Modified crystal violet pectate medium (CVP) based on a new polypectate source (Slendid) for the detection and isolation of soft rot erwinias

Hyman, L. J.; Sullivan, L.; Toth, Ian K.; Pérombelon, M. C. M.

doi:10.1007/bf02357904

Cited by 49 publications

(30 citation statements)

References 3 publications

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“…After 4-5 days, the colony was appeared resemble a fried egg, with a blue); the result is compatible with [7] and [10] and which were oxidase-negative, this reaction was observed later than 60 seconds at 25-30° C (purple coloration) which means the reaction was negative and the isolate were Erwinia spp., the result was compatible with [10], catalasepositive by formation of the gas bubbles, which indicated a positive reaction, that would mean the isolates were as Erwinia spp., the result was compatible with [10] and consisted of Gram-negative, non-motile rods were picked after cultivation at 30 °C for 24 hs. ; the result is compatible with [13].…”

Section: Isolation and Initial Characterizationsupporting

confidence: 77%

“…; the result is compatible with [13]. After the purification procedures, all erwinias isolates were screened for fermentation of suger using Sorbitol, Raffinose and Cellobiose discs then the twenty local erwinias isolates EM (1)(2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14)(15)(16)(17)(18)(19)(20) were selected on the base of this screening; (5 from Awrad Alnahar potato's store, 4 from Alradwaniyah potato's store and 11 from Albyaa Market) that was according to [10]/ (HiMedia Laboratories).…”

Section: Isolation and Initial Characterizationmentioning

confidence: 64%

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Detection of local Erwinia Isolates Causing Diseases in Potato by Using DNA Amplification by Polymerase Chain Reaction Technique (PCR)

Mohammed

Selman

2013

JNUS

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Seventy samples of infected potato's tubers were collected from Baghdad city during the period from September 2011 to February 2012. These samples were collected from Alradwaniyah potato's store/ Albyaa market and from Awrad Alnahar potato's store in Abu ghraib and two bacterial strains of Erwinia were obtained from Plant Protection department/ College of Agriculture/ Baghdad University. The Crystal Violet Pectate (CVP) used as a selective-diagnostic medium to isolate and purify Erwinia. The strains and isolates of Erwinia were characterized according to the morphology and microscopic characteristics, along with the biochemical tests and API 20E tests. The Erwinia isolates have a positive reaction for Catalase, Raffinose and Cellobiose and a negative reaction for Oxidase and Sorbitol. By using Reagent Genomic DNA Kit method for Total DNA extraction, during electrophoresis process, the results showed that some of selected isolates and the two standard strains of Erwinia contained at least two bands of small plasmid DNA. The identification was confirmed by using DNA amplification by polymerase chain reaction technique (PCR) and Erwinia carotovora subsp. carotovora specific primer such as (Ec001), the selected local isolates of Erwinia and Erwinia carotovora subsp. carotovora ATCC 15713 potato's pathogen had a positive reaction (one band 312 bp) for the former primer.

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Section: Isolation and Initial Characterizationsupporting

confidence: 77%

Section: Isolation and Initial Characterizationmentioning

confidence: 64%

Detection of local Erwinia Isolates Causing Diseases in Potato by Using DNA Amplification by Polymerase Chain Reaction Technique (PCR)

Mohammed

Selman

2013

JNUS

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“…brasiliense by plating on CVP medium and IGS-specific PCR with the primers BR1f/L1r, which are species specific for P. carotovorum subsp. brasiliense (18,24). These pMP7604 plasmid-carrying P. carotovorum subsp.…”

Section: Discussionmentioning

confidence: 99%

Colonization Patterns of an mCherry-Tagged Pectobacterium carotovorum subsp. brasiliense Strain in Potato Plants

Kubheka¹,

Coutinho²,

Moleleki³

et al. 2013

Phytopathology®

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Pectobacterium carotovorum subsp. brasiliense is a newly identified member of the potato soft rot enterobacteriaceae. The pathogenesis of this pathogen is still poorly understood. In this study, an mCherry-P. carotovorum subsp. brasiliense-tagged strain was generated to study P. carotovorum subsp. brasiliense-potato plant interactions. Prior to use, the tagged strain was evaluated for in vitro growth, plasmid stability, and virulence on potato tubers and shown to be similar to the wild type. Four potato cultivars were evaluated for stem-based resistance against P. carotovorum subsp. brasiliense. Confocal laser-scanning microscopy and in vitro viable cell counts showed that P. carotovorum subsp. brasiliense is able to penetrate roots of a susceptible potato cultivar as early as 12 h postinoculation and migrate upward into aerial stem parts. Due to the phenotypic differences observed between tolerant and susceptible cultivars, a comparison of P. carotovorum subsp. brasiliense colonization patterns in these cultivars was undertaken. In the susceptible cultivar, P. carotovorum subsp. brasiliense cells colonized the xylem tissue, forming "biofilm-like" aggregates that led to occlusion of some of the vessels. In contrast, in the tolerant cultivar, P. carotovorum subsp. brasiliense appeared as free-swimming planktonic cells with no specific tissue localization. This suggests that there are resistance mechanisms in the tolerant cultivar that limit aggregation of P. carotovorum subsp. brasiliense in planta and, hence, the lack of symptom development in this cultivar.Additional keyword: Dickeya.

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“…The bacteria were isolated by the methods described by Schaad et al (29) and Yap et al (35). Briefly, bacteria from decayed tubers were streaked either onto crystal violet pectate (CVP) medium prepared with either Bulmer or Slendid pectate (16) or onto a raffinose (RAF) medium modified from the medium described by Segall (30), and the cultures were incubated at room temperature for at least 3 days. The modified RAF medium consists of (per liter) 10 g of raffinose, 2 g of K 2 HPO 4 , 5 g of ammonium sulfate, 0.4 g of eosin Y, 0.065 g of methylene blue, and 18 g of agar.…”

Section: Methodsmentioning

confidence: 99%

Phylogeny and Virulence of Naturally Occurring Type III Secretion System-Deficient Pectobacterium Strains

Kim¹,

Perna

et al. 2009

Appl Environ Microbiol

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Pectobacterium species are enterobacterial plant-pathogenic bacteria that cause soft rot disease in diverse plant species. Previous epidemiological studies of Pectobacterium species have suffered from an inability to identify most isolates to the species or subspecies level. We used three previously described DNA-based methods, 16S-23S intergenic transcribed spacer PCR-restriction fragment length polymorphism analysis, multilocus sequence analysis (MLSA), and pulsed-field gel electrophoresis, to examine isolates from diseased stems and tubers and found that MLSA provided the most reliable classification of isolates. We found that strains belonging to at least two Pectobacterium clades were present in each field examined, although representatives of only three of five Pectobacterium clades were isolated. Hypersensitive response and DNA hybridization assays revealed that strains of both Pectobacterium carotovorum and Pectobacterium wasabiae lack a type III secretion system (T3SS). Two of the T3SS-deficient strains assayed lack genes adjacent to the T3SS gene cluster, suggesting that multiple deletions occurred in Pectobacterium strains in this locus, and all strains appear to have only six rRNA operons instead of the seven operons typically found in Pectobacterium strains. The virulence of most of the T3SS-deficient strains was similar to that of T3SS-encoding strains in stems and tubers.The genus Pectobacterium (formerly Erwinia) contains both narrow-and broad-host-range bacterial plant pathogens that cause soft rot, stem rot, wilt, and blackleg in species belonging to over 35% of plant orders (20). Four Pectobacterium species have been described: Pectobacterium atrosepticum, Pectobacterium betavasculorum, Pectobacterium carotovorum, and Pectobacterium wasabiae (9). The recently described organism P. carotovorum subsp. brasiliensis is genetically distinct from previously described Pectobacterium taxa; approximately 82% of its genes are shared with P. atrosepticum, and 84% of its genes are shared with P. carotovorum subsp. carotovorum, while 13% of its genes are found in neither P. atrosepticum nor P. carotovorum subsp. carotovorum (7,10,20). To date, only P. carotovorum subsp. carotovorum and P. atrosepticum have been reported to occur in the same field (14, 21). P. carotovorum subsp. carotovorum is found worldwide, and P. atrosepticum is found in cool climates; while P. carotovorum subsp. brasiliensis has been found only in Brazil, Israel, and the United States, it is likely to have a wider distribution (20). Compared to the ecology and genetics of P. carotovorum subsp. carotovorum and P. atrosepticum, little is known about the ecology and genetics of P. betavasculorum, P. wasabiae, or P. carotovorum subsp. brasiliensis.Pectobacterium strains isolated from potato are diverse based on serology, genome structure, and fatty acid composition (5, 35). Previous epidemiological studies of pectolytic Enterobacteriaceae were complicated by the diversity of this group and the lack of tools capable of placing all isolates ...

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Modified crystal violet pectate medium (CVP) based on a new polypectate source (Slendid) for the detection and isolation of soft rot erwinias

Cited by 49 publications

References 3 publications

Detection of local Erwinia Isolates Causing Diseases in Potato by Using DNA Amplification by Polymerase Chain Reaction Technique (PCR)

Detection of local Erwinia Isolates Causing Diseases in Potato by Using DNA Amplification by Polymerase Chain Reaction Technique (PCR)

Colonization Patterns of an mCherry-Tagged Pectobacterium carotovorum subsp. brasiliense Strain in Potato Plants

Phylogeny and Virulence of Naturally Occurring Type III Secretion System-Deficient Pectobacterium Strains

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