Abstract
Rhizoctonia cerealis E.P. van der Hoeven (anamorph of Ceratobasidium cereale D.I. Murray and Burpee), which causes sharp eyespot in wheat, is a major soilborne fungal pathogen that severely impairs yield and quality of winter wheat in China. Because the pathogen is difficult to identify and quantify in soil using conventional methods, a rapid and reliable method is needed to detect and quantify the fungus. In this study, we developed an SYBR Green-based quantitative real-time polymerase chain reaction assay for specific, sensitive detection and quantification of R. cerealis in soil samples. Using a specific primer pair based on the β-tubulin gene of the fungal DNA sequence, we could specifically detect R. cerealis at quantities as low as 100 fg of purified pathogen DNA. Using the real-time PCR assays, we were able to quantify R. cerealis in artificially and naturally infested soil samples. This new technique to quantify R. cerealis is rapid and accurate and will be a useful tool for future studies of pathogenic R. cerealis.
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Acknowledgments
This research was supported by a fund earmarked for the China Agricultural Research System (CARS-3-1-17) and National Science Foundation of China (Grant 30900928). We also thank Professor Yu Jinfeng (College of Plant Protection, Shandong Agricultural University) for providing some standard isolates of binucleate Rhizoctonia.
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The nucleotide sequence data reported are available in the GenBank databases under accession numbers JF819943, JF819950 and JF819952.
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Guo, Y., Li, W., Sun, H. et al. Detection and quantification of Rhizoctonia cerealis in soil using real-time PCR. J Gen Plant Pathol 78, 247–254 (2012). https://doi.org/10.1007/s10327-012-0390-x
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DOI: https://doi.org/10.1007/s10327-012-0390-x