A rapid colorimetric method for the determination of tobacco mosaic virus concentration in plant saps

1968 ◽  
Vol 24 (9) ◽  
pp. 972-973
Author(s):  
H. N. Verma ◽  
G. S. Verma
1969 ◽  
Vol 49 (1) ◽  
pp. 112-132
Author(s):  
Justo López García

An investigation of the effects of temperature and of zinc on the development of tobacco-mosaic virus (TMV) in resistant and susceptible varieties of tomatoes was conducted under greenhouse conditions. The study included two experiments, one run during late winter and early spring, and the other during late spring and summer, 1963. Each experiment comprised four treatments and two temperatures. The tomato lines Hawaii 6832, OSU-8, and California 62 PM 22 were used as resistant varieties, whereas the OSU-435-4 line was used as the susceptible one. The common strain of tobacco-mosaic virus was used in this study. Zinc foliar sprays were applied to the tomato plants about 10 days before the seedlings were inoculated with TMV. Immediately after inoculation the plants were divided into two lots and moved to the two temperature houses (cool and hot). The virus concentration was determined by the local lesion assay. One gram of leaf tissue from the tomato plants was ground in a mortar with pestle and a 10-ml. buffer solution added. The extracted sap was squeezed through cheesecloth. One carborundum-dusted leaf of Necrotic Turk tobacco was inoculated. The local lesions were subsequently counted and used as a criterium. Increase in temperature tended to increase virus concentration, although the differences were not significant. Fresh weights of the tomato plants were found to be greater at lower temperatures. Zinc foliar sprays resulted in higher virus concentration and in production of an increased number of local lesions when the inoculum prepared from such treated plants was rubbed into Necrotic Turk tobacco leaves. Effects of zinc on fresh weights were not statistically significant. Zinc applied as foliar sprays appeared to be involved in virus synthesis and multiplication, with higher virus concentrations associated with the addition of zinc. The varieties Hawaii 6832 and OSU-8 were highly resistant, the variety California 62 PM 22 was heterozygous for resistance, and the variety OSU- 435-4 was highly susceptible to the common strain of TMV under the conditions prevailing in these experiments.


1960 ◽  
Vol 15 (1) ◽  
pp. 38-45 ◽  
Author(s):  
Reto Engler ◽  
Gerhard Schramm

Young plants of Nicotiana tabacum (Var. Samsun) were infected with tobacco mosaic virus and kept at a constant temperature between 23 and 27°C and at constant illumination. The virus concentration was determined by bioassay on Nicotiana glutinosa. An exponential increase in virus concentration occurred 20-30 hours after infection. This latent period is significantly shorter after infection with virus ribonucleic acid. Probably the nucleic acid has to be liberated from the nucleoprotein before multiplication can start. The formation and multiplication of free virus ribonucleic acid could be demonstrated earlier than the formation of the complete virus. Infectious nucleic acid was measured by direct extraction of the plants with phenol. Nucleic acid included in the virus was determined after degradation of the free ribonucleic acid by incubation at 37°C and subsequent phenol extraction. The amount of free ribonucleic acid reaches a maximum 40 hours after infection and decreases afterwards to the extent as the virus bound ribonucleic acid increases. A general hypothesis for the biosynthesis of tobacco mosaic virus is given.


1967 ◽  
Vol 35 (1) ◽  
pp. 183-192 ◽  
Author(s):  
M. Weintraub ◽  
H. W. Ragetli ◽  
V. T. John

Tobacco mosaic virus particles were found in small packets and in small numbers, with the electron microscope, in necrotic leaf cells of Nicotiana glutinosa when the samples were fixed in glutaraldehyde and postfixed in OsO4, and the sections were stained with heavy metals. The numbers and size of the virus packets were increased greatly when the leaves were detached from the plant after inoculation Assay of concentration showed that detachment resulted in a 30-fold increase of virus. A similar increase in the number of virus particles detected by electron microscopy was produced by keeping inoculated plants at an air temperature of 26°C. A still greater increase in concentration was effected by incubating detached inoculated leaves at 26°C. Moreover the arrangement of virus particles in these cells resembled that of a systemic virus infection. Cells in local lesions of Chenopodium amaranticolor contained large numbers of virus particles both as packets and in the loose arrangement characteristic of systemic infection. Neither the number of particles nor their arrangement was affected in this host by detaching the leaf or by changing the air temperature. It is suggested that there may be two types of localized virus infections, one of which produces virus in low concentration and is amenable to changes in virus concentration and arrangement as a result of environmental manipulation.


1966 ◽  
Vol 12 (6) ◽  
pp. 369-375 ◽  
Author(s):  
Deana Nobel

Abstract The use of a two-chamber, sealed microdiffusion apparatus has been investigated for the determination of protein nitrogen on samples containing 0.5 mg. protein. The apparatus is constructed from a ground-glass ball-and-socket joint, and all operations including digestion are performed in the designed apparatus. The method has been applied to the determination of nitrogen in tobacco mosaic virus protein and found to be in good agreement with theoretical values.


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