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1.
The susceptibility of 24 potato cultivars to soft rot caused by Erwinia carotovora subsp. atroseptica was assessed in a simplified form of a tuber slice test described previously. Freshly made wounds on tuber slices were inoculated with drops (0–02 ml) containing 108, 107 or 106 cells/ml and incubated aerobically for 3 days at 15° or 20°C. At 20°C differences between cultivars were clear at all concentrations but at 15°C less so at the lowest concentration unless incubation time was extended. The rank order of cultivar susceptibility was similar to that found in previous seasons.  相似文献   

2.
The thermal death points of Erwinia carotovora subsp. atroseptica and subsp. carotovora were determined in relation to duration of heat treatment, age of culture and culture medium. No isolates cultured in liquid media survived heating at 53°C for 5 min while those on solid media were killed by heating at 54°C for 10 min. After immersing naturally contaminated potato tubers for 10 min in water at 55°C, Erwinia could not be detected. The same treatment of naturally or artificially contaminated seed tubers gave complete absence of blackleg infection in the field and decreased the amounts of powdery scah(Spongospora subterranea) and black scurf (Rhizoctonia solani) on progeny tubers.  相似文献   

3.
The resistance of eight potato cultivars to tuber soft rot caused by E. carotovora subsp. atroseptica was assessed in 2 years using three different test methods. Similar cultivar resistance rankings were obtained for any one method within a year and between years for two methods (single site, infectivity titration), but not for the third (vacuum infiltration). However, the ranking of cultivars differed for the three methods. Ranking was not affected by inoculating the cortex or the more susceptible medullary tissue, or by assessing rotting in terms of infection frequency or lesion size, but it was affected by oxygen concentration during incubation. Differences among cultivars were greater when inoculated tubers were incubated anaerobically than when incubated with 5% oxygen. There was no relationship between the relative susceptibilities of cultivars to tuber soft rot in storage in January/February and those of mother tubers after planting.  相似文献   

4.
In South Africa during the 1988 season, a wilt disease appeared in seed-potato crops. The disease was initially characterized by a one-sided wilt of the upper leaves, and in its advanced stage was indistinguishable from verticillium, fusarium and pseudomonas wilts. A basal stem rot was occasionally associated with wilted plants when wet, humid conditions prevailed. Erwinia carotovora ssp. carotovora and E. chrysanthemi were isolated from wilted plants, but E. carotovora ssp. carotovora was the predominant pathogen. Stem inoculations in the glasshouse showed that both pathogens were capable of causing wilt symptoms. An indication was found that E. chrysanthemi was more virulent at a higher temperature range (28-32°C) and E. carotovora ssp. carotovora at a lower temperature range (20-25°C). It was found that the potting compost, irrigation water and some parent tubers used for the propagation of plantlets at the Foundation Seed Scheme, Eastern Transvaal, were contaminated with E. carotovora ssp. carotovora.  相似文献   

5.
彩色马蹄莲欧文氏菌PCR检测   总被引：1，自引：0，他引：1  
采用聚合酶链式反应(PCR)技术检测彩色马蹄莲欧文氏菌纯培养和彩色马蹄莲样品,并用分离培养技术加以验证。结果表明,PCR能特异地检测出所有10个彩色马蹄莲欧文氏菌菌株,并证实了昆明地区侵染彩色马蹄莲的细菌为胡萝卜软腐欧文氏菌(Erwiniacarotovora subsp.carotovora)。PCR与分离培养技术检测结果基本一致,但总体上PCR检测阳性率稍高于分离培养检测的发病率。接种马铃薯、大白菜24h后接种点处均出现明显软腐症状。该项技术具有更高的灵敏度,适用于彩色马蹄莲种苗的检测和病害流行学研究。  相似文献   

6.
Soft rot erwiniae are a group of notorious plant pathogens for which currently available detection methods are inadequate. Based on the polymerase chain reaction, specific and sensitive detection of Erwinia carotovora subsp. atroseptica and E. chrysanthemi in potato tubers has been achieved. The composition of the PCR primers used in two specific detection systems is based on identification of the consensus of sequences of metalloprotease-coding genes present in soft rot erwiniae. Bacterial DNA was extracted from the potato tuber matrix by differential centrifugation in order to avoid interference of potato-derived compounds with the performance of the PCR assay. The PCR assay jjerformed with the E. carotovora subsp. atroseptica specific primer set was found to be capable of distinguishing E. carotovora subsp. atroseptica from all other Erwinia species and the closely related subspecies E. carotovora subsp. carotovora. With the E. chrysanthemi specific primer set, agarose gel electrophoresis is required for unequivocal differentiation between E. chrysanthemi and other erwiniae. Combined with the efficient extraction procedure, the assay allowed specific detection of less than 103 culturable erwiniae per tuber. The specificity and sensitivity of the assay were not reduced in the presence of a 100-fold excess of DNA from both related and unrelated bacteria. This PCR-based method for detection of erwiniae in potato tubers provides a relatively fast and sensitive alternative to routinely applied serological methods.  相似文献   

7.
The relationship between number of viable cells of Erwinia carotovora subsp. atroseptica on inoculated potato seed tubers and blackleg development was investigated in 2 years for five cultivars grown in the contrasting climates of Scotland and Israel. Blackleg, and to a lesser extent non-emergence, increased with higher numbers of bacteria on the seed tubers at planting. This relationship was also found for several commercial seed stocks of one cultivar naturally contaminated with different numbers of E. carotovora subsp. atroseptica.The threshold number of bacteria necessary for the development of blackleg declined during the growing season and was also higher for the cultivar Pentland Crown in comparison with the others. In general, yield declined linearly with blackleg incidence and there was a 0.8% reduction in yield for every 1 % blackleg at 13 weeks after planting. Yield loss was positively related to the incidence of blackleg late in the season, whereas the relationship between yield loss and the incidence of non-emergence was poor.  相似文献   

8.
Contamination of seed potato tubers by Erwinia carotovora subsp. atroseptica is widespread with the bacteria usually sited superficially in lenticels and suberized wounds. As seed contamination level is related to blackleg incidence, seed health is best assessed by determining the number of cells of E. c. atroseptica per mL of tuber-peel extract. The relative specificity, sensitivity and ease of use of four recently developed microbiological, immunological and molecular methods to detect and/or quantify tuber contamination are discussed in relation to the testing of commercial seed stock. Sensitivities of all four methods are at or below the threshold level for blackleg development (< 103 cells mL-1), but there are differences regarding their specificity and ease of use. Three of them allow enumeration of most live cells of the bacterium, using specific monoclonal and polyclonal antibodies against the predominant serogroup I: (a) immunomagnetic separation of E. c. atroseptica before viable count on a selective-diagnostic growth medium, crystal violet pectate, (b) immunofluorescence staining and counting of colonies in pour-plate medium in tissue culture plates and (c) enrichment of the bacterium in peel-extract dilutions directly in microtitre plates prior to DAS-ELISA. In the fourth method, both live and dead cells are detected, but not quantified, by PCR amplification of target sequences using specific primers for E. c. atroseptica regardless of serogroup.  相似文献   

9.
In Israel field infections of potato plants by Erwinia chrysanthemi are characterized by wilting of the leaves followed by total desiccation of the plants. These symptoms are indistinguishable from those caused by Verticillium dahliae or those that develop during the normal process of plant senescence. Diagnosis of E. chrysanthemi in the spring-sown (February) crop in Israel is difficult because all three conditions often appear at approximately the same time, late in the growing season in May when the air temperature exceeds 25°C. The symptoms of E. chrysanthemi infection were reproduced in the field when potato seed tubers, tested and found to be contaminated at a low level with E. carotovora pv. carotovora , were inoculated with a strain of E. chrysanthemi isolated from a diseased potato plant. When plants in a growth cabinet at 30°C were stem-inoculated with E. chrysanthemi , similar symptoms developed when the relative humidity was low ( c . 80%). Presence of the disease only on plants grown from seed contaminated with E. chrysanthemi and not from uncontaminated seed suggests that the bacterium is seed borne, as is E. carotovora pv. atroseptica , the blackleg pathogen.  相似文献   

10.
Potato tubers of five cultivais were incubated at five temperatures between 2 and 20°C for 1 month and then inoculated with Phoma exigua var. foveata and stored at 5°C. Larger rots usually developed on tubers previously incubated at the higher temperatures but cultivar differences were greatest where tubers had been incubated at 2°C. In another experiment, rot size was related to the duration of incubation at 20°C before inoculation.
In a series of experiments, tubers were stored at different temperatures after inoculation; other factors (e.g. wound type or r.h.) were also investigated in some of these. Combining the results of all experiments a linear decrease of % wounds infected (logit transformation) with storage temperature was demonstrated. The size of rots was not simply related to temperature, but tended to increase over the range 2–10°C and then stabilized or declined at higher temperatures. Relative humidity treatments of 75% v. 95% did not consistently affect either the incidence of rotting or rot size. The other factors investigated did not usually interact significantly with temperature.
The relevance of the results for resistance testing is discussed and a simple equation relating disease incidence to inoculum levels, type of damage and temperature is proposed.  相似文献   

11.
A PCR-RFLP test based on a pectate-lyase encoding gene permits the detection of several Erwinia carotovora subspecies, but requires complete DNA extraction. This paper reports on the suitability of a simplified PCR-RFLP protocol to characterise E. carotovora strains and on the performance of PCR, using the same primers, to detect the atroseptica subspecies in substrates of epidemiological significance. A collection of 140 strains from various hosts and geographical origins was characterised for biochemical traits and PCR-RFLPs. PCR performed on boiled bacterial suspensions yielded an amplification product of 434 bp in 109 of the 140 strains. None of the E. carotovora subsp. betavasculorum strains was amplified, even after complete DNA extraction. RFLPs of the PCR product yielded 24 groups, 3 of which were new. Twenty one groups were specific to one subspecies. Several strains biochemically similar to E. carotovora subsp. atroseptica, but growing at 37 °C, showed PCR-RFLP profiles characteristic of E. carotovora subsp. carotovora. Phenetic and cladistic analyses gave three main domains, not strictly related to hosts or geographical origins. The atroseptica (RFLP groups 1 and 2) and wasabiae (group 21) subspecies constituted one of the domains, despite clustering distantly from one another. Host specialisation and molecular homogeneity suggest a clonal structure within these subspecies. Conversely, E. carotovora subsp. odorifera, despite its limited host range and geographical distribution, and E. carotovora subsp. carotovora showed great molecular diversity, spreading respectively across five and 19 RFLP groups. These two subspecies shared RFLP groups 4, 5 and 6. The tree nodes in the phenograms showed a low robustness when bootstrapping the data matrix. PCR coupled with a 48h enrichment step in a polypectate-rich medium improved detection thresholds of E. carotovora subsp. atroseptica (1.5.102- 1.5.103 bacteria/ml in leaves, stems, and tuber peel extracts to 4.107 bacteria/ml in wash water) relative to either immunomagnetic separation coupled with PCR or DAS-ELISA (2.105 in plant samples to 2.107 bacteria/ml in wash water).  相似文献   

12.
Hélias  rivon  & Jouan 《Plant pathology》2000,49(1):33-42
Transmission of pectinolytic Erwinia species from infected mother tubers to daughter tubers has been studied mainly through detection tests, carried out at harvest, on limited samples of tubers produced by plants grown from artificially inoculated mother tubers. However, detection has not been performed on samples collected at different stages of crop development, in order to follow the contamination progress in different organs through the plants to the progeny tubers. In this study the bacterial contamination of progeny tubers was investigated by detecting Erwinia carotovora ssp. atroseptica in different symptomless plant organs (stolons, stems, progeny tubers) and in the parts with or without symptoms of diseased stems, collected at various stages of crop development. Infection levels in below- and above-ground organs of plants of two cultivars differing in their resistance to Erwinia, infected by either vacuum infiltration or sand wounding, were monitored throughout the growing season and at harvest using DAS-ELISA and PCR. Detection tests showed that healthy organs from symptomless plants were less frequently contaminated than symptomless organs from diseased plants, and that stolons were precociously and more frequently contaminated than stems and daughter tubers, irrespective of the health of the plant. Stem infections were shown to progress latently in the stem, bacteria usually being recovered 10–15 cm past visible lesions. In many cases, typical aerial stem-rot symptoms could be related to this upward movement of bacteria from the infected mother tuber. Daughter tubers without symptoms were shown to be frequently contaminated, usually at heel ends, suggesting internal contamination from mother tuber to progeny.  相似文献   

13.
Hélias  rivon  & Jouan 《Plant pathology》2000,49(1):23-32
Infection of seed tubers by pectinolytic Erwinia species can lead to the development of various symptoms during vegetative growth of potato crops, including non-emergence of plants, chlorosis, wilting, haulm desiccation and typical blackleg. The relationships between types of symptoms and yield are poorly documented, and are investigated by following the development of symptoms in potato plants grown under field conditions from seed tubers artificially inoculated with E. carotovora ssp. atroseptica ( Eca ) , and measuring the yield of each plant. Symptoms were classified into five main types (non-emergence, wilting/chlorosis, blackleg, haulm desiccation and plant death). Each plant was scored for types of symptom on four successive dates; plants without visible symptoms were scored as healthy. The method of inoculation and inoculum concentration proved major factors for the subsequent development of symptoms. Disease development was more severe after vacuum infiltration of bacteria into seed tubers than after shaking tubers in contaminated sand. Disease usually progressed from chlorosis and/or wilting to partial or total desiccation on a given plant. Yield losses varied according to symptom type, but the relationship between symptoms recorded and yield also depended on scoring dates. The data suggest that the beginning of tuber growth might be the most suitable stage for predicting yield losses from symptom observations. In both cultivars studied (Bintje, highly susceptible, and Désirée, moderately resistant), the yield of symptomless plants growing from inoculated seed tubers was significantly less than that of control plants, indicating that the presence of bacteria on the seed tuber was detrimental, even in the absence of visible symptoms. Differences in symptom expression in the field between cultivars matched the level of visible infection of tubers at harvest, as Bintje tubers showed a higher incidence of rot than Désirée tubers.  相似文献   

14.
To investigate the ability of black dot symptoms to develop on infected potato tubers during storage, the growth of Colletotrichum coccodes was followed in vitro on malt agar at temperatures ranging from 5–27°C, and in vivo on artificially infected potato tubers kept at 5, 10 and 15°C. In vitro , 13 isolates from different geographical origins grew at all temperatures tested; growth started with a delay of 10 days at 5°C and of 4 days at 10°C, and was fastest at 27°C. All isolates had similar growth patterns and produced conidia and sclerotia at all temperatures. Minitubers were successfully infected at 5, 10 and 15°C by depositing either a mycelial plug or a drop of conidial suspension on the tuber surface. Sclerotia were observed after 7 days at the point of inoculation. Symptoms extended in all cases, although more slowly at 5 and 10 than at 15°C. Latent infections were detected in up to 21% of tubers without black dot symptoms at harvest. These results show that latent infections by C. coccodes are probably quite frequent, and that the pathogen is able to develop at low temperatures in controlled conditions. This suggests that black dot symptoms can increase during storage if stores are not adequately managed.  相似文献   

15.
Phytophthora infestans is able to produce oospores in leaves of potato and tomato plants after inoculation with a mixture of Al and A2 mating-type isolates. Various conditions for oospore formation were analysed. Under controlled conditions, oospores were produced in potato leaves at temperatures ranging from 5 to 25° C. In leaves of potato cultivar Bintje incubated at 15°C, oogonia and antheridia were observed 6 days after inoculation and thick-walled oospores appeared 3-4 days later. In field experiments oospores were found in leaves and stems of potato cultivars Bintje, Irene and Pimpernel and in leaves, stems and fruits of tomato cultivar Moneymaker within 2 weeks after inoculation. A bioassay was developed to test the survival of oospores in soil under various conditions. To determine whether late-blight infections derived from infectious soil were caused by oospwres, DNA fingerprinting was performed. DNA fingerprint probe RG-57 was suitable for distinguishing asexual progeny from recombinant progeny arising from soil-borne oospores. We demonstrated survival of viable, infectious oospores of P. infestans in soil during the winter of 1992–93. Oospores were not infectious from soil exposed to temperatures of 40°C or higher but in the range 35°C to as low as – 80°C for 48 h, oospores survived.  相似文献   

16.
A time-saving and cost-effective polymerase chain reaction (PCR)-based method was developed for species-specific detection of the scab pathogens ( Streptomyces scabies and S. turgidiscabies ) prevalent in potato ( Solanum tuberosum ) in northern Scandinavia. Species specificity of primers was verified using a collection of previously characterized Streptomyces strains isolated from potato scab lesions in Finland and Sweden. A total of 1245 scab lesions was tested from potato cvs Matilda and Sabina grown in the field in two geographic regions of Finland in 2000 and 2001. Freshly harvested or stored potato tubers were incubated at room temperature (18–21°C) under humid conditions for a few days. Bacterial growth was collected from scab lesions for DNA isolation and PCR. The two scab pathogens were detected in the same potato fields, tubers and scab lesions. The relative incidence of S. scabies was high in freshly harvested tubers but was much lower than that of S. turgidiscabies following storage. Both pathogens were seed-transmitted in Matilda and Sabina after 24 weeks of storage at 4°C.  相似文献   

17.
During the winters of 1977/78 and 1978/79 tubers of cv. Arran Banner naturally contaminated with Phoma exigua var, foveata were kept at either high or low humidity and at 5, 10, 15 or 20 C. During the 1978/79 winter cv. Majestic tubers were stored under similar conditions at 5 and 10°C only. At monthly intervals tubers from each lot were damaged to encourage gangrene development and returned to their storage condition except the Majestic tubers which were switched from high to low humidity or vice versa and returned to the appropriate temperature. No gangrene rots developed in tubers kept at 1 5 or 20°C at either humidity. At 5 and 10°C fewer rots developed at high humidity compared with low humidity including Majestic tubers kept Initially at high humidity. Inoculum levels in soil on tubers were not markedly affected by storage conditions.  相似文献   

18.
An improved method for detection, quantification and classification of soft rot bacteria associated with potato seed tubers, plant material, soil and water has been developed. The method is based on the use of a modified version of the crystal violet pectate selective medium (CVP), enrichment cultures under anaerobic conditions using pectate as the sole carbon source for recovery improvement, and the quantitative estimation ofErwinia spp. by employing a new solid medium - most probable number (MPN) method. The use of this method enabled an improvement in the recovery and identification of specificErwinia spp. in mixed populations. This was done by incubating CVP plates — used for the MPN counting — at three different temperatures (15, 28 and 39°C). These combined techniques were used for estimating low level populations at less than one cell per gram or ml tested ofErwinia carotovora subsp.carotovora, E. carotovora subsp.atroseptica, andE. chrysanthemi.  相似文献   

19.
To examine whether the pathogenic bacterium, Erwinia carotovora subsp. carotovora, causal agent of soft rot of Chinese cabbage (Brassica campestris L., pekinensis group), can overwinter in plant debris and soil and serve as inoculum the following year, we monitored field populations of rifampicin-resistant, phage-sensitive strains of the bacterium. Chinese cabbage (cv. Matsushima Kohai W1116) were planted in field soil in pots that were sunk into the field on Aug. 2, 1996 and eventually reduced to one plant per pot. Outer petioles of the plants were inoculated with mixture of 13 bacterial strains of E. carotovora subsp. carotovora on Sept.5, 1996. After the soft rot spread throughout the plant, the diseased plant was buried in the potted soil. New seeds were sown in the pots on April 30, 1997, and the disease was observed in June and July. The bacterial strains were re-isolated from the potted soil, diseased tissue and rhizosphere soil by the dilution plating method on modified Drigalski's medium containing 100 ppm rifampicin and by the enrichment technique. In addition to rifampicin resistance, phage sensitivities of some of the re-isolated strains were identical to those of the strains buried in the soil with the diseased plant in the previous year. From these results, some of the 13 strains overwintered in the soil and infested plant tissue and acted as primary inoculum the following year. The frequency of re-isolation varied among the strains, perhaps because of competition among the strains, differences in epidemiological behavior and stabilizing selection among the strains, and the presence of different ecotypes of the organism. Received 21 July 2000/ Accepted in revised form 19 September 2000  相似文献   

20.
ABSTRACT Late blight (Phytophthora infestans), pink rot (Phytophthora erythroseptica), leak (Pythium ultimum), dry rot (Fusarium sambucinum), and soft rot (Erwinia carotovora subsp. carotovora and subsp. atroseptica) are particularly damaging diseases of stored potato tubers worldwide. In this study, we present a methodology to detect and quantify the causal agents of the five aforementioned diseases from whole potato tubers, using real-time quantitative-polymerase chain reaction. Six primer pairs were designed to amplify targets smaller than 150-bp DNA in single copy protein-coding gene targets of each of the pathogens and the potato host. Using a large collection of pure culture DNA samples, all primer pairs specifically detected the DNA target in the intended pathogenic species. Amplification efficiencies over a five-log dilution series ranged between 95 and 100% and were unaffected by the presence of large amounts of host DNA. The detection level of the primers reached 0.5 pg of target DNA. Pathogens were detected in 100 pg of total DNA extracted from 170 to 250 g of tubers, 4 days after inoculation, regardless of the presence of symptoms. The presence of P. erythroseptica, Pythium ultimum, or E. carotovora was also detected in 1 ng of DNA extracted from potato tubers collected from a commercial storage facility. This study provides the first step in a methodology to predict the storability of potato tubers following harvest.  相似文献   

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