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1.
Corynebacterium sepedonicum was detected in symptomless potato stems and tubers with immunofluorescence using monoclonal antibodies specific for the bacterial ring rot pathogen. The concentration of bacterial cells in potato tissue preparations ranged from >500 cells/microscope field to 1 cell per preparation. Symptomless tubers containing ring rot bacteria planted in field plots yielded plants with ring rot symptoms, plants with latent ring rot infections, or plants with no detectable levels of ring rot bacteria. Tubers with the greatest number of bacteria were most likely to develop plants expressing ring rot symptoms, but even some seed tubers with a low number of bacteria developed into plants with symptoms. Some seed tubers with high levels of ring rot bacteria produced plants with only low numbers ofC. sepedonicum.  相似文献   

2.
Summary An indirect sandwich enzyme-linked immunosorbent assay (ELISA) was used to monitor potato stems of three cultivars for the presence of the ring rot pathogen,Clavibacter michiganensis subsp.sepedonicus, during the growing season and in progeny tubers after harvest. The highest ELISA values were obtained with the highest concentration of bacteria used to inoculate seed pieces in all cultivars tested. Low ELISA values were obtained for stems and progeny tubers selected from plants grown from seed inoculated at lower bacterial concentrations. Estimates of bacterial densities in stems and progeny tubers by immunofluorescence indicated that low ELISA values were most probably caused by low bacterial numbers. It is suggested that the sensitivity of ELISA for detecting the ring rot pathogen in potato stems and progeny tubers is a function of the concentration of bacteria in individual seed pieces.  相似文献   

3.
The effectiveness of a tuber incubation method for detection ofErwinia carotovora var.atroseptica andE. carotovora var.carotovora in potato tubers was compared with a lenticel sampling procedure. In the first method, tubers were injured by puncturing lenticels with sterile toothpicks, then wrapped in moist paper towels and polyvinylidene film, and placed in closed chambers flushed with N2. In later experiments, wrapping tubers in two layers of polyvinylidene film and incubation in air was found to be as effective as the single layer of polyvinylidene and incubation in chambers flushed with N2. Isolations were made on a selective crystal violet pectate (CVP) medium from homogenized samples of tissue removed from soft rot lesions developing around injured lenticels. In the second method, 10 lenticels/tuber were aseptically removed with a scalpel and homogenized in distilled water; the suspension was plated on CVP. The first method was less tedious and slightly more effective than the lenticel sampling method. In a preliminary survey, these methods were used to detectErwinia infestations in small samples of certified seed potato tubers from Maine, Minnesota, Montana, New York, North Dakota, and Wisconsin. PectolyticErwinia spp. were detected in at least one sample from each state except Montana. The percentage of tubers withErwinia infestations varied from 0–100% among samples. Characterization ofErwinia isolates showed that bothE. carotovora var.carotovora andE. carotovora var.atroseptica were present. PectolyticErwinia spp. on symptomless potato seed tubers may serve as inoculum sources for blackleg and soft rot diseases.  相似文献   

4.
The latex agglutination test was evaluated for detection of the potato bacterial ring rot pathogen,Corynebacterium sepedonicum. The bacterium was detected in infected potato stems and tubers tested and in inoculated eggplant stems. Test reliability was confirmed using blind tests. A minimum of 106 bacterial cells/ml was requisite for observable agglutination. Nonspecific reactions were not observed from healthy extracts of plant tissues. Nonspecific reactions with other bacteria maintained in pure culture were eliminated by addition of bovine serum albumin to sensitized latex preparations.  相似文献   

5.
Summary Plant-to-plant transmission of the bacterial ring rot (BRR) pathogen,Clavibacter michiganensis subsp.sepedonicus (Spieck. et Kott.) Skapt. et Burkh. of potato (Solanum tuberosum L.), was studied in field trials over a three year period. Healthy and infected seed tubers were planted 35 cm apart. In one treatment, a subsurface barrier was placed between the healthy and infected seed tubers separating the root systems of neighbouring plants. In this treatment, none of 216 plants grown from healthy seed tubers was infected at harvest, as determined by indirect immunofluorescence antibody staining (IFAS) with monoclonal antibodies. In the other treatment, no subsurface barrier was used. In this treatment, two of 368 plants (0.5%) grown from healthy seed tubers were infected at harvest. It is concluded that plant-to-plant transmission may occur but at very low frequency, and is unlikely to play a significant role compared with the potential of transmission by shared potato handling equipment.  相似文献   

6.
Summary Potato stems and tubers grown in the field from seed tubers inoculated withClavibacter michiganensis subsp.sepedonicus, which causes bacterial ring rot, were tested by indirect, sandwich enzyme-linked immunosorbent assay (ELISA) in five laboratories. Correlation between values for each experimental treatment from the five laboratories was greater (r=0.86) than correlation between values for individual samples (r=0.71). When three or more laboratories obtained ELISA values of ≥0.200 for a sample, that sample was presumed to be positive. Conversely, when three or more laboratories obtained ELISA values <0.200, the consensus determination was regarded as negative. The percentage of stem and tuber samples that were in agreement with the consensus ELISA determination varied from 65.5 to 96.7%. Indirect immunofluorescence tests, conducted on the same samples in two laboratories, were consistent with 83.4–91.9% of the consensus ELISA determinations. Presence or absence ofC.m. sepedonicus was confirmed in some samples by an eggplant bioassay and direct isolatiion of the bacterium. The ELISA procedure was well suited for screening large numbers of samples and this study confirms it to be a promising procedure in routine indexing of seed potatoes forC.m. sepedonicus.  相似文献   

7.
The bacterial soft rot potential in potato tubers was affected by differences in temperature between tubers and suspensions ofErwinia carotovora at the time of inoculation by immersion. In general, disease severity after incubation was higher when tuber temperatures were higher as opposed to equal to or lower than that of water containing cells of the bacterium. Disease severity was also greater when tubers were stored at 20° to 23°C in contrast to 4° or 26° to 30°C for 16–18 h prior to immersion. Tubers with a pulp temperature of 8°C were more prone to bruise injury as compared with those at 18°C when struck with a pendulum type bruising instrument. When the injured tubers were inoculated and incubated, most bruises became infected with soft rot bacteria.  相似文献   

8.
The potential of flusulfamide (2’, 4-dichloro- α, α, α, trifluoro-4’-nitro-m-toluenesulfonanilide) as a control agent for bacterial ring rot of potato was evaluated by testing the bactericidal activity of this compound against the causal agent,Clavibacter michiganensis subsp.sepedonicus inin vitro tests, followed by greenhouse and field trials involving treatment of inoculated seed tubers. In thein vitro tests, significant reduction in the size and number ofC. m. sepedonicus colonies was observed with complete inhibition of growth occurring at flusulfamide concentrations of 100 mg/1 or greater. In the greenhouse and field trials, tubers of potato cultivar Russet Burbank were inoculated with the causal organism of bacterial ring rot,C. m. sepedonicus, in one of three different ways (cutting, dipping, injection) and subsequently treated by dipping in an aqueous solution containing varying concentrations of flusulfamide. The “dip” and “cut” methods of inoculation were intended to simulate transmission that would occur during normal production practices, while the “inject” treatment was intended to assess the effect of flusulfamide on existing infections. In both the greenhouse and field experiments, foliar disease incidence and infection rates were reduced, but not eliminated, in plants that were inoculated by cutting or dipping and treated with flusulfamide, whereas plants that had been inoculated by injection showed little effect of treatment with flusulfamide. Based on the results of these experiments, flusulfamide appears to have protective rather than curative properties againstClavibacter michiganensis subsp.sepedonicus.  相似文献   

9.
Apical stem cuttings removed from potato plants (cv. Russet Burbank) infected withCorynebacterium sepedonicum and expressing mild symptoms of bacterial ring rot were demonstrated to be free from the pathogen. This material remained free of ring rot through two additional stem cutting generations and when grown in the field for four successive tuber generations.  相似文献   

10.
A polymerase chain reaction (PCR)-based test for potato late blight (Phytophthora infestans) and pink rot (P.erythroseptica, P. nicotianae) diseases has been developed for use with potato tuber tissue. Primers based on sequence analysis of the ITS2 region of ribosomal DNA of late blight and pink rot pathogens were utilized in PCR assays of inoculated tubers and tubers harvested from plots known to have late blight and/or pink rot. Assays of artificially inoculated Kennebec and Russet Burbank tubers revealed thatP. infestans was detected by PCR as early as 72 h after inoculation and in the absence of visible symptoms. Much higher detection frequencies were obtained by PCR compared with plating on selective medium or placement of tissue in moist chambers. Tubers from plots known to have late blight and/or pink rot were tested using the PCR assay. Assay of late blight lesions showed ca. 80% recovery for late blight-infected tubers from the field. Results indicate that the PCR assay provides a rapid and accurate test for diagnosis of late blight and pink rot in potato tubers.  相似文献   

11.
The ring rot bacterium,Corynebacterium sepedonicum (Spieck. and Kotth.) Skapt. and Burkh., and latent potato viruses (potato virus S and potato virus X) were investigated for their effect on ring rot symptom development on potato plants in the greenhouse and on symptom development and yield of potatoes in the field. Both virus-free (VF) and virus-infected (VI) Red Pontiac stem cuttings root-inoculated with ring rot bacteria in the greenhouse developed typical (T) ring rot symptoms, and symptom severity did not differ between VF and VI plants. In a field study, both VF and VI Russet Burbank seed pieces knife-inoculated with ring rot bacteria produced plants with atypical (A) and T ring rot symptoms as well as a combination of both types. The data suggest that more A than T symptoms develop on VI plants and more T than A symptoms develop on VF plants. Combined infection with the ring rot pathogen and the latent potato viruses resulted in greater yield losses of total and marketable Russet Burbank tubers than infection with the bacterial or viral pathogens alone.  相似文献   

12.
Summary Plants were grown in the field from seed pieces of potato cultivars injected with ring rot bacteria (Corynebacterium sepedonicum). Symptoms were produced with both 18 and 180 colony forming units (cfu) on cvs Norchip and Red Pontiac and with 180 cfu on cv. Belrus plants. Tuber symptoms were detected in all cultivars except Belrus and Teton. Tuber progeny produced plants that developed top symptoms on all cultivars except those of cv. Nooksack, Russet Burbank, and Teton. Tuber progeny of the second crop produced both plants and tubers with symptoms developing only in the cv. Nooksack. In another 3-year experiment, variability in the disease response of selected cultivars and lines was examined following knife-inoculation of tuber seed with a high level of ring rot bacteria. Significant correlations between top and tuber symptoms were detected, but they were not high enough to make unnecessary the examination of both top and tuber symptoms in ring rot disease selection studies. Contribution no. 3879111  相似文献   

13.
Erwinia carotovora var.atroseptica (van Hall) Dye andE. carotovora var.carotovora (Jones) Dye were detected in agricultural soils in Wisconsin using baiting and enrichment techniques. These soft rot bacteria could not be detected in soils of potato fields prior to planting of crops in the spring period using standard soil dilution plating techniques on a crystal violet pectate medium, however. A procedure involving incubation of samples in pectate enrichment broth followed by preparation of smears on slides and treatment with a fluorescent antibody stain specific forE. carotovora var.atroseptica was the most sensitive of the methods tested for detecting the blackleg pathogen. Erwinia carotovora was isolated more frequently during the spring from fields in which potatoes had been grown the previous year than from fields in which other crops had been grown. It was also isolated from potato tubers and stems that had overwintered in the field. The presence ofE. carotovora could not be detected in root zone samples of weed plants using the dilution plating method.  相似文献   

14.
A corky patch syndrome that effects potato tuber tissue may be a later manifestation of pink eye disease of potato although an actual cause and effect relationship between the two diseases has not been firmly established. The causal agent for pink eye of potato is unknown but the disease has been associated with the fungiVerticillium andRhizoctonia and the bacteriumPseudomonas fluorescens. Pink eye and corky patch symptoms on the periderm of affected tubers are accompanied by a band of tissue beneath that is intensely auto-fluorescent when viewed under ultraviolet illumination. Studies to determine the cause of the fluorescent reaction were performed using HPLC and a fluorescence monitor. Three phenylpropanoid compounds, chlorogenic acid, esculin and scopoletin, were found to be involved. It appears that these compounds are produced by tuber tissues in response to pathogen invasion and also as part of the wound healing process. Tissues with corky patch syndrome, silver scurf and root-knot nematode contained high concentrations of chlorogenic acid and esculin and low concentrations of scopoletin. Tissues infected with dry rot were visibly less fluorescent, and contained less chlorogenic acid and esculin, and no detectable level of scopoletin.  相似文献   

15.
The survival of the ring rot bacterium,Corynebacterium sepedonicum, on contaminated surfaces and in infected stems was investigated by root inoculating potato stem cuttings with aqueous suspensions prepared from these materials. The pathogen survived for 24 mo on contaminated surfaces of burlap, kraft paper, and polyethylene plastic held at 12% relative humidity (RH) at either 5 or 20°C. It persisted for less than 14 mo on surfaces held at 94% RH at either temperature. Infectious ring rot bacteria were also recovered from dried, infected potato stems held for 26 mo in an unheated machine shed. These results emphasize the importance of strict sanitation and disinfestation procedures in maintaining potato seed stocks free of bacterial ring rot.  相似文献   

16.
应用PCR技术快速检测马铃薯环腐病菌   总被引：1，自引：0，他引：1  
根据马铃薯环腐病菌16SrDNA基因片段核苷酸序列设计引物(引物1∶5-′TGTACTCGGCCAT-GACGTTGG-3′和引物2∶5-′TACTGGGTCATGTTGGT-3)′,进行马铃薯环腐病菌PCR特异性扩增试验。合成的引物能从马铃薯环腐病菌总基因组DNA和细菌纯培养,以及人工接种和自然发病的马铃薯块茎中特异性扩增环腐病菌16SrDNA基因片段1046bp。该试验结果为马铃薯环腐病的鉴定、检测及病害流行学研究提供了新的技术和方法。  相似文献   

17.
The bacterial ring rot disease of potato (Corynebacterium sepedonicum) can be detected in seed potato lots by serological tests with greater accuracy than by visual field inspections. Indexing tests can make use of highly specific immunofiuorescence probes, enzyme-linked immunosorbent assays, and perhaps other procedures. Sensitivity and specificity of these procedures need to be determined in order to estimate the confidence with which ring rot can be detected in a seed lot. In addition to the laboratory tests, the sampling strategy determines the efficacy and sensitivity of the indexing procedure. Samples might consist of stems, leaf petioles, or tubers in single or composite units. Disease incidence and number of plants sampled also determine the confidence level with which ring rot can be detected in a potato seed lot. Although research has been conducted on many aspects of ring rot testing procedures, further work needs to be done. Nevertheless, it is anticipated that indexing potato seed lots for the presence of ring rot will be a significant factor in control and eradication of the disease.  相似文献   

18.
Leak is a post-harvest disease of potato tubers manifested by a wet, spongy rot that can quickly lead to complete tuber decay. In North America, leak is most commonly caused byPythium ultimum var.ultimum Trow, although a number of otherPythium spp. have been found to be pathogenic to potato tubers. We report that an isolate identified by DNA sequencing asP. sylvaticum Campbell & Hendrix was pathogenic to potato tubers. The organism produced a dark, hollow cavity in tubers that was much drier in texture than typical symptoms of leak. Symptomology was similar in tubers of the cultivars Yukon Gold, Green Mountain, and Russet Burbank. Studies to complete Koch’s postulates confirmed thatP. sylvaticum could be reisolated from diseased tissue and produced the same disease symptoms when isolates were subsequently re-inoculated into healthy tubers. This is the first report ofP. sylvaticum causing rot in potato tubers.  相似文献   

19.
Soft rot resistance in potato (Solanum tuberosum L.) tubers can be determined by inoculating tuber slices with soft rotErwinia species. Questions have been raised in the literature concerning the reliability of the tuber slice method. The objectives of this study were to 1) examine the statistical assumptions underlying the analysis of variance for different response variables as measures of soft rot resistance using the tuber slice method of evaluation; 2) estimate the sample sizes necessary to detect specified differences with power 0.83; and, 3) choose the “best” variable for measuring resistance to soft rot based on valid statistical analysis and minimal sample size. Slices from fifteen tubers from each of three cultivars (Atlantic, Norchip, Superior) were inoculated withErwinia carotovora subsp.atroseptica, E. carotovora subsp.carotovora orE. chrysanthemi and incubated at two different temperatures (20 C and 30 C) for 48 hrs. The test was conducted on two dates. Tuber samples were sliced and weighed prior to inoculation and after the macerated tissue was removed following a 48 hr incubation period. The maximum diameter of macerated tissue, actual weight loss, four measures of proportional weight loss, and various transformations of these variables were analyzed. The “best” response variables for measuring resistance to soft rot by the tuber slice method were the diameter of the macerated tissue and the square root transformation of one of the measures of proportional weight loss. No differences were found among the cultivars nor theErwinia subsp. for either of these response variables, and the cultivar ×Erwinia subsp., cultivar × temperature and cultivar ×Erwinia subsp. × temperature interactions were not significant. However, there were significant differences between the incubation temperatures, and theErwinia subsp. × temperature interaction was significant for both response variables. Estimates of sample sizes necessary to find a 20% difference in main effect and interaction effect means were calculated.  相似文献   

20.
Bacterial ring rot-infected tuber and stem tissue reacted withCorynebacterium sepedonicum antiserum in agar immunodiffusion (AID). The AID test was as effective as immunofluorescence (IF) in confirming ring rot in potato tubers submitted for diagnosis. Precipitin bands developed in AID with tuber preparations that contained, in a 1:10 dilution of the preparation, at least 100 IF positive cells/microscope field with a 100 X oil objective. Only a low percentage of preparations with less than 100 IF positive cells were positive with AID. Samples inoculated withC. sepedonicum strains which produced non-mucoid colony types on YGM medium did not consistently develop precipitin bands in AID which suggests that the usefulness of the test may be limited where non-mucoid strains occur naturally.  相似文献   

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