Abstract
We developed real-time PCR assays using TaqMan probes to detect and quantify Rosellinia necatrix, the causal agent of white root rot in many plant species. Two sets of PCR primers and TaqMan probe indicated that their detection limits could be as low as 1 fg of template DNA. Using the real-time PCR assays with the TaqMan probes, we were able to quantify R. necatrix DNA in naturally diseased roots of Japanese pear and in artificially infested soil samples. Although the new assays were inadequate for use with naturally infested soil samples, nested PCR procedures improved the detectability of the new assays.
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Acknowledgments
We thank Drs. T. Usami and K. Ohkawa for valuable comments. This study was supported in part by a Grant-in-Aid for Scientific Research (21580052) from the Japan Society for the Promotion of Science.
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Shishido, M., Kubota, I. & Nakamura, H. Development of real-time PCR assay using TaqMan probe for detection and quantification of Rosellinia necatrix in plant and soil. J Gen Plant Pathol 78, 115–120 (2012). https://doi.org/10.1007/s10327-012-0366-x
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DOI: https://doi.org/10.1007/s10327-012-0366-x