Date of Award:

5-1971

Document Type:

Thesis

Degree Name:

Master of Science (MS)

Department:

Plants, Soils, and Climate

Department name when degree awarded

Botany

Committee Chair(s)

G. W. Cochran

Committee

G. W. Cochran

Committee

O. S. Cannon

Committee

D. L. Mumford

Committee

W. J. Hanson

Abstract

An organic chemical condensing reagent, 1-cyclohexyl-3-(2-morpholinoethyl)-carbodiimide metho-p-toluene sulfonate (CMC) was tested for effectiveness in inducing polymerization of short lengths of tobacco mosaic virus (TMV) nucleoprotein particles isolated from systemically infected Turkish tobacco leaves. It appeared that the chemical induced the formation of phosphodiester linkages between the viral RNA helices in the joining nucleoprotein fragments. Because the polymerization reaction liberates a hydroxyl ion with each joining event the kinetics of the polymerization can be followed with time using a pH meter. The number of hydroxyl ions liberated is equal to the number of polymerizations. Physical evidence of polymerization was obtained by measuring the lengths of TMV nucleoprotein particles in electron micrographs before and after treating with CMC. The treatments resulted in virus particles that were significantly longer. Biological evidence of polymerization was the fact that the CMC treatment induced significantly greater levels of infectivity in the virus preparations. It appeared that this polymerization phenomenon is essential for the production of infective TMV particles and that the formation of infective TMV units in living cells is probably by the polymerization of shorter units. The TMV nucleoprotein monomer is calculated to be 138 A. U. long while the TMV RNA monomer appears to contain 295 nucleotide units.

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