Biotechnology & Biotechnological Equipment

Volume 21, Number 1
Date: 2007

DIRECT PCR FOR DNA BARCODING IN THE GENERA PHYTOPHOPTHORA AND PYTHIUM

G. Calmin, L. Belbahri and F. Lefort

Abstract

A protocol for direct polymerase chain reaction (DPCR) amplification of commonly used phylogenetic markers of the genera Pythium and Phytophthora from mycelia was developed. This has proven useful for direct PCR amplification from hypha, zoospores and infected plant material, which is quicker and simpler, than other proposed attempts. Optimal reaction conditions are described allowing amplification of commonly used phylogenetic markers with a size of up to 1 kbp. This approach proved successful for amplification of selected markers from hundreds of Phytophthora, Pythium and fungal isolates and has been so far used in routine genotyping identification of oomycota and fungi in our laboratory.

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