Techniques for clearing ovules for studies of megagametophyte and early postfertilization development in Rhododendron

Biotech Histochem. 1992 Jul;67(4):207-18. doi: 10.3109/10520299209110068.

Abstract

Using ovule clearing, more than 33,600 ovules of Rhododendron nuttallii T. W. Booth (Ericaceae) were examined for megagametophyte and early postfertilization stages at daily intervals from anthesis until 3 weeks after pollination. Pretreatments with amyloglucosidase to digest integumentary and gametophyte starch and Stockwell's bleach to remove tannins from the integumentary epidermis were necessary. Ovules were cleared by a combination or modifications of Herr's four-and-a-half or Stelly's hemalum-methyl salicylate techniques and were observed using differential interference contrast optics. The method proved suitable for large-scale quantitative studies of ovule development and fertilization. A general protocol is suggested as a starting point for ovule clearing studies.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Fertilization
  • Histological Techniques
  • Optics and Photonics
  • Plant Cells*
  • Plant Physiological Phenomena
  • Staining and Labeling / methods