Isolation of Sperm Cells from the Pollen of Plumbago zeylanica

Abstract

Intact sperm cells of Plumbago zeylanica were released from mature pollen grains near anthesis using osmotic shock with 20% sucrose. Sperm cell yields of up to 75% can be attained by differential centrifugation using a clinical centrifuge with concentrations of up to 8.8 x 10(6) cells/milliliter. Such ;gametoplasts' remain intact for up to 24 hours according to Evan's blue exclusion and can be used for characterization or physiological manipulation.