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Simple and sensitive BIO-PCR detection of potato blackleg pathogens from stem, tuber, and soil samples

  • Bacterial and Phytoplasma Diseases
  • Published:
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Abstract

A BIO-PCR was developed for the detection of potato blackleg pathogens, including Pectobacterium wasabiae (Pw), P. atrosepticum (Pa), P. carotovorum subsp. brasiliense (Pcb), and Dickeya dianthicola (Ddi). The method was based on the semi-selective incubation of pathogens using pectate-based liquid enrichment medium (LEM) and PCR detection of blackleg pathogens (triplex PCR for Pw, Pa, and Ddi and simplex PCR for Pcb with previously reported primers). Incubation in LEM for 24–120 h at 25 °C was effective in detecting the pathogens from plants (stems and tubers) and the soil around the roots. The detection limit by BIO-PCR from the tuber sample (0.6 g) in 10 ml medium was 2–8 colony-forming units (cfu) for four pathogens, whereas that from soil (10 g) in 100 ml medium was 2 × 102, 3 × 103, 6 × 101, and 2 × 101 cfu for Pw, Pa, Pcb, and Ddi, respectively. All four blackleg pathogens could also be detected from symptomatic stems, progeny tubers, and the soil around the stem base of diseased plants by BIO-PCR. This method is a promising technique for the efficient detection of potato blackleg pathogens not only for seed inspection but also for epidemiological research.

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Acknowledgements

This work was supported by the Science and Technology Research Promotion Program for the Agriculture, Forestry, Fisheries, and Food industry (no. 27005C). We thank Nobuya Onozuka (Hokkaido Agricultural Research Center, NARO) for valuable comments on the manuscript.

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Authors and Affiliations

  1. Hokkaido-Chuo Station, Center for Seeds and Seedlings (NCSS), National Agriculture and Food Research Organization (NARO), Nishinosato, Kitahiroshima, Hokkaido, 061-1102, Japan

    Yoshiyuki Aono

  2. Graduate School of Agriculture, Hokkaido University, N9 W9 Kita-ku, Sapporo, Hokkaido, 060-8589, Japan

    Yoshiyuki Aono, Takehiro Ohki & Norikuni Oka

  3. Hokkaido Agricultural Research Center, NARO, Hitsujigaoka, Toyohira-ku, Sapporo, Hokkaido, 062-8555, Japan

    Takato Nakayama, Taketo Fujimoto, Takehiro Ohki, Norikuni Oka & Tetsuo Maoka

  4. Tokachi Federation of Agricultural Cooperatives, W3 S7, Obihiro, Hokkaido, 080-0013, Japan

    Takahiro Ozawa

  5. Tokachi Station, NCSS, NARO, Koufuku, Obihiro, Hokkaido, 089-1246, Japan

    Yutaka Ushio

  6. Donan Agricuitural Experiment Station, Hokkaido Research Organization, Hon-cho, Hokuto, Hokkaido, 041-1201, Japan

    Shinji Yasuoka

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  1. Yoshiyuki Aono
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  2. Takato Nakayama
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  3. Takahiro Ozawa
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  4. Yutaka Ushio
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  5. Shinji Yasuoka
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  6. Taketo Fujimoto
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  7. Takehiro Ohki
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  9. Tetsuo Maoka
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Correspondence to Yoshiyuki Aono.

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Aono, Y., Nakayama, T., Ozawa, T. et al. Simple and sensitive BIO-PCR detection of potato blackleg pathogens from stem, tuber, and soil samples. J Gen Plant Pathol 87, 209–218 (2021). https://doi.org/10.1007/s10327-021-00997-9

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  • DOI: https://doi.org/10.1007/s10327-021-00997-9

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