An Improved Agar Plating Assay for DetectingPseudomonas syringaepv.syringaeandP. s.pv.phaseolicolain Contaminated Bean Seed

Mohan, S. K.; Schaad, N. W.

doi:10.1094/phyto-77-1390

Cited by 113 publications

(61 citation statements)

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“…The overall objective of this study was to evaluate the detection method and determine the occurrence of Psp on common bean seed collected from different genotypes and populations in Serbia. A number of plating assays and semi-selective media have been developed and improved to detect Psp in bean seed (Mohan and Schaad, 1987;Gozczynska and Serfontein, 1998). We conducted a detection method by sowing the extract obtained from the soaked whole bean seed onto the semi-selective media MSP and MT.…”

Section: Discussionmentioning

confidence: 99%

“…This study confirmed the advantage of the MT medium, which allowed simultaneous detection of Xap and Psp on the tested bean cultivar Oplenac, where seeds were infected with both of the bacteria. Other media such as King B (Taylor, 1970), KBC (Mohan and Schaad, 1987;NSHS, 2002) and LPGA can also be used for the isolation of Psp from bean seeds.…”

Section: Discussionmentioning

confidence: 99%

“…After incubating, 0.1 mL of undiluted extracts obtained from the seeds and their 10-fold dilution series (to 10 -5 ) were plated onto the surface of two semi-selective media, Modified Sucrose Peptone Agar (MSP) and Milk Tween Agar (MT), described by Mohan and Schaad (1987) and Goszczynska and Serfontein (1998), respectively. The series of dilution were prepared with the aim to get single colonies of Psp which could be morfologically differentiated from other non-targeted organisms.…”

Section: Isolation On Semi-selective Mediamentioning

confidence: 99%

“…The detection of bacterium on seeds is essential for effective control of the disease. Several methods have been described for testing the presence of Psp in bean seed: a seed soakplant inoculation technique (Lahman and Schaad, 1985;Webster et al, 1983), immunological methods (Guthrie et al, 1965;Van Vuurde et al, 1983), plating on King B agar (Van Vuurde and Van den Bovenkamp, 1987) and semiselective modified sucrose peptone (MSP) agar (Mohan and Schaad, 1987). Various PCR assays have been proposed for the detection of Psp (Prosen et al, 1993;Schaad et al, 1995Schaad et al, , 2001Mosqueda and Herrera, 1997).…”

Section: Introductionmentioning

confidence: 99%

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Application of semi-selective mediums in routine diagnostic testing of Pseudomonas savastanoi pv. phaseolicola on common bean seeds

Popović

Milovanović²,

Aleksić

et al. 2012

Sci. agric. (Piracicaba, Braz.)

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Halo blight, caused by Pseudomonas savastanoi pv. phaseolicola (Psp), is considered to be an important bacterial disease on common bean (Phaseolus vulgaris L.) in Serbia. Use of pathogen-free seeds is one of the most effective control measures against this disease. The aim of this study was to evaluate a detection method for Psp on untreated common bean seeds (23 genotypes) from commercial crops grown within Serbia. Detection of this pathogen was made by plating onto the modified sucrose peptone (MSP) and Milk Tween (MT) semi-selective mediums from soaked whole common bean seed. Colonies growing on the MSP medium were light yellow, convex and shiny, whereas on the MT medium, they were creamy white, flat and circular. The pathogenicity of the obtained strains was confirmed by the inoculation of germinated bean seed. The isolates recovered from the seed assay were further confirmed to be Psp by using both Enzyme-linked immunosorbent assay (ELISA) and Polymerase Chain Reaction (Nested-PCR) detection methodologies. The International Seed Testing Association (ISTA) method selected for this work was found to be effective in detecting the presence of Psp in common bean seed. The bacterium Psp was detected in only two of the 23 seed samples analyzed by this method, which shows that the bacterium is not widespread in Serbia.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Isolation On Semi-selective Mediamentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Application of semi-selective mediums in routine diagnostic testing of Pseudomonas savastanoi pv. phaseolicola on common bean seeds

Popović

Milovanović²,

Aleksić

et al. 2012

Sci. agric. (Piracicaba, Braz.)

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show abstract

“…At each sampling time for up to 7 days after inoculation, 3-6 plants were randomly sampled from the 12 plants inoculated with each strain, weighed, and individually macerated in 10 ml of 0.1 M phosphate buffer. Aliquots of the macerate were diluted and plated on KBC medium (Mohan and Schaad, 1987), the same medium used to isolate these strains from the environment. Colonies were counted after 4 days of incubation at 22-25 1C in the dark.…”

Section: Methodsmentioning

confidence: 99%

Pseudomonas syringae naturally lacking the canonical type III secretion system are ubiquitous in nonagricultural habitats, are phylogenetically diverse and can be pathogenic

et al. 2012

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The type III secretion system (T3SS) is an important virulence factor of pathogenic bacteria, but the natural occurrence of variants of bacterial plant pathogens with deficiencies in their T3SS raises questions about the significance of the T3SS for fitness. Previous work on T3SS-deficient plant pathogenic bacteria has focused on strains from plants or plant debris. Here we have characterized T3SS-deficient strains of Pseudomonas syringae from plant and nonplant substrates in pristine nonagricultural contexts, many of which represent recently described clades not yet found associated with crop plants. Strains incapable of inducing a hypersensitive reaction (HR À ) in tobacco were detected in 65% of 126 samples from headwaters of rivers (mountain creeks and lakes), snowpack, epilithic biofilms, wild plants and leaf litter and constituted 2 to 100% of the P. syringae population associated with each sample. All HR À strains lacked at least one gene in the canonical hrp/hrc locus or the associated conserved effector locus, but most lacked all six of the genes tested (hrcC, hrpL, hrpK1, avrE1 and hrpW1) and represented several disparate phylogenetic clades. Although most HR À strains were incapable of causing symptoms on cantaloupe seedlings as expected, strains in the recently described TA-002 clade caused severe symptoms in spite of the absence of any of the six conserved genes of the canonical T3SS according to PCR and Southern blot assays. The phylogenetic context of the T3SS variants we observed provides insight into the evolutionary history of P. syringae as a pathogen and as an environmental saprophyte.

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Detection and Identification of Bacterial and Phytoplasmal Pathogens

2017

Microbial Plant Pathogens

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An Improved Agar Plating Assay for DetectingPseudomonas syringaepv.syringaeandP. s.pv.phaseolicolain Contaminated Bean Seed

Cited by 113 publications

References 0 publications

Application of semi-selective mediums in routine diagnostic testing of Pseudomonas savastanoi pv. phaseolicola on common bean seeds

Application of semi-selective mediums in routine diagnostic testing of Pseudomonas savastanoi pv. phaseolicola on common bean seeds

Pseudomonas syringae naturally lacking the canonical type III secretion system are ubiquitous in nonagricultural habitats, are phylogenetically diverse and can be pathogenic

Detection and Identification of Bacterial and Phytoplasmal Pathogens

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