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The Ribonucleic Acid of Tomato Spotted Wilt Virus

Laboratory of Virology, Agricultural University, Binnenhaven 11, Wageningen, The Netherlands

TSWV nucleic acid, extracted from purified virus with phenol-SDS, was non-infectious. Following electrophoresis of the extracted nucleic acid in 2% poly-acrylamide gels containing 0.5% agarose, 3 major and 2 minor bands were observed. The amount of material contained in the different bands varied with the season. Bands 1, 2, 3 and 4 were sensitive to RNases and resistant to DNase. This was a strong indication of the presence of single-stranded RNA in these bands. Precipitation in 2 M-LiCl, effect of heating and mobility in gels containing an increasing percentage of acrylamide confirmed the single-stranded character. Band 1a was resistant to the action of RNases and stained with Coomassie brilliant blue. The mol. wt. of RNAs 1, 2, 3 and 4 (corresponding to bands 1 to 4) were estimated under both non-denaturing and denaturing conditions and appeared to be 2.5 to 2.7 x 10⁶, 1.9 x 10⁶, 1.7 x 10⁶ and 1.3 x 10⁶, respectively. Following dissociation of the virus with Nonidet P 40, an infectious zone could be isolated after sedimentation in an 8 to 43% glycerol gradient.

Received 19 October 1976; accepted 11 February 1977.