In situ detection of early replication phases of a gemini virus in legume protoplasts |
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| Authors: | Dale M. Steffensen H. J. Wilson Robert M. Goodman |
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| Affiliation: | (1) Department of Genetics and Development, University of Illinois, 61801 Urbana, IL, USA;(2) Department of Biological Sciences, University of Alabama at Huntsville, 35899 Huntsville, AL, USA;(3) Calgene, Incorporated, 95616 Davis, CA, USA |
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| Abstract: | Protoplasts of Phaseolus vulgaris L. (Top Crop), infected with bean golden mosaic virus, were isolated and fixed by various methods for in situ hybridization. An iodine-125 labeled probe was made from the replicative form of the virus. The localization and quantitation was done by autoradiography. Cell wall removal lowered the background and allowed a more accurate analysis. RNase was used to eliminate the possibility of hybrids to RNA. The evidence suggests a sequence of virus movements starting from rough endoplasm reticulum, moving to the nuclear membrane, and finally with the highest concentration inside the nucleus.Abbreviations BGMV bean golden mosaic virus - rfBGMV or rfDNA replicative double-stranded DNA virus - ssDNA single-stranded virus |
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