Quantification of Tilletia caries and Tilletia controversa mycelium in wheat apical meristem by real-time PCR

Miloslav Zouhar, Jana Mazáková, Evženie Prokinová, Marie Váňová and Pavel Ryšánek
Additional contact information
Miloslav Zouhar: Department of Plant Protection, Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences Prague, Prague-Suchdol, Czech Republic
Jana Mazáková: Department of Plant Protection, Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences Prague, Prague-Suchdol, Czech Republic
Evženie Prokinová: Department of Plant Protection, Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences Prague, Prague-Suchdol, Czech Republic
Marie Váňová: Agricultural Research Institute Kroměříž, Ltd., Kroměříž, Czech Republic
Pavel Ryšánek: Department of Plant Protection, Faculty of Agrobiology, Food and Natural Resources, Czech University of Life Sciences Prague, Prague-Suchdol, Czech Republic

Plant Protection Science, 2010, vol. 46, issue 3, 107-115

Abstract: In the Czech Republic, three closely related species of the genus Tilletia belong to pathogens that cause significant losses of wheat crops by replacing grains with a mass of teliospores. A quantitative real-time qPCR assay using SYBR Green I has been developed to quantify the amount of T. caries and T. controversa mycelium in apical meristems of different wheat varieties. The real-time PCR reaction was validated by evaluating selected extraction methods, examining the specificity of designed target-specific IGS primers and verifying the optimised amplification reaction on naturally infected wheat plants. The PCR detection limit for the specific identification of fungal DNA was 0.22 ng of mycelium, and the negative correlation between the target DNA quantity and cycle threshold (Ct) was high with a coefficient of determination of R2 = 0.992. The developed method was used to quantify pathogens mycelium in five wheat varieties in the range from 0.34 ng to 15 µg per one growing tip.

Keywords: Tilletia spp.; real-time PCR; apical meristem; absolute quantification (search for similar items in EconPapers)
Date: 2010
References: View complete reference list from CitEc
Citations: View citations in EconPapers (1)

Downloads: (external link)
http://pps.agriculturejournals.cz/doi/10.17221/50/2009-PPS.html (text/html)
http://pps.agriculturejournals.cz/doi/10.17221/50/2009-PPS.pdf (application/pdf)
free of charge

Related works:
This item may be available elsewhere in EconPapers: Search for items with the same title.

Export reference: BibTeX RIS (EndNote, ProCite, RefMan) HTML/Text

Persistent link: https://EconPapers.repec.org/RePEc:caa:jnlpps:v:46:y:2010:i:3:id:50-2009-pps

DOI: 10.17221/50/2009-PPS

Access Statistics for this article

Plant Protection Science is currently edited by Ing. Eva Karská, (Executive Editor PPS)

More articles in Plant Protection Science from Czech Academy of Agricultural Sciences
Bibliographic data for series maintained by Ivo Andrle ().