Detection and Quantification of <i>Peronospora tabacina</i> Using a Real-Time Polymerase Chain Reaction Assay

Blanco‐Meneses, Monica; Ristaino, Jean Beagle

doi:10.1094/pdis-05-10-0333

Cited by 19 publications

(13 citation statements)

References 33 publications

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“…Quantitative PCR can also be a valuable tool in the selection of resistant species and/or cultivars, because molecular data can be detected earlier, enabling the selection of resistant plants even when samples are indistinguishable based on visual assessment (Blanco‐Meneses and Ristaino ; Montes‐Borrego et al. ).…”

Section: Detection Of Phytopathogenic Fungi and Oomycetes In Host Tismentioning

confidence: 99%

“…). Due to its high specificity and sensitivity, qPCR is increasingly included in official protocols of the European Plant Protection Organization (http://archives.eppo.org/index.htm) for the certification, production and assessment of healthy plant materials (Blanco‐Meneses and Ristaino ; Boutigny et al. ).…”

Section: Detection Of Phytopathogenic Fungi and Oomycetes In Host Tismentioning

confidence: 99%

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Use of QuantitativePCRDetection Methods to Study Biocontrol Agents and Phytopathogenic Fungi and Oomycetes in Environmental Samples

Sanzani

Nicosia

Faedda³

et al. 2013

Journal of Phytopathology

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Quantitative polymerase chain reaction (qPCR) is a versatile technique for the accurate, sensitive, reliable and high‐throughput detection and quantification of target DNA in various environmental samples, and in recent years, it has greatly contributed to the advancement of knowledge in the plant pathology field. Indeed, this technique is ideal to evaluate inoculum threshold levels and to study the epidemiology, biology and ecology of phytopathogenic fungi and oomycetes, thus opening up new research opportunities to investigate host–pathogen interactions and to address tasks related to quarantine, eradication and biosecurity. Moreover, it can be a useful tool in breeding programs. The present review analyses the most relevant applications of qPCR for the detection and quantification of filamentous fungi and oomycetes within host tissues and in soil, air and water, along with brief paragraphs focusing on new application fields such as the detection and quantification of mycotoxigenic fungi and biocontrol agents. The high potentiality of qPCR for present and future applications is highlighted together with a critical analysis of major drawbacks that need to be corrected to definitively confirm it as a preferential routine quantitative detection method.

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Section: Detection Of Phytopathogenic Fungi and Oomycetes In Host Tismentioning

confidence: 99%

Section: Detection Of Phytopathogenic Fungi and Oomycetes In Host Tismentioning

confidence: 99%

Use of QuantitativePCRDetection Methods to Study Biocontrol Agents and Phytopathogenic Fungi and Oomycetes in Environmental Samples

Sanzani

Nicosia

Faedda³

et al. 2013

Journal of Phytopathology

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“…Although there are numerous studies describing qPCR to detect and quantify microorganisms from environmental samples (7,9,26,36,42), only a few studies relate the quantity of inoculum detected using qPCR with subsequent disease to determine a disease risk threshold (8,16). The results from this assay indicate that ascospore counts determined using the qPCR assay are related to subsequent disease.…”

Section: Discussionmentioning

confidence: 99%

“…Quantitative polymerase chain reaction (qPCR)-based methods have been developed to specifically detect and quantify microorganisms from air, plant, or soil samples (7,9,16,38,42,49,55,59). The advantages of qPCR are its high sensitivity and specificity.…”

mentioning

confidence: 99%

Evaluation of Air Sampling and Detection Methods to Quantify Airborne Ascospores of Sclerotinia sclerotiorum

Parker¹,

McDonald²,

Boland

2014

Plant Disease

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Parker, M. L., McDonald, M. R., and Boland, G. J. 2014. Fvaluation of air sampling and detection methods to quantify airborne ascospores of Scierotinia scierotiorum. Plant Dis. 98:32-42.Detection and quantification of airborne ascospores as a component of the Sclerotinia rot of carrot (SRC) forecast model is currently accomphshed using the blue plate test (BPT), which uses Sclerotinia semiselective medium (SSM). A quantitative polymerase chain reaction (qPCR) assay was developed to reduce the time to specifically quantify ascospores of Sclerotinia scierotiorum from air samples collected using a Burkard Multi-Vial Cyclone Sampler. The qPCR assay was highly sensitive and detected DNA from 0.5 to 5 x lO'' ascospores within a linear range (R"^ = 0.99). The qPCR assay was used to quantify ascospores of S. scierotiorum in air samples collected over three growing seasons. Initial SRC disease was observed 8 and 34 days following detection of 9.5 and 2 ascospores m ^ of air, respectively. Results from air samples collected using an Andersen N6 Sampler and the qPCR assay were compared with the BPT. Ascospore counts from a Burkard Sampler coupled with the qPCR assay and the BPT followed similar trends. In general, fewer ascospores were detected and bioaerosol sampling efficiency was low using an Anderson Sampler Three days were required to confirm the number of ascospores using SSM in the BPT and with an Andersen Sampler, whereas results from a Burkard Sampler coupled with the qPCR assay can provide results within 5 h of air sampling. The choice of method will depend on the available resources.

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“…The sensitivities of the assays were comparable to other real‐time PCR assays targeting the ITS or IGS regions of fungi or oomycetes (Chilvers et al . ; Blanco‐Meneses and Ristaino ; Catal et al . ; Chen et al .…”

Section: Discussionmentioning

confidence: 99%

Multiplex real-time PCR assays for detection of four seedborne spinach pathogens

et al. 2014

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Aims: To develop multiplex TaqMan real-time PCR assays for detection of spinach seedborne pathogens that cause economically important diseases on spinach. Methods and Results: Primers and probes were designed from conserved sequences of the internal transcribed spacer (for Peronospora farinosa f. sp. spinaciae and Stemphylium botryosum), the intergenic spacer (for Verticillium dahliae) and the elongation factor 1 alpha (for Cladosporium variabile) regions of DNA. The TaqMan assays were tested on DNA extracted from numerous isolates of the four target pathogens, as well as a wide range of nontarget, related fungi or oomycetes and numerous saprophytes commonly found on spinach seed. Multiplex real-time PCR assays were evaluated by detecting two or three target pathogens simultaneously. Singular and multiplex real-time PCR assays were also applied to DNA extracted from bulked seed and single spinach seed. Conclusions: The real-time PCR assays were species-specific and sensitive. Singular or multiplex real-time PCR assays could detect target pathogens from both bulked seed samples as well as single spinach seed. Significance and Impact of the Study: The freeze-blotter assay that is currently routinely used in the spinach seed industry to detect and quantify three fungal seedborne pathogens of spinach (C. variabile, S. botryosum and V. dahliae) is quite laborious and takes several weeks to process. The real-time PCR assays developed in this study are more sensitive and can be completed in a single day. As the assays can be applied easily for routine seed inspections, these tools could be very useful to the spinach seed industry.

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Detection and Quantification of Peronospora tabacina Using a Real-Time Polymerase Chain Reaction Assay

Cited by 19 publications

References 33 publications

Use of QuantitativePCRDetection Methods to Study Biocontrol Agents and Phytopathogenic Fungi and Oomycetes in Environmental Samples

Use of QuantitativePCRDetection Methods to Study Biocontrol Agents and Phytopathogenic Fungi and Oomycetes in Environmental Samples

Evaluation of Air Sampling and Detection Methods to Quantify Airborne Ascospores of Sclerotinia sclerotiorum

Multiplex real-time PCR assays for detection of four seedborne spinach pathogens

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