Validation of a real-time PCR for the quantitative estimation of a G143A mutation in the cytochrome bc(1), gene of Pyrenophora teres

Kianianmomeni A, Schwarz G, Felsenstein FG, Wenzel G (2007)
PEST MANAGEMENT SCIENCE 63(3): 219-224.

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Zeitschriftenaufsatz | Veröffentlicht | Englisch
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Abstract / Bemerkung
A single nucleotide polymorphism (SNP) in the cytochrome b gene confers resistance to strobilurin fungicides for several fungal pathogens. Therefore, on the basis of a change at amino acid position 143 from glycine to alanine, a real-time PCR assay was established for the quantitative detection of the analogous SNP in the cytochrome b sequence of Pyrenophora teres Drechsler, which causes barley net blotch. Allelic discrimination was achieved by using allele specific primers with artificially mismatched nucleic acid bases and minor groove binding probes. Validation parameters for the lower limits of the working range, namely limits of detection (LOD) and limits of quantification (LOQ), were statistically determined by the variance of calibration data, as well as by the variance of the 100% non-strobilurin-resistant allele DNA sample (blank values). It was found that the detection was limited by the variance of blank values (five in 801458 copies; 0.0006%), whereas the quantification was limited by the variance of calibration data (37 in 801458 copies; 0.0046%). The real-time PCR assay was finally used to monitor strobilurin-resistant cytochrome b alleles in barley net blotch field samples, which were already classified in in vivo biotests to be fully sensitive to strobilurins. All signals for strobilurin-resistant cytochrome b alleles were below the LOD, and therefore the results are in total agreement with the phenotypes revealed by biotests. (c) 2006 Society of Chemical Industry
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Zeitschriftentitel
PEST MANAGEMENT SCIENCE
Band
63
Zeitschriftennummer
3
Seite
219-224
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Kianianmomeni A, Schwarz G, Felsenstein FG, Wenzel G. Validation of a real-time PCR for the quantitative estimation of a G143A mutation in the cytochrome bc(1), gene of Pyrenophora teres. PEST MANAGEMENT SCIENCE. 2007;63(3):219-224.
Kianianmomeni, A., Schwarz, G., Felsenstein, F. G., & Wenzel, G. (2007). Validation of a real-time PCR for the quantitative estimation of a G143A mutation in the cytochrome bc(1), gene of Pyrenophora teres. PEST MANAGEMENT SCIENCE, 63(3), 219-224. doi:10.1002/ps.1290
Kianianmomeni, A., Schwarz, G., Felsenstein, F. G., and Wenzel, G. (2007). Validation of a real-time PCR for the quantitative estimation of a G143A mutation in the cytochrome bc(1), gene of Pyrenophora teres. PEST MANAGEMENT SCIENCE 63, 219-224.
Kianianmomeni, A., et al., 2007. Validation of a real-time PCR for the quantitative estimation of a G143A mutation in the cytochrome bc(1), gene of Pyrenophora teres. PEST MANAGEMENT SCIENCE, 63(3), p 219-224.
A. Kianianmomeni, et al., “Validation of a real-time PCR for the quantitative estimation of a G143A mutation in the cytochrome bc(1), gene of Pyrenophora teres”, PEST MANAGEMENT SCIENCE, vol. 63, 2007, pp. 219-224.
Kianianmomeni, A., Schwarz, G., Felsenstein, F.G., Wenzel, G.: Validation of a real-time PCR for the quantitative estimation of a G143A mutation in the cytochrome bc(1), gene of Pyrenophora teres. PEST MANAGEMENT SCIENCE. 63, 219-224 (2007).
Kianianmomeni, Arash, Schwarz, Gerhard, Felsenstein, Friedrich G., and Wenzel, Gerhard. “Validation of a real-time PCR for the quantitative estimation of a G143A mutation in the cytochrome bc(1), gene of Pyrenophora teres”. PEST MANAGEMENT SCIENCE 63.3 (2007): 219-224.

9 Zitationen in Europe PMC

Daten bereitgestellt von Europe PubMed Central.

Allele-specific PCR for quantitative analysis of mutants in live viral vaccines.
Bidzhieva B, Laassri M, Chumakov K., J Virol Methods 201(), 2014
PMID: 24607431
Assessment of fungicide resistance and pathogen diversity in Erysiphe necator using quantitative real-time PCR assays.
Dufour MC, Fontaine S, Montarry J, Corio-Costet MF., Pest Manag Sci 67(1), 2011
PMID: 20949585
Real-time PCR Assay with SNP-specific Primers for the Detection of a G143A Mutation Level in Venturia inaequalis Field Populations
Michalecka M, Malinowski T, Broniarek-Niemiec A, Bielenin A., Journal of phytopathology. 159(7-8), 2011
PMID: IND44598503
Evaluation of single-base substitution rate in DNA by affinity capillary electrophoresis.
Kanayama N, Takarada T, Shibata H, Kimura A, Maeda M., Anal Chim Acta 619(1), 2008
PMID: 18539181

26 References

Daten bereitgestellt von Europe PubMed Central.

Characterizing resistance risk of Erysiphe graminis f.sp. tritici to strobilurins.
Chin KM, Chavaillaz D, Kaesbohrer M, Staub T, Felsenstein FG., Crop Prot. 20(2), 2001
PMID: IND22435391

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