Japanese Journal of Phytopathology
Online ISSN : 1882-0484
Print ISSN : 0031-9473
ISSN-L : 0031-9473
Detection of Cucumber Mosaic Virus RNA5 by Hybridization with Photobiotin-Labelled Double-stranded RNA5 Probe
Wei Qin WANGTomohide NATSUAKISeiichi OKUDAMichiaki TERANAKA
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1990 Volume 56 Issue 1 Pages 33-38

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Abstract

Cucumber mosaic virus satellite RNA (CARNA5) was used to investigate the usefulness of replicative form double-stranded RNAs as nonradioactive photobiotin-labelled probes in hybridization studies. In dot-blot hybridization using purified dsCARNA5 labelled with photobiotin as the probe, as little as 2pg of dsCARNA5 was detected. The sensitivity for detecting dsCARNA5 was higher on nylon membranes than on nitrocellulose filters, and dot-blot hybridization was more sensitive than gel electrophoresis with silver staining. CARNA5 in partially purified total nucleic acids extracted from CMV-infected tobacco leaves was readily detected by dot-blot hybridization. Preparations from healthy leaves gave no hybridization signal. In this study three types of CARNA5 were detected and separated by electrophoresis. The three types of CARNA5 hybridized with each other, however, the strongest hybridization signal was obtained with the homologous type of CA-RNA5.

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