Summary
-
1.
Inocula of TRV produce two types of primary infections: a) Local lesions containing infectious long and non-infectious short virus particles i.e. particulate or “complete” TRV (C-TRV), b) local lesions containing proteinless or “free” infectious RNA of the long particles i.e. “defective” TRV (D-TRV). Both types of lesions are morphologically indistinguishable and their quality needs to be determined in biological tests. The effect of dilution on the occurrence of both types of lesions was studied using artificial mixtures of highly purified long and short particles of a German isolate. The total number of all the lesions produced is directly proportional to the concentration of the infectious long particles present in the inoculum, thus resembling a single hit curve. But, the corresponding curves of the relative numbers of either C-TRV or D-TRV lesions resemble two-hit curves. The percentage of C-TRV lesions decreases by the square of the dilution, whereas the percentage of D-TRV lesions increases in a complementary way.
-
2.
Highly purified long and short particles of TRV were used separately as well as in mixtures for inoculation, and the nature of the resulting local lesions was tested. About 94–100% of all the lesions produced by long particles alone were D-TRV lesions. The short particles did not produce any lesions nor did they multiply detectably. When short particles were added to preparations of homologous long particles neither the morphology nor the absolute number of lesions produced by the latter was affected. But the proportion of C-TRV lesions increased from zero to unity with increasing concentration of the short particles.
-
3.
When non-infectious short particles or their RNA were added to inocula prepared from continuous cultures of homologous “proteinless” D-TRV particulate C-TRV was found to be synthesized in a large number of the resulting local lesions. Such C-TRV cultures as obtained by homologous complementation resembled the parent isolate in all of their properties.
-
4.
Heterologous complementation was carried out using two strains of TRV (TRV-GER and TRV-USA) which are clearly characterized by several genetic markers such as particle size, lesion morphology and coat protein specificity. When the proteinless D-TRV of one strain is complemented with the short particles of the heterologous strain or their RNA, a “mixed” C-TRV is obtained. This “mixed” virus is composed of the long particles resembling the ones of the strain from which the D-TRV originates and of the short particles of the corresponding heterologous strain as used for complementation. The morphology of the local lesions produced by the “mixed” TRV is determined by the RNA of the long particles. Therefore, they resemble the lesions of the C-TRV strain from which the complemented D-TRV originates. Serological tests showed, however, that the “mixed” virus is only distantly related antigenically to this C-TRV isolate. But the “mixed” C-TRV is serologically indistinguishable from the strain from which the complementing short particles were derived.
-
5.
If taken together the four types of evidence presented show, that in the TRV system a full cycle of viral replication is based on the complementing interaction of the RNA of an infectious long and a non-infectious short particle. The infectious “long” RNA appears to be specialized in early functions of viral replication only. It is able to initiate the synthesis of infectious “long” progeny RNA but this RNA remains uncoated by a viral coat protein. The non-infectious “short” RNA, however, appears to be specialized only in late functions and carries the coat protein gene of TRV. For its own replication it is dependent on the replicating “long” RNA. The implications of these findings are discussed.
Similar content being viewed by others
References
Allen jr., T. C.: A strain of tobacco rattle virus from Oregon grown potatoes. The Plant Disease Reporter 47, 920–923 (1963).
Backus, R. C., and R. C. Williams: The use of spraying methods and of volatile suspending media in the preparation of specimens for electron microscopy. J. appl. Phys. 21, 11–15 (1950).
Bawden, F. C.: Speculations on the origins and nature of viruses. In: Plant virology (edit. M. K. Corbett and H. D. Sisler, p. 371. Gainsville: University of Florida Press 1964.
Benzer, S., and S. P. Champe: Ambivalent rH mutants of phage T4. Proc. nat. Acad. Sci. (Wash.) 47, 1025–1038 (1961).
Bockstahler, L. E., and P. Kaesberg: Isolation and properties of RNA from bromegrass mosaic virus. J. molec. Biol. 13, 127–137 (1965).
Boeyé, A., J. L. Melnick, and F. Rapp: SV40-adenovirus hybrids: Presence of two geno-types and the requirement of their complementation for viral replication. Virology 28, 56–70 (1966).
Brandenburg, E., R. Eibner u. R. Tostmann: Untersuchungen über die Eisenfleckigkeit-Pfropfenbildung der Kartoffel als bodengebundene Viruskrankheit. Mitt. Biol. Bundesanst. 97, 36–51 (1959).
Brown, F., S. J. Martin, B. Cartwright, and J. Crick: The ribonucleic acids of the infective and interfering components of vesicular stomatitis virus. J. Gen. Virology 1, 479–486 (1967).
Bruening, G., and H. O. Agrawal: Infectivity of a mixture of cowpea mosaic virus ribonucleoprotein components. Virology 32, 306–320 (1967).
Butel, J. S., and F. Rapp: Replication in simian cells of defective viruses in an SV40-adenovirus “hybrid” population. J. Bact. 91, 278–284 (1966).
Cadman, C. H.: Evidence for association of tobacco rattle virus nucleic acid with a cell component. Nature (Lond.) 193, 49–52 (1962).
—, and B. D. Harrison: Studies on the properties of soil-borne viruses of the tobacco rattle type occuring in Scotland. Anw. appl. Biol. 47, 542–556 (1959).
Davies, P., and R. D. Barry: Nucleic acid of influenza virus. Nature (Lond.) 211, 384 (1966).
Delbrück, M., and W. T. Bailey jr.: Induced mutations in bacterial viruses. Cold. Spr. Harb. Symp. quant. Biol. 11, 33–37 (1946).
Eibner, R.: Untersuchungen über die “Eisenfleckigkeit” der Kartoffel. Thesis Univ. Gießen (1959).
Epstein, R. E., A. Bollé, C. M. Steinberg, E. Kellenberger, E. Boy de la Tour, R. Chevalley, R. S. Edgar, M. Susman, G. Denhardt, and A. Lielausis: Physiological studies on conditional lethal mutations of bacteriophage T4D. Cold Spr. Harb. Symp. quant. Biol. 28, 375–394 (1963).
Frost, R. R., B. D. Harrison, and R. D. Woods: Apparent symbiotic interaction between particles of tobacco rattle virus. J. gen. Virol. 1, 57–69 (1967).
Fulton, R. W.: The effect of dilution on necrotic ringspot virus infectivity and the enhancement of infectivity by noninfective virus. Virology 18, 477–485 (1962).
—: Purification and some properties of tobacco streak and tulare apple mosaic viruses. Virology 32, 153–162 (1967).
Gessner, R.: Zur Frage des Verhaltens und der Übertragung des Tabak-Ratelvirus. Thesis Univ. Gießen (1959).
Gierer, A., and G. Schramm: Infectivity of ribonucleic acid from tobacco mosaic virus. Nature (Lond.) 177, 702–703 (1956).
Hanafusa, H., T. Hanafusa, and H. Rubin: The defectiveness of rous sarcoma virus. Proc. nat. Acad. Sci. (Wash.) 49, 572–580 (1963).
Harrison, B. D., and H. L. Nixon: Separation and properties of particles of tobacco rattle virus with different length. J. Gen. Microbiol. 21, 569–581 (1959a).
——: Some properties of infective preparations made by disrupting tobacco rattle virus with phenol. J. gen. Microbiol. 21, 591 (1959b).
—, and R. D. Woods: Serotypes and particle dimensions of tobacco rattle viruses from Europe and America. Virology 28, 610–620 (1966).
Havranek, P., and J. Zavada: An apparently two-factor dose-response dependence in lucerne mosaic virus assay. Acta virol. 11, 544–550 (1967).
Hirst, G. K.: Genetic recombination with Newcastle disease virus, polioviruses and influenza. Cold Spr. Harb. Symp. quant. Biol. 27, 303 (1962).
Huang, A. S., J. W. Greenawalt, and R. R. Wagner: Defective T-particles of vesicular stomatitis virus 1. Preparation, morphology and biologic properties. Virology 30, 161 (1966).
—, and R. R. Wagner: Defective T-particles of vesicular stomatitis virus. II. Biologic role in homologous interference. Virology 30, 173 (1966).
Huebner, R.: The murine leukemia-sarcoma virus complex. Proc. nat. Acad. Sci. (Wash.) 58, 835–842 (1967).
Kassanis, B.: Properties and behaviour of a virus depending for its multiplication on another. J. gen. Microbiol. 27, 477–488 (1962).
Köhler, E.: Über eine reversible, durch die Jahreszeit induzierte Virulenzänderung beim Tabak-Rattle-Virus. Nachr.bl. dtsch. Pflanzenschutzdienst. Brschw. 8, 93–94 (1956).
—: Die Viren des Kartoffel-Stengelbunt (Tabak-Rattle) und der Pfropfenbildung (Spraing). Eine Stellungnahme zur Frage ihrer Verwandtschaft. Nachr.bl. dtsch. Pflanzenschutzdienst. Brschw. 12, 87–88 (1960).
Lister, R. M.: Possible relationships of virus-specific products of tobacco rattle virus infection. Virology 28, 350–353 (1966).
—: A symptomatological difference between some unstable and stable variants of pea early browning virus. Virology 31, 739–742 (1967).
Novick, A., and L. Szilard: Virus strains of identical phenotype but different genotype. Science 113, 34–35 (1951).
Offord, R. E.: Electron microscopic observation on the substructure of tobacco rattle virus. J. molec. Biol. 17, 370–375 (1966).
—, and J. I. Harris: The protein sub-unit of tobacco rattle virus. Fed. European Biochem. Soc. 2nd Meeting; Abstracts, p. 216–217. Wien: Verlag der Wiener Mediz. Akad. 1965.
Paul, H. L., u. O. Bode: Elektronenmikroskopische Untersuchungen über Kartoffelviren. II. Vermessung der Teilchen von drei Stämmen des Rattle-Virus. Phytopath. Z. 24, 341–351 (1955).
Ralph, R. K., and A. R. Bellamy: Isolation and purification of undegraded ribonucleic acid. Biochim. biophys. Acta (Amst.) 87, 9–16 (1964).
Rapp, F.: Complementation between defective oncogenic viruses. In: Recent results in cancer research VI. Malignant transformation by viruses (edit. W. H. Kirsten, p. 77–94. Berlin-Heidelberg-New York: Springer 1966.
—, J. S. Butel, and J. L. Melnik: SV40-adenovirus “hybrid” populations: Transfer of SV40-determinants from one type of adenovirus to another. Proc. nat. Acad. Sci. (Wash.) 54, 717–724 (1965).
Ravenswaay Claasen, J. C. van: Synthesis of plant viral specific protein in the cell-free system of Escherichia coli. Thesis Univ. of Leiden (1967).
—, A. B. J. van Leeuwen, G. A. H. Duijts, and L. Bosch: In vitro translation of alfalfamosaic virus RNA. J. molec. Biol. 23, 535–544 (1967).
Reichmann, M. E.: The satellite tobacco necrosis virus: A single protein and its genetic code. Proc. nat. Acad. Sci. (Wash.) 52, 1009–1017 (1964).
Rowe, W. P., and S. G. Baum: Studies of adenovirus-SV40 hybrid viruses. II. Defectiveness of the hybrid particles. J. exp. Med. 122, 955–966 (1965).
Rozendaal, A., and J. P. H. van der Want: Over de identiteit van het ratelvirus van de tabak en het stengelbontvirus van de aardappel. T. Plantenziekt. 54, 113–133 (1948).
Sänger, H. L.: Untersuchungen über die beiden charakteristischen Partikel des Ratelvirus und das serologische Verhalten verschiedener Isolate. Thesis Univ. Gießen (1960).
Sänger, H. L.: Untersuchungen über schwer übertragbare Formen des Rattle Virus. Proc. 4th Conf. Potato Virus Diseases, Braunschweig, 22–29 (1960).
—: Defective plant viruses. In: Molecular Genetics, 4. Wiss. Konf. Ges. Dtsch. Naturf. u. Ärzte, Berlin, 1967, Berlin-Heidelberg-New York: Springer 1968.
Sänger, H. L.: Characteristics of tobacco rattle virus. II. Nature and behaviour of defective tobacco rattle virus. In preparation (1968).
—, u. E. Brandenburg: Über die Gewinnung von infektiösem Preßsaft aus “Wintertyp”-Pflanzen des Tabak-Rattle-Virus durch Phenolextraktion. Naturwissenschaften 48, 391 (1960).
Scherr, H. G., A. S. Markowitz, and L. Z. Skelton: A new adjuvant. J. appl. Bact. 28, 174–180 (1965).
Semancik, J. S., and M. R. Kajiyama: Comparative studies on two strains of tobacco rattle virus. J. gen. Virol. 1, 153–162 (1967).
Sol, H. H., and J. W. Seinhorst: The transmission of rattle virus by Trichodorus pachydermus. T. Plantenziekt. 67, 307–309 (1961).
Vooten-Doting, L. van, and E. M. Jaspars: Enhancement of infectivity by combination of two ribonucleic acid components from alfalfa mosaic virus. Virology.
Vloten-Doting, L. van, and E. M. Jaspars: On the infectivity of the RNA's from alfalfa mosaic virus. Fed. of Europ. Biochem. Soc. 4th Meeting, Oslo, Universitätsvorlaget., Abstract-Nr. 446, p. 112 (1967).
Vloten-Doting, L. van, J. Kruseman, and E. M. J. Jaspars: The biological function and mutual dependence of bottom component and top component a of alfalfa mosaic virus. Virology (in the press).
Vogt, P.: A virus released by “nonproducing” rous sarcoma cells. Proc. nat. Acad. Sci. (Wash.) 58, 801–808 (1967).
Watanabe, Y., L. Prevec, and S. F. Graham: Specificity in transcription of the reovirus genome. Proc. nat. Acad. Sci. (Wash.) 58, 1040–1046 (1967).
Wood, H. A., and J. B. Bancroft: Activation of a plant virus by related incomplete nucleoprotein particles. Virology 27, 94–102 (1965).
Yamazaki, H., J. Bancroft, and P. Kaesberg: Biophysical studies of broad bean mottle virus. Proc. nat. Acad. Sci. (Wash.) 47, 979–983 (1961).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Sänger, H.L. Characteristics of tobacco rattle virus. Molec. Gen. Genet. 101, 346–367 (1968). https://doi.org/10.1007/BF00436232
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00436232