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Journal of Virology, November 2002, p. 10654-10664, Vol. 76, No. 21
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.21.10654-10664.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

In Situ Localization and Tissue Distribution of the Replication-Associated Proteins of Cucumber Mosaic Virus in Tobacco and Cucumber

Fabrizio Cillo,{dagger} Ian M. Roberts, and Peter Palukaitis*

Scottish Crop Research Institute, Invergowrie, Dundee DD2 5DA, United Kingdom

Received 13 March 2002/ Accepted 22 July 2002

The replication-associated proteins encoded by Cucumber mosaic virus (CMV), the 1a and 2a proteins, were detected by immunogold labeling in two host species of this virus, tobacco (Nicotiana tabacum) and cucumber (Cucumis sativus). In both hosts, the 1a and 2a proteins colocalized predominantly to the vacuolar membranes, the tonoplast. While plus-strand CMV RNAs were found distributed throughout the cytoplasm by in situ hybridization, minus-strand CMV RNAs were barely detectable but were found associated with the tonoplast. In both cucumber and tobacco, 2a protein was detected at higher densities than 1a protein. The 1a and 2a proteins also showed quantitative differences with regard to tissue distributions in tobacco and cucumber. About three times as much 2a protein was detected in CMV-infected cucumber tissues as in CMV-infected tobacco tissues. In tobacco, high densities of these proteins were observed only in vascular bundle cells of minor veins. In contrast, in cucumber, high densities of 1a and 2a proteins were observed in mesophyll cells, followed by epidermis cells, with only low levels being observed in vascular bundle cells. Differences were also observed in the distributions of 2a protein and capsid protein in vascular bundle cells of the two host species. These observations may represent differences in the relative rates of tissue infection in different hosts or differences in the extent of virus replication in vascular tissues of different hosts.

* Corresponding author. Mailing address: Scottish Crop Research Institute, Invergowrie, Dundee DD2 5DA, United Kingdom. Phone: 44 1382 568523. Fax: 44 1382 568575. E-mail: ppaluk{at}scri.sari.ac.uk.

{dagger} Present address: Centro di Studio del C.N.R. sui Virus e le Virosi delle Colture Mediterranee, 70126 Bari, Italy.

Journal of Virology, November 2002, p. 10654-10664, Vol. 76, No. 21
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.21.10654-10664.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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