During a series of mechanical transfers of tomato spotted wilt virus, two distinct types of mutants were generated. Firstly, a morphologically defective isolate was obtained which had lost the ability to produce the membrane glycoproteins and, as a consequence, was not able to form enveloped particles. Analysis of the genomic RNAs of this isolate suggested that this defect was caused by either point mutations or very small deletions in the medium genomic RNA segment. Secondly, isolates were obtained which had accumulated truncated forms of the large (L) RNA segment. These shortened L RNA molecules most likely represented defective interfering RNAs, since they replicated more rapidly than full-length L RNA and their appearance was often associated with symptom attenuation. Defective L RNAs of different sizes were generated after repeated transfers, and hybridization analysis using L RNA-specific cDNA probes showed that the internal regions deleted varied in length. The presence of defective L RNAs in nucleocapsid fractions as well as in enveloped virus particles indicates that all defective molecules retained the sequences required for replication, encapsidation by nucleocapsid proteins and packaging of the nucleocapsid into virus particles.
de AvilaA. C.,
HuguenotC.,
ResendeR. De. O.,
KitajimaE. W.,
GoldbachR.,
PetersD.1990; Serological differentiation of 20 isolates of tomato spotted wilt virus. Journal of General Virology 71:2801–2807
De HaanP.,
WagemakersL.,
GoldbachR.,
PetersD.1989a; Tomato spotted wilt virus, a new member of the Bunyaviridae?. In Genetics and Pathogenicity of Negative Strand Viruses pp 287–290 Edited by
KolakofskyD.,
MahyB. W. J.
Amsterdam: Elsevier;
De HaanP.,
WagemakersL.,
PetersD.,
GoldbachR.1989b; Molecular cloning and terminal sequence determination of the S and the M RNAs of tomato spotted wilt virus. Journal of General Virology 70:3469–3473
De HaanP.,
WagemakersL.,
PetersD.,
GoldbachR.1990; The S RNA segment of tomato spotted wilt virus has an ambisense character. Journal of General Virology 71:1001–1007
De HaanP.,
KormelinkR.,
ResendeR. de. O.,
van PoelwijkD.,
PetersF.,
GoldbachR.1991; Tomato spotted wilt virus L RNA encodes a putative RNA polymerase. Journal of General Virology 72:2207–2216
De VriesS. C.,
SpringerJ.,
WesselsJ. G. H.1982; Diversity of abundant mRNA sequences and patterns of protein synthesis in etiolated and greened pea seedlings. Planta 156:129–135
FranckiR. I. B.1991; Classification and nomenclature of viruses. Fifth Report of the International Committee of Taxonomy of Viruses.. Archives of Virology (in press)
HuguenotC.,
van den DobbelsteenG.,
de HaanP.,
Wage-makersC. A. M.,
DrostG. A.,
OsterhausA. D. M. E.,
PetersD.1990; Detection of tomato spotted wilt virus using monoclonal antibodies and riboprobes. Archives of Virology 110:47–62
IsmailI. D.,
MilnerJ. J.1988; Isolation of defective interfering particles of sonchus yellow net virus from chronically infected plants. Journal of General Virology 69:999–1006
KnorrD. A.,
MullinR. H.,
HearneP. Q.,
MorrisA. T.1991; De novo generation of defective interfering RNAs of tomato bushy stunt virus by high multiplicity passage. Virology 181:193–202
LiX. H.,
HeatonL. A.,
MorrisT. J.,
SimonA. E.1989; Turnip crinkle virus defective interfering RNAs intensify viral symptoms and are generated de novo. Proceedings of the National Academy of Sciences, U.S.A 86:9173–9177
MorrisT. J.,
KnorrD. A.1990; Defective interfering viruses associated with plant virus infection. In New Aspects of Positive-Strand RNA Viruses pp 123–127 Edited by
BrintonM. A.,
HeinzF. X.
Washington, D. C.: American Society for Microbiology;
NayakD. P.,
ChambersT. M.,
AkkinaR. K.1985; Defective-interfering (DI) RNAs of influenza viruses: origin, structure, expression and interfering. Current Topics in Microbiology and Immunology 114:103–151
NayakD. P.,
ChambersT. M.,
AkkinaR. K.1990; Structure of defective interfering RNAs of influenza viruses and their role in interference. In The Influenza Viruses pp 269–317 Edited by
KrugR. M.
New York: Plenum Press;
ResendeR. de O.,
de AvilaA. C.,
KitajimaE. W.,
GoldbachR.,
PetersD.1991; Defective isolates of tomato spotted wilt virus. In Virus-Thrips-Plant Interactions of Tomato Spotted Wilt Virus, Proceedings of a USDA Workshop, Beltsville, Maryland April 10–19, 1990, pp 71–76 Edited by
HsuH. T.,
LawsonR. H.
United States Department of Agriculture, Agricultural Research Service;
ShirakoY.,
EharaY.1986; Comparison of the in vitro translation products of wild-type and a deletion mutant of soil-borne wheat mosaic virus. Journal of General Virology 67:1237–1245
van LentJ. W. M.,
VerduinB. J. M.1985; Specific gold-labelling of antibodies bound to plant viruses in mixed suspensions. Netherlands Journal of Plant Pathology 91:205–213
VeraniP.,
NicolettiL.,
MarchiA.1984; Establishment and maintenance of persistent infection by the Phlebovirus Toscana in Vero cells. Journal of General Virology 65:367–375