Japanese Journal of Phytopathology
Online ISSN : 1882-0484
Print ISSN : 0031-9473
ISSN-L : 0031-9473
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Volume 79, Issue 2
Displaying 1-4 of 4 articles from this issue
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Original
  • Detection and absolute quantification of Xanthomonas axonopodis pv. glycines from soybeans by real-time PCR.
    T. WATANABE, H. SAWADA
    2013 Volume 79 Issue 2 Pages 83-91
    Published: 2013
    Released on J-STAGE: June 01, 2013
    DOIhttps://doi.org/10.3186/jjphytopath.79.83
    JOURNAL FREE ACCESS
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    A real-time PCR assay (qPCR) based on TaqMan chemistry was developed to quantify Xanthomonas axonopodis pv. glycines, the causal agent of bacterial pustule of soybeans. Specific primers and probes were designed based on the sequences of the rpoD gene. In the assay, the gene from 61 X. axonopodis pv. glycines strains was amplified, but not from any other Xanthomonas species tested, various reference strains derived from soybeans, or 30 indigenous bacteria isolated from soybeans in Fukui Prefecture. For absolute quantification, a calibration curve was constructed based on a recovery test (Sawada et al., Plant Protect. 62: 611-617, 2008): a series of 1 g soybean seeds was spiked with 10-fold dilutions of the bacterial suspensions, and DNA from each 1-g series with a 10-fold dilution was extracted and subjected to qPCR. A strong linear relationship was found between the threshold cycle value and the bacterial density from 6×102 to 6×108 cfu per 1 g of soybean seeds; the detection limit might be close to 6×101 cfu per 1 g of seeds. This qPCR experimental system in conjunction with the following sampling procedure proved to be a reliable method for estimating the degree of contamination in seeds: (1) crush 1 kg of test seeds and mix well, (2) sample 1 g of these crushed seeds, and repeat this procedure 10 times, (3) extract DNA from every 1 g of crushed seeds sampled and perform the qPCR experiment three times to derive quantitative values, (4) average the 30 quantitative values obtained, and use this value to determine the degree of contamination of the original seeds tested.
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Short Communications
  • Combined treatment with chemicals and dry heat to disinfest cucurbit seeds after infestation with Acidovorax citrulli.
    M. KUBOTA, K. HARA, N. HAGIWARA, T. SHIRAKAWA
    2013 Volume 79 Issue 2 Pages 92-98
    Published: 2013
    Released on J-STAGE: June 01, 2013
    DOIhttps://doi.org/10.3186/jjphytopath.79.92
    JOURNAL FREE ACCESS
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    A mixture of CuSO4 and acetic acid disinfested wax gourd seeds that had been infested with Acidovorax citrulli more effectively than did ascorbic acid, peroxyacetic acid or a mixture of oxytetracycline, streptomycin and CuSO4. Seeds of melon, cucumber and wax gourd were completely disinfested after a 60-min soak in the mixture followed by dry-heat at 75°C for 4 days without affecting germination. Seeds of brown seed squash, large seed squash and bottle gourd were disinfested by a 60-min soak in the mixture followed by dry-heat at 80°C for 4 days, which caused a small delay in germination. Any treatments of small or white seeds of squash that combined a soak in the chemical mixture with dry heat and effectively disinfested the seeds also inhibited germination.
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  • Fruit rot, a new symptom of okra bacterial leaf blight, caused by Pseudomonas cichorii.
    T. INOUE, H. KAJIHARA, K. MURAMOTO, R. YOSHIOKA, H. SAWADA
    2013 Volume 79 Issue 2 Pages 99-104
    Published: 2013
    Released on J-STAGE: June 01, 2013
    DOIhttps://doi.org/10.3186/jjphytopath.79.99
    JOURNAL FREE ACCESS
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    Fruit rot of a local variety of okra [Abelmoschus esculentus (L.) Moench], Shiro Okura, has been observed in Yamaguchi Prefecture, Japan since 2008. The causal bacterium was identified as Pseudomonas cichorii, a pathogen causing okra bacterial leaf blight, according to its phenotypic characteristics and 16S rDNA-based phylogenetic analysis. When Shiro Okura fruits were inoculated with the isolates, the fruit rot was reproduced, and the bacterium was reisolated from the plants. To our knowledge, this is the first report of fruit rot of okra caused by P. cichorii. We propose adding fruit rot as a new symptom of okra bacterial leaf blight.
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