Quantification of Tilletia caries and Tilletia controversa mycelium in wheat apical meristem by real-time PCR.
TL;DR: 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.
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Abstract: Zouhar M., Mazakova J., Prokinova E., Vaňova M., Rysanek P. ( 2010): Quantification of Tilletia caries and Tilletia controversa mycelium in wheat apical meristem by real-time PCR. Plant Protect. Sci., 46: 107-115. In the Czech Republic, three closely related species of the genus Tilletia belong to pathogens that cause signifi - cant 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 r 2 = 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.
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Citations
Assessment of the loose smut fungi (Ustilago nuda and U. tritici) in tissues of barley and wheat by fluorescence microscopy and real-time PCR
TL;DR: Results indicated that a prediction of loose smut infection by real-time PCR is possible at the second leaf stage, and that the assay is equally suited for use with spring and winter varieties of barley and wheat.
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Detection of Tilletia caries, Tilletia laevis and Tilletia controversa wheat grain contamination using loop-mediated isothermal DNA amplification (LAMP).
TL;DR: The presented data showed that the LAMP analysis is a simple, specific and rapid method that could be used for detection of Tilletia spp.
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Development of the Droplet Digital PCR to Detect the Teliospores of Tilletia controversa Kühn in the Soil With Greatly Enhanced Sensitivity.
TL;DR: This study was the first report using ddPCR techniques to detect T. controversa teliospores in soil with greatly enhanced sensitivity and this sensitivity was 100 times more sensitive than that of simple PCR.
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Methyljasmonate and salicylic acid contribute to the control of Tilletia controversa Kühn, causal agent of wheat dwarf bunt
TL;DR: It is suggested that Meja and SA display a distinct role in activation of defence genes in the roots and coleoptiles and that they eliminate the fungal pathogen movement to upper parts of the plants with the passage of time as the anthers mature.
Development of a loop-mediated isothermal amplification assay for the detection of Tilletia controversa based on genome comparison.
Somayyeh Sedaghatjoo,Monika K. Forster,Ludwig Niessen,Petr Karlovsky,Berta Killermann,Wolfgang Maier +5 more
TL;DR: In this article, a loop-mediated isothermal amplification (LAMP) was developed based on one of these DNA regions, which was verified using 223 fungal samples comprising 43 fungal species including 11 Tilletia species.
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