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Development of Multiplex PCR to Detect Five Pythium Species Related to Turfgrass Diseases

Journal of Phytopathology (Impact Factor: 1). 08/2010; 158(9):609 - 615. DOI: 10.1111/j.1439-0434.2009.01660.x

ABSTRACT The objective of this study was to develop multiplex PCR detection method for five Pythium species associated with turfgrass diseases, Pythium aphanidermatum, Pythium arrhenomanes, Pythium graminicola, Pythium torulosum and Pythium vanterpoolii. Species-specific primers and two common primers were designed based on the sequences of the internal transcribed spacer region of ribosomal DNA. Another primer set by which all organisms would be amplified in 18S rDNA was used as a positive control. When these total nine primers were applied to the multiplex PCR, all species were individually discriminated in the mixture of five species culture DNA. Furthermore, all five Pythium species were detected in naturally infected plants using the multiplex PCR.

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