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Development of a multiplex PCR method for simultaneous detection of diagnostic DNA markers of five disease and pest resistance genes in potato

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Abstract

Multiplex PCR is practically a reasonable choice for molecular marker-assisted selection in potato breeding. We had developed and were using a multiplex PCR method for selection of resistance genes to cyst nematode (H1), Potato virus X (Rx1) and late blight (R1 and R2). Since then, more reliable and tightly linked markers for H1 and R2, and a new marker for resistance to Potato virus Y (Ry chc ) were developed. In this article, all these superior markers, including a positive marker to eliminate PCR-failed samples, were incorporated into one multiplex PCR assay. Using the newly developed multiplex PCR technique, five plants potentially harboring all five resistance genes were selected from 96 hybrid plants approximately 5 h after DNA extraction, which is a third of the operation time compared with separate PCR reactions for each marker.

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Acknowledgments

This study was supported in part by the Ministry of Agriculture, Forestry and Fisheries of Japan (Rural Bio-Mass Research Project, BCD-11232). We highly appreciate Tetsuo Maoka and Takashi Narabu, National Agricultural Research Center for Hokkaido Region, Tetsuji Ogawa, Nagasaki Agricultural and Forestry Technical Development Center, and Mikiko Kawahara and Yasuhiro Ogawa, Nagasaki Pest Control Station, for performance of or assistance in biological assays of disease resistances.

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Authors and Affiliations

  1. Nagasaki Agricultural and Forestry Technical Development Center, Unzen, Nagasaki, 854-0302, Japan

    Kazuyuki Mori, Yu Sakamoto, Nobuhiro Mukojima & Takashi Nakao

  2. Graduate School of Agricultural Science, Kobe University, Nada, Kobe, Hyogo, 657-8501, Japan

    Kazuyuki Mori & Takashige Ishii

  3. National Agricultural Research Center for Hokkaido Region, Memuro, Kasai, Hokkaido, 082-0081, Japan

    Seiji Tamiya & Kazuyoshi Hosaka

Authors
  1. Kazuyuki Mori
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  2. Yu Sakamoto
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  3. Nobuhiro Mukojima
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  4. Seiji Tamiya
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  5. Takashi Nakao
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  6. Takashige Ishii
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  7. Kazuyoshi Hosaka
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Correspondence to Kazuyoshi Hosaka.

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Mori, K., Sakamoto, Y., Mukojima, N. et al. Development of a multiplex PCR method for simultaneous detection of diagnostic DNA markers of five disease and pest resistance genes in potato. Euphytica 180, 347–355 (2011). https://doi.org/10.1007/s10681-011-0381-6

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  • DOI: https://doi.org/10.1007/s10681-011-0381-6

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