Subcellular localisation of cherry leaf roll virus coat genomic RNAs in tobacco leaves

The in vivo subcellular location of the coat protein and RNAs of cherry lea f roll nepovirus (CLRV) was studied in infected tobacco plants by two diffe rent approaches and it was correlated with the cytopathic structures induce d by the virus. Subcellular fractions were obtained by differential centrif ugation: visualised by electron microscopy and analysed for their viral RNA and coat protein content by Northern blot and Western blot analysis, respe ctively. Results indicate that viral RNAs accumulated preferentially at the microsomal fraction. Immunocytochemical studies revealed a clear associati on of the coat protein of CLRV with the virus-induced cytopathological stru ctures. In situ hybridisation studies confirmed the cytoplasmic location of the virus and allowed one to elucidate the distribution of the CLRV genomi c RNAs in the different cell types of infected tissue. (C) 2000 Elsevier Sc ience Ireland Ltd. All rights reserved.