Detecting Enzyme Variation in the Fern Genus Bommeria: An Analysis of Methodology

Abstract

Numerous flowering plant species were subjected to electrophoretic enzyme analysis and few studies involved pteridophytes. Standard methodology typically results in little or no electrophoretically detectable enzymatic activity in pteridophytes. The presence of large amounts of phenolic compounds in the leaves of most pteridophytes suggests that the electrophoretic difficulties encountered result from complexing of enzymes by these compounds following cellular disruption. The use of a grinding buffer containing dimethylsulfoxide and several compounds inhibitory to phenolic complexing, in combination with a grinding procedure utilizing polyvinylpyrrolidone and liquid N, overcame the difficulties of tissue preparation that result from high concentrations of phenolic compounds. Modifications of this grinding buffer and grinding procedure often resulted in the reduced clarity of enzyme bands, or even a total loss of enzyme activity. In Bommeria, the preparation of leaf tissue in liquid N and use of sodium tetraborate, sodium ascorbate, sodium meta-bisulfite and sodium diethyldithiocarbamate are of less importance than the use of polyvinylpyrrolidone in obtaining active enzyme samples. This procedural data and provision of gel and electrode buffer recipes and staining scheduling should stimulate further electrophoretic investigation of pteridophytes.