VIEW ARTICLE

Techniques

Purification of Citrus Tristeza Virus from Diseased Citrus Fruits and the Detection of the Virus in Citrus Tissues by Fluorescent Antibody Techniques. T. Tsuchizaki, Plant Pathologist, Institute for Plant Virus Research, Tukuba Science City, Yatabe, Ibaraki, 300-21, Japan; A. Sasaki(2), and Y. Saito(3). (2)Plant Pathologist, Hiroshima Prefecture Fruit Tree Experimental Station, Kiharacho, Mihara, Hiroshima, 729-03, Japan; (3)Plant Pathologist, Institute for Plant Virus Research, Tukuba Science City, Yatabe, Ibaraki, 300-21, Japan. Phytopathology 68:139-142. Accepted for publication 21 July 1977. Copyright © 1978 The American Phytopathological Society, 3340 Pilot Knob Road, St. Paul, MN 55121. All rights reserved.. DOI: 10.1094/Phyto-68-139.

Citrus tristeza virus (CTV) can be purified more easily from diseased fruits than from bark. The purification procedure was: gentle grinding of the pericarp without flavedo of Hassaku or navel orange fruits, clarification with carbon tetrachloride, polyethylene glycol precipitations, and a sucrose density gradient centrifugation. It was shown by electron microscopy that 50% of the particles in the purified preparations were of the normal length (2,000 nm). The ratio of UV absorption at 260 nm and 280 nm (A_260/280) for purified virus was 1.22. Antiserum against the Hassaku dwarf strain of CTV was prepared in a rabbit given one intravenous and three intramuscular injections of purified preparations. The antiserum reacted to the same dilution end points in complement fixation tests with three strains of CTV: the Hassaku dwarf strain, the seedling yellows strain, and the mild strain. Fluorescent antibodies of the Hassaku dwarf strain of CTV were used for the detection of CTV within tissues of several citrus species. This technique may become a useful tool for the detection of CTV in citrus tissues.