Semiselective Agar Medium for Isolation of<i>Clavibacter michiganense</i>subsp.<i>michiganense</i>from Tomato Seed

Fatmi, M.; Schaad, N. W.

doi:10.1094/phyto-78-121

Cited by 59 publications

(41 citation statements)

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“…Since it is possible that differences in virulence result from variations in the ability to colonize the host plant, the bacterial titers in tomato plants infected with strain NCPPB382 and its derivatives were determined at various times after infection with a semiselective medium for C. michiganensis which suppresses growth of saprophytic bacteria and fungi (10). As a control we included strain NCPPB3123 in the experiment, a strain which is apathogenic and has lost the ability to effectively colonize tomato plants.…”

Section: Resultsmentioning

confidence: 99%

“…For EPS isolation, bacteria were grown in yeast extract medium (24). For reisolation of bacteria from infected plants, a modified SCM medium (10) 16,000 rpm (25,000 x g) for 30 min. The supernatant was first passed through a cellulose-acetate filter (pore size, 0.45 ,tm) to remove the remaining bacterial cells (steril filtration) and then passed through an Amicon XM50 filter to remove components with molecular masses of 50 kDa or lower.…”

Section: Methodsmentioning

confidence: 99%

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Evidence for plasmid-encoded virulence factors in the phytopathogenic bacterium Clavibacter michiganensis subsp. michiganensis NCPPB382

et al. 1993

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The tomato pathogen Clavibacter michiganensis subsp. michiganensis NCPPB382, which causes bacterial wilt, harbors two plasmids pCM1 (27.5 kb) and pCM2 (72 kb). After curing of the plasmids, bacterial derivatives were still proficient in the ability to colonize the host plant and in the production of exopolysaccharides but exhibited a reduced virulence. When one of the two plasmids is lost, there is a significant delay in the development of wilting symptoms after infection and a plasmid-free derivative is not able to induce disease symptoms. By cloning of restriction fragments of both plasmids in the plasmid-free strain CMM100, two DNA fragments which restored the virulent phenotype were identified. Further analysis suggested that a fragment of plasmid pCM1 encodes an endocellulase which is involved in the expression of the pathogenic phenotype.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Evidence for plasmid-encoded virulence factors in the phytopathogenic bacterium Clavibacter michiganensis subsp. michiganensis NCPPB382

et al. 1993

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“…An isolate of Cmm of known high pathogenicity (obtained from the pathogen collection of the Institute of Plant Protection in Poznań) was used for artificial inoculations. Bacteria were grown on a semiselective agar medium according to Fatmi and Schaad (1988). In the spring trial, the pathogen was introduced into the rockwool culture system 11 days after placing tomato transplants onto rockwool production slabs by injecting 5 ml of water suspension of Cmm inoculum (2 x 10 8 cfu/ml) into rockwool starter cubes at a point just under the drip irrigation emitters.…”

Section: Methodsmentioning

confidence: 99%

Attempts at Biological Control of Clavibacter michiganensis subsp. michiganensis On Rockwool-Grown Greenhouse Tomatoes

Slusarski¹

2008

Journal of Fruit and Ornamental Plant Research

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Attempts at Biological Control ofClavibacter michiganensissubsp.michiganensisOn Rockwool-Grown Greenhouse TomatoesTwo greenhouse experiments were conducted in which tomato plants artificially inoculated withClavibacter michiganensissubsp.michiganensis(Cmm) were grown in an open rockwool system as spring and autumn crops. Two isolates of the rhizosphere bacteria,Pseudomonas fluorescensstrain PSR21,Pseudomonas reactansstrain GGS14, a commercial biocontrol agent Aqua Bac Plus (Bacillusspp.) and a proprietary disinfectant containing QAC+Chx, applied at weekly intervals, were evaluated for their efficiency in the suppression of the bacterial canker of tomato. All treatments tested revealed to be ineffective in controlling the disease. The introduction ofCmmbacteria into the fresh rockwool in the first year of its usage resulted in a 100% death of tomato plants, whereas following an artificial inoculation of two- and three-year-old rockwool slabs withCmmbacteria dead plants amounted to 70 and 58%, respectively. This indicates that in the re-used rockwool a natural microbial suppressiveness to bacterial canker of tomato might be developed in the root zone.

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“…SCM, which was developed for the closely related C. michiganensis subsp, michiganensis (Fatmi and Schaad, 1988), showed complete inhibition of Cms colonies in the pour platings of this semi-selective medium. Subsequent experiments (Table 2) showed the inhibiting effect of nalidixic acid at 10 rag/1 on colonies of Cms, providing a possible explanation for the complete inhibition of Cms on SCM which contained 30 rag/1.…”

Section: Selective Cms Medium For Pour Platingmentioning

confidence: 95%

Development of a semi-selective medium and an immunofluorescence colony-staining procedure for the detection of Clavibacter michiganensis subsp. sepedonicus in cattle manure slurry

Roozen¹,

Vuurde²

1991

Netherlands Journal of Plant Pathology

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Various compounds and basal media were tested for their suitability to create a semi-selective medium for isolation of Clavibaeter michiganensis subsp, sepedonicus (Cms) from cattle manure slurry containing c. 108 colony forming units (cfu) per ml.Plating efficiency of Cms in yeast glucose mineral medium (YGM) was 104% compared with yeast peptone glucose medium. Nalidixic acid, polymyxin B sulphate and the experimental disinfectant S-0208 inhibited colony growth of cattle slurry bacteria as compared with Cms in YGM. The optimal concentration of these inhibitors in combination was determined by modified agar diffusion tests and by pour plating in 24-well tissue culture plates. The semi-selective medium YGM1 consisted of YGM supplemented with nalidixic acid (2 rag/l), polymyxin B sulphate (30 rag/l) and S-0208 (125 rag/l). Plating efficiency varied for Cms between 50.9 and 69.6%, for cattle slurry bacteria between 1.8 and 2.5% and for saprophytes from potato heel end extracts between 11.5 and 27.4%.Differentiation of Cms colonies from other colonies was based on their small and bluish colony morphology in pour plates and on immunofluorescence colony-staining (IFC). IFC of a pure culture of micro colonies of Cms in YGM was possible after one day incubation (colonies c. 5 cells). Green background fluorescence in the agar gels was prevented by addition of Tween 20 (0.1%) to the washing buffer and the use of 1% agar gels. IFC of macro colonies of Cms in YGMI, visible with 4 x objective magnification, was possible after 4 days. The detection level of the target organism in artificially inoculated cattle slurry in YGMI based on colony morphology varied between 1.4 x 103 and 2.3 x 10 4 cfu per ml of cattle slurry. Miniaturized plating combined with IFC, using wells in tissue culture plates (@ --16 ram), proved suitable for detection, but was c. 30 times less sensitive. The recovery of Cms was negatively correlated with the number of saprophytic colonies in the agar plates (R 2 = 0.74).

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Semiselective Agar Medium for Isolation ofClavibacter michiganensesubsp.michiganensefrom Tomato Seed

Cited by 59 publications

References 0 publications

Evidence for plasmid-encoded virulence factors in the phytopathogenic bacterium Clavibacter michiganensis subsp. michiganensis NCPPB382

Evidence for plasmid-encoded virulence factors in the phytopathogenic bacterium Clavibacter michiganensis subsp. michiganensis NCPPB382

Attempts at Biological Control of Clavibacter michiganensis subsp. michiganensis On Rockwool-Grown Greenhouse Tomatoes

Development of a semi-selective medium and an immunofluorescence colony-staining procedure for the detection of Clavibacter michiganensis subsp. sepedonicus in cattle manure slurry

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