VIEW ARTICLE

Techniques

A Purification Procedure for Enhancement of Citrus Tristeza Virus Yields and its Application to Other Phloem-Limited Viruses. R. F. Lee, Associate professor, University of Florida, Citrus Research and Education Center, Lake Alfred 33850; S. M. Garnsey(2), R. H. Brlansky(3), and A. C. Goheen(4). (2)Research plant pathologist, U.S. Department of Agriculture, Agricultural Research Service, Orlando, FL 32803; (3)Associate professor, University of Florida, Citrus Research and Education Center, Lake Alfred 33850; (4)U.S. Department of Agriculture, University of California, Davis 95616. Phytopathology 77:543-549. Accepted for publication 18 July 1986. Copyright 1987 The American Phytopathological Society. DOI: 10.1094/Phyto-77-543.

An improved purification procedure for increased yield and recovery of intact particles of citrus tristeza virus (CTV) involves the use of polyethylene glycol p-isooctylphenyl ether in the extraction buffer, two polyethylene glycol precipitation steps, and centrifugation on a preformed step isopycnic cesium sulfate gradient. The quantity and quality of CTV was monitored throughout the purification procedure by enzyme-linked immunosorbent assay (ELISA) and serologically specific electron microscopy. The ELISA with antisera prepared against unfixed CTV was better at detection of short length CTV particles from cesium sulfate gradient zones than ELISA with antisera prepared against formaldehyde-fixed CTV. Use of this purification procedure yielded as high as 31% intact CTV particles in purified infectious preparations. The procedure was also used to assay for virus-like particles from Nandina domestica ‘Nanapurpurea’ and from grapevines affected with corky bark disease, both of which are suspected to be infected by phloem-limited closteroviruses.