Development of a selective medium and antisera for Pseudomonas syringae pv. syringae from seeds of barley and wheat

The article presents results of studies on development of isolation scheme and bacteriological identification of Pseudomonas syringae bacteria and its approbation. The introduction of the article describes the objects of Pseudomonas syringae contamination - fruit trees and shrubs, agricultural plants, which proves the relevance of the research in the field of expanding of laboratory methods for identifying phytopathogenic microorganisms. The author’s bacteriological scheme includes the use of King B Medium (Pseudomonas F Agar; Pronadisa 1532) as a selective medium. Initially, the isolated bacteria are differentiated to Pseudomonas genus, the following paramters are studied: anaerobic fermentation, production of enzymes catalase, lecithinase, lipase; hydrolysis of starch and gelatin; fermentation of glucose and lactose, also, a test for maceration is put. The second stage of the research includes the study of the growth of bacterial cultures on meat-and-peptone agar at 41 ° C and at 5% of NaCl; oxidase production, arginine hydrolase; fermentation of mannose and sorbitol; formation of levan, hydrogen sulfide and indole, esculin, a hypersensitivity reaction is set. The determined parametres allow to type the representatives of the genus to Pseudomonas syringae species within 192 hours. During the research, a collection of 12 strains of Pseudomonas syringae bacteria was formed, isolated from 97 objects of phytosanitary supervision and identified according to the developed technique. The proposed bacteriological scheme allows to differentiate the above microorganisms on the basis of the analysis of 25 parameters. The application of a phage biological product as a diagnosticum (according to the Otto method) expands the spectrum of the analyzed biological properties of the isolated and identified Pseudomonas syringae bacteria.

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Highly sensitive detection of Pseudomonas syringae pv. japonica from wheat seeds by selective medium and membrane filter method

Annual Report of The Kansai Plant Protection Society ◽

10.4165/kapps.56.43 ◽

2014 ◽

Vol 56 (0) ◽

pp. 43-47

Author(s):

Hirofumi Suzuki ◽

Katsutoshi Kuroda ◽

Tomohiro Yamakawa ◽

Kengo Matsumoto ◽

Fujio Hashizume ◽

...

Keyword(s):

Pseudomonas Syringae ◽

Membrane Filter ◽

Selective Medium ◽

Sensitive Detection ◽

Filter Method ◽

Highly Sensitive ◽

Membrane Filter Method ◽

Highly Sensitive Detection ◽

Wheat Seeds

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A Selective Medium for Isolation of Pseudomonas syringae, the Pathogen of Bacterial Blossom Blight of Kiwifruit.

Japanese Journal of Phytopathology ◽

10.3186/jjphytopath.60.729 ◽

1994 ◽

Vol 60 (6) ◽

pp. 729-734 ◽

Cited By ~ 1

Author(s):

Takanori MIYOSHI ◽

Yasunobu TACHIBANA

Keyword(s):

Pseudomonas Syringae ◽

Selective Medium

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Loop-mediated isothermal amplification: a rapid molecular technique for early diagnosis of Pseudomonas syringae pv. syringae of stone fruits

Journal of Genetic Engineering and Biotechnology ◽

10.1186/s43141-020-00062-6 ◽

2020 ◽

Vol 18 (1) ◽

Author(s):

R. Goudarzi ◽

M. M. Mortazavi

Keyword(s):

Pseudomonas Syringae ◽

Pathogenic Bacteria ◽

Selective Medium ◽

Isothermal Amplification ◽

Plant Diseases ◽

Molecular Technique ◽

Specific Method ◽

Stone Fruits ◽

Serological Tests ◽

Loop Mediated Isothermal Amplification

Abstract Background Pathogenic bacteria cause significant economic damages in agriculture. The detection of such bacteria is considered as a continual interest for plant pathologists to prevent disease dissemination. Pseudomonas syringae pv. syringae is one of the most important bacterial pathogens infecting yield and quality of stone fruits throughout the world. Biochemical assays such as a LOPAT and GATTa are common methods to detect this pathogen. Serological tests and culturing on King’s B selective medium also used to isolate this bacterium. Selective media is composed of specific and effective ingredients to inhibit the growth of certain species of microbes in a mixed culture while allowing others to grow. These are used for the growth of only selected microorganisms. King’s B medium can be used as a general medium for the non-selective isolation cultivation and pigment production of Pseudomonas species from foods, cosmetic samples, plants, etc. Nevertheless, the mentioned methods are not enough accurate to differentiate the strains. On the other hand, PCR-based techniques are sensitive and efficient in detecting plant diseases. However, these techniques are not practicable for those researchers who do not have access to a thermal cycler. We have used loop-mediated isothermal amplification to couple with a target. The amplification of syrD gene using loop and bumper primers can be used to prevent disease dissemination. Results The outcome of this investigation indicated more sensitivity of LAMP in comparison to PCR. The direct addition of SYBR Gold in microtube is more sensitive than gel in both LAMP and PCR byproducts so we can eliminate gel electrophoresis, while the LAMP showed high sensitivity and high specificity in comparison to results obtained by cultivation. The described molecular test could detect Pseudomonas syringae pv. syringae type in nearly 1 h, and this is the first time that Lamp molecular detection of Pseudomonas syringae pv. syringae particularly on stone fruits is described and introduced. Conclusions The obtained data confirmed that LAMP is a fast, cheap, and high specific method for the rapid detection of Pseudomonas syringae pv. syringae to the comparison of PCR and culture.

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Identification of Erwinia amylovora by Growth Morphology on Agar Containing Copper Sulfate and by Capsule Staining with Lectin

Plant Disease ◽

10.1094/pdis.1998.82.2.158 ◽

1998 ◽

Vol 82 (2) ◽

pp. 158-164 ◽

Cited By ~ 53

Author(s):

Stefan Bereswill ◽

Susanne Jock ◽

Peter Bellemann ◽

Klaus Geider

Keyword(s):

Pseudomonas Syringae ◽

Copper Sulfate ◽

Fire Blight ◽

Erwinia Amylovora ◽

Selective Medium ◽

Colony Morphology ◽

Associated Bacteria ◽

Yellow Color ◽

Pseudomonas Syringae Pathovars ◽

Fluorescent Compounds

Erwinia amylovora strains formed yellow colonies on minimal agar medium MM2 containing asparagine and copper sulfate (MM2Cu), in contrast to a white morphology on minimal agar without copper salt. Additionally, the colonies were mucoid to various extents. The yellow color was characteristic for the fire blight pathogen, including strains from raspberry and from other unusual host plants, and was used to establish a novel plating technique for identification of E. amylovora. The new identification method was especially superior to semi-selective media with sucrose when natural levan-deficient strains were assayed. No growth of E. amylovora was observed for the similar medium MM1 containing 2 mM CuSO4, due to its low content of as paragine. Identification by colony morphology on MM2 agar with copper was confirmed by staining the bacterial capsules with FITC-labeled lectin from Abrus precatorious, a compound which has a high affinity for galactose residues, the main sugar in the capsular exopolysaccharide amylovoran of E. amylovora. Other plant-associated bacteria usually did not produce the typical colony morphology of E. amylovora on MM2 agar with copper. Furthermore, those cells were not stained with the Abrus lectin. Capsule staining was also observed for weakly mucoid strains of E. amylovora, but not for strains with mutations affecting amylovoran synthesis. The secretion of fluorescent compounds by Pseudomonas syringae pathovars and even growth of any other bacterial colonies adjacent to E. amylovora could interfere with the formation of typical yellow colonies on MM2Cu, which could be white in case of dense plating. After screening for white colonies on LB agar, E. amylovora was identified in extracts from Cotoneaster leaves and in bark from apple trees with fire blight symptoms by its yellow growth pattern on MM2Cu agar and by capsule staining. The proposed selective medium gives a clear signal, is easy to prepare, does not contain dyes or any compounds toxic to humans, and can also detect E. amylovora strains deficient in levan synthesis.

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Effects of the pseudomonad phytotoxin coronatine on tomato leaf structure and ultrastructure

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100140683 ◽

1995 ◽

Vol 53 ◽

pp. 862-863

Author(s):

D.A. Palmer ◽

C.L. Bender

Keyword(s):

Pseudomonas Syringae ◽

Membrane Integrity ◽

Leaf Tissue ◽

Cell Number ◽

Cell Ultrastructure ◽

Response To Treatment ◽

Prominent Symptom ◽

Chlorophylls A And B ◽

Cell Dimensions ◽

Structure And Ultrastructure

Coronatine is a non-host-specific phytotoxin produced by several members of the Pseudomonas syringae group of pathovars. The toxin acts as a virulence factor in P. syringae pv. tomato, allowing the organism to multiply to a higher population density and develop larger lesions than mutant strains unable to produce the toxin. The most prominent symptom observed in leaf tissue treated with coronatine is an intense spreading chlorosis; this has been attributed to a loss of chlorophylls a and b in tobacco. Coronatine's effects on membrane integrity and cell ultrastructure have not been previously investigated. The present study describes changes in tomato leaves in response to treatment with purified coronatine, infection by a coronatine-producing strain of P. syringae pv. tomato, and infection by a cor" mutant.In contrast to H2O-treated tissue, coronatine-treated tissue showed a diffuse chlorosis extending approximately 5 mm from the inoculation site. Leaf thickness, cell number, and cell dimensions were similar for both healthy and coronatine-treated, chlorotic tissue; however, the epidermal cell walls were consistently thicker in coronatine-treated leaves (Figs, la and lb).

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