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1.
Detailed studies were conducted on the distribution of Pectobacterium carotovorum subsp. carotovorum and Dickeya spp. in two potato seed lots of different cultivars harvested from blackleg-diseased crops. Composite samples of six different tuber sections (peel, stolon end, and peeled potato tissue 0.5, 1.0, 2.0 and 4.0 cm from the stolon end) were analysed by enrichment PCR, and CVP plating followed by colony PCR on the resulting cavity-forming bacteria. Seed lots were contaminated with Dickeya spp. and P. carotovorum subsp. carotovorum (Pcc), but not with P. atrosepticum. Dickeya spp. and Pcc were found at high concentrations in the stolon ends, whereas relatively low densities were found in the peel and in deeper located potato tissue. Rep-PCR, 16S rDNA sequence analysis and biochemical assays, grouped all the Dickeya spp. isolates from the two potato seed lots as biovar 3. The implications of the results for the control of Pectobacterium and Dickeya spp., and sampling strategies in relation to seed testing, are discussed.  相似文献   

2.
Suspected Dickeya sp. strains were obtained from potato plants and tubers collected from commercial plots. The disease was observed on crops of various cultivars grown from seed tubers imported from the Netherlands during the spring seasons of 2004–2006, with disease incidence of 2–30% (10% in average). In addition to typical wilting symptoms on the foliage, in cases of severe infection, progeny tubers were rotten in the soil. Six strains were characterised by biochemical, serological and PCR-amplification. All tests verified the strains as Dickeya sp. The rep-PCR and the biochemical assays showed that the strains isolated from blackleg diseased plants in Israel were very similar, if not identical to strains isolated from Dutch seed potatoes, suggesting that the infection in Israel originated from the Dutch seed. The strains were distantly related to D. dianthicola strains, typically found in potatoes in Western Europe, and were similar to biovar 3 D. dadanti or D. zeae. This is the first time that the presence of biovar 3 strains in potato in the Netherlands is described. One of the strains was used for pathogenicity assays on potato cvs Nicola and Mondial. Symptoms appeared 2 to 3 days after stem inoculation, and 7 to 10 days after soil inoculation. The control plants treated with water, or plants inoculated with Pectobacterium carotovorum, did not develop any symptoms with either method of inoculation. The identity of Dickeya sp. and P. carotovorum re-isolated from inoculated plants was confirmed by PCR and ELISA.  相似文献   

3.
Dickeya species (formerly Erwinia chrysanthemi) cause diseases on numerous crop and ornamental plants world‐wide. Dickeya spp. (probably D. dianthicola) were first reported on potato in the Netherlands in the 1970s and have since been detected in many other European countries. However, since 2004–5 a new pathogen, with the proposed name ‘D. solani’, has been spreading across Europe via trade in seed tubers and is causing increasing economic losses. Although disease symptoms are often indistinguishable from those of the more established blackleg pathogen Pectobacterium spp., Dickeya spp. can initiate disease from lower inoculum levels, have a greater ability to spread through the plant’s vascular tissue, are considerably more aggressive, and have higher optimal temperatures for disease development (the latter potentially leading to increased disease problems as Europe’s climate warms). However, they also appear to be less hardy than Pectobacterium spp. in soil and other environments outside the plant. Scotland is currently the only country in Europe to enforce zero tolerance for Dickeya spp. in its potato crop in an attempt to keep its seed tuber industry free from disease. However, there are a number of other ways to control the disease, including seed tuber certification, on‐farm methods and the use of diagnostics. For diagnostics, new genomics‐based approaches are now being employed to develop D. dianthicola‐ and ‘D. solani’‐specific PCR‐based tests for rapid detection and identification. It is hoped that these diagnostics, together with other aspects of ongoing research, will provide invaluable tools and information for controlling this serious threat to potato production.  相似文献   

4.
Potato blackleg is a seedborne disease that can cause significant economic losses for growers. Disease development depends mainly on two drivers, namely seed inoculum and local climatic conditions. To better establish the relationship between these two drivers, blackleg development was monitored in Swiss field trials at multiple locations from 2010 to 2013 involving three sets of naturally infected seed lots planted in each of three locations. The seed lot itself was thereby the most important factor explaining differences in disease development, rather than environmental factors. In a further on-farm project conducted at various locations in Switzerland and southern Germany from 2013 to 2015, the implementation of a seed-testing procedure was investigated. A total of 177 seed lots were tested for natural latent infection with soft rot Pectobacteriaceae and the corresponding blackleg incidence was tracked in 242 fields. The reliability of the relationship between latent infection and field incidence was found to be strongly linked to the bacterial species. Dickeya spp. field infection could be predicted with an acceptable reliability, whereas Pectobacterium carotovorum subsp. brasiliense, even when detected as latent tuber infection, was not consistently expressed as visual blackleg. Moreover, commonly found mixed latent infections with several bacterial species made it even harder to predict which bacteria would cause blackleg symptoms. Finally, variability in the reliability of seed testing may also be explained by differences in local farming practices. These trials over several years with naturally infected potato seed highlight the usefulness and limits of seed testing to manage blackleg.  相似文献   

5.
Biochemical characterisation of Dickeya strains isolated from potato plants and river water samples in Finland showed that the majority of the strains were biovar 3. They thus resembled the strains recently isolated from potato in the Netherlands, Poland and Israel and form a new clade within the Dickeya genus. About half of the Finnish isolates resembling strains within this new clade were virulent and caused wilting, necrotic lesions and rotting of leaves and stems. Similar symptoms were caused by D. dianthicola strains isolated from one potato sample and from several river water samples. Frequently, the rotting caused by the Dickeya strains was visible in the upper parts of the stem, while the stem base was necrotic from the pith but hard and green on the outside, resulting in symptoms quite different from the blackleg caused by Pectobacterium atrosepticum. The presence of Dickeya in the symptomatic plants in the field assay was verified with a conventional PCR and with a real-time PCR test developed for the purpose. The virulent Dickeya strains reduced the yield of individual plants by up to 50% and caused rotting of the daughter tubers in the field and in storage. Management of Dickeya spp. in the potato production chain requires awareness of the symptoms and extensive knowledge about the epidemiology of the disease.  相似文献   

6.
Potato blackleg, caused by Pectobacterium and Dickeya species, is one of the most significant bacterial diseases affecting potato production globally. Although it is generally accepted to be a seedborne disease, the processes underlying the spread of disease largely remain unknown. Spatial point pattern analysis was applied to blackleg occurrence in seed potato crops in Scotland during the period of 2010–2013 (approximately 8000 blackleg‐affected crops), to assess whether its distribution was random, regular or aggregated, and the spatial scales at which these patterns occurred. Blackleg‐affected crops derived from mother stocks with symptoms were omitted from the analyses in order to examine the statistical evidence for horizontal transmission of blackleg. The pair correlation function was used to test for global spatial autocorrelation, and results indicated significant (< 0·05) clustering of incidence at a wide range of spatial scales. Strength of clustering (degree of aggregation) among blackleg‐affected crops was notably larger at spatial scales of 25 km or less. A hot‐ and coldspot analysis was performed to test for local spatial autocorrelation, and statistically significant clusters of high and low values of disease were found across the country. These analyses provide the first quantitative evidence of localized and large‐scale spatial clustering of potato blackleg. Understanding the mode(s) of inoculum dispersal will be important for developing new management strategies that minimize host–pathogen contacts in potato and numerous other crops affected by pathogenic Pectobacterium and Dickeya species.  相似文献   

7.
Plant pathogenic enterobacteria in the genera Pectobacterium and Dickeya (formerly classified as Erwinia) were isolated from diseased potato stems and tubers. The isolated bacteria were identified as P. atrosepticum, P. carotovorum and pathogens in the genus Dickeya with PCR tests. Furthermore, Dickeya strains were isolated from river water samples throughout the country. Phylogenetic analysis with 16S-23S rDNA intergenic spacer sequences suggested that the Dickeya strains could be divided into three groups, two of which were isolated from potato samples. Phylogenetic analysis with 16S rDNA sequences and growth at 39°C suggested that one of the groups corresponds to D. dianthicola, a quarantine pathogen in greenhouse cultivation of ornamentals, while two of the groups did not clearly resemble any of the previously characterised Dickeya species. Field trials with the strains indicated that D. dianthicola-like strains isolated from river samples caused the highest incidence of rotting and necrosis of potato stems, but some of the Dickeya strains isolated from potato samples also caused symptoms. The results showed that although P. atrosepticum is still the major cause of blackleg in Finland, virulent Dickeya strains were commonly present in potato stocks and rivers. This is the first report suggesting that Dickeya, originally known as a pathogen in tropical and warm climates, may cause diseases in potato in northern Europe.  相似文献   

8.
A protocol for the specific detection and quantification of ‘Candidatus Liberibacter solanacearum’ in carrot seeds using real‐time PCR was developed. The bacterium was detected in 23 out of 54 carrot seed lots from 2010 to 2014, including seeds collected from diseased mother plants. The average total number of ‘Ca. L. solanacearum’ cells in individual seeds ranged from 4·8 ± 3·3 to 210 ± 6·7 cells per seed from three seed lots, but using propidium monoazide to target live cells, 95% of the cells in one seed lot were found to be dead. Liberibacter‐like cells were observed in the phloem sieve tubes of the seed coat and in the phloem of carrot leaf midrib from seedlings. The bacterium was detected as early as 30 days post‐germination, but more consistently after 90 days, in seedlings grown from PCR positive seed lots in an insect‐proof P2 level containment greenhouse. Between 12% and 42% of the seedlings from positive seed lots tested positive for ‘Ca. L. solanacearum’. After 150 days, symptoms of proliferation were observed in 12% of seedlings of cv. Maestro. ‘Candidatus Liberibacter solanacearum’ haplotype E was identified in the seeds and seedlings of cv. Maestro. No phytoplasmas were detected in seedlings with symptoms using a real‐time assay for universal detection of phytoplasmas. The results show that to prevent the entry and establishment of the bacterium in new areas and its potential spread to other crops, control of ‘Ca. L. solanacearum’ in seed lots is required.  相似文献   

9.
Possibilities for biocontrol of biovar 3 Dickeya sp. in potato were investigated, using bacteria from rotting potato tissue isolated by dilution plating on nonselective agar media. In a plate assay, 649 isolates were screened for antibiosis against Dickeya sp. IPO2222 and for the production of siderophores. Forty‐one isolates (6·4%) produced antibiotics and 112 isolates (17·3%) produced siderophores. A selection of 41 antibiotic‐producing isolates and 41 siderophore‐producing isolates were tested in a potato slice assay for control of the Dickeya sp. Isolates able to reduce rotting of potato tuber tissue by at least 50% of the control were selected. Isolates were characterized by 16S rDNA analysis as Bacillus, Pseudomonas, Rhodococcus, Serratia, Obesumbacterium and Lysinibacillus genera. Twenty‐three isolates belonging to different species and genera, 13 producing antibiotics and 10 producing siderophores, were further characterized by testing acyl‐homoserine lactone (AHL) production, quorum quenching, motility, biosurfactant production, growth at low (4·0) and high (10·0) pH, growth at 10°C under aerobic and anaerobic conditions and auxin production. In replicated greenhouse experiments, four selected antagonists based on the in vitro tests were tested in planta using wounded or intact minitubers of cv. Kondor subsequently inoculated by vacuum infiltration with an antagonist and a GFP (green fluorescent protein)‐tagged biovar 3 Dickeya sp. strain. A potato endophyte A30, characterized as S. plymuthica, protected potato plants by reducing blackleg development by 100% and colonization of stems by Dickeya sp. by 97%. The potential use of S. plymuthica A30 for the biocontrol of Dickeya sp. is discussed.  相似文献   

10.
Dickeya spp. and Pectobacterium atrosepticum are major pathogens of potato. Current methods to detect these soft-rotting bacteria require separate identification steps. Here we describe a simple method allowing simultaneous detection of both pathogens based on multiplex PCR. The sensitivity of the primer sets was first examined on purified genomic DNA of the type strains Dickeya chrysanthemi 2048T and P. atrosepticum 1526T. The specificity and detection limits of the primer sets were successfully tested on 61 strains belonging to various Dickeya and Pectobacterium species, on artificially inoculated and on naturally contaminated potato plants. This new method provides a gain in time and materials, the main advantages for large-scale processes such as pathogen-free seed certification.  相似文献   

11.
It is well established that the pectinolytic bacteria Pectobacterium atrosepticum (Pca) and Dickeya spp. are causal organisms of blackleg in potato. In temperate climates, the role of Pectobacterium carotovorum subsp. carotovorum (Pcc) in potato blackleg, however, is unclear. In different western and central European countries plants are frequently found with blackleg from which only Pcc can be isolated, but not Pca or Dickeya spp. Nevertheless, tubers vacuum-infiltrated with Pcc strains have so far never yielded blackleg-diseased plants in field experiments in temperate climates. In this study, it is shown that potato tubers, vacuum-infiltrated with a subgroup of Pcc strains isolated in Europe, and planted in two different soil types, can result in up to 50% blackleg diseased plants.  相似文献   

12.
Long‐term survival of Acidovorax citrulli in citron melon (Citrullus lanatus var. citroides) seeds was investigated. Citron melon seed lots infected with A. citrulli were generated in the field by inoculating either the pistils (stigma) or pericarps (ovary wall) of the female blossoms. Seventeen A. citrulli isolates from 14 different haplotypes belonging to two different groups (group I and II) were used for inoculation. After confirming that 100% of seed lots were infected, they were stored at 4°C and 50% RH for 7 years. After storage, the viability of A. citrulli cells from individual lots was determined by plating macerated seeds on semiselective medium as well as growing seeds for 14 days and scoring for bacterial fruit blotch symptoms. The type of A. citrulli isolate (group I or group II) used did not significantly influence bacterial survival. However, A. citrulli survival was significantly greater in seed lots generated via pistil inoculation (52·9 and 29·4%) than via pericarp inoculation (23·5 and 17·6%). Repetitive extragenic palindrome (rep)‐PCR on A. citrulli isolated from citron melon seed lots after storage displayed similar fingerprinting patterns to those of the reference strains originally used for blossom inoculation, indicating that cross‐contamination did not occur. The results indicate that A. citrulli may survive/overwinter in citron melon seeds for at least 7 years and bacterial survival in seed was influenced more by method of blossom inoculation than by the type of bacterial isolate.  相似文献   

13.
Bacterial soft rots are a serious limitation to the production of orchids and other horticultural plants. Here, the characterization of causative bacteria isolated from Phalaenopsis orchids showing symptoms, from a commercial production site, is reported. The most commonly isolated bacteria were identified as Dickeya spp. Partial sequencing of 16S rDNA, fliC and dnaX showed diversity among the isolates and divided the isolates into two groups, with greatest similarity to previously reported undefined Dickeya lineages from orchids (UDL‐3 and UDL‐4). Two isolates (B16, S1) were sequenced using next‐generation sequencing, which has provided draft genomes of these two isolates for further studies (Ali? et al., 2015 ). Newly developed fliC‐based lineage‐specific quantitative real‐time PCR assays were used to distinguish among the lineages and to assess their relative abundances in diseased tissues. Virulence and aggressiveness comparison tests in vivo on Phalaenopsis orchids, potato plants and witloof chicory leaves indicated high virulence and extreme maceration potential of these novel Dickeya isolates, compared to a reference panel of other Dickeya spp. Pantoea cypripedii (formerly Pectobacterium cypripedii), which has previously been reported as a soft rot pathogen of orchids, was not detected, and isolates obtained from culture collections did not cause symptoms on artificially infected Phalaenopsis orchids.  相似文献   

14.
Tsror  Leah  Aharon  M.  Erlich  Orly 《Phytoparasitica》1999,27(3):215-226
Potato seed tubers are imported to Israel from northern Europe and planted in spring; tubers harvested early from the spring crop are used as seed for the autumn crop. Although only seed lots registered as certified are imported, a previous survey (1984–1994) indicated that most imported lots were affected by latent or active infections caused byErwinia carotovora,Streptomyces scabies, Rhizoctonia solani, Fusarium spp. andSpongospora subterranae. The survey was extended until 1998, and included additional pathogens:Ralstonia solanacearum,Helminthosporium solani, Colletotrichum coccodes andVerticillium dahliae. Most of these pathogens were also monitored in domestic seed tubers, and are reported for the first time. Brown rot was not observed in any of the imported lots. Blackleg and soft rot caused byErwinia spp. were detected in most of the imported lots; however, less than 7% of the lots were contaminated at high levels, while approximately 65% were contaminated at moderate levels. Common scab was detected in most of the imported lots; 51% of the imported lots were contaminated at moderate or high levels, whereas only 6.5% of the domestic seed lots were contaminated at these levels. Black scurf was detected in most of the imported lots; on average, 47.3%, 44.2% and 1.4% of the lots were contaminated at low, moderate and high levels, respectively, and only 7.1% were disease-free. In contrast, most of the domestic lots were either disease-free (45.4%) or had a low disease incidence (37.3%). Only 16.7% of the lots were moderately infected and 0.2% were highly contaminated. Silver scurf was observed in most of the imported lots during all years of the survey, with no differences among the producing countries; on average, 22.7%, 66.1% and 7.5% of the lots were contaminated at low, moderate and high levels, respectively, and only 3.7% were disease-free. Most of the domestic lots (76%) were disease-free and only 6.6% were infected at moderate or high levels. Black dot was observed in a considerable portion of the shipments from Holland during all years of the survey, particularly in 1998, when 34% of the lots were infected. The shipments from France and Germany were infected at low levels, except in 1998, when 19% and 11% of the lots, respectively, arrived infected. In shipments from Scotland and Ireland low incidences of the disease were observed in 1994 and 1995. In the domestic lots, black dot incidence was low (<2.4%) except in 1996, when 11% of the lots were infected.V. dahliae was monitored only in domestic seed tubers. The incidence of disease-free lots was 56–64%, whereas in 20–30% of the lots the level of infection was <5%, and in 6–16% of the lots the level was >5%. The survey findings demonstrate transmission of seedborne pathogens; most of these pathogens can become established in the soil and eventually cause severe outbreaks of disease in potatoes grown in Israel. http://www.phytoparasitica.org posting May 16, 1999.  相似文献   

15.
A total of 109 samples of potato seed tubers imported to Jordan from France, Netherlands and Denmark during the 2007/2008 growing seasons were surveyed for 10 different diseases caused by fungi and fungus‐like organisms to determine the prevalence of different tuberborne diseases and to evaluate the efficacy of the current visual examination procedures in detecting different potato diseases. The results demonstrated that most potato seed tuber lots imported to Jordan were infected with one or more of the following potato pests: Colletotrichum coccodes (black dot) (66.0%), Rhizoctonia solani (black scurf) (42.4%), Helminthosporium solani (silver scurf) (92.6%), and Spongospora subterranea (powdery scab) (13.8%). Just over a quarter (25.7%) of all lots examined in this study were found to exceed the acceptance limits for one or more potato diseases according to the Jordanian standards. This study suggests that the current visual examination procedure is not adequate to detect all tuberborne diseases of potato and to estimate their incidence and severity precisely.  相似文献   

16.
Dickeya and Pectobacterium are responsible for causing blackleg of plants and soft rot of tubers in storage and in the field, giving rise to losses in seed potato production. In an attempt to improve potato health, biocontrol activity of known and putative antagonists was screened using in vitro and in planta assays, followed by analysis of their persistence at various storage temperatures. Most antagonists had low survival on potato tuber surfaces at 4 °C. The population dynamics of the best low-temperature tolerant strain and also the most efficient antagonist, Serratia plymuthica A30, along with Dickeya solani as target pathogen, was studied with TaqMan real-time PCR throughout the storage period. Tubers of three potato cultivars were treated in the autumn with the antagonist and then inoculated with D. solani. Although the cell densities of both strains decreased during the storage period in inoculated tubers, the pathogen population was always lower in the presence of the antagonist. The treated tubers were planted in the field the following growing season to evaluate the efficiency of the bacterial antagonist for controlling disease incidence. The potato endophyte S. plymuthica A30 protected potato plants by reducing blackleg development on average by 58.5% and transmission to tuber progeny as latent infection by 47–75%. These results suggest that treatment of potato tubers with biocontrol agents after harvest can reduce the severity of soft rot disease during storage and affect the transmission of soft rot bacteria from mother tubers to progeny tubers during field cultivation.  相似文献   

17.
Dickeya strains isolated in Israel in 2006–2010 were characterized by dnaX sequence analysis, pulsed‐field gel electrophoresis (PFGE), biochemical assays and pectolytic activity, and found to be homogeneous: most of them could be classified as ‘Dickeya solani’. Of the 34 strains isolated from imported seed tubers or potato plants grown from imported seed, 32 were typed as ‘D. solani’ and only two were characterized as Dickeya dianthicola. Biovar typing indicated that all ‘D. solani’ strains were biovar 3. ‘Dickeya solani’ strains were most closely related to Dickeya dadantii subsp. dieffenbachiae according to PFGE and dnaX analyses and both species exhibited high pectolytic activity. Expression levels of two putative virulence genes, pelL (encoding a pectic enzyme) and dspE (encoding a type III effector) were significantly induced in ‘D. solani’ strains isolated from potato plants or tubers grown in hot climates such as the Negev region in Israel, compared to those isolated from seed tubers imported from the Netherlands, France or Germany. Results of this study support the hypothesis that ‘D. solani’ strains isolated in Israel are also clonal; however, they appear to be more virulent than strains isolated in Europe.  相似文献   

18.
J. MARTIN 《EPPO Bulletin》1998,28(1-2):177-182
Diagnosis of tobacco rattle tobravirus (TRV) in potato is difficult because of the existence of non-encapsidated NM-type strains, because similar symptoms are caused by potato mop-top furovirus and physiological internal rust spot and because serological tests are inefficient. The aim of this study was first to compare ELISA and PCR tests. The former was found to be ineffective, so only the PCR technique was in a survey of numerous seed-potato lots (200-tuber samples) from different EU countries. A large disparity was found according to geographical origin, with infection levels of German, Dutch and French seed potatoes of 10.3, 8.5 and 0.6%, respectively. There is a real risk that planting tubers with such infection levels will newly contaminate soil. Besides, TRV is not considered in the EU texts on marketing of seed potatoes, so there is no legal recourse if seed-potato lots are found to be TRV-infected.  相似文献   

19.
Common scab is one of the most important soil‐borne diseases of potato and is difficult to control. Selection of potato breeding lines for resistance to common scab is also cumbersome due to environmental factors influencing symptom development and an erratic spatial distribution of the scab pathogens (Streptomyces spp.) in the field. The bacterial phytotoxin thaxtomin A, which causes scab symptoms, can be used to screen large numbers of potato seedlings for tolerance in vitro, but few studies have investigated whether the results correspond to resistance to common scab observed in the field. In this study, 120 F1 potato progeny from a single cross were screened in vitro by exposing the seedlings to thaxtomin A added to the culture medium. Eighteen genotypes were selected based on high sensitivity or tolerance using shoot growth as the criterion, multiplied in vitro, and tested for resistance to common scab caused by S. turgidiscabies and S. scabies in a glasshouse and in three different fields. Evaluation of ca. 6500 tubers showed that the 18 potato genotypes differed in scab indices and disease severity (P < 0·0001). The relative shoot height in vitro (thaxtomin A used at 0·5 μg mL?1) and the scab index in the field showed significant correlation (rs = ?0·463, P = 0·0528, n = 18), also consistent with the results obtained under controlled conditions in the glasshouse. Hence, the in vitro bioassay may be used to discard scab‐susceptible genotypes and elevate the overall levels of common scab resistance in the potato breeding populations.  相似文献   

20.
Stem rot symptoms caused by pectinolytic bacteria of Genus Pectobacterium and Genus Dickeya, which are commonly referred to as blackleg, strongly impact the quality of seed potato production in most European countries. Several biotic and abiotic factors, such as cultivar susceptibility, isolate aggressiveness, mother tuber infection density and a wide range of soil-related and climatic factors have been identified in the literature as having an effect on blackleg development. The aim of this study was to identify which biotic and/or abiotic factors are most critical to the development of blackleg in the field. In Switzerland, the predominant species have belonged to Genus Dickeya as far back as 1992, which is why this study only investigates blackleg symptoms induced by Dickeya isolates. Seven field trials, in which inoculated tubers were planted, were conducted during a 3-year period and the number of blackleg-diseased plants was counted. Multiple regression analysis was used in order to determine the factors that had the greatest impact on two different variables: (i) periods between emergence of the plant and disease outbreak and (ii) overall blackleg incidence throughout the growing season. The results of this analysis have revealed that environmental factors, such as evapotranspiration and soil moisture, explain about half of the variability in the number of days before disease outbreak, and the total number of diseased plants is widely dependent upon cultivar susceptibility and isolate aggressiveness.  相似文献   

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