Comparison of renografin density gradient centrifugation and ion-exchange chromatography for purification of phony peach bacterium from plant extracts

Abstract

Renografin density gradient centrifugation and carboxymethyl (CM)-and diethylaminoethyl (DEAE)-cellulose ion-exchange chromatography were compared for purification of cells of phony peach, rickettsia-like bacterium (RLB) from plant extracts. Centrifugation on 30–40% linear gradients resulted in four bands. Cells of RLB sedimented into the third band located 4.9–5.4 cm below the meniscus. Phase-contrast microscopy, plant peroxidase activity, and viable bacterial assays showed the resulting cells ofRLB to be relatively free of host contaminants. A typical purification from 120 ml of extract containing 7.2×109 RLB cells resulted in the recovery of 7.0×107 RLB cells. Cells ofRLB were not successfully purified byCM andDEAE chromatography. Unitil a medium is available for cultivation of plantRLB, Renografin density gradient centrifugation should be a useful tool for obtaining cells ofRLB for biochemical and serological investigations.