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Plant Pathology Journal
Year: 2006  |  Volume: 5  |  Issue: 3  |  Page No.: 378 - 382

Specific Detection of Peronospora tabacina by PCR-amplified rDNA Sequences

Jwu-Guh Tsay, Chishih Chu, Ya-Han Chuang and Ruey-Shyang Chen    

Abstract: Peronospora tabacina Adam, a downy mildew fungus, is a devastating disease of tobacco and a pathogen of import prohibition in Taiwan. For quarantine purpose, we developed a nested PCR method to detect this pathogen. With universal primer pair ITS1/ITS4, internal transcribed spacer (ITS) region of pathogen`s rDNA was amplified. Specific PCR primers (PT1/PT2) were designed based on ITS sequence and used to amplify a 493-bp rDNA fragments from P. tabacina. In order to increase the sensitivity, another primer pair (PT3/PT4) was used in a one-tube nested PCR. The expected 243-bp fragment was amplified from P. tabacina, but not from any other tested fungi or commercial tobacco leaves. The detection limit of this nested PCR could be low as 1 pg of template DNA. The present nested PCR method showed to be rapid, simple and available as a tool to screen P. tabacina infection for quarantine purpose.

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