted in still air with either a drop generator or a rain simulator. The impact of simulated incident drops on phoma leaf spots resulted in the dispersal of L. maculans pycnidiospores within splash droplets. Ninety per cent of the spores were collected within 14 cm of the source and a few were regularly observed up to 40 cm. Pycnidiospores produced on oilseed rape stubble and dispersed by simulated rain infected oilseed rape trap plants in a spatial pattern that matched the spatial dispersal of the pycnidiospores. In the field, rain-splash dispersal of pycnidiospores could increase the pathogen population and may enhance sexual reproduction by facilitating the mating of initially spatially separated isolates of opposite mating type.  相似文献   

3.
In controlled environment experiments to study early development of light leaf spot, lesions developed with leaf wetness durations of 16 to 48 h after inoculation of oilseed rape with conidial suspensions of Pyrenopeziza brassicae at 12 or 18°C, but not with leaf wetness durations of 0 to 13h. The incubation period was 21 to 22 days at 12°C and 14 to 18 days at 18°C for leaf wetness durations of 16 to 48 h. The latent period was 21 to 23 days at 12°C and 18 to 19 days at 18°C, and the total number of lesions increased with increasing leaf wetness duration at both temperatures. In field experiments, light leaf spot always developed on oilseed rape with a leaf wetness duration of 48 h after inoculation in both 1990/1991 and 1991/1992, but the percentage leaf area affected was less on plants placed in an oilseed rape crop than on those placed in a glasshouse. Plants moved to an oilseed rape crop immediately after inoculation nearly always developed light leaf spot symptoms when they were inoculated between 19 October 1990 and 1 March 1991 or between 27 September 1991 and 14 February 1992, but plants inoculated between 31 August and 16 October 1990 or on 20 September 1991, when estimated leaf wetness duration was less than 16 h for several days after they were placed in crops, did not develop symptoms. The latent period of light leaf spot on plants transferred to the oilseed rape crop was 15 to 40 days, and there was an approximately linear relationship between 1 (latent period) and mean temperature during this period. The accumulated temperature during the latent period ranged from c. 150 to 250 day-degrees. The severity of lesions on these plants increased with increasing temperature from 5 to 15°C.  相似文献   

4.
Phoma stem canker (blackleg), caused by Leptosphaeria maculans , is an important disease on oilseed rape (canola, rapeseed, Brassica napus , Brassica juncea , Brassica rapa ) causing seedling death, lodging or early senescence in Australia, Canada and Europe, but not in China. The two forms of L. maculans (A group and B group) that occur on oilseed rape are now considered to be separate species. The epidemiology and severity of phoma stem canker differs between continents due to differences in the pathogen population structure, oilseed rape species and cultivars grown, climate and agricultural practices. Epidemics are most severe in Australia, where only the A group occurs, and can be damaging in Canada and western Europe, where both A and B groups occur, although their proportions vary within regions and throughout the year. Epidemics are slight in China, where the A group has not been found. Dry climates (Australia, western Canada) lengthen the persistence of infected debris and may synchronize the release of airborne ascospores (after rain) with seedling emergence. L. maculans spreads from cotyledon and leaf infections down petioles to reach the stem, with infections on cotyledons and leaves early in the season producing the most damaging stem cankers at the stem base (crown). Development of both crown cankers and phoma stem lesions higher up stems is most rapid in regions with high temperatures from flowering to harvest, such as Australia and Canada. Breeding for resistance (genetic, disease escape or tolerance), stubble management, crop rotation and fungicide seed treatments are important strategies for control of phoma stem canker in all areas. Fungicide spray treatments are justified only in regions such as western Europe where high yields are obtained, and accurate forecasts of epidemic severity are needed to optimize their use.  相似文献   

5.
Field experiments in Europe have shown that Chinese cultivars of winter oilseed rape ( Brassica napus ) are very susceptible to the pathogen Leptosphaeria maculans (cause of phoma stem canker). Climatic and agronomic conditions in China are suitable for L. maculans since the closely related but less damaging pathogen L. biglobosa occurs on the winter and spring oilseed rape crops there. Major gene resistance to L. maculans is not durable; when introduced into commercial oilseed rape cultivars it is rapidly rendered ineffective by changes in the pathogen population. The threat to Chinese oilseed rape production from L. maculans is illustrated by the way in which L. maculans has spread into other areas of the world where previously only L. biglobosa was present, such as Canada and Poland. Models were developed to describe the spread (in space and time) of L. maculans across Alberta province, Canada, based on survey data collected over a 15-year period. These models were used to estimate the potential spread of L. maculans across the Yangtze river oilseed rape growing areas of China and its associated costs. Short-term strategies to prevent occurrence of severe phoma stem canker epidemics in China include training of extension workers to recognise symptoms of the disease and use of PCR-based diagnostics to detect the pathogen on imported seed. Long-term strategies include the introduction of durable resistance to L. maculans into Chinese oilseed rape cultivars as a component of an integrated disease management programme. The costs of such strategies in relation to costs of a phoma stem canker epidemic are discussed.  相似文献   

6.
Quantitative resistance to Leptosphaeria maculans in Brassica napus was investigated in field and controlled environments using cultivars Darmor (with quantitative resistance) and Eurol (without quantitative resistance). In field experiments, numbers of phoma leaf spot lesions in autumn/winter and severity of stem canker the following summer were assessed in three growing seasons. There were no differences between Darmor and Eurol in number of leaf lesions in autumn/winter. However, stem cankers were less severe on Darmor than Eurol at harvest the following summer. In controlled-environment experiments, development of leaf lesions at different temperatures (5–25°C) and wetness durations (12–72 h) was investigated using ascospore inoculum; symptomless growth of L. maculans along leaf petioles towards the stem was quantified using quantitative PCR and visualized using GFP-expressing L. maculans ; growth of L. maculans within stem tissues was investigated using GFP-expressing L. maculans . There were more leaf lesions on Darmor than Eurol, although there was no difference between Darmor and Eurol in L. maculans incubation period. There were no differences between Darmor and Eurol in either distance grown by L. maculans along leaf petioles towards the stem or quantity of L. maculans DNA in leaf petioles, but L. maculans colonized stem tissues less extensively on Darmor than Eurol. It was concluded that quantitative resistance to L. maculans operates during colonization of B. napus stems by the pathogen.  相似文献   

7.
Experiments to investigate the factors affecting the incubation period of dark leaf and pod spot (Alternaria brassicae) on leaves and pods of oilseed rape (Brassica napus) were done in controlled environment (constant temperatures) and glasshouse conditions (fluctuating temperatures). The length of the incubation period of dark leaf and pod spot decreased as infection and incubation temperatures increased from 6 to 20 °C. The incubation period decreased as wetness period increased from 2 to 12 h, as inoculum concentration increased from 80 to 2 × 103 spores ml–1 and as leaf age increased from 4 to 10 days. Asymptotes of leaf age and inoculum concentration, above which the length of the incubation period did not decrease, were 10 days and 2 × 103 spores ml–1, respectively. The shortest and longest incubation periods were 1 and 11 days. The mechanism by which the infection conditions influenced the incubation period of dark leaf and pod spot on oilseed rape seemed to be linked to lesion density. Usually, the length of the incubation period decreased greatly with increasing lesion density.  相似文献   

8.
On oilseed rape, 207 leaf lesions attributed to Leptosphaeria maculans were classified as typical or atypical. Starch gel electrophoresis of glucose phosphate isomerase (GPI) performed on extracts of 229 leaf lesions comprising the 207 with L. maculans symptoms and 22 with Pseudocercosporella capsellae symptoms, yielded four different electrophoretic patterns of alloenzymes designated ET1 to ET4. In addition to ET1 and ET2, characteristic respectively of A- (highly virulent) and B- (weakly virulent) group isolates of L. maculans , the previously undescribed ET3 allozyme was recovered from a few typical and atypical L. maculans leaf lesions. The fastest ET4 allozyme was specific to P. capsellae . All but two typical leaf lesions produced the ET1 allozyme, whereas atypical lesions produced one of the three L. maculans allozymes. Occasionally a mixture of two allozymes was recovered from a same-leaf lesion. GPI electrophoresis performed directly on leaf lesions proved a useful and reliable method to identify L. maculans , and to differentiate between L. maculans and P. capsellae . This method of discrimination enabled deductions, from 377 leaf lesions analysed, about the structure of L. maculans populations on different oilseed rape varieties.  相似文献   

9.
Controlled environment studies were conducted to determine the effects of inoculum density, temperature, leaf wetness and light regime on the infection of linseed by Alternaria linicola. The % cotyledons and leaves with symptoms, and the disease severity (% leaf area with symptoms) increased linearly when the inoculum density increased from 1×103 to 1×105 conidiaml–1. The first symptoms appeared on cotyledons and leaves 4 and 6 days after inoculation, respectively. Eight hours of leaf wetness were sufficient to initiate the disease at 25°C but not at 15°C, when 10-h periods of leaf wetness were required. % leaf area with symptoms was lower at 15°C than that at 25°C irrespective of the leaf wetness periods tested. Interruption of a continuous leaf wetness period by a 12-h dry period, occurring at any time between 1 and 18h after inoculation, decreased the % cotyledons with symptoms and the disease severity, with the greatest reductions (60% and 100%, respectively) being observed when the dry period began 6h after inoculation. A. linicola conidia were able to exploit successive 12-h periods of leaf wetness cumulatively to infect linseed plants. Disease incidence and severity were positively correlated with the dark period following inoculation, but they were negatively related to the length of the initial light period. Our findings suggest that infection of linseed by A. linicola and further development of symptoms can occur under unfavourable environmental conditions.  相似文献   

10.
Blackleg disease (phoma stem canker) of Brassica napus (canola, oilseed rape) is caused by the fungus Leptosphaeria maculans . In some regions of Australia, resistance in oilseed rape cultivars derived from B. rapa subs . sylvestris (e.g. cv. Surpass 400) became ineffective within three years of commercial release. The genetic control of avirulence in L. maculans towards cv. Surpass 400 is described. When Australian field isolates were screened on this cultivar, three phenotypic classes were observed; virulent, intermediate and avirulent. Analysis of crosses between fungal isolates varying in their ability to infect cv. Surpass 400 demonstrated the presence of two unlinked avirulence genes, AvrLm1 and AvrLmS . Complementation of isolates (genotype avrLm1 ) with a functional copy of AvrLm1 , and genotyping of field isolates using a molecular marker for AvrLm1 showed that virulence towards Rlm1 is necessary, but not sufficient, for expression of a virulent phenotype on cv. Surpass 400. Taken together, these data strongly suggest that cv. Surpass 400, with ' sylvestris -derived' resistance, contains at least two resistance genes, one of which is Rlm1 .  相似文献   

11.
Leptosphaeria maculans , the causal agent of phoma stem canker on oilseed rape, is an important pathogen in oilseed rape growing regions of the world, including Australia. Survival of L. maculans and associated mycobiota on oilseed rape stubble buried for 13 months in field soil and in sandy soil was studied under South Australian environmental conditions. Stubble weight decreased significantly by the end of the burial period, more so in field (53·7%) than in sandy soil (22%). Pseudothecia did not develop on stubble buried in field soil and few formed when buried in sandy soil. Moist incubation of stubble following retrieval from both media generated pseudothecia; however, pseudothecial development ceased on stubble that had been buried for 10 and 12 months in field and sandy soil, respectively. In total, 20 and 36 genera of fungi were isolated from stubble before and after burial, respectively. Alternaria spp., L. maculans and Stemphylium botryosum were isolated from 81·7, 70 and 60% of stubble pieces before burial, respectively. Isolation frequency of these species decreased significantly throughout the burial period in both media. Conversely, isolation frequency of Stachybotrys chartarum , Fusarium spp. and Trichoderma spp., having pre-burial frequencies of 26·7, 16·7 and 2·5%, respectively, increased over the burial period regardless of soil type. These findings suggest that inoculum production of L. maculans decreases with the increasing burial duration in field soil over 10 months, before ceasing, and this may be due to associated mycobiota.  相似文献   

12.
In June/July 2001, 2002, 2003 and 2006, regional variation in distribution of the pathogens Leptosphaeria maculans and L. biglobosa that are causally associated with phoma stem canker was surveyed on winter oilseed rape crops in England. In 2001–2003, when isolates from basal cankers were visually identified as L. maculans or L. biglobosa based on cultural morphological characteristics, 70% were L. maculans and 30% L. biglobosa . In 2001, 2002, 2003 and 2006, when amounts of DNA of each species in basal cankers were determined by quantitative PCR, the abundance of L. maculans DNA was greater than that of L. biglobosa DNA in 77% of samples. When regional differences in amounts of L. maculans and L. biglobosa DNA were mapped geostatistically, quantities of L. maculans DNA were greater in cankers from southern England and those of L. biglobosa DNA were greater in northern England. A comparison with geostatistically mapped predictions made using a weather-based model describing stages in development of phoma stem canker epidemics suggested that these differences in Leptosphaeria populations may have been a consequence of differences in temperature after onset of leaf spotting between northern and southern England. Both PCR and morphological evidence suggested that the abundance of L. maculans in England has increased since the last surveys in the 1980s. Implications of these surveys for control of phoma stem canker are discussed.  相似文献   

13.
Effects of temperature on maturation of pseudothecia of Leptosphaeria maculans and L. biglobosa , closely related species which coexist on UK oilseed rape, were investigated. Stages in pseudothecial maturation on naturally infected oilseed rape debris were examined, both in controlled environments (5, 10, 15 or 20°C) under continuous wetness and in natural conditions (debris exposed in September and December 2000, and July, September and November 2002). Pseudothecia sampled weekly were assigned to maturation classes A (asci undifferentiated), B (asci differentiated), C (ascospores differentiated) or D (ascospores mature). Progress in pseudothecial maturation (assessed by time until 50% of pseudothecia reached each class) was similar for L. maculans and L. biglobosa at 15–20°C, but L. biglobosa matured more slowly at < 10°C. Maturation time decreased almost linearly with temperature from 5 to 20°C under continuous wetness but was longer in natural conditions, especially when periods of dry weather occurred. Differences in pseudothecial maturation are likely to contribute to epidemiological differences between L. maculans and L. biglobosa , which may explain their coexistence. It is appropriate to use the degree-day approximation to assess pseudothecial maturation at temperatures between 5 and 20°C, providing debris is wet.  相似文献   

14.
Experiments in controlled environments were carried out to determine the effects of temperature and leaf wetness duration on infection of oilseed rape leaves by conidia of the light leaf spot pathogen, Pyrenopeziza brassicae . Visible spore pustules developed on leaves of cv. Bristol inoculated with P. brassicae conidia at temperatures from 4 to 20°C, but not at 24°C; spore pustules developed when the leaf wetness duration after inoculation was longer than or equal to approximately 6 h at 12–20°C, 10 h at 8°C, 16 h at 6°C or 24 h at 4°C. On leaves of cvs. Capricorn or Cobra, light leaf spot symptoms developed at 8 and 16°C when the leaf wetness duration after inoculation was greater than 3 or 24 h, respectively. The latent period (the time period from inoculation to first spore pustules) of P. brassicae on cv. Bristol was, on average, approximately 10 days at 16°C when leaf wetness duration was 24 h, and increased to approximately 12 days as temperature increased to 20°C and to 26 days as temperature decreased to 4°C. At 8°C, an increase in leaf wetness duration from 10 to 72 h decreased the latent period from approximately 25 to 16 days; at 6°C, an increase in leaf wetness duration from 16 to 72 h decreased the latent period from approximately 23 to 17 days. The numbers of conidia produced were greatest at 12–16°C, and decreased as temperature decreased to 8°C or increased to 20°C. At temperatures from 8 to 20°C, an increase in leaf wetness duration from 6 to 24 h increased the production of conidia. There were linear relationships between the number of conidia produced on a leaf and the proportion of the leaf area covered by 'lesions' (both log10-transformed) at different temperatures.  相似文献   

15.
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).  相似文献   

16.
The survival of Leptosphaeria maculans , which causes phoma stem canker (blackleg), on oilseed rape residues ( Brassica napus ) in South Australia was investigated. Using a quantitative polymerase chain reaction (PCR) assay for L. maculans DNA, the pathogen was mainly detected in the upper 5 cm of the soil profile, including residues on the soil surface. As the size of organic matter particles in the soil decreased, so did the quantity of L. maculans detected in them. To obtain representative data for a field, at least 30 subsamples needed to be collected over the 0·81 ha area studied. In a survey of 49 commercial fields in South Australia, most L. maculans was detected in fields 1 year after oilseed rape had been grown, with less detected after 2 years and negligible amounts 3 years or more after cropping. The diagnostic DNA-based assay for L. maculans reduced the time and cost of studying L. maculans survival in soil and increased the sensitivity and accuracy of results compared with estimates of propagule number of colony-forming units on a semiselective medium.  相似文献   

17.
The effects of temperature on the development of light leaf spot (Pyrenopeziza brassicae) on winter oilseed rape were investigated in controlled-environment experiments. The proportion of conidia which germinated on leaves, the growth rate of germ tubes, the severity of light leaf spot and the production of conidia increased with increasing temperature from 5 to 15 C. The time to 50% germination of conidia and the incubation and latent periods of light leaf spot lesions decreased when temperature increased from 5 to 15°C. At 20°C, however, light leaf spot severity and production of conidia were less and the incubation and latent periods were longer than at 15 C. There were differences between P brassicae isolates and oilseed rape cultivars in the severity of light leaf spot, the production of conidia and the length of the incubation period but not in the length of the latent period. The responses to temperature for lesion severity and incubation and latent periods appeared to be approximately linear over the temperature range 5-15°C and could be quantified using linear regression analysis.  相似文献   

18.
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.  相似文献   

19.
A compartmental model was developed to describe the progress with time of light leaf spot ( Pyrenopeziza brassicae ) on leaves of winter oilseed rape ( Brassica napus ) during the autumn in the UK. Differential equations described the transition between the four compartments: healthy susceptible leaves, infected symptomless leaves, sporulating symptomless leaves and leaves with necrotic light leaf spot lesions, respectively. The model was fitted to data on the progress of light leaf spot on winter oilseed rape at a single site during the autumn of the 1990–1991 season. Model parameters were used to describe rates of leaf appearance, leaf death, infection by airborne ascospores (primary inoculum) and infection by splash-dispersed conidiospores (secondary inoculum). Infection was dependent on sufficient leaf wetness duration. The model also included delay terms for the latent period between infection and sporulation and the incubation period between infection and the appearance of necrotic light leaf spot lesions. This modified SEIR model formulation gave a reasonable fit to the experimental data. Sensitivity analysis showed that varying the parameter accounting for the rate of infection by ascospores affected the magnitude of the curves after the start of the epidemic, whilst including a parameter for conidiospore infection improved the fit to the data. Use of ascospore counts from different sites and different years showed variation in spore release patterns sufficient to affect model predictions.  相似文献   

20.
Ascospores of both A-group and B-group Leptosphaeria maculans germinated at temperatures from 5 to 20°C on leaves of oilseed rape. Germination of ascospores of both groups started 2 h after inoculation and percentage germination reached its maximum about 14 h after inoculation at all temperatures. Both the percentage of A-/B-group ascospores that had germinated after 24 h incubation and germ tube length increased with increasing temperature from 5 to 20°C. Germ tubes from B-group ascospores were longer than those from A-group ascospores at all temperatures, with the greatest difference at 20°C. Hyphae from ascospores of both groups penetrated the leaves predominantly through stomata, at temperatures from 5 to 20°C. A-group ascospores produced highly branched hyphae that grew tortuously, whereas B-group ascospores produced long, straight hyphae. The percentage of germinated ascospores that penetrated stomata increased with increasing temperature from 5 to 20°C and was greater for A-group than for B-group L. maculans after 40 h incubation.  相似文献   

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