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Quantification of Rice Blast Disease Progressions Through Taqman Real-Time PCR

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Abstract

Rice blast caused by Magnaporthe oryzae is a major disease in the paddy field and also a representative model system in the investigation of plant–microbe interactions. This study was undertaken to provide the quantitative evaluation method that specifically determines the amount of M. oryzae proliferation in planta. Real-time PCR was used as the detection strategy in combination with the primer pair and Taqman probe specific to MHP1, a unigene encoding HYDROPHOBIN that is indispensable for normal virulence expression. Based on the crossing point values from the PCR reactions containing a series of increasing concentration of cloned amplicon or fungal genomic DNA, correlation among the template’s copy number or its amount and amplification pattern was calculated. Reliability of this equation was further confirmed using the DNA samples from the rice leaves infected with compatible or incompatible strains of M. oryzae. The primer pair used in the Taqman real-time PCR reaction can recognize the existence of fungal DNA as low as 1 pg. In sum, our quantitative evaluation system is applicable and reliable in the blast diagnosis and also in the estimation of objective blast disease progression.

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Acknowledgments

This research was supported by grants from the Research Program for Agricultural Science and Technology Development (Project No. PJ008840) and the Next-Generation BioGreen 21 Program (Plant Molecular Breeding Center No. PJ008021072011), Rural Development Administration, Korea.

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Authors and Affiliations

  1. Rural Development Administration, National Academy of Agricultural Science, Suwon, 441-707, South Korea

    Mukhamad Su’udi, Jinyeong Kim, Jong-Mi Park, Shin-Chul Bae, Donghern Kim, Yong-Hwan Kim & Il-Pyung Ahn

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  1. Mukhamad Su’udi
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  2. Jinyeong Kim
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Correspondence to Il-Pyung Ahn.

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Su’udi, M., Kim, J., Park, JM. et al. Quantification of Rice Blast Disease Progressions Through Taqman Real-Time PCR. Mol Biotechnol 55, 43–48 (2013). https://doi.org/10.1007/s12033-012-9632-6

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  • DOI: https://doi.org/10.1007/s12033-012-9632-6

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