Journal of Microbiology

The Microbiological Society of Korea (한국미생물학회)
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Selection of RAPD Markers for Phytophthora infestans and PCR Detection of Phytophthora infestans from Potatoes

  • Kim, Kyung-Su (Plant Microbiology and MPMI Laboratory, Division of Applied Plant Sciences, College A Agriculture and Life Science, Kangwon National University) ;
  • Lee, Youn-Su (Plant Microbiology and MPMI Laboratory, Division of Applied Plant Sciences, College A Agriculture and Life Science, Kangwon National University)
  • Published : 2001.06.01
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Abstract

For rapid and secure differentiation of P. infestans from other Phytophthora species, two fragments obtained from randomly amplified polymorphic DNA (RAPD) profiles were selected as markers. Also, primers for in polymerase chain reaction (PCR) to detect P. infestans specifically were developed by analyzing the sequences of ITSII regions in rDNA of Phytophthora species. The primers, PISP-1 and ITS3 amplified a single. Fragment 450 bp of about in P. infestans, but not in other fungal or bacterial isolates. Annealing temperatures and template DNA quantities were varied for the optimization of PCR conditions. From the result of the PCR detection study, species-specific primers were selected under annealing temperatures ranging from 55$^{\circ}C$ to 61$^{\circ}C$, and template DNA levels ranging from 10 pg to 100 ng.

Keywords

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