Isolation of Cytoplasmic Enzymes from Pollen

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Abstract
The cytoplasmic isozyme of many cytoplasmic-organelle isozyme pairs, as well as other cytoplasmic enzymes in plants, can be readily obtained from pollen [Pisum sativum L. cv. Alaska, Spinacia oleracea L. cv. Early Hybrid No. 7, Zea mays L., Clarkia unguiculata Lindley, Cedrus atlantica Manetti, Oenothera biennis L., Lycopersicon esculentum Mill. and Stephanomeria exigua ssp. coronaria Gottlieb.] by soaking it in an appropriate buffer for 4 h. Enzymes localized in subcellular organelles appear not to be released during the soaking period, although they are released if the pollen is crushed. The technique is a useful initial step in studies of subcellular localization of enzymes or for obtaining small quantities of cytoplasmic enzymes free of organellar contaminants.