Response of Leaf Protein to Ozone in Two White Clover Clones

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Date

1998-05-07

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Publisher

Virginia Tech

Abstract

A white clover ( Trifolium repens ) system, based on one ozone-sensitive (NC-S) and one ozone-resistant (NC-R) clone, has been developed as an indicator to estimate the effects of tropospheric ozone on plant biomass production. A reduction in the vegetative biomass ratio (NC-S/NC-R) of the clones was correlated with increasing concentrations of ozone during a 28-day exposure period. However, the mechanism of ozone sensitivity or tolerance at the biochemical or molecular level is not known. Superoxide dismutase isozyme activities in the two clones did not respond differently to ozone treatment. However, catalase activity increased somewhat more in the leaf tissue of NC-R, compared to NC-S, after ozone treatment. Two-dimensional polyacrylamide gel electrophoresis (PAGE) showed the presence of two proteins (Ozone-Response-Proteins, ORPs) that were more responsive to ozone in the tolerant genotype than in the sensitive one. After ozone treatment for three days, the ORPs were four-fold higher in leaf tissue of NC-R compared to NC-S. Also, the amount of the ORPs was twenty-fold higher in leaf tissue of ozone-treated NC-R than in that of control NC-R. These proteins have apparent molecular weights of 21.5 kD and 23 kD and isoelectric points of 4.1-4.4 on SDS-PAGE gels. The filtrate of a 100 kD concentrator showed that the native molecular weights of the ORPs were less than 100 kD. The results obtained from a study of field samples demonstrated that protein content in leaf tissue of both NC-R and NC-S was positively correlated with ORP content.

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Keywords

White Clover, Ozone-Responsive Protein

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