A technique for the extraction and purification of viable Plasmodiophora brassicae resting spores from host root tissue

Abstract

An improved technique for purifying Plasmodiophora brassicae resting spores from host root tissue and other contaminants is described. After extraction from macerated infected Chinese cabbage roots by a series of centrifugations in 50% sucrose and distilled water, P. brassicae resting spores were separated from other contaminants in a continuous gradient of LUDOX (40% silica suspended in NaOH, pH 9.8; DuPont)/deionized water. This purification technique produces clean suspensions of P. brassicae resting spores suitable for propagation of the organism, long-term storage, and extraction of DNA or other molecules of interest.

Key Words:

• clubroot
• Plasmodiophora brassicae
• Plasmodiophoromycetes
• purification