An immunochemical study of antigen expression in potato spindle tuber viroid (PSTV) - infected tomato leaves and calluses |
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Authors: | J. Matoušek P. Dědič M. J. Beneš P. Kopáček Věra Turková Ludmila Trněná |
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Affiliation: | (1) Institute of Plant Molecular Biology, Czechoslovak Academy of Sciences, Na sádkách 702, 370 05 České Budějovice, Czechoslovakia;(2) Institute of Potato Research and Breeding, Havlíčk∪v Brod, Dobrovského 2366, 58 003 Havlíčk∪v Brod;(3) Institute of Macromolecular Chemistry, CSAS, Heyrovského nám. 2, 162 04 Praha 6;(4) Lab.Anal.Chem. Biological Ctr., CSAS, 370 05 Branišovská, České Budějovice;(5) institute of Experimental Botany, CSAS, Na Perníkářce 59, 160 00 Praha 6, Czechoslovakia |
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Abstract: | A polyspecific antiserum against protein extracted from PSTV-infected tomato leaves was prepared and the IgGs were separated by affinity chromatography on a beaded cellulose adsorbent with an immobilized “healthy” antigen. The antibody not adsorbed entered into a preferential reaction with the antigen from PSTV-infected leaves as estimated by an enzyme-linked immunosorbent assay. The immunochemical reactions did not significantly exceed the control background, if antigens from tomato leaves infected with potato viruses X, Y and M were analyzed. By immunoblot technique we revealed, however, that several antigens not detected in healthy leaves appeared in the leaves infected either with PSTV or with viruses X and M. An accumulation of a major antigen having a molecular mass of about 70 kDa was observed in viroid-infected leaves only, suggesting the specificity for viroid infection. The antigen was found not to be an alkaline endoproteinase - the pathogenesis-related protein P-69. Some antigens with molecular masses approximately 38.0, 23.7 and 22 kDa, which occurred in PSTV-infected leaves and in healthy calluses, were not detectable in PSTV-infected calluses. No reaction exceeding the control level was observed using enzyme-linked immunosorbent assay for antigens from silver nitrate-treated tomato leaves, although such leaves showed symptoms similar to that caused by viroids. |
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