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A PCR-based assay for differentiating A- and A*-type strains of Xanthomonas citri subsp. citri, the causal agent of Asiatic citrus canker

  • Bacterial and Phytoplasma Diseases
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Abstract

By applying A- and A*-type strains of Xanthomonas citri subsp. citri (Xcc) in a repetitive sequence-based polymerase chain reaction (rep-PCR), two DNA amplicons, one unique to each strain, were evaluated as a probe against the DNA of Xcc strains. Two pairs of primers derived from these amplicons were tested in a PCR analysis. The results confirmed that primers Ms+/Ms− are useful for differentiating A-type from A*-type strains of Xcc. Also, a multiplex PCR with both set of primers can be used to distinguish three groups in Xcc populations: A-type strains and two subgroups of A* strains including Iranian and Thai A* strains.

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Acknowledgments

Most of this study was performed in the Citrus Research Division, Kuchinotsu, NARO Institute of Fruit Tree Science, Nagasaki, Japan.

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Authors and Affiliations

  1. Department of Plant Protection, College of Agriculture, Shiraz University, Shiraz, Iran

    Fatemeh Yousefi Koupaei & S. Mohsen Taghavi

  2. Citrus Research Division, Kuchinotsu, NARO Institute of Fruit Tree Science, 954 Otsu, Kuchinotsu, Minamishimabara, Nagasaki, 859-2501, Japan

    Hiroshi Shiotani

  3. Faculty of Agriculture, Shizuoka University, 836 Ohya, Suruga-ku, Shizuoka, 422-8529, Japan

    Shinji Tsuyumu

Authors
  1. Fatemeh Yousefi Koupaei
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  2. S. Mohsen Taghavi
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  3. Hiroshi Shiotani
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  4. Shinji Tsuyumu
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Corresponding author

Correspondence to Hiroshi Shiotani.

Electronic supplementary material

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Fig. 1 Specificity of Ms+/Ms− based PCR assay for the detection of A* type of Xanthomonas citri subsp. citri strains. Lanes 1–8: Xcc-A* strains (nos. 88, 16, 50, 85, 101, 102, 103, 110, respectively), Lanes 9–18: Xcc-A strains (KC1, KC5, KC7, KC10, KC19, KC20, KC21, KC31, KC38). Lane M: DNA ladder mix marker. IR-A*: Iranian A* strains, Thai-A*: A* strains from Thailand.

Fig. 2 PCR amplification with M+/M− primers of DNA from representative strains of Xanthomonas citri subsp. citri. Product of expected size has been amplified for A* strains from Thailand and A strains, but no amplification of Iranian A* strains. Lanes 1–7 and 12–15: Iranian strains nos. 1, 32, 34, 51, 67, 85, 95 and 103, 105, 107, 110, respectively. Lanes 8–11: A* strains from Thailand (nos. 97, 98, 100 and 101, respectively). Lanes 16–22: Xcc-A strains (KC3, KC5, KC20, KC21, KC26, KC38 and NCCPB409, respectively). Lane M: DNA ladder mix marker. IR-A*: Iranian A* strains, Thai-A*: A* strains from Thailand.

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Koupaei, F.Y., Taghavi, S.M., Shiotani, H. et al. A PCR-based assay for differentiating A- and A*-type strains of Xanthomonas citri subsp. citri, the causal agent of Asiatic citrus canker. J Gen Plant Pathol 80, 85–89 (2014). https://doi.org/10.1007/s10327-013-0494-y

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  • DOI: https://doi.org/10.1007/s10327-013-0494-y

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