CEREAL CHLOROTIC MOTTLE VIRUS - PURIFICATION, SEROLOGY AND ELECTRON-MICROSCOPY IN PLANT AND INSECT TISSUES

Cereal chlorotic mottle virus (CCMV) was shown to accumulate in the perinuclear space and cytoplasmic vesicles of infected plant cells. The virus was present in both vascular and mesophyll tissues. In infected leafhoppers Nesoclutha pa zlida (Evans) the virus particles had a similar appearance and intracellular location but sometimes appeared to be in a granular matrix and were dispersed throughout some degenerated nucleii. The virions (63 by 230 nm), when viewed in cross-section, showed 17 densely staining points around the circumference and had three concentric ring structures but no stained axial core. Tangential sections confirm the hexamer structure shown in negative stain. CCMV was purified by extraction in tris-HCI buffer and concentrated on a discontinuous sucrose gradient before filtering through a celite pad. The filtrate was pelleted and then given two sucrose gradient separations before the final pelleting. Purified virus was used to prepare an antiserum which had a titre of 128 and no detectable host reaction. In gel-diffusion tests with the homologous antiserum, CCMV produced two major reaction lines, with an additional line present in some tests. [Other keywords; rhabdovirus]. © 1979 ASEG.