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1.
The characteristics of 36 isolates of Leptosphaeria maculans originating from cv. Picra were investigated using cultural, pathogenicity and isozyme analyses in comparison with six known isolates of A- and B-groups. Picra-related isolates were similar to the three known B-group isolates from Brassica napus using a cultural test. Two known B-group isolates and 29 of the 36 Picra-related isolates tested were aggressive on cv. Picra, whereas all the three known A-group isolates were not. All the 42 isolates tested were aggressive on the B. napus – B. juncea recombinant line MX and on B. napus cv. Westar. Phosphoglucose isomerase analysis showed two different patterns, separating A-group known (fast band) from B-group known isolates (slow band). All cv. Picra isolates displayed the slow band. Considerable differences in isoesterase patterns were found between known A- and B-group isolates. Isolates recovered from cv. Picra displayed six different electrophoretic patterns, including that of the known B-group isolates. Clusters of the 42 isolates, generated by UPGMA analysis based on similarity coefficients of electrophoretic types, matched the previous classification into A- and B-groups, although two atypical Picra-related isolates with a singular pattern clustered with the A-group. The importance of these isolates for blackleg epidemiology and resistance breeding is discussed.  相似文献   

2.
ABSTRACT The Brassica napus-B. juncea recombinant line (MX), resistant to Leptosphaeria maculans, was produced by interspecific crosses and bears one gene (Jlm1) from the B. juncea B genome. We investigated whether this new resistance was race specific by characterizing protection against a large sample of L. maculans isolates. The pathogenicity of 119 isolates of L. maculans comprising 105 A-group isolates and 14 B-group isolates was studied at the cotyledon stage under controlled conditions using the MX line, the susceptible B. napus cultivar Westar, and the resistant B. juncea cultivar Picra. All but one of the isolates were pathogenic on 'Westar'. Only 3 of the 105 A-group isolates caused very mild symptoms on 'Picra'. Two of these strains were isolated from the MX line and the other from Sinapis arvensis. The other 102 strains caused hypersensitive-type responses. Most B-group isolates were pathogenic on 'Picra'. There were differences in pathogenicity among A-group isolates tested on the MX line, whereas all B-group isolates were pathogenic on this line. A-group isolates obtained from the MX line were more frequently pathogenic on the MX line than those obtained from B. napus cultivars. One isolate from S. arvensis infected the MX line. These results suggest that the resistance of the MX line is unlikely to be durable. Thus, the new resistance gene Jlm1 should probably be used in association with other sources of resistance, in plant breeding schemes, to prevent the breakdown of this resistance.  相似文献   

3.
The influence of competitive effects between two isolates, of the number of infection sites on cotyledons and of host polygenic resistance on the systemic growth of Leptosphaeria maculans , the cause of phoma stem canker in oilseed rape ( Brassica napus ), were investigated. Controlled-condition experiments were conducted with two oilseed rape doubled haploid lines, one susceptible and the other with a high level of polygenic resistance, inoculated via wounded cotyledons with conidial suspensions obtained from two isolates. Expression of cankers in plants was enhanced by exposing inoculated plants to low temperature (6°C) followed by warm temperature (20°C). The fungus was detected by PCR amplifications of three minisatellite markers in all stems with visible canker symptoms and also in the stems of 14 of the 59 plants without visible cankers on the hypocotyls. Disease severity increased with the number of infection sites on cotyledons: in one of the three replicate experiments, the mean external necrosis length on the hypocotyl ranged from 6·47 to 35·3 mm for one and eight infections sites on cotyledons, respectively. The probability of an isolate reaching the hypocotyl from inoculated cotyledons decreased with increasing competing inoculum load on cotyledons: for instance, for isolate A290v it decreased from 1 when inoculated alone to 0·28 when coinoculated with six drops of competing isolate P27d. Polygenic resistance significantly reduced disease incidence and severity. For instance, in one of the three replicate experiments, disease incidence ranged from more than 74% in susceptible plants to 16% in resistant ones, while mean external necrosis length was up to 35·3 and 6·5 mm on susceptible and on resistant plants, respectively. This study offers new possibilities for assessing levels of polygenic resistance to stem canker in B. napus and studying the aggressiveness of L. maculans isolates.  相似文献   

4.
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   

5.
Pairwise combinations were carried out between the eight isolates originating from each of three asci of the B-group of Leptosphaeria maculans . Some of the pairings produced mature pseudothecia when incubated on wheat straws at 18°C and under a 12-h photoperiod of blue light. Isolates within a given tetrad could be attributed to one of two sexual compatibility groups, on the basis of combinations of isolates leading to fertile crosses. Within each tetrad, mating type segregated in a 1:1 ratio, suggesting the existence of one mating type gene ( MAT1 ) with two alleles MAT 1–1 and MAT 1–2. Under these same conditions, each individual isolate from an A-group ascus originating from Brassica juncea produced pseudothecia with one or other of two compatible A-group testers originating from B. napus . However, attempts to mate these eight A-group single-ascospore isolates with two B-group single-ascospore isolates of opposite mating types remained unsuccessful. This work is the first successful report of in vitro sexual reproduction between B-group isolates, and provides evidence for bipolar heterothallism in B-group isolates of L. maculans . The implications of this mating protocol for genetic studies of B-group isolates and the consequences of this work for the taxonomy of L. maculans are discussed.  相似文献   

6.
Verticillium wilt of oilseed rape ( Brassica napus ) is caused primarily by Verticillium longisporum and has become a serious problem in northern Europe. In order to evaluate whether V. longisporum and V. dahliae differ in their interaction with oilseed rape, phenotypical and molecular assessments were made. Oilseed rape plants for fungal assessments were inoculated with V. longisporum and V. dahliae via root-dipping and samples were taken from roots, stems, leaves, flowers, pods and seeds during plant development. The infection by V. longisporum was found to start mainly in lateral roots and root-hairs, followed by colonization of the xylem vessels and extensive spread in stems and leaves, whereas V. dahliae infected the main roots and remained in the region below the cotyledon node of the plants. Re-isolation studies, together with PCR analysis of samples taken from early growth stages through to fully ripe plants, showed that the onset of flowering was a critical phase for V . longisporum to colonize plants. No seeds infected with V. longisporum were found. Mycelial growth from V. dahliae but not V. longisporum was significantly reduced on media containing tissue from a low glucosinolate B. napus genotype compared with growth on media containing tissue from a high glucosinolate cultivar. The results of this study suggest that V. longisporum favours B. napus as host and that the transition from the vegetative to the generative phase is of importance for the spread of the fungus in oilseed rape plants.  相似文献   

7.
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   

8.
The response of cotyledons of oilseed rape ( Brassica napus ssp. oleifera ) accessions to infection by isolates of Peronospora parasitica under controlled conditions was assessed on a 0–7 scale (disease reaction). In interactions scored 0–3, 4–5 and 6–7, the host was considered resistant, partially resistant and susceptible, respectively. Accession RES-26, selected from the spring oilseed rape cultivar Janetzkis, was partially resistant to isolate R1 and resistant to isolate P003 of P. parasitica , which distinguishes it from three previously described differential response groups ('A', 'B' and 'C') of accessions in B. napus . The resistance of RES-26 to isolate P003 seemed to be conditioned by a single, partially dominant gene and the resistance of RES-02, which belongs to group 'A' (resistant to R1 and P003), by two independent partially dominant genes. The gene for resistance to P003 in RES-26 is either closely linked, allelic or identical to one of the two genes for resistance in RES-02. Resistance of RES-02 to R1 is conditioned by a single, incompletely dominant gene. The genes for resistance to isolates R1 and P003 in RES-02 are either closely linked, allelic or identical. The cotyledonary leaves of each seedling responded independently when inoculated simultaneously each with a different isolate of the pathogen.  相似文献   

9.
European Journal of Plant Pathology - Phoma stem canker / blackleg is an internationally important disease of Brassicas including B. napus (oilseed rape, OSR), caused by multiple genetic subclades...  相似文献   

10.
ABSTRACT Verticillium wilt caused by the vascular fungal pathogen Verticillium longisporum is one of the most important pathogens of oilseed rape (Brassica napus sp. oleifera) in northern Europe. Because production of this major oilseed crop is expanding rapidly and no approved fungicides are available for V. longisporum, long-term control of the disease can only be achieved with cultivars carrying effective quantitative resistance. However, very little resistance to V. longisporum is available within the gene pool of oilseed rape, meaning that interspecific gene transfer from related species is the only possibility for broadening levels of resistance in current varieties. The amphidiploid species B. napus can be resynthesized by crossing the two progenitor species Brassica oleracea and Brassica rapa, hence resistant accessions of these two diploid species can be used as resistance donors. In this study a total of 43 potential B. rapa and B. oleracea resistance donors were tested with regard to their reaction to a mixture of two aggressive V. longisporum isolates, and resistances from diverse lines were combined by embryo rescue-assisted interspecific hybridization in resynthesized rapeseed lines. Progenies from crosses of the two B. rapa gene bank accessions 13444 and 56515 to the B. oleracea gene bank accessions BRA1008, CGN14044, 8207, BRA1398, and 7518 showed a broad spectrum of resistance in pathogenicity tests. Of 45 tested resynthesized lines, 41 lines exhibited a significantly higher level of resistance than the moderately V. longisporum-tolerant oilseed rape cultivar Express. These lines represent a promising basis for the combination of different resistance resources in new varieties.  相似文献   

11.
Seedlings of 13 cultivars of oilseed rape ( Brassica napus ) were inoculated with 10 isolates of Leptosphaeria maculans. Three virulent isolates were identified which gave a resistant reaction on some cultivars normally considered susceptible. Further experiments to compare these with standard isolates confirmed the existence of a differential host-pathogen interaction. The breeding line 78/271, plus the cultivars Quinta, Garant, Norli and possibly Elvira differentiated the isolates tested while Jet Neuf, Rafal, Primor, Doral, Hercules, Rapora, Liraglu and Erra did not. The relevance of these findings to breeding for canker resistance in oilseed rape is discussed.  相似文献   

12.
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   

13.
Pretreatment of the first true leaves of oilseed rape plants ( Brassica napus cv. Bristol) with menadione sodium bisulphite (MSB) locally and systemically induced resistance, as shown by reduced lesion size and number, to infection by the fungal pathogen Leptosphaeria maculans , the causal agent of stem canker. Using a known systemic activator of salicylic acid-dependent PR-1 induction, acibenzolar- S -methyl (BTH; S -methylbenzo[1,2,3]thiadiazole-7-carbothiate) as a comparison, real-time PCR expression analysis of genes encoding a pathogenesis-related protein 1 ( PR-1 ) and an ascorbate peroxidase ( APX ) demonstrated a systemic enhancement of APX expression in MSB-pretreated plants, with no effect on PR-1 expression, suggesting augmented reactive oxygen species production in MSB-pretreated plants. The results demonstrate MSB to be an effective resistance activator in oilseed rape, and potentially useful for the control of stem canker.  相似文献   

14.
Sclerotinia stem rot (SSR) of oilseed rape (OSR, Brassica napus), caused by Sclerotinia sclerotiorum, is a serious problem in the UK and worldwide. As fungicide‐based control approaches are not always reliable, identifying host resistance is a desirable and sustainable approach to disease management. This research initially examined the aggressiveness of 18 Sclerotinia isolates (17 S. sclerotiorum, one S. subarctica) on cultivated representatives of B. rapa, B. oleracea and B. napus using a young plant test. Significant differences were observed between isolates and susceptibility of the brassica crop types, with B. rapa being the most susceptible. Sclerotinia sclerotiorum isolates from crop hosts were more aggressive than those from wild buttercup (Ranunculus acris). Sclerotinia sclerotiorum isolates P7 (pea) and DG4 (buttercup), identified as ‘aggressive’ and ‘weakly aggressive’, respectively, were used to screen 96 B. napus lines for SSR resistance in a young plant test. A subset of 20 lines was further evaluated using the same test and also in a stem inoculation test on flowering plants. A high level of SSR resistance was observed for five lines and, although there was some variability between tests, one winter OSR (line 3, Czech Republic) and one rape kale (line 83, UK) demonstrated consistent resistance. Additionally, one swede (line 69, Norway) showed an outstanding level of resistance in the stem test. Resistant lines also had fewer sclerotia forming in stems. New pre‐breeding material for the production of SSR resistant OSR cultivars relevant to conditions in the UK and Europe has therefore been identified.  相似文献   

15.
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   

16.
ABSTRACT Leptosphaeria maculans, the causal agent of stem canker of oilseed rape (Brassica napus), develops gene-for-gene interactions with oilseed rape, and four L. maculans avirulence (AVR) genes (AvrLm1, AvrLm2, AvrLm4, and alm1) were previously genetically characterized. Based on the analysis of progeny of numerous in vitro crosses between L. maculans isolates showing either already characterized or new differential interactions, this work aims to provide an overview of the AVR genes that may specify incompatibility toward B. napus and the related species B. juncea and B. rapa. Two novel differential interactions were thus identified between L. maculans and B. napus genotypes, one of them corresponding to a complete resistance to European races of L. maculans. In both cases, a single gene control of avirulence was established (genes AvrLm3 and AvrLm7). Similarly, a single gene control of avirulence toward a B. rapa genotype, also resistant to European L. maculans isolates, was demonstrated (gene AvrLm8). Finally, a digenic control of avirulence toward B. juncea was established (genes AvrLm5 and AvrLm6). Linkage analyses demonstrated that at least four unlinked L. maculans genomic regions, including at least one AVR gene cluster (AvrLm1-AvrLm2-AvrLm6), are involved in host specificity. The AvrLm3-AvrLm4-AvrLm7 region may correspond either to a second AVR gene cluster or to a multiallelic AVR gene.  相似文献   

17.
Verticillium longisporum is one of the major pathogens of oilseed rape (Brassica napus; genome AACC, 2n = 38) in Europe. Current European cultivars possess only a low level of resistance against V. longisporum, meaning that heavy infection can cause major yield losses. The aim of this study was to identify quantitative trait loci (QTL) for resistance against V. longisporum as a starting point for marker-assisted breeding of resistant cultivars. Resistance QTL were localized in a segregating oilseed rape population of 163 doubled haploid (DH) lines derived by microspore culture from the F1 of a cross between two B. napus breeding lines, one of which exhibited V. longisporum resistance derived by pedigree selection from a resynthesized B. napus genotype. A genetic map was constructed comprising 165 restriction fragment length polymorphism, 94 amplified fragment length polymorphism and 45 simple sequence repeats (SSR) markers covering a total of 1,739 cM on 19 linkage groups. Seedlings of the DH lines and parents were inoculated with V. longisporum isolates in four greenhouse experiments performed in Sweden during autumn 1999. In three of the experiments the DH lines were inoculated with a mixture of five isolates, while in the fourth experiment only one of the isolates was used. The intention was to simulate four different environments with variable disease pressure, while still maintaining uniform conditions in each environment to enable reliable disease scoring. The disease index (DI) was calculated by scoring symptoms on a total of 21 inoculated plants per line in comparison to 21 noninoculated plants per line. Using the composite interval mapping procedure a total of four different chromosome regions could be identified that showed significant QTL for resistance in more than one environment. Two major QTL regions were identified on the C-genome linkage groups N14 and N15, respectively; each of these QTL consistently exhibited significant effects on resistance in multiple environments. The presence of flanking markers for the respective QTL was associated with a significant reduction in DI in the inoculated DH lines.  相似文献   

18.
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   

19.
Williams  & Fitt 《Plant pathology》1999,48(2):161-175
Stem canker or blackleg of brassicas, caused by Leptosphaeria maculans , is one of the most damaging diseases of winter oilseed rape in the UK. Airborne ascospores, released in autumn and winter, initiate leaf infections which may lead to colonization of the petiole and, later in the season, formation of stem lesions and cankers. Although isolates of the pathogen differ in ability to cause damaging stem cankers, this is not readily apparent from leaf spotting or stem lesion symptoms. However, several cultural, biochemical and genetic characteristics appear to be associated with the ability to form damaging stem cankers and isolates can be assigned to one of two groups, termed A and B, on the basis of differences in these characteristics. To investigate the relationship between leaf spotting symptoms and subsequent stem canker formation, and to improve understanding of the epidemiology of this pathogen, it is desirable to differentiate between the stem canker forming A group and the less damaging B group of L. maculans . Characterization of isolate type is also important in seed testing and crop breeding programs, particularly in countries such as Canada and Poland where the A type is not ubiquitous. This article reviews methods, including plant assays, assessments of growth characteristics in vitro , isozyme analyses, secondary metabolite profiling, serology, and nucleic acid analyses, that can be used to differentiate the A and B groups.  相似文献   

20.
The growth of two isolates of the downy mildew fungus Peronospora parasitica , one obtained from cauliflower ( Brassica oleracea ) and the other from oilseed rape ( B. napus ) was assessed in their respective hosts of origin, and also in the alternative combination. Both isolates were capable of infecting either host, but there were marked contrasts in the time course and extent of mycelial development, the amounts of associated host-cell necrosis, and eventual intensity of sporulation. Oilseed rape, which was partially resistant to the isolate from cauliflower, exhibited extensive necrosis of mesophyll cells, in conjunction with reduced mycelial development, and delayed and reduced sporulation by the pathogen. The isolate from oilseed rape was virulent on both host species. Pathogenesis in the susceptible combinations was accompanied by large increases in electrolyte leakage, and increased activity of the enzymes β-glucosidase, ribonuclease, and peroxidase. Effects on chlorophyll content were variable and activities of acid phosphatase and acid phosphodiesterase were unaffected. Electrophoretic analyses of extracts from fungal sporangia and infected seedlings indicated that the large increases in β-glucosidase were of pathogen origin, while evidence from inhibitor studies suggested that enhanced ribonuclease activity was due to a new post-infectional form of the enzyme. The significance of these findings is discussed in relation to pathogenesis and host resistance mechanisms.  相似文献   

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