Isolation of Fixed and Viable Eggs, Central Cells, and Embryo Sacs from Ovules of Plumbago zeylanica

Abstract

Three alternative protocols for light microscopy, electron microscopy, and biochemical characterization of isolated megagametophytic tissues are described employing enzymic maceration and microdissection of living and fixed ovular tissue of Plumbago zeylanica. Morphologically well preserved megagametophytes are obtained using fixed ovules in two different regimes (nearly 40 and 60% yield, respectively). Fluorescein diacetate-positive megagametophytic cells are recovered in nearly 20% of unfixed ovules using the third regime.