Quantitative real-time PCR assay for rapid detection of Pseudomonas amygdali pv. lachrymans in cucumber leaf rinse

Gazdík, Filip; Peňázová, Eliška; Čechová, Jana; Baránek, Miroslav; Eichmeier, Aleš

doi:10.1007/s41348-019-00258-0

Cited by 3 publications

(2 citation statements)

References 46 publications

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“…The Psl was identified in samples by qPCR according to Gazdik et al [27] and based on a sequence of the nfrB gene. A 2 µL of sample was added to reaction mixture consisting of 5 µL 2×SensiFast Sybr No-Rox Mix, 1 µL of each primer (10 µM): Pal-nfrB-fwd (5 -GTA CGG TAT GCA AGC GAT CTTC-3 ) and Pal-nfrB-rev (5 -CCG AAA CAG GTA ACG GT-3 ) [27], and 1 µL of water. Quantification of bacteria was based on a standard curve made of three-fold dilutions of genomic DNA of the reference strain Psl 1729 and is presented in Figure 1.…”

Section: Bacterial Samples Verification and Quantificationmentioning

confidence: 99%

“…Table 4 presents in detail the comparison of various detection methods of Pseudomonas syringae species mentioned in this article with the proposed electrochemical biosensor. According to the standard PCR method, which detects bacterial cells, this assay is less sensitive than real-time qPCR [27]. The real-time qPCR LOD is 1.3 times lower, but these values are very similar, in the same order of magnitude.…”

Section: Psl Bacteria Detection On Developed Electrochemical Immunosementioning

confidence: 99%

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Detection of the Plant Pathogen Pseudomonas Syringae pv. Lachrymans on Antibody-Modified Gold Electrodes by Electrochemical Impedance Spectroscopy

Cebula¹,

Żołędowska²,

Dziąbowska³

et al. 2019

Sensors

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The present work describes an impedimetric immunosensor for Pseudomonas syringae pv. lachrymans (Psl) detection. This pathogen infects many crop species causing considerable yield losses, thus fast and cheap detection method is in high demand. In the assay, the gold disc electrode was modified with 4-aminothiophenol (4-ATP), glutaraldehyde (GA), and anti-Psl antibodies, and free-sites were blocked with bovine serum albumin (BSA). Sensor development was characterized by cyclic voltammetry (CV) and antigen detection by electrochemical impedance spectroscopy (EIS) measurements. Seven analyzed strains of Psl were verified as positive by the reference method (PCR) and this immunoassay, proving sensor specificity. Label-free electrochemical detection was in the linear range 1 × 103–1.2 × 105 CFU/mL (colony-forming unit) with an R2 coefficient of 0.992 and a detection limit (LOD) of 337 CFU/mL. The sensor did not interfere with negative probes like buffers and other bacteria. The assay was proven to be fast (10 min detection) and easy in preparation. The advantage was the simplicity and availability of the verified analyte (whole bacteria) as the method does not require sample pretreatment (e.g., DNA isolation). EIS biosensing technique was chosen as one of the simplest and most sensitive with the least destructive influence on the probes compared to other electrochemical methods.

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Section: Bacterial Samples Verification and Quantificationmentioning

confidence: 99%

Section: Psl Bacteria Detection On Developed Electrochemical Immunosementioning

confidence: 99%

Detection of the Plant Pathogen Pseudomonas Syringae pv. Lachrymans on Antibody-Modified Gold Electrodes by Electrochemical Impedance Spectroscopy

Cebula¹,

Żołędowska²,

Dziąbowska³

et al. 2019

Sensors

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Guttation Fluids Containing Pseudomonas amygdali pv. lachrymans Are a Potential Source of Secondary Infection in Cucumber

Zhang

Wang

et al. 2021

Phytopathology®

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Guttation is a common feature of cucumber leaves under high relative humidity conditions; however, little is known about the role of guttation in the transmission of Pseudomonas amygdali pv. lachrymans (Pal), which is the pathogen of cucumber angular leaf spot (ALS) disease. In this study, experimental evidence for the transmission of Pal inside cucumber plants and through guttation was provided, and the results proved that Pal can be transmitted from the bottom leaf to the upper leaves inside the plant and excreted from the upper leaves through guttation. After that, the third leaf of cucumber was inoculated with Pal bacterial suspension, Pal was detected on the fifth leaf, the petiole, and the stem and in guttation drops. Healthy cucumber seedlings were infected by Pal in the guttation droplets, indicating that guttation fluids containing Pal could become a potential source of secondary infection. The results from this study verified the hypothesis that guttation is a potential route for Pal excretion from cucumber plants and may be a source of secondary transmission under high relative humidity conditions.

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Applicability of Smart Tools in Vegetable Disease Diagnostics

Ovesná

Tsiropoulos²,

Collier

et al. 2023

Agronomy

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Various diseases and pests cause serious damage to vegetable crops during the growing season and after harvesting. Growers attempt to minimize losses by protecting their crops, starting with seed and seedling treatments and followed by monitoring their stands. In many cases, synthetic pesticide treatments are applied. Integrated pest management is currently being employed to minimize the impact of pesticides upon human health and the environment. Over the last few years, “smart” approaches have been developed and adopted in practice to predict, detect, and quantify phytopathogen occurrence and contamination. Our review assesses the currently available ready-to-use tools and methodologies that operate via visual estimation, the detection of proteins and DNA/RNA sequences, and the utilization of brand-new innovative approaches, highlighting the availability of solutions that can be used by growers during the process of diagnosing pathogens.

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Quantitative real-time PCR assay for rapid detection of Pseudomonas amygdali pv. lachrymans in cucumber leaf rinse

Cited by 3 publications

References 46 publications

Detection of the Plant Pathogen Pseudomonas Syringae pv. Lachrymans on Antibody-Modified Gold Electrodes by Electrochemical Impedance Spectroscopy

Detection of the Plant Pathogen Pseudomonas Syringae pv. Lachrymans on Antibody-Modified Gold Electrodes by Electrochemical Impedance Spectroscopy

Guttation Fluids Containing Pseudomonas amygdali pv. lachrymans Are a Potential Source of Secondary Infection in Cucumber

Applicability of Smart Tools in Vegetable Disease Diagnostics

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