Abstract
A simple, inexpensive technique has been needed to detect and distinguish the potato cyst nematodes Globodera rostochiensis and G. pallida, major pests of potatoes. We thus developed a sensitive duplex polymerase chain reaction (PCR) method using primers for the mitochondrial DNA’s cytochrome b gene region and agarose gel electrophoresis to identify G. rostochiensis and G. pallida and an efficient method to extract nematode DNA by crushing cysts in the presence of soil impurities. These methods provided the sensitivity to detect one egg of the target species, even if the extracted DNA solution was contaminated with PCR inhibitors derived from impurities. In addition, when the two species are both present in a sample at greatly different numbers, this method aided in detecting and identifying them accurately. The method will greatly benefit inspection institutions and advance the design of strategies to control these nematodes.
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Acknowledgements
This work was partially supported by grants from the Stabilization Fund Association of Hokkaido Potato Production and the Research Program on Development of Innovative Technology Grants from the Project of the Bio-oriented Technology Research Advancement Institution (BRAIN).
Funding
This work was funded by the Stabilization Fund Association of Hokkaido Potato Production and the Research Program on Development of Innovative Technology Grants from the Project of the Bio-oriented Technology Research Advancement Institution.
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Kushida, A., Sakai, H. Development of a simple method for simultaneous detection and differentiation of Globodera pallida and G. rostochiensis. J Gen Plant Pathol 88, 251–258 (2022). https://doi.org/10.1007/s10327-022-01065-6
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DOI: https://doi.org/10.1007/s10327-022-01065-6