Extraction of alcohol dehydrogenase from fresh root tips of loblolly pine

Significant alterations of published methods for extracting alcohol dehydrogenase from plant components were required to obtain measurable enzyme activity from actively growing root tips of loblolly pine (Pinustaeda L.). Necessary steps were quick freezing of roots in liquid nitrogen within 5 min of excising, use of soluble polyvinylpyrrolidone during freezing and fracturing in liquid nitrogen, and use of mercaptoethanol, glycerol, and NAD as protective measures during complete homogenization. No chromatographic purification steps were necessary. Up to 16 samples could be collected and analyzed in a single day by this method with sufficient sensitivity to detect intraspecific variation among loblolly pine families.